Analysis of NK Cells and Receptors in Peripheral Blood of Patients with HBV Related Acute-on-chronic Liver Failure in Different Phases / 中山大学学报(医学科学版)

[Objective]To investigate the character of expression of NK cells and its receptors in peripheral blood of pa-tients with HBV-related acute-on-chronic liver failure(HBV-ACLF)in different phase.[Methods]There are thirteen pa-tients with HBV-ACLF in advanced phase group and thirty-three patients in plateau phase group,with thirteen healthy per-son in control group.The frequency of NK cells and expression of NK cell activating receptors(NKAR)including NKG2D, NKp30 and NKp46,NK cell inhibitory receptors(NKIR))cludingsd ate the character of including NKG2A,KIR2DL1, KIR2DL3 and KIR3DL1,NK cell killing function related factors including Perforin,GranzymeB and FasL in peripheral blood of all patients were detected by flow cytometry.[Results]The percentage of NK cells in advanced group was lower than plateau group and healthy control group(H=7.771,P=0.021).The expression of KIR3DL1 in healthy control group was high-er than advanced group and plateau group(Z=6.639,P=0.036)while the expression of FasL was significantly lower than ad-vanced group and plateau group(Z=22.5,P<0.001).[Conclusion]Patients in advanced group had lower frequency of NK cells,lower expression of inhibitory receptor KIR3DL1 and higher expression of FasL than patients in plateau group and healthy control group,which is associated with immune status of patients in different phase of liver failure.

Relationship between the genetic variation in interleukin 28B and response to antiviral therapy in patients with chronic hepatitis C / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Genetic variations at the interleukin 28B (IL-28B) locus are important in predicting outcome following therapy for chronic hepatitis C virus (HCV) infection. The aim of this research was to evaluate the role of IL-28B single nucleotide polymorphism (SNP) variations in Chinese patients undergoing pegylated interferon-α plus ribavirin (PEG-IFN-α/RBV) treatment.</p><p><b>METHODS</b>To determine the effect of IL-28B variation on the response to HCV therapy, these variants were genotyped in a cohort of 220 patients who were chronically infected with HCV and received combined PEG-IFN-α/RBV therapy.</p><p><b>RESULTS</b>The proportions of rs12979860 CC, CT, and TT genotypes were 71.4%, 25.0%, and 3.6% respectively, in the sustained virological response (SVR) group; 15.8%, 60.5%, and 23.7% respectively, in the null virological response (NVR) group; and 38.1%, 52.4%, and 9.5% respectively, in the relapse (Rel) group (P < 0.05). Logistic regression analysis showed that, compared to those having the CC genotype, CT heterozygotes had an increased risk of NVR and Rel (OR = 10.95, 95%CI = 4.12-29.11, P = 1.5×10(-7) and OR = 3.93, 95%CI = 1.86-8.32, P = 2.1×10(-4) respectively). The RNA quantification assay showed that patients with genotype CC exhibited much higher levels of IL-28 expression than those with genotype CT or TT (P < 0.001).</p><p><b>CONCLUSIONS</b>The IL-28B SNP rs12979860 genotype was related to the effectiveness of HCV therapy: patients with the CC rs12979860 genotype had higher rates of SVR than those with the CT or TT genotype, and the CC genotype revealed a significantly higher level of IL-28 mRNA expression.</p>

Safety evaluation and quality control of MSCs from hepatitis B patient in vitro / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To get mesenchymal stem cells (MSCs) from hepatitis B patient and to valuate the safety and quality after long-term culture in vitro.</p><p><b>METHODS</b>The cells obtained directly from bone marrow and cultured in Mesen Pro medium supplemented with FGF, and the morphology of MSCs was observed. Surface antigens of the MSCs were analyzed by flow-cytometry. The bacteria, virus, endotoxin and residual serum of cell suspension were detected. The MSCs and perpheral blood T lymphocytes were co-cultured in 48 well plates for 72 h and the T lymphocyte proliferation was measured by using MTT reduction method and the effect of MSCs on T lymphocyte transformation stimulated by PHA was also observed. The oncogenicity of MSCs was verified by the tumorigenesis test in sofo agar. The genetic stability of MSCs was examined by karyotype analysis.</p><p><b>RESULT</b>The MSCs from hepatitis B patient could be passaged to many generations and had strong abilities of proliferation. They expressed stem cell-surface antigens and maintained normal karyotype, prevented the pollution of bacteria and viruses, inhibited the immune response of allogenic T lymphocytes and no oncogenicity found.</p><p><b>CONCLUSION</b>The MSCs have proliferative potentials, can be passaged in long-term cultures in Mesen Pro medium without oncogenicity, can maintain normal karyotype, can inhibit the immune response of T lymphocytes and can alleviate the grafe-versus diseases. The MSCs can be served as a new type of cells in cell and gene therapy.</p>

