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Objective To explore the early diagnosis of Alport′s syndrome(AS).Methods Renal and skin biopsy was carried out in 5 patients who manifested with isolated hematuria and nephritic syndrome(NS).By using indirect immunofluorescence method,the expression of type Ⅳ collagen ? chains was detected on epidermal basement membrane(EBM) and glomerular basement membrane(GBM).Results ?_1 chains on EBM and GBM were expression in all patients,but ?_5 chains on EBM and ?_3,?_5 chains on GBM form 2 female patients were segmental expression.Thus the goal for early diagnosis was achieved.Conclusions ? chains for EBM type Ⅳ collagen and GBM type Ⅳ collegan should be investigated if condition permits for those patients with isolated hematuria,NS(steroid-resistant) and thinned GBM in electron microscopy.It can be useful for diagnosis and differenial diagnosis of AS.
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Objective To use the antisense of Platelet-derived growth factor ? chain (PDGF-?) gene to inhibit the proliferation of rabbit vascular smooth muscle cells(VSMCs) in order to provide in situ basis on prevention and treatment of human restenosis. Methods A rabbit restenotic vascular model was constrcted and an antisense designed according to PDGF-? cDNA sequence as a drug was used to observe its effects on the expression of proliferating cell nuclear antigen and in situ expression of platele-derived growth factor ? chain by the VSMCs and the formation of neointima after injury. Results The antisense could significantly inhibit the expression of proliferating cell nuclear antigen and downregulate in situ expression of PDGF-? mRNA by intimal VSMCs one week after injury with inhibitory rates of 93.44% and 88.40%, respectively. Conclusion The antisense designed could inhibit the formation of neointima after injury through downregulating the expression of PDGF-? mRNA by local VSMCs, which provides the experimental basis of the antisense for the prevention and treatment the human restenosis.
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Objective: To clone human T cell receptor (TCR) ? chain gene and express its encoding protein by baculoviral expression system in insect cells. Methods: TCR ? chain cDNA was cloned from normal human peripheral blood mononuclear cells (PBMC) by RT-PCR and inserted into baculoviral transfer vector. This vector was co-transfected with baculovirus into insect sf 9 cells. The recombinant protein expressed was identified by SDS-PAGE and flow cytometry with mouse anti-human ? chain monoclonal antibody. Results: Human TCR ? chain cDNA cloned and inserted into baculoviral vector specifically expressed protein in the insect sf 9 cells, accounting for about 11% of the total protein yield in the supernatant of cell lysate. The molecular weight of the recombinant protein determined by SDS-PAGE was identical to what we anticipated. The insect cells transfected with recombinant baculovirus were demonstrated by intracellular labeling flow cytometry to express ? chain protein. Conclusion: Human T cell receptor ? chain gene was successfully cloned from human PBMC, and its encoding protein was highly expressed in insect cells. The TCR ? chain protein obtained by bioengineering technique is useful for in depth biological function study.
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AIM: To detect the association between the polymorphism of Fc receptor ? chain gene at position-29 in promoter and systemic lupus erythematosus(SLE). METHODS: The genotypes at position -29 in promoter of Fc receptor ? chain gene were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method in 180 patients with SLE and 140 ethnically matched controls in southern China. RESULTS: The frequencies of TT genotype(33.3%) and T allele (54 4%) at position -29 in patients with SLE were significantly higher than those in controls (17 9%, respectively), whereas, the frequencies of GG genotype (24 4%) and G allele (45 6%) in patients with SLE were remarkably lower than those in controls (31 4% and 57 1%, respectively) ( P 0 05) . CONCLUSION: Our results indicate that the T allele at position -29 in promoter of Fc receptor gene probably contributes to the susceptibility to SLE, but does not play a role in the occurrence of lupus nephritis.
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AIM:To establish a real-time PCR technique for detection and quantification of TCR ? chain expression and to investigate TCR ? chain expression level in patients with chronic myeloid leukemia(CML).METHODS:Real-time PCR with SYBR GreenⅠ technology was used for detecting TCR ? chain expression level in peripheral blood mononuclear cells from 30 patients with CML and 30 normal individuals.?2-microglobulin gene(?2M) was used as an endogenous reference.Relative changes in TCR ? chain expression level were used by the 2-Ct method between patients with CML and normal individuals.RESULTS:The SYBR GreenⅠ real-time technique for quantitative detection of TCR ? chain expression levels was established successfully.The expression level of TCR ? chain in 18 patients with CML was reduced.However,the TCR ? chain expressed increased in 12 patients with CML.CONCLUSION:The TCR ? chain expression level is divided into down expression(60%) and over expression(40%) groups,and the down expression of TCR ? chain might related to cellular immunodeficiency in most of CML patients.
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The distributioa of Se in Hb and the effects of Se intake and its chemical forms oil the distribution were studied. The results showed that Se in human Hb was all located in globin and divided equally in a and ? chains. However, a part of Se could fall from Hb during the process of isolation when more Se as SeMet was taken and "excess" Se appeared in Hb. It was implicated that there were at least two kinds of linkage between Se and Hb --tight and loose.Regarding the components of amino acid in ? and ? chains, we assumed that a tight linkage between Se and Hb was in the form of SeMet, which was incorporated into Hb by dietary Se during the synthesis of Hb; the loose linkage was formed by a part of "excess" Se and CyS, especially No.93 CyS in ? chain, being unstable sclcnotrisulf ide or selenopersulfide, which broke easily by reduction during isolation. They might be utilized first by human body when Se was required.