稳定表达人α-分泌酶adam10基因启动子荧光素酶报告基因细胞系的构建研究

目的 构建携带adam10基因启动子的荧光素酶报告载体,筛选稳定表达细胞系并分析其活性.方法 提取人神经母细胞瘤细胞(SH-SY5Y细胞)基因组DNA,以其为模板,PCR扩增adam10基因启动子并克隆至荧光素酶报告载体pGL4.17中,构建adam10基因启动子荧光素酶报告载体pGL4.17-adam10,将其转染SH-SY5Y细胞(无启动子的pGL4.17载体作阴性对照,带有CMV启动子的pGL4.51载体作阳性对照),经G418进行稳定表达株的筛选,用1μmol/L维甲酸处理细胞4d后检测其荧光活性.结果 成功扩增到438 bp的adam10基因启动子,pGL4.17-adam10经PCR和双酶切鉴定均正确.SH-SY5Y细胞被该载体转染后经G418筛选得到稳定表达adam10基因启动子的细胞株,经检测具有较强的转录活性;1μmol/L雏甲酸能诱导adam10基因启动子高效表达.结论 成功构建了人adam10基因启动子荧光素酶报告载体,adam10基因启动子在SH-SY5Y细胞中能稳定表达,为深入研究adam10基因的表达调控、多态性分析及其高通量药物筛选提供基础.     

甲基莲心碱对家兔、大鼠及人血小板聚集功能的影响

以阿斯匹林为对照,观察甲基莲心碱(Nef)对家兔、大鼠及人血小板聚集功能的影响。结果表明,Nef体外能明显抑制ADP、Adr、胶原诱导的不同种属动物的血小板聚集,呈剂量依赖性。对ADP、胶原诱导的家兔血小板聚集IC_(50)分别为16、22 μmol/L;对ADP、胶原、Adr诱导的人血小板聚集的IC_(50)分别为35、102、89 μmol/L。Nef 500 μmol/L明显降低ADP诱导的高脂喂养大鼠的血小板聚集率,并出现解聚。等摩尔浓度时,Nef作用强于阿斯匹林。     

气郁质与失眠相关性的病例对照研究

目的：通过对睡眠质量以及中医体质的调查探索失眠与气郁质之间的关系。 方法：对168名16~80岁的志愿者用中医体质量表、匹兹堡睡眠质量量表（Pittsburgh Sleep Quality Index, PSQI）、爱波沃斯嗜睡量表（Epworth Sleepiness Scale, ESS）以及深睡眠量表（Deep Sleep Scale, DSS）进行问卷调查。采用病例对照的方法分析气郁质和其他体质的睡眠质量和主要失眠症状。 结果：分析结果显示气郁质患者睡眠障碍的症状主要表现为入睡困难（PSQI： OR=3.012, 95% CI=1.310~6.923; DSS： OR=3.016, 95% CI=1.358~6.709）以及早醒（PSQI： OR=3.545, 95% CI=1.229~10.232; DSS： OR=2.742, 95% CI=1.072~7.014）;而其他体质的人群主要表现为多梦（PSQI： OR=2.419, 95% CI=1.154~5.072; DSS： OR=2.561, 95% CI=1.116~5.880）。失眠症状与气郁质的程度之间存在相关关系,分析结果显示气郁质越严重,其发生入睡困难、早醒的危险系数越高。 结论：本研究结果显示气郁质与失眠之间存在显著的相关性,对失眠的治疗与康复可以从调节中医体质的角度做进一步的研究。     

MRI软骨下骨髓水肿与早期膝骨关节炎临床表现的相关性研究?

