特应性和非特应性哮喘患儿血清白细胞介素-10和血浆内皮素-1表达水平的检测及其临床意义

目的:观察特应性与非特应性哮喘患儿血清白细胞介素-10(IL-10)、血浆内皮素-1(ET-1)和IgE水平,探讨其临床意义。方法:60例哮喘患儿同时采用过敏原皮肤点刺试验和血清IgE测定确定特应性状态,并根据结果分为特应性哮喘组32例与非特应性哮喘组28例,30例正常健康儿作为对照组。采用放射免疫法检测哮喘患儿在发作期和缓解期及对照组儿童血浆ET-1和IgE的含量,应用ELISA双抗体夹心法检测各组血清IL-10的含量。结果:发作期哮喘患儿血清IL-10的表达明显低于对照组(P<0.01),ET-1水平明显高于对照组(P<0.05),发作期及缓解期哮喘患儿的IgE水平均明显高于对照组(P<0.01)。哮喘组发作期患儿ET-1和IgE均高于缓解期患儿,IL-10低于缓解期患儿(P<0.01)。特应性组发作期患儿IL-10水平较非特应性组降低(P<0.05),特应性组发作期和缓解期患儿ET-1、IgE水平较非特应性组均增高(P<0.05)。支气管哮喘患儿IL-10的含量与ET-1及IgE分别呈明显负相关(r=-0.592,r=-0.894,P<0.05),ET-1与IgE呈明显正相关(r=0.623,P<0.05)。结论:血清IL-10和ET-1可能参与了小儿哮喘的病理生理过程,且特应性哮喘患者较非特应性哮喘患者血清IL-10、ET-1和IgE水平改变程度更明显。

Serum Levels of Interleukin-2 and -10 in Patients with Early Syphilis

Objective: To investigate the roles of interleukin-2(IL-2) and interleukin-10 (IL-10) in pathogenesis ofearly syphilis. Methods: The serum levels of IL-2 and IL-10 in 48patients with early syphilis were detected by ABC-ELISA. Results: (1) The level of IL-2 in the patients withearly syphilis was significantly higher than that inhealthy controls, while that of IL-10 was lower(P<0.001 and P<0.001). (2) The levels of IL-2 and IL-10 were almost identical in patients with primary andsecondary syphilis (P>0.05), as well as between dif-ferent RPR titers (P>0.05). (3) After therapy, the levelof IL-2 decreased markedly (P<0.05), while that of IL-10 increase (p>0.05). (4) A significant correlation wasfound between the serum levels of IL-2 and IL-10 (r=0.5385 P<0.05). Conclusions: Th1 up-regulation occurs in patientswith early syphilis, and plays an active role in fightingagainst TP infection.

Effects of interleukin-10 on activation and apoptosis of hepatic stellate cells in fibrotic rat liver

AIM： TO study the effects of interleukin-10 （IL-10） on the expression of o-smooth muscle actin （α-SMA）, nuclear factor-κB（NF-κB） and Fas/Fas ligand （FasL） in hepatic stellate cells of experimental rats with hepatic fibrosis. METHODS： Sixty clean SD rats were randomly divided into control group （group N）, liver fibrotic group （group C） and IL-10 treatment group （group I）. Control group received intraperitoneal injection of saline （2ml·kg^-1）, twice a week. Fibrotic group was injected intraperitoneally with 50% carbon tetrachloride （CCh） （2 ml·kg^-1）, twice a week. IL-10 treatment group was given IL-10 at a dose of 4 pg·kg^-1 20 minutes before CCl4 administration from the third week. Hepatic stellate cells （HSCs） were isolated from these rats at the seventh and eleventh weeks during the course of liver fibrosis, respectively. The expression of α-SMA and NF-κB in HSCs was measured by S-P immunohistochemistry. The expression of Fas and FasL mRNA was measured by RT-PCR. Furthermore, liver tissues were harvested from three groups at the same time. RESULTS： The CCh- induced experimental rat hepatic fibrosis model was established successfully. The purity of extracted hepatic stellate cells was about 95% and the yield of hepatic stellate cells was 1.2-2.3×10^6/g liver tissue averagely. The positive expression of α-SMA and NF-κB was 36.5% and 28.5% respectively in group N. The positive levels of α-SMA and NF-κB were increased significantly in group C compared to group N （P〈0.01）. The positive signals decreased significantly （P〈0.05） in group I. In the 11^th week, the HSCs of group I became round with visible pyknotic nuclei. The expression of NF-κB in group C was significantly increased in a timedependentmanner （P〈0.01）, but there was no difference in the α-SMA expression （P〉0.05）. The mRNA of Fas and FasL in group C was significantly increased in a timedependent manner compared to that in control group. After treated with IL-10, the expression level of Fas and FasL was higher in group I than in group C. CONCLUSION： The positive expression of α-SMA and NF-κB in hepatic stellate cells is decreased by ectogenic IL-10 in liver fibrosis induced by CCh. The expression of Fas and FasL is increased in the course of liver fibrosis, and is further increased by IL-10. IL-10 could inhibit the activation of HSCs and cause apoptosis of activated HSCs.

