Objective To investigate the effect of different doses of dexmedetomidine (DEX) on myocardial ischemia-reperfusion injury (MIRI) in rats.Methods Seventy-five SD male rats weighing between 250 and 300g were divided into sham group,MIRI group,small-dose DEX group,medium-dose DEX group and high-dose group according to the random number table,with 15 rats per group.Threading the left anterior descending coronary artery was done only in sham group,but the MIRI model was produced in the rest groups by ligation of the artery for 30 minutes followed by 120 minutes of reperfusion.Fifteen minutes before the ligation,small-,medium-and high-dose DEX groups were injected 2.5,5 and 10 μg · kg-1 · h-1 of DEX respectively until the end of reperfusion.Instead,an equal volume of normal saline was given in sham and MIRI groups.At the end of reperfusion,five rats in each group were used to determine the myocardial infarct size,and arterial blood samples and myocardial tissues from ten rats in each group were used to measure serum levels of interleukin-1β (IL-β) and serum tumor necrosis factor-α (TNF-α),expression of myocardial nuclear factor kappa B p65 (NF-κB p65) and change of myocardial pathomorphology.Results Myocardial infract size,degree of myocardial pathomorphology structure damage,serum levels of IL-1 β and TNF-αt and expression of myocardial NF-κB p65 in sham group were significantly lower in sham group than other groups (P ＜ 0.05).Above mentioned parameters in small-,medium-and high-dose DEX groups were all significantly decreased compared to MIRI group (P ＜ 0.05),and the decrease was most significant in medium-dose DEX group (P ＜ 0.05).Conclusions DEX can attenuate the MIRI in rats and the possible mechanism is suppressing the release of NF-κB p65,which can reduce serum pro-inflammatory cytokines like TNF-α and IL-1β.And mediumdose DEX exhibits better protective effect.

Objective To study the mechanism of killing and apoptosis-inducing effects of Capparis spinosa alkaloid (CSA) on human hepatocarcinoma cell line HepG2. Methods The killing effect of CSA on human hepatocarcinoma cell line HepG2 was studied by MTT method. Morphological observation of HepG2 cells was carried out by fluorescence microscope. Results The CSA had obvious cytotoxicity on the HepG2 in a dose-dependent manner and its IC50 value was 142.82 ?g/mL. The HepG2 cells showed the characteristic morphologic changes of apoptosis by the function of CSA and the apoptosis percentage is higher than that of the natural one. The progress of cells cycle from S phase to G2 phase had been blocked by CSA. The intracellular Ca2+ level had been increased by the function of CSA, which was positively related with drug concentration. Conclusion CSA has obviously killing and apoptosis-inducing effects on human hepatocarcinoma cell line HepG2 and calcium overload might also be invovled in these events.

Objective To explore the effects of transplantated bone marrow mesenchymal stem cells (MSCs) on myocardial fibrosis with the aid of ultrasound-mediated microbubbles (MB) and bispecific antibody(BiAb) combination.Methods With the aid of MB,isolated MSCs from male mice and the BiAb were transfused into female mice with isoproterenol-induced myocardial fibrosis via tail vein (MSCs + BiAb + MB group).BiAb was producted with anti-CD29 which can recognize MSCs and anti-myosin light chain antibody (AMLCA) which can specifically bind to injured myocardium.There were six groups investigated:MSC + BiAb + MB,MSC,BiAb,MB,MSC + BiAb,untreated,and control groups.Five weeks after treatment administration,the expressions of sex-determining region of Y-chromosome (SRY) in myocardium were detected by fluorescent quantitative PCR.The distribution of collagen was observed by sirius red staining.Heat shock protein-70 (HSP-70) in myocardium was detected by immunohistochemistry.Results The highest homing number of MSCs was in the MSCs + BiAb + MB group,MSCs + BiAb group ranked the second,and lowest in MSCs group.Compared to the untreated group,the MSCs + BiAb + MB,MSCs + BiAb and MSCs groups had less collagen deposition (P ＜0.05),and decreased level of HSP-70.Compared to those of the MSCs group,the collagen deposition were decreased in MSCs + BiAb + MB group (P ＜0.05).Conclusions MB and BiAb can promote the homing number of MSCs in mice.MB can further the homing rate and the repairing efficacy of MSCs.The homing MSCs can prevent the process of myocardial fibrosis.And HSP-70 was involved in the internal mechanism.

