Objective: IFN-y is produced by both activated T and NK cells in response to mitogen or antigen and has a broad range of immunoregulatory activity. IL-12 has been described as a strong inducer of IFN-?production and promotes the differentiation of naive CD4+T cells toward the Th1 phenotype, priming them for IFN-?production, and consequent induction of cell-mediated immunity. Aim is to know endogenous production of IL,12 from PBMC inducing production of IFN-?in vitro, which is involving in mechanism of T cells to be activated. Methods: Induced IFT-? secretion from human PBMC by stimulated with anti-CD3 , PHA, anti-CD3 plus anti-CD28 and antigen(MLC) Also inhibited IFN-?production by neutralizing antibodies to IL-12 and IL-12R?1 significantly. Results: IFN-?secretion from human activated PBMC is endogenous IL-12dependent, and activated T cells induce the production of IL-12 from APC by a mechanism involving the interaction between CD40L on T cells and CD40 on APC. Conclusion: These results suggest that endogenous IL-12 plays an important role in the normal host defense against infection by a variety of intracellular pathogens and also plays a central role in the genesis of some forms of immunopathology including autoimmune diseases and transplantation rejections.

Objective:To study the effect of IL-24 on the differentiation and function of osteoclasts.Methods:Mature osteoclasts were isolated from long bones of neonate rats.Optimal multiplicity of infection(MOI)of AdCMV-EGFP was determined.Then osteo-clasts and RAW264.7 cells were transfected with AdCMV-EGFP and AdCMV-IL-24 respectively.The function of osteoblasts was studied by the observation of bone resorption lacunae,the differentiation of RAW264.7 cells was evaluated by the expression of osteo-clast related genes examined with real-time PCR.Results:Osteoclasts were TRAP positive with more than 2 neclei.MOI=400 was suitable for AdCMV-EGFP transfection.The resorption lacunae area in AdCMV-IL24 transfected cells was larger than that in AdCMV-EGFP transfected cells(P<0.05).Real-time PCR showed that under induced conditions,osteoclastic related genes NFATc1,CTSK and TRAP were changed in RAW264.7 cells transfected with AdCMV-IL-24.Conclusion:IL-24 may promote the differentiation and function of osteoclasts.

Objective To investigate the influence of non-dipper blood pressure rhythm on peripheral atherosclerosis in elderly hypertensive patients.Methods The 199 elderly hypertensive patients with 24-hour average systolic blood pressure＜140 mmHg were selected.Body mass index (BMI),glycosylated hemoglobin,blood lipids,uric acid,creatinine,Brachial ankle pulse wave velocity (baPWV),ankle arm index(ABI) and 24-hour ambulatory blood pressure monitoring were tested and calculated.The elderly patients were divided into dipper hypertensive group (n=95),and non-dipper hypertensive group (n=104).The relationships of arteriosclerosis with lifestyle and the circadian rhythm of blood pressure (non-dipper hypertension) were analyzed.Results Circadian blood pressure rhythm,age,levels of total cholesterol,triglyceride and serum creatinine,nocturia times,and smoking index were significantly related with atherosclerosis.There was a relationship between circadian blood pressure rhythm and baPWV,with the greatest standardized regression coefficient of 0.439.There were obviously linear relationships between levels of serum creatinine,nocturia times,age and ABI.There were significant differences in baPWV,ABI and smoking index between dippers and non-dippers groups [(1746.0±246.9) vs.(2115.7±321.8),(1.14±0.10) vs.(1.11±0.18),(251.8±272.8) vs.(368.5±339.9),P＜0.05 or 0.001].The multiple linear regression analysis showed that smoking index was significantly correlated with baPWV,but not correlated with blood pressure rhythm.In the ROC curve of MAP difference percentage and baPWV diagnosis result,the area under the curve was 0.8188,the optimal cut-off was 10.9％,the sensitivity was 62.5％ and the specificity was 85.7％.In the ROC curve of MAP difference percentage and AMI diagnosis result,the area under the curve was 0.7589,the optimal cut-off was 8.1％,the sensitivity was 88％,and the specificity was 61％.These results reflected that there were better consistences between MAP difference percentage and baPWV,ABI,and indirectly indicted that there was a relationship between MAP difference percentage and atherosclerosis.Conclusions We advise the elderly patients to give up the bad habits,help them control the systolic blood pressure,and advise them to adjust the medication time according to the circadian rhythm of blood pressure of every elderly hypertensive patient in order to restore the normal circadian blood pressure rhythm and delay the atherosclerotic process.

