Suppressor of cytokine signaling 1 expression during LPS-induced inflammation and bone loss in rats

Abstract This study aimed to characterize the dynamics of suppressor of cytokine signaling (SOCS1) expression in a rat model of lipopolysaccharide-induced periodontitis. Wistar rats in the experimental groups were injected three times/week with LPS from Escherichia coli on the palatal aspect of the first molars, and control animals were injected with vehicle (phosphate-buffered saline). Animals were sacrificed 7, 15, and 30 days after the first injection to analyze inflammation (stereometric analysis), bone loss (macroscopic analysis), gene expression (qRT-PCR), and protein expression/activation (Western blotting). The severity of inflammation and bone loss associated with LPS-induced periodontitis increased from day 7 to day 15, and it was sustained through day 30. Significant (p < 0.05) increases in SOCS1, RANKL, OPG, and IFN-γ gene expression were observed in the experimental group versus the control group at day 15. SOCS1 protein expression and STAT1 and NF-κB activation were increased throughout the 30-day experimental period. Gingival tissues affected by experimental periodontitis express SOCS1, indicating that this protein may potentially downregulate signaling events involved in inflammatory reactions and bone loss and thus may play a relevant role in the development and progression of periodontal disease.

Expression and Clinical Significance of TRIM55 in Colorectal Cancer / 胃肠病学

TRIM55 is a member of TRIM family.Most TRIM proteins, which can be defined as E3 ubiquitin ligase because of the RING-finger domain, are closely related with the initiation and progression of cancer.Aims: To study the expression and clinical significance of TRIM55 in colorectal cancer, and explore the potential mechanism of TRIM55 in colorectal cancer.Methods: Seventy colorectal cancer tissues and corresponding paracancerous tissues taken from colorectal cancer patients from October 2014 to December 2015 at Shanghai Ren Ji Hospital were enrolled.Real-time PCR was performed to examine the expression of TRIM55.Human colorectal cancer cell line HCT116 was transfected with TRIM55 small interfering RNA (siRNA), cell proliferation was measured by CCK-8 assay, Western blotting was implemented to determine the protein expressions of TRIM55 and SOCS1.Results: The expression of TRIM55 was significantly increased in 49 colorectal cancer tissues than in corresponding paracancerous tissues.Increased expression of TRIM55 was closely correlated with tumor differentiation (P=0.032), AJCC staging (P=0.001) and lymph node metastasis (P=0.001), but not related to gender, age, tumor size, invasion depth, distant metastasis and vascular invasion (P>0.05).After transfection with TRIM55 siRNA, mRNA and protein expressions of TRIM55 were significantly decreased (P<0.01), proliferation of HCT116 cells was significantly decreased (P<0.01), and protein expression of SOCS1 was significantly increased (P<0.01).Conclusions: E3 ubiquitin ligase TRIM55 may promote the proliferation of colorectal cancer cells via influencing the expression of SOCS1, thus promoting the progression of colorectal cancer.This indicates that detection of TRIM55 expression may provide a new approach for diagnosis and therapy of colorectal cancer.

The effects and mechanism of SOCS3 gene transfection in CD4+Th cell differentiation and expression of inflammatory cytokines of mouse / 重庆医学

Objective To investigate the effect and mechanism of adenovirus vector mediated SOCS 3 gene transfection in CD4+ Th cell differentiation and expression of inflammatory cytokines of mouse .Methods The CD4+ Th cells were isolated from spleen of C57bl/6 mouse and cultured .Ad‐SOCS3 were transfected into the CD4+ Th cells .PHA was used for culturing with the CD4+ Th cells .RT‐PCR were used to detect the mRNA expression ,and Western blot were used to detect the protein expression of cytokines .Results Compared with the control group ,the gene and protein expression of T‐bet ,IL‐2 ,IFN‐γ,STAT4 and IL‐12Rβ2 in the transfected group were significantly down‐regulated ,the gene and protein expression of SOCS3 ,GATA‐3 ,IL‐4 ,IL‐6 ,IL‐10 and STAT6 were significantly up‐regulated(P<0 .01) .Conclusion The results indicate that SOCS3 gene transfection can up‐regu‐late SOCS3 mRNA and protein expression in the CD4+ Th cells ,down‐regulate the JAK/STAT pathway ,inhibition of Th1 cell dif‐ferentiation ,and down regulation of inflammatory cytokine gene and protein expression ,and indirectly promote Th2 cell differentia‐tion ,and up the corresponding inflammatory cytokine gene and protein expression .

