全基因组测序确诊氨基甲酰磷酸合成酶缺乏症Ⅰ型1例

<正>患儿，男，1个月20天，因“呕吐3天，昏睡、呻吟半天伴反复抽搐发作”入院，患儿系G2P2,足月顺产，否认出生时有窒息抢救史，出生体重3 400 g, 父母体健，否认近亲结婚，否认家族遗传病、传染病及肝胆疾病史。患儿哥哥体健。入院查体：昏睡状态，精神、反应差，面色稍苍白，无特殊面容，全身皮肤无黄染、淤点淤斑、出血点及花纹征，呼吸深快，三凹征(-),心肺听诊阴性，腹膨隆、软，腹壁未见静脉曲张，肝脾肋下未及，肠鸣音正常，四肢肌张力增高，     

应用多重荧光Real-time PCR快速鉴定猪链球菌2型

目的建立能同时鉴定链球菌2型(S.suis2)和确定主要毒力标志MRP的快速检测方法。方法根据S.suis2CPS2J与MRP的保守序列,设计并合成了2对引物及其相应探针。通过优化各反应物的浓度、配比和循环程序,建立能从组织中直接检测CPS2J和MRP的实时荧光PCR方法。结果能在3h内对送检组织样品检测确定S.suis2型和MRP毒力基因,最低检测菌体浓度24CFU/ml。与普通PCR相比,敏感性在检测CPS2J时比普通PCR高至少10倍以上,而在检测MRP时比其高100倍。与葡萄球菌、化脓链球菌、沙门氏菌、S.suis1型、9型等无交叉反应。同一人在试剂放置0d、20d和60d检测,以及不同人用同一方法检测,变异系数均小于5%,差异不显著。用于检测SSsc0501攻毒后的样品,结果在心、肝、脾、肺、肾、淋巴结、脑中均检测到S.suis2阳性,MRP阳性。而在肌肉中,S.suis2和MRP均为阴性。结论建立的多重荧光Real-timePCR敏感性高、特异性强,稳定性和重复性好,能用于可疑S.suis2型组织样品的快速定型和毒力鉴定。     

等离子体低温减容术治疗阻塞性睡眠呼吸暂停低通气综合征

等离子体低温减容术(CPS)是一种治疗阻塞性睡眠呼吸暂停低通气综合征(OSAHS)的新型微创技术。为了验证CPS治疗OSAHS的实际效果和适应证，2001年4月至2003年4月，我们对CPS治疗OSAHS进行了临床研究，现报告如下。     

CYP1A1 , GSTM1 , GSTT1 and NQO1 polymorphisms and colorectal adenomas in Japanese men

AIM: To investigate the role of functional genetic poly-morphisms of metabolic enzymes of tobacco carcinogens in the development of colorectal adenomas. METHODS: The study subjects were 455 patients with colorectal adenomas and 1052 controls with no polyps who underwent total colonoscopy in a preretirement health examination at two Self Defense Forces hospitals. The genetic polymorphisms studied wereCYP1A1*2A (rs 4646903), CYP1A1*2C (rs 1048943), GSTM1 (null or non-null genotype), GSTT1 (null or non-null genotype) and NQO1 C609T (rs 1800566). Genotypes were determined by the polymerase chain reaction (PCR)-restriction fragment length polymorphism or PCR method using genomic DNA extracted from the buffy coat. Cigarette smoking and other life-style factors were ascertained by a self-administered questionnaire. The associations of the polymorphisms with colorectal adenomas were examined by means of OR and 95%CI, which were derived from logistic regression analysis. Statistical adjustment was made for smoking, alcohol use, body mass index and other factors. The gene-gene interaction and effect modification of smoking were evaluated by the likelihood ratio test. RESULTS: None of the five polymorphisms showed a significant association with colorectal adenomas, nor was the combination of GSTM1 and GSTT1 . A borderline significant interaction was observed for the combination of CYP1A1*2C and NQO1 (P = 0.051). The OR associated with CYP1A1*2C was significantly lower than unity among individuals with the NQO1 609CC genotype. The adjusted OR for the combination of the CYP1A1*2C allele and NQO1 609CC genotype was 0.61 (95%CI: 0.42-0.91). Although the interaction was not statistically significant (P = 0.24), the OR for individuals carrying the CYP1A1*2C allele and GSTT1 null genotype decreased significantly compared with those who had neither CYP1A1*2C allele nor GSTT1 null genotype (adjusted OR: 0.69, 95%CI: 0.49-0.97). Smoking did not modify the associations of the individual polymorphisms with colorectal adenomas. There w     

