BACKGROUND Defective neutrophil regulation in inflammatory bowel disease(IBD)is thought to play an important role in the onset or manifestation of IBD,as it could lead to damage of the intestinal mucosal barrier by th...BACKGROUND Defective neutrophil regulation in inflammatory bowel disease(IBD)is thought to play an important role in the onset or manifestation of IBD,as it could lead to damage of the intestinal mucosal barrier by the infiltration of neutrophils in the inflamed mucosa and the accumulation of pathogens.Like neutrophils in the context of innate immune responses,immunoglobulin A(IgA)as an acquired immune response partakes in the defense of the intestinal epithelium.Under normal conditions,IgA contributes to the elimination of microbes,but in connection with the loss of tolerance to chitinase 3-like 1(CHI3L1)in IBD,IgA could participate in CHI3L1-mediated improved adhesion and invasion of potentially pathogenic microorganisms.The tolerance brake to CHI3L1 and the occurrence of IgA autoantibodies to this particular target,the exact role and underlying mechanisms of CHI3L1 in the pathogenesis of IBD are still unclear.AIM To determine the predictive potential of Ig subtypes of a novel serological marker,anti-CHI3L1 autoantibodies(aCHI3L1)in determining the disease phenotype,therapeutic strategy and long-term disease course in a prospective referral cohort of adult IBD patients.METHODS Sera of 257 Crohn’s disease(CD)and 180 ulcerative colitis(UC)patients from a tertiary IBD referral center of Hungary(Division of Gastroenterology,Department of Internal Medicine,Faculty of Medicine,University of Debrecen)were assayed for IgG,IgA,and secretory IgA(sIgA)type aCHI3L1 by enzyme-linked immunosorbent assay using recombinant CHI3L1,along with 86 healthy controls(HCONT).RESULTS The IgA type was more prevalent in CD than in UC(29.2%vs 11.1%)or HCONT(2.83%;P<0.0001 for both).However,sIgA subtype aCHI3L1 positivity was higher in both CD and UC patients than in HCONT(39.3%and 32.8%vs 4.65%,respectively;P<0.0001).The presence of both IgA and sIgA aCHI3L1 antibodies was associated with colonic involvement(P<0.0001 and P=0.038,respectively)in patients with CD.Complicated disease behavior at sample procurement was associated with aCHI3L1 sIgA positivity(57.1%vs 36.0%,P=0.009).IgA type aCH3L1 was more prevalent in patients with frequent relapse during the disease course in the CD group(46.9%vs 25.7%,P=0.005).In a group of patients with concomitant presence of pure inflammatory luminal disease and colon involvement at the time of diagnosis,positivity for IgA or sIgA type aCH3L1 predicted faster progression towards a complicated disease course in time-dependent models.This association disappeared after merging subgroups of different disease locations.CONCLUSION CHI3L1 is a novel neutrophil autoantigenic target in IBD.The consideration of antibody classes along with location-based prediction may transform the future of serology in IBD.展开更多
AIM: To assess blood chitinase 3-like 1(CHi3L1) levels for 2 mo after minimally invasive colorectal resection(MICR) for colorectal cancer(CRC). METHODS: CRC patients in an Institutional Review Board approved data/plas...AIM: To assess blood chitinase 3-like 1(CHi3L1) levels for 2 mo after minimally invasive colorectal resection(MICR) for colorectal cancer(CRC). METHODS: CRC patients in an Institutional Review Board approved data/plasma bank who underwent elective MICR for whom preoperative(PreO p), early postoperative(PostO p), and 1 or more late PostO p samples [postoperative day(POD) 7-27] available were included. Plasma CHi3L1 levels(ng/m L) were determined in duplicate by enzyme linked immunosorbent assay. RESULTS: PreOp and PostOp plasma sample were available for 80 MICR cancer patients for the study. The median PreOp CHi3L1 level was 56.8 CI: 41.9-78.6 ng/mL(n = 80). Significantly elevated(P < 0.001) median plasma levels(ng/mL) over PreOp levels were detected on POD1(667.7 CI: 495.7, 771.7; n = 79), POD 3(132.6 CI: 95.5, 173.7; n = 76), POD7-13(96.4 CI: 67.7, 136.9; n = 62), POD14-20(101.4 CI: 80.7, 287.4; n = 22), and POD 21-27(98.1 CI: 66.8, 137.4; n = 20, P = 0.001). No significant difference in plasma levels were noted on POD27-41. CONCLUSION: Plasma CHi3L1 levels were significantly elevated for one month after MICR. Persistently elevated plasma CHi3L1 may support the growth of residual tumor and metastasis.展开更多
