OBJECTIVETo study the antigenic properties of mutant hepatitis B virus surface antigen, to understand the sensitivity of the commercially available HBsAg assays to the variants and to reduce the undetectability of the variants.

METHODSRecombinant eukaryotic expression plasmids for HBsAg. The recombinant eukaryotic expression plasmids pSS1adr, pSS1adw2, pSS1adw2- 145Arg, pSS1adr-126 Asn and pSS1adr-126Ser were transfected into COS-7 cells. HBsAg in the supernatants of transfected cells was detected by using different commercial ELISA kits.

RESULTSThe absorbance value of pSS1adr-126 Asn and pSS1adr-126Ser plasmids were similar to that of the wild type HBsAg, the absorbance value of pSS1adw2-145Arg plasmids was lower than that of the wild type HBsAg.

CONCLUSIONIt is estimated that the antigenicity of HBsAg mainly depended on the amino acid sequence of "a" antigen determinant and its conformation, so 145 amino acid substitutions led to the change of conformation and the antigenicity of variant HBsAg was lower than that of the wild type.

Animals ; COS Cells ; Cercopithecus aethiops ; Culture Media, Conditioned ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Genetic Vectors ; genetics ; metabolism ; Hepatitis B Surface Antigens ; analysis ; genetics ; immunology ; Mutation ; Transfection ; Viral Envelope Proteins ; genetics

OBJECTIVETo determine the main genotype of hepatitis B virus (HBV) in Xinjiang.

METHODS200 HBsAg positive serum specimens were detected from more than 2000 serum of Xinjiang inhabitants, and HBV S gene was detected by using nPCR amplifying, and compared with the standard S region HBV nucleotide sequences of genotypes A-H retrieved from GenBank, then analyzed and drawn the polygenetic tree by MEGA3 software.

RESULTGene in 127 (63.5%) serum specimens was detected from 200 samples. Among 127 serum specimens, 10 (7.8%) was genotype B, 58 (45.7%) was genotype C, and 59 (46.5%) was genotype D.

CONCLUSIONGenotype B, C and D have been found in Xinjiang.

Adolescent ; Adult ; Aged ; Child ; China ; Female ; Genotype ; Hepatitis B ; ethnology ; virology ; Hepatitis B virus ; genetics ; isolation & purification ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Rural Health ; Young Adult

OBJECTIVETo construct full-length hepatitis B core particles presenting preS1 aa 21-47 epitope and truncated core particles presenting preS1 aa 37-45 epitope on their surface and compare their antigenicity.

METHODSPreS1 aa21-47 epitope and aa 37-45 epitope were inserted respectively into full-length hepatitis B core (aa 1-183) and truncated HBcAg (aa 1-144), between the 78th (Asp) and 79th (Pro). The genes synthesized after the codon optimization were ligated to the pET43. 1a vector with the same cohesive terminal (NdeI and XhoI) and expressed in the E. coli expression system. The morphology of the proteins of interest were observed by electron microscope and characterized by ELISA and Western Blotting.

RESULTSThe morphology of the virus-like particles were confirmed by electron microscope. H2 were solid particles with a diameter of (31.61 +/- 1.27) nm, while H3 were hollow particles with a diameter of (28.46 +/- 1.16) nm. Statistical analysis showed that H2 is larger than H3 in the diameter (P < 0.01). The antigenicity of the inserted epitopes and carrier protein were identified by ELISA and Western Blotting.

CONCLUSIONChimeric hepatitis B core particles presenting the preS1 neutralizing epitopes on their surface have been expressed, purified and identified, which lays the foundation for its application in vaccine research.

Epitopes ; chemistry ; genetics ; immunology ; Hepatitis B ; immunology ; virology ; Hepatitis B Core Antigens ; chemistry ; genetics ; immunology ; Hepatitis B Surface Antigens ; chemistry ; genetics ; immunology ; Hepatitis B virus ; chemistry ; genetics ; immunology ; Humans ; Neutralization Tests ; Protein Precursors ; chemistry ; genetics ; immunology ; Recombinant Fusion Proteins ; chemistry ; genetics ; immunology

Objective:To discuss the topical action characteristics of the biological transmission of moxibustion heat via temperature collection and numerical modeling.Methods:Temperature of moxibustion was measured at multiple points at a distance of 3 cm to obtain the moxibustion temperature field nephograms by the high-accuracy temperature measure array.Finite element analysis was used to imitate the three-dimensional dynamic distribution of temperature in acupoint tissues.Results:Through numerical analysis,the one-dimensional,two-dimensional and three-dimensional distributions of temperature in human acupoint tissues at 5 min of moxibustion were established.The result showed that moxibustion heat mainly transmitted from the surface of the tissue to the internal,and the influence of moxibustion heat decreased with the depth of the tissue.The analysis of the nephograms of acupoint tissue temperature at 5,10,15 and 20 min of moxibustion showed that with the increase of the moxibustion time,the temperature in acupoint tissues constantly rose,and the transmission depth of moxibustion heat also further expanded inside acupoint.Conclusion:By establishing the three-dimensional dynamic model of heat transmission inside acupoint tissues with the biological parameters of human tissues and the temperature values obtained,this study used finite element analysis software ANSYS 14.0 and discovered the rules in the transmission of heat in body tissues during moxibustion,and the features in moxibustion heat transmission (from the proximal to the distant) and heat penetration (from the surface to the internal).This study provides theoretical and experimental support for the application of moxibustion in clinical practice.