Establishment and application of standard serum proteomic pattern for diagnosis of early liver cancer / 南方医科大学学报

<p><b>OBJECTIVE</b>To establish standard serum proteomic pattern for early liver cancer and explore its value in early liver cancer diagnosis.</p><p><b>METHODS</b>The serum samples were obtained from 12 healthy subjects and 12 patients with early liver cancer. After removal of serum albumin and IgG, 300 µg serum sample was mixed with the hydrating solution for two-dimensional gel electrophoresis. Each experiment was repeated for 3 times. With silver staining and image analysis, the differentially expressed proteins were identified to establish the standard proteomic pattern for early liver cancer. The proteomics pattern was then tested in the diagnosis of 13 cases of chronic liver disease and 10 healthy individuals in a double-blind manner.</p><p><b>RESULTS</b>Thirty-eight differentially expressed protein spots were identified. Eleven protein spots with differential expressions by over 5 folds were analyzed to establish the standard proteomics pattern for early liver cancer, which showed a sensitivity of 92% and specificity of 90% in early diagnosis of liver cancer.</p><p><b>CONCLUSION</b>The standard serum 2-DE proteomic pattern for early liver cancer shows distinct differences from that of healthy subjects. The differentially expressed proteins and their specific combination pattern may have the potential for diagnosis and prognostic evaluation of early liver cancer.</p>

Study on HBV antigens and IL-12 affecting T cell-mediated immunity in HBsAg carriers / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the effect of HBV antigens and pathological mechanism of chronic HBV infection by analyzing the cellular immune function of peripheral blood mononuclear cells (PBMCs) from HBsAg carriers.</p><p><b>METHODS</b>PBMCs were prepared from individuals with chronic asymptomatic HBV infection and cultured in the presence of different antigens and/ or cytokines. The levels of cytokines in culture supernatants were detected by ELISA method. The phenotype of the cells was detected by FACS.</p><p><b>RESULTS</b>The levels of IFN y secreted by PBMCs from HBsAg carriers were (48.3+/-19.8) pg/ml, significantly lower than that from healthy controls (t = 3.023, P less than 0.05); The IFN y produced by PBMCs from HBeAg positive patients due to HBsAg and HBcAg stimulation were (50.4+/-51.6) pg/ml and (63.2+/-36.9) pg/ml, significantly lower than that of HBeAg negative patients (t = 2.468 and 3.184, P less than 0.05, respectively). The IL-12p70 secreted by PBMCs from HBeAg positive patients was also significantly lower than that of HBeAg negative patients (P less than 0.05); Exogenous IL-12 promoted significantly PBMCs to secrete IFN y (P less than 0.01) and IL-12 combined with HBV antigens activated CD8+CD45RA+CCR7+ and CD8+CD45RA-CD62L+ cells. IL-12 secreted by PBMCs decreased in HBeAg positive patients, which may be the crucial reason of viral persistence in chronic HBV carriers. Exogenous IL-12 combined with specific HBV antigen could promote the central memory CD8+ T cells to produce IFN y.</p>