Effect of gypenoside on lipopolysaccharide-mediated microglial inflammatory response / 国际脑血管病杂志

Objective To investigate the effect of gypenoside on lipopolysaccharide (LPS)-mediated inflammatory response. Methods The BV2 microglia cell line was cultured in vitro. The BV2 microglia cells were divided into four groups: normal control, LPS (10 ng/ml), GP + LPS (GP 20 μg/ml, LPS 10 ng/ml), and GP (20 μg/ml). After 24 h cultivation, ELISA was used to detect the levels of tumor necrosis factor α(TNF-α), interleukin (IL)-1β, and IL-6. Immunocytochemistry staining and Western blot were used to detect the expression levels of nuclear factor (NF-κB) and suppressor of cytokine signaling 1 (SOCS-1). Results Compared with the normal control group, the release of TNF-α, IL-1β and IL-6, as well as the expression level of NF-κB in the LPS group were increased significantly (all P 0. 05 ). Conclusions GP can significantly inhibit the LPS-mediated microglial inflammatory response. SOCS-1 protein may be involved in GP inhibiting LPS-mediated microglial inflammatory response.

Avaliação de polimorfismos genéticos como biomarcadores na evolução da cardiomiopatia chagásica / Evaluation of genetic polymorphisms as biomarkers in the evolution of Chagas cardiomyopathy

O Trypanosoma cruzi é um parasita intracelular e agente causador da doença de Chagas, que afeta milhões de pessoas em todo o mundo. Sabe-se que durante os processos de inflamação, regeneração e fibrose desencadeados pelo T. cruzi no hospedeiro há a participação de diversos mediadores e fatores. O objetivo deste trabalho foi avaliar a associação entre polimorfismos de nucleotídeos únicos com as formas clínicas e o grau de fibrose em pacientes com doença de Chagas. Os polimorfismos foram analisados por PCR em tempo real. Foram incluídos no estudo 55 pacientes com diagnóstico de doença de Chagas e classificados de acordo com a forma clínica da doença, sendo que 17 apresentavam a forma indeterminada, 15 a forma cardíaca sem disfunção ventricular e 23 a forma cardíaca com disfunção ventricular. Os genótipos CA dos polimorfismos do gene LGALS3 (rs4644 e rs4652); AG e GG do SOCS3 (rs4969170); CT e TT do IL-28B (rs12979860 e 8099917, respectivamente); AG, AG, CC, AG e AG do CLDN-1 (rs10212165, rs3909582, rs9865082, rs9880018 e rs9848283, respectivamente); e CC do CCL5 (rs2280789) foram estatisticamente mais frequentes em pacientes com a forma cardíaca do que com a forma indeterminada da doença. Com relação ao grau de fibrose, os genótipos CC dos polimorfismos do gene LGALS3 (rs4644 e rs4652); AA do SOCS3 (rs4969170); CC do rs12979860 e TT do rs8099917 do IL-28B; AA do rs10212165, AA, AG e GG do rs3909582, CC e CT do rs9865082, AG e GG do rs9880018 e AA do rs9848283 do gene CLDN1; e CC do CCL5 (rs2280789) foram estatisticamente mais frequentes em indivíduos com fibrose cardíaca <15% quando comparados com o grupo com fibrose ≥15%. Diante do exposto concluimos que os polimorfismos analisados podem ser úteis como futuros biomarcadores para estadiamento e conduta terapêutica em pacientes com doença de Chagas.

Trypanosoma cruzi is an intracellular parasite and the agent that causes Chagas disease, which affects millions of people worldwide. Several factors and mediators are known to actively participate in the inflammation, fibrosis and tissue regeneration, which is triggered by T. cruzi within the host. The aim of this study was to evaluate the association of single nucleotide polymorphisms with clinical forms and rate of fibrosis in Chagas disease patients. The polymorphisms were analyzed by real-time PCR. The study consisted of 55 Chagas disease patients that were classified according to the clinical form of the disease, including 17 patients presenting the indeterminate form, 15 patients presenting the cardiac form without ventricular dysfunction and 23 patients presenting the cardiac form with ventricular dysfunction. The genotypes of CA of LGALS3 gene polymorphisms (rs4644 and rs4652); AG and GG of SOCS3 (rs4969170); CT and TT of IL-28B (rs12979860 and 8099917, respectively); AG, AG, CC, AG and AG of CLDN-1 (rs10212165, rs3909582, rs9865082, rs9880018 and rs9848283, respectively); and CC of CCL5 (rs2280789) were significantly more frequent in patients presenting the cardiac form compared to patients presenting the indeterminate form. Regarding the degree of fibrosis, the CC genotype of polymorphisms of the genes LGALS3 (rs4644 and rs4652); AA of SOCS3 (rs4969170); CC of rs12979860 and TT of rs8099917 of the IL-28B; AA of rs10212165 and AA, AG and GG of rs3909582, CC and CT of rs9865082, AG and GG of rs9880018 and AA of rs9848283 of the gene CLDN1; and CC of CCL5 (rs2280789) were statistically more frequent in patients presenting <15% cardiac fibrosis when compared to patients presenting fibrosis ≥15%. Taken together, our results suggest that the polymorphisms analyzed may be useful biomarkers for therapeutic management of patients with Chagas disease.

