IFN-γ与 IL-4基因多态性与儿童哮喘易感性及外周血IFN-γ、IL-4和IgE的关系

目的: 探讨IFN-γ和 IL-4 基因多态性与儿童哮喘易感性及血浆IFN-γ、IL-4和IgE的相关性。方法: 用聚合酶链反应-限制性酶切片段长度多态性(PCR-RFLP)法检测100例哮喘儿童和122例对照儿童IFN-γ基因-179G/T、 IL-4 基因-33C/T和-589C/T位点基因型;等位基因特异性-聚合酶链反应(AS-PCR)法检测IFN-γ基因+874A/T位点基因型;毛细管电泳法检测IFN-γ基因CA重复序列基因型;ELISA法测定血浆IFN-γ、IL-4和IgE。结果: 100例哮喘儿童和122例对照儿童IFN-γ基因-179位点均为GG纯合子,未检测到突变基因型。IFN-γ基因+874A/T位点和CA重复序列的基因型和等位基因频率分布在哮喘组和对照组间无显著差异(P>0.05);+874位点多态性与血浆IFN-γ水平相关,AA基因型IFN-γ含量低于AT基因型(P<0.05)。 IL-4 基因-33C/T和-589C/T位点的基因型和等位基因频率分布在哮喘组和对照组间有显著差异(P<0.05);-33和-589位点TT基因型外周血IL-4和总IgE浓度均高于CT基因型,只有-33位点与血浆IL-4水平存在相关性(P<0.05)。结论: IFN-γ基因+874A/T和CA重复序列多态性可能与儿童哮喘无相关性,+874A/T位点多态性与IFN-γ水平相关。 IL-4 基因-33TT和-589TT基因型可能为儿童哮喘的易感基因型,-33位点多态性与IL-4表达水平相关。     

云南汉族人群IL-4基因5’和3’非编码区域多态性与HCV慢性感染无相关性

目的探讨IL-4基因5’和3’非编码区域单核苷酸多态性位点(single nucleotide polymorphism,SNP)与HCV慢性感染的相关性。方法选取云南地区汉族人群HCV慢性感染患者380例,健康体检人群439例。采用Taq Man探针基因分型方法对IL-4基因5’和3’非编码区域6个SNP位点SNP-1138A/G(rs2243247)、-1098G/T(rs2243248)、-589C/T(rs2243250)、-33C/T(rs2070874)、2979C/A(rs2227284)、3’端C/T(rs2243292)进行基因分型,并构建单倍型,评估上述6个SNP位点及单倍型与HCV慢性感染的相关性。结果 IL-4基因SNP-1138A/G(rs2243247),3’端C/T(rs2243292)在病例组和对照组中无多态性;SNP位点-1098G/T(rs2243248)、-589C/T(rs2243250)、-33C/T(rs2070874)、2979C/A(rs2227284)的基因型频率和等位基因频率在病例组和对照组中差异无统计学意义(P0.05);单倍型分析结果显示:SNP位点-1098G/T(rs2243248)、-589C/T(rs2243250)、-33C/T(rs2070874)、2979C/A(rs2227284)构建的单倍型频率在病例组和对照组中差异无统计学意义(P0.05)。结论在云南汉族群体中,IL-4基因5’和3’非编码区域SNP位点-1138A/G(rs2243247)、-1098G/T(rs2243248)、-589C/T(rs2243250)、-33C/T(rs2070874)、2979C/A(rs2227284)、3’端C/T(rs2243292)与HCV慢性感染没有相关性。     

