结核分枝杆菌Ag85B/GST融合蛋白表达载体构建及在大肠杆菌中的表达

目的 构建结核分枝杆菌Ag85B／GSF融合蛋白表达质粒，并在大肠杆菌(E．coli)中诱导表达。方法 以质粒pUC18／Ag85B为模板，用PCR法扩增结核杆菌Ag85B基因，扩增的Ag85B上游含有EcoR Ⅰ酶切位点，下游含有SalⅠ酶切位点；将Ag85B基因定向插入质粒pGEX-4T-1中，构建原核表达质粒pGEX-Ag85B并转化E．coli B121，筛选阳性重组子，限制性内切酶切鉴定和DNA序列测定，异丙基硫代半乳糖苷(IPTG)诱导表达，SDS-PAGE和Westem blot鉴定结果。结果 成功构建原核表达质粒pGEX-Ag85B并表达出Mr约56000大小的Ag85B／GSY融合蛋白，表达量占总菌体的30％。结论 为进一步进行纯化Ag85B蛋白和研究其在膀胱肿瘤治疗中的作用奠定了基础。     

抗结核药物所致皮疹临床特点及停药指征的探讨

目的 探讨抗结核药物所致皮疹的临床特点及停药指征.方法 对1998-2005年收治的3 082例住院结核病人中,因抗结核药物所致皮疹296例病人进行分析.结果 变态反应总发生率为10.3%,皮疹发生率为9.6%（296例）,其中单纯皮疹88例,占2.9%,皮疹伴其他系统表现一种或一种以上者208例,占6.7%;无皮疹者22例,占0.7%.皮疹病例总停药率为61.5%.结论 抗结核药物所致过敏反应多为全身性,发生过敏反应后,绝大多数单纯皮疹病人经过抗过敏处理后仍可维持原方案继续治疗,而皮疹伴有明显肝功能异常、心律失常、关节痛和过敏性休克者均需立即停用抗结核药物,并且避免再次使用.     

左氧氟沙星治疗HBV携带肺结核患者疗效观察

目的:探讨左氧氟沙星治疗乙肝病毒(HBV)携带的肺结核患者临床疗效及安全性.方法:97例HBV携带的肺结核患者,随机分为治疗组50例予以3HREV/6HR治疗,对照组47例予以3HRZE/6HR治疗.结果:治疗组肝功能损害几率24.0%,对照组肝功能损害几率48.9%,两组差异有显著性(P<0.05).治疗组抗结核总有效率92.0%,对照组总有效率93.6%,抗结核化疗总有效率无显著性差异(P>0.05).结论:左氧氟沙星治疗HBV携带的肺结核患者疗效好,肝毒性小,值得临床推广.     

一期前路病灶清除、后路内固定并横突间植骨融合治疗胸腰椎结核

目的:评价一期前路病灶清除、后路内固定并横突间植骨融合治疗胸腰椎结核的临床疗效。方法:采用一期前路病灶清除、后路内固定并横突间植骨融合手术治疗胸、腰椎结核患者17例,按照Frankel分级评定患者手术前后的神经功能,根据X线片评价植骨融合时间,测量术前、术后后凸角度及随访期内的角度丢失。结果:17例患者结核病灶清除彻底,切口均Ⅰ期愈合、无窦道形成,结核治愈无复发。后凸畸形平均矫正19.2°;在随访期内,后凸畸形矫正有1°~4°丢失。X线片示植骨界面骨性融合时间平均5个月。3例出现并发症,对症处理后好转。结论:一期前路病灶清除、后路内固定并横突间植骨融合治疗胸、腰椎结核的疗效确切,具有迅速缓解症状、早期离床活动和较理想的脊柱矫形等优点,是治疗胸、腰椎结核的有效方法。     

影像设备引导下经皮穿刺与支气管填塞治疗耐多药肺结核空洞的疗效分析

目的评价影像设备引导下,经皮穿刺与经支气管填塞,治疗耐多药肺结核空洞的临床疗效。方法CR、CT及生化,病理学确诊的耐多药肺结核空洞患者88例,用经皮穿刺治疗44例(A组)和经支气管填塞治疗44例(B组)。并对照分析其方法,并发症和疗效,评估其应用价值。结果A组闭塞性空洞愈合14例(32%)、净化性空洞愈合16例(36%),无变化及轻微缩小14例(32%),总治愈率为68%;B组闭塞性空洞愈合23例(52.27%),净化空洞愈合17例(38.64%),无变化及轻微变小4例(9%),总治愈率达91%。B组比A组总疗效高出23%(91%~68%),并发症低,疗程缩短1~2个月。结论支气管填塞治疗耐多药肺结核空洞的疗效要优于经皮穿刺的方法。是目前治疗耐多药肺结核空洞的比较有效方法。     

