变性高效液相色谱对CTX-M型超广谱β内酰胺酶基因分型研究

Objective To investigate the genotype distribution of CTX-M-type extended-spectrum β-1actamase(ESBLs) by denaturing high-performance liquid chromatography(DHPLC) in Hunan Province and the accuracy of DHPLC assay. Methods The blaCTX-M genes of standard strains and clinical ESBLs-producing Enterobacteriaceae were amplified by multiplex PCR followed by DHPLC and genotype determination. 25 isolates randomly selected were sequenced to assess the accuracy of DHPLC method. Results Among 142 ESBLs-producing isolates, 109 isolates carried blaCTX-M gene (76. 8% ). Four different CTX-M genotypes were detected by DHPLC, including CTX-M-3 (33 isolates), CTX-M-15 (19 isolates), CTX-M-14 (52 isolates) and CTX-M-9 (5 isolates). The DHPLC typing of 25 isolates suggested that 24 isolates were verified uniformly by the sequencing, but one CTX-M-15 isolate typed by DHPLC was shown to be CTX-M-82 by sequencing. Conclusion DHPLC is a powerful tool for genotyping of the resistance gene and is worth being applied in the clinical and scientific research with accurate, rapid and economic advantages.     

细胞因子介导的人脐血单个核细胞体外扩增并重建NOD/SCID小鼠造血及免疫功能

背景:细胞因子介导的脐血造血细胞体外扩增有望能解决脐血移植数量不足的问题.目的:实验拟确立在无基质培养条件下体外扩增脐血单个核细胞最合适的细胞因子组合及干细胞因子、FLT3配基协同促聚核细胞生成因子对造血及免疫重建功能的影响.方法:将新鲜脐血标本分离的单个核细胞接种于含有不同细胞因子组合的无血清无基质培养体系中培养7 d,根据不同细胞因子组合分组,将3因子干细胞因子+FLT3配基+促聚核细胞生成因子组合在无血清无基质条件下扩增培养7 d前后的脐血单个核细胞移植给经亚致死量照射的NOD/SCID小鼠.在扩增培养0,7 d检测脐血CD34+细胞数及CD34+CXCR4+,CD34+CD62L+,CD34+CD49d+的细胞数.脐血移植6周后通过流式细胞仪,PCR法检测存活小鼠体内的人源性细胞.结果与结论:移植6周后,存活小鼠体内均可检测到人源性CD45+细胞,扩增脐血移植组的NOD/SCID小鼠存活率和人特异性基因捡出率均高于新鲜脐血移植组和高于生理盐水移植组(P<0.05).扩增脐血组存活NOD/SCID小鼠骨髓中可检测到人髓系细胞(CD33+),T淋巴细胞(CD4+),B淋巴细胞(CD19+)和人造血干细胞成分(CD34+)细胞的表达.结果提示干细胞因子+FLT3配基+促聚核细胞生成因子3因子组合脐血造血细胞体外扩增是最合适的细胞因子组合,其扩增的脐血单个核细胞能够植入并重建NOD/SCID小鼠的造血及免疫功能.     

骨髓基质细胞对多细胞因子介导人脐血单个核细胞体外扩增的影响

背景:体外扩增脐血造血细胞的目的是促进脐血造血细胞的植入能力,细胞因子介导的脐血造血细胞能使细胞数有效扩增,但同时也耗竭了具有分化潜能的造血干细胞.目的:观察骨髓基质细胞对多细胞因子组合介导人脐血单个核细胞体外扩增及扩增后黏附分子和CXCR4表达的影响.方法:将分离的人脐血单个核细胞接种在预先建立的人骨髓基质细胞层上,分组培养:对照组仅含有脐血单个核细胞;多种细胞因子+脐血单个核细胞组;骨髓基质细胞+脐血单个核细胞组;骨髓基质细胞+多种细胞因子+脐血单个核细胞组.培养0,7 d检测有核细胞数, CD34+细胞数及CD34+CXCR4+,CD34+CD62L+,CD34+CD49d+的细胞数.结果与结论:单纯骨髓基质细胞和单用细胞因子组均可有效地扩增脐血造血细胞,并提高造血细胞上CD49d,CD62L及 CXCR4表达.而单用细胞因子组促进脐血造血细胞扩增能力较单纯骨髓基质细胞强,提高造血细胞CD49d,CD62L及 CXCR4表达能力较单纯骨髓基质细胞差.提示骨髓基质细胞虽可支持脐血造血细胞扩增,但难以实现造血细胞的大量扩增,但在骨髓基质细胞层的支持下,多细胞因子可有效地促进脐血造血细胞的扩增,并优于单用细胞因子及骨髓基质细胞,证实骨髓基质细胞可协同多细胞因子有效地促进脐血单个核细胞的扩增,促进造血干细胞的黏附和趋化能力.     

