Curcumin Induces Cycle Arrest of Colon Cancer HCT116 Cells via JAK1/STAT1/p21 Pathway / 中国实验方剂学杂志

ObjectiveTo investigate the effect of curcumin on the cycle arrest of human colon cancer HCT116 cells and decipher the possible molecular mechanism. MethodThe methyl thiazolyl tetrazolium （MTT） method was employed to examine the effects of curcumin （0， 12.5， 25， 50， 75， 100 μmol·L-1） and 5-fluorouracil （5-FU， 600 μmol·L-1） on the proliferation of HCT116 cells at different time points （24， 48， 72 h）. Flow cytometry was employed to examine the cycle of HCT116 cells treated with curcumin （0， 25， 50， 75 μmol·L-1） and 5-FU. Western blot was employed to determine the expression of proteins in the Janus kinase 1 （JAK1）/signal transducer and activator of transcription 1 （STAT1） /cyclin-dependent kinase inhibitor 1A （p21） pathway in HCT116 cells. The binding of STAT1 to p21 promoter region was detected by chromatin immunoprecipitation （ChIP）. Small interfering RNA （siRNA） was employed to measure the role of STAT1 in regulating the expression of p21 and that of JAK1 in regulating the activation of STAT1 by Western blot and cellular immunofluorescence， respectively. ResultCompared with the blank group， the HCT-116 cells treated with curcumin and 5-FU showed decreased viability （P<0.05）， increased proportions of cells in the G0/G1 phase （P<0.05）， decreased proportions of cells in the S phase and G2/M phase （P<0.05）， down-regulated protein level of phosphorylated p21 （P<0.05）， and up-regulated protein level of p21 （P<0.05）. Compared with the curcumin group， the p21 siRNA+ curcumin group presented decreased proportion of cells in G0/G1 phase （P<0.05）. Compared with the blank group， curcumin elevated the level of phosphorylated STAT1 （p-STAT1） （P<0.05）. Compared with the curcumin group， the curcumin + STAT1 siRNA group showcased up-regulated protein level of p21 in HCT116 cells （P<0.05）. The mechanism study showed that curcumin treatment enhanced the enrichment of STAT1 in the p21 promoter region （P<0.05） compared with the blank group. Compared with the blank group， curcumin up-regulated the level of phosphorylated JAK1 （p-JAK1） （P <0.05）. Compared with the curcumin group， the curcumin + STAT1 siRNA group demonstrated up-regulated protein levels of p-STAT1 and p21 in HCT116 cells （P<0.05）. ConclusionCurcumin may induce the cycle arrest of human colon cancer HCT116 cells by activating the JAK1/STAT1/p21 signaling pathway.

Ameliorative Effect and Mechanism of Qingwen Baiduyin on Lipopolysaccharide-induced Acute Lung Injury / 中国实验方剂学杂志

