Objective To explore the expression of interferon(IFN)-γ,interleukin(IL)-4 and IL-17 in the patients with hepatitis B virus(HBV)infection,which can reflect the function of helDer T lymphocyte(Th)1,Th2 and Th17 cells.Methods There were 34 chronic hepatitis B(CHB)Datients,31 liver cirrhosis(LC)patients,26 primary hepatocellular carcinoma(PHC)patients,22 chronic severe hepatitis patients and 15 healthy controls involved in our research.The levels of IFN-γ,IL-4 and IL-17 in serum of the patients with chronic HBV infection and healthy controls were determined by enzyme linked immunosorbent assays(ELISA). And the dala were analysed via t-test and Pearson correlation analysis.Results IFN-γ/IL-4 ratio of the CHB group(1.08±0.66,P＜0.01)was lower than that of healthy controls(2.60±0.60),and the ratio in the chronic severe hepatitis B group(4.81±0.87,P＜0.01)was higher than that in healthy control group.IFN-γIL-17 ratio in the CHB(1.13±0.85,P＜0.01),the LC(1.69±0.92,P=0.010)and the PHC group(1.76±0.84,P=0.011)were all lower than that in healthy conlrols group(2.66±0.70),and the ratio in the chronic severe hepatitis B group(3.68±0.42)was higher than that in healthy controls group (P=0.004).IL-4/IL-17 ratio in the LC(0.72±0.38,P=0.026)and the PHC group(0.63±0.19,P＜0.01)were both lower than that in heahhy controls(1.04±0.23).Conclusiolls The dominant expression of IL-4,IL-7 and IFN-γ is respectively found in the CHB patients,LC and PHC patients,and chronic severe hepatilis B patients.

Objective To compare the efficacy and safety of vitrectomy combined with internal limiting membrane flap and vitrectomy combined with internal limiting membrane peeling for the treatment of idiopathic macular hole with different sizes.Methods A total of 127 consective patients (127 eyes)were divided into two groups according to the size of the hole diameter of the smallest split points by less than or equal to 500 μm (small diameter macular hole group) and more than 500 μm (huge diameter macular hole group).According to different surgical methods the patients were divided into non ILM flap coverage group (peeling 1 group and peeling 2 group) and ILM flap cover group (covering 1 group and covering 2 group).All the patients underwent vitrectomy combined with internal limiting membrane peeling or vitrectomy combined with limiting membrane flap.Preoperative and postoperative best correct visual acuity,closure ratio of macular hole and postoperative major complications were observed and followed up.Results The postoperative best correct visual acuity improved in all the groups,there was no significance difference between small diameter macular hole group and huge diameter macuiar hole group (t =0.112 2,0.750 8;all P ＞0.05).The closure ratio of peeling 1 group and covering 2 group at postoperative 6 months were all 100％,there was no statistical difference (P ＞ 0.05),which in peeling 2 group and covering 2 group were 84.85％ and 100.00％,there was statistical difference (x2 =13.292,P ＜ 0.05).There was no statistical difference in preoperative defect diameter of the inner and outer junction between peeling 2 group and covering 2 groups (P ＞0.05),there was also no statistical difference between peeling 2 group and covering 2 groups at postoperative 1 months (P ＞ 0.05),but there were statistical differences at postoperative 3 months,6 months and 12 months (all P ＜ 0.05),the covering 2 group were less than the peeling 2 group.Conclusion ILM flap coverage helps to heal macular holes greater than 500 μm diameter,and has no extra effect on healing of diameter less than 500 μm.

Abstract: The activities of four CYP450 enzymes (CYP3A, 1A2, 2El and 2C) and the mRNA expression levels of CYP1A2, 2El, 2Cll and 3A1 in rat liver were determined after Wistar rats were orally administered with brucine (BR) at three dosage levels (3, 15 and 60 mg.kg-1 per day) and the high dose of BR combined with glycyrrhetinic acid (GA, 25 mg.kg-1 per day) or liquiritin (LQ, 20 mg.kg-1 per day) for 7 consecutive days. Compared with the control, brucine caused 24.5% and 34.6% decrease of CYP3A-associated testosterone 6beta-hydroxylation (6betaTesto-OH) and CYP2C-associated tolbutamide hydroxylation (Tol-OH), respectively, and 146.1% increase of CYP2El-associated para-nitrophenol hydroxylation (PNP-OH) at the high dose level. On the other hand, (BR+GA) caused 51.4% and 33.5% decrease, respectively, of CYP2El-associated PNP-OH and CYP1A2-associated ethoxyresorufin-O-de-ethylation (EROD) as compared with the high dose of BR group. Meanwhile, (BR+LQ) caused 41.1% decrease of CYP2El-associated PNP-OH and 37.7% increase of CYP2C-associated Tol-OH. The results indicated that the co-administration of BR with GA or LQ had effect on mRNA expression and activities of the CYP450 enzymes mentioned above to some extent, and the in vivo antagonism of LQ on BR-induced CYPs adverse effects and the in vivo inhibitory action of GA on CYP2E1 and 1A2 might play an important role in the detoxification of Radix Glycyrrhizae against Strychnos nux-vomica L.

