Objective To investigate the expressions of human papillomavirus (HPV) 16E6 and E-cadherin in different cervical tissues.Methods Immunohistochemistry expressions of HPV16E6 and E-cadherin were analyzed in 30 normal cervical tissues,34 cervical intraepithelial neoplasias (CIN),and 60 squamous cervical cell carcinomas.Results The differences in positive expression rates of HPV16E6 and E-cadherin were statistically significant among normal cervical tissues,CIN Ⅰ,CIN Ⅱ-Ⅲ,cervical squamous cell carcinomas (x2 =40.166,P =0.000;x2 =31.295,P =0.000),respectively.The abnormal expressions of HPV16E6 and E-cadherin expression were significantly related to clinical pathological stage,tumor size,histological grade,invasion depth of squamous cervical cancers (P ＜ 0.05),but not correlated with age (P ＞0.05).The expressions between HPV16E6 and E-cadherin protein were negatively correlated (rs =-0.647,P =0.000).Conclusions There may be relationship between HPV16E6 and E-cadherin,and the abnormal expressions of two proteins may be associated with poor prognosis of cervical carcinomas.

Objective To investigate the influence of human papillomavirus (HPV) 16 E6 gene silencing by small interfering RNA (siRNA) on the expression and the promoter hypermethylation status of E-cadherin (E-cad) in cervical cancer SiHa cell line. Methods siRNA which used lentivirus as the vector was used to knock down the HPV16E6 gene in cervical cancer SiHa cell line. The expression levels of HPV16E6 mRNA, E-cad mRNA and protein in siRNA-HPV16E6 SiHa cell line were detected by RT-qPCR and Western blot, respectively. Methylation specific PCR (MSP) method was used to detect the methylation status of E-cad gene (CDH1) promoter in siRNA-HPV16E6 SiHa cell line. Results The E-cad mRNA expression levels in siRNA E6 group, empty vector group and blank control group were 4.755±1.085, 1.224± 0.840, 1.327±1.221, respectively. The protein expression levels were 0.616±0.019, 0.325±0.016, 0.299±0.015, respectively. The expressions of E-cad mRNA and protein in siRNA E6 group were significantly higher than those in the empty vector group and blank control group (F = 21.346, P < 0.01; F = 323.398, P < 0.01), and the difference between the empty vector group and blank control group was not statistically significant (P >0.05). After knocking down HPV16E6 gene, the methylation status of E-cad gene was weakly positive, and the intensity of the amplified products was significantly weaker than that in the empty vector group and blank control group, while the unmethylation amplification was positive. Conclusions Knocking down the HPV16E6 gene increases the expression of E-cad in cervical cancer SiHa cell line, and decreases the level of CDH1 promoter methylation. To a certain extent, it partly reverses the hypermethylation status of CDH1 promoter, and causes E-cad to be re-expressed.

Objective To investigate the value of DNA ploidy analysis in cervical cancer screening for outpatients.Methods 840 from 2 692 outpatients examed by Thin Prep cytology,DNA ploidy analysis were performed directed biopsy simultaneously.Sample were taken by cervix brush and transported into a fixative solution.Two slides were made from each sample for staining with Feulgen DNA specific staining and the other with Pap stained,respectively.The routine cytological diagnosis of Pap smear was done by cytology physicians,and the Feulgen staining tablets by the automated DNA ploidy analysis system.Results Among 840 cases,554 cases (66.0 ％) were histological diagnosed as chronic cervicitis,25 cases (3.0 ％) as cervical intraepithelial neoplasia (CIN) Ⅰ,59 cases (7.0 ％) as CIN Ⅱ,100 cases (11.9 ％) as CINⅢ and 102 cases (12.1％) as cervical invasive cancer by pathological biopsy.486 cases were observed with DNA heteroploid and 354 were not.The sensitivity,specificity,positive predictive values and negative predictive values of scanning CIN Ⅱ or more severe cervical diseases by DNA heteroploid positive or heteroploid ≥3 for were 91.9 ％ or 89.2 ％,58.5 ％ or 35.8 ％,49.4 ％ or 57.3 ％,94.1％ or 77.2 ％,respectively,while those of scanning equal or more than LSIS andthe above diseases by Thin Prep cytology were 40.2 ％,90.0 ％,39.6 ％ and 76.9 ％.Conclusion DNA ploidy analysis might be a useful tool for cervical cancer screening and has a competitive sensitivity compared with conventional cytology.

