In this paper, electronic nose (E-nose) was used to discriminate the 11 Chinese Materia Medica (CMM) from Umbelliferae by the difference of their odors. The E-nose generated data were analyzed by discriminant function analysis (DFA) and the responses of 18 sensors of E-nose were evaluated by CA analysis. Results showed that a rapid evaluation of complex response of the samples could be obtained, in combination with DFA, SQC and the CA analysis of the E-nose was given better results in the recognition values of the odor of the 11 CMM. All the 11 CMM could be distinguished by E-nose coupled with DFA, sensor 2, 3, 4, 5, 11, 13 and 15 were found to be able to better discriminate between the CMM samples. The CMM from Umbelliferae produced from different areas and processed with different methods could be distinguished by the E-nose, too. The results of the similarity of fingerprints of the E-nose are fitted with the TCM records about the property (yaoxing), channel tropism (guijing), function and usage of the CMM. The E-nose is a technology that can reflect the holistic odor of a CMM and is relevant to the TCM doctor’s practical identification. The odor of CMM can be expressed by objective data instead of subjective sense by human nose. Based on the sensor`s intensity of E-nose, the fingerprint of a CMM can be established, too. Although the E-nose has so many advantages, only use E-nose technology is not enough to control the quality of a CMM. It must be combined with the other macroscopic discriminating technology, such as the E-tongue, the E-eye, to have a holistic evaluation of a CMM.
A subchronic oral toxicity was conducted to evaluate the safety of total flavones of E. leptorrhizum Stearn in Sprague-Dawley rats. The test article was administered once daily by gavage in male and female rats at dose levels of 24, 48, and 96 mg/kg body weight/day for 180 days. 90 and 180 days after administration, ten and tweedy animals (each half of male and female) of each group were tested. 28 days after withdrawal, five male and female rats were tested. There were no significant toxicological changes shown in daily clinical signs, body weight, food consumption, hematology parameters, blood biochemistry, organ weights and histopathological examination except leukocyte differential count. It was concluded that the no-observed-effect level (NOEL) for total flavones of E. leptorrhizum Stearn was >96 mg/kg in SD rats.
Aims: To study RBL-2H3 cell degranulation phenomena induced by some TCMIs through cell morphological and ultra-structural observation, released enzyme activity and establish RBL-2H3 cell degranulation test indicated by β- hexosaminidase activity as a method to evaluate TCMIs at nonclinical stage. Methods: RBL-2H3 cells were used to study the degranulation by co-culture with positive control C48/80 and some TCMIs through morphological and ultra-structure observation, β-hexosaminidase activity detection. RBL-2H3 cell degranulation test was established to detect β-hexosaminidase activity caused by 17 kinds of TCMIs and their ingredients. The cytotoxicity effect of some TCMIs on both RBL 2H3 and BRL cells was measured by CCK-8 assay. Results: Toluidine blue staining and ultra-structure of electronic microscope observation of treated RBL-2H3 cells showed degranulation morphologically. Detection of β-hexosaminidase activity in the supernatant of treated cells showed some TCMIs had elevated enzyme release rates. Further analysis of the ingredients and compound in Tanreqing Injection and Shengmai Injection showed Scutellaria baicalensis Georgi in Tanreqing Injection, Red ginseng and Fructus Schisandrae Chinensis in Shengmai Injection were responsible to the degranulation of RBL-2H3 cells. Osmotic pressures and pH influenced RBL-2H3 degranulation. High Osmotic pressure of Tanreqing Injection and low pH of chlorogenic acid at 2.5 and 5.0 mmol/L congcentration might be responsible to high β-hexosaminidase activity. Most of the TCMIs inducing degranulation had cytotoxicity effect for both RBL-2H3 and BRL cells, but some TCMIs inducing degranulation had no cytotoxicity effect. Conclusion: Some TCMIs can induce degranulation of RBL-2H3 cells; RBL-2H3 cell degranulation test can be used in non-clinical stage to detect the risk causing anaphylactoid reactions. Osmotic pressures and pH influenced RBL-2H3 degranulation, and they should be measured before testing. The mechanism of degranulation caused by some TCMIs is cytotoxic, and some are non-cytotoxic and may be through exicytosis.