Background:Serum chitinase-3-like protein 1(CHI3L1)is a potential biomarker for fibrosis assessment.We aimed to evaluate serum CHI3L1 as a noninvasive diagnostic marker for chronic hepatitis B virusrelated fibrosis.Me...Background:Serum chitinase-3-like protein 1(CHI3L1)is a potential biomarker for fibrosis assessment.We aimed to evaluate serum CHI3L1 as a noninvasive diagnostic marker for chronic hepatitis B virusrelated fibrosis.Methods:Serum CHI3L1 levels were measured by ELISA in 134 chronic hepatitis B(CHB)patients.Significant fibrosis was defined as a liver stiffness>9.7 kPa.The performance of CHI3L1 was assessed and compared to that of other noninvasive tests by receiver operating characteristic(ROC)analysis.Results:Serum CHI3L1 levels were significantly higher in CHB patients with significant hepatic fibrosis(≥F2)than in those without significant hepatic fibrosis(<F2)(56.5 ng/mL vs.81.9 ng/mL,P<0.001).In CHB patients,the specificity and sensitivity of CHI3L1 for predicting significant fibrosis were 75.6%and 59.1%,respectively,with a cut-off of 76.0 ng/mL and an area under the ROC curve of 0.728(95%CI:0.637–0.820).Conclusions:Serum CHI3L1 levels could be an effective new serological biomarker for the diagnosis of liver fibrosis.Moreover,CHI3L1 is feasible in monitoring disease progression.展开更多
AIM To identify whether chitinase 3-like 1(CHI3 L1) serves as a suitable biomarker for the prognosis of esophageal squamous cell carcinoma(ESCC) and to analyze this protein's cellular source.METHODS An ELISA was c...AIM To identify whether chitinase 3-like 1(CHI3 L1) serves as a suitable biomarker for the prognosis of esophageal squamous cell carcinoma(ESCC) and to analyze this protein's cellular source.METHODS An ELISA was conducted to detect the concentration of CHI3 L1 in the serum of 150 ESCC patients diagnosed between January 2001 and February 2005. The prognostic relevance of CHI3 L1 was evaluated by a Kaplan-Meier and Cox regression analysis. The immunohistochemistry was reanalyzed,and fluorescent staining was utilized to explore the cellular origins of CHI3 L1. We stimulated monocyte-derived macrophages(MDMs) with either IL-6 or the supernatant of the ESCC cell line Eca-109 and later investigated the level of CHI3 L1 by q PCR and ELISA.RESULTS The level of serum CHI3 L1 was higher in older patients(≥ 60) than in patients under the age of 60(P = 0.001). The patients with higher levels of CHI3 L1 had a significantly shorter overall survival,whereas the traditional markers,carcinoembryonic antigen and squamous cell carcinoma antigen,were less effective(P > 0.05). A multivariate Cox analysis(P = 0.001) indicated that CHI3 L1 was an independent prognostic factor for ESCC patients. Peritumoral macrophages in ESCC exhibited high levels of CHI3 L1. Interleukin-6(IL-6) and the supernatant of Eca-109 containing IL-6 stimulated MDMs to secrete CHI3 L1. The serum concentration of CHI3 L1 in the ESCC patients showed a weak correlation with the laboratory inflammatory parameters neutrophil(NEU,P = 0.045),neutrophil/lymphocyte rate(NLR,P = 0.016),and C-reactive protein(CRP,P < 0.001).CONCLUSION Our study first established a connection between the pretreated CHI3 L1 and patients with ESCC,and the serum CHI3 L1 was primarily secreted by ESCC-surrounded macrophages.展开更多