白介素-10-627基因多态性和儿童哮喘的相关性研究

目的探讨白介素-10(IL-10)基因-627位点多态性与昆明地区儿童哮喘的相关性。方法采用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)分析方法,检测50例哮喘儿童和36例健康儿童IL-10基因-627位点基因型,比较两组IL-10基因-627位点的基因型和等位基因分布频率。结果哮喘组和健康组3种基因型CC、CA、AA分别为:12.0%、48.0%、40.0%;8.3%、50.0%、41.7%。两组基因型分布差异无显著性(P>0.05)。哮喘组和健康组C和A的等位基因分布频率分别为:36.0%、64.0%;31.9%、68.1%。两组等位基因分布频率差异无显著性(P>0.05)。结论本研究表明IL-10基因-627位点多态性可能与昆明地区儿童哮喘易感性无关。     

IL-4R、IL-13、ADAM33基因多态性与中国皖南地区汉族人群支气管哮喘的相关性研究

目的:探讨IL-4受体基因Arg551Gln(rs1801275)、IL-13基因Arg130Gln(rs20541)、ADAM33基因T1(rs2280091)位点基因多态性与中国皖南地区汉族人群支气管哮喘的相关性。方法:采用病例-对照的方法,用聚合酶链反应及直接基因测序法比较116例支气管哮喘组与70例正常人对照组之间基因型、等位基因频率的差异。结果:哮喘组和对照组IL-4受体基因Arg551Gln位点和IL-13基因Arg130Gln位点的基因型和等位基因型频率的差异有统计学意义,ADAM33基因T1位点基因型哮喘组和对照组差异有统计学意义,等位基因型频率在哮喘组和对照组差异无统计学意义。结论:提示IL-4R Arg551Gln(rs1801275)位和IL-13基因Arg130Gln(rs20541)位的多态性可能与中国皖南地区汉族哮喘有相关性;ADAM33基因(rs2280091)T1位点位的多态性可能与中国皖南地区汉族哮喘无相关性。     

白介素10的基因多态性与尖锐湿疣的相关性研究

目的探讨白介素10（IL-10）基因启动子-1082位点基因多态性与尖锐湿疣的相关性。方法采用焦磷酸测序法（Pyrosequencing）检测30例尖锐湿疣患者（观察组）和50例健康体检者（对照组）IL-10基因启动子-1082G/A位点基因型和等位基因频率;同时采用双抗体夹心ELISA法测定对照组和观察组的血清IL-10水平。结果观察组血清IL-10水平显著高于对照组（P〈0.01）。观察组IL-10基因启动子-1082 G/A位点GG基因型分布频率和G等位基因频率高于对照组（P〈0.01）。在观察组中表达GG基因型患者的血清IL-10水平显著高于表达其它基因型患者的血清IL-10水平（P〈0.05）。结论 IL-10基因多态性与尖锐湿疣易感性可能相关,IL-10基因启动子-1082 G/A位点GG基因型携带者对尖锐湿疣的易感性高。     

rhGM-CSF作为成人乙肝疫苗无(弱)应答者免疫佐剂初探

目的 探讨重组人粒细胞.巨噬细胞集落刺激因子(rhGM-CSF)对提高成人乙肝疫苗无(弱)应答免疫的作用.方法 将两年内完成1～2个标准乙肝疫苗接种程序、复查HBV标志物均阴性的健康人群随机分为A、B两组.A组:33人,按标准免疫程序予10μg乙肝疫苗;B组:34人,先予rhGM-CSF 300μg皮下注射,次日始按A组方案予乙肝疫苗.接种首针后第1、2、8个月(T1、T2、T8)采血检测抗-HBs.结果 A、B组T8抗-HBs阳性率分别为39.39%和64.71%(P=0.038);A组三次抗体滴度检测结果 无显著性变化;B组升高明显,分别为(113.85±198.56)mIU/ml,(312.40±349.44)mIU/ml,(427.74±411.58)mIU/ml(P=0.001).A组和B组T8抗体水平差异有统计学意义(P=0.010).结论 rhGM-CSF联合乙肝疫苗复种对无(弱)应答者免疫效果优于单纯复种,GM-CSF具有提高机体对乙肝疫苗免疫应答的能力.     

