1.Eukaryotic expression and biological activities of anti-p185~(erbB2) mouse/human chimeric antibody
Beihai JIANG ; Wenbin LIU ; Lin MENG ; Jian WU ; Chengchao SHOU
Journal of Peking University(Health Sciences) 2003;0(05):-
Objective: Overexpression of the HER2/neu oncogene is a frequent molecular event in multiple human cancers. Being a cancer antigen, p185 erbB2 is an ideal target for immunotherapy. In order to decrease the immunogenicity of mouse anti-p185 erbB2 monoclonal antibody in human cancer therapy, we constructed the eukaryotic expression vector of anti-p185 erbB2 chimeric monoclonal antibody and verified expression of the chimeric antibody in CHO-dhfr - cell. Methods: The variable regions of light chain and heavy chain were amplified with RT-PCR and inserted into the chimeric antibody vector pWSD2. After CHO-dhfr - cells were transfected with recombination plasmid by lipofectAMINE, the chimeric antibody expressing level was identified with RT-PCR, indirect-ELISA, and Western blot. The specificity of the anti-p185 erbB2 chimeric antibody was testified with ELISA assay and immunoprecipitation. Moreover, the effects of chimeric antibody on the proliferation of breast cancer cell line SKBR3, which is overexpressing p185 erbB2 , were measured with MTT assay in vitro. Results: The anti-p185 erbB2 chimeric antibody eukaryotic expression vector was constructed successfully and the expression of the chimeric antibody in CHO-dhfr - was verified by RT-PCR, indirect-ELISA, and Western blot. ELISA assay showed that chimeric antibody reacted with cells overexpressing p185 erbB2 specifically, but did not react with that non-overexpressing p185 erbB2 . Immunoprecipitation test confirmed that the chimeric antibody could bind to p185 erbB2 specifically. The MTT assay demonstrated that the chimeric antibody could inhibit the growth of SKBR3 cells overexpressing p185 erbB2 . Conclusion: The anti-p185 erbB2 mouse/human chimeric antibody that was expressed in CHO-dhfr - cells can bind to p185 erbB2 specifically and inhibit proliferation of SKBR3 cells overexpressing p185 erbB2 . It has a potential application in biotherapy of cancer.
2.Small caliber vascular tissue engineering progress
Xiaolong MA ; Wenbin LI ; Diankun LI ; Zhifei XIN ; Xu MENG
Chinese Journal of Thoracic and Cardiovascular Surgery 2016;32(6):374-376,384
Review preparations about small caliber artificial blood vessels,cell cultivation,nano materials and growth factors in the research of vascular tissue engineering.The main method of keeping small diameter vascular graft patency is to promote the small-diameter artificial vascular cells,using nanometer encapsulated growth factor delivery system which can promote the growth of endothelial cells at the luminal surface adhesion,and using anticoagulant drug delivery system.
3.miR-126 promotes endothelial progenitor cell migration and targets KANK2
Qingyou MENG ; Wenbin WANG ; Zhixin CAI ; Bin SHANG ; Xiaoqiang LI
Chinese Journal of General Surgery 2013;28(8):611-614
Objective To investigate the role of miR-126 (micro RNA-126) in rat endothelial progenitor cells (EPCs) proliferation and migration and the starget gene of miR-126 by bioinformatics and experimental survey.Method EPCs were transfected with control oligoes and miR-126 mimics or inhibitor by electroporation.MTT was performed to evaluate the growth of EPCs subjecting to miR-126 overexpression.Cell migration analysis was done by wound healing and transwell assay.The target genes of miR-126 were predicted by TargetScan and validated by Western blot.Result (1) miR-126 mimics promoted EPCs growth at 24 h post cell transfection (P < 0.01).In contrast,the EPCs growth was immue from miR-126 application at 48 and 72 h.(2) Both the wound healing and transwell assay show that miR-126 promotes EPCs migration (P < 0.01) and miR-126 inhibitor inhibits EPCs migration (P < 0.01).(3)It is predicted that KANK2 is the potential target gene of miR-126 by TargetScan online software.(4) The results of Western blot indicated that miR-126 mimics repress the expression of KANK2 compared with NC but miR-126 inhibitor enhances KANK2 expression.Conclusions miR-126 has a transient effect on the promotion of EPCc growth.miR-126 promotes EPCs migration and targets KANK2 protein.