Superoxide dismutase (SOD) is an important antioxidant enzyme present in all oxygen-metabolizing cells. The effect of different wavelengths on superoxide dismutase (SOD) has been reported previously which has shown to have a remarkable effect on immune system. Thus, after presenting solid scientific reasoning behind this therapeutic mode of treatment pertaining to the activity of SOD in the presence of 644 nm radiation, a randomised controlled clinical trial has been carried out which builds a unique relationship of 644 nm irradiated SOD with the elimination of free radicals and hence the enhancement of immunity, not only for curative treatment but also as a preventive tool against all those diseases related to the elimination of free radicals to get better immune system of the human body. This study was conducted in the department of Gynecology and obstetrics at Al-Khidmat Teaching Hospital Mansoorah, Lahore, Pakistan.
The in vitro antiprotozoal and cytotoxic activity of the aqueous extract, the 80% methanol extract, and its different soluble fractions and subfractions from Brucea sumatrana seeds were assessed against two Trypanosoma (T. cruzi and T. brucei brucei), Leishmania infantum and chloroquine and pyrimethanine-resistant K1strain of P. falciparum and against MRC-5 cell-lines respectively. Results indicated that the 80% methanol extract showed a cytotoxic effect against MRC-5 cell lines with CC50 value of 0.54 μg/ml. It however exhibited pronounced and non selective activity against T. cruzi (IC50 = 1.52 μg/ml, SI = 0.03) and L. infantum (IC50 = 2.41 μg/ml, SI = 0.22). It however displayed pronounced and selective effect against T. brucei brucei (IC50 2.16) and chloroquine and pyrimethamine-resistant K1 strain of P. falciparum (IC50 2.16). All soluble fractions and subfractions from the partition of the 80% methanol extract were found to exhibit an antiprotozoal activity with IC50 values ranging from T .cruzi, T. b. brucei, L. infantum and chloroquine and pyrimethamine-resistant K1 strain of P. falciparum with IC50 values of 0.33, 81, 81 and >81 respectively. The chloroform soluble fraction rich in alkaloid was cytotoxic against MRC-5 cell lines (CC50 = 27.09 μg/ml) and showed good activity against T. b. brucei (IC50 = 8.36 and SI = 3.24) and moderate activity against T. cruzi, L. infantum and chloroquine-pyrimethane-resistant K1 strain of P. falciparum (20 50 50 = 1.55 and 0.43 μg/ml respectively), they however displayed pronounced antiprotozoal activity against T. cruzi, T. b. brucei and chloroquine and pyrimethamine-resistant K1 strain of P. falciparum with IC50 values ranging from P. falciparum (SI = >6.2 and >1.72 respectively). These extracts however showed good and low activity respectively against L. infantum (IC50 = 24.05 and 6.82 μg/ml respectively).
Results from the in
vitro evaluation of the antiparasitaire activity of the aqueous extract,
the 80% methanol extract and its fractions from the leaves of Brucea sumatrana against Trypanosoma brucei brucei, T. cruzi, Leishmania infantum, the multidrug-resistant K1 and
chloroquine-sensitive NF54 strains of Plasmodium
falciparum indicated that all samples from the leaves extract presented
interesting antiparasitaire activity at different extents. The 80% methanol
extract, its chloroform acid, petroleum ether and 80% methanol soluble
fractions and the aqueous extract exhibited strong activity against Trypanosoma b. brucei, T. cruzi, L. infantum and the multidrug-resistant K1 strain of P. falciparum with IC50 values from P. falciparum with IC50 values ranging from 50)
50 was estimated to be greater than 5 g/kg. In addition, it
did not significantly modify the concentration levels of some evaluated
biochemical and hematological
parameters in treated rats. These results constitute a scientific validation
supporting and justifying the traditional use of the leaves of B. sumatrana for the treatment of
malaria, sleeping sickness and at some extent Chagas disease.
