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Objective:This work aims to assess the distribution of peripheral blood monocyte subsets, the expression level of the functional markers in rheumatoid arthritis (RA) patients, and analyze the correlation between the above indexes and the onset of RA.Methods:Peripheral blood mononuclear cells were collected and isolated from 62 RA patients, 52 healthy control (HC) and 12 disease control group′s patients via density centrifugation. The enrolled patients were attended or underwent physical examination in East Hospital, Tongji University from June 2020 to December 2021. Monocytes could be classified into classical (CM), intermediate (IM) and non-classical (NCM). Then, the flow cytometry was performed to examine the distribution of monocyte subsets and the measure the expression level of human leukocyte antigen DR (HLA-DR), intracellular tumor necrosis factor α (TNF-α) in peripheral blood monocytes. The statistical methods in this study mainly include: Kruskal-Wallis H test, Chi-Square test, Mann-Whitney U test, Wilcoxon matched-pairs signed ranks test, Spearman correlation coefficient test and Logistic regression analysis. The diagnostic value of IM proportion in RA was analyzed by ROC curve. Results:The monocytes number and monocytes proportion in white blood cells were much higher in RA [0.40 (0.40, 0.50), 7.60% (5.97%, 8.53%)] and disease control [0.40 (0.40, 0.68), 8.20% (5.85%, 10.28%)] compared with HC [0.30 (0.30, 0.40), 5.80% (5.03%, 6.38%)] ( H=24.733, P<0.001; H=27.469, P<0.001). A statistic-significant difference was detected among the proportion of CM[85.49%(76.91%,89.21%),88.94%(86.36%,91.72%),90.26%(80.25%, 92.56%)],IM[11.65%(8.47%,17.89%),7.89%(5.36%,10.75%), 5.56%(4.17%, 8.27%)], NCM[2.22%(1.39%, 3.74%), 2.49%(1.74%, 4.66%), 5.13%(3.39%, 9.85%)] in RA group, HC group and disease control group ( H=11.389, P=0.003; H=20.815, P<0.001; H=10.640, P=0.005). The proportion of CM was lower in RA and the IM proportion was increased in RA( P=0.003; P=0.003). The intracellular TNF-α level of monocytes in all three groups revealed the trend that IM>NCM>CM. The intracellular TNF-α in IM of RA was positively associated with serum TNF-α ( r=0.376, P=0.041). The HLA-DR expression in IM subsets were higher than CM and NCM subsets in all RA,HC and disease control groups. The expression of HLA-DR of IM in RA group and disease control was higher than HC group [8 611.50 (6201.3, 9890.8), 10 295.0 (7 899.0, 13632.0), 6 278.00(4 057.8, 9522.0), H=10.495, P=0.005]. There were no correlations between the proportion of peripheral blood IM and clinical characteristics CRP ( r=0.119, P=0.359), RF ( r=0.204, P=0.112) and ESR ( r=0.153, P=0.236). Logistic regression analysis showed that the proportion of IM ( OR=1.169, 95% CI 1.003-1.363, P=0.046), CRP ( OR=1.277, 95% CI 1.000-1.631, P=0.050), RF ( OR=1.179, 95% CI 1.080-1.287, P<0.001) are positively correlated with RA onset. The area under ROC curve for diagnosis of RA with IM proportion was 0.687, and the 95% confidence interval was 0.590-0.784, P<0.001. Conclusions:The distribution of monocyte subsets in peripheral blood of RA patients is abnormal. The increase in the proportion of IM, the enhanced antigen-presenting ability, and the increased level of TNF-α secretion in RA patients may play an important role in the pathogenesis of RA.
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Rheumatoid arthritis (RA) is a chronic erosive arthritis. Early diagnosis, standardized treatment and regular monitoring of the disease will effectively mitigate disease progression and reduce the disability rate. Currently, traditional synthetic disease-modifying antirheumatic drugs (DMARDs) are used alone or in combination with new biological DMARDs or targeted synthetic DMARDS in the treatment of RA, resulting in effective remission in some refractory patients. However, the efficacy and toxicities of different treatments varies. With the development of proteomic and epigenetic technologies, some proteins, non-coding RNAs, and anti-drug antibodies (ADA) have been identified as potential markers for early diagnosis, concomitant diagnosis and disease assessment of RA. We summarized and analyzed the application prospects of novel RA diagnosis markers, including serum proteins, cell membrane proteins, non-coding RNAs, and ADA, with the aim of promoting the application of new markers that allow more precise diagnosis and treatment of RA.
