Effects of α3 neuronal nicotinic acetylcholine receptor on cell apoptosis and p38 MAPK signal transduction pathway in SH-SY5Y cells / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the effects of α3 neuronal nicotinic acetylcholine receptor (nAChR) on apoptosis and p38 signal transduction pathway in SH-SY5Y cells and to assess the roles of α3 nAChR in the pathogenesis of Alzheimer's disease (AD).</p><p><b>METHODS</b>The levels of α3 nAChR mRNA and protein were measured by real-time PCR and Western blot, respectively, in SH-SY5Y cells transfected with α3 nAChR siRNA. The mRNA level of bcl-2 and bax was measured by the real-time PCR. The siRNA transfected SH-SY5Y cells and control were then treated with 10 µmol/L Aβ25-35 for another 48 h, and the change in apoptotic rate and the levels of p-p38 and p38 were measured by flow cytometry and Western blot. Subsequently these SH-SY5Y cells were exposed to a blocker of p38 protein, and the apoptotic rate was measured again.</p><p><b>RESULTS</b>Compared to the controls, the expression of α3 nAChR at mRNA and protein levels in the SH-SY5Y cells transfected with α3 nAChR siRNA decreased by 95% and 86%, respectively; the mRNA levels of bax increased 2.11 times and that for bcl-2 decreased 0.53 times. The apoptotic rate was unaffected (3.40% ± 0.20%); but it increased after Aβ25-35 treatment (24.52% ± 1.59%); the level of p-p38 protein also increased by 178% in the α3 nAChR inhibited cells treated with Aβ25-35. Compared to controls, the Aβ25-35-treated SH-SY5Y cells and the Aβ25-35-treated and siRNA-transfected cells both showed a reduction in apoptosis after treatment with p38 blocker, especially in the former.</p><p><b>CONCLUSION</b>The siRNA silencing of α3 nAChR mRNA may enhance the effect of Aβ25-35 on the cell apoptosis by increasing the levels of p38 protein and bax mRNA and decreasing the level of bcl-2 mRNA, which may play a role in the pathogenesis of AD.</p>

A study of interleukin-10 gene polymorphisms in Miao, Dong and Buyi ethnics of Guizhou / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate allelic frequencies of interluekin-10 (IL-10) gene promoter in Miao, Dong and Buyi ethnics of Guizhou.</p><p><b>METHODS</b>TaqMan MGB-based real-time PCR was used to determine the genotypes of IL-10 -819 and IL-10 -592 in 589 Miao, Dong and Buyi ethnics of Guizhou.</p><p><b>RESULTS</b>The allelic frequency of IL-10 -819 in Miao ethnics was significantly different from those in Dong or Buyi ethnics. Allelic frequencies of IL-10 -592 in Miao ethnics was significantly different from those in Dong or Buyi ethnics. In Miao, Dong and Buyi ethnics, the distributions of genotype frequencies of IL-10 -819 and IL-10 -592 were statistically different from Han ethnics from Guizhou and Taiwan of China as well as South Koreans.</p><p><b>CONCLUSION</b>There is a heterogeneity in the frequencies of polymorphisms of IL-10 promoter among different ethnic groups.</p>

Association of vitamin D receptor gene polymorphisms with susceptibility to coal-burning borne fluorosis in Guizhou province / 中国地方病学杂志