1,1,1-三氯乙烷致急性中毒性肝炎19例

1,1,1-三氯乙烷(TCE)为工业用品,属低毒类物质。近年来由于其在工业上的应用,TCE的毒性及其对工人健康的危害引起人们的关注。我院2003年1月共收治泉州市某陶瓷厂车间因通风不良而致TCE急性中毒19例,结果报道如下。 1.临床资料:(1)病例:2003年1月收治泉州市某陶瓷厂车间集体发病的患者19例,其发病率为63.3％(19/30);男5例,女14例,年龄23～40岁,平均30.5岁。员工均在工厂食堂进     

胰高血糖素样肽-1抗动脉粥样硬化的效果和机制研究

目的 探索胰高血糖素样肽（GLP）-1对氧化低密度脂蛋白（ox-LDL）诱导的人脐静脉内皮细胞（HUVECs）损伤的作用及机制。方法 将体外培养HUVECs分为对照组（无处理）和GLP-1预处理组（分别给予0、2.5、5和10 nmol/L GLP-1预处理24 h后,再以100 μg/ml ox-LDL氧化24 h）。应用MTT法检测细胞生存率,评估GLP-1在ox-LDL诱导HUVECs损伤中的作用;应用免疫荧光技术评估GLP-1在ox-LDL诱导单核细胞对HUVECs的黏附的影响;应用ELISA法检测HUVECs在接受ox-LDL和GLP-1处理后E-选择素,细胞间黏附分子（ICAM）-1、血管细胞黏附分子（VCAM）-1的分泌,并应用RT-PCR法检测E-选择素、ICAM-1、VCAM-1基因表达。结果 MTT试验显示（2.5、5和10 μmol/L）的GLP-1均可降低ox-LDL对HUVECs的杀伤作用,保护作用与浓度呈正相关（P<0.05）;免疫荧光检测结果显示,GLP-1可以降低单核细胞对ox-LDL损伤HUVECs的黏附作用,且和浓度呈正相关（P<0.05）。GLP-1处理后的ICAM-1、VCAM-1及E-选择素的表达和分泌均有显著下降（均有P<0.05）。结论 GLP-1可能通过对抗HUVECs氧化损伤,下调ICAM-1、VCAM-1和E-选择素的表达与分泌,减少单核细胞趋化和黏附,发挥抗动脉粥样硬化作用。     

PD-1/PD-L1途径在慢性病毒感染中的作用

程序性死亡因子配体-1(PD-L1)分子是B7超家族的重要成员,在单核细胞、树突状细胞及大部分外周组织可以广泛诱导性表达。PD-L1可以和程序性死亡-1(PD-1)及CD80受体结合调节淋巴细胞的功能。PD-1/PD-L1信号在病毒感染后效应T细胞免疫耐受中具有重要作用,并且预示阻断该途径可能是抗病毒感染治疗的方向之一。     

PD1/PD-L1信号通路相关蛋白在肝癌中的表达及与临床病理特征的关系

目的探讨肝癌患者PD1/PD-L1信号通路相关蛋白的表达特点及与临床病理相关性。方法选取2015年1月-2018年12月内蒙古包钢医院就诊的80例肝癌患者作为研究对象。RT-PCR和Western blot检测PD1和PD-L1 mRNA和蛋白表达水平。免疫组化法检测组织中PD1和PD-L1的表达水平。结果肝癌组PD1和PD-L1 mRNA和蛋白水平高于对照组,差异有统计学意义(P0.05)。肝癌组PD1和PD-L1表达阳性率高于对照组,差异有统计学意义(P0.05)。统计分析证实PD1表达与肝癌患者肿瘤转移密切相关,差异均有统计学意义(P均0.05)。PD-L1表达与肝癌患者肿瘤直径密切相关,差异均有统计学意义(P均0.05)。结论 PD1和PD-L1在肝癌中表达显著增高,且可能与肝癌发生、发展及预后相关。     

