髓过氧化物酶和过氧化氢酶基因多态性及其酶活力与燃煤污染型地方性砷中毒关系的探讨

Objective To detect genetic polymorphism of myeloperoxidase (MPO) gene and catalase (CAT) gene and their activities, and to analyze their relationship with arsenic poisoning caused by coal-burning. Methods One hundred and thirty arsenic poisoning patients were chosen as case group in Jiaole Village, Xingren County, Guizhou Province(an endemic area). One hundred and forty healthy residents living in 13 km away were chosen as control group. Their blood was collected. Polymerase chain reaction-restriction fragment length polymorphism technique(PCR-RFLP) was used to detect polymorphism of MPO-463G/A and CAT-262C/T. Ultraviolet spectmphotometer method was used to detect myeloperoxidase activity. Chromatometry method was used to detect catalase activity. Results The genotype frequency of MPO-463G/A at GG, GA, AA site was 47.24%(60/127), 44.09%(56/127),8.67% (11/127) in case group and 42.34% (58/137),48.17% (66/137)1,9.49% (13/137) in control group, respectively. The difference between the two groups was not significant(χ2 = 0.642, P > 0.05). The genotype frequency of CAT-262C/T, at CC, CT, TT site was 65.60%(82/125),28.80%(36/125),5.60%(7/125) in case group and 76.51%(101/132), 18.94% (25/132) ,4.55% (6/132) in control group, respectively, without significant difference (χ2 =3.845, P>0.05). The relationship between polymorphism of MPO-463G/A and CAT-262C/T and the risk of arsenic poisoning was not found in this study(ORadj= 1.36, 95%CI: 0.74-2.50 for MPO; ORadj=1.35, 95%CI: 0.69-2.63 for CAT). The activities of MPO and CAT were (25.30±8.70)U/L and (2.80± 1.09)×103 U/L in case group, while (22.76±7.59)U/L and (3.90±1.01)×103U/L in control group with a significant difference(F=0.760 for MPO, F=0.855 for CAT, all P < 0.05). The genotype of MPO-463G/A and CAT-262C/T was not found to have relationship with the activities of MPO, CAT(F=1.312,2.822 for MPO; F= 0.151,0.036 for CAT, P>0.05). Conclusions Genetic polymorphism of MPO-463G/A and CAT-262C/T is not found to have relationship with arsenic poisoning. Arsenic can lead to the change of MPO and CAT activity, which, however, may not be affected by MPO-463G/A and CAT-262C/T polymorphism.     

燃煤型砷中毒患者p16基因缺失及启动区CpG岛甲基化的观察

目的检测燃煤型砷中毒(地砷病)患者p16基因缺失及启动区甲基化的变异,探讨p16基因异常改变在地砷病发生发展乃至癌变过程中的作用。方法采用多重PCR法对103例砷中毒患者和110例正常对照人群p16基因第1、第2外显子缺失情况进行检测;同时采用甲基化特异性PCR法(MSP)和PCR产物测序技术对95例砷中毒患者和100例正常对照人群p16基因启动区甲基化情况进行分析。结果(1)正常对照组p16基因缺失率为5．45％;病例组为14．56％,以第2外显子缺失为主,其中轻、中、重度中毒组p16基因缺失率分别为9．09％、12．50％和19．51％;非癌变组和癌变组p16基因缺失率分别为8．70％和38．89％。以上差异有统计学意义(P<0．05或P<0．01)。(2)对照组p16基因甲基化阳性率为2．00％;病例组为42．11％,其中轻、中、重度中毒组p16基因甲基化阳性率分别为26．32％、41．67％和50．00％;非癌变组和癌变组阳性率分别为21．74％和55．56％。以上差异均有统计学意义(P<0．05或P<0．01)。结论p16基因缺失及启动区甲基化在地砷病的发生发展乃至癌变过程中起重要作用。     

军队医院创建地方等级医院的实践与体会

安顺73医院始建于1963年,原为部队驻军医院,现隶属贵州省军区陆军预备役师。为加快医院的全面建设和发展,进一步提升医院的综合实力和发展空间,经报请军地上级主管部门批准,医院申请参加了当地二级甲等医院（以下简称“二甲医院”）的评审。从2010年8月开始,医院精心准备、周密安排、认真实施,经过组织动员、对照检查、整改完善、巡查改进、评审验收5个阶段,圆满完成创建任务,2011年11月顺利通过当地卫生部门组织的二甲医院评审。笔者作为创建工作的组织者及特聘专家,现将创建、评审的主要做法和体会简要报告如下。     