ObjectiveTo observe the distribution of vitamin D receptor(VDR) gene polymorphisms in coal-burning borne fluorosis in Guizhou province and investigate the relationship between VDR gene polymorphisms and the susceptibility to coal-burning borne fluorosis.MethodsOne hundred and fifty villagers from non-improving cooking stove villages were selected as a non-intervention group in Bijie area,Guizhou province where coal-burning borne fluorosis was prevailing; 150 villagers were chosen from cooking stove improved villages as a intervention group; 150 villagers were selected from non-endemic area Changshun county as a control group.DNA was extracted from peripheral blood samples of these people.Genotype of VDR gene Bsm Ⅰ and Fok Ⅰ loci were detected using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP).ResultsDistribution of Bsm Ⅰ polymorphism site of VDR gene of control group [AA:19.3％ (29/150),AG:39.3％ (59/150),GG:41.3％(62/150)],was compared with that[AA:4.7％(7/150),AG:14.0％(21/150),GG:81.3％(122/150)] of the non-intervention group and that[AA:7.3％(11/150),AG:23.3％(35/150),GG:69.3％(104/150)] of intervention group,and the difference was statistically significant(X2 =56.6,P ＜ 0.05).The frequency of VDR-Fok Ⅰ loci in non-intervention group [TT:29.3％(44/150),TC:55.3％(83/150),CC:15.3％(23/150)] and intervention group [TT:32.7％(49/150),TC:55.3％(83/150),CC:12.0％(18/150)] was compared with that [TT:45.3％(68/150),TC:48.7％(73/150),CC:6.0％(9/150)] of control group,and the difference was statistically significant(X2 =11.9,P ＜ 0.05).Univariate analysis showed that individuals carrying the GG genotype had increased risk of suffering fluorosis than individuals carrying the AA and AG genotypes(OR values were 6.2,3.2,all P ＜ 0.05),while carrying the TC and CC genotype had increased risk of suffering fluorosis than individuals carrying the TT genotype (OR values were 1.3,2.8,1.3,2.1,all P ＜ 0.05).ConclusionVDR gene polymorphisms may be one of the predisposing factors of coal-burning borne fluorosis.

Genetic association between interleukin-10 promoter microsatellite polymorphisms and hepatitis B virus infection in Yi, Yao and Han ethnic populations of Guizhou province / 中华流行病学杂志

Objective To investigate the association between interleukin-10 (IL-10) gene promoter microsatellite polymorphisms and the susceptibility to hepatitis B virus infection in Han,Yi and Yao ethnicities in GuiZhou province.Methods 500 volunteers were selected from Guizhou province.Ailelic frequency of IL-10.G and IL-10.R loci was identified by short tandom repeat polymerase chain reaction.The relativity between allelic frequency and HBV infection was analyzed.Results Genotype data from H-W analysis on all the IL-10 polymorphisms indicated that it was a random distribution.Very high HBV infection rates were found in the native ethnic minorities of Guizhou province.The overall HBV infection rate among the total population was 67.00％,with the HBV infection rates of Yi nationality in Weining,Yi nationality in Qianxi,Yao nationality in Libo and Han nationality in Libo as 51.85％,42.86％,79.52％ and 84.30％,respe~vely.The polymorphisms distribution of IL- 10.G and IL- 10.R were statistically different among the ethnic groups (P＜ 0.05 ).The polymorphisms distribution of IL-10.R had no significant difference between HBV infection group and non-infection group,as well as among HBV natural removal group and non-infected group in all the ethnic groups.The frequency of IL-10.G 459 bp (19CA) was significantly higher in non-infection group than in the infected group (P＜ 0.05 ).The frequency of IL-10.G 471 bp (25CA) was significantly higher in the non-infection group than in the HBV natural removal group(P＜0.05).The polymorphisms distribution of IL-10.G did not show significant difference between the HBV infection group and the HBV natural removal group in all the ethnic groups.We did not find any differences in allelic and genotypic frequencies of IL-10.G between infection group and non-infection group in Yi nationality in Weining,and Yao nationality in Libo (P＞0.05),as well as HBV natural removal group and non-infected group (P＞0.05).Conclusion The polymorphisms distribution of IL-10.R and IL-10.G did not show significant difference in Yi,Yao and Han ethnics population living in Guizhou province.IL-10.G seemed to influence the susceptibility of HBV infection in Han,Yao and Yi ethnics population of Guizhou province.