Effects of Suppressor of Cytokine Signaling-1 Silencing on the Proliferation and Interferon Gamma Susceptibility of Human Pancreatic Cancer Cells / 天津医药

Objective To detect the changes of cell proliferation and IFN-γsusceptibility of human pancreatic can-cer cells after suppressor of cytokine signaling-1 (SOCS1) gene silencing, and to explore the SOCS1 as the target of anti-tu-mor therapy through enhancing the function of IFN-γ. Methods Western blot assay, PCR and real-time PCR were used to verify the down regulation of SOCS1 in human pancreatic cancer cell (PANC1) after transfection;subsequently, PANC1 was stimulated with IFN-γ. Western blot assay was also used to detect the expression of signal transducers and activators of tran-scription (STAT)1 and phosphorylation STAT(pSTAT)1;and the change of IFN-γsusceptibility was detected by MTT assay. Real-time PCR was used to detect the mRNA of interferon regulatory factor-1(IRF-1). Flow cytometry was used to detect the cell cycle. Results The expression levels of SOCS1 mRNA and protein were significantly decreased in small hairpin SCOS1 (shSOCS1) transfected PANC1 cells. After the silence of SOCS1, the expression levels of IRF-1 and pSTAT1 in-creased significantly (P<0.05), and the median inhibitory concentration(IC50)of IFN-γfor PANC1 cells decreased signifi-cantly (P<0.01). The cell count of shSOCS1 cells dropped significantly compared with that of control group after the SOCS1 silencing for 72 hours (P<0.05). The cell cycle arrest was promoted at the G0/G1 phase, but the percentage of cells in S phase and G2/M decreased compared to that of control groups (P<0.05). Conclusion After the inhibition of SOCS1 gene expression, the proliferation ability of human pancreatic cancer cell line PANC1 decreased, and the sensitivity of PANC1 cells to IFN-γwas enhanced.

Suppressor of cytokine signaling-3 improves proliferation and migration of human trophoblast cells during pre-eclampsia / 中华围产医学杂志

Objective To investigate the expression of suppressor of cytokine signaling-3 (SOCS-3) gene in placenta,its role in the pathogenesis of pre-eclampsia and its effect on proliferation and migration of HTR-8/SVneo cells.Methods Fifteen women with severe pre-eclampsia hospitalized in the First Affiliated Hospital of Nanjing Medical University from October 2010 to March 2011 and t 5 normal pregnant women during the same time period were investigated.Cultured HTR-8/SVneo cells were transfected with SOCS-3 specific small interfering RNA (siRNA) or negative siRNA as the controls.The expression of SOCS-3 mRNA and protein in placenta and these cells was detected by real-time quantitative reverse transcription-polymerase chain reaction and Western blot.Cell proliferation was detected by methyl thiazolyl tetrazolium,cell cycle by flow cytometry and migration by the Transwell test.Two independent t tests were used for statistical analysis.Results The SOCS-3 mRNA and protein levels in the severe pre-eclampsia group were lower than those in the normal group (0.25±0.03 vs 0.71±0.08 and 0.21±0.05 vs 0.75±0.12,t=15.94 and 14.29,respectively,both P＜0.05).SOCS-3 mRNA and protein levels in the transfection group at 24 hours were lower than those in the negative control group (0.39±0.02 vs 1.00±0.04 and 0.003 7±0.001 4 vs 1.514 9±0.035 7,t=27.58 and 73.35,respectively,both P＜0.05).The integral absorbance values of cell proliferation in the transfection group at 48,72 and 96 hours after transfection were 0.23 ± 0.01,0.32±0.02 and 0.37± 0.02,respectively,which were lower than those in the negative control group (0.39± 0.02,0.55 ± 0.04 and 0.86± 0.04,t=2.60,6.64 and 42.44,respectively,all P＜0.05).The cell clonal formation was lower in the transfection group compared with the negative group (116± 15 vs 312±24,t=9.96,P＜0.05).The ratios of G1/G0 and S phase cells in the transfection group were (55.75±2.21) ％ and (31.59±0.83) ％,respectively,and were significantly different from those in the negative control group [(47.88± 1.87) ％ and (37.38± 1.34) ％,t=45.43 and 20.06,respectively,P＜0.05].After 48 hours,cell migration in the transfection group was lower than that in the negative control group (93 ± 11 vs 167± 17,t=21.36,P＜O.05).Conclusion SOCS-3 expression is probably involved in the pathogenesis of pre-eclampsia by being down-regulated and therefore impeding proliferation and migration of the trophoblast.

