Expressions of Nucleoside Diphosphate Kinase （nm23） in Tumor Tissues are Related with Metastasis and Length of Survival of Patients with Hepatocellular Carcinoma

Objective To evaluate the relationship of expressions of nucleoside diphosphate kinase （nm23） and proliferating cell nuclear antigen （PCNA）, as well as apoptosis, with the prognosis of HCC patients by analyzing their pathological and clinical data. Methods The expressions of nm23 and PCNA were analyzed by immunohistochemistry and the apoptotic phenomena were detected by TUNEL technique in the liver samples from 43 HCC tissues, 39 para-neoplastic tissues, and 10 normal tissues. The mean apoptosis index （AI） and proliferative index （PI） in individual sample were calculated. Results As shown by the detection, 32.6% of carcinomas had negative nm23 signal in tumor tissues, whereas all para-neoplastic and normal tissues had positive nm23. The AI in nm23 positive HCC was significantly higher than that in nm23 negative one, with statistical difference （P0.05）. Furthermore, the expressions of nm23, and the values of AI and PI were contrastively analyzed with some main pathological and clinical data of HCC. It revealed that HCC with extrahepatic metastasis showed remarkable correlation with the negative nm23 （P=0.013） and higher PI values of HCC （P=0.015）. The disease-free survival in HCC patients with negative nm23 expression was significantly poorer than that in patients with positive nm23 expression. Conclusion These data suggest that expressions of nm23 protein in tumor tissues are correlated with occurrences of metastasis and length of survival of the HCC patients, which may be an indicator for their prognosis.     

Expressions of SE-1, CD31 and CD105 in the vascular endothelial cells and serum of rat with hepatocellular carcinoma

Background Hepatocellular carcinoma （HCC） is one of the deadliest cancers worldwide. In order to investigate the molecular biologic mechanism of HCC＇s development, we studied the expressions of SE-1, CD105 and CD31 in tumor endothelial cells （TECs） of HCC and in the serum of rats. Methods We analyzed the expressions of SE-1, CD31 and CD105 in rat HCC tumor tissues using immunohistochemistry （IHC）. Twenty HCC bearing rats and eighteen normal rats were examined for the expressions of SE-1, CD31 and CD105 antigens in serum by enzyme-linked immunosorbent assay （ELISA）. Results SE-1, CD31 and CD105 antigens were detected both in HCC tissue and in normal liver tissue with higher expressions of CD31 and CD105 in HCC while the SE-1 antigen expression was higher in normal liver. Similarly, serum CD31 and CD105 in rats with HCC were significantly increased compared with normal rats （t=-2.8628, P=0.0086; t=4.4922, P 〈0.0001, respectively）. In contrast, SE-1 antigen in HCC rat serum was significantly decreased compared with normal rats （t=3.4983, P=-0.0011）.Conclusion SE-1, CD31 and CD105 are closely related with liver tumor angiogenesis, which is similar to their performances in terms of their expressions in the serum.     

Expression and Prognostic Significance of Livin in the Progression of Bladder Cancer

It has been suggested that progression of bladder transitional cell cancer (BTCC) may be regulated at the molecular level by a typical pattern of expression of genes involved in apoptosis. Re-cently Livin, belonging to the inhibitors of apoptosis (IAP) family, has been found to be expressed in most solid tumors, where its expression is suggested to have clinical significance. In order to explore the significance of Livin expression in the development of BTCC, immunohistochemistry and RT-QPCR were used to detect the expression of Livin mRNA in tumor tissues and adjacent normal tissues of 30 cases of BTCC. The results showed that the positive rate of Livin expression in adjacent normal tissues and tumor tissues was 0 and 60% (18/30) respectively. The -△△CT value of Livin in BTCC tissues was 8.0454 (7.4264-8.6644) times of that in adjacent normal tissues. The expression of Livin mRNA had no correlation with tumor pathological grades and clinical stages. It was sug-gested that there was weak expression of Livin mRNA in adjacent normal tissues, but strong in tumor tissues.     

