Effects of benvitimod on levels of interleukin-4 and -10, interferon-γ and transforming growth factor-βin patients with atopic dermatitis / 中华皮肤科杂志

Objective To detect levels of interleukin?4(IL?4), IL?10, interferon?γ(INF?γ)and transforming growth factor?β(TGF?β)in the culture supernatant of peripheral blood mononuclear cells (PBMCs)from patients with atopic dermatitis(AD), and to evaluate regulatory effects of benvitimod on these cytokines. Methods PBMCs were isolated from 20 AD patients and 20 healthy controls. Then, PBMCs from AD patients were equally divided into 4 groups to be cultured with phosphate?buffered saline (PBS group), 400 nmol/L benvitimod solution (benvitimod group), 250 nmol/L dexamethasone solution (dexamethasone group) and 10 nmol/L tacrolimus solution (tacrolimus group), respectively. Enzyme?linked immunosorbent assay(ELISA)was performed to detect levels of IL?4, IL?10, INF?γand TGF?βin the culture supernatant of PBMCs. Results Compared with healthy controls, patients with AD showed significantly higher level of IL?4(83.4 ± 12.2 vs. 44.3 ± 5.7 pg/ml, P0.05). Compared with PBS, 400 nmol/L benvitimod could decrease the expression of IL?4(50.2 ± 10.1 vs. 83.3 ± 12.2 pg/ml, P 0.05)and INF?γ(9.56 ± 5.1 vs. 12.5 ± 2.3 pg/ml, P>0.05), but increase the expression of TGF?β(203.6 ± 15.3 vs. 178.9 ± 17.4 pg/ml, P>0.05)by PBMCs. In addition, no significant differences in the expression of IL?4, IL?10, INF?γ or TGF?β were observed between the benvitimod group and dexamethasone group or tacrolimus group(all P>0.05). Conclusion Benvitimod can regulate the expression of IL?4, IL?10, INF?γand TGF?βby PBMCs in patients with AD.

Polymorphism in the lymphotoxin-alpha gene, position +252 (rs909253), is not associated with preeclampsia development in Brazilian women / Polimorfismo no gene da linfotoxina alfa, posição +252 (rs909253), não está associado com o desenvolvimento de pré-eclâmpsia em mulheres brasileiras

PURPOSE: To investigate the frequencies of polymorphic allele and genotypes for the LT-α gene, position +252 (rs909253), in Brazilian women with preeclampsia.METHODS: This is a case-control study, in which 30 women with preeclampsia, classified according to the criteria of the National High Blood Pressure Education Program, and 115 women in the control group, with at least two healthy pregnancies, were selected. Peripheral blood was collected, and DNA was extracted, followed by genotyping, using specific primers and restriction analysis. The genotypes obtained were AA, AG and GG. Statistical analysis was performed using the χ2association test. The Hardy-Weinberg Equilibrium was tested using the Haploview Program.RESULTS: The results showed no association between genotypes and preeclampsia development (χ2=2.0; p=0.4). When the AG and GG genotypes were grouped according to allele G presence or absence (genotype AA), the data showed that the presence of allele G was not significantly different between cases (women with preeclampsia) and controls (χ2=0.0; p=1.0). The LT-α gene polymorphism, position +252 (rs909253), seems not to be an important candidate for the development of preeclampsia. Other inflammatory genes should be researched, and studies involving gene-environment interactions should be performed, in order to reach a better understanding of the etiology of the preeclampsia.

