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Objective: To investigate the effects of medicated rat serum containing Gengnianchun (GNC) decoction and its protection to pheochromocytoma cells (PC12 cells) from amyloid beta (Abeta)(25-35)-insulted apoptosis and to find the possible mechanism. Methods: Medicated rat serum was prepared by administering ovariectomized Sprague-Dawley (SD) rats with GNC decoction. The effects of medicated rat serum on viability of PC12 cells were evaluated by cell counting kit-8 (CCK-8) assay. The PC12 cells were cultured with different doses of Abeta(25-35) to induce a model of Alzheimer's disease in vitro. Then, the protective effects of medicated rat serum on Abeta(25-35)-insulted PC12 cells were evaluated by using CCK-8 assay to detect the cell viability, using Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) flow cytometry to detect cell apoptosis rate and using Western blotting assay to analyze the expressions of Bcl-2, Bax and active caspase-3 proteins. Results: PC12 cells cultured with 20% medicated rat serum containing GNC decoction for 24 h or 48 h had higher viability than those cultured with normal culture medium (P<0.05). After 24- or 48-hour treatment of different concentrations of Abeta(25-35), cell viabilities were all decreased as compared with normal medium (P<0.05). Cells underwent apoptosis, which showed the neurotoxicity of Abeta(25-35). The cell apoptosis induced by Abeta 25-35 was significantly decreased in PC12 cells which were pretreated with 20% medicated rat serum or nerve growth factor (NGF) according to CCK-8 assay and Annexin V-FITC/PI flow cytometry (P<0.05). The ratio of Bax expression to Bcl-2 expression and the expression of active caspase-3 were decreased in the cells treated with medicated serum or NGF as compared with the cells cultured with Abeta(25-35) only. Conclusion: The GNC-medicated rat serum at concentration of 20% can promote viability of Abeta(25-35)-insulted PC12 cells and decrease the cell apoptosis by regulating the expressions of Bcl-2, Bax and active caspase 3.
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Postmenopausal osteoporosis can be classified as an inflammatory, or even an autoimmune condition, hence new therapeutic " immune" targets. Some recently published works in this area are summarized, which may help to find a noval strategy for the treatment and prevention of postmenopausal osteoporosis.
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Objective:To investigate the expression of a new cytokine,thymic stromal lymphopoietin (TSLP) and its receptor TSLPR in the villi of human first trimester gestation.Methods:Villi were collected from women who had undergone an artificial abortion at 7-11 weeks of normal gestation.The trophoblast cells (Tros) were isolated and cultured.The total RNA was extracted using TRIzol reagent,from both villi and the Percoll-gradient-isolated Tros,then the DNA fragments of hTSLP and hTSLPR were amplificated by RT-PCR.Villous tissues were detected for TSLP by immunohistochemistry (IHC) and Western blot.Immunocytochemistry (ICC) was carried out on cultured trophoblast cells for TSLP/TSLPR expression.Levels of TSLP in the supernatants were detected by ELISA.Results:Normal villi and the cultured Tros transcript were found to express TSLP/TSLPR mRNA and secreted TSLP protein.In addition,TSLP receptor was also expressed on trophoblasts.Conclusion:Both TSLP and its receptor are expressed on villi and trophoblast cells,which suggests that TSLP plays an important role in maternal-fetal immuno-tolerance in human early pregnancy.
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ner. Furthermore, IL-10 and TGF-β1 showed a synergic effect. Conclusion IL-10 and TGF-β1, the cytokines of regulatory T cells, suppress the formation and bone resorption function of the osteoclasts.