Objective To investigate the effects of omega-3 polyunsaturated fatty acids (ω-3 PUFA)supplementation on brain edema,autophagy response and neurobehavioral outcome after traumatic brain injury (TBI) in rats and the related mechanisms.Methods TBI rat models were established using Feeney's method.Seventy-two SD rats were divided into 4 groups using random number table:sham operation group,TBI group,ω-3 PUFA supplementation group (TBI + ω-3 group) and autophagy inhibitor 3-methyladenine group (TBI + 3-MA group) (all n =18),each group was further divided into 3 sub-groups (n =6) corresponding to 3 time points (days 1,3,and 7 after TBI).On each of the 3 time points,we measured rat behavioral outcomes with modified neurologic severity score (mNSS) tests;brain water content was measured with wet-dry weight method.The mRNA and protein expressions of autophagy-related factors (LC3-Ⅱ and Beclin-1) in TBI cerebral cortex were determined by immunohistochemistry staining,reverse transcription-polymerase chain reaction and Western blot on day 3 after TBI.Results Compared with the sham group,on days 1,3,and 7 after injuary,the TBI group,the TBI + ω-3 group,and the TBI + 3-MA group had significantly higher mNSS scores (TBI group:12.42±0.27vs.1.34±0.32,12.07±0.27vs.1.16±0.29,10.22±0.39vs.1.22±0.30;TBI+ω-3 group:12.05 ±0.23 vs.1.34 ±0.32,11.38 ±0.21 vs.1.16±0.29,8.20 ±0.21 vs.1.22±0.30;TBI +3-MA group:11.93 ±0.20 vs.1.34 ±0.32,11.09 ±0.19 vs.1.16 ±0.29,7.93 ±0.17 vs.1.22 ± 0.30;all P =0.00) and brain water content [TBI group:(79.82 ± 0.61) ％ vs.(71.87 ± 0.43) ％,(83.04±0.42)％ vs.(72.13 ±0.53)％,(75.12 ±0.72)％ vs.(71.78 ±0.38)％;TBI+ω-3 group:(76.81 ±0.63)％ vs.(71.87 ±0.43)％,(79.39 ±0.59)％ vs.(72.13 ±0.53)％,(73.86 ±0.38)％ vs.(71.78 ±0.38)％;TBI+3-MAgroup:(75.98 ±0.49)％ vs.(71.87 ±0.43)％,(77.14 ±0.46)％ vs.(72.13 ±0.53)％,(72.24 ±0.37)％ vs.(71.78 ±0.38)％;all P =0.00].The mRNA and protein expressions of LC3-Ⅱ and Beclin-1 in the brain were also significantly higher on day 3 in the TBI group,the TBI + ω-3 group,and the TBI + 3-MA group (all P =0.00).Compared with the TB1 group,on day 3 and day 7 after injury,the TBI + ω-3 group and the TBI + 3-MA group had significantly lower mNSS scores (TBI + ω-3 group:11.38±0.21 vs.12.07±0.27,P=0.04,8.20±0.21 vs.10.22±0.39,P=0.01;TBI+3-MA group:11.09±0.19vs.12.07 ± 0.27,P=0.01,7.93 ± 0.17 vs.10.22±0.39,P=0.00).Ondays1,3,and 7,compared with the TBI group,the TBI + ω-3 group and the TBI + 3-MA group had significantly lower brain water content [TBI + ω-3 group:(76.81 ± 0.63) ％ vs.(79.82 ± 0.61) ％,P =0.04,(79.39 ±0.59)％ vs.(83.04±0.42)％,P=0.01,(73.86±0.38)％ vs.(75.12±0.72)％,P=0.03;TBI+3-MAgroup:(75.98 ±0.49)％ vs.(79.82 ±0.61)％,P=0.01,(77.14 ±0.46)％ vs.(83.04 ±0.42)％,P =0.00,(72.24 ± 0.37) ％ vs.(75.12 ± 0.72) ％,P =0.02].On day 3,the TBI + ω-3 group and the TBI + 3-MA group had significantly reduced LC3-Ⅱ and Beclin-1 mRNA expression compared with the TBI group (TBI +ω-3 group:P=0.04,P =0.01;TBI +3-MA group:P =0.01,P =0.00) and protein expression (TBI+ω-3 group:P=0.01,P=0.03;TBI +3-MA group:both P=0.00).Conclusion ω-3 PUFA supplementation could markedly reduce brain edema and improve neurological functions after TBI,showing a neuroprotective effect,possibly through inhibiting TBI-induced autophagy responses.