Programmed death-1 (PD-1) and PD-L1 expression during antiviral treatment of chronic hepatitis B / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To study PD-1 and PD-L1 expressions during 24 weeks telbivudine antiviral treatment in patients with chronic hepatitis B (CHB) and to explore the relationship between PD-1 expression and HBeAg/HBeAb seroconversion.</p><p><b>METHODS</b>Ten CHB cases with HLA-A2 and HBeAg positive were treated with telbivudine 600 mg/d orally for 24 weeks. Fresh blood samples were collected at week 0, 12 and 24 after treatment. HBV-specific CD8+ T cells were expanded in vitro. Cell culture medium were collected for interferon gamma (IFNgamma) detection. Flow cytometry was used to detect the HLA-A type, PD-1, PD-L1 and HBV specific CD8+ T cells. The expressions of PD-1 and PD-L1, the counts of HBV-specific CD8+ T cells in circulating CD8+ lymphocytes, and IFNgamma concentration in culture medium were evaluated during antiviral treatment.</p><p><b>RESULTS</b>At week 0, 12 and 24 after telbivudine treatment, 7 of 10 patients were HBV DNA undetectable, 2 were HBeAg seroconversion and 2 were HBeAg lose but anti-HBe negative. The frequency of PD-1-positive PBMCs were 52.1%+/-17.0%, 39.1%+/-18.2% and 23.4%+/-16.3% (week 24 vs week 0, P < 0.01) respectively; PD-L1 positive PBMCs were 45.6%+/-15.4%, 34.6%+/-16.2% and 20.9%+/-9.5% respectively(week 24 vs week 0, P < 0.01; week 24 vs week 12, P < 0.05). The frequency of PD-1-positive CD8+ T cells were 76.2%+/-10.4%, 66.5%+/-15.4% and 49.5%+/-25.3% respectively (week 24 vs week 0, P < 0.01; week 12 vs week 0, P < 0.05; week 24 vs week 12, P < 0.05); HBV-specific CD8 cells were 1.3%+/-0.5%, 1.5%+/-1.0% and 2.2%+/-1.5%; IFNgamma levels in cell culture medium were (91.7+/-82.1) pg/ml, (99.4+/-93.5) pg/ml and (109.5+/-86.6) pg/ml. A remarkable decrease of PD-1 and PD-L1 expressions and increase of HBV-specific CD8+ T cells were observed in patients who had HBeAg/HBeAb seroconversion at week 24.</p><p><b>CONCLUSIONS</b>Direct suppression of HBV replication by telbivudine in CHB patients can decrease PD-1 and PD-L1 expressions and restore HBV-specific CD8+T cells. The relationship between the changes of PD-1 expression and HBeAg/HBeAb seroconversion during antiviral therapy in HBeAg-positive patients need to confirm by future study.</p>

The effects of diethylene glycol on liver function / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the hepatotoxic effects of accidental intravenous diethylene glycol (DEG.) poisoning in patients with liver disease.</p><p><b>METHODS</b>Clinical data and liver function results were obtained from 64 patients with liver diseases who had been accidentally treated with diethyl glycol-contaminated agent and 45 cases with hepatorenal failure. The hepatotoxic effects of diethylene glycol DEG on the patients with liver diseases were assessed by multivariable logistical regression analysis.</p><p><b>RESULTS</b>Of the 64 cases with liver diseases, 15 cases (23.4%) developed toxic presentations following the accidental administration of DEG. All affected cases were male. Twelve of the 15 poisoned patients (80%), died within 7 days of exposure to DEG. The most common clinical manifestations included kidney damage, renal failure, metabolic acidosis, and nerve system disturbances. The intravenous administration of DEG resulted in only mild liver function impairment. In terms of risk factors, both gender (r = 4.266, P less than 0.05) and the severity of jaundice prior to DEG administration were related to the occurrence of toxin-induced renal failure (r = 7.640, P less than 0.01).</p><p><b>CONCLUSIONS</b>DEG may worsen liver damage in patients with liver diseases.</p>

Detection of Helicobacter species related genes coding for 16S rRNA in the liver tissue of patients with chronic liver disease / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the infection of Helicobacter pylori (Hp) in the liver tissue of patients with chronic liver disease and the association between Hp and chronic liver disease.</p><p><b>METHODS</b>Liver tissue samples were obtained by liver biopsy and surgical resection from 30 healthy subjects, 30 patients with chronic hepatitis, 30 with cirrhosis and 30 with liver cancer. All the samples were confirmed by pathological examination. The gene fragment coding for 16SrRNA were amplified by PCR with sequence analysis.</p><p><b>RESULTS</b>The PCR product of the 16SrRNA gene was 109 bp in length. Hp 16SrRNA was detected in 18 out of 30 liver biopsy samples from patients with primary cancer (60.0%), in 14 samples from patients with liver cirrhosis (47.0%), and in none of the samples from normal subjects or patients with chronic hepatitis. Sequencing analysis of Hp 16SrRNA gene in the liver tissue showed a 98.8% homology with the gene fragment encoding Hp 16SrRNA.</p><p><b>CONCLUSION</b>Hp is identified in the liver tissue of patients with chronic liver disease, suggesting the possible correlation between Hp infection and hepatocellular carcinoma.</p>

Status of H. pylori infection in the hepatic tissue of patients with hepatocarcinoma / 南方医科大学学报

