胸苷激酶基因系统对膀胱癌细胞及凋亡基因的影响

目的 探讨单纯疱疹病毒胸苷激酶／丙氧鸟苷(HSV-TK／GCV)系统对膀胱癌细胞BIU87的杀伤作用及对凋亡基因的影响。方法 脂质体将TK基因成功转入BIU87。噻唑蓝(MTT)法、TUNEL法、流式细胞仪(FCM)分别检测GCV对其生长影响及凋亡率和细胞周期变化；逆转录一聚合酶链反应(RT-PCR)检测bcl-2、bax、bak、Caspase-3 mRNA表达变化．比色法测定Caspase．3活性变化。结果 作用5dlmg,／LGCV能杀死67％的细胞，100mg／，LGCV达到最大杀伤率杀死97％的细胞；GCV诱导细胞凋亡，细胞周期阻滞在S和G2期；bcl-2表达降低，bax表达升高，Caspase-3表达及活性升高。结论 HSV-TK／GCV系统可有效杀伤膀胱癌细胞并引起细胞凋亡，细胞周期阻滞、bcl-2和bax表达变化、Caspase-3活化是引起凋亡发生的重要因素。     

含精氨酸-甘氨酸-天冬氨酸肽白细胞介素-24突变体蛋白对肝癌细胞抑制作用

目的 观察含精氨酸-甘氨酸-天冬氨酸(RGD)肽模体的白细胞介素(IL)-24突变体蛋白(RGD-IL-24)对肝癌细胞的抑制作用.方法 肝癌HepG2细胞分别加入各10μl的RGD-IL-24(8 mg/L)、IL-24(8 mg/L)和磷酸盐缓冲液(PBS),噻唑蓝(MTT)比色法检测HepG2细胞生长抑制作用;DAPI染色检测HepG2细胞凋亡;免疫印迹法检测HepG2细胞bax、bcl-2及Caspase-3蛋白表达;黏附实验检测与HepG2细胞的靶向黏附.结果 RGD-IL-24、IL-24治疗4 d后HepG2细胞存活率分别为(0.219±0.015)、(0.397±0.009),与对照组(0.823±0.013)比较差异有统计学意义(P<0.01).RGD-IL-24、IL-24治疗组细胞凋亡率分别为(0.631±0.027)、(0.472±0.031),与对照组(0.082±0.013)比较差异有统计学意义(P<0.01).RGD-IL-24抑制HepG2细胞生长、诱导凋亡效果均比IL-24显著增强(P<0.05).与IL-24治疗组比较,RGD-IL-24治疗组HepG2细胞促凋亡蛋白bax增加、抗凋亡蛋白bcl-2减少、活化Caspase-3蛋白量增加.黏附实验证实RGD-IL-24与HepG2细胞的靶向结合作用增强.结论 含RGD肽模体的IL-24蛋白能通过与肝癌HepG2细胞的靶向结合增强其凋亡诱导作用.     

绿脓杆菌菌毛株对肝癌细胞Hep-2生长的抑制作用

目的 观察绿脓杆菌菌毛株菌苗(PA-MSHA)对人肝细胞癌细胞株HepG-2的抑制作用及机制.方法 噻唑蓝(MTT)法检测细胞生长抑制情况.原位末端标记(TUNEL)法检测细胞凋亡情况;透射电镜观察细胞超微结构.Western blot法检测bcl-2、bax、Caspase-3蛋白表达;裸鼠移植瘤内及瘤周药物注射观察不同药物浓度及作用时间的体内抑瘤作用.结果 肿瘤细胞的抑制率与药物浓度和作用时间成正比;高、低浓度实验组和对照组凋亡率差异有统计学意义(P＜0.01);电镜观察可见经PA-MSHA作用后的HepG2细胞呈现典型的凋亡表现;PA-MSHA作用后可引起bax、Caspase-3蛋白表达上调,bcl-2蛋白表达下调;高浓度PA-MSHA可明显抑制裸鼠移植瘤的生长,17 d抑瘤率为60%.结论 PA-MSHA可明显抑制HepG-2细胞的生长,诱导凋亡是其另外一个作用机制.     