To investigate the therapeutic effect and mechanism of Qingwen Baiduyin on acute lung injury （ALI） in mice induced by lipopolysaccharide （LPS）. MethodA total of 144 C57BL/6 mice were randomly divided into the following groups： a normal group， a model group （LPS， 5 mg·kg-1）， a dexamethasone group （5 mg·kg-1）， and low-， medium-， and high-dose Qingwen Baiduyin groups （14.105， 28.21， 56.42 g·kg-1）. The mice were treated once daily for 5 days. One hour after the final administration， the ALI model was established by intratracheal instillation of LPS， and samples were collected at 6 h and 24 h after modeling. The arterial blood gas index of mice was analyzed. The total protein content， total cell count， Evans blue dye （EBD） content， and lung tissue wet-to-dry weight ratio （W/D） of bronchoalveolar lavage fluid （BALF） were measured. Hematoxylin-eosin （HE） staining was performed to assess the pathological changes in mouse lung tissue. Western blot was used to detect the expression levels of key proteins in the Janus kinase 1/signal transducer and activator of transcription 1/interferon regulatory factor 1 （JAK1/STAT1/IRF1） signaling pathway in lung tissue. ResultCompared with the normal group， the model group showed reduced arterial oxygen pressure （pO2）， oxygen saturation （SO2）， and lung tissue W/D （P<0.05， P<0.01）， increased carbon dioxide pressure （pCO2）， total protein content， total cell count， EBD content， interferon-γ （IFN-γ）， tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， chemokine CXC ligand 1 （CXCL1）， chemokine CXC ligand 2 （CXCL2）， chemokine CXC ligand 9 （CXCL9）， and chemokine CXC ligand 10 （CXCL10） content （P<0.05， P<0.01）， thickening of the alveolar walls， fusion of alveolar cavities， and infiltration of inflammatory cells in lung tissue， increased proportion of M1 macrophage polarization and lung cell apoptosis （P<0.05）， and increased protein expression levels of JAK1， phosphorylated JAK1 （p-JAK1）， inducible nitric oxide synthase （iNOS）， STAT1， phosphorylated STAT1 （p-STAT1）， IRF1， gasdermin D （GSDMD）， and mixed lineage kinase domain-like protein （MLKL） （P<0.05， P<0.01）. Compared with the model group， Qingwen Baiduyin significantly increased pO2， SO2， and lung tissue W/D （P<0.05， P<0.01）， improved the pathological changes in lung tissue， and reduced pCO2， total protein content， total cell count， EBD content， IFN-γ， TNF-α， IL-1β， CXCL1， CXCL2， CXCL9， and CXCL10 content， proportion of M1 macrophage polarization， and protein expression levels of JAK1， p-JAK1， iNOS， STAT1， p-STAT1， IRF1， GSDMD， and MLKL （P<0.05， P<0.01）. ConclusionQingwen Baiduyin can improve the lung inflammatory response and reduce lung cell apoptosis in mice with ALI by inhibiting the JAK1/STAT1/IRF1 signaling pathway， thereby exerting a lung-protective effect.

Molecular Mechanism of "Transmission Between Lung and Brain" of Influenza and Intervention Effect of Maxing Shigantang Based on JAK1/STAT1 Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo explore the molecular mechanism of "transmission between the lung and brain" of influenza based on Janus kinase 1/signal transducer and activator of transcription 1（JAK1/STAT1） signaling pathway and further investigate the intervention effect of Maxing Shigantang （MXSGT）. MethodA total of 100 SPF BALB/c mice were randomly divided into a normal group，a model group，an oseltamivir group （21.63 mg·kg-1·d-1），an antiviral granules group（3.9 g·kg-1·d-1）， and an MXSGT group（6.05 g·kg-1·d-1）， with 20 mice in each group. The pneumonia model was induced in mice except for those in the normal group by intranasal infection of influenza A virus（IAV）. Twenty-four hours after modeling，mice were treated with corresponding drugs， while those in the normal group and the model group received the same amount of normal saline by gavage， once a day for 3 and 7 days. The pathological changes in the lung and brain were observed by hematoxylin-eosin（HE）staining. The mRNA expression of IAV nucleoprotein（NP），JAK1， and STAT1 in the lung and brain was detected by real-time quantitative polymerase chain reaction（Real-time PCR）， and the protein expression of JAK1 and STAT1 in the lung and brain was detected by Western blot. Immunohistochemical method was used to detect the expression of phosphorylated（p）-STAT1 in the lung and brain tissues， and enzyme-linked immunosorbent assay（ELISA） was used to detect the serum levels of interleukin-1β（IL-1β） and interleukin-10（IL-10）. ResultCompared with the normal group， the model group showed obvious pathological changes in the lung tissues and cerebral cortex， increased relative mRNA expression of IAV NP in the lung （P<0.01）， elevated mRNA and protein expression of JAK1 and STAT1 in the lung and brain tissues （P<0.05，P<0.01），up-regulated expression level of p-STAT1 in lung tissues and cerebral cortex （P<0.05，P<0.01）， and increased serum level of IL-1β （P<0.05）. Compared with the model group， the MXSGT group showed alleviated pathological damage to lung tissues and cerebral cortex， decreased relative mRNA expression of IAV NP in lung tissues（P<0.01），reduced mRNA and protein expression levels of JAK1 and STAT1 in lung tissues and brain tissues（P<0.05，P<0.01）， and increased serum level of IL-10（P<0.01）. ConclusionThe abnormal activation of the JAK1-STAT1 signaling pathway may be one of the molecular mechanisms of "transmission between the lung and brain" of influenza. As an effective compound prescription against the influenza virus，MXSGT can alleviate the pathological damage of brain tissues in mice infected with IAV by regulating the level of cytokines mediated by this pathway.

