舍曲林抗新生隐球菌的体外及动物实验研究

目的评估舍曲林抗新生隐球菌的效果。方法实验分为6组,分别为空白对照组、10 mg/mL氟康唑、10 mg/mL舍曲林、20 mg/mL舍曲林、10 mg/mL氟康唑联用10 mg/mL舍曲林以及10 mg/mL氟康唑联用20 mg/mL舍曲林组。通过体外药敏试验及BALBc小鼠新生隐球菌动物探讨各组间抗隐球菌效果的差异。结果体外药敏试验发现舍曲林可有效降低新生隐球菌菌落数,当与氟康唑联用时抑菌效果更显著。动物实验发现2种浓度的舍曲林都可明显降低感染小鼠实验早期脑、肺组织的新生隐球菌菌落数,但在实验后期,低浓度的舍曲林对感染小鼠脑组织失去抑菌作用。脑、肺组织中,舍曲林治疗对新生隐球菌的抗菌效果均不如氟康唑。舍曲林与氟康唑联合用药对新生隐球菌模型小鼠肺组织的抗菌效果强于单用舍曲林或氟康唑。结论舍曲林具有抗新生隐球菌的作用,当与氟康唑联合用药时可起到协同作用。     

聚丙烯酰胺凝胶碎块吸收洗脱法浓缩病毒的实验研究及应用

本文报告聚丙烯酰胺凝胶(PAG)碎块吸收洗脱法在浓缩新城鸡瘟病毒(NDV)和人轮状病毒(HRV)方面的实验研究及应用。我们通过一次和二次浓缩,分别将NDV感染的鸡胚尿液稀释液浓缩了12.1倍和90.2倍,血凝滴度相应增高,总病毒回收率分别达到90.2％84.9％。用二次浓缩后的含HRV粪便上清液进行RNA PAGE银染色检测,获得了HRV RNA电泳型检测阳性稀释度提高32倍左右的结果。据此,采用这一方法,对临床RNA PAGE检测阴性的33例婴幼儿腹泻粪便进行PAG浓缩处理后再检测,发现有12例转为阳性,阳转率为36.4％。从而,使HRV总检出率由67％增至79％。     

凝胶电泳分析蛋白酶活性的技术改进

蛋白酶在植物生长代谢过程中具有重要的生理作用。本文探讨了利用“凝胶电泳技术”分析蛋白酶活性的改进方法 ,通过改变电泳分离胶中底物蛋白浓度估算了“目标蛋白酶”分子量 ;通过改变电泳前的样品处理条件了解蛋白酶的部分特性 ;采用分离胶切割法了解“目标蛋白酶”的最适反应温度范围、最适反应pH范围及蛋白酶的类型等。并对影响凝胶电泳蛋白酶活性检测结果的各种因素作了分析     

结核分枝杆菌感染实验模型

结核分枝杆菌是引起人结核病的主要病原,全世界约有1/3人口感染结核分枝杆菌。尽管该病原可感染并引起许多动物疾病,但人类是其中心宿主。为研究结核分枝杆菌的致病机理及宿主对本病原的保护性和免疫病理学反应,选择合适的动物模型非常必要。本文阐述了结核病研究中常用的实验模型及各种模型的优缺点。实验模型的合理应用将促进我们对结核病的认识,从中获取的资料将有助于我们发现更好的预防和治疗方案。     

Activity of 7-methyljuglone derivatives against Mycobacterium tuberculosis and as subversive substrates for mycothiol disulfide reductase

The naphthoquinone 7-methyljuglone (5-hydroxy-7-methyl-1,4-naphthoquinone) has previously been isolated and identified as an active component of root extracts of Euclea natalensis which displays antitubercular activity. Herein, a series of synthetic and plant-derived naphthoquinone derivates of the 7-methyljuglone scaffold have been prepared and evaluated for antibacterial activity against Mycobacterium tuberculosis. Several of these compounds have been shown to operate as subversive substrates with mycothiol disulfide reductase. The absence of a direct correlation between antitubercular activity and subversive substrate efficiency with mycothiol disulfide reductase, might be a consequence of their non-specific reactivity with multiple biological targets (e.g. other disulfide reductases).     

The Incidence of Non-tuberculous Mycobacterium Lung Disease in Patients with Suspected Pulmonary Tuberculosis

Non-tuberculous mycobacterium (NTM) lung disease is increasing in prevalence. We analyzed the frequency of NTM lung disease among patients who are suspected of tuberculosis. NTM was isolated from about one-fourth of the mycobacterium culture-positive patients and about half of these had NTM lung disease. Therefore, NTM isolates should be routinely identified at the species level for adequate treatment.     

