Screening of serological markers for differential diagnosis ischemic colitis and ulcerative colitis by proteomic techniques / 中华消化杂志

Objective To screen and identify serum protein biomarkers for the differential diagnosis between ischemic colitis (IC) and ulcerative colitis (UC) by tandem mass tag (TMT) combined with liquid chromatography/tandem mass spectrometry (LC-MS/MS).Methods From January 2018 to January 2019,at the First Affiliated Hospital of School of Medicine of Zhejiang University,patients with UC or IC,and health controls,each l0 cases,were enrolled into UC group,IC group and normal control (NC) group,respectively.Fasting serum samples of all the subjects were collected.After removal of high-abundance protein,followed by proteolysis,peptide labeling and fractionating,the samples were then processed by mass spectrometry.The protein with TMT data of three groups was obtained and protein with TMT value 0 were removed.Heat map of protein was constructed.The differential protein was defined as the protein fold change over 1.5 or less than 0.67.The Reactome database was used to cluster the pathways of differential proteins among groups.Statistical methods included t test,hypergeometry test and corrected by BH multiple test.Results A total of 357 serum proteins were identified by proteomic profiling.There were 27 differential proteins between the IC group and the NC group,including six up-regulated proteins and 21 down-regulated proteins.There were 228 differential proteins between the UC group and the NC group,including 75 up-regulated proteins and 153 down-regulated proteins.There were 49 differential proteins between UC group and IC group,including 22 up-regulated proteins and 27 down-regulated proteins.In the comparison of differential proteins between the NC group,IC group and UC group,only the expression of fibrin 3 was statistically significant (the fold change between UC and NC,between UC and IC,between IC and NC were 0.24,0.46 and 0.53,respectively;t =-5.089,-7.298 and -3.919,all P ＜ 0.01).The results of pathway cluster analysis showed that in the comparison of differential proteins between IC group and NC group,only the platelet degranulation pathway was enriched,and 10 proteins were involved in this pathway (P ＜ 0.01).In the comparison of differential proteins between UC group and NC group,there were 58 pathways enriched,of which 38 proteins were involved in the platelet degranulation pathway,16 proteins were involved in the initial complement trigger pathway,13 proteins were involved in the complement cascade pathway,and 11 proteins were involved in antibody-mediated complement activation pathway (all P ＜ 0.01).In the comparison of differential proteins between UC group and IC group,three different pathways were obtained.Among them,nine proteins were involved in the platelet degranulation pathway,seven proteins were involved in the initial complement trigger pathway,and five proteins were involved in the complement cascade pathway (all P ＜ 0.01).Conclusions The difference in serum proteome between IC patients and UC patients was significant,and the differential proteins are mainly involved in platelet activation and complement activation.The candidate proteins identified in this study may be used as biomarkers for the differential diagnosis of UC and IC in the future.

Screening of serological markers for differential diagnosis ischemic colitis and ulcerative colitis by proteomic techniques / 中华消化杂志

Objective@#To screen and identify serum protein biomarkers for the differential diagnosis between ischemic colitis (IC) and ulcerative colitis (UC) by tandem mass tag (TMT) combined with liquid chromatography/tandem mass spectrometry (LC-MS/MS).@*Methods@#From January 2018 to January 2019, at the First Affiliated Hospital of School of Medicine of Zhejiang University, patients with UC or IC, and health controls, each 10 cases, were enrolled into UC group, IC group and normal control (NC) group, respectively. Fasting serum samples of all the subjects were collected. After removal of high-abundance protein, followed by proteolysis, peptide labeling and fractionating, the samples were then processed by mass spectrometry. The protein with TMT data of three groups was obtained and protein with TMT value 0 were removed. Heat map of protein was constructed. The differential protein was defined as the protein fold change over 1.5 or less than 0.67. The Reactome database was used to cluster the pathways of differential proteins among groups. Statistical methods included t test, hypergeometry test and corrected by BH multiple test.@*Results@#A total of 357 serum proteins were identified by proteomic profiling. There were 27 differential proteins between the IC group and the NC group, including six up-regulated proteins and 21 down-regulated proteins. There were 228 differential proteins between the UC group and the NC group, including 75 up-regulated proteins and 153 down-regulated proteins. There were 49 differential proteins between UC group and IC group, including 22 up-regulated proteins and 27 down-regulated proteins. In the comparison of differential proteins between the NC group, IC group and UC group, only the expression of fibrin 3 was statistically significant (the fold change between UC and NC, between UC and IC, between IC and NC were 0.24, 0.46 and 0.53, respectively; t=-5.089, -7.298 and -3.919, all P<0.01). The results of pathway cluster analysis showed that in the comparison of differential proteins between IC group and NC group, only the platelet degranulation pathway was enriched, and 10 proteins were involved in this pathway (P<0.01). In the comparison of differential proteins between UC group and NC group, there were 58 pathways enriched, of which 38 proteins were involved in the platelet degranulation pathway, 16 proteins were involved in the initial complement trigger pathway, 13 proteins were involved in the complement cascade pathway, and 11 proteins were involved in antibody-mediated complement activation pathway (all P<0.01). In the comparison of differential proteins between UC group and IC group, three different pathways were obtained. Among them, nine proteins were involved in the platelet degranulation pathway, seven proteins were involved in the initial complement trigger pathway, and five proteins were involved in the complement cascade pathway (all P<0.01).@*Conclusions@#The difference in serum proteome between IC patients and UC patients was significant, and the differential proteins are mainly involved in platelet activation and complement activation. The candidate proteins identified in this study may be used as biomarkers for the differential diagnosis of UC and IC in the future.

