Prognostic and Pathophysiologic Significance of IL-8 (CXCL8) in Biliary Atresia

Interleukin (IL)-8 (CXCL8), a chemokine involved in neutrophil recruitment, has been implicated in ductular reaction and liver fibrogenesis. We studied liver and serum IL-8 expression in a large biliary atresia (BA) cohort and explored its prognostic and pathophysiological potential. IL-8 expression was assessed in liver utilizing quantitative polymerase chain reaction (qPCR), immunohistochemistry and in situ hybridization and in serum using an enzyme-linked immunosorbent assay, among 115 BA patients, 10 disease controls and 68 normal controls. Results were correlated to portoenterostomy (PE) outcomes, biochemical and histological liver injury, transcriptional markers of fibrosis and cholangiocytes, and expression of other related cytokines. IL-8 was markedly overexpressed in liver and serum of BA patients at PE (n = 88) and in serum samples obtained during postoperative follow-up (n = 40). IL-8 expression in the liver was predominantly in cholangiocytes within areas of ductular reaction. Liver IL-8 mRNA expression correlated positively with its serum concentration, bile ductular proliferation, Metavir fibrosis stage, and transcriptional markers of activated myofibroblasts (ACTA2) and cholangiocytes (KRT19). Taken together, IL-8 may mediate liver injury in BA by promoting ductular reaction and associated liver fibrogenesis. Prognostic value of serum IL-8 to predict native liver survival was limited and confined to the postoperative period after PE.

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miR-128-3p inhibits apoptosis and inflammation in LPS-induced sepsis by targeting TGFBR2

Open Medicine ◽

10.1515/med-2021-0222 ◽

2021 ◽

Vol 16 (1) ◽

pp. 274-283

Author(s):

Peng Yang ◽

Jianhua Han ◽

Shigeng Li ◽

Shaoning Luo ◽

Xusheng Tu ◽

...

Keyword(s):

Transforming Growth Factor Beta ◽

Transforming Growth Factor ◽

Quantitative Polymerase Chain Reaction ◽

Luciferase Reporter ◽

Enzyme Linked Immunosorbent Assay ◽

Serum Samples ◽

Aberrant Expression ◽

Protein Levels ◽

Apoptosis And Inflammation ◽

Hk2 Cells

Abstract Background Sepsis is a systemic inflammatory response that can lead to the dysfunction of many organs. The aberrant expression of miRNAs is associated with the pathogenesis of sepsis. However, the biological functions of miR-128-3p in sepsis remain largely unknown, and its mechanism should be further investigated. This study aimed to determine the regulatory network of miR-128-3p and TGFBR2 in lipopolysaccharide (LPS)-induced sepsis. Methods The expression levels of miR-128-3p and transforming growth factor beta receptors II (TGFBR2) were detected by quantitative polymerase chain reaction (qPCR). The protein levels of TGFBR2, Bcl-2, Bax, cleaved caspase 3, Smad2, and Smad3 were measured by western blot. Cell apoptosis was analyzed by flow cytometry. Cytokine production was detected by enzyme-linked immunosorbent assay (ELISA). The binding sites of miR-128-3p and TGFBR2 were predicted by Targetscan online software and confirmed by dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay. Results The level of miR-128-3p was decreased, and TGFBR2 expression was increased in serum samples of sepsis patients and LPS-induced HK2 cells. Overexpression of miR-128-3p or knockdown of TGFBR2 ameliorated LPS-induced inflammation and apoptosis. Moreover, TGFBR2 was a direct target of miR-128-3p, and its overexpression reversed the inhibitory effects of miR-128-3p overexpression on inflammation and apoptosis in LPS-induced HK2 cells. Besides, overexpression of miR-128-3p downregulated TGFBR2 to suppress the activation of the Smad signaling pathway. Conclusion miR-128-3p could inhibit apoptosis and inflammation by targeting TGFBR2 in LPS-induced HK2 cells, which might provide therapeutic strategy for the treatment of sepsis.

