Effect of Sirpα1 on the expression of nuclear factor-kappa B in hepatocellular carcinoma

BACKGROUND:Signal regulatory protein alpha1(Sirpα1) is a member of Sirps families containing four immunoreceptor tyrosine-based inhibitory motifs(ITIMs) domains in the cytoplasm of and an activated substrate of receptor tyrosine kinase(RTK),that negatively regulates the RTK-dependent cell proliferating signal transduction pathway.Previously we found that Sirpα1 was closely associated with the occurrence and development of hepatocellular carcinoma(HCC)as well as liver regeneration.Since it is unclear about the regulatory mechanisms,we established the cell line transfected Sirpα1 gene and preliminarily clarified the mechanisms by which Sirpα1 negatively regulates the carcinogenesis and development of HCC. METHODS:Liver cancer Sk-Hep1 cell was respectively transfected with plasmids of pLXSN,pLXSN-Sirpα1 and pLXSN-Sirpα1Δ4Y 2 ,screened with the drug of G418(1200 μg/ml),and various transfected Sk-Hep1 cell lines were obtained.The protein expressions of P65,P50,IκBα,cyclin D1 and Fas in various Sk-Hep1 cell lines were determined by Western blotting,and P65 and P50 were localized by the immunofluorescence technique. RESULTS:Sirpα1 could significantly upregulate the protein expression of IκBα(vs.other cell lines,P<0.05) in the Sk-Hep1 cell,and downregulate the protein expressions of P65,P50 and cyclin D1(vs.other cell lines, P<0.05)in the Sk-Hep1 cell.P65 protein expression was mainly localized in the cytoplasm in the pLXSN Sk-Hep1 cell,and in the nucleus of the Sk-Hep1 cell with mutantSirpα1Δ4Y 2 ,but in nucleus of the Sk-Hep1 cell with wild Sirpα1.P50 protein expression was localized in the cytoplasm and nucleus of the pLXSN Sk-Hep1 cell,but in the nucleus of the Sk-Hep1 cell with wild Sirpα1 and mutant Sirpα1Δ4Y 2 plasmid. CONCLUSIONS:Sirpα1 might negatively regulate and control the abnormal proliferation of liver cancer cells by influencing the protein content and localization of nuclear factor-kappa B,then influence the expression of cyclins such as cyclin D1 in the signal transduction pathway.It may be one of the important mechanisms by which Sirpα1 negatively regulates the carcinogenesis and development of HCC.     

尿蛋白-1与糖尿病肾病关系的研究

目的为了探讨尿蛋白-1(UP-)与糖尿病肾病的关系。方法用酶联免疫法测定了90例非胰岛素依赖型糖尿病(NIDDM)患者的UP-1,同时测定尿N-乙酰-β-D-氨基葡萄糖苷酶(NAG)、β2-微球蛋白(β2-mG)。结果糖尿病患者UP-1均较正常对照组增高,UP-1与尿A1b、尿NAG和β2-mG呈正相关。结论在糖尿病早期阶段,肾小管会受到损害,UP-1可作为诊断早期糖尿病肾病的敏感指标。     

孟鲁斯特对哮喘模型IL-5mRNA和IL-5蛋白表达的影响

目的 探讨白三烯受体拮抗剂孟鲁斯特(MK)对哮喘小鼠IL-5mRNA、IL-5蛋白表达的影响。方法 采用原位杂交、免疫组化LSAB法,检测哮喘小鼠及孟鲁斯特治疗后哮喘小鼠骨髓细胞IL-5mRNA的表达及肺、骨髓、脾IL-5蛋白的表达情况。 结果 孟鲁斯特可显著减轻哮喘小鼠肺组织炎症细胞浸润、细支气管痉挛、粘液分泌等;亦可减少骨髓IL-5mRNA阳性细胞数(P<0.02),并使骨髓、肺、脾IL-5蛋白表达下降。 结论 白三烯受体拮抗剂孟鲁斯特不仅使肺部炎症显著减轻,也抑制骨髓细胞表达IL-5mRNA、IL-5蛋白,提示白三烯可能通过调节炎症细胞、细胞因子合成来应答过敏原反应。     

