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At least 919 records · Page 51Linked to original sources

PubMed2003

[VEGF production and its receptor expression of MPhis in peritoneal fluid].

AIM: To explore the role of microenvironment of peritoneal fluid in pathogenesis of endometriosis, the source and its regulation in peritoneal fluid. METHODS: Peritoneal Mphis from 14 patients with endometriosis (endometriosis group)and 10 normal women ( control group) were cultured in vitro. Either 17-l estradiol, progesterone and LPS or their combinant were added to cultured peritoneal Mos from control group. VEGF levels in culture supernatant of M(Ds from two groups were detected by ELISA. Culture supernatant of serum-free media in two groups were added to the cultured endothelial cells. Effect of the supernatant on endothelial cell proliferation activity was examined by MTT colorimetry at wavelength of 450 nm. The expressions of Fit-1 and Flt-4 on the Mos from two groups were determined by immunohistochemical staining. RESULTS: The VEGF level in culture supernatant of Mps from endometriosis group was significantly higher than that from control group (P < 0. 05 ), and had no cyclic variation (P > 0. 05). As compared with LPS, estrogen and progentrogen could elevate notably the level ofVEGF secreted by Ms ( P < 0. 05 ). However, the regulation of MWs secreted VEGF level by estrogen and progestoron had no marked difference. The Mos activated by LPS could strengthen the regulation of VEGF secreted by Mphis under estrogen and progestron stimulation. Culture supernatant of serum-free media for Mos could promote significantly the proliferation of endothelial cells ( P < 0. 05 ). Fit-land Flt-4 on Mos was expressed without cyclic variation (P>0.05). CONCLUSION: The VEGF may be secreted by Mos in peritoneal fluid of endometriosis group. The endothelial cell proliferation and angiogenesis are enhanced by change of microenvironment of peritoneal fluid.

Adult↗

PubMed2003

[Effect of glia cell line-derived neurotrophic factor on cultured spinal motor neurons from embryonic rat].

AIM: To observe the effects of various concentrations of gli-a cell line-derived neurotrophic factor ( GDNF) on the growth of cultured spinal motor neurons from embryonic rat. METHODS: Neurons were isolated from embryonic rat spi-nal ventral tissue and then were primarily cultured in vitro. Morphological feature of neurons was examined by immuno-cytochemical staining with mAb SMI-32. The effect of GDNFon growth of spinal motor neurons was detected by MTT colorimetry. RESULTS: GDNF could significantly enhance the survival and neurite outgrowth of rat spinal motor neurons cultured in vitro in dose-dependent tendency. CONCLUSION: Varying concentrations of GDNF can enhance the growth of embryonic rat spinal motor neurons to various extent.

Animals↗

PubMed2003

[Effect of antisense CD40 RNA on CD40 expression and proliferation of EB virus-transformed B lymphocytes].

AIM: To investigate the changes of CD40 expression and proliferation pattern of EB virus-transformed B lymphocytes that had been transfected by antisense CD40 RNA. METHODS: Eukaryotic expression vector pcDNA3/CD40 of human CD40antisense RNA was constructed by means of T-A cloning and subcloning, and then was transfected into EB virus-transformed B lymphocytes. Changes of CD40 expression on the B cells and their proliferation were tested by flow cytometry and MTT colorimetry assay, respectively. RESULTS: CD40 expression and proliferation of the B cells transfected with antisense CD40 RNA were significantly lower than those of the cells transfected with pcDNA3 alone (P < 0. 01). CONCLUSION: Antisense CD40 RNA may be served as an effective means of regulating immune function, and the CD40 gene itself also play an important role in cell proliferation.

B-Lymphocytes↗

PubMed2003

[The influence of the extract of lumbricus on the production of NO and TNF-alpha by mouse MPhi and splenocytes].

AIM: To explore the effect of the abstracts of lumbricus on the secretion of NO and TNF-alpha by mouse Mphi s and splenocytes. METHODS: Murine Mphi s and spleen cell were co-cultured with various doses of lumbricus abstracts for 24 hours and then the supernate was collected. The levels of NO and TNF-alpha were detected by diazotization reaction and MTT colorimetry, respectively. RESULTS: Compared with control group, 0.1 g/L of lumbricus abstracts could increase the NO level and antagonize the inhibition of dexamethasone(Dex). 1 x 10(-4), 1 x 10(-3) g/L of lumbricus abstracts could increase TNF-alpha level and also antagonize the inhibition of Dex on the secretion of TNF-alpha by Mphi s and splenic cells. CONCLUSION: The abstracts of lumbricus can activate Mphi s and splenic cells to secrete NO and TNF-alpha and antagonize the inhibition effect of Dex on these cells.

Animals↗

PubMed2004

[Apoptosis and modulatory mechanism of human AHH-1 T lymphocytes induced by gamma-irradiation].

AIM: To explore the apoptotic characteristics and modulatory mechanism of human AHH-1 T lymphoblast induced by ionizing radiation and provide an experimental basis for preventing and treating immune damage in acute radiation sickness. METHODS: TdT-mediated dUTP nick end labeling (TUNEL) and May-Grunwald Giemsa (MGG) staining, MTT colorimetry and immunohistochemical staining were used to detect the T lymphocyte apoptosis, cell survival and the expressions of Bax, Bcl-XL and caspase-3 proteins. RESULTS: (1) Over a definite dose range, apoptotic rate of AHH-1T lymphocytes increased with the elevation of radiation doses, while the lymphocytic survival rate decreased sharply, both showing a good dose-effect relationship. (2) The expression of apoptosis-accelerating protein Bax enhanced obviously with the increase of radiation doses, while apoptosis-inhibiting protein Bcl-XL trended to persistent decline. (3)The activity of activated apoptosis-executing protein caspase-3 heightened significantly with increased radiation doses, which revealed a better corresponding relationship to apoptotic rate. CONCLUSION: A great number of apoptotic AHH-1 T cells may be one of major accounts for the lymphocyte rapid reduction and depressed organism immune function induced by irradiation. Bax, Bcl-XL and caspase-3 proteins play an important role in the regulation of radiation-induced T cell apoptosis.

Apoptosis↗

PubMed2003

[Influence of bondage stress on Th1/Th2 cytokines and growth of tumors in mice bearing S180 tumor cells].

AIM: To observe the influence of bondage stress on Th1/Th2 cytokines and growth of tumor in mice bearing S180 tumor cells. METHODS: 2 x 10(6) S180 tumor cells were injected subcutaneously into the murine right oxter,and bondage stress was imposed on mice for 8 hours per day,ten days running. Then T lymphocyte proliferation was detected by MTT colorimetry. IFN-gamma and IL-2 secreted by splenocytes was measured by mitogen-activating lymphoblast assay and macrophage NO production assay. At the same time, the levels of IL-4 and IL-10 in serum were detected by ELISA, and the weight of the tumors and thymuses were measured. RESULTS: Compared with mice only injected with S180 tumor cells, tumor bearing mice suffered from bondage stress showed decreased T lymphocyte proliferation, thymus index, and levels of IFN-gamma and IL-2, but increased levels of IL-4 and IL-10(P<0.05), and tumor weight (P<0.01). CONCLUSION: Bondage stress could aggravate the suppression of cellular immunity in mice bearing S180 tumor and promote the T cells to shift towards Th2 type cells, which may be one of the mechanisms of promoting tumor growth by bondage stress.

Animals↗

PubMed2003

[The effect of CTLA4Ig-modified dendritic cells on proliferation and cytotoxicity of lymphocytes in-vitro].

AIM: To study the effect of dendritic cells (DCs) modified by CTLA4Ig on T-cell proliferation and the cytotoxic T lymphocyte killer activity. METHODS: The expressing vector pG/CTLA4Ig was transfected into DCs via lipofectamine reagent mediation. CTLA4Ig fusion protein secreted by DCs in the culture supernatant was confirmed by sandwich ELISA and SDS-PAGE. Peripheral blood mononuclear cells (PBMC) from C57BL/6 mice (as reaction cells) and DCs modified or unmodified by CTLA4Ig (as stimulation cells) were co-cultured for 6 days. MTT colorimetry was used to detect lymphocyte proliferation. Lactate dehydrogenase release method and sandwich ELISA assay was used to examine the cytotoxic activity and T-cell apoptosis. RESULTS: CTLA4Ig fusion protein and DCs modified by CTLA4Ig could significantly inhibit lymphocyte proliferation response and cytotoxic activity of specific cytotoxic T lymphocytes, and induce T-cell apoptosis, while unmodified-DCs could markedly induce lymphocyte proliferation response. CONCLUSION: The DCs of stably expressing CTLA4Ig fusion protein could not only markedly inhibit T-cell proliferation and cytotoxic activity of CTL, but also induce T-cell apoptosis.

Abatacept↗

PubMed2003

[Effect of resveratrol alone and its combination with cyclosporin A on the immune function of human peripheral blood T lymphocytes].

AIM: To investigate the effect of resveratrol (Res) alone and its combination with cyclosporin A (CsA) on the proliferation of human peripheral blood T lymphocytes (hPBTCs), transformation into lymphoblasts, as well as IL-2 and INF-gamma production. METHODS: The proliferation of hPBTCs was detected by MTT colorimetry. The lymphocyte transformation rate was examined by morphologic method. The levels of IL-2 and INF-gamma in lymphocyte culture supernatant were detected by ELISA. RESULTS: When the concentration of resveratrol was over 2.5 mg/L, especially 10 mg/L, the proliferation and transformation rate of hPBTCs decreased significantly (P<0.05). The levels of IL-2 and INF-gamma in lymphocyte culture supernatant reduced markedly (P<0.05), on condition that the concentration of resveratrol was over 5 mg/L. The combination of more than 2.5 mg/L of resveratrol with cyclosporine A had synergism. CONCLUSION: Resveratrol can suppress notably the proliferation and transformation of human lymphocytes and the combination of resveratrol at a given concentration with cyclosporine A can enhance immune suppression.

Cyclosporine↗

PubMed2003

[Cooperative effect of rhIL-2 and adriamycin-magnetic albumin microsphere targeting therapy on tumor].

AIM: To observe the synergistic inhibitory effects of rhIL-2 and adriamycin magnetic albumin microsphere(ADM-MAM) targeting therapy on tumor and to explore their antitumor mechanism. METHODS: The antitumor activity was observed using the tumor weight as index. The killer activity of natural killer (NK) cells and the lymphocyte transformation were examined by the LDH release assay and MTT colorimetry, respectively. The apoptosis of tumor cells and the expressions of p53,Fas and FasL were examined by flow cytometry. The expressions of IL-2 and IL-12 were determined by RT-PCR. RESULTS: Compared with the control group, in ADM-MAM targeting therapy group and combination therapy (ADM-MAM and rhIL-2 therapy) group, tumor weight, killer activity of NK cells, and the lymphocyte transformation;were obviously decreased; while the expression of Fas, FasL, IL-2 and IL-12 were markedly increased. CONCLUSION: ADM-MAM targeting therapy exerts anti-tumor effects with fewer side effects, which can be enhanced when combined with rhIL-2 therapy. Such synergistic anti-tumor effects may be realized by stimulating the proliferation of T cells and growth of NK cell to enhance cellular immunological function.

Albumins↗

PubMed2003

[Construction, expression and functional characterization of disulfide-stabilized anti-hepatocarcinoma single chain Fv fused with truncated Pseudomonas exotoxin].

AIM: To prepare fusion protein of disulfide-stabilized anti-hepatocarcinoma single chain Fv(dsFv)and truncated Pseudomonas exotoxin(PE38). METHODS: The gene encoding the fusion protein was cloned into the prokaryotic expression vector pTIG which was then transformed into E.coli origami(DE3). The expressed product was purified through Ni-NTA agarose affinity chromatography column. The cytotoxicity of the purified dsFv-PE38 was detected by MTT colorimetry. RESULTS: The immunotoxin dsFv PE38 was expressed in E.coli origami(DE3) in a soluble form, which accounted for nearly 21% of the total bacterial soluble proteins. The results of cytotoxic assay showed that the dsFv-PE38 could specifically kill hepatocarcinoma cells, while spared the normal hepatic cells. CONCLUSION: The immunotoxin dsFv-PE38 may have some potential value in the treatment of hepatocarcinoma.