Relationship between-262C/T and-21A/T polymorphism of catalase gene and coal-burning borne fluorosis / 中国地方病学杂志

Objective To explore the relationship between -262C/T and -21A/T polymorphisms of catalase(CAT) gene and coal-burning borne fluorosis. Methods In 2007, 150 villagers were taken as a nonintervention group in Bijie city from the village of coal-burning borne fluorosis areas with unchanged cooking stoves;150 villagers were taken as the intervention group from the town of Changchun county where cooking stoves changed; 150 villagers were taken as control from non-endemic fluorosis areas in Baiyun town of Changshun county.PCR-restriction fragment length polymorphism were employed to detect genotypes of CAT-262C/T and CAT-21A/T polymorphism of CAT gene. Results The genotypic frequencies of CAT-262C/T and CAT-21A/T in nonintervention group,intervention group and control group were in line with Hardy-Weinberg equilibrium law (P＞ 0.05 ).The genotypes of CC and CT were detected while no TT were detected for CAT-262C/T polymorphism; the genotypes of AA, AT and TT were detected for CAT-21A/T. The genotype frequencies of CAT-262 CC, CT in control group, intervention group and non-intervention group were (89.33%(134/150), 10.67%(16/150); 88.67%(133/150), 11.33% (17/150),93.33% (140/150),6.67% (10/150), respectively. The gene frequency of C in control group, intervention group and non-intervention group were (94.67% (284/300), 94.33% (283/300),96.67%(290/300), respectively. The gene frequency of T in control group, intervention group and non-intervention group were 5.33%(16/300), 5.67%(17/300), 3.33%(10/300), respectively. The genotype frequencies of CAT-21 AA,AT and TT in control group, intervention group and non-intervention group were 48.67%(73/150),46.00%(69/150),5.33%(8/150) ,52.67%(79/150) ,38.00%(57/150) ,9.33% (14/150) ,51.33%(77/150) ,38.00%(57/150), 10.67%(16/150), respectively. The gene frequency of A in control group, intervention group and non-intervention group were 71.67%(215/300),71.67%(215/300),70.33%(211/300), respectively. The gene frequency of T in control group, intervention group and non-intervention group were 28.33% (85/300),28.33% (85/300),29.67% (89/300),respectively. CAT-262C/T and CAT-21A/T genotype and allele frequencies in the control group, the intervention group and non-intervention group showed no significant differences in the distribution(x2= 0.331,0.336, all P ＞0.05 ). Conclusion CAT-262C/T and CAT-21A/T polymorphism is not associated with coal-burning borne fluorosis.

Expression of glutathione S-transferase mRNA in peripheral blood of the population in coal-burning fluorosis area / 中国地方病学杂志

Objective To investigate expression of glutathione S-transferase(GST) mRNA in peripheral blood of the population in coal-burning fluorosis area and to evaluate the effect of comprehensive control intervention. Methods Fifty samples of peripheral blood from patients in the coal-buring fluorosis area in Bijie county of Guizhou province were selected as fluorasis group and 50 samples of peripheral blood from patients in area with comprehensive management were selected as intervention group, respectively. Fifty samples from non-endemic fluorosis area were selected as the control group. Total RNA from blood was extracted and purified by the Trizol- Phenol-Chloroform one-step method. Expression of GST mRNA was detected by using SYBR Green I real-time fluorescence quantitative PCR. Results The data of GST mRNA in fluorosis group, intervention group and control group was 38.28±27.22,70.56±37.23 and 103.46 ± 46.62, respectively. There was a significant difference between the groups(F = 3.75, P < 0.05). Decreased expression of GST mRNA in fluorosis group and intervention group as compare to control was detected(all P < 0.05), and the expression of GST mRNA in intervention group was higher than that in fluorosis group(P < 0.05). Conclusion Coal-burning fluorosis possibly led to the decreased expression of GST mRNA in peripheral blood, and comprehensive control maybe prevent the decreased expression of GST in mRNA level.

