Objective:To prepare and identify monoclonal antibody specifically targeting epidermal growth factor receptor(EGFR) and(or) epidermal growth factor receptor vⅢ(EGFRvⅢ),and to investigate its inhibitory effects on human hepatocellular carcinoma Huh7-EGFRvⅢ cell-and epidermal carcinoma A431 cell-implanted tumors in nude mice.Methods: BALB/c mice were immunized with 3T3 cells stably transfected with EGFRvⅢ(3T3-EGFRvⅢ).Immunized spleen cells were fused with myeloma SP2/0 cells,and anti-EGFRvⅢ monoclonal antibody positive hybridoma cells(named 9B9 antibody and 9B9 cells,respectively) were selected and identified by ELISA.The specific interaction between 9B9 antibody and EGFRvⅢ/EGFR antigen was detected by Western blotting and immunofluorescence assay.Huh7-EGFRvⅢ cell-(human hepatocellular carcinoma Huh7 cells stably transfected with EGFRvⅢ) and epidermal cell carcinoma A431 cell-bearing mouse models were established and were divided into PBS group,Cetuximab group and 9B9 antibody group.Then,anti-tumor effect of 9B9 antibody was examined and compared with those of PBS and Cetuximab.Results: A monoclonal antibody,named 9B9 antibody,was obtained by hybridoma technique and it reacted with both EGFRvⅢ antigen and EGFR antigen as detected by Western blotting and immunofluorescence.The inhibitory rates of Cetuximab and 9B9 antibody against Huh7-EGFRvⅢ cells-implanted tumors were 42% and 46%,respectively,and those against A431 cells-implanted tumors were 85% and 86%,respectively.Conclusion: 9B9 monoclonal antibody can effectively inhibit the growth of human hepatocellular carcinoma cell-and epidermal cell carcinoma cell-implanted tumors,and the effects resemble that of Cetuximab.

Objective To explore the value of primary and Advanced levels of laparoscopic simulating training in different seniority of gynecologic residents.Methods 77 residents in their first to forth training-year were divided into two groups:1-2 year resident and 3-4 year resident,trained with different levels of simulating training plans respectively and then assessed in the Department of Obstetrics and Gynecology in Peking University Third Hospital.Results The qualified rate of primary and advanced simulation training was 85.7％ and 57.1％ respectively.The qualified rate of primary training (80.4％ vs.100.0％,P=0.028) and advanced training (12.5％ vs.57.14％,P=0.000) were significantly different between 1-2 year residents and 3-4 year residents.The operative skills improved significantly in all the residents.In the 1-2 year residents,the scores of the primary training increased more obviously,while in the 3-4 year residents,the scores of the advanced training increased significantly.Conclusion It might be more effective for residents with different seniority to receive different levels of simulating training accordingly,so as to improve their laparoscopic operative skills more effectively.

Objective:To explore the effect of Zhisanzhen acupuncture on cognition ability and apoptosis in the hippocampus using a murine model of cerebral ischemia.Methods:Thirty-two male C57 mice were randomly divided into a control group, a model group, a sham acupuncture group and a Zhisanzhen group, each of 8. All except the control group received middle cerebral artery occlusion and reperfusion (MCAO/R) surgery. Successful modeling was confirmed using Bederson scoring and 2, 3, 5-triphenyltetrazolium chloride staining. Beginning 24 hours after successful modeling, the Zhisanzhen group was given 30 minutes of acupuncture at the Shenting and bilateral Benshen points with the injection 3mm deep daily for five days The sham acupuncture group was given acupuncture at 5 mm next to the Shenting and bilateral Benshen points 3mm deep. Twenty-four hours after the last session of acupuncture, the rats′ cognition was tested using a water maze. After the experiment the hippocampus was sampled on the affected side and any apoptosis of hippocampal neurons was detected using the terminal deoxynucleotidyl transferase nick-end labeling (TUNEL) assays. Changes in caspase-3, cleaved caspase-3 protein and p38 mitogen-activated protein kinase (MAPK) protein in the hippocampal tissue were evaluated using western blotting. The total serine phosphorylation level of caspase-3 protein was detected using immunoprecipitation.Results:Compared to the model and the sham groups, the average escape latency in the Zhisanzhen group was significantly shorter and the times crossing the platform was significantly greater. There were significantly fewer TUNEL-positive cells in the Zhisanzhen group on average, and less caspase-3 and cleaved caspase-3, but significantly more p38-MAPK and greater serine phosphorylation of its substrate caspase-3.Conclusion:Zhisanzhen acupuncture can ameliorate recognition difficulties after cerebral ischemia. It promotes phosphorylation of caspase-3, limiting its cleavage and activation, and decreasing neuron apoptosis in the hippocampus.

