Comparative Analysis of TNF-alpha, TNF-R1, and TNF-R2 in Patients with Low-impact Fractures Due to Osteoporosis / Análise comparativa de TNF-alfa, TNF-R1 e TNF-R2 em pacientes com fraturas de baixo impacto decorrentes de osteoporose

Abstract Objective To analyze the serum levels of TNF-alpha and its TNF-R1 and TNF-R2 receptors in the blood of patients with low-impact fractures due to osteoporosis, comparing between genders and with healthy patients. Methods The present study was conducted with a blood sample of 62 patients, divided into patients with osteoporosis and healthy patients. The results were obtained using the ELISA method. Cytokine concentrations were determined based on the absorbance values obtained. Results Serum TNF-alpha levels were undetectable in female patients, while in males they were found only in one patient, with no significant difference. Similar results were found in the analyses of TNF-R1 and TNF-R2 levels, a significant increase in levels of TNF-alpha receptors in the groups of patients with osteoporosis compared with the control groupinbothsexes.There wasnosignificant difference between the sexes in the dosage of both receptors within the group with osteoporosis. There was also a positive and significant correlation in the levels of TNF-R1 and TNF-R2 only in women. Conclusion The significant increase in TNF-R1 and TNF-R2 levels in women with osteoporosis suggest that the release and expression of these receptors may be contributing differently to the development of osteoporosis in men and women.

Resumo Objetivo Analisar os níveis séricos de TNF-alfa e de seus receptores TNF-R1 e TNF-R2 no sangue de pacientes com fraturas de baixo impacto, decorrentes de osteoporose, comparando entre os sexos e com pacientes saudáveis. Métodos Oestudofoi realizadocom amostradesanguede 62 pacientes,divididos em pacientes com osteoporose e pacientes saudáveis. Os resultados foram obtidos através do método de ELISA. As concentrações de citocinas foram determinadas com base nos valores de absorbância obtidos. Resultados Os níveis séricos de TNF-alfa foram indetectáveis nos pacientes do sexo feminino, enquanto no masculino encontrou-se somente em um paciente, não havendo diferença significativa. Encontrou-se resultados semelhantes nas análises dos níveis de TNF-R1 e TNF-R2, aumento significativo nos níveis dos receptores de TNF-alfa nos grupos de pacientes com osteoporose em comparação com o grupo controle, em ambos os sexos. Não houve diferença significativa entre os sexos na dosagem de ambos os receptores dentro do grupo com osteoporose. Houve ainda correlação positiva e significativa nos níveis de TNF-R1 e TNF-R2 apenas nas mulheres. Conclusão O aumento significativo nos níveis de TNF-R1 e TNF-R2 em mulheres com osteoporose sugerem que a liberação e expressão destes receptores pode estar contribuindo de maneira distinta no desenvolvimento da osteoporose em homens e mulheres.

Association of tumor necrosis factor receptor-associated protein 1 gene copy number variation with susceptibility and clinical characteristics of systemic lupus erythematosus / 中华风湿病学杂志

Objective To explore whether tumor necrosis factor receptor-associated protein 1 (TRAP1)gene copy number variation was associated with susceptibility and clinical characteristics of systemic lupus erythematosus (SLE).Methods The study enrolled 304 SLE patients and 391 healthy controls.They were used to investigate the association between TRAP1 gene copy number variation and SLE susceptibility.Then,304 SLE patients were divided into copy number=2 group and copy number＞2 group to study the association between TRAP1 gene copy number variation and disease activity or clinical characteristics of SLE.AccuCopyTM Kit was used to detect the TRAP1 gene copy number.Data analyses were performed by SPSS 10.01 software.The suitable method was selected among t test,rank sum test and x2 test for analysis based on the data type and distribution,univariate and multivariate logistic regression analysis were performed to investigate the associ-ation between TRAP1 gene copy number variation and susceptibility and clinical characteristics of SLE.Results The copy number variation of TRAP1 gene showed an association with the susceptibility to SLE crude OR=5.257,95％CI (1.108,24.937),P=0.037;the adjusted OR=5.578,95％CI (1.172,26.556),P=0.031].There was no association between TRAP1 gene copy number variation and SLE disease activity index (SLEDAI) score (Z=-0.117,P=0.907).The copy number variation of TRAP1 gene had a marginal association with skin lesions in SLE [OR=0.130,95％CI (0.016,1.069),P=0.058],but it disappeared after adjusting for potential confounders [OR=0.288,95％CI (0.029,2.831),P=0.286,PBH=0.808].There was no correlation between TRAP1 gene copy number variation and arthritis,alopecia,oral ulcers,fever,hematologic disorder,lupus nephritis as well as photosensitivity in SLE [x2=0.751,OR=1.234,95％CI (0.767,1.988),P=0.386].No multiplicative interaction was found between TRAP1 gene copy number variation and age or body mass index (BMI) [age:x2=0.751,OR=1.234,95％CI (0.767,1.988),P=0.386;BMI:x2=0.282,OR=1.172,95％CI(0.652,2.109),P=0.596].Conclusions The copy number variation of TRAP1 gene may be associated with susceptibility to SLE.Increased TRAP1 gene copy number may be a risk factor for SLE.

