[Show abstract][Hide abstract] ABSTRACT: DNA damage repair genes JWA, XRCC1 and BRCA1 were associated with clinical outcomes and could convert the response to the cisplatin-based therapy in some carcinomas. The synergistic effects of JWA, XRCC1 and BRCA1 mRNA expression on personalized therapy remain unknown in advanced esophageal squamous cell carcinoma (ESCC).
We employed quantitative real-time polymerase chain reaction (qPCR) to determine the expression of JWA, XRCC1 and BRCA1 mRNA in paraffin-embedded specimen from 172 patients with advanced ESCC who underwent the first-line cisplatin-or docetaxel-based treatments.
High JWA or XRCC1mRNA expression was correlated with longer median overall survival (mOS) in all the patients (both P < 0.001) or in subgroups with different regimens (all P < 0.05), but not correlated with response rate (RR, all P > 0.05). Multivariate analysis revealed that high JWA (HR 0.22; 95% CI 0.13-0.37; P < 0.001) or XRCC1 (HR 0.36; 95% CI 0.21-0.63; P < 0.001) mRNA expression emerged as the independent prognostic factors for ESCC patients in this cohort. But no significant difference in prognostic efficacy was found between JWA plus XRCC1 and JWA alone through ROC analysis. Further subgroup analysis showed cisplatin-based treatments could improve mOS of patients with low JWA expression (P < 0.05), especially in those with low BRCA1 expression simultaneously (P < 0.001); while in patients with high JWA expression, high BRCA1 mRNA expression was correlated with increased mOS in docetaxel-based treatments (P = 0.044).
JWA, XRCC1and BRCA1 mRNA expression could be used as predictive markers in molecular staging for personalized therapy in patients with advanced ESCC who received first-line cisplatin- or docetaxel-based treatments.
BMC Cancer 04/2015; 15(1):331. DOI:10.1186/s12885-015-1364-0 · 3.32 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Diabetes mellitus is a serious chronic metabolic disorder. To develop novel anti-diabetic drugs from nature sources has always been the focus of research. Red deer (Cervus elaphu Linnaeus) antler is one of the most famous Chinese traditional medicines. We found that the peptides of 5-10 kDa from red deer antlers (PRDA) promoted the growth of cultured rat islet cells. The purpose of this study was to investigate the anti-diabetic actions of PRDA in vivo and purify a pure active peptide. We therefore investigated the hypoglycemic, hypolipidemic and antioxidant effects of PRDA in streptozotocin-induced diabetic mice and isolated a pure anti-diabetic peptide. PRDA, given intraperitoneally (75, 150, or 300 μg/kg), significantly decreased the blood glucose levels, significantly increased the insulin concentrations, and remarkably improved the lipid metabolism in the diabetic mice. PRDA significantly increased the superoxide dismutase activity, catalase activity and the total antioxidant capacity in the serum and liver, and simultaneously decreased the malondialdehyde levels. The activities of hexokinase and pyruvate kinase, two important enzymes involved in glucose utilization, were also significantly increased in the liver of the PRDA-treated diabetic mice. Moreover, a novel anti-diabetic peptide isolated from PRDA significantly promoted the viability of cultured rat insulinoma cells. The molecular mass of the purified peptide was 7064.8 Da under mass spectrometry, and its N-terminal amino acid sequence was identified as LSPFTTKTYFPHFDLSHGSA. Thus, PRDA may be useful in managing the hyperglycemia, hyperlipidemia, and oxidative stress in diabetes, and the anti-diabetic peptide is a promising drug for the treatment of diabetes.
The Tohoku Journal of Experimental Medicine 01/2015; 236(1):71-9. DOI:10.1620/tjem.236.71 · 1.28 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Diabetes mellitus is a group of complicated metabolic disorders characterized by high blood glucose level and inappropriate insulin secreting capacity due to decreased glucose metabolism and pancreatic β cell mass or dysfunction of β cells. Thus, improving glucose metabolism and preserving β cell mass and function might be useful for the treatment of diabetes. In this study, a novel acidic polysaccharide LBP-s-1 extracted from Lycium barbarum L. was obtained by purification using macroporous resin and ion-exchanged column. Monosaccharide composition analysis indicated that LBP-s-1 was comprised of rhamnose, arabinose, xylose, mannose, glucose, galactose, galacturonic acid in the molar ratio of 1.00:8.34:1.25:1.26:1.91:7.05:15.28. The preliminary structure features of LBP-s-1 were investigated by FT-IR, (1)H NMR and (13)C NMR. In vitro and in vivo hypoglycemic experiments showed that LBP-s-1 had significant hypoglycemic effects and insulin-sensitizing activity through increasing glucose metabolism and insulin secretion and promoting pancreatic β cell proliferation. Preliminary mechanisms were also elucidated.
