30%Horseradish Peroxidase was ingected into the nueleus lateralis dorsalis and nucleus lateralis posterior of thalamus. The result shows that thalamus has much more labeled cells than the cerebral. The midbrain has the leaet.We discussed the labeled cells of the cerebral cortex, putamencaudatus, nucleus reticular and ventral nucleus of thalamus, formatio retieaularis of midbrain.

Objective The frequency of G6PD deficiency in the Tujia nationality in Jiangkou,Guizhou was investigated.Methods In Oct.2002,227 male subjects were selected randomly from the local people,and NBT qualitative and G6PD/6PGD quantitative methods were used to detect G6PD deficiency in them.Results Among the 227 subjects,17 cases of G6PD deficiency were found.The gene frequency of G6PD deficiency was 0.0749.Conclusion There is a high incidence of G6PD deficiency in Tujia nationality in Jiangkou,Guizhou.The investigation will illustrate the distribution of G6PD deficiency in Guizhou,and provide some useful data for the preventing G6PD deficiency,directing clinical management,improving minority population diathesis and studying the origin of Tujia nationality.

Objective To compare the transfection efficiency between different transfection methods in human HepG2 and SGC7901/ADM cells so as to provide experimental basis for further study .Methods To electrons fect the enhanced GFP plasmid into HepG2 and SGC7901/ADM cells by lipofection and electroporation methods ,respectively .The survival rates and transfection efficiency were analyzed .Results The efficiency of eGFP vector transfected into HepG2 cells by lipofection was (23 .8 ± 2 .1)% , compared with lipofection method ,the efficiency of eGFP plasmid transfected by electroporation was up to (49 .6 ± 2 .5)% ,and the difference was statistically significant(P<0 .05) .The efficiency of SGC7901/ADM cells by lipofection was (25 .4 ± 1 .3)% ,com‐pared with lipofection method ,the efficiency of electroporation was up to(52 .6 ± 2 .1)% ,and the difference was statistically signifi‐cant(P<0 .05) .This study provides reliable test parameters for electransfection of HepG2 and SGC7901/ADM cells .Conclusion The transfection efficiency of large fragment vector is efficiently improved by electroporation .

OBJECTIVE To study the distributive characteristics of the pathogens of the bacterial infectious diseases in Wuzhou,and to provide the basis for prevention and therapy of the infectious diseases.METHODS The VITEK-ATB system,BDPhoenix100 system,API system and CHROMagar were used to appraise the pathogens,which were from the bring up of doubtful infectious disease specimens.RESULTS The positive rate of 4 878 pathogenic test was 32.10%.The Gram-negative bacilli were the major(63.96%) from the 1 623 strains of pathogens isolated.The main composition of pathogen spectrum was Escherichia coli,Klebsiella pneumoniae,Staphylococcus aureus,Pseudomonas aeruginosa,Acinetobacter baumannii,Candida albicans,Enterococcus faecalis,Staphylococcus haemolyticus,Enterobacter cloacae,and Staphylococcus epidermidis.The distribution of main pathogens had different characteristics in different season and different infectious diseases.CONCLUSIONS Strengthening the test of pathogens in the doubtful infectious diseases and understanding the characteristics of local pathogen spectrum and the transitional current of infections in time,significantly influence on prevention and therapy of the infectious diseases.

Objective To investigate the correlation between fibroblast growth factor receptor 2 (FGFR2) gene polymorphism and endemic fluorosis.Methods In Bijie City,Guizhou Province coal-burning-borne high fluoride areas,148 patients with fluorosis were selected as endemic fluorosis group;in non high fluoride areas of Changshun County of Guizhou Province,134 healthy people were selected as control group.Short tandem repeats (STRs)-PCR was utilized to detected the FGFR2 rs35668561 and D10S14839 microsatellite polymorphisms in endemic fluorosis cases and controls.Results FGFR2 rs35668561 461 bp (22AG)allele frequency of endemic fluorosis group (1.01％) was significantly lower than that of the control group (3.36％,x2 =5.29,P ＜ 0.05).FGFR2 D10S14839 286 bp (9GT),300 bp (16GT),310 bp (21GT) and 314 bp (23GT) allele frequency in the endemic fluorosis group were 14.53％,11.82％,16.89％ and 8.11％,in the control group were 22.01％,6.34％,8.96％ and 16.42％,the difference was statistically significant.Then 300 bp (16GT)and 310 bp (21GT)allele frequency of endemic fluorosis group was significantly higher than that of the control group (x2 =6.82,7.77,all P ＜ 0.05),and 286 bp (9GT),314 bp (23GT) allele frequency of endemic fluorosis group was significantly lower than that of the control group (x2 =5.32,9.16,all P ＜ 0.05).Conclusions FGFR2 rs35668561 and D10S14839 polymorphism are associated with endemic fluorosis.FGFR2 rs35668561 461 bp (22AG) allele may be a protective factor of endemic fluorosis.D10S14839 300 bp (16GT) and 310 bp (21GT) allele may be risk factors of endemic fluorosis,286 bp (9GT) and 314 bp (23GT) allele may be protective factors of endemic fluorosis.