Expressions of TNF-α, TNFR1 and TNFR2 in different cervical lesions and their relationships with clinicopathological characteristics / 国际肿瘤学杂志

Objective To investigate the expressions of tumor necrosis factor-alpha (TNF-α),tumor necrosis factor receptor (TNFR)1 and TNFR2 in different cervical lesions and their relationships with clinicopathological characteristics.Methods Forty-one cases of cervical squamous cell carcinoma (CSCC) patients (CSCC group) treated in 254th Hospital of People's Liberation Army from January 2015 to December 2017 wereselected as the subjects.Forty-nine cases of high grde squmous intrepithelial lesion (HSIL) (HSIL group) and fifty cases of uterine myoma (normal group) were selected as control groups.The expression of TNF-α in cervical tissues of different lesions was detected by immunohistochemistry.The expressions of TNF-α mRNA,TNFR1 mRNA and TNFR2 mRNA were detected by quantificational real-time polymerase chain reaction (qRT-PCR).The expression level of TNFR1 and TNFR2 proteins are measured by Western blotting.The relationships between the expression level of TNF-α mRNA,TNFR1 mRNA and the clinicopathological characteristics of patients were analyzed.Results The positive rates of TNF-α in the normal group,the HSIL group and the CSCC group were 8.0％ (4/50),59.2％ (29/49) and 73.2％ (30/41).The difference between three groups was statistically significant (x2 =44.786,P ＜ 0.001).The positive rates of the HSIL group and the CSCC group were significantly higher than that in the normal group,the difference was statistically significant (x2 =29.175,P ＜ 0.001;x2 =40.883,P ＜ 0.001),but there was no statistical difference in the positive rates of TNF-α in CSCC group and HSIL group (x2 =1.934,P =0.164).The results of qRT-PCR showed that the expressions of TNF-α mRNA in normal group,HSIL group and CSCC group were 1.32 ± 0.21,3.64 ± 0.41 and 7.51 ± 1.42.The difference of TNF-α mRNA expression among three groups was statistically significant (F =655.800,P ＜ 0.001).The expressions level of TNF-α mRNA in HSIL group and CSCC group were significantly higher than that in normal group (t =31.747,P ＜ 0.001;t =51.012,P ＜ 0.001),and the expression level of CSCC group was significantly higher than that in HSIL group (t =20.039,P ＜ 0.001).The expression levels of TNFR1 mRNA in the normal group,the HSIL group and the CSCC group were 0.42 ± 0.13,0.89 ±0.21 and 2.23 ± 0.46.The relative expression of TNFR1 mRNA between the three groups was statistically significant (F =465.900,P ＜ 0.001).The expression levels of TNFR1 mRNA in group HSIL and CSCC were significantly higher than that in normal group (t =13.357,P ＜ 0.001;t =26.587,P ＜ 0.001),and the expression level of CSCC group was significantly higher than that in HSIL group (t =18.407,P ＜ 0.001).The expression of TNFR2 mRNA in the normal group,the HSIL group and the CSCC group were 0.38 ± 0.14,0.41 ± 0.11 and 0.44 ± 0.12.There was no significant difference between three groups (F =2.633,P =0.075).Western blottting showed that the expression intensity of TNFR1 in the normal group,the HSIL group and the CSCC group were 0.84 ±0.18,1.95 ±0.21 and 3.38 ±0.73,the difference was statistically significant (F =398.000,P ＜ 0.001).The expression intensity of TNFR1 in group HSIL and CSCC were significantly higher than that in normal group (t =18.273,P ＜ 0.001;t =39.894,P ＜ 0.001),and the expression in CSCC group was also significantly higher than that in group HSIL (t =22.357,P ＜ 0.001).The expression intensity of TNRF2 in normal group,HSIL group and CSCC group were 0.98 ± 0.15,1.02 ± 0.17,1.07 ± 0.21,and the difference was not statistically significant (F =2.938,P =0.056).The results of protein detection were in accordance with the results of mRNA detection.The expression of TNF-α mRNA in the CSCC tissues was related to the size of the tumor (t =-8.868,P ＜ 0.001),the degree of differentiation (t =-5.644,P ＜ 0.001),the clinical stage (t =-19.329,P ＜ 0.001),the depth of infiltration (t =-11.170,P ＜0.001),and lymph node metastasis (t =-8.339,P ＜ 0.001).The expression of TNFR1 mRNA was closely related to the tumor size (t =-13.309,P ＜ 0.001),degree of differentiation (t =-13.449,P ＜ 0.001),clinical stage (t =-12.949,P ＜0.001),depth of infiltration (t =-18.124,P ＜0.001),and lymph node metastasis (t =-20.506,P ＜ 0.001).Conclusion In cervical cancer tissues,the expression intensity of TNF-α and TNFR1 increased abnormally,while TNFR2 did not change significantly.The expressions of TNF-α and TNFR1 are positively correlated with the malignancy of cervical cancer.They are potential signals of cervical cancer and are expected to become new therapeutic targets.However,the activation of TNFR2 to downstream signaling pathway is significantly weaker than that of TNFR1.

Recombinant human tumor necrosis factor receptor type Ⅱ : IgG Fc fusion protein combined with methotrexate for the treatment of psoriasis and their effects on levels of interleukin-17A and tumor necrosis factor-α / 中华皮肤科杂志

