Animals ; Anticholesteremic Agents ; pharmacology ; Cholesterol ; blood ; Drug Evaluation, Preclinical ; methods ; Male ; Rats

OBJECTIVEThe purpose of this research was to study the genetic diversity of F-ATPase subunit gene uncEBF derived from Streptococcus mutans (S. mutans) clinical isolates, furthermore to investigate the relationship between the genetic diversity of F-ATPase and S. mutans aciduric ability.

METHODS38 S. mutans strains included 18 high acid tolerance strains and 20 low acid tolerance strains. Gene uncEBF of these isolates were amplified with specific primers from S. mutans genomic DNA, and the PCR products were analyzed by RFLP and sequenced. SPSS 11.0 statistic software assayed the results.

RESULTSIt was testified that two genotypes A and B of PCR-RFLP were revealed when digested with Alu I and Dde I digested fragments of uncEBF displayed two different patterns C and D. Fisher exact two-tail test showed that the distributions of A and B genotype strains with different acidurance were different (P < 0.05), and the proportion of A genotype strains from high acidurance group was higher than that from low acidurance one. Some of these amplified uncEBF genes from different genotype were sequenced and testified that there existed variation of Alu I and Dde I recognized sites.

CONCLUSIONThis study indicated that uncEBF gene of S. mutans F-ATPase obviously exhibited genetic diversity.

Adenosine Triphosphatases ; Dental Caries ; Genetic Variation ; Genotype ; Humans ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Streptococcus mutans

OBJECTIVETo study the genetic diversity and the gene expression of membrane-bound proton-translocating ATPase (F-ATPase) subunit gene uncG derived from Streptococcus mutans (S. mutans) clinical isolates.

METHODS38 S. mutans strains derived from caries-active and caries-free individuals including 18 strains displaying high acid tolerance and 20 strains displaying low acid tolerance. Gene uncG was amplified with specific primers from S. mutans genomic DNA, then the PCR product was analyzed by RFLP and sequenced. The relative expression quantity of uncG gene against the housekeeping gene recA was determined by using RT-PCR method. A gel documentation system and QUANTITY ONES software were used to analyze the data results.

RESULTSIt was testified that four genotypes A, B, C and D of PCR-RFLP were revealed when respectively digested with Alu I and Bsr I, but the distributions of the four genotype strains showed no difference (P > 0.05). The differences of uncG gene transcript quantities derived from different genotype or different aciduranc strains had no significance (P > 0.05).

CONCLUSIONThis study indicated that uncG gene of F-ATPase obviously displayed genetic diversity and existed polymorphism at mRNA expression level, while the Alu I-RFLP genotypes and the expression levels would not be responsive to different acid tolerance of S. mutans strains.

Adenosine Triphosphatases ; Dental Caries ; Genetic Variation ; Genotype ; Humans ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Polymorphism, Restriction Fragment Length ; RNA, Messenger ; Streptococcus mutans

OBJECTIVETo explore a novel non-invasive method in detection of thyroid cancer by Fourier transform infrared (FTIR) spectroscopy.

METHODSSurface FTIR spectra of 15 cases of thyroid cancer and 51 cases of normal subjects were collected. 22 variables of 11 bands including peak positions and relative intensities were measured and all data were statistically analyzed.

RESULTSIn the cancer group: (1) the peak position of 1743 cm(-1) was shifted toward higher wave number (P < 0.05), and that of 1250 cm(-1) to the lower (P < 0.05), when compared to those of normal ones. (2) The relative intensity ratios of I(1546)/I(1460), I(1250)/I(1460), I(1120)/I(1460), I(1080)/I(1460) were significantly increased (P < 0.05). (3) The presence rate of band of 1340 cm(-1) was significantly decreased (P < 0.05).

CONCLUSIONFTIR surface spectra may become a novel powerful non-invasive approach of detecting thyroid cancer in regular routine check-up.

Humans ; Spectroscopy, Fourier Transform Infrared ; methods ; Thyroid Gland ; chemistry ; Thyroid Neoplasms ; chemistry ; diagnosis

OBJECTIVETo study hepatitis B virus (HBV) genotype and subtype distribution and its clinical significance in HBV-infected patients.

