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Objective To evaluate the repeatability and stability of double antibody sandwich ELISA kit for micro-quantitative soluble complement receptor 1(sCR1) in human serum and to understand its practical application effect .Methods 50 patients with middle and advanced liver cirrhosis and 50 individuals undergoing physical examination served as the liver disease group and normal control group respectively .The mouse anti-human CD35 monoclone antibody ,rabbit anti-human sCR1 polyclonal antibody and goat anti-rabbit IgG labeled by horseradish peroxidase served as the envelope antibody ,sandwich antibody and detection antiboby .The purified recombinant human sCR1 protein served as the standard substance .The human serum micro-quantitative double antibody sandwich CR1 ELISA kit was established .Then the repeatability and stability tests were performed .Then the sCR1 protein level of two group of serum was detected by this kit .Results The linear range of double antibody sandwich ELISA for detecting human se-rum micro-quantitative sCR1 protein was 15 .60 -250 .00 ng/mL ;the regression equation of sCR1 protein concentration to absor-bance value was Y=112 .10X2 +18 .21X+1 .694(r2 =0 .998);in the repeatability test ,the intra-batch relative standard deviation (RSD) in high and low concentrations of standard substance detection value was 6 .20% and 7 .40% respectively ,the inter-batch RSD was 6 .70% and 7 .90% respectively ;in the stability test ,RSD was not more than 0 .01;the serum sCR1 expression level in the liver disease group was significantly higher than that in the normal control group (P<0 .01) .Conclusion The human serum double antibody sandwich ELISA kit for detecting human sCR1 has wide linear range ,good repeatability ,is easy to be stored and suitable for clini-cal and scientific research detection work .
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Objective To observe the damage degree and expression pattern of Caveolin-3 mRNA by ischemia-reperfusion injury in rabbits of skeletal muscle cell at different phases.Methods In this study,from April 2013 to December 2013,30 lower limbs of 15 Chinese White Rabbits were used and divided into two groups:all the left lower limbs were experimental group,which were made as an experimental model of ischemia-reperfusion injury by occluding left common iliac artery using noninvasive vascular.All the right lower limbs without surgical treatment were the control group.Gastrocnemius samples were obtained at 4h and 8h after reperfusion and handled by HE staining and observed by optical microscopy.By Real-time PCR,Caveolin-3/GAPDH mRNA were detected.Results HE stain showed:in control group,there was no edema,degeneration and inflammatory cell infiltration; in experi-meatal group,muscle cell degeneration had occured at ischemic 5 h.The edema was aggravated,a large number vacuole were formed and inflammatory cell were infiltrated at 4 h reperfusion.Reperfusion injury at 8h significantly reduced compared to 4 h.The Caveolin-3/GAPDH mRNA expression levels by SPSS 19.0 showed:Control group:1.026 ± 0.065,1.004 ±0.037,1.022 ±0.051,experimental group:1.159 ±0.073,1.445 ±0.053,1.208 ±0.058 at ischemic 5 h,4 h and 8 h reperfusion,respectively.On-line analysis of variance cases of ischemic 5 h and 4 h reperfusion and 8 h reperfusion,the experimental group than the control group were increased,with statistical significance (P ＜ 0.05).The experimental group of ischemic 5 h and 8 h reperfusion was no significant difference (P ＞ 0.05).It showed Caveolin-3 mRNA expression levels in ischemia-reperfusion 8 h group returned to normal.There was significant statistical difference between the ischemic 5 h and 4 h reperfusion (P ＜ 0.05).There was significant statistical difference between the 4 h reperfusion and 8 h reperfusion (P ＜ 0.05).Conclusion The expression of Caveolin-3 in experimental group showed a trend of first increased and then decreased.The expression levels of Caveolin-3 mRNA in skeletal muscle cells after ischemia-reperfusion injury is consistent with the development and progression of muscle cell damage.The results indicate that Caveolin-3 may play a control role in the injury and recovery of skeletal muscle cell.
