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Objective To investigate the therapeutic effects and mechanisms of interleukin-10 (IL-10) gene-modified dendritic cells (DC-IL-10) in mice with liver fibrosis. Methods DC-IL-10 was constructed in vitro, the phenotype and function of which were evaluated by flow cytometry. BALB/c mice were treated with intraperitoneal injection of carbon tetrachloride(CCl4)to establish liver fibrotic model. DC-IL-10 was administrated via tail vein. Animals were divided into 4 groups including normal dendritic cell (DC) control , liver fibrosis only, negative lentiviral transfection DC (DC-mock) and DC-IL-10. Liver function, cytokine secretion, T lymphocyte differentiation and liver histomorphology were tested. Real-time PCR and western blot were used to analyze the effect of DC-IL-10 on Wnt/β-catenin signaling pathway and its role in liver fibrosis. Results When compared with DC control and DC-mock, the expression of DC-IL-10 surface stimulating molecules (major histocompatibity complex-Ⅱ, CD80, CD86) were significantly decreased (F=14.708, 22.503, 12.595, respectively, all P<0.05), and DC-IL-10 significantly inhibited T lymphocyte proliferation (F=50.295, P<0.05). When compared with liver fibrosis group, serum alanine aminotransferase and aspartate transaminase were decreased in DC-IL-10 treated group (all P<0.05), other parameters including inflammatory factors (tumor necrosis factor α, IL-6, IL-1β) reduced (all P<0.05), the proportion of regulatory T cells (Treg) increased (F=6.742, P<0.05), pathological damage improved, the expression of Wnt3a, α-SMA and β-catenin mRNA and protein significantly reduced in DC-IL-10 treatment group(all P<0.001). Conclusions DC-IL-10 induces elevation of Treg for immune tolerance, as well as inhibition of inflammatory response, block of Wnt/β-catenin signaling pathway, which translates into improvement of liver fibrosis.
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OBJECTIVE Gestational rhinitis(GR) is a relatively common condition in women during pregnancy.Patients with GR often complain of nasal obstruction and rhinorrhea.The exact mechanism of GR are not clear.Safe and effective treatments for this disease have not been found to date.The aim of this study is to find an appropriate treatment method for GR.METHODS Thirty patients with GR were randomly divided into two groups.There were 15 patients in hypertonic saline group with a mean age of 28.73 years (range 24-31 years),and 15 patients in normal saline group with a mean age of 25.93 years (range 24-31 years).Hypertonic saline group was treated using 3.0％ saline with a temperature of 40℃ nasal irrigation,and normal saline group was treated with 0.9％ saline with a temperature of 40℃ nasal irrigation.The duration of the intervention period was 4 weeks.Visual Analog Scale(VAS) was used to evaluate the nasal symptoms including nasal obstruction and rhinorrhea,and the health-related quality of life was assessed with the 12-item Short Form Health Survey version 2.0(SF-12v2).Contents of histamine(HIS) and acetylcholinesterase(ACHE) in nasal lavage fluid(NLF) was assessed before and after 4-week treatment in the two groups in the study.RESULTS There were 28 patients completed the study.The total VAS scores of nasal symptoms including nasal obstruction and rhinorrhea decreased,and SF-12v2 score increased in the hypertonic saline group after 2-week,3-week and 4-week interventions.Furthermore,ACHE in NLF was also increased after 4-week treatment,but HIS showed no statistical changes.The VAS scores of nasal obstruction and rhinorrhea and SF-12v2 score after 2-week,3-week and 4-week interventions,and the contents of HIS and ACHE in NLF after 4-week treatment showed no statistical differences in the normal saline nasal irrigation group.There were statistical differences in the VAS scores of nasal obstruction and rhinorrhea,SF-12v2 score and ACHE in NLF after 4-week treatment,and no significant differences in the content of HIS in NLF between the 2 groups.CONCLUSION Hypertonic saline nasal irrigation is a safe and effective treatment for GR.