To explore new measures for functional reconstruction of multiple severe deformities as a result of extensive deep burn (total burn surface area > or = 90% TBSA, including deep burn > or = 70%TBSA) in late stage. Twelve severe burn patients with above-mentioned deformities were hospitalized in our ward during 1960--2005, the scars resulted from burns were distributed from head to foot with 173 deformities, including 27 scar ulcers. All patients lacked of self-care ability, among them some could not stand. Due to inadequate skin source, deformities were corrected by skin from matured scars expanded with subcutaneous balloon at late postburn stage. Following our former clinical experience, anatomic investigation and experimental research, we chose the following methods to correct deformities and restore functions: application of split-thickness scar skin after expansion (88 wounds); use of scar skin flap/scar-Achilles tendon flaps (59 wounds); combination of thin split-thickness skin grafts from scar and allogeneic acellular dermal matrix (composite skin, 40 wounds). All grafts survived, the appearance and function were improved obviously without complications. Follow-up 1-40 years, all patients could take care themselves with satisfactory function and appearance, and among them 8 patients returned to work (one had worked for 40 years), 2 patients married and had children. The above-mentioned measures are safe, reliable and effective for functional reconstruction of deformities.
Adult ; Burns ; complications ; surgery ; Cicatrix ; etiology ; surgery ; Contracture ; etiology ; surgery ; Female ; Humans ; Male ; Middle Aged ; Reconstructive Surgical Procedures ; methods ; Recovery of Function ; Skin Transplantation ; Skin, Artificial ; Surgical Flaps ; Wound Healing

OBJECTIVETo clone human PD-1 gene, construct a prokaryotic expression plasmid and express in E. coli.

METHODSThe human PD-1 cDNA was cloned by RT-PCR from the total RNA, which was extracted from peripheral blood lymphocyte cell of the patient with chronic hepatitis B. Recombinant PD-1 protein was been expressed and purified after the prokaryotic expression plasmid had been constructed. It was identified by SDS-PAGE, DNA sequencing and amino acid sequencing.

RESULTSThe PD-1 gene was cloned and confirmed by DNA sequencing. The recombinant protein was expressed in E. coli. The purified protein was obtained, then been confirmed by amino acid sequencing.

CONCLUSIONThe human PD-1 gene was successfully cloned and expressed in E. coli, which lays the foundation for further study on the function and application of PD-1.

Amino Acid Sequence ; Antigens, CD ; biosynthesis ; chemistry ; genetics ; isolation & purification ; Apoptosis Regulatory Proteins ; biosynthesis ; chemistry ; genetics ; isolation & purification ; Cloning, Molecular ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; genetics ; Genetic Vectors ; genetics ; metabolism ; Humans ; Polymerase Chain Reaction ; Programmed Cell Death 1 Receptor ; Prokaryotic Cells ; metabolism ; Sequence Alignment ; Sequence Analysis, DNA

OBJECTIVETo identify the role of p38 MAPK- NF-kB signaling pathway in TNF-α induced IL-8 production in human hepatocellular carcinoma cells.

METHODSThe concentrations of IL-8 from MHCC-97H cells were measured by an enzyme-linked immunosorbent assay (ELISA). The phosphorylation of p38 MAPK was analyzed by Western blot and immunofluorescence. NF-kB p65 protein nuclear translocation was determined by non-radioactive NF-kB p50 / p65 transcription factor activity kit and immunofluorescence.

RESULTSThe IL-8 production from MHCC-97H cells challenged with TNFa significantly increased in a time-dependent (F = 144.04, P < 0.01) and dose-dependent (F = 364.14, P < 0.01) manners, as compared with those without TNFa challenge. TNFa up-regulated the phosphorylation levels of p38 MAPK and increased the translocation of NF-kB p65 protein into the nucleus, also proved by immunofluorescence staining. p38 MAPK inhibitor (SB203580) could significantly inhibit IL-8 production in a dose-dependent manners (F = 65.47, P < 0.01), and partially inhibited NF-kB p65 nuclear translocation in a dose-dependent manner (F=141.20, P < 0.05).

CONCLUSIONTNF-α could increase the production of IL-8 in MHCC-97H cells and p38 MAPK- NF-kB pathways seem to play a central role in the regulation of IL-8 production.