[Show abstract][Hide abstract] ABSTRACT: Bacillus subtilis was engineered into an efficient hyaluronic acid (HA) producer by introducing two inducible artificial operons carrying HA synthase gene from Pasteurella multocida and precursor genes encoding enzymes involved in synthesis of the sugar precursors. A two-stage induction strategy was established for metabolic engineering of recombinant B. subtilis to efficiently produce uniform HA with controlled molecular weights. Strain TPG223 produced larger HA molecules (yield=6.8g/L; molecular weight=4.5MDa) than strain PG6181 (yield=2.4g/L; molecular weight=13KDa), indicating that the enzymes involved in the synthesis of UDP-glucuronic acid are essential for HA biosynthesis. Strain TPG223 was able to synthesize HA molecules ranging in molecular weight from 8KDa to 5.4MDa indicating that size control is achievable in vivo through appropriate tools. The work reported here not only advanced mechanisms research of size control in vivo, but also could be an attractive alternative for commercial preparation of uniform size-defined HA.
[Show abstract][Hide abstract] ABSTRACT: The molecular backgrounds that determine therapeutic effectiveness in esophageal cancer remain largely unknown. Breast cancer susceptibility gene 1 (BRCA1) expression has been found to switch the response to cisplatin- or paclitaxel-based chemotherapy. It remains unclear how variations in BRCA1 expression influence clinical outcomes in esophageal cancer.
Quantitative real-time polymerase chain reaction (qPCR) was performed to examine BRCA1 mRNA expressions in paraffin-embedded specimens from 144 patients with advanced or metastatic esophageal squamous cell carcinoma who received cisplatin- or docetaxel-based first-line treatments.
Low BRCA1 mRNA expression correlated with increased response rate (RR; P = 0.025 and 0.017, respectively) and median overall survival (mOS; P = 0.002 and P<0.001, respectively) in cisplatin-based chemotherapy or chemoradiotherapy group and also correlated with decreased RR (P = 0.017 and 0.024, respectively) and mOS (both P<0.001) in docetaxel-based chemotherapy or chemoradiotherapy group. Multivariate analysis revealed that low BRCA1 expression was an independent prognostic factor in cisplatin-based chemotherapy (HR 0.29; 95%CI 0.12-0.71; P = 0.007) or chemoradiotherapy (HR 0.12; 95%CI 0.04-0.37; P<0.001) group and higher risk for mortality in docetaxel-based chemotherapy (HR 5.02; 95%CI 2.05-12.28; P<0.001) or chemoradiotherapy (HR 7.02; 95%CI 2.37-27.77; P<0.001) group.
BRCA1 mRNA expression could be used as a predictive and prognostic marker in esophageal cancer who underwent first-line cisplatin- or docetaxel-based treatments.
PLoS ONE 01/2013; 8(1):e52589. DOI:10.1371/journal.pone.0052589 · 3.23 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: To compare the pulmonary absorption characteristics of two insulin solutions-humalog (insulin lispro) and Novolin R (Biosynthetic Human insulin) with in vivo and in vitro methods.
Investigate the pharmacodynamics in Sprague Dawley (SD) rat model (in vivo studies) and permeability across Rana catesbeiana pulmonary membrane (in vitro studies) of Biosynthetic Human insulin (BHI) and insulin lispro (LI) at different doses.
Both of the insulins could reduce blood glucose levels promptly after pulmonary administration. But LI showed a better tendency on hypoglycemic effect than BHI in the in vivo studies. In the in vitro studies, the apparent permeability coefficient (Papp) for BHI and LI were almost constant with increasing concentrations, which implied that insulin maybe passively diffuse through the Rana catesbeiana pulmonary membrane barrier. Interestingly, the Papp of LI was obviously higher than that of BHI, indicating that the permeability of LI across Rana catesbeiana pulmonary membrane was more effective than that of BHI.
These in vitro and in vivo results suggested that LI was easier to be absorbed in the lung than BHI and Rana catesbeiana pulmonary membrane had a potential ability, as a transport model, to predict in vivo pulmonary absorption of insulin.
[Show abstract][Hide abstract] ABSTRACT: 1. This study investigated the pharmacokinetics of thiamphenicol glycinate (TG) and thiamphenicol (TAP) in beagles (n = 6) after intravenous administration of 50 mg/kg TG hydrochloride. Plasma concentrations of TG and TAP were measured by a HPLC-UV method. 2. Two-compartment model was selected to describe the pharmacokinetic characteristics of TG and TAP in vivo. Main parameters were as follows: AUC(0-∞) of TAP and TG were 16,328 ± 1682 µg·min/mL and 3943 ± 546 µg·min/mL, respectively. The total plasma clearance (CL) of TG and TAP were 12.7 ± 2.0 mL/min/kg and 2.5 ± 0.3 mL/min/kg, respectively. Mean residence time (MRT) of TG and TAP were 27.5 ± 3.5 and 207.2 ± 20.2 min, respectively. The transformative rate constant (k(1M)) from TG to TAP was 0.0477 ± 0.0028 min(-1). The elimination rate constant (k(M10)) from TAP was 0.0238 ± 0.0044 min(-1). Coefficients of variation (CV) between observed values and predicted ones were 5.9% and 18.2%, respectively. The volume of distribution of the central compartment for TG (V(C)) and TAP (V(CM)) were 0.264 ± 0.022 L/kg and 0.127 ± 0.023 L/kg, respectively. 3. Pharmacokinetic parameters suggested that TG was presumably cleaved quickly by tissue esterase to release TAP for effectiveness in beagles after administration.
