Objective To identify the distribution feature of methylenetetrahydrofolate reductase (M T HFR) gene polymorphism of Buyi ,Dong ,Miao nationality in Guizhou .Methods The MTHFR(677 and 1 298) genotypes of Buyi ,Miao and Dong healthy indi-viduals were determined by TaqMan-MGB probe genotyping method and constructed haplotypes .Results There were significant difference of MTHFR 677C/T genotype and allele frequencies among 3 groups(P<0 .05) ,There was significant difference of geno-type between Buyi and Miao nationality ,and there were significant differences of genotype frequencies in Buyi nationality and Dong and Miao nationality(P<0 .01) .There were no differences of MTHFR 1298A/C genotype frequencies among Buyi ,Dong and Miao nationality(P> 0 .05) .Buyi nationality had the lowest frequency in double wild homozygous type (677CC/1298AA) ,677TT/1298CC double mutation homozygous and 677TT/1298AC combination in above three minorities was not found .There were linkage disequilibrium between 677C/T and 1298A/C in Buyi and Miao nationality .Conclusion The genotypes frequencies of MTHFR 677T T/1298AC are significant differences among different regions and different ethnic groups .

Objective To investigate the effect of fluoride on the expression of a subunit of calcium/calmodulin-dependent protein kinase- Ⅱ (α-CaMK Ⅱ ) at both mRNA and protein levels in human neuroblastoma cells were cultured in DMEM with final concentrations of NaF 0(control) ,0.05,0.50,2.00,5.00 mmol/L, respectively, for 48 hours. Then quantitative RT-PCR and Western blot were performed to detect the expression level of α-CaMK Ⅱ P1 (postnatal day 1) pups together with their mothers were randomly divided into three groups. Lactating rats were given drinking water containing NaF at concentrations 0(control) ,2,3 mmol/L. And pups were exposed to NaF through milk. In each group, 8 pups were sacrificed on day 14 after birth. In post-weaning period, another 8 pups in each group were given drinking water with the same dose of fluoride as their mother's 21 day after birth. After then, these pups were killed on day 28, and hippocampus was dissected immediately and Western blot was conducted mRNA and protein levels were decreased. When NaF concentrations were 0,0.05,0.50,2.00,5.00 mmol/L, the mRNA relative ratios of α-CaMKⅡ in SY5Y cells were 1.00 ± 0.00,0.77 ± 0.18,0.40 ± 0.11,0.22 ± 0.06 and 0.15 ± 0.03, and protein levels of α-CaMK Ⅱ were 100.00 ± 0.00,76.17 ± 2.08,59.16 ± 2.12,48.52 ± 2.71 and 43.51 ± 2.57, any mmol/L group, hippocampus α-CaMK Ⅱ protein levels on day 14 and 28(75.02 ± 2.88,73.83 ± 3.88 and 81.00 ± 2.54,45.70 ± 2.34) were significantly lower than that of control groups(100.00 ± 0.00,100.00 ± 0.00, all P < 0.01). In 3 mmol/L group, hippocampus α-CaMKⅡ protein level on day 28 was lower than that of 2 mmol/L group (P < 0.01). Conclusion Fluoride can decrease mRNA and protein levels of α-CaMK Ⅱ in nerve cells and hippocampus, which may be one of the mechanisms of learning and memory impairment by fluorosis.

Objective To investigate the clinical application of multiplex allele-specific PCR assays for simultaneous detection of the mitochondrial 12S rRNA A1555G and C1494T mutations associated with aminoglycoside-induced hearing impairment.Methods Three standard plasmids of different genotypes (wild-type, A1555G mutant and C1494T mutant) were constructed for templates and allele-specific primers aiming directly at wild-type and mutant of mitochondrial DNA nt1555 and nt1494 were designed for developing a multiplex allele-specific PCR technique to detect the A1555G and C1494T mutations.Then the method was applied to clinical screening of 138 non-syndromic hearing loss subjects and confirmed by DNA sequencing.Results Multiplex allele-specific PCR was successfully applied to the detection of A1555G and C1494T mutations in a cohort of 138 Han Chinese genetically unrelated hearing-loss subjects.Finally, 11(7.97%) unrelated affected subjects harbored the A1555G and C1494T mutations in the 12S rRNA gene(10 cases for A1555G and 1 cases for C1494T), which was well consistent with results of DNA sequencing [7.97%(11/138), Kappa=1.000, P＜0.01].Conclusion This study indicates that the multiplex allele-specific PCR assay is useful, convenient and reliable in the detection of the A1555G and C1494T mutations, which could identify the subjects at risk and effectively prevent of aminoglycoside-induced hearing loss.

