麻风病

20060098 建立巢式PCR和异源双链法检测石蜡标本麻风菌DDS耐药株；20060099 新发和复发麻风患中麻风菌的氨苯砜、利福平耐药基因的检测；20060100 甘肃省麻风病人血缘性与非血缘性亲属家庭发病的对比研究；……[编按]     

中国汉坦病毒H82O5株G2糖蛋白基因的克隆及在真核细胞中的瞬时表达

从感染病毒乳鼠脑组织提取总RNA，采用RT－PCR和分子克隆技术将扩增到的G2糖蛋白基因插入含CMV启动子的pcDNA3.1/His质粒载体中，通过脂质体介导转染COS－7细胞，用SDS－PAGE、Western-blot及IFIA方法分别测定表达产物的相对分子量及特异性。结果证明获得正向插入的G2－pcDNA3.1/His重组表达质粒，表达产物的相对分子量为56ku，与理论预期大小一致，并且可与汉坦病毒H8205株的腹水抗体起特异反应。表明构建的G2－pcDNA3.1/His重组质粒所表达的蛋白为中国汉坦病毒株特有，能在哺乳动物细胞中表达并具有抗原性，重组质粒可应用于汉坦病毒的DNA疫苗研究。     

新的“芯片”可以快速检测禽流感病毒

据Medscape．com 11月8日报道，美国研究人员指出，一种新的“芯片”可以检测11种不同的流感病毒株，包括禽流感病毒。在15分钟之内可以诊断出流感患。该项研究仍在进行中，研究人员希望把它发展成能够现场检测流感病毒的技术。     

马蜂毒对癌瘤细胞杀伤作用的实验研究

马蜂毒确有直接杀伤肿瘤细胞的作用，其机理可能由于马蜂具有抑制细胞分裂作用，肿瘤细胞不能分裂增殖而受到抑制。     

幽门螺杆菌粘附作用的体外研究

置幽门螺杆菌(HP)悬液于含人喉癌上皮细胞(Hep-2)或人胃癌上皮细胞(MKN-28)的培养瓶内,经2h或3.5h培养后观察HP对细胞的粘附情况。结果发现HP能成丛地粘附在上皮细胞表面,少数侵入胞浆内。HP对MKN-28粘附率明显高于对Hep-2细胞者。随菌龄的延长或菌株的变异,HP的粘附作用明显减弱。1%D-甘露糖对HP的粘附性有部分阻抑作用。     

鸡柔嫩艾美耳球虫疫苗候选基因在大肠杆菌中的表达

目的构建柔嫩艾美耳球虫子孢子表面抗原原核表达载体，并且在大肠杆菌中表达。方法将本室构建的pMD-Mz5-7克隆质粒酶切回收目的片段定向亚克隆到pET-28b( )载体上，构建成原核表达载体pET-28b-Mz5-7，酶切鉴定正确后，在Escherichia coli BL21(DE3)中用IPTG诱导表达，并经SDS-PAGE及Western blotting鉴定。结果成功构建了目的抗原基因的原核重组表达质粒pET-28b-Mz5-7，IPTG诱导表达该融合蛋白，SDS-PAGE电泳表明，其能表达一分子质量约为37ku的融合蛋白，与预测分子质量相符，最佳反应条件为1mol／LIPTG诱导4h后表达量最高。薄层扫描显示表达的融合蛋白占菌体蛋白总量的18％，Western blotting结果显示，表达产物可被抗柔嫩艾美尔球虫的多克隆抗体识别，说明该融合蛋白具有很好的反应原性。结论在原核细胞融合表达Mz5-7成功，为鸡球虫病重组苗的研究奠定了基础。     

NK／LAK细胞对人口腔癌耐细胞药株杀伤活性的观察

OBJECTIVE: To understand the relation between cytotoxic activity of immunologic effector cells and multidrug resistance of the tumor cells. METHODS: Continuous observation of the morphological changes and MTT colorimetry were employed to evaluate the cytotoxic activity of lymphokine-activated killer (LAK) cells and natural killer (NK) cells against multidrug-resistant (MDR) human oral carcinoma cell line-KBV200 (before and after reversal of MDR) and parental drug-sensitive cell line KB. The morphologic changes of LAK cells and the 3 target cell lines were observed continuously under inverted microscope 3 h after co-culture of LAK cells with one of three target cell lines respectively. The lysis rates of three tumor cell lines in response to co-culture with LAK or NK cells were determined using MTT colorimetry. RESULTS: In comparison with the parental drug-sensitive cell line KB, both KBV200 and its reserved cell line by verapamil (KBVV) showed earlier adherence and greater number of cells lysed by LAK. In MTT colorimetry assay, the cytotoxicity of both LAK and NK cells against the 3 cell lines was associated with the effector-to-target (E/T) cell ratio; the lysis rates of KBV200 and reversed KBV200 cells by verapamil in response to LAK and NK cells were higher than that of KB cells (P<0.05), but KBV200 and KBVV did not significantly differ (P>0.05). At the same E/T ratio, LAK cells possessed stronger cytotoxicity than NK cells against all the tumor cell lines (P<0.05). CONCLUSIONS: Immunologic effector cells possess strong cytotoxic activity against multidrug-resistant cell line KBV200. Modulation of MDR does not decrease the cytotoxic activity of the immunologic effector cells. The results of this study suggest that adoptive cell immunotherapy with immunologic effector cells may be of value in controlling the progress of MDR tumors.     

真菌病

20 0 4 0 779　唑类药物体外诱导白念珠菌耐药的初步研究/宋晓静(浙大一附院皮肤科)…∥临床皮肤科杂志.- 2 0 0 3,32 (10 ) .- 575～576将质控株及敏感白念珠菌在含不同抑菌浓度的氟康唑(FCZ)、酮康唑(K CZ)、伊曲康唑(ICZ)的液体培养基中进行人工传代诱导耐药。结果白念珠菌在高浓度的唑类药物培养基中能诱导耐药,且为交叉耐药,而在低浓度中经传2 0代仍不能诱导耐药,诱导后的耐药菌株在无药物培养基中传代可恢复敏感性。提示唑类药物能诱导耐药白念珠菌的产生,但与诱导药物的浓度有关,且为不稳定耐药。(汤亚娥)2 0 0 4 0 780　白念珠…