化学发光蛋白芯片用于检测肝癌患者血清CA19-9水平的方法研究

目的:探讨一种简单、有效、低成本、低耗时的化学发光蛋白芯片方法用于检测血清中的糖链抗原19-9(Carbohydrate Antigen19-9,CA19-9),以有助于对原发性肝癌的早期辅助诊断。方法:预先在醛基芯片上包被鼠源CA19-9单克隆抗体,建立CA19-9抗体蛋白芯片,共筛选出46份肝癌血清和32份正常健康人血清,然后用蛋白芯片方法进行检测,并以化学发光成像对检测结果进行判定。结果:24份肝癌血清的CA19-9水平高于37 U/m L,22份肝癌血清的CA19-9水平低于37 U/m L;30份正常人血清CA19-9含量低于37 U/m L,2份正常人血清的CA19-9含量为37 U/m L;灵敏度为52.17%,特异性为93.75%,ROC曲线下面积0.688[95%CI:0.566,0.811]。结论:本研究成功的建立了血清CA19-9的化学发光蛋白芯片检测方法。     

癌胚抗原蛋白芯片的研发及其在肝癌检测中的评价

目的:研究建立一种简便、快速、特异性高、低成本的检测血清中癌胚抗原(CEA)浓度的蛋白芯片,并通过检测肝细胞肝癌(HCC)对其进行评价。方法:采用双抗体夹心法,制备能够形成捕获抗体-抗原-检测抗体的"三明治"结构的蛋白芯片检测血清中CEA浓度。通过用该蛋白芯片检测50例CEA阳性HCC患者血清和56例健康人血清,对其进行盲法验证。结果:以CEA5 ng/m L为阳性判定标准,得出CEA蛋白芯片的灵敏度为92%(46/50),特异度为100%(56/56)。受试者工作特征(ROC)曲线分析显示该蛋白芯片检测出血清CEA的ROC曲线下面积(AUC)为0.960,与0.5相比差异有统计学意义(P0.001),其判定CEA阳性的准确性较高。结论:成功建立检测血清中CEA浓度的蛋白芯片,为下一步研发多种标志物联合检测HCC的蛋白芯片提供候选血清标志物。     

AFP-L3蛋白芯片的建立与应用

目的:建立一种可同时检测血清中甲胎蛋白(AFP)及甲胎蛋白异质体(AFP-L3)的蛋白芯片方法,为AFP-L3检测提供经济、便捷、省时、有效的新途径。方法:将鼠源AFP单克隆抗体和小扁豆凝集素点样固定在醛基玻片上,制备出AFP抗体和小扁豆凝集素蛋白芯片。利用抗原抗体特异性结合以及盐藻糖与小扁豆凝集素特异性结合的原理,用蛋白芯片方法检测血清样本中的AFP和AFP-L3。结果:肝癌血清39份,其中37份检测到AFP;26份肝癌血清中同时检测到AFP和AFP-L3,2份肝癌血清均未检测到AFP和AFP-L3。肝细胞癌组血清样本中AFP和AFP-L3水平明显高于健康对照组(P0.001),健康对照组与空白对照组统计学上无差异(P0.05)。结论:本研究成功地建立AFP和AFP-L3同时检测的蛋白芯片方法,与ELISA法和凝集素微量离心柱法相比,是一种切实可行,经济、便捷、省时、有效的方法。     

新型布尼亚病毒抗体检测方法的建立及初步应用

目的:建立新型布尼亚病毒IgG抗体ELISA检测方法。方法:用基因工程重组表达的新型布尼亚病毒NP抗原包被酶联板,建立间接ELISA法检测新型布尼亚病毒IgG抗体,并进行特异性和灵敏度评价,健康人群中检测结果计算临界值(均值+3标准差)。检测70例发热伴血小板减少综合征患者恢复血清和69份健康人血清样品。结果:在70份患者血清样品中,检测出新型布尼亚病毒IgG抗体阳性51例,阳性率为72.14%(51/70);69份健康人血清样品中,检测出1份阳性,特异性为98.6%(1/69)。结论:建立的新型布尼亚病毒IgG抗体ELISA检测方法特异性强、灵敏度高,可用于新型布尼亚病毒感染的检测及流行病学调查。     

