Objective: To investigate the effect of microRNA (miR-148b) targeting decoy receptor 3 (DcR3) on macrophage polarization in sepsis. Methods: Experimental study. From December 2019 to December 2022, serum microRNA expression was detected in 3 patients with sepsis and 3 healthy controls in the clinical laboratory of Songjiang Hospital Affiliated to Shanghai Jiao Tong University School of Medicine. Phorbol 12-myristate 13-acetate (PMA) was used to induce the differentiation of human acute monocytic leukemia cells THP-1 into macrophages, and then lipopolysaccharide (LPS) was added to stimulate the establishment of a sepsis cell model, and the expression changes of miR-148b and DcR3 were detected by RT-PCR and Western blot. Overexpression of DcR3 was used to detect the expression levels of TNF-α, CD163 and IL-10 in macrophages stimulated by LPS (100 ng/ml). Overexpression of miR-148b was used to observe the changes of molecular markers of macrophage polarization. The targeting regulation effect of miR-148b on DcR3 was determined by dual-luciferase reporter assay. t test was used to analyze whether there were statistical differences among the groups. Results: The expression of miR-148b was down-regulated (P<0.05) and the expression of DcR3 was up-regulated (P<0.01) in THP-1 macrophages stimulated by LPS. Overexpression of DcR3 inhibited the expression of TNF-α (P<0.05) and promoted the expression of CD163 (P<0.01) and IL-10 (P<0.01). When miR-148b mimics was added, the opposite effect was observed. The dual-luciferase reporter assay confirmed that miR-148b targets and binds to DcR3, inhibiting its transcription and expression. The results of flow cytometry showed that DcR3 could reverse the promoting effect of miR-148b on the CD86/CD163 ratio of macrophages (P<0.05). Conclusion: miR-148b inhibits the expression of DcR3, thereby inhibiting M2 polarization in LPS-stimulated macrophage cells.
Humans ; Interleukin-10 ; Lipopolysaccharides/pharmacology* ; Macrophages ; MicroRNAs/genetics* ; Receptors, Tumor Necrosis Factor, Member 6b/metabolism* ; Tumor Necrosis Factor-alpha

Objective: To investigate the effect of microRNA (miR-148b) targeting decoy receptor 3 (DcR3) on macrophage polarization in sepsis. Methods: Experimental study. From December 2019 to December 2022, serum microRNA expression was detected in 3 patients with sepsis and 3 healthy controls in the clinical laboratory of Songjiang Hospital Affiliated to Shanghai Jiao Tong University School of Medicine. Phorbol 12-myristate 13-acetate (PMA) was used to induce the differentiation of human acute monocytic leukemia cells THP-1 into macrophages, and then lipopolysaccharide (LPS) was added to stimulate the establishment of a sepsis cell model, and the expression changes of miR-148b and DcR3 were detected by RT-PCR and Western blot. Overexpression of DcR3 was used to detect the expression levels of TNF-α, CD163 and IL-10 in macrophages stimulated by LPS (100 ng/ml). Overexpression of miR-148b was used to observe the changes of molecular markers of macrophage polarization. The targeting regulation effect of miR-148b on DcR3 was determined by dual-luciferase reporter assay. t test was used to analyze whether there were statistical differences among the groups. Results: The expression of miR-148b was down-regulated (P<0.05) and the expression of DcR3 was up-regulated (P<0.01) in THP-1 macrophages stimulated by LPS. Overexpression of DcR3 inhibited the expression of TNF-α (P<0.05) and promoted the expression of CD163 (P<0.01) and IL-10 (P<0.01). When miR-148b mimics was added, the opposite effect was observed. The dual-luciferase reporter assay confirmed that miR-148b targets and binds to DcR3, inhibiting its transcription and expression. The results of flow cytometry showed that DcR3 could reverse the promoting effect of miR-148b on the CD86/CD163 ratio of macrophages (P<0.05). Conclusion: miR-148b inhibits the expression of DcR3, thereby inhibiting M2 polarization in LPS-stimulated macrophage cells.
Humans ; Interleukin-10 ; Lipopolysaccharides/pharmacology* ; Macrophages ; MicroRNAs/genetics* ; Receptors, Tumor Necrosis Factor, Member 6b/metabolism* ; Tumor Necrosis Factor-alpha

Humans ; Haploinsufficiency ; NF-kappa B/genetics* ; Mutation ; Tumor Necrosis Factor alpha-Induced Protein 3/genetics*

OBJECTIVETo explore the relationship between the polymorphism of TNF-&alpha; gene 308, 238 locus and the susceptibility to pneumoconiosis.