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Objective@#To observe the therapeutic effect of simple 3.0% saline nasal irrigation and combined treatment of 3.0% saline nasal irrigation and budesonide nasal spray for vasomotor rhinitis (VMR), and explore the long-term effect for VMR. Through examination of levels of substance P (SP) and mucin (MUC)5B in nasal lavage fluid, the mechanisms of nasal irrigation treatment for VMR was discussed.@*Methods@#One hundred and one patients from Department of Otorhinolaryngology Head and Neck Surgery, Huashan Hospital of Fudan University with VMR were randomly divided into 4 groups. The number of patients was 24 in control group, 25 in budesonide nasal spray treatment group (budesonide group), 25 in nasal irrigation treatment group (nasal irrigation group) and 27 in budesonide nasal spray + nasal irrigation group (combined treatment group). Control patients were left untreated. Budesonide group was under budesonide nasal spray treatment, nasal irrigation group was treated using 3.0% saline with a temperature of 40℃ and combined treatment group was given both treatments. The duration of the intervention period was 3 months (90 days). Visual Analog Scale (VAS) was used to evaluate nasal symptoms, and the health-related quality of life was assessed using the 12-item Short Form Health Survey version 2.0 (SF-12v2). Enzyme-linked immunosorbent assay (ELISA) was used to assess the contents of SP and MUC5B in nasal lavage fluid before and after 3-month treatments in budesonide and nasal irrigation group in the study. MUC5B in nasal lavage fluid after the SP challenge and anticholinergic drug intervention in control group were also evaluated with ELISA.@*Results@#Nighty out of 101 patients completed the study. In the budesonide and combined treatment group after relevant interventions, the total VAS score of nasal symptoms decreased (5.91±0.21 vs 3.82±0.15, 6.18±0.17 vs 3.92±0.15, t value was 8.193, 10.060, respectively, all P<0.05) and SF-12v2 score increased (146.00±1.23 vs 152.30±0.97, 146.00±1.08 vs 155.40±0.90, t value was 3.982, 6.697, respectively, all P<0.05), with both scores showed no significant differences in the nasal irrigation group (5.96±0.17 vs 5.72±0.15, 146.10±1.17 vs 147.00±0.94, t value was 1.038, 0.607, respectively, all P>0.05) after the first month. In the budesonide and combined treatment group after relevant interventions, the total VAS score of nasal symptoms decreased (5.91±0.21 vs 5.05±0.15, 6.18±0.17 vs 5.10±0.12, t value was 3.374, 5.351, respectively, all P<0.05) and SF-12v2 score increased (146.00±1.23 vs 150.90±0.76, 146.00±1.08 vs 153.60±0.94, t value was 3.373, 5.343, respectively, all P<0.05), with both scores showed no significant differences in the nasal irrigation group (5.96±0.17 vs 5.78±0.17, 146.10±1.17 vs 148.10±0.80, t value was 0.716, 1.438, respectively, all P>0.05) after the second month. By the end of the third month, in nasal irrigation and combined treatment group, the VAS score was diminished (5.96±0.17 vs 4.80±0.12, 6.18±0.17 vs 4.44±0.13, t value was 5.485, 8.264, respectively, all P<0.05) and SF-12v2 score was elevated (146.10±1.17 vs 150.80±0.96, 146.00±1.08 vs 152.90±0.85, t value was 3.163, 5.008, respectively, all P<0.05), but there were no significant differences in budesonide group (5.91±0.21 vs 5.68±0.18, 146.00±1.23 vs 148.40±0.85, t value was 0.819, 1.587, respectively, all P>0.05). Additionally, SP in nasal lavage fluid decreased and MUC5B showed no statistical changes in budesonide group after three months, however, SP showed no any changes and MUC5B reduced significantly in nasal lavage fluid in nasal irrigation group. Furthermore, the anticholinergic drug could not decrease the concentration of MUC5B after the SP challenge in nasal cavity in control group.@*Conclusions@#The therapeutic effect of simple nasal irrigation with 3.0% saline or combined treatment of 3.0% saline nasal irrigation and nasal corticosteroids is superior to simple nasal corticosteroids. Nasal corticosteroids plays a role in the inhibition of sensory nerve endings in nasal mucosa, but neurotransmitter plays a limited role in the pathogenesis of VMR.