[Show abstract][Hide abstract] ABSTRACT: As a promising type 2 anti-diabetic agent, glucagon-like peptide-1 (GLP-1) is attracting more and more interest. Mutated GLP-1
(mGLP-1) is an analog of native GLP-1. To facilitate the production and purification of mGLP-1, auto-induction and on-column
cleavage was employed in this study. By using auto-induction system, after 24h of shaking culture, about 12.6g wet bacterial
cells could be obtained from 1l medium, and this was about 3.6 times more than that of the IPTG-induction group. After disruption
and centrifugation, the fusion protein was directly purified and cleaved on Ni–Sepharose 6 Fast Flow column. Then, RESOURCE15
RPC column was used for further purification. By using these two steps of purification, about 1.58mg of mGLP-1 with the purity
of up to 98% could be obtained from 1g wet bacterial cells. In the bioactivity study, mGLP-1 displayed a significant and
dose-dependent glucose-lowering activity. These results suggested that auto-induction and on-column cleavage could facilitate
the production and purification of mGLP-1. These methods could also be applied to the preparation of other proteins and peptides.
KeywordsGlucagon-like peptide-1-Auto-induction-On-column cleavage-Expression and purification
World Journal of Microbiology and Biotechnology 09/2010; 26(9):1675-1682. DOI:10.1007/s11274-010-0345-3 · 1.35 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: To facilitate expression and purification of an analog of GLP-1 (mGLP-1), an intein system was employed in this study. A recombinant fusion protein, CBD-DnaB-mGLP-1, was constructed and expressed in the form of inclusion body. After refolding, the intein-mediated self-cleavage was triggered by pH and temperature shift. By using chitin beads column followed by single step purification, about 2.58 mg of mGLP-1 with the purity of up to 98% could be obtained from 1 L medium. Tricine-SDS-PAGE, RP-HPLC, and ESI-MS were undertaken to determine the purity and molecular weight of mGLP-1. The glucose-lowering activity of mGLP-1 was also preliminarily determined.
Protein and Peptide Letters 05/2010; 17(10):1245-50. · 1.74 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: A sensitive liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) method was established and validated for the determination of glycyrrhizin in dog plasma. After treatment with methanol to precipitate proteins, plasma samples were analyzed on a reversed-phase C18 (ODS) column with a mobile phase of methanol:1% formic acid solution (75:25, v/v). MS determination was performed using negative electrospray ionization (negative ESI) in the selected ion monitoring mode. Glycyrrhizin was monitored at the m/z 821 channel and internal standard (gliquidone) at the m/z 526 channel. The calibration curve was linear over the range from 0.05 µg mL(-1) to 10 µg mL(-1) with a correlation coefficient above 0.99. This method was successfully applied to the pharmacokinetic studies in beagle dogs. The absolute bioavailability of glycyrrhizin in beagle dogs was 3.24%.
European Journal of Mass Spectrometry 01/2010; 16(6):673-8. DOI:10.1255/ejms.1101 · 1.17 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Glycyrrhetinic acid, the active metabolite of glycyrrhizin, is primarily eliminated by glucuronidation reaction in vivo. In spite of the widespread clinical use of glycyrrhizin, UDP-glucuronosyltransferase (UGT) isoforms involved in the glucuronidation of this drug are still unknown. This report identifies and characterizes the UGT isoforms responsible for glycyrrhetinic acid glucuronidation. In the enzymatic kinetic experiment performed with pooled human liver microsomes (HLMs), K(m) was 39.4 microM and V(max) was 609.2 pmol/min/mg protein. Of the baculosomes expressing 12 recombinant UGTs investigated, UGT1A1, 1A3, 2B4 and 2B7 showed catalytic activity and UGT1A3 exhibited the highest activity. K(m) values of recombinant UGT1A3 and 2B7 were 3.4 and 4.4 microM, respectively. Both imipramine (typical substrate of UGT1A3 and 1A4) and flurbiprofen (typical substrate of UGT2B7) inhibit the glucuronidation of glycyrrhetinic acid. Estimated IC(50) values were 138 microM for flurbiprofen and 207 microM for imipramine in the inhibition of the glucuronidation of glycyrrhetinic acid in HLMs. These results suggest that glycyrrhetinic acid glucuronidation is primarily mediated by UGT1A1, 1A3, 2B4 and 2B7.
Drug Metabolism and Pharmacokinetics 01/2009; 24(6):523-8. DOI:10.2133/dmpk.24.523 · 2.86 Impact Factor