1.Establishment and preliminary validation of a diagnostic prediction model for esophageal squamous cell carcinoma based on MALDI-TOF MS
Ting ZHANG ; Nana WANG ; Lu YANG ; Yingjian CHEN ; Chengjin HU
Chinese Journal of Clinical Laboratory Science 2019;37(2):81-86
Objective:
To establish a diagnostic prediction model for esophageal squamous cell carcinoma (ESCC) and search the potential biomarkers of ESCC.
Methods:
Serum samples from 59 patients with ESCC and 57 healthy controls were collected, and randomly divided into the training group (44 patients and 42 healthy controls) and validation group (15 patients and 15 healthy controls). Serum proteins/peptides were extracted and purified with weak cation-exchange chromatography Magnetic Beads (WCX-MB), and detected by the matrix-assisted laser desorption / ionization time-of-flight mass spectrometry (MALDI-TOF MS). Then the differentially expressed proteins/peptides were screened out, and a diagnostic prediction model for ESCC was established and preliminarily validated.
Results:
The ClinProTools software identified 31 differential peptide peaks (P<0.05), among which 18 peaks had significant difference (P<0.01). Compared with healthy controls, 8 peaks were up-regulated in ESCC patients, while 10 peaks were down-regulated. Among them, the areas under the receiver operating characteristics (ROC) curve (AUC ROC ) of m/z 2 660.84 and m/z 5 336.49 peaks were 0.95 and 0.91, respectively, and their expressions were up-regulated in ESCC patients. The validation results showed that the accuracy, sensitivity and specificity of the diagnostic prediction model established by the genetic algorithm (GA) were 93.10%, 92.90% and 93.30%, respectively.
Conclusion
The established diagnostic prediction model may be used for the auxiliary diagnosis of ESCC. Two peptide peaks of m/z 2 660.84 and m/z 5 336.49 may be the potential biomarkers of ESCC.
2. Endothelial FGFR1 deficiency induces AcSDKP-resistant EndMT by regulating TGFβ signal pathway
Qiongying HU ; Chengjin AI ; Gaolin CHEN ; Daqian XIONG
Chinese Journal of Endocrinology and Metabolism 2019;35(11):969-972
Objective:
To investigate the role of fibroblast growth factor receptor(FGFR) 1 in endothelial to-mesenchymal transition(EndMT) and epithelial-to-mesenchymal transition(EMT), and to find out a new strategy to study the vascular endothelial function of diabetic renal fibrosis.
Methods:
Culture media from FRS2 knockdown HMVECs was transferred to HK-2 cells. Western blot and immunofluorescence staining were used to measure EMT markers and key moleculars of transforming growth factor(TGFβ).
Results:
It was found that the medium from FRS2 siRNA-transfected HMVECs reduced E-cadherin protein levels, increased EMT markers levels, and activated TGFβ signal pathway in HK-2 cells.
Conclusion
Endothelial FGFR1 deficiency-induced EndMT leads to EMT in neighboring cells in a manner dependent on TGFβ1 signaling. Endothelial cell FGFR1 is an important molecule for maintaining endothelial homeostasis and epithelial homeostasis, and seems to be a key target for anti-diabetic renal fibrosis.
3.Research of B7-H3 and carcinoembryonic antigen on diagnosing malignant pleural effusion
Qiongying HU ; Jianrong LUO ; Chengjin AI ; Gaoli CHEN ; Chaoming ZHANG ; Daqian XIONG
International Journal of Laboratory Medicine 2019;40(3):266-268
Objective To investigate the diagnostic value of B7-H3 and carcinoembryonic antigen (CEA) on diagnosis of malignant pleural effusion (MPE).Methods We collected and analysed the expression of B7-H3 and CEA in 40 MPE cases and 22 cases of benign pleural effusion (BPE) using enzyme-linked immunosorbent assay (ELISA).Receiver operator characteristic curves (ROC) were drawn according to the expression of B7-H3 and CEA, calculated diagnosis sensitivity, specificity and the area under curve (AUC).Results The diagnosis sensitivity of B7-H3 was 62.5%, specificity 81.0%, with the AUC of 0.777;similarly, The diagnosis sensitivity of CEA was 72.5%, specificity 81.0%, with the AUC of 0.850.Higher AUC of 0.910 was gained in combination ROC, with sensitivity and specificity of 72.5%, 81.0%, respectively.Conclusion B7-H3 and CEA could be available diagnosing markers for MPE.Combined applications of B7-H3 and CEA have higher AUC.They may be widely applied in future clinical practice.
