Child ; Humans ; Intestinal Polyps ; genetics ; Molecular Biology

Child ; Gastroesophageal Reflux ; diagnosis ; therapy ; Humans

Objective To investigate the effects of extracellular-signal regulated kinase (ERK) on hypoxic-ischemic brain damage of neonatal rats.Methods The hypoxic-ischemic brain damage model of neonatal Wistar rats was established as following:first the right common carotid artery of the rats was ligated;2 h after operation,the rats began to inhale 8%-oxygen oxygen-nitrogen gas mixture lasting for 2 h.Rats were randomly divided into three groups:sham-group,hypoxic-ischemic group and ERK inhibitor PD98059 group (the rats were injected PD98059 2 mg/kg 10 min before the ligation).Six hours after the models were done,hippocampi and cortex of the ligation side of rats in the three groups were collected,and the levels of malondialdehyde (MDA) and superoxide dismutase (SOD) were measured.Apoptosis of neuron was assessed by TUNEL staining.The expression of ERK1,ERK 2,Bcl-2 and Bax were examined by Western blot.The differences among the groups were analyzed with ANOVA and q test.Results Compared with the sham-group,the MDA level [(342.9± 10.8) μmol/L vs (181.5± 17.0) μmol/L,q= 6.35,P＜0.01) and the apoptosis rate of neuron [(18.80±1.37)% vs (3.53±0.34)%,q=6.06,P＜0.01) of hypoxic-ischemic group was higher,and SOD level was lower [(34.8±4.3) U/ml vs (63.4±4.3) U/ml,q=4.99,P＜0.01].While the apoptosis rate of neuron [(15.53±0.64) %] and MDA level [(252.0± 17.1) μmol/L] of PD98059 group were lower than those of hypoxic-ischemic group(q=3.87 and 5.28,P＜0.01respectively),the SOD level [(51.5 ± 3.8) U/ml] was higher than that of hypoxic-ischemic group (q=4.17,P＜0.01).There were no differences of ERK1 and ERK2 expressions among the three groups.The phosphorylated ERK1 and ERK2 levels of hypoxic-ischemic group were higher than those of sham-group (q=3.82 and 4.08,P＜0.01) and PD98059 group (q=4.79 and 5.12,P＜0.01).The expression of Bcl-2 and Bax of hypoxic-ischemic group were higher than those of sham-group (q=3.55 and 3.42,P＜0.01).Compared with hypoxic-ischemic group,Bcl-2 expression (q=3.71,P＜0.01) of PD98059 group was higher,and Bax expression (q=5.86,P ＜ 0.01) was lower.Conclusions ERK is involved in hypoxic-ischemic brain damage of neonatal rats through regulating the expression of apoptosis protein.

Objective:To investigate the role of recombinant E.coli LLO/OVA on regulating the function of murine CD4+ CD25+ Treg cells.Methods:After E.coli LLO/OVA or E.coli OVA vaccination,the murine spleen CD4+ CD25+ Treg,CD4+ CD25- T and CD11c cells were collected respectively by magnetic beads sorting.The concentration of IL-10 in the supernatant of mix cocultured CD4+ CD25+ Treg and CD11c cells,and the suppression role of CD4+ CD25+ Treg cells on the proliferation of CD4+ CD25- T cells were determined.The percentage of OVA specific CD8+ T cells in mouse spleen was analyzed by flow cytometry.The number of metastatic tumor nodules in lungs of the mice transplanted with B16-OVA subcutaneouly was compared before and after dilition of CD4+ CD25+ Treg cells in mice.Results:Compared to E.coli OVA,E.coli LLO/OVA significantly downregulated IL-10 secretion of CD4+CD25+Treg cells and attenuated the suppressive effect of CD4+ CD25+ Treg on the proliferation of CD4+ CD25- T cells(P

Objective To develop a HPLC-UV method for the determination of [6]-gingerol in the rat plasma.Methods [6]-Gingerol was extracted from the rat plasma by using liquid-liquid extraction,then was separated on a Hypersil  C18 column(250 mm?4.6 mm,5 ?m).The mobile phase was acetonitrile and water(45:55,v/v) with a flow-rate of 1.0 mL/min.The eluate from the HPLC column was monitored by the spectrophotometric detector at 280 nm.The injection volume was 20 ?L.[6]-Gingerol was identified based on its retention time compared with the reference standard.Results The linear calibration curve was obtained in the concentration range of 0.25～5.0 ?g/mL.The low quantitative limit was 0.1 ?g /mL.The RSD of inter-day was

