Genes, Reporter ; Genetic Vectors ; Luciferases ; genetics ; Transcription, Genetic ; bcl-2-Associated X Protein ; genetics

Objective To investigate the effect of edaravone on the JAK2/STAT3 signaling pathway after ischemia-reperfusion injury in donor rat liver under different cold ischemia times. Methods A total of 102 SD rats were randomly divided into sham operation group,control group and experimental group. Six rats were in sham operation group with free liver operation and no transplantation. Forty-eight rats were in control group and experimental group respectively, and divided into subgroups according to the different cold ischemia times (30 min, 6 h, 12 h and 18 h). There were 6 donors and 6 recipients in each group. The rat model of orthotopic liver transplantation was established by modifiedtwo cuff method. All the donors were perfused by abdominal aorta and the warm ischemia time was 3-5 min. After different cold ischemia times, the experimental group was treated with edaravone (3 mg/kg) at 5 min before the opening of the new hepatic artery, and control group was injected with 3 mg/kg saline. Recipients of each group were sacrificed after 6 h. Finally, real-time fluorescence quantitative PCR was used to analyze the relative expression of JAK2/STAT3 mRNA of donor liver. Results The GAPDH gene and JAK2/STAT3 were well amplified. Under the same cold ischemia time, compared with the control group, the relative expression of JAK2/STAT3 was significantly decreased in the experimental group (P<0.05). With the prolongation of cold ischemia time, the relative expressions of JAK2 and STAT3 mRNA showed a decreasing trend in control group and experimental group, while the relative expression of JAK2 mRNA increased first and then decreased in the experimental group (P<0.05). Conclusion Edaravone has a protective effect on transplanted donor liver during different cold ischemia times, and extends the cold ischemia time for 18 h, which may be related to the inhibition of JAK2/STAT3 signal transduction pathway.

Objective To explore the apoptosis of alveolar epithelial cell(AEC)and the dynamic changes of Caspase-3 mRNA and Bax and Bcl-2 expression in premature rat with hyperoxia-induced chronic lung disease(CLD).Methods Sixty premature rats within 1 day after birth were randomly divided into 2 groups:hyperoxia group(n=30)and control group(n=30).Hyperoxia-induced premature rat CLD models were prepared,and situ nick end-labeling(TUNEL),reverse transcription polymerasechain reaction(RT-PCR)and immunohistochemical technique were used to determine apoptosis index(AI)of AEC and the expressions of Caspase-3 mRNA and Bax and Bcl-2 proteins in lung tissues at 1,3,7,14 and 21 d after birth.Results Compared with control group,in the hyperoxia group,on the third day after exposured to hyperoxia,the AI of AEC and the expressions of Caspase-3 mRNA and Bax began to increase,and the expression of Caspase-3 mRNA was kept at high level on 7-21 d.The expression of Bcl-2 began to decrease on 7 d,and significantly decreased on 7-21 d.AI of AEC was positively correlated with the expression of Caspase-3 mRNA and Bax,and negatively with the expression of Bcl-2.Conclusions Hyperoxia may induce the increased expression of Caspase-3 mRNA,which might result in the abnormal expression of Bax and Bcl-2 in lung tissues and their imbalance,which might be one of the underlying mechanisms of apoptosis of AEC in premature rats with CLD.

Animals ; Coronary Disease ; Diagnosis, Differential ; Disease Models, Animal ; Medicine, Chinese Traditional ; Yang Deficiency

ObjectiveTo investigate the mechanism and the protective effect of heat shock protein 27 (HSP27) on rat cardiomyocytes when ischemic preconditioning performed.MethodsCultured rat cardiomyocytes were divided into four groups: control group, ischemic group,ischemic preconditioning group and cyclohexamide group. Cell viabilities were analyzed by MTT. The apoptosis was evaluated with DNA ladder and flow cytometry Annexin V Flous staining. Western Blot was used to determine the expression of HSP27 and caspase-3 in cardiomyocytes.ResultsIschemic preconditioning could improve cell viability. The apoptosis ratio in ischemic preconditioning group was significantly less than that in ischemic group. These were accompanied by an increase in the expression of HSP27 and a decrease in caspase-3. The expression of the increased HSP27 and the protective effect induced by ischemic preconditioning were completely abolished by the presence of cycloheximide, a translation inhibitor.ConclusionThe expression of HSP27 induced by ischemic preconditioning plays an important role in protecting cardiomyocytes, and the mechanism is possibly related to the inhibition of cell apoptosis.

Objective To analyze the differential proteomics of ASMC stimulated by wild IL-13 and mutant IL-13 and to investigate the relations of protein profiles of ASMC to asthma and possible targets for the treatment of bronchial asthma.Methods The total proteins of ASMC stimulated by wild IL-13 and mutant IL-13 were separated by immobilized pH gradient(IPG)-based 2-DE and the differentially expressed protein spots were identified by matrix assisted laser desorption-time of flight mass spectrometry(MALDI-TOF-MS). Results The 2-DE detected approximately(840?21)spots on wild IL-13 samples and(892?17)spots on mutant IL-13 samples(n=3)and(685?19)spots matched.Six significantly differential proteins were subjected to MALDI-TOF-MS analysis and three of them were identified as stathmin 1,Ribosomal protein p~0 and NADH dehydrogenase.Conclusions ASMCs stimulated by wild IL-13 and mutant IL-13 present different proteomic profiles that may shed some light on the mechanism for the asthma causing effect of wild IL-13 and mutant IL-13.

objective To study the reationship beween drug-resistance gene mutation and drug-resistance level in M. tuberculosis. Methods 108 M. tuberculosis clinical isolated strains from sputum specimens were analyzed by PCR-SSCP and traditional drug susceptibility tests. Results the gene mutation rate of SM, RFP, INH and EMBresistance climical isolated strains was 78.5%, 68.2%, 70.5% and 48.6% respectively, and the mutation rate of SM, RFP, INH and EMB high concentration resistance isolated strains was 86.5% , 89% , 84% and 48.6% respectively, but 28.5 % , 16.6% and 7.1% was the mutation rate of low concentration resistance strains. Conclusion The gene mutation was in relation with drug resistance level of M. tube rculosis. The gene mutation rate was hiher in high concentration resistance isolated strains than in low concentration resistane isolated strains.