OBJETIVO: Investigar as frequências do alelo polimórfico e genótipos para o gene da LT-α, posição +252 (rs909253), em mulheres brasileiras com pré-eclâmpsia.MÉTODOS: Trata-se de um estudo caso-controle, em que 30 mulheres com pré-eclâmpsia, classificadas de acordo com os critérios do National High Blood Pressure Education Program, e 115 mulheres do grupo controle, com pelo menos duas gestações saudáveis, foram selecionadas. Amostra de sangue periférico foi colhida, e o DNA foi extraído, seguido pela genotipagem, usando iniciadores específicos e análise de restrição. Os genótipos obtidos foram AA, AG e GG. A análise estatística foi realizada utilizando-se o teste de associação χ2. O Equilíbrio de Hardy-Weinberg foi testado com o auxílio do programa Haploview.RESULTADOS: Os resultados não mostraram associação entre os genótipos e o desenvolvimento de pré-eclâmpsia (χ2=2,0; p=0,4). Quando os genótipos AG e GG foram agrupados de acordo com a presença do alelo G ou ausência (genótipo AA), os dados mostraram que a presença do alelo G não foi significativamente diferente entre casos (mulheres com pré-eclâmpsia) e controles (χ2=0,0; p=1,0). O polimorfismo no gene LT-α, posição +252 (rs909253), parece não ser um importante candidato para o desenvolvimento de pré-eclâmpsia. Outros genes inflamatórios devem ser pesquisados, e estudos envolvendo interações gene-ambiente devem ser realizados para que se possa alcançar um melhor entendimento da etiologia da pré-eclâmpsia.

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.

TNF-alpha and TNF-beta Polymorphisms are Associated with Susceptibility to Osteoarthritis in a Korean Population

BACKGROUND: The tumor necrosis factor (TNF) is believed to play an important role in the pathophysiology of osteoarthritis (OA). Evidence shows that genetic polymorphisms make substantial contributions to the etiology of OA. METHODS: We investigated the genotypes TNF-alpha and TNF-beta in 301 OA patients and 291 healthy subjects as controls. We employed a polymerase chain reaction-restriction fragment length polymorphism and a polymerase chain reaction-single strand conformation polymorphism assay to identify the genotypes TNFA -G308A and TNFB +G252A, respectively. RESULTS: For TNFA -G308A, the percentages of genotypes GG, AG, and AA were 26.3% (79/301), 62.5% (188/301), and 11.3% (34/301) in OA patients and 88.7% (258/291), 11.3% (33/291), and 0% (0/291) in controls. For TNFB +G252A, the percentages of genotypes GG, AG, and AA were 15.3% (46/301), 41.9% (126/301), and 42.9% (129/301) in OA patients and 12% (35/291), 52.6% (153/291), and 35.4% (103/291) in controls. There were significant differences in genotypes and alleles of TNFA -308 between OA patients and controls (p<0.0001) and in alleles of TNFB +252 (p=0.0325). The risk of OA was significantly higher for carriers of the TNFA -308A allele and the TNFB +252 AA homozygote (p=0.0224). CONCLUSIONS: The results suggest close relationships between TNFA -G308A and TNFB +G252A polymorphisms and individual susceptibility to OA in the Korean population.

The relationship among IL-10, TNF gene polymorphisms, Helicobacter pylori infection and gastroduodenal diseases in Hubei Han ethnic / 中华内科杂志

Objective To study the distribution of IL-10 and TNF gene polymorphisms in patients with gastroduodenal diseases in Hubei Han ethnic and their association with Helicobacter pylori (Hp) infection. Methods Six hundred and five patients with gastroduedenal diseases (196 chronic gastritis, 189 gastroduodenal ulcer and 220 gastric cancer) as well as 624 healthy controls were genotyped with PCR-RFLP method for IL-10-1082,-819,-592 and TNFα-308, lymphotoxin-α (LTα) Nco Ⅰ and AspH Ⅰ gene polymorphisms. Hp infection status was determined with ELLS& Results (1) There was significant difference of IL-10-1082 AG + GG genotype among the gastric cancer group with the non-malignant gastric diseases groups and healthy control group (P <0. 05). There was no significant difference of IL-10-592 and -819 gene polymorphisms among gastric cancer patients,non-malignant gastric disease patients and healthy controls (P>0. 05). The genotype frequencies of IL-10-819 were the same as those of IL-10-592. (2) Frequency of IL-10-1082 AG + GG genotype in gastric cancer patients with positive Hp was significantly higher than that in the other three groups (P < 0. 05). (3) Frequency of LTα Nco I AG genotype in gastric cancer patients with Hp infection was signiilcandy higher than that in Hp positive healthy controls (P < 0. 05). There were no other associations between TNFα-308, LTα Nco Ⅰ and AspH Ⅰ gene polymorphisms and Hp infection in gastroduodenal diseases. Conclusions (1) Allele AG + GG of IL-10-1082 was associated with gastric cancer in Han nationality of Hubei province. (2) IL-10-1082 AG + GG,LTct Nco ⅠAG heterozygous genotype may be associated with Hp infection in patients with gastric cancer in Han nationality of Hubei province.

