Objective To identify human leucocyte antigen (HLA)-A* 0201-restricted hepatitis C virus (HCV)-cytotoxic T lymphocyte (CTL) epitopes. Methods Based on the prediction results of RANKpep and SYFPEITHI prediction programs, six candidate CTL epitopes were selected and synthesized. The affinity of candidate CTL epitopes to HLA-A* 0201 molecules of T2 cells was explored. Subsequently, enzyme-linked immunosorbent spot (ELISPOT) assay and intracellular cytokine staining (ICS) were utilized to determine whether candidate CTL epitopes could induce the recall positive response in peripheral blood mononuclear cells (PBMC) of HLA-A* 0201 positive HCV-1b-infected patients. Results Among six candidate CTL epitopes, peptides C_181(LLSCLTTPV) and NS2_172 (VLQAGLIRV) had high affinity to HLA-A* 0201 molecules. Moreover, the affinity was proportional to the concentration of peptide. Furthermore, among ten HLA-A* 0201 positive HCV-1b-infected patients, the frequencies of C_181 and NS2_172-specific interferon (IFN)-γ-producing cells were 0-19 spots forming cells (SFC)/1 × 105 PBMC and 0-20 SFC/1 × 105 PBMC, respectively.The percentages of C_ 181 and NS2_172-specific IFN-γ+ CD8+ T lymphocytes in total CD8+ T lymphocytes were 0.006%-0.065% and 0.005%-0.080%, respectively. Conclusion Peptides C_181 (LLSCLTTPV) and NS2_172 (VLQAGLIRV) are identified as novel HLA-A* 0201-restricted HCV-CTL epitopes.

Objective To investigate the level of chemotactic factors(CXCL5 and CXCL8)in hepatic fibrosis and cirrhosis tissue.Methods Hepatic tissues were obtained from 9 patients with hepatic hemangioma (hepatic hemangioma group),10 patients with liver fibrosis(liver fibrosis group)and 11 patients with liver cirrhosis(1iver cirrhosis group)at Nanfang Hospital from May 2008 to May 2009.The contents of CXCL5 and CXCL8 in hepatic tissue were assayed by ELISA.All data were analyzed by one-way ANOVA,Pearson rank correlation or Spearman correlation.Results The contents of CXCL5 and CXCL8 were(0.8±0.7)ng/g and(6.2±3.7)ng/g in hepatic hemangioma group,(2.0±2.0)ng/g and(11.6±3.5)ng/g in liver fibrosis group and (17.1±4.8)ng/g and(12.3±3.9)ng/g in liver cirrhosis group,with significant difference among the 3 groups (F=60.050,7.690,P＜0.05).The expression of CXCL5 was correlated with the content of alanine aminotransferase(ALT),aspartate aminotransferase(AST)and prothrombin time(PT)(r=0.502,0.468,0.523,P＜0.05):the expression of CXCL8 was correlated with the content of ALT,AST.total bilirubin and PT(r=0.477,0.504,0.537,0.431,P＜0.05).Conclusions With the aggravation of hepatic fibrosis,the contents of CXCL5 and CXCL8 are increased with different patterns.The changes of CXCL5 and CXCL8 are related with the injury of liver,but the changes of CXCL5 and CXCL8 do not correspond with the degree of the injury of liver.

