贵州燃煤污染型地氟病病区居民血糖水平改变

燃煤污染型地方性氟中毒(简称地氟病)是中国特有的地氟病类型,由于当地燃煤中有较高氟含量,通过烘烤粮食、取暖等方式污染食物和空气,导致人体发生慢性氟中毒[1].贵州是人群最多和病情最重的燃煤污染型地氟病病区[1-2],近年来,各级相关部门对贵州省燃煤污染型地氟病病区投入了大量的人力和物力,通过改良炉灶、健康教育和改善卫生条件等综合措施来防治地氟病,取得较为显著的效果[3],极大地减少了新病人的产生,并对地氟病患者病情有所缓解.     

载脂蛋白J基因多态性与2型糖尿病相关性分析

目的探讨载脂蛋白J(ApoJ)外显子3、4、7、8基因多态性与2型糖尿病(T2DM)的关系。方法2002-11~2003-06对贵阳医学院附属医院,利用聚合酶链式反应-变性梯度凝胶电泳技术(PCR-DGGE)对61例T2DM患者、60例正常对照者的ApoJ外显子3、4、7、8基因进行基因筛查,并对异常条带进行测序分析。结果我国ApoJ基因外显子7缺失与插入(DVSI)基因型在T2DM与正常对照组间分布差异显著(P<0.01),ApoJ外显子3、4、8差异无显著性(P>0.05)。结论我国2型糖尿病患者ApoJ外显子7基因插入与缺失多态性可能与2型糖尿病发病有关联,是2型糖尿病的危险因素之一。     

贵州省从江县侗族人群葡萄糖-6-磷酸脱氢酶基因突变型的检测

目的了解贵州省从江县侗族葡萄糖-6-磷酸脱氢酶(Glucose-6-phosphate dehydrogenase,G6PD) 缺乏症的发生率、基因突变类型及特点。方法对贵州省从江县侗族524人采用四氮唑蓝定性法进行G6PD缺乏症初筛、G6PD／6PGD比值法验证．再经自然引物及错配引物介导的聚合酶链反应／限制性酶切分析法检测中国人常见的9种基因突变型,对于未定型采用变性梯度凝胶电泳法(DGGE)检查外显子2、8、9、12基因突变情况。结果 G6PD缺乏症34例,检出率为6．49％,其中检出G1388A突变4例、C592T突变18例。未定型12例经DGGE检测外显子突变情况,未发现突变,有待于进一步对其余外显子进行研究。结论贵州省从江侗族是G6PD缺乏症的高发区。592 C→T突变型为该地该民族常见突变型,而不是中国人常见的G1376T、G1388A或A95G突变型。此次基因突变型调查为了解贵州省少数民族G6PD缺乏症的分布特征提供了原始数据。     

贵州江口土家族男性葡萄糖-6-磷酸脱氢酶缺陷症的基因频率调查

目的调查贵州省江口土家族男性葡萄糖-6-磷酸脱氢酶(G6PD)缺陷症的基因频率。方法2002年10—11月随机选取227名江口当地土家族男性居民,采用四氮唑蓝(NBT)纸片定性法初筛G6PD缺陷症,G6PD/6PGD比值法定量确诊。结果对贵州省江口县227名土家族男性进行筛查,共检出17例G6PD缺陷阳性,其基因频率为0·0749。结论通过对贵州土家族G6PD缺乏症基因频率的调查和分析,初步了解了贵州省土家族G6PD缺乏症的分布特征。为该地区预防该疾病、指导临床、提高少数民族的素质及研究该民族起源提供了一定的依据。     

