Objective To explore the potential mechanism of severe liver injury shortly after withdrawal of antiviral therapy in chronic hepatitis B (CHB) patients.Methods Forty-nine patients with chronic hepatitis B virus (HBV) infection from the Department of Infectious Diseases of the First Affiliated Hospital of Nanchang University and 8 healthy volunteers from August 2014 to March 2015 were included in this study.All of them were human leukocyte antigen (HLA)-A2-positive.CHB patients were classified into three groups,including 15 cases in immune-tolerance group,20 cases in sustained antiviral treatment group,and 14 cases in recurrence of drug withdrawal group.The frequency of peripheral HLA-A0201-restricted hepatitis B core antigen (HBcAg)18-27 pentamer complex specific CD8+ T cells in CHB patients was analyzed by flow cytometry.Enzyme linked immunospot assay(ELISPOT) was used to detect interferon-gamma (IFN-γ) and tumor necrosis factor-α (TNF-α) secretions of HBcAg18-27-specific CD8+ T cells.The experimental data were analyzed using non-parametric U tests.Results In healthy control group,immune-tolerance group,sustained antiviral treatment group and recurrence of drug withdrawal group,the frequencies of HBcAg-specific CD8+T cells were (0.17 ± 0.16) ％,(1.46±0.72)％,(3.24± 1.60)％ and (4.67±2.43)％,respectively.Compared with healthy control group,the difference were all statistically significant in the three groups (Z=-3.583,-4.018 and-3.823,respectively;all P＜0.01).The frequencies of HBcAg-specific CD8+T cells in immune tolerance group or recurrence of drug withdrawal group were both significantly different from that in sustained antiviral therapy group (Z=-3.400 and-2.030,respectively;both P＜0.05).The difference between immune-tolerance group and recurrence of drug withdrawal group was also significant (Z =-3.230,P＜0.01).The secretion levels of IFN-γ of HBcAg-specific CD8+T cells in healthy control group,immune-tolerance group,sustained antiviral treatment group and recurrence of drug withdrawal group were2 (0-6),16 (2-53),106 (14-254) and 156 (28-395) spot forming cell (SFC)/106 peripheral blood mononuclear cell (PBMC),respectively.The differences between healthy control group and immune-tolerance group,sustained antiviral treatment group or recurrence of drug withdrawal group were all statistically significant (Z=-3.585,-4.069 and-3.824,respectively;all P＜0.01).The IFN-γ level of HBcAg-specific CD8+ T cells in recurrence of drug withdrawal group was significantly higher than that in sustained antiviral therapy group (Z=-2.205,P=0.027),and that in sustained antiviral therapy group was significantly higher than that in immune-tolerance group (Z=-4.700,P＜ 0.01).The TNF-α levels secreted by HBcAg-specific CD8+ T cells in each group were 2 (0-5),16 (2-32),112 (15-283),and 195 (55-537) SFC/106PBMC,respectively.The differences between healthy control group and immune-tolerance group,sustained antiviral treament group or recurrence of drug withdrawal group were all statistically significant (Z=-3.619,-4.069 and-3.824,respectively;all P＜0.01).The TNF-α level secreted by HBcAg-specific CD8+T cells in recurrence of drug withdrawal group was significantly higher than that in sustained antiviral therapy group (Z=-2.449,P=0.014),and that in sustained antiviral therapy group was significantly higher than that in immune-tolerance group (Z=-4.350,P＜0.01).Conclusions The changes of frequency and immune function of HBcAg-specific CD8+T cells in CHB patients may be one of the reasons causing severe liver damage after irregular withdrawal of nucleoside analogues.

