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Objective:To detect the concentration of various cytokines in aqueous humor of patients with diabetes retinopathy (DR) with Luminex liquid chip, and analyze the relationship between the cytokines and the occurrence and development of DR.Methods:A cross-sectional study was conducted.Sixty-three DR patients (97 eyes) treated with anti-vascular endothelial growth factor (VEGF) drugs in the First Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine from March 1, 2019 to December 31, 2019 were enrolled as DR group, including 38 nonproliferative DR (NPDR) eyes in NPDR group and 59 proliferative DR (PDR) eyes in PDR group, 39 eyes in photocoagulation group and 58 eyes in non-photocoagulation group.Twenty-seven patients (31 eyes) hospitalized for cataract surgery at the same time were collected as the control group.Aqueous humor was extracted during the operation, and Luminex liquid chip was used to detect the concentrations of vascular endothelial growth factor-A (VEGF-A), placental growth factor (PLGF), platelet-derived growth factor (PDGF)-AA, PDGF-BB, angiopoietin-like protein 4 (ANGPTL4), interleukin-6 (IL-6), IL-8, IL-1β, monocyte chemotactic protein-1 (MCP-1), intercellular cell adhesion molecule-1 (ICAM-1) and tumor necrosis factor-α (TNF-α) in aqueous humor.The concentrations of various cytokines of different groups were compared, and the correlation among various aqueous humor cytokines was analyzed by Spearman rank correlation analysis.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of First Affiliated Hospital of Guangzhou University of Chinese Medicine (No.Y[2019]230). Written informed consent was obtained from each patient.Results:The concentrations of VEGF-A, PLGF, PDGF-AA, ANGPTL4, IL-6, IL-8, MCP-1 and ICAM-1 in DR group were significantly higher and the concentration of IL-1β was significantly lower than those of control group ( Z=-4.747, -5.164, -3.373, -8.062, -4.535, -5.954, -5.098, -3.228, -5.954, all at P<0.01). The concentrations of VEGF-A, PLGF, PDGF-AA, ANGPTL4, IL-6, IL-8 and MCP-1 of the photocoagulation and non-photocoagulation groups were higher and the concentration of IL-1β was significantly lower than those in the control group (all at P<0.017). The concentration of ICAM-1 in the photocoagulation group was significantly higher than that in the control group ( P<0.017). The concentrations of PLGF, PDGF-AA and ANGPTL4 of PDR group were higher than those of NPDR group, and the differences were statistically significant ( Z=-2.291, -3.396, -2.276, all at P<0.05). VEGF-A was positively correlated with the other cytokines except ICAM-1 ( rs=0.237-0.540, all at P<0.05). ANGPTL4 was positively correlated with the other cytokines except IL-1β ( rs=0.361-0.733, all at P<0.01). Conclusions:The occurrence and development of DR are closely related to VEGF family, PDGF family, ANGPTL family and inflammatory factors.The concentrations of PLGF, PDGF-AA and ANGPTL4 are higher in PDR eyes.There are close and complex relationships among a variety of cytokines in the eyes of DR patients.
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Objective @#To examine the effect of asbestos exposure on oxidative stress, so as to provide insights into the elucidation of pathogenesis and management of asbestos-related diseases. @*Methods @#Totally 245 subjects were recruited from an asbestos manufacturing area in Zhejiang Province, and their gender, age and history of asbestos exposure were collected through a questionnaire survey. The serum levels of 8-hydroxy-2'deoxyguanosine ( 8-OHdG ), glutathione ( GSH ), malondialdehyde ( MDA ), superoxide dismutase ( SOD ) and total antioxidative capacity ( TAOC ) were measured using an enzyme-linked immunosorbent assay ( ELISA ), and the levels of catalase ( CAT ), peroxiredoxin 2 ( PRX2 ), SOD1, SOD2 and thioredoxin-1 ( TRX1 ) were detected in peripheral white blood cells ( WBCs ) using a liquid-chip assay. Multivariable linear regression analysis was performed to identify the association between asbestos exposure and oxidative stress parameters. @*Results @#There were 50 subjects without a history of asbestos exposure (unexposed group), 102 subjects with asbestos exposure for less than 10 years ( AE<10-year group ) and 93 subjects with asbestos exposure for 10 years and more ( AE≥10-year group ). No significant differences were found among the three groups in terms of age, gender, proportion of smokers or proportion of alcohol consumers ( P>0.05 ). Significantly higher 8-OHdG and MDA in serum, and higher PRX2 in peripheral WBCs were detected in the AE≥10-year group than in the unexposed group ( P<0.05 ); lower GSH and TAOC in serum, and lower CAT in peripheral WBCs were detected in the AE≥10-year group than in the unexposed group ( P<0.05 ); higher 8-OHdG and MDA in serum, and higher PRX2 in peripheral WBCs were detected in the AE≥10-year group than in the AE<10-year group ( P<0.05 ). Multivariable linear regression analysis showed that asbestos exposure significantly correlated with 8-OHdG, MDA and TAOC in serum, and CAT and PRX2 in peripheral WBCs ( P<0.05 ). @*Conclusion @#Asbestos exposure may induce the oxidative stress damage, suggesting that oxidative stress may be involved in asbestos-related diseases.
