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OBJECTIVE@#To investigate the correlation of inducible co-stimulatory molecules (ICOS) with mesenteric vascular endothelial- mesenchymal transition (EndMT) and sclerosis in spontaneously hypertensive rats (SHR).@*METHODS@#Twenty 4-week-old WKY rats and 20 SHRs of the same strain were both randomly divided into 4 groups for observation at 4, 6, 10 and 30 weeks of age. ICOS expression frequency in rat spleen CD4+T cells was analyzed using flow cytometry, and the expressions of ICOS, VE-cad, α-SMA and Col3 mRNA in rat mesentery were detected by RT-PCR. The distributions of ICOS, IL-17A and TGF-β in rat mesentery were detected by immunohistochemistry. The levels of IL-17A and TGF-β in rat plasma were measured using ELISA. The morphological changes of rat mesenteric vessels were observed with Masson staining. Spearman or Pearson correlation analyses were used to evaluate the correlation between ICOS expression and the expressions of the markers of vascular EndMT and sclerosis.@*RESULTS@#Compared with the control WKY rats, the SHRs began to show significantly increased systolic blood pressure and ICOS expression frequency on CD4+T cells at 6 weeks of age (P < 0.05). In the SHRs, the mRNA and protein expressions of ICOS, α-SMA, Col3, IL-17A and TGF-β in the mesentery were significantly higher than those in control group (P < 0.05), while the mRNA and protein expressions of VE-cad started to reduce significantly at 10 weeks of age (P < 0.05). The plasma levels of IL-17A and TGF-β were significantly increased in SHRs since 6 weeks of age (P < 0.05) with progressive worsening of mesenteric vascular sclerosis (P < 0.05). ICOS mRNA and protein expression levels in the mesenteric tissues of SHRs began to show positive correlations with α-SMA and Col3 expression levels and the severity of vascular sclerosis at 6 weeks of age (P < 0.05) and a negative correlation with VE-cad expression level at 10 weeks (P < 0.05).@*CONCLUSION@#ICOS play an important pathogenic role in EndMT and sclerosis of mesenteric vessels in essential hypertension by mediating related immune responses.
Subject(s)
Rats , Animals , Rats, Inbred SHR , Rats, Inbred WKY , Hypertension , Interleukin-17 , Sclerosis/pathology , Transforming Growth Factor beta , Mesentery/pathology , RNA, Messenger/metabolism , Blood PressureABSTRACT
Immune checkpoint consists of inhibitory and stimulatory molecules. Drugs blocking inhibitory checkpoint programmed cell death receptor-1 (PD-1) and cytotoxic T lymphocyte-associated molecule-4 (CTLA-4) are currently utilized for wide variety of human cancers. Agonists of stimulatory checkpoints such as GITR, OX40, 4-1BB, ICOS, CD40 and STING are undergoing critical clinical trials. Immune checkpoint agonists that affect stimulatory checkpoint molecules develop rapidly, and immune agonist antibodies thus represent an important approach for solid tumor treatments.
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Background Osteoclast stimulatory transmembrane protein (OC-STAMP) is involved in silicosis fibrosis induced by silicon oxide (SiO2) exposure. Its role in silicosis fibrosis by inducing ferroptosis of alveolar type II epithelial cells and its related mechanism remain unclear. Objective To explore the effect and possible mechanism of OC-STAMP on ferroptosis of alveolar type II epithelial cells and silicosis fibrosis in rats under SiO2 exposure. Methods Twenty male Wistar rats of SPF grade were randomly divided into two groups: control (Sham) group and SiO2 group, 15 rats in each group. Rats in the SiO2 group were given 1 mL of 50 mg·L−1 SiO2 suspension at one time through the non-exposed intratracheal instillation method to establish an animal model of silicosis, and rats in the Sham group were give 1 mL of 0.9% sodium chloride solution in the same way. Rats were sacrificed after 8 weeks. Samples of lung tissue were fixed in glutaraldehyde or paraformaldehyde for observing ultrastructure of mitochondria by transmission electron microscopy; HE, Masson, VG, and Prussian blue were used to observe changes in lung tissue structure and iron deposition. The expression level of OC-STAMP and the degree of lung fibrosis were evaluated by immunohistochemistry and immunofluorescence. The expression level of OC-STAMP in rat lung tissue was detected and the transfection effect of OC-STAMP was verified by real-time fluorescence quantitative polymerase chain reaction (RT-PCR). Overexpression (OCS group) and inhibition expression (SI-OC group) models were constructed by OC-STAMP plasmid and OC-STAMP small interfering RNA (siRNA) transfection to cultured MLE-12 cells, respectively. The relative expression levels