Objective To analyze distribution and antibiotic resistance pathogenic bacteria in neonatal sepsis in 10 years. Methods The distribution of pathogens and their antibiotic resistance were retrospectively analyzed in neonatal sepsis from January, 2004 to December, 2013. The results were compared between 2004-2008 and 2009-2013. Results The percentage of Streptococcus agalactiae rose from 1.0%to 4.2%and fungi rose from 1.9%to 7.9%in all pathogens in past 10 years. But the distribution of pathogenic bacteria was not significantly different. The ESBLs of Escherichia coli were increased from 28.6%to 36.0%(P>0.05). The Escherichia coli resistant to imipenem and meropenem had not been found. The Staphylococcus resistance to oxacallin and ampicillin/sulbactam was increased. The Staphylococcus resistant to vancomycin had not been found. Conclusions The main pathogens of neonatal sepsis are coagulase-negative staphylococci and E. coli. The fungi and Streptococcus agalactiae infections are signiifcantly increased.

OBJECTIVE:To observe the short-term efficacy and ADR of tenofovir disoproxil fumarate(TDF)in the treatment of multi-drug resistant chronic hepatitis B(CHB). METHODS:32 patients with multi-drug resistant CHB were analyzed retrospec-tively,and HBV drug-resistant genes were detected before treatment;there were a number of points to resistance;they were gave TDF orally. The recovery rate of serum alanine aminotransferase(ALT),HBV-DNA conversion rate,lactic acid and renal function were observed before and after treatment. RESULTS:The recovery rate of ALT reached 100% at 3 months,and the conversion rate of HBV-DNA reached 96.88%. The lactic acid levels and renal dysfunction was not found during treatment. CONCLUSIONS:TDF take effect quickly on multi-drug resistant CHB,and no obvious ADR is found.

To determine CD4 + CD25 + T regulatory cells (Tregs),forkhead box P3 (FoxP3) mRNA expression and levels of cytokines secreted by peripheral blood mononuclear cells (PBMCs) in individuals responsive or unresponsive to hepatitis B (HB) vaccination,and to explore the relationships between immune response and immune regulatory cells or cytokines.Methods Based on the antibody against hepatitis B surface antigen (HBsAg) after HB vaccination,the CD4 + CD25 + Tregs frequencies in PBMCs from 18 responders,22 nonresponders and 10 non-immunized healthy controls were analyzed by flow cytometry.The expression of FoxP3 mRNA in PBMCs with or without stimulation of phytohemagglutinin ( PHA ) and HBsAg was analyzed by real-time quantitative PCR.Levels of IL-4,IL-12,IL-18,and IFN-γ secreted by PBMCs after PHA and HBsAg stimulation were analyzed by enzyme-linked immunosorbent assay.Results The ratio of CD4 + CD25 + Tregs to CD4 + T cells in the nonresponders was markedly higher than that in the responders (P ＜0.05),but lower than that in the controls (P ＜0.01 ).FoxP3 was differentially expressed among the responders,nonresponders,and controls in PBMCs before and after PHA and HBsAg stimulation,and nonresponders had the highest FoxP3 mRNA expression (P ＜ 0.05 or P ＜ 0.01 ).The content of IFN-γ by PBMCs after PHA and HBsAg stimulation was markedly lower in the nonresponders as compared with the controls and responders (P ＜ 0.05).However,there were no significant differences in the levels of IL-18,IL-4,and IL-12 from PBMCs after PHA and HBsAg stimulation between the responders and controls as well as the nonresponders (P ＞ 0.05 ).Conclusion CD4 + CD25 + FoxP3 + Tregs may be involved in the negative regulation of responses to hepatitis B vaccination.Immunologic non-responses to hepatitis B vaccination may be related to IFN-γhyposecretion in PBMCs.

Objective To observe the effect of laparoscopic surgery for patients with acute gallstone pancreatitis(AGP). Methods The clinical data of 134 patients suffered from AGP treated with laparoscopy surgery since 2000 in our hospital were analyzed retrospectively.Results All the patients were treated with laparoscopic surgery successfully. Among them, 21 cases were treated with laparoscopic cholecystectomy (LC);113 cases with LC and exploration of common bile duct,induding 75 cases received opening the pancreatic capsule and placement of irregation tubes for postoperative washing the abdominal cavity during the same operation. One hundred and tweent-six cases(94.0%) cured, 6 cases(4.5%) died, 2 cases( 1.5%) discharged themself. Conclusions Laparoscopic surgery in the treatment of early stage of AGP can get good results and improve the prognosis remarkably.It is worth to be used widely.

