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Objective@#To investigate the role of human papillomavirus (HPV) 16 early genes E2 and E6 and heterogeneous nuclear ribonucleoprotein (hnRNP) E2 and their interaction effects in the progression of the cervical cancer.@*Methods@#Women with normal cervix (NC), low cervical intraepithelial neoplasia (CIN Ⅰ) and high cervical intraepithelial neoplasia (CIN Ⅱ/Ⅲ) from the cervical lesions cohort in Jiexiu County of Shanxi Province from June 2014 to September 2014, and patients with cervical squamous cell carcinoma (SCC) treated at the Second Hospital of Shanxi Medical University in the same period were enrolled in this study. There were 257 participants, about 67 NC cases (26.07%), 69 CIN Ⅰ cases (26.85%), 68 CIN Ⅱ/Ⅲ cases (26.46%), and 53 SCC cases (20.62%), respectively. The information of demographic characteristics, life health habits and cervical lesions were collected by using the structured questionnaire. Cervical exfoliated cells and cervical biopsy tissues were collected to detect the infection of HPV16 and the protein expression levels of hnRNP E2, HPV16 E2 and E6. According to the median-value of the protein expression levels of hnRNP E2, HPV16 E2 and E6 and E2/E6 ratio in the NC group, the study participants were divided into the high and low expression groups/ratio groups. The multivariate logistic regression model was used to analyze the correlation between HPV16 early gene E2 and E6, hnRNP E2 and cervical cancer. The interaction effect was analyzed by using the generalized multifactor dimensionality reduction (GMDR) model.@*Results@#The ages of NC, CIN Ⅰ, CIN Ⅱ/Ⅲ and SCC groups were (47.00±9.07), (47.64±7.35), (46.37±8.67) and (51.26±8.03) years old, respectively. The multivariate logistic regression model analysis showed that the HPV16 E2 low expression, E6 high expression and E2/E6 low ratio could increase the risk of CIN Ⅱ/Ⅲ, about OR (95%CI) values 11.11 (1.63-75.56), 8.00 (1.28-50.04), and 9.75 (1.22-77.72), respectively and SCC, about OR (95%CI) values 14.22 (2.11-95.88), 10.33 (1.67-64.00), and 12.38 (1.56-97.91), respectively. The hnRNP E2 low expression could increase the risk of CIN Ⅱ/Ⅲ and SCC, about OR (95%CI) values 3.35 (1.39-8.10) and 5.53 (1.54-19.88). The result of GMDR showed that there were interaction effects of the hnRNP E2 low expression, HPV16 E2 low expression and HPV16 E6 high expression in both CIN Ⅱ/Ⅲ and SCC groups.@*Conclusion@#The HPV16 E2 low expression, HPV16 E6 high expression and hnRNP E2 low expression could increase the risk of high-grade cervical intraepithelial neoplasia and cervical cancer, and they might have an important interaction effect in the progression of the cervical cancer.
