Laboratory animal science, as a supporting discipline in life science research, has been set up courses in many universities, especially in medical schools.However, due to the curriculum design was too rigidly attached to tradition, emphasis on theory and cannot adapt to the development of natural science.In this paper, aiming to enhance the teaching effect, we focus on the practice, advance with the times, combine with customized teaching reform of laboratory animal science curriculum, teaching content and teaching forms.

Objective To investigate the significance of enzyme-linked immunosorbent assay (ELISA) in cross-match test of donor-recipient before organ transplantation.Methods HLA glycoproteins were prepared by solubilizing the lymphocytes of donor with a non-ionic detergent, and bond to the monoclonal antibody specific for HLA class Ⅰ or Ⅱ immobilized in the ELISA tray. ELISA with addition of recipients' serum and complement-dependent cytotoxicity (CDC) were compared.Results Cross-match test was performed by two methods for lymphocytes of 40 donors and sera of 72 recipients. All samples were successfully tested. The results of one pair of donor-recipient cross-match test by two methods were different.Conclusion ELISA for cross-match test is simple, convenient and time-saving, but more sensitive and specific than CDC.

Objective To study the distribution and tumor targeting property of anti-human prostate specific membrane antigen(PSMA) monoclonal antibody Ed-5 labeled with ~ 131 I in nude mice bearing human prostate cancer. Methods The nude mice xenografted models of human prostate cancer were established. Anti-human PSMA monoclonal antibody Ed-5 was labeled with ~ 131 I. ~ 131 I-Ed-5 was injected through the tail vein of nude mice bearing human prostate cancer. r-ray scintigraphy was performed at 24,48,96 and 144 hours after injection. And then the mouse were killed, the radioactivity ratio of tumor and non-tumor (T/NT), percentage of injected dosage of every gram tissue (%ID/g) were measured, and the bio-distribution of the labeled antibody was determined. Results The tumor imaging was clearly visualized at 96 hours after ~ 131 I-Ed-5 injection, and the tumor brim was clearer along with time prolonging. Radioactivity assembled in the tumors 24 to 144 hours after ~ 131 I-Ed-5 injection. The tumor's %ID/g at 96 hours after injection was 32.30, the radioactivity ratio of tumor and various tissues reached a maximum, and that of tumor/muscle was 7.08. Conclusion Ed-5 exhibited excellent immunoreactivity and tumor targeting property, and had application potential in targeting diagnosis and therapy of prostate cancer.

Objective To evaluate the practical value of Serology, PCR-SSP and PCR-SSOP for HLA-B typing. Methods A total of 30 samples, the blood of patients and donors waiting kidney transplantation, were used in the study. HLA-B typing was performed by one-step monoclonal antibody typing, micro PCR-SSP typing and PCR-SSOP reverse hybridisation. Results All samples were successfully typed by three methods. The difference between serological and PCR-SSP typing was 13%. The difference between PCR-SSOP and PCR-SSP typing was 3%. Conclusion Serology is high discrepancy rate and low-resolution, but cheap, simple and rapid. PCR-SSP and PCR-SSOP typing are high specific and accuracy. SSP is suitable for several samples, and SSOP is for lots of samples simultaneously although it needed long time.