of glutathione peroxidase 4 (GPX4), solute carrier family 7 member 11 (SLC7A11), and other proteins in lung tissue and MLE-12 were detected by Western blotting. Results The results of HE, Masson, and VG staining showed that the silicosis modeling was successful after 8 weeks of SiO2 exposure. The immunofluorescence results showed that OC-STAMP and ATP binding cassette subfamily A member 3 (ABCA3) co-localized in alveolar type II epithelium. The immunohistochemical results showed that the levels of OC-STAMP and collagen I in the SiO2 group were significantly higher than those in the Sham group (P<0.01). The RT-PCR results showed that the OC-STAMP mRNA in the lung tissue of the SiO2 group was significantly higher than that of the Sham group (P<0.01). The Prussian blue staining in the lung tissue of the SiO2 group showed positive brownish-yellow particles. Compared with the Sham group which showed normal mitochondrial structure, the mitochondrial structure was generally swollen and the mitochondrial cristae dissolved and disappeared in the SiO2 group by transmission electron microscope observation. The Western blotting results showed that the expression levels of SLC7A11 and GPX4 both decreased in the lung tissue of the SiO2 group (P<0.05, P<0.01), and the expression level of Vimentin increased (P<0.01). In the transfected MLE-12 cells, compared with the Sham group, the expression levels of SLC7A11 and GPX4 in the OCS group were significantly reduced (P<0.05, P<0.01). Conclusion OC-STAMP may affect the expression of proteins related to ferroptosis, and promote lung fibrosis induced by SiO2 exposure.
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As a co-stimulatory blocker against CD28 receptor, belatacept has been approved and applied to the treatment of rejection in organ transplantation in Europe and America. Belatacept has been proven to outperform calcineurin inhibitor (CNI) in improving the long-term survival rate of recipients and grafts, and enhancing graft function. Nevertheless, it might cause a high incidence of rejection. To resolve this issue, transplant workers have attempted to optimize belatacept immunosuppressive regimen and achieved good clinical efficacy. Although belatacept has been proven to exert poor effect on memory T cells, it has potential value in exploring new co-stimulatory molecular targets to optimize immunosuppressive regimes due to its specificity for immune cells and mild adverse effects. In this article, the advent of co-stimulatory blocker, clinical efficacy and application of belatacept, and the causes of belatacept-resistant rejection were reviewed.
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Abstract The mealybug, Phenacoccus solenopsis Tinsley (Hemiptera: Pseudococcidae) is a cotton pest widespread in several cotton growing regions of Brazil, particularly in the semi-arid region of southwestern Bahia. The impact of kaolin on survival, development and reproduction of P. solenopsis was evaluated in the laboratory. The experiment was developed in a completely randomized design with two treatments: immature or newly emerged adults of P. solenopsis sprayed with kaolin and fed with cotton leaf discs treated with kaolin suspension (with kaolin) (T1) and immature or newly emerged adults of P. solenopsis sprayed with distilled water and fed with cotton leaf discs treated with distilled water (without kaolin) (T2). The kaolin suspension shortens the life cycle, increases the reproductive potential and population growth of the cotton mealybug, P. solenopsis and, therefore, it should be used with caution on cotton plants in regions with a history of occurrence of this pest.
Resumo A cochonilha Phenacoccus solenopsis Tinsley (Hemiptera: Pseudococcidae) é uma praga de algodão, amplamente, difundida em várias regiões produtoras do Brasil, particularmente, na região semiárida do sudoeste da Bahia. O impacto do caulim na sobrevivência, desenvolvimento e reprodução de P. solenopsis foi avaliado em laboratório. O delineamento experimental foi inteiramente casualizado com dois tratamentos: imaturos ou adultos recém-emergidos de P. solenopsis alimentados com discos foliares de algodão tratados com caulim (com caulim) e pulverizadas com este produto (T1) e imaturos ou adultos recém-emergidos de P. solenopsis alimentados com discos foliares de algodão tratados com água destilada (sem caulim) e pulverização dos insetos com água destilada (T2). A suspensão do caulim encurtou o ciclo de vida, aumentou o potencial reprodutivo e o crescimento populacional da cochonilha-do-algodoeiro, P. solenopsis e, portanto, deve ser utilizado com cautela em plantas de algodão em regiões com histórico de ocorrência desta praga.