ADP Ribose Transferases↗

PubMed2003

[Immunogenicity of DNA vaccine encoding fusion protein of Mycobacterium tuberculosis Ag85B and MPT64].

AIM: To study the immunogenicity of a DNA vaccine encoding fusion protein of Mycobacterium tuberculosis Ag85B and MPT64. METHODS: C57BL/6 mice were intramuscularly injected respectively with PBS(A group), plasmid vector (B group), pcDNA/Ag85B(C group), pcDNA/MPT64(D group), and pcDNA/Ag85B MPT64 (E group) for three times with 2 week intervals. The mice were sacrificed 4 weeks after the final immunization. The total specific antibody levels were detected by ELISA and simultaneously spleen lymphocyte proliferation response to PPD and IFN-gamma level in culture supernatant of the PPD stimulated spleen lymphocytes was measured by MTT colorimetry and ELISA, respectively. RESULTS: The Ag85B, MPT64 and fusion gene Ag85B MPT64 DNA vaccines could induce anti-PPD antibody production as well as antigen specific lymphocyte proliferation and high levels of IFN-gamma in immunized mice. CONCLUSION: The Ag85B MPT64 fusion gene can induce specific cellular immunity and humoral immunity in immunized mice.

Acyltransferases↗

PubMed2004

[Antibody production and its neutralization of Abeta42's cytoxicity in BALB/c mice induced by Abeta42 and its subunit vaccines].

AIM: To explore the production of anti-Abeta42 antibody after immunization with Abeta42 and its subunit peptide vaccines. METHODS: Seventy five male BALB/c with the age of 6 weeks were randomly divided into 5 groups, namely, control group, Abeta42 group, Abeta(36-42) group, Abeta(1-15)group and F Abeta(1-15) group. The BALB/c mice were immunized four times with PBS+MF59 adjuvant, Abeta42+MF59, Abeta(36-42)+heptalysine (MAP)+MF59, Abeta(1-15)+MAP+MF59 and Abeta(1-15)+MAP+Freud's adjuvant, respectively. The titers of specific antibodies in sera and supernatants of brain tissue homogenates from the immunized mice in every group were detected by indirect ELISA. After Abeta42, Abeta(36-42), Abeta(1-15) and F Abeta(1-15) were co-cultured together with cultured PC12 cells for 7 days, the cytotoxicity of the 3 antigen peptides to PC12 cells were determined by MTT colorimetry. In addition, after immune sera from each group were added to culture medium containing 20 mg/L Abeta42 and co-cultured with PC12 cells for 7 days, the survival rate of PC12 cells were examined by MTT assay. RESULTS: The production of anti-Abeta42 antibodies was detected in sera of each experimental group after the second time immunization, and the titer of antibody rose with the increase of immunizing times. In addition, the anti-Abeta42 antibody with low titer was also detected in supernatants of brain tissue homogenates. The Abeta42 could reduce the survival rate of PC12 cells, whereas Abeta(36-42) and Abeta(1-15) had no obvious effect on survival rate of PC12 cells. After immune sera from 4 experimental groups and Abeta42 were co-cultured with PC12 cells, their survival rate was found improved. CONCLUSION: Combination of Abeta42 and its subunit (Abeta(36-42) and Abeta(1-15)) vaccines with MF59 adjuvant can induce BALB/c mice to produce anti-Abeta42 antibody. The antibody may neutralize the cytotoxicity of Abeta42.

Adjuvants, Immunologic↗

PubMed2004

[The effect of polyporus umbellatus polysaccharide on the immunosuppression property of culture supernatant of S180 cells].

