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Objective:To find the age-related indicators of human immune function in the blood, to explore the possibility to standardize the evaluation criterion of immune function in Chinese population.Methods:The peripheral blood samples from 478 healthy individuals with complete medical data collected at the Beijing Hospital from January 2019 to December 2019, were chosen and analyzed.The volunteers were divided into young(<40 years old)and elderly(≥70 years old)groups.The differences in RNA, cytokines and immune cell function(the proliferation of T cell as well as the ability of T cells and natural killer cells to lyse target cells)were compared between the young group versus the elderly group.Results:Real-time quantitative fluorescence RT-PCR detection method was used for detecting the following tumor immune-related genes in the peripheral blood of healthy people in young versus old groups: programmed death 1( PD1)gene, complement C3 gene( C3), complement C4 gene( C4), high sensitivity C-reactive protein gene( HsCRP), tumor necrosis factor alpha gene( TNFA), interferon gamma gene( INF- γ), interleukin-6 gene( IL-6), interleukin-8 gene( IL-8), cytotoxic T lymphocyte-associated antigen-4 gene( CTLA-4), nuclear transcription factor-κB gene( NF- κB), long intergenic noncoding RNA-erythroid pro-survival gene( lincRNA- EPS), long non-coding RNA-myeloid RNA regulator of Bim-induced death gene( lncRNA- Morrbid), lincRNA-cyclooxygenase 2 gene( lincRNA- Cox2), Lnc-Dendritic cells( Lnc- DC), and circRNA-7 gene( ciRS-7). The mRNA level of C4 was significantly lower in elderly people than in young controls[young group(1.01±0.18) vs.old group(0.64±0.13), P=0.047], while others showing a non-statistically significant difference( P>0.05). The protein levels of programmed death 1(PD1), complement C3(C3), complement C4(C4), high sensitivity C-reactive protein(HsCRP), tumor necrosis factor alpha(TNF-α), interferon gamma(INF-γ), interleukin-6(IL-6), interleukin-8(IL-8), cytotoxic T lymphocyte-associated antigen-4(CTLA-4)and nuclear transcription factor-κB(NF-κB)in plasma were detected by enzyme-linked immunosorbent assay(ELISA). The following protein levels were significantly higher in the elderly group than in the young group: IL-6 protein[young group(249.30±100.02)ng/L vs.old group(283.00±94.98)ng/L, P=0.021], HsCRP protein[young group(2543.00±1111.89)ng/L vs.old group(3056.00±1056.61)ng/L, P=0.002], INF-γ protein[young group(362.40±383.67)ng/L vs.old group(500.40±502.27)ng/L, P=0.047]and TNF-α protein[young group(20.74±29.47)ng/L vs.old group(33.09±48.91)ng/L, P=0.042]. While the level of C4 was significantly lower in the elderly group than in the young group[young group(449.50±51.17)ng/L vs.old group(407.10±59.78)ng/L, P<0.0001]. T-cell proliferation, quantified by flow cytometry-based fluorescent dye dilution, showed that the CD3 + T cells of elder people had proliferated through 4 generations, while the CD3 + T cells of young people had proliferated through 5 generations.As shown in the NK killing assays, the secretion of LDH in the target A549 tumor cells decreased significantly after treated with NK cells belonging to elder individuals.The result demonstrated that individuals over 70 years old have significantly lower levels of the killing activity of NK cells( P<0.05). However, there was not statistically significant in T lymphocyte killing activity between different ages( P>0.05). Conclusions:IL-6, HsCRP, INF-γ and TNF-α levels are increased with aging, while the level of C4 significantly is decreased.The proliferation ability of T cells and the ability of NK cells to kill tumor cells are weakened.