Objective:To compare the shear bonding strength between two kinds of veneering ceramics respectively bonded to Vita Mark II.Methods:Vita Mark II samples were prepaired in the size of 8 mm ×6 mm ×2 mm.Veneering ceramic VM9 and Titankeram-ik were sintered on the Vita Mark II blocks respectively(n =1 0).The shear bond strength of each specimen was measured,the failure mode of the fracture surface was observed by SEM.The element in the bonding interface was analysed by field emission scanning elec-tron microscope(FE-SEM).Results:The values of shear bond strength (MPa)of VM9 and Titankeramik groups were 36.1 7 ±8.91 and 42.07 ±8.31 respectively(P >0.05).SEM observation showed tight bond between the two materials,FE-SEM results showed penetration of Al element from MarkⅡblock into veneering porcelain VM9 or Titankeramik.Conclusion:Veneered porcelain of VM9 and Titankeramik can be used as the individual veneering ceramics for Vita Mark II blocs.

ObjectiveTo observe the influence of calcium - sensing receptor (CaSR) on Fas/Fas ligand (Fas L) pathway during anoxia/reoxygenation (A/R)- induced cardiomyocytes apoptosis in neonatal rat. MethodsSingle cells were dissociated from minced hearts of 2 - day - old Wistar rats with a 0. 25％ solution of crude trypsin and then cultured as monolayers at a density of 5 x 104cells/cm2 in DMEM medium equilibrated with humidified air containing 5％ CO2 at 37C. Three days after the cells were seeded, the cultured cardiomyocytes were randomly divided into three groups. ( 1 ) control group: cardiomyocytes were continuously cultured for 26 hours in DMEM medium.(2) A/R group: cardiomyocytes underwent anoxia for 2 hours and reoxygenation for 24 hours.(3) GdCl3 group: 300μmol/L GdCl3 was added to the culture medium at the beginning of reoxygenation. Apoptosis of cardiomyocytes was assessed by flow cytometer and morphological alterations were observed with transmission electron microscope.The expression of CaSR, Fas and Fas L were analyzed by Western blot.Results The result of flow cytometer showed that cardiomyocytes apoptosis was 12. 18％ ± 1.54％ in A/R group,and was higher than that in the control group (P ＜0. 01 ). At the same time, mitochondrial cristae dissolution and disappearance could be detected. Compared with A/R group, GdCl3, a specific activator of CaSR, further enhanced cardiomyocytes apoptosis to 20. 25％ + 2. 87％ ( P ＜ 0. 01 ), along with an increment in CaSR, Fas and Fas L expressions ( P ＜ 0. 05 ). ConclusionCaSR is closely involved in cardiomyocytes apoptosia during anoxia/reoxygenation. CaSR could induce apoptosis of neonatal rat cardiomyocytes through Fas/Fas L receptor pathway.

Objective:To optimize the extraction process for the total alkaloids of Longzuan Tongbi Fang by central composite design and response surface method.Methods:The independent variables were the ethanol contention,solvent consumption and extraction time,and the dependent variable was the weighted overall desirability of the content of total alkaloids and kukoline.The extraction process for the total alkaloids of Longzuan Tongbi Fang was optimized by central composite design and response surface method.Results:The optimum extraction conditions were as follows:9-fold amount of solvent was used,refluxed for 130 min,extracted twice,and the ethanol concentration was 75%.Conclusion:The central composite design and response surface method used for the extraction optimization for the total alkaloids of Longzuan Tongbi Fang is very simple and highly predictive.

Objective To investigate the in vitro effect of wild-type PTEN gene transfection on the growth of human colon cancer cell lines. Methods With liposome transduction technique,the retrovirus vector pBp-PTEN,which contains wild-type PTEN gene segment and pBabe-puro,was transfected into Lovo,one of the PTEN-deficient human colorectal carcinoma cell lines.After identification by Western blotting,cell growth,cell cycle and apoptosis before and after transfection were studied. Results It was indicated by the cell growth curve that after transfection the curve of the transfection group exhibited a mild tendency with no obvious logarithmic growth phase,and the growth velocity was significantly lower than that of the control group(P