[Objective] This study aimed to obtain IL-10 (interleukin 10) full-length cDNA of common carp (Cyprinus carpio L.) and conduct the sequence analysis. [Method] The differentially expressed cDNA fragment was obtained by...[Objective] This study aimed to obtain IL-10 (interleukin 10) full-length cDNA of common carp (Cyprinus carpio L.) and conduct the sequence analysis. [Method] The differentially expressed cDNA fragment was obtained by DD-RTPCR (differential display RT-PCR). The cDNA library of peripheral blood leukocytes which were separated from common carp and stimulated by mitogen was screened with a probe labeled with DIG (digoxigenin). The IL-10 full-length cDNA was cloned from 0.8×104 pfu of recombinant phages, and the sequence analysis and homology comparison were carried out. [Result] Sequence analysis indicated that the IL-10 full-length cDNA of common carp was 1 117 bp long, containing a 55 bp 5’-UTR, a 522 bp 3’-UTR, and a 540 bp open reading frame(ORF) encoding 179 amino acids. In addition, there were three mRNA instability motifs (ATTTA) in the 3’-untranslated region. The deduced protein sequence shared typical sequence features of the IL-10 family. Homology comparison indicated that the obtained sequence shared 89.1% homology with the carp IL-10 gene from GenBank. [Conclusion] This study laid foundation for further study of the expression manner, functional characteristic and regulation mechanism of IL-10 in vivo and the interaction mechanism in the inflammatory reaction and immune response.展开更多
OBJECTIVES: To evaluate the behavior of serum interleukin 10 (IL-10) in patients with acute pancreatitis and to explore the relationship between this cytokine and the severity of the disease. METHODS: In 45 patients w...OBJECTIVES: To evaluate the behavior of serum interleukin 10 (IL-10) in patients with acute pancreatitis and to explore the relationship between this cytokine and the severity of the disease. METHODS: In 45 patients with acute pancreatitis, the serum concentrations of IL-10 was determined on days 1, 2, 3, 4, 5 after admission. Twelve healthy subjects were also studied as controls. These subjects were tested using a commercial ELISA kit. The severity of pancreatitis was determined according to APACHE Ⅱ score and Balthazar CT criteria. RESULTS: Healthy subjects had no detectable serum levels of IL-10. In acute pancreatitis patients, the serum IL-10 levels were increased on the first day after the onset of the disease and then progressively decreased in the following days. On the first day after the onset of acute pancreatitis, the serum levels of IL-10 in patients with mild acute pancreatitis were significantly higher than in those with severe acute pancreatitis. In the following days, however, no statistically significant difference was observed between the two groups. CONCLUSIONS: Serum IL-10 concentration reflects the severity of acute pancreatitis. IL-10 is a useful variable for early prediction of the prognosis of acute pancreatitis. The low values of serum IL-10 in patients with severe acute pancreatitis suggests that there may be altered down-regulation of immune system response. An enhanced release of IL-10 may be a method for early treatment of acute pancreatitis.展开更多
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 ra...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.展开更多
AIM: To investigate the effect of Clostridium difficile (C. difficile) infection in an interleukin 10-deficient (IL-10<sup>-/-</sup>) mouse model of inflammatory bowel disease.