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Objective In order to establish a basis for exploring diagnostic biomarkers of type 2 diabetes mellitus (T2DM), our research screened differentially expressed circRNAs in high glucose treated human umbilical vein endothelial cells ( HUVECs) and validated these circRNAs in both cell models and peripheral blood samples of T 2DM patients.Methods The research used HUVECs as experimental model . The control group (n=3) and high glucose group (n=3) were set up and treated under normal condition (5.5 mmol/L) and high glucose condition (30.0 mmol/L), respectively.The research utilized high-throughput sequencing technology to preliminarily screen differentially expressed circRNAs .Differentially expressed circular RNAs were validated in the endothelial cell model by PCR and real -time quantitative PCR ( q-PCR) techniques.Subsequently , in the peripheral blood samples of T 2DM patients ( n=32) and control individuals ( n=28 ), the differentially expressed circRNAs were further validated . Finally, t-Test, correlation analysis and ROC curve were used to analyze the experimental results .Results Total 1087 differentially expressed circular RNAs were screened by high-throughput sequencing;and of these , 554 were up-regulated, 533 were down-regulated.Six increased circRNAs were selected and validated in HUVEC model;and their PCR and q-PCR results matched with sequencing results .Further validation was conducted in peripheral blood samples for three most up-regulated circRNAs.This study found that the differential expression of hsa_circ_0031739 in peripheral blood samples was statistically significant ( 0.015 ±0.0025 vs.0.006 ±0.0013,P=0.0059 <0.05).The area under the ROC curve (AUC) was 0.730 and the expression level of hsa_circ_0031739 was positively correlated with blood glucose ( GLU ) and glycated hemoglobin ( GHb ) ( r=0.317, P=0.0137 <0.05;r=0.348, P=0.0064 <0.05 ) .Conclusion HUVECs were commonly utilized as cell models in the study of T 2DM, and differentially expressed circular RNA profiles existed afterhigh glucose treatment .The differential expression of hsa_circ_0031739 was significant in both high glucose treated HUVECs and peripheral blood samples of T 2DM patients, and the expression level of hsa_circ_0031739 was correlated with GLU and GHb , which has certain diagnostic significance.Therefore, hsa_circ_0031739 may become a new diagnostic biomarker for T2DM and be helpful for the comprehensive diagnosis and mechanism research of T 2DM.
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Currently, clinical laboratories are developing towards two different directions .One is centralized laboratory with automated pipelines , and the other is highly integrated portable detection system.The latter evolved from an early single test device gradually to an integrated platform based on microfluidic technology , which can operate multiple steps and tests simultaneously .The microfluidic system is further developed to a miniaturized device which can perform biochemical , immune, microbial and cytological tests.With the medical reform process in China , the microfluidic systems will appear in the primary health unit or even families .This review concludes the current application of microfluidic technology in clinical diagnostics and its future trends .
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Objectives To analyze the relationship between phosphorylation of serine-3 of cofilin1 and Taxol resistance of ovarian cancer and to evaluate the prognostic value of cofilin1 phosphorylation in estimating ovarian cancer survival using clinical samples.Methods Wild type (WT) plasmids with over-expression of wild cofilin1,S3L plasmids with over-expression of cofilin1 and inhibited phosphorylation at serine-3,and siRNA-C plasmids with down-regulation of cofilin1 were constructed using molecular biology techniques.After the SKOV3 and SK-TR30 cell-lines were transfected with these plasmids,changes in biological characteristics and drug resistance after instant transfection were compared.Fifty-one elderly female patients with microscopically confirmed ovarian cancer,who had received chemotherapy with Taxol and cis-platinum (TC)after surgery,including 30 chemo-sensitive and 21 chemo-resistant cases,were recruited in this study.Immunohistochemical methods were used to detect the expression of cofilinl and phosphorylated cofilin in tissue sections from the two groups.Progression free survival(PFS)was analyzed.Results Compared with the control group,the proportion of cells increased at the G0/G1 phase(P=0.034)and decreased at the S phase (P=0.031),and the apoptosis rate decreased(P=0.020),in SKOV3 cells with high expression of cofilin1.However,these characteristics disappeared when the wild type was replaced with the inactive mutant S3L.When cofilin1 expression was down-regulated in SK-TR30 cells,the proportion of cells at the G0/G1 phase decreased(P=0.020).The expression of cofilin1 was detected in 56.7 ％ (17/30)and 57.1％ (12/21),respectively,sections of ovarian tumor tissues of the chemo-sensitive and chemoresistant groups,and there was no statistic difference (x2=0.332,P =0.943)between the two groups.The expression of phosphorylated cofilin was much higher in the chemo-resistant group (85.7％ or 18/21)than in the chemo-sensitive group (53.3％ or 16/30),and the difference was statistically significant(x2=5.829,P=0.016).The higher expression of phosphorylated cofilin was also correlated with shorter PFS(x2 =21.440,P＜0.01,95％ CI:0.068-0.883),especially in the chemo-sensitive group.Conclusions Serine-3 phosphorylation status of cofilin 1 is associated with paclitaxel resistance in ovarian cancer,but the underlying mechanisms of regulation need further investigation.
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AP-2,a nuclear transcription factor family,always binds to a typical DNA sequence in the dimer way,thus AP-2 can regulates the expression of many other genes and participate in the growth and devel-opment, apoptosis and many other physiological processes. Under pathological conditions, AP-2 also plays an im-portant role in tumor genesis and development by influencing cell proliferation, differentiation, cell cycle regula-tion and apoptosis. Recent years,many researchers want to find new therapeutic strategies that target AP-2 in a series of human cancers.