AIM:To evaluate the potential of two trabecular meshwork(TM)-specific promoters,Chitinase 3-like 1(Ch3L1)and matrix gla protein(MGP),for improving specificity and safety in glaucoma gene therapy based on self-compleme...AIM:To evaluate the potential of two trabecular meshwork(TM)-specific promoters,Chitinase 3-like 1(Ch3L1)and matrix gla protein(MGP),for improving specificity and safety in glaucoma gene therapy based on self-complementary AAV2(scAAV2)vector technologies.METHODS:An scAAV2 vector with C3 transferase(C3)as the reporter gene(scAAV2-C3)was selected.The scAAV2-C3 vectors were driven by Ch3L1(scAAV2-Ch3L1-C3),MGP(scAAV2-MGP-C3),enhanced MGP(scAAV2-eMGP-C3)and cytomegalovirus(scAAV2-CMV-C3),respectively.The cultured primary human TM cells were treated with each vector at different multiplicities of infections.Changes in cell morphology were observed by phase contrast microscopy.Actin stress fibers and Rho GTPases/Rho-associated protein kinase pathway-related molecules were assessed by immunofluorescence staining,real-time quantitative polymerase chain reaction and Western blot.Each vector was injected intracamerally into the one eye of each rat at low and high doses respectively.In vivo green fluorescence was visualized by a Micron III Retinal Imaging Microscope.Intraocular pressure(IOP)was monitored using a rebound tonometer.Ocular responses were evaluated by slit-lamp microscopy.Ocular histopathology analysis was examined by hematoxylin and eosin staining.RESULTS:In TM cell culture studies,the vectormediated C3 expression induced morphologic changes,disruption of actin cytoskeleton and reduction of fibronectin expression in TM cells by inhibiting the Rho GTPases/Rhoassociated protein kinase signaling pathway.At the same dose,these changes were significant in TM cells treated with scAAV2-CMV-C3 or scAAV2-Ch3L1-C3,but not in cells treated with scAAV2-eMGP-C3 or scAAV2-MGP-C3.At lowinjected dose,the IOP was significantly decreased in the scAAV2-Ch3L1-C3-injected eyes but not in scAAV2-MGPC3-injected and scAAV2-eMGP-C3-injected eyes.At highinjected dose,significant IOP reduction was observed in the scAAV2-eMGP-C3-injected eyes but not in scAAV2-MGP-C3-injected eyes.Similar to scAAV2-CMV-C3,scAAV2-Ch3L1-C3 vector showed efficient transduction both in the TM and corneal endothelium.In anterior segment tissues of scAAV2-eMGP-C3-injected eyes,no obvious morphological changes were found except for the TM.Inflammation was absent.CONCLUSION:In scAAV2-transduced TM cells,the promoter-driven efficiency of Ch3L1 is close to that of cytomegalovirus,but obviously higher than that of MGP.In the anterior chamber of rat eye,the transgene expression pattern of scAAV2 vector is presumably affected by MGP promoter,but not by Ch3L1 promoter.These findings would provide a useful reference for improvement of specificity and safety in glaucoma gene therapy using scAAV2 vector.展开更多
目的探讨外周血类黏蛋白1样蛋白3(ORMDL3)水平与过敏性哮喘(AA)患儿Th1/Th2平衡的关系。方法选取64例AA患儿为AA组,另选取48例同时期体检健康儿童为对照组;检测两组血清Ig E、嗜酸性粒细胞比率(EOSR),外周血ORMDL3 m RNA、Th1/Th2水平,...目的探讨外周血类黏蛋白1样蛋白3(ORMDL3)水平与过敏性哮喘(AA)患儿Th1/Th2平衡的关系。方法选取64例AA患儿为AA组,另选取48例同时期体检健康儿童为对照组;检测两组血清Ig E、嗜酸性粒细胞比率(EOSR),外周血ORMDL3 m RNA、Th1/Th2水平,以及呼出气一氧化氮(Fe NO)、达峰容积比(VPEF/VE)、达峰时间比(TPTEF/TE)。Pearson相关性分析AA组外周血ORMDL3 m RNA与Th1/Th2水平及两者与Ig E、EOSR、Fe NO、VPEF/VE、TPTEF/TE的相关性,多因素Logistics回归分析AA危险因素。结果AA组血清Ig E、EOSR、Fe NO和外周血ORMDL3 m RNA、Th2水平高于对照组,VPEF/VE、TPTEF/TE、Th2水平和Th1/Th2低于对照组(P均<0.05);AA患儿外周血ORMDL3 m RNA与Th1/Th2水平呈负相关(r=-7.103,P<0.05)。外周血ORMDL3 m RNA水平与Ig E、EOSR、Fe NO呈正相关,与VPEF/VE、TPTEF/TE呈负相关(P均<0.05),Th1/Th2水平与Ig E、EOSR、Fe NO呈负相关,与VPEF/VE、TPTEF/TE呈正相关(P均<0.05)。多因素Logistics回归分析显示,Ig E(OR=1.103,95%CI=1.019~1.193)和ORMDL3 m RNA(OR=13.697,95%CI=2.203~15.161)为AA发生独立危险因素,Th1/Th2(OR=0.040,95%CI=0.004~0.396)为保护因素(P均<0.05)。结论AA患儿外周血ORMDL3 m RNA水平明显升高,Th1/Th2处失衡状态,二者可作为评估AA病情的指标。展开更多
Sheath blight disease (ShB) has a severe impact on the production of rice. ABI3/VP1-like 1(RAVL1) negatively regulated the rice defense mechanism against ShB, however, this regulatorymechanism is not clearly understoo...Sheath blight disease (ShB) has a severe impact on the production of rice. ABI3/VP1-like 1(RAVL1) negatively regulated the rice defense mechanism against ShB, however, this regulatorymechanism is not clearly understood. In this study, we identified that indeterminate domain 3 (IDD3) waspositively regulated by RAVL1. Further, chromatin immunoprecipitation (ChIP) assay, yeast one-hybridassay and transient expression assay indicated a direct binding between RAVL1 and the IDD3 promoterregion. IDD3 was ubiquitously expressed in different tissues and at different stages, and its expressionwas