Expression of p53 isoforms in renal cell carcinoma

Background Several isoforms of p53 have been reported, which may have varying functions and expressions. This study aimed to analyze the expression patterns of p53 isoforms in renal cell carcinoma （RCC） at the mRNA and protein levels and their associations with clinical and pathologic factors to explore the mechanism of p53 isoforms＇ activity in RCC. Methods The specimens of tumours （T） and clinically normal tissues （N） adjacent to them were collected from 41 patients with RCC. mRNA expression levels of p53 isoforms were detected using RT-PCR followed by nested PCR. Protein expression levels were detected using immunohistochemisty and Western blotting with the anti-p53 antibodies DO-1 and DO-12. The data were analyzed with clinicopathological features by X^2 test or Fisher＇s exact test. Results p53 mRNA was expressed in all tumours and matched clinically normal tissue adjacent to the tumour. All six isoforms could be detected in tumour and normal tissues, with the exception of the △133p53β isoform, which was not detected in the normal tissue. Of the six isoforms, p53β mRNA was significantly overexpressed in tumour samples （P 〈0.001）, and correlated with tumour stage. Nested PCR results consistently indicated the presence of p53γ （19T/22N）, △133p53 （33T/26N）, △133p53β （2T/0N）, and △133p53γ （13T/9N）. Immunohistochemical analysis showed that p53 was expressed only in tumour tissues and correlated with tumour stage and grade. The results of Western blotting analysis were consistent with these findings. Conclusions Although all six isoforms are present in RCC, their function in tumour development or progression might be different. Our findings suggest that p53β might play an important role in the formation of RCC and it might be used as a new predictor and therapeutic target for RCC.     

Relationship between Cytosol Estrogen receptor Level in Uterine Leiomyoma and Sex Steroid Level in Plasma,Leiomyoma and Myometrium

This study was designed to evaluate the effect of estrogen on thegrowth of leiomyomas and the mechanism of testosterone propionate action on thetumor.The levels of estrogen receptors in uterine tissues and of estradiol andprogesterone in uterine tissues and plasma were determined in 47 women withleiomyoma.The levels of estrogen receptor and estradiol in leiomyomatous tissueswere 37.6±4.0 fmol/mg protein and 401.7±92.6 pg/g tissue respectively.Both ofthem were higher than the corresponding values in normal uterine tissues(P<0.05).The concentration of estrogen receptor in the leiomyomas of 11 patientstreated with testosterone propionate was 18.9±9.4 fmol/mg protein,significantlylower than that of the untreated group(P<0.05).It is suggested that the changesof the levels of estrogen receptor and estradiol in the leiomyomatous tissues berelated to the growth of the tumor and one of the ways for testosterone propionateto inhibit the growth of leiomyomas might be due to the suppression of theestrogen receptor content in leiomyomas.     

XIAP as a prognostic marker of early recurrence of nonmuscular invasive bladder cancer

Background Dysregulation of apoptosis has been implicated not only in carcinogenesis and tumor progression but also in tumor recurrence. We investigated whether the expression of X-linked inhibitor of apoptosis （XIAP） might predict early recurrence in patients with non-muscular invasive bladder cancer. Methods The cohort comprised 176 consecutive patients with primary superficial bladder cancer treated with transurethral resection. Immunohistochemical staining using the standard avidin-bioUn-peroxidase technique and RT-PCR were used to detect XIAP protein and mRNA expressions in cancer tissues. The relationship between XIAP expression and clinicopathological characteristics, cancer recurrence were analyzed. Results XIAP expression was observed in 108 cases （61.4%） and no expression in 68. There was no correlation between XIAP expression rate and the tumor pathological grade, but was an apparent trend toward the increased XIAP levels from well （G1） to poor （G3） differentiated cancer. Eighty-two （46.6%） patients experienced tumor recurrence at a mean of 28.6 months of the follow-up; 66 of them expressed XIAP （61.1%） and 16 were XIAP negative （23.5%）. Twelve patients presented with invasive disease at the time of relapse and all of them expressed XIAP. Patients without XIAP expression or with low tumor grades had significantly higher recurrence-free survival than those with XIAP expression （log rank test ,P=-0.0015） or high tumor grades （log rank test P〈0.001）. Multivariate analysis revealed that XIAP expression, tumor grade, and tumor number were independent predictors for the recurrence of non-muscular invasive bladder cancer （P=-0.004, 0.016, and 0.043, respectively）. Conclusions XIAP may be considered as a new independent prognostic marker for early recurrence of non-muscular invasive bladder cancer.     