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Objective:To explore the clinical significance of combined detection of the promoter methylation of plasma free Septin9, SDC2 and BCAT1 genes in peripheral blood for the diagnosis of colorectal cancer. Methods The data of patients admitted to the Department of Gastroenterology, Shanghai East Hospital Affiliated to Tongji University from January to September 2019 were retrospectively analyzed. They were divided into colorectal cancer group (62 cases of colon cancer, 59 cases of rectal cancer), precancerous lesions group (77 cases of colorectal adenoma, 5 cases of high-grade intraepithelial neoplasia), interference group (61 cases of colorectal cancer and advanced adenoma negative but suffered other intestinal lesions, 17 cases of non-colorectal cancer) and healthy group (94 cases). The methylation status of three genes (Septin9, SDC2 and BCAT1) in peripheral blood plasma was detected simultaneously by fluorescence PCR. The relationship between the positive rate of three genes detected jointly and the clinic pathological characteristics of colorectal cancer was analyzed and compared with serum carcinoembryonic antigen (CEA) positive rate. The colorectal cancer group was divided into stage Ⅰ, Ⅱ, Ⅲ and Ⅳ according to TNM stage, and the colorectal cancer group was analyzed and counted by grade. The diagnostic efficiency of detection methods was analyzed by receiver operating characteristic (ROC) curve, and the area under ROC curve (AUC) was compared.Results:The positive rate of combined detection of SDC2 and BCAT1 gene methylation was higher than other three groups (χ 2 =237.246, P<0.001). The positive rate of combined detection of plasma Septin9, SDC2 and BCAT1 gene methylation was higher than CEA in colorectal cancer group ( P<0.001). The positive rates of the combined detection of plasma Septin9, SDC2 and BCAT1 gene methylation in stage Ⅰ-Ⅳ of colorectal cancer group were 73%(16/22), 87%(34/39), 86%(30/35) and 96%(24/25), respectively. Compared with CEA group, the positive rate of combined detection of plasma Septin9, SDC2 and BCAT1 gene methylation in stage Ⅰ-Ⅲ of colorectal cancer group was higher than serum CEA ( P<0.001), but the positive rate of stage Ⅳ was not statistically significant compared with CEA group ( P>0.05). ROC curve analysis showed that the AUC of Septin9, SDC2 and BCAT1 was 0.857(95% CI 0.810-0.903),0.819(95% CI 0.768-0.871)and 0.862(95% CI 0.816-0.909), respectively. The AUC of combined detection of three gene methylations was 0.889 (95% CI 0.846-0.933), and the AUC of combined detection with serum CEA was 0.913 (95% CI 0.874-0.951). There was no significant difference in the positive rate of combined detection of Septin9, SDC2 and BCAT1 gene methylation among different gender, age and cancerous site of colon cancer patients (all P>0.05). Conclusion:The combined detection of the promoter methylation of plasma free Septin9, SDC2 and BCAT1 genes in peripheral blood plasma is helpful for the early diagnosis of colorectal cancer. The positive rate in stage Ⅰ-Ⅲ of colorectal cancer group is higher than serum CEA. The combined diagnosis of the three genes can improve the diagnostic efficiency.
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Immunoglobulin G (IgG) is the main immunoglobulin in human serum and can be divided into four subclasses of IgGl, IgG2, IgG3and IgG4, respectively. IgG mainly play protective roles in body immunity. The structures of IgG subclass are different, therefore, their functions ale also different in the occurrence of diseases. There is evidence that IgG subclass analysis has important clinical application value in the diagnosis, pathogenesis and prognosis of IgG4-related diseases, antibody deficiency and other diseases. Accelerating development and transformation of new technologies for IgG subclass detection, focusing on IgG subclass detection and clinical applications will be helpful to improve the ability to diagnose and treat the difficult and complex diseases.
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Objective To establish an ultra high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method for detecting α-hydroxybutyrate (α-HB) in serum.Methods Electrospray ionization negative ion and multiple reaction monitoring mode were used to detect serum α-HB.The linearity,low limits of quantification,precision,recovery and interference of UHPLC-MS/MS were evaluated.The reference interval of this method was established in 130 serum samples (62 males and 68 females) from Shanghai East Hospital.Dixon method was used to judge the outliers and K-S test was used to analyze the data normality.The standard curve was scored by linear regression analysis.Results The total run time was 4 min of UHPLC-MS/MS method for the determination of α-HB.It has a good linear relationship in the range of 0.5-40.0 mg/L(r=0.999 4);the low limit of quantification was 0.5 mg/L;the in-batch and inter-batch coefficient of variation precision were less than 4.1％ and 6.3％,respectively;the recovery ranged between 95.8％ and 103.8％.Hemolytic samples (about 5 g/L hemoglobin),lipemic samples (about 12 mmol/L triglyceride),icteric samples (about 150 μmol/L total bilirubin) had no significant interference to the detection.The reference range of the apparent healthy population was 1.46-6.48 mg/L.Conclusions A method for the determination of serum α-HB by UHPLC-MS/MS was established.The method was simple,rapid,and could be used for the detection of clinical samples.
