Abstract:Objective:TSLC26A4 gene is thought to be the second major contributor to hereditary hearing loss. At least 170 pathological mutations of SLC26A4 were identified in deafness patients,and showed regional differences and ethnic specificity for different groups around the world. However,no unanimous conclusion was achieved on the high frequency mutations of SLC26A4 which induced the hearing loss. This study is aimed to invistigate the molecular structure and racial difference of high frequency pathological SLC26A4 mutations in hearing loss. Methods: Total of 662 published epidemiological studies of SLC26A4 mutation and deafness was collected from 1997,Jan to 2014,Dec. through the databases. Based on the NOS standard,31 of the articles were included. meta-analysis was carried to study the data of SLC26A4 mutation frequency. Stata11.2 and RevMan 5.1 software was used to describe the data of literature for meta-analysis to explore deaf risk correlation. Swiss Model software was used to analyze the molecular structure of high frequency pathological SLC26A4 mutations. Results: ①A variety of types of SLC26A4 mutations are associated with an increased deafness risk(OR= 39.73,95% CI: 21.359-73.903,P < 0.0001);② It indicates the significant heterogeneity in Asian but not in European and American;③Particularly,the 6 kinds of mutations were calculated as high frequency pathological SLC26A4 mutations (p.V138F,p.G209V,IVS7-2A>G,IVS8+1G>A,p.T416P and p.H723R),protein structure changes of which were simulated by Swiss Model. Conclusion: These findings advance our knowledge of the genetic basis of SCL26A4 variation with hereditary hearing loss in the multiethnic populations

Abstract:Objective:To study the effects of adenovirus mediated prostate apoptosis response-4 (Par-4)combined with cisplatin(CDDP)on subcutaneous xenografts of hunan Wilms’ tumor cells SK-NEP-1 in nude mouse. Methods:Inhibition effect of subcutaneous transplanted Wilms’ tumor model were established in BALB/c nude mice,and then the nude mice were randomly divided into five groups,namely Par-4+CDDP group,Par-4 group,CDDP group,negative control Par-4 group and blank control group. The volumes and weights of the transplanted tumors were measured. Transplanted tumor tissues were used to detect the expression levels of Par-4,GRP78 and BAX protein by HE staining and immunohistochemical analysis. Results:The tumor growth was inhibited in Par-4+CDDP group,Par-4 group and CDDP group,and there were differences in the tumor weight amorg 3 groups (P < 0.05). The inhibition effect of Par-4+CDDP group was superior to the Par-4 group and CDDP group. There was no statistically significace in tumor inhibition between blank control group and negative control Par-4 group. Par-4+CDDP can significantly increased Par-4,GRP78 and BAX protein(P < 0.05). Conclusion: Ectopic Par-4 in combination with CDDP suppressed the cancer-xenografted growth in nude mice,which might up-regulate BAX protein,the result of the interraction of extracellular Par-4 and GRP78.

Abstract:Objective:To determine target genes of transcription factors THAP1 for a new therapeutic strategy of neonatal lung injury. Methods:Chromatin DNA in alveolar type Ⅱ epithelial cells was immunoprecipitated with THAP1 antibody. The obtained DNA samples were then subjected to high-throughput sequencing and bioinformatic analysis. Results:A total 20 417 enrichment peaks corresponding 9 688 genes was found. The mapping of the peaks revealed that the targeted nucleotide sequences located mainly within intergenic regions,intron regions and promoter regions,which contained 1 051 gene promoter regions. Gene Ontology(GO)analysis suggested that THAP1 regulated a set of genes,which were involved in various biological processes including protein dimerization activity,identical protein binding,protein phosphatase regulator activity,steroid hormone receptor activity. Signal pathway analysis showed that these genes were associated with endocytosis,glycan degradation,and biosynthesis-chondroitin sulfate/dermatan sulfate. Conclusion:The targeted genes of the transcription factor THAP1 in the alveolar type Ⅱ epithelial cells were determined. These results show that THAP1 is widely involved in various biological processes,which might provide a useful insight to reveal the function and mechanism of THAP1 gene on neonatal lung injury.

