Objective: To observe the protective effect of ischemic post-conditioning on myocardial reperfusion injury with the potential mechanism in experimental rabbits. Methods: A total of 36 healthy New Zealand rabbits were divided into 6 groups:①Sham group,②Ischemic reperfusion control (CON) group,③Myocardial ischemic post-conditioning (MpostC) group,④Remote ischemic post-conditioning (RPostC) group,⑤MPostC+5-HD group,⑥RPostC+5-HD group.n=6 in each group. The ischemic reperfusion injury model was established by left ventricular descending artery occlusion for 45 min followed by reperfusion for 120 min. Bilateral external iliac artery was occluded for 5 min to induce the short skeletal muscle ischemia. The indexes of cardiac function and plasma CK , LDH activities were measured at baseline, end of ischemia and 1, 2 h after reperfusion respectively, the sizes of myocardial infarction (MI) were examined and compared among different groups. Results: ①Compared with CON group, the indexes of cardiac function were improved in MPostC and RPostC groups at 1, 2 h after reperfusion,P<0.05, and compared with MPostC group and RPostC group, the indexes of cardiac function were improved in MPostC+5-HD and RPostC+5-HD groups,P<0.05.②Compared with CON group, plasma CK, LDH activities were decreased in MPostC and RPostC groups at 2 h after reperfusion,P<0.05, and compared with MPostC group and RPostC group, the CK, LDH activities were increased in MPostC+5-HD and RPostC+5-HD groups,P<0.05.③The ischemic ranges and areas were similar between CON group and the other 5 groupsP>0.05. The MI ranges and areas in MPostC and RPostC groups were much less than that in CON group,P<0.05, and compared with MPostC group and RPostC group, the MI ranges and areas increased in MPostC+5-HD and RPostC+5-HD groups,P<0.05. The MI ranges and areas were similar between CON group and MPostC+5-HD, RPostC+5-HD groups,P>0.05. Conclusion: Classical ischemic post-conditioning and remote organ ischemic post-conditioning both have protective effect on myocardial reperfusion injury in experimental rabbit, which might be related to the activation of mitochondrial ATP-sensitive potassium channels.

Objective: To investigate the preventive effect of alprostadil (prostaglandin E1) combining hydration therapy on contrast induced nephropathy (CIN) after percutaneous coronary intervention (PCI) in elder patients. Methods: A total of 175 patients with coronary artery disease (CAD) elder than 75 years received PCI in our hospital from 2011-03 to 2014-03 were retrospectively studied, those including 122/175 (69.71 %) with male gender. The patients were at the mean age of (74.7 ± 3.9) years and randomly allocated into 2 groups: Control group, based on routine treatment, the patients received regular hydration of intravenous normal saline 1 ml/(kg?h) at 6 hours prior and 12 hours after PCI,n=84 and Alprostadil+hydration group, based on routine treatment and regular hydration, the patients received intravenous alprostadil 10μg in 100ml normal saline twice a day at 1 day prior PCI and the 3rd day after PCI,n=91. The serum levels of creatinine (SCr) were examined prior PCI and 3 days continuously after PCI, creatinine clearance rates (Ccr) were calculated and the CIN occurrence rates were compared between 2 groups. Results: At the 3rd day after PCI, the mean SCr level in Alprostadil+hydration group (92.08 ± 21.65) μmol/L was lower than Control group (99.43 ± 22.77) μmol/L,P<0.05, the mean Ccr level in Alprostadil+hydration group (63.78 ± 20.58) ml/min was higher than Control group (57.09 ± 22.31) ml/min,P<0.05. The occurrence rate of CIN after PCI was higher in Control group (13.1%, 11/84 patients) than Alprostadil+hydration group (3.3%, 3/91 patients),P<0.05. Conclusion: Alprostadil combining hydration therapy may obviously protect renal function and reduce the incidence rate of CIN in elder CAD patients after PCI treatment.

AIM: To prepare and characterize a mouse anti-human CD40 mutant monoclonal mAb. METHODS: Female BALB/c mice of 6-8 weeks old were immunized with CD40 mutant transfectant (L929-CD40mu) as immunogen. The spleen B cells of the mice were fused with Sp2/0 myeloma cells. The hybridoma cells were screened with CD40 mutant transfectant (L929-CD40mu) by FCM. Faststrip analysis was performed to identify Ig subclass of this mAb. The epitope recognized by this mAb was detected by Bio-5C11 competitive assay. Western blot technique was adopted to identify the mAb. The proliferation of tumor cells in vitro was analyzed by MTT assay and apoptosis of tumor cells in vitro was analyzed by PI-annexin V assay. RESULTS: One hybridoma cell line named 10C5 was obtained, which had the property of secreting anti-human CD40 mutant monoclonal antibody continuously and steadily. This mAb specifically recognized human CD40 mutant molecule and induced the apoptosis of tumor cells in vitro. CONCLUSION: One hybridoma cell line which can secret a mouse anti-human CD40 mutant mAb has been prepared successfully. This mAb can inhibit the growth of tumor cells expressing CD40 mutant and induce their apoptosis in vitro.

