Objective:To study the mechanism of immune tolerance induce by protal venous injection of donor spleen cells on the dog model of renal transplantation.Methods:The donor spleen cells were injected through the protal vein during operation,one week later,renal transplantation was performed.IL-2 and IL-6 were studied by method of ELISA.Results:Level of IL-2 and IL-6 in protal venous group and cyclosporin group was higher than that of control group.There were no difference between protal venous group and cyclosporin group.Conclusion:Immune tolerance could be produced by protal venous injection of donor spleen cells.

Due to their small molecular weight,strong penetrating power,wide target range,strong ability of binding targets,stable quality,little immunogenicity,easiness to be synthesized and modified,and functional roles in molecular recognition and signal transduction,nucleic acid aptamers are now used as tools for molecular recognition and drugs delivery for the diagnosis and treatment of many diseases.

In the paper, iridology and computer-aided iridiagnosis technologies are briefly introduced and the extraction method of the collarette contour is then investigated. The iris map can be overlapped on the original iris image based on collarette contour extraction. The research on collarette contour extraction and iris map overlap is of great importance to computer-aided iridiagnosis technologies.
Diagnosis, Computer-Assisted ; methods ; Humans ; Image Processing, Computer-Assisted ; methods ; Iris Diseases ; diagnosis ; Software

Objective To explore the effect of transforming growth factor β1 (TGF-β1) on epigenetic histone lysine acetylation in the plasminogen activator inhibitor 1 (PAI-1) promoter and transcribe regions in glomerular mesangial cells (GMCs).Methods Chromatin immunoprecipitation assay and real-time quantitative PCR were used to detect Histone3K9 acetylation (H3K9Ac) in the PAI-1 promoter and transcribe regions induced by TGF-β1 and high glucose.Immunoprecipitation was also used to see the cooperation of Smad3,CBP and Sp1 proteins.Results In the four target regions of PAI-1 promoter,TGF-β1 treatment enhanced H3K9Ac at P1,P2 and P3 in GMCs (P ＜ 0.05),but no change was seen in the P4 region which was far from the transcription starting site.TGF-β1 obviously induced H3K9Ac in the T1 transcribe region of PAI-1 instead of T2 (P ＜ 0.05).High glucose increased PAI-1 mRNA expression and H3K9Ac around P1 promoter region (P＜ 0.05).TGF-β1 neutralizing antibody abrogated high glucose-induced H3K9Ac at PAI-1 promoter (P ＜ 0.01).TGF-β1 treatment could recruit Smad3 and CBP protein binding to the PAI-1 promoter regions (P1,P2,P3),and induce their cooperation in GMCs,which were responsing to TGF-β1 associated H3K9Ac.Conclusion TGF-β1 can induce H3K9Ac in the promoter and transcribe regions of PAI-1,promote Smad3 recruition and cooperation with Sp1 and CBP,which are associated with PAI-1 gene's regulation in GMCs.

This study is to evaluate the therapeutic effect of fibroblast growth factor 21 (FGF21) on NAFLD in MSG-IR mice and to provide mechanism insights into its therapeutic effect. The MSG-IR mice with insulin resistance were treated with high dose (0.1 micromol.kg-1d-1) and low dose (0.025 micromol.kg-1d-1) of FGF21 once a day for 5 weeks. Body weight was measured weekly. At the end of the experiment, serum lipids, insulin and aminotransferases were measured. Hepatic steatosis was observed. The expression of key genes regulating energy metabolism were detected by real-time PCR. The results showed that after 5 weeks treatment, both doses of FGF21 reduced body weight (P<0.01), corrected dyslipidemia (P<0.01), reversed steatosis and restored the liver morphology in the MSG model mice and significantly ameliorated insulin resistance. Additionally, real-time PCR showed that FGF21 significantly reduced transcription levels of fat synthetic genes, decreased fat synthesis and promoted lipolysis and energy metabolism by up-regulating key genes of lipolysis, thereby liver fat accumulation was reduced and liver function was restored to normal levels. In conclusion, FGF21 significantly reduces body weight of the MSG-IR mice, ameliorates insulin resistance, reverses hepatic steatosis. These findings provide a theoretical support for clinical application of FGF21 as a novel therapeutics for treatment of NAFLD.
Animals ; Body Weight ; drug effects ; Dose-Response Relationship, Drug ; Dyslipidemias ; metabolism ; Energy Metabolism ; drug effects ; Fatty Liver ; chemically induced ; complications ; Female ; Fibroblast Growth Factors ; administration & dosage ; pharmacology ; therapeutic use ; Insulin Resistance ; Lipolysis ; drug effects ; Liver ; metabolism ; pathology ; Male ; Mice ; Non-alcoholic Fatty Liver Disease ; drug therapy ; Sodium Glutamate

Adrenal Cortex Neoplasms ; complications ; Adrenal Gland Neoplasms ; complications ; Adrenal Glands ; pathology ; Adrenocortical Adenoma ; complications ; Humans ; Pheochromocytoma ; complications