抗SmD1抗体对系统性红斑狼疮诊断的临床意义

目的探讨抗SmD1抗体的测定对系统性红斑狼疮(SLE)诊断的临床意义。方法用免疫学方法分别检测SLE患者90例、非SLE病例对照组患者100例及健康对照组患者70例血清中的自身抗体。抗SmD1抗体的检测用ELISA方法;抗Sm抗体、抗双链DNA(dsDNA)抗体、抗核小体抗体(ANuA)的检测用免疫印迹法。结果 SLE组患者血清中抗SmD1抗体阳性率为75.6%;抗ANuA抗体阳性率为56.7%;抗Sm抗体的阳性率为24.4%;抗dsDNA抗体阳性率为47.8%。SmD1抗体阳性率明显高于ANuA抗体、Sm抗体及dsDNA抗体,差异有统计学意义(P0.05),且抗SmD1抗体的特异性为98.2%。结论与传统的检测指标比较,抗SmD1抗体具有较高的敏感性和特异性,可作为诊断SLE的一项重要的特异性检测指标。     

丙型肝炎病毒不同标志物检测结果的相关性分析

目的分析丙型肝炎病毒(Hepatitis C virus,HCV)抗原、抗体及核酸标志物实验室检测结果之间的关联性,评价适合血源筛查与早期临床确诊联合检测HCV感染的实验室诊断技术。方法用HCV RNA定量试剂、HCV核心抗原试剂及HCV抗体试剂分别检测304份血浆样本中HCV RNA载量、HCV Ag和抗-HCV指标,并对HCV RNA阳性样本进行基因分型。结果在304份血浆样本中,检出HCV RNA阳性样本87份,其HCV RNA载量≥500 IU/m L、500~30 IU/m L之间和<30 IU/m L时,血清学标志物HCV Ag浓度≥3 fmol/L及抗-HCV信号值/判断值≥1的阳性率分别为92.0%(23/25):96.0%(24/25)、58.8%(10/17):82.3%(14/17)及11.1%(5/45):75.6%(34/45),HCV RNA载量低于基因分型试剂盒LOD要求为500 IU/m L时,基因分型检测率为24.2%(15/62)。HCV RNA阴性样本217份中,HCV Ag浓度≥3 fmol/L和抗-HCV信号值/判断值≥1的样本阳性率分别为3.2%(7/217)和32.7%(71/217)。血清学指标在不同HCV RNA载量的阳性率差异具有统计学意义(χ2=197.4,P<0.01),HCV RNA载量与HCV Ag和抗-HCV水平成正相关。结论在中国人群中存在低HCV RNA水平携带者,HCV Ag和基因分型检测能力需要进一步提高。     

葡萄糖-6-磷酸异构酶抗原对类风湿关节炎的诊断价值

目的:通过比较类风湿关节炎(Rheumatoid arthritis,RA)患者、非RA风湿病患者及健康对照者血清中葡萄糖-6-磷酸异构酶(glucose-6-phosphate isomerase,GPI)抗原的阳性率来探讨GPI对RA的诊断意义,并探讨GPI,抗CCP抗体和RF联合应用对RA的诊断价值.方法:对110例RA患者、223例非RA风湿病患者和55例健康对照者共388份血清标本进行了检测.GPI抗原和抗CCP抗体采用ELISA方法,RF采用免疫比浊法定量检测.结果:RA组、非RA风湿病组和健康对照组血清中的GPI抗原阳性率分别为83.64%,30.04%和20.00%,RA组阳性率显著高于其它组(P＜0.01).GPI抗原对RAt诊断的敏感性和特异性分别为83.64%和71.58%.GPI和抗CCP抗体联合检测的敏感性和特异性分别为90.91%和71.22%,GPI和RF联合检测的敏感性和特异性分别为92.73%和 61.87%,如果三者同时检测,其敏感性和特异性分别为94.55%和60.43%.结论:RA患者的血清GPI阳性率明显高于其它自身免疫性疾病患者和健康对照者.GPI抗原时RA具有诊断价值,联合检测GPI、抗CCP抗体和RF可以提高RA诊断的敏感性.     

