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Objective@#To investigate the levels of urinary exosomal miR-194-5p in children with idiopathic nephrotic syndrome (INS) and its clinical value as a non-invasive auxiliary diagnostic marker. @*Methods@#Urine samples were collected from 101 INS children and 98 sex- and age-matched healthy children, and the levels of urinary exosomal miR-194-5p were determined by quantitative real-time polymerase chain reaction (qRT-PCR). The clinical value of urinary exosomal miR-194-5p in the diagnosis of children′s INS was evaluated by the ROC curve and correlation analysis. @*Results@#The levels of urinary exosomal miR-194-5p in INS children (2.420 [0.650,9.515] fmol/L) were significantly higher than that in healthy controls (0.360 [0.220,0.653] fmol/L, U=1 552, P＜0.01). Compared with the INS children before treatment, the levels of urinary exosomal miR-194-5p in clinical remission period decreased significantly (0.320 [0.145,0.523] fmol/L vs 0.975 [0.375,4.358] fmol/L, W=708, P＜0.01). Moreover, the levels of miR-194-5p in INS children with heavy urine protein (8.430 [7.225,13.070] fmol/L) were significantly higher than that with light urine protein (2.130 [1.180,3.090] fmol/L, U=0, P＜0.01). The area under the ROC curve (AUC ROC ) of miR-194-5p was 0.843 (95%CI: 0.789-0.897) for the diagnosis of INS children. Spearman correlation analysis showed that the levels of urinary exosomal miR-194-5p in INS children were negatively correlated with serum albumin levels (r=-0.300), and were positively correlated with serum total cholesterol levels (r=0.278) and 24-hour urine protein content (r=0.296, all P＜0.01). @*Conclusion@#The levels of urinary exosomal miR-194-5p in INS children increase obviously, and are closely associated with 24-hour urine protein, serum albumin and total cholesterol levels, indicating that urinary exosomal miR-194-5p may be serve as a new non-invasive fluid biopsy molecular marker for the auxiliary diagnosis of INS in children.
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Objective@#To investigate the levels of serum piR-015151 in patients with brucellosis and evaluate its clinical value in the auxiliary diagnosis of brucellosis. @*Methods@#Serum samples and clinical data from 73 brucellosis patients diagnosed in the Affiliated Hospital of Inner Mongolia Medical University or Zhungeer Qi Center for Disease Control and Prevention and 65 age- and gender-matched healthy controls in the Eastern Theater General Hospital of PLA during 2016 and 2017 were collected. Serum piR-015151 levels were detected by quantitative real-time polymerase chain reaction (qRT-PCR). The diagnostic value of serum piR-01515 in brucellosis was evaluated by the receiver operating characteristic (ROC) curve. @*Results@#Serum piR-015151 levels (median \[P25, P75\]) in brucellosis patients (0.216 \[0.069, 0.426\]) were significantly higher than that in healthy controls (0.036 \[0.021, 0.070\], U=834.5,P＜0.01). The area under the ROC curve (AUCROC), sensitivity and specificity of serum piR-015151 in the diagnosis of brucellosis were 0.824 (95% CI: 0.751-0.897), 75.3% and 76.9%, respectively. @*Conclusion@#Serum piR-015151 levels in brucellosis patients increase significantly, which may be a potential marker in the auxiliary diagnosis of brucellosis.
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Objective To determine the expression levels of serum miR-25 in the patients with non-small cell lung cancer (NSCLC),and evaluate its clinical significance in the screening of NSCLC.Methods Serum samples from 82 untreated NSCLC patients,including 4 with TNM satge Ⅰ,10 with TNM stage Ⅱ,11 with TNM stage Ⅲ,53 with TNM stage Ⅳ and 4 with unknown stage,and 82 healthy controls with matched age and gender were collected,and the levels of miR-25 in these samples were determined by quantitative reverse transcription PCR (qRT-PCR).The diagnostic value of miR-25 in NSCLC was evaluated by the receiver operating characteristic (ROC) curve.Results Serum miR-25 levels in NSCLC patients were significantly higher than that in healthy controls (0.017 ± 0.028 vs 0.004 ±0.004,t =4.098,P ＜0.01).The area under the ROC curve (AUGROC),sensitivity and specificity of miR-25 for the diagnosis of NSCLC were 0.818 (95％ CI:0.753-0.882),70.7％ and 80.7％,respectively.In addition,the AUCROC,sensitivity and specificity of serum miR-25 for the screening of stage Ⅰ/Ⅱ NSCLC were 0.852 (95％ CI:0.728-0.976),78.6％ and 87.8％,respectively.Logistic regression analysis showed that miR-25 was an independent risk factor of NSCLC (OR =10.84,95％ CI:5.07-23.19,P ＜ 0.01).Conclusion Serum miR-25 levels in NSCLC patients increase significantly,indicating that it may be a novel molecular biomarker for the screening of NSCLC.