Objective To evaluate the effect of recombinant human tumor necrosis factor receptor type Ⅱ:IgG Fc fusion protein (rhTNFR:Fc,trade name Etanercept) combined with methotrexate on levels of interleukin-17A (IL-17A) and tumor necrosis factor-α (TNF-α) in the serum and mononuclear cells of patients with moderate to severe plaque psoriasis.Methods A total of 30 patients with moderate to severe plaque psoriasis were enrolled from Department of Dermatology of Tenth People's Hospital of Tongji University between August 2014 and February 2016,and then were randomly and equally divided into Etanercept group and Etanercept + methotrexate group.The treatment lasted 24 weeks.Fifteen healthy blood donors served as healthy control group.Enzyme-linked immunosorbent assay (ELISA) and real-time quantitative PCR were performed to measure the serum levels and mRNA expression of IL-17A and TNF-α,respectively,in the patients of the above two groups before and after the treatment.Results Before the treatment,the serum levels of IL-17A and TNF-ct,as well as the mRNA expression of IL-17A and TNF-α in peripheral blood mononuclear cells (PBMCs),were all significantly higher in all the patients than in the healthy controls (all P ＜ 0.05).After the treatment,compared with the Etanercept group,the Etanercept + methotrexate group showed significantly lower serum levels of IL-17A (142.67 ± 14.82 vs.163.54 ± 23.18,P ＜ 0.05) and TNF-α (70.07 ± 25.02 vs.91.98 ± 14.62,P ＜ 0.05),as well as lower mRNA expression of IL-17A (1.12 ± 0.33 vs.1.56 ± 0.77,P ＜ 0.05) and TNF-α in PBMCs (2.50 ± 1.04 vs.3.61 ± 2.14,P ＜ 0.05).Conclusion Etanercept combined with methotrexate is superior to Etanercept alone in the treatment of psoriasis,and can reduce treatment duration and improve therapeutic effect,likely by down-regulating the expression of IL-17A and TNF-α.

Recombinant human tumor necrosis factor receptor type Ⅱ-IgG Fc fusion protein for treatment of occupational medicamentosa-like dermatitis induced by trichloroethylene / 中华劳动卫生职业病杂志

Objective@#To investigate the efficacy and safety of the recombinant human tumor necrosis factor receptor Ⅱ-IgG Fc fusion protein (rhTNFR: Fc, etanercept) for the treatment of occupational medicamentosa-like dermatitis induced by trichloroethylene (OMLDT) .@*Methods@#In September 2011 to February 2016, 12 patients with OMLDT were treated with etanercept 25 mg, subcutaneous injection, twice per week, doubling of first dose. The course of treatment was 6 weeks. The drug eruption area and severity index (DASI) score, the proportion of patients achieving a 50%, 75% and 90% reduction in DASI (DASI50, DASI75, DASI90) and the serum level of TNF-α were used to assess the efficacy at different times. Adverse reactions were also recorded and evaluated. The results were statistically analyzed by nonparametric Friedman test and repetitive measurement ANOVA using the software SPSS19.0.@*Results@#After 4 weeks treatment, the DASI score decreased form 56.33±7.02 to 0.50±0.91 (P<0.01) . The DASI50, DASI75 and DASI90 were all increased to 12 (100%) . The serum level of TNF-α decreased form (43.74±41.62) pg/ml to (3.03±0.47) pg/ml (P<0.01) . Statistically significant difference was observed from the above indexes. There were no adverse reactions in clinical application.@*Conclusion@#Recombinant human tumor necrosis factor receptor Ⅱ-IgG Fc fusion protein may be a safe and effective drug in the treatment of OMLDT.

Effects and underlying mechanisms of Cyr61 overexpression on cell apoptosis in early stage of ischemic acute kidney injury / 中华肾脏病杂志

Objective To detect the effect and mechanism of Cyr61 on the apoptosis of renal tissue caused by early stage of ischemic acute kidney injury (AKI). Methods 30 SD rats were randomized into 5 groups, including control group, AKI group, AKI+bicarbonate group, AKI+blank virus group, and AKI+over?expression Cyr61 virus group. After animal models were created for 2h, serum and renal tissue were collected from sacrificed animals. Expression level of TNF?α was determined by ELISA. HE staining was used to observe the histologic changes of renal tissues. The levels of NF?κB p65 and TNFR1 were measured by immunohistochemical method. RT?PCR and Western blotting assay were adopted to detect the mRNA and protein expression levels of NF?κB p65, TNFR1 and Caspase3. Results Compared with control group, AKI group, AKI+bicarbonate group, AKI+blank virus group, AKI+over?expression Cyr61 virus group had obvious kidney injury. The levels of TNF?α, the mRNA and protein expression levels of NF?κB p65, TNFR1 and caspase3 were markedly up?regulated. Over?expression of Cyr61 significantly attenuated the degree of pathological injury, numbers of apoptotic renal tubular epithelial cells and increased the degree of Scr. Although compared with other groups, the level of TNF?α in kidney tissue had no difference, there was obvious decreased protein level of NF?κB p65, while the increase of TNFR1 and Caspase3 protein was moderate. Conclusions During the early stage of AKI, over expression of Cyr61 could inhibit apoptosis, which may be related to the suppression of TNFR1 transcriptional expression and interference of TNF?αpathway. Its underlying mechanism therefore deserves further research.

The clinical significance of TNFR1 and TNFR2 in early diabetic nephropathy of type 2 diabetes / 中国医师杂志

