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OBJECTIVE@#To investigate the effect of the exosomes from bone marrow mesenchymal stem cells (BMSCs) transfected with silence plasmid of Piezol small interference RNA (siRNA)on osteoarthritis (OA) animal model.@*METHODS@#Twenty male SD rats with specific pathogen free (SPF) were selected, ranging in age from 5.46 to 6.96 months, with a mean of (6.21± 0.75) months;and ranging in weight from 385.76 g to 428.66 g, with a mean of (407.21±21.45) g. BMSCs were extracted. The siRNA silencing plasmid of piezo1 was constructed by siRNA technology. After lentivirus was transfected into BMSCs, the exosomes were extracted. At the cellular level, BMSCs were divided into blank plasmid group and siRNA silencing plasmid group according to whether siRNA-Piezo1 was transfected or not. The osteogenic induction ability of siRNA-Piezo1 on BMSCs was detected by RT-PCR and Western blot. At the animal model level, the OA model was established by surgical resection of cruciate ligament of knee joint.According to different treatment schemes, SD rats were divided into 4 groups:blank control group, model group, BMSCs group and exosome group. SD rats in the blank control group were not treated. In the model group, the cruciate ligaments of rats were excised and OA animal model was established. In BMSCs group, BMSCs were injected into knee joint under CT guidance after OA model establishment, and the cell volume was 5×10@*RESULTS@#The lentivirus transfection efficiency was(92.11±4.22)%. RT-PCR showed that the relative expression of Piezo1 mRNA in blank plasmid group was 1.07±0.06, which was significantly different from that of 0.31±0.01 in siRNA silencing plasmid group (@*CONCLUSION@#Piezo1 siRNA silencing vector can promote the differentiation of BMSCs into chondrocytes and effectively inhibit the progression of OA, so as to delay the disease of OA.
Subject(s)
Animals , Male , Rats , Chondrocytes , Disease Models, Animal , Exosomes/genetics , Mesenchymal Stem Cells , Osteoarthritis/therapy , RNA, Small Interfering/genetics , Rats, Sprague-Dawley , Tomography, X-Ray ComputedABSTRACT
<p><b>OBJECTIVE</b>To explore the expression characteristics of new mechanosensitive ion channel Piezo1 protein in stress models of human degenerative chondrocytes.</p><p><b>METHODS</b>The stress stimulation model of human degenerative chondrocytes in vitro was constructed. Multi-channel cell stretch stress loading system FX-4000T was used to treat chondrocytes. According to the results of pre-test, the loading frequency of 0.5 Hz and the cell elongation of 20% were loaded. According to cell processing time, it was divided into 0 h, 2 h, 12 h, 24 h and 48 h mechanical stress group. The RT-PCR and Western-blot were used to test the expression of the Piezo1, also the Laser scanning confocal microscope (LSCM) was used to test the intensity of the fluorescence of the Piezo1.</p><p><b>RESULTS</b>(1)The result of the RT-PCR showed that the expression of the Piezo1 in the 2 h group was higher than the 0 h group(13.917, 0.037 1, <0.05). The expression of the piezo1 in the 24 h group was the highest. While the expression of the piezo1 in the 48 h group was lower than the expression of the piezo1 in the 24 h group(13.917, 0.049 5, <0.05). (2)The result of the Western-blot showed that the 2 h group was higher than the 0 h group(19.341, 0.037 1, <0.05). The expression of the 24 h had the highest expression which was higher than the 48 h group(19.341, 0.017 7, <0.05). (3)The Piezo1 protein was extensively expressed in the cytoplasm and nucleus of the nucleus pulposus cells. And with the increase of stress processing time, the fluorescence intensity of the protein also increased.</p><p><b>CONCLUSIONS</b>In human degeneration cartilage cells, the new mechanio sensitive ion channel Piezo1 protein has a trace expression. After loading periodic mechanical tensile force, the expression of Piezo1 protein increases with time dependence.</p>
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Objective To investigate the effect of arthroscopic treatment of massive rotator cuff tears. Methods 62 patients with massive rotator cuff tears underwent arthroscopic treatment from January 2010 to February 2016 were enrolled in the study and divided into elder group (≥ 65 years old, n = 23) and control group (< 65 years old, n = 39) according to age. Scores of American Shoulder and Elbow Surgeons scale (ASES), University of California Los Angeles (UCLA) and Constant-Murley were compared before surgery and at 12 months after surgery between the two groups. Results The age in elder group was significantly higher than that in control group (P < 0.05). There were no significant differences in baseline data including gender, disease duration, etiology and history of shoulder trauma between the two groups (P > 0.05). No perioperative complication occurred in all cases and all surgery were completed safely and successfully. The scores of ASES, UCLA and Constant-Murley at 12 months after surgery in the two groups were all significantly higher than that before surgery (P > 0.05). There were no significant differences in the scores of ASES, UCLA and Constant-Murley before surgery and at 12 months after surgery between the two groups (P > 0.05). Conclusions Arthroscopic minimally invasive surgery could effectively improve shoulder function in patients with massive rotator cuff tears, and it could also achieve good results in elderly patients and is worthy of clinical application.
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Objective To evaluate the therapeutic effects of aminoguanidine(AG) on cerebral ischemia-reperfusion damage in rats. Methods The intravascular thread models with 2 h of occlusion and 22 h of reperfusion were made in the rats.The brain infarction size and the degree of blood brain barrier(BBB) disruption in the ischemic regions were evaluated by staining with 2,3,5-triphenyl tetrazolium chloride and observing with Evans blue fluorescence microscope.HE staining was utilized for observing neutrophil infiltration. Results The brain infarction(volume,) the area of BBB disruption and the degree of neutrophil infiltration were dramatically decreased in the treatment group as compared to the control group(P
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Objective To study the relationship of microinflammation,nutrition and common carotid artery intima-media thickness(CCA-IMT) for the early prevention and interference of ischemic stroke. Methods(CCA-IMT was) measured by carotid arterial ultrasound in 250 elderly subjects.The levels of serum high-sensitivity C-(reactive) protein(hsCRP),ferritin,albumin,pre-albumin and transferrin were assayed at the same time.According to the results of CCA-IMT,all the subjects were divided into five groups: