双向调控Cox-2表达对人食管癌EC9706细胞裸鼠移植瘤放射敏感性的影响

目的 探讨上调和下调Cox-2基因表达对食管癌EC9706细胞裸鼠移植瘤放射敏感性的影响。方法 构建针对Cox-2基因的siRNA载体及Cox-2基因真核表达载体,通过脂质体转染技术将其转入食管癌EC9706细胞,G418筛选得到稳定转染细胞系。荧光定量RT-PCR和Western blot分别检测细胞中Cox-2 mRNA和Cox-2蛋白表达水平;裸鼠移植瘤实验检测上调和下调Cox-2表达联合X射线照射对食管癌的生长抑制作用。结果 Cox-2下调组食管癌EC9706细胞内Cox-2基因表达明显降低,Cox-2上调组明显升高。Cox-2下调组裸鼠移植瘤平均体积明显小于对照组(F=34.26,P<0.05);Cox-2上调组的瘤体平均体积大于对照组(F=26.38,P<0.05)。20 Gy γ射线照射后Cox-2下调组瘤体平均体积较照射前明显减小(F=16.35,P<0.05);而上调组裸鼠皮下种植瘤平均体积较照射前无明显变化。结论 下调Cox-2表达能抑制人食管癌EC9706细胞裸鼠移植瘤的生长,增强瘤体的放射敏感性,上调Cox-2表达则使肿瘤辐射抗拒。     

对比剂注射部位对儿童复杂先天性心脏病双源CT成像的影响

目的探讨对比剂注射部位对儿童复杂先天性心脏病CTA图像质量的影响。方法回顾性分析426例复杂先天性心脏病患儿的双源CT心脏大血管CTA图像,其中经上肢静脉注射154例(A组),以下肢静脉注射272例(B组),对A、B组CTA图像上腔静脉对比剂硬化束伪影,心腔、大血管CT值,右心房、右心室对比剂均匀度,心内结构及大血管显示清晰度进行对比。结果 A组图像上腔静脉内对比剂硬化束伪影评分为3.00±1.04,B组为3.96±1.08,优于A组,二者差异有统计学意义(P0.01),两组心腔、大血管平均CT值的差异均无统计学意义(P均0.05);两组心内结构、大血管、冠状动脉起源及近段图像清晰度主观评分差异均有统计学意义,B组均高于A组(P均0.05),而右心房、右心室对比剂均匀度评分的差异无统计学意义(P均0.05)。结论对比剂注射部位的选择对儿童先天性心脏病CTA图像质量有显著影响,经下肢浅静脉注射对比剂有助于消除上腔静脉对比剂伪影,提高图像质量。     

Significance of Fas and FasL protein expression in cardiac carcinoma and local lymph node tissues

Objective: To investigate the relation of Fas and Fas ligand (FasL) protein expression with carcinogenesis and metastasis of cardiac carcinoma. Methods: Immunohistochemistry was used to detect Fas and FasL protein expression in 64 cardiac carcinoma tissue samples and 20 normal gastric tissue samples. Relation between FasL and Fas expression, age and gender of gastric cancer patients, and pathological subtype and lymph node metastasis of gastric cancer was analyzed. Results: The Fas expression level was significantly higher in normal gastric tissue samples than in cardiac carcinoma tissue samples (85.0% vs. 25.0%, P<0.001), while the FasL expression level was significantly lower in normal gastric tissue samples than in cardiac carcinoma tissue samples (30.0% vs. 81.3%, P<0.001). The Fas expression level was significantly higher in invasive lymph nodes than in non-invasive lymph nodes (82.9% vs. 56.5%, P<0.003) and in well-differentiated gastric carcinoma tissue samples than in poorly-differentiated cardiac carcinoma tissue samples (50.0% vs. 18.0%, P=0.015). The FasL expression level was significantly lower in well-differentiated cardiac carcinoma tissue samples than in poorly- differentiated cardiac carcinoma tissue samples (42.9% vs. 84.0%, P=0.021). The Fas and FasL expression levels (25.0% and 81.3%) were significantly different in cardiac carcinoma tissue samples (P<0.001), but had a non-linear correlation (P=0.575). Conclusion: Abnormal Fas and FasL expressions in cardiac carcinoma and lymph node tissues are involved in carcinogenesis and metastasis of gastric cancer.     