Comparison of redox and D29 phage methods for detection of isoniazid and rifampicin resistance in Mycobacterium tuberculosis

Rapid, accurate and inexpensive methods are essential to detect drug-resistant Mycobacterium tuberculosis and allow timely application of effective treatment and precautions to prevent transmission. The proportion method, the MTT and Alamar Blue redox methods, and the D29 mycobacteriophage assay, were compared for their ability to detect resistance to isoniazid and rifampicin. When tested against a panel of known M. tuberculosis strains, the redox methods and the D29 assay showed good sensitivity and specificity compared to the proportion method, suggesting that they could be useful alternatives for identifying multidrug resistance in M. tuberculosis.     

Mycobacterium Tuberculosis Infection Incidence in Hospitalized Renal Transplant Patients in the United States, 1998–2000

The incidence, risk factors, and prognosis for Mycobacterium tuberculosis (MTB) infection have not been reported in a national population of renal transplant recipients. We performed a retrospective cohort study of 15,870 Medicare patients who received renal transplants from January 1, 1998 to July 31, 2000. Cox regression analysis derived adjusted hazard ratios (AHR) for factors associated with a diagnosis of MTB infection (by Medicare Institutional Claims) and the association of MTB infection with survival. There were 66 renal transplant recipients diagnosed with tuberculosis infection after transplant (2.5 cases per 1000 person years at risk, with some falling off of cases over time). The most common diagnosis was pulmonary TB (41 cases). In Cox regression analysis, only systemic lupus erythematosus (SLE) was independently associated with TB. Mortality after TB was diagnosed was 23% at 1 year, which was significantly higher than in renal transplant recipients without TB (AHR, 4.13, 95% CI, 2.21, 7.71, p < 0.001). Although uncommon, MTB infection is associated with a substantially increased risk of mortality after renal transplantation. High-risk groups, particularly those with SLE prior to transplant, might benefit from intensified screening.     

PCR-SSCP法检测结核分枝杆菌耐药性

目的：探讨耐多药结核分枝杆菌耐药基因突变与耐药性的关系以及聚合酶链反应-单链构象多态性分析(poly merase chain reaction-single strand cinfomlation polymorphism，PCR-SSCP)方法的临床应用价值。方法：用PCR-SSCP方法检测58株结核分枝杆菌临床分离株katG，rpoB，rpsL基因突变并与常规药敏试验检测结果进行对比。结果：经常规药敏试验检测，58株结核分枝杆菌临床分离株中共有41株耐药，其中，耐异烟肼(INH)为35株，高耐药株27株；耐利福平(RFP)为31株，高耐药株24株；耐链霉素(SM)有31株，高耐药株26株。同时耐3种药物的有21株(51．2％)，耐2种药物的14株(34．1％)，单耐药株6株(14．6％)．PCR-SSCP方法对58株临床分离株katG，rpoB，rpsL基因突变的检测率为40％(23／58)，45％(26／58)，38％(22／58)，其中检出3个基因同时突变的有13株(32％)，2种基因突变的12株(29％)，1种基因突变的有10株(23．8％)．常规药敏试验与PCR-SSCP法检出结核分枝杆菌同时耐3种药物的符合率为61．9％(13／21)，检出耐2种药物的符合率为85．70k，(12／14)，检出耐1种药物的符合率为50％(3／6)．高耐药株中突变率为80．5％(62／77)，低耐药株中突变率为60％(12／20)．结论：PCR-SS-CP方法对耐2种以上药物的结核杆菌检出率较高，且耐药基因突变率随着耐药浓度增高而增高。将PCR-SSCP法与药敏试验联合应用可互相弥补，已成为临床指导用药的好方法。     

病灶清除一期植骨内固定治疗脊柱结核

目的探讨脊柱结核病灶清除手术后内固定方式的选择。方法对35例脊柱结核患者采用3种不同术式：16例前路病灶清除、植骨,一期前路钢板或钉棒内固定;14例前路病灶清除、植骨,一期后路经弓根钉棒内固定;5例前路病灶清除、植骨,一期前路钢板、后路钉棒联合内固定。结果35例均获随访,时间18-24个月。X线显示病灶静止,植骨融合,内固定无松动断裂,后凸畸型改善,其中合并脊髓损害15例,按Frankel分级,C级3例、D级12例术后全部恢复到E级。结论病灶清除一期植骨内固定有利于重建脊柱稳定性,纠正和预防脊柱后凸畸形,可避免或减少结核复发。