This article was aimed to investigate the cell proliferation , cell apoptosis and its related molecular mechanisms of the human gastric carcinoma cell line BGC-823 in v itro after treatment with cinnamaldehyde . The MTT Assay demonstrated the inhibitory effect of cinnamaldehyde . And the Flow Cytometry was used to determine its induction of cell apoptosis. The Hoechst 33342 was used to observe morphological changes during apoptosis . Moreover , quantitative real time PCR and western blot analysis were used to detect the effect of cinnamaldehyde on human gastric carcinoma cell line BGC-823 . The results showed that compared with the control group , cinnamaldehyde had inhibitory effect on human gastric carcinoma cell line BGC-823 ( P <0 . 01 ) . It showed that cinnamaldehyde induced apoptosis through the downregulation of Bcl-2 , Bcl-xL and Survivin expression , upregulation of Bax and Bak expression , downregulation of Bcl-2 and Procaspase-3 , and upregulation of BAX . It was concluded that cinnamaldehyde had inhibitory effect on the proliferation of human gastric carcinoma cell line BGC-823 and induced apoptosis . It may be related to the activation of the endogenous apoptosis pathway .

Objective To characterize the dimerization and the antigenicity of the ORF2 polypep-tide of hepatitis E virus (HEV, genotype 4). Methods HEV ORF2 gene was cloned from the serum of a patient with hepatitis E. The genotype was determined by sequencing. Three ORF2 polypeptides differing in size and other polypeptides with point mutations were produced in E. coli. The recombinant polypeptides were purified and analyzed by SDS-PAGE and Western blot. Results The ORF2 polypeptide containing 459-607 amino acid formed homedimer even in 8 mol/L urea. The truncated polypeptides containing amino acid 472-607 or 459-594 formed monomer only. The mutations at amino acid 562 or 595 disrupted the ho-modimer, whereas the mutations at amino acid 476 or 580 did not. Anti-HEV from hepatitis E patients only reacted with the homodimer form of the polypeptide 459-607 and did not react with monomer or tnmcated pol-ypeptides. Conclusion The amino acid 459-607 of HEV ORF2 is essential for dimerization of the ORF2 polypeptide. Residues at amino acid 562 and 595 are critical for the dimerization. The antigenicity of the polypeptide 459-607 mainly depends on its homodimer form.

Due to the good tumor-targeting and excellent biocompatibility, the drug-loading nanoparticles (NPs) has been widely applied in the diagnosis and treatment of cancer. However, after the NPs are recognized and internalized by cancer cells, the effects of NPs on cell migration behavior were unclear. In the present study, the self-assembly techniques (SAMs) was used to modify gold (Au) nanoparticles (Au NPs) with different chemical functional groups (CH3, OH, COOH and NH2) as model NPs. The dispersion of these groups in solution and the distribution in cells were studied by transmission electron microscope (TEM), respectively, and the proliferation was examined by MTT assay in vitro. The wound-healing and the Transwell assay were used to examine the effect of internalized Au-NPs on HepG2 cells migration. The results showed that different Au-NPs mainly distributed at the edge of the vesicle membrane and the gap between cells. The Au-NPs resulted in decreased cell viability in a concentration-depended manner. In addition, the results of wound-healing and Transwells assay indicated that the internalization of the NH2-NPs and OH-NPs would inhibit cell migration compared with those in the control group.
Carcinoma, Hepatocellular ; metabolism ; Cell Movement ; Cell Proliferation ; Cell Survival ; Gold ; Hep G2 Cells ; Humans ; Liver Neoplasms ; metabolism ; Metal Nanoparticles ; chemistry