Changes of the c-Jun N-terminal kinase in the brains of rats with chronicfluorosis / 中国地方病学杂志

Objective To investigate the expression of c-Jun-N-terminal kinase(JNK) in rat brains with chronic fluorosis and try to reveal the molecular mechanism for the neural impairment induced by the disease.Methods The rats were randomly divided into 3 groups, normal control group(drinking water containing less than 0.5 mg/L of sodium fluoride, NaF), lower fluoride exposed group(drinking water containing 5 mg/L NaF) and higher fluoride exposed group(drinking water containing 50 mg/L NaF), 24 in every group. The rats were examined at the sixth month after feeding. The concentration of fluorine in urine and blood was detected by F-ion selective electrode. The expression of JNK in brains was investigated by using Western blotting and immunohitochemistry staining, and analyze the correlation between activating of JNK and the concentration of fluorine in blood. Results The increased concentration of fluorine in urine(control: 0.92 ± 0.30, lower fluoride exposed group: 2.56 ± 0.91,higher fluoride exposed group: 5.73 ± 3.14, P ＜ 0.05) were observed when 6 months after the beginning of the experiment, and the amount of fluorine in blood was also higher in rats with fluorosis(control: 0.12 ± 0.07, lower fluoride exposed group: 0.36 ± 0.14, higher fluoride exposed group: 0.50 ± 0.18, P ＜ 0.05). The expression of phospho-JNK at protein levels were higher in the brains of rats with fluorosis than that of controls (control: 1.00 ± 0.37, lower fluoride exposed group: 1.20 ± 0.28, higher fluoride exposed group: 1.74 ± 0.69, P ＜ 0.05), whereas no change of total-JNK was found(F = 0.046, P ＞ 0.05). Furthermore, the expression of phospho-JNK in the parietal cortex(119.3 ± 14.1), occipital cortex(112.7 ± 5.4), hippocampus CA3(100.6 ± 8.9), dorsal thalamus (117.8 ± 10.4) and olivary nucleus( 112.6 ± 5.9) of rats in higher fluoride exposed group were higher than that in control( 104.1 ± 8.9,106.6 ± 9.6,106.6 ± 9.7,108.9 ± 6.4,100.3 ± 8.4, all P ＜ 0.05) and lower fluoride exposed group(96.7 ± 17.1,102.5 ± 8.3,106.4 ± 6.5,110.2 ± 9.3,102.4 ± 4.7,102.5 ± 9.8, all P＜ 0.05). The positive stained neurons of total-JNK also distributed in the same brain regions of rats, but no difference was detected between the rats with fluorosis and controls(all P ＞ 0.05). The increased level of phospho-JNK was positively correlated with the fluoride contents in blood of the rats with fluorosis (r = 0.677). Conclusions The expression of phospho-JNK in brains of rats with fluorosis was significantly increased with a correlation to fluoride content in blood, which might be connected to the mechanism of neural impairment induced by chronic fluorosis.

Blood routine test on the population in coal-burning fluoride toxicity in Bijie Country of Guizhou / 中国地方病学杂志

Objective To carry on a survey on blood routine examination of coal-burning endemic fluorosis population in Bijie City,Guizhou Province in order to study their health status and problems.Methods Blood routine examination was performed in the residents in coal-fired pollution endemic fluorosis-endemic area, including the residents of the Changchun Village of Changcun Town(intervention group)whose stoves had been improved and of Shiba Village Yachi Town not improved in Bijie City,Guizhou Province.The indicators were including leukocyte(WBC),red blood cell(RBC),hemoglobin(Hb),hematocrit(HCT),tlle average hematocrit red blood cell volume(MCV),mean corpuscular hemoglobin(MCH),mean corpuscular hemoglobin concentration (MCHC),red blood cell distribution width-CV(RDW-CV),platelets(PLT).Results RBC,Hb,HCT,MCHC, PLT were(4.95±1.18)×1012/L,(138.46±15.90)g/L,(50.19±11.48)%,(284.90±48.73)g/L,(334.92± 119.34)×109/L for the male in the intervened group,and they were(4.02±0.47)x 1012/L,(131.00±15.90)g/L, (40.90±7.60)%,(323.14±41.95)g/L,(280.79±100.34)× 109/L in non-intervention group,respectively. Inter-group comparison,the difference was statistically significant (U = 7.72,3.50,7.12,6.28,3.66,P ＜ 0.01). RBC, HCT,MCV,MCH,MCHC,RDW-CV,PLT were respectively(4.75±1.20)×1012/L,(46.91±11.20)%,(99.30± 6.88)fl,(28.10±8.66)pg,(275.61±54.49)g/L,(16.95±1.63)%,(351.23±150.37)×109/L for the female in the intervened group,and were (3.85±0.65)×1012/L,(38.80±6.60)%,(100.80±7.00)fl,(33.10±5.40)pg, (327.14±44.52 ) g/L,(16.60±1.58) %,(279.40±98.07)×109/L in the group un-intervened. Inter-group comparison found that there was a significant difference(U = 8.92,10.72,2.04,6.61,9.82,2.06,5.39,P ＜ 0.001 or 0.05) and the abnormal rate of RBC and Hb in non-intervention group[ 32.62% (92/282),16.67%(47/282)] was higher than that in the intervention group[9.73%(29/298) ,6.71%(20/298),x2 = 45.992,14.054,P ＜ 0.01 ) ]. Conclusion Experiment group has better results of blood routine test compared to non-intervention group,especially of anemia.