Therapeutic effect of interleukin-10 on CCI_4-induced hepatic fibrosis in rats

AIM： To study the therapeutic effect of exogenous interleuldn-10 on CCl4-induced hepatic fibrosis in rats and its passible mechanisms. METHODS： Fourty-seven SD rats were randomly divided into control group （group N） and CCl4-induced hepatic fibrosis model group （group C）. After CCl4 was given for 9 wk, the model group was divided into three groups. Rats in group H were put to death immediately, rats in group T were treated with IL-10 for another three wk and then put to death, rats in group R recovered after three weeks and were then killed. The degree of hepatic fibrosis was measured by HE staining and histological activity index （HAI）. Histological activity index （HAI）, change of collagen types Ⅰ and Ⅲ were measured by Picrosirius staining. The expression of TNF-α, HHP-2 and TIMP-1 in liver tissue was measured by S-P immunohis tochemistry.RESULTS： CCl4- induced experimental rat hepatic fibrosis model was established successfully. The degree of hepatic fibrosis was markedly lower in group T than in groups H and R, and there was no difference between the two groups. The expression of collagen types I and III was significantly suppressed in group T and was slightly suppressed in groups H and R. The positive levels of TNF-α, HHP-2 and TIHP-1 in group H increased significantly compared to those in group N （P〈0.01）. The positive signals decreased significantly in groups T and R （P〈0.01）, but positive score was significantly lower in group T than in group R （P〈 0.01）. CONCLUS10N： Exogenous IL-10 can reverse CCl4-induced hepatic fibrosis in rats. IL-10 may exert its reversible effects on hepatic fibrosis by blocking CCl4-induced inflammation, inhibiting expression of HHP-2 and TIMP-1 and promoting resolution of collagen types Ⅰ and Ⅲ.

Interleukin-10 and chronic liver disease

Interleukin （IL）-10 is an important immunoregulatory cytokine produced by many cell populations. Numerous investigations suggest that IL-10 plays a major role in chronic liver diseases. IL-10 gene polymorphisms are possibly assodated with liver disease susceptibility or se-verity. Recombinant human IL-10 has been produced and is currently tested in clinical trials. These trials may give new insights into the immunobiology of IL-10 and suggest that the IL-10/IL-10 receptor system may become a new therapeutic target.

Polymorphisms of interleukin-10 promoter are not associated with prognosis of advanced gastric cancer

AIM: To evaluate the association between of the interleukin-10 (IL-10) promoter polymorphisms and survival of advanced gastric cancer (GC) patients. METHODS: The IL-10 (-1082, rs1800896; -819, rs1800871; and-592, rs1800896) genotypes in 234 patients with advanced gastric cancer and in 243 healthy controls were determined by polymerase chain reaction-restriction fragment length polymorphism assay. Odds ratios (OR) and 95% confidence intervals (CI) were calculated by unconditional logistic regression for the associations between IL-10 genotypes and the risk of GC. The Kaplan-Meier method with log-rank testing was used to evaluate the association between genotype and survival of the patients.RESULTS: The IL-10 -1082 G allele and GCC (-1082, -819 and -592) haplotype were associated with increased gastric cancer risks (OR 1.2, 95% CI 0.6-3.2, P = 0.007, for -1082 G allele, OR = 2.3, 95% CI, 1.2-4.1, P = 0.005, for GCC haplotype, respectively). However, none of the three IL-10 gene polymorphisms (-1082, -819 and -592) was correlated with gastric cancer survival (P > 0.05), and none of the genotypes of the three IL-10 sites was found as independent prognostic risk factors in the multivariate test. CONCLUSION: IL-10 gene promoter polymorphisms may not be associated with the prognosis of advanced gastric cancer.