Mechanisms of STAT family mediated IDO regulation in dendritic cells / 国际肿瘤学杂志

Indoleamine 2,3-dioxygenase (IDO) is known as an endogenous immunosuppressive enzyme which plays a significant role in the process of tumor.IDO is not only found in tumor cells but also detected in dendritic cell (DC) in tumor microenvironment,which participates in the formation of tumor immune tolerance through expressing IDO enzyme.Signal transducer and activator of transcription (STAT) is the main signal protein family which participates in the IDO transcriptional regulation of DC.It is necessary to detail the signaling pathway in regulating IDO expression,which will help us develop high specific and more active IDO inhibitors and provide new options for anti-cancer targeted therapy.

Expression of SOCS1 and SOCS3 genes in human graft-versus-host disease after allogeneic hematopoietic stem cell transplantation

BACKGROUND: Suppressor of cytokine signaling genes (SOCS) are regarded as pivotal negative feedback regulators of cytokine signals, including the interferon-gamma (IFN-gamma), granulocyte-colony stimulating factor, and interleukin families, released by T cells. A detailed understanding of the involvement of SOCS genes in graft-versus-host disease (GVHD) is critical to effectively manage GVHD, yet their expression patterns among recipients remain largely unexplored. METHODS: Expression levels of SOCS1 and SOCS3 were determined by real-time quantitative reverse transcription PCR (qRT-PCR) in patients with acute GVHD (aGVHD) and chronic GVHD (cGVHD), in a severity-dependent manner, after allogeneic hematopoietic stem cell transplantation (HSCT). A total of 71 recipients with AML (N=40), ALL (N=12), myelodysplastic syndromes (MDS; N=10), chronic myelogenous leukemia (CML; N=2), severe aplastic anemia (SAA; N=5), or others (N=2), who received allogeneic HSCT from human leukocyte antigen-identical siblings or unrelated donors between 2009 and 2011, were included in the present study. RESULTS: Overall, the expression levels of SOCS1 decreased in recipients with grade II to IV aGVHD and cGVHD when compared to normal donors and non-GVHD recipients. Interestingly, the expressions of SOCS1 decreased significantly more in cGVHD than in aGVHD recipients (P=0.0091). In contrast, SOCS3 expressions were similarly reduced in all the recipients. CONCLUSION: This is the first study to show that SOCS1 and SOCS3 are differentially expressed in recipients following allogeneic HSCT, suggesting a prognostic correlation between SOCS genes and the development of GVHD. This result provides a new platform to study GVHD immunobiology and potential diagnostic and therapeutic targets for GVHD.

Expression of SOCS1 and SOCS3 genes in human graft-versus-host disease after allogeneic hematopoietic stem cell transplantation

BACKGROUND: Suppressor of cytokine signaling genes (SOCS) are regarded as pivotal negative feedback regulators of cytokine signals, including the interferon-gamma (IFN-gamma), granulocyte-colony stimulating factor, and interleukin families, released by T cells. A detailed understanding of the involvement of SOCS genes in graft-versus-host disease (GVHD) is critical to effectively manage GVHD, yet their expression patterns among recipients remain largely unexplored. METHODS: Expression levels of SOCS1 and SOCS3 were determined by real-time quantitative reverse transcription PCR (qRT-PCR) in patients with acute GVHD (aGVHD) and chronic GVHD (cGVHD), in a severity-dependent manner, after allogeneic hematopoietic stem cell transplantation (HSCT). A total of 71 recipients with AML (N=40), ALL (N=12), myelodysplastic syndromes (MDS; N=10), chronic myelogenous leukemia (CML; N=2), severe aplastic anemia (SAA; N=5), or others (N=2), who received allogeneic HSCT from human leukocyte antigen-identical siblings or unrelated donors between 2009 and 2011, were included in the present study. RESULTS: Overall, the expression levels of SOCS1 decreased in recipients with grade II to IV aGVHD and cGVHD when compared to normal donors and non-GVHD recipients. Interestingly, the expressions of SOCS1 decreased significantly more in cGVHD than in aGVHD recipients (P=0.0091). In contrast, SOCS3 expressions were similarly reduced in all the recipients. CONCLUSION: This is the first study to show that SOCS1 and SOCS3 are differentially expressed in recipients following allogeneic HSCT, suggesting a prognostic correlation between SOCS genes and the development of GVHD. This result provides a new platform to study GVHD immunobiology and potential diagnostic and therapeutic targets for GVHD.

