pLKO.1-SOCS3-siRNA慢病毒表达载体的构建与鉴定

目的:构建人类SOCS3的慢病毒表达载体pLKO.1-SOCS3-siRNA.方法:设计以SOCS3为靶标的siRNA并克隆进入pLKO.1载体,用基因测序进行重组克隆验证.结果:PCR检测证实siRNA插入pLKO.1质粒,测序分析证实插入序列正确.结论:成功构建SOCS3基因的siRNA载体,为研究SOCS3基因对宫颈癌细胞增殖、凋亡的影响提供了稳定转染的骨架载体.     

H19 DMR methylation correlates to the progression of esophageal squamous cell carcinoma through IGF2 imprinting pathway

Background

H19 gene has been proved to be essential for human tumor growth which contains CpG rich regions. Imprinted gene expression in many cancers is usually associated with the function of methylation. We performed this study to better understand wether H19 DMR methylation correlates to the progression of esophageal squamous cell carcinoma through IGF2 imprinting pathway.

Methods

LOI of IGF2 was detected in 276 samples, which were determined as heterozygote with ApaI polymorphism in exon 9 of IGF2 by PCR–RFLP and RT-PCR–RFLP. Methylation status of H19 DMR in informative samples was analyzed by bisulfite sequencing PCR. IGF2 expression was examined by real-time PCR and IHC.

Results

208 ESCC patients were informative for ApaI polymorphism. 92 tumor and 30 normal tissues showed IGF2 LOI. Methylation status of H19 CBS6 was higher in patients with IGF2 LOI compared to patients with IGF2 MOI (p < 0.05). IGF2 expression in patients with IGF2 LOI was higher than patients with IGF2 MOI (p < 0.05) which was correlated with lymph node involvement, neoplastic grade and metastasis (p < 0.05).

Conclusions

Our results suggested that H19 CBS6 hypermethylation is related to the LOI of IGF2 which usually leads to an overexpression of IGF2, playing important roles in the occurrence, development as well as metastasis of ESCC. Therefore, H19 CBS6 methylation potentially represents a novel clinically relevant epigenetic marker to identify individuals at increased risk for the occurrence, progression and prognosis of ESCC.     

Changes in circumpapillary retinal vessel density after acute primary angle closure episode via OCT angiography

Purpose

To investigate the changes and evaluate the diagnosis value of circumpapillary vessel density (VD) in cases of acute primary angle closure (APAC).

Design

Case–control study.

Methods

APAC patients with a history of unilateral acute attack were enrolled. The eyes with acute episode constituted the case group while the contralateral eyes without attack consisted of the control group. Ophthalmic examinations including slit-lamp examination, best-corrected visual acuity, intraocular pressure and visual field were carried out. Retinal nerve fiber layer (RNFL), macular ganglion cell complex (GCC) were measured by spectral-domain optical coherence tomography, while VD was assessed by optical coherence tomography angiography.

Results

The whole en face image vessel density (wiVD), circumpapillary vessel density (cpVD) and inside disk VD for both all vessels and capillary were all significantly lower in the APAC eyes compared to the fellow eyes (P?<?0.01 for all). In APAC eyes, the wiVD, inside disk VD and cpVD both for all vessels and capillary were all positively correlated with RNFL and GCC thicknesses but negatively correlated with the mean deviation (MD), pattern standard deviation (PSD) and the duration of acute attack (all P?<?0.01). From the ROC curve, the cpVD_cap, wiVD_cap, cpVD_all and wiVD_all all showed comparable diagnostic ability with RNFL, GCC and MD to differentiate eyes with APAC from the fellow eyes (all P?>?0.05). The inside disk VD_cap and VD_all demonstrated significant lower diagnostic ability than the cpVD_cap, wiVD_cap, cpVD_all and wiVD_all (all P?<?0.001).

Conclusions

In APAC eyes, circumpapillary VD decreased significantly compared with the fellow unaffected eyes. They were significantly correlated with thicknesses of RNFL and GCC, and visual field MD and PSD in the APAC eyes. The patients with longer duration of acute attack were more likely to have lower cpVD. For APAC, the diagnostic ability of wiVD and cpVD was similar with RNFL, GCC and MD and was higher than inside disk VD.

     

妇科腹腔镜手术的护理配合体会

目的：探讨妇科腹腔镜手术护理配合的经验方法.方法： 回顾性分析手术室收治的妇科腹腔镜手术患者的临床资料,进行总结和分析.结果： 妇科腹腔镜手术具有创伤小、痛苦轻、疗程短等优点,熟练地手术护理配合技巧能保证手术顺利进行,患者满意率高.结论： 妇科腹腔镜手术护理配合不仅能增加手术的安全性,并能减轻了医护工作强度.     