Tilia species have been used in Asia, Europe and in America to treat anxiety and also for the treatment of colds and in-flammation. The oxygen reactive species (ROS) (hydrogen peroxide (H 2 O 2) and the superoxide anion (O 2) are involve in the balance cell proliferation/death in lymphocytes. It was reported the presence of flavonoids in Tilia species which possess antioxidant properties. The aim of this study was to determine comparatively the effect of an aqueous (AE) and ethanol (E) extract from Tilia x viridis, on the proliferation of tumoral and normal concanavalin A stimulated murine lymphocytes in relation to antioxidant activities such as peroxidase (Px), catalase (CAT) and superoxide dismutase (SOD) activities involves in H 2 O 2 modulation. Also a phytochemical pattern of the two extracts in relation to flavonoids content was determined. Both extracts presented antiproliferative action on both type of lymphocytes but E was more selective on the tumoral lymphocytes inhibition (EC 50 (µg/ml, Mean ±SEM) (tumoral): 50 ± 4; EC 50 ; (normal lympho-cytes): 323 ± 20); this action was related to a high polyphenols content (150 ± 10 mg/g extract) and high "per se" SOD and low Px activities. In conclusion, the extracts could be a source of antioxidant compounds which contribute to a se-lective antiproliferative action on tumoral cells, acting through the modulation of H 2 O 2 levels.
Tetrandrine (1 μM), a bis-benzylisoquinoline alkaloid isolated from Stephania tetrandra S Moore, significantly decreased tumor necrosis factor alpha (TNFα; 10 ng/ml)-induced increase in the number of micro vessels that budded from cultured rat choroidal explants. Tetrandrine also decreased the TNFα-induced increase in the number of cells composing the microvessels. Ammonium pyrrolidine dithiocarbamate (APDC; 0.1-0.3 μM), an inhibitor of nuclear factor-κB (NF-κB), decreased the TNFα-induced increase in the number of microvessels in a concentration-dependent manner. TNFα increased the phosphorylation and degradation of inhibitor of NF-κB (IκBα), as well as increasing the DNA-binding activity of NF-κB in choroidal explants. TNFα induced an increase of vascular endothelial growth factor (VEGF)-A mRNA, but not VEGF-C mRNA or VEGF-D mRNA. TNFα-induced angiogenic action was inhibited by treatment of VEGF-A antibody in cultured choroidal capillaries. Tetrandrine inhibited the TNFα-induced increases of phosphorylation and degradation of IκBα, and reduced the TNFα-induced increase of DNA-binding activity of NF-κB in choroidal explants. In conclusion, tetrandrine inhibits TNFα-induced activation of NF-κB in the choroidal capillaries via inhibition of TNFα-induced phosphorylation of IκBα.
The structure and size range of an acupoint were investigated thoroughly according to the expositions of ancient classics. It was revealed that the ambiguous way of thinking by the ancient Chinese had a great impact on the formation and development of the acupoint-concept. Hence all descriptions of an acupoint’s structure and size range have the characteristics of ambiguity. The structure and size range of an acupoint are determined not only by the outlook of depression and blood vessel areas, but also its relationship with other acupoints in its vicinity. The different manipulations of puncturing recorded in the Yellow Emperor’s Canon of Medicine also show the ambiguous nature of the structure and size range of an acupoint. In theory, an acupoint has been characterized as various forms, which should not be limited to a mere round dot shape.