significantly enhanced by Rhizoctonia solani infection. IDD3 exhibited transcription activation activityin yeast and IDD3-GFP was found to be localized in the nucleus. IDD3 mutants exhibited no significantdifferences in response to ShB, while IDD3 overexpressors were more susceptible to ShB compared withwild type (WT) plants. Furthermore, IDD3 repressors were less susceptible to R. solani than WT plants.Interestingly, the expression of brassinosteroid-related genes (D2, D11 and BRI1) was lower in IDD3repressors and higher in IDD3 overexpressors compared with WT. However, the ChIP assay revealedthat IDD3 did not directly bind to the D2 and D11 promoters. Overexpression of IDD3 in BRI1 mutantd61-1 inhibited the activity of IDD3, reducing its susceptibility to ShB compared with IDD3 overexpressorand WT plants, indicating that IDD3 negatively regulated the rice defense mechanism against ShB by activatingthe BR signaling pathway. Thus, our analyses provided information to enhance the understanding of therice defense mechanism against ShB.展开更多
Objective Current commercially available immunological tests cannot be used for discriminating active tuberculosis(TB)from latent TB infection.To evaluate the value of biomarker candidates in the diagnosis of active T...Objective Current commercially available immunological tests cannot be used for discriminating active tuberculosis(TB)from latent TB infection.To evaluate the value of biomarker candidates in the diagnosis of active TB,this study aimed to identify differentially expressed genes in peripheral blood mononuclear cells(PBMCs)between patients with active TB and individuals with latent TB infection by transcriptome sequencing.Methods The differentially expressed genes in unstimulated PBMCs and in Mycobacterium tuberculosis(Mtb)antigen-stimulated PBMCs from patients with active TB and individuals with latent TB infection were identified by transcriptome sequencing.Selected candidate genes were evaluated in cohorts consisting of 110 patients with TB,30 individuals with latent TB infections,and 50 healthy controls by quantitative real-time RT-PCR.Receiver operating characteristic(ROC)curve analysis was performed to calculate the diagnostic value of the biomarker candidates.Results Among the differentially expressed genes in PBMCs without Mtb antigen stimulation,interferon-induced protein with tetratricopeptide repeats 3(IFIT3)had the highest area under curve(AUC)value(0.918,95%CI:0.852-0.984,P<0.0001)in discriminating patients with active TB from individuals with latent TB infection,with a sensitivity of 91.86%and a specificity of 84.00%.In Mtb antigen-stimulated PBMCs,orosomucoid 1(ORM1)had a high AUC value(0.833,95%CI:0.752-0.915,P<0.0001),with a sensitivity of 81.94%and a specificity of 70.00%.Conclusion IFIT3 and ORM1 might be potential biomarkers for discriminating active TB from latent TB infection.展开更多
目的对2021年采集于我国福建的A组轮状病毒(group A rotavirus,RVA)G3P[8]毒株FJ21351116进行全基因组分子特征分析。方法使用高灵敏度A组轮状病毒全基因组测序方法对FJ21351116进行全基因组测序。用MEGA11.0、Geneious9.0.2和DNASTAR...目的对2021年采集于我国福建的A组轮状病毒(group A rotavirus,RVA)G3P[8]毒株FJ21351116进行全基因组分子特征分析。方法使用高灵敏度A组轮状病毒全基因组测序方法对FJ21351116进行全基因组测序。用MEGA11.0、Geneious9.0.2和DNASTAR软件通过核酸序列分析评估病毒的基因组特征。使用BioEdit v.7.0.9.0和PyMOL v.2.5.2分析VP7和VP4(VP8*)的中和表位。结果我国福建RVA毒株FJ21351116基因型为G3-P[8]-I2-R2-C2-M2-A2-N2-T2-E2-H2,系统进化分析显示,FJ21351116株的VP7、VP4、VP3、NSP2-NSP5基因与近几年日本检测到的马样DS-1样G3P[8]基因存在亲缘关系。而VP6、VP1、VP2、NSP1基因与大部分国家G2P[4]的相应基因亲缘关系近,特别是新加坡,表明该毒株是马样G3P[8]毒株与G2P[4]毒株共感染过程中通过基因重配形成的。FJ21351116的VP7/VP4基因与Rotarix和RotaTeq疫苗的进化分析表明,VP7和VP4(VP8*)中和抗原表位与疫苗氨基酸位点均存在多个突变。推测Rotarix和RotaTeq疫苗针对马样DS-1样G3P[8]RVA的保护效果不佳,且与Rotarix的中和抗原表位氨基酸差异高于RotaTeq。结论本研究发现一例中国罕见的DS-1样G3P[8]型RVA毒株,且疫苗株可能对其保护效果差,强调了持续监测RVA毒株及研发高效覆盖面全的RVA疫苗的重要性。展开更多
基金Supported by the German Federal Ministry of Education and Research(BMBF-Wachstumskern-PRAEMED.BIO),03WKDB2Csupported by the Janos Bolyai Research Scholarship of the Hungarian Academy of Sciences,BO/00232/17/5+1 种基金Research Grants of National Research Development and Innovation Office,K115818/2015/1New National Excellence Program of the Ministry of Human Capacities,ÚNKP-18-4 Bolyai Plus.
