Objective Using the technology of siRNA to inhibit gene expression of T cells'nonreceptor tyrosine protein kinase Lck in asthmatic mice,and to study the effect of siRNA inhibited Lck to the function of T cells in asthmatic mice.Methods The 21 - 23 bp RNA fragments of mouse T cell Lck were made by chemosynthesis.INTERFERinTMsiRNA Transfection Reagent was used as transfection reagent to transfect the siRNA into the spleen T cells of asthmatic mice for 48 hours.Then T cells were mixed with bone marrow dendritic cells (DC) of asthmatic mice for another 48 hours.Cell culture suspension was collected and the level of IL-4,IL-13,IL-2,INF-γ were detected with respondent ELISA kits; Western Blot was used to identify if the expression of Lck was blocked.Results The expression of Lck in T cells almost could not be detected in siRNA interference group.The levels of IL-4 and IL-13 in siRNA interference group( 10.19 ± 1.66,12.34 ±0.79) were lower than no-siRNA interference(28.06 ±2.88,27.87 ± 1.61 )and control group ( 22.07 ± 2.5 1,20.47 ± 2.37 ),and the difference was statistical significant ( P ＜0.01 ).Conclusions Special siRNA could block the expression of special gene,and Lck specific siRNA could block the activation and differentiation of T cells and reduce the secretion of inflammatory cytokines in asthmatic mice.

Objective To study the gene expression profiles of megakaryocytes(MKs) from human cord blood CD34+ cells in vitro expanded and to understand megakaryopoiesis at the molecular level. Methods CD34+ cells were isolated using density gradient centrifugation and magnetic activated cell sorting. The cells were cultured and stimulated with recombinant human TPO ( 100 ng/ml). After 12 days, the MKs fraction was separated using an anti-CD41 monoclonal antibody by immunomagnetic sorting. The gene expression profiles of MKs, non-MKs as well as meg-01 cells were studied by gene chip assay. THBSI, HOX A9,β-actin, lL-8,Annexin A6, FGF-8 were selected to validate the gene chip results by RT-PCR. Results A total of 116 genes between MKs and non-MKs cells were significantly different, 52 genes were up-regulated and 64 genes were down-regulated. In addition, 158 genes between MKs and meg-01 cells were significantly different, 71 genes were up-regulated and 87 genes were down-regulated. THBSI showed higher expression in MKs than in non-MKs. HOXA9 showed lower expression in MKs than in non-MKs. The expression of β-actin did not show any significant difference in MKs and non-MKs. IL-8 showed higher expression in MKs than in meg-01 cells, while ANXA6 showed lower expression in MKs than in meg-01 cells. The expression of FGF-8 did not show any significant difference between MKs and meg-01 cells. Conclusions MKs, non-MKs and meg-01 cells show different gene expression profiles. The regulatory genes include stress response genes,immune related genes, DNA synthesis and repair genes, metabolism genes, pro-onco genes and tumor suppressor genes.

ObjectiveUsing the technology of siRNA to inhibit the gene expression of no-receptor　tyrosine protein kinase Lck in T cells of asthmatic mice,and to study the therapeutic effect of Lck specific　siRNA in asthmatic mice.MethodsReceptor tyrosine protein kinase Lck specific siRNA fragments were　taken from chemosynthesis.In vivo-jetPEITM was used to transfect the siRNA into mice body through tail　vein injection.The mice were killed 48 hours later,and the levels of IL-4,IL-17 in bronchoalveolar lavage　fluid (BALF) were detected with respondent ELISA kits.The change of inflammatory histopathology in lung　was observed with H.E.staining.The expression of Lck in lung was detected with immunohistochemistry　(IHC),and the level of Lck in lung tissue homogenate was detected with Western Blot.Results Compared with asthmatic group[ (234.68 ± 11.15 ) pg/ml,( 96.76 ± 8.28 ) pg/ml],the levels of IL-4,IL-17　[ (234.68 ± 11.15)pg/ml,(96.76 ±8.28) pg/ml] in the BALF of siRNA interference group decreased,　and the inflammation in the lung relieved.IHC indicated that the expression of Lck in lung decreased and　the level of Lck in lung tissue homogenate decreased ( P ＜ 0.05 ).Conclusions Lck specific siRNA　could reduce the level of IL-4,IL-17 in the lung tissues of asthmatic mice,and relieve the inflammatory reaction in lung.