Cholesterol-dependent cytolysins (CDC) are pore forming toxins. A prototype of the CDC family members is perfringolysin O (PFO), which directly binds to the cell membrane enriched in cholesterol, causing cell lysis. However, an exception of this general observation is intermedilysin (ILY) of Streptococcus intermedius, which requires human CD59 as a receptor in addition to cholesterol for its hemolytic activity. A possible explanation of this functional difference is the conformational variation between the C-terminal domains of the two toxins, particularly in the highly conserved undecapeptide termed tryptophan rich motif. Here, we present the crystal structure of suilysin, a CDC toxin from the infectious swine pathogen Streptococcus suis. Like PFO, suilysin does not require a host receptor for hemolytic activity; yet the crystal structure of suilysin exhibits a similar conformation in the tryptophan rich motif to ILY. This observation suggests that the current view of the structure-function relationship between CDC proteins and membrane association is far from complete.
Amino Acid Sequence ; Animals ; Bacterial Toxins ; chemistry ; Bacteriocins ; chemistry ; Cholesterol ; chemistry ; Crystallography, X-Ray ; Cytotoxins ; chemistry ; Hemolysin Proteins ; chemistry ; genetics ; Molecular Sequence Data ; Point Mutation ; Protein Structure, Tertiary ; Sequence Alignment ; Streptococcus suis ; metabolism ; Swine

Objective:To summarize the construction and practice of the compulsory postgraduate course "reproductive genetics in gynecology and obstetrics" in Department of Gynecology and Obstetrics, Peking University, and to evaluate the teaching effect of this course.Methods:A total of 139 postgraduates who studied in Department of Obstetrics and Gynecology, Peking University, from 2019 to 2021 were enrolled as subjects, and a syllabus was constructed through a top-level design based on the "biological-psychological-social medicine pattern", with the teaching objectives of reproductive genetics theory, clinical translation, genetic counseling methods, and research advances. The teaching effect was evaluated by analyzing teaching assessment results, teaching evaluation feedback, and teaching achievements. SPSS 26.0 software was used to perform the t-test and the chi-square test. Results:The written test score, usual performance score, and total score of the postgraduate students in 2021 were higher than those in 2019 and 2020 [(73.50±8.19) vs. (70.94±14.90); (68.60±2.82) vs. (68.22±4.58); (90.58±4.18) vs. (89.49±7.60)], with significant differences in written test score and total score ( P<0.05). There was a high degree of satisfaction with the feedback of teaching, and 85.61% (119/139) of the students selected "great satisfaction"; in particular, there were increases in the degree of satisfaction with expanding research ideas and reflecting the advances in this discipline, but with no significant difference. There was a significant increase in the number of published articles. Conclusion:For the active implementation of the course of "Reproductive Genetics in Obstetrics and Gynecology", improving the teaching process in a planned and step-by-step way through a top-level design in advance can help to expand research ideas for future research work among postgraduates in obstetrics and gynecology and promote the sustainable development and improvement of the teaching of the emerging interdisciplinary discipline of reproductive genetics in obstetrics and gynecology.