Aim: To study the effects of Clostridium difficile toxin A on the cytotoxity and apoptosis of human hep-atoma cell line SMMC-7721. Methods: Clone inhibiting experiment and MTT calorimetric assay were used to assay SMMC-7721 proliferation. Morphological changes relevant to the apoptosis and the DNA damage were analyzed by transmission electron microscope and single cell gel assay. The number of apoptosis cells and the expression of Bcl-2 and p53 protein were detected by the flow cytometry. Results: 0. 018- 4. 690 mg/L toxin A significantly decreased the colony information of SMMC-7721 cells, and greatly inhibited SMMC-7721 proliferation in a time- and concentration-dependent manner. Morphological changes related to apoptosis were evident under transmission electron microscope and DNA damage was detected by single cell gel assay when SMMC-7721 cultured with 4. 690 mg/L toxin A for 48 h. Toxin A 0. 0734. 690 mg/L induced apoptosis in SMMC-7721 cells from 6. 8% to 41. 8%. In addition, Toxin A 0. 293-4. 690 mg/L significant decreased Bcl-2 protein expression and increased p53 protein expression in SMMC-7721. Conclusion: These results showed that clostridium difficile toxin A had a significantly cytotoxicity in human SMMC-7721 which was attributed to toxin A induced apoptosis.

Acquired Immunodeficiency Syndrome ; immunology ; Adolescent ; Chemokines ; immunology ; Child ; Child, Preschool ; Female ; Humans ; Male

Adenocarcinoma ; metabolism ; pathology ; Adenocarcinoma, Mucinous ; metabolism ; pathology ; Adenocarcinoma, Papillary ; metabolism ; pathology ; Carcinoma, Signet Ring Cell ; metabolism ; pathology ; Follow-Up Studies ; Gene Expression Regulation, Neoplastic ; Humans ; Lymphatic Metastasis ; Neoplasm Invasiveness ; Organic Cation Transport Proteins ; biosynthesis ; genetics ; Organic Cation Transporter 2 ; Osteopontin ; Prognosis ; RNA, Messenger ; biosynthesis ; genetics ; Sialoglycoproteins ; biosynthesis ; genetics ; Stomach Neoplasms ; metabolism ; pathology ; Survival Rate

OBJECTIVETo obtain the data of the genotypes and allele frequencies of 9 short tandem repeat (STR) foci (D18S1364, D12S391, D13S325, D6S1043, D2S1772, D11S2368, D22-GATA198B05, D8S1132, and D7S3048) in Chinese subjects of Li and Han nationalities in Hainan Province.

METHODSA total of 154 randomly selected unrelated subjects of Li nationality and 112 unrelated local Han subjects from Hainan Province, along with 125 Han subjects recent immigrated to Hainan were enrolled in this investigation. Venous blood samples were obtained from the subjects to determine the allele frequencies at the 9 STR loci using multiplex primer extension assay. Statistical analysis was performed to determine the observed heterozygosities (Hobs), polymorphism information content (PIC), individual identification probability (Dp), accumulated power of discrimination, and cumulative rate of exclusion. The genetic polymorphisms of the 9 STR loci was analyzed to acquire the population genetics data.

RESULTSThe genotype and frequency distributions of the 9 STR loci were consistent with Hardy-Weinberg equilibrium. The Hobs, PIC, Dp, cumulative rate of individual identification (CDP), and cumulative chance of exclusion (CCE) showed no significant difference between the 9 foci.

CONCLUSIOND18S1364, D12S391, D13S325, D6S1043, D2S1772, D11S2368, D22-GATA198B05, D8S1132, and D7S3048 STR loci in Chinese Li and Han population in Hainan Province show a high probability of personal identification without obvious difference between the populations, suggesting their value in forensic science and population genetics.

Adolescent ; Adult ; Alleles ; Asian Continental Ancestry Group ; genetics ; China ; ethnology ; Female ; Gene Frequency ; Genetics, Population ; Genotype ; Humans ; Male ; Microsatellite Repeats ; genetics ; Middle Aged ; Polymorphism, Genetic ; Young Adult

OBJECTIVE To investigate the nuclear proteomes in mitochondrial DNA (mtDNA)-depleted A549 cells (Rho0 cells) and their parental cells (Rho+ cells),and to learn more about the nuclear responses to mitochondrial dysfunction.METHODS The nuclear proteomes of Rho and Rho + cells were characterized by two dimensional electrophoresis (2-DE) and SELDI-TOF ProteinChip technologies,the differentially expressed protein- spots were identified by MALDI-TOF mass spectrum (MS),the nucleophosmin and P53 expression were detected by Western blotting assay,and the mitochondrial memhrane potential (MMP) was measured by the laser scanning confocal microscope.RESULTS 2-DE results showed 11 protein-spots were down-regulated and 21 protein-spots were up-regulated in Rho0 cell nuclei.SELDI-TOF MS analysis with NP20 ProteinChips revealed 4 protein-peaks decreased in Rho0 cell nuclei.One down-regulated protein-spot was identified as elF-6,and 4 up-regulated proteinspots were identified as nucleophosmin,SFRS1,SFRS3 and hnRNP G,respectively.The increased expression of nucleophosmin in Rho0 cells was verified by Western blotting.Based on the clues from proteomic analysis,P53 expression in Rho0 cells was higher than in Rho + cells,and MMP was consistent in Rho + and Rho0 cells.CONCLUSION mtDNA-depletion induces nuclear proteome alteration.Rho0 cells can be used as a model to study the crosstalk between mitochondrion and nucleus.