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Objective To investigate the effect of heterogeneous nuclear ribonucleoprotein K (hnRNP K) and its interaction with human papillomavims 16 (HPV16) on cervical intraepithelial neoplasia (CIN).Methods The participants included 67 women with normal cervix (NC),69 women with CIN Ⅰ and 68 women with CIN Ⅱ/Ⅲ in a community cohort of pathologically diagnosed women established in Jiexiu of Shanxi province,from June 2014 to June 2015.A structured questionnaire was used to collect the demographic data of the subjects and the related factors of cervical lesions.Cervical exfoliated cells and cervical tissues from biopsy or surgery were selected.The infection status of HPV16 was detected by flow-through hybridization.The protein expression levels of hnRNP K were evaluated by Western blot.SPSS 23.0 software was used to collate and analyze the data.To study the differences in demographic characteristics,related factors,hnRNP K protein and HPV16 infection among NC,CIN Ⅰ and CIN Ⅱ / Ⅲ groups,X2 test,trend x2 test,and Kruskal-Wallis H test were conducted.Multiple comparisons of hnRNP K protein in three groups were completed by using the Bonferroni method.The OR and its 95％CI of hnRNP K,HPV16 and CIN were calculated by using the unconditional logistic regression models.Two-way interactions between hnRNP K protein and HPV16 infection on CIN were analyzed by using additive model and related indicators.Results HPV16 infection rates were 10.4％ in women with normal cervix,14.5％ in women with CIN Ⅰ and 41.2％ in women with CIN Ⅱ/Ⅲ,respectively.The differences among three groups were significant (P＜0.001).Moreover,the infection rates of HPV16 gradually increased with the increasing severity of CIN (trend x2=18.512,P＜0.001).The differences in protein expression of hnRNP K among three groups were significant (H=48.138,P＜0.001) and the expressionincreased with the development of cervical lesionss (trend x2=21.765,P＜0.001).Results from the interaction analysis indicated that there were additive effects between high expression of hnRNP K protein and HPV16 in CIN Ⅱ/Ⅲ group compared with normal group (API=0.639,95％CI:0.083-1.196).In contrast,no such additive effect was found in CIN Ⅰ group.Conclusions HPV16 infection and over-expression of hnRNP K protein were associated with the increased risk of cervical intraepithelial neoplasia.There might be interaction between hnRNP K protein overexpression and HPV16 infection existed on the progress of CIN Ⅰ/Ⅲ.
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Objective To explore the relationship between male sexual function and daily exposure to bisphenol A (BPA) at a reproductive center in Taiyuan.Methods Male patients who were seeking treatment of infertility due to problems caused by either of the spouse were selected from the Shanxi reproductive center between September 2014 and April 2015.Information on general characteristics,sexual function and fasting venous blood samples were collected.Total scores of sexual function were evaluated by Delphi expert scoring method.Levels of serum BPA were measured by high-performance liquid chromatography.Data was analyzed by Spearman rank correlation,rank sum test,multivariate logistic regression analysis andx2 trend test.Relationship between BPA and sexual function was presented as OR and corresponding 95％CI.Results Among the 353 participants,45.0％ was defined as sexual dysfunction with low sexual desire (47.3％) as the major reason.BPA was detected in all the 353 patients,with a range of concentration as 0.38-21.93 ng/ml and an average as 5.79 ng/ml.Results from the Spearman rank correlation analysis revealed significant negative correlations between serum BPA and sexual function,sexual desire,erectile ability and ejaculation intensity,while serum BPA was positively correlated with premature ejaculation.According to the four percentile of BPA concentration (ng/ml),the subjects were divided into four groups.Compared with the low concentration group (0.38-3.79 ng/ml),the risk of sexual dysfunction significantly increased in the groups with higher BPA levels.Particularly,in the highest BPA group (8.68-21.93 ng/ml),more obvious effects were seen on sexual dysfunction (OR=1.55,95％CI:1.00-3.23),reduced sexual desire (OR=4.75,95％CI:2.44-9.22),reduced erection ability (OR=2.40,95％CI:1.18-4.88),reduced ejaculation intensity (OR=2.53,95％ CI:1.25-5.16) and premature ejaculation (OR=1.95,95％ CI:1.02-3.72).Conclusion Low sexual desire appeared as the main type of male sexual dysfunction,the exposure to higher levels of BPA in daily life might lead to male sexual dysfunction.