Plk1在胰腺癌中的表达及临床意义

目的 研究胰腺癌组织标本及细胞系中Plk1的表达及临床意义。方法 应用免疫组化EnVision两步法检测33例胰腺癌及其癌旁组织、3例胰腺浆液性囊腺瘤、3例黏液性囊腺瘤、5例慢性胰腺炎的石蜡标本中Plk1的表达。用半定量RT—PCR检测5例新鲜胰腺癌组织及其癌旁组织中Plk1 mRNA的表达。用Westernblot检测两株胰腺癌细胞PANC1和CFPAC1中Plk1表达。结果 胰腺癌组织中Plk1的阳性表达率为72、7％（24／33），其表达与胰腺癌患者的年龄、性别、临床分期、组织学分级、淋巴结转移、肿瘤的部位无显著相关（P〉0.05）。癌旁组织、慢性胰腺炎和良性胰腺肿瘤组织中Plk1的表达均为阴性。5例新鲜胰腺癌组织中都有Plk1mRNA的高表达，两株胰腺癌细胞中也有Plk1蛋白的表达。结论 Plk1在胰腺癌中的表达具有一定肿瘤特异性，有可能成为胰腺癌诊治的理想靶标。     

甲型H1N1与季节性H1N1流感病毒血凝素基因比较分析

目的分析泰安市2008～2009年度季节性流感与2009年度甲型H1N1流感病原学检测结果 ,比较季节性H1N1与甲型H1N1血凝素基因变异情况。方法选择国家级流感监测哨点医院以及暴发疫情的疫点,采集流感样病例的鼻咽拭子标本,通过RealtimePCR进行病毒检测,用MDCK细胞进行病毒分离,通过RT-PCR扩增血凝素HA1片段的基因并测序,利用生物信息学进行序列分析。结果 2008～2009年共检测鼻咽拭子标本283份,分离出流感病毒33株,分离阳性率为11.67%,其中季节性H1N1亚型31株。2009年5月1日～12月31日,检测鼻咽拭子标本996份,流感核酸检测阳性417份,阳性率为41.86%,其中甲型H1N1337份,季节性H1N1亚型1份。6株季节性H1N1病毒均在多个氨基酸位点上发生变异,与疫苗株A/Brisbane/59/2007(H1N1)比较,有11个位点发生了突变,其中5个位点位于抗原决定簇上;测序成功的6株甲型H1N1病毒在多个氨基酸位点发生变异,与疫苗株A/California/07/2009(H1N1)比较,有6个位点发生突变,其中1个位点位于抗原决定簇的B区。结论 2008～2009年度季节性H1N1为优势株,甲流暴发后,甲型H1N1成为绝对优势毒株。季节性H1N1分离株有多处氨基酸替换,抗原决定簇B区变异频繁;甲型H1N1病毒分离株的基因有变异,但关键位点第222位仍为D(天冬氨酸),与疫苗株相比抗原决定簇的关键位点变化不大。     

成人重症甲型H1N1流感8例临床分析

甲型H1N1流感是由甲型H1N1流感病毒引起的急性呼吸道传染病.人感染该病毒后会出现发热、咳嗽、头痛、疲劳及食欲不振等全身症状,但病死率较低,部分继发严重肺炎、急性呼吸窘迫综合征、肺出血、感染中毒症、呼吸衰竭及多器官损伤等,成为该病主要的死亡原因.我院急诊科2009年12月共收治甲型H1N1流感患者22例,其中重症8例(36.4%),现报道如下.     

Severe irinotecan-induced toxicity in a patient with UGT1A1*28 and UGT1A1*6 polymorphisms

Many studies have demonstrated the impact of UGT1A1 on toxicity of irinotecan. In particular, patients bear-ing UGT1A1*28 (TA 7/7) have a higher risk of severe neutropenia and diarrhea. Based on this, prescribers of irinotecan are advised that patients with UGT1A1*28 (TA 7/7) should start with a reduced dose of irinotecan, although a particular dose is not specified. Research in Asian countries has shown a lower incidence of UG-T1A1*28 (TA 7/7), while UGT1A1*6 (A/A) is more often found and is associated with severe irinotecan-related neutropenia. We report here a case of a metastatic colorectal cancer patient who is heterozygous for the UGT1A1*28 polymorphism (TA 6/7) as well as the UG-T1A1*6 polymorphism (G/A). The patient was treated with FOLFIRI for 9 cycles and underwent two irinote-can dose reductions according to pharmacokinetic data regarding exposure to the active metabolite, SN-38. Simultaneous heterozygous UGT1A1*28 and UGT1A1*6 polymorphisms may produce higher exposure to SN-38 and a higher risk of adverse effects related to irinote-can. Additional studies will be necessary to determine the optimal starting dose of irinotecan for patients with both UGT1A1*28 and UGT1A1*6 polymorphisms.     

