Table of Content

    05 April 2015, Volume 35 Issue 4
    Analysis of the screening sesult of cervical lesions of 24817 women undergone health examination in Hunan province
    2015, 35(4):  435-438. 
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    Objective To investigate the health state of the cervical of women in Hunan by gynecological general survey and to explore the best method of gynecological examination. Method:We analysed the result of 24817 Chinese women from Hunan Province who underwent gynecological examination in the examnation center of the third affliated Xiangya Hospital from July 2012 to July 2013.The examination included physical examination,cervical cytology(Thinprep cytologic test or Pap smear) and high-risk human papmomavirus(HR—HPV).Women with HPVpositive or aberrant TCT were advised to do colposcopy and histopathological examination. Results In all 24817 subjects,there were 14779 cases having chronic cervicitis (incidence:59.6%)。The sensitivity of TCT for screening cervical precancerous lesion or early cervical cancer is 92.56%,and .specificity is 80.05% , and thepositive rate was 2.95%,and the false-negative rate was 0.18%.And that of Pap smear was 57.35%,97.33%,1.65%,1.45%,respectively. There were significant difference between the two groups(P<0.05)。the rate of HR—HPV infection was 17.67%.Totally 709 biopsy specimens were obtained,46 of 709 patients were diagnosed CINI,36 were diagnosed CIN II,48 were diagnosed CINⅢ,10 patients were diagnosed early cervical cancer and 1 was diagnosed invasive cervical cancer。Using pathologic histology result as a gold standard, the incidence of CIN(31.4%)in populations with aberrant TCT results was higher than that in the normal populations (4.8%) (P<0.05),and also was much higher than the population with HPV-DNA positive(19.47%)(P<0.05). Conclusion The combined test of TCT and PHV contributes greatly to the early diagnosis of CIN,and TCT combined with HPV is the optial choice for screening of early cervical lssion and cervical cancer.
    Impact of serum high abundant protein immunodepletion on tissue proteome identification
    2015, 35(4):  439-443. 
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    Objective To analyze the impact of serum high abundant proteins immunodepletion on identification of tissue proteome. Methods Human thyroid and pituitary samples were disposed using 12 high abundant proteins depletion antibody column. The low abundant proteins and raw samples were analyzed by LC/MS. Identified peptides and proteins were qualitatively and quantitatively analyzed. Results The serum high abundant proteins in thyroid and pituitary were respectively depleted by 92.11%±12.87%, 78%±25%. Compared with raw thyroid, the non-redundant peptide and protein identification of low abundant thyroid was respectively increased by 70%, 16.5%. The non-redundant peptide identification of low abundant pituitary was 21.9% higher than raw samples. But the protein identification was unchanged. Conclusions The immunodepletion of serum high abundant proteins can significantly increase peptide and protein identification and is more suitable for the tissues with intensive blood vessel.
    Association between COMT Val158Met polymorphism and blood pressure and lipid levels in long-lived families
    2015, 35(4):  444-449. 
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    Objective To see the possible correlation between the COMT Val158Met polymorphism and blood pressure and serum lipid levels in a family-based long-lived population. Methods: COMT Val158Met was genotyped with PCR-RFLP for long-lived families (HL) and non-long-lived families (HNL) from Guangxi Hongshuihe River Basin and two controls outside the Basin (Pingguo long-lived families [PL] and non-long-lived families [PNL]). The correlation between this polymorphism and blood pressure and lipids were then evaluated. Results: No difference was found in the frequencies of allele A and its relevant genotypes (GA/AA) between HL and HNL, whose however are significantly higher than that of PL and PNL (all P < 0.01). While the blood pressure and lipid levels were similar between GA/AA and GG in HL and PNL, the systolic pressure, pulse pressure and levels of TC, TG and LDL-C of GA/AA were noted higher than were GG in HNL. Multiple regression analysis showed that COMT Val158Met was negatively associated with TC and LDL-C levels in HNL and PL. Conclusions: COMT 158Met mainly impact on blood pressure in the general families rather than the long-lived families in Guangxi Hongshuihe River Basin, albeit it presents more frequently in the Basin.
    GP78 gene polymorphism is associated with coronary heart disease
    2015, 35(4):  450-453. 
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    Objective To evaluate potential association between human GP78 gene and coronary heart disease(CHD). Methods The genotype was analyzed with by a Real-time PCR instrument in CHD group ( n = 557) and control group ( n=552).The data were evaluated via case control study. Results The distribution of rs2617849 genotype and allele frequency showed significant difference between CHD and control subjects( P < 0. 05) ,the distribution of the TT genotype and T allele was significantly higher in CHD patients than control subjects( P < 0. 05) .The significance of the TT genotype between CHD patients and control subjects retained after adjustment for covariates ( 95% CI: 1.035 ~1.736,P <0. 05 ). Conclusion rs2617849 polymorphism is closely associated with CHD,TT genotype and T allele of rs2617849 functions as a genetic markers of CHD.