Expression and clinical significance of signal transducer and activator of transcription-1, interferon regulatory factor-1,and interferon-γ in children with bronchial asthma / 中国小儿急救医学

Objective To study the expression profiles of signal transducer and activator of tran-scription-1(STAT1),interferon regulatory factor-1(IRF1),and interferon-γ(IFN-γ) in peripheral blood mononuclear cells in children with bronchial asthma,in order to further study their roles in the pathogenesis of asthma. Methods Thirty-eight children with acute exacerbation of bronchial asthma who visited pediatric outpatient clinic or hospitalized in pediatric department from September 2017 to May 2018 were enrolled as the asthma group. After standardized treatment,the 38 children with asthma were divided into the effective asthma treatment group(n＝23) and the ineffective asthma treatment group(n＝15),according to the treat-ment effect. Twenty-five healthy children were enrolled as the control group. RT-PCR and Western blot were used to measure the mRNA and protein expression levels of STAT1,IRF1,and IFN-γ in peripheral blood mononuclear cells. The correlations between the mRNA and protein expression of STAT1,IRF1,and IFN-γ were analyzed. Results The asthma group,the effective asthma treatment group and the ineffective asthma treatment group had significantly higher mRNA and protein expression levels of STAT1 and IRF1 than those of the control group (P<0. 05). The ineffective asthma treatment group had significantly higher mRNA and protein expression levels of IRF1 than those of the effective asthma treatment group (P<0. 05). The asthma group,the effective asthma treatment group and the ineffective asthma treatment group had significantly lower mRNA and protein expression levels of INF-γ than those of the control group (P<0. 05). Conclusion The expression level of STAT1 increases in children with asthma,while the expression level of IFN-γ decreases in those children. The expression level of IRF1 increases in children with asthma and is closely related to the treatment effectiveness.

Gain-of-function signal transducer and activator of transcription 1 deficiency / 中华实用儿科临床杂志

Chronic mucocutaneous candidiasis (CMC) is characterized by persistent or recurrent disease of the nails,skin,oral,or genital mucosae caused by candida albicans.CMC usually can occur in patients with T cell deficiencies,autosomal dominant hyper-immunoglobulin E(IgE) syndrome,interleukin(IL)-12p40 and IL-12 receptor β1 (IL-12Rβ1) deficiency,caspase recruiment domain 9 deficiency and autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy.CMC pathogenesis apparently involves the impairment of IL-17A,IL-17F and IL-22 immunity.Autosomal recessive IL-17RA deficiency and dominant-negative IL-17F deficiency are etiologies of pure isolated CMC (CMCD).Nearly half of patients with CMC had gain-of-function signal transducer and activator of transcription 1 mutations.These patients also had bacterial and virus infections,autoimmunity and inflammatory diseases,which show broad clinical heterogenity.

Mendelian susceptibility to mycobacterial disease / 中华实用儿科临床杂志

Mendelian susceptibility to mycobacterial diseases (MSMD)is a rare congenital disorder characterized by susceptibility to poorly virulent mycobacteria,such as Bacille Calmette-Guerin vaccine or non-tuberculous environmental mycobacteria.The interferon-γ'(IFN-γ)/interleukin-12 (IL-12) pathway is central to controlling mycobacterial infections,in which several genes had been identified.IFN-γ secretion is impaired in patients with IL-12p40 and IL-12 receptor β1 deficiency,where the response to IFN-γ is impaired in patients with IFN-γ receptor 1,IFN-γ receptor 2,and signal transducer and activator of transcription 1 deficiencies.Furthermore,germline mutations in the cytochrome b (-245) beta subunit,interferon regulatory factor 8,ubiquitin-like modifier,RORC and TYK2 have been identified as the genes which are responsible for MSMD.These patients do not generally have associated infections,apart from salmonellosis.Now,the pathogenesis,molecular,clinical,laboratory features,treatment and prognosis were described,in order to support the clues for pediatrician's clinical practice.

