Objective To investigate the effect of bumetanide pretreatment on focal cerebral ischemiareperfusion(I/R) injury in rats.Methods One hundred and five male SD rats weighing 250-300 g were randomly divided into 3 groups (n =35 each ):sham operation group(group S),focal cerebral I/R group (group I/R) and bumetanide pretreatment group (group B).Focal cerebral I/R was induced by occluding the fight middle cerebral artery with a nylon thread with a rounded tip which was inserted into internal carotid artery and advanced cranically until resistance was met in groups I/R and B.In group B bumetanide 30 mg/kg was injected iv at 10 min before ischemia.Neurologic function was assessed and scored-neurologic deficit scores (0 =no deficit,4 =unable to move).The animals were sacrificed at 3,24 and 48 h of reperfusion and their brains were immediately removed for determination of cerebral water content and expression of Na+ -K+ -2Cl- cotransporter 1 (NKCC1).The infarct size was measured at 24 h of reperfusion.Results Focal cerebral I/R significantly increased neurelogic deficit scores,NKCC1 expression,cerebral water content and infarct size in group I/R as compared with group S.Bumetanide pretreatment significantly attenuated cerebral focal I/R-induced increase in neurologic deficit scores,NKCC1 expression and cerebral water content in group B as compared with group I/R.There was no significant difference in infarct size between groups I/R and B.Conclusion Bumetanide pretreatment can reduce focal cerebral I/R injury in rats,and down-regulation of NKCC1 expression is involved in the mechanism.

Objective To study the gene expression of Wnt signal transduction pathway in experimental liver fibrosis and to investigate its role in liver fibrosis. Methods Liver fibrosis model was induced with carbon tetrachloride in 8 SD rats. Another 8 healthy rats were served as control. The gene expression in liver tissues of models and controls were examined using real time PCR array. The differential gene expression was identified as either up- or down-regulated 2-fold. The expressions of smooth muscle actin (SMA), Wnt4, Frizzled2 and β-catenin in the tissues were examined by immunohistochemistry and Western blot. Results The examination confirmed that 36 genes were differentially expressed, including 25 genes up-regulated and 11 genes down-regulated. Compared with the controls, the expressions of Wnt4, Wnt5 a and W nt11 were up-regulated more than 13.9-, 16.5-and 2.17-fold respectively, while the expressions of Wntl and Wnt3 were down-regulated more than 2.32- and 2.15-fold respectively in fibrotic liver. Immunohistochemistry and Western blot showed that the expressions of SMA, Wnt4 and Frizzled2 in fibrotic liver were remarkably higher than those in normal controls. While the level of phosphorylated β-catenin was decreased. Conclusion Both canonical and noncanonical Wnt signal transduction pathway may involve in the mechanism of liver fibrogenesis.

Objective To estimate the value of neck circumference in diagnosis of metabolic syndrome. Methods We analyzed the neck circumference of 523 volunteers and the occurrence of metabolic syndrome among them. Then we updated and revalued their data after follow-up for 1.5 years, meanwhile, we evaulated the value of Body Mass Index (BMI) and neck circumference in predicting metabolic syndrome. Results The increase of neck circumference was closely related to the occurrence of metabolic syndrome , and this tendency continued in the follow-up. Compared with BMI , neck circumference was sensitive in identifying the patients of metabolic syndrome. Conclusion Neck circumference has great value in diagnosis and treatment of metabolic syndrome.

Objective To evaluate the effect of ulinastatin on the expression of aquaporin-4 (AQP4) during focal cerebral ischemia-reperfusion (I/R) in rats.Methods One hundred and thirty-five male adult SpragueDawley rats,weighing 230-280 g,were randomly divided into 3 groups (n =45 each):sham operation group (S group),I/R group and ulinastatin group (group U).The rats were anesthetized with intraperitoneal 10％ chloral hydrate 3.5 ml/kg.Focal cerebral ischemia was induced by 2 h middle cerebral artery occlusion followed by 24 repeffusion.In U group,ulinastatin 100000 U/kg was injected immediately after beginning of reperfusion,while the equal volume of normal saline was injected in S and I/R groups.The rats were sacrificed at 6,24 and 48 h of repeffusion and brains were removed for determination of infract volume (by TTC),brain water content and expression of AQP4 (by immunohistochemistry) in brain tissues.Results Compared with S group,the infarct volume and brain water content were significantly increased,and the expression of AQP4 was up-regulated at each time point in I/R and U groups (P ＜ 0.05).Compared with I/R group,the infarct volume and brain water content were significantly decreased,and the expression of AQP4 was down-regulated at each time point in U group (P ＜0.05).Conclusion The mechanism by which ulinastatin mitigates focal cerebral I/R injury is related to down-regulation of AQP4 expression in brain tissues.