AIM: To investigate the effect of polyporus umbellatus polysaccharide (PUPS) on the immunosuppression property of tumor cell line S180 culture supernatant. METHODS: The inhibitory effects of the culture supernatant of S180 cells in the presence or absence of PUPS on the ConA-induced mouse splenocyte proliferation, IL-2 production, killer activity and the reactivity of CTLL-2 cells to IL-2 were detected by MTT colorimetry. The effect of the culture supernatant on the IL-2Ralpha expression on murine splenocytes was detected by flow cytometry. RESULTS: The culture supernatant of S180 cells could strongly inhibit immunity in terms of the above five indexes, while PUPS could reverse the immunosuppression. CONCLUSION: PUPS can offset the immunosuppression of the supernatant from S180 cell culture, which may be mediated by down-regulating the synthesis and/or secretion of immunosuppressive substance by S180 cells.

Animals↗

PubMed2004

[Experimental study on immune responses induced in mice by gp120 gene vaccine of Chinese HIV-1 strain].

AIM: To construct the gp120 DNA vaccine of Chinese HIV-1 strain and evaluate the immune responses induced with it in BALB/c mice. METHODS: The recombinant expression vector pVAX1-GP120 was constructed by inserting HIV gp120 gene into the eukaryotic expression vector pVAX1 and confirmed with EcoR I/Pst I and DNA sequencing. BALB/c mice were immunized with pVAX1-GP120 and pVAX1 respectively. The levels of serum anti-HIV antibody and IFN-gamma of the immunized mice were detected by ELISA. The proliferation of splenocytes was determined by MTT colorimetry and the specific cytotoxic T lymphocytes (CTLs) response by LDH assay. RESULTS: Restriction enzymes digestion analysis and DNA sequencing results revealed that the pVAX1-GP120 had been constructed successfully. The titer of anti-HIV antibody and the IFN-gamma level in mice immunized with the pVAX1-GP120 were higher than those in mice immunized with pVAX1 respectively (P<0.01). As compared with mice immunized with pVAX1 alone, the cytotoxic activity of specific CTLs and antigen-specific lymphoproliferative responses in mice immunized with pVAX1-GP120 were significantly enhanced (P<0.01). CONCLUSION: Specific cellular and humoral immune responses in mice can be induced with gp120 gene vaccine of Chinese HIV-1 strain, which lays the foundation for further development of therapeutic HIV vaccine against HIV-1 infection.

AIDS Vaccines↗

PubMed2004

[Effects of TGF beta 1 on the cell cycle of irradiated human lung fibroblasts and its signal transduction mechanism].

AIM: To investigate the effects of TGF beta 1 on the proliferative activity and cell cycle of normal and gamma ray-irradiated human lung fibroblasts (HLFs). METHODS: HLFs were cultured and irradiated with 3Gy gamma rays and then stimulated by TGF beta 1 at 0.01, 0.1, 1 and 10 microg/L. The proliferative activity and cell cycle were detected by MTT colorimetry and FACS. Furthermore, the expressions of signal proteins Smad3/4 in HLFs were observed by immunohistochemical staining and FACS. RESULTS: TGF beta 1 of 10 microg/L could promote the proliferation of HLFs, with the number of cells in S phase increased. It also could enhance the expression of Smad3 protein and promote the nuclear translocation of Smad4 protein. CONCLUSION: A given dose of TGF beta 1 can promote the proliferation of gamma rays-irradiated HLFs by regulating their cell cycle in which the activation of Smad pathway may play a role.

Cell Cycle↗

PubMed2004

[Role of Akt/NF-kappa B signal transduction pathway in murine glomerular mesangial cell proliferation induced by immune complex].