Objective: The objective is to observe the treatment effect of electro-acupuncture (EA) on core circadian clock gene Per2 and Bmal1 expression in hypothalamus of sleep-deprivation (SD) rats. Methods: Thirty-two Wistar male rats were randomly divided into 4 groups. Mice in the blank control group did not receive any treatment; the remaining groups were applied with para-chloro-phenylalanine (PCPA) 300 mg/kg intraperitoneal injection for 2 days. Diazepam group received intraperitoneal injection of Diazepam (0.9 mg/kg, i.p.) one time a day for 5 days, while M group was treated with saline (0.9 mg/kg, i.p.) at the same time. Rats in EA group were given EA treatment , 20 minutes, once a day for 5 days, and rats in remaining groups were put into fixation-machine for the same time everyday, lasting for 5 days. Rats were sacrificed after anesthesia at the 8th day. Real-time PCR was adopted to detect the expression in clock gene Per2 and Bmal1 of each group. Results: Compared with blank control group, the expression of Per2 was significant decreased in PCPA model group (P < 0.05), the expression of Bmal1 was increased in PCPA model group (P < 0.05). Compared with PCPA model group, the expression of Per2 were significant enhanced in EA group and Diazepam group (P < 0.05). Simultaneously, compared with PCPA group, the expression of Bmal1 was no statistically significant in EA group and Diazepam groups (P > 0.05). Conclusion: EA can significant up-regulate the expression of Per2 in SD rats, and down-regulate gene Bmal1 expression, and benefiting the weight of rats. Thus, EA is a potentially promising intervention to treat sleep-deprivation.
Skeletal muscle fatigue is a common symptom in various diseases, works and exercises. These were generally induced by neuron, metabolic conditions, overused muscle, and stress. But, there have been few principles about it. Many re-searchers have reported that acupuncture therapy has been useful to skeletal muscle fatigue on various diseases and conditions. However, it has never been shown why acupuncture therapy has the effect on skeletal muscle fatigue. The deficiency of carnitine induces fatigue, weakness, and disorder of skeletal muscle. It has showed that acupuncture in-duces the increase of carnitine in skeletal muscle. These findings demonstrated that acupuncture on skeletal muscle fa-tigue could increase carnitine as a possible affection mechanism.
Based on related elaboration of the Yellow Emperor’s Canon of Medicine, this article analyzed and summarized the clinical meaning, application principles and the basic operating methods of Traditional Acupuncture (TA), and demonstrated that the TA is completely different to modern needle stimulation. TA has a specific application background, direct-viewing thinking mode and clear operational connotation. The key of operation in TA is how to grasp and control Qi, which typically reflect the unique image of the Chinese civilization with intuitive perceptual characteristics of thinking. In contrast, modern needle-stimulation uses needles as a stimulus, to activate a series of physical and functional reactions in a body. There have great differences between the two. It was indicated that correctly understanding with the basic principle and specific meaning of TA is very important in acupuncture clinical and research works.
Objective: To investigate effects of Cornus officinalis Total Glycosides (COTG) and Cornus Polysac- charide (CP) on myocardial protection and on expression of mitochondria biogenesis related gene of acute myocardial infarction (AMI) rats, Materials and Methods: Ninety-six SD rats of SPF level were randomly divided into 5 groups: sham operation group, model group, preventive treatment group, COTG treatment group, CP treatment group, and there were 12 cases in each one. By legat- ing the left anterior descending branch of coronary artery method, acute myocardial infarction model was established. The rat of sham operation group and model group was intragastric admi- nistered with physiological saline; other groups were given with corresponding drugs. The cardiac function, the myocardial infarct area, the expression of mitochondrial biogenesis genes such as PGC-1α, PGC-1β, NRF-1mRNA and GSK-3β mRNA, GSK-3β Protein Expression were analyzed. Re- sults: The results revealed that compared with model group, myocardial infarction size, LVDs, LVDd, LVESV, LVEDP, and −dp/dt decreased; LVSP increased in preventive treatment group, COTG treatment group, and CP treatment group (p < 0.05); LVEDV increased in preventive treatment group (p < 0.05), PGC 1 alpha, and PGC 1 beta; the NRF-1 mRNA expression increased in preventive treatment group, COTG treatment group, and CP treatment group (p < 0.05). Compared with CP and COTG treatment group, PGClpha, beta PGC 1, the NRF-1 mRNA expression increased in pre- ventive treatment group (p < 0.05). Compared with the sham operation group, GSK-3 beta mRNA and protein expression increased in model group, preventive treatment group, COTG treatment group, and CP treatment group (p < 0.05). Compared with model group, GSK-3 beta mRNA expres- sion reduced in preventive treatment group, COTG treatment group, and CP treatment group (p < 0.05). Conclusions: Cornus officinalis total glycosides and Cornus polysaccharides can effectively * Corresponding author.