Objective To recognize the CT manifestations of benign metastasizing leiomyoma (BML).Methods Four cases of BML proved by pathology were retrospectively analysed.Among these, 2 cases were pulmonary BML,1 ease was leiomyomatosis peritonealis diseminata(LPD),and 1 case was combined with pulmonary BML and LPD.All of them were female,and had a history of hysterectomy,the average age were 45.The CT findings of them were analyzed.Results The CT of pulmonary BML in three cases showed multiple rounded,well-defined discrete nodules,0.2—2.0cm in diameter,random distribution,and no calcification in the nodules.No hilum or mediastinum lymphadenopathy,and no pleural effusion.There were no enhancement after intravenous contrast examination.The CT findings of LPD in two cases were many masses around the vena cava,abdominal aorta and/or iliac artery.The masses were 4— 5cm in diameter with smooth margin and homogeneous density.No lymph node or ascites were found.One case who got intravenous contrast enhanced CT scan showed significant enhancement of the lesions;another case who got PET CT study showed no significant tracer concentration.Conclusion When multiple metastasizing nodular lesions with sharp margin and homogeneous density were found in the lungs or abdomen,they should be considered the possibility of BML in women with hysterectomy history.

BACKGROUND: Greatly importance has been attached to ceramic-on-ceramic bearing surface due to its excel ent wear resistance. But the risks of squeaking and ceramic fracture also go with it. Up til now, the choice between ceramic-on-ceramic and ceramic-on-polyethylene bearing surfaces in primary total hip arthroplasty remains controversial. OBJECTIVE: To compare the clinical outcomes and safety between ceramic-on-ceramic versus ceramic-on-polyethylene bearing surfaces in total hip arthroplasty based on meta analysis. METHODS: We electronical y searched databases including PubMed/Medline, Embase, Web of Science, Cochrane Col aboration database, Chinese Biomedical Literature Database (CBMdisc) and China National Knowledge Internet for randomized control ed trials on the comparison between ceramic-on-ceramic versus ceramic-on-polyethylene bearing surfaces in total hip arthroplasty from inception to January 2015. References of the included studies were also retrieved. Investigators severely selected the studies, extracted data and assessed the quality according to the inclusion and exclusion criteria. Then, meta-analysis was performed using RevMan 5.2 software. RESULTS AND CONCLUSION: Nine randomized control ed trials were included, involving 1 231 hips with ceramic-on-ceramic prosthesis and 932 hips with ceramic-on-polyethylene prosthesis. Meta analysis showed that both bearing surfaces achieved satisfied function recovery. But ceramic-on-ceramic had significantly increased risks of squeaking and ceramic fracture, meanwhile ceramic-on-polyethylene showed significantly higher wear rate. There were no significant differences in intra- or post-operative dislocation, osteolysis and other complications and prosthesis failure with any reason between two bearing surfaces. These results suggest that during the short- to mid-term fol ow-up period, no sufficient evidence can tel that ceramic-on-ceramic was obviously super than ceramic-on-polyethylene. Long-term fol ow-up is required for further evaluation.

AIMTo assess the parameters of cardiac structure and function of male Balb/c mice by the echocardiography.

METHODSA total of 27 male Balb/c mice (from five to seven week old) were examined with a 13-MHz transthoracic linear-array transducer, hearts were removed from mice anesthetized with Nembutal, and the left ventricular (LV) mass were weighed.

RESULTSComplete 2-dimensional echocardiography for cardiac structure and function were obtained. Hemodynamic parameters were recorded. A correlation existed between LV weight (x) and echocardiographic LV mass (y) with the 2D) guided M-mode method: y = 1.15x + 3.26, (r = 0.80).

CONCLUSIONEchocardiography appears to be a promising approach for noninvasively assessing LV mass and function in mice.

Animals ; Echocardiography ; Heart ; physiology ; Heart Ventricles ; diagnostic imaging ; Male ; Mice ; Mice, Inbred BALB C ; Ventricular Function, Left

OBJECTIVETo investigate the effect of immune modulation therapy on cardiac function and lymphocyte subsets in aged patients with chronic heart failure (CHF).

METHODSCHF (NYHA classification: II-IV) patients older than 60 years were randomly divided into two groups: CHF intervention group received regular therapy and thymopetide (2 mg/day i.m. for 75 days, n = 48), CHF control group received regular therapy (n = 48), 45 healthy individuals older than 60 years served as normal control. Left ventricular ejection faction of (LVEF), inner diameter of left ventricular end-diastole (LVEDD), inner diameter of left ventricular end-systole (LVESD), lymphocyte subsets, plasma high sensitive C-reactive protein (hsCRP), plasma brain natrium peptide (BNP) and 6 minutes walking distance (6MWT) were measured at before therapy, after the first course (15 days) of treatment and after the third course of treatment (75 days).