AIM: To explore whether immune complex (IC) can directly induce glomerular mesangial cells(MCs) proliferation and the role of Akt/NF-kappa B signal pathway in the proliferation. METHODS: The mice were divided into control, stimulation and oligodeoxynucleotide(ODN) groups. In ODN group, MCs isolated from mice were transfected with Akt1 sense, mismatched or antisense ODN for 8 h, respectively, by using lipofectin, control and stimulation groups were incubated with lipofectin for 8 h. Then stimulation and ODN groups were incubated with aggregated IgG(AIgG)(a standard IC model), while the control group with monomeric IgG. MTT colorimetry was used to detect MCs proliferation. Distribution of MCs in cell cycle was analyzed by flow cytometry. Cyclin D1 mRNA and its protein expression were determined by RT-PCR and Western blot, respectively.The activity of NF-kappa B in MCs was determined by EMSA. RESULTS: AIgG activated NF-kappa B, upregulated cyclin D1 mRNA and its protein expression, and induced majority of MCs to enter S-phase in cell cycle. Akt1 antisense ODN specifically decreased AIgG-induced NF-kappa B activation, cyclin D1 mRNA and its protein expression, and then inhibited MCs to progress to S-phase and cell proliferation. Sense ODN and mismatched ODN had no such effects. CONCLUSION: IC can directly stimulate MCs proliferation through Akt/NF-kappa B signal pathway, suggesting that NF-kappa B probably be a useful molecule for targeted therapy in IC-mediated MC overproliferation.

Animals↗

PubMed2004

[Expression and activation of MAPK pathway signaling molecules in human breast cancer cell lines].

AIM: To detect the expression and status of extracellular regulatory kinase 1 and 2 (ERK1/2) and its upstream kinase MEK1/2 proteins in four breast cancer cell lines MCF-7, Bcap-37, SK-BR-3 and T47D and study the effects of cyclophosphamide and epirubicin on the growth of the cell lines and on the expression and status of the signaling molecules. METHODS: Western blot was used to examine the expression and status of MEK1/2 and ERK1/2 proteins in these cells and the effects of these two drugs on them. The effects of the two drugs on the proliferation of these breast cancer cell lines were detected by MTT colorimetry. RESULTS: The levels of expression and phosphorylation of MEK1/2 and ERK1/2 proteins in four breast cancer cell lines increased notably as compared with those in MCF-10 cells. Both drugs could inhibit the proliferation of breast cancer cells. And the levels of expression and phosphorylation of MEK1/2 and ERK1/2 proteins in breast cancer cell lines treated with the drugs were markedly lower than those in untreated breast cancer cells. CONCLUSION: Overexpression and phosphorylation of MEK and ERK may play an important role in the generation and development of human breast cancer. The inhibitory effect of cyclophosphamide and epirubicin on proliferation of the breast cancer cells may be by means of inhibiting expression and phosphorylation of MEK and ERK.

Antibiotics, Antineoplastic↗

PubMed2004

[Effects of insulin-like growth factor II on regulating nitric oxide synthase gene expression in mouse osteoblast-like MC3T3-E1 cells].

OBJECTIVE: To study the effects of insulin-like growth factor II (IGF-II) on regulating the levels of nitric oxide (NO) and the mRNA transcriptions of inducible nitric oxide synthase (iNOS) and endothelial NOS (eNOS) in mouse osteoblast-like cells. METHODS: Mouse osteoblastic cell line MC3T3-E1 was selected as the effective cell of IGF-II. After the cells were treated with IGF-II at different concentrations for different intervals of time, MTT colorimetry was used for examining the cell proliferation. Nitrate reductase method was applied for detecting the NO concentrations in cell culture supernatants and RT-PCR employed for determining the levels of cellular iNOS and eNOS mRNAs. RESULTS: After the MC3T3-E1 cells were treated with IGF-II at the dosages of 1 microg/L for 72 h, 10 and 100 microg/L for 24, 48 and 72 h respectively, all the MTT values increased markedly (P < 0.05 or P < 0.01). After the cells were treated for 48 and 72 h at the dosage of 100 microg/L IGF-II respectively, the levels of NO in the supernatants of cell cultures and cellular iNOS mRNA decreased significantly (P < 0.01). However, the levels of eNOS mRNA in the cells treated with any of the IGF-II dosages for the different times were stable (P > 0.05). CONCLUSIONS: IGF-II at the dosages of 1 approximately 100 microg/L showed the effects on promoting proliferation, which as probably due to the maintenance of low NO levels. Inducible NOS gene expression at the level of transcription was down regulated in the MC3T3-E1 cell treated with higher dosage of IGF-II (100 microg/L) but eNOS mRNA was not, which might be one of the mechanisms for the maintenance of low NO levels.

3T3 Cells↗