METHODSEighteen published case-control studies about TNF-&alpha; gene 308, 238 locus polymorphism and pneumoconiosis susceptibility were searched out from sino-foreign databases from January 1994 to December 2010. Meta-analysis was applied on the published research to calculate the pooled OR value (95%CI) and stratified analyze the types and species of pneumoconiosis.

RESULTSEleven of the published research articles were selected into the analysis, including 10 research focusing on TNF-&alpha; gene 308 locus, with 1408 cases and 1639 controls in total. The meta-analysis showed that comparing with Gln/Gln carriers, Arg/Arg, Arg/Gln, Gln/Arg + Arg/Arg carriers were 1.89-fold (95%CI: 1.10 - 3.24), 1.53-fold (95%CI: 1.25 - 1.87), and 1.56-fold (95%CI: 1.28 - 1.90) more susceptible to pneumoconiosis, respectively. The stratified analysis showed that among coal workers, the TNF-&alpha; gene 308 locus Arg/Arg, Arg/Gln, Gln/Arg + Arg/Arg carriers were separately 2.29-fold (95%CI: 1.22 - 4.29), 1.56-fold (95%CI: 1.20 - 2.03), 1.64-fold (95%CI: 1.28 - 2.11) more susceptible to pneumoconiosis than Gln/Gln carriers; and among Asian people, the TNF-&alpha; gene 308 locus Gln/Arg, Gln/Arg + Arg/Arg carriers were separately 1.58-fold (95%CI: 1.28 - 1.95) and 1.57-fold (95%CI: 1.28 - 1.94) more susceptible to pneumoconiosis than the Gln/Gln carriers. Four case-control research focus on the study of TNF-&alpha; gene 238 locus, including 391 cases and 391 controls in total. The analysis showed that comparing with the non-carriers, TNF-&alpha; gene 238 locus Arg/Arg, Arg/Gln, Gln/Arg + Arg/Arg carriers were 6.03-fold (95%CI: 1.35 - 26.97), 1.87-fold (95%CI: 1.07 - 3.30) and 2.36-fold (95%CI: 1.37 - 4.07) more susceptible to pneumoconiosis.

CONCLUSIONTNF-&alpha; gene 308, 238 locus Arg/Arg, Gln/Arg, Gln/Arg + Arg/Arg carriers are more susceptible to pneumoconiosis.

Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Pneumoconiosis ; genetics ; Polymorphism, Genetic ; Tumor Necrosis Factor-alpha ; genetics

Genetic Predisposition to Disease ; Humans ; Pneumoconiosis ; genetics ; Polymorphism, Genetic ; Tumor Necrosis Factor-alpha ; genetics

Oral lichen planus (OLP) is a common chronic inflammatory disease of the oral mucosa. The prevalence rate of OLP in adults is 0.5%-2%. The etiology and pathogenesis of OLP are still unclear. The pathogenesis of OLP may be related to the genetic polymorphism of some genes. Currently, the gene families, including tumor necrosis factor, interferon, interleukin, enzyme, and receptor, have been extensively studied. This work reviews related studies on gene polymorphism of OLP.
Adult ; Humans ; Lichen Planus, Oral/genetics* ; Mouth Mucosa ; Polymorphism, Genetic ; Tumor Necrosis Factor-alpha/genetics*

OBJECTIVETo investigate the relationship between the presence of the TNF2 allele and plasma concentrations of tumor necrosis factor-alpha (TNF alpha) and soluble TNF receptor (sTNF-R) with the development of acute severe pancreatitis (ASP) and severe sepsis.