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Objective To develop a method for the determination of five furostanol saponins(timosaponin N,timosaponin L, timosaponin BⅡ,25R-timosaponin BⅡ,and 25S-officinalisnin-Ⅰ)in rhizome and fibrous root of Anemarrhena asphodeloides Bge. by HPLC with the charged aerosol detector(CAD). Methods The analysis was performed on TechMate C18-ST-II(250 mm×4.6 mm,5μm)with acetonitrile:water(22:78,V/V),the flow rate of 1.0 ml/min and column temperature at 30℃. The Corona parameters were as follows:sampling rate 10 Hz,filter 5 s,and the nebulizer temperature 55℃. Results The approach showed good linearity for five saponins. The correlation coefficients(r2)for calibration curves varied from 0.9992 to 0.9998. The limits of detection(LOD)were 0.28,0.92,0.92,0.92 and 0.92 ng for five steroidal saponins,respectively. The limits of quantitation(LOQ)were found to be 0.92, 2.77,2.77,2.77 and 2.76 ng,respectively. RSD calculated from peak area of precision,repeatability and stability in 48 h were all less than 3.0%. The average recoveries of timosaponin N,timosaponin L,timosaponin BⅡ,25R-timosaponinBⅡ,and 25S-officinalis-nin-Ⅰwere 98.17%,101.37%,98.53%,97.63%,and 98.17%,respectively. Conclusion The developed method is accurate,reli-able,which could be applied to the quality control of multiple components in A. asphodeloides Bge.
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Objective To investigate the expression level of serum miR-193a-3p,miR-337-5p and miR-483-5p in esophageal squamous cell carcinoma (ESCC) patients and to explore their value for diagnosis and prognosis of ESCC.Methods Serum samples were collected from 63 ESCC patients before and after surgery in Nanjing General Hospital and Xuzhou Cancer Hospital between June 2013 and May 2014 and serum sanples of 63 age-and sex-matched healthy individuals acted as the normal controls.TaqMan Low Density Assay was used to detect the deregulated miRNA in ESCC patients and then quantitative real-time PCR was used to validate the upregulated miRNA miR-193a-3p,miR-337-5p and previously reported miR483-5p that was upregulated in oral squamous cell carcinoma (OSCC).Finally,the three miRNA were evaluated for their clinical value in the diagnosis and predicting prognosis of ESCC.Results Compared with normal controls,serum levels of miR193a-3p,miR-337-5p and miR-483-5p in ESCC patients were significantly up-regulated (0.459±0.339 vs 0.195±0.084,U =591;5.686±5.211 vs 2.476±0.808,U=605;32.545 ± 22.479 vs 19.509±10.601,U=1 037,respectively,all P＜ 0.0001) and their levels were significantly reduced after the surgical treatment (P＜0.05).The areas under the ROC curve of serum miR-193a-3p,miR-337-5p,miR-483-5p and miR-Panel were all larger than that of CEA.Univariate Kaplan-Meier analysis revealed that ESCC patients with low expression level of miR-483-5p in postoperative serum exhibited higher survival rate than those with high level(P=0.022).Conclusion Serum miR-193a-3p,miR-337-5p and miR-483-5p can be potential molecular biomarkers in the diagnosis and predicting prognosis of ESCC.
