Objective To explore the clonality of the T cells and the role of cellular immunological pathogenesis in chronic asymptomatic HBV carriers (AsC) by TCR CDR3 size spectratyping and determining sequence. Methods The TCR CDR3 region genes of 24 BV families were amplified by utilizing inverse polymerase chain reaction (RT-PCR) technology, and the CDR3 size lengths of T cell receptor (TCR) ?-chain were analyzed with Genescan technology for 4 healthy individuals and 9 AsCs. The clonality of T cells presumed by spectratyping was further confirmed by CDR3 sequencing. Results The CDR3 repertoire of 4 healthy individuals showed Gaussian distribution. The clonal expansions of T cell were observed in 8 out of 9 AsCs. The expanded T cells have different CDR3 sequences. Conclusion There is significantly clonal expansion in the compartment of the peripheral blood T lymphocyte in AsCs. The expanded T cells do not have homogenicity.

Objective To explore the formation process of vacuum pressure differential of micro power negative pressure technology to facilitate its clinical application.MethodsThe formation of negative pressure differential and changes of negative pressure values in enclosure space were studied which was formed by liquid aspiration after the compressive polyvinyl alcohol foam material was enclosed.Micro power negative pressure technology was applied to the treatment of the right ear necrosis patient after focal cleaning,and the efficacy was observed.Results Medical adhesive film was raised gradually during the expansion of polyvinyl alcohol foam due to liquid aspiration,and then the enclosure space extended increasingly to form local vacuum negative pressure.The range of maximal negative pressure was between-110 and-200 Pa,and the mean maximal negative pressure value was-132.7 Pa.Micro power negative pressure technology behaved well in healing the wound.Conclusion Micro power negative pressure technology gains advantages in reliability and easy operation over other technologies and products,and may be a new choice for healing of refractory wounds.

To investigate the mechanism of binding of human serum albumin (HSA) with potential sensitinogen, including chlorogenic acid and two isochlorogenic acids (3,4-di-O-caffeoylquinic acid and 3,5-di-O-caffeoylquinic acid).

0.05).(3) IL-17 was positively correlated to CRP(r=0.460,P 0.05).Conclusion:(1) IL-17 in patients is much higher than that in normal people,and is positively correlated to CRP,the acknowledged marker of inflammation status,but not correlated to dialysis duration,suggesting that IL-17 plays a role in pathogenesis of immune anomaly and micro-inflammation in patients with ESRD and HD;(2) There is no correlation between IL-6 and IL-17,and the mechanism of IL-17 elevation in ESRD patients needs to be further investigated.

Objective To investigate a new microcapsular system for culture of primary generation hepatocytes in vitro. Methods A novel microcapsular system was used for in vitro culture of primary generation rat hepatocytes .The various characteristics of the microcapsular system were examined. The survival rate of hepatocytes and the functions of urea and albumin synthesis were observed.Results The microcapsule had good permeability,a certain degree of mechanical stability and favorable immunological protective ability. Hepatocytes within the microcapsule maintained a high degree of viability and function, and the synthesis of urea and albumin was 1~3 times higher than that of controls.Conclusions This microcapsule has excellent biological and mechanical properties.This system can be of great benefit for use in bioartificial liver and cell transplantation.

Objective:To acquire potential HBsAb sequences,we have analyzed the BCR CDR3 repertoire of the peripheral blood with HBsAb titer higher than 10 000 mU/ml,which could provide a data basis for follow-up study.Methods:Genomic DNA of pe-ripheral blood mononuclear cells was extracted from samples with HBsAb titer higher than 10 000 mU/ml.We have adopted Illumina Solexa high-throughput sequencing technology of the Adaptive Biotechnologies ImmunoSEQ platform to acquire sequence data.IMGT/High V-QUEST was used to preliminary analyze our sequence data,including usage of IGHV,IGHJ and IGHD gene subgroups,IGHV-J matching,distribution of CDR3 amino acid (AA) length and usage of total CDR3 AA.And these sequences were compared with the HBsAb sequences from NCBI database.Results:Experimental samples have highly selected gene subgroups IGHV3,IGHV4,IGHJ4, IGHJ6,IGHD3,IGHD6,and IGHV3-J4 pairing,IGHV3-J6 pairing.The AA length distributions of CDR3 region were normal distribution with the length of 14/15 AA as the midline.In the regard to amino acid usage in CDR3 region, each sample prior used Alanine, Tyrosine,Glycine,Alanine,Aspartic acid and Serine.The amino acid usages of 107,108,109,113,114 positions were diversified but 105,106,115,116,117 positions taking conservative amino acids usages.We have found 48 unique sequences that have same IGHV, IGHJ and CDR3 AA length with the HBsAb sequences from NCBI database.Conclusion:There were almost the same characteristics of BCR CDR3 repertoire of the peripheral blood with HBsAb titer higher than 10 000 mU/ml.The 48 unique sequences provided a solid data basis for the follow-up study.