Abstract:Objective:To observe the effect of stromal cell derived factor-1 (SDF-1)on human microvascular endothelial cell line-1(HMEC-1),and study the relevant mechanism. Methods:HMEC-1 cells were treated with SDF-1 of different concentrations(0,25,50,100 -滋g/L),then the phosphorylation level of PI3K/Akt and MAPK/Erk were detected by Western blot,and the proliferation ability and apoptosis rate of HMEC-1 cells were detected by MTT and FCM. Changes on migration ability of HMEC-1 cells was detected by wound healing assay. MCF-7 cells were pretreated with PI3K/Akt and MAPK/Erk signaling pathway inhibitor to observe the effect of PI3K/Akt and MAPK/Erk signaling pathway on SDF-1 induced proliferation,apoptosis and migration in HMEC-1 cells. Results:Compared with 0 -滋g/L SDF-1 treated group,25,50 and 100 -滋g/L SDF-1 treatment actived PI3K/Akt and MAPK/Erk signaling pathway,promoted proliferation and migration ability,and induced HMEC-1 cells apoptosis(P < 0.01),and those effects were concentration-dependent. When pretreated with PI3K/Akt and MAPK/Erk signaling pathway inhibitor,the effect of SDF-1 on promoting proliferation and migration ability,and inducing HMEC-1 cells apoptosis ability were significantly blocked. Conclusion:By activation PI3K/Akt and MAPK/Erk signaling pathway,SDF-1 can promote proliferation and migration ability,and induce HMEC-1 cells apoptosis,thus plays a role in diabetic angiopathy.

Abstract:Objective:We established a HepG2 hepatic steatosis model to observe whether liraglutide improves lipid metabolism in HepG2 liver cells,and discuss the related mechanisms. Methods:Sodium palmitate was performed to induce HepG2 steatosis model,and liraglutide intervention was given. Oil Red O staining was performed to determine the establishment of HepG2 hepatic steatosis model. Key enzyme of lipid synthesis and degradation as well as the activation of PI3K signaling pathway in HepG2 liver cells were detected by Western blotting assay. To observe the effect of PI3K signaling pathway in sodium palmitate induced HepG2 hepatic steatosis,HepG2 liver cells was pretreated with PI3K signaling pathway inhibitor. Results:Oil Red O staining showed that the model was successfully established. Western blotting assay showed that sodium palmitate significantly increased the expression of sterol regulatory element-binding protein1c(SREBP1c)and fatty acid synthase(FAS)protein levels in HepG2 liver cells,and decreased adipose triglyceride lipase (ATGL)protein levels(P < 0.01); sodium palmitate activated PI3K signaling pathway in HepG2 liver cells. Compared with sodium palmitate,liraglutide significantly decreased the expression of SREBP1c and FAS protein levels in HepG2 liver cells,and increased ATGL protein levels; liraglutide inhibited PI3K signaling pathway in HepG2 liver cells. After blocking PI3K signaling pathway,sodium palmitate-induced steatosis of HepG2 liver cells was significantly reduced (P < 0.01). Conclusion:By regulating PI3K signaling pathway in HepG2 liver cells,liraglutide improves lipid metabolism in HepG2 liver cells.