METHODSWe used type/subtype-specific primers and PCR to detect HBV genotype and subtype of 445 HBV-infected patients from Beijing, Changchun, Hanchuan Shenzhen, Qingyuan and Nanjing, including 7 acute hepatitis (AH), 36 asymptomatic HBV carriers (ASC), 352 chronic hepatitis (CH), 28 liver cirrhosis (LC), and 22 hepatocellular carcinoma (HCC) cases. Genotyping results were confirmed by PCR product sequencing.

RESULTSAmong 445 HBV-infected patients, the proportions of genotype B, C, and B/C were 32.6% (145/445), 53.7% (239/445), and 13.7% (61/445), respectively. In genotype C, 13 (5.4%) were subtype C1, 135 (56.5%) were subtype C2, and the remaining 91 (38.1%) were neither C1 nor C2. In genotype B, 100 (69.0%) were subtype Ba, 25 (17.2%) subtype Bj, and the other 20 (13.8%) were neither Ba nor Bj. In genotype B/C, 15 (24.6%) were Ba/C2, 8 (13.1%) Bj/C2, 6 (9.8%) Ba/C1, 3 (4.9%) Bj/C1, 11 (18.0%) Ba/neither C1 nor C2, 7 (11.5%) Bj/neither C1 nor C2, and 6 (9.8%) neither Ba nor Bj/neither C1 nor C2, 2 (3.3%) neither Ba nor Bj/C1, 3 (4.9%) neither Ba nor Bj/C2. The HBV genotype and subtype distribution we found exhibited significant differences in the various clinical types of HBV infection tested, and showed that genotype C was predominant among patients with liver cirrhosis (78.6%) and hepatocellular carcinoma (86.4%) while genotype B was predominant in asymptomatic carriers (72.2%). In addition, genotype and subtype distribution showed no significant differences between male and female patients, but genotype and subtype distribution showed significant differences in patients positive or negative with HBeAg.

CONCLUSIONSubtypes Ba and C2 are predominant in patients with hepatitis B from these 6 cities, and genotype C may be associated with the development of liver cirrhosis and hepatocellular carcinoma.

Genotype ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; virology ; Humans ; Liver Cirrhosis ; virology ; Liver Neoplasms ; virology ; Polymerase Chain Reaction

Objective To evaluate the long-term immunogencity and effectiveness of live attenuated hepatitis A (HA) vaccine (H2 strain) after one dose injection, through a 15 years' follow up observation. Methods A total of 220 children with negative anti-HAV antibody (aged 1-3 y)were involved and followed up in Jiaojiang district, Taizhou city, Zhejiang province. Indicators would include seroconversion and geometric meantiter(GMT) levels after inoculation the vaccine with single dose at 2 m, 12 m, 6 years, 10 years and 15 years. Epidemiological observation was carried out within the 15 years to evaluate the relationship between vaccine coverage, the incidence of HA and the overall effectiveness. In the studied population, serum was tested by ELISA(calibrated by WHO international reference) and ABBOTT Axsym HAVAB mEIA. Results Seroconversion rates were found to be 98.6% and 81.3% after 2 months and 15 years of inoculation and slowly decreased. GMT level was 128 mIU/ml after 15 years, significantly higher than the required protective level of 20 mIU/ml,recommended by WHO experts. Effectiveness through the 15-year follow up program showed a significant correlation between vaccine coverage and incidence of HA in 1-15 years aged group (Kendall-Rank test, t =-0.931, P＜0.01). There was no HA case seen among the observed accumulated 236 413 person-year vaccines, compared to 4 HA cases discovered in the 27 206 personyear of the non-vaccinees. The overall protective rate reached 100%. Through a mass vaccination program on children, the whole population established an immune-defence to enable the incidence of HA decreased by 96.7%. Conclusion The long-term immunogencity and effectiveness of live attenuated hepatitis A vaccine (H2 strain) after one dose injection could last as long as 15 years.

OBJECTIVETo study the genetic diversity of F-ATPase alpha subunit gene uncA derived from Streptococcus mutans (S. mutans) clinical isolates and to investigate the relationship between the genetic diversity of acidurance factor and S. mutans aciduric ability, also and the cariogenicity.