Conjugal plasmid pGH112 has been developed based on the replicons of Streptomyces coelicolor plasmid SCP2 and E. coli ColE. The plasmid contains ampicilin resistance gene(amp) for selection in E. coli and thiostrepton resistance gene (tsr) for selection in Streptomycetes, and a 0.76 kb oriT fragment of (IncP) RK2. Conjugal transfer of pGH112 was performed from E. coli to S. coelicolor A3(2), S. avermitilis, S. lividans TK54, S. toxytricini NNRL15443, S. venezuelae ISP5230 and Sacc. erythraea by conjugation, results show that the plasmid was able to transfer efficenctly from E. coli to Streptomycetes, was stably inherited in the recipients. pGH113 was constructed from pGH112 by combining the constitutive ermE promoter with green fluorescent protein gene(gfp).
Ampicillin Resistance ; genetics ; Conjugation, Genetic ; Escherichia coli ; genetics ; Green Fluorescent Proteins ; genetics ; Plasmids ; Streptomycetaceae ; genetics ; Thiostrepton ; pharmacology

OBJECTIVETo investigate the early diagnosis and treatment of acute mesenteric ischemia.

METHODSForty-two patients with acute mesenteric ischemia from June 2007 to November 2011 were reviewed retrospectively. All patients were diagnosed with DSA and (or) CT and (or) surgery. In this group, there were 32 cases of acute occlusion of meseteric ischemia (AOMI), 9 cases of superior mesenteric venous thrombosis (SMVT) and 1 case of non-occlusive mesenteric ischemia. The patients were treated using comprehensive treatment including early intervention treatment and application of the principle of damage control. The survival of all patients was followed up for 6 months or more in outpatient.

RESULTS(1) Of the 32 AOMI cases, 4 cases healing by systemic anticoagulation; The 19 cases received interventional treatment, including 10 cases received simply interventional treatment, surgery after the failure of intervention in 5 cases, 3 patients died without surgery and postoperative interventional treatment one cases were cured; Eight cases received surgery treatment; One case gave up. (2) Of the 9 SMVT cases, 2 cases healing by systemic anticoagulation; The 6 cases received interventional treatment, including 1 cases received simply interventional treatment, surgery after the failure of intervention in 1 cases, 4 cases to consider intestinal necrosis received interventional treatment again after surgery; One patient died without treatment. (3) Eight cases received delay abdomen close treatment with the principle of damage control surgery. The overall mortality rate of 23.8% (10/42). Interventional treatment of 26 cases, 4 deaths, a mortality rate of 15.3%; The abdomen delayed close of 8 cases, 1 death.

CONCLUSIONSThe results show that early diagnosis and treatment is critical to reduce AMI mortality. Comprehensive treatment of early intervention treatment and application of the principle of damage control can significantly reduce the mortality of AMI.

Adult ; Aged ; Aged, 80 and over ; Early Diagnosis ; Female ; Humans ; Ischemia ; diagnosis ; therapy ; Male ; Mesenteric Ischemia ; Middle Aged ; Retrospective Studies ; Tomography, X-Ray Computed ; Vascular Diseases ; diagnosis ; therapy ; Young Adult

OBJECTIVETo establish a routine procedure for the detection of p53 mutations in hepatocellular carcinoma (HCC) surgical resections using the FASAY (functional analysis of separated alleles of p53 on yeast) procedure.

METHODSp53 status was analyzed by FASAY and cDNA sequencing in 50 cases of HCC. After the extraction of RNA from the frozen tumor and corresponding normal tissues, reverse transcription RT-PCR was carried out using these samples. The assay can detect mutations of p53 mRNA between codons 67 and 347 by the DNA-binding activity of the protein and reveal them as red colonies.

RESULTSOf the 50 specimens, 29 (58%) were positive (mutant) by FASAY. Sequencing analysis confirmed that all 29 FASAY positive tumors harbored mutations, and that no mutations were detectable in any FASAY negative tumors. In 29 p53 mutations, 22 mutations were point missense mutation, 5 were deletions and 2 were splicing mutations. A novel splice mutation on splice donor of intron 6 was reported, which could produce two different mRNAs, respectively using the nearest upstream and downstream recessive splice donor sites.

CONCLUSIONFASAY is a sensitive method for detecting the various types of p53 mutations in HCC, suggesting that the yeast functional assay for the detection of p53 mutations may be essential for elucidating their clinical significance.