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:This study aimed to investigate the effect of human lymphatic endothelial cells (HLECs) on proteins secreted by epithelial ovarian cancer (EOC) cells SKOV3-pm4 with highly directional lymphatic metastasis. Methods:The supernatants of the four groups of cultured cells (A, SKOV3;B, SKOV3+HLEC;C, SKOV3-PM4;and D, SKOV3-PM4+HLEC) were collected. The proteins of these cells were detected by antibody arrays and iTRAQ-2D-LC-MALDI-TOF/TOF/MS. The screened significantly differential proteins were further analyzed by bioinformatics and validated in the human serum and cell culture medium by ELISA. Results:Progranulin (GRN) and vascular endothelial growth factor A (VEGF-A) were upregulated between groups C and A. In addition, insulin-like growth factor binding protein-7 (IGFBP-7) and secreted protein acid rich in cysteine (SPARC) were downregulated between groups D and C. Comprehensive bioinformatics analysis revealed that IGFBP7 interacted with VEGFA. VEGF exhibited the highest expression in ovarian cancer and IGFBP7 exhibited the lowest expression compared with the serum of the normal control group. Statistically significant differences were observed between the two substances. Conclusion:The HLEC microenvironment is closely associated with directional metastasis in lymph nodes with differential proteins, including matricellular proteins and adhesion factors. In particular, the upregulation of VEGFA and GRN and the downregulation of SPARC and IGFBP7 were closely associated with the directional metastasis of EOC cells in lymph nodes.

Objective:To explore the effect of sarpogrelate on platelet function in patients at the bridging stage before coronary artery bypass grafting (CABG).
Methods: A total of 40 consecutive patients with peripheral artery stent and scheduled for CABG in our hospital from 2011-05 to 2013-04 were enrolled in this study. The patients were randomly divided into 2 groups, Low molecular weight heparin (LMWH) alone group, n=19 and Sarpogrelate+LMWH group, n=21. The medications started at 5-7 days before CABG and stopped at 24 h before CABG. The platelet inhibition rates (platelet aggregation induced by collagen+ serotonin) were examined and compared between 2 groups at the baseline (before randomization), 24h and 1h before CABG respectively.
Results: The platelet inhibition rates were similar between 2 groups at the baseline (87.33 ± 6.82) % vs (86.11 ± 6.87) %, P=0.577 and 1h before CABG (62.60 ± 12.39) % vs (56.19 ± 14.99) %, P=0.148. At 24h before CABG, the platelet inhibition rate in Sarpogrelate+LMWH group was higher than that in LMWH alone group (83.87 ± 8.99)%vs (63.13 ± 10.88)%, P<0.001. Compared with the baseline, the falling range of platelet inhibition was lower in Sarpogrelate+ LMWH group at 24h before CABG, (3.46 ± 6.18) % vs (22.98 ± 9.43) %, P<0.001 and the falling range was similar between 2 groups at 1h before CABG (24.73 ± 14.19)%vs (29.92 ± 14.28)%, P=0.257.
Conclusion: Sarpogrelate + LMWH may result better platelet inhibition rate with quicker recovery of platelet function upon the medication stopping, which might be a feasible management in patients at the bridging stage before CABG.

