Objective To confirm the role of HLA B2704 in the pathogenesis of psoriasiform dermatosis by investigating the phenotype of skin diseases in HLA B2704 transgenic mice.Methods The transgenic positive mice were screened and identified by PCR,dot blot and Southern blot hybridization.HE and Immunohistochemistry staining of the diseased mice skin were performed to detect the pathological changes and expression of HLA B2704.Results Six transgenic positive mice developed obvious psoriasiform dermatosis.Histologically,the skin was massively thickened by psoriasiform hyperplasia and keratinization.HLA B2704 antigen was highly expressed on the cell surface.Conclusion HLA B2704 heavy chain can induce the psoriasiform dermatosis in the transgenic mice.

AIM: To study the effect and mechanism of chlorophyllin (CHL) inhibiting HT29 cells. METHODS: IC 50 value and growth curve of HT29 cells were detected with MTT method. Apoptosis was detected with Wright-Giemsa staining, FCM and DNA electrophoresis. Telomerase was detected by PCR-ELISA, and protein and mRNA expression of COX-2 gene were detected through RT-PCR and Western blot. RESULTS: CHL inhibited the growth of HT29 in a dose-dependent manner. CHL blocked HT29 cells in G 1 phase but did not induce apoptosis. Different concentration of CHL inhibits the expression of telomerase and COX-2 in HT29 cells. CONCLUSION: CHL inhibits the growth of HT29 cells by inhibiting the expression of telomerase and COX-2 and blocking cells in G 1 phase. [

Objective: To explore the effect of vaginal micro-environment alterations and HPV16 infection and their interaction in the progression of cervical intraepithelial neoplasia. Methods: The participants of this study came from the cervical lesions study cohort in Shanxi province, including 623 women with normal cervical (NC), 303 patients with pathogenically diagnosed low-grade cervical intraepithelial neoplasia (CINⅠ) and 93 patients with pathogenically diagnosed high-grade cervical intraepithelial neoplasia (CINⅡ/Ⅲ). The data of the demographic characteristics of the study subjects and factors related to cervical intraepithelial neoplasia were collected, and HPV16 infection were detected by using flow-through hybridization technology and H(2)O(2), β-glucuronidase, clotting enzyme, neuraminidase and leucocyte esterase in vaginal secretions were detected by using the combined detection kit of aerobic vaginitis and bacterial vaginosis. pH value and vaginal cleanliness were also detected at the same time. The database was established and analyzed by SPSS statistical software (version 22.0). Results: The HPV16 infection rate (trend χ(2)=55.45, P<0.001) and the abnormal rates of H(2)O(2) (trend χ(2)=26.19, P<0.001), pH (trend χ(2)=5.06, P=0.024), vaginal cleanliness (trend χ(2)=19.55, P<0.001), β-glucuronidase (trend χ(2)=17.52, P<0.001) and neuraminidase (trend χ(2)=14.90, P<0.001) increased gradually along with the severity of cervical intraepithelial neoplasia, but the abnormal rates of clotting enzyme and leucocyte esterase showed no same trend. The results of GMDR model analysis showed that there was interaction between HPV16 infection and abnormalities of H(2)O(2), β-glucuronidase, clotting enzyme and neuraminidase in CINⅠ group, and the interaction between HPV16 infection and the abnormalities of vaginal cleanliness, H(2)O(2), β-glucuronidase and neuraminidase in CIN Ⅱ/Ⅲ group. Conclusion: Our findings indicated that the vaginal micro-environment alterations and HPV16 infection could increase the risk of cervical intraepithelial neoplasia, and they might have an important synergistic effect in the progression of cervical intraepithelial neoplasia.
Female ; Human papillomavirus 16/isolation & purification* ; Humans ; Hydrogen Peroxide ; Papillomavirus Infections/virology* ; Uterine Cervical Neoplasms/virology* ; Uterine Cervical Dysplasia/virology*