This paper is aimed to investigate the effect of fluid shear stress on the tight junction of laryngeal squamous carcinoma (Hep2) cells and to explore the potential molecular mechanism. Hep2 cells were selected and subjected to the fluid shear stress of 1.4 dyn/cm2 for different time, respectively. The morphological changes of Hep2 cells under shear stress were observed using inverted microscope. The cell-cell junctions were examined by transmission electron microscope (TEM). The expressions of tight junction proteins (including Occludin, Claudin-5 and ZO-1) and the distribution of Claudin-5 were examined by Western blot assay and laser scanning confocal microscope, respectively. The results indicated that Hep2 cells turned to spindle-like shapes after exposed to shear stress, and showed the trend of the recovering to original shapes when the shear stress was cancelled. The cell-cell junctions were tight under the shear flow condition, and the permeability was reduced under the condition of 1.4 dyn/cm shear flow. The expressions of tight junction proteins were enhanced with increased duration of shear flow, but reduced after removing shear flow. The result of Claudin-5 expression by immufluorescence assay was consistent with that by Western blot. The Claudin-5 mainly distributed in the cytoplasm under static condition, while it located at the intercellular after shear flow stimulation, and it appeared intercellular and cytoplasm after stopping shear flow stimulation. Therefore, it can be concluded that shear stress changes the morphology of laryngeal squamous carcinoma Hep2 cells, and upregulates the tight junction.
Blotting, Western ; Carcinoma, Squamous Cell ; pathology ; Claudin-5 ; metabolism ; Hep G2 Cells ; Humans ; Laryngeal Neoplasms ; pathology ; Occludin ; metabolism ; Stress, Mechanical ; Tight Junctions ; Zonula Occludens-1 Protein ; metabolism

Objective To identify the imbalance of T cell-specific transcription factors T-bet/GATA-3,and to explore the modulation with dexamethasone and imiquimod in CD4+T cells from ovalbumin (OVA)sensitized mice.Methods CD4+T cells were obtained fromsingled-cell suspension of spleen(after lysis of RBC).ELISA assay was used to detect the concentrations of IL-4,IL-5 and IFN-γin superna tants and cell pellets,and the expression of T-bet and GATA-3 was detected by Western blot.Resuits In the control group,tIle low levels of IFN-γ were detected in the supernatants during 24 h.In OVA treatment group,the concentrations of IL-4,IL-5 were increased significantly,and the concentrations of IFN-γ were always low in the supernatants.In the dexamethasone treatment group,the concentrations of IFN-γ,IL-4 and IL-5 were all low in the supernatants during 24 h.In the imiquimod treatment group,the concentrations of IFN-γ were increased significantly,and the concentrations of IL-4 and IL-5 were decreased in the super natants.It worked at 6 h,and achieved the peak at 12 h,lasted over 24 h.In the control group,the expres sions of T-bet and GATA-3 were detected in CD4+T cells during 24 h.In OVA treatment group,the expressions of T-bet were decreased,and that of GATA-3 were increased rapidly in CD4+T cells.In dexam ethasone treatment group,the expressions of T-bet were always low in CD4+T cells,and that ofGATA-3 were no change during 24 h.In imiquimod treatment group,the expressions of T-bet were increased,andthat of GATA-3 were decreased in CD4+T cells.The protein expressions worked at 6 h.and achieved the peak at 12 h,lasted over 24 h.Conclusion The imbalance T cell-specific transcription factors T-bet/GA-TA-3 contributes to both high expression of GATA-3 and low expression of T-bet in CD4+T cells from OVA sensitized mice.Dexamethasone treatment inhibits the expression of T-bet in CD4+T cells and has no func tion in GATA-3.Imiquimod treatment modulates key master switches GATA-3 and T-bet that results in com mitting T helper cell to a TH 1 phenotype and imiquimod may play a key role in the regulation of TH2 cytokine responses in asthma.

To study the potential molecular mechanism of tumor angiogenesis in its microenvironment, we investigated the effects of HepG2 conditioned medium on the proliferation of vascular endothelial cell and vascular angiogenesis in our laboratory. Human umbilical vein endothelial EA. hy926 cells were co-cultured with HepG2 conditioned medium in vitro. The proliferation and the tubulogenesis of EA. hy926 cells were detected by teramethylazo salt azole (MTT) and tube formation assay, respectively. The results showed that the survival rate of the EA. hy926 cells was significantly increased under the co-culture condition. HepG2 conditioned medium also enhanced the angiogenesis ability of EA. hy926 cells. In addition, the expressions of intracellular VEGF and extracellular VEGFR (Flk-1) were regulated upward in a time-dependent manner. In conclusion, the proliferation of vascular endothelial cells and Vascula angiogenesis were improved under the condition of indirect co-culture.
Carcinoma, Hepatocellular ; pathology ; Cell Proliferation ; Coculture Techniques ; Culture Media, Conditioned ; Endothelial Cells ; cytology ; Hep G2 Cells ; Human Umbilical Vein Endothelial Cells ; Humans ; Liver Neoplasms ; pathology ; Neovascularization, Pathologic ; Tumor Microenvironment ; Vascular Endothelial Growth Factor A ; metabolism ; Vascular Endothelial Growth Factor Receptor-2 ; metabolism