NDGA对HepG2细胞5-LOX及其凋亡相关基因表达的影响

目的 探讨脂氧合酶(lipoxygenase)抑制剂NDGA对肝癌细胞HepG2细胞5-LOX及其凋亡相关基因hTERT、bcl-2及bax表达的影响.方法 用RT-PCR方法检测NDGA对HepG2细胞5-LOX及其凋亡相关基因hTERT、bcl-2及bax表达的影响.结果 25、50、100、200 μmol/L的NDGA作用HepG2细胞24、48 h后5-LOX表达逐渐减弱,hTERTmRNA、bcl-2的表达明显下调,而bax的表达逐渐上调.与未加NDGA干预的HepG2对照组比较,P＜0.05.结论 5-LOX在HepG2细胞中存在高表达,提示5-LOX的高表达在肝细胞癌发病过程中可能起着重要作用;而端粒酶hTERTmRNA、bcl-2及bax等凋亡相关基因可能参与了其发病过程.脂氧合酶抑制剂NDGA体外可有效抑制细胞5-LOX的表达,明显下调端粒酶hTERTmRNA、bcl-2及上调bax的表达.LOX可能是肝癌生物化学治疗的新靶点.     

新辅助化疗对结肠癌细胞凋亡及相关调控蛋白表达的影响

目的 观察新辅助化疗(NACT)对结肠癌细胞凋亡及相关调控蛋白表达的影响.方法 收集结肠癌患者114例,采用原位末端标记法和免疫组织化学方法分别观察经NACT的FOLFOX4方案治疗前后结肠癌细胞凋亡和癌组织中B细胞淋巴瘤/白血病-2相关X蛋白(bax)、B细胞淋巴瘤/白血病-2(bcl-2)、环氧合酶-2(COX-2)、p53蛋白的表达,并判定NACT的近期疗效.结果 NACT化疗后结肠癌细胞发生典型凋亡形态学变化,细胞凋亡系数(3.87％比1.17％)和凋亡阻性率(59.65％比23.68％)显著高于化疗前.经NACT治疗后患者结肠癌细胞bcl-2(54.39％比70.15％)、COX-2(28.07％比50.00％)和p53(45.61％比70.05％)的阳性表达率与化疗前比较均有不同程度降低,差异有统计学意义(P＜0.05或P＜0.01),而bax阳性表达率(22.81％比50.88％)显著增强,差异有统计学意义(P＜0.01).同时bcl-2/bax比值(3.08比1.07)也显著降低.NACT治疗组的总有效率(39.47％)显著高于对照组(1.75％),治疗后患者无严重药物不良反应.结论 结肠癌患者经NACT化疗后癌组织中bcl-2、COX-2和p53蛋白表达明显降低,而bax蛋白的表达显著增加,促进癌细胞凋亡,且NACT的近期疗效显著.     

胃癌凋亡相关蛋白Survivin、B淋巴细胞/白血病-2、bax表达与肿瘤细胞体外化疗药敏性的关系

目的 探讨胃癌凋亡相关蛋白表达与体外化疗药敏性的关系. 方法 对64例胃癌标本进行Survivin、R淋巴细胞/白血病-2(bcl-2)、bax免疫组织化学染色,并以噻唑蓝(MTT)比色法检测体外化疗药物敏感性.结果 肿瘤Survivin、bcl-2、bax阳性表达率分别为90.6％、75.0％、68.8％;Survivin与bax、bcl-2与bax 间表达强度均呈负相关(r=-0.45044、-0.414 03,P＜0.01).在耐药因子表达程度与药物对肿瘤细胞抑制率的关系中,Survivin强表达时,表阿霉素(eADM)、顺铂(DDP)对肿瘤细胞的抑制率明显降低(P＜0.05),但奥沙利铂(L-OHP)对肿瘤细胞的抑制率明显增加(P＜0.05);bcl-2强表达时,5-氟尿嘧啶(5-Fu)、紫杉醇(PTX)、eADM对肿瘤细胞的抑制率明显低于弱表达组(P＜0.05);bax强表达组中,5-Fu、eADM、L-OHP和甲氨喋呤(MTX)对肿瘤细胞的抑制率明显高于弱表达组(P＜0.05).结论 胃癌凋亡相关蛋白表达与部分化疗药物敏感性有关.     