Expression and significance of SOCS3 in liver tissue of rats with severe acute pancreatitis complicated by liver injury / 解放军医学杂志

Objective To investigate the expression and mechanism of action of suppressor of cytokine signaling 3 (SOCS3) in liver tissue of rats with experimental severe acute pancreatitis (SAP) concurring with liver injury. Methods The rat model of SAP was reproduced by retrograde injection of 4% sodium taurocholate into the biliopancreatic duct. Thirty-two male SD rats were randomly assigned into 4 groups (8 each): normal control group (NC), SAP 6h, 12h, and 18h groups. The levels of serum amylase (AMY), alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were measured dynamically. The concentrations of IL-6 and IL-18 were determined by ELISA. The localization and expression of SOCS3 protein in liver were determined by immunohistochemical staining and Western blotting. Results Compared with NC group, the serum levels of AMY, ALT and AST increased significantly in SAP groups (P<0.05), and there was significant difference among SAP groups. The serum concentrations of IL-6 and IL-18 increased significantly in the SAP groups than in NC group (P<0.05), and there was significant difference among SAP groups. Compared with NC group, the concentration of SOCS3 protein increased significantly in SAP groups, and increased gradually along with the increased duration of pancreatitis (P<0.05). A minor expression of SOCS3 protein was found in NC group. The change in SOCS3 protein concentration was consistent with the severity of liver injury as well as the serum concentrations of IL-6 and IL-18. Conclusions The inflammatory action induced by SAP concurring with liver injury may induce the expression of SOCS3 in liver tissue, and it may increase in intensity along with the severity of liver injury and inflammatory reaction. The mechanism may be attributed to a negative feedback regulation of the inflammatory action mediated by JAK/STAT pathway.

Expression of cytokines and suppressor of cytokine signaling in peripheral blood mononuclear cells derived dendritic cells from primary biliary cirrhosis patients / 中华检验医学杂志

Objective To study the role of suppressor of cytokine signaling ( SOCS ) in the pathogenesis of primary biliary cirrhosis( PBC),the levels of SOCS protein and the changes of function of dendritic cell(DC) were respectively observed from PBC patients.MethodsThe study population consisted of 10 patients of PBC and 8 healthy controls.Phenotypic analysis of cultured peripheral blood mononuclear cells (PBMC)-derived DC was performed by flow cytometry (FCM),such as CD83,CD86 and human leukocyte antigen DR (HLA-DR).The levels of interleukin-10 (IL-10),interferon-γ( IFN-γ) and IL-12 in culture supematant of DC were measured by enzyme linked immunosorbent assay ( ELISA ).The protein levels of SOCS1 and SOCS3 were detected by Western blot ( WB ).The features of changes in these parameters were analyzed between the two groups.ResultsThe expression of CD83,CD86 and HLA-DR in PBC patients were ( 79.4 ± 4.8 ) ％,( 86.5 ± 6.3 ) ％ and (90.0 ± 3.5 ) ％,which were significantly higher than those in healthy control group[ (68.3 ±4.1 )％,(74.2 ±6.3)％ and (83.6 ±7.6)％ ],respectively (t =5.340,4.120,2.514,P ＜0.05).The levels of IL-12 and IFN-γin PBC patients were (53.5 ± 11.1)and (32.0 ±9.0) ng/L,which were significantly higher than those in healthy control group[ (32.1 ± 10.7) and (15.4 ± 8.1 ) ng/L; t =4.123,3.818,P ＜ 0.01 ].There were not any significant difference of IL-10 level between PBC patients [ (7.0 ± 4.6) ng/L ] and the healthy controls [ ( 5.8 ± 4.2) ng/L; t =0.563,P ＞ 0.05 ].The proteins levels of SOCS1 and SOCS3 in PBMC-derived DC from PBC group were decreased significantly than those in healthy control group.ConclusionsThe results suggest that the PBMC-derived DC in PBC patients has greater ability of potent maturation and antigen presentation function.The decreased expression of SOCS levels may be associated with the excessive immunological reaction and the breakdown of self-tolerance.

Effects of β-elemene combined with chemotherapy on the protein expressions in lung adenocarcinoma A549 cell line / 中华老年医学杂志

ObjectiveTo explore the possible molecular mechanisms of β-elemene combined with cisplatin and heat therapy for killing A549 cell line.MethodsThe protein expressions of Stat3,p21 Waf1/Cip1 and Survivin were detected with Western blot after treatment with β-elemene of different concentrations combined cisplatin and heat therapy.ResultsThe protein expressions of Stat3,and pStat3 and p21Waf1/Cip1 in A549 cell line were enhanced with increasing concentrations,and there was significant difference in the expressions between high and low concentration,but Survivin protein had no change at 37°C.After adding 4 μg/mL cisplatin,the expressions of p21Waf1/ Cip1,Survivin,Stats and pStat3 were reduced at β-elemene of high concentration.At 42°C,there was no significant difference in expression of Stat3 protein at 60 μg/mL elemene,but the expressions of pStat3,p21Waf1/Cip1 and Survivin proteins had sharply declined.When using 15 μg/mL elemene combined with 4 μg/mL cisplatin,the protein expressions of Star3 and pStat3 increased,and Survivin expression decreased.ConclusionsAt temperature of 37°C,β-elemene of high concentration may inhibit growth of A549 cells by higher expression of p21Waf1/Cip1 protein,and mainly by inhibiting expressions of Stat3 and pStat3 and Survivin after combined with cisplatin.At temperature of 42°C,β- elemene of highconcentrationmaypromoteapoptosispossiblythroughinhibition ofStat3 phosphorylation and expression of Survivin protein.β-elemene of low concentration combined with cisplatin leads to synergy killing effect by reducing expression of Survivin protein.