Arsenites ; toxicity ; Cell Cycle ; drug effects ; Cell Division ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Dose-Response Relationship, Drug ; Humans ; Keratinocytes ; cytology ; Skin ; cytology ; Sodium Compounds ; toxicity

NMDA-induced excitotoxicity cause severe neuronal damage including apoptosis and necrosis. The present study was aimed to evaluate the proportion of NMDA-induced apoptosis of rat cortical neurons and discover signal transduction mechanism. Caspase inhibitor and lactate dehydrogenase (LDH) assay were used to study the NMDA-induced apoptosis. To explore the involved signal pathways, the primary culture of rat cortical neurons were pretreated by the inhibitors of three MAPK pathways, extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38 MAPK. With 2 h of NMDA treatment, cellular apoptosis was measured by caspase-3 activity, terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labeling (TUNEL) and Annexin V staining. The results showed that: (1) Caspase-dependent apoptosis accounted for 22.49% in NMDA-induced neuronal death; (2) Pretreatment with p38 MAPK inhibitor SB203580 (10 μmol/L) significantly decreased NMDA-mediated caspase-3 activity by 30.43% (P < 0.05). However, ERK inhibitor PD98059 (20 μmol/L) or JNK inhibitor SP600125 (20 μmol/L) did not influence caspase-3 activity; (3) Pretreatment with SB203580 significantly reduced the number of NMDA-induced TUNEL-positive cells by 33.10% (P < 0.05). PD98059 (20 μmol/L) or SP600125 (20 μmol/L) did not show obvious effect; (4) Pretreatment with SB203580 (10 μmol/L) significantly reduced the number of NMDA-induced early apoptotic neurons by 55.56% (P < 0.05). Also, SP600125 (20 μmol/L) significantly decreased the amount of late apoptotic/dead cells by 67.59% (P < 0.05). There was no effect of PD98059 (20 μmol/L). These results indicate that: (1) NMDA induces neuronal apoptosis besides necrosis; (2) p38 MAPK, but not JNK and ERK, is involved in NMDA-induced neuronal apoptosis, and inhibition of the apoptotic signaling pathway contributes to neuroprotection; (3) JNK activation might contribute to NMDA-induced neuronal necrosis rather than apoptosis.
Animals ; Anthracenes ; pharmacology ; Apoptosis ; Caspase 3 ; metabolism ; Cells, Cultured ; Extracellular Signal-Regulated MAP Kinases ; antagonists & inhibitors ; Imidazoles ; pharmacology ; JNK Mitogen-Activated Protein Kinases ; antagonists & inhibitors ; MAP Kinase Signaling System ; N-Methylaspartate ; pharmacology ; Neurons ; cytology ; Primary Cell Culture ; Pyridines ; pharmacology ; Rats ; p38 Mitogen-Activated Protein Kinases ; antagonists & inhibitors

OBJECTIVE: To investigate the involvement of ephrinB2 in periodontal tissue remodeling in compression areas during orthodontic tooth movement and the effects of compressive force on EphB4 and ephrinB2 expression in osteoblasts and osteoclasts. METHODS: A rat model of experimental tooth movement was established to examine the histological changes and the localization of ephrinB2 in compressed periodontal tissues during experimental tooth movement. RAW264.7 cells and ST2 cells, used as precursor cells of osteoclasts and osteoblasts, respectively, were subjected to compressive force in vitro. The gene expression of EphB4 and ephrinB2, as well as bone-associated factors including Runx2, Sp7, NFATc1, and calcitonin receptor, were examined by quantitative real-time polymerase chain reaction (PCR). RESULTS: Histological examination of the compression areas of alveolar bone from experimental rats showed that osteoclastogenic activities were promoted while osteogenic activities were inhibited. Immunohistochemistry revealed that ephrinB2 was strongly expressed in osteoclasts in these areas. Quantitative real-time PCR showed that mRNA levels of NFATc1, calcitonin receptor, and ephrinB2 were increased significantly in compressed RAW264.7 cells, and the expression of ephrinB2, EphB4, Sp7, and Runx2 was decreased significantly in compressed ST2 cells. CONCLUSIONS: Our results indicate that compressive force can regulate EphB4 and ephrinB2 expression in osteoblasts and osteoclasts, which might contribute to alveolar bone resorption in compression areas during orthodontic tooth movement.
Animals ; Bone Resorption* ; Gene Expression ; Immunohistochemistry ; Models, Animal ; Osteoblasts* ; Osteoclasts* ; Rats ; Real-Time Polymerase Chain Reaction ; Receptors, Calcitonin ; RNA, Messenger ; Tooth Movement*

The aim of this study is to identify the present status of the scientific and technological personnel in the field of traditional Chinese medicine (TCM) resource science. Based on the data from Chinese scientific research paper, an investigation regarding the number of the personnel, the distribution, their output of paper, their scientific research teams, high-yield authors and high-cited authors was conducted. The study covers seven subfields of traditional Chinese medicine identification, quality standard, Chinese medicine cultivation, harvest processing of TCM, market development and resource protection and resource management, as well as 82 widely used Chinese medicine species, such as Ginseng and Radix Astragali. One hundred and fifteen domain authority experts were selected based on the data of high-yield authors and high-cited authors. The database system platform "Skilled Scientific and Technological Personnel in the field of Traditional Chinese Medicine Resource Science-Chinese papers" was established. This platform successfully provided the retrieval result of the personnel, output of paper, and their core research team by input the study field, year, and Chinese medicine species. The investigation provides basic data of scientific and technological personnel in the field of traditional Chinese medicine resource science for administrative agencies and also evidence for the selection of scientific and technological personnel and construction of scientific research teams.
Bibliography of Medicine ; Biomedical Research ; manpower ; Databases, Factual ; Humans ; Laboratory Personnel ; Medicine, Chinese Traditional ; methods ; Plants, Medicinal ; chemistry ; growth & development

OBJECTIVETo study mechanism of the apoptosis of rat pancreas islet β cell strain (INS-1 cells) induced by sodium arsenite.