文摘BACKGROUND Defective neutrophil regulation in inflammatory bowel disease(IBD)is thought to play an important role in the onset or manifestation of IBD,as it could lead to damage of the intestinal mucosal barrier by the infiltration of neutrophils in the inflamed mucosa and the accumulation of pathogens.Like neutrophils in the context of innate immune responses,immunoglobulin A(IgA)as an acquired immune response partakes in the defense of the intestinal epithelium.Under normal conditions,IgA contributes to the elimination of microbes,but in connection with the loss of tolerance to chitinase 3-like 1(CHI3L1)in IBD,IgA could participate in CHI3L1-mediated improved adhesion and invasion of potentially pathogenic microorganisms.The tolerance brake to CHI3L1 and the occurrence of IgA autoantibodies to this particular target,the exact role and underlying mechanisms of CHI3L1 in the pathogenesis of IBD are still unclear.AIM To determine the predictive potential of Ig subtypes of a novel serological marker,anti-CHI3L1 autoantibodies(aCHI3L1)in determining the disease phenotype,therapeutic strategy and long-term disease course in a prospective referral cohort of adult IBD patients.METHODS Sera of 257 Crohn’s disease(CD)and 180 ulcerative colitis(UC)patients from a tertiary IBD referral center of Hungary(Division of Gastroenterology,Department of Internal Medicine,Faculty of Medicine,University of Debrecen)were assayed for IgG,IgA,and secretory IgA(sIgA)type aCHI3L1 by enzyme-linked immunosorbent assay using recombinant CHI3L1,along with 86 healthy controls(HCONT).RESULTS The IgA type was more prevalent in CD than in UC(29.2%vs 11.1%)or HCONT(2.83%;P<0.0001 for both).However,sIgA subtype aCHI3L1 positivity was higher in both CD and UC patients than in HCONT(39.3%and 32.8%vs 4.65%,respectively;P<0.0001).The presence of both IgA and sIgA aCHI3L1 antibodies was associated with colonic involvement(P<0.0001 and P=0.038,respectively)in patients with CD.Complicated disease behavior at sample procurement was associated with aCHI3L1 sIgA positivity(57.1%vs 36.0%,P=0.009).IgA type aCH3L1 was more prevalent in patients with frequent relapse during the disease course in the CD group(46.9%vs 25.7%,P=0.005).In a group of patients with concomitant presence of pure inflammatory luminal disease and colon involvement at the time of diagnosis,positivity for IgA or sIgA type aCH3L1 predicted faster progression towards a complicated disease course in time-dependent models.This association disappeared after merging subgroups of different disease locations.CONCLUSION CHI3L1 is a novel neutrophil autoantigenic target in IBD.The consideration of antibody classes along with location-based prediction may transform the future of serology in IBD.
基金Supported by Mr.Wade Thompson and family donation funds to the Divisions of Colon and Rectal surgery,Department of Surgery,Mount Sinai West Hospital,New York,NY 10019
文摘AIM: To assess blood chitinase 3-like 1(CHi3L1) levels for 2 mo after minimally invasive colorectal resection(MICR) for colorectal cancer(CRC). METHODS: CRC patients in an Institutional Review Board approved data/plasma bank who underwent elective MICR for whom preoperative(PreO p), early postoperative(PostO p), and 1 or more late PostO p samples [postoperative day(POD) 7-27] available were included. Plasma CHi3L1 levels(ng/m L) were determined in duplicate by enzyme linked immunosorbent assay. RESULTS: PreOp and PostOp plasma sample were available for 80 MICR cancer patients for the study. The median PreOp CHi3L1 level was 56.8 CI: 41.9-78.6 ng/mL(n = 80). Significantly elevated(P < 0.001) median plasma levels(ng/mL) over PreOp levels were detected on POD1(667.7 CI: 495.7, 771.7; n = 79), POD 3(132.6 CI: 95.5, 173.7; n = 76), POD7-13(96.4 CI: 67.7, 136.9; n = 62), POD14-20(101.4 CI: 80.7, 287.4; n = 22), and POD 21-27(98.1 CI: 66.8, 137.4; n = 20, P = 0.001). No significant difference in plasma levels were noted on POD27-41. CONCLUSION: Plasma CHi3L1 levels were significantly elevated for one month after MICR. Persistently elevated plasma CHi3L1 may support the growth of residual tumor and metastasis.
