铜绿假单胞菌的药敏与头孢哌酮-舒巴坦等抗菌药物用药频度相关性

目的了解铜绿假单胞菌的药物敏感性(药敏)与头孢哌酮-舒巴坦等抗菌药物用药频度的相关性,为临床合理用药提供依据。方法分别统计我院2002—2008年头孢哌酮-舒巴坦等抗菌药物用药频度和铜绿假单胞菌的药敏,采用SPSS13.0统计软件用Sperman’s相关法对2种变量进行相关性分析。结果抗菌药物中头孢哌酮-舒巴坦耐药率最低,但2008年比2002年中敏率上升了291.0%,敏感率则呈下降趋势。相关性研究结果显示:头孢哌酮-舒巴坦的中敏率与哌拉西林-三唑巴坦、头孢哌酮-舒巴坦、3代头孢类用药频度呈正相关(r=0.857,P<0.05;r=0.929,P<0.01;r=0.786,P<0.05);头孢哌酮-舒巴坦的敏感率与本药、美罗培南、3代头孢类含酶抑制药用药频度呈负相关(r=-0.821,P<0.05;r=-0.786,P<0.05;r=-0.786,P<0.05),与头孢他啶、阿米卡星、左氧氟沙星(0.5g)用药频度呈正相关(r=0.821,P<0.05;r=0.929,P<0.01;r=1.000,P<0.01);左氧氟沙星(0.5g)用药频度分别与环丙沙星、阿米卡星、左氧氟沙星以及头孢哌酮-舒巴坦耐药率呈完全负相关(r=-1.000,P<0.01;r=-1.000,P<0.01;r=-1.000,P<0.01;r=-1.000,P<0.01);美罗培南耐药率与亚胺培南-西司他丁、哌拉西林-三唑巴坦、头孢吡肟耐药率呈完全正相关(r=1.000,P<0.01;r=1.000,P<0.01;r=1.000,P<0.01);头孢他啶和氨曲南的耐药率与其他药物用药频度及耐药率也有相关性。结论头孢哌酮-舒巴坦等抗菌药物的药敏与用药频度存在显著的相关性,近期临床应注意调整抗菌药物用药频度。     

D-氨基葡萄糖衍生物诱导Eca-109细胞凋亡的机制

目的探讨2-(3-羧基-1-丙酰氨基)-2-脱氧-D-葡萄糖{2-[(3-carboxy-1-oxoprogy1)amino]-2-deoxy-D-Glucose,CO-PADG}诱导Eca-109细胞凋亡的机制。方法不同浓度COPADG作用于人食管癌Eca-109细胞24h,检测Eca-109细胞的抑制率、凋亡率、细胞内活性氧(reactive oxygen spe-cies,ROS)、线粒体跨膜电位。结果Eca-109细胞凋亡率与COPADG浓度呈正相关,r=1.0,P<0.01;Eca-109细胞线粒体膜电位水平与Eca-109细胞凋亡率相关,r=1.0,P<0.01;ROS水平与Eca-109细胞凋亡率呈正相关,r=1.0,P<0.01;ROS水平与Eca-109细胞线粒体膜电位水平呈负相关,r=1.0,P<0.01。结论COPADG可促进Eca-109细胞凋亡,提高Eca-109细胞内ROS水平,并降低线粒体膜电位。实验结果提示COPADG提高ROS,降低Eca-109细胞线粒体膜电位启动细胞凋亡通路促使Eca-109细胞凋亡,并且线粒体膜电位的下降是通过提高ROS实现的。     

