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 ectopic pregnancy from embryo transfer (ET)of in-vitro fertilization (IVF) cycle and intracytoplasmic sperm iujection (ICSI) cycle and frozen-thawed (FET) cycle.Methods From Jan.2005 to Dec.2010,a total of 9037 IVF-ET or ICSI-ET cycles and 4034 FET cycles were performed in our reproductive medicine center,Affiliated First Hospital of Zhengzhou University.The incidence of ectopic pregnancy rate was studied in fresh cycles IVF-ET (5998) and ICSI-ET (3039) cycles,and natural FET (2198) and hormone replacement (E-P) FET (1836) cycles.ResultsOf 4034 FET cycles,1090 clinical pregnancies and 26 ectopic pregnancies were observed,the incidence of ectopic pregnancy was 2.38％ (26/1090).Of 9037 fresh cycles,3602 cycles were clinical pregnancy,and 133 cycles were ectopic pregnancy,and the incidence of ectopic pregnancy was 3.69％ (133/3602).The ectopic pregnancy rate in FET cycles was lower than in fresh cycles significantly (P ＜0.05).Of 3039 fresh ICSI-ET cycles,the incidence of ectopic pregnancy was 2.62％ ( 34/1298 ) in 1298 clinical pregnancies.Of 5998 IVF-ET cycles,2304 clinical pregnancies were observed,the incidence of ectopic pregnancy was 4.30％ (99/2304).Ectopic pregnancy rate in the fresh ICSI-ET cycles was lower than that of IVF-ET group significantly (P ＜ 0.01 ).The ectopic pregnancy rate in the natural FET cycles was 1.46％ (8/547),which was significantly lower than 3.31％ ( 18/543 ) in E-P group ( P ＜ 0.05 ).ConclusionsThe incidence of ectopic pregnancy of FET cycles was significantly lower than that of fresh embryo transfer cycles.The application of exogenous sex hormones in assisted reproductive cycles might increase occurrence of ectopic pregnancy.

Hepatocellular carcinoma(HCC)is one of the malignant tumors with the highest cause of death and increasing incidence worldwide.Accurate diagnosis in early stage is vital for the treatment of patients.Presently,routine screening strategies including ?-fetoprotein(AFP)and ultrasound every 6 months have been recommended for early detection in patients with liver cirrhosis to detect HCC at earlier stages.However,the sensitivity and specificity of AFP are far from satisfaction.With the development of recently techniques such as proteomics,it is possible for new HCC-specific markers being available in the near future.Golgi Protein 73(GP73)is most likely to be a promising serum marker.In a few available literatures,GP73 has sensitivity and specificity of 69% and 90%.And fucose-studded GP73 has sensitivity and specificity of 90% and 100%.Apart from GP73,lens culinaris agglutinin-reactive AFP,des-gamma carboxyprothrombin,?-L-fucosidase,glypican-3,hepatocyte growth factor,transforming growth factor-?1,vascular endothelial growth factor,and mucin 1 have been proposed as markers for HCC detection.Clinical trials are undergoing to estimate the value of the above markers,and may change the status in quo ofthe diagnosis of HCC.We have started a multi-centeral trial to investigate GP73 in HBV and HBV related HCC patients with positive preliminary results.

Ojective To investigate the clinical application value of low dose contrast agent in computerized tomography pulmonary angiography (CTPA) with spectral CT imaging.Methods Totally 52 patients suspected with pulmonary embolism underwent multi-slice spiral CT pulmonary angiography,and were randomly divided into a control group (n =27) and a research group (n=25).The research group used spectral CT with 35 ml of contrast medium and the control group used 64-slice CT with conventional 80 to 90 ml of contrast medium.The CT values of the pulmonary trunk,left pulmonary artery (LPA),right pulmonary artery (RPA),pulmonary vein and ascending aorta were measured.The contrast to noise ratio (CNR) of the pulmonary artery was also calculated.In addition,the image quality of CTPA was evaluated independently by two experienced radiologists.Results The CT values (HU) of the pulmonary trunk,left pulmonary artery(LPA),right pulmonary artery (RPA) in the research group were (432.2±63.4),(373.5±48.8),(381.4±53.6) and (62.5-±6.4),respectively,and significantly higher than those in the control group,showing statistical differences (P＜0.05).The evaluation results of the CTPA images by two radiologists showed that the image quality in the research group was better than that in the control group (P＜0.05).Conclusion The image quality of pulmonary angiography with spectral CT using low contrast medium dose can be improved compared with the conventional spiral CT.

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.

Objective To measure the levels of cross-reactive neutralizing antibodies responding to various subtypes of HIV-1 strains in people with HIV-1 infection in Chongqing and to analyze their character-istics.Methods Two hundred and thirty-six plasma samples were collected from patients with chronic HIV-1 infection in Chongqing city.The single-round-infection assay in TZM-bl cells were performed to screen the plasma samples with cross-reactive neutralizing antibodies against various subtypes of HIV-1 strains by using HIV-1 pseudovirus strains and to measure the 50%inhibitory dose ( ID50 ) .Results Eight-een out of 236 (7.63%) plasma samples were able to neutralize various B subtypes and/or C subtypes of HIV-1 pseudovirus strains, among which 15 plasma samples showed the great ability to neutralize pseudovir-us strains of different subtypes.Conclusion The cross-reactive neutralizing antibodies against various sub-types of HIV-1 strains existed in plasma samples collected from people with chronic HIV-1 infection in Chongqing city.