Objective: To investigate the effect of heterogeneous nuclear ribonucleoprotein K (hnRNP K) and its interaction with human papillomavirus 16 (HPV16) on cervical intraepithelial neoplasia (CIN). Methods: The participants included 67 women with normal cervix (NC), 69 women with CINⅠ and 68 women with CINⅡ/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, CINⅠand CINⅡ/Ⅲgroups, χ(2) test, trend χ(2) 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 CINⅠ and 41.2% in women with CINⅡ/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 χ(2)=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 χ(2)=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 CINⅡ/Ⅲ group compared with normal group (API=0.639, 95%CI: 0.083-1.196). In contrast, no such additive effect was found in CINⅠ 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 CINⅡ/Ⅲ.
Case-Control Studies ; Disease Progression ; Female ; Gene Expression Regulation, Neoplastic ; Heterogeneous-Nuclear Ribonucleoprotein K/metabolism* ; Human papillomavirus 16 ; Humans ; Papillomavirus Infections ; Uterine Cervical Neoplasms/virology* ; Uterine Cervical Dysplasia/virology*

OBJECTIVE To establish and optimize the detection of highly pathogenic influenza virus H5N1 subtype by RT-Multiplex (RT-M) PCR method. METHODS The viral RNA was reversely transcripted with universal primer,and the cDNA was sequenced and analyzed. According to the cleavage site of H5 and the conservative sequence of N1,designed two pairs of specificity primer,RT-M PCR was developed by these primers. Then verified the sensitivity of method and its specificity by detecting comparing with NDV,IBV and IBDV. RESULTS The results of H5N1 gene sequence showed the high homology with the Anhui H5N1 strain [avian influenza A/Anhui/1/2005 (H5N1)] after comparing in the GenBank. Two fragments of 302 bp and 567 bp were amplified by RT-M PCR. The method could detect 0.500 pg virus RNA at least,and own high specificity. CONCLUSIONS This method can be used for highly pathogenic avian influenza.

AIM: To explore the association of single nucleotide polymorphisms of promoter G~ -597 -A, G~ -572 -C, G~ -174 -C and T~ 15 -A, C~ 132 -T in exon 5 of interleukin 6 and lumbar intervebral disc disease. METHODS: The single nucleotide polymorphisms of interleukin 6 gene, including polymorphisms of G~ -597 -A, G~ -572 -C in promoter, G~ -174 -C and T~ 15 -A, C~ 132 -T in exon 5 were analyzed by the polymerase chain reaction and sequencing methods in 81 cases with lumbar intervebral disc disease and 101 healthy controls. The association of single nucleotide polymorphisms of interleukin 6 gene with lumbar intervebral disc disease in two groups was measured. The association of single nucleotide polymorphisms of interleukin 6 gene with lumbar intervebral disc degeneration in those younger than 45-year-old were also measured. RESULTS: The G~ -572 -C polymorphism of interleukin 6 gene was observed, but no single nucleotide polymorphism of G~ -597 -A, G~ -174 -C in promoter and T~ 15 -A, C~ 132 -T in exon 5 in two groups was detected. There was no difference between the distribution of the G~ -572 -C polymorphism of interleukin 6 gene in two groups. In those younger than 45-year-old the association of the single nucleotide polymorphism of interlukin-6 gene and lumbar intervebral disc degeneration was not significant. CONCLUSION: There is G~ -572 -C polymorphism in Chinese. No relation between G~ -572 -C polymorphism of interleukin 6 with lumbar intervebral disc disease and lumbar intervebral disc degeneration was observed.

Objective:To improve the primary culture method of rat pulmonary microvascular endothelial cells(PMVECs) and obtain purified PMVECs.Methods:The modified tissue block pasted culture method was used to isolate and culture Wistar rat PMVECs. The morphous of cultured cells were observed by microscopy. The cultured cells were identified by detecting factor Ⅷ related antigen and binding isolectin B4. Results and Conclusion:The morphous of cultured primary PMVECs in vitro showed short fusiform shape or polygon, and the monolayer of cultured cells displayed the shape of pavingstone. But the morphous changed followed the transfer of culture and the change of culture condition. The cultured cells had characterization of binding isolectin B4 and negative immunocytochemical staining for factor Ⅷ related antigen. The cultured PMVECs have good growth state and purity,and can be subcultringed stably.The observation of cell morphous integrating with immunocytochemical staining is a reasonable identification method for PMVECs.