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Dynamic Analysis of Serum MMP-7 and Its Relationship with Disease Progression in Biliary Atresia: A Multicenter Prospective Study

10.21203/rs.3.rs-866431/v1 ◽

2021 ◽

Author(s):

Shuiqing Chi ◽

Peipei Xu ◽

Pu Yu ◽

Guoqing Cao ◽

Haibin Wang ◽

...

Keyword(s):

Prospective Study ◽

Biliary Atresia ◽

Disease Progression ◽

Genetic Mutation ◽

Mediation Effect ◽

Enzyme Linked Immunosorbent Assay ◽

High Concentration ◽

Clinical Value ◽

Serum Samples ◽

Matrix Metalloproteinase 7

Abstract PurposeWe aimed to assess the dynamic changing trend of serum matrix metalloproteinase-7 (MMP-7) in BA patients from diagnosis to LTx to further elucidate its clinical value in diagnosis and prognoses and its relationship with disease progression.MethodsIn this multicentre prospective study, a total of 440 cholestasis patients (direct bilirubin level of > 17 μmol/L) were enrolled. Serum and stool MMP-7 levels were measured using an enzyme-linked immunosorbent assay at different time points and analysed. The polymorphism of MMP-7 was assessed to explore the possible reasons for the low value in BA patients.Results:In neonate biliary atresia patients, using a cut-off value of > 26.73 ng/ml, serum MMP-7 had a AUC of 0.954. A genetic mutation (G137D) and rapid degradation of MMP-7 in serum samples could cause low MMP-7 levels in serum of BA patients. Four dynamic patterns of serum MMP-7 post-KPE were associated with prognosis. A high concentration of MMP-7 in the stool is linked to a decreased serum MMP-7 concentration. MMP-7 showed a mediation effect on the association between inflammation and liver fibrosis in BA patients. Serum MMP-7 was the only significant predictor at six weeks post-KPE and the most accurate predictor at three months post-KPE of survival with the native liver (SNL) in two years.Conclusion:As one of the critical factors associated with BA occurrence and progression, serum MMP-7 can be used for early diagnosis of BA and post-KPE MMP-7 level is the earliest prognostic biomarker so far.

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A Naturally Transmitted Epitheliotropic Polyomavirus Pathogenic in Immunodeficient Rats: Characterization, Transmission, and Preliminary Epidemiologic Studies

Toxicologic Pathology ◽

10.1177/0192623317723541 ◽

2017 ◽

Vol 45 (5) ◽

pp. 593-603 ◽

Cited By ~ 8

Author(s):

Cynthia Besch-Williford ◽

Patricia Pesavento ◽

Shari Hamilton ◽

Beth Bauer ◽

Beatrix Kapusinszky ◽

...

Keyword(s):

Quantitative Polymerase Chain Reaction ◽

Prostate Gland ◽

Interleukin 2 ◽

Epidemiologic Studies ◽

Metagenomic Sequencing ◽

Serum Samples ◽

Diagnostic Assays ◽

Wu Polyomavirus ◽

Washington University

We report the identification, pathogenesis, and transmission of a novel polyomavirus in severe combined immunodeficient F344 rats with null Prkdc and interleukin 2 receptor gamma genes. Infected rats experienced weight loss, decreased fecundity, and mortality. Large basophilic intranuclear inclusions were observed in epithelium of the respiratory tract, salivary and lacrimal glands, uterus, and prostate gland. Unbiased viral metagenomic sequencing of lesioned tissues identified a novel polyomavirus, provisionally named Rattus norvegicus polyomavirus 2 (RatPyV2), which clustered with Washington University (WU) polyomavirus in the Wuki clade of the Betapolyomavirus genus. In situ hybridization analyses and quantitative polymerase chain reaction (PCR) results demonstrated viral nucleic acids in epithelium of respiratory, glandular, and reproductive tissues. Polyomaviral disease was reproduced in Foxn1rnu nude rats cohoused with infected rats or experimentally inoculated with virus. After development of RatPyV2-specific diagnostic assays, a survey of immune-competent rats from North American research institutions revealed detection of RatPyV2 in 7 of 1,000 fecal samples by PCR and anti-RatPyV2 antibodies in 480 of 1,500 serum samples. These findings suggest widespread infection in laboratory rat populations, which may have profound implications for established models of respiratory injury. Additionally, RatPyV2 infection studies may provide an important system to investigate the pathogenesis of WU polyomavirus diseases of man.

