Objective To investigate the expression of T cell receptor (TCR) Vβ8.3 gene on CD4+ T lymphocytes in the rats with experimental autoimmune uveoretinitis (EAU). Methods Eighteen Lewis rats were divided into EAU, complete Freund′s adjuvant, and the control group. Inter photoreceptor retinoid-binding protein (IRBP) R16 peptide was synthesized using Fmoc procedure for induction of EAU. Magnetic absorption cell sorting (MACS) me thod was used to isolate the CD4+T lymphocytes from the spleen of the rats. Flow cytometry was used to monitor the efficiency of isolation. The expression of TCR Vβ8.3 gene segment on CD4+T lymphocytes was determined by fluorescent quantitative polymerase chain reaction. Results EAU was successfully induced in the Lewis rats immunized with IRBP R16 peptide. The proportion of CD4+T lymphocytes isolated by means of MACS was statistically higher than that before isolation (P＜0.001). The expression of TCR Vβ8.3 gene segment on CD4+ T lymphocytes in EAU rats was significantly higher than that in the control (P＜0.05). Conclusions There is a predominant usage of antigen-specific TCR Vβ 8.3 gene in EAU rats induced by IR BP R16 peptide, which may serve as a target for immunotherapy of EAU. (Chin J Ocul Fundus Dis，2004，20：165-167)
Purpose To investigate whether experimental autoimmune uveitis can be induced equally in different rats by urea soluble fraction of bovine melanin-associated antigen(USF-BMAA),and,if so,difference among them. Methods Lewis rats,F344 rats,Wistar rats were immunized with USF-BMAA emulsified with complete Freud is adijuvant and Bordelella pertussis to induce experimental autoimmune uveitis.The animal models were investigated clinically and histopathologically and compared with each other. Rusults Experimental autoimmune uveitis could be induced in Lewis rats,F344 rats and Wistar rats with US-BMAA.Clinical and histopathalogical examination showed that bilateral ocular inflammation developed after immunization 9-13 days.Although inflammation was mainly located in anterior uvea,a mild focal choroiditis was noted in those with severe anterior inflammation.No inflammation was observed in the retina and pineal gland.Experimental autoimmune uveieis induced with USF-BMAA was similar to experimental autoimmune anterior uveitis incited with BMAA presented by other authors.Inflammation induced with USF-BMAA in F344 rats and in Lewis rats was quite similar in the severity and course of the model.But the inflammation was less in Wistar rats compared with that in Lewis rats and F344 rats. Conclusion Experimental autoimmune anterior uveitis was successfully induced with USF-BMAA in Lewis rats,F344 rats and Wistar rats.The difference with regard to the severity among these aminals were propably attributed to their genetic bankground. (Chin J Ocul Fundus Dis,1998,14:149-152)
【Abstract】Objective This study was conducted to build experimental model of orthotopic liver transplantation in rat (ROLT) with the character of acute rejection; and to study the effect of cytotoxic T lymphocyte antigen 4 immunoglobulin G (CTLA4Ig) on prevention of rejection and the induction of immune tolerance of ROLT. Methods Build model of Wistar→SD ROLT(performed by the two cuff method) with character of acute rejection.Recipients were injected with CTLA4Ig 75 μg per ROLT into abdominal cavity after 2 days of operation. Contrast was made with no treatment group, the clinical characters, the liver function, the transplantated liver pathologic character and the concentrations of TNFα in serum were observed and measured on postoperative day 7. In treatment group, all above observation were done on postoperative month 4. Above all, determination of the effect of CTLA4Ig on preventing acute rejection and inducing tolerance in ROLT was observed.Results ①Recipients (no treatment group) died one by one within 6th～14th days; pathologic character of rejection in transplantation liver could be found; ② In treatment group, on postoperative day 7 and month 4, no clinical rejection character and no pathologic character of rejection in transplantation liver were found and serum concentration of cytokins related to TNFα found lower than that of contrast group(P＜0.05), and serum concentration of ALT、AST、TBIL、DBIL found lower too(P＜0.05); But serum concentration of TP and Alb was found higher than that of contrast group(P＜0.05). Conclusion ① CTLA4Ig treatment alone inhibits the rejection responce in ROLT. ② CTLA4Ig treatment can Prevent rejection and induce immune tolerance in ROLT model with characters of acute rejection; the serum concentration of cytokins related to TNFα can probably be used as evaluation standard of rejection in ROLT rejection.