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Objective To explore the value of echo-tracking technique in the detecting the ipsilateral external iliac artery elasticity after gunshot wound in pig limbs.Methods Two-dimensional ultrasound imagings and elasticity parameters of external iliac artery of fifteen pig limbs were obtained respectively,and the results were compared with pre-injury groups.The animals were sacrificed after ultrasonography.Pathological examinations of adjacent external iliac artery of injured pig limbs were analyzed.Results Two-dimensional ultrasound imagings of external iliac arteries had no significant changes post-injury.The changes of elasticity parameters were significiant differences in the injured group comparing with the pre-injured group (P ＜0.05),including the increased stiffness parameter(β),pressure-strain elasticity modulus (Ep)and the decreased arterial compliance(AC).Pathological result showed that the internal elastic lamina of artery detected were flat,endothelial cells came off discontinuously and structure of them were undefined.Conclusions Echo-tracking technique can find the elastic changes of adjacent artery indirect injured by gunshot wound sensitively and which can suggest the occurrence of vascular indirect injury.
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Objective:To investigate the effect of interleukin-1β (IL-1β) on epithelial-mesenchymal transition (EMT) and cytoskeleton rearrangement of renal tubular epithelial cells.Methods:Immortalized renal tubular epithelial cell line NRK52E was cultured in vitro with IL-1β (30 μg/L) for 3 days and 6 days,then the cell morphology was observed;The mRNA expressions of α-smooth muscle actin (α-SMA),cytoskeleton components β-actin and α-tubulin were semi-quantitative examined by RT-PCR.The protein expression of α-SMA and arrangements of β-actin and α-tubulin were assessed by immunofluorescent staining.Results:After induced by IL-1β for 3 days and 6 days in vitro,the mRNA and protein expression of α-SMA increased significantly compared with corresponding control cells (P<0.001),it prompted that NRK52E cells underwent EMT;At the same time,the cell morphology also changed,from a typical multilateral paving stone to fibroblast-like appearance,with multiple processes; Cytoskeletal protein β-actin mRNA expression was also slightly increased (P<0.05).The distributions and arrangements of β-actin protein were also changed,from cell membrane transferred to peri-nucleus and cytoplasm,moreover it formed fiber bundle-like structures.However,another cytoskeleton protein α-tubulin in IL-1β induced cells,neither it's mRNA expression nor it's distribution had significant differences compared with the control group.Conclusion:IL-1β can induce NRK52E cells undergoing EMT in vitro,cell morphology changes into fibroblast-like appearance with multiple processes,and also the cytoskeleton protein β-actin expression increases and rearrangement occurrs.However,there was no changes onα-tubulin.
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Objective To investigate the clinical characteristics of postinfectious irritable bowel syndrome (PI-IBS) in Qingdao. Methods Two hundred and four PI-IBS and 2068 non-PI-IBS patients were investigated with questionnaire including general information, symptoms and quality of life scores with microecological study before and after therapy. Results (1) The morbidity rate of PI-IBS in female was 2. times of that in male, which was similar to that in non-PI-IBS. (2) Brainwork labors dominated in both PI-IBS and non-Pl-lBS patients. (3) As to the simultaneous presence of extra-gastrointestinal symptoms,there was no statistical difference between the rate of physical symptoms in PI-IBS and non-PI-IBS patients (X<'2>= 10. 5, P>0.05) ,but the rate of mental symptoms was higher in PI-IBS than in non-PI-IBS patients, and the difference was significant(X<'2>= 28.7, P<0.05). (4)The alteration of intestinal microflora rate in PI-IBS was obviously higher than that in non-PI-IBS patients. (5) The quality of life scores in PI-IBS was improved after treatment with Birid Triple Viable , and there was significant difference(t =3. 8, P<0.01),but there was no statistical difference in non-Pl-IBS (t = 1.5, P>0.05). Conclusion There was some difference in certain clinical characteristics between PI-IBS and non-PI-IBS patients in Qingdao.