4.Diagnostic value of HbA1C and fasting plasma glucose as early screening parameters for diabetes mellitus
Chunfang YU ; Jun GE ; Jingyi LI ; Jian MENG ; Wenbin LIU ; Junjun LIU
Chinese Journal of Endocrinology and Metabolism 2011;27(5):390-392
The validity of HbA1C and fasting plasma glucose(FPG)for screening of diabetes in community population was investigated.A total of 1 794 subjects from two sub-districts of Yangpu District,Shanghai,underwent a 75 goral glucose tolerance test(OGTT)and HbA1C determination.Based on 1999 World Health Organization criteria,there were 1 411 subjects with normal glucose tolerance(NGT),111 impaired fasting glucose(IFG),73 impaired glucose tolerance(IGT),33 IFG+IGT,and 166 diabetes.According to the receiver operating characteristic curve,the optimal cut-point of FPG for diagnosing diabetes was 6.15 mmol/L with sensitivity of 89.0%,specificity of 92.8%,and area under the curve of 0.959.The cut-point of HbA1C for diagnosing diabetes was 6.5% with a sensitivity and specificity of 71.1% and 80.2%.The area under the curve was 0.822.The screening model using FPG ≥6.1mmol/L and HbA1C≥6.5% had sensitivity of 66.9% and specificity of 97.0%.When the model was FPG ≥6.1 mmol/L or HbA1C ≥6.5%,the sensitivity and specificity was 96.3% and 76.7% respectively.The results suggest that both FPG and HbA1C have good value for screening diabetes in community,and FPG combined with HbA1C may further promote diagnostic efficacy.
5.Effects of Triptolide on Proliferation and Apoptosis of Cyst-lining Epithelial Cells of Polycystic Kidney Disase
Huanrong YANG ; Shumei YANG ; Guangxin WANG ; Fanjie MENG ; Shufang CAI ; Wenbin WU ; Shijun WANG
Herald of Medicine 2016;35(9):920-924
Objective To investigate the effect of triptolide (TP) on proliferation and apoptosis of cyst-lining epithelial cells with autosomal dominant polycystic kidney disease ( ADPKD). Methods Primary cultured cyst-lining epithelial cells were treated with TP at different concentrations for 12 h,24 h,48 h and 72 h, respectively.The proliferation activity of the cells was evaluated by Brdu assay. The cell cycle distribution was determined by flow cytometry. The apoptotic and apoptotic ratio were determined by FITC-AnnexinV binding/ PI. The morphological changes of cyst-lining epithelial cells were observed under transmission electron microscope. Results TP significantly inhibited the proliferation of cyst-lining epithelial cells and induced apoptosis in a dose- (10-40 ng?mL-1 )and time-dependent(12-48 h) manner. Typical ultrastructural changes of apoptotic cells were observed under electron microscope. Conclusion TP significantly inhibited the proliferation of cyst-lining epithelial cells and induced the apoptosis of cyst-lining epithelial cells, thus inhibited cyst forming and delayed cyst developing. The mechanism may involve several targets and pathways.
6.Construction of vascular sustained release model of vascular endothelial growth factor
Xiaolong MA ; Wenbin LI ; Zhifei XIN ; Diankun LI ; Zifan ZHOU ; Juyi WAN ; Xu MENG
Chinese Journal of Thoracic and Cardiovascular Surgery 2016;32(10):625-628
Objective To construct a model of small caliber vascular endothelial growth factor(VEGF) in tissue engineering,to investigate the performance of the sustained-release microspheres and vascular stent,and to provide materials and theoretical basis for animal experiment.Methods The sheep carotid arteries were treated with a cellular reagents,the cellular conditions and the stent properties were observed.Preparation of sustained release microspheres containing VEGF,particle size,encapsulation efficiency,drug loading and release curve were measured.The effective combination of the slow release microsphere and the vascular stent was used in the freeze drying technology.The rat vascular endothelial cells grown in tissue engineered blood vessel model release lumen,observe the growth of endothelial cells.Results After the treatment,the original performance of the vascular stent can be maintained.The average particle size of the microspheres was (9.8 ± 6.0) μm,which could be released slowly in 20 days,and the release rate was 70%.Microspheres can effectively with the tissue.engineering blood vessel tight binding.Rat vascular endothelial cells can grow in the vascular stent surface.Conclusion Using Triton X-100,DNA/RNA ribozyme for acellular reagent,stent performance is good.PLGA microspheres have good sustained release performance,and constructing appropriate tissue engineered small caliber vascular release model by using freeze drying technology can make the stent compact structure.