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Objective@#To establish a high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method for the detection of serum oleic acid (OA), and preliminarily evaluate the role of OA in insulin resistance (IR) of type 2 diabetes (T2DM). @*Methods@#OA-［ 13 C 5 ］ was used as isotope-labeled internal standard, and the ion pairs of OA and OA-［ 13 C 5 ］ were 281.3/281.3 and 286.3/286.3, respectively. The ultrapure water was used as mobile phase A and methanol: acetonitrile (1∶1, v/v) as mobile phase B in a ZORBAX SB-Aq C18 reversed phase column. Meanwhile, the gradient elution system with a flow rate of 0.3 mL/min was used. According to the CLSI guidelines (EP15-A3), the reliability of the established method was evaluated by detecting the performance indicators such as precision, trueness, linear range, stability and carrying contamination rate. Serum OA levels were detected by the established HPLC-MS/MS method in 109 patients with clinically diagnosed T2DM and 100 healthy controls. The insulin resistance index (HOMA-IR) was calculated to evaluate IR, and the relationship between OA and IR was further analyzed. @*Results@#The established HPLC-MS/MS method for the detection of serum OA had good specificity and linearity in the range of 10-1 000 μmol/L (y=0.007 55x+0.004 83,r=0.997 7), and the low limit of quantification (LLOQ) was 10 μmol/L. It also had good precision, and the within-run coefficient of variation (CV) and total CV were not more than 1.62% and 1.73%, respectively, indicating that the method was suitable for the detection of serum OA. The serum OA levels in T2DM patients ［(425.58 ± 220.17) μmol/L］ were significantly higher than that in the healthy controls ［(113.20±58.00) μmol/L］, and serum OA levels were significantly correlated with HOMA-IR in T2DM patients and healthy controls. The area under the receiver operating characteristic (ROC) curve (AUC) of OA for the diagnosis of IR was 0.689. When the cut-off value identified by Youden index was 235.8 μmol/L, the sensitivity and specificity were 70.4% and 63%, respectively. When OA combined with fasting blood glucose (FBG) to diagnose IR, the AUC increased to 0.806, which was significantly higher than that of OA (P＜0.05). @*Conclusion@#A scientific and efficient HPLC-MS/MS method for the quantitative detection of serum OA is established successfully, which provides a reliable method for the dynamic monitoring of the changes of OA levels in the patients with metabolic diseases.
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Objective@#This study was designed to evaluate the clinical value of seven combinedtumor-associated autoantibodies (7-TAAB) in the diagnosis of non-small cell lung cancer (NSCLC).@*Methods@#This is a cross-sectional study. The 81 newly diagnosed patients with NSCLC were enrolled. 46 patients with benign pulmonary diseases (BLD) and 55 healthy subjects were selected as the BLD group and the healthy control (HC) group, respectively. ELISA was used to detect the concentration of seven TAABs of p53, PGP9.5, SOX2, GAGE7, GBU4-5, MAGE A1 and CAGE in the serum of the NSCLC and the other two groups. The levels of lung cancer tumor markers CEA, NSE, SCC and CYFRA21-1 in serum were also detected in all enrolled subjects. Kruskal-wallis test was used for comparison among the three groups, Mann-Whitney test was used to evaluate the differences between the two groups, and positivity rates were analyzed by using standard χ2 tests and Fisher exact tests. The receiver operating characteristic (ROC) analyses were performed to evaluate the diagnostic efficacy of 7-TAAB or combination of 7-TAAB and traditional tumor markers.@*Results@#The serological levels of six TAABs (p53, SOX2, GAGE7, GBU4-5, MAGE A1, and CAGE) in the NSCLC group were higher than that in the BLD group (p53: Z=-4.370, P=0.000; SOX2: Z=-4.412, P=0.000; GAGE7: Z=-4.250, P=0.001; GBU4-5: Z=-2.678, P=0.025; MAGE A1: Z=-4.504, P=0.002; CAGE: Z=-4.646, P=0.001) and the HC group (p53: Z=-3.543, P=0.000; SOX2: Z=-3.383, P=0.002; GAGE7: Z=-4.893, P=0.001; GBU4-5: Z=-3.381, P=0.025; MAGE A1: Z=-3.369, P=0.001; CAGE: Z=-2.981, P=0.002),respectively. The differences were statistically significant. The comparison of PGP9.5 in NSCLC group with that in the BLD group was statistically significant (Z=-2.871, P=0.044), with that in the HC group was no difference (Z=-2.280, P=0.05). None of the seven TAABs showed a significant difference between the BLD group and the HC group (p53: Z=-1.917, P=0.917; PGP9.5: Z=-1.228, P=0.966; SOX2: Z=-1.789, P=0.325; GAGE7: Z=-0.563, P=1.000; GBU4-5: Z=-0.315, P=0.985; MAGE A1: Z=-2.310, P=0.857; CAGE: Z=-2.822, P=0.703). According to the criteria of cut-off value, the detection value of individual TAAB was judged as negative or positive. The specificity of every single TAAB to NSCLC was ≥89%, but the sensitivity was ≤39.5%. Positive in any of the single TAAB was considered as a positive result of 7-TAAB, the positive rate of 7-TAAB in NSCLC subgroups with early stages (stageⅠand stageⅡ) was considered higher than that of traditional biomarkers (7-TAAB:52.94%, CEA: 23.53%, NSE: 8.82%, CYFRA21-1∶20.59%, SCC:14.71%), and 7-TAAB was more sensitive to NSCLC patients with poor prognosis, such as advanced stages (stageⅢand stageⅣ) and moderately-poorly differentiation. The AUC of 7-TAAB was 0.734, with a sensitivity of 66.67% and a specificity of 80.20%. In coordination with 7-TAAB, CEA, NSE, CYFRA21-1 and SCC, the AUC was 0.917, with a sensitivity of 87.70% and a specificity of 81.20%.@*Conclusions@#7-TAAB, regarded as a panel of serological markers, is helpful in NSCLC diagnosis and shows broad application prospect. The detection rate of 7-TAAB in patients with early NSCLC is superior to that of traditional serum tumor markers, and the combination of 7-TAAB with CEA, NSE CYFRA21-1 and SCC could improve the diagnosis sensitivity of patients with NSCLC.
