脂多糖对间充质干细胞Toll样受体4基因的表达水平及活性的影响

目的 探讨脂多糖(LPS)对骨髓间充质干细胞(MSC)Toll样受体4(TLR-4)基因的表达及其功能的影响.方法 采用贴壁培养和密度梯度离心法从大鼠骨髓分离MSC,通过细胞形态、成骨分化潜能及流式细胞术检测表型以鉴定其纯度;半定量RT-PCR和流式细胞术分别检测MSC在不同浓度(1、10、100 μg/ml)LPS存在条件下培养24 h的TLR-4 mRNA相对表达量和共刺激分子(CD80、CD86、MHC-Ⅱ)的表达水平;ELISA法定量检测TNF-α的分泌水平.结果 骨髓MSC低表达TLR-4mRNA(相对表达量0.61±0.10),同时表达CD80[(9.56±0.69)%]、CD86[(22.03±2.03)%]、MHC-Ⅱ[(2.51±0.97)%],少量分泌TNF-α[(4.97±2.98)pg/ml].MSC经LPS处理后,其TLR-4mRNA、共刺激分子表达和TNF-oα分泌水平均升高,其中10 μg/ml LPS培养组升高显著,TLR-4 mRNA相对表达水平为1.55±0.02;CD80、CD86、MHC-Ⅱ阳性细胞率分别为(41.70±2.92)%、(59.72±2.00)%、(24.56±2.19)%;TNF-α分泌水平为(213.12±69.08)pg/ml,与对照组比较,P值均＜0.01.100μg/ml LPS处理组与10 μg/mlLPS处理组相比,各项检测指标均降低,但MHC-Ⅱ和TNF-α的降低水平无统计学意义(P＞0.05).结论 骨髓MSC低表达TLR-4,体外LPS可促进骨髓MSCTLR-4的表达,且与浓度相关.伴随TLR-4表达水平的升高,CD80、CD86、MHC-Ⅱ表达水平及TNF-α水平同时升高.     

精浆肉碱测定在男性生育力评估和弱精子症疗效监测中的应用*

摘要:目的比较正常 男性与弱精子症男性精浆中肉碱含量,评估精浆肉碱水平与前向运动精子百分率( PR)之间的一致性,并外源性补充肉碱对弱精子症患者的影响。方法收集511 例正常可育男性及弱精子症患者精液样本,采用精浆肉碱检测试 剂盒(固定时间法)测定精浆肉碱,比较两组水平差异及与PR的一致性。另选77例弱精子症患者给予左卡尼汀(1 g/次,3次/日,30日/疗程)治疗,监测服药前后精浆肉碱及PR变化。结果弱精子 症患者组精浆肉碱含量[ ( 194.34+65.41) μmol/L] 显著低于正常可育男性组[ ( 405.43士72.12) μumol/L] (P<0.01);以精浆肉碱≥325 μmol/L为阈值, Kappa值为0.81,诊断符合率达93.74% ;给予左卡尼汀治疗后,弱精子症组精浆肉碱浓度[ (356.03+84.87) μumol/L]较之前[(183.61+ 79.54) μumol/L]明显上升,PR[ ( 32.69+8.35)%]较之前[ (16.56+7.74)%]显著升高(P均<0.01)。结论精浆 肉碱检测试剂盒能准确高效检测大量临床样本,可用于弱精子症诊断及疗效评估。外源性补充肉碱可提高弱精子症患者精浆肉碱水平和精子活力,有助于改善其生育能力。     