METHODSGenomic DNA was prepared from peripheral blood leukocytes. The TNF1 and TNF2 biallelic polymorphisms were identified by analyzing NcoI-digested DNA fragments obtained from PCR products. Plasma levels of TNF alpha and sTNF-R were measured by EASIA.

RESULTSThe overall TNF2 allele frequency in ASP patients was comparable to that found in healthy volunteers (29.2% vs. 29.3%, P > 0.05). Severe sepsis occurred in 26 of 72 patients. Patients with severe sepsis showed a significantly higher prevalence of TNF2 than those without (46.2% vs. 19.6%, P < 0.05). Plasma TNF alpha, sTNF-R I, and sTNF-R II levels were (36 +/- 31) pg/ml, (5.4 +/- 3.5) ng/ml, and (11.2 +/- 7.8) ng/ml, respectively, in patients with severe sepsis, and (31 +/- 25) pg/ml, (4.6 +/- 3.8) ng/ml, and (8.8 +/- 6.6) ng/ml in non-severe sepsis subjects. Differences in TNF levels were not statistically significant between patients with ASP and control group (P > 0.05). Moreover, there was no correlation between TNF2 allele frequency and TNFalpha levels [(37 +/- 31) pg/ml vs. (31 +/- 25) pg/ml in TNF2 group and TNF1 group, respectively, P > 0.05].

CONCLUSIONSOur results suggest that there is no relationship between ASP and the TNF2 allele, but that the TNF2 allele is associated with a susceptibility to severe sepsis as a result of ASP.

Acute Disease ; Adult ; Female ; Humans ; Male ; Middle Aged ; Pancreatitis ; genetics ; Polymorphism, Genetic ; Receptors, Tumor Necrosis Factor ; blood ; Sepsis ; genetics ; Tumor Necrosis Factor-alpha ; analysis ; genetics

OBJECTIVE: To determine the polymorphism in +252 site of tumor necrosis factor-beta(TNF-beta) gene in patients with or without endometriosis, to evaluate the levels of TNF-alpha and TNF-beta in the serum with or without endometriosis, to explore the relation between polymorphism of TNF-beta gene and the genetic susceptibility of endometriosis, and to explore the pathogenic mechanism of endometriosis at gene level. METHODS: By polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method, polymorphism on +252 site of TNF-beta gene was measured in 82 patients with endometriosis (the endometriosis group) and 80 patients without endometriosis (the control group). With the sandwich-enzyme-linked immunosorbent assay (ELISA), the levels of TNF-alpha and TNF-beta in the serum of the two groups were determined. RESULTS: The TNF-beta level in the serum in the endometriosis group with TNF-beta gene +252 site AA genotype significantly increased, compared with GG genotype (t=2.029, P<0.05); while TNF-alpha and TNF-beta level in the serum had no statistical significance in patients with other genotypes in TNF-beta gene +252 site in the endometriosis group and the control group. CONCLUSION TNF-beta gene +252 site AA genotype might be enhance TNF-beta level in the serum of patients with endometriosis.
Adolescent ; Adult ; Endometriosis ; blood ; genetics ; Female ; Humans ; Lymphotoxin-alpha ; blood ; genetics ; Polymorphism, Genetic ; Tumor Necrosis Factor-alpha ; blood ; Young Adult