Effects of IL-33 on renal tubular injury in lupus nephritis through the JAK/STAT signaling pathway / 国际生物医学工程杂志

Objective To investigate the effects and mechanism of interleukin 33 (IL-33) on renal tubular injury in mice with lupus nephritis.Methods Twelve-week-old female MRL/lpr mice were randomly divided into model group,IL-33 group and solvent control group with 10 rats in each group.Ten female MRL/MP mice of the same age were used as normal control group.The mice in IL-33 group were intraperitoneally injected with 100 μL of phosphate buffer saline (PBS),containing 2 μg of recombinant mouse IL-33,once a day for 14 days.The mice in control group and the model group were intraperitoneally injected with the same dose of PBS.All the mice were sacrificed at 14 weeks of age.Serum creatinine (Cr) and urea nitrogen (BUN) concentrations were determined by serum separation.The urine in 24 hours was collected testing urinary protein creatinine ratio and urinary protein quantification.The contents of E-cadherin,α-SMA,and JAK/STAT pathway signaling proteins,including JAK2,p-JAK2,STAT1,and p-STAT1,were detected by Western blot.Results The BUN,urinary protein creatinine ratio and urine protein level of the IL-33 group were significantly higher than those of the model group (all P＜0.05).The expression of renal tubular epithelial cells o-SMA in the IL-33 group was higher than that in the model group,and the difference was statistically significant (P＜0.05).Compared with the model group,the expression of E-cadherin in the tubular epithelial cells of IL-33 group decreased and the expression of p-JAK2 and p-STAT1 protein increased,and the differences were all statistically significant (all P＜0.05).Compared with the model group,the levels of JAK2 and STAT1 in IL-33 group change little,and the differences were not statistically significant (all P＞0.05).Conclusions IL-33 can cause tubulointerstitial lesions in lupus mice,and its mechanism may be related to the activation of JAK/STAT pathway.

A case of chronic mucocutaneous candidiasis caused by STAT1 gain-of-function mutation and literature review / 中华实用儿科临床杂志

Objective To summarize the clinical characteristics,diagnosis and treatment of chronic mucocutaneous candidiasis(CMC).Methods The case diagnosed as CMC in the Department of Nephrology and Immunology of Shenzhen Children's Hospital in February 24,2014 was analyzed in terms of symptoms,signs,laboratory findings,gene tests and treatment process,and related literature was reviewed.Results The patient was a 14-year-old boy.The patient started to develop recurrent oral candida infection shortly after birth,then candida infection of skins and nails,which could be alleviated by antifungal agents,but easily relapsed.Since 4 years ago,autoimmune reactions such as autoimmune anemia,thrombocytopenia,leucopenia,proteinuria,and hypothyroidism had successively appeared,and cytopenia began to palliate after administering Glucocorticoid and Cyclosporin,but easily relapsed when the dosage was reduced.The genetic test showed the case was of signal transducer and activator of transcription 1 (STAT1) gain-of-function mutation.During the hospitalization,hemophagocytic syndrome (HPS) and stagnation of hematopoietic function successively occurred.The cytopenia did not improve and the patient suffered severe infection in spite of washing red blood cells 13 times and blood palate 3 times via infusion,together with high dosage of Dexamethasone and Cyclosporine.The peripheral blood cells and bone marrow gradually returned to normal after being treated by human granulocyte colony-stimulating factor combined with Dexamethasone and Cyclosporin.Retrieving the database in PubMed database,824 articles were found which were about CMC,and 39 of them were about the STAT1 gain-of-function mutation,including 120 cases.But there was only 1 domestic case in 2012,who was a three-year-old child,manifesting recurrent fungal infection of the skin.Conclusions STAT1 gain-of-function acquired mutation is one of the reasons that can lead to CMC.Autoimmune reactions prominently represented by cytopenia occur in a few patients with CMC.It should be alert on those who are with CMC and simultaneously with autoimmtne reaction of blood system.And gene tests facilitate the early diagnosis.