基金the Research Ethics Committee of The First Affiliated Hospital of Zhejiang University School of Medicine(No.2018-918).
文摘Background:Serum chitinase-3-like protein 1(CHI3L1)is a potential biomarker for fibrosis assessment.We aimed to evaluate serum CHI3L1 as a noninvasive diagnostic marker for chronic hepatitis B virusrelated fibrosis.Methods:Serum CHI3L1 levels were measured by ELISA in 134 chronic hepatitis B(CHB)patients.Significant fibrosis was defined as a liver stiffness>9.7 kPa.The performance of CHI3L1 was assessed and compared to that of other noninvasive tests by receiver operating characteristic(ROC)analysis.Results:Serum CHI3L1 levels were significantly higher in CHB patients with significant hepatic fibrosis(≥F2)than in those without significant hepatic fibrosis(<F2)(56.5 ng/mL vs.81.9 ng/mL,P<0.001).In CHB patients,the specificity and sensitivity of CHI3L1 for predicting significant fibrosis were 75.6%and 59.1%,respectively,with a cut-off of 76.0 ng/mL and an area under the ROC curve of 0.728(95%CI:0.637–0.820).Conclusions:Serum CHI3L1 levels could be an effective new serological biomarker for the diagnosis of liver fibrosis.Moreover,CHI3L1 is feasible in monitoring disease progression.
文摘AIM To identify whether chitinase 3-like 1(CHI3 L1) serves as a suitable biomarker for the prognosis of esophageal squamous cell carcinoma(ESCC) and to analyze this protein's cellular source.METHODS An ELISA was conducted to detect the concentration of CHI3 L1 in the serum of 150 ESCC patients diagnosed between January 2001 and February 2005. The prognostic relevance of CHI3 L1 was evaluated by a Kaplan-Meier and Cox regression analysis. The immunohistochemistry was reanalyzed,and fluorescent staining was utilized to explore the cellular origins of CHI3 L1. We stimulated monocyte-derived macrophages(MDMs) with either IL-6 or the supernatant of the ESCC cell line Eca-109 and later investigated the level of CHI3 L1 by q PCR and ELISA.RESULTS The level of serum CHI3 L1 was higher in older patients(≥ 60) than in patients under the age of 60(P = 0.001). The patients with higher levels of CHI3 L1 had a significantly shorter overall survival,whereas the traditional markers,carcinoembryonic antigen and squamous cell carcinoma antigen,were less effective(P > 0.05). A multivariate Cox analysis(P = 0.001) indicated that CHI3 L1 was an independent prognostic factor for ESCC patients. Peritumoral macrophages in ESCC exhibited high levels of CHI3 L1. Interleukin-6(IL-6) and the supernatant of Eca-109 containing IL-6 stimulated MDMs to secrete CHI3 L1. The serum concentration of CHI3 L1 in the ESCC patients showed a weak correlation with the laboratory inflammatory parameters neutrophil(NEU,P = 0.045),neutrophil/lymphocyte rate(NLR,P = 0.016),and C-reactive protein(CRP,P < 0.001).CONCLUSION Our study first established a connection between the pretreated CHI3 L1 and patients with ESCC,and the serum CHI3 L1 was primarily secreted by ESCC-surrounded macrophages.