Tumor Necrosis Factor and Lymphotoxin-alpha Gene Polymorphism in Korean Children with Type 1 Diabetes / 소아과

PURPOSE: Recently, it was reported that tumor necrosis factor (TNF) and lymphotoxin-alpha (LT-alpha) gene regions might be a susceptible loci to type 1 diabetes in Japanese. The purpose of this study was to investigate the association of TNF and LT-alpha gene polymorphisms with disease susceptibility in Korean children with type 1 diabetes. METHODS: Forty-nine Korean children with type 1 diabetes (29 girls and 20 boys) and 94 healthy Koreans were investigated in this study. Genotyping for -857T/C polymorphism in the TNF promoter region and LT-alpha gene polymorphism were performed by PCR-RFLP (restriction fragment length polymorphism). TNF promoter -1031C/T polymorphism was detected by allele-specific PCR. RESULTS: The distribution of the -857T/C and -1031C/T genotype in the TNF promoter region was not different between diabetic children and the controls. The frequency of TT genotype in the distribution of TNF -1031C/T polymorphism in diabetic children with diabetic ketoacidosis (DKA) at diagnosis was significantly lower than those without DKA (P< 0.05). No significant difference in the distribution of LT-alpha gene polymorphism was observed between diabetic children and the controls. There was no association between clinical characteristics of type 1 diabetes and LT-alpha gene polymorphisms. CONCLUSION: These results suggest that TNF promoter -857T/C and LT-alpha gene polymorphisms are not associated with susceptibility to type 1 diabetes in Korean children. TNF promoter -1031C/T polymorphism might be related to clinical manifestations (DKA) of type 1 diabetes.

Lymphotoxin-alpah gene polymorphism in patients with glomerulonephritis / 대한내과학회지

BACKGROUND: Lymphotoxin (LT)-alpha is a cytokine that has been involved in the inflammatory response, the pathogenesis of autoimmune disease, and the development and progression of renal injury in various glomerulonephritis. Glomerulonephritis is an important cause of end-stage renal disease requiring renal replacement therapy. A great body of evidence suggests that immunologic mechanisms and genetic factor such as gene polymorphism play an important role in the renal injury of glomerulonephritis. However, the role of LT-alpha gene polymorphism in primary glomerulonephritis is not clear yet. The purpose of this study is to examine whether there are the associations between LT-alpha gene polymorphisms and the development and progression of various primary glomerulonephritis. METHODS: A cross-sectional study of LT-alpha gene polymorphism by polymerase chain reaction with restriction fragment length polymorphism was performed on 190 patients with primary glomerulonephritis confirmed by renal biopsy and 249 controls. The gene polymorphism in the first intron of LT-alpha gene was detected by digestion with Nco1 restriction enzyme and subsequent electrophoresis. The clinical parameters at renal biopsy and the latest follow-up were collected. RESULTS: LTA1/LTA1 : LTA1/LTA2 : LTA2/LTA2 genotype distribution of the LT-alpha gene in the study population was 0.12 : 0.54 : 0.34, and the LTA1 : LTA2 allele frequency was 0.39 : 0.61. Thirty-one minimal change disease, 33 focal segmental glomerulosclerosis, 18 membranous glomerulonephritis, 16 membranoproliferative glomerulonephritis, and 92 InA nephropathy were included in this study. The distribution of LTA1/LTA1 : LTA1/LTA2 : LTA2/LTA2 was significantly different in the membranous glomerulonephritis (0.33 : 0.39 : 0.28, p<0.001) with control groups. But, there were no significant differences between the distributions of LT-alpha genotype in the patients with other glomerulonephritis groups. There were no significant differences in the age, systolic and diastolic blood pressure, serum creatinine, cholesterol, and proteinuria at renal biopsy among the three genotypes. There was no significant difference in the incidence of patients who started dialysis treatment or whose serum creatinine was double or more during the follow-up duration which was more than four years among the three genotypes. CONCLUSION: These data suggested that polymorphism in the LT-alpha gene may be associated with the development of primary membranous glomerulonephritis, however, these are not directly associated with the progression of renal injury in various glomerulonephritis.