叶绿酸联合赛特铂对人SMMC-7721肝癌细胞株影响的初步研究

目的:探讨叶绿酸(CHL)联合赛特铂(JM216)对人SMMC-7721肝癌细胞凋亡及增殖的作用。方法:选用CHL(1×10-5 mol/L)及不同浓度JM216(10、20和40mg/L),1×10-5 mol/L的CHL分别联合不同浓度JM216作用于SMMC-7721肝癌细胞24、48和72h后,用MTT比色法观察其对肝癌细胞的生长抑制作用,并观察细胞形态变化;用流式细胞术分析药物作用48h时肝癌细胞的周期分布和凋亡情况。结果:CHL组、不同浓度JM216组和联合组细胞生长抑制率均高于对照组,联合组1和联合组2以及联合组3的细胞生长抑制率均高于相应浓度的JM216组。CHL组SMMC-7721细胞周期中G0/G1期比例为(55.94±3.49)%,高于对照组的(50.52±2.56)%,差异有统计学意义,P<0.05;而JM216组SMMC-7721细胞周期中S期比例为(45.12±1.42)%,高于对照组的(27.60±1.27)%,差异有统计学意义,P<0.05。CHL组和JM216组细胞凋亡率分别为(13.48±1.63)%和(15.51±2.09)%,均高于对照组的(1.61±0.47)%,差异有统计学意义,P<0.05;联合组细胞凋亡率为(30.22±3.20)%,高于JM216组和对照组,差异均有统计学意义,P<0.05。结论:CHL和JM216均可抑制肝癌SMMC-7721细胞的生长并诱导其凋亡,且联用后作用增强。     

CCR4对肝癌细胞索拉非尼放射敏感性及裸鼠成瘤的影响

目的 探讨CC趋化因子受体4(CCR4)对肝癌细胞索拉非尼放射敏感性及裸鼠成瘤的影响。方法 采用Western blot检测CCR4在肝癌细胞系中的表达。利用慢病毒感染构建过表达及敲减CCR4的PLC/PRF/5和SMMC-7721稳转株,并采用Western blot进行验证。采用平板克隆形成实验检测CCR4对肝癌细胞放射敏感性的影响。通过裸鼠成瘤实验检测CCR4对肝癌细胞体内成瘤的影响。结果 高转移肝癌细胞中CCR4呈高表达,低转移肝癌细胞中CCR4呈低表达。成功构建稳定过表达CCR4的PLC/PRF/5和敲减CCR4的SMMC-7721肝癌稳转株,过表达CCR4可逆转索拉非尼放疗对PLC/PEF/5的抑制作用,而敲减CCR4可增加SMMC-7721对索拉非尼放射敏感性。过表达CCR4可促进PLC/PEF/5裸鼠体内成瘤能力,而敲减CCR4可抑制SMMC-7721裸鼠体内成瘤能力。结论 CCR4高表达能显著增强肝癌PLC/PEF/5细胞对索拉非尼放射抵抗及裸鼠体内成瘤能力,而敲减CCR4则可显著增加肝癌SMMC-7721细胞对索拉非尼放射敏感性并抑制其裸鼠体内成瘤能力     

鱼藤素对肝癌细胞株SMMC-7721增殖抑制和凋亡诱导作用的研究

目的 研究鱼藤素对肝癌细胞株SMMC-7721增殖和凋亡的影响及其作用机制.方法 MTT法检测鱼藤素对SMMC-7721细胞增殖的抑制作用;DNA琼脂糖凝胶电泳观测细胞凋亡情况;采用Western blotting法检测相关蛋白表达水平的变化,探讨其作用机制.结果 鱼藤素呈剂量和时间依赖性抑制SMMC-7721细胞增殖,诱导细胞凋亡致DNA ladder形成,出现相应的形态学改变;鱼藤素可明显降低Akt的磷酸化水平.结论 鱼藤素可通过下调Akt的活性,抑制SMMC-7721的增殖并诱导细胞凋亡,或可成为一种新的抗肝癌药物.     