目的：探讨早期膝骨关节炎(KOA)MRI 软骨下骨髓水肿的程度,并探究骨髓水肿与 KOA 夜间休息痛、关节肿胀程度和骨关节炎功能(the western Ontario and McMaster Universities osteoarthritis index,WOMAC)评分的相关性。方法本研究按纳排标准收集了早期 KOA 病例60例,其中伴骨髓水肿改变30例,设为观察组,不伴骨髓水肿改变30例,设为对照组。观察骨髓水肿程度评分、夜间休息痛视觉模拟(visular analogue scale, VAS)评分、关节肿胀程度及 WOMAC 功能评分情况。结果1)两组患者在夜间休息痛 VAS 评分、关节肿胀程度和 WOMAC 评分上比较,差异有统计学意义(P <0.05);2)观察组骨髓水肿程度越重,夜间休息痛 VAS 评分与WOMAC 评分越高;3)观察组骨髓水肿和夜间休息痛情况的相关系数 r =0.933(r >0)(P <0.05),二者呈正相关性;骨髓水肿与 WOMAC 评分的相关系数 r=0.555(r>0)(P <0.05),二者呈正相关性;骨髓水肿同关节肿胀程度相关系数 r=0.148(r>0)(P >0.05),二者间无明显相关性。结论MRI 软骨下骨髓水肿样改变与早期 KOA 夜间休息痛、WOMAC 评分具有正相关性;与关节肿胀情况无相关性。     

Biomarkers of endothelial dysfunction and risk of type 2 diabetes mellitus

Context  Endothelial dysfunction occurs in diagnosed type 2 diabetes mellitus but may also precede development of diabetes. Objective  To determine whether elevated plasma levels of biomarkers reflecting endothelial dysfunction (E-selectin; intercellular adhesion molecule 1 [ICAM-1]; and vascular cell adhesion molecule 1 [VCAM-1]) predict development of type 2 diabetes in initially nondiabetic women. Design and Setting  Prospective, nested case-control study within the Nurses' Health Study, an ongoing US study initiated in 1976. Participants  Of 121 700 women initially enrolled, 32 826 provided blood samples in 1989-1990; of those free of diabetes, cardiovascular disease, or cancer at baseline, 737 developed incident diabetes by 2000. Controls (n = 785) were selected according to matched age, fasting status, and race. Main Outcome Measure  Risk of confirmed clinically diagnosed type 2 diabetes by baseline levels of E-selectin, ICAM-1, and VCAM-1. Results  Baseline median levels of the biomarkers were significantly higher among cases than among controls (E-selectin, 61.2 vs 45.4 ng/mL; ICAM-1, 264.9 vs 247.0 ng/mL; VCAM-1, 545.4 vs 526.0 ng/mL [all P values .004]). Elevated E-selectin and ICAM-1 levels predicted incident diabetes in logistic regression models conditioned on matching criteria and adjusted for body mass index (BMI), family history of diabetes, smoking, diet score, alcohol intake, activity index, and postmenopausal hormone use. The adjusted relative risks for incident diabetes in the top quintile vs the bottom quintiles were 5.43 for E-selectin (95% confidence interval [CI], 3.47-8.50), 3.56 for ICAM-1 (95% CI, 2.28-5.58), and 1.12 for VCAM-1 (95% CI, 0.76-1.66). Adjustment for waist circumference instead of BMI or further adjustment for baseline levels of C-reactive protein, fasting insulin, and hemoglobin A_1c or exclusion of cases diagnosed during the first 4 years of follow-up did not alter these associations. Conclusion  Endothelial dysfunction predicts type 2 diabetes in women independent of other known risk factors, including obesity and subclinical inflammation.      

重庆市51家医院手术患者安全核查现状调查

目的了解重庆市手术患者安全核查现状,以确保手术患者的安全。方法采用自行设计的问卷,对2012年9月参加重庆医科大学附属第一医院主办的国家级继续教育项目培训班的104名护士进行调查。结果手术安全核查表实施率、住院患者应用手术安全核查表均为100%,二级医院和三级医院参与核对的人员按规范执行的分别占82.86%、87.50%,采用手术部位标记识别的分别占77.14%、93.75%,有定期检查督促的分别占82.86%、75%,有定期培训的分别占82.86%、87.50%,患者身份辨别采用佩戴腕部标志、预防性抗菌药物使用地点这两项,三级医院优于二级医院,差异有统计学意义(P<0.05)。结论重庆市51家医院手术患者安全核查全部落实,不同级别医院手术患者安全核查的具体做法存在差异,建议各级医院进一步加强手术患者安全核查的监管与落实,以确保手术患者的安全。     