Abstract:Objective:To investigate the relationship between serum pigment epithelium-derived factor (PEDF) level,PEDF gene polymorphism in promoter region rs1294385 and microalbuminuria in type 2 diabetes mellitus (T2DM)of Fujian Han population. Methods:According to urinary albumin to creatinine ratio (UACR),471 T2DM patients were divided into the normal albuminuria group(the NAU group,UACR<30 μg/mg,n=246) and the microalbuminuria group(the MAU group,30 μg/mg≤UACR<300 μg/mg,n=225). The PEDF gene rs1294385 G→A polymorphism was measured based on polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP). Meanwhile,serum PEDF level,fasting plasma glucose(FPG),fasting insulin(FINS),glycosylated hemoglobin (HbA1c) and blood lipid were measured. Results:The level of serum PEDF in the MAU group was significantly higher than that in the NAU group(P < 0.05). According to partial correlation analysis,serum PEDF level was positively related with UACR(P < 0.05). There were significant differences between the frequencies of PEDF gene rs1294385G→A genotype (GG genotype,GA genotype and AA genotype)and allele(G/A)of the NAU group and those of the MAU group(P < 0.05). The risk ratio of GA genotype carriers suffering microalbuminuria was 1.838 times higher than that of GG genotype,while the risk ratio of GA and AA genotype carriers suffering microalbuminuria was 1.862 times higher than that of GG genotype(P < 0.05). Conclusion:Elevation of serum PEDF level is in accordance with the development of microalbuminuria in T2DM patients. Gene polymorphism in promoter region rs1294385 G→A of PEDF gene is closely related with microalbuminuria in T2DM patients of Fujian Han population. Moreover,T2DM patients with carriers of the A allele gene may have a higher risk of suffering from microalbuminuria.

Abstract:Objective:To explore the function and regulation mechanism of dopamine(DA)and cocaine-and amphetamine-regulated transcript peptides (CART)in ischemic stroke. Methods:The acute ischemic stroke model in rat was established,accompanied with administration of DA and CART. The neurological deficit scores,infarct volume,and inflammasome activity were detected to clarify the role of DA and CART in acute ischemic. Results:DA and CART reduced neurological deficit scores and infarct volumin in rats with acute ischemic stroke,and inhibited the activation of the inflammasome by reducing Caspase-1 and IL-1β secretion. Moreover,the expression of CART is regulated by DA. Conclusion:Dopamine can reducing inflammasome activity to protect ischemic stroke by regulation of CART.

Abstract:Objective:The aim of this study was to investigate the expression of Tim-3 in pathogenesis of IgA nephropathy(IgAN)and its clinical significance. Methods:Real time qPCR was performed to detect Tim-3 mRNA in peripheral blood mononuclear cells(PBMCs)from 41 IgAN patients and 21 normal subjects. The proportion of Tim-3+ cells was assayed by flow cytometry. Results:24-hour proteinuria,serum creatinine,routine urine test,urine sediment examination and red blood cell count at high magnification were recorded before renal biopsy. Results:Tim-3 mRNA expression was increased significantly in IgAN patients compared with that in healthy controls(0.64 ± 0.27 vs. 0.18 ± 0.09,P < 0.01),and there was a nearly positive correlation between pathological grade and expression degree of Tim-3 mRNA. Levels of Tim-3 mRNA expression in patients with grade Ⅲ,Ⅳ and Ⅴ were higher than those in patients with grade Ⅰ and Ⅱ(P < 0.05). Also,we found that the proportion of Tim-3+ cells in PBMCs from IgAN patients was higher than that from normal controls[(1.94 ± 0.77)% vs. (0.59 ± 0.24)%,P < 0.01]. The proportion of Tim-3+ cells in patients with grade Ⅳ and Ⅴ was higher than that in patients with grade Ⅰ,Ⅱ and Ⅲ(P < 0.05). Moreover,the expression of Tim-3 mRNA and the proportion of Tim-3+ cells were significantly positively related to 24-hour proteinuria and serum creatinine. Conclusion:Up-regulation of Tim-3 in IgAN patients was positively correlated with some clinical indexes including 24-hour proteinuria and serum creatinine. Tim-3 may be involved in the pathogenesis of IgAN,and can be a reliable biomarker for evaluating disease severity and prognosis of IgAN.