Prognostic performance of interleukin-10 in patients with chest pain and mild to moderate coronary artery lesions an 8-year follow-up study

Background Interleukin （IL）-10, IL-6 and their ratio （IL-6/IL-10） play an important role in the risk of developing coronary artery disease, and may correlate with its outcomes. Few clinical trials have investigated the prognostic impact of these factors on long-term car- diovascular events in patients presented with chest pain. Methods A prospective study was performed on 566 patients admitted with chest pain and identified mild to moderate coronary artery lesions. 1L-10, IL-6 and IL-6/IL-10 were measured. Results A total of 511 patients com- pleted the follow-up. The median follow-up time was 74 months. Kaplan-Meier analysis demonstrated a clear increase of the incidence of major adverse cardiac events during the follow-up period in patients with below-median levels of IL-10 （P = 0.006） and above-median levels of IL-6/IL-10 （P = 0.012）. Multivariate Cox proportional hazards analysis indicated the IL-10 levels to be strong independent predictors after adjustment for underlying confounders. Conclusions Elevated IL-10 levels are associated with a more favorable long-term prognosis in patients with chest pain and mild to moderate coronary artery lesions. IL-10 could be used for early risk assessment of long-term prognosis.

Interleukin-10 -1082 promoter polymorphism is not associated with susceptibility to esophageal squamous cell carcinoma and gastric cardiac adenocarcinoma in a population of high-incidence region of north China

AIM: To investigate the possible association of G→A single nucleotide polymorphism (SNP) at the -1082 position of interleukin (IL)-10 promoter with susceptibility to esophageal squamous cell carcinoma (ESCC) and gastric cardiac adenocarcinoma (GCA) in a population of a high incidence region of North China.METHODS: IL-10-G1082A promoter SNP was genotyped by polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) in 355 cancer patients (203ESCC and 152 GCA) and 443 healthy controls.RESULTS: Smoking significantly increased the risk of ESCC and GCA development (the age and sex adjusted OR = 1.42and 2.64, 95%CI = 1.11-1.81 and 1.46-4.76, respectively).Similarly, family history of upper gastrointestinal cancer (UGIC) significantly increased the risk of developing ESCC and GCA (the age and sex adjusted OR = 1.44 and 3.10,95%CI = 1.18-1.75 and 1.94-4.97, respectively). The A/A, A/G and G/G genotype frequencies of IL-10-G1082A were 60.3%, 37.0% and 2.7% in healthy controls, 57.6%,39.9% and 2.5% in ESCC and 61.2%, 36.8% and 2.0% in GCA patients, respectively. The frequencies of A and G alleles were 78.8% and 21.2% in healthy controls, 77.6%and 22.4% in ESCC patients and 79.6%, 20.4% in GCA patients. The distribution of genotype and allelotype in ESCC and GCA patients was not significantly different from that in healthy controls (P＞0.05). Compared to the A/A genotype, the combination of A/G and G/G genotypes did not show a significant effect on the risk of developing ESCC and GCA; the adjusted odds ratio was 0.92 (95%CI = 0.76-1.11) in ESCC and 0.95 (95% CI = 0.61-1.46)in GCA, respectively. When stratified for smoking status and family history of UGIC, the combination of A/G and G/G genotypes also did not show any significant influence on the risk of ESCC and GCA development compared to A/A genotypes.CONCLUSION: IL-10-G1082A polymorphism might not be used as a stratification marker to predicate the risk of ESCC and GCA development in North China.