Alternative Splicing ; genetics ; Carcinoma, Hepatocellular ; genetics ; DNA Mutational Analysis ; methods ; Gene Frequency ; Genetic Testing ; Humans ; Liver Neoplasms ; genetics ; Mutation ; Reproducibility of Results ; Sensitivity and Specificity ; Tumor Suppressor Protein p53 ; genetics

The effects of a non-selective inhibitor of cyclo-oxygenase (COX) indomethacin, and exogenous prostaglandin E(2) (PGE(2)) on A(delta) units and C units in the saphenous nerve of diabetic hyperalgesic rats were studied. The results showed that the conduction velocity of A(delta) units and C units and their mechanical threshold in diabetic hyperalgesic rats were obviously decreased, and a small number of A(delta) units (4/24) and C units (2/18) produced increased spontaneous activities. Intraperitoneal injection of indomethacin in diabetic hyperalgesic rats significantly relieved mechanical hyperalgesia, and resulted in a decrease in spontaneous afferent activities of the A(delta) units and C units. Subcutaneous injection of exogenous PGE(2) into the diabetic hyperalgesic and control rats produced a significant decrease in mechanical threshold of the A(delta) units and C units, and elicited discharge from 3 A(delta) units (3/24) and 1 C unit (1/18) in diabetic hyperalgesic rats and from 2 A(delta) units (2/13) in control rats. The present data suggest that the synthesis and release of PGs are increased in diabetic neuropathy, PGs can sensitize and /or activate A(delta) units and C units and elicit hyperalgesia and allodynia in diabetic rats.
Afferent Pathways ; drug effects ; physiopathology ; Animals ; Diabetes Mellitus, Experimental ; physiopathology ; Femoral Nerve ; drug effects ; physiopathology ; Hyperalgesia ; physiopathology ; Indomethacin ; pharmacology ; Male ; Pain Threshold ; drug effects ; Prostaglandin Antagonists ; pharmacology ; Rats ; Rats, Sprague-Dawley

Objective To evaluate the clinical outcomes of patients with acute myocardial infarction(AMI)complicating cardiogenic shock underwent various treatments.Methods From January,2002 to May,2007,47 AMI patients with cardiogenic shock were treated in our department by optimal medication (dopamine,epinephrine,norepinephrine,etc.),intrar-aortic balloon pump(IABP),mechanical ventilation when indicated,percutaneous coronary intervention(PCI)or coronary artery bypass graft (CABG).Outcome and factors related to mortality for these patients were analyzed in this retrospective study.Results Besides optimal medication and IABP in all patients,31 patients underwent PCI(66.0%),6 patients received emergency CABG(12.8%).The overall in-hospital mortality rate was 36.2%(17/47),6 patients(14.9%)died before coronary revascularization and 11 patients(21.3%)died after revascularization.Nine patients died of pump failure and 8 patients died of renal and(or)respiratory failure.Regression analysis showed that acute renal failure(r=0.734,P=0.000),acute respiratory failure (r=0.606,P=0.000)and diabetes(r=0.372,P=0.012)were positively related to in-hospital mortality.Conclusion Despite improvements in treatment options for AMI patients complicating cardiogenic shock,in-hospital mortality remained high,especially for patients complicating further with acute renal failure and acute respiratory failure.

Objective To explore whether or not the glioma U251 stem cell regulates its resistance to TMZ by the FOXO3a/β-catenin pathway.Methods The U251 stem cells were divided into the TMZ group(100 μmol/IL TMZ treated cells)and the control group.The Western blot and real-time quantitative reverse transcription-PCR(qRT-PCR)were used to detect the expression levels of FOXO3a,β-catenin,Nestin,CD133 and Sox2 under the TMZ action in the two groups.The stem cell pelletization experiment was used to verify the resistance of U251 stem cell on TMZ.Recombinant viral vectors pHY-FOXO3a,pHY-β-catenin-KD and PHy-β-Catenin-KD were constructed by embedding FOXO3a/β-catenin interference sequences into lentivirus pHY-LV-KD1.1 expression vector and transfected into U251 stem cells.The change of stem cell clone pellets number was measured under TMZ action.The effects of knockdown FOXO3a and β-catenin on the characteris-tics and drug resistance of U251 stem cells were observed.Results The Western blot and qRT-PCR results showed that compared with the control group,the expression levels of FOXO3a,β-catenin,Nestin,CD133 and Sox2 protein in the TMZ group were increased,the Nestin,CD133,Sox2 mRNA expression levels were in-creased(P＜0.05).The clone formation experiment results showed that the majority of survival cells on 5 d after TMZ treatment were the stem cells,indicating that the U251 stem cells could better tolerate TMZ.Knoc-king down FOXO3a and β-catenin could reduce the U251 stem cell populations number,indicating that its re-sistance to TMZ was weakened.Conclusion The FOXO3a/β-catenin pathway could regulates the characteris-tics of U251 stem cell and TMZ resistance.