Toxicarioside A inhibits SGC-7901 proliferation,migration and invasion via NF-κB/bFGF signaling

AIM:To investigate the inhibitory role of toxicarioside A on the gastric cancer cell line human gastric cancer cell line(SGC-7901) and determine the underlying molecular mechanism.METHODS:After SGC-7901 cells were treated with toxicarioside A at various concentrations(0.5,1.5,4.5,9.0 μg/mL) for 24 h or 48 h,cell viability was determined by 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl2H-tetrazolium bromide assay,and the motility and invasion of tumor cells were assessed by the Transwell chamber assay.Immunofluorescence staining,reverse transcription polymerase chain reaction and Western blotting were performed to detect the expression of basic fibroblast growth factor(bFGF) and fibroblast growth factor receptor-1(FGFR1),and nuclear factorkappa B(NF-κB) activation was examined by electrophoretic mobility shift assay.RESULTS:The results showed that toxicarioside A was capable of reducing cell viability,inhibiting cell growth,and suppressing cell migration and invasion activities in a time-and dose-dependent manner in SGC-7901 cells.Further analysis revealed that not only the expression of bFGF and its high-affinity receptor FGFR1 but also the NF-κB-DNA binding activity were effectively blocked by toxicarioside A in a dose-dependent manner compared with the control group(P < 0.05 or P < 0.01).Interestingly,application of the NF-κB specific inhibitor,pyrrolidinedithiocarbamate(PDTC),to SGC-7901 cells significantly potentized the toxicarioside A-induced down-regulation of bFGF compared with the control group(P < 0.05).CONCLUSION:These findings suggest that toxicarioside A has an anti-gastric cancer activity and this effect may be achieved partly through down-regulation of NF-κB and bFGF/FGFR1 signaling.

Effect of hypoxia/reoxygenation on the viability expression of ROS and MAPKs in myocardial cells

Objective:To investigate to the expression effect of hypoxia and hypoxia/reoxygenation on ROS,MAPKs and cell apoptosis in H9c2 cardiomyocytes.Methods:H9c2 cells were treated with cobalt chloride(CoCl2)to establish the chemical hypoxia and hypoxia/reoxygenation-induced cardiomyocyte injury model.CoCl2 was used to process cells at different concentrations from 150-2400μmol/L and different time from4-24 h;H9c2 cells viability was detected by MTT,and the intracellular ROS level was measured by 2’,7’-dichlorflμoresceindiacetate(DCFH-DA)and dihydroethidiμm(DHE)staining and photoflurography.The active expression level of mitogen-activated protein kinases(MAPKs)(including JNK,ERK and p38)and caspase-3.Results:At the concentration from 300 to 1200μmol/L,CoCl2 does/time-dependently inhibited the cell viability in H9c2 cells(P<0.01).Compared with control group,the ROS levels in hypoxia group were significantly increased(P<0.05).In hypoxia group,the active expression levels of p-JNK,p-p38 and caspase-3 was higher than those in control group(P<0.05).However,the expression of p-ERK wasn’t significant differernce.Furthermore,all the expression levels of ROS,p-JNK,p-ERK,p-p38 and caspase-3 in H/R group were significantly raised compared with hypoxia group(P<0.01).Conclusions:Reoxygenation further aggravate chemical hypoxia induced cardiomyocyte oxidative stress injury by activating ROS/MAPKs signals,suggesting the role of myocardial ischemia/reperfusion injury in the pathogenesis of ischemic heart disease.