METHODSSixty-four S. mutans strains derived from 34 caries-active individuals and 30 caries-free individuals, including 18 strains displaying high acid tolerance and 20 strains displaying low acid tolerance. Gene uncA was amplified with specific primers from S. mutans genomic DNA, then the PCR products were analyzed by RFLP and sequenced.

RESULTSTwo genotypes A and B of PCR-RFLP were revealed when digested with Hph I. Mbo II also produced two different pattern C and D. The distributions of A and B genotype strains with different caries-sensitivity groups were different (P < 0.05), and the proportion of A genotype strains from caries-activity group was higher than that from caries-free one. The distributions of C and D genotype strains with different acidurance strains were different (P < 0.05), and the proportion of C genotype strains from high acid tolerance group was higher than that from low acid tolerance group. These amplified uncA genes from different group were sequenced and there existed variation of Hph I and Mbo II recognized sites.

CONCLUSIONSThis study indicates that uncA gene of S. mutans F-ATPase obviously displayed genetic diversity. The different Hph I-RFLP and Mbo II-RFLP genotypes could be related to the cariogenicity and acid tolerance of S. mutans strains.

Bacterial Proton-Translocating ATPases ; genetics ; Dental Caries ; microbiology ; Genes, Bacterial ; Genotype ; Humans ; Polymorphism, Restriction Fragment Length ; Streptococcus mutans ; enzymology ; genetics

BACKGROUNDLittle attention has been paid to the expression of heat shock protein 27 (HSP27) in patients with reflux esophagitis (RE), and few studies of the importance of HSP27 in esophagitis have been carried out in animal models. This study aimed to explore the expression of HSP27 in the esophageal tissue of rats with RE.

METHODSEighty female Wistar rats were randomly divided into experimental groups A and B and control groups C and D (n = 20 in each group). To establish RE, rats in the two experimental groups received pylorus and forestomach ligations, while rats in the control group received gastrostomy and gastric perforation repair. The rats in groups A and C were sacrificed 7 days after surgery, and the rats in groups B and D were sacrificed 14 days after surgery. In groups A and B, 10 and 8 rats were diagnosed with RE by pathological examination, respectively (they were included in groups A' and B', respectively). The histopathological diagnosis of all the lower esophageal tissues in groups C and D was normal and 20 normal specimens were randomly selected for groups C' and D' with 10 specimens in each group. Macroscopic and microscopic esophagitis scores were assessed for the specimens in groups A' and B'. Lower esophageal tissues were collected from groups A', B', C', and D', and paraffin-embedded slices were made using part of the tissues. The expression of HSP27 in the tissues was detected using the two-step streptavidin-peroxidase immunohistochemical method. Some collected tissues were frozen, and expressions of HSP27 mRNA were detected using fluorescence quantitative polymerase chain reaction (FQ-PCR).

RESULTSMedian macroscopic and microscopic esophagitis scores in groups A' (n = 10) and B' (n = 8) were 1.0 and 1.5, and 2.0 and 2.5, respectively. There were no significant differences in the macroscopic or microscopic esophagitis scores between the two groups (Z = -0.330, P = 0.741; Z = -0.142, P = 0.887, respectively). Immunohistochemical staining showed that HSP27 was expressed in all layers of the esophageal epithelia in RE and control rats. FQ-PCR showed that HSP27 mRNA levels in the lower esophageal tissue in RE group (groups A' and B') were higher than those in control group (groups C' and D') (Z = -0.249, P = 0.001). HSP27 mRNA expression in the lower esophageal tissue was significantly different in groups B' and D' (Z = -3.027, P = 0.002). And the levels of HSP27 mRNA expression in severe RE group (microscopic esophagitis score: 3) were higher than in mild RE group (microscopic esophagitis score: 1-2) and control group (Z = -3.396, P = 0.001; Z = -3.855, P < 0.001).

CONCLUSIONSHSP27 mRNA expression in the lower esophageal tissue of rats with RE is significantly higher than in the normal controls. Although reflux is a persistent stimulating factor, increased expression of HSP27 in the lower esophageal tissue of rats with RE requires aggravated esophageal injury.

