天津地区2013年百日咳感染状况及分子流行病学特征分析

目的 了解2013年天津地区百日咳感染状况及分子流行病学特点.方法 选取天津地区监测点181例百日咳疑似病例作为研究对象,采集鼻咽拭子和血清标本,应用Real-time PCR检测百日咳鲍特菌双目标基因,同时采用ELISA检测其特异性百日咳毒素IgG(PT-IgG).30份百日咳DNA阳性标本应用PCR扩增菌毛蛋白2(FIM2)、菌毛蛋白3(FIM3)基因,并对PCR产物进行DNA测序分析.结果 148例百日咳病例Real-time PCR检测阳性率为68.24%;108例PT-IgG检测阳性率为55.56%.101例核酸阳性病例的病程中位数为11 d;60例PT-IgG阳性病例的病程M为21 d.<1岁病例的Real-time PCR检测阳性率为84.28%,与其他年龄段阳性率比较差异有统计学意义.30份标本百日咳鲍特菌基因核苷酸同源性为99.6%～100.0%,与GenBank中国际标准株TohamaI、中国疫苗株同源性为99%.结论 2013年天津地区流行的百日咳鲍特菌与国际标准株及中国疫苗株亲缘关系较近,<1岁病例Real-time PCR检测阳性率高于其他年龄段.     

Development of a Rift Valley fever virus viremia challenge model in sheep and goats

Rift Valley fever virus (RVFV), a member of the family Bunyaviridae, causes severe to fatal disease in newborn ruminants, as well as abortions in pregnant animals; both preventable by vaccination. Availability of a challenge model is a pre-requisite for vaccine efficacy trials. Several modes of inoculation with RVFV ZH501 were tested on goats and sheep. Differences in development of infectious viremia were observed between animals inoculated with RVFV produced in mosquito C6/36 cells compared to Vero E6 cell-produced inoculum. Only C6/36-RVFV inoculation led to development of viremia in all inoculated sheep and goats. The C6/36 cell-produced RVFV appeared to be more infectious with earlier onset of viremia, especially in sheep, and may also more closely represent a field situation. Goats were somewhat more resistant to the disease development with lower and shorter infectious virus viremia, and with only some animals developing transient increase in rectal temperature in contrast to sheep. In conclusion, a challenge protocol suitable for goat and sheep vaccine efficacy studies was developed using subcutaneous inoculation of 10⁷ PFU per animal with RVFV ZH501 produced in C6/36 cells.     

Differential developmental trajectories of magnetic susceptibility in human brain gray and white matter over the lifespan

As indicated by several recent studies, magnetic susceptibility of the brain is influenced mainly by myelin in the white matter and by iron deposits in the deep nuclei. Myelination and iron deposition in the brain evolve both spatially and temporally. This evolution reflects an important characteristic of normal brain development and ageing. In this study, we assessed the changes of regional susceptibility in the human brain in vivo by examining the developmental and ageing process from 1 to 83 years of age. The evolution of magnetic susceptibility over this lifespan was found to display differential trajectories between the gray and the white matter. In both cortical and subcortical white matter, an initial decrease followed by a subsequent increase in magnetic susceptibility was observed, which could be fitted by a Poisson curve. In the gray matter, including the cortical gray matter and the iron‐rich deep nuclei, magnetic susceptibility displayed a monotonic increase that can be described by an exponential growth. The rate of change varied according to functional and anatomical regions of the brain. For the brain nuclei, the age‐related changes of susceptibility were in good agreement with the findings from R2^* measurement. Our results suggest that magnetic susceptibility may provide valuable information regarding the spatial and temporal patterns of brain myelination and iron deposition during brain maturation and ageing. Hum Brain Mapp 35:2698–2713, 2014. © 2013 Wiley Periodicals, Inc .     