Comparison of improvement of two kinds of preventative administration of inter-leukin-27 on airway inflammation and impaired phosphorylation of STAT1 in asthma mice / 中国免疫学杂志

Objective:Animal models were set up to explore the best preventative intra-nasal administration of interleukin 27 (IL-27)to diminish allergic airway inflammation of asthma and the related molecular mechanisms. Methods:Ninety-six female C57/6J mice were randomly divided into four groups,a group of the control group,a group of asthma group,and two groups of the prevention group. Based on being sensitizing and challenging with Ovalbumin ( OVA ) in the asthma model, two kinds of IL-27 administration asthma animal models were set up,one of which was low-dose-multiple preventive administration before OVA sensitization,one was low-dose-multiple preventive administration after OVA sensitization but before OVA challenge. Sacrificed the mice after challenging and analyzed the IL-5 and IL-13 levels in supernatant of Broncho alveolar lavage fluid( BAL) using ELISA;and HE stain and inflammation score were done for the lungs. Sacrificed the mice before challenging and used the lungs to analyze the level of total signal transducer and activator of transcription-1(STAT1) protein and phos-STAT1 based on the method of Western blot. Results: In low-dose-multiple administration before sensitization preventions group,IL-27 inhibits the secretion of IL-5 and IL-13(P0. 05 ) . The phosphorylation of STAT1 was impaired in mice after OVA sensitization, and preventative administration of IL-27 before sensitization could reverse the impairment of STAT1, whereas another group had no obviously changes. Conclusion:Preventative administration of IL-27 before sensitization can attenuate the airway inflammation in the mouse asthma model via reversing the phosphorylation of STAT1,while the mice which has been sensitized resisting the inhibition of IL-27 due to the impairment of the phosphorylation of STAT1 and already committed Th2-CD4+T cells existed in sensitization-mice airway might be the reason for such IL-27 resistance.

Anti-inflammatory Activity of Fucoidan with Blocking NF-kappaB and STAT1 in Human Keratinocytes Cells

Fucoidan, a sulfated polysaccharide is found in several types of edible brown algae. It has shown numerous biological activities; however, the molecular mechanisms on the activity against atopic dermatitis have not been reported yet. We now examined the effects of fucoidan on chemokine production co-induced by TNF-alpha/IFN-gamma, and the possible mechanisms underlying these biological effects. Our data showed that fucoidan inhibited the TNF-alpha/IFN-gamma-induced production of thymus and activation-regulated chemokine (TARC) and macrophagederived chemokine (MDC) mRNA in human keratinocytes HaCaT cells. Also, fucoidan suppressed phosphorylation of nuclear factor kappa B (NF-kappaB) and activation of signal transducer and activator of transcription (STAT)1 in a dose-dependent manner. In addition, fucoidan significantly inhibited activation of extracellular-signal-regulated kinases (ERK) phosphorylation. These data indicate that fucoidan shows anti-inflammatory effects by suppressing the expression of TNF-alpha/IFN-gamma-induced chemokines by blocking NF-kappaB, STAT1, and ERK1/2 activation, suggestive of as used as a therapeutic application in inflammatory skin diseases, such as atopic dermatitis.

Influence of STAT1 on proliferation and IFN-βsensitivity of human non-small-cell lung cancer H1299 cells / 中国病理生理杂志

AIM:To investigate the effect of signal transducer and activator of transcription 1 ( STAT 1 ) on proliferation and interferon-β(IFN-β) sensitivity of human non-small-cell lung cancer H1299 cells.METHODS:STAT1 or EGFP gene was transfected into H1299 cells by the lentiviral vectors system.The cell number was counted under a mi-croscope and cell proliferation was tested by MTT assay.In addition, the cells transfected with STAT1 and EGFP were trea-ted with IFN-βand cell viability was measured by MTT assay.The protein levels of p-STAT1, ICAM-1 and PCNA were de-tected by Western blot.RESULTS: Over-expression of STAT1 inhibited H1299 cell proliferation (P<0.05).H1299 cells transfected with STAT1 gene had a higher sensitivity to IFN-βthan the control cells transfected with EGFP ( P <0.05).Overexpression of STAT1 increased the protein level of p-STAT1, and reduced IACM-1 expression in H1299 cells. Moreover, STAT1 enhanced STAT1 phosphorylation and downregulated the expression of PCNA in H1299 cells treated with IFN-β.CONCLUSION:STAT1 inhibits the proliferation and enhances the IFN-βsensitivity of non-small-cell lung cancer H1299 cells.