Polymorphisms in interleukin-10 gene according to mutations of NOD2/CARD15 gene and relation to phenotype in Spanish patients with Crohn's disease

AIM： To examine the contribution of interleukin-10 （IL-10） gene polymorphisms to Crohn＇s disease （CD） phenotype, and the possible genetic epistasis between IL-10 gene polymorphisms and CARD15/NOD2 gene mutations. METHODS： A cohort of 205 Spanish unrelated patients with Crohn＇s disease recruited from a single center was studied. All patients were rigorously phenotyped and followed-up for at least 3 years （mean time, 12.5 years）. The clinical phenotype was established prior to genotyping. RESULTS： The correlation of genotype-Vienna classification groups showed that the Ueocolonic location was significantly associated with the -1082G allele in the NOD2/CARD15 mutation-positive patients （RR = 1.52, 95%CI, 1.21 to 1.91,P= 0.008）. The multivariate analysis demonstrated that the IL-10 G14 microsatellite allele in the NOD2/CARD15 mutation positive patients was associated with two risk factors, history of appendectomy （RR = 2.15, 95%CI = 1.1-4.30, P= 0.001） and smoking habit at diagnosis （RR= 1.29, 95%CI= 1.04-4.3, P= 0.04）. CONCLUSION： In Spanish population from Madrid, in CD patients carrying at least one NOD2/CARD15 mutation, the -1082G allele is assodated with ileocolonic disease and the IL-IOG14 microsatellite allele is associated with previous history of appendectomy and smoking habit at diagnosis. These data provide further molecular evidence for a genetic basis of the clinical heterogeneity of CD.

Interleukin-10 gene polymorphisms and hepatocellular carcinoma susceptibility:A meta-analysis

AIM: To assess the association between Interleu-kin-10 (IL-10) gene IL-10-1082 (G/A), IL-10-592(C/A), IL-10-819 (T/C) polymorphisms and hepatocellular carcinoma (HCC) susceptibility.METHODS: Two investigators independently searched the Medline, Embase, China National Knowledge Infrastructure, and Chinese Biomedicine Database. Summary odds ratios (ORs) and 95% conf idence intervals (95% CIs) for IL-10 polymorphisms and HCC were cal-culated in a fixed-effects model (the Mantel-Haenszel method) and a random-effects model (the DerSimonian and Laird method) when appropriate. RESULTS: This meta analysis included seven eligiblestudies, which included 1012 HCC cases and 2308 controls. Overall, IL-10-1082 G/A polymorphism was not associated with the risk of HCC (AA vs AG + GG, OR = 1.11, 95% CI = 0.90-1.37). When stratifying for ethnicity, the results were similar (Asian, OR = 1.12, 95% CI = 0.87-1.44; non-Asian, OR = 1.10, 95% CI = 0.75-1.60). In the overall analysis, the IL-10 polymorphism at position -592 (C/A) was identified as a genetic risk factor for HCC among Asians; patients carrying the IL-10-592*C allele had an increased risk of HCC (OR = 1.29, 95% CI = 1.12-1.49). No association was observed between the IL-10-819 T/C polymorphism and HCC susceptibility (TT vs TC + CC, OR = 1.02, 95% CI = 0.79-1.32).CONCLUSION: This meta-analysis suggests that IL-10-592 A/C polymorphism may be associated with HCC among Asians. IL-10-1082 G/A and IL-10-819 T/C polymorphisms were not detected to be related to the risk for HCC.

Association of Interleukin-10 Gene Haplotype with Gastric Cancer in a Chinese Population