Relationship between fever and early clinical diagnosis and treatment in mild COVID-2019 patients

Objective:To investigate the relationship between clinical manifestations,blood testing,chest CT,treatment and fever of 457mild cases of COVID-19.Methods:A tatal of 457cases of mild COVID-19 diagnosed in Wuhan Jianghan module Hospital from February 6,2020 to February 26,2020 were gathered,and the cases with/without fever were retrospectively analyzed in epidemiology,clinical manifestation,blood routing,hypersensitive C-reactive protein(hs-CRP),pathogens serological testing,chest CT and drugs treatment.Results:For 312 fever cases,the average age was(47.6±10.9)years old,male was 128 cases(41%),the mean peak fever was 38(36.8,38.6)℃,fever lasting was 2(0,3)days,131cases(28.7%)had Huanan seafood market exposure history,90 cases(28.8%)had closely contact with COVID-19 patients,20cases(6.4%)had hypertension history,10 cases(3.2%)had diabetes,5cases(1.6%)had coronary heart disease.Main symptoms of fever group including dry cough of 260cases(83.3%),of 104cases(33.3%),blood sputum of 9cases(2.9%),chest tightness 20cases(19.6%),shortness of breath of 87cases(27.9%),weak of 105cases(33.7%),diarrhea of 43cases(13.8%),sleep disorders of 61cases(19.6%),among fever cases with sputum and shortness of breath more rather without fever,and two groups were statistically significantly difference(P<0.05).In fever group,WBC(109/L)[4.34(3.69,5.26)than 5.11(4.22,6.22),P<0.001],NEUT(109/L)[2.67(2.1,3.49)than 2.88(2.37,3.9),P<0.05],LYM(109/L)[0.9(0.8,1.1)than 1.36(1.03,1.85),P<0.001),PLT(109/L)/(106,188)134-189(132,224),P<0.001)were lower than normal group,while WBC<4.0×109/L[108(34.6%)than 24(16.6%),P<0.001)],LYM<1.0×109/L[189(60.6%)than 31(21.4%),P<0.001)],PLT<100×109/L[45(14.4%)than in 2(1.4%),P<0.001)and hs-CRP(mg/L)[14(6.83,32)than 3.4(0.96,10.75),P<0.001)were higher than normal group,and pathogens serological examination of 36cases shown influenzaⅠ/Ⅱvirus IgM antibody positive in 3cases(8.3%),respiratory syncytial virus IgM antibody positive in 2 cases(5.6%),mycoplasma pneumoniae IgM antibody positive(11.1%),4 cases of adenovirus IgM antibody positive(11.1%),but 4 pathogens of fever group and normal group were no statistical difference(P>0.05).According to early onset of chest CT examination,there were no change of 11cases(2.4%),unilateral lung lesions of 93cases(20.4%),bilateral lung lesions of 353 cases(77.2%),ground glass shadow of 228 cases(73.1%),high density shadow patch of 65 cases(20.8%),consolidation shadow of 6 cases(1.9%),while fever group compared with normal group,there were no statistically significant difference(P>0.05).Logistic regression analysis showed that LYM(OR=0.24,95%CI=0.139~0.415,P<0.001),LYM<1.0 x 109/L(OR=5.12,95%CI=3.07~8.65,P<0.001),PLT<100×109/L(OR=6.1,95%CI=1.36~27.33,P<0.05),and the hs-CRP(OR=1.03,95%CI=1.02~1.05,P<0.001)were fever independent impact factors in mild COVID-19.Aiming at COVID-19 treatment,Chinese medicine usage had 416cases(91%),antiviral drugs had 328cases(71.8%),and antibiotics had 172cases(37.6%),furthermore,fever group using antiviral and antibiotic drugs was higher than normal group(P<0.05),and compared to low fever group,high fever group used more antibiotic drugs(P<0.05).Conclusions:Mild COVID-19 patients with early onset had epidemiological characteristics,and fever,dry cough,expectoration and shortness of breath were main symptoms,and chest CT often involved bilateral lung lesions,ground glass shadow and patch density shadow.The fever patients in WBC,LYM,PLT,hs-CRP index changed significantly and could affect on anti-infective therapy selection.