新生儿乙肝酵母疫苗免疫后抗体无应答者的再免、加强免疫与细胞免疫的研究

目的 HBsAg阴性母亲婴儿应用国产乙型肝炎重组酵母疫苗常规接种后的抗-HBs无应答者的再免、加强免疫效果,以及抗体阴性婴儿细胞免疫应答状况.方法 在河南省开封市筛选8093例7～24月龄,母亲HBsAg阴性,按0、1、6月程序接种国产5 μg乙肝酵母疫苗的健康婴儿,RIA法检测抗-HBs.对无应答者进行5 μg×2或×3;10 μg×2或×3剂4组疫苗再免、加强免疫效果的比较.随机选择加免后抗体阳性和仍阴性的婴儿,用ELISPOT方法测定PBMC体外刺激产生的IFN-r和IL-2.结果 2970例7～10月龄婴儿中56例抗-HBs低于10 mIU/ml者分4组再免后,抗-HBs阳转率＞85%.而11～24月龄5933例中346例无应答者也同上分为4组.加强免疫后,抗-HBs阳转率为92.59%～97.92%,无统计学差异.10例加强免疫后抗体仍持续阴性婴儿TH1类细胞因子IL-2和IFN-r的阳性率分别为30%和20%,低于有应答人群(19/32,59.38%;11/32,37.04%).结论 母亲HBsAg阴性的婴儿常规接种5 μg乙肝疫苗后抗体无应答者,用5 μg、10 μg两剂或三剂加强免疫都有良好的抗体阳转效果.加免抗体仍阴性婴儿与正常应答者存在明显的细胞免疫应答的差别.     

肿瘤坏死因子-α基因启动子多态性与慢性乙型肝炎易感性的关系

目的探讨湖北汉族人肿瘤坏死因子-α(TNF-α)基因启动子多态性及其与慢性乙型肝炎易感性之间的关系。方法应用聚合酶链反应-限制性片段长度多态性分析方法(RFLP),检测126名正常对照者和131例慢性乙型肝炎患者TNF-α基因启动子多态性。结果共发现12种启动子基因型,以GG.GG.CC.CC、GG.GG.CC.CA、GG.GG.CT.CC和GG.GA.CC.CC基因型多见,约占85%;通过对TNF-α基因启动子4个位点基因型分析发现,慢性乙型肝炎患者和正常对照者TNF-α基因启动子-238G/A、-857C/T位点基因型分布频率差异无显著性,而-308G/A、-863C/A位点基因型分布频率差异有显著性。结论湖北汉族人慢性乙型肝炎易感性与TNF-α基因启动子-308G/A、-863C/A位点多态性有关,其中TNF-α-308GA、-863CA基因型携带者患慢性乙型肝炎风险相对较小。     

汉族人白细胞介素-18基因启动子多态性与慢性乙型肝炎的相关性研究

目的 探讨中国汉族人白细胞介素-18（interleukin-18,IL-18）基因启动子单核苷酸多态性及其与慢性乙型肝炎易感性之间的关系。方法 应用序列特异性引物一聚合酶链反应技术，检测231例慢性乙型肝炎患者和300名正常人儿．馏基因启动子-607C/A、-137G／C单核苷酸多态性位点基因型。结果 正常对照组和慢性乙型肝炎组中，IL-18基因启动子-607C/A位点3种基因型频率分别为CC型：0．22（66／300）和0．27（62／231），CA型：0．53（160／300）和0．50（116／231），AA型：0．25（74／300）和0．23（53／231）；IL-18基因启动子-137G／C位点3种基因型频率分别为GG型：0．67（202／300）和0．79（182／231），GC型：0．30（90／300）和0．19（45／231），CC型：0．03（8／300）和0．02（4／231）。经Y0检验，慢性乙型肝炎组IL-18基因启动子-137GG分布频率显著高于正常对照组（X^2=8．55，P=0．003），而-607C／-137C和-607A／-137C单倍型频率显著低于正常对照组。进一步比较慢性乙型肝炎患者儿．馏基因启动子多态性与乙型肝炎病毒（hepatitis Bvirus，HBV）DNA复制的关系，发现高水平HBV—DNA组-607位点AA基因型分布频率明显低于低水平HBV—DNA组（Y2=6．03，P=0．014）。结论 汉族人慢性乙型肝炎与IL-18基因启动子-607C/A、-137G／C单核苷酸多态性相关，其中IL-18基因启动子-137位点C等位基因可能对机体HBV感染有保护作用，而启动子-607位点AA型对感染后HBV—DNA的复制可能有抑制作用。     