AIM: To investigate the effect of autologous bone marrow mononuclear cells (BM-MNCs) local transplantation (BM-LT) on a rabbit model of hindlimb ischemia. METHODS: After unilateral femoral artery and its branches were surgically ligated and excised in rabbits (n=14) 1 hour or so, autologous BM-MNCs were injected into the ischemic skeletal muscles in 7 rabbits, and phosphate buffered saline for another 7 ones as control. Hemodynamics parameters were measured at day 0, day 7 and day 28, respectively. On day 28, the animals were sacrificed in groups for pathological analysis. RESULTS: Compared with control group, color Doppler flow image in BM-LT group was recovered significantly, and its' systolic peak velocity and end-diastolic velocity were also improved (P

OBJECTIVE To observe the regulation effect of chidamide on energy metabolism in HCT-8 and HT-29 cells. METHODS HCT-8 and HT-29 cells were treated with chidamide 5,10 and 20 μmol · L-1. Morphological changes of these cells were observed under an ordinary optical microscope. Cell proliferation was detected by MTT. ATP production was determined by CellTiter-Glo? assay kit. Metabolic changes were tested by glycolytic stress kit. The mRNA level of lactate dehydrogenase A (LDH-A)was analyzed by real-time quantitative PCR,whereas the protein level of LDH-A was analyzed by Western blotting. RESULTS Compared with control group,cell morphology of HCT-8 and HT-29 cells in chidamide treated group was irregular,accompanied by deformation,shrinkage and cell debris, and the inhibitory rate of proliferation increased(P<0.05). There was no significant difference in ATP total content between chidamide 5 and 10 μmol · L-1 16 h treatment groups,but in chidamide 20 μmol · L-1 treatment group it was decreased(P<0.05). Chidamide 20μmol · L-1 had no effect on oxygen consumption rate, but glycolysis ATP generation rate was reduced by 30.7% and 37.9%(P<0.05),respectively. Chidamide 20μmol · L-1 had no effect on LDH-A mRNA level,but it decreased the protein level of LDH-A(P<0.01). CONCLUSION Chidamide can abate the respiratory metabolic ability of HCT-8 and HT-29 cells. The mechanism may be related to the down-regulation of LDH-A.

Objective To verify enzyme activity inhibition of a novel histone deacetylase inhibitor ( HDACi ) JZ005 using an HDACi chemiluminescence detection kit and a cell-based screening model .Methods The plasmid with p21 gene promoter elements and luciferase reporter gene was transfected into human embryonic kidney cells 293 , and the stable transfectants were established by G418 screening.Enzyme activity inhibition of JZ005 on histone deacetylases (HDACs) was verified by the HDACi chemiluminescence detection kit and the cell-based screening model .A well-known HDACi , tri-chostatin A ( TSA) was used as the positive control .MTT assay was used to detect the protection of rat H 9c2 myocardial cells suffering from CoCl 2-induced hypoxia and treated with different concentrations of JZ 005 .The expression of acetylated histone H3 protein of normal and CoCl 2-induced hypoxia H9c2 cells before and after JZ005 treatment was assayed by West-ern blotting while the effect of drug administration on apoptosis was detected by flow cytometry ( FCM) .Results An HDA-Ci cell-based screening system targeting the p21 gene promoter was ranging established .The JZ005, a HDACi, markedly suppressed the activity of HDACs by more than 50%with the concentration ranging from 50 to 400 μmol/L.JZ005 signifi-cantly protected H9c2 cells from hypoxia injury .Cell viability was increased by 38.33%,56.00% and 35.20% compared with control,accompanied by an enhanced acetylation level of histone H 3.JZ005(25,50 and 100 μmol/L) treatment sig-nificantly decreased the number of apoptotic cells (6.63%,10.56% and 8.89%) compared to control group (12.89%). Conclusion An HDACi cell-based screening system is successfully established .JZ005 effectively protects myocardial cells against hypoxia injury while enhancing the acetylation level of histone H 3.Our results indicate that JZ005 might be developed as a potential drug for hypoxia treatment .

OBJECTIVETo study the effect of platelet-derived growth factor-BB (PDGFBBB) on rat corpus cavernosum smooth muscle (CCSM) cell proliferation, migration and phenotypic modulation and explore the underlying mechanisms.