Objective To explore the relationship of serum tumor necrosis factor receptor 1 (TNFR1),and TNFR2 with early diabetic nephropathy in type 2 diabetes patients.Methods A total of 70 patients with type 2 diabetes and 35 healthy person were selected from December 2013 to November 2014 in the Fifth Hospital of Datong,the urinary albumin excretion rate (UAER) was determined,and were divided into three groups:normal control group (n =35),simple diabetes group (n =35),and early diabetic nephropathy group (n =35).Enzyme-linked immuno sorbent assay (ELISA) method was used to determine TNFR1 and TNFR2.Results Compared to normal control group [TNFR1 (302.5 ± 60.0) pg/ml,TNFR2 (62.1 ±9.8)pg/ml],simple diabetes group [TNFR1 (515.1 ±78.1)pg/ml,TNFR2 (178.5 ±38.9)pg/ml],and early diabetic nephropathy group [TNFR1 (1 066.8 ± 205.6) pg/ml,TNFR2 (279.6 ± 37.1)pg/ml] were all increased;and early diabetic nephropathy group was higher than simple diabetes group (P＜ 0.05).The levels of TNFR1 and TNFR2 were positively correlated with age,course,glycated hemoglobin (HbA1c) levels(P ＜0.01),and were negatively correlated with high density lipoprotein cholesterol (HDL-C) and estimated glomerular filtration rate (eGFR) (P ＜0.01).The multiple stepwise regression analysis in eGFR had significant effects on TNFR 1 and TNFR 2 levels (P ＜ 0.01).Conclusions The levels of TNFR1,and TNFR2 in early diabetic nephropathy were increased.TNFR1 and TNFR2 may become new markers to evaluate early diabetic nephropathy.

Correlation of serum TNFR level with renal function injury in patients with T2DM and normal albumin ;excretion / 中国基层医药

Objective To discuss the correlation of serum TNFR level with renal function injury in patients with type 2 diabetes mellitus (T2DM)and normal albumin excretion.Methods 90 T2DMpatients with normal albu-min excretion(24h urine albumin excretion rate 90mL · min -1 · (1.72m2 )-1 ,eGFR 60 -90mL · min -1 · (1.72m2 )-1 and eGFR <60mL·min -1 ·(1.72m2 )-1 ,30 cases in each group.The serum TNFRs level and related clinical data were measured and analyzed statistically.Results The levels of serum TNFR1,TNFR2 were negatively correlated with eGFR(r =-0.428,-0.335,all P <0.05),and positively correlated with Scr(r =0.476,0.225,all P <0.05)in patients with T2DM and normal albumin excretion.The course of disease and the levels of BUN,Scr, CysC,α1 -MG,β2 -MG were increased as the degree of renal function injury.All these index had statistically signifi-cant differences in three groups(F =3.758,5.851,71.738,25.751,7.530,13.735,all P <0.05).Logistic regres-sion analysis showed that the levels of serum TNFR1,TNFR2 were independent risk factors for renal impairment in patients with T2DMand normal albumin excretion(OR =2.009,1.143,all P <0.05).Conclusion TNFRs may be involved in the development and progression of renal impairment,and may become a new conventional indicator for DN because of reflecting the degree of renal function injury in some extent in T2DMpatients with normal albumin excretion.

Effects of ginsenoside Rg1 on PARP-1 and TNFR1 expression in rat model of focal cerebral ischemia / 天津医药

Objective To explore effects of ginsenosides Rg1 on the expression of poly(ADP-ribose) polymerase-1 (PARP-1) and tumor necrosis factor receptor (TNFR) 1 in cortex cells after focal cerebral ischemia in rats. Methods Ninety healthy rats were randomly divided into sham-operative group, focal cerebral ischemia group, ginsenoside Rg 1groups (low, medium and high concentrations) and drug control group. Rats were intraperitoneally injected saline 45 mg/kg, saline 45 mg/kg+ginsenosides Rg1 10, 20 and 40 mg/kg, nimodipine 1 mg/kg 5 d before surgery, respectively. Focal cerebral isch?emia model was made by middle cerebral artery occluding in rats. The neurological deficit score and TTC staining were used to verify the success of the rat model. The expressions of PARP-1 and TNFR1 were evaluated by immunohistochemical meth?od and Western blot technique. Results There were obvious symptoms of neurological deficit and large pale infarct area in focal cerebral ischemia group compared with those of sham-operative group. There were higher percentages of neurological deficit score and infarct area in ginsenosides Rg1 groups and positive control group than those of sham-operative group, but which were lower than those of ischemia group (P<0.05). There were no significant differences between ginsenosides Rg1 groups and positive control group. The positive cells of PARP-1 and TNFR1 were higher in ginsenosides Rg1 low-dose group than those of sham-operative group and positive control group, while ones of medium and high-dose Rg1 group were higher than those of sham-operative group, and were lower than those of ischemia group (P<0.05). Compared with sham-op?erative group, PARP-1 and TNFR1 expression strips were significantly enhanced in ischemia group. Expression strips were higher in ginsenosides Rg1 low-dose group than those of sham-operative group. Expression strips were higher in ginsen?osides Rg1 medium-dose group than those of sham-operative group, but which were lower than those of ischemia group, and ones of high-dose group were lower than ischemia group (P<0.05). Conclusion Ginsenoside Rg1 shows protective effects on focal ischemia injury, which may be related with down-regulation of the expression of PARP-1 and TNFR1.

Significance of circadian gene Period2 expression in epithelial ovarian cancer and the effect of gene overexpression on growth of ovarian cancer xenografts in nude mice / 中华妇产科杂志