lncRNA PTPRG-AS1通过靶向miR-124-3p调控肝癌细胞凋亡和放射敏感性的研究机制

目的:探讨lncRNA PTPRG-AS1对肝癌细胞凋亡和放射敏感性的影响及作用机制。方法:培养正常肝细胞L02和肝癌细胞HepG2、SMMC-7721和BEL-7402,qRT-PCR检测细胞中PTPRG-AS1和miR-124-3p表达水平。转染si-PTPRG-AS1、miR-124-3p mimics至HepG2细胞,抑制HepG2细胞中PTPRG-AS1表达或过表达miR-124-3p,流式细胞术检测细胞凋亡,克隆形成实验检测细胞放射敏感性,Western Blot法检测Bcl-2和Bax蛋白表达。生物信息学软件预测PTPRG-AS1与miR-124-3p存在互补的核苷酸序列,双荧光素酶报告基因实验验证PTPRG-AS1与miR-124-3p之间的调控关系。结果:与正常肝细胞L02相比,肝癌细胞HepG2、SMMC-7721和BEL-7402中PTPRG-AS1表达显著升高（P＜0.05）,miR-124-3p表达显著降低（P＜0.05）。抑制PTPRG-AS1或过表达miR-124-3p均可促进肝癌HepG2细胞的凋亡,增强肝癌HepG2细胞的放射敏感性,抑制Bcl-2蛋白表达,促进Bax蛋白表达。PTPRG-AS1负调控miR-124-3p表达。抑制miR-124-3p表达可部分逆转抑制PTPRG-AS1表达对肝癌HepG2细胞凋亡的促进作用以及放射敏感性的增强作用。结论:抑制PTPRG-AS1表达可能通过上调miR-124-3p表达促进肝癌细胞的凋亡并提高其放射敏感性,是肝癌治疗的潜在作用靶点。     

沙利度胺对人肝癌细胞株SMMC-7721体外生长的抑制作用

目的 研究沙利度胺对人肝癌细胞株SMMC-7721体外生长的抑制作用及其可能的机制.方法 将不同浓度的沙利度胺作用于人肝癌细胞株SMMC-7721,采用四甲摹偶氮唑蓝(MTT)法检测沙利度胺对SMMC-7721细胞的增殖抑制作用.将SMMC-7721细胞培养至对数生长期,采用DNA琼脂糖凝胶电泳、荧光显微镜观察、流式细胞仪检测等方法 观察沙利度胺处理后SMMC-7721细胞的凋亡梯度、形态学变化和凋亡率,并对凋亡调控蛋白caspase-3的表达进行测定.采用酶联免疫吸附(ELISA)法测定不同浓度的沙利度胺处理后SMMC-7721细胞表达血管内皮生长因子(VEGF)的变化.结果 沙利度胺的浓度从3.125μg/ml增至200μg/ml时,其对SMMC-7721细胞的增殖抑制率从11.7%增至34.2%;当沙利度胺的浓度>25 μg/ml时,其对SMMC-7721细胞的增殖抑制作用明显强于空白对照组(P<0.05).200 μg/ml的沙利度胺处理SMMC-7721细胞24 h后,行琼脂糖凝胶电泳,可见到DNA梯形条带;48 h后梯形条带更明显,并且在荧光显微镜下可见SMMC-7721细胞出现核固缩和核裂解现象.200μg/ml的沙利度胺处理SMMC-7721细胞12、24、48和72 h时,碘化丙啶(PI)法检测SMMC-7721细胞的凋亡率分别为3.1%±0.5%、8.4%±1.3%、19.4%±3.5%和25.8%±2.1%,24 h起的凋亡率均明显高于空白对照组SMMC-7721细胞48 h的自然凋亡率(1.6%±0.6%,均P<0.05).50、100和200μg/ml的沙利度胺处理SMMC-7721细胞48 h时,Annexin V-FITC/PI双标法检测SMMC-7721细胞的凋亡率分别为8.7%±1.2%、16.8%±2.5%和25.4%±4.5%,均明显高于空白对照组SMMC-7721细胞48 h的自然凋亡率(2.1%±0.5%,均P<0.05).随着沙利度胺浓度的增加,表达caspase-3蛋白的SMMC-7721细胞数量不断增加,而SMMC-7721细胞中VEGF的含量却逐渐下降.结论 沙利度胺可能通过诱导肝癌细胞的凋亡、抑制肿瘤血管的生成而发挥双重抗肿瘤生长的作用.     

