Objective To explore the expressions of adiponectin and adiponectin receptor 2 (AdipoR2) in the liver of patients with hepatitis B virus (HBV)-related liver failure and their clinical implications in the pathogenesis of liver failure.Methods The healthy controls (HC),patients with chronic hepatitis B (CHB) and patients with HBV-related liver failure (HBV-LF) were enrolled in the study.Each group had 20 participants.The liver tissues from 11 CHB patients who were diagnosed by liver biopsy,6 patients with HBV-LF and 6 liver donors during liver transplantation were collected.Histological specimens were observed by hematoxylin-eosin staining and Masson trichrome staining under microscope.The mRNA and protein expressions of adiponectin and AdipoR2 in the liver were measured by semi-quantitative reverse transcription and polymerase chain reaction (SqRT-PCR) and immunohistochemical staining,respectively.The level of serum adiponectin was detected by enzymelinked immunsorbent assay.Serum biochemical parameters and HBV DNA levels were also measured.The pairwise comparison between groups was done by Student-Newman-Keuls and mann whitney U test.The relationship between two variables was analyzed using Spearman correlation.Results The level of serum adiponectin in HBV-LF group [(0.86 ± 0.15) ng/mL] was higher than those in HC [(0.59±0.15) ng/mL] and CHB groups [(0.62±0.13) ng/mL,Z=3.963,Z=3.774,both P＜0.01)],but showing no difference between CHB and HC groups (P＞0.05).The expressions of adiponectin and AdipoR2 mRNA in the liver were significantly higher in HBV-LF group (0.251 ±0.028 and 0.223 ± 0.021,respectively) than those in HC (0.091 ± 0.018 and 0.072 ± 0.020,respectively) and CHB (0.121±0.019 and 0.097±0.017,respectively) groups (q=18.428,17.069,19.807,18.673,respectively; all P＜0.01).Also,the expressions of adiponectin and AdipoR2 mRNA in CHB group were significantly higher than those in HC group (q=3.895,3.860,both P＜0.05).The expressions of adiponectin and AdipoR2 proteins in the liver were significantly higher in HBV-LF group (8.482±0.772 and 7.654±0.272,respectively) than those in HC (4.045± 0.815 and 2.804±0.623,respectively) and CHB (5.545±0.613 and 4.775±0.458,respectively) groups (q=15.327,11.542; Z=2.802,3.266; respectively; all P＜0.01).Also,the expressions of adiponectin,AdipoR2 proteins in CHB group were significantly higher than those in HC group (q=5.894,Z=3.266,both P＜0.01).In HBV-LF group,serum adiponectin levels as well as the expressions of adiponectin mRNA and protein in the liver were negatively correlated with serum albumin (r=-0.815,-0.886,-0.943; P＜0.01 or P＜0.05),but positively correlated with serum alanine aminotransferase (r=0.701,0.886,0.943; P＜0.01 or P＜0.05).The expression of AdipoR2 mRNA in the liver was negatively correlated with serum albumin (r=-0.943,P＜0.01),but positively correlated with serum aspartate aminotransferase (r=0.829,P＜0.05).Conclusions The expressions of adiponectin and AdipoR2 are up-regulated during HBV infection,especially in patients with HBV-LF,which might reflect the degree of necroinflammation in the liver.

ObjectiveTo investigate the effects and molecular mechanism of simvastatin in liver fibrosis model of non-alcoholic fatty liver disease (NAFLD) in vivo and hepatic stellate cell in vitro.MethodsFirstly,the rat liver fibrosis model of NAFLD was established by high-fat diet administration and intervened with simvastatin.The expression of endothelial nitric oxide synthase (eNOS),inducible nitric oxide synthase (iNOS) and Collagen Ⅰ at mRNA and protein level were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot.Secondly,quiescent phenotype of LX-2 cell line was induced by promoting adipocyte differentiation medium in vitro,and then the quiescent phenotype of LX-2 cell were treated with transforming growth factor β1(TGF-β1),Nitroso-L-arginine methyl ester (L-NAME) which was NOS inhibitor,simvastatin,TGFβ1 with simvastatin,and L-NAME with simvastatin separately.The changes of eNOS,iNOS,αsmooth muscle actin (α-SMA) and Collagen Ⅰexpressions at mRNA and protein level were determined by RT-PCR and Western blot.ResultsAs modeling time extended,the expressions of eNOS in rat's liver tissue of model group at mRNA and protein level decreased gradually,however the expression of iNOS and Collagen Ⅰ at mRNA and protein level increased gradually,compared with normal control group and the differences were statistically significant (P ＜0.05 and 0.01).By 24weeks,the expressions of eNOS in rat's liver tissue of simvastatin group at mRNA and protein level were increased,the expression of iNOS at mRNA and protein level were decreased and the expression of Collagen Ⅰ at mRNA and protein level were decreased (all P ＜0.05).The expression of eNOS in rat's liver tissue of model group negatively correlated with the expression of Collagen Ⅰ at mRNA and protein level (all P ＜0.01).The expression of iNOS positively correlated with that of Collagen Ⅰ at mRNA and protein level (all P ＜0.01).In LX-2 cell culture,L-NAME inhibited the activation of LX-2,reduced eNOS and iNOS expression and increased α-SMA and CollagenⅠexpression,consistent with the role of TGF-β1.Simvastatin could directly increase the eNOS expression both in quiescent and activated LX-2 cells,decrease iNOS expression,maintain quiescent phenotype and inhibit its activation.ConclusionsSimvastatin ameliorated the genesis and progression of liver fibrosis by increasing eNOS expression in LX-2 cells and reducing iNOS,α-SMA and Collagen Ⅰ expression.

