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s: Objective To explore the role of microRNA-206 in peripheral blood mononuclear cells (PBMC) in the pathogenesis and development of childhood asthma. Methods Twenty-seven asthmatic children and 25 healthy controls were enrolled in the study. Peripheral blood mononuclear cells were isolated in both healthy subjects and asthmatic children in acute attack and remission stages. Total RNAs were extracted from PBMC stimulated by PMA and ionomycin, and then the RNA was reversely transcribed into cDNA. The expressions of microRNA-206 and Kruppel-like factor 4 (KLF4) and IL-17 mRNA were detected by real-time quantitative PCR (qRT-PCR) method. Results There was signiifcant difference of microRNA-206 levels among asthmatic children in attack stage and in remission stage and normal controls (F=46.58~72.81, P=0.000). Through pair-wise comparison, the microRNA-206 levels of asthmatic children in attack stage were signiifcantly lower than those in remission stage and normal control groups (P0.05). Furthermore, a negative correlation was found between the expression of miR-206 and KLF4 (r=–0.66, P<0.01) and between the expression of miR-206 and IL-17 mRNA (r=–0.81, P<0.01) in asthmatic children in attack stage. A positive correlation was also found between KLF4 and IL-17 mRNA in asthmatic children in attack stage (r=0.70, P<0.01). Conclusions The expression of miR-206 is decreased in asthmatic children, and miR-206 might be involved in the pathogenesis and development of asthma.
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Objective To explore the role of miR-206 in peripheral blood mononuclear cells (PBMCs) of systemic lupus erythematosus (SLE) patients.Methods Human PBMCs were isolated by standard densitygradient centrifugation over Ficoll-Hypaque solution in SLE and healthy controls.Total RNAs were extracted from PBMCs which were stimulated by PMA and ionomycin,thenthe RNA was transcribed reversely into cDNA.The expression of miR-206 and Kruppel-like factor 4 (KLF4) and the orphan nuclear receptor RORγt mRNA was detected by real-time quantitative polymerase chain reaction (qRT-PCR) method.T test and Spearman's correlation test were used for statistical analysis.Results The expression of miR-206 in the PBMCs of SLE patients was significantly decreased compared with that of healthy controls (0.066±0.021 vs 0.143±0.059,t=3.136,P＜0.01).The levels of KLF4 and RORγt mRNAin the PBMCs of SLE patients were increased significantly than those of healthy controls (0.637 ±0.186 vs 0.104 ± 0.028,t=6.673,P＜0.01),(0.575±0.263 vs 0.065±0.014,t=7.386,P＜0.01).Furthermore,there was a negative correlation between the expression of miR-206 and KLF4 or RORγt mRNA in SLE patients (r=-0.627,P＜ 0.01),(r=-0.853,P＜0.01).Conclusion These results indicate that the augmented expression of KLF4 mRNA may be caused by the attenuated expression of miR-206,and the high level of KLF4 mRNA evokes the proportion of Thl7 cells in SLE patients.
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Objective To explore the role of microRNA-206 (miR-206)in peripheral blood mononuclear cells (PBMC)from rheumatoid arthritis (RA)patients.Methods 27 patients with RA and 25 healthy controls were enrolled into the current study.Human peripheral blood mononuclear cells (PBMCs)were isolated by standard density-gradient centrifugation over Ficoll-Hypaque solution in rheumatoid arthritis and healthy control volunteers.Total RNAs were extracted from PBMCs which were stimulated by PMA and ionomycin,then the RNA was transcribed reversely into cDNA.The expression of mi-croRNA-206 and Kruppel-like factor 4 (KLF4)and the orphan nuclear receptor RORγt mRNA was detected by real-time quantitative PCR (qRT-PCR)method.Student’s unpaired t-test and spearman correlation were used for statistica1 analysis. Results The expression of miR-206 in the PBMCs of RA patients was significantly decreased compared with that of the healthy controls (0.056±0.019 vs 0.138±0.057,t=3.103,P<0.01),The levels of KLF4 and RORγt mRNAin the PB-MCs of RA patients were increased significantly verves those of the healthy controls(0.604±0.183 vs 0.098±0.027,t=6.651,P<0.01;0.583±0.271 vs 0.069±0.018,t=7.438,P<0.01),Furthermore,a negative correlated between the ex-pression of miR-206 and KLF4 or RORγt mRNA in RA patients (r=-0.639,P<0.01;r=-0.842,P<0.01).Conclusion These results indicated that the augmented expression of KLF4 mRNA may be caused by the attenuated expression of miR-206,and the high level of KLF4 mRNA evokes the proportion of Th17 cells in RA patients.
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Objective To detect the level of CD4+ T cell-derived leptin(LP) in peripheral blood mononuclear cells(PBMC) from the patients with Hashimoto′s thyroiditis(HT) and to investigate its role in the occurrence and development of HT .Methods The peripheral blood samples in the patients with HT and the healthy controls were collected and separated for obtaining PBMC .The CD4+ T cells magnetic beads was adopted to separate CD4+ T cells for culturing in vitro .The LP level in supernatant was detected by ELISA .The relative expression amount of the orphan nuclear receptor RORγt was detected by real-time quantitative PCR (qRT-PCR) method .Results The LP level of CD4+ T cell culture liquid in the HT patients was markedly higher than that in the healthy control group ,difference was statistically significant (P<0 .05) .Furthermore ,the LP level in CD4+ T cell culture liquid was posi-tively related with the relative expression amount of RORγt in HT patients .Conclusion The CD4+ T cell-induced leptin level is in-creased in HT patients ,which is closely related with the occurrence and development of HT .