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Prevalence of Antibodies to Onchocerca volvulus in Residents of Oaxaca, Mexico, Treated for 10 Years with Ivermectin

Clinical and Diagnostic Laboratory Immunology ◽

10.1128/cdli.12.1.40-43.2005 ◽

2005 ◽

Vol 12 (1) ◽

pp. 40-43 ◽

Cited By ~ 1

Author(s):

Alberto Gómez-Priego ◽

Raymundo Mendoza ◽

Jorge-Luis de-la-Rosa

Keyword(s):

Crude Extract ◽

Onchocerca Volvulus ◽

Enzyme Linked Immunosorbent Assay ◽

Ivermectin Treatment ◽

Antibody Prevalence ◽

Serum Samples ◽

Igg Antibody ◽

Prevalence Studies ◽

Serological Studies

ABSTRACT Studies to determine the prevalence of antibodies to Onchocerca volvulus, prior to and after actions carried out to interrupt transmission, are scarce in Mexico. Here we report the prevalence of immunoglobulin G (IgG) and IgG4 antibodies in an enzyme-linked immunosorbent assay (ELISA) against a crude extract of O. volvulus adult worm in serum samples from persons under noninterrupted biannual treatment with ivermectin in areas of onchocercosis endemicity in Mexico. To perform the prevalence studies, the ELISA procedures were first evaluated. Serological studies were performed with serum samples from skin microfilaria carriers from Guatemala and from people microfilariodermic negative living in the same area as the Guatemalan patients. Sensitivity values for IgG or IgG4 detection were 71 and 86%, while specificities were 92 and 100%, respectively. No anti-O. volvulus antibodies were found in samples from nonendemic controls from Mexico, but 3 of 71 samples from residents in the onchocercosis area of Oaxaca, Mexico, and who have been under ivermectin treatment during the last 10 years were only positive to IgG. Notwithstanding that the IgG4 isotype was not detected and a low (4.2%) anti-O. volvulus IgG antibody prevalence was found, a seroepidemiological follow-up must be performed in order to confirm interruption of onchocercosis transmission in the area of Oaxaca, Mexico, in which onchocercosis is endemic.

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Validation of a Blocking Enzyme-Linked Immunosorbent Assay for Detection of Antibodies against Porcine Reproductive and Respiratory Syndrome Virus

Clinical and Diagnostic Laboratory Immunology ◽

10.1128/cdli.11.3.503-514.2004 ◽

2004 ◽

Vol 11 (3) ◽

pp. 503-514 ◽

Cited By ~ 22

Author(s):

Neal H. Ferrin ◽

Ying Fang ◽

Craig R. Johnson ◽

Michael P. Murtaugh ◽

Dale D. Polson ◽

...

Keyword(s):