保和丸治疗小儿口疮

保和丸出自于《丹溪心法》,为治疗食积证的代表方剂,临床用之甚广。笔者在辨证论治思想的指导下用其治疗小儿口疮,每获良效。     

精液常规参数初检合格后不同时间点再分析结果及其与精子DNA损伤的关系

目的:探讨完全液化且常规参数初检合格的精液标本,于不同时间再分析的结果差异,及精子DNA碎片化指数(DFI)与精子活动力改变的相关性。方法:选取127份符合纳入标准的精液标本,分别于取样后15、30、60 min时采用计算机辅助精液分析(CASA)系统进行分析。精子形态分析采用Shorr染色法,吖啶橙试验(AOT)检测DFI。结果:3个时间点精子浓度、a级和b级精子百分率均无统计学差异(P>0.05)。取样15 min时a+b和a+b+c级精子百分率显著高于30和60 min时的结果(P<0.05),后两者间无统计学差异(P>0.05)。不同时间精子活动力各项指标中,至少有1项由"正常组"变为"异常组"的发生率为25.2%,两组间DFI和形态学无统计学差异(P>0.05)。取样后15到60 min变化的精子活动力指标中,a、a+b、a+b+c级下降值与DFI存在正相关性(P<0.05)。结论:取样后15 min内完全液化并初查参数合格的精液标本,30～60 min内复查时,a级和b级精子百分率波动并无显著差异,而a+b级及a+b+c级精子则可能显著下降,精子活动力指标可能出现异常。故应至少进行2次精液分析,综合评估生育力。如2次结果差异较大,尤其是a级精子百分率下降幅度较大,则可能与精子DNA损伤有关,需进一步行精子DNA损伤检测。     

射频消融治疗甲状腺乳头状癌的研究进展

射频消融（RFA）是在超声引导下利用高频交变电流所释放出的能量使局部组织产生高温,使病灶凝固坏死,进而消除病灶的一种热消融方法。RFA技术现已广泛应用于肝癌、肾癌、肺癌和甲状腺良性结节等疾病的治疗中。对于甲状腺乳头状癌（PTC）最主要的治疗方式仍是手术治疗,然而手术治疗可能会带来各种并发症,增加患者的心理负担。目前RFA已经应用于PTC原发灶、PTC局部复发灶、PTC转移淋巴结及PTC远处转移的治疗当中。另外,在RFA治疗甲状腺疾病过程中,必须严格把握其适应证。本文通过阐述RFA、RFA在PTC中的应用及RFA的并发症,对RFA技术应用于PTC的研究进展作一综述。     

恒河猴P21基因在COS-7细胞中沉默位点的验证

【摘要】 目的 在细胞水平筛选恒河猴 p21基因的有效沉默靶点。方法 检测 COS-7 中 p21基因的表达水平;设计shRNA并构建p21-RNA干扰慢病毒载体FUGW-TDT-p21shRNA转染 COS-7 细胞,通过real-time PCR检测沉默效率,并以 Western Blot 在蛋白水平进行验证。结果 筛选到四个有效的靶位,分别位于p21 mRNA 的541-561bp,542-562bp,215-239bp,624-648bp。四个靶位点在mRNA水平的沉默效率分别为 (91.82?.21)%, (82.47?.48)%, (81.31?.69)%和(87.35?.59)%。相应的蛋白表达量为(11.97?.70)%, (20.22?.65)%, (23.21?.63)%和(14.42?.86)%。结论 在细胞水平筛选得到四个有效的 p21基因沉默靶点,可用于恒河猴基因沉默研究。     

抗原提呈细胞与同种异基因移植后免疫重建

在骨髓移植和造血干/祖细胞移植等造血细胞移植后，机会感染的易感性是随着供者来源的免疫系统恢复时间的延长而增加的，因此，移植后宿主的免疫系统的及时重建对于移植物的最终植活和抗移植后并发症等都有着重要的意义。研究显示：提高输注的造血干／祖细胞的剂量，可使造血系统植活和免疫系统重建的时间明显缩短，重建的免疫细胞不仅在“量”上能满足宿主的需要，而且，在其“质”上[例如：T细胞的白细胞介素-2（IL-2）和干扰素-γ（INF-γ）的产量等]也是有保障的；并且，将骨髓等移植物中的T细胞去除而避免急性移植物抗宿主病（aGVHD）的发生，也是这种免疫重建加速趋势形成的主要原因。     