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Objective This study was designed to evaluate the clinical value of seven combinedtumor-associated autoantibodies (7-TAAB) in the diagnosis of non-small cell lung cancer (NSCLC). Methods This is a cross-sectional study. The 81 newly diagnosed patients with NSCLC were enrolled.46 patients with benign pulmonary diseases (BLD) and 55 healthy subjects were selected as the BLD group and the healthy control (HC) group, respectively. ELISA was used to detect the concentration of seven TAABs of p53, PGP9.5, SOX2, GAGE7, GBU4-5, MAGE A1 and CAGE in the serum of the NSCLC and the other two groups. The levels of lung cancer tumor markers CEA, NSE, SCC and CYFRA21-1 in serum were also detected in all enrolled subjects. Kruskal-wallis test was used for comparison among the three groups, Mann-Whitney test was used to evaluate the differences between the two groups, and positivity rates were analyzed by using standard χ2 tests and Fisher exact tests. The receiver operating characteristic (ROC) analyses were performed to evaluate the diagnostic efficacy of 7-TAAB or combination of 7-TAAB and traditional tumor markers. Results The serological levels of six TAABs (p53, SOX2, GAGE7, GBU4-5, MAGE A1, and CAGE) in the NSCLC group were higher than that in the BLD group (p53: Z=-4.370, P=0.000;SOX2:Z=-4.412, P=0.000;GAGE7:Z=-4.250, P=0.001;GBU4-5:Z=-2.678, P=0.025;MAGE A1:Z=-4.504, P=0.002;CAGE:Z=-4.646, P=0.001) and the HC group (p53:Z=-3.543, P=0.000;SOX2:Z=-3.383, P=0.002;GAGE7:Z=-4.893, P=0.001;GBU4-5:Z=-3.381, P=0.025;MAGE A1:Z=-3.369, P=0.001;CAGE:Z=-2.981, P=0.002),respectively. The differences were statistically significant. The comparison of PGP9.5 in NSCLC group with that in the BLD group was statistically significant (Z=-2.871, P=0.044), with that in the HC group was no difference (Z=-2.280, P=0.05). None of the seven TAABs showed a significant difference between the BLD group and the HC group (p53: Z=-1.917, P=0.917; PGP9.5: Z=-1.228, P=0.966;SOX2:Z=-1.789, P=0.325;GAGE7:Z=-0.563, P=1.000;GBU4-5:Z=-0.315, P=0.985;MAGE A1:Z=-2.310, P=0.857;CAGE:Z=-2.822, P=0.703). According to the criteria of cut-off value, the detection value of individual TAAB was judged as negative or positive. The specificity of every single TAAB to NSCLC was≥89％, but the sensitivity was≤39.5％. Positive in any of the single TAAB was considered as a positive result of 7-TAAB, the positive rate of 7-TAAB in NSCLC subgroups with early stages (stageⅠand stageⅡ) was considered higher than that of traditional biomarkers (7-TAAB:52.94％, CEA:23.53％, NSE:8.82％, CYFRA21-1:20.59％, SCC:14.71％), and 7-TAAB was more sensitive to NSCLC patients with poor prognosis, such as advanced stages (stageⅢand stageⅣ) and moderately-poorly differentiation. The AUC of 7-TAAB was 0.734, with a sensitivity of 66.67％and a specificity of 80.20％. In coordination with 7-TAAB, CEA, NSE, CYFRA21-1 and SCC, the AUC was 0.917, with a sensitivity of 87.70％ and a specificity of 81.20％.Conclusions 7-TAAB, regarded as a panel of serological markers, is helpful in NSCLC diagnosis and shows broad application prospect. The detection rate of 7-TAAB in patients with early NSCLC is superior to that of traditional serum tumor markers, and the combination of 7-TAAB with CEA, NSE CYFRA21-1 and SCC could improve the diagnosis sensitivity of patients with NSCLC.