7.Technical details of ultrasound guided mammotome minimally invasive biopsy system for benign breast mass in 186 cases
Wenbin GUO ; Baochen SUN ; Qingru MENG ; Wei GAO ; Shuke GE ; Linlin GAN
Chinese Journal of General Surgery 2008;23(6):438-440
Objective To explore the clinical results and operation experiences for benign breast mass by ultrasound guided mammotome minimally invasive biopsy system (MMIBS). Methods 212 benign breast masses in 186 patients were resected by ultrasound guided MMIBS. Clinical data of 186 patients were retrospectively analyzed. Results Needle position in 186 patients was visualized. Lesions were completely removed in 134 cases of 186 (72%) patients. The complete resection rate for tumors on major pectoral muscle or near areola were 31.5% (6/19) and 33.3% (4/12) respectively. Identified by postoperative ultrasound, 118 out of 134 patients (88.0%) with tumor sizes 0.5 to 2.5 cm and 16 out of 38 patients (42.1% ) with sizes 2.5 to 3.0 cm were completely removed. No lesions larger than 3.0 cm were completely removed. All 52 cases in which the tumors were not completely removed by MMIBS were converted to open surgery. Ultrasound follow-up after 4 weeks showed that all the 134 cases that had had masses completely removed had no residual masses, whereas 6 months after operation, 16 out of the 112 cases proved tumor recurrent necessitating open reoperation in 6 cases and second MMIBS operation in 10 cases, among them one case recurred after six months and received open operation. Conclusions For small benign breast mass, MMIBS has therapeutic effect with significantly minimal invasion.
8.64-slice CT angiography in lower extremity peripheral arterial occlusive disease: clinical value of test injection at popliteal arteries
Zheng SHU ; Xiaofei DENG ; Chenjin GE ; Feng SUN ; Yinge ZOU ; Wenbin MENG
Chinese Journal of Radiology 2011;45(7):674-677
Objective To investigate the clinical value of the test injection at popliteal arteries on 64-slice CTA in lower extremity peripheral arterial occlusive disease (PAOD). Methods Twenty-eight patients with PAOD referred for 64-slice CTA were enrolled consecutively in the study. Test injection was performed at bilateral popliteal arteries (the level of knee joints) and the clinic value of the peak value and the time to peak was analyzed. The relationship between the time to peak and the peak value was evaluated with Pearson test. The time to peak was used for programming of the CT angiographic acquisitions with fast scanning mode. The quality of visualization of each arterial segment was determined independently by two radiologists. Results Fifty-four (96%, 54/56) time-attenuation curves were obtained in 28 patients. The wide interindividual variation in the peak value and the time to peak was observed in 52 curves of 26 patients with range of 60-178 HU,21-46 s and an average of (135±28) HU,(31±6) s, respectively. The difference in the peak value and the time to peak between bilateral popliteal arteries was also observed with range of 10-80 HU and an average of (32±18) HU in 19 patients,with range of 1-12 s and an average of (5±3) s in 21 patients. There was negative relationship between the peak value and the time to peak (r=-0.526, P<0.01). The CTA images were of good (598 segments) or medium quality (12 segments) in 99% segments (610/616). Conclusions The test injection at popliteal arteries was useful for 64-slice CTA in the patients with PAOD, as it could accurately specify the delay time of CT angiographic acquisitions.