OBJECTIVES: To investigate whether necrostatin-1 (Nec-1) can protect islet cells from the damage induced by TNF-α. METHODS: After isolation and purification, the neonatal porcine islet cell clusters (NICCs) were divided into 3 groups (islets 10 000 IEQ/group): a Nec-1 group (Nec-1+TNF-α was added to the culture medium), a TNF-α group (TNF-α was added to the culture medium), and a control group (pure medium). The number of cells was observed after 48 h of co-culture. The cell death was evaluated by AO/EB staining. Insulin secretion and DNA of islets were detected by chemiluminescence and nucleic acid quantitative analysis. RT-PCR assay was used to examine the mRNA expressions of insulin gene, glueogan gene and somatostatin gene. Flow cytometry analysis was used to detect the viability of B cells. RESULTS: The number of islets in Nec-1 group, TNF-α group and the control group were (8 425±2 187), (4 325±778), and (7 122±1 558) IEQ, respectively. Compared to the other two groups, the number of dead cells in TNF-α group was greatly increased. The insulin/DNA values in the Nec-1 group, TNF-α group and blank control group were (13.21±3.15), (2.47±0.45), and (7.44±0.97) mIU/mg, respectively. Compared to the TNF-α group and the control group, the mRNA relative expression levels of insulin gene (6.73±1.07), glucagon gene (10.13±1.98), somatostatin gene (8.57±1.11) were significantly increased in the Nec-1 group (all <0.05), the rate of live cells (97.32±1.87)% and live B cells (90.86±3.68)% were increased significantly in the Nec-1 group (all <0.05). CONCLUSIONS TNF-α can induce neonatal porcine islet cells damage, which is attenuated in the presence of Nec-1. Nec-1 can increase the content of endocrine cells in NICCs.
Animals ; Imidazoles ; Indoles ; Insulin ; Islets of Langerhans ; Swine ; Tumor Necrosis Factor-alpha ; genetics

OBJECTIVE: To observe the effect of moxibustion on skin lesions and immune inflammatory response in psoriasis mice, and to explore the possible mechanism of moxibustion for psoriasis. METHODS: A total of 32 male BALB/c mice were randomly divided into a normal group, a model group, a moxibustion group and a medication group, 8 mice in each group. Psoriasis model was induced by applying 5% imiquimod cream on the back for 7 days in the model group, the moxibustion group and the medication group. At the same time of model establishment, the moxibustion group was treated with suspension moxibustion on skin lesions on the back, 20 min each time, once a day; the medication group was treated with 1 mg/kg methotrexate tablet solution by gavage, once a day. Both groups were intervened for 7 days. The daily changes of skin lesions were observed, and the psoriasis area and severity index (PASI) score was evaluated; the histopathological changes of skin lesions were observed by HE staining; the positive expression of proliferating cell nuclear antigen (PCNA) and T lymphocyte surface marker CD₃ were detected by immunohistochemistry; the expression level of serum interleukin (IL) -17A was detected by ELISA, and the relative expressions of tumor necrosis factor-α (TNF-α), IL-1β and IL-6 mRNA in skin lesions were detected by real-time PCR. RESULTS: The increased and hypertrophy scale, dry skin, red and swollen epidermis and obvious infiltration were observed in the model group, and each score and total score of PASI were higher than those in the normal group (P<0.01). The scale score, infiltration score, and total score of PASI in the moxibustion group were lower than those in the model group (P<0.01); the infiltration score and total score of PASI in the medication group were lower than those in the model group (P<0.01, P<0.05). The inflammatory cell infiltration in the model group was obvious, and the thickness of epidermal layer was increased compared with that in the normal group (P<0.01); the inflammatory cell infiltration and Munro micro abscess were decreased in the moxibustion group and the medication group, and the thickness of epidermal layer was decreased compared with that in the model group (P<0.01). Compared with the normal group, the positive cell number of PCNA and T was increased (P<0.01), and the body mass was decreased, and the spleen index was increased (P<0.01), and the expression of serum IL-17A and the relative expression of TNF-α, IL-1β and IL-6 mRNA in the skin lesions was increased in the model group (P<0.01). Compared with the model group, the positive cell number of PCNA and T was reduced (P<0.01), and the spleen index and the relative expression of TNF-α, IL-1β and IL-6 mRNA were reduced (P<0.01) in the moxibustion group and the medication group; the body mass of mice in the moxibustion group was higher than that in the model group (P<0.01); the content of serum IL-17A in the medication group was lower than that in the model group (P<0.01); the relative expression of TNF-α, IL-1β mRNA in the moxibustion group was higher than that in the medication group (P<0.01). CONCLUSION Moxibustion could effectively improve the scale and infiltration of skin lesions in psoriasis mice. Its mechanism may be related to inhibiting inflammatory response and regulating immunity.
Animals ; Imiquimod ; Male ; Mice ; Moxibustion ; Psoriasis/therapy* ; Skin ; Spleen ; Tumor Necrosis Factor-alpha/genetics*