TNP-470联合5-FU抑制结肠癌生长的实验研究

背景与目的：新生血管生成是肿瘤生长转移的一个重要条件,本研究通过体内外实验观察血管生成抑制剂TNP-470联合5-氟尿嘧啶（5-fluorouracil,5-Fu）对结肠癌生长的影响。方法：采用四甲基偶氮唑盐（MTT）法检测TNP-470和5-Fu对LOVO细胞的生长抑制效应,用Burgi法分析联合用药效果。建立结肠癌皮下移植瘤模型,观察TNP-470单独或联合5-Fu对移植瘤的生长抑制作用,采用免疫组化和图像分析系统定量检测肿瘤组织中血管内皮生长因子（VEGF）的表达,抗Ⅷ因子抗体标记计数肿瘤微血管密度（MVD）。结果：TNP-470和5-Fu对体外培养的LOVO细胞的生长具有抑制作用,半数抑制浓度（IC50）分别为55.8ng/ml和4.6ng/ml,Burgi法分析表明两者联合应用具有协同效应,LOVO细胞生长进一步受到抑制。体内实验表明：各治疗组肿瘤的生长明显受到抑制,而联合治疗组抗瘤作用进一步增强。联合治疗组和5-Fu组肿瘤组织中VEGF的表达较对照组显著减少,联合治疗组和TNP-470组肿瘤组织中的MVD与对照组比较也显著降低。结论：TNP-470能够抑制LOVO细胞及其皮下移植瘤的生长和新生血管形成,与5-Fu联合应用具有协同作用,可提高抗瘤效应。     

Presence of active gelatinases in endometrial carcinoma and correlation of matrix metalloproteinase expression with increasing tumor grade and invasion

BACKGROUND: The actions of the extracellular-matrix degrading enzymes, matrix metalloproteinases (MMPs), are implicated in tumorigenesis. The cellular localization of MMP-2, MMP-9, membrane type 1 (MT1)-MMP, tissue inhibitors of metalloproteinases (TIMPs) 1-3, and the presence of active gelatinases were investigated in endometrial carcinoma. METHODS: Endometrial carcinomas were grouped according to histologic grade (Grades 1-3), depth of myometrial invasion (0, < 50%, > 50%) and the presence of vascular/lymphatic invasion. Twenty-nine endometrial carcinoma biopsies were investigated immunohistochemically to determine the tissue localization of MMP-2 (gelatinase A), MMP-9 (gelatinase B), MT1-MMP, and TIMPs 1-3. In situ hybridization was performed to localize MMP-2 and MMP-9 mRNA. The presence of active gelatinases was assessed using in situ zymography. RESULTS: Epithelial tumor cells were the main site of MMP-2, MMP-9, and MT1-MMP protein. Variable stromal cell localization was also observed, particularly in areas adjacent to tumor nests. Semiquantitative analysis revealed increases in MMP-9 and MMP-2 but not MT1-MMP staining scores in tumor epithelial cells in the transition from histologic Grade 1 to Grades 2 and 3. Matrix metalloproteinase-9 and MT1-MMP staining scores in tumor cells were significantly associated with the presence of myometrial invasion and vascular/lymphatic invasion, while MMP-2 did not correlate with these factors. In addition, MT1-MMP was co-localized with MMP-2, supporting its role in the activation of proMMP-2. Tumor cells from all histologic grades stained intensely for TIMP-2 and TIMP-3 proteins, while variable stromal staining was observed. In Grade 1 carcinomas TIMP-1 was predominantly immunolocalized to the stromal compartment with variable tumor cell localization being observed in Grades 2 and 3 carcinomas. Matrix metalloproteinase-9 and MMP-2 mRNAs were predominantly observed in tumor epithelial cells as well as in the stroma to varying degrees. In situ zymography revealed active forms of gelatinases at the cellular surface and in association with tumor epithelial cells within endometrial carcinoma tissues. CONCLUSIONS: These data suggest that increasing expression of MMPs and endometrial carcinoma progression are closely related. Active gelatinases are present in endometrial carcinoma, resulting in alterations to the microenvironment that promote tumor invasion and metastasis.     

临床常用氨基糖苷在烫伤兔体中的药动学改变

１７只家兔烫伤前和烫伤后７天静脉注射庆大霉素３ｍｇ／ｋｇ（ｎ＝７）和丁胺卡那霉素１０ｍｇ／ｋｇ（ｎ＝１０）的药动学资料比较的结果表明，２０％体表面积三度烫伤后家兔的庆大霉素和丁胺卡那霉素血浓度均明显下降；两药分布容积（Ｖ＿ｄ）均增加，半衰期（ｔ＿（１／２））缩短和药时曲线下面积（ＡＵＣ）减小。本实验证实，烫伤能改变庆大霉素和丁胺卡那霉素的药动学井导致伤后两药血浓度降低。     

一例输入性外籍伤寒病例的流行病学调查分析

目的分析1例国外输入性伤寒病例流行病学特征。方法对1例来自印度的输入性外籍发热病例进行流行病学调查,总结流行病学、临床和实验室检查结果,分析其特点。结果通过认真细致的流行病学调查结合临床表现和实验室检查该患者被确诊为输入性不典型伤寒,临床治愈出院,期间未发生疫情扩散。结论对来自国外的输入性发热病例应采取严格的隔离防控措施,逐一排查病因,尽早确诊,规范治疗,有效防止疫情自境外传入和扩散。