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Objective To investigate the correlation between atlE gene and biofilm formation of Staphylococcus epidermidis . Methods 64 strains of clinically isolated Staphylococcus epidermidis in our hospital from June 2015 to June 2016 were collected . The biofilm formation test was used to detect bacterial biofilm .PCR was use to amplify atlE gene .Then the correlation between the atlE gene with biofilm formation was analyzed .Results 24 strains of biofilm positive bacterium were detected ,the detection rate was 37 .5% ;31 strains of atlE gene was detected ,the detection rate was 48 .4% ;atlE gene was significantly correlated to biofilm formation(P＜0 .05) .Conclusion Staphylococcus epidermidis has the ability to form biofilm ;atlE gene has a relation with biofilm formation of Staphylococcus epidermidis .
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While several lines of evidence prove that elevated concentrations of low-density lipoproteins (LDL) usually contribute to the development of atherosclerosis and its clinical consequences, high-density lipoproteins (HDL) are widely believed to exert atheroprotective effects. Hence, HDL cholesterol (HDL-C) is in general still considered as good cholesterol. Recent researches, however, suggest that this might not always be the case and that a fundamental reassessment of the clinical significance of HDL-C is warranted. The main function of HDL is to transfer the cholesterol outside the liver into the liver for catabolism.The liver′s cholesterol metabolism and other biological effects are dependent on the number of HDL particles and its proteins and lipid contents. These functions are difficult to be described simply with the HDL-C concentration. If the components of HDL particles change, they may have adverse effects on the blood vessels. Thus, high concentrations of HDL-C in plasma are not always protective factors, and some clinical trials improving HDL-C concentrations have failed to confirm a protective effect. To explore the complex relationship and pathological mechanism between HDL and atherosclerotic diseases, it is instructive for clinical application of the HDL measurement.
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Objective To investigate the clinical application value of serum lipoprotein associated phospholipase A2(Lp‐PLA2) in coronary atherosclerotic heart diseases(CAD) .Methods Using the case‐control study ,790 patients with coronary computed tomography angiography (CTA) in our hospital from October 2013 to June 2015 were selected and divided into the CAD group (352 cases) and control group (438 cases) according to the results of coronary artery CTA .According to the number of coronary artery lesion vessels the CAD group was re‐divided into three subgroups :single branch coronary artery lesion (118 cases) ,double branch coronary arterial lesions(n=107) and multiple branch coronary arterial lesions(132 cases) .The levels of Lp‐PLA2 ,hs‐CRP , TG ,TC ,HDL‐C ,LDL‐C ,glucose ,HbA1c and other indexes were measured and comprehensively analyzed .The t test or variance a‐nalysis was used to compare the means between or among groups .The correlation of different indicators was analyzed with the Pearson linear correlation analysis .Results Compared with the control group ,the CAD group was significantly higher than the con‐trols in the levels of Lp‐PLA2 ,hs‐CRP ,age ,GLU ,HbA1c and ApoB ,the differences were statistically significant(P0 .05) .Conclusion Serum Lp‐PLA2 level increase is a risk factor of CAD and could be used to assess coronary arterial atherosclerosis and number of coronary arterial lesions .
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Objective To investigate the formation of biofilm in clinical isolates of Staphylococcus epidermidis ,and to analyse the correlation between biofilm formation and antibacterial resistance of Staphylococcus epidermidis .Methods A total of 62 strains of Staphylococcus epidermidis isolated from blood specimens of inpatients with bloodstream infection ,from January 2014 to February 2015 ,were collected .The biofilm formation of Staphylococcus epidermidis was detected by using the semi‐quantitative adherence as‐say and polymerase chain reaction(PCR) amplification experiment .The antibacterial susceptibility test was carried out according to K‐B method .Results The positive rate of biofilm formation detected by using the semi‐quantitative adherence assay and PCR for icaA gene were 37 .1% (23 strains) and 43 .5% (27 strains) respectively ,and there was no statistically significant difference(P>0 .05) .There were 14 positive strains detected by both methods .The resistance rates of strains producing biofilm to antibacterial a‐gents were generally higher than those of non‐producing biofilm strains ,and there were statistically significant differences in resist‐ance rates of strains to gentamicin ,penicillin ,oxacillin ,levofloxacin and cefoxitin(P<0 .05) .All bacteria were sensitive to vancomy‐cin ,linezolid and quinupristin/dalfopristin .Conclusion There is no significant difference between the two methods in detecing bio‐film formation .The resistance rates of strains producing biofilm to antibacterial agents were generally higher than those of non‐pro‐ducing biofilm strains .
