Objective:To determine the molecular role of DNA methyltransferase (DNMT) in kidney tumorigenesis. Methods:Tissue samples consisted of 15 cancer tissues and 15 matched adjacent tissues from clear cell renal cell carcinoma (ccRCC) patients who had undergone nephrectomy in 2012 at the First Affiliated Hospital of Baotou Medical College, Science and Technology University of Inner Mongolia were collected. Real-time PCR, Western blot, combined bisulfite restriction analysis (COBRA) and methylation specific PCR were used in this study. Real-time PCR was used to examine the mRNA expression levels of DNMT. The global methylation level, DNA methylation level, and the expression of the antioncogene RASSF1A in ccRCC tissues were concurrently detected. Results:Both mRNA and protein levels of DNMT3B4, a splice variant of DNMT3B, were elevated in renal cell carcinoma tissue compared with those in con-troll tissue. Additionally, Alu was hypomethylated in ccRCC tissue (0.106±0.04) compared with control tissue (0.115±0.03) (P<0.05). Fur-thermore, the methylation of the promoter for RASSF1A, a tumor-suppressor gene, moderately increased in renal cell carcinoma tis-sue. By contrast, RASF1A expression decreased. Conclusion:DNMT3B4 overexpression may play an important role in human kidney tu-morigenesis via chromosomal instability and the decreased expression of RASSF1A.

Objective To determine Neisseria gonorrhoeae multi-antigen sequence typing (NG-MAST) sequence types in different geographical areas of China,including Changzhou and Yangzhou cities of Jiangsu province,Wuzhou and Hezhou cities in Guangxi Zhuang Autonomous region,Sanya and Qionghai cities of Hainan province,Jiangmen and Maoming cities of Guangdong province.Methods DNA was extracted using Qiagen DX extraction kits from 88 urine samples which were collected from male patients attending sexually transmitted disease (STD) clinics and positive for nucleic acid amplification test (NAAT) for N.gonorrhoeae.Two rounds of PCR were carried out to amplify the porB and tbpB genes of N.gonorrhoeae followed by gene sequencing.Sequence alignment was performed on the NG-MAST website (http://www.ng-mast.net) to determine the genotype of N.gonorrhoeae.Results The first-round PCR yielded positive results for porB and tbpB in 13.6％ (12/88) and 14.8％ (13/88),respectively,of these urine specimens,and 12 samples were successfully genotyped with the efficiency of genotyping being 13.6％.The amplification efficiency of second-round PCR was enhanced to 71.6％ and 72.7％ for porB and tbpB,respectively,and the efficiency of genotyping increased to 70.5％ (62/88).Compared with the first-round PCR,the second-round PCR showed an increase in amplification efficiency for porB and tbpB by 58.0％ and 57.9％ respectively,as well as in genotyping efficiency by 56.9％.Forty-five genotypes were identified in the 62 samples,including 40 known genotypes and 5 novel genotypes.Of these genotypes,ST1866 was the most abundant (6/62),followed by ST1972 (4/62) and ST3356 (4/62),all of which were from Jiangsu province.The ST532 genotype was identified in 3 samples from Guangdong province,ST2221 genotype in 2 samples from Guangxi Zhuang Autonomous region.Each of the remaining genotypes was identified in only 1 sample and scattered in all of these cities.The 5 novel MAST-genotypes were as follows:porB-892 and tbpB-46 (98％ similarity),porB-130 and tbpB-504 (96％ similarity),porB-2790 and tbpB-32 (99％ similarity),porB-1053 and tbpB-856 (99％ similarity).Conclusions Urine samples can be used for NG-MAST analysis,and two rounds of PCR can enhance the efficiency of genotyping.NG-MAST genotypes appear to be diverse in different geographical areas of China.

Objective To assess the prevalence of urogenital infection with and genotype distribution of C.trachomatis among female sex workers (FSWs) from different entertainment venues in Wuzhou and Hezhou cities of Guangxi Zhuang Autonomous Region.Methods A total of 810 FSWs were recruited to this study by convenience sampling from entertainment venues in Wuzhou and Hezhou cities of Guangxi Zhuang Autonomous Region from July 2009 to September 2010.Based on the venues where they solicited clients,the FSWs were classified into three tiers,i.e.,high-tier,middle-tier and low-tier.Cervical swabs were collected from all of these subjects followed by detection of C.trachomatis with the Amplicor PCR test kit.Then,DNA was extracted from C.trachomatis-positive specimens and subjected to nested PCR assay targeting the ompA gene followed by bidirectional sequencing.The genotype of C.trachomatis was determined according to the sequence of ompA gene.Chi-square test was conducted to compare the urogenital infection rate and genotype distribution of C.trachomatis between different tiers of FSWs.Results Among the 805 FSWs,the prevalence rate of urogenital C.trachomatis infection was 20.0％ (161/805).Chi-square test showed that the prevalence rate of urogenital C.trachomatis infection was significantly lower in high-and middle-tier FSWs than in low-tier FSWs (x2 =3.97,5.95,respectively,both P ＜ 0.05).Nine genotypes of C.trachomatis were identified in these FSWs,with serotype F as the most prevalent genotype (39/154,25.3％).Low-tier FSWs showed a higher frequency of genotype E (x2 =5.02,P ＜ 0.05) but a lower frequency of genotype K (Fisher's Exact test,P =0.048) compared with middle-tier FSWs.Conclusions Low-tier FSWs show a high rate of urogenital infection with C.trachomatis,with serotype E as the prevalent type.Since C.trachomatis serovar E-infected patients are likely to be missed by symptom-based screening and preventive strategies,standardized screening for and efficient treatment of urogenital C.trachomatis infection should be enhanced among low-tier FSWs for the prevention of C.trachomatis transmission.