Subject(s)
Animals , Hemiptera , Reproduction , Brazil , Gossypium , KaolinABSTRACT
The normal immune system has the ability to distinguish between "self" and "non-self". Because of its dynamic balance of "immune activity-immune tolerance", it will produce immune response to the non-self antigen, but with no response or weak response to the self-antigen. However, if the balance was broken, T cell in the abnormal immune activation state will respond continually to the self-antigen, with an abnormal immune response, which caused autoimmune disease. Pathologically, "invalid" immune recognition and immune response become the main causes for autoimmune diseases. Co-stimulatory molecule is an important link between Attach antigen presenting cells（APC） and immune cells (T cell and B cell). Studies have proved that excessive co-stimulation and/or insufficient co-inhibition could cause detect of self-tolerance and induce autoimmunity. Although co-stimulatory and co-inhibitory pathways have a significant impact on all ADS, this paper focuses on their effect on two systemic autoimmune diseases [systemic lupus erythematosus （SLE） and rheumatoid arthritis（RA）] and two organ-specific autoimmune diseases [multiple sclerosis （MS） and type 1 diabetes （T1DM）], in order to discuss the pathogenesis and relationship between co-stimulatory molecules and autoimmune diseases.
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Objective: To generate mouse B7-2 gene RNAi lentivirus and study its interference effects on B7-2 expression and T lymphocytes proliferation induced by dendritic cells. Methods: Three sequences specific targeting B7-2 gene and one non-specific sequence were respectively synthesized, and inserted into lentiviral vector, then the recombinant vectors were sequencing. 293 T cells were co-transfected with lentiviral expression plasmid and packaging plasmids to produce recombinant lentivirus which titre was checked according to the expression level of green fluorescent protein ( GFP). Bone marrow cells from C57 BL/6 mice were isolated to differentiate into DCs at the present of GM-CSF, IL-4 and LPS for 48 h, then morphology and phenotypic was identified. DCs were infected by recombinant RNAi lentivirus and then the efficiency of infection and the expression of B7-2 on the surface of DCs were detected by flow cytometry. Effects on the proliferation of T cells were detected by co-culturing with DCs which were infected by B7-2 RNAi lentivirus and murine spleen T cells in vitro. Results: DNA sequencing confirmed that three B7-2 RNAi and one non-specific recombinant lentiviral transfer plasmids were successfully constructed, the titer of recombinant lentivirus was ( 2-4) × 108 TU/ml. The recombinant lentivirus could effectively infect DC and inhibit the expression of B7-2. After the B7-2 recombinant lentivirus infection, the ability of DCs to stimulate the proliferation of T cells decreased obviously ( P＜0. 05). Conclusion: The lentiviral B7-2 gene RNAi vector can effectively silence the expression of B7-2 on the surface of DCs and inhibit the proliferation effect of T cells induced by DCs.
