Inhibitory effect of tumor suppressor p33^(ING1b) and its synergy with p53 gene in hepatocellular carcinoma

AIM: To investigate the inhibitory effect of tumor suppressor p33ING1b and its synergy with p53 gene in hepatocellular carcinoma (HCC).METHODS: Recombinant sense and antisense p33ING1b plasmids were transfected into hepatoma cell line HepG2 with lipofectamine. Apoptosis, G0/G1 arrest, cell growth rate and cloning efficiency in soft agar of HepG2 were analyzed after transfection. In three hepatoma cell lineswith different endogenous p53 gene expressions, the synergistic effect of p33ING1b with p53 was analyzed by flow cytometry and luciferase assay was performed to detect the activation of p53 downstream gene p21WAF1/CIP1. In addition, the expression and mutation rates of p33ING1b in HCC tissues were measured by immunohistochemistry and polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP).RESULTS: Overexpression of p33ING1b inhibited cell growth of HepG2, induced more apoptosis and protected cells from growth in soft agar. Combined transfer of p33ING1b and p53 gene promoted hepatoma cell apoptosis, G0/G1 arrest and elevated expression of p21WAF1/CIP1. Immunostaining results showed co-localized P33ING1b with P53 protein in HCC tissues and there was a significant relation between protein expression rates of these two genes (P＜0.01).Among 28 HCC samples, p33ING1b presented a low gene mutation rate (7.1%).CONCLUSION: p33ING1b collaborates with p53 in cell growth inhibition, cell cycle arrest and apoptosis in HCC. Loss or inactivation of p33ING1b normal function may be an important mechanism for the development of HCC retaining wildtype p53.

Antiangiogenic effect of berberineα-hydroxy-δ-decanoylethyl sulfonate on Lewis lung carcinoma

OBJECTIVE To investigate the antiangiogenic and antitumor effects of berberineα-hydroxy-δ-decanoylethyl sulfonate(HB)in vitro and in vivo.METHODS MTT assay was employed to determine the proliferation of tumor cells.Western blot analysis was used to detect the expression of VEGF and MMP-9.Transwell co-culture was used to determine the cell migration at the conditioned medium of Lewis lung carcinoma cells exposed to HB for 24 h.The C57BL/6 mice bearing Lewis lung carcinomas were treated with vehicle,different doses of HB(60,90 and 120mg·kg-1,ig)for 20 d,or cyclophosphamide(50mg·kg-1,ig)at d 1,d 8 and d 15.Afterwards,the xenografted tumors were exercised and weighed,and the lung metastasis was determined by HE stainingof the lung tissues.Tumor angiogenesis was determined by CD34 staining and microvessel density(MVD)by immunohistochemistry.RESULTSHB inhibited the growth of Lewis lung carcinoma cells in a time-dependent manner with IC50 value of 8.87,2.26 and 0.98mg·L-1,respectively when the cells were treated with HB for 24,48 and 72h.Cell migration induced by the conditioned medium of Lewis lung carcinoma cells treated with 2.5,5 and 10mg·L-1 HB was reduced by 37%,54% and 63%respectively(P<0.01).HB suppressed the expression of angiogenesis factors in a concentration-dependent manner.HB was effective in inhibition of MMP-9 expression at 5,10 and 20mg·L-1(P<0.05),whereas HB 20mg·L-1 was minimal effective concentration in inhibition of VEGF expression(P<0.01).On C57BL/6 mice bearing Lewis lung carcinomas,HB significantly reduced the tumor burden and MVD in tumor mass as well as inhibited lung metastasis at doses of 90 and 120mg·kg-1.CONCLUSION HB produces reliable antiangiogenic effect,inhibits the growth and metastasis of Lewis lung carcinoma.These effects may be attributed to its ability of suppressing the expression of VEGF and MMP-9,and reducing MVD.

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.

RADIAL PROFILE OF THE SOLID FRACTION IN A LIQUID-SOLID CIRCULATING FLUIDIZED BED

In a liquid-solid circulating fluidized bed, lateral forces acting on the particles determine the movement of the particles in the radial direction, and this creates a radial profile for the solid fraction. This work proposes a model to calculate the radial profile of the solid fraction in a liquid-solid circulating fluidized bed based on the balance of the lateral force and the turbulent dispersion force.