OBJECTIVE Interleukin-10 （IL-10） is a multifunctional cytokine with both immunosuppressive and antiangiogenic functions.Previous studies have reported that IL-10 levels are signifi cantly elevated in patients with gastric cancer （GC）. It has also been confirmed that interindividual variations in IL-10 production are genetically attributed to the polymorphisms of IL-10 gene.Therefore, this study was designed to investigate whether the polymorphisms of IL-10 gene were associated with susceptibility to GC in the Chinese population.METHODS The serum levels of IL-10 were measured by radioimmunoassay. The single nucleotide polymorphisms （SNPs） at positions -1082A/G, -819T/C and -592A/C in the IL-10 gene promoter were analyzed using polymerase chain reactionrestriction fragment length polymorphism （PCR-RFLP）.RESULTS 220 patients with gastric cancer and 180 healthy controls were included in this study. The serum levels of IL-10 were signifi cantly higher in GC patients than healthy controls （Z = -19.13, P 〈 0.001）. Single SNP analysis showed that the -1082G allele, -1082AG and -819CC genotypes significantly increased in patients with GC （P = 0.029, 0.021, 0.039 respectively）. In a logistic regression analysis adjusted for age and sex, the -1082AG genotype was associated with an odds ratio of 1.974 （95% CI,1.14-3.391; P = 0.014）, and the -819CC genotype with an odds ratio of 2.496 （95% CI, 1.222-5.102; P = 0.012） for GC. Furthermore,haplotype analysis revealed that at least five haplotypes （ATA,ACC, GCC, ACA and ATC） were existent in this population.Also that the GCC haplotype was associated with a signifi cantly increased risk of GC as compared with the ATA haplotype （OR =1.90; 95% CI, 1.11-3.27; P = 0.02）.CONCLUSION The results indicate that the gene haplotype of IL-10 may contribute to the susceptibility to GC in the Chinese population.

Effect of 5-FU on modulation of disarrangement of immune-associated cytokines in experimental acute pancreatitis

AIM： To investigate the effects of 5-Fluorouracil （5-FU） on modulation of pro-inflammatory and anti-inflammatory cytokines in acute pancreatitis and the mechanism of it in the treatment of acute pancreatitis. METHODS： Male Sprague Dawley rats were assigned to 3 Groups： Group A, sham operated rats as controls （n = 7）; Group B, acute pancreatitis induced by ductal injection with 5% sodium cholate at a volume of 1.0 mL/kg without any other treatment; Group C, after the pancreatitis was induced as in Group B, the rats were injected intravenously with 5-FU 40 mg/kg. The animals in Groups B and C were killed at 2, 6 and 24 h after operation （n = 7）, and blood samples were taken for measurement of tumor necrosis factor-α （TNF-α）, interleukin-1 （IL-1）, interleukin-6 （IL-6） （by bioassay）, and interleukin-10 （IL-10）, transforming growth factor-β （TGF-β） （by ELISA）. The wet weight of pancreatic tissue, serum amylase levels and white blood cells were also measured. RESULTS： Four rats in Group B and one in Group C died after pancreatitis was induced. Both pro-inflammatory cytokines （TNF-α, IL-1, IL-6） at the 2 and 6 h period and the anti-inflammatory cytokines （IL-10, TGF-β） at 24 h increased significantly （P 〈 0.05） in rats of Group B. After treatment with 5-FU, TNF-α, IL-1, and IL-6 in serum of rats of Group C were inhibited at 2 and 6 h after operation （P 〈 0.05）, and IL-IO, TGF-13 were inhibited at 24 h compared to Group B （P 〈 0.05）. Obvious improvements in the severity of the acute pancreatitis, including the amylase levels, wet weight of pancreatic tissue and neutrophil counts, were also observed after treatment with 5-FU. CONCLUSION： 5-FU is an anti-metabolic and immunosuppressive agent which can minimize the abnormal immune o/tokine response and relieve the pathophysiological disorders associated with experimental acute pancreatitis.

INTERLEUKIN 10 INHIBITS THE RAT VSMC PROLIFERATION AND COLLAGEN SECRETION STIMULATED BY ANGIOTENSIN II

Objective. To study the effect of interleukin 10 (IL- 10) on the angiotensin II (AngII) stimulated rat VSMC proliferation and collagen secretion, and furthermore, explore its mechanism. Methods. On cultured VSMC of rat, 3H- thymine (3H- TdR) and 3H- proline incorporations were used to evaluate the DNA and collagen synthesis, respectively. Western blot and immunoprecipitation were applied to assay the expression and activity of focal adhesion kinase (FAK), respectively. Results. IL- 10 (10－ 8～ 10－ 10g/ml) inhibited the increase of 3H- TdR and 3H- proline incorporation as well as FAK activity, which was induced by 10－ 7mol/L AngII (P< 0.05 or P< 0.01). IL- 10 also obviously downregulated the synthesis and secretion of collagen by AngII stimulated VSMC. But there was no difference in the protein expression of FAK among all the groups (P >0.05). Conclusion. IL- 10 antagonizes the VSMC proliferation and collagen synthesis by regulating FAK activity stimulated by AngII.

