Expression of TCR Vβ subfamily in patients with idiopathic thrombocytopenic purpura

AIM: To investigate the expression of TCR Vβ subfamily in T cells in patients with idiopathic thrombocytopenic purpura (ITP). METHODS: The TCR Vβ 24 subfamily genes were amplified in peripheral blood mononuclear cells from 5 cases with ITP using RT-PCR, to observe the usage of TCR Vβ repertoire. 10 normal individuals served as controls. RESULTS: Only 4-11 Vβ subfamily in T cells were identified in ITP cases. The frequent expressions of Vβ subfamilies were Vβ2 (100%), Vβ3 (100%), Vβ19 (80%) and Vβ21 (80%), while the expression of Vβ4, Vβ6-12, Vβ17, Vβ20 and Vβ24 were not detected. All 24 Vβ subfamilies were detected in samples from normal individuals. CONCLUSION: The restricted expression of TCR Vβ subfamilies is one of the T-cell immune features in patients with ITP, indicating that it may be related to the disorder of cellular immune function in ITP.     

Effect of mesenchymal stem cells on secreting function of T lymphocytes from patients with idiopathic thrombocytopenic purpura in vitro

AIM: To analyze the effect of mesenchymal stem cells (MSCs) on secreting cytokines by T lymphocytes from patients with idiopathic thrombocytopenic purpura (ITP) in vitro.METHODS: Human bone marrow-derived MSCs were isolated by Ficoll Hypaque and cultured for proliferating to passage cells. Allogeneic T lymphocytes of ITP were isolated from peripheral blood by Ficoll Hypaque and nylon cotton column. Then the stromal feeder layers of different numbers (2×103, 1×104, 5×104 per well) of MSCs treated with mitomycin were co-cultured with above-mentioned T lymphocytes. The supernatant were respectively collected on day 2, 4 and 6 after co-culture, then the levels of IL-2, IFN-γ, IL-4, IL-10 secreted by T lymphocytes were measured by enzyme linked immunosorbent assay (ELISA) dynamically.RESULTS: The levels of IL-2 and IFN-γ secreted by T cells from ITP were higher than those from normal control (P<0.05, respectively). Inversely, IL-4 and IL-10 were lower than those in normal control (P<0.05, respectively). After co-cultured with T lymphocytes, MSCs significantly inhibited the cytokine levels of IL-2 and IFN-γ secreted by T lymphocytes from ITP or health adults (P<0.05, respectively) in a dose dependent manner (P<0.05, respectively), and the effect was more obvious when co-cultured for 4 days or 6 days than that for 2 days (P<0.05, respectively). However, MSCs significantly promoted the releases of IL-4 and IL-10 by T lymphocytes from ITP patients (P<0.05, respectively) in a dose dependent manner (P<0.05, respectively), and the effect on IL-10 was in a time dependent way (P<0.05), while the effect on IL-4 had no obvious difference among 2 d, 4 d and 6 d(P>0.05). As for health control group, when cell numbers exceeded above 1×104, MSCs obviously promoted IL-4 and IL-10 levels secreted by T lymphocytes (P<0.05) in a dose dependent manner (P<0.05), and both of the effects were more noticeable when co-cultured for 4 d or 6 d than that for 2 d(P<0.05, respectively).CONCLUSION: MSCs regulate the balance between Th1 and Th2 reaction and partly correct ITP Th1 polarization in vitro.     

Preparation and study of monoclonal antibody to matrix metalloproteinase-2

AIM: To obtain a McAb that can inhibit the function of matrix metalloproteinase-2 (MMP-2), we expressed the fibronectin-like domain of MMP-2 (MFD) in vitro and prepared a McAb against MMP-2. METHODS: The purified MFD protein was used to immunize BALB/C mouse three times. Then the spleen of mouse was taken out and hybridized with hybridoma cells SP2/0. The positive cell clones were screened with ELISA method. The subtype and tissue specificity of the McAb were identified and its effect on endothelial cell migration and tube-formation was analyzed. RESULTS: After the spleen cells of the mouse and hybridoma cells SP2/0 were hybridized, a piece of cells that continuously secreted McAb against MMP-2 was obtained and named SZ-117. The titers of this McAb in culture supernatants and ascites were 2×10^-3 and 2×10^-5, respectively. The heavy chain of the McAb belongs to IgG1 subclass. The McAb identified native MMP-2. MMP-2 existed in the stromal tissue of stomach, cholecystis, spleen, ovarian, prostate, salping and lymph node. It inhibited the invasion behavior of endothelial cells Eahy926 and pancreatic carcinoma cells 1990 and inhibited the tube-formation of Eahy926 cells. CONCLUSION: A useful tool for testing MMP-2 is obtained and it will be helpful to look for a kind of new anti-tumor material.     