Changes in learning and memory ability and brain cholinesterase activity in the rats with coal burning fluorosis / 中国地方病学杂志

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.

Modified expression of extracellular signal-regulated protein kinase signal transducfion in rat braing and changed capacity of learning and memory of rats with chronic fluorosis / 中国地方病学杂志

Objective To investigate the expression of extraeellular signal-regulated protein kinase (ERK1/2)pathway in rat brains with fluorosis and the effects of fluoride on learning and memory of the rats,and to reveal the mechanisms of damaged nervous system resulted from the toxicity of the ion.Methods Seventy-two SD rats were divided into 3 groups and 24 rats were in each group.Three groups were fed respectively with different concentrations of fluoride(NaF)for 6 months to establish rat models with fluorosis.Controls were fed with tap water (NaF＜0.5 mg/L):lower and higher concentration group were fed with water containing NaF(5,50 ms/L).Animals are sacrificed after 6 months of treatment with fluoride and the dissected brains were kept for analysis.The protein levels of ERK1/2 in rat brains were detected by Western-blotting and the mRNA level by RT-PCR. The spatial learning and memorizing ability was measured by Morris water maze test. Results The ERK1/2 protein in control group,lower and higher concentration group was 0.944±0.10,1.253±0.02,1.953±0.07,the differece being statistieally sighificant between any two groups (P ＜ 0.05). The phospho-ERKl/2 protein in control group,lower and higher concentration group was 0.73±0.08,0.77±0.07,1.28±0.11,the differece being statistieally sighificant between any two groups(P ＜ 0.05);the activation rate of phospho-ERK1/2 in lower and higher concentration group [(68.4± 3.8)%,(64.1±3.2)%] was decreased compared to control group[ (82.3±10.7)%],the differece being significant(P ＜ 0.05). In the navigation trial,longer escape latencies of lower concentration group on the second, the third,the fifth and the sixth day were observed[ (46.0±8.0),(24.0±2.7),(8.9±5.3),(7.4±4.1 )s] compared to the control[ (39.3±6.9),(19.1±9.1 ),(8.3±3.4),(4.8±2.7)s],the differece being significant (P ＜ 0.05 or ＜ 0.01 );the similar results were also observed in the higher concentration group[ (36.9±16.8),(37.7±12.9), (19.7±7.6),(12.2±5.7 )s],and the escape latencies of the higher concentration group on the third,the fifth and the sixth day were longer than that in lower concentration group. In the probe test,the rats took more time to reach the first cross in lower and higher concentration group[(1.17±0.75),(4.18±1.10)s] than control group[ (5.89± 0.56 ) s ],the differece being significant (P ＜ 0.05 or ＜ 0.01 ) ;stayed shorter [ ( 17.05±4.25 ),(18.20±4.57 ) s ] than control [(25.37±5.65 )s ] in platform area (P ＜ 0.01 );the activation rates of ERK1/2 were directly correlated with the time taken to reach the first cross platform located in the probe test(r = 0.364,P ＜ 0.05) and the activation rates were also directly correlated with the escape latencies on the sixth day(r = 0.497,P ＜ 0.05). Conclusion Long-term exposure of excessive fluoride induces the change of expression and activating rate of the ERK1/2 in rat brains,leading to the decreased capacity of learning and memory.