<p><b>OBJECTIVE</b>To survey the status of Helicobacter pylori (Hp) infection in the liver of patients with liver cancer and explore the correlation between Hp infection and liver cancer.</p><p><b>METHODS</b>Liver specimens were obtained 30 patients with chronic liver disease (CLD), 30 with liver cancer, and 30 healthy control subjects. The specimens were cultured in Colombian agar medium in microaerophilic condition for isolation of Hp. The presence of Hp was also identified by biochemical test, urease test, immunohistochemistry, and PCR and sequence analysis.</p><p><b>RESULTS</b>Hp was found in none of the liver specimens by culture tests. One out of the 30 patients with liver cancer (3%) was positive for Hp by urease tests. Immunohistochemical staining identified Hp positivity in 9 liver cancer tissue (30.0%), but in none of control or CLD cases (P<0.05). PCR detected Hp 16S rRNA in 18 out of the 30 patients with liver cirrhosis (60.0%) and in none of the control or chronic hepatitis cases. Sequencing analysis of Hp 16S rRNA in liver tissue showed a 98.8% homology with the Hp16S rRNA gene.</p><p><b>CONCLUSION</b>A greater incidence of Hp infection in the liver is identified in patients with liver cancer than in healthy subjects and CLD patients, suggesting a potential correlation between Hp infection to liver cancer.</p>

Clinical features, laboratory findings and imaging appearances of venous diethylene glycol poisoning in patients with liver disease / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>There was a hospital outbreak of venous diethylene glycol poisoning in Guangzhou, China. It is the only massive episode of venous diethylene glycol poisoning in history. Here we report its clinical features, laboratory findings, and imaging appearances.</p><p><b>METHODS</b>The clinical features of 15 venous diethylene glycol poisoning patients with liver disease were analyzed and summarized. Their laboratory findings and imaging appearances were comparatively analyzed before and after poisoning.</p><p><b>RESULTS</b>All poisoned patients presented with oliguric acute renal failure with anuria after a mean of 6 days. Carbon dioxide combination power of 13 patients dropped after a mean of 9 days with valley value on the 10th day, when metabolic acidosis developed. Gastroenteric symptoms or aggravation of gastroenteric symptoms were displayed in 11 patients after a mean of 9 days. Neurological system impairment was observed in 10 patients after a mean of 14 days. Seven patients had low fever after a mean of 6 days. Causes of death of 14 patients included multiple organ dysfunction syndrome, severe lung infection and massive haemorrhage of digestive tract. Blood creatinine and urea nitrogen were abnormal after a mean of 5 days with peak value on the 11th and 14th days, respectively. Serum calcium had no obvious change, and phosphorus was distinctively increased. Liver functions did not change significantly. Poisoned patients had higher white blood cell counts, but lower red blood cell counts and hemoglobin value. Of the 7 patients who exhibited mild, moderate or severe patchy consolidation shadowing in the lung, 2 manifested mild or severe gaseous distention and dilation of gastroenteric tract.</p><p><b>CONCLUSIONS</b>Main features of venous diethylene glycol poisoning in patients with liver disease include oliguric acute renal failure, metabolic acidosis, gastroenteric symptoms or aggravation of gastroenteric symptoms, neurological system impairment and low fever, with a mortality rate of 93.33% in poisoned patients. There is also higher white blood cell counts and anemia, patchy consolidation shadowing in the lung, gaseous distention and dilation of gastroenteric tract, which occurs later than mild patchy consolidation shadowing and earlier than moderate patchy consolidation shadowing in the lung.</p>

The predictive value of ALT, HBeAg and HBV DNA levels at baseline and the degree of HBV suppression at week 12 adefovir dipivoxil treatment to the efficacy of it at week 52 in patients with HBeAg-positive chronic hepatitis B / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To study the predictive value of ALT, HBeAg and HBV DNA levels at baseline and HBV DNA levels at week 12 adefovir dipivoxil (ADV) treatment to the efficacy of it at week 52 in patients with HBeAg-positive chronic hepatitis B (CHB).</p><p><b>METHODS</b>Ninety-eight HBeAg-positive CHB patients with serum HBV DNA>or=1x10(6) copies/ml and ALT levels between 1.5 to 10 times of upper limits of normal (ULN) were enrolled in the study. Ten mg/d of ADV was administered for 52 weeks. Line serum samples were collected for measuring HBV DNA and HBV markers. The efficacy of the treatment at week 52 was evaluated in patients with different ALT, HBeAg and HBV DNA levels at baseline and HBV DNA levels at week 12 after treatment.</p><p><b>RESULTS</b>At week 52 of ADV treatment, the rates of HBV DNA<10(3) were 72.7%, 66.7% and 53.0% respectively in patients with ALT>5xULN, HBeAg<or=350 s/co and HBV DNA<or=10(8) copies/ml. Significant differences were found between them and patients with ALT<2xULN (38.0%, P<0.05), HBeAg>350 s/co (30.2%, P<0.01) and HBV DNA>10(8) copies/ml (34.4%, P<0.05) at baseline. HBeAg seroconversion rates were 42.2% and 7.5% (P<0.01) in patients with HBeAg titer<or=350 and >350 S/co at baseline. In patients with HBV DNA<10(3), 10(3)-10(5) and >10(5) copies/ml at week 12, the ratios of them with HBV DNA<10(3) less than 1000 copies/ml at week 52 were 82.6%, 57.1% and 17.5% and significant differences were found between these groups (P<0.05); HBeAg seroconversion rates were 52.2%, 25.7% and 5.0% (P<0.05); ALT normalization rates were 100%, 83% and 75%, significantly higher in patients with HBV DNA<10(3) copies/ml than those with HBV DNA>10(5) copies/ml (P<0.05) at week 12. HBV DNA and HBeAg seroconversion at week 52 correlated with HBV DNA levels at week 12 (r=0.6 and r=0.5 respectively, P<0.01).</p><p><b>CONCLUSIONS</b>In HBeAg-positive CHB patients treated with adefovir dipivoxil, HBV DNA levels at week 12 can be used to predict the efficacy at week 52. HBV DNA<10(3) copies/ml at week 12 predict a better treatment result at week 52.</p>

