Chinese Medicine

Published by BioMed Central
Prescription frequencies of Chinese herbal products (by gender, age and types of herbs), 1997-2003*
Distribution frequencies* of the most commonly prescribed herbal formulae potentially containing aristolochic acid, 1997- 2003
Some Chinese herbal products (CHPs) may contain aristolochic acid (AA) or may be adulterated by the herbs suspected of containing AA which is nephrotoxic and carcinogenic. This study aims to identify the risk and the prescription profile of AA-containing CHPs (AA-CHPs) in Taiwan. A longitudinal analysis was conducted on a randomly sampled cohort of 200,000 patients using the data from the National Health Insurance (NHI) in Taiwan between 1997 and 2003. During the 7-year study period, 78,644 patients were prescribed with AA-CHPs; most patients were females, or middle-aged, or both. A total of 526,867 prescriptions were made to use 1,218 licensed AA-CHPs. Over 85% of the AA-exposed patients took less than 60 g of AA-herbs; however, about 7% were exposed to a cumulated dose of over 100 g of Radix et Rhizoma Asari (Xixin), Caulis Akebiae (Mutong) or Fructus Aristolochiae (Madouling). Patients of respiratory and musculoskeletal diseases received most of the AA-CHP prescriptions. The most frequently prescribed AA-CHPs Shujing Huoxie Tang, Chuanqiong Chadiao San and Longdan Xiegan Tang, containing Radix Stephaniae Tetrandrae, Radix et Rhizoma Asari and Caulis Akebiae, respectively. About one-third of people in Taiwan have been prescribed with AA-CHPs between 1997 and 2003. Although the cumulated doses were not large, further actions should be carried out to ensure the safe use of AA-CHPs.
Frequency distribution of the top 10 disease categories for prescribing CHPs containing phytoestrogens* 
Background Some Chinese herbs contain several kinds of phytoestrogens, and these herbs are commonly prescribed in Taiwan. Phytoestrogens may influence the effects of estrogen in females, although their activities are weak. This study aims to identify the risk and analyze the prescription profile of commonly used phytoestrogenic herbs in Taiwan. Methods The study analyzed women who had been prescribed phytoestrogenic herbs including coumestrol, genistein and/or daidzein between 1997 and 2007 in a fixed cohort taken from all female beneficiaries from the National Health Insurance Research Database of Taiwan. The prescription frequencies, cumulated dosages, and primary indications were listed. Results A total of 462,861 women were included in the study, of whom ~47.0% had used phytoestrogenic herbs at least once during the study period. A total of 6,270,813 prescriptions were recorded, and more than 20% of these contained phytoestrogens. The most commonly prescribed herb and formula were Puerariae Radix and Ge gen tang (Pueraria Decoction), respectively. Most of the prescriptions were issued for diseases of the respiratory system, followed by symptoms, signs, and ill-defined conditions and diseases of the musculoskeletal system and connective tissue. Conclusion This study shows that women who sought medical treatment from Chinese medicine doctors for relief of respiratory discomfort had a high possibility of exposure to phytoestrogenic herbs. Safety issues related to the female endocrine system should be a priority for future research.
TXL's inhibitory effects on cellular growth and apoptosis in HT-29 cells. (A) Apoptosis in HT-29 after TXL treatment for 48 hours was determined by staining the cell with DAPI. Apoptotic cell exhibiting characteristic chromatin condensation were observed by fluorescence microscopy. (B) Cell proliferation was assessed after 24 and 48 hours with the MTT assay as described in Methods. Results (optical densities) are expressed as mean and standard deviation (n = 3). The experiment was repeated three times with similar results. CTL: control.
TXL's effect on the depolarization of HT-29 mitochondria. (A) JC-1 staining observed by fluorescence microscopy. TXL treated cells showed a majority of cells stained green dye due to low mitochondrial membrane potential. (B) Effect of TXL on the depolarization of HT-29 mitochondria was also measured by fluorescence plate reader using JC-1. The cells were exposed to increasing concentrations of TXL. Data represent mean and standard deviation of three individuals with asterisks denoting significant differences between controls and TXL-exposed cells (*P < 0.05, **P < 0.01). CTL: control.
Effect of TXL on cytochrome c. Total protein from HT-29 treated with 1%TXL for 12, 24 and 48 hours (A) or indicated concentration of TXL for 48 hours (B) were analyzed by Western blot with specific antibodies against Bax and Bcl-2. Protein from cytosolic fraction of HT-29 which has been treated with TXL (1%) for 12, 24 and 48 hours (C) or indicated concentration of TXL for 48 hours (D) were analyzed by Western blot with specific antibodies against cytochrome c. GAPDH antibody was used as control for equal loading. The relative expressions of proteins were quantified using Bio-Rad Quatity-One software. Results are expressed as mean and standard deviation (n = 3),*P < 0.05 **P < 0.01compared with control group.
TXL Induces caspase-3 and caspase-9 cleavage. Protein from HT-29 which has been treated with 1%TXL for 12, 24 and 48 hours (A) and protein from HT-29 which has been treated with indicated concentration of TXL for 48 hours (B) were analyzed by Western blot with specific antibodies against caspase-9 and cleaved caspase-3. GAPDH antibody was used as control for equal loading. The relative expressions of proteins were quantified using Bio-Rad Quatity-One software. Results are expressed as means and standard deviation (n = 3),*P < 0.05 **P < 0.01 compared with control group.
Effect of TXL on tumor volume (A) and body weight (B) in HT-29-bearing mouse model. HT-29 cells were injected in nude mice as described in Methods. A total of 24 mice were divided into three groups. Data are expressed as mean and standard deviation (n = 8). Significantly difference compared with control group:*+ P < 0.05, **++ P < 0.01.
Tian Xian Liquid (TXL) is a Chinese medicine decoction and has been used as an anticancer dietary supplement. The present study aims to investigate the effects of TXL on the apoptosis of HT-29 cells and tumor growth in vivo. HT-29 colon cancer cells were treated with gradient dilution of TXL. The mitochondrial membrane potential was measured by JC-1 assay. The release of cytochrome c from mitochondrial and apoptosis-related proteins Bax, Bcl-2, cleaved caspase-3, 9 were examined by Western blot analysis. HT-29 cells were implanted in nude mice to examine the effects of TXL on tumor growth. TXL inhibited HT-29 xenografted model and showed a strong and dose-dependent inhibitory effect on the proliferation of HT-29 cells. Mitochondrial membrane potential was reduced by TXL at the concentration of 0.5% above. For Western blot analysis, an increase in Bax expression and a decrease in Bcl-2 expression were observed in TXL-treated cells. TXL treatment increased the protein level of cleaved casepase-3 and caspase-9, and the release of cytochrome c in cytoplasm was up-regulated as well. TXL significantly inhibits cell proliferation in the HT-29 cells and HT-29 xenografted model via the mitochondrial cell death pathway.
Transcriptional activation of ApoA-IV mRNA by lipid micelles. A: Caco-2/TC7 cells were treated with lipid micelles (1:1000 and 1:2500) for 24 hours. Total RNAs were extracted and reverse transcribed to cDNA for real-time PCR analysis. The mRNA levels of ApoA-IV were determined by the Ct-value method and normalized by mRNA level of a house keeping gene 18S rRNA. B: Caco-2/TC7 cells were treated with various concentrations of lipid micelles for 48 hours. Cultures were collected for luciferase assay to determine ApoA-IV transcription. C: Caco-2/TC7 cells were treated with lipid micelles (1:2500 and 1:2000) for 24, 48 and 72 hours to measure the time-dependent regulation of ApoA-IV. Data are expressed as mean ± SD of the multiple of Basal (i.e. buffer-treated control set as one) and number of independent experiments (n) = 5.
Transcriptional activation of ApoA-IV by Chinese medicinal herbs. Caco-2/TC7 cells were treated in various groups with water (1 mg/ml) or ethanol (0.1 mg/ml) extracts of the Chinese medicinal herbs for 48 hours. Luciferase activity was measured. Left: A: Rhizoma Alismatis; B: Fructus Crataegi; C: Semen Coicis;D: Rhizoma Atractylodis Macroczphalae; E: Rhizoma Atractylodis; F: Sclerotium Poriae Cocos; G: Semen Cassiae; H: Folium Sennae); Right: I: Radix Angelica Sinensis; J: Rhizoma Curcumae; K: Flos Chrysanthemi; L: Radix et Rhizoma Notoginseng; M: Folium Nelumbinis; N: Herba Taraxaci; O: Pericarpium Citri Retiiculatae; P: Fructus Schisandrae; Q: Frutus Mori. Data are expressed as mean ± SD of the multiple fold of control (i.e. buffer-treated control set as one) and number of independent experiments (n) = 5; P < 0.05 (*); P < 0.01 (**).
Stimulation of ApoA-IV mRNA by Rhizoma Alismatis and Radix Angelica Sinensis. A: Caco-2/TC7 cells were treated with water extracts (0–10 mg/ml) of Rhizoma Alismatis and Radix Angelica Sinensis for 48 hours. Luciferase activity was measured. B: Caco-2/TC7 cells were treated with Rhizoma Alismatis and Radix Angelica Sinensis (1 mg/ml) for 24 hours. The change of ApoA-IV mRNA was determined by RT-PCR analysis. Data are expressed as mean ± SD of the multiple of Basal (i.e. water-treated control set as one) and number of independent experiments (n) = 5.
Reduction of TG content by Radix Angelica Sinensis and Rhizoma Alismatis in differentiated 3T3-L1 adipocytes. A: Lipogenic differentiated 3T3-L1 cells were either serum-starved or treated with insulin (10 μg/ml) for three days, and then stained by Oil red O dye. The amount of stained TG (red color) was quantified at 490 nm absorbance. B: Lipogenic differentiated 3T3-L1 cells were treated with Rhizoma Alismatis and Radix Angelica Sinensis (0.1, 1 and 10 mg/ml) for 72 hours. The reduction of TG content was measured. Data are expressed as mean ± SD of the percentage of control (i.e. water-treated control set as 100) and number of independent experiments (n) = 5; P < 0.01 (**); P < 0.001 (***).
Chinese medicine has been proposed as a novel strategy for the prevention of metabolic disorders such as obesity. The present study tested 17 Chinese medicinal herbs were tested for their potential anti-obesity effects. The herbs were evaluated in terms of their abilities to stimulate the transcription of Apolipoprotein A-IV (ApoA-IV) in cultured Caco-2/TC7 enterocytes. The herbs that showed stimulating effects on ApoA-IV transcription were further evaluated in terms of their abilities to reduce the formation of triglyceride in differentiated 3T3-L1 adipocytes. ApoA-IV transcription was stimulated by Rhizoma Alismatis and Radix Angelica Sinensis in a dose- and time-dependent manner in cultured Caco-2/TC7 cells. Moreover, these two herbs reduced the amount of triglyceride in differentiated 3T3-L1 adipocytes. The results suggest that Rhizoma Alistmatis and Radix Angelica Sinensis may have potential anti-obesity effects as they stimulate ApoA-IV transcription and reduce triglyceride formation.