Objective To explore the correlation between myeloperoxidase (MPO) genetic variation and coal-burning endemic fluorosis, and to understand the influence of integrated intervention including stove changes and health education on people’s health in the area. Methods In 2007, coal-burning endemic fluorosis disease areas were selected in Bijie City, Guizhou Province. No stove changes in Yachi Town, 150 patients with dental fluorosis were selected as fluorosis non-intervention group, and the intervention group was 150 patients in Changchun Town where the stoves were changed 2 years ago. The population in control group was selected in an area with non-endemic fluorosis in Changshun County. The mRNA expressions of MPO in leukoxytes were detected by real-time PCR. HepG2 cells were cultured in vitro and divided into four groups: pGL3-A group, pGL3-G group, pGL3-Control group and pGL3-Basic group. pGL3-A and pGL3-G were recombinant plasmid, while pGL3-Basic as a blank control and pGL3-Control as a positive one. The internal reference plasmid pRL-TK co-transfected the HepG2 cells with pGL3-G, pGL3-A, pGL3-Basic and pGL3-Control, respectively. The influence of sudden change of MPO gene promoter on the gene transfection activity was evaluated by a dual luciferasereporter gene system. Results The expression level of MPO mRNA in peripheral blood leukocytes in non-intervention group(0.054 ± 0 . 003 ) were higher than control and intervention groups (0.019 ± 0.004,0.019 ± 0.003, all P<0.05), and no significant change was found between intervention group and control group(P>0.05). After the MPO-463G/A locus genetic variation occured, the luciferase reporter gene expression level of the recombinant plasmid pGL3-G(0.753 4 ± 0.086 6) was higher than that of the pGL3-A(0.490 0 ± 0.022 3, P < 0.05). Conclusions The study on MPO gene promoter-463G/A locus has prompted that MPO gene allele may be a protective factor to coal-burning fluorosis. The integrated interventions have a role in the prevention and treatment of endemic fluorosis.

Objective:To investigate the association between polymorphisms of FGFR2 and the susceptibility of breast cancer in Han population in Guizhou province.Methods:Genotyping was performed using PCR-sequence-specific primers(PCR-SSP)in 106 histologically confirmed breast cancer cases and 116 cancer-free controls.Results:The genotype frequencies of rs1219648 TT,TC,and CC were 50%,25.47%.and 24.53% in breast cancer cases and 29.31%,48.28%,and 22.41% in the controls.The gene frequencies of T in breast cancer cases and the controls were 62.74% and 53.45%.respectively.The gene frequencies of C were 37.26% and 46.55%.respectively.The distribution of allele and genotype frequencies of FGFR2 rs1219648 was statistically different between breast cancer cases and the controls(P<0.05).Conclusion:FGFR2 rs1219648 polymorphism influences the susceptibility of breast cancer.TT genotype might serve as a risk factor for breast cancer.