Objective To establish a method of isolation, purification, primary culture and identification of alveolar type Ⅱ epithelial cells(AT-Ⅱ).Methods The AT-Ⅱs were isolated from Wistar rats by trypsin,purified by differential centrifugation, erythrocyte spallation, differential adherence and immune adherence, and identified by observing the morphology of cultured cells under the inverted phase and tannic acid staining. Results and Conclusion The cultured primary AT-Ⅱs in vitro presented single or island form growth, and their shapes were round or elliptical. A great deal of fine particles showed sharp contrast, and were observed in intracytoplasm. The cell nuclei were clear. They were positive for tannic acid staining.The primary culture AT-Ⅱs obtained from improved isolation and purification have good growth state and purity, and are suitable for research in vitro.

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; CINⅡ/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, χ(2) test, trend χ(2) 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 (χ(2)=18.64, P<0.001; trend χ(2)=18.08, P<0.001) increased gradually, while the expression levels of FHIT mRNA (H=27.32, P<0.001; trend χ(2)=12.65, P<0.001) and protein (H=47.10, P<0.001; trend χ(2)=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 χ(2)=11.81, P=0.001) and protein (H=69.02, P<0.001; trend χ(2)=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 CINⅡ/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.
Acid Anhydride Hydrolases/metabolism* ; Carcinoma, Squamous Cell/pathology* ; DNA Methylation ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Methyl-CpG-Binding Protein 2/metabolism* ; Neoplasm Proteins/metabolism* ; Polymerase Chain Reaction/methods* ; RNA, Messenger ; Uterine Cervical Neoplasms/pathology* ; Uterine Cervical Dysplasia/pathology*

Objective: To evaluate the effects of polycyclic aromatic hydrocarbons (PAHs) and high risk human papillomavirus (HR-HPV) infection and their interaction on the progression of cervical intraepithelial neoplasia. Methods: A total of 486 patients, including 208 women with normal cervix (NC), 154 patients with low-grade cervical intraepithelial neoplasm (CINⅠ), 124 patients with high-grade cervical intraepithelial neoplasm (CINⅡ/Ⅲ), were selected from the cervical lesions cohort from June to December, 2014. HR-HPV was detected by using flow-through hybridization technology and the urine concentration of 1-hydroxypyrene (1-OHP) was detected with high performance liquid chromatography. By using software SPSS 22.0, the χ(2) test, trend χ(2) test, Kruskal-Wallis H test, Nemenyi rank test and Spearman rank correlation analysis were performed. And the interaction effects were evaluated by additive model. Results: The HR-HPV infection rates in NC, CINⅠ and CINⅡ/Ⅲ groups were 27.9%, 37.0% and 58.9%, respectively. The urine concentrations of 1-OHP (μmol/molCr) were 0.07±0.09, 0.11±0.10 and 0.17±0.15, respectively. With increasing severity of the cervical lesions, the HR-HPV infection rate gradually increased (trend χ(2)=29.89, P<0.001) and the high exposure rate of PAHs gradually increased (trend χ(2)=27.94, P<0.001). HR-HPV infection was positively correlated with 1-OHP exposure (r=0.680, P<0.001). There was a positive additive interaction between HPV infection and PAHs exposure in CIN Ⅱ/Ⅲ group, but it was not found in CIN Ⅰ group. Conclusion: Both HR-HPV infection and high exposure of PAHs might increase the risk of cervical intraepithelial neoplasm, and might have a synergistic effect on the progression of high-grade cervical intraepithelial neoplasia.
Case-Control Studies ; Cohort Studies ; Disease Progression ; Female ; Humans ; Papillomaviridae/isolation & purification* ; Papillomavirus Infections/virology* ; Polycyclic Aromatic Hydrocarbons/pharmacology* ; Pyrenes/urine* ; Severity of Illness Index ; Uterine Cervical Dysplasia/virology* ; Uterine Cervical Neoplasms/virology*