17β雌二醇、他莫昔芬和雷洛昔芬诱导前列腺癌细胞凋亡及细胞周期阻滞的研究

目的探讨17β雌二醇(E2)、他莫昔芬(TAM)和雷洛昔芬(RAL)对前列腺癌PC3凋亡、细胞周期的影响及作用机制。方法检测不同浓度E2、TAM、RAL作用于PC3细胞不同时间后细胞生长抑制率和48 h后细胞周期分布、凋亡率、bcl-2和Caspase-3蛋白的表达,以及12 h后p21WAF1 mRNA．表达。Hoechst染色和电镜观察细胞凋亡。结果E2、TAM、RAL呈时间及浓度依赖性抑制PC3细胞增殖。10-4 mol／L E2、10-5 mol／L TAM和10-5 mol／L RAL作用48 h后检测到凋亡,凋亡率分别为(21．54±0．91)％、(38．28±1．16)％、(42．41±2．26)％(P<0．05),bcl-2和Caspase-3分别为对照组的0．5、0．4、0．35倍;1．4、1．6、1．6倍。细胞阻滞在G1期。对照组和处理组p21WAF1基因表达强度分别为1．12、3．31、5．24、4．48。结论E2、TAM、RAL可以增强p21WAF1基因表达使PC3阻滞于G1期,通过下调bcl-2蛋白并上调Caspase-3蛋白诱导PC3凋亡。     

Ad-ING4-IL-24双基因共表达腺病毒对前列腺癌细胞株PC3体内外抑癌增效作用

目的 观察肿瘤生长抑制因子4(ING4)和白细胞介素-24(IL-24)双基因共表达腺病毒载体对PC3人前列腺癌细胞体内外抑癌增效作用.方法 应用半定量逆转录-聚合酶链反应(RT-PCR)和Western blot法检测ING4和IL-24在PC3细胞中的表达;噻唑蓝(MTT)比色法和流式细胞术(FCM)检测双基因对PC3细胞的生长抑制和凋亡效应;半定量RT-PCR法和Western blot法检测双基因的表达对PC3细胞中的bcl-2、bax、p53和Caspase-3凋亡相关基因表达的影响.在前列腺癌的裸鼠移植瘤动物模型上,检测Ad-ING4-IL-24对移植瘤生长的抑制作用,并通过免疫组织化学法检测bcl-2、bax、Caspase-3、CD34等相关因子的表达.结果腺病毒介导的ING4和IL-24双基因在PC3细胞中成功表达,对PC3细胞增殖具有抑癌增效功能,可引起p53、bax、Caspase-3基因表达上调和bcl-2基因表达下凋,诱导细胞凋亡.Ad-ING4-IL-24能显著抑制PC3裸鼠前列腺癌移植瘤生长,瘤重的抑制率可达74%,免疫组织化学结果显示Ad-ING4-IL-24能明显上调bax和Caspase-3的表达和下调bcl-2和CD34基因表达.结论 Ad-ING4-IL-24在体内外均可明显抑制PC3细胞的生长,诱导其凋亡,具有抑癌增效功能.其机制可能是通过上凋p53、bax、Caspase-3基因和下凋bcl-2和CD34基因表达水平.     

