支气管哮喘患者外周血Th17细胞及相关趋化因子表达水平检测

目的探讨支气管哮喘患者外周血Th17细胞及相关趋化因子表达水平与肺功能的关系。方法选取60例支气管哮喘患者分为急性发作组和缓解组,每组各30例,另选取健康体检者30例为对照组。采用流式细胞术检测三组人群外周血中Th17细胞(CD4+IL-17+)上CCR4、CCR6表达水平,用ELISA法检测患者血清中总IgE、IL-17、IL-23、CCL20的蛋白表达水平。结果 1急性发作组哮喘患者外周血Th17细胞CCR4、CCR6表达水平明显高于缓解组哮喘患者(P<0.05),缓解组哮喘患者外周血Th17细胞CCR4、CCR6表达水平高于健康对照组(P<0.05);2急性发作组哮喘患者外周血血清中总IgE、IL-17、IL-23、CCL20的蛋白表达水平明显高于缓解组哮喘患者(P<0.05),缓解组哮喘患者外周血血清中总IgE、IL-17、IL-23、CCL20的蛋白表达水平高于健康对照组(P<0.05);3CCR4、CCR6表达水平与患者的总IgE、IL-17、IL-23、CCL20的蛋白表达水平正相关(P<0.05)。结论支气管哮喘患者外周血中Thl7细胞相关趋化因子CCR4、CCR6表达增高并与Th17细胞造成的炎症损伤有关。     

NLRP7在早期葡萄胎诊断中的应用

目的：探讨NLRP7、P57和Ki－67免疫组织化学染色在早期葡萄胎的表达情况及应用价值。方法根据葡萄胎病理学诊断标准结合P57的表达,将确诊的葡萄胎分为部分性（PHM）和完全性葡萄胎（CHM）,利用免疫组织化学染色检测妊娠早期的部分性葡萄胎（n＝22）和完全性葡萄胎（n＝24）石蜡组织切片中NLRP7和Ki－67蛋白的表达。结果 NLRP7蛋白在部分性葡萄胎和完全性葡萄胎的表达率分别为54．55％（12／22）和50％（12／24）,差异无统计学意义（ P＞0．05）,但CHM组NLRP7蛋白的表达强度组内比较有统计学差异,且强阳性表达组和部分性葡萄胎组相比,有显著统计学差异（P＜0．01）。 Ki－67在部分性和完全性葡萄胎组表达率分别为59．09％（13／22）和79．17％（19／24）,有统计学差异（P＜0．05）。结论 NL-RP7蛋白单项检测不可直接用于鉴别部分性和完全性葡萄胎,需进一步检测基因突变,但联合P57和Ki－67蛋白可为复发性葡萄胎的预防及监测提供指导意义。     

阿利吉仑对大鼠肾纤维化TGF-β1/Smad信号通路的影响

目的探讨阿利吉仑对单侧输尿管梗阻大鼠肾间质纤维化的作用及机制。方法 24只雌性SD大鼠随机分为3组,Sham组、UUO组及aliskiren组,除Sham组外,UUO组及aliskiren组均行左侧输尿管结扎术。aliskiren组术前1 d开始灌胃给药,阿利吉伦50 mg/(kg·d),每日1次,连续2周。术后第14天处死大鼠,留取梗阻侧肾组织行HE染色和Masson染色,光镜下观察各组大鼠肾组织病理变化,免疫组化和Western印迹检测肾组织TGF-β1、Smad2和Smad7蛋白表达水平。结果阿利吉仑可明显减轻大鼠肾间质炎性细胞浸润、纤维组织增生及肾小管扩张和萎缩。免疫组化和Western印迹结果均显示:与Sham组相比,UUO组TGF-β1、Smad2表达明显上升,而Smad7表达明显下降(均P<0.05);与UUO组相比,aliskiren组TGF-β1、Smad2表达明显下降,Smad7表达明显上升(均P<0.05)。结论 TGF-β1/Smad信号通路与肾间质纤维化有关,阿利吉仑可抑制肾间质纤维化,其机制可能与下调TGF-β1、Smad2的表达和上调Smad7的表达有关。     