Objective: To study the effect of miR-194-3p on the migration of keloid fibroblasts. Methods: Differentially expressed miRNA were screened by gene chip in 8 human keloid and normal tissues. The down regulated miR-194-3p was selected for study and its binding to RUNX2 was predicted by MiRDB, and verified by fluorescent reporter gene in human keloid fibroblasts (HKFs) and passage 3 keloid cells, respectively. The effect of miR-194-3p on the migration of fibroblasts was detected by transwell assay. Western blot and real-time PCR were used to analyze the effect of miR-194-3p on RUNX2 and MMP2 expression in HKFs. The results were analyzed by SPSS 19.0 software and compared by non-paired t test. P<0.05 was considered statistically significant. Results: There were abnormal expressions of miR-721, miR-21-3p, miR-382-3p, miR-194-3p, miR-3107-5p and miR-144-5p in keloid. miRDB software analysis showed that miR-194-3p and RUNX2 had targeted binding sites. Reporter gene experiments showed that miR-194-3p inhibited the activity of RUNX2 by about 50% compared with the control group (t=2.7764, P=0.0005). MiR-194-3p could significantly inhibit the expression of RUNX2 and MMP2, and inhibited the migration of keloid fibroblasts (t=2.7764, P<0.05). Conclusion MiR-194-3p may inhibit the migration of fibroblasts by inhibiting the activity of RUNX2 in keloid.

BACKGROUND:Inflammation and oxidative stress contribute to the barriers of regeneration in chronic wound.Oxymatrine has various biological activities,such as anti-oxidation,anti-inflammation and so on,which may have the potential effect of promoting wound healing. OBJECTIVE:To investigate the effect of oxymatrine on wound healing and the protective effect on H2O2-induced oxidative stress injury in human keratinoid cell line HaCaT cells. METHODS:(1)In vivo experiment:Hyaluronic acid methacryloyl hydrogels containing 0,0.05,0.1,0.2 g/L oxymatrine were prepared.A full-layer skin defect model with a diameter of 12 mm was made in the back of 75 diabetic mice and randomly divided into five groups for intervention,with 15 mice in each group.The wounds of the model group were bandaged and fixed.The wounds of the hydrogel group were covered with hyaluronic acid methacryloyl hydrogel.The wounds of the low-dose,moderate-dose and high-dose oxymatrine groups were covered with hyaluronic acid methacryloyl hydrogel containing 0.05,0.1,and 0.2 g/L oxymatrine,respectively,and then bandaged and fixed after light curing.Relevant indicators were detected within 14 days.(2)In vitro experiment:Human keratinocyte line HaCaT was divided into five groups.The normal group was cultured conventionally.H2O2 group and low-,moderate-and high-concentration oxymatrine groups were treated with H2O2 for 4 hours,and then the medium was replaced with medium containing 0,0.05,0.1,and 0.2 g/L oxymatrine,respectively,and the relevant indexes were detected after 24 hours of culture. RESULTS AND CONCLUSION:(1)In vivo experiment:Compared with the model group,the wound healing rate of mice in the hydrogel group had no significant change.The wound healing rate of mice in the low-,moderate-and high-dose oxymatrine group was increased at 7 and 14 days after treatment(P<0.05).Pathological observation of wound section 14 days after treatment showed that compared with the model group,the thickness of regenerated epidermal layer,the number of microvessels,and collagen deposition in the moderate-and high-dose oxymatrine groups were increased(P<0.05).Western blot assay analysis of wound samples 7 days after surgery showed that compared with the model group,the protein expressions of tumor necrosis factor α and interleukin 6 in the moderate-and high-dose oxymatrine groups were decreased(P<0.05).(2)In vitro experiment:CCK8 assay,EdU and Ki67 staining showed that compared with the H2O2 group,the cell proliferation ability of the moderate-and high-concentration oxymatrine groups was significantly increased(P<0.05).Compared with the H2O2 group,mitochondrial membrane potential was increased(P<0.05)and reactive oxygen species content was decreased(P<0.05)in the moderate-and high-concentration oxymatrine groups.Western blot assay results showed that compared with the H2O2 group,the expression levels of Nrf2 nuclear protein,Nrf2 total protein,HO-1 protein,and superoxide dismutase 1 protein were increased in the high-concentration oxymatrine group(P<0.05).(3)These findings confirm that oxymatrine can alleviate oxidative stress damage in HaCat cells and accelerate wound healing by upregulating the levels of Nrf2 and HO-1 protein.