不同来源人树突状细胞的培养诱导及鉴定

目的通过对不同来源的人树突状细胞的体外培养与诱导,建立树突状细胞的稳定体系,为更进一步实验打好基础。方法分离人外周血单个核细胞(PBMC),用不同树突状细胞亚群的特异性表面标记抗体磁珠分离树突状细胞前体,通过加入不同的刺激分子,分别诱导未成熟、成熟及活化等不同阶段的树突状细胞。结果与未成熟树突状细胞相比,成熟活化型树突状细胞表达HLA-DR、CD86以及CD40等共刺激分子明显升高,其中MoDC对成熟标记物CD83的表达水平亦较高。此外,两者在形态上也有明显的差异。结论通过对不同亚群树突状细胞的体外诱导、培养和鉴定,建立了较为成熟的研究树突状细胞的平台,为深入开展工作打下基础。     

肾移植手术中特殊供肾的处理

目的 总结特殊供肾的外科处理经验。方法 回顾性分析1996年1月-2001年6月间进行的868例尸体肾移植中326只特殊供肾的处理，并与542只普通供肾的移植效果进行比较。结果 326只特殊供肾均得到了利用，血管变异的供肾移植后1个月血肌酐水平，急性肾小管坏死（ATN）发生率及1年移植肾存活率与普通供肾均远见显著差异；其他特殊供肾均未发生与修整术有关的并发症。结论 畸形供肾或损伤供肾，通过合理手术整形，灵活运用等方法保肾，并不影响肾移植的效果。     

CD4^＋CD25^＋T细胞亚群在系统性红斑狼疮患者中的检测和意义

目的:探讨CD4+CD25+T细胞亚群在系统性红斑狼疮(SLE)发病中的作用.方法:采用流式细胞仪检测35例SLE患者(其中活动组20例)及18例健康对照组的外周血CD4+CD25+T细胞亚群的百分比及其CD4+CD25highT细胞与CD4+CD25intT细胞的比率,同时评估疾病活动指数(SLEDAI),分析两者的相关性.结果:活动组和非活动组SLE患者外周血总CD4+GD25+T细胞、CD4+CD25highT细胞和CD4+CD25intT细胞占CD4+T细胞的百分比与对照组相比,差异均无统计学意义(P>0.05),且与SLEDAI无相关性.活动组SLE患者外周血CD4+CD25highT细胞与CD4+CD25intT细胞的比率与非活动组及健康对照组相比较明显降低,差异有统计学意义(P<0.05),且与SLEDAI呈明显负相关.结论:活动性SLE患者外周血CD4+CD25highT细胞与CD4+CD25intT细胞的比率降低可能在SLE的发病中起到重要的作用.     

基于慢病毒的SHARP-2特异的RNA干扰延长肾移植受者大鼠存活时间

Objective To investigate if rat enhancer of split- and hairy-related protein-2 (SHARP-2) short hairpin RNA interference (shRNAi) prolongs the survival time of rat kidney transplant recipients. Methods Gene recombinant procedures, transfection and co-transfection were carried out to introduce short hairpin RNA interference sequences target for SHARP-2 into 3rd generation self-inactivated lentiviral-ViraPower packaging mix. Limiting dilution method was used for viral titration. Real-time PCR was employed for quantification of gene expression. Rat kidney transplantation was utilized to investigate the effect of SHARP-2 gene silence on the recipient survival. Results A lentiviral-based shRNAi construct LV-SHARP-2iC showed 84% SHARP-2 gene silence efficiency in normal rat kidney cells. At multiplicity of infection 20, 57% T cells could be transfected by lentivirus with spinoculation method. In activated T cells, SHARP-2 g ene silence resulted in 61.3% and 68.7% reduction of intedeukin 2 (IL-2) and interferon γ (IFN-γ) gene expression. When donor kidney was perfused with 5×107 TU LV-SHARP-2iC, the median survival time prolonged for 4-5 days as compared to blank and scramble control groups. Conclusions A recombinant lentivirus LV-SHARP-2iC that effectively silence SHARP-2 gene expression is constructed successfully, leading to the inhibition of IL-2 and IFN-γ. LV-SHARP-2iC treatment can prolong the survival time of rat kidney transplant recipients.     

基于慢病毒的SHARP-2特异的RNA干扰延长肾移植受者大鼠存活时间

Objective To investigate if rat enhancer of split- and hairy-related protein-2 (SHARP-2) short hairpin RNA interference (shRNAi) prolongs the survival time of rat kidney transplant recipients. Methods Gene recombinant procedures, transfection and co-transfection were carried out to introduce short hairpin RNA interference sequences target for SHARP-2 into 3rd generation self-inactivated lentiviral-ViraPower packaging mix. Limiting dilution method was used for viral titration. Real-time PCR was employed for quantification of gene expression. Rat kidney transplantation was utilized to investigate the effect of SHARP-2 gene silence on the recipient survival. Results A lentiviral-based shRNAi construct LV-SHARP-2iC showed 84% SHARP-2 gene silence efficiency in normal rat kidney cells. At multiplicity of infection 20, 57% T cells could be transfected by lentivirus with spinoculation method. In activated T cells, SHARP-2 g ene silence resulted in 61.3% and 68.7% reduction of intedeukin 2 (IL-2) and interferon γ (IFN-γ) gene expression. When donor kidney was perfused with 5×107 TU LV-SHARP-2iC, the median survival time prolonged for 4-5 days as compared to blank and scramble control groups. Conclusions A recombinant lentivirus LV-SHARP-2iC that effectively silence SHARP-2 gene expression is constructed successfully, leading to the inhibition of IL-2 and IFN-γ. LV-SHARP-2iC treatment can prolong the survival time of rat kidney transplant recipients.     