Effect of cyclosporine on the genetic expression of signal transducer and activator of transcription 1 in bladder cancer of rats and its clinical significance / 肿瘤研究与临床

Objective To investigate the effect of cyclosporine on signal transducer and activator of transcription 1 (STAT1) genetic expression on bladder cancer in rats induced by BBN and its clinical significance.Methods Twenty SD rats were divide into experimental group or control group randomly.Ten samples of SD rats bladder cancer induced with BBN and cyclosporine simultaneously as experimental group,and 10 samples of SD rats bladder cancer induced with BBN only as control.Real time RT-PCR and immunohistochemistry stain were used to detect STAT1 mRNA and protein level expressions of bladder cancer in rats respectively.Results The STAT1 mRNA median expression fold was 4.5 (2.1-6.6) in experimental group and 5.6 (3.4-8.5) in control group.The STAT1 protein expression were 5 cases with (-),3 cases (+),2 cases (++) in experimental group and 0 case (-),5 cascs (+),5 cases (++) in control group.The expression of STAT1 mRNA and protein level of bladder cancer between experimental group and control group were both significant different (P ＜ 0.05).Conclusions Cyclosporine may stimulate the growth and development of bladder cancer through changing expression of some genes like STATI,and STAT1 maybe become one of the targets of chemoprevention for post-transplantation bladder cancer.

Expression of janus kinase 1/signal transducer and activator of transcription 1 signaling pathway in cerulean stimulated pancreatic acinar cells / 中华胰腺病杂志

Objective To investigate the relationship between the activity of Janus kinase/signal transducer and activator of transcription (JAK/STAT) signaling pathway in cerulein-induced pancreatic acinar cell line AR42J. Methods The in vivo model of AP was induced by cerulean treated pancreatic acinar cell line AR42J, then RPM and AG490 were given for intervention. Western blot was used to determine theexpressions of JAK1 and phosphorylation JAK1 ( P JAK1 ) , STAT1, PSTAT1 and TNFα, IL-1β, IL-6. The expressions of IL-6, IL-1 β, and TNFα mRNA were measured by RT-PCR. Survival rate of cells was evaluated by trypan blue stain. Results The relative expressions of JAK1, P JAK1, STAT1, P STAT1 and TNF-o, IL-1β, IL 6 without cerulean treatment were 0.09 ±0.04,0.14 ±0.08,0.21 ±0.09,0.12 ±0.12,0.10 ±0.02,0.08 ± 0.03,0.02 ± 0.02. After cerulean treatment, the expressions of abovementioned protein increased in a time-dependant manner, the expressions at 24h were 0.53 ± 0.09,0.53 ± 0.13,0.56 ± 0.09,0.55 ± 0.10,0.25 ± 0.04,0.25 ±0.09,0.27 ±0.07, which were significantly higher than those in the control group (P ＜0.05). 2 4 h after RPM and AG 4 9 0 inhibition, the expressions of TNF-α, IL-1 β, IL-6 proteins significantly decreased to 0.17 ± 0.03and 0.17 ± 0.01,0.15 ± 0.05 and 0.14 ± 0.07,0.19 ± 0.04 and 0.19 ± 0.05; their expressions of mRNA significantly decreased ( P ＜ 0.05 ). The cell survival rates in RPM and AG490 treatment group were (72.4 ± 11.2) %, (69.7 ± 9.8 ) %, and in cerulein-stimulated cells (42.2 ± 12.3 ) % ( P ＜ 0.05 ).Conclusions The JAK1/STAT1 signaling pathway was involved in pancreatic inflammatory response with cerulein stimulation. Early treatment with inhibitors to the JAK1/STAT1 signaling pathway might control the inflammatory response in acute pancreatitis.

Effect of PIAS1 gene silencing on inflammatory response of pancreatic acinar cell / 中华胰腺病杂志

