Monitoring intravesical therapy for superficial bladder cancer using fluorescence in situ hybridization

PURPOSE: We evaluated fluorescence in situ hybridization (FISH) for assessing the response to therapy in patients with superficial bladder cancer receiving bacillus Calmette-Guerin or other intravesical therapies. MATERIALS AND METHODS: A total of 37 patients receiving intravesical therapy for superficial bladder cancer were enrolled in this study. Urine specimens were collected for FISH analysis just prior to the first intravesical therapy in 31 cases and just prior to or within 2 months following the last intravesical therapy in 37. FISH was done using the UroVysion probe set (Abbott Laboratories, Abbott Park, Illinois) with results considered positive if 5 or more cells demonstrated polysomy. Biopsy, cystoscopy and/or cytology results were then compared to FISH results to evaluate the usefulness of the test for monitoring intravesical therapy. RESULTS: Of the patients 25 had a negative and 12 had a positive post-therapy FISH result. All 12 patients with a positive post-therapy FISH result had tumor recurrence, while tumor recurrence was observed in 13 of the 25 with a negative post-therapy FISH result (HR 4.6, 95% CI 1.9 to 11.1, p <0.001). Of the patients with tumor recurrence 7 of 12 with a positive post-therapy FISH result had muscle invasive tumor and 2 of 25 with a negative post-therapy FISH result had muscle invasive tumor (HR 9.4, 95% CI 1.9 to 45.3, p = 0.001). CONCLUSIONS: FISH appears to be useful for monitoring patients with superficial bladder cancer for the response to intravesical therapy. Patients with a positive FISH result at the end of treatment are at high risk for progression to muscle invasive bladder cancer.     

结核分支杆菌14KDa蛋白的基因克隆、表达、纯化及其抗原性的研究

目的基因克隆、表达、纯化结核分枝杆菌14KDa蛋白,研究其抗原性,评价其在血清学诊断中的价值。方法以结核分枝杆菌H37Rv基因组DNA为模板,应用PCR技术扩增14KDa蛋白基因片断,将其插入到高效表达载体PET-22b（＋）上,构建重组质粒PET-22b（＋）/14KDa,重组质粒在大肠杆菌中由IPTG诱导表达,表达产物经金属离子鳌合亲和层析方法纯化,免疫印迹和酶联免疫吸附（ELISA）试验分析重组蛋白的抗原性。结果构建了具有正确基因序列的14KDa蛋白重组质粒,在大肠杆菌BL21（DE3）中以可溶性蛋白形式表达,重组蛋白的表达量占菌体蛋白的40.8%,经过一步金属离子鳌合亲和层析后得到纯度为94.3%的目的蛋白。免疫印迹试验结果表明该蛋白能与羊抗结核血清发生特异免疫结合反应。应用ELISA方法对结核血清参考品进行检测,敏感性和特异性分别为75.6%和96.0%。结论获得了能高效表达14KDa蛋白的大肠杆菌工程菌,目的蛋白以可溶性形式表达,该重组蛋白具有良好的免疫原性,可望成为结核血清学的诊断抗原之一。     

BACTEC MGIT^TM960全自动分枝杆菌培养鉴定／药敏仪基本原理及其应用

BACTEC MGIT^TM960全自动分枝杆菌培养鉴定／药敏仪，能大大缩短分枝杆菌的检测时间，该仪器自动化程度高，无放射性污染，可进行药敏学动态研究。     

Generation of self‐renewing immature dendritic cells from mouse spleen that can take up mycobacteria and present antigens to T cells

Pal R, Marwaha S, Pepponi I, Mann JFS, Paul MJ, Reljic R. Generation of self‐renewing immature dendritic cells from mouse spleen that can take up mycobacteria and present antigens to T cells. APMIS 2010; 118: 729–38. Dendritic cells (DC) play a key role in driving the adaptive immune response to Mycobacterium tuberculosis (MTB), the causative pathogen of tuberculosis (TB). However, studying these important yet very sparse immune cells in the context of MTB pathogenesis is severely restricted by the lack of suitable cell lines and the complexity of culturing of DC progenitors, usually obtained from the bone marrow. However, significant advances have been made towards generating long‐term DC cultures from various lymphoid tissues. Here, we report the evidence for generating a long‐term, self‐renewing DC culture from the Balb/c mouse spleen. We demonstrate that these cells, termed IDC‐3, have a myeloid DC origin, i.e. they are CD11c⁺CD11b⁺⁺CD8‐α^?F4/80^+/? and that they also display a phenotype MHC‐II⁺CD16/32⁺⁺CD80^+/?CD86⁺, indicating that they are immature DC. Following incubation with Mycobacterium bovis BCG (Bacillus Calmette Guerin), the IDC‐3 efficiently took up bacteria and acquired the morphology of mature DC. Importantly though, when IDC‐3 were pre‐stimulated with a mycobacterial antigen in vitro, they were able to induce proliferation of T lymphocytes from mice immunized with the same antigen. The T‐cell stimulatory potential of IDC‐3 was further enhanced when the cells were co‐stimulated with an anti‐CD40 mAb. We therefore suggest that the IDC‐3 culture system could be a useful tool for studying the interaction of DC with mycobacteria.     

肺结核患者初始耐药的流行病学分析

目的了解安徽省结核分支杆菌初始耐药的流行病学特征.方法将安徽省肺科医院1997年初～1999年底3年间230例初治肺结核患者的初始耐药性进行流行病学分析.结果3年的初始耐药率分别为:1997年30.4%,1998年27.1%,1999年26.3%,呈逐年下降趋势;异烟肼(INH)、链霉素(SM)、利福平(RFP)的初始耐药率也呈逐年下降趋势.城市初始耐药率(29.7%)高于农村(28.3%)和城镇(21.6%);中老年组初始耐药率高于青年、儿童组;不同职业中,知识分子具有较高的初始耐药率33.3%,学生次之29%,农民为27.7%,工人为25%;结论本地区结核病控制工作取得了一定的成效,但初始耐药情况仍十分严重.     

蛋白芯片法与金标法及抗酸染色法诊断结核病的比较

目的将诊断结核病的蛋白芯片法与金标法及抗酸染色法进行比较,以评价蛋白芯片的临床应用价值。方法用蛋白芯片法、金标法及抗酸染色法检测75例确诊结核病人、30例肺部其它疾病患者和健康体检者的血清标本,并进行相关统计分析。结果蛋白芯片法、金标法及抗酸染色法检出结核分枝杆菌的灵敏度为64%,72%及22.7%,前两者比较无显著性差异(P>0.05),但均高于抗酸染色法(P<0.05)。三者的特异性分别为90%,76.7%及96.7%。结论蛋白芯片法检测结核分枝杆菌,与金标法及抗酸染色法比较,具有简便、快速、敏感性较高和特异性强等优点,是临床辅助诊断结核病的有效方法。     

江苏省扬州地区41例非结核分歧杆菌肺病的流行病学及临床特征分析

目的 探讨扬州地区非结核分歧杆菌肺病的流行病学特点及临床特征.方法 对本院收治的41例非结核分歧杆菌肺病的临床资料进行回顾分析.结果 治疗6月末,34例患者临床症状缓解,21例患者痰菌阴转.结论 扬州地区非结核分歧杆菌肺病多见于中老年及男性患者,农村居民多于城市居民,常合并有基础疾病,误诊率高,未见人传人现象.肺内病灶较广泛,多有空洞,常为薄壁空洞,耐药率高.经积极治疗后临床症状可缓解,肺内病灶可吸收好转,但痰菌阴转率低.