High efficient expression of human endostatin in E.coli and its antiangiogensis activity

AIM: To examine the expression of human endostatin in E.coli, produce its fusion protein antibody and observe its biological activity. METHODS: Endostatin gene was amplified by polymerase chain reaction,recombined with plasmid vector pGEX-2T and induced expression with IPTG.The protein activity was tested by endothelial cell proliferation inhibitory assay.Inclusion body crudely purified was used to generate polyclonal antibody to detect its expression at mouse's liver and kidney etc. RESULTS: The protein expressed was 20kD after digestion by thrombin,it appeared the anti-angiogenesis activity and Western blotting indicated the expression of endostatin in liver and kidney of mouse. CONCLUSION: The successful expression of human endostatin and the preparation of polycolonal antibody indicated its potential application in anti-angiogenesis therapy and diagnosis tumors.     

Effects of rat tail collagen and C-erbB-2 antibody on the adhesion and proliferation in hepatocellular carcinoma cells

AIM: To explore the effect of collagen and C-erbB-2 protein on the adhesion and the proliferation in hepatocellular carcinoma cells. METHODS: Hepatocellular carcinoma cell line (HepG-2) identified to positive for C-erbB-2 gene was used to study the adhesion and the growth feature by the action of rat tail collagen and C-erbB-2 antibody.RESULTS: The action of rat tail collagen to potentiated the adhesion in HepG-2 cells was significantly but no proliferation effect was observed. C-erbB-2 antibody inhibited the adhesion and proliferation of HepG-2 cells and also abolished the potentiated effect of rat tail collagen on the adhesion in HepG- 2 cells. CONCLUSION: The signaling transduction mediated by C-erbB-2 protein was correlated to the adhesion and the proliferation of HepG-2. The blockage of C-erbB-2 gene signal transduction may be a strategic target to the treatment of liver cancer in the future.     

Expression of IgG in bladder carcinoma and the effect of the antibody against human IgG on tumor cell apoptosis

AIM: To investigate the expression of immunoglobulin G (IgG) in bladder transitional carcinoma cells (BTCC) and the effect of the antibody against human IgG on tumor cell apoptosis.METHODS: The expression of IgG of BTCC was detected by immunohistochemistry, Western blotting, enzyme linked immunosorbent assay (ELISA) and flow cytometry (FCM). The expression of IgG mRNA was tested by hybridization in situ method and RT-PCR in vitro. Antiproliferation effects of the antibody of antihuman IgG on T24 and BIU-87 cell lines were measured by MTT assay. Cell apoptosis was assessed by FCM.RESULTS: The expression of IgG was 91.1％ (51/56) in clinical cases, and 45.4％ (5/11) in normal controls. Immunohistochemistry and Western blotting showed that the IgG was significantly expressed in T24 and BIU-87. FCM indicated the IgG was mainly expressed in cytoplasma. No IgG was detected by ELISA in supernatant of cell culture medium. RT-PCR and hybridization in situ demonstrated IgG mRNA was significantly expressed in two cell lines. Under the treatment of 25 mg/L of goat nonspecific IgG and the antibody of antihuman IgG, the inhibition ratio of cell growth in T24 and BIU-87 were (4.73±3.73)% vs (24.98±3.81)% and (5.36±1.53)% vs (22.70±3.72)%, respectively (P<0.05). The percentages of apoptotic cells in T24 and BIU-87 were 2.3% vs 20.7% and 1.3% vs 15.3%, respectively.CONCLUSION: IgG is significantly expressed in bladder transitional carcinoma cell. The antibody of antihuman IgG inhibits cancer cell growth and induces tumor cell apoptosis.     