Objective To explore the characteristics and surgical managements of recurrent retinal detachment in silicone oil tamponade eyes.Methods The records of consecutive series of 134 patients (134 eyes) with recurrent retinal detachment in silicone oil tamponade eyes from January 2012 to December 2015 in our hospital were reviewed retrospectively,the vitrectomy combined with silicone oil replacement or scleral buckling procedure were performed.The follow-up time was 6 months,the surgical efficient was evaluated.Results 101 eyes underwent vitrectomy combined with silicone oil replacement.Retina was completely reattached in 79 eyes,and vitrectomy was reperformed in the left 22 eyes,the successful rate was 78.2％ (79/101);The visual acuity improved in 16 eyes,unchanged in 56 eyes,decreased in 29 eyes;The intraocular pressure of 31 eyes increased more than 25 mmHg (1 kPa =7.5 mmHg).33 eyes underwent scleral buckling procedure.Retina was completely reattached in 23 eyes,and vitrectomy combined with silicone oil replacement was performed in the left l0 eyes,the successful rate was 69.7％ (23/33);The visual acuity improved in 5 eyes,unchanged in 16 eyes,decreased in 12 eyes;The intraocular pressure of 14 eyes increased more than 25 mmHg.Conelusion For limited retinal detachment caused by inferior or peripheral holes,proliferative vitreoretinopathy in the A or B-class,the refractive medium does not affect the fundus examination,scleral buckling surgery is preferred;For the hole in the posterior pole or extensive retinal detachment caused by giant retinal holes,proliferation or retinal fixed fold formation,vitrectomy combined with silicone oil replacement is a better option.

Objective To screen B and T cell antigen epitopes on Acinetobacter baumannii outer membrane protein 33×103-36×103 (OMP33-36).Methods B and T cell epitopes on OMP33-36 of Acinetobacter baumannii were predicted by bioinformatics methods and synthesized.Recombinant expression plasmid pET-30a-OMP33-36 was cloned and used to express OMP33-36 in a prokaryotic expression system.The expressed OMP33-36 was used to immunize BALB/c mice after purification.Serum sample was collected from each mouse in immunization and negative control groups, and then analyzed by indirect ELISA with synthesized peptides to identify B cell epitopes.Splenocytes were separated from every mouse and then cultured with each of the synthesized peptides, respectively.Double sandwich ELISA was performed to detect IFN-γ secretion in the supernatant of cell cultures for screening of T cell epitopes.Results Candidates of B and T cell epitopes were constructed, which were PB1, PB2, PB3, PT1, PT2 and PT3.Results of the indirect ELISA showed that peptides PB1 and PB2 reacted with the serum samples collected from immunized mice and A450 values of the immunization group were significant higher than those of the negative control group.Compared with the negative control group, enhanced secretion of IFN-γ following peptide PT3 stimulation was observed in the immunization group as indicated by the double sandwich ELISA.Conclusion Two B cell epitopes PB1 and PB2, and one T cell epitope PT3 on the OMP33-36 of Acinetobacter baumannii were successfully constructed and screened out.

0.05).There was significant difference in the optical density of the 43kDa protein between the patients with OAB and controls(P

Objective To observe the activation of anti-viral innate immune response of type Ⅰinterferon and inhibition of hepatitis B virus (HBV)genome replication in mice by HBV-3p-siRNA. Methods HBV-3p-siRNA was designed by targeting specific sequence of HBV S/P mRNA and was generated by in vitro transcription.Negative control siRNA (NC-siRNA)and non-modified HBV-siRNA were used as control groups.Blood samples were collected from tail vein of mice and the model of HBV-infected mice were established by hydrodynamic injection.Forty mice were divided into 4 groups with 10 in each group.The model group was only injected with pGL3.0-HBV1 .2 copy plasmid.The negative control group received peritoneal injection of NC-siRNA.HBV-siRNA group received peritoneal injection of HBV-siRNA and HBV-3p-siRNA group received peritoneal injection of HBV-3p-siRNA.The interferon-β(IFN-β)and hepatitis B surface antigen (HBsAg)in serum were detected by enzyme linked immunosorbent assay (ELISA).The copies of HBV DNA were assessed by fluore scence quantitative polymerase chain reaction (PCR ).The statistical difference between groups was determined using One way-ANOVA analysis by LSD or Dunnett T3.Results Serum level of IFN-β was (12.37±5 .32)pg/mL in model group,(22.61 ±6.29 )pg/mL in negative control group,(26.40±5 .39)pg/mL in HBV-siRNA group and (68.37± 21 .00 ) pg/mL in HBV-3p-siRNA group.The secretions of IFN-β into serum were significantly enhanced by HBV-siRNA and HBV-3p-siRNA compared with model group (F =23.988 and 46.523,respectively,both P <0.01).Serum level of HBsAg was (2 864.86±907.11 )ng/mL in model group,(2 198.86±456.89 )ng/mL in negative control group,(1 049.71 ± 396.28 )ng/mL in HBV-siRNA group and (640.86±383.08)ng/mL in HBV-3p-siRNA group.The expressions of HBsAg were inhibited by HBV-3p-siRNA and HBV-siRNA compared with model group (F = 23.537 and 39.144, respectively;P =0.025 and 0.010,respectively).Serum level of HBV DNA was (2.54 ×104 ±1 .46 × 104 )copy/mL in model group,(2.22×104 ±2.62×103 )copy/mL in negative control group,(3.59×103 ±2.88×103 )copy/mL in HBV-siRNA group and (2.65 ×103 ±1 .46×103 )copy/mL in HBV-3p-siRNA group.Serum level of HBV DNA were inhibited by HBV-3p-siRNA and HBV-siRNA compared with model group (F =15 .013 and 16.741 ,respectively,both P <0.05 ).All of the indicated siRNA used in the experiments showed no apparent effects on the body mass index of the mice models.Conclusion HBV-3p-siRNA,which induces the production of IFN-β and inhibits HBV replication through gene silencing in vivo ,may be a powerful bifunctional antiviral molecule.