Analysis of the GSTP1 gene Ile105Val polymorphism in fluorosis areas in Guizhou Province / 中国地方病学杂志

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.

Level of oxidative stress in rat brains and learning and memory function of rats with chronic fluorosis / 中国地方病学杂志

Objective To investigate the changes of oxidative stress level in brain tissues and serum, and learning and memory in rats with oxidative stress level in nerve damage in chronic fluorosis. Methods The rats were randomly divided into 3 groups according to the body weight, eight rats in each group, i.e., control group, drinking water containing less than 0.5 mg/L of fluoride; lower fluoride exposure group, drinking water containing 5 mg/L of fluoride; higher fluoride exposure group, drinking water containing 50 mg/L of fluoride. The animals were examined six months after initiating the experiment. The total antioxidant capacity (T-AOC) and malondialdehyde (MDA), as well as learning and memory, were measured. Results Escape latency in higher fluoride exposed group[ (14.37±3.48)s] was significantly higher than that of controls[ (5.84±1.87)s] and exposed te lower fluoride [ (7.18±1.42)s], the difference being statistically signifieant(P<0.05). As compared with controls[ (2.17±0.11)× 103 U/L , (0.79±0.11)×103 U/g Pr] ,the rats exposed to higher fluoride and lower fluoride exhibited lower levels of T-AOC [(1.37±0.27)×103 U/L,(0.24±0.06)×103 U/g Prand (1.20±0.14) x 103 U/L,(0.41 ~ 0.10)×103 U/g Pr], the difference being statistically signifieant(P<0.05). As compared with controls[ (2.34±0.16) mmoL/L, (2.97±0.11)mmol/g Pr] and low fluoride exposed group[ (2.68±0.33)mmoL/L, (3.38±0.21)mmol/g Pr], higher level of MDA were observed in higher fluoride exposed group[ (3.72±0.59)retool/L, (4.01±0.21)mmol/g Pr], the difference being statistically significant(P<0.05). Conclusion The results indicated that higher amount of fluoride induced an increased level of oxidation, which might result in the decreased capacity of intelligence of rats with fluorosis.

Study on the association of IL-10-592 polymorphism with susceptibility to hepatitis B viral infection in Han, Yi and Yao ethnic groups in Guizhou province / 中华流行病学杂志

Objective To investigate the association of IL-10 gene promoter polymorphism with susceptibility to hepatitis B viral infection in Han, Yi and Yao ethnic groups from Guizhou province.Methods Five hundred volunteers from Guizhou province were selected to undertake PCR-RFLP for detection of IL-10 gene promoter -592 polymorphisim. Results The genotypic distributions of IL-10-592 were 32.53%- 51.43% (AA),40.74%-54.82% (AC),5.79%-18.52% (CC) whereas the allelic frequencies were 59.94% -72.38% for the A allele, and 27.62%-40.06% for the C allele in Han, Yi and Yao ethnics from Guizhou. The distributions of allele and genotype frequencies of IL-10-592 were statistically different between Yao ethnic in Libo and Yi ethnic in Qianxi, Yao ethnic in Libo and Han ethnic in Libo, Yi ethnic in Qianxi and Yi ethnic in Weining, Yi ethnic in Weining and Han ethnic in Libo (P ＜0.05). IL-10-592 polymorphism was associated with HBV infection in Yi ethnic in Qianxi and the whole population. Conclusion IL-10-592 gene polymorphisms influenced the susceptibility to HBV infection in Han, Yao, Yi sub-populations in Guizhou. Result of the study suggested that IL-10-592 gene polymorphisms might serve as a risk factor to HBV infection.