基于慢病毒的SHARP-2特异的RNA干扰延长肾移植受者大鼠存活时间

Objective To investigate if rat enhancer of split- and hairy-related protein-2 (SHARP-2) short hairpin RNA interference (shRNAi) prolongs the survival time of rat kidney transplant recipients. Methods Gene recombinant procedures, transfection and co-transfection were carried out to introduce short hairpin RNA interference sequences target for SHARP-2 into 3rd generation self-inactivated lentiviral-ViraPower packaging mix. Limiting dilution method was used for viral titration. Real-time PCR was employed for quantification of gene expression. Rat kidney transplantation was utilized to investigate the effect of SHARP-2 gene silence on the recipient survival. Results A lentiviral-based shRNAi construct LV-SHARP-2iC showed 84% SHARP-2 gene silence efficiency in normal rat kidney cells. At multiplicity of infection 20, 57% T cells could be transfected by lentivirus with spinoculation method. In activated T cells, SHARP-2 g ene silence resulted in 61.3% and 68.7% reduction of intedeukin 2 (IL-2) and interferon γ (IFN-γ) gene expression. When donor kidney was perfused with 5×107 TU LV-SHARP-2iC, the median survival time prolonged for 4-5 days as compared to blank and scramble control groups. Conclusions A recombinant lentivirus LV-SHARP-2iC that effectively silence SHARP-2 gene expression is constructed successfully, leading to the inhibition of IL-2 and IFN-γ. LV-SHARP-2iC treatment can prolong the survival time of rat kidney transplant recipients.     

居家腹膜透析并发胸腹漏12例的护理体会

总结12例居家腹膜透析患者并发胸腹漏的护理。护理重点为加强随访,对并发胸腹漏患者进行胸腔引流护理、停腹膜透析护理、小剂量腹膜透析护理及心理支持。12例患者中,6例改行血液透析,4例停腹膜透析1月行非平卧位间歇性小剂量腹膜透析,2例直接行非平卧位间歇性小剂量腹膜透析,12例患者均好转,1例已行同种异体肾移植。     

腹膜透析患者蛋白质摄入和营养状况的调查研究

众所周知,营养不良是影响腹膜透析（peritoneal dialysis,PD）患者预后的最重要的独立危险因素[1]。大量文献证明,蛋白质能量的摄入不足是引起PD患者营养不良的重要原因[2-5]。然而,美国肾脏病基金会/肾脏病临床实践指南推荐的PD患者膳食蛋白摄入量（dietary protein intake,DPI）＞1.2g/（kg·d）实际上是很难做到的。国内外研究[6-9]显示：DPI＜1.2g/（kg·d）的PD患者营养状况并不比DPI＞1.2g/（kg·d）的患者差。甚至有研究表明, DPI为0.8g/（kg·d）的PD患者也能保持良好的营养[10-11]。所以,PD患者合适的DPI一直有所争议。为探寻合适的DPI,笔者对本中心近3年内规律随访的PD患者进行了膳食蛋白质摄入和营养状况的调查和分析,现报道如下。     

细胞因子基因多态性与器官移植的关系研究进展

细胞因子和器官移植关系密切,因为外来抗原的刺激正是通过细胞因子介导而使免疫相关细胞活化,而这些活化了的细胞也正是通过细胞因子的介导才发挥了效应,导致了排斥反应。所以,凡是影响细胞因子水平的因素都有可能影响器官移植的结局。迄今为止的许多研究都表明,个体间细胞因子量存在着很大的差异,如γ干扰素(ＩＦＮγ)和白细胞介素10(ＩＬ10),高表达者和低表达者间的差异可达10倍[1]。作为细胞因子,前者是促进炎症过程的,而后者则是抵抗炎症过程的。因为排斥反应的本质其实是由一种特殊的因子启动的炎症过程,所以移植受者个体间不同细胞因…     

乙、丙型肝炎病毒感染对肾移植患者长期存活的影响

目的　了解乙型肝炎病毒（ＨＢＶ） 及丙型肝炎病毒（ＨＣＶ） 感染对肾移植患者长期存活的影响。方法　对８０ 例感染ＨＢＶ、ＨＣＶ 者肾移植术后肝病及排斥的发生情况、死亡原因及长期存活率进行分析。结果　移植后慢性肝病发生率为２１ ．２５％ , 死亡率为１８ ．７５ ％ , 显著高于非感染组（１ ．１９ ％ , Ｐ＜ ０．０１） ;ＨＣＶ 组超急性排斥及加速性排斥的发生率（６ ．０６％ ,９ ．０９ ％ ） 显著高于非感染组（０ ．７２ ％ ,２ ．７４ ％ ; Ｐ＜ ０ ．０１ , Ｐ＜ ０ ．０５）。结论　ＨＢＶ及ＨＣＶ感染显著影响肾移植受者的长期存活率; 移植后肝病及感染是其主要死因; 对ＨＢＶ 及ＨＣＶ 感染患者应采取合理的免疫抑制治疗。