Comparison of the levels of serum prolactin and the concentrations of anti-infective factors of human colostrum in normal labor and cesarean section

AIM: To compare the difference in serum prolactin levels and the concentrations of anti-infective factors of human colostrums between normal labor and cesarean section. METHODS: The serum prolactin levels were measured at 48 h, 96 h after parturition in 100 cases of normal labor and cesarean using radioimmunoassay. Furthermore, the levels of SIgA, IgG, IgM, C₃, C₄ were tested in 100 cases at the same time using the technique of immuno-turbidimetry. RESULTS: The serum prolactin levels in normal labor tested at 48 h after parturition were significantly different with that of the cesarean section group. But there was no significant difference in serum prolatin levels 96 h after parturition between the two groups. The levels of SIgA,IgG,IgM,C₃ and C₄ of the colostrum in the cesarean group were higher significantly than those of the normal group at 48 h after parturition. (P＜0.05 or P＜0.01). At 96 h after parturition, there was no significant difference in SIgA, IgG, IgM and C₃ between the two groups except C₄. CONCLUSION:At 48 h after paturition, the serum prolactin levels of normal labor group are higher than that of the cesarean section group. The concentrations of anti-infective factors (SIgA, IgG, IgM, C₃ and C₄) of human colostrums at 48 h after paturition in cesarean section group are higher than those of normal labor group.     

番茄抗烟粉虱研究进展

烟粉虱是一种由多种生物型组成的世界性农业害虫,普通栽培番茄对其没有抗性,抗源主要来自于番茄的野生近缘种。本文对国内外番茄抗烟粉虱的种质资源,番茄对烟粉虱的抗性机制,以及遗传基础等方面的研究现状进行了总结,并提出了今后的研究重点。     

枣树抗寒性研究进展

在对枣树抗寒特性及寒害类型、枣树抗寒性生理研究进展进行综述的基础上,总结了枣树抗寒性育种面临的困难和发展现状,最后对枣树抗寒性研究发展前景做了展望。     

Detection of Glu-plasminogen in the human plasma and its clinical significance

AIM:To observe the changes of plasma content of Glu-plasminogen (plasmin, Pln) in patients during blood coagulation and/or fibrinolytic system activation.METHODS:Using specific McAb to antigenic determinant in NH₂ terminal (1-65 aa) of Pln and specific McAb to antigenic determinant in heavy chain of plasminogen, the sandwiched ELISA method was established to detect Glu-plasminogen and total plasminogen in human plasma collected from 220 normal controls and 40 patients after heart surgery.RESULTS:The average total plasminogen was (231.8±62.1) mg/L and average Glu-plasminogen was (231.9±45.8) mg/L in 220 normal controls, the ratio of Glu-plasminogen to total plasminogen (G/P) was 0.91. The ratio of Glu-plasminogen to total plasminogen in 40 patients after heart surgery was significantly lower than that in normal controls (P<0.01).CONCLUSION:The sandwiched ELISA method we established to detect Glu-plasminogen and total plasminogen in human plasma has comparatively better specificity and sensitivity for clinical use.     

我国嫁接技术防治茄子土传病害的研究进展

综述了国内茄子土传病害发生情况和嫁接技术在茄子上的应用情况,并对嫁接研究工作进行了展望。     

辣椒疮痂病抗性研究进展

辣椒疮痂病是辣椒生产中最为严重的细菌性病害之一,对辣椒的产量和品质影响较大。从病原菌的特征、分类、生理小种鉴定、抗疮痂病基因及其分子标记等几个方面总结了目前国内外的研究现状。     

黄瓜对西瓜花叶病毒病抗性的研究进展

西瓜花叶病毒（Watermelon mosaic virus,WMV）是侵染黄瓜的主要病毒之一,给黄瓜生产带来了极大的损失。本文对WMV进行了简要介绍,对黄瓜WMV的抗病性鉴定方法、抗病遗传规律、抗病种质资源和分子标记的筛选与应用进行了综述,并对其存在的问题和未来的研究方向进行了分析和展望。     

瓜类蔬菜白粉病抗性诱导及抗性遗传研究进展

瓜类蔬菜作为世界各地重要的蔬菜种类,多年来深受白粉病危害和影响,抗性诱导是减轻其危害的有效技术途径.笔者简要概述了瓜类白粉病抗性诱导及抗性遗传的研究进展,主要从生物因子、化学因子和物理因子诱导途径,以及瓜类白粉病抗性基因和抗性分子标记开发与应用等方面综述了近年来的研究成果,并对今后的研究方向提出展望.     