基金Supported by the National Natural Science Foundation of China(No.81900829,No.82070963)the Xiamen Medical and Health Guiding Project Fund Project(No.3502Z20214ZD1214)+1 种基金the Guangdong Basic and Applied Basic Research Foundation(No.2019A1515011234)the Science and Technology Innovation Committee of Shenzhen(No.JCYJ20210324125614039)。
文摘AIM:To evaluate the potential of two trabecular meshwork(TM)-specific promoters,Chitinase 3-like 1(Ch3L1)and matrix gla protein(MGP),for improving specificity and safety in glaucoma gene therapy based on self-complementary AAV2(scAAV2)vector technologies.METHODS:An scAAV2 vector with C3 transferase(C3)as the reporter gene(scAAV2-C3)was selected.The scAAV2-C3 vectors were driven by Ch3L1(scAAV2-Ch3L1-C3),MGP(scAAV2-MGP-C3),enhanced MGP(scAAV2-eMGP-C3)and cytomegalovirus(scAAV2-CMV-C3),respectively.The cultured primary human TM cells were treated with each vector at different multiplicities of infections.Changes in cell morphology were observed by phase contrast microscopy.Actin stress fibers and Rho GTPases/Rho-associated protein kinase pathway-related molecules were assessed by immunofluorescence staining,real-time quantitative polymerase chain reaction and Western blot.Each vector was injected intracamerally into the one eye of each rat at low and high doses respectively.In vivo green fluorescence was visualized by a Micron III Retinal Imaging Microscope.Intraocular pressure(IOP)was monitored using a rebound tonometer.Ocular responses were evaluated by slit-lamp microscopy.Ocular histopathology analysis was examined by hematoxylin and eosin staining.RESULTS:In TM cell culture studies,the vectormediated C3 expression induced morphologic changes,disruption of actin cytoskeleton and reduction of fibronectin expression in TM cells by inhibiting the Rho GTPases/Rhoassociated protein kinase signaling pathway.At the same dose,these changes were significant in TM cells treated with scAAV2-CMV-C3 or scAAV2-Ch3L1-C3,but not in cells treated with scAAV2-eMGP-C3 or scAAV2-MGP-C3.At lowinjected dose,the IOP was significantly decreased in the scAAV2-Ch3L1-C3-injected eyes but not in scAAV2-MGPC3-injected and scAAV2-eMGP-C3-injected eyes.At highinjected dose,significant IOP reduction was observed in the scAAV2-eMGP-C3-injected eyes but not in scAAV2-MGP-C3-injected eyes.Similar to scAAV2-CMV-C3,scAAV2-Ch3L1-C3 vector showed efficient transduction both in the TM and corneal endothelium.In anterior segment tissues of scAAV2-eMGP-C3-injected eyes,no obvious morphological changes were found except for the TM.Inflammation was absent.CONCLUSION:In scAAV2-transduced TM cells,the promoter-driven efficiency of Ch3L1 is close to that of cytomegalovirus,but obviously higher than that of MGP.In the anterior chamber of rat eye,the transgene expression pattern of scAAV2 vector is presumably affected by MGP promoter,but not by Ch3L1 promoter.These findings would provide a useful reference for improvement of specificity and safety in glaucoma gene therapy using scAAV2 vector.
文摘目的探讨外周血类黏蛋白1样蛋白3(ORMDL3)水平与过敏性哮喘(AA)患儿Th1/Th2平衡的关系。方法选取64例AA患儿为AA组,另选取48例同时期体检健康儿童为对照组;检测两组血清Ig E、嗜酸性粒细胞比率(EOSR),外周血ORMDL3 m RNA、Th1/Th2水平,以及呼出气一氧化氮(Fe NO)、达峰容积比(VPEF/VE)、达峰时间比(TPTEF/TE)。Pearson相关性分析AA组外周血ORMDL3 m RNA与Th1/Th2水平及两者与Ig E、EOSR、Fe NO、VPEF/VE、TPTEF/TE的相关性,多因素Logistics回归分析AA危险因素。结果AA组血清Ig E、EOSR、Fe NO和外周血ORMDL3 m RNA、Th2水平高于对照组,VPEF/VE、TPTEF/TE、Th2水平和Th1/Th2低于对照组(P均<0.05);AA患儿外周血ORMDL3 m RNA与Th1/Th2水平呈负相关(r=-7.103,P<0.05)。外周血ORMDL3 m RNA水平与Ig E、EOSR、Fe NO呈正相关,与VPEF/VE、TPTEF/TE呈负相关(P均<0.05),Th1/Th2水平与Ig E、EOSR、Fe NO呈负相关,与VPEF/VE、TPTEF/TE呈正相关(P均<0.05)。多因素Logistics回归分析显示,Ig E(OR=1.103,95%CI=1.019~1.193)和ORMDL3 m RNA(OR=13.697,95%CI=2.203~15.161)为AA发生独立危险因素,Th1/Th2(OR=0.040,95%CI=0.004~0.396)为保护因素(P均<0.05)。结论AA患儿外周血ORMDL3 m RNA水平明显升高,Th1/Th2处失衡状态,二者可作为评估AA病情的指标。
基金the Science and Technology Innovation Talents of Shenyang,China(Grant No.RC190489).
