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免脱盐柱除盐的蛋白组学前处理方法

实验室科学 2024年第5期27卷 33-35,40页

作者：季学猛史爱莹吴景张燕王硕南开大学医学院天津300071

现有的蛋白组样品前处理中脱盐柱除盐的方法存在步骤繁多、易损失微量样品等缺点,这里,一种超滤管辅助酶解及加热潮解除盐的蛋白组学前处理方法被提出。具体而言,是利用超滤管辅助,通过离心在酶解前将缓冲液置换成碳酸氢铵溶液,酶解后... 详细信息

现有的蛋白组样品前处理中脱盐柱除盐的方法存在步骤繁多、易损失微量样品等缺点,这里,一种超滤管辅助酶解及加热潮解除盐的蛋白组学前处理方法被提出。具体而言,是利用超滤管辅助,通过离心在酶解前将缓冲液置换成碳酸氢铵溶液,酶解后多肽溶液中的碳酸氢铵被加热除去,最终得到脱盐的纯净多肽。该方法操作简便,不损失样品,而且无需洗脱步骤,可以方便地与LC-MS直接衔接,进行质谱分析。

关键词：蛋白组学样品前处理脱盐柱加热碳酸氢铵

Diagnostic value of serum human epididymis protein 4 in esophageal squamous cell carcinoma

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World Journal of Gastrointestinal Oncology 2020年第10期12卷 1167-1176页

作者： shi-Yuan Liu Muhammad Ahsan Bilal Jian-hong Zhu Shao-Min Li Department of Thoracic Surgery Second Affiliated Hospital of Xi'an Jiaotong UniversityXi'an 710004Shaanxi ProvinceChina Department of Dermatology and Venereology Second Affiliated Hospital of Xi'an Jiaotong UniversityXi'an 710004Shaanxi ProvinceChina Department of Clinical Laboratory Second Affiliated Hospital of Xi'an Jiaotong UniversityXi'an 710004Shaanxi ProvinceChina

BACKGROUND Numerous studies have demonstrated that human epididymis protein 4(HE4)is overexpressed in various malignant tissues including ovarian,endometrial,lung,breast,pancreatic,and gastric ***,no study has examined the diagnostic impact of HE4 in patient with esophageal squamous cell carcinoma(ESCC)until *** To analyze the value of four serum tumor markers for the diagnosis of ESCC,and examine the associations of serum levels of HE4 with ESCC patients’clinicopathological *** The case group consisted of 80 ESCC patients,which were compared to a control group of 56 patients with benign esophageal *** levels of HE4,carcinoma embryonic antigen(CEA),alpha fetal protein,and carbohydrate antigen 19-9(CA19-9)were detected by *** associations of serum HE4 levels with ESCC patients’clinicopathological characteristics such as gender,tumor location,and pathological stage were also examined after *** The result of ELISA showed that serum HE4 level was significantly higher in the patients with ESCC than in the controls,and the staining intensity was inversely correlated with the pathological T and N *** HE4 levels had a sensitivity of 66.2%and specificity of 78.6%when the cutoff value was set at 3.9 ng/***,the combined HE4 and CA19-9 increased the sensitivity to 83.33%,and interestingly,the combination of HE4 with CEA led to the most powerful sensitivity of 87.5%.Furthermore,A positive correlation was observed between HE4 serum levels and pathological T and N stages(P=0.0002 and 0.0017,respectively),but there was no correlation between HE4 serum levels and ESCC patient gender(P=0.4395)or tumor location(P=0.6777).CONCLUSION The results of this study suggest that detection of serum HE4 levels may be useful in auxiliary diagnosis and evaluation of the progression of ESCC.