Expression and significance of ICAM-1 and its counter receptors LFA-1 and Mac-1 in experimental acute pancreatitis of rats

AIM: To investigate the role of intercellular adhesion molecule-1 (ICAM-1) and its counter receptors LFA-1 and Mac-1 in acute pancreatitis (AP). METHODS: SD rats were allocated to AP group and control group randomly (25 rats each). AP was induced by infusion of 5% chenodeoxycholic acid into the pancreatic duct, followed by ligation of pancreatic duct. The rats were sacrificed at 1, 3, 6, 12 and 24 h after induction of pancreatitis. Five rats were sacrificed at one time point in the two groups before the blood and specimens from pancreas and lung were obtained. Serum amylase and ascitic fluid were measured at each time point. Expression of ICAM-1 at different time points was assessed by immunohistochemistry in pancreas and lung, and the expression of LAF-1 and Mac-1 on neutrophils at different time points was detected by flow cytometer. RESULTS: Induction of AP was confirmed by the serum levels of amylase and histological studies. The expression of ICAM-1 in pancreas increased significantly than that in the control group at all time points (P < 0.05 or P < 0.01), as well as the expression in lung except at 1 h. The expression of LFA-1 and Mac-1 on neutrophil in blood increased significantly in AP group than that in control group at several time points (P < 0.05 ox P < 0.01). The amount of ascitic fluid and serum amylase level of AP group increased significantly than that of control group at all time points (P < 0.05 or P < 0.01). Parallel to these results, a significant neutrophil infiltration was found in pancreas and lung tissues of AP group rats. CONCLUSION: Our findings suggest the important role for ICAM-1, LFA-1 and Mac-1 in mediating the development of AP from a local disease to a systemic illness. Upregulation of ICAM-1, LFA-1, Mac-1 and subsequent leukocyte infiltration appear to be significant events of pancreatic and pulmonary injuries in AP.     

SF-1和DAX-1在发育内分泌学的研究进展

核受体的作用极其广泛 ,类固醇生成因子 1(SF 1)和剂量敏感的性别转换综合征和先天性肾上腺发育不良症的关键基因 1(DAX 1)两种核受体家族是近年来发育内分泌学研究的热点之一 ,通过SF 1和DAX 1的动物模型及人类基因突变表现型阐述了在胚胎早期性发育和性分化中SF 1和DAX 1的作用可能通过相同途径相互拮抗 ,在垂体促性腺细胞和肾上腺能细胞中 ,两者在细胞及组织中的分布一致 ,通过下丘脑对垂体 性腺轴及垂体 肾上腺轴发育过程中不同时期的各个环节进行调控 ,本文对SF 1和DAX 1在内分泌器官发育过程中的分布、结构及其作用进行综述。     

RAF-1和ERK1在胃腺癌中的表达及意义

目的 检测胃腺癌组织中RAF-1、细胞外信号调节激酶1(ERK1)蛋白的表达特征,探讨二者的关系及临床意义.方法 选择80例胃腺癌术后留取的蜡块及临床资料作为观察组,选择60例正常胃黏膜组织的蜡块及临床资料作为对照组,应用免疫组织化学技术检测二组中RAF-1和ERK1蛋白的表达,探讨二者在肿瘤患者不同临床病理特征的表达差别及相关性.结果 观察组中RAF-1、ERK1蛋白表达的阳性率明显高于对照组,观察组中RAF-1、ERK1蛋白表达的阳性率均与肿瘤的转移、脉管浸润、临床分期及Ki67的表达密切相关.观察组中RAF-1和ERK1之间的表达具有正相关性.结论 胃腺癌组织中RAF-1和ERK1高表达,二者对病变的进展具有重要作用,二者可能具有内在协同作用,共同促进肿瘤浸润、侵袭和转移.     