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Objective To investigate the clinical significance of postoperative stimulated thyroglobulin ( ps-Tg) detection in the patients with differentiated thyroid carcinoma ( DTC) 131 I therapy. Methods Totally 73 cases with DTC who were hospitalized from March 2009 to March 2018 with complete treatment and follow-up data were selected as the study subjects. The cases were divided into three groups by 131I scanning image, no metastasis group (32 cases), lymph node metastasis group (31 cases), and distance metastasis group ( 10 cases ) . The levels of stimulated thyroglobulin ( s-Tg ) in serum were detected within one week before 131 I ablation for thyroid remnant and metastasis after surgical treatment. The 131I -whole body scan (131I -WBS) was performed at 3 to 5 days after 131I treatment and these cases were followed-up for 8-12 months. Results There was not statistically significant difference in ps-Tg between no metastasis group and lymph node metastasis group before 131I treatment(P>0.05). The ps-Tg in distance metastasis group was significantly higher than that in patients without metastasis and lymph node metastasis(Z= -3.810、 Z= -3.371, P<0.05). Before treatment with 131I ablation metastasis, there was not statistically difference in s-Tg among 3 groups(H=11.764, P<0.05). The area under the ROC curve of ps-Tg and s-TG diagnostic DTC distant metastasis rate before treatment of ablation for thyroid remnant was 0.903, and the accuracy were 90.48％. The area under the ROC curve of diagnosis of DTC distant metastasis and lymph node metastasis before ablation for thyroid metastasis was 0. 817 and 0.644, and the accuracy was 88.10％ and 65.08％, respectively. Conclusions The accuracy of diagnosing DTC distant metastasis of ps-Tg before 131 I ablation for thyroid remnant is superior to ps-Tg before ablation forthyroid metastasis. The level of s-Tg diagnosis of DTC lymph node before 131 I ablation metastasis precede ps-Tg before 131 I ablation for thyroid remnant
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Objective:To explore the effects of suppress or enhancer of lin-12-like(Sel1L) on differentiation and function of bone marrow-derived dendritic cells.Methods:To generate conditional knockout mice by the Cre-Loxp recombination system.ELISA and Real-time fluorescence quantitative PCR(RT-PCR) was used for analyzing the protein levels and mRNA levels of IL-6/IL-12 in BMDCs.The protein levels of Sel1L in BMDCs were detected by Western bolt.The expression of CFSE,CD80,CD86,MHC-Ⅰ,MHC-Ⅱon BMDCs and the capability in priming OVA specific CD4+T cells proliferation were analyzed by the flow cytometry.Results:The deficiency of Sel1L decreases the proliferation of DCs during its differentiation,up-regulates the secretion of IL-6,IL-12 and the expression of MHC-Ⅰ.Notably,Sel1L-null DCs was failed to up-regulate MHC-Ⅱexpression and dramatically impaired their ability to prime OVA323-339specific CD4+T cell.Conclusion:The deletion of Sel1L can reduce the proliferation of BMDCs and down-regulate its ability in priming the proliferation of OVA specific CD4+T cells.
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Objective To investigate the effects of water extracts of Lepidium meyenii walp (LMWE) collected from two different places in Xinjiang on the maturation and function of dendritic cells (DCs) and to evaluate their roles in immunoregulation. Methods Water extracts of LMWE growing in Tashikuergan County (Ta xian) and A La gou of Xinjiang were prepared and named as LMWE-T and LMWE-A,respectively. Bone marrow-derived DCs and splenocytes isolated from C57BL/6 mice were treated with different concentrations of polysaccharide extracts from LMWE-T/A. Effects of LMWE-T/A on the per-centage and apoptosis of DC,expression of co-stimulatory molecules and secretion of cytokines were detected by flow cytometry and ELISA. MTT assay was used to analyze the proliferation of splenocytes. Changes in the functions of DC were evaluated by mixed lymphocyte reaction(MLR). Results LMWE-T/A had no in-fluence on the percentage and viability of DC. Expression of CD40 and CD86,and secretion of TNF-α,IL-12p40 and IFN-γ were significantly increased by LMWE-T/A treatment in a dose-dependent manner. LMWE-T/A treatment enhanced the functions of DCs and also dose-dependently promoted the proliferation of splenocytes. LMWE-A was more effective than LMWE-T in promoting CD86 expression,IFN-γ secretion and splenocyte proliferation. Pretreatment with TAK-242,an inhibitor of Toll-like receptor 4(TLR4),could sig-nificantly inhibit the regulatory effects of LMWE-T/A on CD40 expression and the secretion of TNF-α and IL-12p40. Conclusion This study shows that LMWE could promote the maturation of DC through TLR4 signaling pathway,enhance the functions of DC without side effects on DC survival,and increase the prolif-eration of splenocytes,indicating that LMWE has a potential immunopotentiating effect. LMWE-A has better effects than LMWE-T on immune enhancement.