食管鳞癌组织中HIF-1α和EGFR蛋白的表达与预后的关系

目的研究食管鳞状细胞癌(ESCC)中HIF-1α、EGFR蛋白的表达与食管癌病理特征及预后的关系。方法采用免疫组织化学(SP)法检测116例ESCC组织中HIF-1α、EGFR蛋白的表达,并分析其与临床病理因素及患者生存的关系。结果 HIF-1α与EGFR在ESCC组织中的阳性率分别为63.8%和68.1%,HIF-1α与EGFR均阳性率为50.9%。HIF-1α蛋白的阳性表达与T分期相关(P=0.014)。联合检测HIF-1α和EGFR蛋白均阳性表达与浸润深度及临床分期相关(P=0.006,0.023)。HIF-1α与EGFR蛋白表达呈正相关(r=0.331,P=0.000)。Kaplan-Meier生存分析结果显示,EGFR蛋白阳性表达的ESCC患者生存时间短(P=0.019)。结论 HIF-1α、EGFR蛋白在ESCC组织中呈高表达,二者密切相关,在ESCC发生发展中有重要作用,EGFR可作为判断预后的重要指标。     

Highly sensitive and convenient aptasensor based on Au NPs@Ce-TpBpy COF for quantitative determination of zearalenone

In this work, an aptasensor based on a portable U-disk electrochemical workstation in combination with a screen-printed electrode (SPE) is demonstrated for the quantitative determination of zearalenone (ZEN). The aptamer is immobilized on Au NPs@Ce-TpBpy COF (Covalent organic frameworks), which is modified on the surface of glassy carbon electrode. ZEN specifically binds to ZEN aptamer, which hinders the electron transfer and decreases the catalytic current of Au NPs@Ce-TpBpy COF for the reduction of hydrogen peroxide, measured by chronoamperometry (i–t). The quantitative detection of ZEN toxin is realized by a decrease of the catalytic current (ΔI). Under the optimal experimental conditions, the aptamer sensor exhibited excellent sensitivity, selectivity, reproducibility. A wide linear range of 1 pg mL⁻¹–10.0 ng mL⁻¹ with a detection limit of 0.389 pg mL⁻¹ (at 3σ) was obtained. The linear equation is ΔI = 0.401 lg c + 1.948 with a correlation coefficient of 0.9906. The recovery is in the range of 93.0–104.7% for the cornflour samples. The proposed method offers a new strategy for the rapid, inexpensive, and real-time detection of ZEN.

An aptasensor based on a portable U-disk electrochemical workstation is demonstrated for the quantitative determination of zearalenone. The aptamer sensor exhibited excellent sensitivity, selectivity, reproducibility.     

新型冠状病毒肺炎后潜水员适潜性评估

新型冠状病毒肺炎（COVID?19）自2019年底爆发以来,对人类生命健康造成了极其严重的危害。作为一种呼吸系统疾病,COVID?19对全身几乎所有器官都会产生影响,其中最主要的受累器官为肺和心脏。随着COVID?19 “康复”患者数量不断增加,越来越多的证据表明,因COVID?19而住院治疗得到“康复”的患者中有相当大比例的人群存在着后遗症,主要表现为呼吸系统和心血管系统的症状。对于潜水员,COVID-19后运动耐受、心肺功能的恢复对于其适潜性具有重要的影响。一些组织和机构对于潜水员COVID-19后的适潜性评估,发布了指南。本文对COVID-19后呼吸和心血管后遗症以及相关的适潜性评估方案进行简要介绍,并基于已有的相关要求和我国实际,初步提出我国潜水员COVID-19后的适潜性评估要求。     

重度二尖瓣狭窄合并小左心室的外科治疗分析

目的总结重度二尖瓣狭窄合并小左心室的手术治疗经验。方法回顾性分析行二尖瓣置换术（MVR）的115例重度二尖瓣狭窄患者的临床资料。根据是否合并小左心室将患者分为小左心室组（61例）和非小左心室组（54例）,对两组患者术后早期并发症发生率及病死率进行比较。结果小左心室组术后早期病死率、并发症发生率明显高于非小左心室组[14．75％（9／61）比7．41％（4／54）、55．74％（34／61）比25．93％（14／54）],差异均有统计学意义（P〈0．05）。结论积极而有效的围手术期处理及正确的术中决策是降低重度二尖瓣狭窄合并小左心室患者MVR术后早期并发症发生率及病死率的关键。