基因诊断技术在脊柱结核诊断中的应用现状

脊柱结核是一种常见的肺外结核病。近年来,随着脊柱结核患者逐年增多,不典型脊柱结核患者亦逐渐增多,而不典型脊柱结核诊断较为困难,需要与一般细菌感染、肿瘤以及非结核分枝杆菌感染相鉴别。基因诊断技术是一项诊断脊柱结核的重要工具,对临床不典型脊柱结核的诊断有较高价值。本文将对脊柱结核的流行现状、诊断及基因诊断技术进行综述,以帮助临床工作者对脊柱结核做出更加准确的诊断。     

银离子、铜离子和锌离子对常见念珠菌的抗菌性能比较

目的 探讨银离子(Ag⁺)、铜离子(Cu²⁺)、锌离子(Zn²⁺)对5种常见念珠菌(白色念珠菌、热带念珠菌、近平滑念珠菌、克柔念珠菌和光滑念珠菌)的抗菌作用。方法 使用去离子水分别将硝酸银、氯化铜和氯化锌配制为0.1 mol·L^-1的水溶液。取保存的白色念珠菌标准菌株ATCC-60193、近平滑念珠菌、光滑念珠菌、热带念珠菌、克柔念珠菌,加入溶菌肉汤培养液,使用比浊仪调配菌液浊度为0.5麦氏单位,然后使用无菌去离子水将菌液稀释至10⁶ CFU·L^-1。取配制好的0.1 mol·L^-1的硝酸银、氯化铜和氯化锌溶液,分别倍比稀释为6个浓度(10^-2、10^-3、10^-4、10^-5、10^-6、10^-7 mol·L^-1),分别取各浓度的金属离子溶液100μL与预制好的5种念珠菌悬液100μL进...     

胰腺实性-假乳头状肿瘤内PTEN和nestin的表达

目的 探讨胰腺实体假乳头肿瘤（SPTP）细胞可能的组织学起源。方法 复习25例SPTP的临床病理特征，常规HE和超微结构观察，免疫组化EnVision法染色检测肿瘤细胞PTEN和nestin等的表达。结果 25例SPTP中女性22例，男性3例，中位年龄22．3岁。肿瘤主要位于胰腺内，1例位于后腹膜并与胰腺相连。1例伴有肝转移。肿瘤呈囊实性，出血、坏死明显。肿瘤细胞大小形态较一致，实性和假乳头状排列，部分似“室管膜样”菊形团。核卵圆形，有核沟，核仁不明显，核分裂象少见。肿瘤细胞均有PTEN阳性表达（18／18，100％），44％（8／18）的病例有nestin的表达。超微结构观察细胞内可见酶原样分泌颗粒和神经内分泌颗粒。结论 胰腺实体假乳头肿瘤可能起源于胰腺多能干细胞。     

Triptolide inhibits IL-12 production and T cell-stimulatory capacity of dendritic cells

Extracts of Tripterygium wilfordii Hook F.(TWHF ) are effective in traditional Chinesemedicine for treatment of autoimmune diseasessuch as rheumatoid arthritis, systemic lupus ery thematosus, nephritis and asthma [1, 2]. Trip tolide, a diterpenoid triepoxide, is derived fromTWHF and is responsible for most of the immuno suppressive and anti inflammatory effects ofTWHF. Triptolide has been shown to be effectivein the treatment of autoimmune diseases, …     

The Distribution of Synaptotagmin II in RBL-2H3 and Its Regulation on Exocytosis of Lysosomes in RBL-2H3

Synaptotagmin(Syt)constitutes a family of membrane-trafficking proteins,so far nearly 20 Syts have beendiscovered.Extensive work showed that synatotagmins were a potential Ca~(2+) sensor for regulated exocytosis.Thisstudy was to investigate the expression and location of synaptotagmin Ⅱ(Syt2)in RBL-2H3(RBL)and its role inregulating exocytosis of RBL.The expression of Syt2 in RBL was confirmed by Western blot.The recombinantexpression vector pEGFP-N1-Syt2 was constructed and transfected into RBL by electroporation,the stabletransfectant RBL-Syt2-S expressing fusion protein Syt2-EGFP were obtained and Syt2 was highly concentrated atplasma membrane with little detected in cytoplasm.To analyze the role of Syt2 during exocytosis of RBL,therelease of cathepsin D was assayed by immunoblotting.Compared with control,the release of cathepsin D byRBL-Syt2-S was markedly decreased.The results indicated that Syt2 played a negative regulation in exocytosis oflysosomes in RBL.Cellular & Molecular Immunology.2005;2(3):205-209.     