Carcinoma, Hepatocellular ; metabolism ; Cell Line, Tumor ; Humans ; Interleukin-8 ; metabolism ; Liver Neoplasms ; metabolism ; Phosphorylation ; Signal Transduction ; Transcription Factor RelA ; metabolism ; Tumor Necrosis Factor-alpha ; pharmacology ; p38 Mitogen-Activated Protein Kinases ; metabolism

OBJECTIVETo evaluate the virological, serological and biochemical outcomes of 3 years of entecavir (ETV) treatment in nucleoside-naive chronic hepatitis B patients.

METHODSThis study was divided into two stages: Patients receiving either ETV 0.5 mg/d (n = 258) or lamivudine (LAM) 100 mg/d (n = 261) entered the initial 96-week randomized, double blind, controlled efficacy study. Patients not achieving a consolidated response (HBV DNA less than 0.7 MEq/ml, ALT less than 1.25 times*ULN, and if HBeAg-positive at baseline, loss of HBeAg for >or= 24 weeks), or those experienced viral breakthrough or relapse, entered a 48-week entecavir rollover study.

RESULTS96 weeks after the treatment, 79% of ETV treated and 46% of LAM treated patients had HBV DNA less than 300 copies/ml (P < 0.0001), 96% of ETV treated and 92% of LAM treated patients had normalized ALT (P = 0.06). 21% of ETV treated and 23% of LAM treated patients achieved HBeAg seroconversion. Among the 160 patients received continuous ETV for 144 weeks, 89% had undetectable serum HBV DNA, 86% showed ALT normalization, and 27% achieved HBeAg seroconversion. ETV resistance was rare: only 3 patients showed ETV resistance 96 weeks after the treatment, and additional 2 patients developed ETV resistance during the following 48 weeks, genotyping indicated the ETV resistance was caused by gene mutation. Adverse event rates in ETV-treated patients were similar to those in LAM-treated patients, but fewer ALT flares were observed in ETV-treated patients.

CONCLUSIONSThis study demonstrates that ETV treatment results in long-term HBV suppression and ALT normalization in Chinese CHB patients, and is associated with low rate of drug resistance.

Adolescent ; Adult ; Aged ; Alanine Transaminase ; blood ; Antiviral Agents ; administration & dosage ; therapeutic use ; DNA, Viral ; blood ; Double-Blind Method ; Drug Resistance, Viral ; Female ; Guanine ; administration & dosage ; analogs & derivatives ; therapeutic use ; Hepatitis B e Antigens ; blood ; Hepatitis B, Chronic ; blood ; drug therapy ; virology ; Humans ; Lamivudine ; administration & dosage ; therapeutic use ; Male ; Middle Aged ; Time Factors ; Treatment Outcome ; Viral Load ; Young Adult

OBJECTIVETo establish a rat model of scalding with controllable depth and area by high pressure steam.

METHODSHigh pressure steam apparatus consisting of an autoclave and a self-made scalding frame was employed in the study. The rats were inflicted with scalding with 0.12 Mpa (1 Mpa = 7500 mmHg) high pressure steam on the back through a hole of 2.6 cm in diameter for 3, 4, 5, 6, 7, 8, 9 and 10 seconds, with five wounds at each time point. The tissue samples were harvested at 24 post injury hour (PIH) for pathomorphological examination. The depth of scald was measured, and injury to the sweat gland and hair follicles injury, the hair growth in scalded area, and the wound healing condition were observed with Photoshop software.

RESULTSThere was positive correlation between the scalding depth and scalding time. The injury time for superficial and deep partial thickness burn and full thickness burn were 3, 5 and 7 seconds respectively. The wound healing time was similar even the scalding became more and more serious when injury time increased from 7 to 10 seconds.

CONCLUSIONThe scalding depth and area in this model could be controlled, and the degree of scald injury could be graded accurately with easy manipulation. The result showed that it was an ideal model of skin burn wound.

Animals ; Burns ; pathology ; Disease Models, Animal ; Male ; Pressure ; Rats ; Rats, Sprague-Dawley ; Steam

OBJECTIVETo explore new methods to repair postburn contracture deformities in patients with extremely limited donor skin.

METHODSFive severely burned patients with extremely limited donor skin but severe deformities were enrolled in the study. The mature and the pliable scarred skin was utilized as the donor site for reconstruction of the postburn deformities. Split-thickness scarred skin was harvested for repair of postburn deformities after the scarred skin was expanded by expander, while thin razor-thin scarred skin with allo-acellular dermal matrix (ADM) was employed for the repair of postburn deformities when skin expansion was not feasible.

RESULTSAll the expanded scarred skin and composite skin grafts survived completely with good function and configuration. The long-term follow-up result was satisfactory, and the grafted skin was similar to that with split-thickness skin grafting.

CONCLUSIONIt is feasible to employ various split-thickness scarred skin for the reconstruction of postburn deformities. This technique is a new effective procedure for the reconstruction of postburn deformities, especially for those with extreme scarcity of donor site.

Adult ; Burns ; pathology ; surgery ; Cicatrix ; pathology ; Contracture ; surgery ; Female ; Humans ; Male ; Middle Aged ; Skin Transplantation ; methods