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Objective To detect the regulation of CXCL16 synthesis and shedding in first-trimester human trophoblasts. Methods Firstly, we analyzed the expression and secretion of chemokine CXCL16 in primary cultured trophoblasts by immunochemical staining and ELISA. Then we determined the soluble and cell-associated CXCLI6 respectively with and without treatments of cytokine IFN-γ, TNF-α, IL-4 and ADAM10 by ELISA. Results Trophoblast expressed and secreted CXCL16 in a stable level. Cytokine IFN-γ induced both synthesis and secretion of CXCL16 significantly ( P <0. 01 ) in trophoblasts. ADAM10 increased the shedding of chemokine domain of CXCL16 from trophoblasts but didn't influence the synthesis of CXCL16 protein in trophoblast. Conclusion IFN-γ and ADAM10 play important roles in production and shedding of transmembrane CXCL16 in first-trimester trophublasts.
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Objective To explore the secretion of chemokine regulated upon activation normal T cell expressed and secreted(RANTES)influenced by the complex microenvironment in the peritoneal cavity of women with endometriosis and investigate chemotaxis of RANTES on the peritoneal monocytes.Methods The contact and non-contact co-culture systems including three target cells of ectopic tissue were established.The three target cells were endometrial stromal cells(ESC),human peritoneal mesothelial cells(HPMC)and monocytes.After collection of the supernatant of co-culture systems,the levels of RANTES were detected by enzyme linked immunosorbent assay(EUSA).Migration of U937 cell,a monocyte line,was detected by chemotaxis assay.Results ESC,HPMC,and U937 cultured alone secreted slight RANTES,(5.0±0.5),(4.0±0.3),and (254±40)ng/L. Compared with the culture of the target cell alone,the levels of RANTES in each co-culture system increased significantly,with the highest level in the contact culture system of E-H-U(2250±96)ng/L. RANTES secretion of non-contact co-culture of three cells were higher than contact co-culture of two cells(P＜0.01):U/E-H(912±93) vs E-H(50±40)ng/L,H/E-U(1201±93) vs E-U(243±192)ng/L,and E/H-U(1519±96) vs H-U(1251±73)ng/L. ESC,HPMC,and ESC-HPMC co-culture improved significantly migration of U937 cells [ number of cell migration respectively(6.0±0.3),(6.2±0.3),(10.0±0.3)×103,P＜0.01],which could be inhibited efficiently by anti-RANTES neutralizing antibody.Condusion The target cells in the peritoneal cavity of patients with endometriosis promote the secretion of RANTES in autocrine and paracrine manners and migration of monocytes.
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The fields of immunology and bone metabolism have matured such that key cellular and molecular mechanisms governing the homeostasis of the individual systems are largely understood. This review summarizes some newly published works in this area and tries to find a noval strategy for the treatment and prevention of diseases related to both bone and immune systems.
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Objective:To investigate into the effects of the BSNXD-treated pharmacological serum on the proliferating cycle and apoptosis of osteoblasts(OBs).Methods: The OBs were isolated by the enzyme-digested assay from calvaria of neonatal BALB/c mice and then cultured.The OBs were treated in vitro with the pharmacological serum in a series of concentration.The apoptosis rate and DNA content of OBs were analyzed by flow cytometry(FCM),and their ultrastructure was evaluated by TEM.Results: The pharmacological serum in 10-20% concentration increased the S-phase and G_(2)/M phase of OB cell cycle,improved the proliferation index(PI,P
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Objective To investigate the roles of matrix metalloproteinase-9, -2 (MMP-9, 2), and tissue inhibitors of metalloproteinase-1,2 (TIMP-1, 2) in pathogenesis of the accretio placenta. Methods The women with the placenta accrete were recruited and the placenta (23) and deciduas tissues (9) after labor were obtained, and the placenta (28) and deciduas (11) from women without the placenta accreta were obtained as control to get, too. The expressions of MMP-9, -2, TIMP-1, 2 in the placental and decidual tissues were analyzed by real-time PCR. Results mRNA expression of MMP-9 in the placenta accreta was (3.21?0.76) copies/?g total RNA, significantly higher (P
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Objective:To study effect of cyclosporin A(CsA) on the prognosis of the abortion prone murine model.Methods:0,5,10,15 mg/kg CsA were injected intraperitoneally to CBA/J female mice at the fourth gestational day which mated to DBA/2 or BALB/c male mice as an abortion prone model and a successful pregnancy model respectively,thereafter the fetus resorption rate,weight of placenta and fetus were surveyed.Results:CsA could reduce the fetal resorption rate of pregnancy loss model significantly,meanwhile the weight of fetus and plancenta did not have notable alteration,but tendency of the weight increase exists.However,CsA did not affect the fetal resorption rate or the weight of fetus and placenta of the successful pregnancy model.Conclusion:CsA injected intraperitoneally at the early stage of pregnancy could reduce the fetal resorption rate of pregnancy loss model to that of normal pregnancy.The weight of fetus and plancenta appeared to have an increase tendency although non significantly.Therefore,CsA seems to be a potential medicine for protecting pregnancy from abortion.