甜瓜抗蔓枯病育种研究进展

蔓枯病是由真菌葡萄壳梭孢引起的危害甜瓜生产的主要病害,可以侵染西瓜和黄瓜等其他瓜
类作物,造成不同程度的减产。本文介绍了蔓枯病病原菌的相关研究内容、不同接种鉴定方法对抗性鉴定
结果的影响以及我国近年来审定的部分抗蔓枯病的甜瓜新品种。甜瓜蔓枯病抗性鉴定结果容易受病原菌自
身致病力和外界环境的影响,需要通过相关的分子标记来辅助抗性种质的选育。然而,现有的一些分子标
记还存在很多不足,影响了育种的进程。因此,应该明确不同病原菌之间致病力的差异,选择合适的接种
鉴定方法,不断完善种质资源遗传多样性的评价体系,明确甜瓜种质遗传多样性的分布特点。     

Influence of Nao-xue-bao on blood coagulation and platelet aggregation in rats

AIM and METHODS:To study the effect of Nao-xue-bao at three different doses on blood coagulation,platelet aggregation by observing the changes in activated partial thromboplastin time (APTT), prothrombin time (PT), antithrombin-Ⅲ(AT-III), fibrinogen(Fng), plasminogen(Plg) and platelet aggregation(PAG). RESULTS:Compared with thrombosis group, the rats treated with Naoxuebao showed that the plasma APTT,PT were longer, and the activity of AT-III was increased. The content of Fng was reduced, TT was longer, there was a negative correlation between Fng and TT. Furthermore PAG-1, PAG-5 and PAG-M were reduced. CONCLUSION: Nao-xue-bao could inhibit thrombosis in different keys of blood coagulation.     

Relationship between renal hypertension and cardiac α1-adrenoceptor autoantibodies in rats

AIM: To examine the autoantibody against α₁-adrenoceptor and its biologic activities during the development of renal hypertension. METHODS: Renal hypertension of rat was achieved by clipped renal artery, the titre of autoantibody to α₁-adrenoceptor was detected using ELISA immunoassay. Furthermore, the biological offects of these autoantibodies on cultured cardiomyocytes were also examined. RESULTS: After two weeks of clipping renal arteries, both the frequency of occurrence and the titre of autoantibodies to cardiac α₁-adrenergic receptor were significantly increased as compared with the control of pre-treatment. The increased autoantibodies lasted for several weeks and then automatically decreased gradually to the pre-clipping level at 12 weeks. The biological effects of these autoantibodies displayed an "agonistic-like" activities on the beating frequency of cultured neonatal cardiomyocytes. CONCLUSION: Autoantibodies against α₁-adrenoceptor may play a role in the elevation of peripheral vascular resistance and in the development of cardiac hypertrophy in rats with renal hypertension.     

番茄对盐胁迫的反应及其耐盐性鉴定的研究进展

随着土壤盐渍化问题的不断加重.番茄耐盐性逐渐成为关注的热点.就盐胁迫对番茄种子萌发、生长发育、果实品质等方面的影响,以及番茄耐盐性鉴定的方法和指标等进行了综述.     

秸秆还田的土壤环境效应及研究进展

秸秆还田既可避免秸秆资源堆放浪费,也可防止其大量焚烧而造成环境污染,同时利于培肥土壤,提高土地的可持续利用。该研究结合近年来秸秆还田的相关研究进展,阐述了秸秆还田对土壤物理、化学及生物学性质的影响,进一步揭示了秸秆还田在改善土壤环境方面的重要作用,指出了当前秸秆还田技术所存在的问题,并提出建议及展望,旨在为开展秸秆还田培肥土壤的机制研究提供参考依据。     

黄瓜白粉病抗性遗传规律及分子标记研究进展

综述了国内外黄瓜白粉病抗性遗传规律、分子标记的最新研究进展。并针对研究中存在的问题进行分析,对今后的研究方向进行了展望。