Objective To explore the value of application of choledochofiberscopy in the diagnosis and treatment of the extrahepatic bile duct disease, and the effect on reducing the incidence of the postoperative residual stone in biliary ducts. Methods According to the case history and ultrasonography,if the common bile duct(CBD) diseases suspected,the CBD was explored by intraoperative choledochofiberscope(IOCF). During the procedure,a biliary passage mirror inducer apparatus and biliary tract probe which were manufactured by ourselves were used. Results During LC,IOCF was performed on 385 cases of the 10 396 LC cases,and possitive findings were dicovered in 102 cases(26.49%). Among those positive patients, 67 cases belonged to stricture of the lower biliary tract; 5 cases were Mirizzi syndrome; 2 cases were carcinoma of the periampulla; 1 case was primarily carcinoma of the bile duct; 1case was ascarisis of the biliary system. Conclusions IOCF is a good inspect technique with high success rate and clear image of bile duct, it can discover the common duct diseases which are difficult to be diagnosed through the routine examination.At the same time, it can provide the locative and qualitive diagnosis, determine reasonable methods of operation,and effectively provent postoperative complications.

Objective To investigate the prognostic factors for clinical relapse in chronic hepatitis B (CHB) patients with hepatitis B e antigen (HBeAg) seroconversion after drug withdrawal and to establish a prognostic model.Methods Totally 201 CHB patients with HBeAg seroconversion after the antiviral therapy were enrolled.The epidemiological variables including age, gender and family history of hepatitis B were collected.Liver function and hepatitis B virus (HBV) DNA level one week before initiation of antiviral therapy, hepatitis B surface antigen (HBsAg) level at the time of drug withdrawal and the duration of antiviral therapy after HBeAg seroconversion were analyzed.The clinical relapse after 48 weeks of drug withdrawal was followed up.The patients were divided into relapse group and non-relapse group according to clinical variables at 48 weeks after drug withdrawal.The counting data were analyzed by chi-square test and the measurement data were analyzed by t test.The Logistic regression model was used to determine the prognostic factors for clinical relapse.The receiver operating charactenstic (ROC) curve was constructed to assess the performance of the prediction model.Results The clinical relapse rate was 16.42% (33/201) after 48 weeks of drug withdrawal.By multivariate analysis, age, the duration of antiviral therapy after HBeAg seroconversion and HBsAg level at the time of drug withdrawal were independent predictors (χ2=14.546, t=3.202, t=3.286, respectively;all P<0.05).The regression model Logit (P)=1.220×age-0.040×the duration of antiviral therapy after HBeAg seroconversion +0.004×HBsAg level at the time of drug withdrawal-5.426.The sensitivity and specificity with the cut-off value of-0.860 were 73.10% and 90.40%, respectively.Conclusions Age, the duration of antiviral therapy after HBeAg seroconversion and HBsAg level at the time of drug withdrawal are independent predictors for clinical relapse 48 weeks after drug withdrawal in CHB patients with HBeAg seroconversion after antiviral therapy.

Objective To discuss the clinical value of magnetic resonance diffusion-weighted imaging (MR-DWI) in distinguishing tumor remnants from tumor necrosis of pancreatic carcinoma after cryoablation treatment.Methods Conventional MRI T1WI,T2WI scan,DWI sequence and dynamic enhanced MRI scan were performed in 26 patients with pancreatic carcinoma who were received cryoablation treatment.The changes in MRI signals after cryoablation treatment were recorded.The apparent diffusion coefficient (ADC) values of the normal pancreas,preoperative tumor tissue,postoperative remnants and necrosis tissue were calculated,and the results were compared.The correlation between the ADC values and the size of the tumor was evaluated,and the differences in ADC values among the tumors that had different diameter,location and staging were statistically analyzed.Results Of the 26 patients,complete necrosis of tumor was obtained in 16.The necrotic tumor tissue displayed low-signal on T1WI,high-signal on T2WI and low-signal on DWI,with no enhancement on dynamic enhanced imaging.Active residual tumor tissue was detected in 9 patients,among them the residual tumor diameter ＞5 cm was seen in 7 patients;the residual rate was 34.6％.ADC values of the following tissue,from low to high in order,were preoperative pancreatic tumor tissue (1.022± 0.126)x10-3 mm2/s,postoperative residual tumor tissue (1.130±0.155)x10-3 mm2/s,normal pancreatic tissue (1.924±-0.124)×10-3 mm2/s and postoperative necrosis tissue (2.312-±0.214)×10-3 mm2/s.No statistically significant difference in ADC values existed between preoperative pancreatic tumor tissue and postoperative residual tumor tissue (P=0.452),while statistically significant difference in ADC values existed between normal pancreatic tissue and postoperative necrosis tissue (P＜0.001).The ADC values of pancreatic tumor tissue bore a negative correlation with the tumor size (R=-0.43,P=0.027 2),while the ADC values lacked the relationship to the tumor location as well as to the tumor staging (P=0.738 8 and P=0.089 5 respectively).Conclusion MR-DWI can effectively distinguish the residual tumor tissue from the necrotic tumor tissue of pancreatic carcinoma after cryoablation treatment,which provides reliable basis for further clinical diagnosis and treatment.