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Objective:To investigate the anti-hCG antibody level in vaginal lavage fluid after vaginal mucosa immunization with the recombinant lactobacillus excreting the human chorionic gonadotropin beta-subunit, hCG?.Methods:The recombinant Lb. casei hCG? was passaged in MRS containing no Erythromycin for 50 generations to get continual secretion. Radio immunol assay(RIA) was used to determine the level of hCG? in the supernatant. Mice of 8 weeks old were immunized with the recombinant continual expressing strain through vaginal inoculation. Two weeks later, level of the hCG? and anti-hCG? IgA antibody in the vaginal lavage was determined by radio-immunology assay and indirect ELISA respectively.Results:The hCG? was secreted by Lb. casei continually. The recombinant hCG? and anti-hCG? IgA antibody in the vaginal lavage were higher than the negative control group(P
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Objective:To evaluate curative impact of booster immunization with paternal lymphocytes on recurrent spontaneous abortors(RSA)with less reaction of primary patermal lymphocyte immunization.Methods:RSA patients with insufficient materno-fetal immuno-recognition were selected by flow cytometry of blocking antibody analysis and immunized with either induced paternal lymphocytes pretreated by IFN-? in vitro to those of anti-CD3-BE and anti-CD4-BE Ab lower than 0% or direct intradermal vaccination with their paternal lymphocytes without IFN-? pretreatment to those of anti-CD3-BE Ab beyond 0%.Reassessment of blocking antibodies was performed at the end of the second immnunization course.Results:Levels of blocking antibodies were significantly raised after the secondary booster immunization in RSA with insufficient materno-fetal immnuno-recognition whose blocking antibodies continuously decreased after being treated by the primary paternal lymphocyte immunization.No improvement of parameters was observed except the blocking effect in patients receiving secondary direct intradermal vaccination treatment.Conclusion:It is necessary for the RSA with insufficient materno-fetal immuno-recognition to experience secondary booster immunization preferably with paternal lymphocytes pretreated by IFN-? in vitro.
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Objective:To probe into if human molecular adjuvant hC3d3 promote the immunogenicity of hCG? antigen to human immuno-competent cells on the basis of fusion protein.Methods:The isolated B cells, the combination of B cells and T cells, PBMC and Raji cells were treated in vitro respectively with 1, 10 and 100 nmol/L hCG?, hCG?-hC3d3 or PWM for 8-12 days. The cell proliferation was determined by incorporation of [3H] thymidine. The Ig levels in the 12-day culture supernatants were measured by indirect ELISA. The Ig-secreting cells in the 10-day cultured lymphocytes were detected by the enzyme-linked immunospot(ELISPOT) assay.Results:It was found that the proliferation of B cells, the combined B and T cells, PBMC and Raji following exposure to hCG?-C3d3 fusion protein was significantly higher than that of hCG? alone. The levels of total Ig the in 12-day culture supernatants of B cell, the combined B and T cells, and PBMC treated with 100 nmol/L hCG?-C3d3 fusion protein were 4-fold, 10-fold and 10.85-fold more than that of hCG? alone. The Ig-secreting cells were significantly increased after treated with hCG?-C3d3 fusion protein compared to the hCG? alone.Conclusion:The human molecular adjuvant hC3d3 improves the immunogenicity of hCG? in human immuno-competent cells if fused to the antigen.