Objective To explore the effects of umbilical cord-derived mesenchymal stem cells (UCMSCs) on fibroblast-like synoviocytes(FLSs) from patients with rheumatoid arthritis(RA). Methods collagen-induced arthritis(CIA) models were developed on Wistar rats and 1 ×106 UCMSCs were given by intravenous injection from tail vein on the 17th day. On day 42, rats were sacrificed and synovial tissues were obtained to detect matrix metalloproteinase (MMP)-1, MMP-3 and MMP-13. Synovial tissues from patients with RA and osteoarthritis(OA) treated by knee arthroplasty were used to isolate FLSs. RNA of FLSs were extracted to compare MMP-1, MMP-3 and MMP-13 expression. FLSs and UCMSCs were cocultured through transwell for 72 hours. Then levels of MMPs were compared in the supernatants by Luminex. The MMPs expressed by FLSs and soluble factors expressed by MSCs were detected by real time-polymerase chain reaction(PCR). After adding antibodies to soluble factors, the MMPs expressions of RA FLSs were compared. Results MMP-1 (6.9±5.4, 1.3±1.4, P<0.05), MMP-3 (6.0±6.5, 1.4±1.0, P<0.05) and MMP-13 (21.8± 20.8, 1.5±1.6, P<0.05) expression were much higher in CIA rats compared with healthy controls. MMP-1 (1.3±1.4, 6.9±5.4,8.7±6.8, P<0.05), MMP-3(1.4±1.5, 6.0±6.5, 6.0±5.7, P<0.05) and MMP-13(3.0±3.2,
22±21, 22±26, P<0.05) expression were inhibited by UCMSCs in vivo. In vitro, MMP-1 (1.8±0.9, 0.9±0.7, t=2.44, P<0.05), MMP-3(2.6±1.7, 1.1±1.0, t=2.25, P<0.05) and MMP-13(2.4±2.3, 0.6±0.7, t=2.37, P<0.05) levels were higher in RA than OA FLSs. After coculture, MMP-13(1.3±1.2, 0.9±1.2, t=3.63, P<0.05) expressed by FLSs were down-regulated, however MMP-1 (1.5±1.4, 6.6±6.0, t=3.90, P<0.05) and MMP-3 (7±17, 22±35, t=2.86, P<0.05) were up-regulated when analyzed by paired t-test. Soluble factors such as I-DO, HGF and IL-10 were elevated. Anti-IL-10 antibody could decrease the function of UCMSCs by inhibiting MMP-13 expression in RA FLSs. Conclusion UCMSCs ameliorates RA by secreting soluble factor IL-10, which may inhibit MMPs expressed by FLSs.

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 investigate the impact of the CD4+CD25nt/hiCD127lo regulatory T cell subset frequency on the immune status and disease progression of Chinese HIV-1 infected individuals.Methods:83 untreated HIV-infected individuals and 312 healthy control individuals of four distinct age groups were enrolled in the research. The CD4+ T cell absolute counts, phenotypes and frequency determination of CD4+CD25nt/hiCD127lo Regulatory T cell subsets was performed on freshly obtained whole blood samples by 3-color immune staining flow cytometry. The HIV-1 specific cellular immune function was test at single cell level by ELISpot. The corresponding plasma viral load was determined by NASBA.Results:The frequency of peripheral CD4+CD25nt/hiCD127lo regulatory T cells of HIV infected individuals in distinct disease progression status was dissimilar in China , and significantly increased in contrast to the healthy controls(P

Objective:To explore the association between left-behind status, screen time (ST) and behavior of autism in rural preschool children.Methods:Cross section study was used in this study.A sample of 3 636 rural preschool children aged from 3 to 6 years old in 26 kindergartens were selected from four counties in Anhui province of China.The contents of the questionnaire include: basic information questionnaire, self-made left behind status questionnaire, self-made screen time questionnaire, Clancy autism behavior scale.EpiData 3.2 and SPSS 23.0 software were used for data entry and statistical analysis.Chi-square test was used to analyze the difference of positive rate of autism behavior.Logistic regression analysis was used to further explore the relationship between left-behind status, screen time and autism behavior of rural preschool children.Results:Compared with non-left-behind children (NLBC), the risk of autism behavior for left-behind children (LBC) increased 36%.The risk of autism behavior increased by 40% for 1 h/d 2 h/d when compared with ST ≤ 1 h/d.While comparing to NLBC with ST ≤ 1 h/d, the risk of autism behavior increased by 97% in LBC with 1 h/d 2 h/d.Conclusions:There is an additive effect on the risk of autism behavior when left-behind experience and excessive ST combined together.The daily ST should be strictly controlled within 2 h/d for NLBC, and within 1 h/d for LBC in order to reduce the risk of autism behavior in preschool children of rural areas.