Research into methylation of p16 gene in the patients suffering from coal-burnt arsenism in Xingren County Guizhou Province / 中华预防医学杂志

<p><b>OBJECTIVE</b>To probe into the situation and significance of p16 gene CPG island methylation in patients with arseniasis caused by coal-burning pollution.</p><p><b>METHODS</b>DNA was extracted using the Phenol-Chloroform method from leukocytes of 51 patients suffered from coal-burnt arsenism and 52 healthy volunteers. The quantity of the DNA was determined by UV spectrophotometry. Target DNA was denatured by NaOH, then the single strand DNA was modified by sodium bisulfite, converting all unmethylated (but not the methylated) cytosines to uracil. Subsequently a nested amplification with primers specific for methylated versus unmethylated DNA was performed, and PCR products were detected by gel electrophoresis.</p><p><b>RESULTS</b>Hypermethylation of the p16 CPG island was presented in 94.1% of the patients suffering from coal-burnt arsenism and in 73.1% of the healthy volunteers. There was statistical difference (P < 0.05) between them.</p><p><b>CONCLUSIONS</b>Methylation of p16 gene CPG island should have important pertinence in the metabolism of coal-burnt arsenism.</p>

Frequencies of GSTM1, GSTT1, and GSTP1 polymorphisms in ethnic minority from Guizhou / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the frequencies of GSTM1, GSTT1 and GSTP1 polymorphisms in Dong, Yi and Yao ethnic groups from Guizhou.</p><p><b>METHODS</b>In 321 volunteers who were population-based, GSTM1 and GSTT1 polymorphisms were analyzed by a multiplex-PCR procedure, whereas GSTP1 polymorphism was analyzed by PCR-RFLP method.</p><p><b>RESULTS</b>Null genotype for GSTM1 and GSTT1 was 59.6%-71.2% and 39.4%-72.5%, respectively. The genotypic distribution of GSTP1 was 63.3%-75% for AA, 23.2%-35.8% for AG, 0-1.9% for GG, whereas the allelic frequencies were 81.2%-86.6% for the A allele, and 13.4%-18.8% for the G allele.</p><p><b>CONCLUSION</b>There is a significant relationship between GSTT1 frequencies and ethnic populations.</p>

G6PD Gene Mutations in Shui people in Sandu of Guizhou / 中国实验血液学杂志

<p><b>UNLABELLED</b>Glucose-6-phosphate dehydrogenase (G6PD) deficiency is the most common human enzymopathy. To date, about 126 mutations in the G6PD gene have been detected, among which 17 mutations were found in Chinese. The most common mutations are: 1376 G-->T and 1388 G-->A, both in exon 12; 95 A-->G in exon 2, which amounted to more than 50% of mutations representing various regions and ethnic groups in China. A large-scale screening and genotypic analysis was held in Shui people in Sandu of Guizhou. To investigate the incidence and the molecular basis of G6PD deficiency of Guizhou Shui people, NBT qualitative and G6PD/6PGD quantitative methods were used to detect G6PD deficiency in 1,090 Shui people from the general people belonging to Sandu of Guizhou. By means of mis-matched primers amplified the G6PD gene, the products were 234 bp, 280 bp and 345 bp in length, then restriction enzyme analysis was used to detect the most common Chinese G6PD mutations, 1376 G-->T, 1388 G-->A and 95 A-->G. The results showed that out of the 1,090 samples, 98 G6PD deficiency samples were found. The incidence of G6PD deficiency was 8.99%. 24 cases of 1376 G-->T, 12 cases of 1388 G-->A, 9 cases of 95 A-->G were detected. A sample with 1376 G-->T and 95 A-->G mutation was found in a girl. It was reported for the first time.</p><p><b>IN CONCLUSION</b>1376 G-->T, 1388 G-->A, 95 A-->G mutations are the common G6PD mutations in Shui people in Sandu of Guizhou. The results indicates that different national minorities of Chinese may originated from a common ancestor.</p>