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Objective To investigate the impact of suppressor of cytokine signaling 1 (SOCS1) overexpression on dendritic cells (DC) functions and its therapeutic effect on acute liver failure (ALF) in mice.Methods Bone marrow derived dendritic cells (BMDC) from C57BL/6 mice were transfected with lentivirus encoding SOCS1 and negative control lentivirus at a MOI=50, and labeled as DC-SOCS1and DC-VNG, respectively after 96 hours of successful transduction.Then DCs were stimulated with lipopolysaccharides(LPS)1 mg/L and collected for flow cytometry analysis of surface costimulatory molecules, allogeneic mixed lymphocyte reaction (MLR) and western blot test of Janus kinase (JAK)/signaling transducers and activators of transcription (STAT) pathway.Afterwards, 90 mice were randomly assigned into 4 groups including 12 in normal control group, 26 in ALF group, 26 in treatment groups with DC-SOCS1 and 26 with the treatment of DC-VNG.All were received tail vein injection with normal saline, modified DC-VNG and DC-SOCS1 suspended in normal saline, respectively.Twelve hours after injection, LPS (10 μg/kg)/D-GaIN (600 mg/kg) were injected intraperitoneally to induce ALF model.The mortality, serum levels of alanine aminotransferase (ALT) and aspartate transaminase (AST), liver pathology and proportion of splenic regulatory T cells of each group were observed.Means in different groups were compared with one-way ANOVA analysis.Categorical variables were analyzed with x2 test.Variables were examined with normality test and homogeneity of variance with LSD test.Results The results of mixed lymphocyte reaction (MLR) revealed that T cell proliferation ratio in DC-SOCS1 group with mixture ratio of 100∶1 were (25.87±0.38)%, which was lower than that of mixture ratio of 10∶1 in the mDC group ([84.29±3.25]%) with statistical significance (x2=49.821, P<0.01);interleukin (IL)-10 concentration was higher than that in mDC group with mixture ratio of 10∶1 with statistical significance (F=20.112, P<0.05);IL-6 concentration was also lower with statistical significance (F=47.718, P<0.05).Compared to imDC, expression of JAK2 (t=0.525,0.523 and 0.489, respectively, all P<0.01), signal transduction factors and activation of transcription factors-1 (STAT1) (t=0.442,0.400 and 0.402, respectively, all P<0.01) and SOCS1 (t=0.322,0.363 and 1.090, respectively, all P<0.01) of mDC, DC-VNG and DC-SOCS1 after LPS stimulation increased significantly.Furthermore, the expressions of phosphorylated STAT1 (p-STAT1) and phosphorylated JAK2 (p-JAK2) of DC-SOCS1 were much lower than those of the mDC, with statistically significant difference (t=-3.840 and 0.254, respectively, both P<0.01).Pathological analysis revealed that there existed moderate hepatic cells necrosis and less immune cell infiltration in DC-SOCS1 group accompanied with higher regulatory T lymphocytes proportion than those in ALF group and DC-VNG group.Survival rate of ALF with DC-SOCS1 treatment group was significantly higher than that of ALF group with statistical difference (x2=12.87, P<0.05).Conclusions DC-SOCS1could sustain an immature state and exhibit as regulatory DC through negative regulation of JAK2/STAT1 pathway with overexpression of SOCS1.Infusion of DC-SOCS1 could ameliorate ALF by inhibiting aggressive inflammation response with increased proportion of regulatory T cells in mice, which shows good therapeutic effect for ALF mice.