Objective To investigate the effect of exogenous hydrogen sulfide on myocardial NF-κB signaling pathway in rats with hemorrhagic shock (HS).Methods Forty adult male rats,weighing 250-300 g,were randomly divided into 4 groups (n =10 each):sham operation group (Sham group),sham operation + NaHS group (Sham + NaHS group),HS group and HS + NaHS group.HS was induced by withdrawing blood from the femoral artery.After HS,NaHS 28 μmol/kg was injected intraperitoneally at 10 min before resuscitation in groups HS + NaHS and Sham + NaHS.MAP was monitored and recorded at 0,1.5,2,3,4 and 6 h after blood-letting.The rats were then sacrificed and hearts were removed for determination of phosphorylated IKKβ (pIKKβ),IκBα (pIκBα),NF-κB p65 (pNF-κB p65) and high-mobility group box 1 (HMGB1) and for examination of myocardial ultrastructure with light and electron microscope.Results Compared with Sham and Sham + NaSH groups,MAP was significantly decreased and the expression of pIKKβ,pIκBα,pNF-κB p65 and HMGB1 was up-regulated in HS and HS + NaHS groups (P ＜ 0.05).Compared with HS group,MAP was significantly increased and the expression of pIKKβ,pIκBα,pNF-κB p65 and HMGB1 was down-regulated in HS + NaHS group (P ＜ 0.05).The pathologic changes were attenuated in HS + NaHS group compared with group HS.Conclusion Exogenous hydrogen sulfide can attenuate myocardial injury induced by HS through inhibition of NF-κB signaling pathway and reduction of inflammatory response.

Objective To discuss the high-risk human papillomavirus (HPV )infection in the Guangxi .Methods A total of 26 796 copies of the test results for female cervical secretions specimens were analyzed retrospectively from what the second-genera-tion hybrid capture technology mixed detect 13 kinds of high-risk HPV viral DNA .Results Infection rate was 17 .94% in Guangxi , and the infection of HPV in north was higher than other regions ,and the differences of regions had statistically significant (P<0 .05) .The peak age of infection in the Guangxi region was less than 20 years old ,50- <60 years old ,and equal or more than 60 years old .The differences among age groups was statistically significant(P<0 .05) .Patients constitute HPV infection Guangxi re-gion had high viral load for 40 .93% ,and high viral load constituent ratio rise with age .Conclusion The existence of high-risk HPV infection is regional different and age different in the Guangxi region ,to high-prone areas ,infection high age should focus on monito-ring ,high viral load in patients with complete cervical cytology and histology examination if necessary .

OBJECTIVETo evaluate the efficacy of treatment with allogeneic peripheral blood stem cell transplantation for benzene-induced severe aplastic anemia.

METHODSHLA-compatible sibling (pregnancy) allogeneic peripheral blood stem cell transplantation (Allo-PBSCT) was successfully performed for a patient with severe aplastic anemia caused by benzene poisoning. 9.41 x 10(8) nucleated cells/kg, 12.49 x 10(6) CD(34) positive cells/kg and CFU-GM 8.2 x 10(5)/kg were infused. The patient was treated with cyclophosphamide (120 mg/kg), total body radiation (8 Gy) and anti-lymphocyte globulin (60 mg/kg) before transplantation. Donor buffy coat cells (9.02 x 10(8) nucleated cells/kg, 10.62 x 10(6) CD(34) positive cells/kg, 6.3 x 10(5) CFU-GM/kg) were infused again on day 18 after transplantation to prevent from graft failure. Graft versus host disease prophylaxis consisted of both methotrexate and cyclosporin A.

RESULTSThe lowest ANC was 0, the lowest platelet was 3 x 10(9)/L after transplantation. The patient achieved an ANC of greater than 0.5 x 10(9)/L from 21st day, and the platelet of greater than 50 x 10(9)/L from 28th day after transplantation. Grade I cGVHD was found the fourth month after grafting. Examination of recipient's bone marrow cells showed a normal 46, XX (presumably marrow donor) karyotype. Blood group changed from B to O.

CONCLUSIONThis is the first case reported in China showing a successful treatment of benzene-induced severe aplastic anemia with allo-PBSCT. Allo-PBSCT may be an effective remedy for this kind of patients.