Chrysin (5,7-dihydroxyflavone) inhibits platelet-derived growth factor-induced vascular smooth muscle cell proliferation and arterial intima hyperplasia. This study aims to investigate the effects of chrysin on rat pulmonary vascular remodeling in hypoxia-induced pulmonary hypertension (PH). Sprague-Dawley rats were continuously exposed to 10% O2 for 4 weeks to induce PH. The effect of chrysin (50 or 100 mg/kg/d, subcutaneous) on vascular remodeling was investigated in hypoxia-induced PH model. At the end of the experiments, the indexes for pulmonary vascular remodeling and right ventricle hypertrophy were measured by vascular medial wall thickness and the ratio of right ventricle to (left ventricle plus septum). The expressions of NOX4, collagen I, and collagen III were analyzed by immunohistochemistry, real-time PCR, or western blotting. The proliferation of cultured pulmonary artery smooth muscle cells (PASMCs) was determined by BrdU incorporation and flow cytometry. The levels of malondialdehyde (MDA) and reactive oxygen species (ROS) were also determined by thiobarbituric acid reactive substances assay and 2'7'-dichlorofluorescein diacetate method. Chrysin treatment for 4 weeks significantly attenuated pulmonary vascular remodeling and improved collagen accumulation and down-regulated collagen I and collagen III expressions, accompanied by downregulation of NOX4 expression in the pulmonary artery (P = 0.012 for 50 mg/kg/d, P < 0.001 for 100 mg/kg/d) and lung tissue (P = 0.026, P < 0.001). In vitro, chrysin (1, 10, and 100 μM) remarkably attenuated PASMC proliferation (P = 0.021 for 1 μM, P < 0.001 for 10 μM, and P < 0.001 for 100 μM), collagen I expression (P = 0.035, P < 0.001, and P < 0.001), and collagen III expression (P = 0.027, P < 0.001, and P < 0.001) induced by hypoxia, and these inhibitory effects of chrysin were accompanied by inhibition of NOX4 expression (P = 0.019, P < 0.001, and P < 0.001), ROS production (P = 0.038, P < 0.001, and P < 0.001), and MDA generation (P = 0.024, P < 0.001, and P < 0.001). This study demonstrated that chrysin treatment in hypoxia-induced PH in rats reversed the hypoxia-induced (1) elevations of NOX4 expression, (2) productions of ROS and MDA, (3) proliferation of PASMC, and (4) accumulation of collagen.
Polysaccharopeptide (PSP) from Coriolus versicolor (Yunzhi) is used as a supplementary cancer treatment in Asia. The present study aims to investigate whether PSP pre-treatment can increase the response of the human leukemia HL-60 cells to apoptosis induction by Camptothecin (CPT). We used bivariate bromodeoxyuridine/propidium iodide (BrdUrd/PI) flow cytometry analysis to measure the relative movement (RM) of the BrdUrd positively labeled cells and DNA synthesis time (Ts) on the HL-60 cell line. We used annexin V/PI flow cytometry analysis to quantify the viable, necrotic and apoptotic cells. The expression of cyclin E and cyclin B1 was determined with annexin V/PI flow cytometry and western blotting. Human peripheral blood mononuclear cells were used to test the cytotoxicity of PSP and CPT. PSP reduced cellular proliferation; inhibited cells progression through both S and G2 phase, reduced 3H-thymidine uptake and prolonged DNA synthesis time (Ts) in HL-60 cells. PSP-pretreated cells enhanced the cytotoxicity of CPT. The sensitivity of cells to the cytotoxic effects of CPT was seen to be the highest in the S-phase and to a small extent of the G2 phase of the cell cycle. On the other hand, no cell death (measured by annexin V/PI) was evident with the normal human peripheral blood mononuclear cells with treatment of either PSP or CPT. The present study shows that PSP increases the sensitization of the HL-60 cells to undergo effective apoptotic cell death induced by CPT. The pattern of sensitivity of cancer cells is similar to that of HL-60 cells. PSP rapidly arrests and/or kills cells in S-phase and did not interfere with the anticancer action of CPT. PSP is a potential adjuvant to treat human leukemia as rapidly proliferating tumors is characterized by a high proportion of S-phase cells.
Infantile colic is a common painful clinical condition associated with signs of distended intestines and an increase in colon peristalsis. However, clinical documentation of observed gastrointestinal functions in the condition is still lacking. Even though the ailment is common, no clear treatment guidelines exist. While acupuncture with minimal stimulation has been shown to be effective in reducing crying behaviour of infants suffering from colic, the documented effect of acupuncture on gastrointestinal function in children with infantile colic is scarce. This case series study aims to document the symptoms of routinely rated gastrointestinal function and the changes in these symptoms after minimal acupuncture in a larger group of children with infantile colic. This study included 913 infants with normal weights, and lengths at birth. The infants' mean age was 5.4 weeks when the observations started, and had colic symptoms since two weeks after birth. Light needling stimulation of the acupuncture point LI4 was performed for 10-20 seconds bilaterally on a daily basis for a mean of 6.2 consecutive days. A questionnaire with verbal rating scales for the parents' evaluation was used before and after the treatment period. Before treatment the infants were assessed by the parents in terms of 'often have inflated stomachs' (99%) and 'seldom drool' (76%), 'regurgitate' (53%) and 'belch' (62%). Moreover, the reported frequency of defecation was 5-8 times per day (64%), with a yellowish-green colour (61%) and with a water-thin consistency (74%). After treatment, the variables of inflated stomachs, drooling and regurgitating were systematically changed, and rated by the parents as occurring 'sometimes' while belching was rated as occurring 'often' and the frequency of defecation was reduced to 1-4 times/day with a mustard yellow colour and a gruel-like consistency. The parents also rated their impression of the infants' general colic symptoms including crying behaviour as much ameliorated in 76% of the cases. The results of the present study show that minimal acupuncture at LI4 in infantile colic is an effective and easy treatment procedure that, furthermore, is reported to be without serious side effects.
Isodeoxyelephantopin (IDOE) isolated from Elephantopus scaber L. (Didancao) is used in Chinese medicine for the treatment of some types of cancer. The anti-cancer mechanism of IDOE remains unclear. This study aims to investigate the antiproliferative activity of IDOE on breast carcinoma T47D cells and lung carcinoma A549 cells. The growth inhibitory effects of IDOE on breast carcinoma T47D cells, lung carcinoma A549 cells, and normal lymphocytes were evaluated by the MTT assay. Morphological analysis of apoptosis induction was performed by acridine orange/ethidium bromide dual-staining and Hoechst 33342 nuclear staining. The cell cycle profile, caspase-3 expression, and annexin V staining were evaluated by flow cytometry. IDOE inhibited the growth of A549 and T47D cells in a dose- and time-dependent manner with IC50 values of 10.46 and 1.3 mug/mL, respectively. IDOE was not significantly toxic to normal lymphocytes. The cells became detached from the monolayer and rounded up, had fragmented nuclei and condensed chromatin, and the numbers of apoptotic cells increased (P = 0.0003). IDOE-induced cell death was associated with activated caspase-3 expression followed by cell cycle arrest at G2/M phase. IDOE inhibited the proliferation of breast cancer cells and lung carcinoma cells and induced caspase-3-mediated apoptosis and cell cycle arrest in the treated cells.
Effects of COP and BER on the growth of A549 cells. A. Morphological changes of A549 cells treated with 6.4 μg/mL COP or 4.0 μg/mL BER for 72 h under inverted phase contrast microscope (200 ×). B. Treatment with COP and BER for 72 h inhibited A549 cell growth (P < 0.01) [Pearson Correlation Test, P = 0.0032 for COP; P = 0.0178 for BER] in a dose-dependent manner. Values are means ± SD of 4 independent assays.
Growth inhibition by combined use of COP/BER and chemotherapeutic agents in A549 cells. Effects of COP or BER, combined with 5-fluorouracil (5-FU), camptothecin (CPT), or paclitaxel (TAX), on the growth of A549 cells. COP-L: 1.6 μg/mL; COP-H: 6.4 μg/mL; BER-L: 0.5 μg/mL; BER-H: 4 μg/mL. *P < 0.05, compared to drugs alone. Values are means ± SD of 4 independent assays.
Effects of COP and BER on ROS production in A549 cells. Cells were treated with COP-L (1.6 μg/mL), COP-H (6.4 μg/mL), BER-L (0.5 μg/mL), or BER-H (4 μg/mL) for 24 h and harvested for ROS determination. * P < 0.05 compared to control. Values are means ± SD of 4 independent assays.
Effects of COP and BER on MDR in A549 cells. Cells were treated with COP-L (1.6 μg/mL), COP-H (6.4 μg/mL), BER-L (0.5 μg/mL), or BER-H (4 μg/mL) for 24 h and harvested for dye retention determination. * P < 0.05 compared to control. Values are means ± SD of 4 independent assays.
Non-small cell lung cancer is associated with high expression of multidrug resistance (MDR) proteins and low production of reactive oxygen species (ROS). Coptis extract (COP), a Chinese medicinal herb, and its major constituent, berberine (BER), have anticancer properties. This study aims to investigate the effects of COP and BER combined with chemotherapeutic agents, including fluorouracil (5-FU), camptothecin (CPT), and paclitaxel (TAX), on cell proliferation, ROS production, and MDR in A549 human non-small cell lung cancer cells. A549 cells were treated with different doses of COP and BER, combined with 5-FU, CPT, and TAX. Cell viability was measured by an XTT (2,3-bis-(2-methoxy-4- nitro-5-sulfophenyl)-2 H-tetrazolium-5-carboxanilide) assay. Intracellular ROS levels were determined by measuring the oxidative conversion of cell permeable 2',7'-dichlorofluorescein diacetate to fluorescent dichlorofluorescein. MDR of A549 cells was assessed by rhodamine 123 retention assay. Both COP and BER significantly inhibited A549 cell growth in a dose-dependent manner. Combinations of COP or BER with chemotherapeutic agents (5-FU, CPT, and TAX) exhibited a stronger inhibitory effect on A549 cell growth. In addition, COP and BER increased ROS production and reduced MDR in A549 cells. As potential adjuvants to chemotherapy for non-small cell lung cancer, COP and BER increase ROS production, reduce MDR, and enhance the inhibitory effects of chemotherapeutic agents on A549 cell growth.
Bi-directional transport of the selected isoflavones in Caco-2 monolayer model
Chemical structures of the three selected isoflavones. A Daidzin; B Daidzein; C Puerarin.
Basolateral-to-apical transport of daidzin in the absence and presence of MK 571, the MRP inhibitors.
The HPLC-UV chromatograms of daidzin before and after incubations with rat intestinal microvilli preparations. I.S.: internal standard.
The HPLC-UV chromatograms of puerarin before and after incubations with rat intestinal microvilli preparations. I.S.: internal standard.
Radix Puerariae (Gegen) contains abundant isoflavones in the forms of glycosides and aglycones, such as daidzein, daidzin and puerarin. This study aims to investigate the intestinal absorbability and mechanism of these three structurally related isoflavones. The bi-directional transport of these three isoflavones in Caco-2 monolayer model was performed to evaluate their absorbability and involvement of transporters in Transwell. In vitro incubation of daidzin and puerarin with rat intestinal microvilli preparation was conducted to estimate their potential form of absorption in vivo. Daidzein demonstrated passive diffusion transport while puerarin did not. Daidzin showed basolateral-to-apical transport and the absorption extent could be reduced by 50% in the presence of MK571, a multidrug resistance-associated protein inhibitor (MRP). The in vitro incubation study of daidzin and puerarin indicated that daidzin was hydrolyzed to daidzein whereas puerarin remained unchanged. While daidzein was transported more efficiently, puerarin was resistant to intestinal hydrolysis and inefficiently transported across intestinal epithelium. Daidzin demonstrated a low intestinal absorbability due to a significant efflux transport mediated by MRPs. Daidzin was likely to be hydrolyzed by intestinal microvilli and subsequently released daidzein for intestinal absorption.
HPLC chromatograms of the aerial part of Z. Clinopodioides . (A) Acacetin. (B) Extract of sample.
