Granulysin is a cytotoxic granule protein,colocalizes with perforin and granzymes in the cytolytic granules of human CTL and NK cells. The released granulysin was found in the sera of healthy individuals and patients. Granulysin exhibits potent cytotoxic activity against tumors and a broad panel of microbes, including bacteria, fungi, and parasites. The serum levels of granulysin is well associated with diverse activities of NK cells and CTL in physiological and pathological settings and could be a useful novel serum marker for acute rejection of grafts and the overall status of host cellular immunity.

Objective To analyze the characteristics and hepatotropism of this negative element HBVnt453-250 sequence.Methods pHBv453-250,pHBV250-453.plucHBV453-250 and plucHBV250-453 were constructed,with luciferase and enhanced green fluorecence protein(EGFP)gene as the reporter gene,respectively.After transfection of HepG2 cells with these plasmids,luciferase assays,real-time PCR and western blot assays were used to detect the gene transcription and expression level.The SV40 promoter of pGL3 control and pHBV453-250 were replaced by the cytomegalovirus early promoter,resulting in plasmids pCMVcontrol(luc)and pCMV453-250(luc).Results Compared with pHBV453-250,the mutant plasmids.with the inhibitory element inserted in difierent site or inverted orientation.exerted similar downregulation of Juciferase gene transcription and expression.Western blot analysis demonstrated the similar repression when EGFP was used as the reporter gene.By transfeeted to HepG2 cell line,the plasmid pCMV453-250(1UC)could reduce lneiferase activity(36.56%)compared with pCMLcontrol(luc).When the plasmids plueHBV453-250 and plucHBV250-453 were transfected to non-liver cell lines(A549,HeLa),luciferase gene was expressed weakly,compared with that of pGL3control(P＜0.05).Conclusion The inhibitory effect of HBVnt453-250 sequence acted in both orientation-and position-independent manners,and had no promoter selectivity and funotioned in hepatocyte-independent manner.

Objective To investigate the diagnostic and prognostic value of presepsin for sepsis.Methods Diagnostic accuracy test.The plasma presepsin levels of 57 sepsis patients,64 systemic inflammatory response syndrome (SIRS) patients and 120 healthy individuals admitted to the 263 Clinical Branch,General Hospital of Beijing Military Region between January 2012 and December 2013were detected by PATHFAST system.Receiver operating characteristic (ROC) curve analysis was used to assess and compare the diagnostic value of presepsin and procalitonin (PCT).Logistic regression model was used to estimate the association between presepsin and sepsis.In addition,the correlations between presepsin and the clinical characteristics were analyzed in sepsis patients.Results Sepsis patients [1 266 (754-2 181) pg/ml] had higher presepsin level than SIRS[517 (349-939)pg/ml] and healthy individual controls [(182 ± 56)pg/ml] (Z value was 5.94 and 10.71,respectively,P value was ＜ 0.01 for all).The areas under curve (AUCs) of presepsin and PCT were 0.81 (95％ CI:0.74-0.89) and 0.78 (95％ CI:0.71-0.86),respectively,with no statistical significance (x2 =0.60,P =0.47).After adjusted for PCT,presepsin ＞ 1 060 pg/ml was independently associated with sepsis,with odds ratio (OR) of 7.80 (95 ％ CI:3.07-20.32).Severe sepsis patients [2 723 (2 002-4 234) pg/ml] had higher presepsin than sepsis patients[1 145 (656-1 436) pg/ml] (Z =4.00,P ＜0.01).The patients with inhosptal mortality [2 365 (1 256-3 567)pg/ml] had higher presepsin than survival ones[1 146 (660-1 452) pg/ml] (Z =2.99,P =0.003).Presepsin was positively correlated with PCT (r =0.75,P ＜ 0.01).The reference for presepsin was 72 to 292 pg/ml.Conclusions Presepsin was an useful biomarker for sepsisdiagnosis.The diagnostic value of presepsin and PCT was not completely overlap,and combinational using of these two biomarkers may improve the diagnostic accuracy of sepsis.In addition,presepsin had potential value for prognosis estimation.

Objective To study the application effect of fibrin sealant(FS) during operation for late-stage portal hypertension caused by schistosomiasis.Methods From Jun,2003 to Jun,2005,92 cases of(late-stage) portal hypertension caused by schistosomiasis treated by portal-systemic disconn-ection(PSD)(operation) were divided into two groups,namely PSD group and PSD+FS group.The early complications such as fever and exudate in splenic fossa,as well as long term complications such as recurrent bleeding,(hypertensive) gastropathy were compared in the 2 groups,and encephalopathy were compared between 64 cases of patients with FS and 28 cases of patients without FS.Results The patients undergoing PSD+FS showed decreased the exudate in splenic fossa and fever(P0 05).Conclusions FS application during operation is effective in attenuating early and long term operative complications of portal hypertension caused by(schistosomiasis),and improving outcome of surgical treatment.

Objective To detect the recombinant intermediates of hepatitis B virus (HBV) between genotype B and C in vitro. Methods Vector Plenti6/V5-D-topo-X was genetically modified by genotype B and C to transfect HepG2 cells. Then the HepG2 cells were amplified and sequence of the nucleic acid after the transinfection was tested and compared with RDP3Beta40 software package and bootscanning procedure in SimPlot program package. Results Three recombinant intermediates of HBV between genotype B and C were identified in vitro. Genotype C in the precore region plus the core gene spanning nucleotide positions from 1740-1838 to 2443-2485 contributed to the recombination with genotype B. Isolate R1 recombinant intermediate had 2 break points at nt2170-2172 and nt2188-2189. Nucleic acid changed from CAC to TGT and from GA to AC, respectively. Isolate R2 recombinant intermediate had a break point at nt1740-1 838, and 3 bases changed in different nucleic acid sites: from A to T at nt1740, from C to T at nt1753, and from G to A at nt1838, respectively. Isolate R3 recombinant intermediate had a break point at nt2443-2483, and 4 bases changed in different nucleic acid sites: from C to T at nt2443, from A to G at nt2452, from T to C at nt2480, and from C to T at nt2483, respectively. Conclusion The recombinant intermediates of HBV between genotype B and C have been detected in vitro and the changes have been identified in the precore region plus the core gene in genotype B and C.

Direct-acting antivirals (DAAs) play a critical role for the therapy of chronical hepatitis B. DAAs can decrease the production of viral progeny of hepatitis B virus (HBV), breaking the viral dynamic equilibrium between: (1) virion production from hepatocytes and clearance from circulation; (2) replenishment and decay of covalently closed circular (ccc)DNA pool inside infected hepatocytes. Nucleos(t)ide analogues can potently shift the first balance to undetectable viremia in the blood, but have limited or no effect on the second one, thus making it imperative to develop new agents targeting additional step(s) of HBV life cycle. We herein briefly introduce the DAAs currently in development by classifying them as agents affecting the replenishment or the decay of cccDNA pool.