BACKGROUND Interleukin 10 receptor alpha subunit(IL10RA)dysfunction is the main cause of very early-onset inflammatory bowel disease(VEO-IBD)in East Asians.AIM To identify disease-causing gene mutations in four patien...BACKGROUND Interleukin 10 receptor alpha subunit(IL10RA)dysfunction is the main cause of very early-onset inflammatory bowel disease(VEO-IBD)in East Asians.AIM To identify disease-causing gene mutations in four patients with VEO-IBD and verify functional changes related to the disease-causing mutations.METHODS From May 2016 to September 2020,four young patients with clinically diagnosed VEO-IBD were recruited.Before hospitalization,using targeted gene panel sequencing and trio-whole-exome sequencing(WES),three patients were found to harbor a IL10RA mutation(c.301C>T,p.R101W in one patient;c.537G>A,p.T179T in two patients),but WES results of the fourth patient were not conclusive.We performed whole-genome sequencing(WGS)on patients A and B and reanalyzed the data from patients C and D.Peripheral blood mononuclear cells(PBMCs)from patient D were isolated and stimulated with lipopolysaccharide(LPS),interleukin 10(IL-10),and LPS+IL-10.Serum IL-10 levels in four patients and tumor necrosis factor-α(TNF-α)in the cell supernatant were determined by enzyme-linked immunosorbent assay.Phosphorylation of signal transducer and activator of transcription 3(STAT3)at Tyr705 and Ser727 in PBMCs was determined by western blot analysis.RESULTS The four children in our study consisted of two males and two females.The age at disease onset ranged from 18 d to 9 mo.After hospitalization,a novel 333-bp deletion encompassing exon 1 of IL10RA was found in patients A and B using WGS and was found in patients C and D after reanalysis of their WES data.Patient D was homozygous for the 333 bp deletion.All four patients had elevated serum IL-10 levels.In vitro,IL-10-stimulated PBMCs from patient D failed to induce STAT3 phosphorylation at Tyr705 and only minimally suppressed TNF-αproduction induced by LPS.Phosphorylation at Ser727 in PBMCs was not affected by LPS or LPS+IL-10 in both healthy subjects and in patient D.CONCLUSION WGS revealed a novel 333-bp deletion of IL10RA in four patients with VEO-IBD,whereas the WES results were inconclusive.展开更多
Trypanosomosis is major drawback to profitable livestock production in sub-Sahara African, including Nigeria. Knowledge of the cytokines production in the phase of natural infection may help to better diagnose, treat ...Trypanosomosis is major drawback to profitable livestock production in sub-Sahara African, including Nigeria. Knowledge of the cytokines production in the phase of natural infection may help to better diagnose, treat and prevent bovine trypanosomosis. The purpose of the this study was to determine the levels of interferon-gamma (IFN-γ), interleukin-10 (IL-10) and cardiac troponin–I (cTnI) in the sera of cattle naturally infected with T. brucei, T. congolense and T. vivax and correlate these levels with parasitaemia and PCV of the infected animals. Five milliliter of blood samples were collected via the jugular vein from 411 randomly selected cattle into EDTA and non-citrated bottle. PCV was determined manually using HCT. Trypansomes were detected and characterized by microscopy and PCR, respectively. Serum levels of IFN-γ, IL-10 and cTnI were determined using commercial ELISA kit. Data were summarized using descriptive statistic and significance of differences determined by ANOVA. Of the 62 samples positive for trypanosomes by microscopy, 50 samples were confirmed to species level by PCR. The sera levels of IFN-γ, IL-10 and cTnI of infected cattle were higher than non-infected cattle. The differences were not significant (p γ, IL-10 and cTnI in cattle with natural trypanosomosis. Further investigation is required to understand the specific effect of trypanosomes on myocardiac integrity and interaction between the two cytokines in natural trypanosomosis in cattle.展开更多
文摘[Objective] This study aimed to obtain IL-10 (interleukin 10) full-length cDNA of common carp (Cyprinus carpio L.) and conduct the sequence analysis. [Method] The differentially expressed cDNA fragment was obtained by DD-RTPCR (differential display RT-PCR). The cDNA library of peripheral blood leukocytes which were separated from common carp and stimulated by mitogen was screened with a probe labeled with DIG (digoxigenin). The IL-10 full-length cDNA was cloned from 0.8×104 pfu of recombinant phages, and the sequence analysis and homology comparison were carried out. [Result] Sequence analysis indicated that the IL-10 full-length cDNA of common carp was 1 117 bp long, containing a 55 bp 5’-UTR, a 522 bp 3’-UTR, and a 540 bp open reading frame(ORF) encoding 179 amino acids. In addition, there were three mRNA instability motifs (ATTTA) in the 3’-untranslated region. The deduced protein sequence shared typical sequence features of the IL-10 family. Homology comparison indicated that the obtained sequence shared 89.1% homology with the carp IL-10 gene from GenBank. [Conclusion] This study laid foundation for further study of the expression manner, functional characteristic and regulation mechanism of IL-10 in vivo and the interaction mechanism in the inflammatory reaction and immune response.