力学刺激对软骨细胞凋亡信号转导分子Caspase-3及bcl-2、bax mRNA表达的影响

目的 观察力学刺激对软骨细胞凋亡信号转导分子半胱氨酸酶-3( Caspase-3)及B细胞淋巴瘤-2(bcl-2)、bax mRNA表达和凋亡的影响.方法 兔膝关节软骨分离培养,在第3代软骨细胞培养瓶中加入不同剂量的Caspase-3、bcl-2、bax抑制剂,力学刺激诱导凋亡,然后检测软骨细胞凋亡率,聚合酶链反应(PCR)半定量分析Caspase-3 bcl-2、bax mRNA表达.结果 力学刺激诱导软骨细胞凋亡,在加入抑制剂的各组和空白组的凋亡率差异有统计学意义(P＜0.05);各组Caspase-3及bcl-2、bax mRNA表达和空白组差异有统计学意义(P＜0.05).Caspase-3抑制剂组的凋亡率和Caspase-3表达明显相关(r=0.69,t=3.41,P＜0.01);bcl-2抑制剂组和bcl-2的表达明显相关(r=0.73,t=3.97,P＜0.01);bax抑制剂组和bax的表达明显相关(r=0.89,t =6.69,P＜0.01);各组差异均有统计学意义.结论 Caspase-3、bax抑制剂能对抗力学刺激诱导的凋亡,而bcl-2抑制剂使凋亡增加,各组Caspase-3及bcl-2、bax mRNA表达发生相应改变.     

FasL、抗DR5单克隆抗体对结肠癌细胞株HT29的杀伤作用及其机制

目的 探讨FasL、抗DR5单克隆抗体(Anti-DR5 mAb)对结肠癌细胞株HT29的杀伤作用及机制.方法 采用逆转录-聚合酶链反应(RT-PCR)、噻唑蓝(MTT)比色法、DNA倍体分析,Western blot等.结果 HT29细胞表面Fas mRNA的表达低于DIL5 mRNA的表达.50 mg/L FasL和Anti-DR5 mAb对HT29细胞的杀伤率分别为(25.49±0.90)%和(48.90±3.15)%,这种作用呈剂量依赖性.流式细胞术分析细胞周期和凋亡实验表明FasL和Anti-DR5 mAb能够抑制HT29细胞的生长,并且诱导它的凋亡.25 mg/L FasL和Anti-DR5 mAb对HT29细胞的凋亡指数分别为(13.8±1.5)%和(22.6±1.1)%.FasL和Anti-DR5 mAb作用HT29细胞后,Caspase-3蛋白表达上升,bcl-2蛋白表达水平下降.结论 FasL、Anti-DR5 mAb能不同程度的诱导结肠癌细胞株HT29凋亡,其机制与其受体和Caspase-3、bcl-2的表达有关.     

Presenteeism and Absenteeism Before and After Total Hip and Knee Arthroplasty

BackgroundAbsenteeism is costly, yet evidence suggests that presenteeism—illness-related reduced productivity at work—is costlier. We quantified employed patients’ presenteeism and absenteeism before and after total joint arthroplasty (TJA).MethodsWe measured presenteeism (0-100 scale, 100 full performance) and absenteeism using the World Health Organization’s Health and Work Performance Questionnaire before and after TJA among a convenience sample of employed patients. We captured detailed information about employment and job characteristics and evaluated how and among whom presenteeism and absenteeism improved.ResultsIn total, 636 primary, unilateral TJA patients responded to an enrollment email, confirmed employment, and completed a preoperative survey (mean age: 62.1 years, 55.3% women). Full at-work performance was reported by 19.7%. Among 520 (81.8%) who responded to a 1-year follow-up, 473 (91.0%) were still employed, and 461 (88.7%) had resumed working. Among patients reporting at baseline and 1 year, average at-work performance improved from 80.7 to 89.4. A Wilcoxon signed-rank test indicated that postoperative performance was significantly higher than preoperative performance (P < .0001). The percentage of patients who reported full at-work performance increased from 20.9% to 36.8% (delta = 15.9%, 95% confidence interval = [10.0%, 21.9%], P < .0001). Presenteeism gains were concentrated among patients who reported declining work performance leading up to surgery. Average changes in absences were relatively small. Combined, the average monthly value lost by employers to presenteeism declined from 15.3% to 8.3% and to absenteeism from 16.9% to 15.5% (ie, mitigated loss of 8.4% of monthly value).ConclusionAmong employed patients before TJA, presenteeism and absenteeism were similarly costly. After, employed patients reported increased performance, concentrated among those with declining performance leading up to surgery.     

