Objective:To prepare and test tetrameric sH-2K~d-HBc complex for the further measurement of the specific CTL response.Methods:PE labled streptavidin with 4 biotinylated binding sites can bind to 4 biotinylated monomer to form the corresponding tetramer.Mice were immunized via different methods of genetic immunization by use of the construted pcDNA3-C plasmid to get the specific CTLs.Then our prepared tetramer was applied to stain the specific CTLs by the analysis of flow cytometry.Results:We applied our prepared tetramer to stain the cells from the experimental groups and control group.The results showed the tetramer was able to discriminate the frequencies of specific CTL induced by the three immunol methods(0.24%,0.26%,0.36% vs 0.07%,P≤0.05).This demonstrated that the prepared tetramer could bind its targets specifically and efficiently.The three immunol methods induced different levels of immune responses.Compared with the traditional muscle injection,gene gun induced weaker humoral immune response and stronger cellular immune response,and hydrodynamic injection induced the strongest humoral and cellular immune responses.Conclusion:Have successfully constructed the sH-2K~d-HBc tetramer.The techniques and methods can be used for preparation of tetramers of other types of MHCⅠ molecules.

Objective To investigate the role of renal nerve in cardioprotection provided by renal ischemic preconditioning(RIP).Methods The effects of ischemia-reperfusion and RIP on the hemodynamics, myocardial oxygen consumption, epicardial electrography and infarct size were examined in anesthetized rabbit.Results During the 45 min of myocardial ischemia and 180 min of reperfusion, all hemodynamic parameters and myocardial oxygen consumption decreased progressively significantly. Epicardial electrographic ST-segment was elevated significantly during myocardial ischemia and return to baseline progressively in the course of reperfusion. The myocardial infarct size occupied 55.80±1.25% of area at risk,and RIP significantly reduced the myocardial infarct size to 36.51±2.80%(P＜0.01). The renal nerve section (RNS) per se didn't affect myocardial infarct size produced by ischemia-reperfusion, while cardioprotection afforded by RIP was completely abolished by RNS.Conlusion RIP have the protective effect on heart, and activation of renal afferents by transient ischemia-reperfusion play an important role in such a cardioprotection.

OBJECTIVE:To reduce the risk of single dispensing,and to improve the quality of pharmaceutical care in emergen-cy pharmacy. METHODS:Single dispensing environmental factors,human factors and social factors of emergency pharmacy in our hospital were analyzed;the suggestions and measures were put forward,risk intervention mechanism was established and manage-ment effects were evaluated. RESULTS:Through improving the pharmacy hardware and software,carrying out the relevant sys-tem,enhancing training,improving the process of dispensing and window order,promoting the drug management and carrying out near error management. Compared with before management(Oct.-Dec. 2014),the times of near error and disputes significantly re-duced after management(Feb.-Apr. 2015)(P<0.05),and medication error reduced to zero. CONCLUSIONS:As single dispens-ing,risk management and the improvement of prescription audit ability can improve pharmaceutical care in emergency pharmacy.

This study investigated the expression profiles of IL-10 gene in three human hepatoma cell lines including Huh7, HepG2, and HepG2 transfected with a plasmid containing hepatitis B virus (HBV) named HepG2.2.15. RT-PCR analysis demonstrated that IL-10 message RNA was absent in HepG2 and Huh7 cells, whereas it was present in HepG2.2.15 cells, which was consistent with ELISA result. Furthermore, except for lamivudine other antiviral treatments did not significantly decrease the HBV DNA level in HepG2.2.15 cells, while they had different effects on the expression of IL-10 protein, although stimulation by LPS had no significant effect. In addition, except for poly(I:C), the other treatments decreased the expression of IL-10 protein to different degrees, but had no significant effects on the expression of NF-κB and MyD88. Meanwhile, all treatments we used had effect on the expression of STAT1. In conclusion, IL-10 was expressed in HepG2.2.15 cells and STAT1 pathway might be involved in the regulation of IL-10 expression in HepG2.2.15 cells, but it was not the sole pathway, the exact mechanism warrants further study.