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Objective:To induce an anti-hCG? antibody response in reproductive tract and the peripheral by oral inoculation with a recombinant alive Lactobacilli expressing hCG? in different strains of mouse.Methods:The 108,109,1010 Lb.hCG?, a recombinant Lactobacillus expressing hCG?, were inoculated orally into 6-8-week-old female BALB/c and C57BL/6 mice. The immunized mice were all boosted with the same dose at interval of 3 weeks. An indirect ELISA was used to determine anti-hCG? IgG and IgA antibodies in vaginal lavage and serum that were got at the end of 2-8 weeks after the primary immunization. The B and T lymphocytes of spleen, uterus and vagina were prepared respectively, and their proliferation was analyzed by flow cytometry. The numbers of the cells secreting anti-hCG? IgG or IgA antibody were detected by ELISpot.Results:Inoculation of 109 or 1010 alive Lb.hCG? could induce similar response while 108 induce a lower attitude of response. The antiserum of either 109 or 1010 Lb. hCG? immunization after booster could neutralized more than 100 ng/ml of hCG antigen both in BALB/c and C57BL/6 mice. The B cell proliferation of the reproductive mucosa was more significantly higher than that of spleen in BALB/c mice(P
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Objective:To probe into effects of Mifepristone on the Th1/Th2 balance at materno-fetal interface.Methods:Sixty three healthy women in early pregnancy were randomized into two groups,30 women used mifepristone 200 mg,and 33 women as control.Their decidual tissue was collected by curettage in 48 hours of taking the drug.The Th1/Th2 cytokine expressions in decidua were analyzed by immunohistochemistry.Results:In the decidual tissue of normal early pregnancy,there were significantly higher densities of Th2 cytokines,IL-4 and TGF-?2,and lower density of Th1-type cytokine,IFN-? and IL-2.After having treated by Mifepristone,the decidual tissue appeared to significantly higher densities of Th1-type cytokines(IL-2, IFN-?).Conclusion:Our findings suggest that Mifepristone can up-regulate expression of the Th1-type cytokines,and cause a shift towards Th1 bias,which leads to abortion.
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Objective:Investigating the expression of HIV-1 co-receptors CXCR4,CCR5 and chemoking SDF-1 in human placentae and trophoblasts is to explore the mechanism of in-utero transmission of human immunodeficiency virus type 1(HIV-1).Methods:Using semi-quantitative reverse transcriptase-polymerase chain reaction,detected the transcripts of CXCR4,CCR5 in placenta tissues and trophoblasts.Immunocytochemistry and immunohistochemistry analysis revealed the expression of CXCR4,CCR5 in primary cultured first trimester trophoblasts and villous tissue.Also the expression of SDF-1 in villi of first trimester,and the presence of SDF-1 in the culture of isolated trophoblasts were examined by in situ hybridization,immunohistochemistry and enzyme-linked immunosorbent assay.Results:CXCR4 and CCR5 mRNA were detected in placentae of various gestational phases and in first trimester trophoblasts.However,primary cultured trophoblasts were found to be reactive only with antibodies against CXCR4.In the villous tissue sections,CXCR4 was found in trophoblasts,while CCR5 in stromal cell and/or Hofbauer cell.First trimester trophoblasts expressed SDF-1 strongly and secreted SDF-1 in vitro.Conclusion:CXCR4 and CCR5 were expressed in placentae.Hence,they maybe involved in in-utero transmission of HIV-1 as co-receptors.However,SDF-1 expressed in trophoblasts may protect fetus from being invaded by X4-HIV-1.R5-HIV-1 may infect CCR5+ stromal cells and/or Hofbauer cells through placental disruptions.