肝脏表达的Niemann-Pick C1-Like 1调节小鼠的胆汁胆固醇浓度

目的:探讨人(NPC1L1)在小鼠肝脏的过表达对胆汁胆固醇重吸收的影响。方法:通过杂交NPC1L1基因敲除小鼠(L1-KO)与肝脏过表达人的NPC1L1的转基因小鼠(L1-Tg),获得无小鼠自身NPC1L1基因表达、在肝脏过表达或不表达人NPC1L1基因的小鼠(L1-Tgliv+和L1-Tgliv-)。给予L1-Tgliv+小鼠(实验组)及L1-Tgliv-小鼠(对照组)含0.2%胆固醇的饮食并测定2组小鼠胆汁的脂质分泌、血浆的脂质水平和粪便核心固醇的分泌。结果:与L1-Tgliv-小鼠相比,L1-Tgliv+小鼠的胆汁胆固醇水平降低了90%,同时粪便核心固醇的分泌也降低了10%,而血浆总胆固醇、游离胆固醇和胆固醇酯水平分别升高了38.6%、47.6%和36.6%。结论:小鼠肝脏NPC1L1的过表达可提高肝脏对胆汁中胆固醇的重吸收。     

HLA—DQB1、DPB1基因与1型糖尿病相关性研究

研究认为 ,1型糖尿病是遗传学上易感个体胰岛β细胞损害引起的自体免疫性疾病。基因标志是理论上预测 T1DM的最早指标。我们对 5 2例山东汉族 T1DM患者及 38例对照采用基于核酸序列的分型方法 (sequencing- based typing,SBT)进行HL A- DPB1基因分型 ,其中 32例患者及 2 3例对照测定了HL A- DQB1基因 ,探讨两者与 T1DM遗传易感性的相关性 ,结果报道如下。对象与方法1.研究对象 :T1DM患者 :5 2例 ,男 2 7,女 2 5 ,年龄 15～ 38岁 (2 5 .3± 6 .5岁 ) ,发病年龄 12～ 32岁 (2 2 .5± 8.9岁 )。正常对照 :健康查体者 38例 ,男 19,女…     

一例Rotor综合征SLCO1B1和SLCO1B3基因突变分析

目的初步探讨1例Rotor综合征患者的临床特点和SLCO1B1和SLCO1B3基因突变情况,从分子遗传学角度分析该疾病的发生机制。方法收集患者临床资料,从外周血白细胞提取基因组DNA,采用二代测序进行四千种已知单基因遗传性疾病筛查,用Sanger测序法分析验证二代测序发现的突变位点。结果患儿主要临床表现为反复皮肤及巩膜黄染,实验室检查示高胆红素血症,直接、间接胆红素双向增高。高通量测序发现患儿携带SLCO1B1基因c.1738 CT纯合突变和SLCO1B3基因c.360_481 del纯合突变。c.1738 CT突变为无义突变,已有文献报道,推测导致蛋白的第580位氨基酸密码子由精氨酸变为终止密码子。c.360_481 del突变为移码突变,蛋白编码区的第360至481位碱基缺失。该变异未见文献报道,也未见SNP数据库收录。此变异使蛋白缺失40个氨基酸的同时还造成开放阅读框移码,可能造成蛋白功能丧失。结论 SLCO1B1和SLCO1B3基因突变导致的有机阴离子转运多肽OATP1B1和OATP1B3功能缺陷是该Rotor综合征患者临床表现的分子遗传基础。     

老年重症/危重甲型H1N1流感40例临床分析

甲型H1N1流感作为急性呼吸道传染病,病原体为一种新型的甲型H1N1流感病毒.2009年,在墨西哥暴发,随后在全球迅速蔓延,数百万人感染,并有数千人死亡.感染甲型H1N1流感病毒的症状与普通季节性流感样症状类似,但病情严重者,恶化迅速,有肺炎,少数伴随呼吸衰竭、多脏器功能丧失,重者死亡.本文回顾性分析重症/危重症甲型H1N1患者主要临床特征,以研究早期症状与出现重症/危重症的相关性.