1.C/EBPα is involved in transcriptional regulation of mVGLUT2 gene expression
Zhuguo WU ; Shiming LIU ; Xinchun CHEN
Chinese Journal of Pathophysiology 2010;26(1):142-145
AIM: The main purpose of this study is to investigate the regulatory role of C/EBPα in mouse vesicular glutamate transporter 2(mVGLUT2) gene expression. METHODS: The promoter region of mVGLUT2 was cloned to PGL3-basic vector. Site-direction mutation was used to identify the CCAAT-enhancer-binding protein α(C/EBPα) binding site. The promoter activity was observed by luciferase system. The binding between C/EBPα protein and mVGLTU2 promoter region was determined by EMSA. The C/EBPα gene expression was inhibited by its specific siRNA. RESULTS: mVGLUT2 promoter activity decreased about 50% after mutation of C/EBPα binding site. EMSA showed that C/EBPα protein bound onto mVGLUT2 promoter region. Meanwhile, when C/EBPα gene expression was inhibited by its specific siRNA, mVGLUT2 promoter activity, mRNA level and protein level were decreased about 60%, 40% and 45%, respectively. CONCLUSION: C/EBPα is involved in the regulation of mVGLUT2 gene expression.
2.Effects of TGF-?_(1,2) on basal and FSH-stimulated production of estradiol and progesterone by human granulosa cells in vitro
Shiming LV ; Hong LV ; Yuanyue CHEN ;
Chinese Journal of Laboratory Medicine 2001;0(05):-
Objective To investigate the possibility that TGF ? may also influence the basal and FSH stimulated production of estradiol and progesterone by granulosa cells in vitro. Methods Granulosa cells were obtained from infertile patients undergoing in vitro fertilization and embryo transfer treatment and cultured with serum free supplemented HAM′s F 10 medium(control) or increasing concentrations of TGF ? 1,2 in the absence or presence of FSH (96 U/ml). The media were collected in 24, 48, 72 and 96 h intervals and assayed for estradiol and progesterone. Results Increasing amounts of TGF ? had a significant effect on estradiol production, especially in the presence of FSH,The estradiol value was evaluated from (12 077.9?5 835.3) pmol/L and (13 656.1? 8 404 3) pmol/L to (16 606.8?7 993.3) pmol/L and (19 480.4?913.8) pmol/L. TGF ? inhibited FSH stimulated progesterone but not unstimulated progesterone release ( P
3.Assessment of multiple quantitative fluorescence PCR technique for common chromosome ;aneuploidies
Yan CHEN ; Yuning ZHU ; Shiming LYU
Chinese Journal of Laboratory Medicine 2017;40(1):50-54
Objective To evaluate the effectiveness of multiple quantitative fluorescence PCR ( QF-PCR) as a rapid technique for prenatal diagnosis of common chromosome aneuploidies , in order to optimize the prenatal diagnosis and shorten the period of diagnosis.Methods Totally 731 amniotic fluid samples of pregnant subjects ,who were referred to the Women′s Hospital School of Medicine Zhejiang University during August 2013 and September 2015, were analyzed with conventional karyotype and the QF-PCR technique by short tandem repeat(STR) markers to detect chromosomes 13,18,21,X and Y aneuploidies.There were 558 samples detected by single blind method , 173 samples detected by double blind method.Results All of the 731 amniotic fluid samples were tested in this study by QF-PCR and the results were compared to the conventional cytogenetic analysis results of the same sample.Totally 558 samples with single blind method detected 5 trisomy 21, 2 trisomy 18, 1 trisomy 13, 1(45,X), 1(47,XXY), 1(47,XYY), 1(47,XXX) and 1(69,XXX), 173 samples with double blind method detected 1 trisomy 21 and 1 trisomy 18.The rapid QF-PCR assay was successful to detect all aneuploidies involving chromosomes 21, 18, 13, X and Y in prenatal diagnosis , which were verified by chromosome karyotype analysis.The results of QF-PCR method were compared with the results of chromosome karyotype analysis , the positive rate was 15/16, the negative rate was 100%(715/715).Non chimeric chromosome abnormality detection rate was 15/15.Conclusions The multiple QF-PCR was a reliable method of detecting common chromosome aneuploidies for rapid prenatal diagnosis.As an important supplement of karyotype analysis , it was of great significance to optimize and improve the prenatal diagnosis system , and might provide more appropriate diagnostic methods for pregnant women.