Immunosorbent Assay ◽

Fluorescent Antibody ◽

Internal Quality ◽

Respiratory Syndrome Virus ◽

Enzyme Linked Immunosorbent Assay ◽

Characteristic Analysis ◽

Serum Samples ◽

Negative Results ◽

Control Serum

ABSTRACT Porcine reproductive and respiratory syndrome (PRRS) continues to be one of the most significant diseases of swine. IDEXX HerdChek PRRS, a commercially available enzyme-linked immunosorbent assay (ELISA), has become the industry standard for the detection of antibodies against PRRS virus (PRRSV). The need to accurately determine the PRRSV serostatus of herds and individual animals has prompted the development of several follow-up assay methods. A highly specific and repeatable blocking ELISA (bELISA) was developed on the basis of the use of an expressed PRRSV nucleocapsid (N) protein as the antigen and a biotinylated monoclonal antibody. Validation of the bELISA used sera from 316 animals experimentally and naturally infected with North American PRRSV and sera from 370 uninfected animals. Receiver operating characteristic analysis of the data calculated a diagnostic sensitivity of 97.8% and a diagnostic specificity of 100%. The between-run coefficient of variation of an internal quality control serum was 4.24%. The bELISA was able to detect seroconversion as well as the IDEXX ELISA and the indirect fluorescent antibody (IFA) assay; kappa values were 0.94 and 0.96, respectively. A collection of 133 serum samples with unexpected positive IDEXX ELISA results was obtained from 4,038 diagnostic samples submitted from farms from which PRRS-negative results were expected. The bELISA identified 97% of the samples as PRRS seronegative, while the IFA identified 100% as seronegative. The anticipated use of the bELISA is as a follow-up test to the IDEXX ELISA for determining the PRRSV serostatus of individual animals with unexpected positive test results from swine herds from which negative results are expected.

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A Colorimetric Enzyme-Linked Immunosorbent Assay with CuO Nanoparticles as Signal Labels Based on the Growth of Gold Nanoparticles In Situ

Nanomaterials ◽

10.3390/nano9010004 ◽

2018 ◽

Vol 9 (1) ◽

pp. 4 ◽

Cited By ~ 3

Author(s):

Dehua Deng ◽

Yuanqiang Hao ◽

Jiajia Xue ◽

Xiuhua Liu ◽

Xinyue Xu ◽

...

Keyword(s):

Gold Nanoparticles ◽

Fluorescence Probe ◽

Colorimetric Assay ◽

Specific Antigen ◽

Cuo Nanoparticles ◽

Enzyme Linked Immunosorbent Assay ◽

Serum Samples ◽

Cuo Nps ◽

Colorimetric Immunoassay

A colorimetric immunoassay has been reported for prostate-specific antigen (PSA) detection with CuO nanoparticles (CuO NPs) as signal labels. The method is based on Cu2+-catalyzed oxidation of ascorbic acid (AA) by O2 to depress the formation of colored gold nanoparticles (AuNPs). Specifically, HAuCl4 can be reduced by AA to produce AuNPs in situ. In the presence of target, CuO NPs-labeled antibodies were captured via the sandwich-type immunoreaction. After dissolving CuO nanoparticles with acid, the released Cu2+ catalyzed the oxidation of AA by O2, thus depressing the generation of AuNPs. To demonstrate the accuracy of the colorimetric assay, the released Cu2+ was further determined by a fluorescence probe. The colorimetric immunoassay shows a linear relationship for PSA detection in the range of 0.1~10 ng/mL. The detection limit of 0.05 ng/mL is comparable to that obtained by other CuO NPs-based methods. The high throughput, simplicity, and sensitivity of the proposed colorimetric immunoassay exhibited good applicability for assays of serum samples.

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Measurement of Connective Tissue Parameters in Serum for Diagnosis and Follow-Up of Liver Fibrosis

Annals of Clinical Biochemistry International Journal of Laboratory Medicine ◽

10.1177/000456328702400308 ◽

1987 ◽

Vol 24 (3) ◽

pp. 283-292 ◽

Cited By ~ 14

Author(s):

A M Gressner

Keyword(s):

Molecular Weight ◽

Liver Fibrosis ◽

Liver Injury ◽

Serum Level ◽

Connective Tissue ◽

Venous Pressure ◽

Membrane Glycoprotein ◽

Chronic Alcoholic ◽

Liver Fibrogenesis

Hepatic fibrogenesis, i.e. activated synthesis and excessive intercellular deposition of connective tissue molecules (collagens, adhesive glycoproteins, proteoglycans) occurs in chronic alcoholic and viral liver injury and, less frequently, in some other conditions. The process may be monitored biochemically by the radioimmunoassay of some connective tissue molecules or their fragments and by the measurement of the activity of certain enzymes in serum. Currently, the radioimmunoassay of the aminoterminal propeptide of type III procollagen in serum reflects best the activity of liver fibrogenesis. The serum level of laminin, a high molecular weight basement membrane glycoprotein, was found to be correlated with an elevated portal venous pressure.