短QT间期发生室性心律失常的电生理机制探讨

目的了解短QT间期发生室性心律失常的电生理机制。方法应用吡那地尔在家兔左室楔形灌注组织建立短QT模型,利用标准玻璃微电极技术记录心外膜下、心内膜下及中层心肌细胞动作电位,并观测三层心肌细胞复极达90%的动作电位(APD90)及跨壁复极离散度(TDR)在吡那地尔、吡那地尔+异丙肾上腺素、奎尼丁、glybenclamide作用下的变化。采用S1S2程序刺激,观测在各种条件下心律失常的诱发状况。结果吡那地尔明显缩短APD90且伴有TDR增大(58.84±13.42ms vs35.26±13.30ms),并可诱发出异常心肌搏动。异丙肾上腺素可增大吡那地尔的该作用(64.60±21.46ms vs58.84±13.42ms),而奎尼丁和glybenclamide则可逆转吡那地尔的此作用,并减少异常搏动的发生。结论TDR增大可能是短QT综合征易于发生致命性心律失常的基础,而奎尼丁通过减小室壁心肌细胞的不均一性而对短QT综合征起到治疗作用。     

食管造影及温度监测指导环肺静脉消融治疗心房颤动

Objective Because of the severe consequences of an atrial esophageal fistula,it is vital to avoid this complication. The most safe way is to avoid ablating the segment of esophagus behind left atrium (LA). What we do is to image the relationship of esophagus to posterior LA wall and real-time esophageal tem-perature monitoring to ablate the posterior LA wall. Methods Sixty-four patients with paroxysmal atrial fibrilla-tion (PAF) were enrolled to pulmonary veins (PV) isolation. Swallowing a radiocontrast agent at the same time when imaging of LA to observe the relationship of esophagus to posterior LA wall and according to the different relationship between the segment of esophagus behind LA and PV, the esophagus were divided into type Ⅰ , type Ⅱ and type Ⅲ. A esophageal temperature probe was inserted and advanced into the esophagus directly posterior to the LA. Three steps ablation strategy and real-time esophageal temperature monitoring were applied to guide the PV isolation. The procedure was interrupted when the esophageal temperature was 39℃ until] the tempera-ture renormalized and ended when PV were isolated. Results There were 48 type ⅠI , 11 type Ⅱ and 5 type Ⅲ esophagus in the 64 patients. After three steps ablation,all PV isolations were completed. Only 18.8% of the patients needed to ablate the posterior LA close to the esophagus. Conclusion Only one PV ostiolum close to the posterior LA wall in most PAF patients and many of them can achieve complete PV isolation without ablating the posterior LA close to esophagus which could minimize the risk of esophageal injury dramatically.     

食管造影及温度监测指导环肺静脉消融治疗心房颤动

Objective Because of the severe consequences of an atrial esophageal fistula,it is vital to avoid this complication. The most safe way is to avoid ablating the segment of esophagus behind left atrium (LA). What we do is to image the relationship of esophagus to posterior LA wall and real-time esophageal tem-perature monitoring to ablate the posterior LA wall. Methods Sixty-four patients with paroxysmal atrial fibrilla-tion (PAF) were enrolled to pulmonary veins (PV) isolation. Swallowing a radiocontrast agent at the same time when imaging of LA to observe the relationship of esophagus to posterior LA wall and according to the different relationship between the segment of esophagus behind LA and PV, the esophagus were divided into type Ⅰ , type Ⅱ and type Ⅲ. A esophageal temperature probe was inserted and advanced into the esophagus directly posterior to the LA. Three steps ablation strategy and real-time esophageal temperature monitoring were applied to guide the PV isolation. The procedure was interrupted when the esophageal temperature was 39℃ until] the tempera-ture renormalized and ended when PV were isolated. Results There were 48 type ⅠI , 11 type Ⅱ and 5 type Ⅲ esophagus in the 64 patients. After three steps ablation,all PV isolations were completed. Only 18.8% of the patients needed to ablate the posterior LA close to the esophagus. Conclusion Only one PV ostiolum close to the posterior LA wall in most PAF patients and many of them can achieve complete PV isolation without ablating the posterior LA close to esophagus which could minimize the risk of esophageal injury dramatically.