METHODSINS-1 cells were exposed to sodium arsenite at the different concentrations. MTT assay was used to detect the viability of INS-1 cells. The potentials on mitochondrial membrane and lysosome membrane of INS-1 cells were determined with the fluorescence spectrophotometer. The apoptotic levels of INS-1 cells exposed to sodium arsenite were observed by a fluorescence microscope and flow cytometry.

RESULTSAfter exposure to sodium arsenite, the viability of INS-1 cells significantly decreased with the doses of sodium arsenite. At 24 h after exposure, the OD values of the mitochondrial membrane potentials declined observably with the doses of sodium arsenite (P < 0.01). At 48 h after exposure, the OD values of the lysosome membrane potentials significantly increased with the doses of sodium arsenite (P < 0.01). At 72 h after exposure, the apoptotic cells were observed under a fluorescence microscope and enhanced with the doses of sodium arsenite. The apoptosis cells with light blue, karyopyknosis, karyorrhexis, apoptotic body and chromatin concentration appeared. The results detected with flow cytometry indicated that after exposure, the apoptotic INS-1E cells significantly increased with the doses of sodium arsenite.

CONCLUSIONSThe sodium arsenite can induce the apoptosis of INS-1 cells through the mitochondria-lysosome pathway.

Animals ; Apoptosis ; drug effects ; Arsenites ; toxicity ; Cells, Cultured ; Insulin-Secreting Cells ; drug effects ; Lysosomes ; metabolism ; Membrane Potentials ; drug effects ; Mitochondria ; metabolism ; Rats ; Sodium Compounds ; toxicity

OBJECTIVETo construct mouse enhanced green fluorecence protein (EGFP) -peroxisome proliferator-activated receptor (PPAR)gamma2, and to detect EGFP-PPARgamma2 expression in infected mouse bone marrow mesenchymal stem cells (BMSC).

METHODSCut the fragment of PPARgamma2 from the expression plasmid pcDNA flag PPARgamma2, then cloned the gene fragment into pEGFP-C1 and pEGFP-N1 vector. Subsequently, subclone the fragment EGFP-PPARgamma2 from pEGFP-C1-PPARgamma2 into the shuttle plasmid DC315. HEK293 cells were co-transfected with the constructed recombinant shuttle plasmid DC315-EGFP-PPARgamma2 and large adenovirus helper plasmid pBHGlox deltaE1, 3Cre in mediation of liposome. The obtained replication-defective recombinant adenovirus Ad-EGFP-PPARgamma2 was confirmed. Then it was propagated in HEK293 cells. After the BMSC were transfected for 72 h, adipogenic differentiation was demonstrated.

RESULTSHEK293 cells were transfected with the pEGFP-C1-PPARgamma2 or pEGFP-N1-PPARgamma2 in mediation of liposome. The former green fluorescence protein was better than the latter by fluorescence microscope. The recombinant plasmids were digested and identified. Western blot analysis showed the expression of EGFP-PPARgamma2 in vitro. EGFP-PPARgamma2 protein was detectable in the nucleus of BMSC.

CONCLUSIONThe recombinant adenovirus encoding EGFP-PPARgamma2 fusion protein was successfully constructed, which provided a basis for application of EGFP-PPARgamma2 gene to adenovirus-mediated gene therapy.

Adenoviridae ; Animals ; Bone Marrow Cells ; metabolism ; Genetic Vectors ; Green Fluorescent Proteins ; metabolism ; HEK293 Cells ; Humans ; Mesenchymal Stromal Cells ; metabolism ; Mice ; PPAR gamma ; metabolism ; Recombinant Proteins ; metabolism ; Transfection

Objective:To investigate the role of metformin (MET) in inflammatory response in abdominal aortic aneurysm(AAA) formation induced by angiotensin Ⅱ.Methods:AAA models were established by Ang Ⅱ infusion in ApoE-/-mice.Thirty male ApoE-/-mice (8-10-week-old) were randomly and equally divided into four groups:Control group,AAA group and AAA+MET group.HE staining and immunohistochemical staining were used to analyze the inflammatory response.Real-time quantitative polymerase chain reaction (qPCR) was used to analyze the mRNA expression of intercellular cell adhesion molecule-1 (ICAM-1).Western blotting was used to analyze the activation of nuclear factor κB signaling (NF-κB).Results:The AAA incidence and mean maximum suprarenal aortic diameter of AAA group is larger than the AAA+MET group.Compared with the AAA grouP,the expression of ICAM-1 and activity of NF-κB is lower in AAA+MET group.Conclusion:Metformin can attenuate AAA formation partially by regulating macrophage infiltration through regulating the mRNA expression of ICAM-1 regulated by the activation of NF-κB signaling pathway.