Abstract:Objective:To investigated the correlation between galectin 3 and chronic heart failure classification,treatment and prognosis. Methods:Total of 140 patients with chronic heart failure in our hospital were selected from January 2011 to January 2013,and divided into NYHA class Ⅱ group(47 cases),Ⅲ level group(50 cases),Ⅳ level group(43 cases). Serum galectin-3,high sensitive C-reactive protein(hs-CRP)and N-terminal pro-brain natriuretic peptide (NT-proBNP)were detected. Left ventricular end-diastolic dimension (LVEDD)and ejection fraction (EF)were also observed. Serum galectin-3 level was compared between patients with ineffective and effective treatment results. Correction between prognosis and galectin-3 was analyzed. Results:Galectin-3 was positively correlated with hs-CRP,lnNT-proBNP,LVEDD,respevtively (r=0.912,P < 0.05;r=0.772,P < 0.05;r=0.564,P < 0.05),and negative correlated with EF(r=-0.612,P < 0.05). Serum galectin-3 level of effective treatment group was significantly higher than that of ineffective treatment group(t = 3.81,P < 0.01). And serum galectin-3 level of group with cardiac events was significantly higher than that of group without cardiac events(t = 3.04,P < 0.01). Conclusion:Galectin-3 is closely related to chronic heart failure,and could be used for clinical diagnosis of chronic heart failure.

Abstract:Objective:To compare intraoperative use of gelatin and hydroxyethyl starch 130/0.4 in elderly severe patients undergoing coronary artery bypass surgery, in terms of volume expansion efficiency, side effects and clinical prognosis. Methods: Forty severe patients aged ≥60 years old undergoing coronary artery bypass surgery were randomized to receive intraoperative gelatin (n=20) or HES130/0.4 (n=20) as volume replacement. Amount of crystalloid and colloid solution, blood coagulation, bleeding, blood products infusion, serum creatinine, and time for tracheal tube extraction, length of stay in ICU and hospital were measured and compared. Results: Intraoperative amount of gelatin was slightly less than HES130/0.4 (P=0.35), while amount of crystalloid solution needed between groups was comparable. No significant difference of blood coagulation, bleeding and blood products infusion was found between groups. Serum creatinine increased transiently in both groups to a comparable extent on the 1st POD, and subsequently decreased to normal on the 4th and 7th POD, with no difference between groups at each time point. Time for tracheal tube extraction and length of stay in ICU did not differ significantly between groups, while hospital length of post-operative stay was longer but not significantly in Gelatin group compared to HES130/0.4 group. Conclusion: In elderly severe patients undergoing coronary artery bypass surgery, intraoperative volume therapy with gelatin or HES130/0.4 showed no significant difference between groups in terms of both volume expansion efficiency and side effects like blood coagulation and renal function.

Abstract:Objective:To evaluate whether the microbial culture results of fluid or tissue obtained from the affected joint in patients with chronic periprosthetic joint infection (PJI) after total hip arthroplasty (THA) affect the effect of two-stage revision surgery. Methods:A retrospective study including 48 patients who underwent two-stage revision surgery with chronic PJI after THA from General Hospital of Beijing Military Region was conducted. The first stage comprised prosthesis extraction,radical debridement and insertion of vancomycin-loaded cement hip spacer. After eradicating the infection,a cementless THA was implanted in the second stage. Postoperatively,all patients were received sensitive antibiotics via intravenous drip for 6 weeks. Patients were assessed for infection control time,infection recurrence,pain evaluation[visual analog scale(VAS)]and hip joint function evaluation(Harris Hip score). X-ray photograph was routinely performed. Results:The numbers of microorganism in the culture-negative (CN)group and bacteria culture-positive(CP)group were 13 and 35,respectively. After debridement revision in the first stage,infection was controlled in all patients. The time for controlling infection in the CN group was longer than that in the CP group[(119.3 ± 30.1)d vs. (98.4 ± 25.9)d,P=0.03]. The average duration of follow-up was 29.6 (range,9~49)months in outpatient department. At last follow-up,Harris hip score and pain VAS score were significantly improved compared with preoperation in all patients. Compared with the CP group,patients in the CN group were with a higher pain VAS score[2.8 ± 1.6 vs. (1.8 ± 1.2),P=0.03],a lower Harris function score[(79.1 ± 9.1) vs. (85.6 ± 7.1),P=0.01]as well as excellent and good rate[53.8% vs. 85.7%,P=0.04]. There were 2 cases of recurrence respectively in the CN group and CP group,but no statistical difference of recurrence rate was observed between the two groups[15.4% vs. 5.7%,P=0.62]. Conclusion:Satisfactory outcomes can be obtained with two-stage revision using vancomycin-loaded cement hip spacers and cementless implants for prosthetic hip joint infections. However,microbial culture results may affect the efficacy of two-stage revision in PJI patients,patients with a culture negative result may get a poor prognosis. The improvement of the detection rate of microoganism and application sensitive antibiotics may have a profound meaning on the effect of PJI.