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Objective To investigate the clinical value of serum lipoprotein ( a ) concentration in evaluation of plaques characteristics for patients with coronary atherosclerotic heart diseases ( CAHD ) . Methods Using case-control method, Patients with suspicious CAHD, received coronary computed tomography angiography in the Shanghai East Hospital during October 2013 to June 2015 were enrolled.According to the results of coronary artery CTA, the patients were divided into two groups : the CAHD group (352 cases) and control group(438 cases) , the particle concentrations and mass concentration of lipoprotein(a), triglyceride, total cholesterol, HDL-C, LDL-C, glucose, HBA1c and hs-CRP and other tests were measured, the patients of CAHD group were divided into three subgroups by characteristics of coronary artery plaques including soft plaque (176 cases), calcified plaque (90 cases) and mixed plaque (86 cases), analysis were made with all these data.Using T test or variance analysis to compare the means between or among groups, the risk for CAHD was analyzed by logistic regression, the relationship between LP (a) -P and LP( a) -M were explored by linearly egression analysis, Conformance test were analyzed using kappa test.Results Compared with control group, the mean results of the CAHD group are significantly higher than that of control group, including LP (a) -P 18.5(8.3 -43.0))nmol/L vs.13.6 (7.6-32.4)nmol/L( t =-2.110), LP(a)-M 183(71 -361)mg/L vs.126(67 -293)mg/L(t =-2.063), age (62 ±9)years vs.(52 ±9)years(t=-7.691), hs-CRP 0.86(0.44-1.97) )mg/L vs.0.70(0.38-1.64)mg/L(t=-2.236), glucose (6.1 ±2.29 )mmol/L vs.(5.36 ±1.32 )mmol/L(t=-4.914), BA1c (6.13 ±0.98) % vs.(5.81 ±0.58) %(t=-4.842), APO(B) (1.09 ±0.33) g/L vs.(1.03 ±0.29) g/L( t=-2.407), all of the P values <0.05;The relative risk(RR)of age, glucose, LP( a)-P and LP ( a)-M are 1.067, 2.377, 1.384 and 1.342 respectively; Among the three types of plaques groups,the mean differences of age, TC, HDL-C, LDL-C and LP ( a)-P are statistically significant ( F=6.276,3.060,3.127,4.723,2.878;all of the P<0.05);The median of LP ( a)-P in the soft plaque group 20.3(8.3-48.2)nmol/L is higher than that of the mixed plaque group 15.7(7.3-26.0)nmol/L(P<0.05 ) and calcified plaque group 15.6 ( 8.1 -23.1 ) nmol/L ( P <0.05 ).The linearly regression equation of LP ( a) -M and LP( a)-P is Y=6.646X, r=0.939; Consistency test indicate the two methods are not consistent when used for grouping ( Kappa value is 0.557 ).Conclusions Serum concentration of lipoprotein(a) is an independent risk factor of CAHD, and the particle concentration of LP(a) is closely related to the characteristics of the plaques, especially to the soft plaque.
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Objective To investigate the influence of classically activated macrophage (M1) on the proliferation of rheumatoid arthritis (RA) fibroblast-like synovial (FLS) and osteoarthritis (OA) FLS proliferation.Methods Human monocytes leukemia cells (THP)-1 were induced into M1 by lipopolysaccharides (LPS) and interferon gamma (IFN-γ),M1 specific surface molecular markers human leukocyte antigen (HLA)-DR and CD197 were detected by flow cytometry (FCM).RA-FLS and OA-FLS were co-cultured with M1 by transwell chambers,the proliferation of RA-FLS and OA-FLS were observed by crystal violet staining assay.MTS was used to detect cytokines secreted from M1 on the multiplication of RA-FLS and OA-FLS.TNF-α and IL-12 were detected by enzyme linked immunosorbent assay (ELISA).Paried student t test was used for statistical analysis.Results THP-1 were induced into M1 by LPS and IFN-γ,the expression rates of M1 surface specific molecular markers HLA-DR and CD197 were 78.25％ and 87.96％.Crystal violet staining showed that RA-FLS and OA-FLS proliferation were significantly inhibited after co-cultured with M1 48 h,RA-FLS and OA-FLS of each vision under microscope in co-culture groups were (64 ±30) and (85 ±23) respectively,while the RA-FLS and OA-FLS in separate culture groups were (467±87) and (263±78) respectively,the difference was statistically significant (t=7.459,3.791;P＜0.05).MTS assay indirectly reflected that the cytokines from M1 suppressed RA-FLS and OA-FLS proliferation (t=-7.155,-8.111;P＜0.05).The concentration of TNF-α in cell culture supernatants secreted from RA-FLS group and RA-FLS/M1 co-culture group respectively were (0.024±0.01 1) ng/ml and (0.832±0.241) ng/ml respectively,the concentration of IL-12 from the two groups were (0.033±0.015) ng/ml and (0.372±0.122) ng/ml respectively.TNF-α from OA-FLS and OA-FLS/M1 co-culture group respectively were (0.031±0.017) ng/ml and (0.852±0.323) ng/ml,IL-12 were (0.012±0.009) ng/ml,(0.373±0.144) ng/ml.Compared with FLS separate culture group,the concentration of TNF-α and IL-12 were obviously elevated (t=-4.997,-4.777,-4.407,-4.334;P were all ＜0.05).Conclusion M1 can significantly inhibite RA-FLS and OA-FLS proliferation,this may be related to the increased concentration of TNF-α and IL-1 β in from cell culture supernatant.