Objective To investigate in vitro combined effect of ceftriaxone and azithromycin against clinical isolates of Neisseria gonorrhoeae(NG). Methods A total of 25 NG clinical isolates were collected from the STD clinic of Dalian Dermatosis Hospital in 2012. Epsilometer test(Etest)method was used to determine the minimum inhibitory concentrations(MICs)of ceftriaxone and azithromycin against NG isolates. Fractional inhibitory concentration index (FICI) was calculated to evaluate the in vitro combined effect of ceftriaxone and azithromycin against NG isolates. Results The mean MICs of ceftriaxone and azithromycin were 0.032 mg/L (range, 0.008- 0.064 mg/L) and 0.834 mg/L (range, 0.064-4.000 mg/L), respectively. The FICI ranged from 0.724 to 2.696, and ceftriaxone and azithromycin showed an additive effect against the above NG isolates. Conclusion Ceftriaxone and azithromycin show an additive effect against NG in vitro, but further studies with large sample size are needed to confirm their effects.

Objective To test the ceftriaxone susceptibility of Neisseria gonorrhoeae (NG) isolates from Nanjing city,and to assess their genotypes by using the NG multi-antigen sequence typing (NG-MAST) method.Methods A total of 204 NG strains isolated in 2007 and 81 in 2012 from Nanjing city were included in this study.The minimum inhibitory concentration (MIC) of ceftriaxone was determined for these strains using an agar dilution method.DNA was extracted by the Qiagen commercial kit from these strains followed by NG-MAST.Results All the isolates were susceptible to ceftriaxone (MIC,≤ 0.25 μg/ml).The MIC of ceftriaxone was ≥ 0.06 μg/ml for 63.2％ of all the NG strains,70.6％ of those isolated in 2007 and 44.4％ of those in 2012,and ≥ 0.125 μg/ml for 31.6 ％ of all the NG strains,39.7％ of those isolated in 2007,11.1％ of those in 2012.Totally,166 genotypes were identified among the 285 isolates,of which,73 had been reported,and 93 were previously unreported.The most prevalent genotype was ST568 (n =13) in NG strains isolated in 2007,followed by ST270 (n =9),ST421 (n =7),ST2288 (n =5),ST1731 (n =4),ST1766 (n =4),ST1866 (n =4),ST1870 (n =4),while ST2318 (n =5),ST1053 (n =4),ST5990 (n =4),ST8726 (n =4) were the common genotypes in 2012.Those isolates with identical or similar genotypes tended to display similar MICs for ceftriaxone.Conclusions The prevalent genotypes of NG are markedly different between 2007 and 2012 in Nanjing region,and there is a strong association between the genotypes and ceftriaxone susceptibility of NG.NG-MAST results may serve as a genetic marker in the surveillance of antibiotic susceptibility in NG.

Background and purpose:DNMT3B has nearly 40 known splice variants expressed in a tissue-and disease-speciifc manner, but the roles of these splice variants in the cell are still unclear. The aim of this study was to investigate the effects of overexpression of DNA methyltransferase 3B4 (DNMT3B4) gene on proliferation of human embryo kidney 293A cells. Methods:293A cells were transfected with plasmid pCMV-DNMT3B4 or pCMV-2B and then treated with G418 to get the stable cell line. The stable cell lines were determined for proliferation level by MTT method, and for cell cycle distribution by lfow cytometry. The expression of p21 was detected by real-time PCR and Western blot. The methylation status of p21 gene promoter was detected by methylation-speciifc PCR (MS-PCR). Results:The absorbance value in DNMT3B4-1 and DNMT3B4-2 clone were (58.92±3.47)%and (68.82±5.64)%as compared to 293A-vector cells using MTT method. DNMT3B4 overexpression signiifcantly decreased cell proliferation (P<0.05). S phase fraction of 293A-vector cells was (40.44±0.91)%. While in DNMT3B4-1 and DNMT3B4-2 clone cells, the S phase fraction was (35.88±2.00)%and (37.00±1.79)%respectively. Overexpression of DNMT3B4 could significantly decrease S phase fraction (P<0.05). The expression of p21 in DNMT3B4 overexpressed cells was increased, but the methylation status of p21 gene promoter was unchanged.Conclusion:Overexpression of DNMT3B4 can inhibit 293A cell proliferation and can facilitate p21 expression.