纳米雄黄对人宫颈癌细胞增殖和凋亡的影响

目的：探讨纳米雄黄对不同宫颈癌细胞株Hela(HPV18阳性,腺癌),Caski(HPV16阳性,鳞癌), C33A(HPV阴性,腺癌)增殖、凋亡的影响。方法：体外培养3种不同宫颈癌细胞,采用不同质量浓度(5,10, 20,40 mg/L)纳米雄黄作用于不同宫颈癌细胞24,48,72,96 h,相差显微镜下观察不同质量浓度组与对照 组细胞形态学变化;MTT法测定药物对细胞生长的抑制作用;流式细胞仪检测细胞凋亡及细胞周期。结果： 纳米雄黄混悬液(5,10,20,40 mg/L)作用于Caski,Hela,C33A细胞24,48,72,96 h,均可抑制细胞增殖, 且呈时间剂量依赖性,抑制率波动于9.02%~49.06%,以20 mg/L质量浓度组为例,抑制率Caski(39.15%)>Hela (36.17%)>C33A(30.56%)。随着质量浓度的增加,早期凋亡、中晚期凋亡细胞均增加,凋亡率(早期+中晚期凋亡率)波 动于19.29%~99.54%,以20 mg/L质量浓度组为例,凋亡率Caski(60.43±2.88)%>Hela (41.95±3.01)%>C33A (43.49±2.19)%。 细胞周期结果显示高质量浓度组(20,40 mg/L)对Hela,Caski细胞有G2/M期阻滞。结论：纳米雄黄混悬液可抑制不同 宫颈癌细胞株的增殖,促进细胞凋亡,其对Caski细胞的作用最强,对Hela细胞次之,对C33A细胞相对最弱。对HPV 阳性的细胞可产生G2/M期阻滞。     

近红外发光葡聚糖纳米胶活体成像小鼠淋巴结

 目的  制备和研究荧光染料标记的葡聚糖纳米胶,用于活体荧光成像小鼠腋窝淋巴结。方法  使用脱氧胆酸和荧光染料Cy7化学标记葡聚糖,通过透析法制备纳米胶Dextran-Cy7;使用透射电子显微镜、动态光散射仪分别表征纳米胶的形貌及大小;分光光度法标定Cy7的含量;用C57小鼠和DC2.4细胞分别评价Dextran-Cy7的体内、体外毒性;对10只正常昆明小白鼠经前掌注射Dextran-Cy7生理盐溶液,使用活体荧光扫描成像系统观察小鼠腋窝淋巴结的成像效果;淋巴结免疫荧光组化法观察Dextran-Cy7在淋巴结内的分布。结果  Dextran-Cy7纳米胶是直径30~50 nm的球形纳米颗粒,其在体内和体外都具有较低的细胞毒性,可作为近红外发光探针活体荧光成像小鼠腋窝淋巴结;纳米胶主要分布在淋巴结的巨噬细胞细胞内。结论  Dextran-Cy7纳米胶细胞毒性低、生物相容性好,可作为纳米荧光探针成像活体小鼠淋巴结。     

中国人感染H7N9禽流感危险区域预警识别的研究

目的 建立人感染H7N9禽流感发病危险预警模型, 识别禽流感暴发高危险区域并提出预警。方法 收集2013年2月至2014年6月中国地市级人感染H7N9禽流感病例数据及同期地理、气象数据, 通过空间自回归(SAR)模型和广义相加模型(GAM)拟合并量化地理和气象因素对发病的影响, 综合两模型的预测结果建立发病危险预警地图。结果 2014年2月中国人感染H7N9禽流感的实际发病地区全部位于研究构建的发病危险预测区域内;模型预测了疾病的空间移动趋势, 对2014年4、5月北方地区的新发疫情有准确的预警。结论 建立的模型短期预测准确度和精确度较好, 可应用于疫情监测和预警领域, 有助于早期区域预防疫情的流行及暴发。     

Early impact of PCV7/PCV13 sequential introduction to the national pediatric immunization plan,on adult invasive pneumococcal disease: A nationwide surveillance study