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Immunochromatographic Test with Recombinant Em18 Antigen for the Follow-Up Study of Alveolar Echinococcosis

Clinical and Vaccine Immunology ◽

10.1128/cvi.05156-11 ◽

2011 ◽

Vol 18 (8) ◽

pp. 1302-1305 ◽

Cited By ~ 15

Author(s):

Yasuhito Sako ◽

Dennis Tappe ◽

Kenta Fukuda ◽

Yukuharu Kobayashi ◽

Sonoyo Itoh ◽

...

Keyword(s):

Alveolar Echinococcosis ◽

Enzyme Linked Immunosorbent Assay ◽

Immunochromatographic Test ◽

Serum Samples ◽

Content Type ◽

Follow Up Study ◽

The Status ◽

Antibody Levels ◽

Kinetics Of

ABSTRACTThe performance of a rapid and simple immunochromatographic test (ICT) with recombinant Em18 (rEm18) antigen for serological follow-up ofEchinococcus multilocularisinfection was evaluated by comparison with that of an enzyme-linked immunosorbent assay (ELISA) with rEm18, using serum samples from patients who underwent surgery and/or received antiparasitic chemotherapy. The degree of Em18-band intensity on the ICT correlated highly with the absorbance value obtained by the ELISA. The kinetics of antibody levels obtained by the ICT paralleled those of the ELISA. These data suggest that the ICT has high potential as an easy-to-handle, fast, and reliable follow-up tool to monitor the status of alveolar echinococcosis in different stages.

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LncRNA HCP5 knockdown inhibits high glucose-induced excessive proliferation, fibrosis and inflammation of human glomerular mesangial cells by regulating the miR-93-5p/HMGA2 axis

BMC Endocrine Disorders ◽

10.1186/s12902-021-00781-y ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Xuan Wang ◽

Yan Liu ◽

Jian Rong ◽

Kai Wang

Keyword(s):

High Glucose ◽

Mesangial Cells ◽

Quantitative Polymerase Chain Reaction ◽

Cell Counting ◽

Luciferase Reporter ◽

Enzyme Linked Immunosorbent Assay ◽

Inflammatory Factor ◽

Glomerular Mesangial Cells ◽

Serum Samples ◽

Human Glomerular Mesangial Cells

Abstract Background Long non-coding RNAs (lncRNAs) are widely reported to be involved in the development of human diseases. HLA complex P5 (HCP5) deregulation is associated with various diseases. However, the function of HCP5 in diabetic nephropathy (DN) is unclear. Methods Human glomerular mesangial cells (HGMCs) were treated with high glucose (HG) to establish DN cell models. The expression of HCP5, miR-93-5p and high mobility group AT-hook 2 (HMGA2) mRNA was detected using quantitative polymerase chain reaction (QPCR). Cell proliferation and cell apoptosis were assessed using cell counting kit-8 (CCK-8) assay and flow cytometry assay, respectively. The expression of apoptosis- and fibrosis-related proteins and HMGA2 protein was quantified by western blot. The release of pro-inflammatory factor was checked using enzyme-linked immunosorbent assay (ELISA). The predicted relationship between miR-93-5p and HCP5 or HMGA2 was verified using dual-luciferase reporter assay, pull-down assay or RNA immunoprecipitation (RIP) assay. Results The expression of HCP5 and HMGA2 was enhanced, while the expression of miR-93-5p was declined in DN serum samples and HG-treated HGMCs. HCP5 knockdown or miR-93-5p restoration ameliorated HG-induced HGMC proliferation, fibrosis and inflammation. MiR-93-5p was a target of HCP5, and miR-93-5p inhibition reversed the effects caused by HCP5 knockdown. Moreover, HMGA2 was a target of miR-93-5p, and HMGA2 overexpression abolished the effects of miR-93-5p restoration. HCP5 knockdown inhibited the AKT/mTOR signaling pathway. Conclusion HCP5 was implicated in DN progression by modulating the miR-93-5p/HMGA2 axis, which provided new insights into the understanding of DN pathogenesis.

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