关键词： Human epididymis protein 4 Esophageal squamous cell carcinoma Diagnostic value Histological stage Clinicopathological characteristics Clinical progression

Novel mutation in a Chinese patient with progressive familial intrahepatic cholestasis type 3

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World Journal of Gastroenterology 2015年第2期21卷 699-703页

作者： Hao-Zhe Sun hong shi Shun-Cai Zhang Xi-Zhong Shen Department of Gastroenterology and Hepatology Zhongshan HospitalFudan University Shanghai Medical College Fudan University

Genotyping is conclusive for the diagnosis of progressive familial intrahepatic cholestasis type 3(PFIC3). Here we report a Chinese patient of PFIC3 with compound mutations in the ABCB4 gene. Liver biopsy was performed on a 17-year-old male patient with intrahepatic cholestasis of unknown etiology. Liver histology findings are indicative of intrahepatic cholestasis with extensive fibrosis. Genotyping revealed c.175C>T(p.L59L) mutation in exon 4, c.504C>T(p.N168N) mutation in exon 6, c.711A>T(p.I237I) mutation in exon 8, c.874A>T(p.K292X) in exon 9 and a novel mutation, c.1804G>T(p.G602W) in exon 15. Based on these findings, the patient was diagnosed with PFIC3. The novel mutation p.G602 W in exon 15 was predicted as probably damaging by Poly Phen-2 with a score of 0.986(sensitivity: 0.54; specificity: 0.94) and was predicted to affect protein function with a SIFT score of 0.01.

关键词： Liver disease Cholestasis Progressive familial int

Base cation concentrations in forest litter and topsoil have different responses to climate and tree species along elevational gradients

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Journal of Mountain Science 2019年第1期16卷 30-42页

作者： XUE Feng ZHAO Ming-fei WANG Yu-hang KANG Mu-yi XING Kai-xiong WANG Guo-yi shi jing-jing CHEN Chen JIANG Yuan Beijing Key Laboratory of Traditional Chinese Medicine Protection and Utilization Faculty of Geographical Science Beijing Normal University College of Resources Science and Technology Faculty of Geographical Science Beijing Normal University Institute of Geographic Sciences and Natural Resources Research Chinese Academy of Sciences Faculty of Natural Resources Management Lakehead University

The forest litter is an essential reservoir of nutrients in forests, supplying a large part of absorbable base cations(BC) to topsoil, and facilitating plant growth within litter-soil system. To characterize elevational patterns of base cation concentrations in the forest litter and topsoil, and explore the effects of climate and tree species, we measured microclimate and collected the forest litter and topsoil(0-10 cm) samples across an elevational range of more than 2000 m(1243 ～ 3316 m a.s.l.),and analyzed the concentrations of BC in laboratory. Results showed that: 1) litter Ca concentration displayed a hump-shaped pattern along the elevational gradients, but litter K and Mg showed saddle-shaped patterns. Soil Ca concentration increased with elevation, while soil K and Mg had no significant changes. 2) Ca concentration in the forest litter under aspen(Populus davidiana) was significantly higher than that in all other species, but in topsoil, Ca concentration was higher under coniferous larch and fir(Larix chinensis and Abies fargesii). Litter K and Mg concentrations was higher under coniferous larch and fir, whereas there were nosignificant differences among tree species in the concentrations of K and Mg in topsoil. 3) Climatic factors including mean annual temperature(MAT), growing season precipitation(GSP) and non-growing season precipitation(NGSP) determined BC concentrations in the forest litter and topsoil. Soil C/N and C/P also influenced BC cycling between litter and soil. Observation along elevations within different tree species implies that above-ground tree species can redistribute below-ground cations, and this process is profoundly impacted by climate. Litter and soil Ca, K and Mg with different responses to environmental variables depend on their soluble capacity and mobile ability.

关键词： Base cations Elevations Larix chinensis Litter-soil system Carbon nitrogen ratio

3643例合肥地区住院儿童13种呼吸道病原体的流行病学分析

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标记免疫分析与临床 2024年第2期31卷 232-236页

作者：吴静石玉如刘婷秦令寒戚应杰中国科学技术大学附属第一医院(安徽省立医院)感染病院检验科安徽合肥230000

目的了解分析合肥地区住院儿童13种呼吸道病原体感染情况。方法选取2020年9月至2023年9月安徽省立医院感染病院呼吸道病毒检测标本3643例,采用多重反转录聚合酶链反应(RT-PCR)技术扩增了解13种呼吸道感染病毒:甲型流感病毒(InfA)、甲型... 详细信息