Adenocarcinoma ; metabolism ; pathology ; Adenocarcinoma, Follicular ; metabolism ; pathology ; surgery ; Bronchial Neoplasms ; metabolism ; secondary ; surgery ; Carcinoid Tumor ; metabolism ; pathology ; DNA-Binding Proteins ; metabolism ; Diagnosis, Differential ; Female ; Humans ; Middle Aged ; Thyroglobulin ; metabolism ; Thyroid Neoplasms ; metabolism ; pathology ; surgery ; Transcription Factors

Objective To investigate the effects of antioxidant SS-31 on sepsis-induced acute lung injury (ALI)in a mouse model of sepsis.Methods Sepsis was induced in male mice by cecal liga-tion and puncture (CLP).Forty-eight adult male mice (C57BL/6,weight 25-32 g)were equally as-signed to the sham+vehicle group (group A),sham+SS-31 group (group B),CLP+vehicle group (group C),or the CLP+SS-31 group (group D).At 0 or 5 h after CLP or sham operation,mice re-ceived an intraperitoneal injection of SS-31 (5 mg/kg of body weight)or the same volume of normal saline.Pulmonary tumor necrosis factor alpha (TNF-α),interleukin (IL)-6,IL-10,malondialdehyde (MDA),superoxide dismutase activities (SOD),myeloperoxidase activities (MPO ),wet-to-dry weight ratio (W/D),reactive oxygen species (ROS),ATP,NF-κB p65,inducible nitric oxide syn-thase (iNOS),and histological scores were assessed 24 h after operation.Results Pneumonia,edema were significantly heavier in group C than in group A (P <0.05).Lung congestion,inflammatory cell infiltration,alveolar wall edema was significantly less in group D than in group C (P <0.05).Pulmo-nary histological scores,IL-6,MDA,MPO,W/D,ROS,NF-κB p65 and iNOS significantly in-creased,while ATP levels decreased in group C when compared with group A (P <0.05).However, SS-31 treatment significantly reversed these parameters when compared with the group C (P <0.05). No difference was observed between the group A and group B.There was no difference of TNF-α,IL-10,and SOD among the four groups.Conclusion SS-31 improves sepsis-induced ALI in a mouse model probably by down-regulating the oxidative stress and inflammation in of sepsis-induced ALI.

Objective To explore the correlation of traditional Chinese medical syndrome elements with plasma connective tissue growth factor ( CTGF) and platelet-derived growth factor ( PDGF) in early liver cirrhosis induced by type B hepatitis. Methods The distribution of traditional Chinese medical syndrome elements in early liver cirrhosis induced by type B hepatitis was analyzed, plasma contents of CTGF and PDGF were detected by enzyme-linked immunosorbent assay ( ELISA) , and the correlation of syndrome elements with CTGF and PDGF was discriminated. Results ( 1) The distribution of traditional Chinese medical syndrome elements in early liver cirrhosis induced by type B hepatitis showed as follows: the syndrome elements involved the viscera of liver and spleen, and the pathogenesis was characterized as dampness, heat, qi stagnation, and yin deficiency. ( 2) CTGF was closely related with spleen, gallbladder and dampness, with OR value being 1.598, 1.567, 2.797, respectively. PDGF was closely related with heat, with OR value being 1.134. Conclusion Early liver cirrhosis induced by type B hepatitis mainly affects the viscera of liver and spleen, the pathogenesis is characterized by dampness, heat, qi stagnation, and yin deficiency. The patients with higher CTGF are apt to show the pathological changes of spleen, gallbladder, dampness, and have the syndrome el-ements of spleen, gallbladder, dampness. The patients with higher PDGF are apt to show the pathological changes of heat, and have the syndrome element of heat.