Objective To study the significance of circadian gene Period2 expression in epithelial ovarian cancer tissues and the effect of gene overexpression on the growth of ovarian cancer xenografts in nude mice. Methods Twenty-two cases of ovarian cancer paraffin specimens in the First Hospital of Shanxi Medical University (ovarian cancer group) were chosed during Jau. 2010 to Dec. 2013, including 8 cases of stageⅠ, 8 cases of stageⅡ, and 6 cases of stageⅢ, while 6 cases of benign ovarian epithelial tumor paraffin specimens were selected as control (benign tumor group). Period2 gene were detected by real-time quantitative PCR and western blot methods in different stages of ovarian cancer tumor tissues. Established theovarian cancer xenografts in nude mice with ovarian cancer cell line SKOV3, and they weredivided into 3 groups (n=8), including the recombinant plasmid group, empty plasmid group and control group. Using gene transfection technique to transfer Period2 gene into tumor tissues, tested the expression of Period2 mRNA in tumor tissues by real-time quantitative PCR after transfection into all nude mice, monitoredthetransplant tumor growth and calculating the tumor inhibition rate,detected the antiapoptotic gene BRE, apoptosis related tumor necrosis factor receptor (TNFR1) andtumor suppressor gene NIX in tumor tissues by real-time PCR and western blot in different groups. Results (1)The expression level of Period2 mRNA in tumor tissues amongovarian cancer group stageⅠ,ⅡandⅢwere respectively 2.59±0.50, 0.47± 0.08 and 0.42 ± 0.08, but benign tumor group was 6.59 ± 1.05. The expression level of Period2 protein in ovarian cancer group stage Ⅰ,Ⅱ and Ⅲ were respectively 0.835 ± 0.087, 0.412 ± 0.035 and 0.199 ± 0.031, while benign tumor group was 0.874 ± 0.094. The expression level of Period2 mRNA and protein in benign tumor group was higher than those in ovarian cancer group stageⅠ,ⅡorⅢ(P<0.01). With ovarian cancer stage increased, the expression of Period2 mRNA and protein were decreased or absent (P<0.05).(2)Two weeks after transfection, the expression level of Period2 mRNA in recombinant plasmid group tumor tissue was significantly higher than those in the empty plasmid group or the control group (6.11±0.56 vs 0.50±0.09 vs 0.44 ± 0.08, respectively;P<0.01), the transplanted tumor volume of recombinant plasmid group was significantly less than those in empty plasmid group or the control group[ (486±70) mm3 vs (835±106) mm3 vs (846 ± 110) mm3, respectively;P<0.01], the tumor inhibition rate of the recombinantin plasmid group was as high as 42.9%, that was significantly higher than those in the empty plasmid group and the control group (3.8% and 0, respectively;P<0.05).(3)The expression level of BRE mRNA and protein in transplanted tumor tissues in the recombinant plasmid group were significantly lower than those in empty plasmid group and the control group;the expression level of TNFR1 and NIX were significantly higher than those in the empty plasmid group and the control group (all P<0.05). Conclusions Period2 mRNA and protein expression are absent in ovarian cancer of advanced stage. Transfection and stable expression of Period2 gene could slow down the growth of ovarian cancer, and the tumor inhibition rate could be significantly increased. Period2 gene may promote ovarian cancer cells apoptosis through inhibition of BRE gene expression and promoting TNFR1, NIX gene expressionto exert anti-tumor effect.

Screening and identification of a novel small-molecule TNFβinhibitor / 天津医药

Objective To explore a novel and highly specific small-molecule TNFβinhibitor by using computer-aid?ed virtual screening and cell-based assays in vitro. Methods Computer-aided drug design and virtual screening were used to design and identify chemical compounds that targeted TNFβbased on the crystal structure of the TNFβ-TNFR1 com?plex. The effect of the small-molecule compound against TNFβ-induced cytotoxicity of L929 cells was detected by MTT as?say, and the efficacy of the compound to inhibit TNFβ-induced apoptosis of L929 cells was determined by flow cytometry as?say. The impact of the compound on L929 cell cycle was examined by Propidium Iodide (PI) staining and flow cytometry, and the influence of the compound on TNFβ-triggered signal pathway was analyzed by Western blot assay and Ultra VIEW VOX 3D Live Cell Imaging System. Results No.35 compound (named as C35 thereafter) could effectively inhibit TNFβ-induced cell death in a dose dependent manner, and the half-maximum inhibition concentration (IC50) was 8.19μmol/L. Furthermore, C35 had lower cytotoxicity and minimal effect on L929 proliferation. Here we further revealed that C35 could affect TNFβ-induced apoptotic pathway by blocking the activation of Caspase 3, and markedly reduce L929 cell apoptosis induced by TNFβ. Conclusion A novel TNFβsmall-molecule inhibitor was identified by combining computer-aided virtual screening with functional assays, and which could block TNFβ-triggered apoptotic pathway and efficiently inhibit the cell death in?duced by TNFβ.

IL-10 and ET-1 as biomarkers of rheumatic valve disease / IL-10 e ET-1 como biomarcadores de doença valvar reumática

Objective: To evaluate the immunological profile and gene expression of endothelin-1 (ET-1) in mitral valves of patients with rheumatic fever originated from a reference service in cardiovascular surgery. Methods: This was a quantitative, observational and cross-sectional study. Thirty-five subjects (divided into four groups) participated in the study, 25 patients with chronic rheumatic heart disease and ten control subjects. The mean age of the sample studied was 34.5 years. Seventeen of them (48.58%) were male and 18 (51.42%) were female. Inflammatory cytokines (TNF-α, IL-4 and IL-10) were measured and ten mitral valves of patients who underwent first valve replacement were collected for determination of gene expression of endothelin-1 by real time PCR. Results: Among the groups studied (patients vs. controls), there was a statistically significant difference in IL-10 levels (P=0.002), and no differences in other cytokines. Expression of endothelin-1 was observed in 70% of samples. Quantitatively, average of ET-1 expression was 62.85±25.63%. Conclusion: Inflammatory cytokine IL-10 participates in the maintenance of chronicity of rheumatic fever in patients who underwent valve replacement and those who are undergoing medical treatment. The expression of endothelin-1 in heart valve lesions in patients undergoing mitral valve replacement confirms its association with inflammatory activity in rheumatic fever. .