Motor function and respiratory capacity in patients with late‐onset pompe disease

Introduction: The relationship between skeletal muscle strength and respiratory dysfunction in Pompe disease has not been examined by quantitative methods. We investigated correlations among lower extremity proximal muscle strength, respiratory function, and motor performance. Methods: Concentric strength of the knee extensor and flexor muscles was measured with a dynamometer, and pulmonary function was evaluated using spirometry in 7 adult patients. The 6‐minute walk test and the 4‐step stair‐climb test were used for assessing aerobic endurance and anaerobic power, respectively. Results: Anaerobic motor performance correlated with strength of both thigh muscles. Respiratory function did not correlate with either muscle strength or motor function performance. Conclusions: Respiratory and lower extremity proximal muscles could be affected differentially by the disease in individual patients. Motor performance is influenced by thigh muscle strength and is less dependent of respiratory capacity in our cohort of ambulatory patients. Muscle Nerve 49:603–606, 2014     

在β地中海贫血着床前遗传学诊断中应用多重置换扩增进行预处理的临床分析

目的分析应用多重置换扩增技术(multiple displacement amplification,MDA)进行全基因组扩增的预处理是否影响β地中海贫血着床前遗传学诊断(preimplantation genetic diagnosis,PGD)的准确效能。方法回顾性地分析2009年1月至2013年6月,因双方均为β地中海贫血携带者而行PGD治疗的周期资料,其中34个周期采用多重巢式聚合酶链反应(PCR)结合反向斑点杂交技术对单细胞进行诊断,另有38个周期行MDA进行全基因组扩增的预处理后,再结合反向斑点杂交技术进行诊断。结果两组患者在年龄、获卵数等实验室指标上无统计学差异。MDA组未检出(扩增失败)率为9.79%,低于行巢式PCR组的15.24%,而杂合子率46.33%则略高,但两种方法在诊断结果上并无统计学差异。结论应用MDA技术进行全基因组的预扩增可有效增加检测模板,实现多位点及多种疾病的诊断,而且不影响β地中海贫血地贫基因的诊断效能。     

NHS arm's length bodies and regulatory networks in England: quantitative analysis

Health regulation is an area of English public policy that involves a plethora of different bodies operating independently of one another with the purported aim of monitoring standards, ensuring minimum standards are met and providing assurance for the public and government. The purpose of this research is to ascertain as well as possible whether regulation generally is having a positive effect on service provision as well as what the relationship between the data collected by these bodies is. The rationale for testing whether the effect of regulation is positive is obvious; it costs money, time and effort, so does it work? The rationale for testing the relationships is that providers who score well on quality measures should also be experiencing fewer deaths compared with what would be expected. They should also be the providers who are performing well in terms of finances and governance. If there is no relationship between these data sets, then surely something is going wrong; that is, does the various regulatory monitoring and scrutiny actually measure what it sets out to measure? Copyright © 2013 John Wiley & Sons, Ltd.     

5-氮杂胞苷诱导大鼠心脏成纤维细胞向心肌样细胞分化进程相关基因的表达差异

目的 探讨5-氮杂胞苷（5-aza）诱导心脏成纤维细胞（CFs）向心肌样细胞分化5 d、7 d、14 d、21 d和28 d相关基因的表达差异。方法 新生1～3 d SD大鼠12只,胰蛋白酶/胶原酶联合消化法分离培养新生大鼠CFs,采用CFs分子标记盘状结构域受体2（DDR2）并行细胞免疫荧光染色。含15 μmol/L 5-aza心肌细胞诱导液培养第3代CFs, Real-time PCR检测心肌化诱导不同时间点相关基因vimentin、DDR2、Nanog、sox2、c-kit、sca-1、Tbx5、CD73、CD34、cMyc、klf4、Gata4、Oct4、Mef2c和心肌肌钙蛋白T（cTnT）的表达差异,细胞免疫荧光检测心肌化诱导后5 d、7 d、14 d、21 d和28 d肌钙蛋白T（cTnT）的表达。透射电子显微镜观察心肌化诱导28 d CFs的超微结构变化。结果 5-aza诱导CFs后3 d 细胞生长速率减慢,部分细胞由三角形、梭形向圆形和杆状转变。诱导后7 d、14 d、21 d 和28 d,与诱导前相比,DDR2表达稳定,vimentin在14 d表达下降,Nanog、c-kit、sox2和Tbx5伴随着心肌化进程呈现表达下降趋势,而CD73、Mef2c、CD34、Gata4、Oct4和cTnT表达逐步增加,sca-1在7 d 表达上升又下降,cMyc和klf4在14 d表达上升又下降。诱导后 5 d少量细胞表达cTnT,14～21 d cTnT阳性细胞数明显增高,28 d表达cTnT较多且趋于稳定。透射电子显微镜显示,CFs心肌化诱导28 d,细胞与细胞间出现端-端连接,细胞质内出现大量的肌原纤维,排列较为规律,但肌节不明显,H带、Z线和M线显示不清。结论 CFs经5-aza诱导,可向心肌样细胞分化,但不能形成成熟的心肌细胞。     