弓形虫真核表达质粒pcDNA3/GRA1的构建及序列测定

目的 :构建弓形虫致密颗粒抗原 1 (GRA1 )真核表达质粒 ,为进一步开展DNA疫苗的保护性研究打下基础。方法 :采用PCR扩增出编码GRA1目的基因 ,用EcoRⅠ /XhoⅠ分别对扩增产物和真核表达质粒pcDNA3进行双酶切 ,将GRA1定向克隆到pcDNA3EcoRⅠ /XhoⅠ位点 ;对重组质粒进行PCR ,双酶切初步鉴定后做序列测定。结果 :特异扩增出预计的GRA1片段 ,大小为 5 73bp ;扩增产物双酶切后成功连接到pcDNA3中 ,经PCR ,双酶切及序列测定结果表明重组质粒中含有GRA1读框。结论 :成功构建弓形虫GRA1真核表达质粒。     

Use of the microorganism Bacillus stearothermophilus as a model to evaluate toxicity of the lipophilic environmental pollutant endosulfan

Microorganisms are very powerful tools for the supply of information about the toxic effects of lipophilic compounds, since an impairment of cell growth usually occurs as a result of perturbations related, in most cases, with the partition of toxicants in membranes. The thermophilic eubacterium Bacillus stearothermophilus has been used as a model system to identify α- and β-endosulfan interactions with the membrane possibly related with the insecticide toxicity. Two approaches have been pursued: (a) bacterial growth is followed and the effects of endosulfan isomers determined; (b) biophysical studies with the fluorescent fluidity probe 1,6-diphenyl-1,3,5-hexatriene (DPH) were performed to assess the effects of α- and β-endosulfan on the organization of the membrane lipid bilayer. The effects on growth were quantitatively evaluated by determination of growth parameters, namely the lag phase, the specific growth rate and the cell density reached by cultures in the stationary phase. Growth inhibition by α and β-endosulfan dependent on the concentration is diminished or removed by the addition of 2.5 m Ca²⁺ to bacterial cultures. Fluorescence DPH polarization consistently showed opposite effects of Ca²⁺ and α- and β-endosulfan on the physical state of bacterial polar lipid dispersions.     

精神分裂症患者致炎性细胞因子和酪氨酸羟化酶mRNA表达水平的研究

目的探讨精神分裂症的外周神经免疫机制及其与临床症状的关系。方法检测精神分裂症患者致炎性细胞因子白介素-1β（IL-1β）、肿瘤坏死因子-α（TNF—α）以及酪氨酸羟化酶（TH）的mRNA表达水平，采用逆转录-聚合酶链反应及半定量检测技术，分别检测39例精神分裂症患者（患者组）、25例同胞（同胞组）及30名正常对照（对照组）外周血单个核细胞（PBMC）IL-1β、TNF-α及TH基因表达水平，同时应用阳性和阴性症状量表（PANSS）评定精神分裂症患者临床症状。结果患者组、同胞组及对照组IL-1β的mRNA表达水平分别为1．52±1．01、1．52±1．09和0．74±0．38；TNF—α的mRNA表达水平分别为1．18±0．99、1．01±0．87和0．70±0．29；TH的mRNA表达水平分别为0．55±0．33、0．61±0．32和0．28±0．20。患者组和同胞组的IL-1β、TNF—α、TH的mRNA表达水平均明显高于对照组（P〈0．05或P〈0．01）。患者组IL-1β（r=0．420）、TNF—α（r=0．430）的mRNA表达水平与PANSS的-般病理症状分呈正相关（P〈0．01）。同胞组与对照组合并统计，IL-1β与TNF-α的mRNA表达水平呈正相关（r=0．847，P〈0．01）；IL-1β与TH的mRNA表达水平呈正相关（r=0．666，P〈0．01）。患者组仅IL-1β与TNF—α的mRNA表达水平呈正相关（r=0．942，P〈0．01）。结论精神分裂症患者PBMC细胞TH、IL-1β和TNF—α的mRNA表达水平高于正常，且与精神分裂症的-般病理症状显著相关。     

Renoprotective effects of combination of angiotensin converting enzyme inhibitor with mycophenolate mofetil in diabetic rats