紫杉醇诱导肝癌细胞SMMC—7721凋亡的实验研究

目的：研究紫杉醇（Taxol)对肝癌细胞SMMC-7721的生长抑制和诱导凋亡的作用。方法：应用MTT法观察紫杉醇对SMMC-7721的生长抑制作用。通过瑞氏染色，荧光显微镜，透射电镜，流式细胞仪和TUNEL染色研究紫杉醇对SMMC-7721诱导凋亡的作用。结果：紫杉醇对肝癌细胞SMMC-7721有明显的生长抑制作用，其IC50为21.6nmol/L。紫杉醇作用细胞后，可看到较为典型的细胞凋亡的形态学变化。细胞核固缩，染色质凝集成新月型紧贴于核膜周边，核碎裂，染色质片断化，凋亡小体形成等，流式细胞仪上可见S期和G2/M期和DNA含量增加，并出现典型的亚二倍体“凋亡小峰”。TUNEL染色显示。紫杉醇作用于SMMC-7721后，随药物浓度增加和作用时间延长，凋亡指数增加。结论：紫杉醇对SMMC-7721有明显的生长抑制作用；紫杉醇对SMMC-7721有诱导凋亡的作用。     

人肝癌细胞BEL-7402非常规照射模式生物效应探讨

目的 探讨不同照射模式对人肝癌细胞株BEL-7402生物效应变化影响及其与人肝细胞株QSG-7701放射敏感度差异。方法 6MV X线两种模式照射BEL-7402细胞(包括0、1、2、3、5、7、9、10Gy共8个吸收剂量点,剂量率3Gy/min)：(1)急速照射组,照射完成时间0~4min;(2)模拟三维适形照射组：照射完成时间：12~15min。成克隆分析法计算存活分数,多靶单击数学模型拟合曲线,求出SF2、D0、Dq等参数值;6MV X线单次5Gy照射体外培养的BEL-7402、QSG-7701 细胞, 采用MTT 比色法、流式细胞术测定细胞受照后不同时间存活率、凋亡率、细胞周期。结果 BEL-7402细胞模拟急速照射组和三维适形照射组D0、Dq、SF2值分别为1.78和1.69、1.45和1.54、0.625和0.654;QSG-7701 细胞照后24 h 凋亡率最高(18 %),而BEL-7402细胞照后12h凋亡率最高(32 %),存活率最低(65%),两株细胞照后12 h 凋亡率和存活率差异最大（P<0.01）。结论 三维适形照射模式下分次照射时间延长,生物效应下降;BEL-7402及QSG-7701两株细胞在照后存在放射敏感度差异。     

应用中空纤维法评价酪丝缬肽对肝癌的抑制作用

目的 观察三肽化合物酪丝缬肽(YSV)的抗肝癌作用.方法 应用新型四氮唑盐(MTS)法,检测YSV对BEL-7402、SMMIC-7721、Hep3B、Her,G2和SK-HEP-1人肝癌细胞体外生长的抑制作用;应用中空纤维管皮下移植模型,进一步观察YSV对这5种人肝癌细胞体内生长的影响.结果 0.1～1.6 mg/ml的YSV能够明显抑制BEL-7402、SMMC-7721、Hep3B、ItepG2和SK-HEP-1人肝癌细胞的体外生长,YSV组的吸光度(A)值与相应阴性对照组相比,差异均有统计学意义(P<0.05),其中YSV对人肝癌细胞BEL-7402的抑制作用最强,1.6 mg/ml的YSV抑制率达63.3%.160μg·kg-1·d-1和320 μg·kg-1·d-1的YSV均能够明显抑制BEL-7402、SMMC-7721、ttep3B、HepG2和SK-HEP-1人肝癌细胞的体内生长,YSV组的A值与相应生理盐水组相比,差异均有统计学意义(P<0.05).YSV对人肝癌细胞BEL-7402的抑制作用最强,320 μg·kg-1·-1 YSV的抑制率为53.1%.结论 中空纤维法可以快速、准确地观察药物对不同肝癌细胞株体内生长的抑制作用,为YSV的临床应用提供了实验指导和可靠证据.     