Objective To characterize the antibiotic resistance,homology and carbapenemase genotypes of imipenem resistant Acinetobac1ter baumannii isolated from our hospital,and analyze the clonal relatedness of the test strains.Methods Ninety five strains of imipenem resistant A.baumannii were isolated from August 2003 to December 2004 in the First Affiliated Hospital, College of Medicine,Zhejiang University.The MICs of 16 antimicrobial agents against these strains were determined by agar dilution and E-test method.The homology of these isolates was analyzed by pulse-field gel electrophoresis(PFGE).The coding gene of carbapenemases was amplified.PCR products were purified,cloned and sequenced.Plasmid DNA was extracted and purified.Conjugation and Southern blot were performed to locate the position of oxa 23 gene.Results The resistance rates to ampicillin-sulbactam and cefoperazone sulhactam were 67.9% and 30.2%.Polymyxin E had the lowest resistance rate of 17%. The resistance rate to other antimicrobial agents was higher than 90%.The 95 strains,isolated from 10 clinical units,were classified into 6 clones.Clones A and B were predominant clones.All strains produced carbapenemases which were confirmed as OXA 23 by PCR and sequencing analysis.No plasmid was extracted and conjugation was not successful.Southern bolt showed that oxa-23 gene was located on Apal-digested chromosomal segments about 220 kb and 200 kb in Clones A and B,re spectively.Conclusions OXA 23-producing A.baumannii has become one of the most important multi-resistant pathogens in our hospital.Clones A and B have widely spread in our hospital.Oxa-23 gene is located on chromosomal DNA.

Objective To investigate the resistant mechanism of imipenem-resistant K. pneumoniae.Methods The minimal inhibitive concentrations (MICs) of the antimicrobial agents were determined by Etest.Isoelectric focusing electrophoresis (IEF),plasmid extraction,conjugation, transformation,PCR amplification,cloning and sequencing were carried out for analyzing the encoding gene of ?-1actamases.Results Three kinds of ?-1actamases were detected with pIs of 7.2,6.7,and 5.4.in a clinical strain of K.pneumoniae.These ?-1actamases were TEM-I (pI,5.4),SHV-12 (pI,8.2) and KPC-2 ( pI,6.7 ) confirmed by sequencing of the PCR products.Only one band of ?-1actamase with pI 6.7 was displayed in the transformant.A 1500 bp segment,which contained the KPC-2 gene confirmed by nucleotide sequence analysis,was cloned from a 60 000 bp plasmid of the transformant.Conclusion The strain of K.pneumoniae resistant to imipenem produces a plasmid-mediated carbapenemase KPC-2 which belongs to Bush group 2f,class A ?-1actamase.

Objective To compare the infectivity between laboratory adapted human inununodefi- ciency virus(HIV-1) and primary HIV-1 isolates for different mucosal epithelial cell lines. Methods Mu-cosal epithelial cells Caco-2, T-84, HeLa and lymphocyte MT-4 were infected with laboratory adapted HIV-1 SF33 and 2 primary HIV-1 isolates (02010561, 02010141). Culture supernatant and cells were collected respectively on 3-4 days interval after virus inoculation. The former was tested for HIV-1 antigen P24 level and viral load, and the latter was tested for total viral DNA and integrated viral DNA. Results All 3 virus strains could infect MT-4 cells and integrate into their genome. Only HIV-1 SF33 could infect Caco-2 cells but could not integrate into their genomic DNA. Both HIV-1 SF33 and 02010561 infected HeLa cells but only integration of HIV-1 SF33 was detected. All the 3 HIV-1 strains infected T-84 cells but only the integra-tion of HIV-1 SF33 and 02010141 was observed. Conclusion Although laboratory adapted and primary HIV-1 strains are able to infect human mucosal epithelial cell lines, transient or productive infection estab-lished in different mucosal epithelial cells is dependent on the character of cells and virus strains.