Objective To construct and identify an eukaryotic expression plasmid containing rat hepatocyte growth factor(rHGF)gene and rat augmenter of liver regeneration(rALR)gene,so that to provide experimental basis for developing new treatments of hepatic fibrosis.Methods The gene fragments of rHGF and rALR were amplified from recombinant prokaryotic plasmid pUC18-rHGF and pUC18-rALR by polymerase chain reaction(PCR),respectively,then were spliced by overlap extension PCR with a linker,and the fusion gene rHGF-linker-rALR was constructed.The fusion gene was directionally inserted into the eukaryotic expression plasmid pcDNA3.1 between restriction sites of Kpn Ⅰ and Xba Ⅰ to construct the recombinant eukaryotic expression plasmid pcDNA3.1-rHGF-linker-rALR,and the new constructed recombinant plasmid was identified by double restriction digestion and DNA sequencing.Results DNA fragments of 2200 bp and 400 bp were observed after the electrophoresis of products amplified from recombinant prokaryotic plasmid pUC18-rHGF and pUC18-rALR,respectively,which was consistent with the theoretical value.The electrophoresis of fusion gene rHGF-linker-rALR obtained by overlap extension PCR technique showed only a 2 600 bp DNA fragment,which was in accordance with the expected value.Electrophoresis of products of pcDNA3.1-rHGF-linker-rALR digested with Kpn Ⅰ and Xba Ⅰ showed two DNA fragments with 2600 bp and 5400 bp,which were both consistent with the expected value.The sequences were confirmed correctly by DNA sequencing.Conclusion The recombinant eukaryotic expression plasmid pcDNA3.1-rHGF-linker-rALR is successfully constructed,which provides experimental basis for developing gene therapy of hepatic fibrosis.

To confirm the etiology of a dead case for a 6 year-old female Ailurus fulgens,one strain of the predominant bacteria from pathologic tissues(heart,liver,spleen,lung and other samples) of the dead Ailurus fulgens were examined and isolated.The isolate was named R1 and no other bacteria were isolated.The bacterial etiological examination(morphological characteristics,biochemical characteristics and 16S rDNA gene detection)of R1 showed that it was identifed as K.peneumoniae.Artificial infection to mice about R1 was also conducted in this study.R1 had strong pathogenicity to mice and the LD50 is 6.5 × 104 CFU/mL.Moreover,the clinical and pathological features of the dead mice were consistent with that of the Ailurus fulgens.To find effective therapeutic drugs of curing other Ailurus fulgens,antibiotic sensitivity test of R1 was conducted,and the results revealed that R1 was highly sensitive to cefotaxime et al,moderately sensitive to amikacin and resistant to penicillin.These data showed that K.peneumoniae was bacterial pathogen leading to death of the Ailurus fulgens and it had strong resistance to penicillins,macrolides and virginiamycin and it had broad drug resistance spectrum.However,R1 is sensitive to cephalosporins and aminoglycoside antibiotics.

Objective Previous studies have shown that vitamin D deficiency is closely related to cardiac remodeling. How?ever, the underlying mechanisms have not been fully elucidated. Moreover, oxidative stress plays an important role on the pathologies of cardiac remodeling. The aim of this study was to explore the influence of VD deficiency on cardiac oxidative stress and the potential sig?nal pathway. Methods The male C57 mice ( 3 weeks old) were randomly divided into three groups: vitamin D deficiency ( VDD ) group ( vitamin D deficiency feed for 10 weeks) , vitamin D deficiency ( VDA) group ( vitamin D sufficiency feed for 10 weeks) and VDD+calcitriol ( CAL) group ( vitamin D deficiency feed for 10 weeks and then vitamin D sufficiency feed and calcitriol treatment for 10 weeks) . Results There were significant differences between the VDD group and the VDA group in the left ventricular end?diastolic diameter and left ventricular mass index (3.82±0.125 mm vs 3.748±0.092 mm, P<0.05) (119.30±8.54 vs 97.60±3.65, P<0.05). The number of myocardial cells stained with 8?OHDG was higher in the VDD group compared with the VDA group ( 65. 4 ± 2. 3 vs 21. 8 ± 1. 6, P<0.05) whereas was lower after calcitriol supplement. Furthermore, the expression of thioredoxin interacting protein (TXNIP) was sig?nificantly up?regulated and the ratio of p?ASK?1/ASK?1, cytochrome C release, and caspase3 activation were increased in the VDD group . Conclusion VDD can lead to cardiac oxidative injury and the up?regulation of TXNIP and the activation of ASK?1 related apoptotic signal cascade may be involved in this procedure.

