从氧自由基、一氧化氮探讨四逆汤抗急性失血性休克的肝脏机制

目的:从氧自由基、一氧化氮探讨四逆汤抗急性失血性休克的肝脏机制。方法:复制急性失血性休克大鼠模型, 分为假手术对照组;单纯休克模型组;休克+生理盐水复苏组;休克+四逆注射液复苏组。四逆注射液(浓度1000g生药/L), 剂量0.1mL/200g大鼠。用生理盐水或四逆注射液治疗3h后处死动物并取组织。测定各组肝脏超氧化物歧化酶(SOD)活性、丙二醛(MDA)水平, 一氧化氮(NO)水平。采用免疫组化染色法, 观察诱导型一氧化氮合酶(iNOS)在肝细胞中的变化特点。RT-PCR观测肝细胞iNOS和内皮源性一氧化氮合酶(eNOS)基因表达的变化。结果:模型组在休克1h后SOD活性明显低于对照组(P＜0.01)、MDA水平明显高于对照组(P＜0.01)。四逆汤组复苏3h后肝组织SOD明显高于生理盐水组(P＜0.01)、MDA低于生理盐水组(P＜0.01)、NO水平明显高于生理盐水组(P＜0.01)。生理盐水组iNOS在肝细胞中染色阳性单位明显高于四逆汤组(P＜0.05)。生理盐水组促进iNOSmRNA的表达。四逆汤组eNOSmRNA的表达增强。结论:四逆汤通过清除氧自由基, 升高NO, 改善肝组织微循环, 减少诱导型iNOS表达的各种因素, 理论上减轻了NO与氧自由基生成的ONOO的细胞毒作用和血管的低反应性, 并对肝脏起到保护作用。     

3'-Terminal RNA secondary structures are important for accumulation of tomato bushy stunt virus DI RNAs

The plus-strand RNA genome of tomato bushy stunt virus (TBSV) contains a 351-nucleotide (nt)-long 3'-untranslated region. We investigated the role of the 3'-proximal 130 nt of this sequence in viral RNA accumulation within the context of a TBSV defective interfering (DI) RNA. Sequence comparisons between different tombusviruses revealed that the 3' portion of the 130-nt sequence is highly conserved and deletion analysis confirmed that this segment is required for accumulation of DI RNAs in protoplasts. Computer-aided sequence analysis and in vitro solution structure probing indicated that the conserved sequence consists of three stem-loop (SL) structures (5'-SL3-SL2-SL1-3'). The existence of SLs 1 and 3 was also supported by comparative secondary structure analysis of sequenced tombusvirus genomes. Formation of the stem regions in all three SLs was found to be very important, and modification of the terminal loop sequences of SL1 and SL2, but not SL3, decreased DI RNA accumulation in vivo. For SL3, alterations to an internal loop resulted in significantly reduced DI RNA levels. Collectively, these data indicate that all three SLs are functionally relevant and contribute substantially to DI RNA accumulation. In addition, secondary structure analysis of other tombusvirus replicons and related virus genera revealed that a TBSV satellite RNA and members of the closely related genus Aureusvirus (family Tombusviridae) share fundamental elements of this general structural arrangement. Thus, this secondary structure model appears to extend beyond tombusvirus genomes. These conserved 3'-terminal RNA elements likely function in vivo by promoting and/or regulating minus-strand synthesis.     

Use of egg yolk-derived immunoglobulin as an alternative to antibiotic treatment for control of Helicobacter pylori infection

The present study evaluated the potential use of immunoglobulin prepared from the egg yolk of hens immunized with Helicobacter pylori (immunoglobulin Y [IgY]-Hp) in the treatment of H. pylori infections. The purity of our purified IgY-Hp was 91.3%, with a yield of 9.4 mg of IgY per ml of egg yolk. The titer for IgY-Hp was 16 times higher than that for IgY in egg yolk from nonimmunized hens, and IgY-Hp significantly inhibited the growth and urease activity of H. pylori in vitro. Bacterial adhesion on AGS cells was definitely reduced by preincubation of both H. pylori (10(8) CFU/ml) and 10 mg of IgY-Hp/ml. In Mongolian gerbil models, IgY-Hp decreased H. pylori-induced gastric mucosal injury as determined by the degree of lymphocyte and neutrophil infiltration. Therefore, in this experimental model, H. pylori-associated gastritis could be successfully treated by orally administered IgY-Hp. The immunological activity of IgY-Hp stayed active at 60 degrees C for 10 min, suggesting that pasteurization can be applied to sterilize the product. Fortification of food products with this immunoglobulin would significantly decrease the H. pylori infection. In conclusion, the IgY-Hp obtained from hens immunized by H. pylori could provide a novel alternative approach to treatment of H. pylori infection.     