BackgroundPCV7 was introduced as a universal childhood vaccination in Israel on July 2009 and was gradually replaced by PCV13 from November 2010. We report data on adult invasive pneumococcal disease (IPD), two years post PCV13 implementation.MethodsAn ongoing nationwide active surveillance (all 27 laboratories performing blood/CSF cultures nationwide), initiated in 2009, providing all blood/CSF Streptococcus pneumoniae isolated from persons ≥18 years. Capture-recapture method assured reporting of >95% cases. All isolates were serotyped in one central laboratory. Medical history and outcomes were recorded in ∼90%.ResultsOf 1809 IPD episodes, S. pneumoniae was isolated from the blood in 95% and most cases had pneumonia. Predisposing comorbidities were present in >70%. During the four study years, overall IPD incidence decreased from 9.2 to 7.2/100,000, incidence of pneumonia and particularly severe pneumonia cases decreased significantly from 6.6 to 4.7/100,000, (p = 0.029). Vaccine type (VT7/VT13) serotypes decreased by 70%/57% within 4 years. This was accompanied by a 52% increase in non-VT13 strains. These changes were most apparent in winter. PCV impact was most pronounced in younger adults (39% decrease in overall IPD with only a non-significant increase in non-VT13 cases) while in those >65 years a non-significant decrease in overall IPD was observed with a 64% increase in non-VT13 cases. Non-VT13 serotypes that increased significantly were 12F, 15A 10A and 6 C. A continuous reduction in isolates with penicillin MIC > 0.06 μg/ml was observed (26% to 11%, p < 0.001).ConclusionsFour years after PCV7 and 2.5 years after PCV13 universal implementation in children, incidence of adult IPD caused by VT7 and VT13 decreased in all ages, mainly in younger adults. Despite increase in non-VT13 IPD, overall IPD decreased. Additional follow-up is needed to determine the long-term impact of PCV13.     

Measuring the effects of pneumococcal conjugate vaccine (PCV7) on Streptococcus pneumoniae carriage and antibiotic resistance: The Palestinian-Israeli Collaborative Research (PICR)

BackgroundThe Palestinian-Israeli Collaborative Research (PICR) cross-conflict setting provided a unique opportunity to study overall and indirect effects of pneumococcal conjugate vaccine (PCV7), in two closely related Palestinian populations governed by two distinct health authorities with distinct vaccination policies. Here, PCV7 effects on pneumococcal carriage, serotype distribution and antibiotic resistance are reported.MethodsAnnual cross-sectional surveys of pneumococcal carriage were performed during 2009–2011 among Palestinian children (≤5 years) (a) under Palestinian-Authority (PA) health policy (Ramallah, Nablus and Bethlehem), where PCV7 was unlicensed (b) under Israeli health policy (East-Jerusalem (EJ)) where PCV7 was rapidly implemented from July 2009. Clinical data were collected, pneumococci identified and characterized for antibiotic susceptibilities and serotype. Analyses included multivariate logistic models with an interaction term for PCV7-effect.ResultsAltogether, 2755 children from PA (n = 1772) and EJ (n = 983) were enrolled, of which ∼30% were pneumococcal carriers. While overall carriage was not affected by vaccination policy, carriage of vaccine-type (VT7) strains decreased from 52% to 22% (p < 0.001) in EJ, where PCV was implemented, but not in PA. This was accompanied by an increase in non-VT13 strains from 34% to 65% (p < 0.001) in EJ, but not in PA. Furthermore, within two years post-PCV7 introduction, proportion of multi-drug resistant strains, which was initially 23% in both populations, decreased significantly in EJ, to 10%, while simultaneously it increased in PA to 33% (p < 0.001). Similar trends were observed for resistance to most antibiotic groups. The proportion of resistant isolates among non-VT13 strains did not change during the study period.ConclusionsThe unique study design distinguishes secular and seasonal effects from true vaccine effects. While PCV7 did not affect overall pneumococcal carriage rate, VT7 strains, many of which were antibiotic resistant decreased and were replaced by non-VT13 strains, which were mostly not antibiotic resistant, resulting in a net decrease in antibiotic resistance.     

基于上转发光免疫层析技术的常见食源性致病菌快速检测方法研究与评价

目的：研制针对4种常见食源性致病菌：甲型副伤寒沙门菌（ Salmonella paratyphi A）、乙型副伤寒沙门菌（Salmonella paratyphi B）、大肠埃希菌O157∶H7（Escherichia coli O157∶H7）、副溶血弧菌（Vibrio parahaemolyticus）的基于上转发光免疫层析技术（ UPT－LF）的快速定量检测试纸,并对其检测性能进行评价。方法以上转发光纳米颗粒（ UCP－NPs）作为示踪物,基于双抗体夹心检测模型,研制针对上述4种靶标菌的UPT－LF试纸;以4种靶标菌的系列浓度标准菌悬液作为标准样品,评价试纸的敏感性、特异性、线性和精密性,并进行模拟染菌食品的检测,评价模拟样品阳性检出率。结果针对4种靶标菌的UPT－LF试纸均具有良好的线性,相关系数r为0．985～0．996;检测敏感性达105～106 CFU／ml,与其他近缘菌株无交叉反应;对乳制品、饮料、小食品、水产、肉类等细菌污染的食品检出率较高。结论该研究建立的对4种常见食源性致病菌进行快速检测的UPT－LF,简便快速,具有良好的敏感性、特异性和线性定量能力,操作性能可满足食品安全检测的要求。