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Objectives To explore the role of CD4+T cell-derived leptin in peripheral blood mononuclear cells (PBMC) in asthmatic children. Methods Peripheral blood mononuclear cells were isolated from peripheral blood of both healthy subjects and asthmatic children in attack and remission stages. CD4+T cells were purified from PBMCs by mag-netic beads and were cultured in vitro. Supernatants were used to detect the levels of leptin by ELISA. The expression of the orphan nuclear receptor (ROR)γt was detected by real-time quantitative PCR (qRT-PCR) method. Results There was significant difference in CD4+T cell-derived leptin levels of asthmatic children in attack stage (68.46±13.08 pg/ml), remis-sion stage (36.73±6.13 pg/ml) and normal controls (32.82±5.79 pg/ml) (P0.05). The plasma leptin of children in attack stage and remission stage, as well as in normal subjects were 16.64 ± 3.53, 14.91 ± 3.24 and 13.72 ± 5.79 ng/ml respectively with no significant differences (P>0.05). The levels of RORγt mRNA were 0.341 ± 0.175, 0.089±0.028 and 0.068±0.018 in children with asthma during attack stage, remission stage and in normal children respec-tively (P0.05). Furthermore, the result of this study showed CD4+T cell-derived leptin positively correlated to RORγt in asthmatic children in attack stage (r=0.681, P<0.01). Conclusions CD4+T cell-derived leptin is elevated in asthmatic children in attack stage and its level is closely related to the pathological process of asthma.
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Objective To evaluate reference range for fasting venous blood cells in the healthy 51 584 elderly people from Shuyang,China.Methods Totally 1000 non-old people and 51 584 elderly people were involved in this study.Fasting venous blood cells were collected from each group of subjects using standard procedures.The collected aliquots were processed according to standard operating procedures to determine participants' complete blood counts.Non-parametric methods were employed to calculate the reference intervals and 95 ％ confidence intervals for complete blood counts by Sysmex XE-2100 blood cell analyzer.Results The reference ranges of fasting venous blood cells in elderly subjects (male,female) were [(3.25-9.45) × 109/L and (3.35-9.39) × 109/L,WBC];[(3.87-5.55) × 1012/L and (3.71-5.19) × 1012/L,RBC] ; [(116.2-169.5)g/L and(107.4-153.6)g/L,Hb] ; [(37.2-52.4) ％ and(35.2-48.6) ％,HCT] ; [(86.3-104.8)fl and (85.2-103.5) fl,MCV] ; [(27.0-33.4) pgand(26.4-32.5)pg,MCH]; [(297.1-335.4)g/L and(293.3-330.5)g/L,MCHC];[[(38.4-54.2) and (38.6-52.9),RDW-SD]; [(11.3-15.4)％ and(11.4-15.3)％,RDW-CV];[(98.8-303.8) × 109/L and (109.9-334.8) × 109/L,PLT] ; [(1.10-3.42) and (1.20-3.78) ml/L,PCT];[(11.2-15.6) fl and(11.3-15.5)fl,MPV]; [[(8.89-16.7)％ and(9.48 17.1)％,PDW];[(20.3-49.1) ％ and (20.5-48.6) ％,PLCR],respectively.13 parameters of fasting venous blood samples in elderly people had statistically significant differences compared with non-old people (all P ＜0.05).Conclusions The reference range of fasting venous blood samples in elderly people are significantly different from non-old people.It is necessary to scientifically and reasonably establish the reference ranges for fasting venous blood cells in local elderly people.
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[Objective]To compare the value of the MR imaging and arthroscopy for detecting articular cartilage erosion,and to evaluate the clinical effect of MR imaging on the correct diagnosis in the early stage of articular cartilage lesion.[Method]Twenty-six patients(27 knees)with persistent knee pain who were scheduled for arthroscopy underwent MR scanning,including 3D FS-FSPGR,FSE PDWI and FSE T1WI sequences.The results of each sequence were then compared with the arthroscopic findings.[Result]Using arthroscopic results as the standard of reference,the 3D FS-FSPGR images had the higher sensitivity(96.5%)and accuracy(95.0%)than the standard MR imaging,the 3D FS-FSPGR sequence was well consistent with the result of arthroscopic,Kappa value was higher(0.776)than the other sequences(P
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[Objective]To Study the sensitive sequence of MR for all stages of articular cartilage erosion and to evaluate the effect of MR on the correct diagnosis in the early stage of articular cartilage.[Method]One human fresh amputated knee joint and 4 normal pig knee joints were used to study and select the best sequence by T2WI,PDWI,GE,STIR,3D FS-FSPGR sequences and 6 pig oatcoarthritis(OA)model knee joints were studied by the selected best sequence,MRI was compared with correspont histological evidence to evaluate the effect of MR on the correct diagnosis of OA cartilage.[Result]The best results of tissue resolution and diagnosis rate were seen in 3D FS-FSPGR of all five sequences respectively 67.9% and 93.7%,of proteogly(PG)and collagen fiber(CF)mainly distributed in the deep cantilage.Dyeing of PG in diffent cantilage erosion was decreased while CF increasing.[Conclusion]3D FS-FSPGR is a favorable scan sequence to examine the OA changes of articular cartilage.MRI could show the pathological changes including early stage and have good veracity to the early pathological changes of OA cartilage.