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Objective To measure and compare the expression profdes of plasma miRNA between Tibetan and Han nationality.Methods Plasma samples were taken from 246 healthy Tibetans and 128 Han individuals.The randomly selected 50 Tibetan and Han plasma samples were pooled respectively and the levels of 754 miRNAs were examined using a TaqMan Low Density Array.Two markedly differentially expressed miRNAs,miR-130a-3p and miR-629-5p,were verified in all the plasma samples by individual qRT-PCR.Results The Low Density Array results showed that the correlation coefficient of expression profiles of plasma miRNA for the Tibetan and Han population was 0.592.Compared with Han population,the expression levels of 139 miRNAs were distinctly different,in Tibetan (62 up-regulated and 77 down-regulated).The levels of miR-130a-3p and miR-629-5p were further verified to be significantly higher in the plasma from Tibetans than those in the plasma from Han population [(467 ± 27.30) × 10-5 vs (236 ± 9.69) × 10-5,p ＜ 0.01;(14.67 ±0.94) × 10-5 vs(7.58 ± 0.52) × 10-5,P ＜ 0.01] by qRT-PCR assay.Conclusion There may be marked difference of plasma miRNA expression profile between Tibetan and Han nationality.The influence of the nationality factors on miRNA profiles should be taken into account in the application of miRNAs in clinical detection in the future.
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Objective To investigate the expression levels of miRNA in kidney biopsies of child patients with nephrotic syndrome and the possibility of miRNA as potential markers in differentiating the pathologic subtypes of nephrosis.Methods Kidney biopsy specimens from 41 child patients with nephrotic syndrome,including 22 with nesangial proliferative glomeplonephritis (MsPGN),8 with minimal change disease (MCD) and 11 with endocapillary proliferative glomerulonephritis (ECPGN),were collected,and adult nephridial tissues from 8 patients without renal inadequacy were selected as controls.The expression levels of miR-191,miR-151-3p,miR-150,miR-30a-5p and miR-19b in nephridial tissues were detected by RT-qPCR,and their correlations with renal function related parameters were analyzed.Results Compared with the controls,the miR-191 levels in kidney tissues of child patients with nephrotic syndrome increased significantly (P ＜ 0.01),while the miR-151-3p levels decreased obviously (P ＜ 0.01).The expression levels of miR-150 in MCD patients were significantly lower than those in MsPGN and ECPGN patients and the controls (P ＜ 0.05).The expression levels of these miRNAs were positively correlated with serum IgG,TP and Cr levels,but negatively with serum TC levels (P ＜0.05).Conclusion The expression levels of miRNA in kidney tissues of child patients with nephrotic syndrome are related to pathological typing of nephrosis,and miR-150 may be a potential marker which may differentiate MCD from other subtypes of nephrosis.
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Objective To determine the serum levels of miR-16,miR-126 and miR-221 in type 2 diabetes (T2DM)patients with or without microvascular complications,and further evaluate their clinical significance.Methods The serum levels of miR-16,miR-126 and miR-221were examined in 55 T2DM patients,55 T2DM patients with microvascular complications and 5 5 healthy controls using quantitative real-time PCR (qRT-PCR).The levels of fasting blood glucose,triglycerides,choles-terol,high density lipoprotein,low density lipoprotein and others biochemical parameters were determined by biochemical an-alyzer,and the diagnostic usefulness of the three miRNAs for T2DM patients and patients with microvascular complications were assessed by ROC curve analysis and logistic regression analysis.Results Compared with healthy controls [miR-1 6 (14.35±1.00)×10-5 ,miR-126(11.75±1.47)×10-5 and miR-221(32.26±3.98)×10-5 ],the miR-16,miR-126 and miR-221 expression were significantly increased in T2DM patients [miR-16(23.74±2.70)×10-5,miR-126(25.01±4.13)× 10-5 and miR-221(84.76±11.79)×10-5 ]and T2DM patients with microvascular complications [miR-16(43.74±9.61)× 10-5 ,miR-126(17.66±2.20)×10-5 and miR-221(82.52±12.48)×10-5 ].The area under ROC curve (AUCROC)of miR-16,miR-126 and miR-221 for T2DM patients were 0.63 (95%CI 0.53~0.74),0.64 (95%CI 0.54~0.74)and 0.74 (95%CI 0.65~0.83),respectively.For T2DM patients with microvascular complications,the area under ROC curve were 0.75 (95%CI 0.66~0.84),0.62 (95%CI 0.52~0.73)and 0.73 (95%CI 0.64~0.83),respectively.Furthermore,logistic re-gression revealed that the three miRNAs were novel independent risk factors for T2DM and T2DMC.Conclusion The levels of miR-16,miR-126 and miR-221 were significantly increased in the serum of T2DM patients with or without microvascular complications,and can be used as potential non-invasive biomarkers and risk factors for T2DM patients and T2DM patients with microvascular complications.