微小RNA-423-5p靶向乙醛脱氢酶2减轻脂多糖诱导的人脐静脉血管内皮细胞损伤

目的 探讨微小RNA-423-5p(miR-423-5p)对脂多糖（LPS）诱导血管内皮细胞损伤的保护及作用机制。方法 用1 mg/L LPS诱导人脐静脉血管内皮细胞（HUVECs）24 h,Real-time PCR和Western blotting检测细胞中miR-423-5p和乙醛脱氢酶2（ALDH2）的表达。通过转染anti-miR-423-5p和pcDNA-ALDH2下调miR-423-5p和上调ALDH2表达,流式细胞术检测细胞凋亡率,Western blotting检测凋亡相关蛋白Bcl-2和Bax的表达,并用ELISA试剂盒检测LPS诱导后细胞上清液中白细胞介素6（IL-6）和肿瘤坏死因子α（TNF-α）的含量;双荧光素酶报告系统验证miR-423-5p与ALDH2的调控关系。结果 与对照相比,LPS可诱导HUVECs凋亡和损伤,使HUVECs中miR-423-5p、Bax表达量及IL-6和TNF-α分泌量均显著升高（P<0.05）,ALDH2的mRNA和蛋白表达量及Bcl-2量显著降低（P<0.05）;下调mi-423-5p表达和过表达ALDH2均可减轻LPS诱导的HUVECs损伤并抑制细胞凋亡;miR-423-5p靶向负调控ALDH2的表达;抑制ALDH2表达逆转了下调miR-423-5p表达对LPS诱导的HUVECs损伤的作用。结论 下调miR-423-5p表达可靶向ALDH2减轻LPS对HUVECs的损伤并抑制细胞凋亡。     

醉茄素A通过诱导白色脂肪组织褐变抵御高脂饮食诱导的肥胖

目的 探索醉茄素A对高脂饮食诱导肥胖的抵抗作用及其作用机制。方法 8周龄C57BL/6 J小鼠分为醉茄素A组和对照组,每组10只,高脂饮食（HFD）饲养,分别给予醉茄素A或对照溶剂DMSO腹腔注射,每天测量小鼠体重及摄食量。1周后取材,称量小鼠腹股沟白色脂肪（iWAT）、附睾白色脂肪（eWAT）、腹膜后白色脂肪（rWAT）和肩胛间棕色脂肪（BAT）的重量;取iWAT检测褐变相关因子表达水平;HE染色观察iWAT细胞形态变化。结果 醉茄素A组小鼠的体重和各部位脂肪组织重量均显著低于对照组,摄食量无明显变化;Real-time PCR显示,醉茄素A组小鼠iWAT中褐变相关基因表达水平显著提高;Western blotting结果表明,醉茄素A组小鼠iWAT中解耦联蛋白1（UCP1）和过氧化物酶体增殖体激活受体γ共激活因子1α（PGC1α）表达明显增加;HE染色与免疫荧光结果显示,醉茄素A组小鼠腹股沟白色脂肪细胞呈现典型褐变特征,如脂肪细胞多腔室化。结论 醉茄素A可能通过诱导小鼠白色脂肪组织褐变抵御高脂饮食诱导的肥胖。