Accuracy of the HPLC method for the determination of acacetin
Effects of different acacetin concentrations on cardiomyocyte viability
The total flavonoids from ethanol extract of the aerial part of Ziziphora clinopodioides Lam. (Lamlaceae) (Xintahua) showed protective activities against rat acute myocardial ischemia in rats. This study aims to isolate acacetin, a flavonoid, from the aerial part of Z. clinopodioides, to develop an HPLC method for its detection, and to evaluate its protective effects on neonatal rat cardiomyocytes. Sephadex LH-20 silicagel and pillar layer chromatography silica gel were applied for the isolation and purification of acacetin and its structure was elucidated on the basis of (1)H and (13)C NMR spectroscopy. The content of acacetin in Z. clinopodioides collected from three different origins was determined by HPLC. The neonatal rat cardiomyocytes were isolated and cultured in vitro to establish a hypoxia/reoxygenation injury model. The viability of cardiomyocytes was measured by the MTT method. Changes of malondialdehyde (MDA) content in the medium were also determined. The acacetin content in various batches of Z. clinopodioides ranged from 45.50 to 47.41 μg/g. Acacetin of 25, 10, 5 μg/mL significantly decreased the MDA content in a model of hypoxia/reoxygenation injury (P < 0.001, P < 0.001 and P = 0.033, respectively). Acacetin protects neonatal cardiomyocytes from the damage induced by hypoxia/reoxygenation stress through reduction of lipid peroxidation and enhancement of the antioxidant activity.
Chotosan (CTS, Diaoteng San), a Kampo medicine (ie Chinese medicine) formula, is reportedly effective in the treatment of patients with cerebral ischemic insults. This study aims to evaluate the therapeutic potential of CTS in cognitive deficits and investigates the effects and molecular mechanism(s) of CTS on learning and memory deficits and emotional abnormality in an animal aging model, namely 20-week-old senescence-accelerated prone mice (SAMP8), with and without a transient ischemic insult (T2VO). Age-matched senescence-resistant inbred strain mice (SAMR1) were used as control. SAMP8 received T2VO (T2VO-SAMP8) or sham operation (sham-SAMP8) at day 0. These SAMP8 groups were administered CTS (750 mg/kg, p.o.) or water daily for three weeks from day 3. Compared with the control group, both sham-SAMP8 and T2VO-SAMP8 groups exhibited cognitive deficits in the object discrimination and water maze tests and emotional abnormality in the elevated plus maze test. T2VO significantly exacerbated spatial cognitive deficits of SAMP8 elucidated by the water maze test. CTS administration ameliorated the cognitive deficits and emotional abnormality of sham- and T2VO-SAMP8 groups. Western blotting and immunohistochemical studies revealed a marked decrease in the levels of phosphorylated forms of neuroplasticity-related proteins, N-methyl-D-aspartate receptor 1 (NMDAR1), Ca2+/calmodulin-dependent protein kinase II (CaMKII), cyclic AMP responsive element binding protein (CREB) and brain-derived neurotrophic factor (BDNF) in the frontal cortices of sham-SAMP8 and T2VO-SAMP8. Moreover, these animal groups showed significantly reduced levels of vasculogenesis/angiogenesis factors, vascular endothelial growth factor (VEGF), VEGF receptor type 2 (VEGFR2), platelet-derived growth factor-A (PDGF-A) and PDGF receptor α (PDGFRα). CTS treatment reversed the expression levels of these factors down-regulated in the brains of sham- and T2VO-SAMP8. Recovery of impaired neuroplasticity system and VEGF/PDGF systems may play a role in the ameliorative effects of CTS on cognitive dysfunction caused by aging and ischemic insult.
Recovery rates of five main saponins in P. notogingseng
HPLC profiles of the seed (A), six-month-old root (B), adult root (C) and seed MeOH extract (D) of Panax notoginseng.
Time courses of saponin accumulation in Panax notoginseng seed and seedling root. The seeds were collected in November and stored in wet sand. They began to germinate the following January. The seedlings were then allowed to grow for one year in soil nursery bed.
Panax notoginseng is an important Chinese medicinal plant. Dammarene-type triterpenoid saponins are main pharmacologically effective compounds in P. notoginseng. This study aims to investigate the formation and accumulation of saponins in P. notoginseng roots during germination and juvenile stage. P. notoginseng seeds were collected and stored in wet sand. After germination, the seedlings were transplanted into a soil nursery bed and cultivated for one year. During this period, samples were collected every month and the concentrations of ginsengnosides Rg1, Re, Rb1, Rd and notoginsengnoside R1 were determined by HPLC. There was little saponin in the P. notoginseng seed. The chemical composition of seed was different from that of root. After germination, Rb1, Rg1, Re, Rd and R1 appeared successively in the seedling root. And in the five-month-old root, all these five main saponins came into existence. The accumulation of saponins in P. notoginseng root was affected by seasons. The accumulation of saponins showed a time-dependent increase after germination of P. notoginseng.
Flowchart describing the simulation study. Departing from Zang-fu single patterns dataset, manifestation profiles were simulated according to the combination of traditional examination methods. Cases (true positive) and controls (true negative) profiles were tested with criteria F% and N% and with F% and N%-cutoff for comparison of diagnostic tests. PD+: successful pattern differentiation; PD-: unsuccessful pattern differentiation.
Percent time interval for execution of the simulation (MPSA) and identification (PDA) algorithms. The combination of methods progressively increased the duration of both simulation and identification processes.
Receiver operating characteristic curves with respect to the available information for diagnosis. The combination of methods yielded distinct cutoff points. The Four Examinations (Ip+AO+Iq+P) resulted in the best overall statistical performance with the minimum cutoff value of available information, followed by three (Ip+AO+Iq), two (Ip+AO) and single (Ip) examination methods.
Average normalized accuracy as a function of the available information for diagnosis. All tested combinations provided concave-shaped curves, indicating that there is an optimum quantity of available information to accurately perform pattern differentiation. Also, inspection (Ip) or its combination with auscultation-olfaction (Ip+AO) provided lower average normalized accuracies than addition of inquiry (Ip+AO+Iq) and palpation (Ip+AO+Iq +P; Four Examinations) to the examination, as indicated by the relative position of the concavity (maximum value).
Average number of candidate patterns as a function of available information for diagnosis. The average normalized number of candidate patterns increased as more information was available to perform pattern differentiation.
Clinical practice of Chinese medicine requires little information for differentiation of Zang-fu patterns. This study is to test the impact of information amount on the diagnostic accuracy of pattern differentiation algorithm (PDA) using stochastic simulation of cases. A dataset with 69 Zang-fu single patterns was used with manifestations according to the Four Examinations, namely inspection (Ip), auscultation and olfaction (AO), inquiry (Iq) and palpation (P). A variable quantity of available information (N(%)) was randomly sampled to generate 100 true positive and 100 true negative manifestation profiles per pattern to which PDA was applied. Four runs of simulations were used according to the Four Examinations: Ip, Ip+AO, Ip+AO+Iq and Ip+AO+Iq+P. The algorithm performed pattern differentiation by ranking a list of diagnostic hypotheses by the amount of explained information F(%). Accuracy, sensitivity, specificity and negative and positive predictive values were calculated. Use the Four Examinations resulted in the best accuracy with the smallest cutoff value (N(%)= 28.5%), followed by Ip+AO+Iq (33.5%), Ip+AO (51.5%) and Ip (52.0%). All tested combinations provided concave-shaped curves for accuracy, indicating an optimal value subject to N(%-cutoff). Use of N(%-cutoff) as a secondary criterion resulted in 94.7% (94.3; 95.1) accuracy, 89.8% (89.1; 90.6) sensitivity, and 99.5% (99.3; 99.7) specificity with the Four Examinations. Pattern differentiation based on both explained and optimum available information (F(%) and N(%-cutoff)) is more accurate than using explained and available information without cutoff (F(%) and N(%)). Both F(%) and N(%-cutoff) should be used as PDA's objective criteria to perform Zang-fu single pattern differentiation.
Hemodynamic characteristics of rat hearts subjected to ischemia reperfusion
Free radical scavenging activities of Desmodium gangeticum extract
Effects of ethyl acetate root extract of Desmodium gangeticum on TBARS, catalase, superoxide dismutase (SOD), and glutathione peroxidase (GPx) in the tissue homogenate of isolated rat hearts
Activities of creatine kinase, lactate dehydrogenase, SGOT, and SGPT in the tissue homogenate of isolated rat hearts
Activities of creatine kinase and lactate dehydrogenase in the perfusate of isolated rat hearts. Group 1: normal control; Group 2.1, 2.2, 2.3: ischemic reperfusion control; Group 3.1, 3.2, 3.3: drug pretreated and subjected to ischemic reperfusion. Values are mean ± SD in each group (n = 6).
This study aims to evaluate the antioxidant potential of the ethyl acetate extract of Desmodium gangeticum root for cardioprotection from ischemia reperfusion-induced oxidative stress. The in vitro antioxidant potential of the extract was in terms of hydroxyl radical scavenging activity, lipid peroxide scavenging activity, nitric oxide scavenging activity and diphenylpicrylhydrazyl radical scavenging activity. The in vivo antioxidant potential of the extract was assessed in an isolated rat heart model. Free radicals were scavenged by the extract in a concentration-dependent manner within the range of the given concentrations in all models. Administration of the ethyl acetate extract of Desmodium gangeticum root (100 mg per kg body weight) before global ischemia caused a significant improvement of cardiac function and a decrease in the release of lactate dehydrogenase in coronary effluent, as well as the level of malondialdehyde in myocardial tissues. The ethyl acetate extract of Desmodium gangeticum root protects the myocardium against ischemia-reperfusion-induced damage in rats. The effects of the extract may be related to the inhibition of lipid peroxidation.
Shikonin derivatives have cytotoxic and antitumor effects. This study aims to investigate the antitumor effects of acetylshikonin isolated from a Chinese medicinal herb Arnebia euchroma (Royle) Johnst. The 3-(4,5-dimethylthiazol- 2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to determine the in vitro antitumor effects of acetylshikonin on human lung adenocarcinoma cell line A549, human hepatocellular carcinoma cell line Bel-7402, human breast adenocarcinoma cell line MCF-7 and mouse Lewis lung carcinoma (LLC) cell line. C57BL/6 mice with LLC model were used to study the in vivo antitumor effects of acetylshikonin. The expression of bax, bcl-2 and caspase-3 proteins in LLC tissue was determined with immunohistochemical staining. In A549, Bel-7402, MCF-7 and LLC cell lines, acetylshikonin inhibited cell growth in a dose-dependent manner. IC50 (means +/- SD) were 5.6 +/- 0.86 microg/ml, 6.82 +/- 1.5 microg/ml, 3.04 +/- 0.44 microg/ml and 2.72 +/- 0.38 microg/ml respectively. Acetylshikonin suppressed tumor growth in C57BL/6 mice with LLC. The inhibition rate of acetylshikonin (2 mg/kg) was 42.85%. Immunohistochemical staining revealed that in the acetylshikonin groups the expression of bax and caspase-3 increased, whereas the expression of bcl-2 decreased, suggesting that acetylshikonin induced tumor cell apoptosis through activating the pro-apoptotic bcl-2 family and caspase-3. Acetylshikonin isolated from Arnebia euchroma (Royle) Johnst cell suspension cultures exhibits specific in vivo and in vitro antitumor effects.
