组蛋白去甲基化酶含十字形结构域蛋白-3和核因子-κB高表达与肝细胞癌不良进展和预后的关系

目的:观察肝细胞癌中组蛋白去甲基化酶含十字形结构域蛋白-3(JMJD3)和核因子-κB(NF-κB)蛋白水平的表达,探讨两者的相关性及其蛋白表达水平与多个临床病理因素的关系。方法:采用实时荧光定量反转录-聚合酶链反应(RT-qPCR)法检测25例新鲜肝细胞癌组织及相对应的癌旁组织中JMJD3和NF-κB信使RNA(mR...     

两用式胃管固定装置的研制与应用

目的研制两用式胃管固定装置,并探讨其使用效果。方法将132例留置胃管患者随机分为观察组和对照组各66例,两组均按照常规置入胃管,对照组用胶布固定,观察组采用自行研制的两用式胃管固定装置固定胃管。结果两组胃管置入时间及胃管脱出率、局部皮肤反应发生率比较,差异有统计学意义(均P0.01)。结论两用式胃管固定装置使用简便安全,固定牢固,适用于医院、社区及家庭长期留置胃管的患者。     

Hyperbaric oxygen treatment prevents nitric oxide‐induced apoptosis in articular cartilage injury via enhancement of the expression of heat shock protein 70

Heat shock proteins (HSPs), inflammatory cytokines, nitric oxide (NO), and localized hypoxia‐induced apoptosis are thought to be correlated to the degree of cartilage injury. We investigated the effect of hyperbaric oxygen (HBO) on (1) interleukin‐1β (IL‐1β)‐induced NO production and apoptosis of rabbit chondrocytes and (2) healing of articular cartilage defects. For the in vitro study, RT‐PCR and Western blotting were performed to detect mRNA and protein expressions of HSP70, inducible NO synthase (iNOS), and caspase 3 in IL‐1β‐treated chondrocytes. To clarify that the HSP70 was necessary for anti‐iNOS and anti‐apoptotic activity by HBO, we treated the cells with an HSP70 inhibitor, KNK437. For the in vivo study, cartilage defects were created in rabbits. The HBO group was exposed to 100% oxygen at 2.5 ATA for 1.5 h a day for 10 weeks. The control group was exposed to normal air. After sacrifice, specimen sections were sent for examination using a scoring system. Immunohistochemical analyses were performed to detect the expressions of iNOS, HSP70, and caspase 3. Our results suggested that HBO upregulated the mRNA and protein expressions of HSP70 and suppressed those of iNOS and caspase 3 in chondrocytes. KNK437 inhibited the HBO‐induced downregulation of iNOS and casapase 3 activities. The histological scores showed that HBO markedly enhanced cartilage repair. Immunohistostaining showed that HBO enhanced HSP70 expression and suppressed iNOS and caspase 3 expressions in chondrocytes. Accordingly, HBO treatment prevents NO‐induced apoptosis in articular cartilage injury via enhancement of the expression of heat shock protein 70. © 2012 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 31: 376–384, 2013     

Prognostic Impact of Cell Division Cycle Associated 2 Expression on Pancreatic Ductal Adenocarcinoma

Objective To examine the expression of cell division cycle associated 2 (CDCA 2) in pancreatic ductal adenocarcinoma (PDAC) and investigate its role in prognosis of PDAC patients. Methods This retrospective study included 155 PDAC patients who underwent surgical treatment and complete post-operative follow-up. Clinicopathologic data were collected through clinical database. Tissue microarray was constructed and immunohistochemistry was performed to detect CDCA2 expression in the PDAC tumor tissues and adjacent non-tumor tissues. Clinicopathological characteristics between high and low CDCA2 expression were compared. Correlation of CDCA2 expressions with patients’ survival was analyzed using Kaplan-Meier method and Cox regression analysis. Results Expression of CDCA2 in PDAC cells was significantly higher than that in adjacent non-tumor tissues (U=4056.5,P<0.001). Univariate analysis showed that CDCA2 expression [hazard ratio (HR)=1.574, 95% confidence interval (CI)=1.014-2.443,P=0.043] and node metastasis (HR=1.704, 95%CI=1.183-2.454, P=0.004) were significantly associated with prognosis. Cox regression analysis showed CDCA2 expression was not an independent prognostic risk factor (HR=1.418, 95%CI=0.897-2.242,P=0.135) for PDCA patients. Stratification survival analysis demonstrated CDCA2 expression as an independent prognostic risk factor in male patients (HR=2.554, 95%CI=1.446-4.511,P=0.003) or in non-perineural invasion patients (HR=2.290, 95%CI=1.146-4.577,P=0.012). Conclusions CDCA2 is highly expressed in PDAC tumor tissue. Although CDCA2 is not an independent prognostic risk factor for PDAC patients, it might be used to help predict prognosis of male or non-perineural invasion patients of PDAC.     

Secure and Efficient Two-Factor User Authentication Scheme with User Anonymity for Network Based E-Health Care Applications

Benefited from the development of network and communication technologies, E-health care systems and telemedicine have got the fast development. By using the E-health care systems, patient can enjoy the remote medical service provided by the medical server. Medical data are important privacy information for patient, so it is an important issue to ensure the secure of transmitted medical data through public network. Authentication scheme can thwart unauthorized users from accessing services via insecure network environments, so user authentication with privacy protection is an important mechanism for the security of E-health care systems. Recently, based on three factors (password, biometric and smart card), an user authentication scheme for E-health care systems was been proposed by Amin et al., and they claimed that their scheme can withstand most of common attacks. Unfortunate, we find that their scheme cannot achieve the untraceability feature of the patient. Besides, their scheme lacks a password check mechanism such that it is inefficient to find the unauthorized login by the mistake of input a wrong password. Due to the same reason, their scheme is vulnerable to Denial of Service (DoS) attack if the patient updates the password mistakenly by using a wrong password. In order improve the security level of authentication scheme for E-health care application, a robust user authentication scheme with privacy protection is proposed for E-health care systems. Then, security prove of our scheme are analysed. Security and performance analyses show that our scheme is more powerful and secure for E-health care systems when compared with other related schemes.     