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Objective To determine the levels of miR-193a-3p in serum from patients with renal clear cell carcinoma,and eval-uate the potential of miR-193a-3p asa molecular biomarker of renal clear cell carcinoma.Methods Serum samples were taken from 107 untreated hospitalized patients with renal clear cell carcinoma (TNM Ⅰ grade 76 cases,Ⅱ grade 16 cases,Ⅲ grade 2 cases,Ⅳ grade 8 cases and unknown 5 cases)in Nanjing General Hospital from 2010 to 2014 year and 107 age-and gender-matched healthy individuals to determine the level of miR-193a-3p by quantitative reverse-transcription PCR (qRT-PCR). The early diagnostic value of miR-193a-3p for renal clear cell carcinoma was assessed by ROC curve.Results qRT-PCR confirmed that serum miR-193a-3p expression levels was significantly elevated in cancer than in normal controls (3.294± 1.526 vs 1.944±0.600,P =0.000).TNM Ⅰ grade (3.411±1.676 vs 1.944±0.600,P =0.000),Ⅱ grade (2.926±0.927 vs 1.944±0.600,P =0.001),Ⅳ grade(2.926±0.894 vs 1.944±0.600,P =0.000)is significantly higher than that in con-trol group.The area under the ROC curve (AUC)of miR-193a-3p was 0.820 (95% CI,0.756 ~ 0.883),demonstratinga high sensitivity (80.3%)and specificity (93.5%)for the early diagnosis of renal clear cell carcinoma.Conclusion MiR-193a-3p content was significantly elevated in serum from renal clear cell carcinoma and has a high accuracy in diagnosing ear-ly cancer,which holds potential as molecular biomarker for renal clear cell carcinoma.
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College students' self cognition of the learning effectiveness influences their learning behavior greatly. Focusing on college students' self cognition of their learning effectiveness, and then standardizing their learning behavior is one of the important work of educators in colleges. Therefore this paper has designed the questionnaire of the influencing factors of learning effectiveness self cog-nition, used SIMPP analysis method to analyze it, and set up the relation model. The analysis shows that college students' self cognition of learning effectiveness is influenced not only by students' dili-gence level and the recognition degree of specialized subject, but also by the factors such as the attitude when you get the bad results. The result provides data basis and scientific basis for future practice and relevant research for educators in colleges.
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In recent years, lower and lower class attendance has plagued the quality of uni-versity teaching. To solve this problem, this study conducted a questionnaire survey in accordance with the concepts of the Education Reform microscopic systems engineering and the method of SIMPP. The results showed that factors affecting student classroom attendance included two aspects: the sub-jective and objective factors. Indicators related to the subjective factors were: the personal attitude when faced with failure exams, the personal learning interest and personal grasp of the main source of knowledge. Indicators related to the objective factors were: school and teachers. Also, this study gave some suggestions on how to improve students' classroom attendance to provide data basis and refer-ence for further study on class attendance.
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MicroRNAs ( miRNAs) are a kinds of small non-coding RNA that regulate the expression of gene at the posttran-scriptional level and play important roles in a variety of physiological and pathological processes such as proliferation , differentiation and apoptosis.Recently, it has been reported that miRNAs expression abnormalities are closely associated with the initiation and devel -opment of glioma .Moreover , the expression levels of some miRNA are specifically involved in the molecular pathogenesis of glioma , revealing their great potential as a class of novel biomarkers for cancers and therapeutic targets .In this review , we summarizes the lat-est researches on miRNA and glioma .
