4.Effect of Intracerebral Transplantation of Mesenchymal Stem Cells Derived from Human Umbilical Cord Blood on Hypoxic-Ischemic Brain Damage in Neonatal Rat
gui-zhi, XIA ; xin-ru, HONG ; xin-min, CHEN ; shui-liang, WANG ; feng-hua, LAN
Journal of Applied Clinical Pediatrics 2006;0(14):-
Objective To investigate the therapeutic effect of intracerebral transplantation of mesenchymal stem cells(MSCs) derived from human umbilical cord blood(UCB) on hypoxic-ischemic brain damage(HIBD) in neonatal rat.Methods Twenty samples of human UCB were collected from healthy full-term newborns.MSCs were isolated from human UCB by density gradient centrifugation and purified by adhere cell selection method.For transplantation,P3 human UCB-derived MSCs were labeled by the 5-bromo-2-deoxyuridine (BrdU).Thirty SD rats of 7 d were built for neonatal HIBD model.One rat died and others were divided into transplant group(n=18) and control group(n=11).At the third day after building models,human UCB-derived MSCs were injected into left cortex in transplant group,while PBS of the same volume was injected into the same site in control group at the same time.The seventh day after transplantation,6 rats of transplant group were sacrificed to prepare brain tissue sections.The survival,migration and differentiation of the transplanted cells were investigated by brain tissue immunohistochemical analysis,and nervous function of 2 groups were evaluated by modified neurological severity score(mNSS) on the first,7th,14th,21th and 28th day after transplantation.Results MSCs were isolated from 5 of 20 human UCB samples.Immunocytochemical analysis of brain tissue showed that the transplanted human UCB-derived MSCs could survive and migrate around by the center of transplant site.There were (12.67?2.73)% of MSCs differentiated into astrocyte-like cells.mNSS showed that the score of transplant group was lower than that of control group on the first,7th,14th,21th and 28th day,and the differences of score points between 2 groups on the 14th,21th and 28thday were statistically significant(Pa
5.High-Level Expression of Anti-Vascular Endothelial Growth Factor Chimeric Antibody in Eukaryotic Cells
Yu-Liang RAN ; Zhi-Hua YANG ; Li-Xin SUN ; Long YU ; Jun LIU ; Zhi-Wei DONG
Chinese Journal of Cancer 2001;20(3):282-286
Objective:This study was designed to express chimeric anti-VEGF (vascular endothelial growth factor) antibody in dihydrofolate reductase-deficient Chinese hamster ovary (CHO-dhfr-)cells at high-level, and explore an optimum method to obtain high-level expression cells clone. Methods:The light chain and heavy chain genes of chimeric anti-VEGF antibody were induced into CHO-dhfr-cells using a novel eukaryotic high-level expression vectors system for genetic engineering antibodies. High-level expression was achieved after subcloning and several rounds of co-amplification of methotrexate (MTX). Biological features and productive amount of chimeric antibody was charactered by ELISA. Result:The cells strain that secret anti-VEGF chimeric antibody at the highest level of 28 μ g/ml was established. The cells were subcloned following each round of co-amplification of MTX, while greatly different results were obtained using three methods. The chimeric antibody contained constant regions of human immunoglobin and had the specificity against VEGF by ELISA. Conclusion:The anti-VEGF mouse-human chimeric antibody was expressed at high-level successfully in CHO cells. This may be an optimum method to obtain high-level expression cells clone for the eukaryotic high-level expression vectors system.
6.DEGRADATION AND TRANSFORMATION OF COTTON SEED HULLS BY COPRINUS COMATUS
Xin-Jiang NI ; Zhi-Yong FENG ; Li-Kun LIANG ; Cui-Rong YOU ; Ying-Jie PAN ;
Microbiology 1992;0(02):-
Coprinus comatus cultivated on cotton seed hull medium decomposed lignocellulose straggly and was high of absolute biological efficiency. Lignocellulose is the main carbon source for the fruiting stage of the fungus. There existed the positive correlation between the degradation rates of the cellulose and hemicellulose in the medium and the activities of extracellular CMCase (carboxymenthelcel lulase), FPase (filter paper cellulase) and HCase (hemicellu-lase), there also existed the positive correlation between the degradation rate of the lignin in the medium and the activity of extracellular laccase, but no correlation between the degradatio rate of the lignin in the medium and the activity of peroxidase. The activity of extracellular amylase was comparatively high at mycelial growth stage, and the protease activity peek was at teh time when the fruitbody matured.