基于同源重组和反向PCR扩增技术构建BRD7溴区结构域缺失突变体

目的：利用同源重组和反向PCR扩增技术构建溴区包含蛋白7 (bromodomain-containing protein 7,BRD7)的溴区结构域(bromodomain)缺失的真核表达载体。方法：设计一对反向PCR引物,以预先构建好的pIRES2-EGFP-3Flag/BRD7质粒为模板,利用高保真酶的高保真性和较长的延伸能力,反向扩增出BRD7溴区结构域缺失突变体(pIRES2-EGFP-3Flag/BRD7△brd)的线性DNA片段,然后转化大肠杆菌,利用大肠杆菌同源重组修复能力即可自身环化,而不需要经连接酶连接或其他处理即可得到pIRES2-EGFP-3Flag/BRD7△brd重组表达质粒,通过测序和Western印迹进一步对该缺失突变体的序列和蛋白质表达性能进行验证。结果：通过酶切鉴定、测序和Western印迹证实pIRES2-EGFP-3Flag/BRD7△brd构建成功。结论：利用PCR反向扩增和同源重组技术,可以简便快速地构建pIRES2-EGFP-3Flag/BRD7△brd突变体,实验成本较低,并且发现质粒模板浓度会影响载体的同源重组效率,这种改进的技术可广泛应用到基因突变体的构建。     

CXCR4及NDRG1蛋白在前列腺癌中的表达及其与侵袭转移的相关性分析

目的 探讨趋化因子受体4(chemokine receptor 4, CXCR4)及N-myc下游调节基因1(N-myc downstream regulated gene 1, NDRG1)在前列腺癌组织及细胞系中的表达情况,明确其与前列腺癌恶性程度及转移的关系。 方法 应用免疫组织化学方法检测CXCR4蛋白在前列腺癌(46例)、良性前列腺增生(15例)及正常前列腺组织(10例)中的表达,分析其与前列腺癌病理分级及转移的关系。免疫印迹(Western blotting)法检测不同前列腺癌细胞系中CXCR4及磷酸化NDRG1(pNDRG1)表达差异。结果 免疫组织化学检测发现CXCR4蛋白在前列腺癌组织(30例)中过度表达,表达率为65.2%,其表达水平与前列腺癌病理分级及Gleason评分无显著相关性(P=0.081),而CXCR4蛋白表达与肿瘤转移密切相关,前列腺癌转移患者表达率明显高于无转移患者,差异有统计学意义(P=0.002)。Western blotting法检测在正常前列腺上皮细胞和不同前列腺癌细胞系中CXCR4及NDRG1蛋白表达不同,高转移潜能的去势抵抗前列腺癌细胞(PC3和DU145)中CXCR4表达较高,而作为转移抑制因子的pNDRG1表达则明显降低。结论 趋化因子受体CXCR4及转移抑制因子NDRG1可能参与调控前列腺癌的进展和转移,对前列腺癌的临床诊断和治疗有重要价值。     

血清HE4、CA125、VEGF联合检测对卵巢癌诊断价值分析

目的 探讨血清人附睾蛋白4（HE4）、癌抗原-125（CA125）、血管表皮生长因子（VEGF）联合检测对卵巢癌诊断价值,为临床诊断提供一定的参考价值。方法 本研究病例来源于2013年5月~2015年5月期间笔者医院收治的107例卵巢疾病患者,按照术后病理结果将其分为卵巢癌组43例与良性病变组64例。选择同期于笔者医院体检中心体检的健康女性50例作为健康对照组。检测各组血清HE4、CA125、VEGF水平变化及联合检测敏感度和特异性。结果 卵巢癌组和卵巢良性病变组血清HE4水平明显高于健康对照组（P<0.05）;卵巢癌组血清HE4水平明显高于卵巢良性病变组（P<0.05）;卵巢癌组和卵巢良性病变组血清CA125水平明显高于健康对照组（P<0.05）;卵巢癌组血清CA125水平明显高于卵巢良性病变组（P<0.05）;卵巢癌组和卵巢良性病变组血清VEGF水平明显高于健康对照组（P<0.05）;卵巢癌组血清VEGF水平明显高于卵巢良性病变组（P<0.05）;血清HE4+CA125+VEGF联合检测敏感度和特异性明显高于单检测血清HE4、CA125、VEGF（P<0.05）。结论 血清HE4、CA125、VEGF在卵巢癌患者中呈高表达,3种肿瘤标志物联合检测具有着较高的敏感度和特异性,可作为诊断卵巢癌的重要手段,具有重要研究价值。     

A simple automated stimulator of mechanically induced arrhythmias in the isolated rat heart

Transient stretching of the ventricle can trigger arrhythmias and evoke ventricular fibrillation, especially when the stimulation occurs in the vulnerable period. To explore the sensitivity of small hearts we used a commercial pressure servo to study the kinetic relationship of left ventricular pressure to excitability and arrhythmias in the rat heart. Stimulation protocols were readily composed on the computer and programmed to vary the stimulus amplitude and timing relative to pacing. The pressure-induced premature ventricular excitations were similar to those observed in larger hearts, but the convenience of using small hearts allows the use of inexpensive transgenic animals to explore the molecular basis of transduction.