STAT3抑制剂S3I-201对人肝癌BEL-7402细胞增殖、凋亡及细胞周期的影响

目的 研究STAT3小分子抑制剂S3I-201对人肝癌细胞系BEL-7402增殖、凋亡与细胞周期的影响和初步的作用机制。方法 分别采用细胞活性检测试剂盒cell couting kit-8(CCK-8)和EdU细胞增殖法检测S3I-201对人肝癌细胞BEL-7402细胞增殖的影响,应用流式细胞术分析S3I-201对BEL-7402的细胞凋亡和周期的影响,通过Western blotting检测凋亡相关蛋白Bcl-2、Bax的表达。结果 CCK-8及EdU结果显示,S3I-201能抑制BEL-7402细胞的生长和增殖(P<0.05),使BEL-7402细胞阻滞于S期,具有诱导细胞凋亡的作用,上调Bax的表达,下调Bcl-2的表达(P<0.05)。结论 STAT3小分子抑制剂S3I-201通过促进BEL-7402凋亡、上调促凋亡蛋白Bax、下调抗凋亡蛋白Bcl-2的表达,并将BEL-7402阻滞于S期而发挥抗肿瘤效应。     

极低频磁场对肝癌细胞株BEL-7402的作用初步观察

目的:观察极低频磁场对人肝癌细胞株BEL-7402的作用,探讨其抑制细胞生长的机制.方法:建立极低频磁场生物模型.极低频磁场处理(100 Hz,0.7 mT,30 min)人肝癌细胞系BEL-7402后,给予不同剂量X射线照射(2、4、6、8和10 Gy).作细胞集落形成试验;流式细胞仪研究极低频磁场作用下人肝癌细胞系BEL-7402细胞周期及相关蛋白的改变;RT-PCR检测p53水平.结果:细胞集落形成试验结果显示,极低频磁场加X射线组抑制率为73.00%、78.00%、58.00%、55.00%和67.00%,单纯X射线组抑制率为38.00%、41.00%、20.00%、24.00%和30.00%,F=5.89,P=0.03.流式细胞仪检测显示,5种不同放射强度下,极低频磁场加放射组凋亡率为10.00%、14.50%、4.30%、5.10%和7.10%,单纯放射组凋亡率为0.10%、8.10%、0.10%、0.41%和2.20%,极低频磁场加X射线对肝癌细胞株BEL-7402的凋亡率均高于单纯放射,2和4 Gy照射时最为明显,两组差异有统计学意义,F=6.9,P=0.03.极低频磁场组细胞生长停滞于G2/M期,细胞周期停滞相关的p53蛋白水平均显著增高.结论:100 Hz,0.7 mT极低频磁场能诱导人肝癌细胞BEL-7402产生凋亡.极低频磁场抑制癌细胞增殖作用的机制与G2/M期停滞有关.其中p53激活是引起细胞周期停滞重要原因.     

Ringworm in animals in a farm in Assiut

Bericht über das Auftreten von Hautmykosen bei Tieren auf der Farm der Landwirtschaftlichen Fakultät der Universität Assiut. Von 70 Kälbern waren 42 pilzkrank, davon 22 durch T. mentagrophytes, 20 durch T. verrucosum. Von 180 Kühen waren 8 pilzbefalien, sämtlich durch T. mentagrophytes. Von 2 kranken Bullen wurde T. verrucosum isoliert. Auch die 3 Pferde und 2 Maultiere der Farm waren pilzinfiziert; Erreger war in diesen Fällen T. equinum. Auch ein Kalb war von T. equinum befallen. Die gleiche Pilzart wurde ferner von 3 Tierpflegern isoliert, die Pilzherde am Hals und an den Armen aufwiesen.     

Statistical challenges in preprocessing in microarray experiments in cancer

Many clinical studies incorporate genomic experiments to investigate the potential associations between high-dimensional molecular data and clinical outcome. A critical first step in the statistical analyses of these experiments is that the molecular data are preprocessed. This article provides an overview of preprocessing methods, including summary algorithms and quality control metrics for microarrays. Some of the ramifications and effects that preprocessing methods have on the statistical results are illustrated. The discussions are centered around a microarray experiment based on lung cancer tumor samples with survival as the clinical outcome of interest. The procedures that are presented focus on the array platform used in this study. However, many of these issues are more general and are applicable to other instruments for genome-wide investigation. The discussions here will provide insight into the statistical challenges in preprocessing microarrays used in clinical studies of cancer. These challenges should not be viewed as inconsequential nuisances but rather as important issues that need to be addressed so that informed conclusions can be drawn.     