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Objective:To evaluate the diagnostic capabilities of PCR-flow Fluorescence Hybridization technology in prenatal genetic diagnosis of thalassemia.Methods:8 005 cases of prenatal genetic diagnosis of thalassemia in Guangdong Women and Children Hospital from September 2017 to December 2020 were retrospectively analyzed. All samples were diagnosed by traditional genetic methods include multiple Gap-PCR, PCR-RDB, MLPA and Sanger sequencing. Meanwhile, PCR-flow Fluorescence Hybridization technology was used as a verification platform for detecting common mutation sites of thalassemia. The results were analyzed to evaluate the diagnostic capabilities of PCR-flow Fluorescence Hybridization technology compared with traditional methods in prenatal genetic diagnosis of thalassemia.Results:By traditional methods, 1 939 cases (24.22%, 1 939/8 005) were normal and 6 066 cases (75.78%, 6 066/8 005) were diagnosed as thalassemia, including 4 513 cases of α-thalassemia, 1 475 cases of β-thalassemia, and 78 cases of αβ-thalassemia. By PCR-flow Fluorescence Hybridization technology, 7 845 samples were successfully diagnosed after initial interpretation by software. Compared with traditional methods, all the sensitivity, specificity and accuracy were 100%. The other 160 samples which failed in the initial interpretation can be successfully interpreted after review or manual interpretation.Conclusion:There were no differences between the two methods on the detecting of common mutation sites of thalassemia.
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@#Objective: To investigate the effects of XIAOJI Decoction combined with FOLFOX chemotherapy on serum cytokine expression profile in patients with advanced colorectal carcinoma by liquid chip technology. Methods: Fourteen patients with advanced colorectal carcinoma, who met the inclusion criteria and were treated in the Department of Oncology, Higher Education Mega Center Hospital of Guangdong Provincial Hospital of Traditional Chinese Medicine during January 1, 2018 and December 31, 2018 were retrospectively analyzed in this study. The patients were divided into chemotherapy group (n=7, treated with 5-Fluorouracil + Calcium Folic Acid+Oxaliplatin (FOLFOX)) and combined treatment group (n=7, treated with XIAOJI Decoction + FOLFOX) according to therapeutic scheme. The curative efficacy was evaluated after 6 treatment courses. The expression profile of cytokines in blood serum of patients was examined by liquid chip technology after every 2 courses. Results: Fourteen patients received a total of 84 cycles of therapy. Survival analyses showed that the progress-free survival time (PFS) and overall survival time (OS) of two groups couldn't be compared due to insufficient samples, although the combined treatment group had longer PFS (10 months vs 6 months) and OS (17 months vs 12 months) than the chemotherapy group.As to adverse reactions, the rates of leucopenia, diarrhea, nausea, peripheral neuritis and alopecia in two groups were comparable, while the severity in combined treatment group were lighter than that in chemotherapy group. In comparison with the combined treatment group, concentrations of serum BDNF and IL-2 were statistically higher in the chemotherapy group (P<0.05). By comparing the cytokine concentrations at different collection time points before and after the treatment, it showed that the concentration of serum IL-2 in chemotherapy group was higher than that in combined treatment group after 2 courses of treatment (P<0.05). In total, there were 19 cytokines showed a tendency to be higher in combined treatment group than chemotherapy group during different treatment periods. Conclusion: Combined treatment of XIAOJI Decoction with FOLFOX for advanced colorectal carcinoma is a treatment option worth exploring, and liquid chip analysis showed that the mechanism may be related to the reduction of serum LI-2 and BDNF levels in patients.