Expression of suppressor of cytokine signaling 3 and its significance in human placenta with pregnant intrahepatic cholestasis / 中华妇产科杂志

Objective To detect the expression and significance of suppressor of cytokine signaling 3(SOCS3) in the placentas of pregnant women with intrahepatic cholestasis of pregnancy (ICP). Methods Totally, 44 ICP gravidas, including 21 severe ICP and 23 mild ICP who delvered through cesarean section at the Second Xiangya Hospital of Central South University from April 2008 to January 2009, were selected as the ICP group, and another 25 healthy pregnant women were chosen as control. Placentas of the above gravidas were collected and the expression and localization of SOCS3 were determined by immunohistochemical peroxidase streptomyces-avidin link (SP) method (indicated by the percentage ofpositive cells and average gray scale) and the levels of interleukin-10 (IL-10) and interferon-γ (IFN-γ)from placenta homogenation were measured by enzyme linked immunoadsorbent assay (ELISA). Results(1) SOCS3 were expressed in placentas of both groups mainly in the intracytoplasma of trophocyte.However, weakly positive, positive, and strongly positive expressions were found in the severe ICP, mild ICP and the control group, respectively. Almost no expression was detected in membrane and nucleus of the trophoblasts. (2) The percentage of SOCS3 positive cells in the severe ICP group was significantly lower than in the control and the mild ICP group, respectively [ (0.15±0.08)% vs (0.69±0.12)% and (0.42±0.09) % , P < 0.01 ]. The average gay SOCS3 in placental tissue in the severe ICP group was significantly higher than that in control and mild ICP group, respectively (204 ±7 vs 81 ±7 and 147 ± 7, P <0.01 ).(3) Significant lower level of IL-10 in placenta homogenation was found in the severe ICP group than in the control and mild ICP group [ ( 1.16 ± 0.68) μg/L, vs ( 1.39 ± 0.08) μg/L and ( 1.22 ± 0.75 ) μg/L,P < 0.01 ]. (4) The opposite results were found in the level of IFN-γin trophoblasts and the ratio of IFN-γ/IL-10 [ severe ICP group: ( 16.8 ± 0.7 ) μg/L and 16.02 ± 2.79; control group: ( 10.5 ± 0.3 ) μg/L and8.56 ±0.14; mild ICP group: ( 13.4 ±0.5) μg/L and 8.56 ±0.14, P <0.01]. (5) Negative correlation was shown between the percentage of SOCS3 positive cells in trophoblasts and the ratio of IFN-γ/IL-10 ( r =-0.685 and -0.702, P < 0.01 ), and the average gay SOCS3 was positively correlated with the ratio of IFN-γ/IL-10 (r=0.621 and 0.891, P<0.01) in both mild and severe ICP group. Conclousions SOCS3 may participate in the pathogenesis of ICP and its expression may affected by the severity of ICP, and SOCS3may also play a role in the immunological regulation in ICP patients.

Suppressor of cytokine signaling-1 inhibits high glucose-induced expression of monocyte chemoattractant protein-1 in glomerular mesangial cells / 中华肾脏病杂志