Objective To understand the status of Toxoplasma gondii infection in the population of Pu er City so as to pro-vide the evidence for formulating the strategy of toxoplasmosis control. Methods The population from Jingdong Jinggu and Menglian counties in Pu er City was surveyed IgG of T. gondii in serum was detected by ELISA. Results Totally 906 resident serum samples were detected and the IgG positive rate was 24.2%. The positive rates were higher in the aged groups of 30-39 years and 60-69 years and the difference among different aged groups was significant χ2=17.77 P 0.01 . There were no si-gnificant differences between different sexualities and among different educational levels and living habits P 0.05 . The posi-tive rates were 26.6% 194/730 15.5% 22/142 and 8.8% 3/34 in farmers students and other occupations respectively and there was a significant difference among them χ2=12.51 P 0.01 . The positive rates were 23.3% 198/849 and 36.8%21/57 in the farmers who had the habit of rearing pigs in pens and the farmers who had the habit of free ranging pigs respec-tively and there was a significant difference between them χ2=5.33 P 0.05 . Conclusion The IgG positive rate of T. go-ndii is very high in Pu er City and therefore the health education for toxoplasmosis control should be strengthened.

Recombinant expression plasmid of pET-28a (+)-goIL-2 was constructed by inserting the goose IL-2 gene without the signal peptide sequence into the prokaryotic expression vector pET-28a (+), and transformed into the bacterial competent E. coli BL21 (DE3) cells for expression. After IPTG induction, an expected protein band with molecular weight of 15.0 kD was observed on SDS-PAGE gel, recognized by monoclonal antibody against goose IL-2 in western-blotting assay. In the pET-28a (+) expression system, much of the recombinant goose IL-2 (rgoIL-2) was found in inclusion bodies with a portion of soluble protein. The monomer and multimers of soluble goose interleukin 2 proteins were observed in native electrophoresis. The rgoIL-2 proteins were purified by Ni-NTA column under a native condition. The rgoIL-2 soluble protein monomer was isolated by a quick protein isolation and purification system of AKTA FPLC and identified by native PAGE. Bioactivity analysis showed that the rgoIL-2 monomer stimulated the proliferation of goose lymphocytes in vitro. This will establish a basis for further study about the biological function and clinical application of goose IL-2.
Animals ; Escherichia coli ; genetics ; metabolism ; Geese ; genetics ; Inclusion Bodies ; metabolism ; Interleukin-2 ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; isolation & purification ; Solubility