7.CLONING OF NATTOKINASE GENE AND EXPRESSION IN E. COLI
Li-Xin LUO ; Zhi-Li HUANG ; Ru-De YANG ; Jun-Jian LING ; Shi-Zhong LIANG ;
Microbiology 1992;0(03):-
In this study, nattokinase gene was amplified by PCR using bacillus subtilis chromosomal DNA as template and cloned into expressed vector pBV220. After transforming recombinant plasmid into E.coli HB101, the recombinant strain was yielded. It was proved that expression products was secretive and expression protein was 12% of total cell protein by SDS-PAGE. Optimum culture time and inducing time was determined as 6h and 5h respectively. The plasmid stability studies showed that recombinant plasmid has excellent segregational stability but the structural stability was not good in the host cell.
8.The clinical observation of oxaliplatin combined with capecitabin for patients with advanced colorectal cancer
Xin-Jian RUAN ; Jian-Fei WANG ; Zhi GUO ; Zhao-Liang CHEN ; Xia ZHANG ;
Cancer Research and Clinic 2000;0(06):-
Objective To observe the therapeutic effect and toxicity reaction of Oxaliplatin com- bined with XeLoda in the treatment of 42 patients with advanced colorectal cancer.Methods All the pa- tients were treated with Oxaliplatin(130 mg/m~2,ivgtt for 2 h,d1)combined with XeLoda(2000 mg?m~(-2)?d~(-1), po,bid,d1 to d14).The regime was repeated every 21 days for at least 3 consecutive cycles.Results The to- tal response rate was 40.5%(17/42)in which 2 got CR and 15 PR.24 patients got Kamofsky score increased. The major toxic effects were alopecia,peripheral neuritis,gastrointestinal tract reactions and myelosuppression. Conclusion Oxaliplatin combined with XeLoda regimen is effective in the treatment of advanced colorectal cancer,and its toxicity is tolerable.It is worth studying in the future.
9.Effects of isorhamnetin on human liver microsomes CYPs and rat primary hepatocytes
Rongjia LIANG ; Junxiu CHEN ; Dexian ZHI ; Yaowen FAN ; Wenli LIU ; Xin HE
Drug Evaluation Research 2017;40(5):627-632
Objective To study the inhibitory effects ofisorhamnetin on six kinds of CYPs of liver in vitro,and the toxic effect on rat hepatocytes Methods This report uses warm incubation of human liver microsomes in vitro to investigate the inhibition of isorhamnetin on 6 kinds of CYPs (CYP2C19,CYP2D6,CYP3A4,CYP2E1,CYP1A2 and CYP2C9),and using HPLC-MS/MS to detect product of metabolism as well as analysing of the pathways of metabolic.At the same time,using rat primary hepatocytes which has low CYPs activity in vitro to explore whether the use of isorhamnetin will cause effects on the ALT,AST and LDH of hepatocytes.Results Isorhamnetin has inhibition effects on CYP2E1 and CYP1A2,the inhibition rate were 59.48% and 39.91%,respectively.Methylated metabolite is produced after incubating of isorhamnetin and HLMs.The isorhmnetin becomes high polarity and water solubility metabolite 3,3',4',5,7-hydroxyflavone.Isorhamnetin of 30,100 and 300 μmol/L cause a significant rise of ALT and LDH in primary cultured rat hepatocytes cultured (P < 0.01).isorharnnetin of 100 μmol/L cause a rise of AST in primary cultured rat hepatocytes cultured (P < 0.05) and 300 μmol/L cause a significant rise (P < 0.01).It was a dose-dependent manner.Conclusion Isorhamnetin in vitro mainly metabolized by HLMs,and at the same time have a certain inhibitory effect on CYP2E1 and CYP1A2,which may cause the drugs which are metabolized by CYP2E1 and CYP1A2 in vivo accumulation that lead to a series of drug interactions.The results also indicate that heavy use of isorhamnetin cause some adverse effects on hepatocytes,and it was a dose-dependent manner.Individuals need to pay attention to the dose ofisorhamnetin and the potential drug interactions.
10.Research progress of natural products for the treatment of hyperuricemia and gout
Zhi-jiao ZHANG ; Rui-peng LIANG ; Tong ZHAO ; Shu-jing XU ; Xin-yong LIU ; Peng ZHAN
Acta Pharmaceutica Sinica 2022;57(6):1679-1688
Although current synthetic anti-gout drugs have significant therapeutic effects in reducing serum uric acid levels, they have serious side effects such as allergic reactions and liver and kidney damage. Natural products with a wide range of uric acid-lowering and high safety have played a critical role in anti-gout drug discovery and development. This paper reviews the natural products with uric acid-lowering or anti-gout pharmacological effects and the investigation on their mechanisms of action, to provide information for drug discovery and development.