The German emergency and disaster medicine and management systemdhistory and present

As well for optimized emergency management in individual cases as for optimized mass medicine in disaster management, the principle of the medical doctors approaching the patient directly and timely, even close to the site of the incident, is a long-standing marker for quality of care and patient survival in Germany. Professional rescue and emergency forces, including medical services, are the “Golden Standard” of emergency management systems. Regulative laws, proper organization of resources, equipment, training and adequate delivery of medical measures are key factors in systematic approaches to manage emergencies and disasters alike and thus save lives. During disasters command, communication, coordination and cooperation are essential to cope with extreme situations, even more so in a globalized world. In this article, we describe the major historical milestones, the current state of the German system in emergency and disaster management and its integration into the broader European approach.     

Differences in Osteocyte Density and Bone Histomorphometry Between Men and Women and Between Healthy and Osteoporotic Subjects

Bone defects related to osteoporosis develop with increasing age and differ between males and females. It is currently thought that the bone remodeling process is supervised by osteocytes in a strain-dependent manner. We have shown an altered response of osteocytes from osteoporotic patients to mechanical loading, and osteocyte density is reduced in osteoporotic patients, which might relate to imperfect bone remodeling, leading to lack of bone mass and strength. Hence, information on osteocyte density will contribute to a better understanding of bone biology in males and females and to the assessment of osteoporosis. Osteocyte density as well as conventional histomorphometric parameters of trabecular bone were determined in cancellous iliac crest bone of healthy postmenopausal women and men and of osteoporotic women and men. Osteocyte density was higher in healthy females than in healthy males and lower in osteoporotic females than in healthy females. Bone mass was reduced in osteoporotic patients, both male and female. In females, trabecular number was reduced, whereas in males, trabecular thickness was reduced and eroded surface was increased. There were no correlations between the parameter groups bone architecture, bone formation, bone resorption, and osteocyte density. These results are consistent with impaired osteoblast function in osteoporotic patients and with a different mechanism of bone loss between men and women, in which osteocyte density might play a role. The reduced osteocyte numbers in female osteoporotic patients might relate to imperfect bone remodeling leading to lack of bone mass and strength. M. G. Mullender and S. D. Tan contributed equally to this work.     

肝内胆管囊腺瘤和囊腺癌的CT、MRI及病理表现

目的探讨肝内胆管囊腺瘤和囊腺癌的CT、MRI和病理特点。方法回顾性分析经手术病理证实的6例肝内胆管囊腺瘤和2例肝内胆管囊腺癌的影像及临床病理资料，将病变的影像表现与其病理大体形态及组织学表现作对照分析。结果6例肝内胆管囊腺瘤，女4例、男2例；2例肝内胆管囊腺癌均为女性病人；8例病人平均年龄55岁。所有病灶均表现为多房囊性肿块，肿瘤囊腔各分房内常为多种液体成分，在CT上可表现为不同密度、在MRI上可表现为不同信号强度。囊内出现多发大小不等的壁结节在胆管囊腺癌内更常见，囊内有分隔但无壁结节只见于胆管囊腺瘤。在7例CT扫描中，4例胆管囊腺瘤和1例胆管囊腺癌可见囊壁或分隔上钙化，囊壁、囊内分隔及囊内结节均为轻、中度延迟增强。肿瘤中出现卵巢样间质见于3例胆管囊腺瘤和1例胆管囊腺癌，且均为女性病人。结论肝内胆管囊腺瘤和囊腺癌是肝脏不常见的囊性肿瘤，影像上多房、囊内有分隔且各分房囊内密度或信号不一致，高度提示肝内胆管囊腺瘤或囊腺癌的诊断，如囊内伴有多发大小不等的结节，则进一步提示囊腺癌的可能。但影像学表现不能区分肿瘤中有无卵巢样间质。