OBJECTIVE: To investigate the effect of cordyceps sinensis (CS) extract and losartan (Los) on the expression of Klotho (Kl), P53, P21, and apoptosis in renal tubular epithelial cell NRK-52E induced by angiotensin II (Ang II), and to elucidate its therapeutical mechanism in Ang II induced renal tubular epithelial cell apoptosis. METHODS: NRK-52E cells were incubated with CS with or without Ang II for 24 hours. Experimental groups were divided according to the increasing concentrations of CS:0 (serving as controls), 5, 10, 20, 40, and 80 mg/L. The optimal concentration of CS was selected and cells were divided into 5 groups: controls, Ang II (1*10(-8) mol/L), Ang II (1*10(-8) mol/L)+CS (40 mg/L), Ang II (1*10(-8) mol/L)+Los (1*10(-5) mol/L), and Ang II (1*10(-8) mol/L)+CS (40 mg/L)+Los (1*10(-5) mol/L). After 24 hours, cell proliferation was evaluated by MTT assay. The mRNA and protein expression of Kl, P53 and P21 were measured by RT-PCR. Activity of caspase-3 was evaluated by caspase-3 activity assay Kit. Cell apoptosis was determined by Annexin V-FITC/PI double staining and flow cytometry. RESULTS: Certain concentrations of CS promoted the proliferation of NRK-52E cells and increased cells proliferation inhibited by Ang II (P<0.01 or P<0.05 ). Ang II significantly down-regulated the mRNA and protein expression of Kl, and up-regulated the levels of P53 and P21. Caspase-3 activity and apoptotic rates were decreased, too (all P values<0.01). CS or/and Los significantly increased the expression of Kl mRNA and protein down-regulated by Ang II, decreased P53 mRNA and protein expression, P21 mRNA and protein expression,and inhibited caspase-3 activity and apoptotic rates(all P values<0.05). No cooperative effects were observed in the two drugs (P>0.05). CONCLUSION CS can increase the expression of Kl down-regulated by Ang II, decrease P53 and P21 expression and caspase-3 activity, and reduce Ang II induced NRK-52E cell apoptosis, which may be part of its mechanism of the protective effects on hypertensive renal damage.
Angiotensin II ; pharmacology ; Apoptosis ; drug effects ; Caspase 3 ; genetics ; metabolism ; Cells, Cultured ; Cordyceps ; chemistry ; Cyclin-Dependent Kinase Inhibitor p21 ; genetics ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Epithelial Cells ; cytology ; metabolism ; Glucuronidase ; genetics ; metabolism ; Humans ; Kidney Tubules ; cytology ; metabolism ; Losartan ; pharmacology ; RNA, Messenger ; genetics ; metabolism ; Tumor Suppressor Protein p53 ; genetics ; metabolism

OBJECTIVE:To investigate the effects of different diuretic regimens on related indicatora in hepatocellular carcino-ma patients after hepatic resection. METHODS:A total of 60 hepatocellular carcinoma patients undergoing hepatic resection were randomly divided into study group (30 cases) and control group (30 cases). After surgery,study group was given Spironolactone tablet 100 mg+Furosemide tablet 40 mg,qd,after breakfast;Control group was given Spironolactone tablet 100 mg orally,bid,af-ter breakfast and at 16:00+Furosemide tablet 20 mg,qd,after breakfast. Electrolyte,liver and renal function of 2 groups were re-checked every 3 days;diuretic regimen was adjusted according to body weight and volume of liquid intake and output. The effec-tiveness indexes of diuretic treatment were observed in 2 groups,such as the decrease of body weight,case number of effective di-uretic treatment,frequency of intravenous dieresis,frequency of hydroxyethl starch use,frequency of night arination,postoperative hospitalization stay,the time of ascites extinction;serum sodium,serum potassium,serum creatinine,blood urea nitrogen,serum al-bumin,ALT,AST,GGT,the occurrence of ADR were observed before and after treatment. RESULTS:The nocturnal micturition frequency of study group was significantly lower than that of control group,with statistical significance (P<0.05). There was no significant difference in the decrease of body weight,response rate of diuretic treatment,frequency of intravenous dieresis,frequen-cy of hydroxyethyl starch use,postoperative hospitalization stay or the time of ascites extinction between 2 groups(P>0.05). After treatment,the levels of serum sodium and serum albumin in 2 groups were decreased significantly,compared to before treatment;the levels of blood urea nitrogen,serum potassium and ALT,AST,GGT in 2 groups were significantly higher than before treat-ment,with statistical significance(P<0.05). There was no statistical significance in the levels of serum sodium,serum potassium, blood urea nitrogen,serum albumin or ALT,AST,GGT between 2 groups(P>0.05). There was no statistical significance in the incidence of ADR between 2 groups(P>0.05). CONCLUSIONS:Both two diuretic regimens can effectively avoid the occurrence of postoperative ascites formation and related complication in patients hepatocellular carcinoma after hepatic resection. The drug regi-mens of spironolactore 100 mg,qd+furosemide 40 mg,qd after breakfast can significantly reduce nocturnal micturition frequency and improve medication compliance.