Molecular magnetic resonance probe targeting VEGF165: preparation and in vitro and in vivo evaluation

A new method for imaging the tumor human vascular endothelial growth factor 165 (VEGF 165) is presented. A magnetic resonance imaging (MRI) probe was prepared by crosslinking ultrasmall superparamagnetic iron oxide (USPIO) nanoparticles to the aptamer for tumor vascular endothelial growth factor 165 (VEGF165‐aptamer). The molecular probe was evaluated for its in vitro and in vivo activities toward VEGF165. Enzyme‐linked immunosorbent assay showed that the VEGF165‐aptamer–USPIO nanoparticles conjugate specifically binds to VEGF165 in vitro. A cell proliferation test showed that VEGF165‐aptamer–USPIO seems to block the proliferation of human umbilical vein endothelial cells induced by free VEGF165, suggesting that VEGF165 is an effective target of this molecular probe. In xenograft mice carrying liver cancer that expresses VEGF165, T₂‐weighted imaging of the tumor displayed marked negative enhancement 3 h after the intravenous administration of VEGF165‐aptamer–USPIO. The enhancement disappeared 6 h after administration of the probe. These results suggest the targeted imaging effect of VEGF165‐aptamer–USPIO probe in vivo for VEGF165‐expressing tumors. This is the first report of a targeted MRI molecular probe based on USPIO and VEGF165‐aptamer. Copyright © 2014 John Wiley & Sons, Ltd.     

人乳头状瘤病毒基因分型联合薄层液基细胞学检测在宫颈病变筛查中的应用

目的：探讨人乳头状瘤病毒（HPV）基因分型联合薄层液基细胞学（TCT ）检测在宫颈病变筛查中的应用价值。方法采用导流杂交基因芯片法检测473例宫颈疾病患者 HPV 基因型别,TCT 做细胞学筛查,同时行阴道镜活检,以组织学为金标准,对结果进行分析。结果473例患者共检出 HPV 阳性169例,总感染率为35．7％;473例患者高危型 HPV （HR‐HPV）感染率为32．1％,TCT 检测阳性率为26．6％,对宫颈病变筛查的灵敏度、特异度、漏诊率、阳性预测值和阴性预测值差异无统计学意义（χ2＝3．444,P＝0．063）;HR‐HPV 与 TCT 联合检测对宫颈病变筛查的灵敏度、特异度、漏诊率、阳性预测值、阴性预测值分别为95．8％、77．7％、4．2％、52．3％和98．7％,灵敏度和阴性预测值均较单独 HR‐HPV 及 TCT 检测明显提高,漏诊率明显降低,差异有统计学意义（P＜0．05）。结论 HPV 基因分型联合 TCT 检测能明显提高宫颈病变筛查的灵敏度和阴性预测值,降低漏诊率,更加有效地筛查宫颈病变。     

芍药苷对骨髓间充质干细胞增殖的影响

背景：研究显示芍药苷具有补血及治疗自身免疫性疾病的功效,骨髓间充质干细胞对机体的造血及免疫功能也起着重要的作用,但芍药苷对骨髓间充质干细胞的增殖及细胞因子的分泌和表达有何影响报道较少。目的：探讨芍药苷对人骨髓间充质干细胞增殖及白细胞介素6表达的影响。方法：采用密度梯度离心法和贴壁培养法体外分离培养人骨髓间充质干细胞,用流式细胞术和成脂及成骨诱导法鉴定人骨髓间充质干细胞生物学特性, MTT法检测不同浓度芍药苷对人骨髓间充质干细胞增殖的影响, EL ISA 法测定芍药苷干预人骨髓间充质干细胞后培养上清液中白细胞介素6的分泌水平,RT-PCR 检测芍药苷干预后白细胞介素6 mRNA的表达情况。结果与结论：成功分离出骨髓间充质干细胞,具有成骨、成脂分化潜能。与对照组相比,芍药苷浓度为2μmol/L和10μmo l/L可明显促进骨髓间充质干细胞增殖。10μmol/L 芍药苷干预骨髓间充质干细胞后, G0/G1期细胞比例显著降低, S期细胞比例显著升高。10μmol/L 芍药苷干预组骨髓间充质干细胞白细胞介素6 的分泌和mRNA表达均显著高于对照组(P <0.01)。由此得出,一定浓度的芍药苷可促进骨髓间充质干细胞增殖,并提高骨髓间充质干细胞分泌白细胞介素6水平和基因表达。     