4.The intestinal micro-ecology influences the clinical efficacy of immune checkpoint inhibitors in cancer patients
Ya'nan ZHOU ; Xiaotong ZHANG ; Chengjin HU ; Yuan CAO
Chinese Journal of Laboratory Medicine 2019;42(5):386-389
Immune checkpoint inhibitors (ICIs) targeting CTLA-4 and PD-1 /PD-L1 are increasingly used to treat several types of cancer.Although several agents have been approved,only 10% to 30% of patients have benefited from them.Some studies have shown that intestinal micro-ecology can affect the outcome of ICIs through immune regulation in cancer patients.This literature review s summarized the regulation and predictive functions of intestinal micro-ecology on ICIs treatment,and introduced possible mechanisms of intestinal micro-ecology that affacted the efficacy of immunotherapy.In addition,some approaches for detecting gut microbiome were also summarized.Gut microbiota highlighted in this review may serve as novel biomarkers to predict clinical outcomes in cancer patients.
5.Establishment of lung adenocarcinoma diagnosis and prediction model based on MALDI-TOF-MS technique and its preliminary verification
Xinyan JING ; Zhaolian LU ; Ting ZHANG ; Chengjin HU
International Journal of Laboratory Medicine 2018;39(9):1034-1037,1040
Objective To use the liquid protein combined with MALDI-TOF-MS for screening the serum differential peptides markers in lung adenocarcinoma patients and to establish the lung adenocarcinoma diag-nosed prediction model for founding the potential markers for the diagnosis of lung adenocarcinoma.Methods 37 patients with lung adenocarcinoma and 33 healthy subjects and benign lung disease which were made up in control group were collected,in the two groups the age and the sex were matched.The two groups were ran-domly divided into training group(30 cases of lung adenocarcinoma,26 cases of control)and test group(7 ca-ses of lung adenocarcinoma,7 cases of control)according to 3:1.T he differential diagnosis of lung adenocarci-noma and control group was performed by liquid chip-time-of-flight mass spectrometry and software ClinPro-Tools 3.0 to establish a prediction model of lung adenocarcinoma.The diagnostic model was validated by using serum samples from the test group to assess the diagnostic efficacy of the model.Results Nine peptide peaks with significant differences(P<0.05)were obtained by ClinProTools 3.0 software analysis.The up-regulated peaks in lung adenocarcinoma(m/z)were 8 976.5,4 469.05,4 966.78,8 925.5,4 531.05,and the down-reg-ulated m/z were 3 304.44,8 594.76,3 266.82,3 195.52.According to the genetic algorithm(GA),the lung ad-enocarcinoma diagnosis and prediction model was established.The overall recognition ability of the model was 94.49%.The model was evaluated by the test group.The results showed that the sensitivity of the model was 100.0% and the specificity was 85.7%.Conclusion Among lung adenocarcinoma patients,serum benign lung disease and healthy,there are differences in the serum peptide.T he use of differential peptide peaks to estab-lish lung adenocarcinoma diagnostic prediction model for the early diagnosis of lung adenocarcinoma provides a new method.
6.Bioinformatics Analysis of long non-coding RNA TUG1 in hepatocellular carcinoma
Yucui ZHU ; Xiaotong ZHANG ; Yanan ZHOU ; Chengjin HU ; Yuan CAO
International Journal of Laboratory Medicine 2018;39(10):1153-1157,1162
Objective To explore the significance of long non-coding RNA (lncRNA) taurine up-regulated gene 1 (TUG1) in hepatocellular carcinoma (HCC),to predict the target gene of TUG1,and to provide a ref-erence for further study of TUG1 in HCC.Methods The differential expression of TUG1 in HCC was ana-lyzed by using the UALCAN database and the survival analysis of TUG1 was performed.The target gene of TUG1 was predicted by RegRNA 2.0 biology software,HMDD,targetscan and microT-CDS,and the regulato-ry network of lncRNA TUG1-microRNAs-mRNAs was constructed.The predicted target gene was analyzed by Gene Ontology (GO) and KEGG signal transduction pathway enrichment by using FunRich platform. Results TUG1 expression in HCC was significantly increased,and the expression level of TUG1 increased generally with the increase of tumor grade.The overall survival of patients with low expression of lncRNA TUG1 was significantly longer than that of lncRNA TUG1 high expression patients.There were four possible binding sites of HCC related microRNAs (hsa-mir-122-5p,hsa-mir-200a-3p,hsa-mir-34c-3p,hsa-mir-629-3p) on TUG1,which regulated 245 downstream target genes and formed the regulatory network of lncRNA TUG1-microRNAs-mRNAs.In the biological process,microRNA target genes were highly enriched in the processes such as the regulation of nucleobase,nucleoside,nucleotide and nucleic acid metabolism.In KEGG pathway analysis,microRNA target genes were highly enriched to the signal pathways mediated by Syndecan and TRAIL.Conclusion TUG1 expression level in HCC increased.Increased expression of TUG1 is associat-ed with poor prognosis in HCC.Bioinformatics methods can be used to explore the mechanism of tumorigene-sis from the molecular level,which can provide valuable information for subsequent experiments and clinical diagnosis and treatment.