Hepatitis B virus (HBV)-specific cytotoxic T lymphocytes (CTLs) are believed to play a major role in viral clearance and disease pathogenesis during HBV infection. To clarify the differences in host immune responses between self-limited and chronic HBV infections, we constructed three HLA-A*0201/HBV tetramers with immunodominant epitopes of core18-27, polymerase 575-583 and envelope 335-343, and analyzed the HBV-specific CTLs in peripheral blood mononuclear cells (PBMCs) from patients infected with HBV. The frequencies and expansion ability of HBV-specific CD8(+) T cells in most self-limited HBV infected individuals were higher than those in chronic HBV-infected patients. HBV-specific CD8(+) T cells could be induced by in vitro peptide stimulation from chronic patients with a low level of serum HBV-DNA but not from those with a high level of serum HBV-DNA. In chronic infection, no significant correlation was found either between the frequencies of HBV-specific CD8(+) T cells and the viral load, or between the frequencies and the levels of alanine transaminase. Our results suggested that the frequencies of HBV-specific CTLs are not the main determinant of immune-mediated protection in chronic HBV infection and immunotherapeutic approaches should be aimed at not only boosting a HBV-specific CD8(+) T response but also improving its function.

AIM: To prepare and identify mouse polyclonal antibody against protein Hlb, which is the variant of major subunit of human ASGPR. METHODS: Hlb specific peptide was synthesized and coupled with keyhole limpet hemocyanin (KLH) for immunization. Then H1b-KLH conjugation was injected into mouse subcutaneously to produce polyclonal antibody. ELISA assay was used to detect the titer of the antibody. Antibody was also identified by Western blot and immunohistochemistry assays. RESULTS: Mouse antibody against Hlb was prepared after injection of H1bKLH conjugation. The titer of H1b antibody was about 1:10~5.Western blot confirmed its high specificity. This antibody could also be used for immunohistochemistry analysis. CONCLUSION: The successful preparation of the polyclonal antibody against protein H1b, which can discriminate the two variants of the major subunit of ASGPR with high specificity, will provide an efficient reagent for further study of the physiologic functions of H1b and its role in the pathogenesis of human disease.

Objective To evaluate the effect of cardiovascular disease (CVD) on the healthcare costs associated with chronic kidney disease (CKD).Methods A total of 96 patients with stage 3-4 CKD treated at CKD clinic of Beijing Friendship Hospital,Capital Medical University were enrolled in the study.Their mean age was (61.6±9.5) years including 65 male and 31 female patients.A retrospective analysis of clinical material and health claims between January 2010 and October 2010 was conducted.Firstly,patients were grouped according to median CKD-associated healthcare cost and clinical characteristics were compared between two groups.Secondly,patients were stratified into three categories based on CVD prevalence (with acute cardiovascular events,with CVD but no acute events,and without CVD),and CKD-associated healthcare costs were assessed among the groups.Finally,the potential factors influencing CKD-associated healthcare costs were evaluated by optimal scaling regression analysis.Results During January to October in 2010,median CKD-associated healthcare costs was 13960.5 yuan (interquartile range 10226.5,19667.2 yuan).In the group with higher healthcare costs,more females,greater urine albumin-creatinine ratio,more emergency observations and/or hospitalizations caused by acute cardiovascular events,higher diabetes mellitus prevalence and calcium-phosphorus products,and lower eGFR and hemoglobin levels were found (P＜0.05,respectively).In contrast,the total prevalence of CVD was not significantly different between the groups (P=0.386).When grouping by CVD prevalence,significant differences of CKD-associated healthcare costs were observed only between patients with acute cardiovascular events and the other two groups (P＜0.01,respectively).The median healthcare cost of the former was approximately twice as higher as that of the other two groups,and the maximal cost was also found in the acute-cardiovascular-event group.For the optimal scaling regression analysis,both emergency observations and/or hospitalizations caused by acute cardiovascular events and diabetes mellitus entered the equation,and standardized coefficients were -0.538 and -0.217 respectively (P＜0.01 and P＜0.05).Conclusions Emergency observations and/or hospitalizations caused by acute cardiovascular events are important factors inducing high CKD-associated healthcare costs in patients with stage 3-4 CKD.Therefore,the prevention of acute cardiovascular events may be favorable to reduce CKD-associated healthcare costs.Larger and longer-time perspective studies are required to confirm it.In addition,diabetes mellitus also influences CKD-associated healthcare costs.