The oral glucose tolerance test,insulin release test and C peptide relea se test were carried out in 69 hypertensive patients and 55 normotensive subjects. It was found that,the concentrations of blood glucose,serum insulin and C peptide and the occurrence rates of glucose metabolic abnormality(diabetes mellitus and impaired glucose tolerance)of hypertensive group were higher than those of normotensive control group.The co-existence of hyperglycemia and hyperinsulinemia suggests the existence of insulin resistance.

OBJECTIVETo study the mechanism of hepatitis B virus infected patients who is negative for HbsAg.

METHODSDNA sequences of 46 patients were analyzed. In these patients, HBsAg was negative but HBV DNA was positive and six new HBsAg variants were identified. Four of the six variants were combined point mutants and two were insertion variants. These S genes were subcloned into eukaryotic expression vector EBO-plpp, and the recombinant eukaryotic expression plasmids were transfected into COS7 cells. Cell lines expressing mutant type HBsAg were obtained. The supernatants were detected by ELISA and RIA.

RESULTSOnly the two-amino acid-insertion variants could be detected and the others failed to react with polyclonal and monoclonal antibodies against HbsAg.

CONCLUSIONThe results indicated that the point mutations and insertions may result in a conformational change of the S gene, which affect HBsAg antigenicity, suggesting a possible relationship between the variants and the negative conversion of HBsAg of the patients.

Animals ; Antigenic Variation ; COS Cells ; Cercopithecus aethiops ; Hepatitis B Surface Antigens ; genetics ; immunology ; Hepatitis B virus ; genetics ; immunology ; Hepatitis B, Chronic ; immunology ; virology ; Humans ; Plasmids ; genetics ; Point Mutation ; Transfection

BACKGROUNDIt is still unclear whether viral genetic variability influences response to interferon (IFN)-alpha treatment. Recent reports suggest that IFN-alpha effects may be associated with hepatitis B virus (HBV) post-transcriptional regulation. This study was designed to explore the heterogeneity of HBV post-transcriptional regulatory elements (HPRE) and the relationship between the diversity of HPRE and the response to IFN-alpha treatment.

METHODSThe HPRE sequences from 31 Chinese patients infected with HBV were determined by directly sequencing of polymerase chain reaction (PCR) product, and comparing them to those from Caucasian patients. Subsequently, eukaryotic expression vectors containing HPRE at various points were constructed and transfected into HepG2 cells, which were then exposed to recombinant human cytokines.

RESULTSThe T to C point mutation at nt 1504 and the C to T (G) at nt 1508 in HPRE were found in 21 and 19 patients with chronic hepatitis B, respectively; the C to T point mutation at nt 1509 was found in 17 patients. These point mutations did not exist in the HPRE of the Caucasian patients. The activity of the CAT gene obviously increased in the case of T to C point mutation at nt 1504, but did not change in the case of the C to T (G) mutations at nt 1508 and 1509. The activity of the CAT gene at these point mutations of HPRE could be inhibited by IFN-alpha/gamma and tumor necrosis factor (TNF)-alpha except for the point mutations at nt 1508 of HPRE which may escape the suppression role of IFN-alpha on HPRE.

CONCLUSIONSThere are point mutations between the HPRE of Chinese and Caucasian HBV patients, which might be correlated with response to IFN-alpha. The variation of HPRE might affect the function of HPRE and influence the regulative function of IFN-alpha other than that of IFN-gamma or TNF-alpha on HPRE.

Chloramphenicol O-Acetyltransferase ; metabolism ; Genes, Regulator ; Hepatitis B virus ; drug effects ; genetics ; Hepatitis B, Chronic ; drug therapy ; virology ; Humans ; Interferon-alpha ; pharmacology ; Interferon-gamma ; pharmacology ; Plasmids ; Point Mutation ; Tumor Necrosis Factor-alpha ; pharmacology

BACKGROUNDSimultaneous pancreas-kidney transplantation (SPKT) frees the diabetic patient with end-stage nephropathy from dialysis and daily insulin injections. Herein, we review consecutive cases of SPKT with bladder drainage performed at our institution over an 8-year period.

METHODSThe study population included 21 patients (16 males and 5 females) who underwent SPKT between September 2001 and September 2009. Seven patients had type-1 diabetes and 14 had type-2 diabetes. Nineteen patients were on dialysis at the time of transplantation. Donation after cardiac death donors were selected for SPKT. The mean human leukocyte antigen match was 2 (range 0 - 4). SPKT was always performed using bladder drainage and vascular anastomoses to the systemic circulation. Immunosuppressive treatment consisted of anti-lymphocyte globulin induction followed by tacrolimus, mycophenolate mofetil, and prednisone.