RESULTS(1) Before therapy, the levels of BNP, hsCRP, CD8 T cells, LVEDD and LVESD were significantly higher and the levels of CD3, CD4, CD19 T cells, NK, CD4/CD8 ratio, LVEF and 6MWT were significantly lower in CHF patients compared to compared normal controls (all P < 0.05). These parameters were similar between CHF intervention group and CHF control group. (2) At 15 days, the levels of CD3, CD4, CD19 T cells and NK were significantly increased (P < 0.05 or P < 0.01) while the level of CD8, BNP and hsCRP were significantly decreased (P < 0.05 or P < 0.01) in CHF intervention group compared with CHF control group. (3) At 75 days, the levels of CD3, CD4, CD19 T cells, NK, CD4/CD8, LVEF and 6MWT were significantly increased (P < 0.05 or P < 0.01) while the levels of CD8, BNP, hsCRP and Minnesota Living with Heart Failure Questionnaire (MLHFQ) were significantly decreased (P < 0.05 or P < 0.01) in CHF intervention group compared with CHF control group.

CONCLUSIONThymopetide, an immune modulating agent, might regulate the quantity and proportion of lymphocyte subsets and improve cardiac function in aged patients with CHF, indicating that immune modulation therapy might be a new treatment strategy for aged CHF patients.

Aged ; Aged, 80 and over ; C-Reactive Protein ; analysis ; Case-Control Studies ; Chronic Disease ; Female ; Heart Failure ; drug therapy ; immunology ; physiopathology ; Humans ; Immunologic Factors ; therapeutic use ; Lymphocyte Subsets ; Male ; Middle Aged ; Thymosin ; therapeutic use ; Ventricular Function, Left

Animals ; History, 20th Century ; Humans ; Influenza A Virus, H1N1 Subtype ; genetics ; immunology ; isolation & purification ; Influenza, Human ; epidemiology ; history ; immunology ; virology ; Russia ; epidemiology

Animals ; Global Health ; History, 20th Century ; Humans ; Influenza A Virus, H1N1 Subtype ; genetics ; isolation & purification ; metabolism ; pathogenicity ; Influenza, Human ; epidemiology ; history ; virology ; Spain ; epidemiology ; Viral Proteins ; genetics ; metabolism

OBJECTIVEReceptor activity-modifying proteins (RAMPs) determine the ligand specificity of the calcitonin receptor-like receptor (CRLR); co-expression of RAMP1 and CRLR results in a calcitonin gene related peptide (CGRP) receptor, whereas the association of RAMP2 or RAMP3 with CRLR gives an adrenomedullin(ADM) receptor. As CGRP and ADM may play a beneficial role in heart failure, this study aimed at the question whether RAMPs mRNAs are changed in heart failure.

METHODSSemi-quantitative reverse transcription-PCR (RT-PCR) was used to detect and quantify the mRNAs of RAMP1 and RAMP3 in the atria of heart failing patients.

RESULTSIt was found that the expressions of RAMP1, RAMP2 and RAMP3 mRNAs increased with the worsening of heart function, but the expressions of RAMP1 and RAMP2 mRNA decreased at level IV of heart failure.

CONCLUSIONThe above results demonstrated in the atria of heart failure patients an up-regulation of CGRP receptor by an increase of RAMP1 in association with CRLR and an up-regulation of ADM receptor by an increase of RAMP2 expression in association with CRLR, thus suggesting that CGRP and ADM receptors be playing a functional role in compensating the chronic heart failure in human.

Adult ; Calcitonin Receptor-Like Protein ; Female ; Heart Atria ; metabolism ; Heart Failure ; genetics ; physiopathology ; Humans ; Intracellular Signaling Peptides and Proteins ; genetics ; physiology ; Male ; Membrane Proteins ; genetics ; physiology ; Receptor Activity-Modifying Protein 1 ; Receptor Activity-Modifying Protein 2 ; Receptor Activity-Modifying Protein 3 ; Receptor Activity-Modifying Proteins ; Receptors, Adrenomedullin ; Receptors, Calcitonin ; genetics ; physiology ; Receptors, Calcitonin Gene-Related Peptide ; genetics ; physiology ; Receptors, Peptide ; genetics ; physiology ; Reverse Transcriptase Polymerase Chain Reaction