    Application and investigation of a new genetic technology for Fragile X syndrome
    2015, 35(4):  454-457. 
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    Objective To establish a rapid、correct、available detection method for fragile X syndrome. Methods we used AmplideXTM FMR1 PCR Kit to detect samples from 26 suspected patients which included by 21 male and 5 female patients. All patients were also studied by using genomic EagI restriction enzyme PCR method in comparison with the above method. Result The results showed that the (CGG) n trinucleotide repeat of normal samples could be identified accurately by the PCR method. The results show there are 5 positive patients of fragile X syndrome which included by 4 full mutation patients and 1 female carrier. meanwhile, the results show 4 positive men patients, not detect the female carrier. Inclusion TP-PCR is a accurate, rapid, simple and inexpensive method. It could be used as a technology for screening fragile X syndrome.
    Expression of miR-574-5p in tissue with cervical cancer and effects of down-regulation of miR-574-5p on SiHa cell
    2015, 35(4):  458-462. 
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    Objective Investigate the expression level of tissue microRNA-574-5p (miR-574-5p) and the effects of down-regulation of miR-574-5p on SiHa cell; Methods The level of tissue miR-574-5p in 80 samples cervical cancer and 60 normal samples as controls were detected by real-time RT-PCR. Transfected the miR-574-5p inhibitor into SiHa cell, and detected the proliferation, the apoptotic rate and the migration of SiHa cell through MTT, the flow cytometer and transwell experiment respectively. Results Expression levels of tissue miR-574-5p was significantly higher in cervical cancer than that in normal controls(P<0.001); Up-regulation expression of miR-574-5p had a significantly positive correlation with the International Federation of Gynecology and Obstetrics (FIGO) stage and pathological grade and lymph node metastasis (P<0.05 ), while it had no clear-cut correlations with the age of patients and menopausal status. After transfected the miR-574-5p inhibitor into SiHa cell, the proliferation and the migration were decline(P<0.05), and the apoptotic rate was addition(P<0.05). Conclusion The high expression of miR-574-5p in the tissue of cervical cancer may be related to the progress in the malignant cervical cancer, which expected to be a potential targeting therapy of cervical cancer through the effects of transfecting miR-574-5p inhibitor on SiHa cell.
    Phorbol myristate acetate promotes differentiation of mouse induced pluripotent stem cells into cardiomyocytes in vitro
    2015, 35(4):  463-469. 
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    Objective To investigate the effect of PMA on the differentiation of mouse induced pluripotent stem cells into cardiomyocytes in vitro, and to establish an efficient and safe protocol for cardiac differentiation in vitro. Methods In this experiment, embryoid bodies (EBs) were formed by classical hanging drop method, and were directional differentiated to cardiomyocyte with PMA treatment; Furthermore, expression of cardiac troponin T and α-Sarcomeric actinin was observed by immunofluorescent staining, mRNA expression levels of the related genes Brachyury, cTnT, MLC2a, NKX2.5, GATA4 were analyzed by qPCR and RT-PCR.Control group was treated by DMSO, number of beating embryoid bodies was calculated. Results The best concentration of PMA to induce the cardiac differentiation of miPSC was 100 nmol/L and the beating area was found in 53% of embryoid bodies, compared with the control group (beating area was found in 12% of embryoid bodies), Differentiation ef?ciency was higher in PMA treatment group. IPS-CM expressed serve cardic related proteins and genes, and had the Structure characteristics of myocardium cell. Conclusion: PMA can promote miPSC to differentiate into cardiomyocyte in vitro.
    Effects of hypoxia on the expression of miR-124 and BACE1 protein in SH-SY5Y cells
    2015, 35(4):  470-474. 
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    Objective To investigate the role of hypoxia on the expression of miR-124 and BACE1. Methods Cultured SH-SY5Y cells were divided into blank control group, oxygen-glucose deprivation (OGD) group, Aβ1-42 group and overexpression or inhibition of miR-124 group. Real-time quantitative PCR was performed to analyze the expression of miR-124 level. The level of BACE1 was examined by Western blot. Results After OGD for 48h, the expression level of miR-124 decreased significantly to 46.9% of the control group (P<0.05), while the expression level of BACE1 was increased by 36% compared with the control group (P<0.01). After transfection of 48h, miR-124 level of miR-124 mimic group was increased by 245 folds as compared with control (P<0.01) and by 219 folds as compared with miR-124 mimic control (P<0.01); BACE1 level was decreased by 29% compared with control (P<0.05) and by 25% compared with miR-124 mimic control (P<0.05); miR-124 level of miR-124 inhibitor group was decreased to 45.4% of control (P<0.05), 48% of miR-124 inhibitor control (P<0.05), BACE1 level was increased by 21% compared with control (P<0.05), 40.3% compared with miR-124 inhibitor control (P<0.01). miR-124 level was decreased by 52% compared with control (P<0.05) and the level of BACE1 level was increased by 51% after Aβ1-42 treatment fo