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 evaluate the effect of preoperative chemotherapy on bulky cervical cancer by internal iliac arterial infusion. Methods One hundred and eighty-six patients with bulky cervical cancer were randomly divided into two groups: chemotherapy + radiotherapy group (C+R group, n=105) and radiotherapy group (R group, n=81). Patients in C + R group underwent internal iliac arterial infusion chemotherapy by using Seldinger technique internal iliac arterial or epigastric arterial catheterization. Combined regimens were prescribed including cisplatin as the major drug. Meanwhile ~192 Ir high-dose-rate intracavitary radiotherapy was performed,with A point dose at ~12-24 Gy/2-4 times (C + R group). Patients in R group were only given radiotherapy. Both groups of patients received radical hysterectomy two weeks after radiotherapy. Results The tumor regression rate of C+R group was 97.1%, significantly higher than 79.0% in R group(P0.05). Postoperative pathologic examinations showed the percentage of cervical tumor residue, parauterine invasion,pelvic lymph node metastasis in C+R group was lower than those of R group(P

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.

Objecfive To investigate the relationship between single nucleotide polymorphism-56(SNP-56)in calpain-10(CAPN-10)gene and polycystic ovary syndrome(PCOS)in Chinese.Methods The genotypes of SNP-56 of CAPN-10 were determined through polymerase chain reaction Tm-shift genotyping method in 638 local women in Shandong Province.Among them,334 were patients with PCOS (PCOS group)and 304 were normal women(control group).The baseline parameters including levels of serum follicle-stimulating hormone(FSH),luteinizing hormone(LH),prolactin(PRL),estradiol(E2),testosterone(T)and lipid,as well as the body mass index(BMI)and waist/hip ratio(WHR)were measured.Glucose tolerance and insulin releasing before and after loading with 75 g of glucose were also assayed.Results(1)The frequencies of two allelotypes or three genotypes did not differ between PCOS women and normal women(P＞0.05).(2)In PCOS group,patients with AA genotype had a significantly higher plasma glucose of 180 minutes OGTT(5.7±2.2)mmol/L[P＜0.01 compared to Gagenotype(4.9±1.2) mmol/L,P＜0.01 compared to GG genotype(4.9±1.4)mmol/L]and serum total cholesterol(TC)level(4.9±1.0)mmol/L[P＜0.05 compared to Gagenotype(4.5±0.9)mmol/L].(3)Compared to PCOS patients with GA+GG genotype(P＜0.05,P＜0.01)or GG genotype(P＜0.05,P＜0.01),there was significantly higher attack rate of diabetes and tumor in the family history of patients with AA genotype.Conclusions These findings suggest that CAPN-10 gene SNP-56 which may not contribute to the genetic susceptibility of PCOS plays a role in glucose and lipid metabolism in Chinese PCOS patients.It may also be correlated with attack rate of diabetes and tumor in the family history of PCOS patients.

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 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 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.

Abstract: In order to investigate the feasibility of the extract from Cyclocarya paliurus as an additive in cigarettes, the volatile aromatic components were analyzed by gas chromatography-ion mobility spectrometry (GC-IMS), and C.paliurus extract was pyrolyzed to simulate cigarette smoking by TGA-GC/MS. The cracking products of C.paliurus were analyzed in a nitrogen environment, and the possible cracking mechanism of the products was reasonably speculated. The results showed that aldehydes, alcohols, and ketones were the primary volatile aroma components of the C.paliurus extract, comprising 62.28% of the total aroma components. The cracking products of C.paliurus extract varied greatly under different temperature conditions. A total of 79 compounds were identified at 200,360, and 440 ℃, among which 24 aromatic components were clearly identified as having significant effects on cigarette style, including aldehydes, ketones, alcohols, phenols, furans, benzene series, and other natural aromatic substances. Among them, phenols containing a high concentration are mainly formed by compounds containing the structure of guaiacol unit and eugenol unit by side chain cleavage, demethylation, demethoxylation, dehydration, etc. Furan and its derivatives are mainly formed by glycosides or glycoside compounds by breaking glucoside bonds and dehydration.

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.