Objective To establish a eukaryotic cell line that can express soluble human leucocyte antigen G1(sHLA-G1) stably. Methods The recombinant plasmid pcDNA3-sHLA-G1 is transfected by a novel nonviral, electroporation-based gene transfer method termed nucleofection into the host cell lymphoblastoid cell line (LCL)721.221 which does not express any HLA-classical I molecules. After selection by G418, the cell line stably expressingsHLA-G1 is identified by RTPCR and Dot-ELISA with HLA-G1 specific monoclonal antibody MEM-G/9. Results The efficiency of transfection for LCL721.221 is about 14% by nucleofection. The specific band forsHLA-G1 was found by RT-PCR assay from the transfections and the protein ofsHLA-G1 in the supernatant of the transfections was detected by Dot-ELISA assay. Both confirmed that the eukaryotic cell line expressingsHLA-G1 has been established successfully at genic and proteinic levels. Conclusion In this study, the eukaryotic cell line expressingsHLA-G1 have been established successfully by nucleofection.
Objective To investigate the relation of Human Leukocyte Antigen-DRs to Pre-eclampsia/eclampsia （PE/E） by reviewing the observational studies on PE/E. Methods We searched the MEDLINE/PubMed, EMBASE, The Cochrane Library and CBMdisc to July 2005, by combining free text with MeSH words. We assessed the quality of included studies, extracted and analyzed data. Results The odds ratio of fetal-maternal HLA-DR4 antigen frequency in case group versus control group was 2. 60 （95% CI 1.87 to 3.60） with statistical significance. The antigen frequencies of＇other fetal-maternal HLA-DRs in case and control groups were not statistically significant. The antigen frequencies of the couple HLA DRs were not statistically significant between case and control groups. We found that neither the HLA-DR sharing between the couples nor between fetus and mothers in case and control groups were statistically significant. Conclusions The antigen frequencies of HLA-DRs between the couples may have no association with the development of PE/E. The fetal gene types may be related to the development of PE/E. The HLA-DR sharing in mothers and fetus and the couples may have no association with the development of PE/E.
Serum tumor markers CEA, CA19-9, CA72-4 and Helicobacter pylori (H.pylori) antibodies were measured in 162 patients with gastric cancer. CEA, CA19-9 and CA72-4 had sensitivities of 24.0%, 35.5% and 21.9% respectively. CA72-4 provided 100% specifity, compared to 77% and 93% for CA19-9 and CEA. The positive predictive value (PV) in CEA, CA19-9 and CA72-4 was higher than negative PV. Serum CA19-9 and CA72-4 levels rose in tumor of ＞5.0cm in diameter. The CA19-9 increased remarkably when the deeper stomach wall was invased. The significantly elevated CEA, CA72-4 and CA19-9 levels were found in patients who had nodal involvement in more than 50% and distant metastasis. However, the increase of CEA, CA19-9 and CA72-4 were found in undifferentiated tumor. Antibodies to H.pylori were detected in 54% of patients but in only 22% control subjects. A significant association was found between H.pylori infection and gastric cancer (odds ratio=3.75; 95% confidence interval=2.11-5.41, Ｐ＜0.01). Conclusions: CEA, CA19-9 and CA72-4 have higher specifity but lower sensitivity in diagnosis of the gastric cancer. The levels of CEA, CA19-9 and CA72-4 are significantly associated with the diameter, the depth of invasion, nodal involvement, distant metastasis and cell differention. Infection with H.pylori may be an important cause of gastric cancer.
【Abstract】Objective To compare the reliability of serum tumour specific growth factor (TSGF) with carcinoembryonic antigen (CEA) in the diagnosis of tumour. Methods The patients were divided into two groups according to malignancy and benignity. In benignity, the patients were subdivided into inflammatory and non-inflammatory groups. The levels of TSGF and CEA in the two groups were measured. Results The positive rate of TSGF and CEA in malignant group was 67.41% and 38.84% respectively; that in benign was 24.56% and 2.63% respectively, in which the inflammatory group was 32.35% and 5.88% respectively, and in non-inflammatory group was 18.25% and 0% respectively. The positive rate of TSGF and CEA was higher in malignant than in benign group (P＜0.005). The positive rate of TSGF was higher than CEA in malignant (P＜0.005) and inflammatory group (P＜0.005). Conclusion Serum TSGF is a useful blood marker in the diagnosis of patients with malignancy, and is a more sensitive and broad-spectrum marker than CEA for the diagnosis of tumours. CEA is more specific than TSGF for the diagnosis of tumours. Combined measurement both TSGF and CEA will enhance the diagnostic rate.