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Objective To explore the relationship between abnormal expression of fragile histidine triad (FHIT) gene and methyl-CpG-binding protein 2 (MeCP2) as well as their interaction on cervical cancerization.Methods A total of 73 patients with cervical squamous cell carcinoma (SCC),113 patients with cervical intraepithelial neoplasia (CIN Ⅰ,n =45;CIN Ⅱ/Ⅲ,n=68) and 60 women with normal cervix (NC) were included in the study.Real time PCR and Western blot were performed to detect the expression levels of mRNA and protein about FHIT and MeCP2,respectively.The methylation status of FHIT gene CpG island was tested by methylation-specifc PCR (MSP).Kruskal-Wallis H test,x2 test,trend x2 test and Spearman correlation analysis were conducted with software SPSS 20.0.The interaction was evaluated by generalized multifactor dimensionality reduction (GMDR) model.Results With the deterioration of cervical lesion,the methylation rates of FHIT gene CpG island (x2=18.64,P＜0.001;trendx2=18.08,P＜0.001) increased gradually,while the expression levels of FHIT mRNA (H=27.32,P＜0.001;trendx2=12.65,P＜0.001) and protein (H=47.10,P＜0.001;trendx2=29.79,P＜0.001) decreased gradually.There was a negative correlation between the methylation rates of FHIT gene CpG island and the expression level of FHIT protein (r=-0.226,P＜0.001).The levels of MeCP2 mRNA (H=26.19,P＜0.001;trend x2=11.81,P=0.001) and protein (H=69.02,P＜ 0.001;trend x2 =47.44,P＜ 0.001) increased gradually with the aggravation of cervical lesions.There was a positive correlation between the expression level of MeCP2 protein and the FHIT mRNA Ct ratio (r=0.254,P＜0.001).Expression of proteins were negatively correlated between MeCP2 and FHIT (r=-0.213,P=0.001).The results analyzed by GMDR model showed that there were interactions among high MeCP2 protein expression,the CpG island methylation of FHIT and mRNA and protein expression in CIN Ⅱ/Ⅲ group,and among high MeCP2 mRNA and protein expression,the CpG island methylation of FHIT and low mRNA and protein expression in SCC group.Conclusion High expression of MeCP2 mRNA and protein,the CpG island methylation and low mRNA and protein expression of FHIT could increase the risk of cervical carcinogenesis,and there might be a synergistic effect on cervical carcinogenesis.
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Objective To investigate the effect of heterogeneous nuclear ribonucleoprotein K (hnRNP K) and its interaction with human papillomavims 16 (HPV16) on cervical intraepithelial neoplasia (CIN).Methods The participants included 67 women with normal cervix (NC),69 women with CIN Ⅰ and 68 women with CIN Ⅱ/Ⅲ in a community cohort of pathologically diagnosed women established in Jiexiu of Shanxi province,from June 2014 to June 2015.A structured questionnaire was used to collect the demographic data of the subjects and the related factors of cervical lesions.Cervical exfoliated cells and cervical tissues from biopsy or surgery were selected.The infection status of HPV16 was detected by flow-through hybridization.The protein expression levels of hnRNP K were evaluated by Western blot.SPSS 23.0 software was used to collate and analyze the data.To study the differences in demographic characteristics,related factors,hnRNP K protein and HPV16 infection among NC,CIN Ⅰ and CIN Ⅱ / Ⅲ groups,X2 test,trend x2 test,and Kruskal-Wallis H test were conducted.Multiple comparisons of hnRNP K protein in three groups were completed by using the Bonferroni method.The OR and its 95％CI of hnRNP K,HPV16 and CIN were calculated by using the unconditional logistic regression models.Two-way interactions between hnRNP K protein and HPV16 infection on CIN were analyzed by using additive model and related indicators.Results HPV16 infection rates were 10.4％ in women with normal cervix,14.5％ in women with CIN Ⅰ and 41.2％ in women with CIN Ⅱ/Ⅲ,respectively.The differences among three groups were significant (P＜0.001).Moreover,the infection rates of HPV16 gradually increased with the increasing severity of CIN (trend x2=18.512,P＜0.001).The differences in protein expression of hnRNP K among three groups were significant (H=48.138,P＜0.001) and the expressionincreased with the development of cervical lesionss (trend x2=21.765,P＜0.001).Results from the interaction analysis indicated that there were additive effects between high expression of hnRNP K protein and HPV16 in CIN Ⅱ/Ⅲ group compared with normal group (API=0.639,95％CI:0.083-1.196).In contrast,no such additive effect was found in CIN Ⅰ group.Conclusions HPV16 infection and over-expression of hnRNP K protein were associated with the increased risk of cervical intraepithelial neoplasia.There might be interaction between hnRNP K protein overexpression and HPV16 infection existed on the progress of CIN Ⅰ/Ⅲ.