Objectives To investigate the clinical features of progressive familial intrahepatic cholestasis type 2 (PFIC2) and to illustrate the importance of genetic diagnosis. Methods The mutations in 27 exons of ABCB11 encoding bile salt export pump (BSEP) were identiifed using polymerase chain reaction (PCR) and direct DNA sequencing in 6 children with suspected PFIC2. The pathogenicity of the newly identiifed mutations were predicted by SIFT, PolyPhen-2, SNPs&GO software. The clini-cal features and laboratory examinations were reviewed. Results Four disease-causing mutations, p.R928*, p.E554K, p.R575Q and p.Y337H were identiifed, and the last three mutations were novel. These three kinds of novel mutations can cause the disease. Two children with genetic diagnosis had such manifestations as onset within a month after birth, jaundice, hepatosplenomegaly, upset, increased levels of total bilirubin and direct bilirubin, GGT<100 U/L and high levels of total bile acid. Conclusions Genetic diagnosis is a potent tool for clinical diagnosis of PFIC2.

Objective To compare the clinical effects of the traditional operation and the new patch technique for the elderly women with pelvic organ prolapse.Methods 60 elderly women with pelvic organ prolapse patients were randomly divided into the traditional group and the modern group with 30 cases.The traditional group was treated with traditional surgery.The madern group was treated by new patch repair.Compare of the operation time,operative blood loss,catheter duration,hospital stay,postoperative recovery and 3 years recurrence rate in two groups.Results The catheter duration and hospital stay of the mederm group were significantly shorter than those of the traditional group (t =8.9471,14.2584,P ＜ 0.05).The postoperative recovery of the new group was better than the traditional group (x2 =5.9341,P ＜ 0.05).The 3 years recurrence rate of the mederm group was significantly lower than those of the traditional group (x2 =4.7059,P ＜ 0.05).The operation time of the mederm group was longer than those of the traditional group (t =4.4631,P ＜ 0.05) and the operative blood loss was more than the traditional group (t =2.0318,P ＜ 0.05).Conclusion The effect of the new patchin the treatment of elderly women with pelvic organ prolapse is better than the traditional surgery.

Objective To study the clinical significance of monitoring the urine iodine level in pregnant women.Methods The urine iodine levels were detected in 391 cases of pregnant women.The relativity was analyzed among the pregnancy gestation,pregnancy complications and the urine iodine levels.The corresponding health educa-tion of iodine nutritional knowledge was conducted according to the results of the determination.Results (1 )The urine iodine value was (124.73 ±70.61)μg/L in 391 pregnant women.The severe deficiency iodine rate,deficiency iodine rate and appropriate iodine rate of pregnant women were 37.85%,64.70% and 25.06%,respectively.(2) There was no significant difference in values of urine iodine among the pregnant women during the first,second and third trimester of pregnancy[(121.76 ±71.81)μg/L,(125.52 ±69.28)μg/L,(129.30 ±75.19)μg/L](t≤0.59, all P >0.05).The mean values were all lower than the appropriate values of WHO(150 ~249 μg/L).The low iodine rates were 67.20%,63.23% and 65.12%,respectively.There was no significant difference among them(χ2 =0.56, P >0.05 ).(3 )The urine iodine value of the pregnant women complicated with hyperthyroidism [(85.76 ± 53.09)μg/L]was significantly lower than that of primary hypothyroidism combined with pregnancy [(133.65 ± 73.41)μg/L],pregnancy combined with subclinical hypothyroidism[(136.50 ±79.19)μg/L]and no pregnancy complication[(119.34 ±66.43)μg/L](t≥2.76,P <0.01).The rate of iodine deficiency(87.88%)was highest (χ2 =8.72,P <0.05).(4)The urine iodine value after iodine nutrition intervention was significantly higher than those before in 77 pregnant women[(129.74 ±68.04)μg/L vs.(107.01 ±74.36)μg/L,t =1.98,P <0.05].The iodine deficiency rate dropped from 79.22% to 63.64%(χ2 =4.58,P <0.05).Conclusion In order to improve the appropriate iodine rate of pregnant women,it should be very important significance on monitoring the urine iodine level of pregnant women in particular with a complication of pregnancy and conducting iodine nutrition intervention in whole pregnancy gestation.