文摘Sheath blight disease (ShB) has a severe impact on the production of rice. ABI3/VP1-like 1(RAVL1) negatively regulated the rice defense mechanism against ShB, however, this regulatorymechanism is not clearly understood. In this study, we identified that indeterminate domain 3 (IDD3) waspositively regulated by RAVL1. Further, chromatin immunoprecipitation (ChIP) assay, yeast one-hybridassay and transient expression assay indicated a direct binding between RAVL1 and the IDD3 promoterregion. IDD3 was ubiquitously expressed in different tissues and at different stages, and its expressionwas significantly enhanced by Rhizoctonia solani infection. IDD3 exhibited transcription activation activityin yeast and IDD3-GFP was found to be localized in the nucleus. IDD3 mutants exhibited no significantdifferences in response to ShB, while IDD3 overexpressors were more susceptible to ShB compared withwild type (WT) plants. Furthermore, IDD3 repressors were less susceptible to R. solani than WT plants.Interestingly, the expression of brassinosteroid-related genes (D2, D11 and BRI1) was lower in IDD3repressors and higher in IDD3 overexpressors compared with WT. However, the ChIP assay revealedthat IDD3 did not directly bind to the D2 and D11 promoters. Overexpression of IDD3 in BRI1 mutantd61-1 inhibited the activity of IDD3, reducing its susceptibility to ShB compared with IDD3 overexpressorand WT plants, indicating that IDD3 negatively regulated the rice defense mechanism against ShB by activatingthe BR signaling pathway. Thus, our analyses provided information to enhance the understanding of therice defense mechanism against ShB.
基金supported by grants from the Thirteen-Fifth Mega-Scientific Project on“Prevention and Treatment of AIDS,Viral Hepatitis and Other Infectious Diseases”(No.2017ZX10201301-007-002)the National Natural Science Foundation of China(No.82072233).
文摘Objective Current commercially available immunological tests cannot be used for discriminating active tuberculosis(TB)from latent TB infection.To evaluate the value of biomarker candidates in the diagnosis of active TB,this study aimed to identify differentially expressed genes in peripheral blood mononuclear cells(PBMCs)between patients with active TB and individuals with latent TB infection by transcriptome sequencing.Methods The differentially expressed genes in unstimulated PBMCs and in Mycobacterium tuberculosis(Mtb)antigen-stimulated PBMCs from patients with active TB and individuals with latent TB infection were identified by transcriptome sequencing.Selected candidate genes were evaluated in cohorts consisting of 110 patients with TB,30 individuals with latent TB infections,and 50 healthy controls by quantitative real-time RT-PCR.Receiver operating characteristic(ROC)curve analysis was performed to calculate the diagnostic value of the biomarker candidates.Results Among the differentially expressed genes in PBMCs without Mtb antigen stimulation,interferon-induced protein with tetratricopeptide repeats 3(IFIT3)had the highest area under curve(AUC)value(0.918,95%CI:0.852-0.984,P<0.0001)in discriminating patients with active TB from individuals with latent TB infection,with a sensitivity of 91.86%and a specificity of 84.00%.In Mtb antigen-stimulated PBMCs,orosomucoid 1(ORM1)had a high AUC value(0.833,95%CI:0.752-0.915,P<0.0001),with a sensitivity of 81.94%and a specificity of 70.00%.Conclusion IFIT3 and ORM1 might be potential biomarkers for discriminating active TB from latent TB infection.
文摘目的对2021年采集于我国福建的A组轮状病毒(group A rotavirus,RVA)G3P[8]毒株FJ21351116进行全基因组分子特征分析。方法使用高灵敏度A组轮状病毒全基因组测序方法对FJ21351116进行全基因组测序。用MEGA11.0、Geneious9.0.2和DNASTAR软件通过核酸序列分析评估病毒的基因组特征。使用BioEdit v.7.0.9.0和PyMOL v.2.5.2分析VP7和VP4(VP8*)的中和表位。结果我国福建RVA毒株FJ21351116基因型为G3-P[8]-I2-R2-C2-M2-A2-N2-T2-E2-H2,系统进化分析显示,FJ21351116株的VP7、VP4、VP3、NSP2-NSP5基因与近几年日本检测到的马样DS-1样G3P[8]基因存在亲缘关系。而VP6、VP1、VP2、NSP1基因与大部分国家G2P[4]的相应基因亲缘关系近,特别是新加坡,表明该毒株是马样G3P[8]毒株与G2P[4]毒株共感染过程中通过基因重配形成的。FJ21351116的VP7/VP4基因与Rotarix和RotaTeq疫苗的进化分析表明,VP7和VP4(VP8*)中和抗原表位与疫苗氨基酸位点均存在多个突变。推测Rotarix和RotaTeq疫苗针对马样DS-1样G3P[8]RVA的保护效果不佳,且与Rotarix的中和抗原表位氨基酸差异高于RotaTeq。结论本研究发现一例中国罕见的DS-1样G3P[8]型RVA毒株,且疫苗株可能对其保护效果差,强调了持续监测RVA毒株及研发高效覆盖面全的RVA疫苗的重要性。