Objetivo: Avaliar o perfil imunológico e a expressão gênica de endotelina-1 em valvas mitrais de pacientes com febre reumática, originados de um serviço de referência em cirurgia cardiovascular. Métodos: Este foi um estudo quantitativo, observacional e transversal. Trinta e cinco indivíduos (divididos em quatro grupos) participaram do estudo, 25 deles com doença cardíaca reumática crônica, além de 10 controles. A média de idade da amostra estudada foi de 34,5 anos. Dezessete (48,58%) dos indivíduos eram homens, e 18 (51,42%) eram mulheres. Foram medidas algumas citocinas inflamatórias (TNF-α, IL-4 e IL-10) e coletadas 10 valvas mitrais de pacientes que se submeteram a primeira troca valvar para determinação da expressão gênica de endotelina-1 pelo PCR real-time. Resultados: Entre os grupos estudados (pacientes e controles), observou-se diferença estatisticamente significante em relação aos níveis de IL-10 (P=0,002), sem diferenças nas outras citocinas. Em relação à endotelina-1, foi observada sua expressão em 70% das amostras. Quantitativamente, a expressão média de endotelina-1 foi de 62,85±25,63%. Conclusão: A citocina inflamatória IL-10 participa da manutenção da cronicidade da febre reumática em pacientes que se submeteram a troca valvar e naqueles que estão em tratamento médico. A expressão de endotelina-1 nas lesões em valvas cardíacas de pacientes que foram submetidos à troca valvar mitral confirma sua relação com a atividade inflamatória na febre reumática. .

The Research Progress of Tumor Necrosis Factor Alpha in Multiple Sclerosis and Remyelination / 天津医药

Tumor necrosis factor (TNF)-αis a pleiotropic inflammatory cytokine, which is produced chiefly by acti?vated macrophages. Two forms of TNF-α, soluble and transmembrane, can bind tumor necrosis factor receptor (TNFR) 1 or TNFR2, respectively. Recently, a concept has emerged that TNF-α/TNFR pathway plays an important role in the pathogene?sis of multiple sclerosis and remyelination. TNFR1 induces death of oligodendrocytes via death receptor-mediated apoptosis, which leads to demyelination or other neurodegenerative changes. However, TNFR2 has a positive effect on multiple sclero?sis. It facilitates the proliferation and differentiation of oligodendrocyte precursor cells, thus promoting remyelination.

Association of tumor necrosis factor-α receptor gene single nucleotide polymorphism in patients with ankylosing spondylitis / 中华风湿病学杂志

Objective To investigate the value of tumor necrosis factor (TNF)-α receptor gene,TNFRSF1A+36A/G(rs767455) and-383A/C(rs2234649),TNFRSF1B+196T/G(rs1061622) single nucleotide polymorphism (SNP) for the susceptibility to ankylosing spondylitis (AS) and the relationship between SNP and AS.T test,Chi-square test,and ANOVA were used for statististical analysis.Methods Two hundred and fifteen patients who had definite diagnosis of AS and 216 healthy blood donors were involved in this study.SNPs of TNF-α receptor gene:TNFRSF1A +36A/G(rs767455),-383A/C(rs2234649) and TNFRSF1B+196T/G (rs1061622) were detected with the ligase detection reaction (LDR-PCR) method.Results ① Distribution frequencies of A alleles(86.8％,91.5％) and G alleles (13.2％,8.5％) of TNFRSF1A(rs767455) in AS and controls were significantly different with each other (x2=4.627,P=0.0315),while the distribution frequency in group of homozygotes (AA or GG genotype) in AS and controls were 74.6％(150/201) and 83.9％(177/211),the frequencies in group of heterozygotes (AG) were 25.4％ (51/201) and 16.1％(34/211)(x2=5.390,P=0.020).Frequency of alleles and the genotypes of TNFRSF1A (rs2234649) and TNFRSF1B (rs1061622) between AS and control group were similar(P＞0.05).It also demonstrated that TNF-αreceptor gene haplotype (rs1061622T-rs2234649A-rs767455G) carriers apparently increased the susceptibility to AS (11.5％ vs 6.9％)(OR:1.753,95％CI:1.078～2.852,P=0.022).② Analysis of variance found that the duration of morning stiffness (F=3.168,P=0.044) and peripheral joint tenderness counts (F=4.598,P=0.011) among the three genotype groups of TNFRSF1B (rs1061622) in patient with AS were evidently differed with each other.Bath AS functional index (BASFI) among different genotype groups of TNFRSF1A (rs2234649) in AS had remarkable diversity (F=5.783,P=0.004).None of above indicators among groups of different genotypes of TNFRSF1A (rs767455) in AS were uniform (P＞0.05).③ Forty-four patients were treated with TNF-α antagonist (entanercept),25 mg,subcutaneous injection,twice weekly for 3 months,then followed with Sulfaslazine (SASP) 2.0 g/d and Celecoxib 0.4 g/d for another 9 months.ASAS20 was the primary endpoint for the evaluation of therapeutic effect at the visit of 3 month and 12 month.No associations were found between SNP and short or long term outcome of treatment with TNF-α antagonist in AS (P＞0.05).Conclusion TNFRSF1A (rs767455) SNP correlates with susceptibility to AS in Anhui Han local patients.Carriers of TNF-α receptor gene haplotype (rs1061622T-rs2234649A-rs767455G) may increase the susceptibility to AS.SNP of TNFRSF1B (rs1061622) is associated with disease activity in AS,while SNP of TNFRSF1A(rs2234649)relates to functional index of the disease.There is no association between SNP of TNFRSF1A / TNFRSF1B and short or long term outcome of treatment with TNF-α antagonist in AS.