Adult ; Anemia, Aplastic ; chemically induced ; therapy ; Benzene ; poisoning ; Humans ; Male ; Occupational Diseases ; chemically induced ; therapy ; Peripheral Blood Stem Cell Transplantation ; Transplantation, Homologous ; Treatment Outcome

The purpose of this study was to investigate the repair of the osteoarthritis(OA)-induced cartilage injury by transfecting the new TGF-β3 fusion protein (LAP-MMP-mTGF-β3) with targeted therapy function into the bone marrow-derived mesenchymal stem cells (MSCs) in rats. The recombinant of pIRES-EGFP-MMP was constructed by combination of DNA encoding MMP enzyme cutting site and eukaryotic expression vector pIRES-EGFP. LAP and mTGF-β3 fragments were obtained from rat embryos by RT-PCR and inserted into the upstream and downstream of MMP from pIRES-EGFP-MMP respectively, so as to construct the recombinant plasmid of pIRES-EGFP-LAP-MMP-mTGF-β3. pIRES-EGFP-LAP-MMP-mTGF-β3 was transfected into rat MSCs. The genetically modified MSCs were cultured in medium with MMP-1 or not. The transfected MSCs were transplanted in the rat OA models. The OA animal models were surgically induced by anterior cruciate ligament transaction (ACLT). The pathological changes were observed under a microscope by HE staining, Alcian blue, Safranin-fast Green and graded by Mankin's scale. pIRES-EGFP-LAP-MMP-mTGF-β3 was successfully constructed by means of enzyme cutting and sequencing, and the mTGF-β3 fusion protein (39 kD) was certified by Western blotting. Those genetically modified MSCs could differentiate into chondrocytes induced by MMP and secrete the relevant-matrix. The transfected MSCs could promote chondrogenesis and matrix production in rat OA models in vivo. It was concluded that a new fusion protein LAP-MMP-mTGF-β3 was constructed successfully by gene engineering, and could be used to repair the OA-induced cartilage injury.

AIMTo study the effect of recombinant hirudin (rH) on tPA-induced fibrinolysis and the possible mechanism of its action.

METHODSThe effect of rH on thrombin-fibrin complex (Th-Fn) was detected by 99mTc labeled rH. In the in vitro clot lysis, tPA as plasminogen activator, and recalcified plasma as plasminogen resource were used to study the influence of rH on fibrinolysis by detecting TAFIa, D-Dimer and FXIII.

RESULTSIn a canine model of femoral artery thrombosis, a clear radioactivity strip was imaged in 30 - 60 min on a part image, and the femoral vein thrombosis developed at 30 min. rH efficiently inhibited clot regeneration. Addition of TM could inhibit clot lysis obviously, and CPI could shorten the delay of clot lysis which due to TAFIa. There was a dose-dependent relationship with TM concentration and TAFI activation. FXIII activation was inhibited by low concentration of rH ( < or = 0.2 u x mL(-1)), and the level of fibrinolysis product, D-Dimer, increased.

CONCLUSIONrH could inhibit the thrombin binding to fibrin. rH inhibited the activation of TAFI and FXIII by combining with thrombin which resulted in enhancement of thrombolysis.

Animals ; Blood Coagulation ; drug effects ; Carboxypeptidase B2 ; metabolism ; Carboxypeptidases ; antagonists & inhibitors ; Dogs ; Factor XIII ; metabolism ; Femoral Artery ; Femoral Vein ; Fibrinolysis ; drug effects ; Fibrinolytic Agents ; pharmacology ; Hirudins ; genetics ; pharmacology ; Male ; Plant Proteins ; pharmacology ; Protease Inhibitors ; Recombinant Proteins ; pharmacology ; Thrombomodulin ; metabolism ; Thrombosis ; metabolism ; Venous Thrombosis ; metabolism

To explore a simple and effective method to determinate the volume of CD34(+) cells in the peripheral blood of donors received drug mobilization for stem cell transplantation by using flow cytometry, the mobilized peripheral blood from donors and 100 micro l fresh whole blood were labeled with monoclonal antibodies Anti-CD34-PE and Anti-CD45-FITC, after lying the red blood cells, and assessed with flow cytometer FL2 (log) vs SSC (log) and FL1 (log) vs SSC (log) were mainly used for analysis windows. The results showed that a level of CD34(+) cells in whole nucleated cells as low as 0.05% - 0.1% can be detected effectively using this method when 10(5) nucleated cells were counted. At day 5 or day 6, the level of CD34(+) cells in most samples of patients reached a peak volume, some of samples and the levels were more than one percent in. It was concluded that CD34(+) cells can be effectively determined by using this method. According to the relative rate of CD34(+) cells, the time to harvest the stem cells in blood can be determined.
Antigens, CD34 ; blood ; Blood Donors ; Flow Cytometry ; methods ; Hematopoietic Stem Cells ; cytology ; Humans