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The complement cascade, as a part of innate immune system, plays a major role in phagocytosis, clearance of apoptotic cells, immune response and inflammation.As an initiator of the classical pathway, C1q not only facilitates apoptotic debris removal but also gets involved in the maintenance of vascular endothelial integrity.As a result, deficiency, excessive consumption or dysfunction of C1q leads to the imbalance of such mechanisms and increases the susceptibility of nephropathy, atherosclerosis and central nervous system diseases.Recenlty, C1q was identified as a new biomarker of aging.C1q could be a useful indicator for early diagnosis, therapy and prognosis.
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Objective To explore the serum level of Glucagon like peptide-1 in late-onset Alzheimer′s patients and its clinical significance.Methods Case control study.Collecting cerebral vascular disease fifty-five cases, diagnosed with late-onset Alzheimer′s disease sixty-one cases, type 2 diabetes mellitus fifty-one cases , type 2 diabetic patients combined with late-onset Alzheimer′s disease thirty-seven patients from the Shanghai East Hospital and partly Pudong area elderdly hospital during October 2013 to March 2014, and forty healthy persons as normal control from physical examination center of Shanghai East Hospital during September 2013 to February 2014.Measuring the concentrations of GLP-1,β-amyloid, Tau protein and other routinely used clinical tests in the serum of patients from the normal controls , cerebrovascular disease , late-onset Alzheimer′s disease, type 2 diabetes and type 2 diabetes mellitus combined with late-onset Alzheimer′s disease by ELISA method developed in our laboratory.The blood samples were also collected at three fixed time including fasting time ,1 hour after taking glucose , 2 hour after taking glucose, the concentrations of GLP-1 were determined in the LOAD group , T2DM group and the T2DM combined with LOAD group and normal control group.The concentrations of serum GLP-1 among groups were compared with single factor analysis of variance , and the concentrations of serum GLP-1 between the two groups were compared using LSD-t test.Analysing the correlation between GLP-1 and other indicators with Pearson analysis.Results The fasting GLP-1 levels of LOAD group were ( 123.4 ±20.8 ) nmol/L, and they were highest between the normal control group (78.6 ±6.0) nmol/L and the cerebral blood vessel disease group(89.0 ±8.7)nmol/L (F values were 3.46 and 1.98, P0.05).Deficient secretion of GLP-1 after taking glucose 1 hour in most of the patients of T2DM combined with LOAD group (99.1 ±14.2) nmol/L, LOAD group(73.9 ±6.6 ) nmol/L and T2DM group (96.3 ±7.0 ) nmol/L could be concluded .The GLP-1 levels of T2DM combined with LOAD group after taking sugar 2 hour were (115.4 ±18.6)nmol/L ,and were higher than that of normal levels (63.3 ±6.2) nmol/L after taking sugar 2 hour(t=4.49,P0.05).Pearson correlation analysis showed that the relationship of the levels of GLP-1 with Aβ( 1-42 ) and the levels of GLP-1 after taking glucose 1 h and 2 h were positively relative, and its coefficients of correlation were 0.401,0.436,0.722.Conclusions LOAD and T2MD are similar, and they have GLP-1 secretion shortage phenomenon after taking glucose , so monitoring dynamic change of GLP-1 after taking glucose may contribute to the auxiliary diagnosis of LOAD.
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Objective To explore the association between fibrinogen gene polymorphism(rs1049636) and serum γ′ fibrinogen level and ischemic stroke (IS) .Methods 421 IS patients and 421 age‐and gender‐ matched healthy controls ,including 283 males and 138 females ,were recruited in this assay .The plasma γ′fibrinogen concentration was measured by enzyme‐linked immunosor‐bent assay (ELISA) .Fibrinogen gene polymorphism(rs1049636) were genotyped by using PCR‐LDR assay .Results γ′fibrinogen concentrations in IS patients[(159 .4 ± 97 .4)U/dL] were significantly higher than that in control group[(114 .2 ± 73 .0)U/dL] with statistically significant difference(P<0 .001) .Single nucleotide polymorphism(SNP) analysis showed that rs1049636 C allele was significantly associated withγ′fibrinogen level ,but not associated with increased risk of IS(P=0 .077) .Conclusion An associ‐ation between increasedγ′fibrinogen level and IS existed in Chinese Han population .However ,no association between rs1049636 C allele and IS risk was observed in our study .