Decoy receptor3 in rheumatoid arthritis patient and its clinical relevance / 中华风湿病学杂志

Objective To test serum decoy receptor 3 (DcR3) concentration in patients with rheumatoid arthritis (RA),and analyze its clinical significance.Methods The serum from 180 RA patients,29 osteoarthritis (OA) patients,60 systemic lupus erythematosus (SLE) patients,30 Sj(o)gren's syndrome (SS) patients and 100 healthy donators were selected.The serum DcR3 concentrations of the two groups were detected by enzyme-linked immunosorbent assay (ELISA).The correlation between serum DcR3 levels and clinical features of patients with RA were investigated.T test,x2 test and Spearman's correlation analysis were used for statistical analysis with software SPSS 16.0.Results Serum DcR3 levels in RA patients [(202±261)ng/ml] were significantly higher than those of the healthy controls [(32±31) ng/ml,t=5.33,P＜0.01],OA patients [(90±33)ng/ml,t=1.998,P＜0.01],SLE patients [(41±56)ng/ml,t=3.19,P＜0.01],and SS patients [(25±31) ng/ml,t=2.50,P＜0.01].The DcR3 cut value was 45.86 ng/ml for the diagnoses of RA,the sensitivity and specificity was 0.991 and 0.765 respectively.The levels of serum DcR3 in RA patients were related with rheumatoid factor (RF) IgM,RF-IgG,anti-cyclic citrullinated peptide antibodies (anti-CCP) and C3.Conclusion Serum DcR3 level is significantly elevated in RA and it is significantly associated with immunological index of RA including RF-IgM,RF-IgG,C3 and anti-CCP antibody,which may be a factor for unfavorable prognosis of RA patients.

The clinical investigation of OX40/OX40L from lymphocytes of patients with Graves' disease after 131I therapy / 中华核医学与分子影像杂志

Objective To investigate the clinical value and the expression of OX40/OX40L from lymphocytes of patients with Graves' disease (GD) before and after 131I therapy.Methods The expression of OX40/OX40L on T,B lymphocytes and the percentage of lymphocyte subsets were analyzed using immunofluorescence staining and flow cytometry in 32 patients with GD before and after 131I therapy.Twenty-five healthy subjects were considered as the control group.The data between GD patients and controls were compared with two-sample t test.The correlation between the expression of OX40/OX40L and the function of thyroid (FT3,FT4 and TRAb level) was performed with linear correlation analysis.Results Compared with the healthy subjects,the percentage of CD4+ T cells and ratio of CD4+/CD8+ were decreased ((34.36 ±6.73)％ vs (45.60-±5.72)％,t=2.634; 1.24±0.26 vs 1.84±0.37,t=2.125,both P＜0.05).Whereas the percentage of CD19+ B cells in patients with GD were significantly higher than that in healthy subjects ((21.42 ±3.92)％ vs (15.35 ± 3.15)％,t =2.653,P ＜0.05).The expression of OX40 increased in CD4+ T lymphocytes ((12.92 ± 2.26) ％) and OX40L expression was up-regulated in CD19+ B lymphocytes ((15.33 ± 3.75)％) of patients with GD,compared with the healthy subjects ((4.19 ±1.45) ％ and (8.64 ± 1.73) ％,respectively,t =2.112 and 2.467,both P ＜ 0.05).The expression of OX40 in CD4+ T lymphocytes ((7.65 ± 1.64) ％) and OX40L in CD19+ B lymphocytes ((11.50 ±2.72)％) were increased after 131I therapy(t =2.795,2.393,both P ＜0.05).The thyroid functions of 32 GD patients were improved.The levels of FT3,FT4 and TRAb decreased significantly after 131I treatment (from 20.79 ±6.05 to 4.53 ± 2.54,from 49.65 ± 10.12 to 13.69 ± 4.35,and from 24.78 ± 8.46 to 11.74 ±6.19,respectively; t =2.219,2.441 and 2.293,all P＜0.05).The levels of FT3 and FT4 were positively correlated with the percentage of OX40 expression in CD4+ T lymphocytes (r =0.65 and 0.73,both P ＜ 0.05).TRAb was positively correlated with CD19+ OX40L + B lymphocytes (r =0.76,P ＜ 0.05).Conclusions OX40/OX40L may play an important role in the activation of lymphocytes,the production of antibodies,and participate in the pathogenesis and progression of GD.131I therapy not only damages most of the thyroid follicular epithelial cells by its β ray,but also facilitates GD improvement by regulating T and B lymphocyte functions.

Herpes Virus Entry Mediator Signaling in the Brain Is Imperative in Acute Inflammation-Induced Anorexia and Body Weight Loss

BACKGROUND: Reduced appetite and body weight loss are typical symptoms of inflammatory diseases. A number of inflammatory stimuli are responsible for the imbalance in energy homeostasis, leading to metabolic disorders. The herpes virus entry mediator (HVEM) protein plays an important role in the development of various inflammatory diseases, such as intestinal inflammation and diet-induced obesity. However, the role of HVEM in the brain is largely unknown. This study aims to investigate whether HVEM signaling in the brain is involved in inflammation-induced anorexia and body weight loss. METHODS: Food intake and body weight were measured at 24 hours after intraperitoneal injection of lipopolysaccharide (LPS) or intracerebroventricular injection of recombinant mouse LIGHT (also called tumor necrosis factor receptor superfamily 14, TNFSF14), an HVEM ligand, into 8- to 10-week-old male C57BL/6 mice and mice lacking HVEM expression (HVEM-/-). We also assessed LPS-induced change in hypothalamic expression of HVEM using immunohistochemistry. RESULTS: Administration of LPS significantly reduced food intake and body weight, and moreover, increased expression of HVEM in the hypothalamic arcuate nucleus. However, LPS induced only minor decreases in food intake and body weight in HVEM-/- mice. Administration of LIGHT into the brain was very effective at decreasing food intake and body weight in wild-type mice, but was less effective in HVEM-/- mice. CONCLUSION: Activation of brain HVEM signaling is responsible for inflammation-induced anorexia and body weight loss.