脱-γ-羧基凝血酶原的Ba-蛋白芯片法的建立和应用

目的:研发建立一种快速、特异性高、低成本的检测血清中脱-γ-羧基凝血酶原(DCP)的蛋白芯片,并通过检测DCP阳性肝细胞肝癌(HCC)血清临床样本对其进行评价。方法:首先采用柠檬酸钡吸附沉淀法去除血清中正常凝血酶原,然后通过双抗体夹心法蛋白芯片检测血清中DCP含量。通过Ba-蛋白芯片法检测41例DCP阳性HCC患者血清和66例健康人血清,进行盲法验证。结果:通过Ecarin-显色多肽底物法证实柠檬酸钡吸附沉淀法能够完全除血清中正常凝血酶原,根据"平均数+3×标准差"公式设置蛋白芯片的Cut-off值作为阳性判定标准,Ba-蛋白芯片法检测DCP阳性HCC血清的灵敏度为95.1%(39/41),特异度为100%(66/66)。结论:成功建立检测血清中DCP浓度的Ba-蛋白芯片法,为多种标志物联合检测HCC的蛋白芯片的研发提供候选血清标志物。     

广西地区野鼠戊型肝炎病毒感染情况调查

从广西壮族自治区5个市采集256份野鼠肝脏和对应的血清211份,采用HEV抗体(HEV-Ab)酶联免疫试剂盒(双抗原夹心法)对血清样本进行抗-HEV检测,并对不同地区HEV抗体阳性率进行χ2检验。采用HEV核酸荧光PCR试剂盒对肝脏样本进行HEV核酸检测。结果显示,211份野鼠血清抗-HEV的抗体阳性率为6.6%,其中百色市的抗体阳性率最高,为14.1%,其他地区抗体阳性率均在10%以下。256份野鼠肝脏中未检测到HEV核酸。由此可见,广西壮族自治区5个市的野鼠中存在HEV感染,但其是否为HEV的自然宿主及在戊型肝炎传播中起何作用需进一步研究。     

抗中性粒细胞胞浆抗体、抗核抗体联合检测对类风湿关节炎 的诊断价值

目的:探讨抗中性粒细胞胞浆抗体(ANCA)与抗核抗体(ANA)联合检测对类风湿关节炎的临床意义。方法:采用IIF法对82例RA患者(RA组)、74例非RA自身免疫疾病患者(非RA组)和52例健康体检者(正常对照组)的血清ANCA和ANA谱进行了检测分析,并用ELISA法进行抗丝氨酸蛋白酶3(PR3)、抗髓过氧化物酶(MPO)、ANA谱的定量检测。结果:RA组82例患者中,64例ANCA阳性,阳性率为78.08%,其中核周型(PANCA)37例,阳性率为45.1%,胞浆型(CANCA)27例,阳性率为32.9%;非RA组74例患者中有7例ANCA阳性率分别为9.4%;正常对照组50例中没有一例ANCA阳性。利用Elisa法对患者血清进行检测,分别能够特异的检测到PR3、MPO、抗双链DNA抗体(抗ds-DNA抗体)、抗ss-A等抗体、抗SS-A抗体、抗PM-SCL抗体的存在。结论:联合ANCA、ANA检测有助于提高类风湿关节炎的诊断。     

A Unique Protease Cleavage Site Predicted in the Spike Protein of the Novel Pneumonia Coronavirus (2019-nCoV) Potentially Related to Viral Transmissibility

正Dear Editor,In December 2019, a novel human coronavirus caused an epidemic of severe pneumonia(Coronavirus Disease 2019,COVID-19) in Wuhan, Hubei, China(Wu et al. 2020; Zhu et al. 2020). So far, this virus has spread to all areas of China and even to other countries. The epidemic has caused 67,102 confirmed infections with 1526 fatal cases     

Curcuminoids as inhibitors of thioredoxin reductase: A receptor based pharmacophore study with distance mapping of the active site

Curcumin is the yellow pigment of turmeric that interacts irreversibly forming an adduct with thioredoxin reductase (TrxR), an enzyme responsible for redox control of cell and defence against oxidative stress. Docking at both the active sites of TrxR was performed to compare the potency of three naturally occurring curcuminoids, namely curcumin, demethoxy curcumin and bis-demethoxy curcumin. Results show that active sites of TrxR occur at the junction of E and F chains. Volume and area of both cavities is predicted. It has been concluded by distance mapping of the most active conformations that Se atom of catalytic residue SeCYS498, is at a distance of 3.56 from C13 of demethoxy curcumin at the E chain active site, whereas C13 carbon atom forms adduct with Se atom of SeCys 498. We report that at least one methoxy group in curcuminoids is necessary for interation with catalytic residues of thioredoxin. Pharmacophore of both active sites of the TrxR receptor for curcumin and demethoxy curcumin molecules has been drawn and proposed for design and synthesis of most probable potent antiproliferative synthetic drugs.     