Detection of factor Ⅷ intron 1 inversion in severe haemophilia A

Objective Screening the intron 1 inversion of factor Ⅷ (FⅧ) in the population of severe haemophilia A(HA) in China and performing carrier detection and prenatal diagnosis. Methods Using LD-PCR to detect intron 22 inversions and multiple-PCR within two tubes to intron 1 inversions in sereve HA patients. Carrier detection and prenatal diagnosis were performed in affected families. Linkage analysis and DNA sequencing were used to verify these tests. Results One hundred and eighteen patients were seven diagnosed as intron 22 inversions and 7 were intron 1 inversions out of 247 severe HA patients. The prevalence of the intron 1 inversion in Chinese severe haemophilia A patients was 2. 8% (7/247). Six women from family A and 2 from family B were diagnosed as carriers. One fetus from family A was affected fetus. Conclusion Intron 1 inversion could be detected directly by multiple-PCR within two tubes. This method made the strategy more perfective in carrier and prenatal diagnosis of haemophilia A.     

中国恒河猴(Macaca mulatta)外周血CD4+CD25+T淋巴细胞的研究

目的:研究中国恒河猴外周血中CD4 CD25 T淋巴细胞亚群及其分布频率。方法:利用流式细胞术对50只中国恒河猴外周血CD4 CD25 T淋巴细胞进行了分析。结果:发现所有被检测的恒河猴个体中均存在明显的CD4 CD25 T淋巴细胞亚群;CD4 CD25 T淋巴细胞大约占CD4 T淋巴细胞的9.1%(变化范围为2.6%～18.1%);其中CD4 CD25highT淋巴细胞约占2.5%(0.3%～5.5%)。对不同年龄和性别个体中CD4 CD25 T淋巴细胞频率的初步分析未发现统计学上有年龄或性别差异。结论:中国恒河猴可用于与CD4 CD25 T细胞相关的人类疾病的研究中。     

Development and selection of γδ T cells

Summary:  Two main lineages of T cells develop in the thymus: those that express the αβ T-cell receptor (TCR) and those that express the γδ TCR. Whereas the development, selection, and peripheral localization of newly differentiated αβ T cells are understood in some detail, these processes are less well characterized in γδ T cells. This review describes research carried out in this laboratory and others, which addresses several key aspects of γδ T-cell development, including the decision of precursor cells to differentiate into the γδ versus αβ lineage, the ordered differentiation over the course of ontogeny of functional γδ T-cell subsets expressing distinct TCR structures, programming of ordered Vγ gene rearrangement in the thymus, including a molecular switch that ensures appropriate Vγ rearrangements at the appropriate stage of development, positive selection in the thymus of γδ T cells destined for the epidermis, and the acquisition by developing γδ T cells of cues that determine their correct localization in the periphery. This research suggests a coordination of molecularly programmed events and cellular selection, which enables specialization of the thymus for production of distinct T-cell subsets at different stages of development.     

小发卡RNA抗呼吸道合胞病毒效应的研究

目的 为从基因水平抑制呼吸道合胞病毒（respiratory syncytial virus，RSV）的复制，针对RSV的M2-2基因构建小发卡结构状RNA（short hairpin RNA，shRNA）重组质粒，在细胞水平观察shRNA对RSV复制的影响。方法 将成功构建的针对RSV M2-2基因的重组质粒pshRNA8260转染HEp-2细胞，利用光镜观察pshRNA8260对RSV致HEp-2细胞病变效应（cytopathogenic effect，CPE）的影响并计算CPE抑制率，空斑形成实验检测RSV滴度变化。结果 成功构建了针对人RSV M2-2基因mRNA的pshRNA8260重组质粒，研究发现pshRNA8260能明显改善RSV所致的病变效应，降低RSV在细胞内复制的病毒滴度。结论 针对RSV M2-2基因的pshRNA82（g）重组质粒具有明显的特异性抗RSV效应的作用。     