miR-126 regulated breast cancer cell invasion by targeting ADAM9

Accumulating evidence has shown that microRNAs (miRNAs) deregulation is commonly observed in human malignancies and crucial to cancer metastasis. Herein, we demonstrated that miR-126 play a suppressor role in human breast cancer cells invasion through the direct repression of a disintegrin and metalloprotease 9 (ADAM9). MiR-126 expression was investigated in forty cases of breast cancer specimens by real-time PCR. Transwell assay was conducted to explore the effects of miR-126 on the invasion of human breast cancer cell lines. The impact of miR-126 overexpression on putative target ADAM9 was subsequently confirmed by Western blot analysis. Our results indicated that miR-126 expression was frequently down-regulated in breast cancer specimens compared with adjacent normal tissues (P<0.05). Overexpression of miR-126 significantly reduced (P<0.05) the protein levels of ADAM9, further suppressed (P<0.05) breast cancer cell invasion in vitro. Meanwhile, knockdown of ADAM9 by small interfering RNA (siRNA) also inhibited (P<0.05) breast cancer cell invasion. Thus, our study revealed that miR-126 may act as a tumor suppressor via inhibition of cell invasion by downregulating ADAM9 in breast cancer development.     

Comparing triage algorithms using HPV DNA genotyping,HPV E7 mRNA detection and cytology in high-risk HPV DNA-positive women

BackgroundHigh-risk human papillomavirus (hrHPV) DNA positive women require triage testing to identify those with high-grade cervical intraepithelial neoplasia or cancer (≥CIN2).ObjectiveComparing three triage algorithms (1) E7 mRNA testing following HPV16/18/31/33/45/52/58 genotyping (E7 mRNA test), (2) HPV16/18 DNA genotyping and (3) cytology, for ≥CIN2 detection in hrHPV DNA-positive women.Study designhrHPV DNA-positive women aged 18–63 years visiting gynecology outpatient clinics were included in a prospective observational cohort study. From these women a cervical scrape and colposcopy-directed biopsies were obtained. Cervical scrapes were evaluated by cytology, HPV DNA genotyping by bead-based multiplex genotyping of GP5+6+-PCR-products, and presence of HPV16/18/31/33/45/52/58 E7 mRNA using nucleic acid sequence-based amplification (NASBA) in DNA positive women for respective HPV types. Sensitivities and specificities for ≥CIN2 were compared between E7 mRNA test and HPV16/18 DNA genotyping in the total group (n = 348), and E7 mRNA test and cytology in a subgroup of women referred for non-cervix-related gynecological complaints (n = 133).ResultsSensitivity for ≥CIN2 of the E7 mRNA test was slightly higher than that of HPV16/18 DNA genotyping (66.9% versus 60.9%; ratio 1.10, 95% CI: 1.0002–1.21), at similar specificity (54.8% versus 52.3%; ratio 1.05, 95% CI: 0.93–1.18). Neither sensitivity nor specificity of the E7 mRNA test differed significantly from that of cytology (sensitivity: 68.8% versus 75.0%; ratio 0.92, 95% CI: 0.72–1.17; specificity: 59.4% versus 65.3%; ratio 0.91, 95% CI: 0.75–1.10).ConclusionFor detection of ≥CIN2 in hrHPV DNA-positive women, an algorithm including E7 mRNA testing following HPV16/18/31/33/45/52/58 DNA genotyping performs similar to HPV16/18 DNA genotyping or cytology.     

多发性硬化实验动物模型研究进展

实验性自身免疫性脑脊髓炎(experimental autoimmune encephalomyelitis,EAE)是多发性硬化的实验动物模型,经过近2个多世纪的发展,已可以在猴、大鼠、小鼠等动物制备EAE模型,在同一种品系动物采用不同免疫抗原制备的EAE模型在敏感性和临床特点具有多样性。本研究就EAE的发展史和目前不同品系实验动物、不同抗原制备EAE模型的特点做一述评,对未来EAE模型的发展前景进行探讨。     

主动脉瓣关闭不全法建立新西兰兔心衰模型

目的 用主动脉瓣关闭不全法致新西兰兔心衰模型。方法 运用导管术造成新西兰兔主动脉瓣膜关闭不全, 制作超容量负荷型心衰模型。观察动物的毛色、精神状态、活动情况、饲料消耗指数、体重增加指数、呼吸频率等指标对该模型进行评价;检测血清SOD活力和MDA含量, 判断模型组动物机体抗氧化能力;利用酶联免疫法检测血清中cAMP、cGMP的变化情况;利用基因芯片技术分析该模型基因表达差异。结果 模型动物SBP、DBP和LVSP著性下降, LV+dp/dt和LV±dp/dt明显下降, LVDP显著上升。模型组与正常组比较, 毛发枯槁, 活动减少, 进食减少, 反应迟钝, 精神萎靡, 抓起时反抗减轻。模型组呼吸频率加快;模型组血清SOD活性低于正常组, MDA含量高于正常组;模型组血清cAMP明显低于正常组, cGMP明显高于正常组;利用基因芯片共检测出665个差异基因, 其中与心功能较密切相关的基因主要与离子通道、肌收缩、信号转导等功能有关。结论 采用主动脉瓣关闭不全法建立兔心衰模型方法可靠。主动脉关闭不全, 使心脏前负荷增加, 造成左心室舒张末期容积增加, 导致左心室扩大肥厚及心力衰竭。心肌组织基因芯片检测结果显示, 该模型基因表达出现了一定的变化。     