免疫印迹法测定Tp47蛋白与梅毒患者血清的免疫反应性

目的克隆Tp47基因、构建原核表达载体、表达并纯化Tp47蛋白,通过免疫印迹法(Western-Blot)检测其免疫反应活性。方法聚合酶链反应扩增Tp47基因,将Tp47克隆至pGEX-6P-1载体,构建重组表达质粒pGEX-6P1-Tp47,经测序鉴定正确后,转化大肠埃希菌BL21(DE3),经诱导表达后,用亲和层析方法纯化重组蛋白,鉴定正确后采用Western-Blot检测重组蛋白与梅毒阴性、阳性血清的免疫反应性。结果成功构建pGEX-6P1-Tp47原核表达质粒,表达、纯化后获得了相对分子质量约为71×103的重组蛋白;Western-Blot检测显示其能与梅毒阳性患者血清发生特异性反应,而与梅毒阴性患者血清无交叉反应性。结论成功克隆、表达、纯化Tp47重组蛋白,其与临床各期梅毒患者血清具有较好的免疫反应性,为Tp47重组蛋白用于梅毒早期诊断提供了理论依据。     

不同体质量指数男性不育患者心理评估

目的：探讨不同体质量指数男性不育症患者的心理健康状况,为临床干预提供依据。方法将292例男性不育患者按照体质量指数分为3组,正常组84例,超重组117例,肥胖组91例。对3组患者应用症状自评量表进行测评分析。结果超重组和肥胖组症状自评量表总分及强迫、人际关系、抑郁、焦虑因子分均显著高于常模（P＜0．05）,总分及强迫因子分显著高于正常组（P＜0．05或0．01）;正常组总分及抑郁、焦虑因子分显著高于常模（P＜0．05）;入组被试体质量指数与强迫、抑郁、焦虑因子分呈显著正相关（P＜0．05）。结论肥胖和超重男性不育患者存在不同程度的心理问题,应对其予以有针对性的心理干预。     

游离雌三醇和人胎盘催乳素测定在子痫前期的意义

目的：探讨血清游离雌三醇(uE3)和人胎盘催乳素(hPL)与子痫前期的关系。方法选取2013年6月至2014年6月诊断为子痫前期的孕妇123例纳入观察组,同期健康孕妇150例纳入对照组,两组孕妇均分别在妊娠28~<32周和妊娠32~36周各采血一次,检测血清 uE3和 hPL 水平并进行比较。结果对照组妊娠结局优于观察组。随着孕周增加两组孕妇血清 uE3和 hPL 水平有升高趋势,在相同孕期中对照组血清 uE3和 hPL 水平均较观察组高,差异均有统计学意义(P <0.05)。结论子痫前期孕妇血清 uE3和 hPL 水平下降,与妊娠结局相关。     

血细胞计数影响因素分析

本文分析血细胞计数的影响因素,发现血细胞检测结果受环境温度、放置时间、采血方式等因素的影响,检验人员在采血和检测过程中应尽可能排除各种影响因素,为临床提供真实可靠的检验结果。     

