Insufficient supply of organ for allotransplantation made the study on finding new organ resources from animal progress. Pig is regarded as one of the optimal donor animals for human. The major obstacle in this field is hyperacute reaction (HAR), which is triggered after the xenogenic natural antibodies preexisting in recipient blood combine to the antigens on the surface of the endothelium and activate the complement system. alpha-Galactose residues (alpha-Gal) on the endothelial cell have been identified as the major xenoantigens. NJZ Pig has been closely breed since 1938, whose family history is clear. Tissue samples from heart, liver, kidney, pancreas, lung, small intestine, skin, spleen, thymus and lymph node were obtained and embedded in paraffin. The sections were performed the immunohistochemical staining with the sera from health volunteers (including all the blood types) as the primary antibodies as well as the biotin labeled bandeirae simplicifolia I isolectin B4 (BS I-B4), which has specific affinity to alpha-galactose. All the staining sections were compared with the tissues digested with alpha-galactosidase. There was no difference between the antigens recognized by sera of different blood types. alpha-Gal was still the major xenoantigen on the endothelial cells. There might exist non-alpha-Gal antigens on the distal convoluted tubules and collecting tubules of the kidney. There was no alpha-Gal distributing on the secreting part of pancreas, either the islet cells or the matrix cells, but surely on pancreatic duct and vessels. All the antigenity was destroyed after the enzyme digestion except that the small intestine gland still positive with the BS I-B4. alpha-Gal is the major xenogenic antigen in NJZ Pigs. There exist some unknown antigens on the distal convoluted tubules and collecting ducts of the kidney. The blood type of recipient is not the first affair to be considered in pig-to-human xenotransplantation. The specificity of BS I-B4 for the alpha-galactose needs more detail research.
OBJECTIVE: To investigate the expression and distribution of xenoantigen in intervertebral disk of Chinese banna minipig inbred line, and to study the availability of xenograft transplantation of intervertebral disk. METHODS: Samples of intervertebral disk were collected from six Banna pigs of 8 to 11-month-old. The fixation, embedment and slice were performed. α-Gal specific binding lection (BSI-B4) were used as affinity reagents and affinity-immunohistochemistry assays (SABC methods and DAB stain) were conducted to detect the expression and distribution of xenoantigen (α-Gal). RESULTS: alpha-Gal was found in chondrocyte cell and chondrocyte-like cell in intervertebral disk which have the positive yellow-stained particulate aggradation. There was no stain in the matrix, elastic fiber and collagen fiber. CONCLUSIONS: The distribution of xenoantigen is locally in the tissue of intervertebral disk and its expression is weak. This suggests that the intervertebral disk of Banna pig may be alternative donor for xenotransplantation.
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)
Prostate cancer ranks second among the causes of death of malignant tumors in middle-aged and elderly men. A considerable number of patients are not easily detected in early-stage prostate cancer. Although traditional imaging examinations are of high value in the diagnosis and staging of prostate cancer, they also have certain limitations. With the development of nuclear medicine instruments and molecular probes, molecular imaging is playing an increasingly important role in the diagnosis and treatment of prostate cancer. Positron emission tomography and computed tomography (PET/CT) using prostate-specific membrane antigen (PSMA) as a probe has gained increasing recognition. This article will review the latest progress in the application of PET/CT using probes for targeting PSMA to imaging and treatment of prostate cancer, in order to provide a theoretical basis for the application of probes for targeting PSMA in the diagnosis and treatment of prostate cancer.
Objective To analyze inducing factors and clinical characteristics of deep venous thrombosis (DVT) and to explore clinical value of soluble cell surface differentiation antigen 40 ligand (sCD40L) in early diagnosis of DVT. Methods The patients with the DVT of lower extremity who had not received the anticoagulant and thrombolytic therapy in the Nanchong Central Hospital from January 2012 to January 2017 were collected, these patients were divided into an early-acute stage, mid-acute stage, late-acute stage, and subacute stage according to the clinical course of DVT. The sCD40L expression in the peripheral blood of DVT patients were detected by the enzyme linked immunosorbent assay. Results There were 100 patients with the DVT were included, including 31 cases of early-acute stage, 26 cases of mid-acute stage, 21 cases of late-acute stage, and 22 cases of subacute stage; 66 patients with the peripheral type, 28 patients with the central type, and 6 patients with the mixed type. ① The fracture, malignant tumor, long time in the bed following the thoracic or abdominal operation, joint replacement, and caesarean section were the successively main risk factors of the DVT. ② The early-acute stage of DVT was more common in the fracture patients, the mid- and late-acute stage of DVT often occurred in the joint replacement sufferer, and the subacute stage of DVT was usually found in the malignant tumor patients. ③ The sCD40L expression in the patients with the different stage DVT was signifiantly higher than that in the control group (20 healthy people in the physical examination, P<0.05). Furthermore, there was a significant difference in the different stage DVT patients (F=26.57, P=0.02), that is, the expression of sCD40L was the highest in the early-acute stage of DVT, and then gradually reduced (P<0.05). ④ The sCD40L expression had a significant difference among the central type DVT, mixed type DVT, and peripheral type DVT (F=12.51, P=0.02), which in the peripheral type DVT was significantly higher than that of the central type DVT (P<0.05) and mixed type DVT (P<0.05), but had no difference between the central type DVT and the mixed type DVT (P>0.05). ConclusionsCD40L might act as a blood index of early diagnosis and judgement of extent of DVT, especially be helpful in early-acute stage of DVT.