4.Clinicopathological features of central nervous system hemangioblastoma in von Hippel-Lindau syndrom
Weidong MAO ; Qi HUANG ; Shiming CHEN
Journal of Regional Anatomy and Operative Surgery 2016;25(11):848-850
Objective To investigate the clinicopathological features of central nervous system hemangioblastoma in von Hippel-Lindau syndrome.Methods The data of 10 patients with central nervous system hemangioblastoma in our hospital since 2013 were analyzed retro-spectively,and the clinicopathological features of central nervous system hemangioblastoma in von Hippel-Lindau syndrome were summarized. Results The macroexamination result showed that most tumor lesions were found in the cerebellum and medulla oblongata,with cystic chan-ges,size from 1 cm to 5 cm,the average size was (3.1 ±0.2)cm,clear boundary,intracapsular yellow cyst fluid.The microscopy result showed tumor foci with a rich blood supply,endothelial cell proliferation and hypertrophy in vascular,which arranged in nests or lobulated mesenchymal cells,the cytoplasm of stromal cells was abundant and lightly stained,a rich lipid was seen,with vesicular or foam.Conclusion The von Hippel-Lindau syndrome is usually cystic lesion,the microscopic examination shows tumor foci with rich blood supply,endothelial cell proliferation,hypertrophy and other changes in vascular.
5.Neoadjuvant Chemotherapy for Potentially Resectable Gastric Cancer
Shiming XIAO ; Chen WU ; Huaiwu JIANG
Chinese Journal of Bases and Clinics in General Surgery 2008;0(08):-
Objective To summarize the current value of neoadjuvant chemotherapy(NAC) for potentially resectable gastric cancer.Methods The recent 5-year literatures searched through the PubMed with the key words:stomach neoplasm,gastric cancer/carcinoma,neoadjuvant therapy/chemotherapy and preoperative therapy/chemotherapy as well as the relevant reports presented in the ASCO Annual Meeting in 2007 and 2008 were analyzed.The present status of NAC for advanced gastric cancer was summarized,the necessity and feasibility were evaluated,and the patients features for selecting,the predictors for response,the mainly existing problems and development trend of NAC were analyzed.Results At present,there were 7 randomized control trails(RCT) published,and among them 3 were phase Ⅲ.It was safe,effective and feasible to most of trails in NAC for gastric cancer.However,it was still little to obtain survival benefit for NAC RCT,and short of randomized trial comparing strict preoperative chemotherapy to surgery alone or perioperative chemotherapy to surgery plus adjuvant chemotherapy.It remained lots of problems such as how to select the appropriate patients,the effective induced regimes and the predicted factors,the evaluated indices for response.Conclusion NAC is a safe,feasible and efficient method to potentially resectable gastric cancer,but strict phase Ⅲ randomized trials are needed.In the future,substantial improvements of treatment outcome will likely depend on the novel drugs and molecular biological targeted therapies.
6.Construction of evaluation index system for postgraduate tutors in medical universities
Ye CHEN ; Shiming LIU ; Mingsong XU
Chinese Journal of Medical Education Research 2013;(1):2-5
Objective To build a scientific,rational and practical evaluation index system for medical tutors.Methods Evaluation framework was initially constructed on the basis of literature analysis.Two rounds questionnaires investigation were conducted soon using Delphi method and the data were analyzed using descriptive analysis,independent samples of non-parametric test (Kreskas-Wallis test),expert reliability analysis,analytic hierarchy process and other methods.Results The results of two rounds of expert questionnaires showed that both recovery rate and acceptance rate of the questionnaires were more than 90% and overall expert coefficient was 0.8957.The coordinate coefficient of the second-class index was 0.181,x2 =27.148,P < 0.001,and that of the third-class index was 0.157,x2 =127.03,P < 0.001.Experts' opinions were highly consistent.Conclusions Evaluation index system including three first-class indexes,seven second-class indexes and twenty-eight third-class indexes was established for medical tutors.The evaluation index system is of great importance in safeguarding postgraduate culturing quality and strengthening teaching staff construction.