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Objective@#To establish a quantitative assay of serum Golgi protein 73 (GP73) using xMAP technology and evaluate its performance.@*Methods@#Monoclonal antibodies against GP73 were prepared and purified, and antibody pair screening was performed by double-antibody sandwich enzyme-linked immunosorbent assay. The screened antibodies were used to construct a Luminex liquid chip detection system, and the analysis performance of the detection system was evaluated. The serum levels of GP73 were detected in 90 clinical samples from healthy controls and patients with chronic hepatitis B infection (CHB) and hepatocellular carcinoma (HCC).@*Results@#Five anti-GP73 monoclonal antibodies were prepared and purified, and 5 antibody pairs were successfully screened. The Luminex liquid chip detection system of GP73 was successfully constructed using 8F10D1 and 10B9F11 antibody pairs. The analytical performance evaluation showed that the sensitivity of this system was 0.25 ng/ml and the dynamic range was 0.25-100 ng/ml. No cross reactivity was observed. The intra- and inter-assay variation for GP73 was <8% and <11%, respectively. The recovery was 83%-92%. The linear regression equation was y=1.141x+ 6.436 (r2=0.998 4, P<0.001). The GP73 concentrations in the serum samples of healthy control, CHB group, and HCC group were 42.8 (38.68, 55.90) ng/ml, 61.49 (43.59, 81) ng/ml, and 122.78 (49.36 liter, 264.55) ng/ml, respectively. The levels of GP73 in HCC group were significantly higher than those in CHB group and healthy controls (P<0.05). Moreover, the levels of GP73 in CHB group were significantly higher than those in healthy controls (P<0.05).@*Conclusions@#A liquid chip detection system of GP73 was successfully constructed. It provides a powerful tool for the clinical application of GP73 in the future.
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Objective This study aimed to evaluate the clinical significance of detecting cytokine expression with Crohn's disease(CD) by Luminex liquid chip.Methods A total of 76 patients with CD and 50 healthy volunteers as healthy control group were recruited in this study.The expression of cytokines IL-2,IFN-γ,TNF-α,IL-4,IL-6,IL-10 and IL-17A were detected by Luminex liquid chip according to the instruction of MILLIPLEX MAP Kit.Enzyme-linked immunosorbent assay(ELISA) was used to detect the levels of anti-saccharomyces cerevisiae antibodies(ASCA).Then the relationship and differences among these groups were analyzed.Results Except IL-17A,the level of IL-2,IFN-γ,TNF-α,IL-4,IL-6,IL-10 was higher in the CD group than those in control group.The serum levels of IFN-γ,TNF-α and IL-6 in patients with active stage of CD group were significantly higher than those with remission stage and healthy control group.The level of TNF-α and IL-6 in moderate and severe stages were higher than those in slight stage.The serum level of ASCA had significant difference between CD group and control group.Compared with ASCA negative group,the ASCA positive patients have a higher level of IL-6 and TNF-α.ConclusionThe cytokines like IL-6 and TNF-α is correlated with CD severity,which is important for inflammatory activity and progression of CD.Altogether,these results demonstrated that the dynamic change of cytokines had a clear relativity in stage of CD,which is very imperative for diagnosis and clinical appraisal of CD.
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Liquid chip technology have been licensed to be used in clinic because of its advantage of high-throughput, high-sensitivity, good signal to noise ratio, reaction in liquid phase, convenient operation and short time consuming, etc. The optimization of a liquid chip system for the detection of serum biomarkers of colorectal tumour and initial application in the detection of CEA were studied. The optimized reaction conditions of liquid chip were determined through orthogonal design after it was prepared. The results showed that the consuming reaction time of the coated antibody and the antigen was 1hour. The microspheres, biotinylated detecion antibody and the consuming complexes and avidin-PE time of the microspheres and the biotinylated tested antibody was 1hour, 1hour and 15minutes respectively.the consuming time of the complexes and avidin-PE was fifteen minutes, The optimized dilution of the biotinylated tested detection antibody was 1∶300 and the optimized concentration of avidin-PE was 12?g/ml. Totally 55 clinical samples were detected by the liquid chip and by Enzyme-Linked Immunosorbent Assay (ELISA) simultaneously and the results of the two methods were compared. The results of the two methods showed good correlation between positive and negative samples but the detection limits and the dynamic ranges of the liquid chip method were more sensitive and wider than those of the ELISA. The multiple tumour biomarkers may be detected simultaneously and the time of clinical test and manpower requirements were reduced by the liquid chip method.
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Liquid chip,a new type of biochip classify encoding beads as carriers of reaction and signal detection,implement simultaneous determination protein,nucleic acid and other biomacromolecule in susp-ension liquid system.Apart from high-throughput characteristic as traditional biochip,the advantages of liquid chip are easy operation,the high-sensitivity,the high-accuracy,the high-precision and the great dynamic range and so on.It can be applied in the field of clinical laboratory diagnosis.In this review,we discuss the fundamental principle,detection,characteristics of liquid chip technique and the application in the field of clinical laboratory diagnosis.