This study aimed to investigate the effects of a compound polysaccharide-based health product (Infinitus Polysac Plus, IPP) on innate and adaptive immunity in mice. Both ex vivo/in vivo mouse models and an in vitro system using cultured mouse splenocytes were adopted for the assessment of innate and adaptive immunity. For the innate immune response, long-term IPP treatment (0.26 and 0.78 g/kg * 20 doses) enhanced the carbon clearance activity and phagocytic rate of macrophages, as well as natural killer cell activity in mice. The IPP-induced increase in natural killer cell activity was accompanied by the suppression of tumor growth in Sarcoma-180 cell-inoculated mice. For the adaptive immune response, while long-term IPP treatment increased the splenocyte index in mice, IPP incubation with mouse splenocytes in vitro potentiated their concanavalin A-stimulated proliferation. Long-term IPP treatment significantly restored the hemolytic activity of serum on sheep red blood cells and dinitrofluorobenezene-induced delayed-type hypersensitivity in sensitized and immunosuppressed mice. In conclusion, the results indicate that long-term IPP treatment produces stimulatory effects on both innate and adaptive immunity in mice.
The 18 crude drugs in Bofutsushosan (BOF: Pulvis ledebouriellae compositae: 防風通聖散) are separated into 6 groups such as diaphoretic, cathartic, antidote, antipyretic, neutralizer and diuretic groups. The effects of single administered BOF and composed crude drugs in 6 groups were investigated on the levels of diabetic parameters (serum glucose, insulin, triglyceride and cholesterol) in streptozotocin-induced diabetic mice. The anti-hyperglycemic action of BOF was depended on Ephedrae Herba, Saposhnikoviae Radix and Schizonepetae Spica in diaphoretic group, Forsythiae Fructus, Saposhnikoviae Radix, Schizonepetae Spica and Cnidii Rhizoma in antidote group, Scutellariae Radix, Gardeniae Fructus and Gypsum Fibrosum in antipyretic group and Paeoniae Radix in neutralizer group. In these crude drugs, Ephedrae Herba, Saposhnikoviae Radix, Schizonepetae Spica, Forsythiae Fructus, Scutellariae Radix, Gypsum Fibrosum and Paeoniae Radix increased serum insulin level, but Cnidii Rhizoma and Gardeniae Fructus did not affect serum insulin level. From these results, it suggested that anti-hyperglycemic action of BOF was through insulin-dependent and insulin independent manners. The lowering effect of BOF on serum triglyceride level was dependent on actions of Platycodi Radix in antidote and diuretic groups and Gardeniae Fructus in antipyretic group. The lowering effect of Gardeniae Fructus was parallel with its anti-hyperglycemic action. The lowering effect of BOF on high serum triglyceride level also included both direct action and indirect action. The reducing effect of BOF on serum cholesterol level was observed together with the actions of Ephedrae Herba and Zingiberis Rhizoma in diaphoretic group, Schizonepetae Spica in diaphoretic and antidote groups and Paeoniae Radix in neutralizer group. The lowering effects of Ephedrae Herba, Schizonepetae Spica and Paeoniae Radix on serum cholesterol level were parallel with their anti-hyperglycemic actions. Zingiberis Rhizoma in diaphoretic group might be direct reducing effect on serum cholesterol level but no serum glucose level. The Ephedrae Herba in diaphoretic group, Schizonepetae Spica in diaphoretic and antidote groups and Paeoniae Radix in neutralizer group might have reduced serum cholesterol level by reducing blood glucose level. From these results, composed crude drugs in 6 groups show various mechanisms in the action of BOF.