Objective and design Previously it was shown that blocking of the renin-angiotensin system (RAS) by angiotensin converting enzyme (ACE) inhibitors, or suppression of inflammatory responses by immunosuppressive drugs such as mycophenolate mofetil (MMF) could attenuate renal injury in diabetic rats. Whether RAS blockade combined with an immunosuppressive drug provides superior renoprotection against diabetic nephropathy has not been clearly delineated. Materials Diabetes was induced by injection of streptozotocin after uninephrectomy. Treatment Rats were randomly separated into five groups: control, diabetes, diabetes treated with enalapril (an ACE inhibitor, 10 mg/kg/d by gastric gavage), diabetes treated with MMF (10 mg/kg/d by gastric gavage), or diabetes treated with a combination of both agents and were followed for 8 weeks. Methods 24 h urinary albumin excretion rate (AER) was determined, renal injury was evaluated, immunohistochemistry for ED-1 macrophage marker, intercellular adhesion molecule-1 (ICAM-1) and monocyte chemoattractant protein-1 (MCP-1) were performed, and expression of transforming growth factor (TGF)-β1 protein was determined by Western blotting analysis. Results Diabetes was associated with a considerable increase in AER. Both enalapril and MMF retarded the increase in albuminuria, which was nearly completely abrogated by combination therapy. Increased glomerular volume and tubulointerstitial injury index in diabetic rats was attenuated by treatment with either enalapril or MMF and further reduced by the combination of the two. Elevated malondialdehyde levels in renal tissue were reduced by enalapril or MMF and, more effectively, by combined enalapril with MMF. Renal overexpression of ICAM-1 was not retarded by enalapril and attenuated by MMF or combined enalapril with MMF. Combination therapy was associated with a superior suppression of diabetes-induced macrophage recruitment and overexpression of MCP-1 and TGFβ1 compared to either monotherapy in renal tissue. Conclusion The combination of enalapril and MMF confers superiority over monotherapy in renoprotection, a mechanism which may be at least partly correlated with synergistic suppression of increased macrophage recruitment and overexpression of MCP-1 and TGF-β1 in renal tissue in diabetic rats. Received 26 July 2005; returned for revision 5 October 2005; returned for final revision 9 January 2006; accepted by M. Katori 23 January 2006     

组织型纤溶酶原激活剂及其抑制物的变化与冠心病的关系

目的：观察冠心病患者纤溶活性的变化及其在冠心病发病中的作用，探讨其临床意义。方法：用酶联免疫双抗夹心(ELISA)法测定58例冠心病患者血浆中组织型纤溶酶原激活剂(t—PA)及纤溶酶原激活抑制物-1(PAI-1)抗原含量，反映纤溶-抗纤溶活性的变化，并对冠心病患者组与对照组纤溶指标进行不同性别间的比较。结果：急性心肌梗死、不稳定心绞痛患者PM-1的含量、PAI-1／t—PA比值明显高于对照组，且急性心肌梗死患者PM-1的含量较不稳定心绞痛患者显著为高。但t—PA在急性心肌梗死、不稳定心绞痛患者均无显著降低。患者组及对照组不同性别间纤溶及抗纤溶水平均未见有显著差异。结论：血栓性疾病与纤溶系统的异常有密切关系。纤溶系统活性的变化及纤溶-抗纤溶的平衡失调在缺血性心脏病的发生、发展中起着重要作用。     

非穿透性小梁切除术治疗开角型青光眼

目的 研究非穿透性小梁切除术治疗开角型青光眼的手术效果。方法 对20例26眼开角型青光眼行非穿透性小梁切除术,术后观察眼压、结膜滤枕及前房反应等情况,随访1～12个月。结果 术后各期平均眼压均低于术前平均眼压,差异有显著性(P<0.05)术。前平均眼压26.26mmHg,术后3个月、6个月、9个月、12个月平均眼压分别为15.47mmHg、16.89mmHg、19.14mmHg、18.50mmHg。16眼结膜形成滤性手术典型滤过泡,10眼结膜疏松无滤过泡形成。26眼均无前房出血,术后前房反应轻。结论 非穿透性小梁切除术能有效地降低开角型青光眼的眼压,术后并发症少,可作为此类青光眼病人手术的选择。     

抗鼠T淋巴细胞血清的制备

目的 根据小鼠神经元与T淋巴细胞具有共同抗原Thy-1的特点，利用鼠脑组织制备兔抗鼠T淋巴细胞血清。方法 提取鼠脑组织与弗氏完全佐剂混合，制成油包水乳剂， 免疫2只家兔后，采血，分离血清，用鼠脾淋巴细胞做凝集试验及补体依赖的细胞毒试验确定其活性。结果 制备的抗血清可与鼠脾淋巴细胞发生反应，2只兔抗血清凝集试验的效价分别为1：640及1：1280。抗血清1：320倍稀释后与鼠脾淋巴细胞做补体依赖的细胞毒试验，特异性细胞毒性均为32％。结论 用鼠脑组织制备抗鼠T淋巴细胞血清是一种可行的方法。