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Objective To explore the relationship between male sexual function and daily exposure to bisphenol A (BPA) at a reproductive center in Taiyuan.Methods Male patients who were seeking treatment of infertility due to problems caused by either of the spouse were selected from the Shanxi reproductive center between September 2014 and April 2015.Information on general characteristics,sexual function and fasting venous blood samples were collected.Total scores of sexual function were evaluated by Delphi expert scoring method.Levels of serum BPA were measured by high-performance liquid chromatography.Data was analyzed by Spearman rank correlation,rank sum test,multivariate logistic regression analysis andx2 trend test.Relationship between BPA and sexual function was presented as OR and corresponding 95％CI.Results Among the 353 participants,45.0％ was defined as sexual dysfunction with low sexual desire (47.3％) as the major reason.BPA was detected in all the 353 patients,with a range of concentration as 0.38-21.93 ng/ml and an average as 5.79 ng/ml.Results from the Spearman rank correlation analysis revealed significant negative correlations between serum BPA and sexual function,sexual desire,erectile ability and ejaculation intensity,while serum BPA was positively correlated with premature ejaculation.According to the four percentile of BPA concentration (ng/ml),the subjects were divided into four groups.Compared with the low concentration group (0.38-3.79 ng/ml),the risk of sexual dysfunction significantly increased in the groups with higher BPA levels.Particularly,in the highest BPA group (8.68-21.93 ng/ml),more obvious effects were seen on sexual dysfunction (OR=1.55,95％CI:1.00-3.23),reduced sexual desire (OR=4.75,95％CI:2.44-9.22),reduced erection ability (OR=2.40,95％CI:1.18-4.88),reduced ejaculation intensity (OR=2.53,95％ CI:1.25-5.16) and premature ejaculation (OR=1.95,95％ CI:1.02-3.72).Conclusion Low sexual desire appeared as the main type of male sexual dysfunction,the exposure to higher levels of BPA in daily life might lead to male sexual dysfunction.
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Objective To explore the relationship between abnormal expression of fragile histidine triad (FHIT) gene and methyl-CpG-binding protein 2 (MeCP2) as well as their interaction on cervical cancerization.Methods A total of 73 patients with cervical squamous cell carcinoma (SCC),113 patients with cervical intraepithelial neoplasia (CIN Ⅰ,n =45;CIN Ⅱ/Ⅲ,n=68) and 60 women with normal cervix (NC) were included in the study.Real time PCR and Western blot were performed to detect the expression levels of mRNA and protein about FHIT and MeCP2,respectively.The methylation status of FHIT gene CpG island was tested by methylation-specifc PCR (MSP).Kruskal-Wallis H test,x2 test,trend x2 test and Spearman correlation analysis were conducted with software SPSS 20.0.The interaction was evaluated by generalized multifactor dimensionality reduction (GMDR) model.Results With the deterioration of cervical lesion,the methylation rates of FHIT gene CpG island (x2=18.64,P＜0.001;trendx2=18.08,P＜0.001) increased gradually,while the expression levels of FHIT mRNA (H=27.32,P＜0.001;trendx2=12.65,P＜0.001) and protein (H=47.10,P＜0.001;trendx2=29.79,P＜0.001) decreased gradually.There was a negative correlation between the methylation rates of FHIT gene CpG island and the expression level of FHIT protein (r=-0.226,P＜0.001).The levels of MeCP2 mRNA (H=26.19,P＜0.001;trend x2=11.81,P=0.001) and protein (H=69.02,P＜ 0.001;trend x2 =47.44,P＜ 0.001) increased gradually with the aggravation of cervical lesions.There was a positive correlation between the expression level of MeCP2 protein and the FHIT mRNA Ct ratio (r=0.254,P＜0.001).Expression of proteins were negatively correlated between MeCP2 and FHIT (r=-0.213,P=0.001).The results analyzed by GMDR model showed that there were interactions among high MeCP2 protein expression,the CpG island methylation of FHIT and mRNA and protein expression in CIN Ⅱ/Ⅲ group,and among high MeCP2 mRNA and protein expression,the CpG island methylation of FHIT and low mRNA and protein expression in SCC group.Conclusion High expression of MeCP2 mRNA and protein,the CpG island methylation and low mRNA and protein expression of FHIT could increase the risk of cervical carcinogenesis,and there might be a synergistic effect on cervical carcinogenesis.