目的了解分析合肥地区住院儿童13种呼吸道病原体感染情况。方法选取2020年9月至2023年9月安徽省立医院感染病院呼吸道病毒检测标本3643例,采用多重反转录聚合酶链反应(RT-PCR)技术扩增了解13种呼吸道感染病毒:甲型流感病毒(InfA)、甲型流感病毒H1N1病毒(H1N1)、甲型流感病毒H3N2(H3N2)、副流感病毒(HPIV)、偏肺病毒(HMPV)、肺炎支原体(MP)、呼吸道合胞病毒(HRSV)、腺病毒(HADV)、鼻病毒(HRV)、博卡病毒(Boca)、衣原体(Ch)、乙型流感病毒(InfB)和冠状病毒(HCOV)的流行病学特点。结果3643例标本中阳性标本共计2222例,阳性率为60.99%,354例为混合病毒感染,多重感染阳性率为9.72%,1868例为单一病毒感染(除InfA未检出外),阳性率为51.27%。1299例男性呼吸道病毒检测阳性(60.96%),923例女性呼吸道病毒检测阳性(61.04%),男女患儿感染的阳性率差异无统计学意义,感染患者主要集中于<1岁组(61.94%)。春季以HRV检出为主(20.50%),夏季以MP和HRV检出为主(24.79%和12.39%),秋季以HRSV和MP检出为主(15.09%和14.30%),HRSV在冬季检出率最高(19.05%)。结论呼吸道病毒感染在合肥住院儿童中非常普遍,近年来MP、HRV和HRSV是非常突出的呼吸道感染的重要病原体,呼吸道病毒感染与患儿年龄和季节分布有关,这些发现可能为合肥住院儿童呼吸道易感染患者的临床诊断治疗及合理用药提供有力的证据。

关键词：呼吸道病毒儿童流行特征

非洲猪瘟病毒C84L蛋白通过激活NLRP3炎症小体上调炎症因子的表达

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中国兽医科学 2024年第1期54卷 1-11页

作者：唐静马旭升石正旺代军飞叶得河王萌马永华郑海学甘肃农业大学动物医学院甘肃兰州730070 中国农业科学院兰州兽医研究所兰州大学动物医学与生物安全学院动物疫病防控全国重点实验室甘肃兰州730000

探索非洲猪瘟病毒(African swine fever virus,ASFV)调控细胞因子产生和细胞死亡的机制。利用NLRP3炎症小体表达系统,筛选发现ASFV-C84L蛋白诱导NLRP3炎症小体介导的IL-1β的分泌上调。首先,利用生物信息学分析C84L的结构信息,通过实时... 详细信息

探索非洲猪瘟病毒(African swine fever virus,ASFV)调控细胞因子产生和细胞死亡的机制。利用NLRP3炎症小体表达系统,筛选发现ASFV-C84L蛋白诱导NLRP3炎症小体介导的IL-1β的分泌上调。首先,利用生物信息学分析C84L的结构信息,通过实时荧光定量PCR(qPCR)和间接免疫荧光试验(IFA)确定C84L蛋白在ASFV感染后的表达时序,细胞内定位;同时,用荧光素酶活性检测和蛋白免疫印迹(Western-blot)确定C84L蛋白参与调控p65的磷酸化以及IL-1β的成熟和细胞焦亡;然后,用碘化丙啶(PI)染色观察C84L蛋白诱导细胞的死亡情况;最后,使用qPCR和酶联免疫吸附试验检测C84L蛋白对IL-1β、IL-6、TNF-α等细胞因子转录和分泌的影响。结果显示:C84L为亲水性蛋白,与沙门菌效应蛋白Sif A具有同源性;ASFV感染原代猪肺泡巨噬细胞4 h后,C84L m RNA表达水平逐渐上调,后续持续表达,第8小时达到顶峰;而且IFA结果显示,C84L蛋白在细胞质和细胞核中均有定位。荧光素酶和Western-blot结果显示,C84L促进p65的磷酸化并激活NF-κB启动子的活化。PI染色结果显示,C84L真核质粒转染细胞后诱导细胞死亡,Western-blot结果也显示C84L诱导Caspase-1成熟以及膜孔蛋白N-GSDMD (GSDMD剪切后的N端片段)剪切。将C84L真核质粒转染细胞后,IL-1β、IL-6、TNF-α等细胞因子转录水平呈剂量依赖性升高;与转录水平相似,C84L表达诱导IL-1β、IL-6、TNF-α分泌水平上调。结论:ASFV C84L蛋白通过激活NF-κB以及NLRP3炎症小体诱导IL-1β、IL-6、TNF-α等促炎细胞因子表达水平上调以及细胞焦亡的发生。