Synthesis of trehalose-based compounds and their inhibitory activities against Mycobacterium smegmatis

The synthesis of a library of trehalose-based compounds has been accomplished, and their activities against Mycobacterium smegmatis have been determined. A preliminary structure–activity relationship (SAR) is reported. Despite not having a potent lead, one of the trehalose derivatives displays strong activity when applied with isoniazid (INH), which is known to have low sterilizing activity. The bacteriocidal nature of our compounds against Mycobacterium may be significant for the development of new therapies against tuberculosis.     

随机扩增多态性DNA技术对结核分支杆菌菌株鉴别及其耐药相关性分析

用引物G1和L1扩增的16S-23SrDNA间隔区作为模板DNA,再用引物AP50进行随机扩增多态性DNA（Random Amplified PolymorphicDNA,RAPD)分棉线AP50扩增出的各菌株DNA片段,用1.2%的琼脂糖凝胶电泳EB染色,可获得1或2个大小不同的DNA片段（390-1031bp)。各株间差异明显,结果表明,不同菌株有不同的DNA多态性,药物敏感株与耐药株之间多态性差异明显,耐药株中耐药谱相似或相同的菌株间其多态性也相似,RAPD有助于简便,快速,有效地了解菌株间的克隆相关性及耐药性传播。     

肺上皮细胞自噬体在结核分枝杆菌感染中保护作用的分子机制

目的：构建Atg5．真核表达载体并瞬时转染肺上皮细胞细胞株,探讨自噬在结核分枝杆菌感染上皮细胞中保护作用的分子机制。方法：设计针对Atg5的RNAi序列,化学合成后经过变性,退火连接到pSilencerTM3．1-H1hygro真核表达载体,经测序验证其正确性。脂质体法瞬时转染真核细胞A549,免疫印迹法检测瞬时转染的效果。用结核分枝杆菌分别感染正常和自噬表达低下的A549细胞．通过检测LDH来观察细胞的坏死情况。结果：成功的构建了pSilencerTM3．1-H1hygro真核表达载体并建瞬时转染了A549细胞株,成功抑制了细胞的自噬功能。Atg5-细胞对结核杆菌的抵抗能力下降。结论：在自噬表达低下的细胞中,细胞对结核分枝杆菌的抵抗能力有明显下降。在结核分枝杆菌感染上皮细胞的过程中,自噬是一种保护机制。     

Rpf结构域及其突变体对MTB感染小鼠的免疫治疗作用

目的:观察复苏促生长因子Rpf结构域(Rpfd)及其突变体E54K (Rpfd1),E54A (Rpfd2)和E54K+ D48A (Rpfd3)对MTB感染小鼠的免疫治疗作用.方法:表达、纯化Rpfd及其突变体Rpfd1,Rpfd2和Rpfd3.结核毒株H37RV从尾静脉感染BALB/c小鼠,剂量为1× 105C F U.感染4w后,随机分为6组,在感染后4w、6w和8周分别腹部皮下注射含50 μg/ml相应蛋白Rpfd、Rpfd1、Rpfd2、Rpfd3的生理盐水,同时设异烟肼(INH,54.25 mg/L)联合利福平(RFP,52.5 mg/L)治疗组.在感染后11w和13w,分别取一侧肺脏计数荷菌数.将各时间点的感染小鼠另一侧肺脏固定后进行组织学观察.结果:表达产物均可与抗-Rpf结构域单抗特异性结合,相对分子量约为32 kDa.感染后11w和13w,Rpfd、Rp fd1、Rpfd2治疗组肺部荷菌量明显少于Rpfd3治疗组和生理盐水组(P＜0.05),但其对肺部MTB的控制未达到INH+RFP联合治疗的效果(P＜0.05).结论:MTB感染小鼠后,利用Rpfd蛋白进行免疫治疗,发现Rpfd及其突变体Rpfd1和Rpfd2蛋白能够显著的降低肺脏荷菌数,并且诱导淋巴细胞的增值.为利用Rpf结构域突变体制备MTB的免疫治疗疫苗提供了理论依据和实验基础.     

Biology of IL-27 and its Role in the Host Immunity against Mycobacterium Tuberculosis

IL-27, a heterodimeric cytokine of IL-12 family, regulates both innate and adaptive immunity largely via Jak-Stat signaling. IL-27 can induce IFN-γ and inflammatory mediators from T lymphocytes and innate immune cells. IL-27 has unique anti-inflammatory properties via both Tr1 cells dependent and independent mechanisms. Here the role and biology of IL-27 in innate and adaptive immunity are summarized, with special interest with immunity against Mycobacterium tuberculosis.