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Objective To evaluate the diagnostic and therapeutic significance of serum free light chain (sFLC) in primary systemic(AL) amyloidosis. Methods Twenty-five patients with AL amyloidosis,including 18 men and 7 women with a mean age of 54(47-77) years old, were enrolled from October, 2005to May, 2010. sFLC was measured by immunoturbidimetric assay. The type of monoclonal light chain was judged upon sFLC κ/λ and its sensibility was compared with serum immunofixation and immunohistochemical analysis. Four patients were treated with M (T)D (melphalan/thalidomideand, dexamethasone), one with VD (velcade and dexamethasone) and four with high-dose melphalan followed by autologous stem cell support. The changes of sFLC were serially determined before and after treatment. Results Among the 25 patients with AL amyloidosis, two were κ light chains of precursor protein and 23 were λ light chains. Mean plasma cell in bone marrow was 3.5% (0-15%). Nineteen (76%) patients had abnormal elevated sFLC and abnormal κ/λ ratios, and 17(68% ) patients with immunofixation positive. The sFLC test had similar sensitivity as serum immunofixation (P = 0. 727 ). Twenty-one (84%) patients were shown to have either κor λ immunoreactive amyloid deposits on biopsied tissues. The sFLC test combined with serum immunofixation allowed the M protein to be detected in 22 (88%) patients. The positive rates of immunohistochemical analysis combined with sFLC test and/or serum immunofixation were 96%. Four patients with hematologic response showed obvious improvement in visceral organ involvement, but illness of 5 patients without hematologic response kept stable or progressed. Conclusions sFLC test is a sensitive qualitative and quantitative method to detect M protein. Preliminary data show the patients with obvious sFLC level decrease and/or κ/λ recovery to normal may have a high percentage of improved organs function. sFLC is critical index in diagnosing AL amyloidosis, which might help efficacy assessment.
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Objective To investigate possible changes of lipoprotein(a) [Lp(a)] and oxidized Lp (a) [ox-Lp(a) ] levels after PCI and it mechanisms. Methods Bloods were selected from 75 patients with ACS undergoing PCI, and at 24 hours, 2 and 3 days, and 6 months pre-and post-PCI treatment, and from 29 control patients pre-and post-coronary angiography without undergoing PCI. The levels of Lp(a) , ox-Lp(a) , Lp(a) immune complexes (IC) and its autoantibody were determined by ELISA. The extents of CAD were determined by coronary angiography. The differences of variants pre-and post-operations were analyzed by paired samples t test. The differences of levels of Lp(a) and ox-Lp(a) among time points after PCI were analyzed by ANOVA. Correlations between Lp(a) and ox-Lp(a) , and between angiographic variables and Lp(a), ox-Lp(a) levels were calculated. Results Compared to pre-PCI, Lp(a) [233.10 (152.86-328.79) mg/L vs 202.05 (106.15-271.42) mg/L, t=6. 81, P<0.01], ox-Lp(a) [19.05 (10.98-31.80) mg/L vs 10. 51 (4.98-17.97) μg/ml, t = 13. 22,P <0. 01] and Lp(a)-IC [2.72 (1.604.91) AU vs 2. 11 (1.04-3. 97) AU, t = 3. 34, P < 0. 01 ] levels significantly increased immediately in post-PCI, while its antoantibody levels significantly decreased (A = 0. 81 ± 0. 33 vs A = 0. 72 ± 0. 28, t = 5.58, P < 0. 01). Strong correlations were noted between levels of ox-Lp( a) and Lp( a) both in pre-PCI (r =0. 66, P <0.01) and post-PCI (r = 0. 62, P <0. 01). PCI resulted in rapidrise of Lp(a) and ox-Lp(a) levels and then decreased quickly in 24 hours, returned to baseline in 2-3 days. The changes of Lp(a) and ox-Lp(a) levels in pre-and post-PCI were positively related with severity of ACS. In contrast, in the angiography-only control group, no significant changes were noted in Lp(a) , ox-Lp(a) , Lp(a)-IC and Lp(a) autoantibodies levels between the pre-and post-angiography samples. Conclusion PCI results in acute plasma acute increases of levels of Lp(a) and ox-Lp(a) ,and the changes are related with lesion severity of the coronary artery.