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Objective To explore the effect of Src on cervical cancer cells through ERK signal transduction pathway.Methods Experimental study was carried out in vitro.Cervical cancer cell lines Hela (HPV-positive) and C33A (HPV-negative) were treated with Src kinase inhibitor PP2.Then,the cell cycle and apoptosis of each group were evaluated by using flow cytometry (FCM).Western blotting and Real-time PCR were used to detect the levels of the expression of ERK 1/2,c-Fos and c-Jun mRNA and protein respectively.The database was established and analyzed with SPSS statistical software (version 20.0).Results After down-regulating Src,the cell proliferation was inhibited and cell apoptosis was induced.The proportions of G0/G 1 stage of Hela and C33A cell in cell cycle increased while G2/M and S stages decreased.Meanwhile,the mRNA levels of ERK 1,ERK 2,c-Fos and c-Jun increased.And the expression levels of ERK 1/2,phosphorylated ERK 1/2 (p-ERK 1/2)and phosphorylated c-Fos (p-c-Fos) protein decreased,while c-Jun and phosphorylated c-Jun (p-c-Jun)protein expression increased.In addtion,the change level of Hela cell,p-ERK 1/2 and c-Fos protein were lower than that of C33A cell before and after the Src inhibition.Conclusions Src,involved in regulating the expression of key factors of the ERK signal transduction pathway including p-ERK 1/2 and p-c-Fos,might be capable of promoting the proliferation of cervical cancer cells and inhibiting their apoptosis.The infection with HPV might have adjustable effect on this process.
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Objective To explore the effect ofpolycyclic aromatic hydrocarbons (PAHs) and p16,FHIT gene CpG island methylation,as well as their interaction in cervical intraepithelial neoplasias.Methods Objects of this study were from a cohort of cervical lesions study in Yangqu county of Shanxi province.All the patients were diagnosed pathologically,that including 83 patients with high-grade cervical intraepithelial neoplasia (CIN Ⅱ/Ⅲ),86 patients with low-grade cervical intraepithelial neoplasia (C1N Ⅰ) and another 91 women under normal cervical (NC) condition.1-hydroxy pyrene in the urine was detected by high performance liquid chromatography (HPLC)while CpG island methylation status of tumor suppressor gene p16 and FHIT were measured by methylation-specifc polymerase chain reaction (MSP).Data were analyzed with Kruskal-Wallis H test,chi-square test and trend of chi-square test.Logistic regression models were used to estimate the odds ratio (OR) and corresponding 95％ confidence intervals (95％CI) between influencing factors and the cervical disease by using the SPSS statistical software (version 20.0).The interaction under study was evaluated by using the generalized multifactor dimensionality reduction (GMDR) model.Results Level of 1-hydroxy pyrene (H=50.743,P＜0.001) and the high exposure rate of 1-hydroxy pyrene (trend x2=20.146,P＜0.001) were gradually increasing along with the severity of cervical intraepithelial neoplasia.The CpG island methylation rates ofpl6,FHIT in CIN] and CIN Ⅱ/Ⅲl group were higher than that in NC group,and gradually increasing along with the severity of cervical intraepithelial neoplasia (trend x2=9.75,P=0.002;trend x 2 =10.39,P=0.001).Results from the GMDR model showed that interaction existed among the high exposure of l-hydroxy pyrene and the CpG island methylation ofpl6,FHIT in CIN Ⅰ and CIN Ⅱ]/Ⅲ group.Conclusion Under the high exposure of 1-hydroxy pyrene and the CpG island methylation of p16,FHIT appeared to have increased the risk of cervical intraepithelial neoplasia and causing synergistic effect in cervical intraepithelial neoplasia.