髓过氧化物酶和过氧化氢酶基因多态性及其酶活力与燃煤污染型地方性砷中毒关系的探讨

Objective To detect genetic polymorphism of myeloperoxidase (MPO) gene and catalase (CAT) gene and their activities, and to analyze their relationship with arsenic poisoning caused by coal-burning. Methods One hundred and thirty arsenic poisoning patients were chosen as case group in Jiaole Village, Xingren County, Guizhou Province(an endemic area). One hundred and forty healthy residents living in 13 km away were chosen as control group. Their blood was collected. Polymerase chain reaction-restriction fragment length polymorphism technique(PCR-RFLP) was used to detect polymorphism of MPO-463G/A and CAT-262C/T. Ultraviolet spectmphotometer method was used to detect myeloperoxidase activity. Chromatometry method was used to detect catalase activity. Results The genotype frequency of MPO-463G/A at GG, GA, AA site was 47.24%(60/127), 44.09%(56/127),8.67% (11/127) in case group and 42.34% (58/137),48.17% (66/137)1,9.49% (13/137) in control group, respectively. The difference between the two groups was not significant(χ2 = 0.642, P > 0.05). The genotype frequency of CAT-262C/T, at CC, CT, TT site was 65.60%(82/125),28.80%(36/125),5.60%(7/125) in case group and 76.51%(101/132), 18.94% (25/132) ,4.55% (6/132) in control group, respectively, without significant difference (χ2 =3.845, P>0.05). The relationship between polymorphism of MPO-463G/A and CAT-262C/T and the risk of arsenic poisoning was not found in this study(ORadj= 1.36, 95%CI: 0.74-2.50 for MPO; ORadj=1.35, 95%CI: 0.69-2.63 for CAT). The activities of MPO and CAT were (25.30±8.70)U/L and (2.80± 1.09)×103 U/L in case group, while (22.76±7.59)U/L and (3.90±1.01)×103U/L in control group with a significant difference(F=0.760 for MPO, F=0.855 for CAT, all P < 0.05). The genotype of MPO-463G/A and CAT-262C/T was not found to have relationship with the activities of MPO, CAT(F=1.312,2.822 for MPO; F= 0.151,0.036 for CAT, P>0.05). Conclusions Genetic polymorphism of MPO-463G/A and CAT-262C/T is not found to have relationship with arsenic poisoning. Arsenic can lead to the change of MPO and CAT activity, which, however, may not be affected by MPO-463G/A and CAT-262C/T polymorphism.     

基于UHPLC-LTQ-Orbitrap MS代谢组学的不同产地酸枣仁化学成分差异性比较

目的 探究不同产地酸枣仁化学成分的差异及分布规律。方法 基于代谢物信息公共数据库,利用超高效液相色谱-线性离子阱/静电场轨道阱组合式高分辨质谱法（UHPLC-LTQ-Orbitrap MS）结合代谢组学多元统计技术,对检测的一级谱、二级谱数据进行定性分析,并采用主成分分析（PCA）、正交偏最小二乘法-判别分析（OPLS-DA）对化合物进行分析,确定不同产地酸枣仁的差异代谢物。结果 通过对照品指认、文献比对及高分辨质谱数据解析,共鉴定酸枣仁中49个化合物和9个新成分。其中,山柰酚-3-O-芸香糖苷、6?-对羟基苯甲酰斯皮诺素及美洲茶酸在不同产地间差异有统计学意义。结论 通过UHPLC-LTQ-Orbitrap MS,发现不同产地酸仁枣化学成分的整体差异性较大,该结果可为进一步研究掺假酸枣仁及不同产地酸枣仁的质量评价提供参考。     

基准剂量在燃煤砷暴露人群肝损害研究中的应用及其意义探讨

目的：探讨燃煤砷暴露人群肝损害的生物接触限值,筛选监测砷致肝损害的敏感生物学标志。方法：以燃煤污染型地方性砷中毒病区118例砷暴露者为砷暴露组,非砷污染区50例居民为对照组。以尿砷、发砷（Ag—DDC法测定）作为接触指标;总胆汁酸（TBA,酶循环法测定）、谷胱甘肽硫转移酶（GSTs,     