Objective To investigate the effects of chitosan on the cell cycle of the human fibroblasts and on the Ki-67 antigen expression in vitro and to investigate the mechanism of chitosan preventing the postoperative tissue adhesion. Methods The cultured fibroblasts were treated for 48 hours with 0，0.01，0.1，1.0，10.0 mg/ml of chitosan, respectively;then, the cell cycle of the fibroblasts was measured by the flow cytometry. The cultured fibroblasts were treated for 24 hours with the chitiosan at the above concentrations; then, the Ki-67 antigen in the cell nucleus was detected with the immunohistochemical staining toobserve its expression. Results The growth of the fibroblastswas obviously suppressed by chitosan, especially in the cell morphology. When the concentrations of chitosan were 1.0 mg/ml and 10.0 mg/ml, the percentages of the fibroblasts in the proliferation stage were 32.3%±5.2% and 14.7%±2.9%, respectively,which were significantly smaller than the percentage of the fibroblasts when the concentration of chitosan was 0 mg/ml (the control group) (41.9%±5.8%， P＜0.05). When the concentrations were 0.01 mg/ml and 0.1 mg/ml, the percentages of the fibroblasts in the proliferation stage were 39.0%±6.0% and 35.5%±3.4%, respectively, which were smaller than that of the control, but not significantly different from that of the control (P＞0.05). When the concentrations of chitosan were 0.1 mg/ml,1.0 mg/ml and 10.0 mg/ml, the percentages of the fibroblasts that had the positiveKi-67 antigen were 37.3%±3.4%, 30.5%±6.2% and 17.8%±3.0%，respectively, which were significantly smaller than that of the control (57.6%±8.9%， P＜0.05). When the concentration was 0.01 mg/ml, the percentage of the fibroblasts that had the positive Ki-67 antigen was 54.1%±8.0%, which was smaller than that of the control, but not significantly different from that of the control (P＞0.05). ConclusionChitosan can inhibit the proliferation of the fibroblasts and increase the percentage of the fibroblasts in the quiescent stage, which can be considered as one of the mechanisms that chitosan can prevent the postoperative tissueadhesion.
ObjectiveTo observe the expression and transcription of MART-1 in human uveal melanoma cell lines 92-1, 92-2, Ocm3, Me1285, as well as the possible effect of methylation on its expression.MethodsThe cell lines 92-1, 92-2, Ocm3 and Mel285 were cultured routinely and tested for MART-1 expression at protein and mRNA level by FACS analysis, Western blot and RT-PCR respectively. Methylation status of the MART-1 promoter region in all the cell lines were checked by Southern blots of DNA digested with methylation sensitive restriction enzymes.ResultsAs observed in FACS analysis and Western blot, 92-1, 92-2 and Ocm3 were MART-1 positive cell lines while Me1285 was negative cell line. Consistent with protein analysis, 92-1 and Ocm3 cell lines showed MART-1 specific PCR products and there was no product in Me1285 cell line in RT-PCR. The MART-1 positive cell lines, 92-1, 92-2, and Ocm3 show methylation at the MspI/HpaⅡ site, and the NruⅠ sites of all positive cell lines are not methylated. The MART-1 negative cell line Mel285 shows hypermethylation at the NruⅠsite and the MspⅠ/HpaⅡ site is not methylated.ConclusionsMART-1 could be expressed in human uveal melanoma cell lines 92-1, 92-2 and Ocm3. The change of methylation status of MART-1 promoter may correlate with the transcription of MART-1.
Objective To investigate the effect of alltrans retinoic acid (atRA) on proliferative artery disease after heart transplantation. Methods Heterotopic heart transplantation model was established by Ono model with 16 inbred healthy male Wistar rats as donors and 16 SD rats as recipients. The rats were divided into chronic rejection group and atRAtreated group by complete random design, and there were 8 rats in each group. Rats in chronic rejection group were given Cyclosporine A 10 mg/(kg·d) by subcutaneous injection after operation, and those in atRAtreated group were given Cyclosporine A 10 mg/(kg·d) in the same way and atRA 10mg/(kg·d) by gavage. The transplanted hearts of rats were taken out 60 days after the transplantation. HE stain, masson stain and Van Gieson were done to analyze the rejection of transplanted hearts, the degree of vascular stenosis and myocardial fibrosis respectively.Immunohistochemistry was used to test proliferating cell nuclear antigen (PCNA). Results The area of myocardial fibrosis in chronic rejection group was obviously larger than that in atRAtreated group（63.99%±11.91% vs.34.68%±6.34%）, and there was significant difference between two groups（t=8.377，P=0.000）. The index of vascular stenosis in chronic rejection group was higher than that in atRAtreated group（62.86±17.18 vs. 40.10±8.20）. Vascular stenosis in atRAtreated group alleviated significantly, and there was significant difference between two groups（t=3.913, P=0.006）. The PCNA positive cells in chronic rejection group were obviously more than that in atRAtreated group（60.17±17.74 vs. 33.96±8.65）, and there was significant difference between two groups（t=5.387, P≤0.001）. There was a positive correlation between the PCNA positive cell ratio and the index of vascular stenosis（r=0.854, P=0.007）. Conclusion Alltrans retinoic acid can inhibit vascular disease after heart transplantation by cell proliferative pathway.