胡桃醌及其与顺铂联用对宫颈癌HeLa细胞存活和凋亡的影响

目的探讨胡桃醌及其与顺铂联合用药对宫颈癌HeLa细胞存活和凋亡的影响。方法体外培养人宫颈癌HeLa细胞,加入胡桃醌10～200μmo.lL-1或者胡桃醌20μmo.lL-1+顺铂20μmo.lL-1继续培养8 h或24 h,用倒置显微镜观察细胞形态的变化,用MTT法测定细胞存活率,用流式细胞仪检测细胞周期和细胞早期凋亡率。结果胡桃醌10,20,50,100和200μmo.lL-1与HeLa细胞作用24 h,随着胡桃醌浓度的增加,光镜下HeLa细胞逐渐变小、变圆;HeLa细胞存活率明显降低,分别由对照组的(100.0±0.0)%降低至(87.2±5.6)%,(66.2±4.8)%,(54.5±4.9)%,(42.5±6.4)%和(32.0±2.2)%(P<0.05,P<0.01);HeLa细胞G2/M期百分率逐渐增加,分别由对照组的(7.5±1.2)%增加到(12.9±1.2)%,(16.2±2.8)%,(23.6±3.9)%,(34.2±4.2)%和(52.6±7.8)%(P<0.05,P<0.01)。胡桃醌联合顺铂作用24 h,光镜下脱壁细胞增加,细胞形态变圆,较胡桃醌和顺铂单用组更加明显;联合用药组细胞存活率为(35.0±6.1)%,较胡桃醌和顺铂单用组〔(78.2±12.5)%和(58.5±10.9)%〕明显降低(P<0.01)。胡桃醌联合顺铂作用8 h,HeLa细胞早期凋亡率为(24.6±5.7)%,高于胡桃醌和顺铂单用组〔(6.7±1.5)%和(0.4±2.8)%〕(P<0.01)。结论胡桃醌可抑制HeLa细胞存活,促进HeLa细胞凋亡,与顺铂联合使用具有协同作用。     

定向胞内释放NO的半乳糖缀合物及其对HeLa细胞的抑制作用

目的研究糖基化NO供体-二醇二氮烯翁(d iazen ium-d iolate,NONOate)与β-半乳糖(β-galactose)的缀合物β-Gal-NONOate(β-galactosyl-d iazen iumd iolate)在肿瘤细胞中的NO释放及其对癌细胞的抑制效应。方法采用阳离子脂质体介导法,将真核表达载体pcDNA3-LacZ转染到HeLa细胞中;经G418(neomyc in)筛选,以邻-硝基苯-β-D-半乳糖苷(or-tho-n itrophenyl-beta-D-galactopyranoside,ONPG)法进行β-半乳糖苷酶(β-galactosidase)活性检测,获得稳定表达β-半乳糖苷酶的HeLa细胞株;通过G riess方法比较β-Gal-NONOate与NONOate在HeLa/LacZ+细胞和HeLa细胞中NO的释放量,根据细胞克隆形成率、细胞计数和MTT法比较β-Gal-NONOate与NONOate对细胞的作用。结果β-Gal-NONOate在HeLa/LacZ+细胞中NO的释放量和细胞毒性明显高于NONOate(P<0.05),而在未转染LacZ基因的HeLa细胞中比较稳定(P>0.05)。结论β-Gal-NONOate是一个成功的定向胞内释放的NO供体,可作为研究中的新型探针,并可能在抗肿瘤治疗中发挥作用。     

非酒精性脂肪性肝病患者血清脂联素水平及与酶学指标的相关性

目的通过检测非酒精性脂肪性肝病(NAFLD)患者血清脂联素及肝脏酶学等生化指标,探讨NAFLD患者血清脂联素水平与肝脏酶学等指标变化的相关性。方法选取60例NAFLD患者及60名健康对照者,用酶联免疫吸附试验法测定血清脂联素水平,结合患者的临床情况及肝脏酶学等指标,采用t检验和相关分析进行数据分析。结果NAFLD组血清脂联素水平为(12.7±2.8)mg/L,明显低于健康对照组(P<0.01);Spearman分析示脂联素与丙氨酸转氨酶(ALT)(r=-0.643,P<0.05)、天冬氨酸转氨酶(AST)(r=-0.472,P<0.05)、谷氨酰转肽酶(GGT)(r=-0.489,P<0.05)、甘油三酯(TG)(r=-0.671,P<0.05)、空腹血糖(FBG)(r=-0.710,P<0.05)、腰臀比(WHR)(r=-0.732,P<0.05)、体质量指数(BMI)(r=-0.719,P<0.05)呈负相关,与高密度脂蛋白胆固醇(HDL-C)(r=0.567,P<0.05)呈正相关。Logistic多因素回归分析显示脂联素与BMI(β=-0.314,P<0.05)呈独立负相关。结论NAFLD患者血清脂联素水平降低,其可能是NAFLD发病过程中起保护作用的脂肪细胞因子。     