Objective:Preformed anti-human leukocyte antigen(HLA)specific antibodies are a major risk for antibody-mediated rejection.The aim of this study is to detect and analyze anti-HLA specific antibodies in sera of renal transplant candidates for evaluating the status of presensitization.Methods:A total of 3 500 patients awaiting renal transplantation in our hospital from 1998 to 2007 were included in the study.Panel reactive antibody(PRA)in sera of 694 candidates was detected by complement-dependent cytotoxicity(CDC)before March 2000,then the sera of other 2 806 recipients were screened by enzyme-linked immune absorbent assay(ELISA)alternatively.The polymorphism,specificity and relevant factors of anti-HLA antibodies were analyzed.Results:Only IgG type of antibodies specific to HLA classⅠcould be detected by CDC,whereas both anti-HLA classⅠand classⅡ of IgG class could be found out by ELISA.Lower PRA positive rate by CDC(8%)was shown when compared to that by ELISA(17%).Furthermore,multiple types of specific antibodies against HLA-A(20),B(37),CW(8),DR(14)and DQ(7)were determined by ELISA.Some types of the antibodies presented higher frequencies,such as anti-HLA-A2,24,68,23 and 32;B27,56,57 and 7;DR7,4,9,13,17 and 12;CW1,2,6,4 and 8.These high frequenies of anti-HLA antibodies were somewhat different from the distribution of HLA antigens in South China population.There were significantly different positive rates of anti-HLA antibodies between the male and the female,as the male were sensitized mainly through blood infusion and the female were sensitized by either blood infusion or pregnancy.Conclusion:Specific antibodies against HLA can be detected out by ELISA accurately,whereby to find high freguencies of the antibodies and to avoid donor-recipient mismatching at HLA-loci.Detection of preformed anti-HLA and reducion of HLA-mismatched blood infusion to reduce production of anti-HLA antibodies may be the valuable pathway to improve graft survival.

Objective To investigate the effect of treatment for frozen shoulder by manipulation release combined with hydraulic pressure distension.Methods In this randomized trial,78 patients with frozen shoulder syndrome randomly divided into three groups were enrolled in the study.28 patients were treated by manipulation release under interscalene brachial plexus anesthesia.23 patients were treated by sodium hyaluronate injection into glenohumeral joint combined with manipulation release.27 patients were treated by hydraulic pressure distension combined with manipulation release.All patients were followed for six months.Effective rate evaluation combined with pain evaluation and functional evaluation were adopted for final evaluation.Results There were significant difference between pre-treatment and post-treatment in each group.Effective rate of hydraulic pressure distension combined with manipulation release was 92.6%,which was superior to manipulation release (78.6% ) and sodium hyahronate injection combined with manipulation release ( 82.6% ).There were no significant difference in functional evaluation of the three groups.Conclusion Hydraulic pressure distension combined with manipulation release under interscalene brachial plexus anesthesia is a successful treatment for idiopathic frozen shoulder.

Objective To verify the feasibility of Cas9 microinjection in frozen-thawed pronuclear embryos, based on the model of pronuclear embryos of C57BL/6J mice by in vitro fertilization.Methods After fertilized mouse pronuclear embryos cultured in vitro, one-cell and 2-cell embryos were frozen using EFS20/40 cryopreservation tube.The next day recovered and then cultured.The recovery rate and survival rate of the one-cell and 2-cell embryos were compared.The frozen-thawed and fresh pronuclear embryos were injected with Cas9 mixture and injection buffer into the cytoplasm, and then cultivated to 2-cell embryos,and the survival rate and development rate of the 2-cell embryos were compared.Results The recovery rate of frozen-thawed one-cell embryos was 92.5%, the survival rate was 92.8%, the recovery rate of 2-cell embryos was 90.5% and the survival rate was 95.8%, showing no significant difference.Furthermore, the survival rate of fresh one-cell embryos after Cas9 injection was 92.7%, the survival rate of one-cell embryos of the blank group was 97.5%.While the survival rate of Cas9 injected frozen-thawed one-cell embryos was 82.6%, and that of the blank group was 92%,showing a significant difference between the frozen-thawed injected group and other groups(P < 0.05).The development rate of 2-cell embryos after Cas9 injection was not significantly different.Conclusions Frozen-thawed pronuclear embryos can be used for Cas9 microinjection.