变性高效液相色谱对CTX-M型超广谱β内酰胺酶基因分型研究

Objective To investigate the genotype distribution of CTX-M-type extended-spectrum β-1actamase(ESBLs) by denaturing high-performance liquid chromatography(DHPLC) in Hunan Province and the accuracy of DHPLC assay. Methods The blaCTX-M genes of standard strains and clinical ESBLs-producing Enterobacteriaceae were amplified by multiplex PCR followed by DHPLC and genotype determination. 25 isolates randomly selected were sequenced to assess the accuracy of DHPLC method. Results Among 142 ESBLs-producing isolates, 109 isolates carried blaCTX-M gene (76. 8% ). Four different CTX-M genotypes were detected by DHPLC, including CTX-M-3 (33 isolates), CTX-M-15 (19 isolates), CTX-M-14 (52 isolates) and CTX-M-9 (5 isolates). The DHPLC typing of 25 isolates suggested that 24 isolates were verified uniformly by the sequencing, but one CTX-M-15 isolate typed by DHPLC was shown to be CTX-M-82 by sequencing. Conclusion DHPLC is a powerful tool for genotyping of the resistance gene and is worth being applied in the clinical and scientific research with accurate, rapid and economic advantages.     

利妥昔单抗鞘内注射治疗原发中枢神经系统淋巴瘤1例并文献分析

1病例介绍 患者，男性，44岁，因反复头痛20余天，伴行走不稳1周于2008年2月10日入院我院神经外科。入院前在当地行头颅CT、检查发现头颅占位。入院时查体：生命征正常，神志清楚，意识清晰，对答切题。四肢肌力及肌张力正常。心肺腹无异常。生理反射存在，病理反射未引出。指鼻试验、快速轮替试验、闭目难立加强试验、直线行走试验均阳性。入院后行头颅MRI检查结果：右侧额叶白质、基底节、肼胝体膝部异常信号灶及右侧小脑半球异常信号灶。     

重组甘精胰岛素联合阿卡波糖对新诊断的2型糖尿病患者β细胞功能改善的自身对照研究

目的探讨短期（3个月）重组甘精胰岛素＋三餐阿卡波糖强化治疗对新诊断的T2DM患者胰岛口细胞功能及血糖控制的影响。方法采用自身前后对照的方法,对60例新诊断但未治疗的T2DM患者给予3个月重组甘精胰岛素＋阿卡波糖强化治疗,治疗前后分别行静脉葡萄糖耐量试验（IVGTT）,计算0-10min时胰岛素曲线下面积（AUC0-10）、1～3min急性胰岛素对葡萄糖的分泌反应（AIR1-3）、胰岛素分泌指数（HOMA-β）、胰岛素抵抗指数（HOMA-IR）的变化。结果治疗后IⅥHT试验的AUC0-10、AIR1-3。及HOMA-β较试验前明显增加,而空腹及餐后血糖、HbA1c、HOMA-IR较治疗前明显下降。结论新诊断的T2DM患者接受短期甘精胰岛素＋阿卡波糖治疗使血糖快速、安全达标的同时显著改善胰岛β细胞功能和胰岛素敏感性,治疗简便,依从性较高。     

不同来源血清和细胞因子对骨髓基质细胞成纤维细胞集落形成的影响

目的观察sD大鼠骨髓基质细胞（BMSC）体外培养的生物学特性并加以鉴定,并探讨不同来源血清和细胞因子对其成纤维系祖细胞（CFU-F）产率的影响。方法取SD大鼠骨髓,进行BMSC原代培养。传代后分别观察其生长特性,绘制贴壁率,测定生长曲线,并应用流式细胞仪鉴定细胞表型;采用体外长期液体培养法观察CFU—F的产率。结果原代培养的BMSCs生长良好,倍增时间约为40小时,传代后12小时贴壁率达85％以上;胎牛血清较脐血清和马血清能促进CFU-F的增值和分化。与未加细胞因子对照组相比,单用细胞因子SCF,SDF-1均可促进CFU—F的增殖,但二者之间差异无显著性（P〉0．05）,同时加入SCF,SDF-1可明显的促进CFU-F的增殖,并优于单用SCF和SDF-1（P〈0．05）。结论体外培养的BMSCs生长稳定,传代后细胞适应性强;胎牛血清中可能较脐血清和马血清存在较多的对CFU—F有促进作用的生长因子;SCF可协同SDF-1促进CFU-F的增殖。