脂联素、瘦素水平与冠心病及其相关危险因素研究

目的探讨血清脂联素、瘦素水平与冠心病及其相关危险因素的关系。方法拟诊冠心病的84例患者均行冠状动脉造影,冠心病组62例,对照组22例,检测其血清脂联素、瘦素水平并分析二者与冠心病危险因素的关系。结果脂联素水平冠心病组为(4.34±1.85)mg/l,低于对照组(8.94±2.59)mg/l;瘦素水平冠心病组为(7.88±2.20)ng/ml,高于对照组(2.73±2.15)ng/ml,差别均有统计学意义(P<0.05)。脂联素水平与体质指数(r=-0.360,P<0.01)、腰臀比(WHR)(r=-0.483,P<0.01)、空腹血糖(FBG)(r=-0.221,P<0.01)、甘油三酯(r=-0.258,P<0.05)、胰岛素抵抗指数(HOMA-IR)(r=-0.246,P<0.05)、超敏C反应蛋白(hs-CRP)(r=-0.341,P<0.05)、纤溶酶原激活物抑制物-1(r=-0.196,P<0.05)、瘦素(r=-0.175,P<0.05)成负相关,与高密度脂蛋白胆固醇(r=0.250,P<0.05)成正相关;瘦素水平与BMI(r=0.550,P<0.01)、总胆固醇(r=0.270,P<0.01)、纤溶酶原激活物抑制物-1(r=0.262,P<0.05)、HOMA-IR(r=0.186,P<0.05)、hs-CRP(r=0.155,P<0.05)成正相关。多元逐步回归分析进入脂联素回归方程的有:WHR、HOMA-IR、FBG、hs-CRP;进入瘦素回归方程的有:BMI、HOMA-IR。结论低脂联素水平、高瘦素水平可能与冠心病密切相关,二者可作为冠心病的独立危险因素。     

顺铂通过抑制转移抑制基因1-细胞外信号调节激酶及丝氨酸/苏氨酸激酶激活抑制HeLa细胞增殖

目的研究顺铂(DDP)抑制He La细胞增殖的分子机制。方法用顺铂0.02~75μmol·L-1处理He La细胞24或48 h,CCK-8法检测细胞存活,划痕实验检测细胞迁移力,流式细胞仪检测细胞周期,实时荧光定量PCR(q-PCR)检测转移抑制基因1(MTSS1)基因的表达,Western蛋白质印迹法检测磷酸化细胞外信号调节激酶(p-ERK)、磷酸化丝氨酸/苏氨酸激酶(p-AKT)和转移抑制基因1(MTSS1)蛋白水平。结果不同浓度DDP处理He La细胞24或48 h,可明显抑制He La细胞增殖(P<0.05),细胞的半数抑制浓度(IC50)值分别为4.14和11.82μmol·L-1;DDP处理组He La细胞较对照组迁移能力下降(P<0.05);DDP使He La细胞周期阻滞于S期;DDP抑制He La细胞MTSS1的基因表达,存在明显的浓度效应关系(r24 h=-0.965,P<0.01;r48 h=-0.953,P<0.01);p-ERK,p-AKT及胞内MTSS1蛋白表达水平均随DDP浓度增加显著下降(pERK:r24 h=-0.875,P<0.01;r48 h=-0.966,P<0.01;p-AKT:r24 h=-0.831,P<0.01;r48 h=-0.863,P<0.01;MTSS1:r24 h=-0.969,P<0.01;r48 h=-0.988,P<0.01)。结论 DDP可能通过下调MTSS1的表达水平并抑制ERK和AKT信号通路的激活而抑制宫颈癌He La细胞增殖和迁移。     