9.Application of scenario simulation in prevention of needlestick injuries based on Kirkpatrick model
Lei SONG ; Wenbin JIANG ; Xinzhi SHAN ; Silong GAO ; Hui LIN ; Kun ZHANG ; Meng NIU ; Song HU
Chinese Journal of Medical Education Research 2021;20(4):478-484
Objective:To evaluate the application method and effect of standardized scenario simulation teaching based on Kirkpatrick model in vocational protection education for nursing students.Methods:A historical controlled trial study was designed. Practical nursing students enrolled in 2018-2019 and 2017-2018 were selected into the experimental group ( n=203) and control group ( n=196), respectively. The experimental group adopted standardized scenario simulation teaching in the prevention and control education of needlestick injuries, and the control group adopted traditional classroom lecture. Using the Kirkpatrick model, the teaching effect of needlestick injuries protection for nursing students were compared between the 2 groups from such 4 levels as in reaction level, learning level, behavior level and results level. Results:There was no significant difference in the baseline data between the two groups in terms of age, gender, educational background and test scores of nursing professional knowledge as compared to that before practice. In reaction level: the nursing students' satisfaction of experimental group in teaching methods ( t=25.149, P<0.001) and teaching environment ( t=12.827, P<0.001) are higher than that of the control group, with statistical significance. In learning level: the test scores of needlestick injury knowledge in experimental group were significantly higher than those in control group ( t=8.221, P<0.001). In behavior level: the level of needlestick injury protection behavior in the experimental group was significantly higher than that in the control group ( t=9.250, P<0.001), and the knowledge conversion rate in the experimental group was higher than that in the control group ( t=6.054, P<0.001). In results level: the needlestick injuries incidence of experimental group was significantly lower than that of the control group ( χ2=15.815, P<0.001), the reported rate of needlestick injuries of experimental group was significantly lower than that of the control group ( χ2=14.185, P<0.001). Conclusion:The implementation of standardized scenario simulation teaching can effectively improve the effectiveness of vocational protection learning and reduce the incidence of needlestick injuries.
10.Value of CT lymphangiography combined with direct lymphangiography in diagnosing primary intestinal lymphangiectasia
Jian DONG ; Wenbin SHEN ; Jianfeng XIN ; Meng HUO ; Chunyan ZHANG ; Pengfei LIU ; Tingguo WEN ; Rengui WANG ; Xiaobai CHEN
Chinese Journal of Radiology 2017;51(5):362-365
Objective To investigate the clinical value of CT lymphangiography (CTL) combined with direct lymphangiography (DLG) in primary intestinal lymphangiectasia (PIL). Methods Sixteen patients diagnosed as PIL with intestinal enteroscopy were recruited in this retrospective study. All the patients were performed DLG and CTL one week before exploratory laparotomy. Subjective assessment in DLG included weak lymphatic fluid drainage, lymphangiectasia, lymphatic reflux, fistula and thoracic outlet reflux or obstruction. While for CTL combined with DLG, the intestinal and extra-intestinal lesions were evaluated, including lymph node, edema, lymphangiectasia and abnormal distribution, fistula, and lymphangiomatosis. All the diagnosis was compared with intestinal endoscopy results. Results For DLG, 16 weak lymphatic fluid drainages, 9 lymphangiectasia, 1 fistula with abdomen and 14 thoracic outlets weak lymphatic fluid drainage or obstruction were found. For DLG combined with CTL, 16 intestinal lumens dilatation and 14 circumferential intestinal thickening were found in intestinal lesions. While for extra-intestinal lesions, the imaging features included edematous findings (12 in mesentery, 7 ascites only, 2 hydrothorax and ascites, and 3 pericardial, thoracic and abdominal effusions), abdominal lymph nodes (6 cases), lymphangiectasia and abnormal distributions (14 cases), fistulas (lymph-intestinal luminal fistula in 4 cases, and lymph-abdominal fistula in 3 cases), lymphangiomatosis (3 cases), and thoracic duct outlet dysfunction and reflux (14 cases).The number of cases diagnosed as intestinal lymphangiectasia, intestinal luminal lymph exudation and lymph fistula were 16, 10 and 6 with intestinal endoscopy, while the number were 11, 0, and 4 with CTL combination with DLG. Conclusion Combination of CTL with DLG is valuable in the diagnosis of PIL.