动脉内皮细胞中TIE2启动CREG真核表达质粒pTIE2-CREG-EGFP-N1的表达

背景：血管内皮发生的分子机制是细胞及基因替代治疗的首要前提。目的：构建一种通过血管内皮细胞特异蛋白TIE2启动子来启动表达的E1A激活基因阻遏子（cellular repressor of E1A-stimulated genes,CREG）和增强绿色荧光蛋白（enhanced green fluorescence protein,EGFP）融合的真核表达质粒。方法：根据GenBank中公布的TIE2基因的启动子序列,人工合成TIE2启动子DNA序列,经AseⅠ和NheⅠ双酶切后,亚克隆入表达质粒pEGFP-N1中构建pTIE2-EGFP-N1。同时,用BamHⅠ和EcoRⅠ双酶切pcDNA3.1myc-His/hCREG质粒得到CREG基因,亚克隆入质粒pTIE2-EGFP-N1中构建pTIE2-CREG-EGFP-N1,酶切鉴定。应用脂质体法将该质粒转染至体外培养的小鼠动脉内皮细胞,荧光显微镜下观察EGFP的表达;Western blot检测CREG蛋白的表达。结果与结论：经酶切鉴定证实实验构建的pTIE2-CREG-EGFP-N1质粒正确;体外转染48h,荧光显微镜下可见EGFP的表达,Western blot检测到CREG蛋白的表达。说明实验成功构建了pTIE2-CREG-EGFP-N1重组质粒,其可携带目的基因在小鼠动脉内皮细胞中有效表达。     

The ability of current scoring systems in differentiating transient and persistent organ failure in patients with acute pancreatitis

Purpose

The purpose of this study is to investigate the accuracy of currently used scoring systems in differentiating transient and persistent organ failure in patients with acute pancreatitis (AP).

Materials and methods

In this retrospective study, 127 consecutive patients with AP and organ failure were included. Patients were divided into transient and persistent organ failure groups. The Acute Physiology and Chronic Health Examination II score, bedside index of severity in acute pancreatitis, harmless acute pancreatitis score, and modified Marshall scores within the first 24 hours of organ failure were collected, and their accuracy in predicting transient organ failure was assessed.

Results

Transient organ failure occurred in 46 patients (36.2%). Fewer patients with transient organ failure initiated with multiple organ failure (13.0% vs 37.0%, P = .004) and renal failure (17.4% vs 44.4%, P = .002). In predicting transient organ failure, the area under the curves of the 4 scoring systems is from 0.66 to 0.71. The area under the curve of serum amylase was 0.78, which was slightly better than that of the modified Marshall and Acute Physiology and Chronic Health Examination II score and was significantly better than that of the bedside index of severity in acute pancreatitis and harmless acute pancreatitis score (P < .05).

Conclusions

Current scoring systems are not accurate enough in differentiating transient and persistent organ failure in patients with AP.     

穴位注射骨髓间充质干细胞后心肌梗死大鼠血管细胞黏附因子1及迟发抗原4的表达

背景：目前认为骨髓间充质干细胞归巢是由黏附分子和趋化因子介导的,该过程有骨髓内皮细胞、造血干细胞、骨髓造血微环境及其分泌或表达的分子共同参与,黏附分子可能在其中起到重要作用。 目的：以血管细胞黏附因子1和迟发抗原4为指标探讨穴位注射骨髓间充质干细胞向心肌迁移、趋化的机制。方法：贴壁法培养骨髓间充质干细胞,以第3代为种子细胞,调整细胞浓度为1×10^10 L-1。60只SD大鼠随机分为假手术组、模型组、心肌注射骨髓间充质干细胞组（以下简称心肌组）,穴位注射骨髓间充质干细胞组（以下简称穴位组）,每组15只。采用结扎左冠状动脉前降支方法复制心肌梗死模型,穴位组造模成功72 h后于心俞、至阳、膻中每穴位注入骨髓间充质干细胞0.3 mL,心肌组造模成功72 h后二次开胸,左前降支供血区域及周边心肌分6点均匀地移植骨髓间充质干细胞1.2 mL,4周后颈动脉插管,多道生理记录仪检测血流动力学指标,ELISA法检测血清血管细胞黏附因子1及迟发抗原4表达水平。 结果与结论：心肌组、穴位组大鼠心功能得到改善,两组血清血管细胞黏附因子1及迟发抗原4较模型组升高,心肌组和穴位组比较差异无显著性意义。结果说明血管细胞黏附因子1/迟发抗原4轴可能是穴位注射干细胞趋化机制之一。