Objective In the present study, we investigated the effects of advanced oxidation protein products(AOPP) on reactive oxygen species(ROS) production in murine osteoblastic MC3T3-E1 cells by NADPH oxidase enzymes pathway. Methods Experiments were divided into three groups, including control group, rats albumin(RSA) group, and AOPP group. Different concentrations of AOPP were added to the osteoblastic MC3T3-E1 cells culture medium. The production of ROS in MC3T3-E1 cells was measured by the fluorescence intensity of intracellular fluoroprobe ( DCFD ) . In order to verify the effect of enzyme of the production of ROS, the specific inhibitors of corresponding enzymes were added in the MC3T3-E1 cells which were cultured in the medium with AOPP. Finally, western blot and immunofluorescence were used to observe the changes of NADPH oxidase enzymes subunits. Results Different concentrations of AOPP (50,100,200μg/ml) induced MC3T3-E1 cells to produce different amount of ROS. The higher concentrations of AOPP were added, the more ROS were produced. Furthermore,200μg/ml AOPP induced the maximum amount of ROS production(P<0. 05). Meanwhile, AOPP induced MC3T3-E1 cells to produce different amount of ROS with a time-dependent manner. The peak amount of ROS production in MC3T3-E1 cells was observed in 3h when AOPP were added (P<0. 05). In addition, when specific inhibitors of corresponding enzymes were added in the MC3T3-E1 cells, the production of ROS were significantly suppressed by C-SOD, DPI, and apocynin(P<0. 05). On the other hand, AOPP can up-regulate the expression of Nox4 protein of the MC3T3-E1 cells, which is one of the subunits of NADPH oxidase enzymes. Meanwhile, AOPP can also induce the membrane migration of p47phox subunit. Conclusion AOPP induces osteoblastic MC3T3-E1 cells to produce ROS by NADPH oxidase enzymes pathway, and which may be one of the pathogenesis of AOPP involved in osteoporosis.

OBJECTIVETo analyze the population genetics characteristics of mitochondrial DNA (mtDNA) in Gelao, Mulao, Maonan ethnic groups from Guizhou.

METHODSMinisequenceing and restriction fragment length polymorphism (RFLP) were used to analyze 12 single nucleotide polymorphism (SNPs) of mitochondrial DNA in the 3 ethnic groups.

RESULTSA total of 30 haplotypes were detected in 156 samples. The distribution of H1, H23 had differed between Mulao, Maonan and Gelao, respectively, and so did M7 among the three groups. The difference was statistically significant (P < 0.05). Mulao, Maonan had respectively differed from Gelao and the difference was also statistically significant (P < 0.05).

CONCLUSIONThere was a great similarity in the distribution of haplotypes of the mtDNA among the three ethnic groups, except for some difference in the distribution of certain haplotypes.

Asian Continental Ancestry Group ; ethnology ; genetics ; China ; ethnology ; DNA, Mitochondrial ; genetics ; Humans ; Male ; Pedigree ; Polymorphism, Genetic

BACKGROUNDHuman cytomegalovirus (HCMV) infection is an important infectious agent that results in neonatal disease and congenital deformity. HCMV infection may affect in many organs. The different symptoms and tissue tropism of HCMV infection perhaps resulted from the genetic polymorphism of HCMV. HCMV UL144 open reading frames encode a homologue of the tumor necrosis factor receptor. It seems important to study the strain-specific variability of UL144 sequence in low-passage clinical isolates and to discuss if the variability related to the clinical HCMV infection.

METHODSHCMV-UL144 gene was amplified by PCR assay in 65 low-passage clinical isolates and urine from 7 healthy children who were HCMV-DNA positive by quantitative PCR. All the positive PCR products were analyzed by Heteroduplex mobility assay and single-stranded conformation polymorphism (HMA-SSCP) and 32 of them were sequenced.

RESULTSFifty-five isolates and 5 urine specimens were HCMV-UL144 positive by PCR. Sequencing and HMA-SSCP analysis showed that significant strain-specific variability was present in the UL144 ORFs. Comparing UL144 sequences and the corresponding symptoms showed that genotype 2 did not exist in megacolon isolates. And genotype 1 and 3 were the major types among microcephaly and jaundice isolates respectively.

CONCLUSIONHCMV-UL144 existed in almost all the low passage isolates. HMA-SSCP assay is an easy and effective method to detect the genetype of HCMV-UL144 sequence. The characteristic of sequences in different isolates showed that UL144 gene may play an important role in HCMV infection.

Cytomegalovirus ; classification ; genetics ; isolation & purification ; Cytomegalovirus Infections ; virology ; Genotype ; Humans ; Infant ; Infant, Newborn ; Membrane Glycoproteins ; genetics ; Molecular Sequence Data ; Phylogeny ; Polymerase Chain Reaction ; Polymorphism, Single-Stranded Conformational ; Sequence Analysis, DNA ; Viral Proteins ; genetics