Abstract:Objective:To investigate cytologic features,immunocytochemical profiles and differential diagnoses of solid-pseudopapillary neoplasms (SPN) of the pancreas in endoscopic ultrasound-guided fine needle aspiration (EUS-FNA)specimen. Methods:The cytopathological and immunocytochemical features were studied in 7 cases of SPN,who were diagnosed by EUS-FNA cytology and were confirmed by histopathology. Results:Samples of EUS-FNA gotten from SPN were usually highly cellular. Tumor cells were usually small and uniform,round-to-plasmacytoid,had pale-to-granular cytoplasm with vacuoles. The nuclei of neoplastic cells were usually round or orbicular-ovate,with smooth contours,fine and evenly dispersed granular chromatin. Nucleoli were not seen or inconspicuous. The neoplastic cells were presented as diffused distribution,rosette/acinar pattern,and more specifically,papillae with fibrovascular cores. Immunocytochemical staining was performed on 5 cases. Vimentin,progesterone receptor,β-catenin,CD10,CD56 and α-1-antitrypsin showed strong positivity in all 5 cases,and synaptophysin,chromogranin A,cytokeratin AE1/AE3 as well as CD99 variable positivity in some of the cases. Conclusion:Cyto-morphological characteristics of SPN includes small uniform tumor cells,which are disperse or arranged in papillae with fibrovascular cores. Differential diagnoses of SPN include pancreatic endocrine neoplasm and acinar cell carcinoma etc. The expression of β-catenin,CD10,CD56,vimentin,progesterone receptor and CD99 can promote the diagnosis of SPN.

Abstract:Objective:To invertigate the reproductive risk of chromosomal translocation carriers,in order to provide data for preconception counseling preimplantation genetic diagnosis (PGD). Methods:A total of 173 cases with chromosomal balanced translocation were divided into different groups based on the type of translocation,gender and breakpoint positions, and a total of the reproductive risk of each group was assessed. Results:①Tweenty-eight of 173 cases were infertility,and 145 cases had pregnancy history. Pregnancies were recorded in detais. The pregnancy outcomes were as follows:248 spontaneous abortions,4 induced terminations because of fetal abnormality,60 normal/balanced offsprings. The rates of spontaneous abortion of the reciprocal translocation carriers was significantly higher than that of Robertsonian translocation carriers (87.3% vs 64.5%,P < 0.05);②Among 96 reciprocal translocation carriers with pregnancy history,the incidence of spontaneous abortion in translocations with acrocentric chromosomes was higher than those without acrocentric chromosomes (92.7% vs 81.2%,P < 0.05). The incidence of spontaneous abortion in translocations with terminal breakpoints was higher than those without terminal breakpoints (95.5% vs 81.4%,P < 0.05);③Among 58 male carriers,18 cases was infertility and 35 had pregnancy history. Among 120 female carriers,10 was infertility and 110 had pregnancy history. The incidence of infertility of female carries was higher than that of the males(34.0% vs 8.3%,P <0.05). The rates of spontaneous abortion of the female carriers with pregnancy history was significantly higher than that of the male carriers(83.7% vs 71.2%,P < 0.05). Conclusion:①Balanced translocation carriers suffer from poor pregnancy prognosis,and their most common pregnancy outcome is recurrent miscarriage in early pregnancy;②The reproductive risk of reciprocal translocation carriers was significantly higher than that of Robertsonian translocation carriers;③The reproductive risk of reciprocal translocations carriers with acrocentric chromosomes and terminal breakpoints was higher than that of those without acrocentric chromosomes and terminal breakpoints;④The reproductive risk of the female translocation carriers was significantly higher than that of male translocation carriers. ⑤Balanced tranalocations markedly affect male fertility but no significant affect on conception of female.