糖化血红蛋白与空腹血糖及餐后2 h血糖的相关性

目的探讨糖化血红蛋白(HbA1c)、空腹血糖(FPG)和餐后2 h血糖(2 h PG)的相关性。方法受试者为2010年1月至10月326例进行血糖和HbA1c检测的患者。血糖由葡萄糖氧化酶法测定,HbA1c由硼酸亲和层析法测定。结果 FPG与2 h PG显示了高度相关性,回归方程为Y=3.17+0.336X(r=0.866,P<0.01),当餐后2 h血糖为11.1 mmol/L时,FPG计算得6.9 mmol/L,当FPG为7.0 mmol/L时,餐后2 h血糖计算为11.08。且FPG和2 h PG血糖对于HbA1c的回归方程分别为Y=1.11X+0.1(r=0.809,P<0.01)和Y=2.97X-7.06(r=0.809,P<0.01),当HbA1c分别为6.0%和6.1%时,FPG分别计算得6.76 mmol/L和6.87 mmol/L,而2 h PG分别为10.71 mmol/L和11.07 mmol/L。结论 HbA1c与FPG和2 h PG之间有显著相关性,可以用HbA1c≥6.1%这个指标来筛查糖尿病患者。     

2型糖尿病患者血清铁蛋白与胰岛功能的相关性研究

目的分析血清铁蛋白(SF)与胰岛功能的相关性,进一步探讨铁储备在胰岛素抵抗及2型糖尿病发生、发展中的作用。方法研究对象分为糖尿病组与健康对照组,采用酶联免疫吸附试验(ELISA)检测SF水平,同时检测空腹胰岛素、血糖水平,并测量身高、体质量、血压、腰围、臀围,并计算腰臀比、体质量指数、胰岛素抵抗指数及胰岛β细胞功能(HOMA β-cell),分析其相关性。结果与对照组相比:糖尿病组患者SF水平显著升高(P<0.01);与胰岛素抵抗指数呈正相关(r=0.473,P<0.01);与HOMA β-cell呈负相关(r=-0.415,P=0.01)。结论铁超载可能通过增加胰岛素抵抗、损伤β细胞而引发糖尿病。     

健康体检人群体脂率与代谢指标的相关性研究

目的探讨深圳地区健康体检人群的体脂率与血压、血脂、血糖、尿酸、骨代谢等临床指标的相关性。方法回顾性分析2016年8~12月我院体检中心305例成年人健康体检者的体检资料,利用Pearson相关分析观察体脂率与各代谢指标的相关性。结果男性人群的体脂率与BMI(r=0.295,P<0.05)、TC(r=0.508,P<0.05)、LDL(r=0.576,P<0.05)、Ins(r=0.342,P<0.05)指标存在正相关;女性人群的体脂率与BMI(r=0.337,P<0.05)、TC(r=0.301,P<0.05)、Ins(r=0.299,P<0.05)、UA(r=0311,P<0.05)、Hcy(r=0.249,P<0.05)、Vit D(r=0.341,P<0.05)、β-CTX(r=0.357,P<0.05)、OC(r=0.464,P<0.05)指标均存在正相关,与HDL存在负相关(r=-0.325,P<0.05)。结论深圳地区成人健康体检者体脂率超重率高,体脂率可预测高脂血症、胰岛素抵抗、高尿酸血症、骨质疏松等疾病的发病风险,其性别亦为影响体脂率的重要因素,但无论对于男性、女性,均需采取针对性的健康教育及生活干预,以此稳定体脂率,保证其正常代谢,提高生活质量。     

慢性肾脏病患者血浆脑钠肽水平影响因素分析

目的 研究慢性肾脏病(CKD)患者脑钠肽(BNP)水平及其与临床和生化指标的相关性.方法 测定108例CKD患者血浆BNP水平,记录各患者临床和生化指标,分析BNP的影响因素.结果 Spearman相关分析结果提示,CKD患者BNP水平与年龄(r=0.272,P<0.01)、平均动脉压(MABP)(r=0.388,P<...     