文摘OBJECTIVES: To evaluate the behavior of serum interleukin 10 (IL-10) in patients with acute pancreatitis and to explore the relationship between this cytokine and the severity of the disease. METHODS: In 45 patients with acute pancreatitis, the serum concentrations of IL-10 was determined on days 1, 2, 3, 4, 5 after admission. Twelve healthy subjects were also studied as controls. These subjects were tested using a commercial ELISA kit. The severity of pancreatitis was determined according to APACHE Ⅱ score and Balthazar CT criteria. RESULTS: Healthy subjects had no detectable serum levels of IL-10. In acute pancreatitis patients, the serum IL-10 levels were increased on the first day after the onset of the disease and then progressively decreased in the following days. On the first day after the onset of acute pancreatitis, the serum levels of IL-10 in patients with mild acute pancreatitis were significantly higher than in those with severe acute pancreatitis. In the following days, however, no statistically significant difference was observed between the two groups. CONCLUSIONS: Serum IL-10 concentration reflects the severity of acute pancreatitis. IL-10 is a useful variable for early prediction of the prognosis of acute pancreatitis. The low values of serum IL-10 in patients with severe acute pancreatitis suggests that there may be altered down-regulation of immune system response. An enhanced release of IL-10 may be a method for early treatment of acute pancreatitis.
基金This research was supported by the National Natural Sciences Foundation (No.39730220).
文摘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.
文摘AIM: To investigate the effect of Clostridium difficile (C. difficile) infection in an interleukin 10-deficient (IL-10<sup>-/-</sup>) mouse model of inflammatory bowel disease.
基金the National Natural Science Foundation of China,No.81741103.
文摘BACKGROUND Interleukin 10 receptor alpha subunit(IL10RA)dysfunction is the main cause of very early-onset inflammatory bowel disease(VEO-IBD)in East Asians.AIM To identify disease-causing gene mutations in four patients with VEO-IBD and verify functional changes related to the disease-causing mutations.METHODS From May 2016 to September 2020,four young patients with clinically diagnosed VEO-IBD were recruited.Before hospitalization,using targeted gene panel sequencing and trio-whole-exome sequencing(WES),three patients were found to harbor a IL10RA mutation(c.301C>T,p.R101W in one patient;c.537G>A,p.T179T in two patients),but WES results of the fourth patient were not conclusive.We performed whole-genome sequencing(WGS)on patients A and B and reanalyzed the data from patients C and D.Peripheral blood mononuclear cells(PBMCs)from patient D were isolated and stimulated with lipopolysaccharide(LPS),interleukin 10(IL-10),and LPS+IL-10.Serum IL-10 levels in four patients and tumor necrosis factor-α(TNF-α)in the cell supernatant were determined by enzyme-linked immunosorbent assay.Phosphorylation of signal transducer and activator of transcription 3(STAT3)at Tyr705 and Ser727 in PBMCs was determined by western blot analysis.RESULTS The four children in our study consisted of two males and two females.The age at disease onset ranged from 18 d to 9 mo.After hospitalization,a novel 333-bp deletion encompassing exon 1 of IL10RA was found in patients A and B using WGS and was found in patients C and D after reanalysis of their WES data.Patient D was homozygous for the 333 bp deletion.All four patients had elevated serum IL-10 levels.In vitro,IL-10-stimulated PBMCs from patient D failed to induce STAT3 phosphorylation at Tyr705 and only minimally suppressed TNF-αproduction induced by LPS.Phosphorylation at Ser727 in PBMCs was not affected by LPS or LPS+IL-10 in both healthy subjects and in patient D.CONCLUSION WGS revealed a novel 333-bp deletion of IL10RA in four patients with VEO-IBD,whereas the WES results were inconclusive.
文摘Trypanosomosis is major drawback to profitable livestock production in sub-Sahara African, including Nigeria. Knowledge of the cytokines production in the phase of natural infection may help to better diagnose, treat and prevent bovine trypanosomosis. The purpose of the this study was to determine the levels of interferon-gamma (IFN-γ), interleukin-10 (IL-10) and cardiac troponin–I (cTnI) in the sera of cattle naturally infected with T. brucei, T. congolense and T. vivax and correlate these levels with parasitaemia and PCV of the infected animals. Five milliliter of blood samples were collected via the jugular vein from 411 randomly selected cattle into EDTA and non-citrated bottle. PCV was determined manually using HCT. Trypansomes were detected and characterized by microscopy and PCR, respectively. Serum levels of IFN-γ, IL-10 and cTnI were determined using commercial ELISA kit. Data were summarized using descriptive statistic and significance of differences determined by ANOVA. Of the 62 samples positive for trypanosomes by microscopy, 50 samples were confirmed to species level by PCR. The sera levels of IFN-γ, IL-10 and cTnI of infected cattle were higher than non-infected cattle. The differences were not significant (p γ, IL-10 and cTnI in cattle with natural trypanosomosis. Further investigation is required to understand the specific effect of trypanosomes on myocardiac integrity and interaction between the two cytokines in natural trypanosomosis in cattle.