The structure, reproduction and taxonomy of Vidalia and Osmundaria (Rhodophyta, Rhodomelaceae)

Comprises species occurring mostly in subtidal habitats in tropical, subtropical and warm-temperate areas of the world. An analysis of the type species, V. spiralis (Sonder) Lamouroux ex J. Agardh, a species from Australia, establishes basic characters for distinguishing species in the genus. These characters are (1) branching patterns of thalli, (2) flat blades that may be spiralled on their axis, (3) width of the blade, (4) primary or secondary derivation of sterile and fertile branchlets and (5) position of sterile and fertile branchlets on the thalli. Application of the latter two characters provides an important basic method for separation of species into three major groups. Osmundaria , a genus known only in southern Australia, was studied in relation to Vidalia , and its separation from the Vidalia assemblage is not accepted. Species of Vidalia therefore are transferred to the older genus name, Osmundaria. Two new species, Osmundaria papenfussii and Osmundaria oliveae are described from Natal. Confusion in the usage of the epithet, Vidalia fimbriala Brown ex Turner has been clarified, and Vidalia gregaria Falkenberg, described as an epiphyte on Osmundaria pro/ifera Lamouroux, is revealed to be young branches of the host, Osmundaria prolifera.     

New or rare chromosome counts in Artemisia L. (Asteraceae, Anthemideae) and related genera from Kazakhstan

Fifteen chromosome counts of six Artemisia taxa and one species of each of the genera Brachanthemum, Hippolytia, Kaschgaria, Lepidolopsis and Turaniphytum are reported from Kazakhstan. Three of them are new reports, two are not consistent with previous counts and the remainder are confirmations of very scarce (one to four) earlier records. All the populations studied have the same basic chromosome number, x = 9, with ploidy levels ranging from 2x to 6x. Some correlations between ploidy level, morphological characters and distribution are noted.     

肝癌中HBV和HCV基因和抗原的分布及意义

采用原位分子杂交方法检测HCV RNA及HBV X基因;采用免疫组织化学方法研究HCV核心抗原,非结构区C33c抗原及HBxAg在肝细胞肝癌中的定位及分布.结果表明(1)HCV RNA、HBV X基因在肝细胞肝癌组织检出率分别为40%(55/136)和82%(112/136).HCV RNA定位于癌细胞的胞浆内,阳性细胞呈散在、灶状及弥漫分布三种形式;HBV X基因在肝癌细胞中的分布呈胞浆型、核型及核浆型,阳性细胞也呈上述三种分布形式;(2)HCV C33c抗原、核心抗原在肝细胞肝癌中的阳性率为81%(133/164)及86%(141/164).C33c抗原定位于癌细胞及肝细胞的胞浆内;核心抗原既定位于癌细胞核中,又可定位于胞浆中.C33c抗原阳性细胞以灶状分布为主;而核心抗原阳性细     

Comparative morphology of the carpel in the Liliaceae: Hewardieae, Petrosavieae, and Tricyrteae

The young pistils in the melanthioid tribes, Hewardieae, Petrosavieae and Tricyrteae, are uniformly tricarpellate and syncarpous. They lack raphide idioblasts. All are multiovulate, with bitegmic ovules. The Petrosavieae are marked by the presence of septal glands and incomplete syncarpy. Tepals and stamens adhere to the ovary in the Hewardieae and the Petrosavieae but not in the Tricyrteae. Two vascular bundles occur in the stamens of the Hewartlieae and Tricyrtis latifolia. Ventral bundles in the upper part of the ovary of the Hewardieae are continuous with compound septal bundles and placental bundles in the lower part. Putative ventral bundles occur in the alternate position in the Tricyrteae and putative placental bundles in the opposite. position in the Petrosavieae. The dichtomously branched stigma in each carpel of the Tricyrteae is supplied by a bifurcated dorsal bundle.