目的:探讨灭活SV40致敏的猴外周血来源的树突状细胞（dendritic cells,DC）的生物学特性。方法:采用密度梯度离心法从恒河猴外周血分离出单个核细胞,然后采用玻璃黏附法分离淋巴细胞和单核细胞,使用重组人粒细胞-巨噬细胞集落刺激因子（Recombinant Human granulocyte-macrophage colony stimulating factor,rhGM-CSF）扩增单核细胞,使用重组人白细胞介素4（Recombinant Human Interleukin-4,rhIL-4）诱导单核细胞向树突状细胞方向分化,使用SV40全病毒灭活疫苗作为冲击抗原,使用重组人肿瘤坏死因子（Recombinant Human Tumor Necrosis Factor,rhTNFα）促其成熟。分别于普通光学显微镜、相差显微镜、扫描电镜下观察细胞形态。使用流式细胞术分析细胞表型特征。使用H-TdR掺人法测定混合淋巴细胞反应（mixed lymphocyte reaction,MLR）中DC刺激自体及同种异体T细胞增殖的效应。使用辣根过氧化酶（HRP）内吞实验检测其群体内吞能力。结果:使用hGM-CSF可以有效促进猴DC前体的增殖,细胞生长曲线显示,细胞增殖在第8天达到增殖高峰。使用rhIL-4可以成功诱导猴DC的分化。光镜下可见大小形态各异的树突状细胞。扫描电镜下可见吞噬SV40抗原后的形成的大量面纱DC的形态。细胞吞噬能力测定显示,细胞在第6天吞噬能力最强。MLR显示体外扩增诱导的DC能够有效刺激T细胞增殖。流式细胞术表型分析显示,表达树突状细胞CD1a特异性标志的细胞比例可达77.7%,表达树突状细胞CD83成熟标志的细胞比例可达56.5%。rhTNFα具有促进猴DC成熟的作用。结论:rhGM-CSF、rhIL-4和rhTNFα可用于猴DC的扩增、诱导和成熟刺激。灭活SV40致敏的猴外周血来源的DC在MLR中能够有效刺激T细胞增殖。体外灭活SV40抗原冲击的DC多表现为循环DC的形态。
目的 研究阿德福韦酯(adv)治疗拉米夫定(lam)耐药的慢性乙型肝炎(chb)患者的疗效和安全性.方法 治疗组32例,为lam耐药的chb患者采用adv 10 mg/d联合lam 100 mg/d治疗,疗程为48周;历史对照组24例,为lam耐药的chb患者继续服用lam 100 mg/d治疗,疗程为48周.服药后定期检测alt、hbv dna和hbeag,并观察不良反应.结果 治疗第24周和48周时,adv组hbv dna水平较用药前分别下降256 log10拷贝/ml和2.93 log10拷贝/ml,病毒学应答率分别为50.0%和75.0%,alt复常率分别为53.1%和68.8%;在48周时肝组织学改善率为65.6%,都显著高于安慰剂组(p<0.05).经统计,hbeag阴转率及血清学转换率两组比较差异无统计学意义.两组均未发现明显不良反应.结论 adv治疗lam耐药的chb安全有效.
abstract：
objective to investigate the efficacy and safety of adefovir dipivoxil (adv) in treatment of chronic hepatitis b (chb) patients with lamivudine (lam) resistance. methods there were treatment group (32 chb patients with lam resistance) and historical control group (24 chb patients with lam resistance) in this study. the treatment group received adv 10 mg/d and lam 100 mg/d for 48 weeks; the historical control group continued to use lam monotherapy. during the treatment causes, serum hbv dna levels, liver function and hbv serology were monitored regularly, and safety assessments were also conducted. results in treatment group, mean hbv dna levels decreased by 2.56 log10 eopies/ml and 2.93 log10 copies/ml, virus response rates were 50. 0% and 75.0%, alt normalization rates were 53.1% and 68.8% after 24 and 48 weeks of treatment, respectively. the histological improvement rate was 65.6% after 48 weeks. comparing with those in control group, the differences were statistically significant ( p <0. 05), while there was no significant statistical differences in hbeag loss rate and hbeag seroconversion rate between two groups. there was no severe adverse event during the treatment. conclusion adv is effective and safe in treatment of lamivudine-resistant chb.
目的 明确产超广谱β-内酰胺酶(esbls)菌株对喹诺酮类药物的耐药机制.方法 用pcr方法扩增产esbls的263株大肠埃希菌和99株肺炎克雷伯菌的qnra基因.对ctx-m和qnra基因都阳性的肺炎克雷伯菌zj96采用接合试验、southern杂交进行基因定位,用鸟枪法对分离自zj96菌株的同时含qnra和ctx-m基因的质粒(pkp96)进行全序列测定.结果 263株大肠埃希菌中有5株检出qnra基因,阳性率为1.9%;99株肺炎克雷伯菌中有8株检出qnra基因,阳性率为8.1%.zj96菌株含有ctx-m和qnra基因同时阳性的大小约60 kb的接合性质粒pkp96.菌株zj96的接合菌质粒pkp96含有qnra、ctx-m-24、aac(6')-ib-cr、teta和intⅠ 1等基因.结论 喹诺酮耐药基因qnra和ctx-m-24型esbls基因同时位于可接合性质粒中,耐药质粒的广泛传播是造成临床耐药菌株大量出现的主要原因.
abstract：
objective to characterize the prevalence of plasmid-mediated quinolone resistance determinants qnra in extended-spectrum β-lactamase(esbl)-producing escherichia coli and klebsiella *** pcr was used to amplify qnra gene in esbl-rpoducing isolates(including 263isolates of escherichia coli and 99 isolates of klebsiella pneumonia).conjugation experiments and southern blot hybridization were employed to definitude the location of the genes in zj96 isolate of klebsiella pneumonia which had positive qnra and ctx-m *** gun sequencing was performed for analyzing the complete nucleotide sequence of pkp96,a plasmid containing qnra and ctx-m-24 genes in zj96 *** qnra was detected in 5 out of 263(1.9%)escherichia coli isolates and 8 out of 99(8.1%)klebsiella pneumonia ***96,a conjugative plasmid including qnra gene and ctx-m-24 gene presented in zj96 *** sequence of the plasmid pkp96 displayed the qnra,ctx-m-24,aac(6')-ib-cr,teta and int Ⅰ 1 *** the plasmid-mediated genes,such as qnra and ctx-m,may facilitate the prevalence of multi-drug resistant strains.
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