Background The land and ecology suitability for producing pollution-free Chinese medicinal materials should be evaluated based on Geographic Information System (GIS). This study aims to determine production regions for pollution-free Achyranthes bidentata in Tianjin as a case to illustrate the workflow based on GIS. Methods The slopes, land usage, residential areas and roads were selected to evaluate the land suitability, to avoid the potential pollution sources. The ecology suitability evaluation was performed based on the soil type and nine climate factors, such as active accumulated temperature, mean January temperature, mean July temperature, the lowest temperature in January, the highest temperature in July, mean annual temperature, sunshine duration, relative humidity, annual precipitation, affecting the natural growth of A. bidentata. Results The best production regions for pollution-free A. bidentata in Tianjin, with a total area of approximately 575 km2, were found in Jinghai County, Ninghe County, Wuqing District, and Dagang District. Conclusion This study illustrated a workflow based on GIS for determining the production regions in Tianjin for pollution-free A. bidentata.
ABSTRACT: Interstitial pulmonary fibrosis is characterized by an altered cellular composition of the alveolar region with excessive deposition of collagen. Lung inflammation is also common in pulmonary fibrosis. This study aims to test the inhibition of 5-lipooxygenase (5-LOX) by boswellic acid (BA) extract in an experimental model of pulmonary fibrosis using bleomycin (BL). Boswellic acid extract (1 g/kg) was force-fed to rats seven days prior to administration of BL or gamma irradiation or both. BL (0.15 U/rat) in 25 μl of 0.9% normal saline (NS) or 0.9% NS alone was administered intratracheally. Rats were exposed to two fractionated doses of gamma irradiation (0.5 Gy/dose/week) with a gamma cell-40 (Cesium-137 irradiation units, Canada) during the last two weeks of the experiment. BA was administered during BL or irradiation treatment or both. After the animals were sacrificed, bronchoalveolar lavage was performed; lungs were weighed and processed separately for biochemical and histological studies. In rats treated with BL, levels of transforming growth factor-β1 (TGF-β1) and tumor necrosis factor-α (TNF-α) were significantly elevated (P = 0.05 and P = 0.005). Hydroxyproline was highly and extensively expressed. Immunoreactive compounds were abundantly expressed, represented in the levels of macrophages infiltrate, accumulation of eosinophils and neutrophils in the lung as well as the aggregation of fibroblasts in the fibrotic area. The levels of lipoxygenase enzyme activity were significantly increased (P = 0.005). Antioxidant activities measured in BL-treated rats deteriorated, coupled with the elevation of both levels of plasma lipid peroxide (LP) content and bronchoalveolar lavage lactate dehydrogenase activity. BA-treated rats had reduced number of macrophages, (P = 0.01), neutrophils in bronchoalveolar lavage (P = 0.01) and protein (P = 0.0001). Moreover, the hydroxyproline content was significantly lowered in BA-treated rats (P = 0.005). BA extract inhibited the TGF-ß induced fibrosis (P = 0.01) and 5-LOX activity levels (P = 0.005).Histologically, BA reduced the number of infiltrating cells, ameliorated the destruction of lung architecture and attenuated lung fibrosis. BA attenuates the BL-induced injury response in rats, such as collagen accumulation, airway dysfunction and injury. This study suggests that the blocking of 5-LOX may prevent the progression of fibrosis.
Benzoylmesaconine (BMA) is the main Aconitum alkaloid in Radix Aconiti Lateralis Preparata (Fuzi, aconite roots) with potent pharmacological activities, such as analgesia and anti-inflammation. The present study developed a simple and reliable method using BMA as a marker compound for the quality control of processed aconite roots and their products. After extraction, a high-performance liquid chromatography (HPLC) determination of BMA was conducted on a RP-C18 column by gradient elution with acetonitrile and aqueous phase, containing 0.1% phosphoric acid adjusted with triethylamine to pH 3.0. A distinct peak profile was obtained and separation of BMA was achieved. Method validation showed that the relative standard deviations (RSDs) of the precision of BMA in all intra-day and inter-day assays were less than 1.36%, and that the average recovery rate was 96.95%. Quantitative analysis of BMA showed that the content of BMA varied significantly in processed aconite roots and their products. This HPLC method using BMA as a marker compound is applicable to the quality control of processed aconite roots and their products.
Asaronaldehyde (2, 4, 5-trimethoxybeznaldehyde) is an active component of Acorus gramineus rhizome. This study aims to evaluate the anti-Candida efficacy of asaronaldehyde and its three structural isomers, namely, 2, 3, 4-trimethoxybenzaldehyde, 3, 4, 5-trimethoxybenzaldehyde, and 2, 4, 6- trimethoxybenzaldehyde. Susceptibility testing of test compounds was carried out using standard methodology (M27-A2) as per clinical and laboratory standards institute guidelines. Minimum fungicidal concentration (MFC) was determined as the lowest concentration of drug killing 99.9% of Candida cells. The effect on sterol profile was evaluated using the ergosterol quantitation method. Effects on morphogenesis, adhesion and biofilm formation in C. albicans were studied using germ-tube, adherence and biofilm formation assays respectively. Cytotoxicity of test compounds to human RBCs was determined by hemolysis assay. 2, 4, 6-Trimethoxybenzaldehyde exhibited significant anti-Candida activity (P = 0.0412). Minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) were established as 0.25 and 0.5 mg/mL, respectively. All of the test compounds showed significant inhibition of hyphal form transition in yeast at MIC/2 and MIC/4 values. 3, 4, 5-Trimethoxybenzaldehyde and 2, 4, 6-trimethoxybenzaldehyde inhibited adhesion and biofilms. A hemolytic assay of these compounds revealed that they were non-toxic at MIC values. Asaronaldehyde reduced sterol content. Asaronaldehyde and 2, 4, 6-trimethoxybenzaldehyde showed anti-Candida efficacy.
Herbs commonly used in diabetes management
Insulin secretion and pancreatic-β-cell apoptosis. Glucose is taken up into β-cells via glucose transporters. It is metabolized in glycolysis and Krebs cycle, resulting in an increased ratio of ATP to ADP in the cytoplasm. This closes ATP-sensitive potassium channels (KATP channels), leading to cell membrane depolarization and subsequently opening voltage-gated Ca2+ channels. These changes increase free Ca2+ concentration ([Ca2+]i) in cytoplasm and eventually triggers insulin secretion. In apoptosis, stimuli promotes the release of caspase activators from mitochondria and result in the activation of caspases procedure, by cleaving the effector caspases, which interacts with a variety of cellular proteins, resulting in directly or indirectly the morphological and biochemical characteristics of cell apoptosis. The action sites of hypoglycemia herbs are indicated with a narrow.
Insulin signal pathway and insulin insensitive. The inner part of IR reveals a tyrosine kinase activity and coupled with proteins of Src-homology-collagen-like protein (SHC) and multifunctional docking proteins IRS-1 and IRS-2. The interaction of insulin and IR activates its tyrosine activity and phosphorylates the coupled SHC and subsequently activates, in turn, a series of signal proteins, including the growth factor receptor-binding protein 2 (Grb2), and the ras small guanosine 5'-triphosphate-binding protein. The in turn signaling leads to an activation of the MAPK cascade involved in mitogenesis and the open status of a hexose transporter protein (GLUTs), which is located in the cell membrane and is the only pump to take into glucose for cells. The decreased serine/threonine phosphorylation of IR, inactivates hexokinase and glycogen synthase, as well as defects in the phosphorylation of glucose transporter protein (GLUT4) and genetic primary defect in mitochondrial fatty acid oxidation, leading to insulin resistance and an increase of triglyceride synthesis contribute to this insulin insensitivity. The action sites of hypoglycemia herbs are indicated with an arrow.
Action sites of western medicine in diabetes treatment. Hypoglycemic medicines restore euglycemia via several types, including insulin secretagogues (sulfonylureas, meglitinides), insulin sensitizers (biguanides, metformin, thiazolidinediones), alpha-glucosidase inhibitors (miglitol, acarbose).
Action sites of herbs in diabetes treatment. The efficacy of hypoglycemia herbs has been mediated by increasing insulin secretion (ginseng, bitter melon, aloes, biophytum sensitivum), enhancing glucose uptake by adipose and muscle tissues (ginseng, bitter melon and cinnamon), inhibiting glucose absorption from intestine (myrcia and sanzhi) and inhibiting glucose production from heptocytes (berberine, fenurgreek leaves).
Conventional drugs treat diabetes by improving insulin sensitivity, increasing insulin production and/or decreasing the amount of glucose in blood. Several herbal preparations are used to treat diabetes, but their reported hypoglycemic effects are complex or even paradoxical in some cases. This article reviews recent findings about some of the most popular hypoglycemic herbs, such as ginseng, bitter melon and Coptis chinensis. Several popular commercially available herbal preparations are also discussed, including ADHF (anti-diabetes herbal formulation), Jiangtangkeli, YGD (Yerbe Mate-Guarana-Damiana) and BN (Byakko-ka-ninjin-to). The efficacy of hypoglycemic herbs is achieved by increasing insulin secretion, enhancing glucose uptake by adipose and muscle tissues, inhibiting glucose absorption from intestine and inhibiting glucose production from heptocytes.
Liver fibrosis is a common histological process to develop into cirrhosis in various chronic liver diseases including chronic hepatitis and fatty liver. Therefore anti-liver fibrosis is very important strategy to treat chronic liver diseases. Fuzheng Huayu (FZHY), a preparation containing herbs such as Radix Salvia Miltiorrhizae, Cordyceps, Semen Persicae, was formulated on the basis of Chinese medicine theory in treating liver fibrosis and was approved. Pharmacological studies and clinical trials demonstrate that FZHY has a significant effect against liver fibrosis and that many of the pharmacological actions are attributable to the effect. This article reviews the effects and actions of FZHY, in particular the effects observed from clinical trials in treating liver fibrosis caused by chronic hepatitis B and the actions on inhibition of hepatic stellate cell activation, protection of hepatocytes and inhibition of hepatic sinusoidal capillarization. This article also reviews the coordinated effects of the constituent herbs of FZHY and the actions of their active compounds such as salvianonic acid B (SA-B) on liver fibrosis.
Carcinogenesis is a multiple steps process. The initiation step is started by the transformation of the normal cell into a cancer cell (initiated cell). These cells undergo tumour promotion into preneoplastic cells, which progress to neoplastic cells. Inflammation and oxidative stress, together with the accumulation of genetic alterations over a lifetime of patients, will result in the formation of cancer. It is important to take note that in reality, cancer may arise without proceeding through each of these steps. Chemopreventive agents can interfere with different steps of this process. Some agents inhibit metabolic activation of the procarcinogens to their ultimate electrophilic species, or their subsequent interaction with DNA. These agents therefore block tumour initiation (blocking agents). Alternatively, blocking agents can stimulate the detoxification of carcinogens, leading to their excretion from the body. Other agents suppress (suppressing agents) the later steps (promotion and progression). Some agents can act as both blocking and suppressing agents.
Schematic presentation of Nrf2-ARE pathway. In the cytoplasm, under basal level, newly synthesized Nrf2 is constitutively bound to Keap1 forming a dimer, Nrf2-Keap1. Keap1 is a cytosolic protein that inhibits Nrf2 signalling by promoting Nrf2 degradation through proteasomal pathway. When oxidants such as ROS, RNS and dietary chemopreventive compounds react with redox reactive cysteines in Keap1, Nrf2 will be released from Keap1, hence allowing the transcriptional factor Nrf2 to translocate to the nucleus. In the nucleus, Nrf2 dimerizes with basic leucine zipper partners (bZip) such as small MAF-family proteins and bind to ARE, which is located in the promoter of the phase II and antioxidative genes, triggering the transcription of ARE-regulated genes. The critical role of Nrf2 in protecting cells/subjects from neoplastic transformation when subject to oxidative stress and carcinogens is performed by enhancing the expression of detoxifying metabolizing enzymes and maintaining oxidative stress homeostasis by producing antioxidant enzymes. Application of chemopreventive compounds can further enhance the expression of phase II detoxifying and antioxidant enzymes by up-regulating the Nrf2-ARE expression.