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Objective To explore the effect of Src on cervical cancer cells through ERK signal transduction pathway.Methods Experimental study was carried out in vitro.Cervical cancer cell lines Hela (HPV-positive) and C33A (HPV-negative) were treated with Src kinase inhibitor PP2.Then,the cell cycle and apoptosis of each group were evaluated by using flow cytometry (FCM).Western blotting and Real-time PCR were used to detect the levels of the expression of ERK 1/2,c-Fos and c-Jun mRNA and protein respectively.The database was established and analyzed with SPSS statistical software (version 20.0).Results After down-regulating Src,the cell proliferation was inhibited and cell apoptosis was induced.The proportions of G0/G 1 stage of Hela and C33A cell in cell cycle increased while G2/M and S stages decreased.Meanwhile,the mRNA levels of ERK 1,ERK 2,c-Fos and c-Jun increased.And the expression levels of ERK 1/2,phosphorylated ERK 1/2 (p-ERK 1/2)and phosphorylated c-Fos (p-c-Fos) protein decreased,while c-Jun and phosphorylated c-Jun (p-c-Jun)protein expression increased.In addtion,the change level of Hela cell,p-ERK 1/2 and c-Fos protein were lower than that of C33A cell before and after the Src inhibition.Conclusions Src,involved in regulating the expression of key factors of the ERK signal transduction pathway including p-ERK 1/2 and p-c-Fos,might be capable of promoting the proliferation of cervical cancer cells and inhibiting their apoptosis.The infection with HPV might have adjustable effect on this process.
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Objective To explore the effect ofpolycyclic aromatic hydrocarbons (PAHs) and p16,FHIT gene CpG island methylation,as well as their interaction in cervical intraepithelial neoplasias.Methods Objects of this study were from a cohort of cervical lesions study in Yangqu county of Shanxi province.All the patients were diagnosed pathologically,that including 83 patients with high-grade cervical intraepithelial neoplasia (CIN Ⅱ/Ⅲ),86 patients with low-grade cervical intraepithelial neoplasia (C1N Ⅰ) and another 91 women under normal cervical (NC) condition.1-hydroxy pyrene in the urine was detected by high performance liquid chromatography (HPLC)while CpG island methylation status of tumor suppressor gene p16 and FHIT were measured by methylation-specifc polymerase chain reaction (MSP).Data were analyzed with Kruskal-Wallis H test,chi-square test and trend of chi-square test.Logistic regression models were used to estimate the odds ratio (OR) and corresponding 95％ confidence intervals (95％CI) between influencing factors and the cervical disease by using the SPSS statistical software (version 20.0).The interaction under study was evaluated by using the generalized multifactor dimensionality reduction (GMDR) model.Results Level of 1-hydroxy pyrene (H=50.743,P＜0.001) and the high exposure rate of 1-hydroxy pyrene (trend x2=20.146,P＜0.001) were gradually increasing along with the severity of cervical intraepithelial neoplasia.The CpG island methylation rates ofpl6,FHIT in CIN] and CIN Ⅱ/Ⅲl group were higher than that in NC group,and gradually increasing along with the severity of cervical intraepithelial neoplasia (trend x2=9.75,P=0.002;trend x 2 =10.39,P=0.001).Results from the GMDR model showed that interaction existed among the high exposure of l-hydroxy pyrene and the CpG island methylation ofpl6,FHIT in CIN Ⅰ and CIN Ⅱ]/Ⅲ group.Conclusion Under the high exposure of 1-hydroxy pyrene and the CpG island methylation of p16,FHIT appeared to have increased the risk of cervical intraepithelial neoplasia and causing synergistic effect in cervical intraepithelial neoplasia.