Objective To investigate the expression of specific marker molecules in hair-inducing activity of long-term cultured human dermal papilla cells (HDPCs) in vitro.Methods After dissected and cultured the HDPCs in vitro,the cells of passages 1 to 8 were used for experiments.The growth appearances of HDPCs in different passages were observed under inverted microscope.To detect the expression of specific marker molecules of long-term cultured HDPCs,the alkaline phosphatase (ALP) activity of the HDPCs was examined,and the specific genes ALP and insulin-like growth factor-1 (IGF-1) expression levels of HDPCs were determined by real-time quantitative PCR.Results After long-term cultured in vitro,the ALP and IGF-1 expression levels of HDPCs gradually decreased in different passages,as well as the display of the aggregated and cartouche growth.The ALP and IGF-1 expression levels of HDPCs in passage 1 was the highest,they were almost about 6.8-fold and 3.5-fold higher than the HDPCs in passage 8.The ALP staining of the HDPCs in passage 1 and passage 2 were evident,but the cells' ALP staining gradually became much weaker than the cells in the previous passages after the long-term cultured in vitro.Conclusions The expression levels of specific marker molecules ALP and IGF-1 of the HDPCs decrease gradually after long-term cultured in vitro,and the higher passage HDPCs lost the special aggregated and cartouche growth appearance,and hence lead to the loss of hair-inducing activity of HDPCs.

Objective To analyze the virulence genes and molecular typing of non-O1/non-O139 Vibio cholerae in bloodstream infection,and to provide the scientific basis for its diagnosis,treatment, prevention and controls.Methods Five Vibio cholerae strains were obtained from blood samples of five inpatients with sepsis in Ruian People ’s Hospital from 2012 to 2015 . Morphological examination, biochemical identification,drug sensitivity test and multilocus sequence typing (MLST)classification analysis of strains were conducted.Totally 17 virulence genes were detected by PCR amplification.Results These five suspected Vibrio cholerae isolates were confirmed as non-O1/non-O139 Vibrio cholerae . Drug susceptibility test showed that all the strains were sensitive to tetracycline,ciprofloxacin,piperacillin and tazobactam, meropenem, amikacin and gentamicin; one strain was resistant to trimethoprim/sulfamethoxazole;all were resistant to ampicillin.MLST analysis showed that all strains were new sequence types (ST),belonging to ST268,ST269,ST267,ST270 and ST271 ,and two novel alleles of RY03(mdh:60 and pyrC:86)were discovered.Virulence genes testing showed that the five strains were divided into 4 virulence genotypes:RY02 and RY04 (hlyA + toxR + hap + rtxA + nanH + vasH + vasA +vasK + ),RY01 (hlyA +toxR +hap +rtxA +nanH +vasH -vasA +vasK - ),RY03 (hlyA +toxR +hap +rtxA +nanH - vasH + vasA + vasK + ) and RY05 (hlyA + toxR + hap + rtxA + nanH + vasH - vasA - vasK - ). Conclusions Non-O1/non-O139 Vibrio cholerae can cause human bloodstream infection in immunocompromised patients.The pathogenic factors may be related to the virulence genes of hlyA, toxR,hap ,rtxA and T6SS.

Eight patients with suspected cases of C .jejuni were etiologically diagnosed and analyzed in this study to pro-vide scientific basis for the confirmation of the cases of human campylobacteriosis in Guizhou Province ,China .Blood or feces of 8 suspected patients were employed to isolate bacteria strains .Conventional and multi-PCR techniques were applied to identify suspicious bacteria strains .The C .jejuni strains were analyzed by using Pulsed Field Gel Electrophoresis (PFGE) .Suspicious strains of C .jejuni were isolated from all the 8 suspected patients of campylobacteriosis and anticipated genes fragment were detected with multi-PCR .With the digestion of restriction enzyme SmaI ,the 8 C .jejuni strains were divided into 7 PFGE pat-terns with 7-10 DNA bands .Cluster analysis showed that the gross similarity of 8 strains of C . jejuni was more than 50% . The similarity of PFGE patterns between strain GZ201004 and GZ201005 from diarrhea patients was as high as 100% ,while the similarity of strain GZ201201 and GZ201007 was 66 .7% .Moreover ,C . jejuni were detected from all the suspected pa-tients of campylobacteriosis .PFGE results indicated that strains GZ201004 and GZ201005 were from the same source ,while all the 8 isolates showed PFGE polymorphism .