RESULTSThe mean hospital stay was 45.43 days. After a mean follow-up of 39.4 months, survival rates for patient, kidney, and pancreas were 76.2%, 76.2%, and 66.7% at 1 year; 76.2%, 59.3%, and 55.6% at 5 years; and 57.1%, 39.5%, and 41.7% at 8 years, respectively. Major complications included anastomotic leaks, reflux pancreatitis, and rejection. Six patients died from septic shock (n = 3), duodenal stump leak (1), cardiac arrest (1), or renal failure (1). Eight kidney grafts were lost due to acute rejection (n = 2), chronic rejection (3), and death with a functioning graft (3). Pancreatic graft failure (9) was caused by thrombosis (n = 1), rejection (2), duodenal stump leak (1), and death with a functioning graft (5).

CONCLUSIONSSPKT is a valid therapeutic option for uremic diabetics although few hospitals in China can undertake SPKT.

Adult ; Diabetes Mellitus, Type 1 ; surgery ; Diabetes Mellitus, Type 2 ; surgery ; Female ; Graft Rejection ; Humans ; Immunosuppressive Agents ; therapeutic use ; Kidney Transplantation ; adverse effects ; mortality ; statistics & numerical data ; Male ; Middle Aged ; Pancreas Transplantation ; adverse effects ; mortality ; statistics & numerical data ; Postoperative Complications ; Retrospective Studies ; Treatment Outcome ; Urinary Catheterization

OBJECTIVETo investigate the clinical significance of neutralizing anti-interferon-alpha antibodies (NA) in chronic hepatitis B patients treated with recombinant interferon-alpha(rIFN-alpha).

METHODSOne hundred and eighty-one patients (128 male and 53 female) with histological proven chronic hepatitis B were treated with 5 MU recombinant interferon-alpha 1b (rIFN-alpha 1b) subcutaneously thrice weekly for 6 to 37 (median 10) months. For each patient, Specific detection of serum HBV DNA level with fluorescent-quantitative PCR, HBeAg with enzymoimmunoassay and NA with an antiviral neutralizing biological assay were performed during therapy.

RESULTSNA was found in 61 (33.7%) of 181 patients. At the end of treatment, complete-response was achieved in 17 (27.9%) of 61 patients with NA and in 54 (45.0%) of 120 patients without NA, respectively (chi2=4.979). For NA positive patients, the complete-response rate was significantly lower in those who had not achieved partial-response prior to or at the same time as NA occurred than in those who did (3.8%, 1/26, vs. 45.7%, 16/35, chi2 = 7.457). Moreover, it was lower in patients who either had 20pg/ml of serum HBV DNA or above or HBV DNA had being reduced by less than 60% recent 3 months, but higher in those who had less than 20pg/ml of HBV DNA and HBV DNA had being reduced by 60% or above (20.0%, 9/45, vs. 56.3%, 9/16, chi2 = 11.009).

CONCLUSIONNA may negate the antiviral effects of rIFN-alpha in chronic hepatitis B patients treated with rIFN-alpha, especially if they appear before partial-response or at the occasion at which serum HBV DNA level was not below 20pg/ml or HBV DNA had being reduced by less than 60% recent 3 months.

Antibodies ; blood ; DNA, Viral ; blood ; Female ; Hepatitis B, Chronic ; drug therapy ; virology ; Humans ; Interferon-alpha ; immunology ; therapeutic use ; Male ; Recombinant Proteins ; therapeutic use

Hepatitis B Core Antigens ; immunology ; Hepatitis B virus ; genetics ; immunology ; Hepatitis B, Chronic ; virology ; Humans ; T-Lymphocytes, Cytotoxic ; immunology

OBJECTIVETo investigate the relationship of virological breakthrough and production of neutralizing anti-interferon antibody (NAb) in chronic hepatitis B patients treated with recombinant interferon-alpha (rIFN-alpha).

METHODFour hundred eighty-five patients with histological proven chronic hepatitis B were treated with 5 MU recombinant interferon-alpha 1b (rIFN-alpha1b) thrice weekly for 6-37 months (median 10). Serum HBV DNA, HBeAg and NAb levels of the patients were detected by fluorescent-quantitative PCR, enzymoimmunoassay and antiviral neutralizing biological assay respectively during the therapy.