Predictive Value of Conjointly Examined IL-1ra, TNF-R I, TNF-R II, and RANTES in Patients with Primary Glomerulonephritis

Interleukin-1 receptor antagonist (IL-1ra), tumor necrosis factor soluble receptors (sTNF-R) type I and II, and regulated upon activation, normal T-cell expressed and secreted (RANTES) play an important role in the modulation of primary glomerulonephritis (GN) course. The aim of the study was to assess whether pre-treatment measurements of IL-1ra, sTNF-R, and RANTES assessed conjointly may be useful as predicting factors in patients with GN. In 84 patients (45 males and 39 female) serum concentration (pg/mL) and urinary excretion (pg/mgCr) of cytokines were measured. After 12 months of therapy with steroids and cyclophosphamide the patients were divided into two subgroups: Responders (R) and Non-Responders (NR) according to the treatment results. The urinary IL-1ra, TNF-RI and RII were significantly higher in R than NR (1,732 vs 646 with P < 0.001, 13.1 vs 6.3 with P = 0.005, and 33.6 vs 14.4 with P = 0.012). The urinary RANTES excretion was increased in NR (79.6 vs 28.5; P < 0.001). The multivariable analysis showed that if conjointly assessed, only urinary IL-1ra, TNF-R I and R II, RANTES with 85% probability pointed the feature remission (R). In conclusion, the urinary excretion of IL-1ra, TNF-R I and R II, and RANTES examined conjointly are effective in predicting favorable response to immunosuppressive treatment in patients with GN.

Effect of tumor necrosis factor receptor-1 on neurovascular regeneration after cerebral ischemia in mice / 国际脑血管病杂志

Objective To investigate the effect of tumor necrosis factor receptor 1 (TNFR1) in angiogenesis and neurogenesis during cerebral ischemia in mice.Methods Twenty-four wild-type and 24 TNFR1 knockout mice were randomly divided into either a sham operation group or a focal cerebral ischemia group (n =12 in each group).5-bromodeoxyuridine (BrdU) was injected intraperitoneally at day 3 after cerebral ischemia and sham operation.At day 7 and 28 after cerebral ischemia,the double-label immunofluorescence staining of glucose transporter-1(Glut-1)/BrdU was used to evaluate the angiogenesis surrounding the areas of infarction.A labeled BrdU was used to detect the neural stem cell proliferation in the subventricular zone.Double-labeled doublecortin (DCX)/BrdU and neuronal nuclei antigen (NeuN)/BrdU were used to detect the migration and survival of neural stem cells,respectively.Results Under the normal condition,there was no significant difference in angiogenesis and the number of BrdU-positive cells in the subependymal zone (SVZ) between the wild-type (418.000 ± 28.404) and TNFR1 knockout (528.000 ± 60.597) mice (t =-1.644,P =0.131).At day 7 after cerebral ischemia,the number of Glut-1/BrdU-positive cells in the TNFR1 knockout mice was significantly less than that in the wild-type mice (14.833 ± 2.182 vs.27.5 ± 4.209) (t =2.672,P =0.023),and the number of DCX/B3rdU-positive cells was also significantly less than that in the wild-type mice (163.000 ± 11.106 vs.257.168 ± 12.213) (t =5.705,P =0.000).At day 28 after cerebral ischemia,the number of NeuN/BrdU-positive cells in the TNFR1 knockout mice was significantly less than that in the wildtype mice (6.000 ± 0.577 vs.11.000± 1.571) (t=2.988,P=0.014).Conclusions TNFR1 may play a promoting role in the neurovascular reggeneration in late cerebral ischemia.

The study of the effect on IgM-RF, IgG-RF, IgA-RF of patients with rheumatoid arthritis hy the treatment of recombinant human tumor necrosis factor receptor Fc fusion protein / 中华风湿病学杂志

Objective To observe the effect of recombinant human tumor necrosis factor receptor Fc fusion Protein (rhTNFR:Fc) treatment on IgM-RF,IgG-RF,IgA-RF of patients with rheumatoid arthritis.Methods A randomized,active-comparator controlled,parallel group study were conducted.110 patients were enrolled and were randomly divided to the treatment group,in which patients were treated with twice weekly subcutaneous injection of rhTNFR:Fc (25 mg) (rhTNFR:Fc treatment group,n=55),and the MTX froup,in which MTX (the mean dosage was 15 mg/week) (MTX group,n=55) for 24 weeks.Blood routine,IgM-RF,IgG-RF,IgA-RF,and disease activity score 28 (DAS28) were monitored.Student's t-test was used for statistical analysis.Results The level of IgM-RF decreased significantly in the rhTNFR:Fc treatment group 24 week(29±16) U/ml later.However,the level of IgG-RF(145±20) U/ml,IgA-RF(153±34) U/ml increased significantly in the rhTNFR:Fc group,and the level of IgG-RF (62±14) U/ml,IgA-RF (66-±19) U/ml decreased significantly in the MTX group.Conclusion Although rhTNFR:Fc,is effective in treating the clinical symptoms of RA,it seems to affect RF producing-B cells either directly or indirectly.