Intravenous injection of mesenchymal stem cells is effective in treating liver fibrosis

AIM: To compare the influence of different transplant sites in bone marrow mesenchymal stem cell (MSC)-based therapy for liver fibrosis. METHODS: MSCs isolated from Sprague Dawley (SD) rats were induced into hepatocyte-like cells. Liver fibrosis in SD rats was induced with carbon tetrachloride. Following hepatocyte induction in vitro, 4',6-diamidino- 2-phenylindole (DAPI)-labeled MSCs were transplanted by intravenous, intrahepatic, and intraperitoneal injection. Histopathological staining, immunohistochemistry, and biochemical analysis were used to compare the morphological and functional liver regeneration among different MSC injection modalities. The expression differences of interleukins, growth factor, extracellular matrix, matrix metalloproteinases, and tissue inhibitor of metalloproteinase were examined by real-time reverse transcription-polymerase chain reaction (RT-PCR) andenzyme linked immunosorbent assay (ELISA). RESULTS: Four days after exposure to hepatocyte differentiation medium, MSCs that did not express hepatocyte markers could express α-fetoprotein, albumin, and cytokeratin 18. The results of histopathological staining, immunohistochemistry, and biochemical analysis indicated that intravenous injection is more effective at rescuing liver failure than other injection modalities. DAPI-labeled cells were found around liver lobules in all three injection site groups, but the intravenous group had the highest number of cells. PCR and ELISA analysis indicated that interleukin-10 (IL-10) was highest in the intravenous group, whereas il1β, il6, tnfα and tgfβ, which can be regulated by IL10 and are promoters of liver fibrosis, were significantly lower than in the other groups. CONCLUSION: MSC administration is able to protect against liver fibrosis. Intravenous injection is the most favorable treatment modality through promotion of IL10 expression.

Effect of Jiaweiwumei decoction on regulatory T cells and interleukin-10 in a rat model of ulcerative colitis

OBJECTIVE:To investigate the effect of a decoction made from the Traditional Chinese Medicine Wumei pill,on regulatory T cells and interleukin-10(IL-10) in a rat model of ulcerative colitis induced by2,4,6-trinitrobenzene sulfonic acid(TNBS).METHODS:Rat ulcerative colitis was induced with TNBS.All modeled rats were randomly divided into six groups:normal control group;model group;sulfasalazine suppositories treatment group;and high,moderate,and low dosage of Jiaweiwumei decoction groups(12 rats each).Colon injury index was evaluated after 14 days.After peripheral blood lymphocyte separation,CD4 + T cells and CD4+/CD25+ T cell percentage was detected by flow cytometry.The content of IL-10 in serum and intestinal mucosa tissue was detected by sandwich enzyme-linked immunosorbent assay.RESULTS:Colon injury indices in the decoction groups were effectively reduced,compared with the model group(P < 0.05).Compared with that of the control group,the CD4+/CD25+ to CD4+ T lymphocyte ratio of the model group was significantly lower,while the decoction treatment improved the CD4 +/CD25 + to CD4 + T lymphocyte ratio(P <0.05).The serum and mucosal IL-10 content of the model group was significantly lower(P < 0.05) than that in the control group,while the decoction group had significantly higher serum and intestinal mucosal IL-10 content than that in the model group(P < 0.05).The regulatory T cell content was negatively correlated with the colonic injury index(r = 0.68,P < 0.05),and positively correlated with the content of serum IL-10(r = 0.87,P < 0.05) and intestinal mucosal IL-10(r= 0.79,P < 0.05).CONCLUSION:Jiaweiwumei decoction had significant effects on regulatory T cells and IL-10 in rats with TNBS-induced ulcerative colitis.