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Objective This study was performed to investigate the effect of hypoxia on glucose-6-phosphate isomerase (G6PI) expression and cell cycle of fibroblast-like synoviocytes from synovium of rheumatoid arthritis (RA) and osteoarthritis (OA) under hypoxia or normoxia.Methods Fibroblast-like synoviocytes were cultured with either of hypoxia (3％ oxygen) or normoxia (21％ oxygen) for 24 hours.The mRNA expression of G6PI and HIF-1α was tested by PCR quantification,while the protein levels of G6PI and HIF-1α were measured by western blot.Cell cycle was performed by FACS.T-test and Mann-Whitney U were used for statistical analysis.Results The expression levels of G6PI mRNA under hypoxia in RA were higher than those of OA (2.6±0.4 vs 1.5±0.4,P＜0.05).The protein levels of G6PI in RA were higher than those of OA (P＜0.05).The expression levels of HIF-1α mRNA under hypoxia in RA were higher than those of OA (2.9±0.8vs 1.4 ±0.4,P＜0.05).The protein levels of HIF-1α in RA were higher than those of OA (P＜0.05).The G1 phase ratio of cell cycle was decreased significantly under hypoxia than those of normoxia in RA ELs (t=1 1.31,P＜0.05).The S and G2 phase ratio of cell cycle were increased.Conclusion Hypoxia upregulates G6PI and HIF-1α expression and improves proliferation in fibroblast-like synoviocytes.
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Objective To explore the detection value of peripheral blood human cartilage glycoprotein‐39 in the patients with pri‐mary Sjogren′s syndrome(pSS) .Methods 50 patients with newly diagnosed pSS in our hospital from July 2011 to July 2014 were selected as the pSS group and contemporaneous 50 individuals undergoing physical examination were selected as the normal control group .Venous blood was sampled in all subjects and the erythrocyte sedimentation rate (ESR) ,C‐reactive protein (CRP) ,human cartilage glycoprotein‐39 levels were measured and compared .The lesion number of oral gland lymphocytes and saliva flow rate were checked and compared .Results The pSS group had significantly higher peripheral blood human cartilage glycoprotein‐39 than the normal control group (t=25 .207 ,P<0 .001) .The peripheral blood human cartilage glycoprotein‐39 level in the patients with pSS was positively correlated with the lesion number of oral gland lymphocytes (r=0 .46 ,P=0 .001) ,ESR(r=0 .48 ,P=0 .001) , CRP(r=0 .70 ,P<0 .001) ,RF(r=0 .41 ,P=0 .004) and IgG (r=0 .50 ,P<0 .001) ,and negatively correlated with the saliva flow rate (r= -0 .42 ,P=0 .003) .The eripheral blood human cartilage glycoprotein‐39 level in the patients with pSS and complications was (252 .4 ± 23 .5)μg/L ,which was significantly higher than (174 .6 ± 21 .7) μg/L in the patients without complications (t=11 .678 ,P<0 .001) .Conclusion Human cartilage glycoprotein‐39 can serve as the disease activity index of pSS and its significant increase can prompt that the patient may have complications .Human cartilage glycoprotein‐39 is also an index reflecting the disease condition of pSS objectively and comprehensively and can be widely used in clinic .
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Objective To compare the difference of kidney function evaluated by using 3 different estimated glomerular filtration rate(GFR) equations in populations .Methods Retrospectively analyzed 65 856 patients who measured serum creatinine and Cysta‐tin C at the same time ,and come from the outpatients or inpatients of the hospital .The estimated GFR (eGFR) were calculated through 3 equations ,then compared the eGFR in the population and among different groups according to different kidney functions , and then grouped the people enrolled in the study again according to the eGFR calculated by using the 3 different equations and compared the differences among groups .Results Compared with the eGFR calculated by using Creatinine equation ,the correlation coefficients of the eGFRs calculated by using the other two equations were 0 .81 and 0 .90 ,respectively ,both P<0 .05 ;The differ‐ence between the means of eGFR were 6 .19 and 1 .79 mL/(min × 1 .73 m2 ) respectively with obvious significance (P<0 .01) ,in consistency analysis .There were obvious overestimation of kidney function when using Creatinine equation to calculate eGFR .Con‐clusion There is consistence and obvious difference by using the 3 CKD‐EPI′s eGFR equations .Physicians should choose suitable equations to evaluate kidney function in different populations .