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Objective To study the impacts of B7-H3 molecule on the proliferation of T lymphocytes in different activation conditions and on the secretion of relevant cytokines.Methods Peripheral blood samples were collected from healthy subjects to separate peripheral blood mononuclear cells (PBMC) by Ficoll density gradient centrifugation.T lymphocytes were isolated from some of the PBMCs and purified with T Cell Enrichment Kit.PBMC and purified T lymphocytes were activated by anti-CD3/CD28 monoclonal antibodies (McAb) in vitro.Flow cytometry analysis was used to detect the expression of CD28 and CTLA-4 on T lymphocytes at different time points for further analyzing the activation states of T lymphocytes.On this basis, human B7-H3-Fc fusion protein was added into the mixed co-cultivation system on days 0, 1, 2, 3 and 4, and Hu-Fc fusion protein was used as isotype control.CCK-8 method was performed to detect the proliferation of T lymphocytes in each group.ELISA method was used to detect the secretion of cytokines (IL-2, IL-10 and IFN-γ) and to analyze the immune responses induced by stimulating T lymphocytes at different states of activation with B7-H3.Results B7-H3 molecule significantly inhibited the quiescent T lymphocytes from secreting IL-2 and IL-10, but had no significant impact on IFN-γ secretion.Moreover, it significantly promoted the activated T lymphocytes to secret IL-2 and IFN-γ, but had no obvious impact on IL-10 secretion.Results of the cell proliferation assay showed that B7-H3 molecule inhibited the in vitro proliferation of T lymphocytes in the PBMC, but had no obvious impact on purified T lymphocytes.ConclusionThe regulatory effects of B7-H3 molecule on the immune functions of T lymphocytes vary with the activation states of T lymphocytes.
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B7-H3,a newly discovered co-stimulatory regulatory protein member of the B7 family.Its mRNA is ubiquitously expressed in a wide spectrum of tissues.As a co-stimulatory or co-inhibitory signal molecule which can regulate immune response,B7-H3 plays an important role in the immune system.Besides,B7-H3 can be also involved in cancer progression via non-immunological.Recently,the aberrant expression of B7-H3 has been described in various malignancies,and significantly correlated with poor prognosis and cancer progression.Therefore,B7-H3 is considered as an early diagnostic and prognostic marker and therapeutic target for tumors,but the specific molecular mechanisms of B7-H3 regulation are poorly understood.The immune and gene therapy of tumor by target B7-H3 has made some progress.
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Objective To study the therapeutic effect of anti-CD80 bivalent antibody on mouse lu-pus nephritis and to explore the possible molecular mechanism. Methods A mouse model of lupus nephritis was established through intraperitoneal injection of 0. 5 ml of pristine in female C57BL/6J mice. Appearance of urinary protein and significantly increased levels of peripheral antinuclear antibody ( ANA) and anti-doub-le-stranded DNA ( anti-dsDNA) antibody in the fourth month after injection indicated that the mouse model was established successfully. Then the mice were divided into two groups including anti-CD80 bivalent anti-body intervention group (injected with 200μg of anti-CD80 bivalent antibody at day 1, 3, 5, 8 and 15, fol-lowed by three times of injection with an interval of one month) and model group ( injected with the same protein using the same strategy). A normal control group was set up accordingly. Albustix test paper was used to monitor the dynamic changes in mouse urinary protein. Flow cytometry was used to analyze the acti-vation of immune-related cells in spleen. Levels of autoantibodies ( ANA and anti-dsDNA) and levels of IFN-γ and IL-4 in serum were detected by indirect immunofluorescence assay. Renal tissue samples were an-alyzed with hematoxylin and eosin ( HE) staining and immunocomplex ( IC) assay. Results Urinary pro-tein level of the anti-CD80 bivalent antibody intervention group was significantly lower than that of the model group (P<0. 05). Activated macrophages, dendritic cells, neutrophils and B cells in spleen tissues of the anti-CD80 bivalent antibody intervention group were significantly less than those of the model group ( P<0. 05), and the numbers of CD4+ and CD154+ T cells were significantly less than those of the model group (P<0. 05). Positive rates and titers of ANA and dsDNA in serum samples of the intervention group were lower than those of the model group (P<0. 05). Levels of IFN-γand IL-4 in serum samples of the interven-tion group were decreased as compared with those of the model group (P<0. 05). HE staining and immuno-fluorescence assay showed that glomerular inflammatory injury and necrosis were alleviated and kidney im-mune complex deposition was reduced after anti-CD80 bivalent antibody intervention. Conclusion Anti-CD80 bivalent antibody specifically binds to the CD80 molecule on antigen presenting cell surface, blocks the CD80/CD28 co-stimulatory signaling pathway and down-regulates the body′s immune response, which al-leviates and reverses the lupus-like nephritis-induced pathological damages in mice.