凋亡抑制蛋白PED/PEA-15和XIAP在肝细胞肝癌中的表达及意义

目的 探讨凋亡抑制蛋白PED/PEA-15和XIAP在肝细胞肝癌中的表达及临床病理因素的关系.方法 我们应用RT-PCR和Western印迹检测了40例肝细胞肝癌和相应癌旁组织及12例正常肝组织中PED/PEA-15和XIAP mRNA及蛋白的表达,并结合临床病理特征进行分析.结果 肝细胞肝癌组织中PED/PEA-15和XIAP mRNA蛋白的表达均明显高于癌旁组织和正常肝组织(相对吸光度比值为0.636±0.061、0.352±0.068、0.179±0.036与0.579±0.090、0.344±0.084、0.184±0.038)(P均＜0.01),PED/PEA-15和XIAP mRNA和蛋白的表达在不同年龄、性别、肿瘤大小、肿瘤数目、门静脉转移和肿瘤复发间的差异无统计学意义(P均＞0.05),在不同的病理分级和临床分期间差异有统计学意义(P＜0.05).结论 凋亡抑制蛋白PED/PEA-15和XIAP在肝细胞肝癌中呈高表达,可作为判断人肝细胞肝癌生物学行为的参考指标且有望成为肝细胞肝癌靶向基因治疗的新靶点.     

人白细胞介素15基因克隆及其在大肠杆菌中的表达

目的 克隆人白细胞介素（IL）15全长cDNA,并在大肠杆菌中表达。方法从人外周血分离的淋巴细胞中提取总RNA,通过RT—PCR扩增出hIL-15全长cDNA。构建到原核表达载体pET28a（＋）中,通过卡那霉素平板筛选及PCR鉴定,挑出阳性克隆进行扩增,DNA测序正确后,转化到大肠杆菌BL21（DE3）中,扩增后IPTG诱导表达。通过SDS-PAEG及Western-blot方法鉴定表达的融合蛋白。结果 成功构建人IL-15表达载体,并表达于大肠杆菌中。SDS—PAEG及Western-blot分析显示表达的融合蛋白分子量为16．1ku,与理论值相符。结论 获得了hIL-15融合蛋白,为hIL-15单克隆抗体的制备及其功能研究奠定了基础。     

STK15在头颈鳞状细胞癌组织中的表达及其对Hep-2细胞株生长的影响

目的: 探讨丝氨酸/苏氨酸激酶 15(STK15)在头颈鳞状细胞癌(鳞癌)组织中的表达及其与各临床病理特征的关系,阐明沉默STK15对人喉癌Hep-2细胞株生物学的影响。方法: 采用免疫组织化学方法检测头颈鳞癌组织中STK15蛋白表达;采用慢病毒沉默STK15,获得稳定转染细胞株;实验分为对照组(PLKO.1-puro-siRNA)和实验组(PLKO.1-puro-siSTK15),采用Western blotting法、RT-PCR 法、MTT法、侵袭实验和免疫荧光法检测沉默STK15对Hep-2细胞中STK15蛋白和STK15 mRNA表达水平、细胞增殖活性、穿膜细胞百分比和中心体数量的影响。结果: 免疫组织化学检测,STK15蛋白在头颈鳞癌组织中的表达率为64.96%,STK15蛋白表达与头颈鳞癌的TNM分期和预后有密切关联(P<0.05)。Western blotting法检测,实验组STK15蛋白表达水平明显降低(P<0.05)。RT-PCR检测,实验组STK15 mRNA表达水平明显降低(P<0.05)。MTT法检测,实验组细胞增殖活性明显低于对照组 (P<0.05)。侵袭实验,实验组穿膜细胞百分比明显低于对照组 (P<0.05)。免疫荧光检测,实验组中心体数目明显低于对照组 (P<0.05)。结论: STK15蛋白表达与头颈鳞癌的发展和预后密切关联,沉默STK15可降低Hep-2细胞的生长速度和迁移能力,提示STK15可作为头颈鳞癌的治疗靶点和预后标志物。     