Objective To investigate the effect of suppressor of cytokine signaling-1 (SOCS-1)on expression of monocyte chemoattractant protein-1 (MCP-1)in human glomerular mesangial cells (HMCs) under high concentration of glucose. Methods Stable transfections of HMC with pCR3.1 vector and pCR3. 1-SOCS-1 were performed with hpofectamine 2000, and cells were selected with geneticin. Cells were stimulated with low glucose (LG, 5.5 mmol/L), high glucose (HG, 30 mmol/L), LG plus mannitol (24.5 mmol/L) and AG490 (10 μmol/L). The protein expression levels of SOCS-1, signal transducer and activators of transcription 1,3 (STAT1, STAT3),p-STAT1 and p-STAT3 were observed by Western blotting. The protein synthesis of MCP-1, FN and type Ⅳ collagen in the supernatants of the HMCs were detected by ELISA and radioimmunoassay. The expression level of SOCS-1 and MCP-1 mRNA was measured by BT-PCR.Results HG induced the expression of SOCS-1 protein and mRNA in HMCs in time-dependant manner, peaked at 4 h, and gradually decreased to baseline at 24 h. Compared with low glucose control group, the phosphorylation levels of STAT1 and STAT3 and the expression of MCP-1 mRNA [(0.39±0.05) vs (0.16±0.02)]were significantly increased in HMCs under high glucose medium (P ＜0.01 ). Exposure of HMCs to high glucose conditions showed high concentration of MCP-1 [(459±67) ng/L vs (241±19) ng/L], fibronectin [(5.84±0.61) mg/L vs (3.41±0.31) mg/L]and type Ⅳ collagen [(16.45±2.30) μg/L vs (9.56±1.52) μg/L]in the supernatants (all P＜0.01).Overexpression of SOCS-1 inhibited the phosphorylation levels of STAT1 and STAT3 and the expression of MCP-1 mRNA [(0.34±0.04) vs (0.42±0.05)]in HMCs under high glucose condition (all P＜0.05). Compared with vector control group, the concentration of MCP-1 [(387±47) ng/L vs (463±56) ng/L], fibronectin [(4.61±0.57) mg/L vs (5.76±0.74) mg/L]and type Ⅳ collagen [(13.4±2.32) μg/L vs (17.1±2.57) μg/L]was decreased in supernatants of HMCs with SOCS-1 overexpression (all P＜0.05). Compared with HG group, the expression of MCP-1 mRNA (0.31±0.04) and the concentration of MCP-1 [(361±53) ng/L], FN [(5.46±0.71) mg/L]and type Ⅳ collagen [(15.2±1.97) μg/L]in supernatants were decreased in HMCs treated with AG490.Conclusion Overexpression of SOCS-1 inhibits overproduction of MCP-1 and ECM proteins in HMCs under high glucose conditions, which may be partly by regulating the phosphoryhtion of STAT1 and STAT3.

Effect of Acupuncture-moxibustion on the Expression of IGF-1 and SOCS2 in Colonic Mucosa of Rats with Ulcerative Colitis / 针灸推拿医学（英文版）

Objective: To observe the effect of acupuncture-moxibustion on the expression of insulin-like growth factor-1 (IGF-1) and suppressor of cytokine signaling-2 (SOCS2) in colonic mucosa of rat models of ulcerative colitis (UC), and explore the mechanism of acupuncture- moxibustion therapy in treating UC. Methods: The rats were randomized into a normal control (NC) group, a model control (MC) group, an herb-partitioned moxibustion (HPM) group and an electroacupuncture (EA) group, 8 in each group. The rat models of UC were established by immunological methods combined with local stimulation. The rats in the HPM and EA groups were given herb-partitioned moxibustion and electroacupuncture treatments respectively, once every day, lasting for 14 d. The morphological variations of rat's colonic mucosa were observed under light microscope; the colonic mucosal mucin was detected by PAS-AB and HID-AB staining methods; the expression of IGF-1 and SOCS2 was assayed by the immunohistochemical method. Results: In the rat models of UC, ulceration and inflammation of the colon were revealed by light microscope. The concentration of colonic mucosal mucin was reduced (P<0.01), while the expression of IGF-1 had an increase (P<0.01), and the expression of SOCS2 was reduced (P<0.01). After HPM or EA treatment, the pathological injuries of colonic mucosa had improved, the concentration of mucin increased (P<0.01), the expression of IGF-1 decreased (P<0.01), and the expression of SOCS2 increased (P<0.01). Conclusion: The secretion of mucosal mucin in rat UC decreased, the expression of IGF-1 was significantly higher, while the expression of SOCS2 was remarkably lower; both HPM and EA can help improve the damage of colonic mucosa in rat UC, and modulate the secretion of mucin, as well as regulate the expression of IGF-1 and SOCS2 in the colonic mucosa.

Study on the aberrant methylation of SOCS-1 gene in diffuse large B-cell lymphoma / 白血病·淋巴瘤

Objective To explore the significance of the suppressor of cytokine signaling-1 (SOCS-1)gene methylation in the genesis, development and prognosis of diffuse large B-cell lymphoma (DLBCL).Methods The methylation state of CpG island in SOCS-1 gene were detected by methylation-specific polymerase chain reaction (PCR) and the level of SOCS-1 gene was measured by the real-time PCR. The clinical data of 30 patients with DLBCL were collected, and according to the international prognosis index (IPI), they were divided into low risk group and high risk group. Results Aberrant methylation of SOCS-1 in 17 DLBCL patients (56.7 %) were positive, however, in control group aberrant methylation was negative(P ＜0.01).The methylation level of DLBCL patients with positive SOCS-1 methylation was higher than that of patients with negative (P ＜0.05). Combined with the clinical data, the positive rate of methylation in patients with high level of serum LDH or the numbers of extra-nodal lesions＞l were significantly higher than that in patients with normal LDH level or the numbers of extra-nodal lesions ≤ 1, respectively. Hence, the positive rate of methylation in the high risk group of DLBCL was higher than that in the low risk group (P ＜0.05). Conclusion There were aberrant methylations of the SOCS-1 gene in the patients with DLBCL. The methylations of SOCS-1 can silence the gene expression, which indicates that SOCS-1 and its methylations play some role on genesis and development of DLBCL and can evaluate the prognosis of the patients with DLBCL.