Animals ; Esophagitis, Peptic ; metabolism ; Esophagus ; metabolism ; pathology ; Female ; HSP27 Heat-Shock Proteins ; genetics ; metabolism ; Immunohistochemistry ; Polymerase Chain Reaction ; Rats ; Rats, Wistar

Objective This study aims to investigate the frequency and characteristics of vascular involvement in Beh(c)et's disease (BD).Methods We enrolled patients who were hospitalized at Changhai Hospital affiliated Naval Medical University and who had been diagnosed with BD at discharge from January 1999 to July 2017.Patientswere divided into two groups,Vascular Beh(c)et's disease (VBD) group and non-VBD group,according to vascular involvement or not.We recorded and compared the demographicinformation,disease activity scores Beh(c)et’s Disease Current Activity Form (BDCAF),other systemic involvement and laboratory test results between the two groups.The clinical features of vascular involvement were analyzed.The measureddata were statistically analyzed with Student's t/t’ test or Mann-Whitney U test.The numerical data were statistically analyzed with x2 test or continuity correction x2 test.Results The total numbers of BD patients were 224,including 120 males and 104 females.Vascular involvements were found in 41 (18.3％) cases,including 34 males and 7 females.VBD was more common in males (28.3％ vs 6.7％,x2=17.388,P＜0.01).Of 41 VBD patients,11 cases (26.8％) had single vascular lesions,and 30 cases (73.2％) had multiple vascular lesions.Moreover,24 cases (58.5％) had arterial lesions,and 25 cases (61.0％) had venous lesions.Eight (19.5％) patients had both arterial and venous lesions.Compared to the non-VBD group,the VBD group had a higher frequency of cardiac involvement (22.0％ vs 3.8％,x2=16.592,P＜0.01),a higher disease activity index score (BDCAF) [3.0(2.0,4.0) score vs 2.0(2.0,3.0) score,U=2 609.5,P=0.001],higher levels of C-reactiveprotein (CRP) (14.45(4.97,64.08) mg/L vs 9.02(3.16,26.53) mg/L,U=2 809,P=0.046) and plasma homocysteine (Hcy) [(17.6±7.7) μmol/L vs (7.1±2.1) μmol/L,t'=7.894,P＜0.01].Conclusion VBD mainly affectsmales.It mainly presentsas multiple lesions.Moreover,patients in the VBD group havehigher frequency of cardiac complications,higher disease activity index scores (BDCAF),higher levels of CRP and Hcy than the non-VBD group.These results may have great valueto predict and diagnoseof VBD.

OBJECTIVE: To study the long-term prognosis of vesicoureteral reflux in children. METHODS: A retrospective analysis was performed for the clinical data of 138 children (218 ureters with reflux) who were diagnosed with vesicoureteral reflux for the first time from November 2005 to March 2017 and received medical treatment and regular follow-up. According to the initial grade of reflux, the ureters with reflux were divided into a low-grade group (141 ureters, grade I-III) and a high-grade group (77 ureters, grade IV-V), and the two groups were compared in terms of clinical data and follow-up results. RESULTS: Among the 138 children, there were 82 boys and 56 girls. Their age ranged from 31 days to 10 years at the time of confirmed diagnosis, with a median age of 8 months. The follow-up time ranged from 8 months to 7 years, with a median follow-up time of 1.4 years. At the time of confirmed diagnosis, the high-grade group had significantly higher levels of urinary N-acetyl-β-D-glucosaminidase, urinary microalbumin and urinary immunoglobulin G than the low-grade group (P<0.05). Among the 218 ureters with reflux, 132 (60.6%) achieved a reduction in the grade of reflux (among which 74 achieved complete disappearance of reflux), 43 had no change in the grade of reflux, and 43 had an increase in the grade of reflux. Compared with the high-grade group, the low-grade group had a significantly higher complete remission rate (41.1% vs 19.5%, P=0.001) and a significantly shorter time to complete remission of reflux (P=0.002). CONCLUSIONS Most children with vesicoureteral reflux can achieve a reduction in the grade of reflux or even complete disappearance of reflux during follow-up, and the children with low-grade reflux have better prognosis than those with high-grade reflux.
Child ; Child, Preschool ; Female ; Follow-Up Studies ; Humans ; Infant ; Male ; Prognosis ; Retrospective Studies ; Vesico-Ureteral Reflux