端粒酶反义hTR和反义hTERT合用对宫颈癌HeLa细胞凋亡的影响

目的探讨端粒酶反义hTR和反义hTERT联合作用于宫颈癌HeLa细胞,对HeLa细胞凋亡的影响。方法实验分空白对照组、脂质体对照组、正义hTR组、正义hTERT组、反义hTR组、反义hTERT组及反义hTR+反义hTERT组。分别采用四甲基偶氮唑蓝法(MTT)、逆转录聚合酶链反应技术-端粒重复序列扩增(TRAP)酶联免疫法(ELISA)、吖啶橙染色法、流式细胞术检测宫颈癌HeLa细胞转染反义hTR和反义hTERT后细胞的端粒酶活性、形态学、凋亡和周期的改变。结果宫颈癌HeLa细胞转染端粒酶反义核酸后,出现明显的细胞凋亡形态学改变,端粒酶活性抑制率、凋亡率增高及细胞周期阻滞,与空白对照组、脂质体对照组及正义核酸组比较,统计学上差异有显著性(P<0.01)。HeLa细胞转染0.2μmol·L-1反义hTR+反义hTERT72h后,端粒酶活性抑制率、细胞凋亡率分别为80.5%、28.6%,与反义hTR组和反义hTERT组比较(端粒酶活性抑制率、细胞凋亡率分别为hTR:62.7%、13.2%;hTERT:66.5%、13.7%),差异有显著性(P<0.01,q值分别为0.919、1.075)。反义hTR+反义hTERT组比反义hTR组和反义hTERT组细胞凋亡的形态学改变更明显,反义hTR+反义hTERT组G0/G1期细胞比例增加(G1期细胞比例为57.8%,空白对照组G1期细胞比例为50.7%)。结论端粒酶反义核酸能通过抑制端粒酶活性诱导细胞凋亡,端粒酶反义hTERT和反义hTR有协同效应。     

三氧化二砷抗骨髓增生异常综合征骨髓血管新生机制的初步探讨

目的探讨骨髓增生异常综合征(MDS)患者骨髓是否存在血管新生及三氧化二砷(As2O3)的抗血管新生作用。方法①采用CD34单抗免疫组织化学染色法标记骨髓组织血管内皮细胞,盲法计数骨髓微血管密度(MVD)。②采用四甲基偶氮唑蓝方法观察As2O3对A组(HUVEC)及B组(经SKM-1细胞株上清液干预组HUVEC)的增殖抑制效应。结果①MDS患者骨髓微血管密度(MVD)显著高于正常对照组(P<0.01),并且随着病程的进展患者的MVD呈增高趋势(Rs=0.86 133,P<0.05);②As2O3对HUVEC及SKM-1细胞株干预组的作用均表现时间、剂量依赖性的增殖抑制效应(F=10.92,P<0.05);③HUVEC中加入SKM-1上清液共同培养对促进细胞生长有明显作用(P<0.01)。结论①MDS患者存在骨髓微血管密度增高,并与MDS患者的疾病进展正相关,血管新生在MDS的发生发展中起到重要作用;②As2O3以时间-剂量依赖性方式抑制HUVEC细胞的生长;③提示MDS细胞可分泌促进内皮细胞生长的细胞因子,通过旁分泌作用刺激内皮细胞生长。     

Decreased expression of human aquaporin-5 correlated with mucus overproduction in airways of chronic obstructive pulmonary disease

AIM: To investigate the relationship between aquaporin-5 (AQP5) expression and mucus overproduction in the airways of Chinese patients with chronic obstructive pulmonary disease (COPD) and the correlation with pulmonary function change. METHODS: Bronchial tissues were obtained from fiberoptic bronchoscopy and bronchial biopsy in West China Hospital from April to July 2004. Twenty-five patients were diagnosed as COPD patients, and another 20 were diagnosed as the control patients. The expressions of AQP5, mucin 5AC (MUC5AC), and mucin in bronchial tissues were detected by semiquantitative RT-PCR, in situ hybridization, and immunohistochemical and alcian blue-periodic acid-Schiff (AB-PAS) staining, respectively. RESULTS: Compared with the control group, an attenuated expression of AQP5 was detected throughout the bronchial tissues from patients with COPD (P<0.01), but no difference existed in the lung tissues (P>0.05). Simultaneously, increased staining of MUC5AC and mucus in submucosal glands were noted (P<0.01, respectively). Smoking attenuated the expressions of AQP5 and increased the staining of MUC5AC and mucus in submucosal glands in the COPD groups (P<0.01), while there were no significant differences observed in the control group (P>0.05). The decreased expression of AQP5 mRNA was correlated with forced expiratory volume in 1 s (FEV1)/forced vital capacity (FVC) (r=0.60, P<0.01), FEV1% predicted value (r=0.60, P<0.01), maximal expiratory flow in 50% vital capacity (V50) % predicted value (r=0.55, P<0.01), and maximal expiratory flow in 25% vital capacity (V25) % predicted value (r=0.45, P<0.01). The decreased expression was negatively correlated with MUC5AC mRNA of the epithelium airways (r=-0.45, P<0.01) and the AB-PAS-stained area of submucosal glands (r=-0.61, P<0.01). The upregulation of MUC5AC mRNA correlated with the positively AB-PAS-stained area of submucosal glands and correlated negatively with FEV1/FVC (r=-0.53; P<0.01), FEV1% predicted value (r=-0.53; P<0.01), V50% predicted value (r=-0.48; P<0.01), and V25% predicted value (r=-0.43; P<0.01). CONCLUSION: The attenuated gene expression of AQP5 existed in the airways of Chinese COPD patients, which was complicated by mucus hypersecretion. The decreased expression of AQP5 mRNA may be related to the severity of airflow obstruction.     