Epiberberine, one of the most important isoquinoline alkaloid in Coptidis Rhizoma, possesses extensive pharmacological activities. In this paper, the liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used to study phase I and phase II metabolites. A Thermo HPLC system (including Surveyor AS, Surveyor LC Pump, Surveyor PDA. USA) was used. The cocktail probe drugs method was imposed to determine the content change of metoprolol, dapsone, phenacetin, chlorzoxazone and tolbutamide simultaneously for evaluating the activity of CYP2D6, CYP3A4, CYP1A2, CYP2E1 and CYP2C9 under different concentrations of epiberberine in rat liver microsomes. The result showed that epiberberine may have phase I and phase II metabolism in the rat liver and two metabolites in phase I and three metabolites in phase II are identified in the temperature incubation system of in vitro liver microsomes. Epiberberine showed significant inhibition on CYP2D6 with IC50 value of 35.22 μmol L(-1), but had no obvious inhibiting effect on the activities of CYP3A4, CYP1A2, CYP2E1 and CYP2C9. The results indicated that epiberberine may be caused drug interactions based on CYP2D6 enzyme. This study aims to provide a reliable experimental basis for its further research and development of epiberberine.
Animals ; Berberine ; analogs & derivatives ; chemistry ; metabolism ; Chromatography, High Pressure Liquid ; Cytochrome P-450 CYP2D6 ; metabolism ; Cytochrome P-450 CYP2D6 Inhibitors ; chemistry ; metabolism ; Drugs, Chinese Herbal ; chemistry ; metabolism ; Male ; Microsomes, Liver ; drug effects ; enzymology ; metabolism ; Molecular Structure ; Rats ; Rats, Sprague-Dawley ; Tandem Mass Spectrometry

Objective:Iron accumulation is related to the occurrence of postmenopausal osteoporosis. Meanwhile, autophagy abnormality of bone marrow hematopoietic cells is observed in hip osteoporotic fracture. This study is performed to investigate correlation between iron accumulation induced bone loss and hematopoietic autophagy dysfunction to explore the new risk factor of osteoporosis.Methods:Male iron accumulation mice model was established by intraperitoneally injecting ferric ammonium citrate. Serum ferritin and osteogenic indicator P1NP were tested by ELISA. Bone mineral density was measured by micro-CT. Femur and tibia bone marrows were collected for hematopoietic stem and progenitor cells proportion and cell apoptosis analysis. Autolysosome formation was measured by image flow cytometry. We used conditional mouse model Atg7 flox/flox; Vav-Cre(Atg7 -/-) in which Atg7 had been genetically deleted in the hematopoietic system. Bone marrow hematopoietic stem and progenitor cells were collected for RNA sequence. micro-CT scan was conducted for Atg7 -/- femur. Results:Ferritin level of iron accumulation mice was significantly higher than control group( P<0.05). Iron accumulation inhibited P1NP and induced decreased bone mineral density( P<0.05). Iron accumulation bone marrow displayed enhanced hematopoietic stem and progenitor cells proportion( P<0.05), with more cell apoptosis( P<0.05). Hematopoietic autophagy was deteriorated in iron accumulation bone marrow. Transcriptomic profiling showed up-regulation of iron activity in Atg7 -/- mice, with increased iron homeostasis and iron membrane transporter genes, including Lcn2, Tfr2, Slc40a1(Fpn1), Steap3, and Cpox. micro-CT revealed severe bone loss and decreased bone mineral density in Atg7 -/- mice( P<0.05). Conclusion:Iron accumulation induced bone loss is related to inhibition of hematopoietic cells. Hematopoietic autophagy dysfunction is associated with bone loss.

OBJECTIVETo explore the effect of cardiopulmonary bypass (CPB) on the profile of protein expression in peripheral blood mononuclear cells (PBMCs).

METHODSEleven patients undergoing cardiac surgery under cardiopulmonary bypass were enrolled in the study. Peripheral blood samples were collected before CPB (T0), 1 h after CPB (T1) and at the end of operation (T2), and PBMCs were obtained by gradient centrifugation. The profile of protein expression was analyzed using 2-D gel electrophoresis (2-DE) and mass spectrometry. The candidate proteins were further identified by Western blotting.

RESULTCompared to protein profile at T0, 12 protein spots were identified to be up-regulated in PBMCs at T1 (P <0.05), among which S100A9 reached the peak level at T1 and decreased after operation,but not returned to its initial level.

CONCLUSIONResults indicate that 12 proteins are likely to be involved in CPB, however, their roles need to be elucidated.

Adult ; Cardiopulmonary Bypass ; Electrophoresis, Gel, Two-Dimensional ; Female ; Humans ; Leukocytes, Mononuclear ; metabolism ; Male ; Mass Spectrometry ; Middle Aged ; Proteome ; metabolism ; Proteomics ; Young Adult

BACKGROUNDCathepsin B plays an important role in cell cycle, extracellular matrix changes and cutaneous tumorigenesis: whether it plays a role in photoaged skin remains unknown. This study aimed to investigate the role of cathepsin B in skin photoaging in vivo and in vitro.

METHODSThe expressions of cathepsin B were compared with immunohistochemical methods in solar exposed skin and solar protected skin of six healthy Chinese volunteers. The mRNA and protein expression of cathepsin B in ultraviolet light A (UVA) induced premature senescence fibroblasts in vitro were detected by real-time reverse transcription polymerase chain reaction (RT-PCR) and Western blotting technique.

RESULTSDecreased expression of cathepsin B was observed in photoaged skin compared with that of the solar protected skin. In the UVA induced, premature senescence fibroblasts, a lower expression of cathepsin B was detected by Western blotting and a decreased synthesis of cathepsin B mRNA in the same cells was revealed by real-time RT-PCR.

CONCLUSIONSThe results demonstrated a significant negative correlation between skin photoaging and cathepsin B in vitro and in vivo. We propose that cathepsin B, besides matrix metalloproteinases and antioxidant enzymes, is involved in the process of skin photoaging in that it contributes to extracellular matrix remodelling and is a dominant protease in cellular apoptosis and senescence.

Blotting, Western ; Cathepsin B ; analysis ; genetics ; physiology ; Female ; Fibroblasts ; radiation effects ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Skin ; radiation effects ; Skin Aging ; Ultraviolet Rays ; beta-Galactosidase ; analysis