12/15-脂氧化酶基因敲除对肥胖相关性肾小球疾病模型小鼠肾组织的保护作用及其机制

探讨12/15-脂氧化酶（12/15-LO）基因敲除（12/15-LOKO）对高脂饮食（HFD）诱导的肥胖小鼠肾组织的保护作用，阐明其在肾脏疾病进展中的作用。     

日本血吸虫卵对TNBS诱导小鼠结肠炎肠黏膜表达NOD2/CARD15的影响

 目的 研究日本血吸虫卵对2,4,6-三硝基苯磺酸（2,4,6-trinitrobenzesulfonic acid,2,4,6-TNBS）诱导小鼠结肠炎肠黏膜表达NOD2/CARD15的影响。方法 实验小鼠（n=50）随机分成3组:正常对照组（n=10）、TNBS+生理盐水组（n=20）和TNBS+日本血吸虫卵组（n=20）,后两组用TNBS溶液灌肠（100 mg/kg）建立结肠炎模型,TNBS+日本血吸虫卵组在造模前第14天和第3天分别给予腹腔注射冰冻灭活血吸虫卵10 000个（1 mL冰生理盐水混悬液）,TNBS+生理盐水组同时给予相同体积的冰生理盐水腹腔注射,建模后第7天处死存活小鼠,用荧光定量RT-PCR（Real time PCR）法测定结肠组织的NOD2的mRNA基因表达,Western blot方法测定结肠组织NOD2蛋白表达水平。结果 TNBS+日本血吸虫卵组死亡率明显下降,结肠肉眼及组织病理炎症程度明显减轻;荧光定量RT-PCR分析显示,TNBS+生理盐水组较正常组结肠黏膜NOD2 mRNA相对表达量显著增加（P<0.01）,TNBS+日本血吸虫卵组较TNBS+生理盐水组NOD2 mRNA相对表达量显著下降（P<0.05）;Western blot分析显示,TNBS+生理盐水组NOD2的蛋白表达量较正常组增加了近3倍（P<0.01）,TNBS+日本血吸虫卵组较TNSB+生理盐水组下降52.8%（P<0.01）,差异有显著统计学意义。结论 结肠炎时,黏膜NOD2/CARD15表达明显升高,日本血吸虫卵抗原可能通过下调NOD2/CARD15表达改善结肠炎症状态。     

应用CRISPR/Cas9技术构建IL-15敲除的MGC-803胃癌细胞株

目的:运用CRISPR/Cas9基因编辑技术,建立IL-15基因敲除的MGC-803胃癌细胞株?方法:针对IL-15基因作用的功能域,设计了靶向IL-15基因Exon2的gRNA? 构建PX458-gRNA重组质粒并转化感受态细胞Stbl3,筛选出重组子后进行测序,通过测序确认了所设计的gRNA的有效性?并进一步通过流式细胞分选以及ELISA法检测筛选出的MGC-803胃癌细胞株IL-15基因的表达水平?结果:测序结果显示敲除质粒构建成功,与阴性对照组相比,转染PX458-IL-15-gRNA质粒组IL-15的表达水平明显低于阴性对照组,差异具有统计学意义(P < 0.001)?结论:利用CRISPR-Cas9系统成功构建了IL-15基因敲除的MGC-803 细胞,为后续研究IL-15在肿瘤中的作用机制和功能奠定了基础?     

15FQ+量表中文版信度与效度分析

目的: 探讨个性因素量表(15FQ )中文修订版的信度和效度. 方法: 大学本科生1540名,分别进行相应量表的测量,包括15FQ 量表中、英文版以及中文效标量表16项人格因素问卷(16PF)、EPQ和MBTI,并对测量结果进行统计分析. 结果: ① 15FQ 量表中文版内部一致性系数为0.61～0.80,重测信度(相隔4 wk)为0.64～0.85(P<0.01);② 中英文版本各维度相关显著(0.42～0.88, P<0.01);③效标关联效度表明,中文版15FQ 量表各维度很好地表征了16项人格特质;④ 因子分析各题目最大负荷值落在主因素上占75%～100%,仅6题目因子分析不理想. 结论: 修订的15FQ 量表中文版具有较好的信度和效度,可以在中国进一步推广使用.     

p15基因在胆管癌中的表达及意义

目的研究p15在胆管癌中的表达并探讨其与临床分期、分级和淋巴结转移的关系及意义。方法应用超敏SP免疫组化方法检测38例胆管癌和8例胆总管囊肿中p15蛋白的表达情况。结果p15蛋白在胆管癌中的阳性表达率为52．63％,明显低于在胆总管囊肿中的表达率100％,有显著统计学差异（P〈0．05）;p15蛋白表达率在Ⅰ～Ⅲ级逐级下降,Ⅰ级的表达率高于Ⅲ级（P〈0．05）;Ⅰ～Ⅳ期表达率逐期下降,Ⅰ＋Ⅱ期的表达率高于Ⅲ＋Ⅳ期的表达率（P〈0．05）;有淋巴结转移癌组织中的阳性表达率为25．00％,显著低于在无淋巴结转移癌组织中的阳性率72．73％,（P〈0．05）。结论p15可能参与了胆管癌的发生、发展和转移,检测p15蛋白的表达对评估胆管癌的分期、分级及淋巴结转移有重要意义。