A simplified illustration showing the role of Nrf2 in anti-oxidative and anti-inflammatory pathways preventing carcinogenesis. Upon stimulation by ROS, RNS (having negative effects in subjects) and chemopreventive compounds (having positive effects in subjects), Nrf2 is activated and NF-kB pathways can also be mediated concurrently, such multiple interactions allow chemopreventive compounds, including ginseng, to exert their beneficial cancer preventive and therapeutic effects.
This article reviews recent basic and clinical studies of ginseng, particularly the anti-cancer effects and the potential chemopreventive actions by activating the transcriptional factor, nuclear factor (erythroid-derived 2)-like 2 (Nrf2 or NFE2L2)-mediated anti-oxidative stress or anti-inflammatory pathways. Nrf2 is a novel target for cancer prevention as it regulates the antioxidant responsive element (ARE), a critical regulatory element in the promoter region of genes encoding cellular phase II detoxifying and anti-oxidative stress enzymes. The studies on the chemopreventive effects of ginseng or its components/products showed that Nrf2 could also be a target for ginseng's actions. A number of papers also demonstrated the anti-inflammatory effects of ginseng. Targeting Nrf2 pathway is a novel approach to the investigation of ginseng's cancer chemopreventive actions, including some oxidative stress and inflammatory conditions responsible for the initiation, promotion and progression of carcinogenesis.
In Chinese medicine, ginseng (Panax ginseng C.A. Meyer) has long been used as a general tonic or an adaptogen to promote longevity and enhance bodily functions. It has also been claimed to be effective in combating stress, fatigue, oxidants, cancer and diabetes mellitus. Most of the pharmacological actions of ginseng are attributed to one type of its constituents, namely the ginsenosides. In this review, we focus on the recent advances in the study of ginsenosides on angiogenesis which is related to many pathological conditions including tumor progression and cardiovascular dysfunctions. Angiogenesis in the human body is regulated by two sets of counteracting factors, angiogenic stimulators and inhibitors. The 'Yin and Yang' action of ginseng on angiomodulation was paralleled by the experimental data showing angiogenesis was indeed related to the compositional ratio between ginsenosides Rg1 and Rb1. Rg1 was later found to stimulate angiogenesis through augmenting the production of nitric oxide (NO) and vascular endothelial growth factor (VEGF). Mechanistic studies revealed that such responses were mediated through the PI3K→Akt pathway. By means of DNA microarray, a group of genes related to cell adhesion, migration and cytoskeleton were found to be up-regulated in endothelial cells. These gene products may interact in a hierarchical cascade pattern to modulate cell architectural dynamics which is concomitant to the observed phenomena in angiogenesis. By contrast, the anti-tumor and anti-angiogenic effects of ginsenosides (e.g. Rg3 and Rh2) have been demonstrated in various models of tumor and endothelial cells, indicating that ginsenosides with opposing activities are present in ginseng. Ginsenosides and Panax ginseng extracts have been shown to exert protective effects on vascular dysfunctions, such as hypertension, atherosclerotic disorders and ischemic injury. Recent work has demonstrates the target molecules of ginsenosides to be a group of nuclear steroid hormone receptors. These lines of evidence support that the interaction between ginsenosides and various nuclear steroid hormone receptors may explain the diverse pharmacological activities of ginseng. These findings may also lead to development of more efficacious ginseng-derived therapeutics for angiogenesis-related diseases.
Immuno-modulatory effects of ginseng, including both immuno-stimulatory and immuno-suppressive effects, have been widely reported. This study aims to determine whether the paradoxical immuno-modulatory effect is related to unique phytochemical profiles of different North American (NA) ginseng, namely aqueous (AQ) and alcoholic (ALC) extracts. AQ and ALC extracts were prepared and their immuno-bioactivity were studied in vitro in murine macrophages (Raw 264.7) through measuring the direct stimulatory production of pro-inflammatory mediator and cytokines as well as the suppression of lipopolysaccharide (LPS)-stimulatory response by the two extracts. Gel permeation chromatography was used to fractionate and isolate phytochemicals for characterization of ginseng extracts. AQ extract up-regulated the production of nitric oxide (NO), tumour necrosis factor-α (TNF-α) and interleukin-6 (IL-6) while ALC extract did not. ALC extract but not AQ extract suppressed LPS-induced macrophage NO and TNF-α production. These immuno-stimulatory and suppressive effects were exhibited at similar extract concentrations. Moreover, the macrophage-stimulating activity of the AQ extract was inhibited in the presence of ALC extract. Fractionation of AQ extract revealed the presence of two major peaks at 230 nm with average molecular weights of 73,000 and 37,000 Da. The first fraction had similar elution volume as the crude polysaccharide (PS) fraction isolated from the AQ extract, and it was the only bioactive species. Parallel fractionation study of ALC extract yielded similar elution profiles; however, both sub-fractions were devoid of PS. Fraction I of the ALC extract suppressed LPS-induced NO production dose-dependently. ALC extract of NA ginseng, which was devoid of PS, was immuno-inhibitory whereas the AQ extract, which contained PS, was immuno-stimulatory. These extract-related anti-inflammatory and pro-inflammatory effects may be considered as the Yin and Yang actions of ginseng.
Electroacupuncture (EA) has been used to treat inflammatory diseases. Alternatively activated macrophages (AAMo) stimulated by cytokines such as interleukin (IL)-4, IL-10 and IL-13 are anti-inflammatory and mildly microbicidal. This study aimed to evaluate whether EA at the Zusanli acupoint (ST36) would change the profile of healthy murine macrophages, particularly the generation of AAMo and susceptibility to Leishmania major infection. BALB/c mice were treated with EA (15/30 Hz) at the ST36 acupoint for 20 min/d for 5 d. After the final EA session, the mice were euthanized and their peritoneal cells were harvested and counted for determination of arginase activity, nitric oxide (NO) production and microbicidal activity after culture in the presence or absence of IL-4, interferon-γ (IFNγ) or lipopolysaccharide (LPS) or both IFNγ and LPS. Twelve mice were infected with L. major promastigotes into the footpads after the final EA session and the infection course was monitored. Peritoneal cells freshly obtained from EA-treated mice had similar arginase and microbicidal activities to cells from sham-treated mice. After culture with IL-4, cells from EA-treated mice exhibited significant increases in the arginase activity (sham: 58 ± 11.3 vs. EA: 80.7 ± 4.6%, P = 0.025) and number of parasites/infected cell (sham: 2.5 ± 0.4 vs. EA: 4.3 ± 0.8 cells, P = 0.007). The NO production was lower in cells from EA-treated mice cultured in the presence of a combination of IFNγ and LPS (sham: 31.6 ± 6.5 vs. EA: 22.3 ± 2.1 μM, P = 0.025). The lesion size in mice infected with L. major promastigotes was larger in EA-treated mice (sham: 3.26 ± 0.29 vs. EA: 2.23 ± 0.4 mm, P = 0.039). EA at the ST36 acupoint increases IL-4 responsiveness in macrophages, Generation of AAMo and susceptibility to L. major infection.
Harmine is a beta-carboline alkaloid from the plant Peganum harmala. Previous studies found that harmine inhibited metastasis of B16F-10 melanoma cells. This study aims to elucidate the role of harmine in apoptosis of B16F-10 cells. B16F-10 melanoma cells were treated in the presence and absence of harmine in vitro. Morphological changes, cell cycle and expression of various pro and anti- apoptotic genes were analyzed for the study of apoptosis. Morphological observation and DNA laddering assay showed that harmine treated cells displayed marked apoptotic characteristics, such as nuclear fragmentation, appearance of apoptotic bodies and DNA laddering fragment. TUNEL assay and flow cytometric analysis also confirmed apoptosis. Furthermore, RT-PCR analysis showed that harmine induced apoptosis in B16F-10 melanoma cells by up-regulating Bax and activating Caspase-3, 9 and p53 and down-regulating Bcl-2. Harmine also up-regulated Caspase-8 and Bid, indicating that harmine affected both extrinsic and intrinsic pathways of apoptosis. This study also showed inhibitory effects of harmine on some transcription factors and pro- inflammatory cytokines that protect cell from apoptosis. Harmine activates both intrinsic and extrinsic pathways of apoptosis and regulates some transcription factors and pro-inflammatory cytokines.
The role of Chinese herbs and its active ingredients on proliferation and differentiation of neural stem cells in vitro 
The role of Chinese herbs and its active ingredients to promote bone marrow mesenchymal stem cells differentiate into nerve cells 
Some Chinese herbs are anti-thrombolysis, and anti-inflammatory, improves brain RNA content, promotes brain protein synthesis, enhances dopamine function, regulates brain hormones, and improves microcirculation in central nervous system that might improve, repair and rehabilitation from the stroke and brain injury. Specific Chinese herbs and their components, such as Acanthopanax, Angelica, could maintain the survival of neural stem cells, and Rhodiola, Ganoderma spore Polygala, Tetramethylpyrazine, Gardenia, Astragaloside and Ginsenoside Rg1 promoted proliferation of neural stem cells, and Rhodiola, Astragaloside promoted differentiation of neural stem cell into neuron and glia in vivo. Astragalus, Safflower, Musk, Baicalin, Geniposide, Ginkgolide B, Cili polysaccharide, Salidroside, Astragaloside, Antler polypeptides, Ginsenoside Rg1, Panax notoginseng saponins promoted proliferation and differentiation of neural stem cells in vitro. Salvia, Astragalus, Ginsenoside Rg1, P. notoginseng saponins, Musk polypeptide, Muscone and Ginkgolide B promoted neural-directed differentiation of MSCs into nerve cells. These findings are encouraging further research into the Chinese herbs for developing drugs in treating patients of stroke and brain injury.
Nasopharyngeal carcinoma (NPC) has a high incidence rate in Southern China. Although there are conventional therapies, the side effects and toxicities are not always tolerable for patients. Recently, the tumoricidal effect of ginsenosides on different cancer cells has been studied. This study aims to investigate the anti-cancer effect of ginsenosides on NPC cells and their underlying mechanism. The cytotoxicity of ginsenosides on NPC cell line HK-1 was measured by MTT assay. Apoptosis was detected by propidium iodide staining followed by flow cytometry. A xenograft tumor model was established by injecting nude mice with HK-1 cells. The activation of caspases and apoptosis-inducing factor (AIF) were evaluated by Western blot analysis. Nuclear translocation of AIF was also studied by immunofluorescence staining. Mitochondrial membrane potential was measured by JC-1 dye using flow cytometry. Four ginsenosides, 20 (S)-Rh2, compound K (CK), panaxadiol (PD) and protopanaxadiol (PPD), induced apoptotic cell death in HK-1 cells in a concentration-dependent manner. CK inhibited HK-1 xenograft tumor growth most extensively and depleted mitochondrial membrane potential depolarization and induced translocation of AIF from cytoplasm to nucleus in HK-1 cells. In addition, depletion of AIF by siRNA abolished CK-induced HK-1 cell death. Ginsenoside CK-induced apoptosis of HK-1 cells was mediated by the mitochondrial pathway and could significantly inhibit tumor growth in vivo.