Objective To measure the level of interleukin (IL)-27 in sera of patients with Sj(o)gren's syndrome (SS) and investigate its role in IL-10 in CD3+CD8-T cells.Methods Serum levels of 1L-27 and IL-10 from 34 SS patients and 33 healthy controls were tested by enzyme linked immunosorbent assay (ELISA).Peripheral blood monocytes (PBMCs) from SS patients were collected and cultured in different conditions (one with PHA,the other with PHA and IL-27).Seventy-two hours later,cells were harvested and the percentages of CD3 +CD8-IL-10+ cells were measured by flow cytometry.Expressions of c-Maf,IL-10 transcription factor,were examined using real-time polymerase chain reaction (PCR).The data were analyzed by t test,Pearson's correlation analysis and multiple linear regression analysis.Results Serum level of IL-27 in patients with SS [(158±34) pg/ml] was significantly higher than that in healthy controls [(139±18) pg/ml,t=2.823,P＜0.01].IL-27 level was positively correlated with IL-10 and IgG levels (r=0.384,P＜0.05; r=0.354,P＜0.05),but negatively correlated with SSDAI (r=0.503,P＜0.01).Multiple regression analysis showed independent correlation of IL-27 with IL-10 and SSDAI in SS patients (β=0.412,P＜0.01; β=-0.525,P＜0.01),but not IgG.CD3+CD8-IL-10+ cells was upregulated by IL-27 in vitro (P＜0.01,n=14),so was c-Maf (P＜0.01,n=22).Conclusion IL-27 could ameliorate disease activity of Sjogren's syndrome.

Objective To explore the security,pregnancy outcomes,and the tumor recurrence related factors of young patients with cervical cancer treated with different radical trachelectomy (RT).Methods Thirty-two young patients ＜ 40 years of age with early cervical cancer from May 2004 to July 2012 admitted in Tumor Hospital Xiangya School of Medicine of Central South University were divided into two groups based on different operation methods:vaginal radical trachelectomy (RVT) group and abdominal radical trachelectomy (RAT) group.The clinical data were analyzed by One-way Anova and multivariate Cox stepwise regression analysis.Results The operation duration,number of lymph node dissection,the height of the cervical resection,postoperative hospitalization time,incidence of vascular injury and incidence of postoperative lymphocele were respectively (250 ± 82) min,15 ± 6,(2.31 ± 0.21) cm,(9.2 ± 2.9) d,1/18 and 1/18 in RVT group,while (263 ±60) min,16 ±8,(2.32 ±0.26) cm,(10.3 ±3.5) d,0 and 1/14 in RAT group.There was no statistically significant difference between the two groups (all P ＞ 0.05).The blood loss (281 ±201) ml in RVT group was significantly lower than that in the RAT group (492 ±320) ml (P ＜0.05).The length of Vaginal hysterectomy[(2.61 ±0.50) cm] and the width of parametrial resection[(2.38 ±0.36) cm] in RVT group were significantly less than those[(2.95 ±0.10),(2.81 ±0.22) cm] in the RAT group (all P ＜ 0.05).The pregnancy rate between RVT group (3/18) and RAT group (2/14) were no significant difference (P ＞ 0.05).One-way Anova analysis showed that the recurrence of early cervical cancer was related to tumor size in diameter (F =4.911,P =0.047),while there were no correlation with age,clinical stage,histological type and surgical approach (all P ＞ 0.05).Multivariate analysis showed that tumor diameter size was an independent risk factor for tumor recurrence (3 =0.259,P =0.031).Conclusions RT for young patients with early cervical cancer is feasible.Pregnancy outcomes after RT need to be study in the future.Tumor size in diameter is the major risk factor for tumor recurrence.