Effects of high glucose on expression of OPG and RANKL in rat aortic vascular smooth muscle cells

Objective:To explore effect of high glucose on expression of osteoprotegerin(OPG) and receptor activator of NF- κB ligand(RANKL) in rat aortic vascular smooth muscle cells.Methods:SD rats were intraperitoneally injected with streptozotocin,OPG and RANKL expression in rat thoracic aortas were detected by immunohistochemical staining.In cultured vascular smooth muscle cells(VSMCs)(A7r5),qRT-PCR and Western blot analysis were used to examine the mRNA and protein levels of OPG and RANKL.Results:Our results demonstrated that OPG expression was increased in hyperglycemic rat aortic VSMCs.while RANKL expression was decreased.Besides,in vitro experiments high glucose induced OPG expression,but depressed RANKL expression by dose- and time-dependent manner in cultured A7r5.Conclusions:Our findings suggested that high glucose could promote the expression of OPG,and inhibit the expression of RANKL in VSMCs,which may be partly be the molecular mechanism of diabetic vascular calcification.

p42/p44 mitogen-activated protein kinases inhibit atrial natriuretic peptide mRNA transcription in gp130-mediated hypertrophic ventricular myocytes

Objective:To understand the role of ANP mRNA transcription regulation in gpl30-mediated cardiomyocyte hypertrophy,and the involved mitogen-aetivated protein kinase kinase(MEK)-extracellular signal-regulated kinase(ERK,also called p42/p44 MAPK)signaling pathway.Methods:isolated neonatal ventricular myocytes were treated with different concentrations of CT-1(10^(-9),10^(-8)and 10^(-7)mol/L).MTT was used to analyze the viability and RT-PCR was used to detect ANP mRNA levels in eardiomyocyte.To inhibit p42/p44 MAPK activity in hypertrophic cardiomyoeytes,the cells were pretreated with a specific MEKI inhibitor.Results:CT-1significantly induced ANP mRNA expression and the viability of canliomyocytes in a doseand time-dependent manner.Furthermore,blocking p42/p44 MAPK activity by the special MEk1 inhibitor uprcgulatcd the ANP mKNA.Conclusions:p42/p44 MAPK have an important role in suppressing ANP mRNA transcription and cell activity in gpl30-mediated hypertrophic ventricular myocytes.

Comparison of the curative effect of intracoronary retrograde thrombolysis and thrombus aspiration combined with stent implantation on STEMI patients

Objective:To evaluate the short-term and long-term curative effects of precise intracoronary retrograde thrombolysis combined with stent implantation,thrombus aspiration combined with stent implantation,and traditional stent implantation in patients with acute ST-segment elevation myocardial infarction.Methods:From January 2018 to October 2019,184 patients diagnosed with acute ST-segment elevation myocardial infarction and infarction-related arterial blood flow TIMI 0 grade in the First Affiliated Hospital of Hainan Medical College and percutaneous coronary intervention(PCI)were selected.According to different surgical methods,patients were divided into intracoronary retrograde thrombolysis combined with stent implantation group(thrombolysis group,n=57 cases),thrombus aspiration combined with stent implantation group(aspiration group,n=57 cases)),traditional stent implantation group(traditional group,n=70 cases).Compare the incidence of no-reflow phenomenon after percutaneous coronary intervention,the rate of 1 hour ST_segment fall≥50%in the ECG after PCI,and the main adverse cardiovascular events(MACE)during hospitalization;compare the left ventricular end-diastolic diameter(LVEDD)、left ventricular ejection fraction(LVEF)and major adverse cardiovascular events at 1 year after PCI.Results:1.The short-term effects:The incidence of no-reflow phenomenon in the thrombolytic group was lower than that of the aspiration group and the traditional group,and the rate of 1hSTR≥50%was higher than that of the aspiration group and the traditional group,the difference was statistically significant(P<0.05).2.The long-term effects:1 year after percutaneous coronary intervention,the LVEDD of the thrombolytic group was lower than that of the aspiration group and the traditional group,while the LVEF was higher than the aspiration group and the traditional group,and the incidence of major adverse cardiovascular events in the thrombolytic group at was lowest,both the difference was statistically significant(P<0.05).Conclusion:The application of intracoronary retrograde thrombolysis combined with stent implantation in STEMI patients can reduce the occurrence of no-reflow,improve long-term cardiac function,and reduce the occurrence of major adverse cardiovascular events for short-term and long-term.