Antifibrotic medicinal plants targeting HSC activation and ECM deposition. HSC: hepatic stellate cells, ECM: extracellular matrix, 1: C. longsa, S. marianum, G. biloba, S. miltiorrhiza, G. glabra, S. baicalensis, B. falcatum, Phyllanthus species, B. aristata, Ginseng species, A. paniculata, and Coffea species. 2: C. longa, S. marianum, G. biloba, S. miltiorrhiza, G. glabra, S. baicalensis, B. falcatum, Phyllanthus species, B. aristata, Ginseng species, and Coffea species.
Inhibitory mechanisms of hepatic stellate cell activation via medicinal plants
Inhibitory mechanisms of extracellular matrix deposition via medicinal plants
During chronic liver injury, hepatic stellate cells (HSC) are activated and proliferate, which causes excessive extracellular matrix (ECM) deposition, leading to scar formation and fibrosis. Medicinal plants are gaining popularity as antifibrotic agents, and are often safe, cost-effective, and versatile. This review aims to describe the protective role and mechanisms of medicinal plants in the inhibition of HSC activation and ECM deposition during the pathogenesis of liver fibrosis. A systematic literature review on the anti-fibrotic mechanisms of hepatoprotective plants was performed in PubMed, which yielded articles about twelve relevant plants. Many of these plants act via disruption of the transforming growth factor beta 1 signaling pathway, possibly through reduction in oxidative stress. This reduction could explain the inhibition of HSC activation and reduction in ECM deposition. Medicinal plants could be a source of anti-liver fibrosis compounds.
Ethnomedicinal uses and properties of the selected plants
Solid content of extracts obtained by different methods*. *Error bars represent standard deviation (n = 3).
Antioxidant activities of extracts tested by DPPH assay*. *Error bars represent standard deviations (n = 3).
Eight medicinal plants were tested for their antimicrobial and antioxidant activities. Different extraction methods were also tested for their effects on the bioactivities of the medicinal plants. Eight plants, namely Herba Polygonis Hydropiperis (Laliaocao), Folium Murraya Koenigii (Jialiye), Rhizoma Arachis Hypogea (Huashenggen), Herba Houttuyniae (Yuxingcao), Epipremnum pinnatum (Pashulong), Rhizoma Typhonium Flagelliforme (Laoshuyu), Cortex Magnoliae Officinalis (Houpo) and Rhizoma Imperatae (Baimaogen) were investigated for their potential antimicrobial and antioxidant properties. Extracts of Cortex Magnoliae Officinalis had the strongest activities against M. Smegmatis, C. albicans, B. subtilis and S. aureus. Boiled extracts of Cortex Magnoliae Officinalis, Folium Murraya Koenigii, Herba Polygonis Hydropiperis and Herba Houttuyniae demonstrated greater antioxidant activities than other tested medicinal plants. Among the eight tested medicinal plants, Cortex Magnoliae Officinalis showed the highest antimicrobial and antioxidant activities. Different methods of extraction yield different spectra of bioactivities.
The active components of Panax quinquefolius activate the cytochrome-c related Bcl-2 and inhibit UCP-2. The activation of cytochrome-c related Bcl-2 has negative effects on caspase-9 triggered apoptotic cascade and then apoptosis. As UCP-2 induces apoptotic cascade and decreases ATP production, the inhibition of UCP-2 has negative effects on apoptosis and positive effects on ATP production, thereby increasing insulin production and release.  : up-regulated.  : down-regulated. + : positive effects. - : negative effects.  : known actions.  : recently discovered actions.
The mechanism of the beneficial effects of Panax quinquefolius (Xiyangshen, American ginseng) on diabetes is yet to be elucidated. Recent studies show that Panax quinquefolius increases insulin production and reduces the death of pancreatic beta cells. Mechanism studies indicate that Panax quinquefolius improves cell's immuno-reactivity and mitochondrial function through various factors. Clinical studies show that Panax quinquefolius improves postprandial glycemia in type 2 diabetic patients. Further studies to identify the component(s) of Panax quinquefolius linked with pancreatic islets/beta cells in vitro and in vivo are warranted for better understanding of the full effects of Panax quinquefolius.
Background This study aims to determine the relationship between the antioxidant and anti-inflammatory activities of the thirteen herbs and two fungi extracts, and their total phenolic and flavonoid contents. Methods Antioxidant activities were evaluated by four assays: an antioxidant activity assay using Saccharomyces cerevisiae, a DPPH ((2, 2-diphenyl-1-picrylhydrazyl) assay to assess free radical scavenging, an assay assessing ferrous ions or iron (II) chelating ability, and a ferric reducing antioxidant power (FRAP) assay. Total phenolic and flavonoid contents were determined using the Folin-Ciocalteu and aluminium chloride methods, respectively. Anti-inflammatory activities were determined by measuring the inhibition of nitric oxide and TNF-α production in lipopolysaccharide- and interferon-γ-activated J774A.1 macrophages. Their cytotoxicities against macrophages were determined by MTT assay. Results A positive linear correlation between antioxidant activities and the total phenolic and flavonoid content of the plant extracts was found. The plant extracts with high phenolic and flavonoid content also exhibited significant anti-inflammatory activity with good cell viability. Conclusion The selected herbs could be a rich source of antioxidants and free radical scavenging compounds. The levels of phenolic and flavonoid compounds were correlated with the antioxidant and anti-inflammatory activities of the extracts from the herbs.
Structures of the four isomers in silymarin.
Metabolism of glycyrrhizin catalyzed by β-D-glucuronidase in human liver and intestines.
Structures of the dibenzocyclooctadiene compounds and osthole.
Structures of the saikosaponin and ginsenoside compounds.
Damaging effects of oxyradical on biological systems and sites of blockades () by antioxidants/free radical scavengers (adapted and expanded from reference [61]).
A literature survey was conducted on herbs, their preparations and ingredients with reported liver protection activities, in which a total of 274 different species and hundreds of active ingredients have been examined. These ingredients can be roughly classified into two categories according to their activities: (1) the main ingredients, such as silybin, osthole, coumarin, glycyrrhizin, saikosaponin A, schisandrin A, flavonoids; and (2) supporting substances, such as sugars, amino acids, resins, tannins and volatile oil. Among them, some active ingredients have hepatoprotective activities (e.g. anti-inflammatory, anticancer, antioxidant, immunomodulating and liver cirrhosis-regulating effects). Calculation of physicochemical parameters indicates that the main ingredients with negative and positive Elumo values possibly display their hepatoprotective effects through different mechanisms, such as antioxidative, anti-inflammatory and immunomodulating effects. As the combination of herbs may achieve some treatment effects synergistically and/or additively, it is common in Chinese medicine to use mixtures of various medicinal herbs with pharmacologically active compounds to have synergistic and/or additive effects, or to reduce harmful effects of some pharmacologically active compounds. In particular, the active compounds with Clog P around 2 are suitable for passive transport across membranes and accessible to the target sites. Thus, Elumo and Clog P values are good indicators among the calculated parameters. Seven different physicochemical parameters (MW, Clog P, CMR, μ, Ehomo, Elumo and Hf) and four major biological activities (antioxidant, anti-inflammatory, antiviral/antitumor and immunomodulating) are discussed in this review. It is hoped that the discussion may provide some leads in the development of new hepatoprotective drugs.
This study aims to isolate the alpha-glucosidase inhibitory compounds from mulberry leaves (Morus atropurpurea Roxb., Moraceae) and to develop an analytical method for quantification of the compounds. Four flavonoids, rutin (1), isoquercetin (2), kaempferol-3-O-rutinoside (3) and astragalin (4), were isolated by column chromatography from mulberry leaf water extracts (MWE). The alpha-glucosidase inhibitory activities of MWE and the four isolated compounds were evaluated by a microplate-based in vitro assay. The content of the isolated flavonoids in M. atropurpurea leaves purchased from different local herbal stores or collected in different locations was determined by high performance liquid chromatography. The four flavonoids (1--4) showed alpha-glucosidase inhibitory activities, with rutin (1) and astragalin (4) showing high alpha-glucosidase inhibitory activities (IC50 values of 13.19 +/- 1.10 and 15.82 +/- 1.11 muM, respectively). The total contents of the four flavonoids were different among eight samples examined, ranging from 4.34 mg/g to 0.53 mg/g. The four flavonoids in M. atropurpurea leaves could inhibit alpha-glucosidase activity.
Histopathologic sections of theliver in the CCl4-induced model. (Group I) Liver sections of normal healthy mice show normal arrangements of cells in the liver lobules. (Group II) Hepatocyte necrosis and evident vacuolation of hepatocytes are observed in liver sections of CCl4-treated mice. (Group III and Group IV) Liver sections of mice pretreated with bifendate at 0.40 g/kg body weight (Group III) or SRF at 0.05 g/kg body weight (Group IV) show mild vacuolation of hepatocytes. (Group V) Mice pretreated with SRF (0.10 mg/kg body weight) show very mild hepatocyte necrosis and mild vacuolation of hepatocytes. (Group VI) Liver sections of mice pretreated with SRF (0.20 mg/kg body weight) show marked improvement in the histology compared with the CCl4-treated control group. HE staining; 100× magnification.
Histopathologic sections of theliver in the BCG+ LPS-induced model. (Group I) Liver sections of normal healthy mice show normal arrangements of cells in the liver lobules. (Group II) Hepatocyte necrosis and evident vacuolation of hepatocytes are observed in liver sections of BCG + LPS-treated mice. (Group III and Group IV) Liver sections of mice pretreated with bifendate at 0.40 g/kg body weight (Group III) or SRF at 0.05 g/kg body weight (Group IV) show mild vacuolation of hepatocytes. (Group V and Group VI) Mice pretreated with SRF at 0.10 mg/kg body weight (Group V) or 0.20 mg/kg body weight (Group VI) show marked improvement in the histology compared with the CCl4-treated control group. HE staining; 100× magnification.
Background Cichorium glandulosum Boiss. et Huet is used for treatment of liver disorders, and its effects are attributed to sesquiterpenes. This study aims to investigate the hepatoprotective effects of a sesquiterpene-rich fraction (SRF) from the aerial part of C. glandulosum on carbon tetrachloride (CCl4)-induced acute hepatotoxicity in mice, and on priming with Bacillus Calmette–Guerin (BCG) followed by lipopolysaccharide (LPS)-induced immunological liver injury in mice. Methods SRF was suspended in water and administered to mice at 0.05, 0.10 and 0.20 g/kg body weight for 7 consecutive days. An active control drug (bifendate pills) was suspended in distilled water and administered to mice at 0.40 g/kg body weight for 7 consecutive days. Hepatotoxicity was induced by intraperitoneal injection of 0.1% CCl4 (0.2 mL/mouse) at 13 h before the last drug administration, or by tail intravenous injection of BCG (0.2 mL/mouse) before the first drug administration and LPS (0.2 mL/mouse; 8 μg) at 15 h before the last drug administration. Blood samples and the livers were collected for evaluation of the biochemical parameters of aspartate aminotransferase (AST), alanine aminotransferase (ALT) and total bilirubin (TBIL). Results SRF significantly reduced the impact of CCl4 toxicity. The highest dose of SRF (0.20 g/kg) was the most effective, reflected by significant reductions in the levels of AST (P = 0.001), ALT (P = 0.000) and TBIL (P = 0.009). The serum enzymatic levels induced by BCG and subsequent LPS injection were significantly and dose-dependently restored by SRF, reflected by significant reductions in the levels of AST (P = 0.003), ALT (P = 0.003) and TBIL (P = 0.007) for the highest dose of SRF (0.20 g/kg). Conclusion SRF is hepatoprotective in animal models of chemical and immunological acute liver injury.