Objective To investigate the polymorphism of Brucella melitensis biovar 3 ( B.melitensis biovar 3) strains isolated from Guangdong province .Methods PCR assays followed by agar-ose gel electrophoresis and capillary electrophoresis based on the multiple locus variable number of tandem repeats (VNTR) analysis (MLVA) were performed to analyze 43 clinic isolates of B.melitensis biovar 3 strains isolated from clinical patients in Guangdong province .Results MLVA typing showed that the simi-larities of the analyzed locus among 43 strains of Brucella ranged from 68.2%to 100%.32 genotypes identi-fied among the isolates were identical (100%similarity).27 out of 43 strains (62.8%) were single geno-types, while the other 16 strains (37.2%) belonged to 5 other genotypes with 2 to 5 strains in each of them . Conclusion B.melitensis biovar 3 isolates showed polymorphism distribution in Guangdong province as in-dicated by MLVA typing analysis .Single-genotype isolates accounted for 62.8% of all studied strains.No predominant genotype was found among all isolates .

ObjectiveTo establish the urinary proteome profile of the metabolic syndrome ( MetS ) patients,compare the different urinary proteins between the MetS patients and the normal individuals,and analyze the function of the different proteins,so as to explore the pathogenesis of MetS.MethodsOvernight urine were collected from normal controls (n =6) and MetS patients ( n =6).Acetone precipitation method was used to precipitate proteins of urine.Intra-group proteins were mixed together,identified by reversed phase liquid chromatography-mass spectrometry/mass spectrometry and quantified relatively using spectral counting method.The functions of differential proteins were analyzed using Panther.ResultsA total of 807 and 630 proteins were identified respectively in normal controls and MetS patients.Comparing MetS patients with normal controls,sixty different proteins were found,of which 23 proteins were up-regulated and 37 proteins were down-regulated in MetS patients.In the up-regulated proteins,plasminogen was involved in the plasminogen activation cascade and isoform of alphaenolase,phosphoglycerate kinase 1 and fructose-bisphosphate aldolase B down-regulated in MetS patients were involved in the process of glycolysis and fructose metabolism.ConclusionsThe urinary proteome profile of patients with MetS was established by reversed phase liquid chromatography-mass spectrometry/mass spectrometry.Different proteins between MetS patients and normal people were identified.The plasminogen activation cascade,glycolysis and fructose metabolism play key roles in the pathogenesis of MetS.

Objective To investigate the etiological characteristics of Streptococcus pyogenes that caused scarlet fever in different periods in Guangdong province.Methods 22 isolates from different periods were analyzed through emm typing,PCR detection for super antigen genes,antibiotic susceptibility test and pulsed-field gel electrophoresis (PFGE).Results All isolates were susceptible to cefotaxim,levofloxacin and penicillin.Streptococcus pyogenes isolated after the year 2000 were 100％ resistant to erythrocin and clindamycin,but the resistant rate for strains isolated before the year 2000 was 9.1％ (1/11).There were 3 emm types indentified from 22 isolates including emm12.0 (59.09％,13/22),emm6.0 (36.36％,8/22) and emm1.0 (4.55％,1/22),which were detected in the isolates from the year 1997 and 2011,from 1978 and 1986,and from 2008,respectively.The positive rates for speA,speB,speC,speF,speG,speH,smeZ,and ssa genes detected by PCR were 54.55％,100％,100％,100％,100％,54.55％,0％,and 86.36％ respectively.Among all strains,95.45％ of the isolates carried 6 superantigen genes simultaneously.Three clusters of 10 PFGE subtypes were identified in 22 isolates.Cluster Ⅰ consisted of all strains from 1997 and one strain from 2011.Cluster Ⅱ consisted of strains isolated from 1978 and 1986.Cluster Ⅲ consisted of nine strains from 2011 and one from 2008.Conclusion S.pyrogenes isolates in Guangdong province were susceptible to penicillin but resistant to erythrocin.emm 12.0 accounted for the majority of the three types and there was a high frequency of super antigen genes.