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PurposeTo exploit the relationship between the history of abortion and the infection of viral hepatitis C in the pregnant women.MethodsRisk factors for hepatitis C virus(HCV) infection were analyzed by Chi-square and a multiple logistic regression analysis in 2 783 pregnant women from Shanghai Punan hospital in 1995- 1998.ResultsThe results showed that the following factors associated significandy with anti-HCV seropositive rate:number of abortion,anti-HCV positive of spouse. In the logistic regression analysis the prevalence rate of HCV in pregnant women who were laid-off their work was 7.37 %, which was higher than that of other occupational pregnant women. There was no relationship between the HCV infection and age,marital status,history of injection, operation, hospitalization and the drug abuse.ConclusionsAbortion is a risk factor of HCV infection in pregnant women. The positive rate of anti-HCV is increased with the number of abortion. Strengthening thesterilization and disinfection of instruments during the abortion operation is necessary.
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Objective:In the past study, a plasmid of pcDNA3 hCG? C3d3 had been constructed. It was suggested that the fusing protein be expressed by the pcDNA3 hCG? C3d3 plasmid both in the transient expression system in COS 7 cells, and the stable expression system in CHO cells. In the present research, it would be testified that the C3d molecular adjuvant could improve the immunogenicity of the hCG? DNA immunization or not. Methods:BALB/C mice of 6 weeks old were immunized intramuscularly two times at interval of 3 weeks by the plasmid pcDNA3, pcDNA3 hCG?, pcDNA3 hCG? C3d3 at dosage of 5, 10, 20 pmol, respectively. The anti hCG? antibody titers were determined by indirect ELISA in 6 weeks of last immunization. Results:The result showed that the C3d molecular adjuvant could increase significantly the titer of anti hCG? antibody in a dose dependent manner after DNA immunization.Conclusion:The C3d molecular adjuvant does improve the humoral immunity of the hCG? DNA immunization, which would be helpful for technical progress and clinical trial of the contraceptive vaccine. [
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Objective:To explore the role of CD86 costimulation in inducing Th2 bias at maternal-fetal interface and the relationship to outcomes of gestation.Methods:Pregnant DBA/2J mated CBA/J mice with a high embryo resorption rate from 20% to 30% and BALB/C mated CBA/J mice with low embryo resorption rates were studied,with rat anti-murine CD86 mAb administered intraperitoneally at the dosage of 100 ?g,at the time of implantation(day 4) and on the following days(6,8,10) of gestation.The competitive semiquantity RT-PCR and ELISA was applied to analysis of the transcription and expression of Th type-1/Th type-2 cytokines at the maternal-fetal interface at day 9 or day 14 of gestation respectively.The embryo resorption rate was counted at day 14 of gestation.Results:In the model of normal pregnancy,blockade of CD86 costimulation had no significant effects on the original Th2 bias at the maternal-fetal interface,and the outcomes of gestation had not changed significantly.While in the model of abortion-prone,blockade of CD86 costimulation successfully induced a Th2 bias at maternal-fetal interface.Therefore,the embryo resorbing rates decreased significantly.Conclusion:Blocking the CD86 costimulation at the early stage of the abortion-prone pregnancy could recover the physiological balance of Th1/Th2 at maternal-fetal interface and induce the maternal-fetal immune tolerance.
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Objective:To investigate the expression of chemokine receptor in eupotic and ectopic endometrial tissues of women with endometriosis, and in endometrium of women without endometriosis.Methods:Normal endometrium, eutopic endometrium and endometriotic tissues were obtained from patients with endometriosis at laparoscopy. Total RNA was then extracted using the TRIzol reagent. The expression of chemokine receptors in these tissues were analyzed by way of semi-quantitative reverse transcriptase-polymerase chain reaction.Results:Compared to normal endometrium, the eutopic endometrium expressed significantly more CCR6, CCR8, CCR9 and CX3CR1(P