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Objective To study the therapeutic effect of anti-CD80 bivalent antibody on mouse lu-pus nephritis and to explore the possible molecular mechanism. Methods A mouse model of lupus nephritis was established through intraperitoneal injection of 0. 5 ml of pristine in female C57BL/6J mice. Appearance of urinary protein and significantly increased levels of peripheral antinuclear antibody ( ANA) and anti-doub-le-stranded DNA ( anti-dsDNA) antibody in the fourth month after injection indicated that the mouse model was established successfully. Then the mice were divided into two groups including anti-CD80 bivalent anti-body intervention group (injected with 200μg of anti-CD80 bivalent antibody at day 1, 3, 5, 8 and 15, fol-lowed by three times of injection with an interval of one month) and model group ( injected with the same protein using the same strategy). A normal control group was set up accordingly. Albustix test paper was used to monitor the dynamic changes in mouse urinary protein. Flow cytometry was used to analyze the acti-vation of immune-related cells in spleen. Levels of autoantibodies ( ANA and anti-dsDNA) and levels of IFN-γ and IL-4 in serum were detected by indirect immunofluorescence assay. Renal tissue samples were an-alyzed with hematoxylin and eosin ( HE) staining and immunocomplex ( IC) assay. Results Urinary pro-tein level of the anti-CD80 bivalent antibody intervention group was significantly lower than that of the model group (P<0. 05). Activated macrophages, dendritic cells, neutrophils and B cells in spleen tissues of the anti-CD80 bivalent antibody intervention group were significantly less than those of the model group ( P<0. 05), and the numbers of CD4+ and CD154+ T cells were significantly less than those of the model group (P<0. 05). Positive rates and titers of ANA and dsDNA in serum samples of the intervention group were lower than those of the model group (P<0. 05). Levels of IFN-γand IL-4 in serum samples of the interven-tion group were decreased as compared with those of the model group (P<0. 05). HE staining and immuno-fluorescence assay showed that glomerular inflammatory injury and necrosis were alleviated and kidney im-mune complex deposition was reduced after anti-CD80 bivalent antibody intervention. Conclusion Anti-CD80 bivalent antibody specifically binds to the CD80 molecule on antigen presenting cell surface, blocks the CD80/CD28 co-stimulatory signaling pathway and down-regulates the body′s immune response, which al-leviates and reverses the lupus-like nephritis-induced pathological damages in mice.
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Abstract The mealybug, Phenacoccus solenopsis Tinsley (Hemiptera: Pseudococcidae) is a cotton pest widespread in several cotton growing regions of Brazil, particularly in the semi-arid region of southwestern Bahia. The impact of kaolin on survival, development and reproduction of P. solenopsis was evaluated in the laboratory. The experiment was developed in a completely randomized design with two treatments: immature or newly emerged adults of P. solenopsis sprayed with kaolin and fed with cotton leaf discs treated with kaolin suspension (with kaolin) (T1) and immature or newly emerged adults of P. solenopsis sprayed with distilled water and fed with cotton leaf discs treated with distilled water (without kaolin) (T2). The kaolin suspension shortens the life cycle, increases the reproductive potential and population growth of the cotton mealybug, P. solenopsis and, therefore, it should be used with caution on cotton plants in regions with a history of occurrence of this pest.
Resumo A cochonilha Phenacoccus solenopsis Tinsley (Hemiptera: Pseudococcidae) é uma praga de algodão, amplamente, difundida em várias regiões produtoras do Brasil, particularmente, na região semiárida do sudoeste da Bahia. O impacto do caulim na sobrevivência, desenvolvimento e reprodução de P. solenopsis foi avaliado em laboratório. O delineamento experimental foi inteiramente casualizado com dois tratamentos: imaturos ou adultos recém-emergidos de P. solenopsis alimentados com discos foliares de algodão tratados com caulim (com caulim) e pulverizadas com este produto (T1) e imaturos ou adultos recém-emergidos de P. solenopsis alimentados com discos foliares de algodão tratados com água destilada (sem caulim) e pulverização dos insetos com água destilada (T2). A suspensão do caulim encurtou o ciclo de vida, aumentou o potencial reprodutivo e o crescimento populacional da cochonilha-do-algodoeiro, P. solenopsis e, portanto, deve ser utilizado com cautela em plantas de algodão em regiões com histórico de ocorrência desta praga.