Clinical significance of detecting anti-Saccharomyces cerevisiae antibody in Crohn′s disease / 中华消化杂志

Objective To evaluate the clinical significance of detecting serum anti-Saccharomyces cerevisiae antibody (ASCA)IgG and IgA in the diagnosis of Crohn′s disease(CD).Methods A total of 51 patients with CD were enrolled as CD group and 22 healthy volunteers as healthy control group.The serum samples of both groups were collected.ASCA-IgG and ASCA-IgA were determined with enzyme linked immunosorbent assay (ELISA).According to Montreal standard,patients with CD were divided into subgroup according to the age of onset (A),lesion (L),clinical behavior (B).The sensitivity, specificity and positive predictive value of both groups were calculated.Chi-square test was performed for count data analysis.Results The sensitivities of ASCA-IgG and ASCA-IgA in CD group were 45 .1 % and 35 .3%,respectively,while in the healthy control group which were 0 and 9.1 %,respectively.There were significant differences between two groups (χ2 =14.49 and 5 .31 ,both P 0.05 ).The sensitivities of ASCA-IgG in CD patients with complications and without complications were 56.3% and 26.3%,respectively,and there was significant difference (χ2 =4.31 ,P <0.05).Conclusions Serum ASCA-IgG is not suitable for population screening,however it has certain value for the differential diagnosis of CD.The clinical value of detecting ASCA-IgG is higher than that of detecting ASCA-IgA.

Fundamento biológico y aplicación clínica de los marcadores tumorales séricos / Biological basis and clinical application of serum tumor markers

El cáncer es el resultado de la acumulación de alteraciones en moléculas con importante función en procesos celulares como proliferación, apoptosis, muerte celular y reparación génica. Las moléculas, sustancias o procesos alterados pueden constituirse en marcadores o biomarcadores tumorales de gran utilidad clínica en el seguimiento de pacientes oncológicos ya que han demostrado ser idóneos para la valoración del tratamiento y su eficiencia. La determinación de biomarcadores tumorales no ha sido muy exitosa debido a la baja sensibilidad y especificidad de las técnicas usadas y al requerimiento de muestras biológicas en volúmenes grandes o de métodos invasivos para su recolección. Los marcadores tumorales séricos surgen, entonces, como una herramienta útil en la obtención de información sobre el estado de la enfermedad y constituye un reto científico mejorar su aplicabilidad en el diagnóstico temprano, pronóstico, seguimiento de la enfermedad y evaluación de la eficacia terapéutica.

Cancer is the result of the accumulation of changes in molecules with important functions in processes such as cell proliferation, apoptosis, cell death and gene repair. Molecules, substances or altered pathways constitute tumor markers or biomarkers useful in clinical monitoring of cancer patients, because they have demonstrated to be suitable for the valuation of the patient's treatment and it efficiency. Determination of tumor markers has not been very successful due to the low sensitivity and specificity of the techniques used and the requirement of large volumes of biological samples or the use of invasive methods for collecting them. The serum tumor markers arise, as a useful tool to obtain information about the disease progress and constitute as a scientific challenge to improve its applicability in early diagnosis, prognosis, monitoring of the disease and evaluation of therapeutic efficacy.

Proteomic identificafion of secreted protein DDH in non-small cell lung cancer / 肿瘤

Objective: To identify secreted proteins of non-small cell lung cancer (NSCLC) and provide candidate molecule for screening serological biomarkers of lung cancer. Methods: NSCLC cell line A 549 cells were cultured. The proteins in the conditioned medium were collected and identified by proteomic analysis and spectrometry. Identified protein was confirmed by Western blotting in the conditioned medium. Fifteen NSCLC tissues and paired distant lung tissues were obtained simultaneously during surgery. The expression difference between cancer tissues and non-malignant lung tissues were analyzed by RT-PCR, Western blotting, and immunochemistry. One hundred and four cases of serum specimens were collected from NSCLC patients (n = 64), non-malignant lung tumor patients (n = 20) and healthy controls (n = 20). Serum level of dehydrodiol dehydrogenase (DDH) was analyzed by ELISA. Results: DDH was identified by proteomic approach and confirmed by Western blotting in the conditioned medium of A 549 cells. RT-PCR, Western blot, and immunochemistry analysis showed that over-expression of DDH mRNA and protein were detected in NSCLC cancer tissues compared with distant non-malignant lung tissues (P < 0.05). Abundant DDH was observed in cytoplasm and membrane of lung cancer cells. The serum level of DDH was significantly higher in NSCLC patients than that in non-malignant lung tumor patients and healthy controls (P < 0. 05). Conclusion: DDH is over-expressed in lung cancer tissues and the serum level of DDH is elevated in NSCLC patients. The secreted DDH protein is a serological biomarker of NSCLC.