7.Changes in circulating microparticles in mice with ventilator-induced lung injury
Yingjie YIN ; Shouqin ZHANG ; Sanqiang WANG ; Hu LONG ; Chengjin GAO ; Xiangyu ZHANG
Chinese Critical Care Medicine 2018;30(1):8-12
Objective To investigate the changes in serum level of microparticles (EMPs) in mice with ventilator-induced lung injury (VILI), and explore its significance in VILI. Methods Forty-eight grade SPF male C57BL/6J mice were randomly divided into two groups, with 24 mice in each group: the mice in mechanical ventilation (MV) group were given high tidal volume (VT 30 mL/kg) MV for 4 hours after tracheal intubation, and those in spontaneous breathing group were spontaneously breathed for 4 hours. The apical blood of 12 mice in each group were collected, and serum levels of interleukins (IL-1β, IL-6) and tumor necrosis factor-α (TNF-α) were determined by enzyme linked immunosorbent assay (ELISA), and serum EMPs levels were determined by flow cytometer. The correlations between EMPs and IL-1β, IL-6, and TNF-α were analyzed by linear regression analysis. The lung tissues of other 12 mice in each group were harvested, and wet/dry weight (W/D) ratio was assessed. After hematoxylin-eosin (HE) staining, the morphological changes in lung tissue were observed under light microscope. After double staining of uranium acetate and lead citrate, the ultrastructural changes in lung tissue were observed with electron microscope. Results Compared with spontaneous breathing group, the levels of lung W/D ratio in MV group was significantly increased (5.47±0.14 vs. 4.34±0.11), the levels of IL-1β, IL-6, TNF-α and EMPs were also significantly increased [IL-1β (ng/L): 42.4±4.4 vs. 7.7±3.6, IL-6 (ng/L): 1 239.5±66.3 vs. 21.7±4.6, TNF-α (ng/L):237.6±25.8 vs. 37.1±19.1, EMPs (cells/μL): 28.6±1.8 vs. 5.9±1.8, all P < 0.01]. It was shown by correlation analysis that EMPs were positively related with IL-1β, IL-6, and TNF-α (r value was 0.968, 0.932, 0.945, respectively, all P = 0.000). It was shown by fitting linear regression analysis that when EMPs increased by 1 cell/μL, IL-1β increased by 2.4 ng/L [95% confidence interval (95%CI) = 1.9-2.8, P < 0.001], IL-6 increased by 34.5 ng/L (95%CI = 25.1-44.0, P < 0.001), and TNF-α increased by 13.6 ng/L (95%CI = 10.3-16.9,P < 0.001). It was shown by light microscope that the structure of lung tissue and alveolar of mice in spontaneous breathing group appeared normal, while the shrinks of alveolar and disappearance of alveolar architecture were found in MV group. It was shown by electron microscopy that alveolar wall edema and thickening and broken alveolar septa were found in MV group, by contrast, the structure of alveolar was normal in spontaneous breathing group. Conclusion 30 mL/kg VT ventilation for 4 hours could induce VILI with increase in EMPs, suggesting EMPs closely related to VILI, and EMPs level may be putative biomarker of VILI.