IL-17、IL-33水平及IL-33基因rs16924159G/A位点多态性与不明原因复发性流产易感性分析

目的了解深圳地区不明原因复发性流产(unexplained recurrent spontaneous abortion,URSA)患者血清中白细胞介素-17(interleukin-17,IL-17)、IL-33水平及IL-33基因rs16924159G/A位点多态性分布情况,并探讨其与URSA发病的易感性。方法收集2019年2月～2020年1月来医院妇科门诊就诊并确诊为URSA患者132例为URSA组,并选择同期来医院妇科门诊要求人工流产的早孕女性120名为对照组,采用酶联免疫吸取附法(ELISA)检测血清中IL-17和IL-33水平,同时采用多重连接酶检测反应(improved multi-ligase detection reaction,imLDR)技术检测IL-33基因rs16924159G/A位点多态性。结果 URSA患者血清中IL-17水平为56.01±19.43ng/L,明显高于对照组的32.85±11.27ng/L,而IL-33水平为105.97±26.32pg/ml,明显低于对照组的154.68±34.76pg/ml,差异均有统计学意义(P0.05);URSA患者IL-33基因rs16924159G/A位点AA基因型和A等位基因检出率分别为28.03%和44.94%,明显高于对照组的9.17%和23.75%,差异均有统计学意义(P0.05);携带IL-33基因rs16924159G/A位点AA基因型URSA患者血清中IL-33水平为76.08±19.25pg/ml,明显低于GG和GA基因型的121.52±30.67pg/ml和113.71±28.06pg/ml,差异有统计学意义(P0.05),而GG和GA基因型之间差异无统计学意义(P0.05),同时对照组中不同基因型血清中IL-33水平之间差异均无统计学意义(P0.05);经Spearman相关性分析,URSA患者血清中IL-17和IL-33水平呈明显负相关(r=-0.6706,P0.05)。结论 URSA发病可能与IL-33水平降低,其对Th17细胞分泌抑制作用减弱有关,而IL-33水平降低可能与其基因rs16924159G/A位点多态性突变有关,其中AA基因型可能是URSA发病的危险易感基因之一。     

Development in in vitro toxicology

There are three principal pressures driving the development of in vitro toxicology: (1) the need for more efficient testing systems to cope with the large number of xenobiotics currently being developed; (2) public pressure to reduce animal experimentation; and (3) a need for a better understanding of the mechanisms of toxicity. Within this, in vitro toxicology is focused on local, systemic, and target-organ toxicity. It is becoming increasingly apparent that a step or decision-tree approach using input of a variety of experimental data (physicochemical properties, biokinetics, cytotoxicity) provides the most efficient system for predicting toxicity. Examples of the use of in vitro toxicity systems for prediction of systemic toxicity and target-organ (liver) toxicity are presented.Originally presented at ECCP 93.     

Seasonal variations in acute toxoplasmosis in pregnant women in Slovenia

Between December 1999 and December 2004, 40 081 pregnant women were examined for toxoplasmosis with Toxo-IgG, Toxo-IgM enzyme immunoassay. Women with positive results were then retested with the Toxo-IgG avidity assay for recent toxoplasmosis. Recent acute toxoplasmosis in pregnant women was found to be significantly more frequent (p < 0.01) during winter than summer. The incidence of acute toxoplasmosis during winter-spring was also significantly more frequent (p < 0.025) than summer-autumn. This phenomenon should be taken into account when formulating preventive measures for toxoplasmosis, especially for pregnant women.     