CD4-Positive T-Lymphocytes ; Hepatitis B virus ; Hepatitis B, Chronic ; Humans ; Interleukin-2 Receptor alpha Subunit ; Sequence Analysis

Objective To analyse the spectral patterns of complementarity determining region 3 (CDR3) length distribution of T lymphocyte receptor beta chain variable (TRBV) gene families in infiltrating T cells of the liver tissues and the peripheral blood samples of patients with chronic hepatitis B (CHB) in order to evaluate the characteristics of T cell clonal expansion. Methods The spectral patterns drift of TRBV gene families (the monoclonal/oligoclonal TCR β T cells) in the peripheral blood and hepatic tissues from 11 cases of CHB patients were analyzed by the real-time fluorescence quantitative reverse transcription polymerase chain reaction (FQ-PCR) with DNA melting curve analysis, and abnormal rates of TRBV gene families were compared between CHB patients and healthy control. The comparison of rates was done by chi square test. Results The gene melting spectral pattern of 26 TRBV families of the 11 CHB patients, no matter in the peripheral blood or hepatic tissue, showed either a single peak or prominent melting peaks, even disappeared for certain TRBV families. The abnormal rate of TRBV gene families in the hepatic tissues was significantly higher than that in the peripheral blood samples (x2 = 23. 246, P＜0. 01). What is more interesting was that some parts of TRBV families were identical in both the peripheral blood and the hepatic tissue in certain patients. TCR BV13.1, TCR BV17 and TCR BV22 fragments were found to be restricted used in both the peripheral blood and hepatic tissue by some CHB patients. Conclusions T cells in the peripheral blood and the hepatic tissues of CHB patients can develop clonal expansion to some extent.Parts of TRBV families are restricted used in the peripheral blood and hepatic tissue in some CHB patients, which offers a foundation for further studying the common specific spectral drift patterns of TRBV CDR3 gene in CHB patients.

Objective To investigate the effect of postremission consolidation therapy with intermedium-dose cytarabine (MDAC) in elderly patients with acute myelogenous leukemia (AML). Methods Clinical data of 61 elderly AML patients (except M3) in postremission who achieved complete remission (CR) in two period of remission induction program were retrospectively analyzed. Results There were 26 cases in MDAC group and 35 cases in standard-dose cytarabine (SDAC) group. In MDAC group and SDAC group, the relapse free survival (RFS) time were 42.7 months and 16.0 months respectively (P= 0.002), the overall survival (OS) time were 44.6 months and 18.2 months respectively (P= 0.004), and the cumulative relapse frequencies rates were 26.9 % (7/26) and 54.3 % (19/35) respectively (x 2= 4.567, P= 0.033). However, 3 years OS rate of the two groups were 23.1%(6/26) and 8.6%(3/35) (x 2=2.496, P=0.114) , and there was no significant difference in the incidence of adverse reactions between the two groups (all P > 0.05). Conclusion MDAC could improve RFS and OS for the elderly AML patients in postremission who received CR in the early stage, and the incidence of adverse reactions is similar to that of SDAC.

Objective:To establish the technique for real-time fluorescence quantitative reverse transcription polymerase chain reaction(FQ-PCR)by DNA melting curve analysis for detecting the CDR3 shewing of TCR alpha gene repertoire in human peripheral blood.Methods:Total RNA of peripheral blood mononuclear cell(PBMC)from 4 healthy donors and 2 patients with lymphomatous leukemia were transcripted reversely into cDNA.The cDNA of 32 TRAV gene family CDR3 was amplified by FQ-PCR.Analysis of the monoclonal/oligoclonal/polyclonal CDR3 spectratyping with DNA melting curve.Results:The FQ-PCR products of 32 TRAV family CDR3 were showedas a blur land at the predicted of products size in healthy donors and parts of TRAV family CDR3 products disappeared in patients on 1.5% agarose gel by Gold-View staining.The 32 TRAV family CDR3 were showed with different frequencies by relative fluorescence quantitative in healthy donors and the patients.The CDR3 spetratyping for 32 TRAV families was showed as polyclonal peak(Gaussian distribution)in healthy donors but showed as different monoclonal/oligoclonal/polyclonal peak in the patients with lymphomatous leukemia with DNA melting curve analysis(we called "melting curve spectratyping of CDR3")Conclusion:The study suggests that the technique of "FQ-PCR with DNA melting curve analysis be convenience and celerity for detecting the CDR3 skewing of TCR alpha gene repertoire in human peripheral blood.