Pharmacological approach to acute pancreatitis

The aim of the present review is to summarize the current knowledge regarding pharmacological prevention and treatment of acute pancreatitis (AP) based on experimental animal models and clinical trials. Somatostatin (SS) and octreotide inhibit the exocrine production of pancreatic enzymes and may be useful as prophylaxis against Post Endoscopic retrograde cholangiopancreatography Pancreatitis (PEP). The protease inhibitor Gabexate mesilate (GM) is used routinely as treatment to AP in some countries, but randomized clinical trials and a meta-analysis do not support this practice. Nitroglycerin (NGL) is a nitrogen oxide (NO) donor, which relaxes the sphincter of Oddi. Studies show conflicting results when applied prior to ERCP and a large multicenter randomized study is warranted. Steroids administered as prophylaxis against PEP has been validated without effect in several randomized trials. The non-steroidal anti-inflammatory drugs (NSAID) indomethacin and diclofenac have in randomized studies showed potential as prophylaxis against PEP. Interleukin 10 (IL-10) is a cytokine with anti-inflammatory properties but two trials testing IL-10 as prophylaxis to PEP have returned conflicting results. Antibodies against tumor necrosis factor-alpha (TNF-α) have a potential as rescue therapy but no clinical trials are currently being conducted. The antibiotics beta- lactams and quinolones reduce mortality when necrosis is present in pancreas and may also reduce incidence of infected necrosis. Evidence based pharmacological treatment of AP is limited and studies on the effect of potent anti-inflammatory drugs are warranted.

Direct protective effect of interleukin-10 on articular chondrocytes in vitro

Abstract:Objective To assess whether interleukin-10 (IL-10) is chondroprotective in vitro. Methods Chondrocytes were isolated from femoral cartilage of rats (7-10 days) by digestion with collagenase Ⅱ. The first passage cells were grown in 24- well plates with DMEM, supplemented with 10% fetal bovine serum, for 2-4 days. The cells were then cultured in 0.1% fetal bovine serum DMEM medium, and given respectively interleukin-1 (IL-1) 100?μ/ml, IL-1 100?μ/ml+recombinant murine interleukin-10 (rmIL-10) 20?ng/ml, rmIL-10 20?ng/ml, and cultured for 48 hours. Scanning electron morphology and immunohistochemical study of nitric oxide synthase 2 and matric metalloproteinase 3 mRNA in situ hybridization were performed. Cell proliferation and morphology were observed under inverted microscope from the beginning of cell culture for three weeks. Results IL-1 stimulated granule production in the cytoplasma of chondrocytes, and the cells died in the second and third weeks of culture. IL-10 antagonized IL-1, protected the cells from death and maintained chondrocyte proliferation. Scanning electron morphology showed that IL-1 stimulated the formation of numerous microvilli on the cell surface, while thin and less numerous microvilli were found in cultures with IL-10. Immunohistochemical study and in situ hybridization showed that IL-10 inhibited NOS2 and MMP3 expression.Conclusion IL-10 not only inhibits the synthesis of inflammatory cytokines, but also directly protects chondrocytes by antagonizing IL-1.

Octreotide ameliorates gastric lesions in chronically mild stressed rats

AIM:To evaluate the effect of chronic mild stress(CMS) on the emergence of gastric ulcers and possible modulation by octreotide,a synthetic somatostatin analogue. METHODS:Adult male Wistar rats were subjected to nine different unpredictable random stress procedures for 21 d,a multifactorial interactional animal model for CMS.Octreotide was administered daily for 21 d at two dose levels(50 and 90μg/kg)before exposure to stress procedure.Macro-and microscopical assessments were made,in addition to quantification of plasma corticosterone and gastric mucosal inflammatory,oxidative stress, and apoptotic biomarkers. RESULTS:Exposure to CMS elevated plasma corticosterone(28.3±0.6μg/dL,P=0.002),an event that was accompanied by gastric lesions(6.4±0.16 mm,P=0.01) and confirmed histopathologically.Moreover,the insult elevated gastric mucosal lipid peroxides(13±0.5 nmol/g tissue,P=0.001),tumor necrosis factor-α(3008.6±78.18 pg/g tissue,P<0.001),prostaglandin E2(117.1 ±4.31 pg/g tissue,P=0.002),and caspase-3 activity (2.4±0.14 OD/mg protein,P=0.002).Conversely,CMS mitigated interleukin-10(627.9±12.82 pg/g tissue,P= 0.001).Furthermore,in animals exposed to CMS,octreotide restored plasma corticosterone(61%and 71%from CMS,P=0.002)at both dose levels.These beneficial effects were associated with a remarkable suppression of gastric lesions(38%and 9%from CMS,P=0.01)and reversal of derangements in gastric mucosa. CONCLUSION:The current investigation provides evidence that exposure to CMS induces gastric ulceration, which was alleviated by administration of octreotide possibly possessing antioxidant,anti-inflammatory,and anti-apoptotic actions.