AIM: To analyze the difference of endonuclease domain containing 1 (ENDOD1) expression be-tween benign prostatic hyperplasia ( BPH ) tissues and prostate cancer ( PCa ) tissues and to investigate the effect of ENDOD1 on the biological function of human prostate cancer cells .METHODS: The BPH samples ( n=20 ) and PCa samples (n=21) were processed and analyzed according to the instruction of immunohistochemical (IHC) staining.The mRNA and protein levels of ENDOD 1 in the normal prostate epithelial cells and prostate cancer cells were evaluated by RT -qPCR and Western blot , respectively .The recombinant plasmids pCMV-N-Flag-ENDOD1 was constructed and was trans-fected into the human prostate cancer cells .The proliferation , apoptosis , migration and invasion abilities of the prostate cancer cells were evaluated by MTT assay , flow cytometry, Transwell migration and Matrigel invasion assays , respectively. RESULTS:The analysis of variance of the immunoreactivity score showed that PCa tissues with high Gleason score dis -played significantly lower ENDOD1expression than that with low Gleason score and BPH (P<0.05).The expression of ENDOD1 at mRNA and protein levels in PC3 cells and DU145 cells was significantly lower than that in the LNCap cells (P<0.05).The proliferation of DU145 transfected with ENDOD1 was inhibited.The flow cytometry indicated that ENDOD1 over-expression in the DU145 cells resulted in a notable increase in G0/G1 phase arrest (P<0.05), but the ap-optotic rates showed no statistical difference .The results of Transwell assay showed that migration and invasion abilities of the cells were also inhibited after transfection with over-expressing ENDOD1 plasmid (P<0.05).CONCLUSION: The expression of ENDOD1 significantly decreased in prostate cancer with high Gleaon score .Meanwhile, the ENDOD1 is spe-cifically down-regulated in androgen independent prostate cancer (AIPC) cell lines.Over-expression of ENDOD1 remark-ably inhibits the proliferation , migration and invasion abilities of AIPC .

ObjectiveTo investigate prophylaxis and treatment of bile leakage from hepatic duct anomalies after liver transplantation.MethodsWe retrospectively analyzed 3 patients with bile leakage from hepatic duct anomalies after liver transplantation in our institute.The graft procurements were combined liver-kidney harvesting.The reconstruction of the bile ducts was end-to-end anastomoses.ResultsIn the first patient with a right accessory duct joining the cystic duct,leakage of bile came from the stump of the cystic duct after anastomosis of the bile ducts.The original anastomosis was taken down,and reanastomosis was performed after plasty of the bile ducts.The patient recovered uneventfully.In the second patient with a Luschka bile duct,the biliary fistula closed spontaneously after percutaneous drainage.However,re-transplantation was performed for severe infection 7 month after the primary transplantation.In the third patient with an accessory hepatic duct from the right posterior sector joining the common bile duct,the bile duct stump which we missed leaked bile.Re-transplantation was performed because of severe complications.Conclusion Understanding the anatomy of intra- and extra-hepatic bile ducts and their common anomalies identifying the structures in the porta hepatis during preparation of the liver grafts,and looking for possible accessory hepatic ducts and aberrant bile ducts are important steps to prevent bile leakage in liver transplantation.

Objective To explore surgical opportunity and management of high-energy Pilon fractures.Methods Twenty-five cases of the high-energy Pilon fractures were treated with anatomic type plate one-staged or ses of type III fractures.8 cases were open fractures and 17 cases were close fractures.8 cases open fractures and 6 cases close fractures were treated with anatomic type plate one-staged.11 cases close fractures with serious soft tissue swell were treated two-staged delayed.The combined fibular fractures were fixed accordingly.All cases were followed up for 32 months after operation in average( range 17 to 76 months).Results According to Mazur's criteria,the surgical result was evaluated as excellent in 10 patients,goed in 11 ,fair in 3 and poor in 1.Postoperative complication ineluded infection in 2 cases,necrosis of skin in 1 case and traumatic arthritis of the ankle joint in 2 cases.Conclusion In order to get a satisfactory result,it is important to choose the right timing of surgery and correct procedure in high-energy Pilon fracture.