Objective To investigate the effect of protein inhibitor of activated signal transducer and activator of transcription 1 ( PIAS1 ) gene silencing on the inflammatory response of rat pancreatic acinar cell lines AR42J with cerulein stimulation, to study its role in the pathogenesis of acute pancreatitis.Methods The siRNA targeting PIASI was designed, synthesized, transfected into AR42J cells by lipofectmine 2000.24 h later, cerulean was added and cultured for another 24 h.Subsequent AR42J cells with cerulein stimulation were divided into 4 groups: cerulein, liposome, negative-siRNA and PIAS1-siRNA groups.In addition, a group with PBS was as control group.The activity of p38 mitogen- activated protein kinase (p38MAPK) was detected by western blotting.TNF-α, IL-1β, IL-6, matrix metalloproteinase (MMP) 9 expression were analyzed by RT-PCR and western blotting, respectively.Results The expression of p38MAPK in PIAS1-siRNA, negative-siRNA, liposome, cerulein,and control group was 1.93 ±0.11, 1.22 ±0.10, 1.30 ±0.17,1.32 ± 0.21, 0.12 ± 0.02;while the expression of phosphorylated p38MAPK was 2.10 ± 0.25, 1.36 ± 0.20,1.26 ±0.15, 1.23 ±0.25, 0.58 ±0.48, the expression in PIAS1-siRNA group was significantly increased when compared with other groups (P＜0.05).The levels of TNF-α, IL-1β, IL-6, MMP-9 mRNA were 1.66 ±0.15,1.66 ± 0.15,1.90 ±0.01, 1.56 ±0.20 in PIAS1-siRNA group, while the expression of protein was 2.06 ±0.37,2.20 ±0.34, 1.80 ±0.10, 1.17 ±0.05, which was markedly higher than those in other group (P ＜0.05).Conclusions PIAS1 gene silencing could enhance p38MAPK activity, and improve inflammatory mediator expression in pancreatic acinar cells with cerulein stimulation.

Effects of pulse methylprednisolone therapy on signal transducer and activator of transcription 1 activation of T cells in patients with severe systemic lupus erythematosus / 中华风湿病学杂志

Objective To investigate the effects of methylprednisolone pulse therapy on the expression of phosphorylated signal transducer and activator of transcription 1 (STATI) and DNA-binding activity of STATI in T cells in patients with severe systemic lupus erythematosus (SLE). Methods Six patients were included. Patients were given 0.5～1 g of methylprednisolone on 3 consecutive days. Western Blotting was conducted to explore the phosphorylated STATI expression and electrophoretic mobility shift assays (EMSA) were carried out to detect the DNA-biding activity of STATI. Results Methylprednisolone pulse therapy decreased phosphorylated STATI expression of T cells from patients with severe SLE. The expression of phosphorylated STATI decreased to about 30% 72 h after the methylprednisolone pulse therapy started (t=2.858, P<0.05). Methylprednisolone pulse therapy down-regulated DNA-biding activity of STATI of T cells in patients with severe SLE. The STATI DNA-biding activity was inhibited to about 40% 72 h after methy-Iprednisolone pulse, therapy started (t=3.058, P<0.05). Conclusion Phosphorylated STATI expression and DNA-binding activity of T cells is markedly decreased in patients after methylprednisolone pulse therapy, suggesting that inhibition of STATI signaling contributes to the clinical efficacy of this agent.

The relationship between cell apoptosis and Phospho-Ser~(727) Stat1 expression in the rat with experimental diffuse axonal injury / 中国法医学杂志

Objective To investigate the role of Stat1 in pathological process of nerve cells apoptosis induced by diffuse axonal injury (DAI) on rats. MethodsThe DAI model was established by using an injury model adapted from Marmarou et al. in 1994. All animals were divided into three groups, including control group, mock group and test group sacrificed on 6, 12, 24, 48, 72, 120 and 240 hours post injury (hpi). The paraffin-embedded sections of brain tissue were processed for HE staining and Bielschowsky’s silver method. Cell apoptosis was examined by flow cytometry and the expression of bax and bcl-2 were analyzed by RT-PCR. And Phospho-Ser727 Stat1 expression was examined by immunohistochemistry in different brain regions. ResultsThere was no brain contusion within HE staining, however, waving and enlargement of axons were observed within Bielschowsky’s silver method. The apoptotic rate of brain cells as well as PCR products ratio of bax to bcl-2 was highest at 24 hpi and decreased with time. An up-regulation of Phospho-Ser727 Stat1 at 6 hpi was discernible, and then reached the top at 24 hpi in cortex, cerebellum, brain stem and corpus callosum, and at 12 hpi in hippocampus. This increase was associated with the nerve cells apoptosis, r=0.921. In addition, the Phospho-Ser727 Stat1 positive cells were neurons and glial cells assessed from morphous. ConclusionsOur data indicate that Stat1 may contribute to the apoptosis of DAI on rats. In addition, the expression of Phospho-Ser727 Stat1 in glial cells suggested that glial cells may play an important role in the pathogenic mechanism of DAI.