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AIM:To explore the effect of MCSF and its receptor on the response of ovarian stimulation by comparing the expression of macrophage colony stimulating factor (M-CSF) and its receptor mRNA in luteinized granulosa cells in patients after using combinant human follicle stimulating hormone.METHODS:Ninety-six patients with polycystic ovary syndrome (PCOS) and 157 patients with non-PCOS underwent in vitro fertilization were divided into four groups,i.e.the PCOS fast and slow reaction group,and the non PCOS fast and slow reaction group,according to their response to recombinant human follicle stimulating hormone (r-FSH).Luteinized granulosa cells were then collected after mature follicular puncture.SYBR Green quantitative RT-PCR method was used to detect the expression of M-CSF,M-CSFR and GAPDH in the mRNA gene of the granulosa cells samples.The relative quantity of these genes were determined by comparing the threshold value (CT value of the target gene subtract CT value of housekeeping gene).The difference of gene expression between two groups was compared by t test,and Spearman correlation analysis was used to describe the data relationships.RESULTS:No significant difference was observed in the use of r-FSH among the different groups (P ＞ 0.05).Neither was there any significant difference in mRNA quantity of M-CSF or M-CSFR between the entire PCOS and non PCOS patients (P ＞ 0.05).After grouping,no significant difference was observed between any two groups in the expression of M-CSF (P ＞ 0.05).The expression of M-CSFR in PCOS slow response group was significantly lower than that of PCOS fast response group (P =0.006).Meanwhile,the Spearman analysis showed that the correlation between the quantification of M-CSFR mRNA and the days of r-FSH in PCOS group was statistically significant (P =0.023);the correlation coefficient was 0.511.CONCLUSION:The slow response to ovarian stimulation in PCOS patients is possibly related to the reduction of granulocyte MCSFR expression.The M-CSF and its receptor may be involved in the ovarian stimulation response process.
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Objective To investigate the therapeutic effects of rapamycin (RAPA) on concanavalin A (ConA)-induced acute autoimmune hepatitis in a mouse model and to analyze the possible mechanism.Methods Thirty eight-week-old female C57BL/6 mice were randomly divided into three groups including control group,ConA model group and ConA + RAPA treatment group.The levels of alanine transaminase (ALT) and aspartate aminotransferase (AST) in serum samples were measured after injection of mice with ConA for 24 hours for assessing the liver function.Hematoxylin-eosin (HE) staining was used to observe the hepatic pathological changes in mice.Splenocytes were harvested 24 h after ConA injection for the detection of the percentages of splenic DCs,CD4+T,CD8+T and CD4+ Foxp3+ Treg cells as well as the expression of co-stimulatory molecules (CD40,CD80 and CD86) on DCs by using flow cytometry.Results The levels of ALT and AST in mice from the RAPA treatment group were significantly lower than those of the ConA model group.Results of the HE staining assay showed that the liver damages in RAPA treated mice were less severe than those in mice from the ConA model group.Compared with mice form the ConA model group,those treated with RAPA showed decreased percentages of splenic CD4+ and CD8+ T cells,inhibited expression of CD80 and CD86 on splenic DCs,but increased percentages of splenic CD4+ Foxp3+ Treg cells.No statistically significant differences in the percentages of splenic DCs and the expression of CD40 were observed between the RAPA treatment group and the ConA model group.Conclusion The immunosuppressive effects of RAPA on mice with ConA-induced hepatitis might be achieved through the regulation of immune cells including DCs and T cells in spleen tissues.This study might pave the way for further investigation on the prevention and treatment of autoimmune hepatitis.