Bufadienolides from the Bufo viridis toad venom exert cytotoxic effects on cancer cells by inducing cell apoptosis and cell cycle arrest

A series of bufadienolides were isolated from the Bufo viridis toad venom, and their cytotoxic activities against three human cancer cell lines (HeLa, HT-29, MCF7) and a non-cancer cell line (L-O2) were explored using the MTT assay in vitro. All of nine compounds exhibited cytotoxic activities against the three cancer cell lines, with compound D4 exhibiting potent cytotoxic activity against HeLa cells and was better than positive control. Herein, we further evaluated the effect of compound D4 on HeLa cells. The results revealed that compound D4 has excellent cytotoxic effect on HeLa cells by inhibiting cell colony formation and migration, promoting cell apoptosis, increasing reactive oxygen species (ROS) levels and arresting of HeLa cells in S and G2/M phases. These findings encourage further work on the chemistry and bioactivity of the Bufo viridis toad venom.     

Effect of plasma lipid on pharmacokinetics of ciclosporin and its relationship with plasma prednisolone level in renal transplant patients

Ciclosporin (cyclosporine A, CyA) is a potent immunosuppressant used after organ transplantation. The pharmacokinetic properties of CyA vary widely and lipoproteins are the major complexing constituents for CyA in the plasma. Therefore, a change in lipoprotein level may influence the pharmacokinetic properties of CyA. Prednisolone (PSL) is concomitantly used with CyA as an immunosuppressant. After organ transplantation, hyperlipidaemia resulting from PSL therapy has been mostly observed and PSL increased the plasma lipoprotein level. Therefore, in this study, to obtain more useful information of the therapeutic drug monitoring (TDM) of CyA, the relationship between the plasma PSL level, plasma lipoprotein level and blood CyA level was investigated in detail. An open-label, non-randomized, retrospective study was performed. Data from 21 male and 11 female patients (age 11-65 years) who received a living-related renal transplantation from 2002 to 2004 were included. On postoperative days (PODs) 7, 14 and 28, the area under the plasma concentration-time curve until 9 h after 40 mg of PSL administration (AUCPSL40(0-9)) correlated well with total cholesterol (T-cho) (r=0.558, 0.768, 0.660, all P<0.05) and high-density lipoprotein (HDL) (r=0.688, P<0.05; 0.835, P<0.01; 0.508, p<0.05), and correlated negatively with very-low-density lipoprotein (VLDL) (r=-0.486, p<0.01; -0.776, p<0.01; -0.967, p<0.01). In addition, AUC until 9 h after CyA administration (AUCCyA0-9) also correlated with T-cho (r=0.797, p<0.01; 0.577, p<0.05; 0.901, p<0.01), HDL (r=0.514, p<0.05; 0.614, p<0.05; 0.893, p<0.01) and low-density lipoprotein (LDL) (r=0.906, p<0.01; 0.573, p<0.05; 0.537, p<0.05), and there was a negative correlation with VLDL (r=-0.480, -0.630, -0.632, all p<0.05). Moreover, AUCCyA0-9 correlated well with AUCPSL40(0-9) (r=0.728, p<0.01; 0.482, p<0.05; 0.688, p<0.05); namely, it was considered that the variety of plasma PSL concentrations influenced the pharmacokinetic properties of CyA through the change in lipoprotein levels. These results suggested that monitoring of the biochemical parameters of the plasma lipid and plasma PSL level might be useful for the TDM of CyA.     