Morphological appearance of 
Antrodia cinnamomea
Antrodia cinnamomea (AC) is an endemic mushroom species of Taiwan, and has been demonstrated to possess diverse biological and pharmacological activities, such as anti-hypertension, anti-hyperlipidemia, anti-inflammation, anti-oxidation, anti-tumor, and immunomodulation. This review focuses on the inhibitory effects of AC on hepatitis, hepatocarcinoma, and alcohol-induced liver diseases (e.g., fatty liver, fibrosis). The relevant biochemical and molecular mechanisms are addressed. Overall, this review summarizes the hepatoprotective activities in vitro and in vivo. However, there is no doubt that human and clinical trials are still limited, and further studies are required for the development of AC-related products.
Nomenclature and classification of selected Yang and Yin tonifying herbs * Pharmaceutical name (Chinese pinyin) Plant part used Latin botanical name
Anti-aging potential of Chinese tonifying herbs.
Chinese tonifying herbs such as herba cistanche, ganoderma and cordyceps, which possess antioxidant and/or immunomodulatory activities, can be useful in the prevention and treatment of age-related diseases. Pharmacological studies on Yang and Yin tonifying herbs suggest that Yang tonifying herbs stimulate mitochondrial adenosine triphosphate (ATP) generation, presumably through the intermediacy of reactive oxidant species, leading to the enhancement of cellular/mitochondrial antioxidant status. Yin tonifying herbs, however, apart from possessing antioxidant properties, exert mainly immunomodulatory functions that may boost a weak immune system and may also suppress overreactive immune responses. The abilities of Yang and Yin Chinese tonifying herbs to enhance ATP generation and to exhibit antioxidant and/or immunomodulatory actions are the pharmacological basis for their beneficial effects on the retardation of aging.
Transcutaneous electric acupoint stimulation (TEAS) avoids the use of needles, and instead delivers a mild electric current at traditional acupoints. This technique has been used for treating heroin addiction, but has not been systematically tested for other drugs of abuse. This study aims to investigate the effects of TEAS on drug addiction. Volunteers who were either cocaine-dependent (n = 9) or cannabis-dependent (n = 11) but were not seeking treatment for their dependence participated in a within-subject, single-blind study. Treatment consisted of twice daily 30-minute sessions of TEAS or sham stimulation for 3.5 days. The active TEAS levels were individually adjusted to produce a distinct twitching response in the fingers, while the sham stimulation involved 2 minutes of stimulation at threshold levels followed by 28 minutes of stimulation below the detection levels. The participants recorded their drug use and drug cravings daily. At 1 hour after the last morning session of TEAS or sham stimulation, a cue-induced craving EEG evaluation was conducted. Event-related P300 potentials (ERPs) were recorded, sorted, and analyzed for specific image types (neutral objects, non-drug-related arousing images, or drug-related images). TEAS treatment did not significantly reduce the drug use or drug cravings, or significantly alter the ERP peak voltage or latency to peak response. However, the TEAS treatment did significantly modulate several self-reported measures of mood and anxiety. The results of this pilot study with a limited sample size suggest that the acupoint stimulation techniques and protocol used in this trial alone do not significantly reduce cravings for or use of cocaine or cannabis. The findings that TEAS modulates mood and anxiety suggest that TEAS could be used as an adjunct in a multimodal therapy program to treat cocaine and cannabis dependence if confirmed in a full randomized controlled clinical trial.
Flowchart of the study. The flow chart of the study was divided into stages: dataset generation, dataset quality control, computation of dual acupoint’s similarity, and statistical analysis.
Similarity maps of channel acupoints calculated based on dermatome information, traditional actions, and contemporary indications. Top row: Similarity maps of dermatomes (left), traditional actions (middle), and contemporary indications (right) arranged in top-down sequence of dermatomes and channels. Bottom row: Similarity maps of dermatomes (left), traditional actions (middle), and contemporary indications (right) arranged after shuffling. Grey-scale colours indicate variations from low (blackish) to high (whitish) values of similarity.
Abbreviations AD: Anno domini; B: Number of replications in resampling; BC: Before Christ; BSA: Body surface area; C 361,361 : Similarity matrix of variable contemporary indications; CI 95%: 95% confidence interval; CSV: Comma separated value; D 361,361 : Similarity matrix of variable dermatomes; J i, j : Jaccard coefficient of similarity between acupoints i and j; N i : Number of terms contained in acupoint i; N ij : Number of terms contained in both acupoints; N j : Number of terms contained in acupoint j; T 361,361 : Similarity matrix of variable traditional actions; γ: Goodman-Kruskal gamma; γ*2: Squared value of the variant of the Goodman-Kruskal gamma.
Analysis of the relationship between the nervous system anatomy and the therapeutic characteristics of all acupuncture points in the channel network may provide new insights on the physiological mechanisms underlying acupuncture stimulation for prevention, treatment, and rehabilitation purposes. This study investigates the association between the similarity of acupoints' dermatomes, traditional actions, and contemporary indications. Channel acupoints had their characteristics annotated from a literature review of four topographic atlases of Chinese medicine and one atlas of human anatomy: initials of the channel's name (n = 14), sequential number in the channel (n = 67), acupoint's name (n = 361), dermatomes related to perpendicular needle insertion (n = 31), traditional actions (n = 848), and contemporary indications (n = 1143). Jaccard's similarity coefficient quantified the similarities between dual acupoints. All dual acupoints were evaluated to generate similarity matrices for each nominal variable. Cross-tables were generated by simultaneous classification of variables into levels of similarity with respect to: dermatomes versus traditional actions, dermatomes versus contemporary indications, and traditional actions versus contemporary indications. Goodman-Kruskal gamma and Rousson gamma*2 were calculated based on cross-tables, bootstrap and permutated samples to evaluate the association and determination coefficient between variables, respectively. Significant associations were observed between levels of similarities of dermatomes and traditional actions (gamma = 0.542; P < 0.001), dermatomes and contemporary indications (gamma = 0.657; P < 0.001), and traditional actions and contemporary indications (gamma = 0.716; P < 0.001). Similarities of dermatomes explained 16% of the variance of traditional actions and 25% of contemporary indications. Traditional actions explained 30% of the variance of contemporary indications. The association between traditional actions and contemporary indications was the highest one (gamma = 0.716, 95% confidence interval (95% CI) = [0.715; 0.719]), followed by the association between dermatomes and contemporary indications (gamma = 0.622, 95% CI = [0.621; 0.623]), and between dermatomes and traditional actions (gamma = 0.446, 95% CI = [0.444; 0.447]), all with P < 0.001. The similarity of dermatomes between dual acupoints partially determined the similarity of traditional actions and contemporary indications, therefore dermatomes partially determine the therapeutic efficacy of acupuncture.
Trial quality of randomized controlled trials with sham acupuncture control
Flow chart showing the retrieval process of clinical trial reports included in the systematic review.
The results of many clinical trials and experimental studies regarding acupoint specificity are contradictory. This review aims to investigate whether a difference in efficacy exists between ordinary acupuncture on specific acupoints and sham acupuncture controls on non-acupoints or on irrelevant acupoints. Databases including Medline, Embase, AMED and Chinese Biomedical Database were searched to identify randomized controlled trials published between 1998 and 2009 that compared traditional body acupuncture on acupoints with sham acupuncture controls on irrelevant acupoints or non-acupoints with the same needling depth. The Cochrane Collaboration's tool for assessing risk of bias was employed to address the quality of the included trials. Twelve acupuncture clinical trials with sham acupuncture controls were identified and included in the review. The conditions treated varied. Half of the included trials had positive results on the primary outcomes and demonstrated acupoint specificity. However, among those six trials (total sample size: 985) with low risk of bias, five trials (sample size: 940) showed no statistically significant difference between proper and sham acupuncture treatments. This review did not demonstrate the existence of acupoint specificity. Further clinical trials with larger sample sizes, optimal acupuncture treatment protocols and appropriate sham acupuncture controls are required to resolve this important issue.
Schematic comparison of the types of needles used in the experiment.
Schematic presentation of the assembly used for the injection at an angle (α) of 45° with the line XY. The point Y was positioned on the Zusanli acupoint, and the assembly was fixed by experimenter fingers against the animal leg. The line ZW, parallel to the line XY, crosses a point 5 mm distal to Y.
Time course of the changes induced by the sham (EA sham) or real electroacupuncture (EA). Anesthesia was provided during the time indicated by a horizontal dashed arrow. EA was applied for 20 min (solid line) using the needle types numbered as in A. Points are means (± SD) of six rats per group. P < 0.05 compared to sham EA (*), sham EA, EA (I) and EA (V) (+), or any other group (#), using the Bonferroni post hoc test.
The changes on the tail-flick latency or paw flinches produced by formalin in the rats. (A) Time course of the changes induced by the injection (arrow) of saline (0.10 μL) into the Zusanli acupoint at 5 mm below the skin surface or 1% formalin (0.10 μL) into sites indicated in the inset on the tail-flick latency. (B) Time course of the changes induced by intraplantar injection of 1% or 2.5% formalin (0.10 μL) at time 0 on the cumulative flinches of the rats. (C) Time course of the changes induced by an intraplantar injection of saline (0.10 μL), 1% or 2.5% formalin (0.10 μL) on the tail-flick latency of the rats. Points are means (± SD) of six rats per group. P < 0.05 compared to saline (*) or any other group (#), using the Bonferroni post hoc test.
The stimulation of acupoints along the meridians, but not the non-acupoints outside of the meridians, produces analgesia. Although the acupoint is defined at the body surface, the exact location of the acupoints is not known. This study aims to examine whether the intensity and duration of the analgesic effect of electroacupuncture (EA) at the Zusanli (ST36) and Sanynjiao acupoints (SP6) change according to the depth of the stimulation. Ninety-six male Wistar rats classified as responders were arbitrarily allocated into 16 groups of six rats each. Six groups received EA with uninsulated acupuncture needles (type I) or needles that were immersed in varnish and had the varnish circularly peeled 0.2 mm from the tip (type II), 0.2 mm at 3 mm (type III) or 5 mm (type IV) from the tip, or 0.2 mm at 5 and 1 mm from the tip (type V), or EA sham for 20 min. Five groups received injection of formalin into the acupoint bilaterally at 5 mm or 1 mm deep into ST36, 5 mm below ST36 but inserting the needle at 45° to the skin surface, or 5 mm deep into non-acupoints. The remaining groups received intraplantar injection of saline, 1% or 2.5% formalin. The analgesic effects were measured by the rat tail-flick test. The bilateral stimulation of ST36 and SP6 by uninsulated or insulated needles produced analgesia in the rat tail-flick test. The stronger and longer lasting effects occurred after EA with the types I and V needles, or injection of formalin 5 mm deep into ST36. The remaining needles produced weaker and shorter lasting effects. Slow analgesic effect also occurred after formalin injection at 1 mm or 5 mm below ST36 by inserting the needle at 45° to the skin surface. The experimental results suggest that the efficacy of the EA stimulation depends on the spatial distribution of the current density under the needling surface rather than only the acupoint or the depth of needling.
Demographic characteristics of the participants
Percent improvements in the VAS, physical function, emotional function and QoL
Mean and SD scores of the VAS, fatigue, and general health status/QoL
Mean scores of the general health status/QoL, VAS, and fatigue.