贵州省地方性氟中毒地区人群GSTP1基因Ile105Val位点多态性分析

Objective To investigate plasma glutathione S-transferase(GSTs) activity and GSTP1 gene Ile105Val polymorphism in Bijie City, Guizhou Province, a coal-burning fluorosis endemic area. Methods One hundred and sixty villagers from Yachi Twon using non-improved cooking stoves were selected as the non-intervened group in Bijie City, Guizhou Province where coal-burning fluorosis was prevailing; 153 villagers as the intervented group were chosen from Changchun Twon, where cooking stoves were improved; 151 villagers were served as the control group from Baiyunshan Twon, Changshun County without endemic fluorosis. The activity of GSTs was tested by colorimetric analysis with spectrophotometer. The genotype of the GSTP1 gene Ile105Val polymorphism, presenting as either homozygous wild-type (AA), or heterozygous mutation type (AG), or homozygous mutation type (GG), was detected through the PCR-RFLP procedure. Results The activity of GSTs in plasma of non-intervened group [(12.44±4.97) kU/L]was significantly lower than that of intervened group (P < 0.05), and that of intervened group[(20.78±6.20)kU/L]was significantly lower than that of control group[(24.30±6.27)kU/L, P< 0.05]. The difference of the enzyme activity of three groups were statistically significant (F = 51.71, P < 0.05), but this enzyme activity did not vary significantly in each sex of each grnup(P > 0.05). Compared intervened group [AA:67.3%(103/153), AG:29.4%(45/153),GG:3.3%(5/153)]and non-intervened group[AA:66.9%(107/160), AG:30%(48/160), GG:3.1%(5/160)]with control group[AA:74.8%(113/151), AG:25.2%(38/151), GG:0 (0/151)], the Ile105Val polymorphism site of GSTP1 gene had significant difference(χ2= 6.04,6.07, both P< 0.05), but not significant between intervened and non-intervened groups(χ2 = 0.02, P>0.05). Conclusions Fluorosis can decrease the activity of GSTs and introduce the GSTP1 gene Ile105Val polymorphism, intervention with the fluorine intake will improve the effect of fluoride on the body.     

慢性氟中毒大鼠脑组织中c-Jun氨基末端激酶表达水平观察

目的 观察慢性氟中毒大鼠脑组织中c-Jun氨基末端激酶(JNK)信号转导激酶表达变化,进一步揭示慢性氟中毒神经损伤的分子机制.方法 SD大鼠随机分为3组:对照组、低氟组、高氟组,每组24只,饮用水含氟量分别为＜0.5和5.0、50.0 mg/L,实验期为6个月.用氟离子选择电极法测定大鼠尿氟及血氟,用Western blotting和免疫组织化学方法检测脑组织中JNK信号转导激酶的表达和分布,并分析血氟与活化的JNK激酶的相关关系.结果低氟组和高氟组大鼠尿氟[(2.56±0.91)、(5.73±3.14)mg/L]和血氟[(0.36±0.14)、(0.50±0.18)mg/L]均较对照组[(0.92±0.30)、(0.12±0.07)mg/L]升高(P均＜0.05).高氟组(1.74±0.69)脑组织phospho-JNK表达高于对照组(1.00±0.37)和低氟组(1.20±0.28,P均＜0.05);total-JNK蛋白表达水平3组间比较,差异无统计学意义(F=0.046,P＞0.05).phospho-JNK、total-JNK阳性表达神经元主要集中在皮质、海马和背侧丘脑,其中高氟组大鼠phospho-JNK在顶叶皮质(119.3±14.1)、枕叶皮质(112.7±5.4)、海马CA3区(100.6±8.9)、背侧丘脑(117.8±10.4)及橄榄核(112.6±5.9)中阳性表达较对照组(104.1±8.9、106.6±9.6、106.6±9.7、108.9±6.4、100.3±8.4)和低氟组(96.7±17.1、102.5±8.3、106.4±6.5、110.2±9.3、102.4±4.7、102.5±9.8)明显增高(P均＜0.05),而total-JNK在各组大鼠脑组织中阳性表达分布未见明显改变(P均＞0.05).相关分析结果发现,随大鼠血氟升高,脑组织中phospho-JNK表达呈增高趋势,二者存在正相关关系(r=0.677).结论慢性氟中毒导致脑组织中磷酸化JNK表达改变,并与机体中氟蓄积量存在相关关系,这些改变可能与慢性氟中毒导致的神经损伤有关系.     

