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.

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.

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.

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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.

BACKGROUND:Microencapsulated cels are commonly used as a tool to overcome immune rejection after subarachnoid transplantation. However, the effect of microencapsulation on the secretion of human pheochromocytoma cels is unclear.
OBJECTIVE:To observe the growth and secretion of primarily microencapsulated cultured human pheochromocytoma cels in artificial cerebrospinal fluid.
METHODS: The human pheochromocytoma tissues were digested successively to isolate human pheochromocytoma cels that were then cultured in artificial cerebrospinal fluid. Primary cels were covered with alginate-polylysine-alginate microcapsules, and then the cel morphology was observed with inverted phase contrast microscope. Levels of met-enkephalin and norepinephrine in cel culture medium were detected by enzyme-labeled immunosorbent assay (ELISA). We used cel counting kit-8 colorimetric assay to obtain the growth curve of human pheochromocytoma cels in artificial cerebrospinal fluid.
RESULTS AND CONCLUSION:Microcapsulated human pheochromocytoma cels were in suspension and the process outgrowth increased slowly. Compared with non-microcapsulated cels, the proliferation rate of microcapsulated cels increased significantly. ELISA results revealed a significant increase in the levels of met-enkephalin and norepinephrine secreted from the microencapsulated cels compared to the non-microcapsule group. There was a wide variation in contents of met-enkephalin and norepinephrine from different tumors. These findings indicate that microencapsulated human pheochromocytoma cels can survive wel and have good secretion function in artificial cerebrospinal fluid, and human pheochromocytoma cels from different tumor tissues have stable secretory function.

Objective To summary and explore the indications,safety and effectiveness of no touch ostial cardioplegia delivery method during aortic valve replacement(AVR) for severe aortic insufficiency(AI),in order to provide guidance for clinical application.Methods Between September 2012 and February 2015,63 cases were randomly divided into two groups,in study group we used this technique in 31 patients with a mean age of(58.0 ± 15.0) years and a range of 31 to 74 years old.The degree of AI averaged (3.5 ± 0.5) according to a semiquantitative 5-grade scale[1].32 patients were enrolled in control group with a mean age of(60.4 ± 9.4) years and a range of 22 to 73 years old.The degree of AI averaged (3.6 ± 0.5).The abnormal aortic cusps were sutured though the aortic incision,which would be closed temporarily.Blood cardioplegia was infused repeatedly into the coronary osita through the closed aortic incision during the remainder of the procedure except AVR,when the closed aortic incision was reopened.Results In the study group,spontaneous resumption of nodal and normal sinus rhythm shortly after the removal of aortic cross-clamp was achieved in all patients,no hearts had to be electrically defibrillated and electrocardiogram showed no obvious S-T segment elevation.The time from cross-clamp removal to sinus rhythm was(12.3 ± 7.7) minutes in the study group,versus (18.6 ± 8.2) minutes in the control group(P =0.037).No operative mortality occurred in both groups.There was a significant increase in left ventricular ejection fraction[from(0.54 ± 0.09) preoperatively to(0.62 ± 0.06) postoperatively] (P =0.005) in patients of the study group,which was not seen in patients of the control group.Conclusion Preliminary results have shown that the no touch ostial cardioplegia delivery method is a feasible method for cardioplegia infusion during aortic valve replacement in patients with severe AI.