Abstract:Objective:To analyze twin clinical reproduction outcome after in vitro fertilization (IVF)/intracytoplasmic sperm injection(ICSI)-embryo transfer(ET), the effective measure was explored in order to reduced ART multiple pregnancy rate. Methods:The data of 425 infertile couples were restrospectively analyzed,including pregnancy rate,pregnancy complications,delivery way and perinatal outcomes. Results:①There were IVF/ICSI-ET 4 603 cycles,the twin pregnancy rate was 23.35%,the abortion rate was 9.18%,the pregnancy complications occurrence rate was 13.41%,and the birth defects rate was 1.60%.②Between the two live born infant group and one live born infant group,there were statistically significant differences in fetal age[(29.66 ± 3.64) vs. (30.69 ± 3.47)week],birth weight[(2.64 ±0.45) vs. (3.12 ± 0.64)kg], cesarean delivery rate (97.68% vs. 77.38%),premature delivery rate (41.06% vs. 16.67%),low-weight baby occurrence rate(42.05% vs. 19.05)%. ③ Comparison of clinical data in mutifetal pregnancy reduction group and spontaneous reduction group with no reduction group,there were statistically significant differences in the fetal age[(38.00 ± 2.03),(37.89 ± 2.46) vs. (36.43 ± 1.77)week],birth weight [(3.05 ± 0.65),(3.15 ± 0.63) vs. (2.64 ± 0.45)kg] and premature delivery rate(13.64%,15.71% vs 37.24%). Conclusion:Compared the ART clinical reproduction outcome of twin with single live born infant,there were higher hazard in premature and lower birth weight. Women with ART pregnancies should get more perinatal care and antenatal examination. Clinical attention should be paid to diagnosis earlier,and multiple pregnancy reduction termination in time is very important for improving the perinatal outcomes.

Abstract:Objective:To analyze the molecular characteristic and antibiotic resistances of Vibrio cholerae O1 stains in Jiangsu province in 2010,and provide laboratory evidence for the clinical analysis and treatment of cholera. Methods:V. cholerae O1 strains in Jiangsu province in 2010 were analyzed by virulence genes,sequence analysis of ctxB gene,antibiotic resistances and PFGE. Results:All strains were positive for ctxA,ace,zot,toxR,tcpI,ompU,rtxC,tcpAEL,hlyAEL. They were also identified as harboring the classical ctxB genotype based on amino acid residue substitutions. They exhibited fully sensitivity to gentamicin,norfloxacin and ciprofloxacin,while they were all resistant to sulphamethoxazole/trimethoprim,streptomycin. The PFGE profiles of NotI showed a single banding pattern among 15 isolates,except VC201014. Conclusion:A single clone of a V. cholerae strain was responsible for the epidemic caused by V. cholerae O1 strains in Xuzhou,Huaian,Suqian,Nanjing during early September to late October in 2010,while there was another clone which was responsible for the epidemic in Lianyungang in late August in 2010. The bacterium type of V. cholerae causing the epidemics belonged to the atypical EL Tor variant. Gentamicin,norfloxacin and ciprofloxacin may be used for the treatment of this type of cholera.

Abstract:Objective:To identify the virus strain isolated from a suspected dengue case imported to Jiangsu province from Philippines in April 2012 and analyze its molecular characteristics. Methods:The serum sample collected from the patient was detected for IgM and IgG by colloidal gold method. The sample was further inoculated in C6/36 cells for virus isolation. Dengue nucleic acid was detected in the isolates via a stable cytopathic effect on C6/36 cells according to reverse translation-PCR and real-time PCR. Genome sequence information from the isolated strain was obtained by next-generation high-throughput sequencing. Nucleotide alignment and phylogenetic tree construction were conducted by MEGA6.0 software. Results:IgM and RNA of type 1 dengue virus were detected in the serum sample. The nucleotide homology with dengue virus 1 isolated from American Haw O3663 was more than 98%. Conclusion:The imported case of fever was caused by the dengue virus type 1.