Objective:To compare the application value of different polypropylene mesh in inguinal hernia repair of adults.Methods:The prospective cohort study was conducted. The clinical data of 120 adult patients with inguinal hernia who were admitted to two medical centers (60 in Huadong Hospital affiliated to Fudan University and 60 in Shanghai Ninth People′s Hospital affiliated to Shanghai Jiaotong University from March 2012 to Match 2014 were collected. Patients were randomly divided into study group and control group using the random number table. Patients in the study group underwent repair of inguinal hernia using the SMP95958X mesh, and patients in the control group underwent repair of inguinal hernia using the modified Kugel mesh. All patients underwent preperitoneal repair by senior hernia surgery specialists. Observation indicators: (1) postoperative pain; (2) complications and follow-up. Patients were followed up at postoperative 3 months and 6 months using outpatient examination to detect the short-term complications by physical or color doppler ultrasonography examination, and at postoperative 5 years using telephone interview or outpatient examination to detect long-term complications including infection, foreign body sensation and recurrence of hernia. The follow-up was up to March 2019. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups was analyzed using the t test. Measurement data with skewed distribution were represented as M (range), comparison between groups was analyzed using the nonparametric rank sum test. Count data were described as absolute numbers or percentages, and comparison between groups was analyzed using the chi-square, continuous correction chi-square test or Fisher exact probability. Comparison of ranked data were analyzed using the nonparametric rank sum test. Results:A total of 118 patients with inguinal hernia were selected for eligibility, including 116 males and 2 females, aged (64±12)years, with a range from 29 to 84 years. Of the 118 patients, 59 were in the study group and 59 were in the control group, respectively. (1) Postoperative pain: of the 59 patients in the control group, 34 took painkiller and 1 case lost the data of taking painkiller at postoperative 2 days. Of the 59 patients in the study group, 29 cases took painkiller. There was no significant difference in taking painkiller between the two groups ( χ2=1.055, P>0.05). The pain score at postoperative 2 days and 3 months were 3.26(range, 0.70-6.90) and 0.87(range, 0.00-4.10) of the control group, respectively, and 3.03(range, 0.00-8.80) and 0.83(range, 0.00-3.10) of the study group, respectively, showing no significant difference between the two groups ( Z=0.782, 0.729, P>0.05). (2) Complications and follow-up: the incidence of postoperative complications at perioperative period (within postoperative 2 days) was 1.7%(1/59) and 1.7%(1/59) in the control group and study group, respectively, showing no significant difference between the two groups ( P>0.05). Both of 59 patients in the control group and study group were followed up for 6 months after operation, respectively. The incidence of postoperative complications at 3 months and 6 months after operation was 1.7%(1/59) and 1.7%(1/59) in the control group, respectively, and 5.1%(3/59) and 5.1%(3/59) in the study group, showing no significant difference between the two groups ( P>0.05). Fifty-five patients of the control group and 52 patients of the study group were followed up for 5 years after operation, respectively. There was 1 case of infection in the control group, with the incidence of postoperative long-term ( within 5 years after operation) complication of 1.8%(1/55), and there were 2 cases of infection and 1 case of foreign body sensation in the study group with the incidence of postoperative long-term (within 5 years after operation) complication of 5.8%(3/52), showing no significant difference between the two group ( P>0.05). There was no recurrence of hernia in either group. Conclusion:Both of the SMP95958X mesh and the modified Kugel mesh can be used in preperitoneal repair of inguinal hernia, showing no significant difference in the efficacy between them.

Objective: Based on observational, longitudinal and intervention study of common diseases among students in Jiangsu Province, this paper presents the current progress of two year follow up of myopia cohort regarding the association between growth parameters with progression of myopia among children and adolescents in areas with rapid economic growth. Methods: This survey adopted the stratified cluster sampling method for school selection. The full automatic computer optometry (TOPCON RM800) was used to track myopia related parameters for all participants from 2019 to 2020 under the condition of mydriasis (compound topicamide eye drops). Relationship between growth parameters of children and adolescents and the incidence and progression of myopia was analyzed by using Cox regression multiple statistical model. Results: The myopia rates of students from grade 1 to grade 3 in 2019 were 5.4%, 21.5% and 37.3% respectively. After one year, the myopia rates of all school stages increased to 25.3%, 43.3% and 58.1% respectively( χ 2=53.59, 49.63, 32.52, P <0.01). The mean diopter of right eye and left eye after mydriasis were ( 0.30± 1.24/0.39±1.26)D in 2019 and (-0.33±1.54/-0.19±1.55)D in 2020, respectively based on Cox multiple regression results, age ( HR =1.21, 95% CI =1.09-1.34), naked eye vision ( HR =0.08, 95% CI =0.07-0.11), height ( HR =0.98, 95% CI =0.97-0.99) showed a strong correlation with the incidence and progression of myopia( P <0.05). Conclusion Myopia is growing rapidly in the central region of Jiangsu Province. It is suggested that diopter, axial length, naked eye vision, age, height and other indicators should be included in the refractive archives of children and adolescents in the region.