关键词：非洲猪瘟病毒细胞因子 C84L NF-κB NLRP3

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便携式联用技术快速鉴别铜冶炼烟尘

中国口岸科学技术 2024年第5期6卷 59-64页

作者：廖敏萍王石陈永欣杨晶丽吴国境张博阮贵武防城海关综合技术服务中心防城港538001 防城海关防城港538000 中国检验认证集团广西有限公司防城港分公司防城港538000 广西金川有色金属有限公司防城港538000

铜冶炼烟尘与铜精矿由相似元素组成,通过元素种类与含量一般难以对两者进行准确鉴别。我国铜精矿进口量的日益增加及物料性质趋于复杂,为货物的通关效率及现场查验带来挑战,因此解决现场进口铜矿的固废快速筛查显得尤为重要。本文以进... 详细信息

铜冶炼烟尘与铜精矿由相似元素组成,通过元素种类与含量一般难以对两者进行准确鉴别。我国铜精矿进口量的日益增加及物料性质趋于复杂,为货物的通关效率及现场查验带来挑战,因此解决现场进口铜矿的固废快速筛查显得尤为重要。本文以进口典型铜精矿及主要的铜火法冶炼烟尘作为研究样品,联合使用手持式X射线荧光光谱仪和便携式X射线衍射仪对样品中的物质组成和结构特征进行解析,探索铜精矿与铜火法冶炼烟尘的主要区别点,建立了铜冶炼烟尘现场快速属性识别方法。

关键词：铜冶炼烟尘便携式 X射线荧光光谱 X射线衍射

通督益脑针法联合化痰通络方治疗缺血性脑卒中假性球麻痹吞咽障碍患者的疗效及对其血清脑源性神经营养因子、类胰岛素一号生长因子含量的影响

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世界中西医结合杂志 2024年第6期19卷 1196-1200,1206页

作者：周艳朋刘晶晶焦雪蕾张福磊刘宏伟张小健北京中医医院顺义医院脑病科北京101300

目的观察通督益脑针法联合化痰通络方治疗缺血性脑卒中假性球麻痹吞咽障碍患者的疗效及对其血清脑源性神经营养因子(Brain-derived neurotrophic factor,BDNF)及类胰岛素一号生长因子(Insulin-like growth factor I,IGF-1)含量的影响。... 详细信息