8.Clinical distribution and antimicrobial resistant characteristics of Pseudomonas aeruginosa in Northern Guangdong
Weiguo YIN ; Feijun ZHAO ; Helin ZHANG ; Jun LUO ; Tiebing ZENG ; Chengjin HU
International Journal of Laboratory Medicine 2017;38(1):21-23,26
Objective To investigate the clinical distribution and antimicrobial resistant characteristics of Pseudomonas aeruginosa(PA)in the northern area of Guangdong.Provide the reference for clinical to prevent infection and reasonable choice of antibiotics and reduce the production of drug resistance strains.Methods The separation and identification of PA were performed by conventional methods during the data of drug 2013 and 2014.The data of sensitivity test of PA were analyzed by WHONET 5.6 and SPSS19.0 softwares.Results The 584 strains PA were mainly distributed in ICU,department of orthopaedics and respiratory medicine.Specimens were mainly from sputum and wound secretion.The detection of PA to 12 antibacterial agents showed different resistance.The antimicrobial with highest resistance was the gentamicin and lower resistance rates to fluoroquinolones,carbapenems,enzyme inhibitors.And a downward trend was shown in drug resistance to CIP,FEP,LEV,SCF.Conclusion PA mainly cause lung and wound infection,especially those old patients that come from ICU,department of orthopaedics and respiratory medicine.Although the drug resistance rates of PA to the commonly used antibiotics are relatively low,The clinicians should reasonably use antibiotics so as to reduce the resistant strains,especially the produce of MDR-PA and PDR-PA.
9.Comparison of RPR′s performance in syphilis screening between pregnant women and non-pregnant women
Weiguo YIN ; Cuizhen LIU ; Helin ZHANG ; Jun LUO ; Xiaoyun HUANG ; Chengjin HU
International Journal of Laboratory Medicine 2017;38(10):1322-1323
Objective To study comparatively the performance of rapid plasma reagin (RPR) test in syphilis detection among pregnant women and non-pregnant women to provide reference for detecting syphilis in pregnant women.Methods The women aged 20-40 years old were selected and divided into the pregnant group and the non-pregnant group.RPR and treponema pallidum particle assay(TPPA) were simultaneously adopted to conduct the syphilis detection.The positive cases were judged by the TPPA detection results combined with the contact history,clinical symptoms and treatment situation.The results were compared with those by RPR for determining the false negative and false positive in RPR.The false negative rate and false positive rate of RPR detection results were analyzed in the two groups.Results Among 117 pregnant women,15 cases were false negative in RPR and 9 cases were false positive in RPR;among 755 non-pregnant women,there were 44 cases of false negative RPR results and 8 cases of false positive RPR results.The false negative rates in the pregnant group and non-pregnant group were 25.0% and 8.8% respectively,the difference was statistically significant(χ2=14.739,P<0.05);the false positive rates in the pregnant group and non-pregnant group were 15.7% and 3.1% respectively,the difference was statistically significant (χ2=14.722,P<0.05).Conclusion There are many factors affecting RPR for detection syphilis,pregnant women are the specific group,so higher false positive rate and false negative rate exist than non-pregnant women,the detection results should be comprehensively judged by combining with clinical symptoms and disease history,if necessary,combining with other syphilis detection method for avoiding missed diagnosis and misdiagnosis.
10.Development and clinical application of a Sandwich ELISA for hK6 detection
Hua ZHANG ; Chengjin HU ; Kuixiang CHEN
International Journal of Laboratory Medicine 2017;38(17):2337-2338,2341
Objective To prepare the mAbs against hK6 for establishing a sandwich enzyme-linked immunosorbent assay(ELISA) of hK6,and exploring its clinical value.Methods The hybridoma technique was used to prepare mAbs against hK6.The mAbs were purified and labelled with horseradish peroxidase for the sandwich ELISA method.The sandwich ELISA method was used to detect the serum hK6 concentrations in patients with malignant gastric neoplasm.Then the best antibody pair was selected from coating antibody and enzyme-linked antibody to establish a sandwich ELISA method through the chessboard titrations.Compared with CEA,we explored the feasibility of hK6 as gastric cancer biomarkers.Results A sandwich ELISA method was established for quantifying hK6 in serum.The results showed that the optimal concentration of coating antibody was 5 μg/mL.The optimal concentration of enzyme-linked antibody was 1:2 000.Serum hK6 in the patients with gastric cancer groups[(5.78±1.66)ng/mL] than healthy individuals groups[(3.35±0.67)ng/mL] and those in gastric ulcer groups[(3.59±1.02)ng/mL],the difference was statistically significant(P<0.05).Furthermore,there was no significant difference in values of serum hK6 between patients with gastric ulcer groups and healthy individuals groups(P>0.05).The hK6 positive rate of gastric cancer was 69.70%,and CEA was 45.46%.In the combined detection,the positive rate was 78.79%.Conclusion A sandwich ELISA is established successfully.As a favorable serum biomarker for gastric cancer,the detection of hK6 together with CEA is helpful in the diagnosis of gastric cancer.

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