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Objective To explore the effects of low density lipoprotein(LDL) containing immune complexes(IC) on the mRNA expression of matrix metalloproteinase-1(MMP-1) and matrix metalloproteinase-1(TIMP-1) in U937 cells.Methods U937 cells were incubated with native LDL,oxidized LDL or LDL-IC,respectively.mRNA expression of matrix metalloproteinase-1(MMP-1) and tissue inhibitor of metalloproteinase-1(TIMP-1) in U937 cells were measured by reverse transcriptase polymerase chain reaction(PT-PCR).Results Normal U937 cells hardly expressed MMP-1,but highly expressed TIMP-1.N-LDL had no effect on the expression of MMP-1 and TIMP-1 mRNA.Ox-LDL can induce slightly expression of MMP-1 and markedly down-regulate expression of TIMP-1.Similarly,nature and oxidized LDL-IC had significantly more effect on the mRNA expressions of MMP-1 and TIMP-1 than that of control(P
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Objective To isolate and identify human anti-oxidized low density lipoprotein (LDL) autoantibodies.Methods Oxidized LDL (Ox-LDL) autoantibodies were isolated from eight healthy subjects by affinity chromatography with using an Ox-LDL cross-linked Sepharose 4B column.Results Three elution peaks of Ox-LDL autoantibodies were washed out from the Ox-LDL column by NaHCO3,acetate and KCNS buffer in turn and collected respectively.Ox-LDL autoantibodies were isolated from all the selected healthy subjects.All the three elution peaks had good reactivity with Ox-LDL.Among them,the elution peaks by KCNS buffer had high protein level and strong reactivity with Ox-LDL.The types of these antibodies included IgG and IgM, and IgG was the main type.Conclusion The isolated Ox-LDL autoantibodies had strong reactivity with Ox-LDL.
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Objectives To explore cholesteryl ester transfer protein(CETP)level in male patients with different degrees of coronary artery lesion and its clinical significance.Methods ELISA was used to measure CETP of 42 male patients with coronary heart disease(CHD)and 49 healthy male controls.The patients of CHD groups were subdivided into mono-vessel,ambi-vessel,multi-vessel lesion groups;localized,diffuse lesion groups;and mild,severe stenosis groups according to coronary angiography.Results The CETP level of patients with CHD(1.37?1.07 mg/L)was significantly higher than that of healthy control(0.99?0.53 mg/L)(P
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Cholesteryl ester transfer protein(CETP) plays an important role in the transfer and exchange of cholesteryl esters,phospholipid and triacyglycerols.It affects lipid composition,concentration,particle size and function of lipoproteins, partcipating in the development of atherosclerosis.The role of CETP in human atherosclerosis has not been fully elucidated.Gene variations of the CETP were associated with altered CETP plasma levels and activity,lipid metabolism disorder.The relationships between CETP gene variation,lipid parameters and cardiovascular risk was reviewed.
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Objective: To study the effect of Anoectochilus roxburghii on the oxidation of lipoprotein. Methods: The effects of Anoectochilus roxburghii on low density lipoprotein(LDL) oxidative modification were assessed by the formation of conjugated dienes,thiobarbituric acid-reactive substance(TBARS) and the relative electrophoretic migration. Results: The addition of Anoectochilus roxburghii(1 ?l/ml) to the reaction mixture prolonged the lag phase by 1.5 times [from(8.8?0.4)min to(13.0?0.8)min].Anoectochilus roxburghii reduced LDL oxidative susceptibility in a dose dependent manner.In addition,Anoectochilus roxburghii resulted in a significant,dose dependent,reduction of TBARS contents and the decrease in LDL electrophoretic migration as compared to the control. Conclusion: These results indicated that Anoectochilus roxburghii has effective antioxidative capacity by reducing oxidative susceptibility and peroxidation degree of LDL.
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High density lipoprotein(HDL) is an antiatherogenetic lipoprotein and low level of HDL-cholesterol(HDL-C) in plasma is a risk factor for coronary heart disease.Therefore,raising the level of HDL-C by pharmaceuticals,oestrogen replacement and biologic therapy for dyslipidemia exhibit great clinical significance.