Objective:To analyze the value of serum ceruloplasmin (CP) levels in predicting the outcome of patients with hepatitis B virus (HBV)-related acute-on-chronic liver failure (ACLF).Methods:The clinical data of 1 751 patients with HBV-ACLF treated in the Third Affiliated Hospital of Sun Yat-sen University from January 2010 to March 2018 were retrospectively analyzed. According to 30-day outcomes, 1 220 survival patients were classified into group A; 465 fatal patients and 46 patients receiving liver transplantation were classified into group B (total 531 cases). Risk factors associated with 30-day survival were estimated using Cox proportional hazards regression. ROC curve analysis was performed to evaluate the predictive value of CP on the 30-day outcome of patients with HBV-ACLF.Results:Multivariate analysis indicated that CP, albumin and alpha fetoprotein were independent protective factors for 30-day survival of HBV-ACLF patients ( P<0.05 or <0.01), while age, white blood cell count, AST, total bilirubin, INR, serum creatinine, HBV DNA, hepatorenal syndrome and hepatic encephalopathy were independent risk factors ( P<0.01). The area under the ROC curve (AUC) of CP was 0.570 (95% CI 0.540-0.599, P<0.01); while AUC of MELD score was 0.783 (95% CI 0.759-0.807, P<0.01) and MELD-Na score was 0.774 (95% CI 0.750-0.798, P<0.01). Compared with MELD score and MELD-Na score, the value of CP in predicting the 30-day prognosis of HBV-ACLF patients was lower ( P<0.01). The cut-off value of CP for predicting 30-day outcome of HBV-ACLF patients was 0.173 g/L, with the sensitivity of 69.4%, and the specificity of 41.6%. According to the cut-off value, the patients were divided into low CP level group (level of CP<0.173 g/L) and high CP level group (level of CP≥0.173 g/L); the 30-day cumulative survival rate of low CP level group was lower than that of high CP level group ( χ2=17.75, P<0.01). Conclusions:Serum CP level can predict the 30-day outcome of HBV-ACLF patients to a certain extent.

OBJECTIVETo construct an eukaryotic expression plasmid for AY358935 gene and explore its function.

METHODScDNA of the AY358935 gene was amplified by reverse transcription-PCR and cloned into pGEM-Teasy. The pGEM-T-AY was validated by sequencing and served as a template for the construction of eukaryotic expression plasmid. The pcDNA3.1-AY recombinant was validated by double enzyme digestion and used for transient transfection of M14 cells. Expression of the AY358935 protein and proliferation of the M14 cells were determined respectively by Western blotting and 3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-di-phenytetrazoliumromide (MTT) colorimetry.

RESULTSThe amplicons of RT-PCR were confirmed to have similar size with the cDNA fragment of the AY358935 gene as well as cloned region of pcDNA3.1-AY. The cloned region of pGEM-T-AY was sequenced to be identical with cDNA sequence of the AY358935 gene. M14 cells were transfected by the AY358935 gene, pcDNA3.1 and liposomes, respectively. After 48 h, expression of the AY358935 protein in M14 cells transfected with the AY358935 gene was significantly higher than other two groups. They also had a significantly higher absorbance value (A=0.74) than other two groups (A=0.39 and 0.46, respectively; P<0.05).

CONCLUSIONAn eukaryotic expression plasmid of the AY358935 gene was successfully constructed. Product of the AY358935 gene may promote the proliferation of M14 cells.

Objective To evaluate the efficacy and long-term safety of autologous bone marrow stem cells(ABMSC)transplantation in patients with hepatitis B virus(HBV)-associated decompensated liver cirrhosis.Methods This was an open-label,prospective matched case-control study.Thirty patients with HBV-associated decompensated liver cirrhosis hospitalized at the Third Affiliated Hospital of Sun Yat-Sen University from January 2005 to June 2010 were collected and infused with stem cells(stem cell group). Another thirty patients in control group were matched according to baseline characteristics and treated with standard medicine therapy.The patients in stem cell group were treated with stem cells infusion by hepatic artery or portal vein based on standard medicine therapy.All the patients were followed up for 5 to 10 years after surgery. Biochemical indicators were evaluated within the first 48 weeks after transplantation.The complications of cirrhosis and the cumulative incidence rate of hepatocellular carcinoma(HCC)were observed.Measurement data with normal distribution were analyzed by t test. Measurement data with non-normal distribution were compared by Mann-Whitney test.Count data were compared by χ2 test.The cumulative incidence rate of HCC development was compared by Kaplan-Meier analysis.Results The bone marrow aspiration and transplantation surgery were well tolerated in all patients in stem cell group.No complication related to stem cell transplantation therapy was observed. The levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),total bilirubin(TBil) and prothrombin time(PT)decreased,albumin level increased,while model for end-stage liver disease (MELD)scores decreased in both groups after treatment.Serum albumin level in stem cell group increased and ALT level decreased markedly at week 4,compared with that in control group at week 4(Z=2.188,P=0.029,Z=3.296,P=0.001,respectively).In stem cell group,21 patients received stem cells transplantation by hepatic artery and 9 patients by portal vein.Biochemical indicators were improved in all patients compared to baseline.However,there was no statistically significant differences between hepatic artery group and portal vein group.The median follow-up time was 6 years.Two patients in stem cell group and 1 patient in control group died(χ2=0.351,P=0.554).Six patients in stem cell group (20.0%)and 11 patients(36.7%)in control group developed HCC.There was no significant differences in the cumulative incidence rate of HCC between two groups(χ2= 0.148,P= 0.701).Hepatorenal syndrome did not development in either group.There were no statistically significant differences in the rates of complications including spontaneous peritonitis,hepatic encephalopathy and gastrointestinal hemorrhage between two groups after 5 to 10 years of follow-up(χ2=0.162,P=0.688,χ2=1.071,P=0.301,χ2=1.071,P=0.301,respectively).Conclusion ABMSC transplantation in patients with HBV-associated decompensated liver cirrhosis improves liver function transiently and has long-term safety.