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

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 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.

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.

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

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

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

Background and purpose:Ionizing radiation(IR) activates the early growth response- I(Egr1) promoter through specific cis-acting sequences termed CArG elements by production of radical oxygen intermediates(ROls).Egr-EG,an expression vector pCIneo containing CArG elements cloned upstream of the cDNA for human recombinant GM-CSF,was used to treat hematopoietic damage due to chemotherapy.Commonly used chemotherapeutic agents can cause tumor cell death by producing DNA damage and generating ROIs.We therefore hypothesized that clinically employed chemotherapeutic agents that increase ROIs could also be employed to activate Egr-EG in a chemoinducible gene therapy strategy.This study was done to explore the chemo-protective effect of the expression of hematopoietic growth factors regulated by Egr-1 promoter on chemotherapy induced damage. Methods:The human GM-CSF cDNA and EGFP cDNA were linked together with internal ribosome entry site(IRES) and then inserted into the eukaryotic expression vector pCI-neo with the Egr-1 promoter(Egr-EG),and was further transduced into human bone marrow stromai cell lines HFCL(HFCL/EG).The HFCL/EG cells were transplanted i.v.into BI6 melanoma in C.B-17 combined immunodeficient(SCID) mice.5-FU was given i.p.on day 3 and 4.The white blood cell amount in peripheral blood,the expression of EGFP and GM-CSF in human stromal cell engrafted in recipient mice were detected by flow cytometry,RT-PCR,Western blot and colony-forming units for granulocytes and macrophages(CFU-GM),respectively.Results:In contrast to the two control groups,HFCL/EG(the Egr-regulatory element-derived expression of GM-CSF gene therapy) resulted in a proportional increase in the number of the white blood cell after chemotherapy,no significant diifferences were found for CFU-GM in bone marrow cells and the inhibition ratio on tumor in recipient mice.Chemotherapy could markedly increase the expression of EGFP and GM- CSF mRNA/protein as compared with that of non-chemotherapy control groups and HFCL group.Conclusion: Chemoinducible GM-CSF gene therapy regulated by Egr-1 promoter can ameliorate the toxic effect on 5-FU chemotherapy-inducible hematopoietic damage.

Objective To investigate a feasibility of treatment planning in thoracic esophageal carcinoma with 3-deoxy-3-fluorothymidine (FLT) PET-CT and to compare with fluorodeoxyglucose (FDG) PET-CT based on dosimetric analysis.MethodsTwenty-two patients with esophageal squamous cell carcinoma detected by FLT and FDG PET-CT were enrolled.The gross tumor volumes ( GTV ),clinical target volume(CTV) and planning target volume ( PTV ) were delineated using treatment planning system of Philips Pinnacle3 based on the optimal threshold of FLT and FDG PET-CT respectively,and to make two groups simulation treatment planning.The parameters of dose-volume histograms in two groups planning were compared in the similar direction and ensuring prescribed dose line surround 95% target volume.Results The values of GTV,CTV and PTV in FLT PET-CT planning were less than those of FDG,that dose received by spinal cord in two planning were not significantly yet ( t = - 1.60,- 1.55,all P ＞ 0.05 ).While,the values in mean lung dose,V5,V10,V30,V40 and V50 of bilateral lung,mean heart dose,and V30 of heart in FLT PET-CT planning were significant lower than those of FDG( t = -5.442 - -2.637,all P ＜0.05).Conclusions Compared with FDG,FLT PET-CT based treatment planning brings potential benefits for lungs and heart.