The mechanisms of Gardeniae Fructus (GF) for anti-hyperglycemic action were demonstrated in streptozotocin (STZ)-diabetic mice. Six hours after single intraperitoneal administration of GF (300 mg/kg) or H2O into 3 hour-fasted STZ-diabetic mice, glucose and insulin tolerances were assessed by intraperitoneal glucose (1.5 g/kg) tolerance test (IPGTT) and intraperitoneal insulin (0.65 U/kg) tolerance test (IPITT), respectively. Effects of GF on insulin signaling pathways in soleus muscle such as glucose uptake, expression of glucose transporter 4 (GLUT4) in the plasma membrane and phosphorylation of Akt (P-Akt) in cytosolic fraction were examined in STZ-diabetic mice. In IPGTT test, GF significantly accelerated clearance of exogenous glucose and its glucose-lowering action was greater than H2O-treated controlin STZ-diabetic mice. GF also promoted an exogenous glucose-increased insulin level in STZ-diabetic mice. In IPITT test, GF decreased glucose level to the greater extent than H2O-treated control in STZ-diabetic mice. Furthermore, GF significantly decreased high HOMA-IR in STZ-diabetic mice from 21.6 ± 2.4 to 12.4 ± 1.9 (mg/dl × μU/ml). These results implied that GF improved insulin resistance in STZ-diabetic mice. GF increased glucose uptake of soleus muscle 1.5 times greater than H2O-treated control in STZ-diabetic mice. GF enlarged insulin (10 nmol/ml)-increased glucose uptake to 1.8 time-greater. Correspondingly, GF increased expression of GLUT4 in the plasma membrane of soleus muscle to 1.4 time-greater, and P-Akt in the cytosolic fraction of soleus muscle to 1.9 time-greater than those in H2O-treated control. In conclusion, the improvement of GF on insulin resistance is associated with the repair of insulin signaling via P-Akt, GLUT4 and glucose uptake pathway in soleus muscle of STZ-diabetic mice.
Irritable bowel syndrome (IBS) is characterized by chronic visceral hypersensitivity that companied by altered bowel movement. However, the treatment options are very limited. The aim of this study was to investigate effects of electroacupuncture (EA) on visceral hypersensitivity in a rat model of IBS and to explore the underlying mechanisms of EA effects. Visceral hypersensitivity was established by neonatal maternal deprivation (NMD) in male rats on postnatal days 2 - 15. Behavioral experiments were conducted at the age of 7 weeks. Treatment with EA at Zusanli (stomach-36, ST-36) significantly reduced abdominal withdrawal reflex (AWR) scores in NMD rats but not in age-matched healthy control rats. In addition, EA treatment hyperpolarized resting membrane potentials, increased the rheobase and reduced the numbers of action potentials evoked by 2 and 3 times rheobase current stimulation of dorsal root ganglion (DRG) neurons innervating the colon. NMD markedly enhanced expression of TRPV1 in colon related DRGs while EA treatment drastically suppressed the expression of TRPV1 in DRGs of NMD rats. These data suggest that EA treatment produced an analgesic effect, which might be mediated at least in a part by suppression of TRPV1 expression and by inhibition of neuronal excitability of primary sensory neurons in rats with chronic visceral pain.
Rhizoma Menispermi, derived from the rhizoma of Menispermum dauricum DC., is one of the most popular Chinese medicines. However Rhizoma Menispermi is often illegally mixed with other species in the herbal market, including Aristolochia mollissimae Hance, which is toxic to the kidneys and potentially carcinogenic. The use of DNA barcoding to authenticate herbs has improved the management and safety of traditional medicines. In this paper, 49 samples belonging to five species, including 34 samples of M. dauricum, from different locations and herb markets in China were collected and identified using DNA barcoding. The sequences of all 34 samples of Rhizoma Menispermi are highly consistent, with only one site variation in internal transcribed spacer 2 (ITS2) of nuclear ribosomal DNA and no variations in the psbA-trnH region. The intra-specific genetic distance is much smaller than inter-specific one. Phylogenetic analysis shows that both sequences allow the successful identification of all species. Nearest distance and BLAST1 methods for the ITS2 and psbA-trnH regions indicate 100% identification efficiency. Our research shows that DNA barcoding can effectively distinguish Rhizoma Menispermi from its adulterants from both commercial and original samples, which provides a new and reliable way to monitor commercial herbs and to manage the modern medicine market.