7.C/EBP? is involved in transcriptional regulation of mVGLUT2 gene expression
Zhuguo WU ; Shiming LIU ; Xinchun CHEN
Chinese Journal of Pathophysiology 1986;0(01):-
AIM:The main purpose of this study is to investigate the regulatory role of C/EBP? in mouse vesicular glutamate transporter 2(mVGLUT2) gene expression. METHODS:The promoter region of mVGLUT2 was cloned to PGL3-basic vector. Site-direction mutation was used to identify the CCAAT-enhancer-binding protein ?(C/EBP?) binding site. The promoter activity was observed by luciferase system. The binding between C/EBP? protein and mVGLTU2 promoter region was determined by EMSA. The C/EBP? gene expression was inhibited by its specific siRNA. RESULTS:mVGLUT2 promoter activity decreased about 50% after mutation of C/EBP? binding site. EMSA showed that C/EBP? protein bound onto mVGLUT2 promoter region. Meanwhile,when C/EBP? gene expression was inhibited by its specific siRNA,mVGLUT2 promoter activity,mRNA level and protein level were decreased about 60%,40% and 45%,respectively. CONCLUSION:C/EBP? is involved in the regulation of mVGLUT2 gene expression.
9.Simultaneous Determination of Contents of Piperacillin and Tazobactam in a Preparation With RP-HPLC
Zeming YANG ; Jisheng CHEN ; Shiming YU ; Zhuoxia CHEN
China Pharmacy 1991;0(01):-
OBJECTIVE:To establish a RP-HPLC method to simultaneously determine the contents of piperacillin and taz_obactam METHODS:Using ? Bondapak C18 column as fixed phase,a mixture of methanol and purified water(34 5∶65 5)as mobile phase,and caffeine as internal standard,detecting wavelength:220nm,flow rate:1 0ml/min RESULTS:The linear rangs of piperacillin and tazobactam were 40~200?g/ml and 5 0~25 0?g/ml respectively The average recovery and RSD (n=5) were (100 2?0 61)% for piperacillin and (99 03?0 90)% for tazobactam CONCLUSION:This method is rapid,simple and accurate
10.Establishment and Application of 18-STR Database in Han Population from Zhejiang Province
Yan CHEN ; Yuning ZHU ; Shiming LU ; Jianfei YOU ; Yu MA
Journal of Sun Yat-sen University(Medical Sciences) 2010;31(1):122-128
[Objective] To construct a database for the genetic polymorphism of 18 STR loci (D8S1179, D21S11, D7S820, CSFIPO, D3S1358, TH01, D13S317, D16S539, D2S1338, D19S433, VWA, TPOX, D18S51, D5S818, FGA, PentaE, PentaD, SE33) in Hart population from Zhejiang province. To investigate the application of 18 STR loci in the field of paternity testing and prenatal diagnosis. [Methods] Fluorescent dye labeling multiplex STR-PCR, capillary electrophoresis and DNA sequencer GeneScan were adopted in genotyping 598 unrelated samples collected from Han population in Zhejiang province. 18-STR database was established and analyzed. Population comparison was conducted between Han population in Zhejiang province and 8 other population. 15-STR and 18-STR identification system were compared in 497 paternity testing cases. [Results] We observed the distribution of 18 STR loci in Han population meet Hardy-Weinberge equilibrium and was different from other 8 population (X~2 test, P>0.05). Statistical results showed that the heterozygosis (He) ranged from 0.630 to 0.942. The combined power of discrimination was>0.9999999999. Compared with 15-STR identification system, higher paternity index scores and higher exclusion rate were obtained with 18-STR identification system in dual-case paternity test and mutation identification. One trisomy 21 fetus was found in a prenatal paternity test case which had two characteristic genotypes in 2 STR loci of D21S11 and Penta D. [Conclusions] The 18 loci were relatively highly genetic polymorphic in Zhejiang Han population and could be used for paternity testing. Some STR loci could be used in prenatal diagnosis for aneuploidy.