Bersaldegenin-1,3,5-orthoacetate induces caspase-independent cell death,DNA damage and cell cycle arrest in human cervical cancer HeLa cells

ContextBufadienolide compounds occur in many plants and animal species and have strong cardiac and anti-inflammatory properties. The compounds have been recently investigated for cytotoxic and antitumor activity.ObjectiveThe cytotoxic effect of bersaldegenin-1,3,5-orthoacetate – a bufadienolide steroid occuring in plants from Kalanchoe genus (Crassulaceae), was evaluated with cervical cancer HeLa cells in vitro.Materials and methodsThe cytotoxic activity of the compound (at 0.1–20.0 μg/mL) on the cells was determined by Real-Time Cell Analysis (RTCA) system for 24 h. The estimation of cell cycle arrest, reactive oxygen species (ROS) production, reduction of mitochondrial membrane potential (MMP), and caspases-3/7/9 activity in the HeLa cells treated with the compound was done by flow cytometry and luminometric technique. DNA damage in the cells was estimated by immunofluorescence staining and the comet assay with etoposide as a positive control.ResultsThe compound had strong effect on the cells (IC₅₀ = 0.55 μg/mL) by the suppression of HeLa cells proliferation in G2/M phase of cell cycle and induction of cell death through double-stranded DNA damage and reactive oxygen species overproduction. Furthermore, we did not observe an increase in the activity of caspase-3/7/9 in the treated cells as well as a decrease in cellular mitochondrial membrane potential. Gene expression analysis revealed the overexpression of NF-Kappa-B inhibitors genes (>2-fold higher than control) in the treated cells.ConclusionsBersaldegenin-1,3,5-orthoacetate induces cell cycle arrest and caspase-independent cell death through double-stranded DNA damage. These results are an important step in further studies on cell death signalling pathways induced by bufadienolides.     

不伴精神症状的苯丙胺类兴奋剂依赖者家庭环境因素的初步分析

目的:初步分析不伴精神症状的苯丙胺类兴奋剂依赖者的家庭环境。方法:选择60例苯丙胺类兴奋剂依赖者自评中文版的家庭环境量表(FES-CV)。结果:该类苯丙胺类兴奋剂依赖者的家庭环境量表中亲密度、矛盾性、娱乐性、控制性等因子分均高于国内常模,差异有显著性(分别:t=2.543,P=0.014;t=2.361,P=0.022;t=2.186,P=0.033;t=2.571,P=0.013);情感表达、成功性、知识性等因子分低于国内常模,差异有显著性(分别:t=-2.213,P=0.031;t=-2.631,P=0.011;t=-2.268,P=0.027);同时,亲密度与控制性呈正相关(r=0.308,P<0.05),情感表达与组织性呈正相关(r=0.430,P<0.01),矛盾性与独立性、知识性、娱乐性、道德宗教观、控制性呈正相关(r=0.271,P<0.05;r=0.512,P<0.01;r=0.493,P<0.01;r=0.425,P<0.01;r=0.413,P<0.01),独立性与知识性、道德宗教观、控制性呈正相关(r=0.372,P<0.01;r=0.370,P<0.01;r=0.374,P<0.01),知识性与娱乐性、道德宗教观、控制性呈正相关(r=0.805,P<0.01;r=0.698,P<0.01;r=0.648,P<0.01),娱乐性与道德宗教观、控制性呈正相关(r=0.637,P<0.01;r=0.691,P<0.01),控制性与道德宗教观呈正相关(r=0.501,P<0.01)、与组织性呈负相关(r=-0.381,P<0.01)。结论:不伴精神症状的苯丙胺类兴奋剂依赖者具有高亲密度、低情感表达、高矛盾性、低成功性、低知识性、高娱乐性和高控制性的家庭环境特点。建议结合相关性研究制定针对家庭环境变化的家庭综合干预的心理治疗,有助于尽快康复。