中小学校学生流感疫苗接种效果评估和卫生经济学评价

目的评估中小学校学生流感疫苗接种效果,开展疫苗接种的卫生经济学评价,为制定流感疫苗接种策略提供依据。方法采用易感者-潜伏者-显性感染者/隐性感染者-移出者/恢复者(susceptible-exposed-infectious/asymptomatic-removed/recovered,SEIARR)动力学模型对既往疫情数据进行模拟,计算累计罹患率评估疫苗接种效果,计算成本-效果和成本-效益进行卫生经济学评价。结果当流感疫苗接种率为0%、50%、70%和90%时,疫情平均累积罹患率为99.79%(99.75%,99.82%)、70.84%(70.58%,71.09%)、56.81%(56.17%,57.45%)和34.29%(32.46%,36.20%),流感疫苗接种率越高,累计罹患率越低;当疫苗接种率为50%、70%和90%时,疫苗的成本-效果比为:141.03(139.86,142.19)元、133.16(131.34,135.08)元和112.90(110.03,116.11)元,疫苗的成本-效益比为:6.27(6.22,6.32)元、6.65(6.56,6.74)元和7.88(7.66,8.09)元;流感疫苗接种率越高,疫苗的成本-效果越好,成本-效益越高。结论接种流感疫苗能有效降低疫情的累计罹患率,提高接种率能有效增加疫苗接种的卫生经济学效应。     

遗尿儿童和青少年情绪和行为问题及相关因素研究

目的 调查河南省儿童青少年遗尿症的情绪和行为问题的现况及其影响因素。 方法 从2019年10月—2020年3月,采用分层随机整群抽样方法抽取河南某县5所中小学校,采用一般资料调查表、长处和困难问卷及儿童睡眠调查表进行调查,对影响遗尿患儿情绪和行为的因素进行单因素、多因素分析。 结果 实际发放问卷4 500份,回收4 120份,有效问卷3 912份,有效回收率为94.95%。调查结果显示有7.82%儿童患有遗尿症。遗尿组的情绪症状、多动和困难总分得分明显高于正常组(P＜0.001),两组品行问题、同伴交往和社会行为得分的比较差异无统计学意义(P＞0.05)。Pearson相关分析结果显示,遗尿儿童和青少年的困难总分与打鼾、夜间呼吸问题、嗜睡、注意力缺陷、睡眠总分均存在正相关关系(P＜0.05)。多元线性回归结果显示睡眠呼吸紊乱、照顾者文化程度低、便秘、学习成绩较差是遗尿患儿情绪和行为问题的影响因素。 结论 遗尿患儿情绪和行为问题的检出率高,影响因素是睡眠呼吸紊乱、照顾者文化程度低、孩子学习成绩差、便秘,应引起关注。     

2010—2020年云南省学校食源性疾病暴发事件流行病学分析

目的 了解云南省学校食源性疾病暴发事件流行病学特征和趋势。 方法 收集2010—2020年云南省各地报告的学校食源性疾病暴发资料,并进行描述性流行病学分析。 结果 2010—2020年共发生学校食源性疾病暴发260起,发病6 600人,死亡2人,总体呈下降趋势。学校食源性疾病暴发高峰期为第二、四季度,占总起数的66.54%(173/260);暴发场所多发生在小学,占总起数的58.08%(151/260)。报告明确或可疑致病因子的185起事件中由微生物或可疑微生物引起的占40.00%(74/185),毒素引起的占46.49%(86/185);明确致病因子的118起暴发事件中,60.17%(71/118)是由4种致病因子引起的,其中蓖麻毒素导致的最多,达22起(18.64%),发病人数428人,其次是蜡样芽孢杆菌20起(16.95%),原因食品被归因为一类食品植物类,主要是大米、三明治、面包。导致暴发污染环节最多的是生产加工环节(50.77%,132/260)。 结论 关注学校食源性疾病暴发事件流行特征,在学生开学季时,加强对小学食堂植物类食品的生产加工环节管理和监督。微生物、毒素感染是学校食源性疾病主要的致病因素,且原因查明率低,进一步强化医疗机构食源性疾病微生物、毒素检测能力,提高原因查明率,最大限度地降低学校食源性疾病的发病率。