Selective decrease in colonic CD56^+ T and CD161^+ T cells in the inflamed mucosa of patients with ulcerative colitis

AIM： To investigate the role of local colonic mucosal NK receptor-positive T （NKR＋ T） cells in the regulation of intestinal inflammation, we analyzed the population and function of these cells in ulcerative colitis （UC）. METHODS： Colonic mucosal tissues were obtained from colonoscopic biopsies of the descending colon from 96 patients with UC （51 endoscopically uninflamed, 45 inflamed） and 18 normal controls. Endoscopic appearance and histologic score at the biopsied site were determined by MaLts＇ classification. A single cell suspension was prepared from each biopsy by collagenase digestion. Two NKR^＋ T cell subsets, CD56^＋ （CD56^＋CD3^＋） T cells and CD161＋ （CD161^＋CD3^＋） T cells, were detected by flow cytometric analysis. Intracellular cytokine analysis for anti-inflammatory cytokine interleukin-10 （IL-10） was performed by in vitro stimulation with phorbol-myristateacetate （PMA） and ionomycin. RESULTS： CD56^＋ T cells and CD161^＋ T cells are present in the normal human colon and account for 6.7% and 21.3% of all mononuclear cells, respectively. The populations of both CD56＋ T cells and CD161^＋ T cells were decreased significantly in the inflamed mucosa of UC. In contrast, the frequency of conventional T cells （CD56 CD3^＋ cells and CD161CD3^＋ cells） was similar among the patient and control groups. The populations of NKR^＋ T cells were correlated inversely with the severity of inflammation, which was classified according to the endoscopic and histologic Marts＇ criteria. Interestingly, approximately 4% of mucosal NKR＋ T cells expressing IL-10 were detected by in vitro stimulation with PMA and ionomycin.CONCLUSION： Selective reduction in the population of colonic mucosal NKR＋T cells may contribute to the development of intestinal inflammation in UC.

Hepatitis C virus and ethanol alter antigen presentation in liver cells

Alcoholic patients have a high incidence of hepatitis C virus （HCV） infection. Alcohol consumption enhances the severity of the HCV disease course and worsens the outcome of chronic hepatitis C. The accumulation of virally infected cells in the liver is related to the HCV- induced inability of the immune system to recognize infected cells and to develop the immune responses. This review covers the effects of HCV proteins and ethanol on major histocompatibility complex （MHC） class Ⅰ- and class Ⅱ-restricted antigen presentation. Here, we discuss the liver which functions as an immune privilege organ; factors, which affect cleavage and loading of antigenic peptides onto MHC class I and class ~I in hepatocytes and dendritic cells, and the modulating effects of ethanol and HCV on antigen presentation by liver cells. Altered antigen presentation in the liver limits the ability ＇of the immune system to clear HCV and infected cells and contributes to disease progression. HCV by itself affects dendritic cell function, switching their cytokine profile to the suppressive phenotype of interleukin-10 （IL-10） and transforming growth factor beta （TGFβ） predominance, preventing cell maturation and allostimulation capacity. The synergistic action of ethanol with HCV results in the suppression of MHC class Ⅱ-restricted antigen presentation. In addition, ethanol metabolism and HCV proteins reduce proteasome function and interferon signaling, thereby suppressing the generation of peptides for MHC class I -restricted antigen presentation. Collectively, ethanol exposure further impairs antigen presentation in HCV-infected liver cells, which may provide a partial explanation for exacerbations and the poor outcome of HCV infection in alcoholics.