Objective To analyze the clinical characteristic and prognosis of Serratia infections in newborn infants and increase awareness of Serratia infections.Methods The clinical manifestations,diagnosis,complications,treatment and prognosis of Serratia infections were analyzed in 4 hospitalized newborn infants in neonatology center from Jul.2008 to Feb.2009.Results Among the 4 cases,blood culture revealed Serratia marcescens in 3 cases(1 case was preterm infant),cerebral spinal fluid culture revealed Serratia liquefacien in the fourth case.The main clinical manifestations were fever,convulsion and poor response,WBC and CRP were much higher,while obvious thrombocytopenia was only found in the preterm infant.Two cases of septicemia infection alone recovered after the treatment of the third-generation cephalosporin for at least 2 weeks,while the other 2 cases of septicemia infection combined with purulent meningitis,included 1 case of preterm infant and 1 case of Serratia liquefacien infection,developed meningoencephalitis and brain abscess confirmed by serial imaging,both of which had poor neurological sequelaes.Conclusions As a opportunistic pathogen,Serratia can cause severe infection in newborns.The patients complicated with meningitis should be followed up and pay more attention to the high incidence of neurological sequelaes.

AIM: (1) To investigate the effect of amniotic membrane transplantation (AMT) on rabbit conjunctival surface reconstruction with severe alkali burns. (2) To evaluate the possibility of AMT treatment for ocular alkali burns during recovering stage.METHODS: Animal models were established on 30 eyes of rabbits by creating severe alkali burns on the conjunctiva from the upper corneal limbus to the upper conjunctival fornix.Preserved human amniotic membrane transplantations and reconstruction of conjunctival fornix were performed at one week after injury (recovering stage). Epithelium growth of burned area after transplantation was observed using light microscope at 1, 2, 3, 4, and 8 weeks. Conjunctival tissue in transplantation area was collected at 1, 4 and 8 weeks. The ultrastructure of the collected tissue was studied by electron microscope. The results were compared with control group,which received only vitamin C subconjunctival injection and antibiotic eye drops as treatment for alkali burn. Exterior eye pictures were also taken at the end of the observation, the width from upper corneal limbus to the edge of upper fornix was measured. Data was analyzed statistically.RESULTS: 1) Tn the transplant group, conjunctival epithelium growth was observed in the area of AMT under both light and electron microscope 1 week after surgery. At 4weeks, conjunctival epithelium with goblet cells that resembled normal conjunctival tissues was observed in the whole amniotic membrane area. At 12 weeks, the conjunctival epithelium on the amniotic membrane was well formed, and the connective tissue under the epithelium was loose at the fornix. No fibrosis was identified. In contrast, conjunctival epithelium necrosis was observed in the control group at 2weeks after alkali burns. Re-epithelization did not occur through the 12-week observation. Severe fibrosis with inflammatory cells infiltration was observed between 4 to 8weeks. At 12 weeks, fibrosis of the connective tissue at the fornix developed and there were no conjunctival epithelium covering the burned area. 2) In the transplant group, the conjunctiva in transplanted area had no scarring and appeared smooth at 12 weeks. Upper fornix was reconstructed. The depth of fornix was 7.9±0.3mm (7.6-8.2mm), which was approximate to the normal depth 8.2±0.2mm (8.0-8.4 mm,P＞.05). While in the control group, the burned area appeared rough with granuloma formation and severe scarring. Upper fornix became shallow. The depth of fornix was 3.1±1.7mm(1.0 to 4.5mm.), and significant difference was found between control and transplant group (P＜0.01).CONCLUSION: Human amniotic membrane preserved in glycerin can promote cell adhering, migrating and differentiating of normal conjunctival epithelium.Reconstruction of conjunctival surface in early stage of alkali burn can be achieved by AMT. AMT can effectively prevent symblepharon formation.

OBJECTIVETo investigate the effect of component I from agkistrodon acutus venomon (AAVC-I) the migration of human umbilical vein endothelial cells (HUVECs), and to elucidate the possible anti-angiogenic mechanism of AAVC-I.

METHODSThe effect of AAVC-I on the migration of HUVECs which was cultivated in vitro and treated with AAVC-1 at four concentrations: 0, 20, 40, 80 microg/ml, was observed by methods of scratch wound-healing and Transwell assay. The expression level of mRNA and protein of P-selectin and intercellular cell adhension molecule-I (ICAM-1) were examined by RT-PCR and Western blot assay.

RESULTSCompared with the blank group, the migration ability of HUVECs in each AAVE-I treated group was reduced in a dose-dependent manner, and the expression level of the mRNA and protein of P-selectin and ICAM-1 were decreased.

CONCLUSIONAAVC-I inhibits the migration of endothelial cell, which is acted by down-regulation of the expression content of mRNA and protein of P-selectin and ICAM-1.

Cell Movement ; drug effects ; Cells, Cultured ; Crotalid Venoms ; pharmacology ; Down-Regulation ; Human Umbilical Vein Endothelial Cells ; drug effects ; Humans ; Intercellular Adhesion Molecule-1 ; metabolism ; P-Selectin ; metabolism ; RNA, Messenger