肥厚梗阻型心肌病介入治疗方法的新改进

目的 探讨经皮“室间隔心肌隧道化学消融术（percutaneous transluminal septal tunnel myocardial ablation,PTSTMA）”治疗传统技术不适合的肥厚梗阻型心肌病（hypertrophic obstructive cardiomyopathy,HOCM）的方法及疗效.方法 选择2005年6月至2011年6月期间住院的HOCM患者中的26例为研究对象.观察经PTSTMA治疗的26例HOCM患者术后即刻左心室流出道压力阶差（left ventricular outflow tract pressure gradient,LVOTPG）变化,术后24 h磷酸肌酸激酶同工酶、心电学改变,术后3个月心脏超声指标变化以及随访临床症状的转归.结果 3例通过单支血管消融,17例通过2支血管消融,6例通过3支血管消融.LVOTG由术前（75.6±22.4）mm Hg（1 mm Hg=0.133 kPa）降至（21.4±5.84）mm Hg,差异有统计学意义（P＜0.01）.术后24 h磷酸肌酸激酶同工酶为（1 86±84）μ/L,2例发生Ⅲ＆#176;房-室传导阻滞,均于1周后恢复正常传导,10例发生室性心律失常,12例发生右束支传导阻滞.消融后室间隔厚度减少[（16.8±4.2）mm vs.（22.8±5.8）mm,P＜0.01]、左心房内径减少[（42.0±8.6）mmvs.（48.0±7.0）mm,P＜0.05],差异均有统计学意义.随访时间为（39.8±8.6）个月.与消融前比较,随访中胸痛、呼吸困难症状明显减少,纽约心脏协会心功能分级明显改善,室性心律失常明显减少,黑蒙症状也有一定改善.结论 冠状动脉造影证实冠状动脉室间隔支解剖形态不适合做传统室间隔心肌化学消融术的HOCM患者,PTSTMA能显著降低LVOTPG,改善临床症状.PTSTMA可作为HOCM心肌化学消融术的一种补充方法,其近、中期安全有效.     

干燥综合征合并原发性胆汁性肝硬化患者的外周血Th17/Treg细胞百分比及临床特征

目的通过对SS合并PBC患者中外周血Th17和Treg细胞的检测,分析其临床特征,并探明Th17、Treg细胞对SS合并PBC患者的影响。方法选择2012年3月至2018年2月期间郑州人民医院收治的SS合并PBC患者22例,SS患者48例,选取健康志愿者68名。比较3组研究对象的肝功能指标、Th17、Treg和TH17/Treg水平。组间比较采用t检验或Kruskal-Wallis H检验;计数资料以百分比表示,组间比较采用卡方检验。结果SS合并PBC组出现黄疸、皮肤瘙痒、腹水、消化道出血、口干、食管静脉曲张、发热及眼部干涩等主要临床表现为20例(90.9%,20/22),高于对照组37例(77.1%,37/48),(P<0.05)。而体质量减轻的患者SS组(46例,95.8%)明显多于SS合并PBC组(17例,77.3%),(P<0.05)。SS合并PBC组患者ALT、AST、碱性磷酸酶(ALP)、γ-谷氨酰胺酶(γ-GT)和TBil均高于SS组和健康组(P<0.05)。SS合并PBC组患者Th17细胞百分比为[16.6(10.1,22.5)%],明显高于SS组[9.4(7.0,13.0)%,P<0.05]及健康组[4.9(4.3,9.8)%,P<0.05];SS合并PBC组Th17/Treg细胞比值[0.52(0.42,0.62)]均明显高于SS组[0.25(0.14,0.37),P<0.05]及健康组[0.22(0.13,0.26),P<0.05]。结论SS合并PBC患者外周血Th17/Treg细胞水平及其比值明显失衡,提示Th17细胞在很大程度上参与了致病过程。此外,Th17/Treg比值与某些肝功能指标有一定的相关性。