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 understand the ABO blood type distribution of Buyi and Shui ethnic populations in Libo county of Guizhou province .Methods Totally 726 Buyi and 163 Shui individuals who were unrelated within three generations were detected ABO blood stype with glass-clotted method .Results The ABO blood type distributions in the Buyi and Shui ethnic populations both were in line with the Hardy-Weinberg equilibrium .The Buyi ethnic population ABO phenotypic distribution and gene frequen-cies were O>B>A>AB and r>q>p (r=0 .676 9 ,q=0 .176 7 ,P=0 .146 4) and the national index was 0 .842;while the Shui eth-nic population ABO phenotypic distribution and gene frequencies were O>A>B>AB and r>p>q(r=0 .522 6 ,q=0 .104 0 ,P=0 .164 7) and the national index was 1 .529 .Conclusion The Buyi and Shui ethnic populations′ABO blood type distribution in Libo county of Guizhou province is basically consistent with that of the same ethnic populations in other regions of Guizhou Province of the same ethnic groups .The genetic distance analysis prompts that the regional and ethnic differences exist in ABO blood type dis-tribution .

Objective To investigate the effect of duodenal-jejunal bypass(DJB) on glucose metabolism and the expression of ileum proglucagon mRNA in Goto-Kakizaki (GK) type 2 diabetic rats.Methods 18 male GK rats and 1 8 male Wistar rats were randomly divided into 4 groups:GK operation group(group A),GK sham operation group(group B),Wistar operation group (group C),Wistar sham operation group (group D).There were 9 rats in each group.The fasting blood glucose and GLP-1 levels were measured before and at the 8th weekafter surgery.Oral glucose tolerance test was measured,and the area under blood glucose concentration curve wascalculated.Ileum tissues were obtained 8 weeks postoperatively and reverse transcriptase polymerase chain reac-tion was used to detect ileum PG mRNA expression after the operation.Results At the 8th week after surgery,the fasting blood glucose of group A decreased from(8.73 ± 1.30) mmol/L to(5.86 ±0.57) mmol/L(P ＜0.05).Area under blood glucose concentration curve decreased from preoperative(60.23 ± 5.14)mmol · h/L to (47.80 ±1.79) mmol· h/L(P ＜ 0.05).Fasting serum GLP-1 in group A increased from (7.69 ± 0.74) pmol/L to (29.00 ±4.85) pmol/L while postprandial GLP-1 increased from(15.74 ± 5.71) pmol/L to (45.78 ± 7.26) pmol/L (P ＜0.05).At the 8th week after surgery,the ileum PG mRNA level in group A was significantly higher than that in group B(P ＜ 0.05).Conclusions DJB can directly improve glucose metabolism in non-obese type 2 diabetic rats,and the operation has no hypoglycemic effect on normal blood glucose.The increased expression of ileum PG mRNA might contribute to the hypoglycemic effect of the surgery.

Objective To investigate the diagnostic value of texture analysis derived from conventional MR imaging in differentiating benign and malignant breast lesions. Methods Thirty-six patients with malignant breast lesion and 33 patients with benign breast lesion were retrospectively analyzed in our study. All patients underwent conventional MR imaging including axial T1WI, T2WI, and contrast-enhanced T1WI before surgery. Texture features were calculated from manually drawn ROIs by using MaZda software. The feature selection methods included mutual information (MI), Fishers coefficient, classification error probability combined with average correlation coefficients (POE + ACC) and the combination of the above three methods(FPM). These methods were used to identify the most significant texture features in discriminating benign breast lesion from malignant breast lesion. The statistical methods including raw data analysis (RDA), principal component analysis (PCA), linear discriminant analysis (LDA) and nonlinear discriminant analysis (NDA) were used to distinguish malignant breast lesion from benign breast lesion. The results were shown by misclassification rate. Results In the three kinds of sequences, the texture features for differentiating malignant breast lesion and benign breast lesion were mainly from T2WI which had the lowest misclassification rate 4.35%(3/69). The misclassification rates of the feature selection methods were similar in MI, Fisher coefficient and POE+ACC (15.94%to 56.52%for MI;17.39%to 56.52%for Fisher coefficient and 17.39%to 56.52%for POE+ACC). However, the misclassification rate of the combination of the three methods (4.35%to 53.62%for FPM) was lower than that of any other kind of method. In the statistical methods, NDA (4.35% to 27.54%) had lower misclassification rate than RDA (33.33% to 56.52%), PCA (33.33% to 53.62%) and LDA (15.94% to 44.93%). Conclusion Texture analysis of conventional MR imaging can provide reliably objective basis for differentiating benign from malignant breast lesions.