Many cancer patients seek complementary and alternative medicine (CAM) including acupuncture to manage their cancer-related symptoms or side effects of treatments. Acupuncture is used to manage cancer pain and improve quality of life (QoL). This study aimed to conduct a preliminary study on a case series to evaluate the feasibility of acupuncture for treating cancer pain and to collect preliminary data on the effectiveness of acupuncture in treating cancer pain and improving QoL. A semi-standardized acupuncture treatment comprising one to three treatment sessions (20--30 minutes per session) per week for 8 weeks was provided by four licensed acupuncturists, who had more than 5 years of clinical experience, at the University Health Center. The European Organization for Research and Treatment of Cancer Quality of Life Questionnaire (EORTC QLQ-C3) and a visual analogue scale (VAS) for pain rating were used as the outcome measures to assess pain and QoL. Data were collected at baseline, immediately after 2, 4, 6, and 8 weeks of treatment and at 4 weeks after treatment completion (week 12). Two males and five females with a median age of 66 years (range: 44--71 years) completed the study. For the VAS, the percentage of improvement ranged between 18% and 95%. The baseline mean raw score was reduced from 51 mm to 36 mm at the end of week 8 and to 23 mm at the end of week 12. The percentage of overall QoL improvement ranged between 20% and 100%. The mean raw score for QoL improved with time. The baseline score was increased from 55 to 69 at the end of treatment (week 8) and to 73 after the follow-up (week 12). This pilot study on a case series showed that acupuncture might be beneficial for reducing pain and improving QoL in cancer patients.
Reported respondent rates across recent trials of migraine treated with various interventions. Respondents were defined as those who reported reduction of pain. The figure was modified from a PowerPoint presentation [6] with the permission of Dr M Cummings.
Reported respondent rates across recent trials of low back pain treated with various interventions. Respondents were defined as those who reported increased function. The figure was modified from a PowerPoint presentation [6] with the permission of Dr M Cummings.
Reported respondent rates across recent trials of knee osteoarthritis pain treated with various interventions. Respondents were defined as those who reported increased function. The figure was modified from a PowerPoint presentation [6] with the permission of Dr M Cummings.
Placebo-control of acupuncture is used to evaluate and distinguish between the specific effects and the non-specific ones. During 'true' acupuncture treatment in general, the needles are inserted into acupoints and stimulated until deqi is evoked. In contrast, during placebo acupuncture, the needles are inserted into non-acupoints and/or superficially (so-called minimal acupuncture). A sham acupuncture needle with a blunt tip may be used in placebo acupuncture. Both minimal acupuncture and the placebo acupuncture with the sham acupuncture needle touching the skin would evoke activity in cutaneous afferent nerves. This afferent nerve activity has pronounced effects on the functional connectivity in the brain resulting in a 'limbic touch response'. Clinical studies showed that both acupuncture and minimal acupuncture procedures induced significant alleviation of migraine and that both procedures were equally effective. In other conditions such as low back pain and knee osteoarthritis, acupuncture was found to be more potent than minimal acupuncture and conventional non-acupuncture treatment. It is probable that the responses to 'true' acupuncture and minimal acupuncture are dependent on the aetiology of the pain. Furthermore, patients and healthy individuals may have different responses. In this paper, we argue that minimal acupuncture is not valid as an inert placebo-control despite its conceptual brilliance.
Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) scores
Patient participation flow in the study.
The present study tests whether a combined treatment of acupuncture and transcutaneous electrical nerve stimulation (TENS) is more effective than acupuncture or TENS alone for treating knee osteoarthritis (OA). Thirty-two patients with knee OA were randomly allocated to four groups. The acupuncture group (ACP) received only acupuncture treatment at selected acupoints for knee pain; the TENS group (TENS) received only TENS treatment at pain areas; the acupuncture and TENS group (A&T) received both acupuncture and TENS treatments; the control group (CT) received topical poultice (only when necessary). Each group received specific weekly treatment five times during the study. Outcome measures were pain intensity in a visual analogue scale (VAS) and knee function in terms of the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC). The ACP, TENS and A&T groups reported lower VAS and WOMAC scores than the control group. Significant reduction in pain intensity (P = 0.039) and significant improvement in knee function (P = 0.008) were shown in the A&T group. Combined acupuncture and TENS treatment was effective in pain relief and knee function improvement for the sampled patients suffering from knee OA.
Changes in pressure pain threshold (PPT) values after acupuncture. Pressure pain threshold was measured with a finger type pressure algometer consisting of a strain gage and a processing unit. Changes in the resistance caused by strain in the gage was converted to changes in the voltage via Wheatstone bride in the processing unit which enables quantitative indication of the pressure as an arbitrary units (AU). A significant decrease in the PPT two days after exercise was found in all groups while recovery was observed after application of acupuncture at the skin and the muscle with statistical significance when compared with that in the control at the end of the trial. (P = 0.020 and P < 0.001 for skin and muscle respectively). Asterisks indicate significant differences compared with two days after the exercise (* P < 0.05, ** P < 0.001). Data are shown as mean ± SD.
Changes in electrical pain threshold (EPT) values after acupuncture. Acupuncture to ipsilateral muscle (muscle group) only increased the EPT of fascia significantly (P = 0.028), but skin (skin group) and non-segmental muscle (non-segmental group) had no effect at all tissues (in terms of depths). Asterisk indicates significant differences compared with the control group (* P < 0.05). Data are shown as mean ± SD.
While evidence supports efficacy of acupuncture and/or dry needling in treating musculoskeletal pain, it is unclear which needling method is most effective. This study aims to determine the effects of depth of needle penetration on muscle pain. A total of 22 healthy volunteers performed repeated eccentric contractions to induce muscle soreness in their extensor digital muscle. Subjects were assigned randomly to four groups, namely control group, skin group (depth of 3 mm: the extensor digital muscle), muscle group (depth of 10 mm: the extensor digital muscle) and non-segmental group (depth of 10 mm: the anterior tibial muscle). Pressure pain threshold and electrical pain threshold of the skin, fascia and muscle were measured at a point 20 mm distal to the maximum tender point on the second day after the exercise. Pressure pain thresholds of skin group (depth of 3 mm: the extensor digital muscle) and muscle group (depth of 10 mm: the extensor digital muscle) were significantly higher than the control group, whereas the electrical pain threshold at fascia of muscle group (depth of 10 mm: the extensor digital muscle) was a significantly higher than control group; however, there was no significant difference between the control and other groups. The present study shows that acupuncture stimulation of muscle increases the PPT and EPT of fascia. The depth of needle penetration is important for the relief of muscle pain.
Acupuncture is often used to treat fibromyalgia (FM), but it remains unclear whether acupuncture is effective. This study aims to evaluate the effects of acupuncture on pain and quality of life (QoL) in FM patients. Sixteen patients (13 women and 3 men aged 25-63 years) suffering from FM were randomised into two groups: group A (n = 8) received five acupuncture treatments after the fifth week and group B received ten acupuncture treatments. Outcome measures used in this study were pain intensity (visual analogue scale, VAS) and the fibromyalgia impact questionnaire (FIQ). After the fifth week, pain intensity (U = 25.0; P = 0.022) in group B decreased and QoL (U = 24.5; P = 0.026) improved compared to group A. The present study suggests that acupuncture treatment is effective to relieve pain for FM patients in terms of QoL and FIQ.
Characteristics of included trials of moxibustion, acupuncture and other acupoint stimulation for breech presentation
Efficacy of moxibustion, acupuncture or other acupoint stimulations for the correction of breech presentation
Process of trial identification and selection.
Moxibustion, acupuncture and other acupoint stimulations are commonly used for the correction of breech presentation. This systematic review aims to evaluate the efficacy and safety of moxibustion and other acupoint stimulations to treat breech presentation. We included randomized controlled trials (RCTs) and controlled clinical trials (CCTs) on moxibustion, acupuncture or any other acupoint stimulating methods for breech presentation in pregnant women. All searches in PubMed, the Cochrane Library (2008 Issue 2), China National Knowledge Information (CNKI), Chinese Scientific Journal Database (VIP) and WanFang Database ended in July 2008. Two authors extracted and analyzed the data independently. Ten RCTs involving 2090 participants and seven CCTs involving 1409 participants were included in the present study. Meta-analysis showed significant differences between moxibustion and no treatment (RR 1.35, 95% CI 1.20 to 1.51; 3 RCTs). Comparison between moxibustion and knee-chest position did not show significant differences (RR 1.30, 95% CI 0.95 to 1.79; 3 RCTs). Moxibustion plus other therapeutic methods showed significant beneficial effects (RR 1.36, 95% CI 1.21 to 1.54; 2 RCTs). Laser stimulation was more effective than assuming the knee-chest position plus pelvis rotating. Moxibustion was more effective than no treatment (RR 1.29, 95% CI 1.17 to 1.42; 2 CCTs) but was not more effective than the knee-chest position treatment (RR 1.22, 95% CI 1.11 to 1.34; 2 CCTs). Laser stimulation at Zhiyin (BL67) was more effective than the knee-chest position treatment (RR 1.30, 95% CI 1.10 to 1.54; 2 CCTs,). Moxibustion, acupuncture and laser acupoint stimulation tend to be effective in the correction of breech presentation.
Pain intensity scores in visual analog scale (VAS)
Subject participation flow in the study.
Acupuncture is used to reduce inflammation and decrease pain in delayed onset muscle soreness (DOMS). This study investigates the efficacy of acupuncture on the symptoms of DOMS. Thirty subjects were assigned randomly to there groups, namely the control, non-tender point and tender point groups. Measurement of pain with full elbow flexion was used as indices of efficacy. Measurements were taken before and after exercise, immediately after treatment and seven days after treatment. Significant differences in visual analog scores for pain were found between the control group and tender point group immediately after treatment and three days after exercise (P < 0.05, Dunnetts multiple test). The results show that tender point acupuncture relieves muscle pain of DOMS.
Baseline participant characteristics
Participants flow in the study.
VAS scores for pain intensity. (A) Comparison between control and AC, BA or BA+AC. (B) Comparison between AC, BA and BA+AC. * P < 0.05. ** P < 0.01. *** P < 0.001.
RDQ scores for pain disability. (A) Comparison between control and AC, BA or BA+AC. (B) Comparison between AC, BA and BA+AC. * P < 0.05. *** P < 0.001.
Chronic non-specific low back pain (LBP) is a prevalent (80%) and multi-dimensional illness. This study aims to test whether acupuncture, baclofen, or combined treatment with acupuncture and baclofen alleviates symptoms of non-specific chronic LBP in men. Eight-four (84) men aged 50-60 years with non-specific chronic LBP were randomly assigned to four groups: the baclofen group received only baclofen (30 mg/day); the acupuncture group received only acupuncture at selected acupoints; the acupuncture + baclofen group received combined treatment with acupuncture and baclofen treatments; and the control group received no pain reduction treatment. After five weeks of treatment, visual analogue scale (VAS) and self-reported pain disability with the Roland-Morris Disability Questionnaire (RDQ) were conducted for outcome measures. After treatment, the baclofen, acupuncture and acupuncture + baclofen groups all had lower VAS and RDQ scores. Significantly higher reduction and improvement in VAS and RDQ scores were found in the acupuncture and acupuncture + baclofen groups compared to the baclofen group. The present study indicates that the combined treatment of acupuncture and baclofen is more effective than baclofen treatment alone to reduce pain in patients with non-specific chronic LBP. ACTRN12609000698279.
Top-cited authors
Qing-Wen Zhang
  • University of Macau
Shilin Chen
  • China Academy of Chinese Medical Sciences
Priya Thambi Thekekara
  • Baselius College Kottayam
Sabu Mc
  • Mookambika College of Pharmaceutical Sciences and Research, Muvattupuzha, Kerala, India
Zhangfeng Zhong
  • University of Macau