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Objective:To construct lentiviral vector specific for mouse B7-1 RNA interference and study lentivirus-mediated B7-1 gene silencing effects in L929 fibroblast cells.Methods:Three candidate sequences for B7-1 RNAi selected from coding sequence of mouse B7-1 transcription were used to design short hairpin RNA ( shRNA ) templates and then cloned into lentiviral expression plasmid followed with correctness identification of inserted sequence by DNA sequencing.Recombinant lentivirus were prepared by co-transfecting lentiviral expression vector and packaging plasmids into 293T cells.Then the resulting culture supernatant containing infectious lentiviral particles was pooled and centrifuged via ultra-centrifugation.Infectious titer of the preparations was determined by detecting the expression of GFP in 293T cells after transfected by lentivirus.Cultured L929 cells were transfected with lentivirus to deter-mine transduction efficiency and silencing efficacy of B7-1 expression by flow cytometry.Transducted L929 cells were then screened using puromycin to generate stable cell clones followed by flow cytometry analysis of GFP and B7-1 expression.A mixed reaction system consisting of stable B7-1 silencing L929 cells and mouse splenic T cells was used to analyze ability of the established cell line to trigger T cells proliferation.Results: Lentiviral expression vector for mouse B7-1 RNAi was correctly constructed with inserted sequences as designed.Recombinant RNAi lentivirus were prepared with titers ranging (3-5) ×108 TU/ml and efficacy to mediate GFP transgene expression and B7-1 silencing.B7-1 expression and the ability to trigger T cells proliferation of stable L929 cells were suppressed significantly ( P<0.05 ).Conclusion: We generated lentiviral vector specific for mouse B7-1 RNAi with high performance of transduction efficiency as well as B7-1 silencing efficacy and the recombinant RNAi lentivirus can mediated stable B7-1 gene silencing in L929 cells and inhibition of T cells proliferation induced by B7-1/CD28 co-stimulatory signal.
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CD28 is a primary co-stimulatory receptor that is essential for successful T cell activation, proliferation, and survival. While ubiquitously expressed on naive T cells, the level of CD28 expression on memory T cells is largely dependent on the T-cell differentiation stage in humans. Expansion of circulating T cells lacking CD28 was originally considered a hallmark of age-associated immunological changes in humans, with a progressive loss of CD28 following replicative senescence with advancing age. However, an increasing body of evidence has revealed that there is a significant age-inappropriate expansion of CD4⁺CD28⁻ T cells in patients with a variety of chronic inflammatory diseases, suggesting that these cells play a role in their pathogenesis. In fact, expanded CD4⁺CD28⁻ T cells can produce large amounts of proinflammatory cytokines such as IFN-γ and TNF-α and also have cytotoxic potential, which may cause tissue damage and development of pathogenesis in many inflammatory disorders. Here we review the characteristics of CD4⁺CD28⁻ T cells as well as the recent advances highlighting the contribution of these cells to several disease conditions.
Subject(s)
Humans , Cellular Senescence , Cytokines , Memory , T-LymphocytesABSTRACT
Objective: To study the mechanism of attenuation by stir baking with vinegar on effect of ethyl acetate fraction of Kansui Radix on mice gastrointestinal permeability. Methods: Mice were randomly divided into blank control, Kansui Radix, and Kansui Radix stir-baked with vinegar groups. The gastric and intestinal tissues were removed from mice after ig administration for 7 d and were observed by the transmission electron microscope (TEM). The expression of E-cadherin and VCAM-l was examined by immunohistochemical techniques. Results: Using TEM, the arrangement and karyon morphology of gastric glandular epithelium cells were irregular with the uneven electronic of cytoplasm and lots of lysosomes in Kansui Radix group; The morphology of the gastric epithelial cells was irregular with some abnormal mitochondrions in the cytoplasm in Kansui Radix stir-baked with vinegar group; The intestinal mucosa cells were covered with short microvilli with well-connected intercellular junctions, but the tight junctions on the near-surface were short in Kansui Radix group; The arrangement and morphology of intestinal epithelial cells were fairly uniform with few and short microvilli and intact mitochondrion in Kansui Radix stir-baked with vinegar group. Using immunohistochemical techniques, compared with the control group, Kansui Radix could decrease the expression of E-cadherin and increase the expression of VCAM-l (P < 0.05, 0.01). While compared with Kansui Radix group, Kansui Radix stir-baked with vinegar could obviously increase the expression of E-cadherin (P < 0.05, 0.01) and decrease the expression of VCAM-l (P < 0.05, 0.01). Conclusion: Stir-baking with vinegar can attenuate the Kansui Radix-induced gastrointestinal inflammation. The mechanism may be that it can improve the morphology of mice gastrointestinal mucosal cells, reduce the membrane permeability, and decrease the stimulating effect of Kansui Radix.