目的观察通督益脑针法联合化痰通络方治疗缺血性脑卒中假性球麻痹吞咽障碍患者的疗效及对其血清脑源性神经营养因子(Brain-derived neurotrophic factor,BDNF)及类胰岛素一号生长因子(Insulin-like growth factor I,IGF-1)含量的影响。方法选取2020年12月—2021年12月期间北京中医医院顺义医院神经内科收治的110例缺血性脑卒中假性球麻痹吞咽障碍患者为研究对象,按随机数字表法分为对照组和观察组,每组各55例。对照组采用常规治疗及康复锻炼,观察组在对照组基础上予以通督益脑针法联合化痰通络方治疗,均治疗4周。观察比较两组患者临床疗效,治疗前后中医症状积分、吞咽功能[吞咽功能评定量表(Standardizedswallowing assessment,SSA)及洼田吞咽能力评定量表]、吞咽困难特异性生活质量量表(Swallowquality-of-life questionnaire,SWAL-QOL)、BDNF及IGF-1含量。结果治疗后观察组总有效率92.73%(51/55)明显高于对照组80.00%(44/55),差异有统计学意义(P<0.05)。治疗后两组患者SSA、洼田饮水评分均较治疗前降低,SWAL-QOL评分均较治疗前升高,差异有统计学意义(P<0.05);且观察组SSA、洼田饮水评分均明显低于对照组,SWAL-QOL评分明显高于对照组,差异有统计学意义(P<0.05)。治疗后两组患者中医证候积分均较治疗前下降,差异有统计学意义(P<0.05);且观察组中医证候积分明显低于对照组,差异有统计学意义(P<0.05)。治疗后两组患者血清BDNF及IGF-1含量均较治疗前明显升高,差异有统计学意义(P<0.05);且观察组血清BDNF及IGF-1含量均明显高于对照组,差异有统计学意义(P<0.05)。结论通督益脑针法联合化痰通络方能够显著改善缺血性脑卒中假性球麻痹吞咽障碍患者神经功能,上调神经因子水平,减轻吞咽困难症状,改善其生活水平,疗效显著,值得临床推广。

关键词：通督益脑针法化痰通络方缺血性脑卒中假性球麻痹吞咽障碍

Protective Effect of Norcantharidin on Collagen-Induced Arthritis Rats

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Chinese Journal of Integrative Medicine 2018年第4期24卷 278-283页

作者： SHEN hong-bo HUO Ze-jun BAI Yun-jing HE Xiao-juan LI Chang-hong ZHAO Yu-kun GUO Qing-qing Department of Traditional Chinese Medicine Peking University Third Hospital Division of Rheumatology PLA Army General Hospital Institute of Basic Research in Clinical Medicine Academy of Chinese Medical Sciences Division of Rheumatology Peking University Third Hospital

Objective： To observe the effect of norcantharidin（NCTD） on collagen-induced arthritis（CIA） rats. Methods： Sixty Sprague-Dawley（SD） rats were randomly divided into 6 groups（n=10）： normal group, CIA model group（model group）, NCTD low-dose group [1.35 mg/（kg·d）], NCTD middle-dose group [2.7 mg/（kg·d）], NCTD high-dose group [5.4 mg/（kg·d）] and methotrexate（MTX） group [1.8 mg/（kg/w）]. Anesthetized rats were sacrificed by luxation of cervical vertebra after 4 weeks of administration. The arthritis scores were evaluated twice a week. The pathological changes in the ankle joints of rats were observed by hematoxylin-eosin（H＆E） staining. The serum levels of interleukin（IL） 1β, IL-6, tumor necrosis factor（TNF）-α, vascular endothelial growth factor（VEGF）, IL-17 and transform growth factor（TGF） β were detected by enzyme linked immunosorbent assay（ELISA）. The mRNA expression of retinoid-related orphan nuclear receptor γ t（ROR γ t） and forkhead box P3（Foxp3） in peripheral blood lymphocytes were confirmed by real-time polymerase chain reaction. Results： MTX and high-dose NCTD not only decreased the arthritis scores but also alleviated the pathological changes in CIA rats＇ ankle joints compared with the model group（P〈0.05 or P〈0.01）. All doses of NCTD significantly inhibited the serum levels of IL-6, IL-17 and TNF-α in CIA rats（P〈0.05）. Only middle-and high-dose of NCTD prominently decreased serum IL-1β and TGF-β levels of CIA rats（P〈0.05）. However, NCTD has no effect on vascular endothelial growth factor（VEGF） level in CIA rats. The Foxp3 mRNA expression in all NCTD groups were increased significantly than in the model group（P〈0.05）. The mRNA expression of RORγt in NCTD high-dose group was decreased apparently in comparison with the model group（P〈0.05）. Conclusion： NCTD showed therapeutic effect on CIA rats by inhibition of cytokines and regulation of Th17/Treg cells.

关键词： norcantharidin collagen-induced arthritis cytokine Th17/Treg forkhead box P3 retinoid related orphan nuclear receptor γt