A multicenter clinical study of nafate for treatment of hepatic fibrosis in patients with chronic liver diseases / 中华肝脏病杂志

<p><b>OBJECTIVES</b>To investigate the effect of nafate (compound fetal cow liver extract tablets) in hepatic fibrosis patients with chronic liver diseases.</p><p><b>METHODS</b>One hundred fifteen hepatic fibrosis patients with chronic liver diseases from 3 medical centers were included in this study. All patients were given nafate orally for twenty-four weeks. Before treatment and 12, 24 and 36 weeks after the treatment, serum levels of hyaluronic acid (HA), laminin (LN) and type IV collagen (IV-C) of the patients were measured by radioimmunoassay and their liver biopsy specimens were also assessed.</p><p><b>RESULTS</b>Before treatment and 24 and 36 weeks after the nafate treatment, serum levels of HA were (279.2+/-81.4) ng/ml, (136.8+/-56.7) ng/ml and (86.9+/-40.7) ng/ml respectively, serum levels of LN were (170.8+/-73.0) ng/ml, (112.5+/-39.5) ng/ml, and (60.8+/-31.8) ng/ml respectively, and IV-C levels were (153.7+/-60.1) ng/ml, (112.4+/-43.1) ng/ml, and (96.3+/-44.1) ng/ml respectively. There was a significant reduction in these serum values after the treatments. Histopathological examinations of the liver biopsies showed that the degree of hepatic fibrosis obviously declined by one or two degrees after the treatments.</p><p><b>CONCLUSIONS</b>nafate has positive effects in treating hepatic fibrosis.</p>

Venous diethylene glycol poisoning in patients with preexisting severe liver disease:a clinical analysis / 中华传染病杂志

Objective To describe the clinical features of venous diethylene glycol poisoning and to identify factors correlating with such kind of poisoning.Methods Retrospective chart review was performed to analyze the epidemiology,clinical presentation,hepatorenal functions,bemodynam- ics and pathological characteristics of 64 patients with severe liver diseases who received intravenous diethylene glycol.Comparative analyses of correlating factors and causes of poisoning were based on the presence or absence of poisoning.Results Fifteen cases of poisoning were reported.After a 5 day incubation period,all poisoned patients displayed acute renal failure and 11 cases with digestive tract symptoms and(or) symptom exacerbations were noted.Neurological system impairment was observed in 10 cases after 2 weeks.Metabolic acidosis developed in 13 cases.Poisoned patients exhibited signif- icantly lower red blood celI(RBC)[(2.32?0.76)?10~(12)/L],hemoglobin(Hb) [(79.5?23.6)g/L] value and higher white blood cell(WBC)[(9.78?3.75)?10~9/L] count.Renal biopsy of poisoned patients revealed acute tubular necrosis and interstitial nephritis.Twelve poisoned patients died.Sig nifieant differences were found between groups regarding preexisting severe hepatitis,ascites,renal disease and diuretic therapy.Prior to diethylene glycol injections,mean values of neutrophil,blood urea nitrogen(BUN),creatinine(Cr) and calcium and phosphorousions differed significantly between groups.Conclusions Features of venous diethylene glycol poisoning include oliguric acute renal fail- ure,metabolic acidosis,digestive symptoms,nervous system impairment and a high probability of anemia and WBC proliferation.Mortality is high.Correlative factors include preexisting severe liver disease,renal disease and infection.