Total hysterectomy is a treatment for cervical intraepithelial neoplasia Ⅲ (CINⅢ), and its recurrence rate is very small. In recent years, with the development of colposcopy and the increase of postoperative follow-up rate, the detection rate of vaginal intraepithelial neoplasia (VAIN) and vaginal cancer after CINⅢ hysterectomy has increased significantly. In recent years, a number of studies have found that persistent human papillomavirus (HPV) infection is the most important factor leading to the recurrence of vaginal stump after CINⅢ level hysterectomy, so eliminating HPV infection can improve the patient's performance. The prognosis of persistent HPV infection is affected by many factors. The author reviewed the factors related to the clearance of HPV after CINⅢ hysterectomy. The purpose of this study is to provide a theoretical basis for improving the prognosis of patients and delaying the recurrence time after CINⅢ hysterectomy.

Objective To investigate the effects of specific methyltransferase inhibitor of 5-Aza-2'-deoxycytidine (5-Aza-CdR) on the promoter methylation of E-cadherin (E-cad) gene, protein expression in human cervical cancer SiHa cells, and the cell biological behavior. Methods SiHa cells were treated with 5-Aza-CdR at different concentrations. Quantitative methylation-specific polymerase chain reaction (QMSP) was used to examine CpG island promoter methylation level of E-cad gene before and after treatment. The experimental group of the optimum concentration was selected. The expression levels of E-cad mRNA and its protein in SiHa cells line were detected by quantitative real-time polymerase chain reaction (RT-PCR) and western blot respectively. Cell adhesion test was used to measure cell adhesion ability and Transwell test was used to detect cell invasion and migration ability. Results E-cad gene promoter methylation index (PMR) of 5-Aza-CdR at 0, 1, 5, 10, 15 μmol/L level was (53.0 ±1.6) %, (50.0 ±1.2) %, (44.0 ±1.4) %, (27.0 ±1.7) %, (15.0±8.2) %respectively, and PMR value decreased gradually with the increase of 5-Aza-CdR concentration. Furthermore, PMR value was the lowest at 15μmol/L, and the difference was statistically significant compared with other 4 groups (P< 0.01). Then 5-Aza-CdR at 15 μmol/L was selected as the following experimental concentration. The expression of E-cad mRNA and its protein in the 5-Aza-CdR group were significantly higher than those in the blank control group (P<0.05). The rates of cell adhesion , cell invasion inhibition and migration inhibition were all increased with significant differences (P<0.05). Conclusions 5-Aza-CdR can upregulate E-cad mRNA and protein expression level in cervical cancer SiHa cells, reduce cell invasion and migration ability, and promote the adhesion of SiHa cells, which has reversed hypermethylation in the promoter region of E-cad gene partly.

Malignant ovariangerm cell tumors (MOGCTs) is second only to epithelial tumor which often occur in young women and young women,with high malignancy and high mortality.Effective treatment is particularly important in clinical practice.The prognosis is improved for valid chemotherapy scheme foun ded in recent years.Surgery still play a crucial role in the therapy of MOGCTs no matter for the primary operation or re-operation.Since the 70s,the comprehensive surgical staging (CSS) has improved the prognosis in patients with malignant ovarian cell tumors.Retroperitoneal lymphadenectomy is an integral part of the complete staging in MOGCT.The paper discusses various aspects of the clinical value of comprehensive surgical staging in MOGCTs.

Acupuncture therapy needs to cause enough somatosensory stimulation to achieve therapeutic effect.How to identify the site and depth before acupuncture,assess therapeutic effect after acupuncture and ensure the safety of treatment are the points for clinical application of acupuncture therapy,but no standard objective and quantitative access has been achieved.Under assistance of ultrasound,precise positioning before acupuncture,visualization of anatomical structures and quantification of the depth during acupuncture became feasible.The research progresses of ultrasound for assisting acupuncture therapy were reviewed in this article.