Plasma HIV-1 RNA decline within the first two weeks of treatment is comparable for nevirapine, efavirenz, or both drugs combined and is not predictive of long-term virologic efficacy: A 2NN substudy

BACKGROUND: The initial rate of plasma HIV-1 RNA (pVL) decline has been proposed as a marker of early efficacy of antiretroviral therapy (ART) and a possible predictor of late efficacy. We compared the rate of pVL decline in patients starting ART with nevirapine (NVP), efavirenz (EFV), or both drugs combined in addition to lamivudine (3TC) and stavudine (d4T). METHODS: Analysis of the viral decay constant (VDc) during the first 2 weeks of treatment in patients enrolled in the 2NN study who remained on allocated treatment. RESULTS: The median VDc (log10 copies per day, [interquartile range]) was similar for NVP (0.30 [0.25-0.36], EFV (0.31 [0.27-0.37]), and NVP + EFV (0.30 [0.27-0.36]). Patients with a baseline pVL >100,000 copies/mL were 8.7 (95% confidence interval [CI]: 6.2-12.3) times more likely to have a VDc >75th percentile. A high VDc was not associated with plasma drug concentration or with a decreased risk of virologic failure at week 48 after the start of therapy (hazard ratio = 0.8, 95% CI: 0.6-1.2). CONCLUSION: NVP, EFV, or NVP + EFV in combination with 3TC and d4T show similar rates of pVL decline during the first 2 weeks of treatment. The VDc with these regimens is not predictive of late virologic efficacy.     

突变型K5重组蛋白抑制小鼠肝癌血管生成和肿瘤生长的作用

目的：体内实验观察人纤溶酶原K5缺失突变体Ⅰ（K5 mut1）对HepA小鼠肝癌血管生成和肿瘤生长的影响。 方法:大肠杆菌中表达，组氨酸结合柱亲和层析、纯化获得K5 mut1蛋白，SDS-PAGE和Western blotting方法鉴定其表达；建立皮下种植肝癌小鼠模型，腹腔注射不同剂量K5 mut1重组蛋白，检测抑瘤率及肝癌组织微血管密度（MVD）。结果： SDS-PAGE及Western blotting鉴定获得K5 mut1纯化蛋白；K5 mut1剂量依赖性地抑制小鼠肝癌（HepA）实体瘤生长，不同剂量K5 mut1治疗组肝癌组织MVD低于对照组，并随K5 mut1用药剂量增加而降低。 结论： K5 mut1具有抑制小鼠肝癌生长的作用，抑制肿瘤血管生成可能是K5 mut1抑制肿瘤生长的主要机制。结果提示K5 mut1具有治疗肝癌的潜在临床价值。     

Human neutrophils secrete gelatinase B in vitro and in vivo in response to endotoxin and proinflammatory mediators

Bacterial sepsis is characterized by a systemic inflammatory state, with activation of numerous cell types. Phagocytes participate in this phenomenon by secreting various proinflammatory cytokines and enzymes. Matrix metalloproteinases (MMPs) such as gelatinases are produced by phagocytes and are thought to play an important role in processes of cell transmigration and tissue remodeling. In this work, we show that endotoxin (lipopolysaccharide [LPS]) and other inflammatory mediators, such as tumor necrosis factor (TNF), interleukin-8, and granulocyte colony-stimulating factor, induce a rapid (within 20 min) release of gelatinase-B (MMP-9) zymogen in whole human blood, as determined by gelatin zymography. The polymorphonuclear neutrophil was identified as the cell responsible for this rapid secretion, as a result of the release of preformed enzymes stored in granules. Normal human subjects given LPS intravenously showed a similar pattern of proMMP-9 secretion, with maximum plasma levels reached 1.5 to 3 h after LPS administration (P = 0.0009). Prior administration of TNF receptor:Fc, a potent TNF antagonist, to subjects given LPS, only partially blunted the release of proMMP-9 (P = 0.033). Ibuprofen, a cyclooxygenase inhibitor, did not alter this pattern of release. Increased levels of proMMP-9 and proMMP-2, as well as activated forms of MMP-9, were found in plasma from two patients with gram-negative sepsis. The levels of MMPs paralleled the severity of clinical condition and a marker of the severity of sepsis, plasma procalcitonin. These data indicate that MMPs are released in whole blood in response to various inflammatory mediators and that they could serve as sensitive and early markers for cell activation during the course of bacterial sepsis.     

路氏乳杆菌JCM1081黏附相关蛋白的初步鉴定

目的研究路氏乳杆菌(Lactobacillus reuteri,也称罗伊氏乳杆菌)JCM1081菌体表面蛋白对其黏附HT-29细胞的影响。方法将路氏乳杆菌JCM1081菌体进行胰蛋白酶、蛋白酶K处理;用氯化锂和盐酸胍对乳杆菌表面的蛋白进行抽提,进行SDS-PAGE后与黏蛋白受体进行Western blot,并对杂交阳性蛋白进行质谱分析鉴定。结果路氏乳杆菌JCM1081菌体经胰蛋白酶、蛋白酶K处理后,其对HT-29细胞的黏附力显著下降(P<0．01);用氯化锂去除路氏乳杆菌JCM1081菌体外表面的S层蛋白后,路氏乳杆菌JCM1081对HT-29细胞的黏附力无显著变化;Western blot结果显示相对分子质量(Mr)为29×103和14×103的两种菌体表面蛋白与黏蛋白受体杂交中出现了强阳性;质谱分析结果显示29×103蛋白与路氏乳杆菌ATCC55730的h0793蛋白相似性高达71．1％。结论路氏乳杆菌JCM1081菌体表面的蛋白参与了乳杆菌的黏附,其中29×103和14×103的两种胞壁表面蛋白能够特异地识别黏蛋白受体并与之结合,29×103蛋白属ABC转运蛋白家族。