Trends in zygomycosis in children

Zygomycosis, or mucormycosis, is associated with significant morbidity and mortality in both children and adults. Studies in adults have shown an increase in the incidence of zygomycosis, particularly among haemtopoietic stem cell transplant (HSCT) recipients and patients with haematologic malignancies. There is a paucity of data on the epidemiology of zygomycosis in children. We performed a retrospective analysis to describe trends in zygomycosis between 1 January 2003 and 31 December 2010. We used the Pediatric Health Information System (PHIS) database to identify paediatric patients who were diagnosed with zygomycosis during the study period. Administrative data on diagnoses, demographics, underlying conditions and clinical experiences were collected. Summary statistics were calculated and tests for trend were conducted. We identified 156 unique patients with zygomycosis. The prevalence of zygomycosis did not significantly increase over time (P=0.284). The most common underlying condition was malignancy (58%) and over half received intensive care. Voriconazole utilisation among all hospitalised children significantly increased during the period (P=0.010). Our study demonstrates that the incidence of zygomycosis is not significantly increasing. During the time period there was a significant increase in the use of voriconazole among children.     

Dermatomycosis in Goats in India

Summary: In an extensive survey involving 2176 goats 1.56% of goats manifested clinical lesions of ringworm infection. Animals below the age of 6 months were affected most (4.20%). The incidence of infection was higher during the winter months. T. verrucosum, T. mentagrophytes and M. gypseum were isolated from the skin scrapings of 12, 4, and 2 goats respectively. It is suggested that the infected animals could be a source of disease to human population in which the zoophilic dermatophyte invasion is characterized by severe inflammatory lesions of the skin. The zoonotic importance of different dermatophytes is stressed.
Zusammenfassung: In einer ausgedehnten Untersuchung an 2176 Ziegen wurden bei 1,56% klinische Zeichen einer Hautmykose festgestellt. Tiere, die jühger als 6 Monate alt waren, zeigten mit 4,20% am häfigsten Krankheitssymptome. In den Wintermonaten war die Erkrankungshäufigkeit am gröBten. T. vermcosum, T. mentagrophytes and M. gypseum wurden jeweils von 12,4 bzw. 2 Ziegen isoliert. Die inflzierten Tiere köinnen eine Infektionsquelle für die menschliche Bevölkerung darstellen und dort Mykosen mit stark entzündlichen Veränderungen auslösen. Die Bedeutung verschiedener Dermatophyten als Erreger von Zoonosen wird hervorgehoben.     

Hemoglobin in lymphogranulomatosis in children

The peculiarities of hemoglobin content and properties in lymphogranulomatosis were studied in children by different methods. In most patients the content of fetal hemoglobin and methhemoglobin was found to increase and the rate of ferricyanide oxidation of hemoglobin also increases. Simultaneously, the rate of non-homogenicity of basic hemoglobin fractions was observed to increase on account of immunologically different components.     

Sex hormones in women in rural China and in Britain

Plasma concentrations of certain hormones linked to breast cancer risk were measured in age-pooled samples from 3,250 rural Chinese women in 65 counties, and 300 British women, all aged 35-64. In age-groups 35-44, 45-54 and 55-64 respectively, mean oestradiol concentrations were 36% (P = 0.043), 90% (P less than 0.001) and 171% (P = 0.001) higher in the British than in the Chinese women, and mean testosterone concentrations were 48% (P less than 0.001), 68% (P less than 0.001) and 53% (P = 0.001) higher in the British than in the Chinese women. The difference in testosterone concentrations between the two countries appeared to be due largely to the lower average body weight in the Chinese women. Sex hormone binding globulin did not differ significantly between the two countries in age groups 35-44 and 45-54, but was 15% (P = 0.002) lower in the British than in the Chinese women at ages 55-64. Prolactin concentrations did not differ significantly between the two countries in any age group.