燃煤污染型氟中毒大鼠学习记忆能力及胆碱酯酶活性改变

Objective To observe the influence of coal burning fluorosis on learning and memory ability in rats and reveal its possible mechanisms. Methods Healthy 48 SD rats were divided into control, low-fluoride and high-fluoride group. All rats in fluoride exposed groups were fed with the eom polluted by drying processes with burning coal containing high level of fluoride obtained from the endemic fluorosis area to produce the animal model of fluorosis. The experiment period were 3,6 mouths, respectively. The ability of leaning and memory was measured by Morris test and cholinesterase activity detected by photometric method at 3 or 6 month after experiment, respectively. Results Fluoride contents signifieantlly influenced the escape latency, the numbers of crossing the platforms and the time of staying the platforms(the value of F was 29.29,6.47,6.50, respectively, P<0.01).In addition, the numbers of crossing the platforms and the time of staying the platforms were influenced by the exposed time(the value of F was 16.11,45.59, P<0.01). Furthermore, the fluoride contents and the exposed time had an interaction between the numbers of crossing the platforms and the time of staying the platforms (the value of F was 4.67,5.68, P<0.05 or<0.01). Three months after the experiment, the mean values of escape latency [(14.71± 4.85)s] of rats in highly fluoride exposed group were significantly prolonged as compared with controls [(9.28±4.22)s]; 6 month after the experiment, the mean values of escape latency[(12.42±8.03)s, (17.48± 8.05)s] of rats in both groups exposed to fluoride were significantly prolonged as compared to controls [(7.04± 3.29)s, P<0.05]. The decreased numbers of crossing the platforms[(1.62±0.87)number] and the declined time of staying the platforms[(16.70±5.02)s] were found in the rats exposed to high fluoride as compared to controls [(3.53±1.67 )number, (23.33±5.35)s, P<0.05]. The fluoride contents obviously influenced the activities of acetylcholinesterase and butylcolinesterase (the value of F was 12.83,13.27, P<0.01). On the other hand, the times of breeding also influnced the activities of butylcolinesterase (the value of F was 16.26, P<0.01). In 3 months of the experiment, the activities of butylcolinesterase [(0.55±0.12)kU/g] in low fluoride exposed group were significantly decreased in comparison with controls[(0.73±0.10)kU/g, P<0.05]. The activities of acetylcholinesterase[(0.62±0.42)kU/g] and butylcolioesterase[(0.58±0.10)kU/g] in high fluoride group were significantly decreased as compared to eontrois[(1.41±0.52), (0.73±0.10)kU/g, P<0.05]. The correlation analysis showed that there was a negative correlation between the cholinesterase and the escape latency(r=-0.68, P< 0.01), and a positive correlation between the cholinesterase and the time of staying the platforms(r=0.57, P< 0.01). Conclusions The ability of learning and memory in rats with coal buring fluorosis was decreased, which might be connected to the decreased activity of cholinesterase in a dose-effect correlation.     

贵州省兴仁县燃煤型砷中毒患者金属硫蛋白-1B和-1X mRNA表达

目的 观察金属硫蛋白(MT)-1B、MT-1X mRNA在燃煤型砷中毒患者外周血细胞中的表达.方法 采集贵州省兴仁县燃煤型砷中毒患者(砷中毒组)及非病区正常人(对照组)外周血,各50例,提取及纯化总RNA,采用实时荧光定量PCR(real-time PCR)技术测定血细胞中MT-1B、MT-1X mRNA的表达,以β-actin作为质控.结果 对照组外周血细胞MT-1B、MT-1X mRNA表达为0.15±0.25、44.39±27.67,砷中毒组为0.03±0.10、7.36±7.34,组间比较差异有统计学意义(t值分别为3.15、9.15,P＜0.05).结论 砷中毒降低MT-1B、MT-1X在外周血细胞中的表达.     

贵州省荔波瑶族、布依族、汉族β-地中海贫血血液学筛查分析

β-地中海贫血是世界上最常见和发病率最高的一种单基因遗传性溶血性疾病，迄今已发现100多种突变类型，因而β-地中海贫血病的研究一直受到国内外的广泛关注和重视。据世界卫牛组织报告，约有一亿以上的人口携带该病的突变基因。其分子病理有极大的异质性，基因类型及频率分布具有极大的地域和民族差异。在我国广东、广西、云南、贵州等省β-地中海贫血发病率在2％左右，其中贵州少数民族地区的发病率高达4．72%。故在少数民族人群中开展β-地中海贫血研究，不仅可大大充实不同民族遗传多样性研究和疾病基因分析研究的内容，丰富人类基因组多样性研究的内涵，而且对该地人群进行婚育指导、产前诊断、提高人口素质亦有重要意义。因此笔者对贵州省荔波县茂兰镇瑶族、布依族、汉族进行了β-地中海贫血群体调查，结果如下。