Age-related changes in polyamines in memory-associated brain structures in rats

Polyamines putrescine, spermidine and spermine are positively charged aliphatic amines and have important roles in maintaining normal cellular function, regulating neurotransmitter receptors and modulating learning and memory. Recent evidence suggests a role of putrescine in hippocampal neurogenesis, that is significantly impaired during aging. The present study measured the polyamine levels in memory-related brain structures in 24- (aged), 12- (middle-aged) and 4- (young) month-old rats using liquid chromatography/mass spectrometry and high performance liquid chromatography. In the hippocampus, the putrescine levels were significantly decreased in the CA1 and dentate gyrus, and increased in the CA2/3 with age. Significant age-related increases in the spermidine levels were found in the CA1 and CA2/3. There was no difference between groups in spermine in any sub-regions examined. In the parahippocampal region, increased putrescine level with age was observed in the entorhinal cortex, and age did not alter the spermidine levels. The spermine level was significantly decreased in the perirhinal cortex and increased in the postrhinal cortex with age. In the prefrontal cortex, there was age-related decrease in putrescine, and the spermidine and spermine levels were significantly increased with age. This study, for the first time, demonstrates age-related region-specific changes in polyamines in memory-associated structures, suggesting that polyamine system dysfunction may potentially contribute to aged-related impairments in hippocampal neurogenesis and learning and memory.     

休克过程中肾上腺髓质素变化的研究进展

Adrenomedullin (AM) is a new peptidergic regulator of vascular function. AM serves as a hormone, which has many biological properties, plays an important role in the many pathophysiological processes, especially shock. This review will highlight the structure, biological properties of AM and the relationship between AM and shock.     

Secular change in menarche in women in Madrid

The age at menarche was estimated by recollection in 1617 women between the ages of 18 and 60 in Madrid and a nearby suburb, Pinto. The population of Pinto is working-class and the Madrid group, taken from residential neighbourhoods , belongs to the upper middle class. In both groups we found a diminution in average age at menarche, from 14.04 to 13.02 years in Madrid and from 14.55 to 13.16 years from about 1935 to about 1965 in Pinto. These changes have been more intense in the group which is less well-off economically, where living conditions have varied much more drastically.     

Pitfalls in TRAP assay in routine detection of malignancy in effusions

Telomerase has been found to be reactivated in a majority of cancers but is inactive in most somatic cells. Our principal goal was to determine the potential use of the telomeric repeat amplification protocol (TRAP) assay as marker for malignancy in cytological effusions. The simple selection criterion was the cytological diagnosis, and routine samples were classified into malignant (58 samples) and nonmalignant (233 samples). Of the malignant samples, 44/58 (76%) were positive by TRAP assay. Of the 14 telomerase-negative cytology-positive samples, RNA integrity was poor in 9, indicating suboptimal sample conservation for molecular analysis. In 3 of the remaining 5 samples with a negative TRAP assay, a high number of malignant cells was observed, and these cells might have been telomerase-negative. Thus, the sensitivity of TRAP assay for the presence of malignant cells was about 76%. In the cytologically nonmalignant effusions, the presence of telomerase activity was observed in 24% (55/233). Of these, 6% were highly suspicious for malignancy, 9% were doubtful, and 9% were cytologically nonmalignant effusions confirmed by a follow-up of 12 mo or more. According to these data, the specificity of the TRAP assay to detect tumor cells in effusions ranged only between 82-91%. Our results indicate that, although the TRAP assay is positive in 6-15% of putative malignant effusions, the relatively high number of TRAP false-negative and false-positive cases renders this test unsuitable for routine diagnostic purposes.