Effects of triptolide on interferon-γ signaling in a human keratinocyte cell line HaCaT / 中华皮肤科杂志

Objective To investigate the effects of triptolide on the expression of a series of proteins associated with interferon-γ (IFN-γ)signaling in HaCaT keratinocytes.Methods After pretreatment with difrerent dosages of triptolide(10-10-10-7 mol/L),HaCaT cells were stimulated by recombinant human IFN-γ(rhIFN-γ,500 U/mL)for various periods followed by the collection of cells.Then,total protein was extracted from these cells and subjected to Western blotting for the detection of expression of interferon-γ receptor α(IFN-γRα),phosphorylated Janus kinase 2(pJAK2)and suppressor of cytokine signaling (SOCS1).Results Triptolide at the concentrations of 10-8 mol/L and 10-7 mol/L significantly inhibited the IFN-γRα expression upregulated by rhIFN-γ(both P＜0.05).The expression of pJAK2 induced bv rhIFN-γ was also suppressed by triptolide at the concentrations of 10-9 moI/L and 10-8 mol/L(both P＜0.05).The inhibition of triptolide on IFN-γRα and pJAK2 expression was dose-dependent and the 50%inhibitory concentrations(IC50 value)were 1.37×10-8 mol/L and 2.83×10-9 mol/L,respectively.On the contrary,triptolide upregulated the expression of SOCS1 stimulated by rhIFN-γ at the concentrations of 10-10,10-9 and 10-8 mol/L(P＜0.05,0.05,0.01,respectively)with the 50%effective dosage(ED50 value)at 3.32 × 10-11 mol/L.Conclusions By inhibiting the expression of IFN-γRα as well as phosphorylation of JAK2 and upregulating the expression of SOCS1,triptolide inhibits the phosphorylation of STAT-1,resulting in the inhibition of genetic transcription of multiple inflammatory factors induced by IFN-γ signaling in HaCaT keratinocytes,and the inhibition probably contributes to the efficacy of triptolide in the treatment of IFN-γ-dependent inflammatory skin disorders,such as psoriasis.

Expressions and relationship of suppressor of cytokine signaling1 and c-myc in human hepatocellular carcinoma tissue / 中国现代普通外科进展

Objective:To study the expression of suppressor of cytokine signaling 1(SOCS1) and cellular myelocytomatosis oncogene(c-myc)in hepatocellular carcinoma(HCC),and to inves-tigate the relation between the expression of SOCS1 and c-myc in HCC.Methods:The expressions of SOCS1 and c-myc protein of 41 HCC tissues and surrounding tissues,as well as the tissues of 11 normal cases,were detected by immunohistochemical(IHC)staining using SP method.The expression of SOCS1 mRNA was detected by RT-PCR.And the clinical and pathological data were analysed.Results:Compared with the para-carcinoma and normal liver tissue,the levels of SOCS1 and SOCS1 mRNA in HCC tissues were significantly lower(P

Expressions of signal transducer and activators of transcription 3 (STAT3) and suppressor of cytokine signaling 3 (SOCS3) proteins in breast carcinoma and their association with biological behaviors / 肿瘤

Objective: To investigate the expressions of signal transducer and activators of transcription 3 (STAT3) and suppressor of cytokine signaling 3 (SOCS3) protein in the breast carcinoma tissue and their relationship with tumor differentiation, invasion, and metastasis. Methods: The expression of STAT3, phospho-STAT3 and SOCS3 protein were determined by tissue microarray and immunohistochemistry with EnVision system in 71 cases of archival breast carcinoma tissues and 41 cases of noncancerous tissues. The relationship between their expression and the clinical pathological parameters were analyzed. Results: (1) The positive rates of STAT3, phospho-STAT3, and SOCS3 was 78.9%, 69.0%, and 29.6%, respectively. The difference was significantly different compared with control (P0.05). The expression of SOCS3 was negatively related with the histopathologic grade and the axillary lymph node metastasis (P0.05). (3) The expression of STAT3 and phospho-STAT3 had negative correlation with the expression of SOCS3 in breast carcinoma tissues (P<0.01). Conclusion: The overexpressions of STAT3 and phospho-STAT3 and the down-regulated expression of SOCS3 closely correlated with the tumor carcinogenesis, progression, invasion, and metastasis of breast carcinoma. Detection of their expression is helpful in accessing the malignant degree and the biological behaviors of breast carcinoma.