RESULTSVirological breakthrough occurred in 66 patients (13.6%), and NAb was found in 98 patients (20.2%) of the total 485 patients. The rate of NAb positivity was higher in patients with viral breakthrough than those without it (68.2%, 45/66, vs 12.6%, 53/419, chi(2)=109.06, P < 0.01), and viral breakthrough occurred more in patients with positive NAb than with negative NAb (45.9%, 45/98, vs 5.4%, 21/387, chi(2)=109.06, P < 0.01). The time of the viral breakthrough occurrence and the time of NAb production had a significant correlation (P < 0.01). The occurrence of viral breakthrough was also influenced by the age of patients (P < 0.05) and HBeAg status (P < 0.01) before they were treated.

CONCLUSIONViral breakthrough occurred in 13.6% of our 485 chronic hepatitis B patients treated with recombinant interferon-alpha. Their viral breakthrough and production of NAb production had a significant correlation.

Adult ; Antibodies, Neutralizing ; biosynthesis ; Female ; Hepatitis B Antibodies ; biosynthesis ; Hepatitis B virus ; immunology ; Hepatitis B, Chronic ; drug therapy ; virology ; Humans ; Interferon Type I ; therapeutic use ; Male ; Recombinant Proteins ; Young Adult

OBJECTIVETo study the cloning and expression of flagellin gene from Chinese Borrelia burgdorferi, PD91 strain and to evaluate the feasibility of using recombinant protein as diagnostic antigen when comparing the gene sequence with flagellin gene from North American Borrelia burgdorferi B31.

METHODSThe piece of genes coding flagellin from Chinese Borrelia burgdorferi PD91 by polymerase chain reaction (PCR) method was obtained, and constructed recombinant plasmid, before transformed into E. coli BL21 strain, and induced. The recombinant plasmid was identified with enzyme cutoff and gene sequence comparison. Efficient expression strain was selected and the expression product was analyzed with sodium amplified polymorphic-polyacrylamide gel electrophoresis (SDS-PAGE) and Western-blot method.

RESULTSThe recombinant protein (r-flagellin) expressed in host bacteria was successful. By means of western-blot assay, the immunological response showed the same antigenicity between r-flagellin and PD91 flagellin. The piece of genes coding flagellin of PD91 was 1011 bp, but when comparing with that of North American Borrelia burgdorferi it showed 94.70% homology. Homology between the sequence of amino acid of the r-flagellin and that of B31 flagellin was 95.85%.

CONCLUSIONFlagellin gene of Borrelia garinii of Chinese Lyme disease spirochete was successfully cloned and expressed for the first time. It was proved that the immunoreactivity of r-flagellin was the same as the natural flagellin.

Amino Acid Sequence ; Base Sequence ; Borrelia burgdorferi ; genetics ; isolation & purification ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Flagellin ; biosynthesis ; genetics ; Humans ; Lyme Disease ; microbiology ; Molecular Sequence Data ; Plasmids ; genetics ; Recombinant Proteins ; biosynthesis ; genetics

Epitopes, T-Lymphocyte ; HLA-A2 Antigen ; immunology ; Hepatitis B Core Antigens ; genetics ; Hepatitis B virus ; genetics ; immunology ; Humans ; Mutation

OBJECTIVETo investigate the prevalence of three lamivudine-resistant HBV mutants in lamivudine-treated Chinese patients.

METHODSUsing three pairs of of HBV polymerase gene B and C domain fragments were amplified by PCR. The PCR products were then digested by restriction enzyme Nde I and Nla III. The digested products were analyzed by electrophoresis. With this method, the prevalence of the three lamivudine-resistant mutants in lamivudine-treated Chinese patients was investigated.

RESULTSAfter Nde I digestion of p24 and p29 amplified product, HBV wild type could be easily separated from YMDD mutant. At the same time, YIDD could be separated from YVDD mutant after Nla III digestion of p24 and p29 amplified product. By this method, the authors found that these eleven patients were infected with lamivudine-resistant mutants. Six of them were infected with M5501 mutant; five were infected with M550V mutant (one of them had both M550V and L526M mutations).

CONCLUSIONThe method of the present study was demonstrated to be an easy way to detect HBV lamivudine-resistant mutants and can be applied to clinical monitoring of lamivudine resistance.

Adolescent ; Adult ; Antiviral Agents ; pharmacology ; therapeutic use ; DNA Mutational Analysis ; DNA Primers ; genetics ; DNA, Viral ; analysis ; Drug Resistance, Viral ; genetics ; Female ; Hepatitis B ; drug therapy ; virology ; Hepatitis B virus ; drug effects ; enzymology ; genetics ; Humans ; Lamivudine ; pharmacology ; therapeutic use ; Male ; Microbial Sensitivity Tests ; Middle Aged ; Point Mutation ; Polymerase Chain Reaction