Objective To study the molecular characters of porA and porB genes which encode outer membrane proteins (OMP), and predominated clonal complex of Neisseria meningitidis isolates from Guangdong province. Methods Eighteen Neisseria meningitidis isolates from Guangdong province during year 1967 to 2007 were recovered and reconfirmed by API NH biochemical system, and serogrouped by antiserum. The characters of porA and porB gene were analyzed by DNA sequencing. The allele profiles and the sequence types (ST) were determined by multilocus sequence typing (MLST). Based on their allelic profiles, the evolution relationship was analyzed by PHYLIP software. The predominant clonal complex was determined through comparing with the information of reference strains from the PubMLST database. Results For porA gene, type 20 was more frequently in the variable region (VR) 1 and type 9 in VR2 before year 2004. However, for porB gene, type 4 was more frequently in VR Ⅰ, type7 in VR Ⅳ, type 11 in VR Ⅴ,and type 10 in VR Ⅵ, respectively. The multi-types character was presented in VR Ⅴ and VR Ⅵ after2004. VR Ⅶ and VR Ⅷ can not be found among all the isolates except for one W135 isolate in 2007. Among the seven housekeeping genes, the polymorphism of abcZ was the lowest one with 4 allele numbers, while pgm was the highest one with 13 allele numbers. The predominant clonal complex was ST-5 before 2004. The ST-4821 clone complex appeared in 2004 and caused cases every year since then. More important, highly invasive ST-11 clonal complex firstly appeared in Guangdong in 2007. Conclusion The molecular characteristic of OMP genes presents polymorphism for the Neisseria meningitidis isolates from patients in Guangdong province during 1967 to 2007. ST-5 is the predominant clonal complex before 2004 and the highly invasive clonal complex is circulating in recent 3 years. It suggests that the surveillance based on laboratory should be further enhanced.

Objective To explore the preventive effect of Fluconazole on invasive fungous infections in premature infants.Methods Two hundred and thirty-four cases of preterm infants hospitalized in Jiangmen Central Hospital from Feb.2008 to Oct.2013 were analyzed by retrospective study.The subjects were divided into 3 groups based on whether Fluconazole was used and it was used orally or intravenously for the prevention of invasive fungous infections in premature infants.The Fluconazole prophylaxis was not given to group A,whereas oral Fluconazole prophylaxis [6 mg/(kg · times),every other day] and intravenous Fluconazole prophylaxis [3 mg/ (kg · times),once every 3 days] were applied in group B and group C.The incidence of invasive fungous infections and the clinical effects of Fluconazole on the 3 groups were evaluated and compared.Results The number of cases analyzed in the study were 53,115 and 66 cases in group A,group B and group C,respectively.There were no significant differences between the 3 groups in terms of gender,gestational age,birth weights,length of hospitalization,intravenous nutrition,and number of peripherally inserted central catheter,antibiotics and invasive mechanical ventilation use days (all P ＞ 0.05).The incidences of invasive fungal infections were 11.32％ (6/53 cases),6.96％ (8/115 cases) and 0 in group A,group B and group C,respectively.The prophylaxis effects between the 3 groups were significantly different (x2 =7.078,P =0.029).Group A and group B were not significantly different (x2 =0.905,P =0.342),but group C was better than group A and group B,and the differences were significant (P =0.007,0.028).Conclusions Prevention of invasive fungous infections by intravenous Fluconazole has good curative effects,and which can be used as the first choice.

Objective To evaluate effectiveness of application with FOCUS harmonic scalpel in papillary thyroid carcinoma (PTC) surgery.Methods 110 patients with PTC between June 2013 and December 2013 were divided into two groups randomly as harmonic scalpel group (group H) and conventional group (group C).Incision length,operative time,estimated blood loss,drainage volume within 24 h,in the second 24 h and in total after operation,the number of lymph node retrieval,post-operative complication and hospital stay were compared between two groups.Rusults There is significant different in factors of incision length [(5.5±0.4) cm in group H and (6.6±0.5) cm in group C (P ＜ 0.05)],operative time [(54.4±8.7) min in group H and (73.3±13.9) min in group C (P ＜ 0.05)],estimated blood loss [(14.2±6.0) ml in group H and (29.8±5.0) ml in group C (P ＜ 0.05)].There were no differences in drainage volume,the number of lymph node retrieval,post-operative complication and hospital stay between two group (P ＞ 0.05).Conclusion FOCUS harmonic scalpel could reduce incision length,operative time,estimated blood loss in PTC surgery.