Objective To study the immunological tolerance induced by blocking the second signal of T cell with extrinsic cytotoxic T lymphocyteassociated antigen 4 immuno globlin(CTLA4-Ig). Methods Fifty-four BALB/C mice, inbred strains, were employed as recipients of bone allografts, using a model of heterotopic muscle pouch. The 54 mice were divided into 3 groups and18 for each group. The first group, in which the donor was C57BL/6 with intraperitoneal injection ofL6(as a control), was named AL group. The second group,also C57BL/6 with injection CTLA4-Ig, was named AC group. The third group,homologous BALB/C with injection of PBS buffer solution, was named AB group.The serum antibody, lymphocyte proliferation of the second stimulation by splenic cell and bone supernatant of donor, the analysis of lymphocyte subsets, a regraft experiment and histology were determined 2, 4 and 6 weeks after transplantation. The second transplantation was to regraft C57BL/6(BC group) and C3H(BHgroup) mice respectively after first 12 mice being transplantated with C57BL/6 and injected with CTLA4-Ig as to detect donor-specificity of immunological tolerance. Results Compared with AB group, AL group created more intensive immune rejection: CD4 T cell subsets(Plt;0.05), the serum antibody(Plt;0.05) and lymphocyteproliferation of the second stimulation by splenic cell and bone supernatant ofdonor (Plt;0.01 and 0.05) were significantly increased. However, the results of AC group showed that CTLA4-Ig significantly inhibited the immune rejection: CD4T cell subsets(Pgt;0.05), the serum antibody (Pgt;0.05), and lymphocyte proliferation of the second stimulation(Pgt;0.05) were similar to those of AB group. Histological observation of AC group showed that lymphocyte infiltration disappeared,cartilage and new bone formed, and bone marrow cavities emerged. A regraft experiment showed that CD4 T cell subsets (Plt;0.05) and lymphocyte proliferation of the second stimulation by splenic cell and bone supernatant of donor(Plt;0.05), BC group was significantly lower than those of BH group. So theimmunological tolerance induced by CTLA4-Ig was of donor-specificity. Conclusion The immunological tolerance induced by CTLA4-Ig was prolonged for 6 weeks. This study provides a brand-new path for bone transplantation, which can be helpful to other organ transplantation.
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 probe the association between HLA-B27 subtype and acute anterior uveitis(AAU). Methods The HLA-B27 gene of 49 AAu patients were inspected by means of polymerase chain reaction(PCR), and the B27 subtypes of AAU patients were analyzed by means of DNA sequencing and simulated nucleotide hybridization in computer. Results Twenty-nine(59.39%)of 49 AAU patients were B27 positive, and among them 13 (44.00%)were B2704 and 16(56.00%)B27052 in subtypes.No statistically significant difference in most clinical features were found between the patients with B2704 and B27052.In B27 pasitive patients,8 (seven is B27052 and one is B2704) of them were complicatde with ankylosing spondylitis(AS). Conclusions The AAU patients with either B2704 or B27052 have similar clinical feature.The B27052 gene may be an important factor for complicated AS in AAU patients. (Chin J Ocul Fundus Dis, 1999, 15: 139-142)
Objective To study degradation of the antigen-extracted meniscus in PBS solution with no enzyme or with different enzymes. Methods Four types of enzymes (collagenase, hyaluronidase, trypsin, papain) were used to enzymolyze the antigen-extracted meniscus and the fresh meniscus for 3, 7, 15 and 30 days (37℃). The antigenextracted meniscus and the fresh meniscus were immersed in PBS solution (37℃) for 30 days. Weight loss measurement, UV spectrophotometry, and scanning electron microscopy (SEM) were used to characterize the degraded materials. Results The two types of the materials were remarkably digested under the enzymes, especially under trypsin. The degradation curves showed that the antigen-extracted meniscus was enzymolyzed less than the fresh meniscus. The degradation products were grouped as amino, peptide, and polyose by the analysis. Both of the materials could hardly behydrolyzed in PBS solution without the enzymes. The four different enzymes had different surface morphologies under the examination of SEM. Conclusion The antigen-extracted meniscus is enzymolyzed more slowly than the fresh meniscus in vitro, and the result can be used as a guideline to the further research.
Objective To summarize the advancement on Lewis-Selectin metastasis pathway and anti-tumor treatment of cimetidine in colorectal cancer.Methods Domestic and international publications involving Lewis-Selectin metastasis pathway and antitumor role of cimetidine in recent years were collected and reviewed. Results The expression of sialyl Lewis X (sLeX) was most significant in Lewis-Selectin metastasis pathway in colorectal cancer. Cancer cells could interact with E-selectin and P-selectin of activated vascular endothelial cells by sLeX ligand on the cell surface, thereby prompting invasion and metastasis. Cimetidine could inhibit the growth of tumor, improve immune response of the host to tumor cell, inhibit angiogenesis of tumor and metastasis. The curative effect of cimetidine on colorectal cancer patients with positive sLeX expression was better than that with negative sLeX expression. Conclusion Lewis-Selectin metastasis pathway is one of the important metastasis pathway in colorectal cancer. Cimetidine can prevent metastasis of colorectal cancer by blocking sLeX expression.