Objective To investigate the pathogenesis of peripheral blood mononuclear cells(PBMC) nuclear factor kappa-?B(NF-?B) in children with primary nephritic syndrome(PNS) and the effect of astragalus on the activity of NF-?B.Methods Twenty-five children with PNS and 20 normal children were studied.Isolated PBMC were separated from 5 mL venous blood in asepsis condition.NF-?B stimulator,NF-?B inhabitor and astragalus were added into the different tubes of PBMC,respectively.The nuclear protein was extracted from the pellets and the optical density(A) values of nuclear protein was measured by enzyme-linked immunosorbent assay(ELISA).Results The activity of PBMC NF-?B in PNS group was higher than that in normal group(P0.05).Astragalus could decrease the activity of PBMC NF-?B which had been stimulated by interleukin-1?(IL-1?)(P

Objective: To investigate the changes of adhesion molecules in peripheral blood in patients with recurrent cerebral infarction. Methods: By cytometry and enzyme-linked immunosorbent assay, the expression of integrin Mac-1 ?-sub-unit(CD18) ,intercellular adhesion molecule-3(CD50) intercellular adhesion molecule-1 (CD54) ,very late antigen-4 ?-subunit and p-subunit(CD49d and CD29),CD44 and L-selectin(CD62L) in lymphocyte,the expression of platelet membrane glycopro-tein I b- I a complex a-subunit (CD41), P-selectin (CI)62p) ,the serum level of soluble P-selectin(sP-selectin) and soluble intercellular adhesion molecule-1 (sICAM-1) were measured in 33 recurrent cerebral infarction patients and 44 patients with previous ever-single cerebral infarction history. Results: Compared with previous ever-single cerebral infarction history patients, the positive percentage of CD18,CD50,CD54 ,CD49d,CD29,CD44 and CD62L in lymphocytes ,CD41 on platelet did not significantly change,while the positive percentage of CD62p on platelet and the level sP-selectin,sICAM-1 were significantly higher (P

Objective To study the effects of rehabilitation(RT)on cardiac function in cerebral infarct (CIF)patients with cardiac insufficiency(CIS).Methods Fifty-nine CIF patients with CIS were randomly divid- ed into a treatment group(T group,n=29)and a control group(n=30),and all patients were treated with routine pharmacotherapy for 2 months.In addition,RT was administrated in the T group at the same time.The grading of the New York Heart Association(NYHA)and the changes in cardiac function associated index were observed in both groups before and after treatment.Results Compared with the control group,NYHA grades,left ventricle ejection fraction(LVEF),the levels of brain natriuretic peptide(BNP)in the blood plasma,and the 6min walking range of the T group patients were all significantly improved after treatment(P＜0.05).Conclusion RT can improve car- diac function in CIF patients with CIS.

This study is to investigate the effect and mechanism of puerarin on DNA damage of HaCaT cells induced by UVB. Puerarin pre-treated cells were irradiated with UVB at 30 mJ x cm(-2). Twenty four hours after irradiation, DNA damage was detected by comet assay, ceramide was measured by thin layer chromatography and gas chromatography, intracellular free calcium ion was analyzed by flow cytometry, the phosphorylation level of p38 protein was examined by Western blotting method. Levels of DNA damage, ceramide, free calcium ion and p-p38 protein were elevated in UVB model cells. Contrary to the model group, all indicators above were reduced in all groups pre-treated by puerarin. Puerarin restrains the ceramide accumulation to block downstream p38 MAPK pathway and calcium ion rising, therefore reduces DNA damage in HaCaT cells induced by UVB.
Calcium ; metabolism ; Cell Line ; Ceramides ; metabolism ; DNA Damage ; drug effects ; radiation effects ; Down-Regulation ; Humans ; Isoflavones ; pharmacology ; Keratinocytes ; cytology ; metabolism ; Phosphorylation ; Signal Transduction ; drug effects ; Ultraviolet Rays ; adverse effects ; p38 Mitogen-Activated Protein Kinases ; metabolism

Endotoxemia ; metabolism ; pathology ; HMGB1 Protein ; Humans ; Liver ; metabolism ; pathology