基准剂量在燃煤砷暴露人群肝损害研究中的应用及其意义探讨

目的 探讨燃煤砷暴露人群肝损害的生物接触限值,筛选监测砷致肝损害的敏感生物学标志.方法 以燃煤污染型地方性砷中毒病区118例砷暴露者为砷暴露组,非砷污染区50例居民为对照组.以尿砷、发砷(Ag-DDC法测定)作为接触指标;总胆汁酸(TBA,酶循环法测定)、谷胱甘肽硫转移酶(GSTs,化学比色法测定)和γ-谷氨酰转肽酶(γ-GT,重氮试剂比色法测定)作为肝细胞损害效应指标;血清透明质酸(HA)、Ⅲ型前胶原(PC-Ⅲ)和Ⅳ型胶原(Ⅳ·C)作为肝纤维化效应指标(均用放射免疫法测定).运用基准剂量(BMD)法计算各效应指标对应的尿砷和发砷的BMD及其95%可信区间下限值(BMDL),并根据BMD和BMDL大小判断各效应指标的敏感性.结果 砷暴露组的砷接触指标尿砷(98.50 mg/kg Cr)、发砷(7.42 ms/kg)和肝损害效应指标TBA、GSTs、γ-GT、HA、PC-Ⅲ、Ⅳ·C的几何均数(6.78 μmol/L、22.05 U/L、48.04 U/L、85.19μg/L、89.76μg/L、86.85μg/L)均明显高于对照组的尿砷(22.98 mg/kg Cr)、发砷(1.28 ms/kg)和肝损害效应指标(4.63μmol/L、13.76 U/L、36.45 U/L、54.62μg/L、74.45μg/L、54.81μg/L,P<0.01),尿砷、发砷与肝损害效应指标间存在剂量-效应关系(P<0.05或<0.01).尿砷的BMD和BMDL范围分别为49.53～101.96、39.02～70.15mg/kg Cr,发砷的BMD和BMDL范围分别为3.04～5.02、2.36～3.25 mg/kg.肝细胞损害指标的敏感顺序均为GsTs>TBA>γ-GT;而肝纤维化指标的敏感顺序均为HA>Ⅳ·C>PC-Ⅲ.结论 根据肝损害生物学标志的最低尿砷、发砷BMDL值,结合当地健康人群尿砷、发砷平均值,建议燃煤砷暴露者肝损害的生物接触限值尿砷为35.00 mg/kg Cr,发砷为2.50 ms/kg.GSTs、TBA、γ-GT和HA、Ⅳ·C、PC-Ⅲ分别能不同程度地反映燃煤砷污染对人体的肝细胞损害和肝纤维化情况:GSTs、HA分别是其相对最敏感的生物学标志.     

碳酸锂对抗 AFB1 诱导大鼠肝癌过程中 MDA、 4-HNE 的表达及意义

背景与目的:采用免疫组化法检测碳酸锂(Li2CO3)对抗黄曲霉素B1(AflatoxinB1,AFB1)诱导大鼠肝癌过程中丙二醛(Malondialdehyde,MDA)、4_羟壬烯醛(4_hydroxynonenal,4_HNE)的表达情况,探讨Li2CO3 的抗癌作用及其机制。材料与方法:144只Wistar大鼠随机分为A、B、C、D4组,分别为阴性、阳性对照及Li2CO3 同时给药和Li2CO3 先期给药组。于实验第6、9、10周分批断头处死动物,取动物的肝脏进行肝组织形态学检查及免疫组织化学染色。结果:C、D两组动物健康状况明显改善,肝癌前病变程度明显减轻;MDA、4_HNE在诱癌早期(实验第6周)即有表达,第10周显著增高,并呈B组>C组>D组>A组的趋势。结论:Li2CO3 具有明显地对抗和抑制化学诱导肝癌过程中的脂质过氧化(Lipidperoxidation,LPO)作用;免疫组化法检测MDA、4_HNE可敏感而特异的反映机体氧化损伤程度,有助于肝癌的早期发现及病变进展的动态观察。     

燃煤型地方性砷中毒患者外周血淋巴细胞亚群的改变及意义

[目的]了解燃煤污染型地方性砷中毒患者外周血淋巴细胞亚群的变化,探讨其在砷中毒发生发展中的意义。[方法]中毒组为53名燃煤型砷中毒患者,暴露组为7名砷暴露者,对照组为32名健康者。从中毒组选取22例患者（一般病变组11例,癌前病变和癌变组11例）,采用流式细胞分析技术检测其外周血T淋巴细胞（CD3^＋）、辅助性T淋巴细胞（Th细胞,CD3^＋CD4^＋）、抑制性T淋巴细胞（Ts细胞,CD3^＋CD8^＋）、Th／Ts比值（CD3^＋CD4^＋／CD3^＋CD8^＋）、B淋巴细胞（CD3^-CD19^＋）和自然杀伤细胞[NK细胞,CD3^-CD（16＋56）^＋]的改变情况,并比较不同临床分度砷中毒患者和不同皮肤病损患者上述指标的差异。[结果]与对照组比较,轻、中、重度中毒组患者外周血CD3^＋、CD3^＋CD4^＋和CD3^-CD19^＋淋巴细胞百分比均降低（P〈0.01或P〈0.05）,暴露组CD3^＋CD4^＋和CD3^-CD19^＋淋巴细胞百分比降低（P〈0.05）;重度中毒组CD3^-CD（16＋56）^＋细胞百分比高于对照组和暴露组（P〈0.05）;与一般病变组比较,癌前病变和癌变组外周血CD3^＋、CD3^＋CD4^＋和CD3^-CD19^＋淋巴细胞百分比均降低（P〈0.05）;[结论]Th细胞和B淋巴细胞可能是燃煤型砷污染致人体免疫损伤的早期靶点。燃煤型砷中毒患者外周血中Th细胞和B淋巴细胞百分比的降低可能在砷中毒的发生发展过程中起一定作用。