AIM: To study the effect of mesenchymal stem cells (MSCs) infusion on hematopoietic recovery after peripheral blood stem cell transplantation in mice. METHODS: BALB/c mice conditioned by high dose chemotherapy/radiotherapy were infused with 10 6 peripheral blood mononuclear cells (PBMC) mobilized by granulocyte colony-stimulating factor (PBSCT group), 10 4 MSCs culture-expanded in vitro and 10 6 PBMC(experimental group 1), 10 6 MSCs and 10 6 PBMC(experimental gruop 2). Survival rate within 4 weeks, white blood cell count, bone marrow nucleated cells (BMNC), granulocyte-macrophage colony forming unit(GM-CFU) and fibroblast colony forming unit (F-CFU) were examined. RESULTS: Survival rate, BMNC, GM-CFU, F-CFU were significantly higher in experimental group 2 than that in PBSCT group ( P

100 U/L were higher than those in group of CK-MB≤100 U/L(P

Objective: To assess the regional systolic and diastolic function of left ventricle by pulsed wave Doppler tissue imaging ( PW-DTI) in patients with coronary artery disease ( CAD ) . Methods : Seventy-seven cases of coronary angiography were adopted for study and grouped in terms of severity of coronary arterial lesion. Peak systolic, peak early diastolic, and peak late diastolic motion velocities (Sm, Em and Am, respectively) were recorded at 6 different sites on the basal and medial segments corresponding to the anteroseptal, posteroseptal, lateral, anterior, inferior, and posterior walls of the left ventricle with PW-DTI. Results: Am in segments corresponding to normal coronary arteries (group A) was lower than in segments corresponding to slightly stenosed coronaries (group B). There was no significant difference in Sm and Em between group A and group B. Compared with group A, Sm, Em and Am were significantly decreased in segments corresponding to severely stenosed coronary arteries ( group D). Conclusion : PW-DTI could be used to evaluate quantitatively and accurately the regional mvocardial function of left ventricle in patients with CAD.

Objective To discuss the correlation between aortic elasticity and coronary artery calcification by CT.Methods Totally 111 patients who were diagnosed of coronary artery disease underwent coronary artery CTA.The images were qualified for aortic elasticity measurement.All patients were divided into calcification negative group (n=43) and calcification positive group (n =68).The calcification positive group was further divided into light,medium,and serious groups according to their calcification scores.The ascending aortic images were reconstructed every 5 ％ R-R intervals.The cross-sectional areas and diameters of aortic in each R-R interval were measured automatically,then diameter variation rate (％ A0),aortic distensibility (A0D),aortic compliance (A0C) and aortic stiffness (A0SI) were calculated to evaluate aortic elasticity.Correlation between aortic elasticity and coronary artery calcification were analyzed.Results ％ A0,A0 D,A0C were lower and A0SI was higher in calcification positive group than those in calcification negative group (all P＜0.05).There was no significant differences in the four reference indexes of aortic elasticity among light,medium,and serious groups in calcification positive group (all P＞0.05).Correlation analysis demonstrated negative correlations between ％ A0,A0 D,A0 C and calcification scores,and a positive correlation between A0SI and calcification scores.Conclusion Aortic elasticity is correlated with coronary artery calcification,and the combination of them will be beneficial for the early diagnosis of coronary heart disease.

Objective To investigate the changes of sex hormones after lamivudine antiviral treatment in male hepa-titis B patients with compensated cirrhosis, and the significance of changes thereof. Methods Forty-six male patients with compensated cirrhosis were included in patient group. The patient group was given lamivudine antiviral treatment for 24 weeks. The sex hormone levels were compared between before and after treatment. Patients were divided into two groups (complete response group and incomplete response group) according to their response to the treatment. The sex hormone lev-els were compared between these two groups. Results (1)The level of PRL was significantly decreased, and the levels of T and E2 were significantly increased after lamivudine antiviral treatment in patient group.(2)There were significant differenc-es in levels of PRL, T and E2 between complete response group and incomplete response group. Conclusion (1)The lami-vudine antiviral treatment can improve the dysfunction of sex hormones.(2)There was a relationship between the changes of sex hormones and the responses of antiviral treatment.