METHODSWistar rat CCSM cells were obtained through a modified tissue culture method and identified by immunofluorescence assay. The effect of PDGFBB on the proliferation of CCSM cells was investigated using a CCK-8 kit and the optimum PDGFBB concentration for cell treatment was determined. CCSM cells were treated with vehicle or PDGF-BB at the optimum concentration, and the cell migration was examined using scratch assay; the mRNA expression of the transcription factor myocardin and the contractile phenotype markers αSMA and SMMHC in CCSM cells were determined by qRT-PCR at 24 h and 48 h. The protein expression of myocardin in CCSM cells incubated with PDGFBB for 0, 24 and 48 h was examined by Western blotting.

RESULTIn CCSM cell culture, 96.5%and 96% of the cells were positive for αSMA and smoothelin, respectively. PDGFBB at different concentrations markedly promoted the proliferation of CCSM cells; the optimum PDGFBB concentration for enhancing cell proliferation was 12.5 ng/mL, which induced the migration of CCSM cells and significantly reduced the mRNA expressions of myocardin, αSMA and SMMHC (P<0.01). Exposure to PDGFBB decreased the protein expression of myocardin as the exposure time extended (within 48 h).

CONCLUSIONCCSM cells of a high purity can be obtained by the modified tissue culture method. PDGFBB can promote the proliferation and migration of CCSM cells and cause a phenotypic conversion from the contractile to the synthetic type possibly by down-regulating myocardin.

Actins ; metabolism ; Animals ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Down-Regulation ; Male ; Myocytes, Smooth Muscle ; cytology ; drug effects ; Myosin Heavy Chains ; metabolism ; Nuclear Proteins ; metabolism ; Penis ; cytology ; Phenotype ; Proto-Oncogene Proteins c-sis ; pharmacology ; RNA, Messenger ; Rats ; Rats, Wistar ; Trans-Activators ; metabolism

Objective To investigate the relationship between hypoxia-related genes and ischemic stroke,and to screen for molecular markers related to hypoxia. Methods First,GSE16561 and GSE58294 datasets and hypoxia-related gene sets were downloaded to screen for hypoxia-related differentially expressed genes. After functional annotation,these hypoxia-related differentially expressed genes were analyzed by the least absolute shrinkage and selection operator least absolute shrinkage and selection operator(LASSO) regression and support vector machine algorithm to screen for immune-related biomarkers,and differential immune cells were analyzed between the ischemic stroke group and the control group. Finally,transcription factors and miRNAs were predicted. Results A total of 13 hypoxia-related differentially expressed genes were screened,which were enriched mainly in the processes of glucose transmembrane transport,monosaccharide transmembrane transport,and mitophagy regulation. Based on the LASSO regression and support vector machine algorithm analyses,eight hypoxia-related biomarkers were identified,which were significantly correlated with most immune cells. Furthermore,the network of 209 transcription factors corresponding to 8 genes and 25 miRNAs corresponding to 6 genes were predicted. Conclusion Eight hypoxia-related biomarkers are identified,which may be used as potential therapeutic targets for inhibiting brain injury after stroke. In addition,glucose metabolism and immune mechanisms may play an important role in the occurrence and progression of ischemic stroke.

The present study was performed to investigate the seroprevalence and risk factors for Dirofilaria immitis infection in cats from Liaoning province, northeastern China. From October 2014 to September 2016, sera of 651 cats, including 364 domestic cats and 287 feral cats (332 females and 319 males) were assessed. They were tested for the presence of D. immitis antigen using SNAP Heartworm RT test kit. In this population, the average prevalence was 4.5%. Age and rearing conditions (feral or domestic) were found to be associated with the prevalence of D. immitis. The prevalence was significantly higher in feral cats compared with domestic cats (8.4% vs 1.4%, P < 0.01). There was no significant difference between males and females (4.7% vs 4.2%, P>0.05), but older cats (≥3 years old) showed a statistically higher prevalence compared with younger cats ( < 3 years old) in feral populations (16.8 vs 2.4%, P < 0.01), while the difference between the age groups was not statistically significant in domestic cats (2.4% vs 0.51%, P>0.05), all these results suggest that outdoor exposure time may be one of the most important factors for D. immitis prevalence in cats. Results reveal that D. immitis are prevalence in domestic and feral cats in northeastern China, which indicates that appropriate preventive measures should be taken to decrease the incidence of feline heartworm disease in Liaoning province, northeastern China.
Animals ; Cats* ; China* ; Dirofilaria immitis* ; Dirofilaria* ; Dirofilariasis ; Female ; Humans ; Incidence ; Male ; Prevalence ; Risk Factors ; Seroepidemiologic Studies*