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Objective:To investigate the therapeutic effect of rat bone marrow mesenchymal stem cells (BMSCs) transfected with recombinant rat platelet-derived growth factor BB (rrPDGF-BB) gene on the distraction osteogenesis.Methods:From October, 2019 to June, 2020, 48 batches of BMSCs were cultured from 48 young SD rats, 24 of which were transfected with rrPDGF-BB gene by lentivirus. Meanwhile, other 72 male adult SD rats were randomly selected to establish the right femoral distraction osteogenesis model. The rats were equally divided into 3 groups. PBS, BMSCs without intervention and BMSCs transfected with rrPDGF-BB gene were injected into the distraction space of each group of rats assigned as Blank group, Negative group and Experimental group, respectively. Results of the experiment were evaluated by means of imaging and immunohistochemistry. P<0.05 indicated a statistically significant difference. Results:The cultured BMSCs grew well. The expression of CD34(0.1%) and CD45(2.8%) in the third generation of BMSCs was low, and that of CD29 (95.1%) was high, which was consistent with the phenotype of BMSCs described in literatures. After transfection, the expression of green fluorescence gradually increased with the extension of transfection time, confirming the success of transfection. After 14 days, all rats reached the expected distance of distraction. The rats were observed at assigned time points in 2, 4 and 8 weeks. The photos of femur specimen showed that continuous callus could be seen in the experimental group, the hardness and colour were close to the normal bone tissue, and the activity of the distraction space was poor, which was lower than that of the blank group. X-ray examination showed that there were more new callus in the experimental group, and the bone marrow cavity was re-canalized earlier than that of the blank group; Micro-CT examination, in sagittal plane, showed that the distraction space of the experimental group healed well, the broken end was connected, and the recanalization of bone marrow cavity was earlier than that of the blank group; Micro-CT parameters of each group showed that trabecular thickness[(0.297±0.005) mm], trabecular number [(1.663±0.032) mm], bone volume fraction[(59.832±2.187)%] and bone mineral density[(0.586±0.014) g/cm 3] of the experimental group were the greatest, while trabecular separation[(0.399±0.051) mm] of the experimental group was the smallest. There was statistical difference between each group( P < 0.05); HE staining and VEGF immunohistochemistry showed that the vessels and chondrocytes formed earlier and were more in the experimental group than that in the blank group. In 8 weeks, the new callus joined into one piece under the microscope in the experimental group, and the bone marrow cavity was re-canalized with a large number of red blood cells. Conclusion:Studies have shown that BMSCs transfected with rrPDGF-BB gene can promote the formation of callus in the distraction area of rats, shorten the mineralisation time of new callus, and promote the maturation of new bone in the area of distraction osteogenesis.
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To investigate the effect of MMP-26 on human glioma angiogenesis and the possible mechanism.Methods The MMP-26 plasmid and empty plasmid pcDNA3.1 were stably transfected into U251 cells to establish a nude mice xenograft model,and then an in vitro human tumor tissue-based three-dimensional angiagenic model.Tissue disks were visually assessed over time to determine the percentage of wells that developed an angiogenic response(I％) and the density and length of neovessel growth were graded at intervals using a semiquantitative visual growth-rating scheme (angiogenic index,AI,0-16scale) in groups of MMP-26 transfected U251 cells (U251-MMP-26),pcDNA3.1 vector-transfected U251 cells (U251-pcDNA3.1) and non-transfected U251 cells (U251).RT-PCR and immunohistochemistry were used to detect the expression of mRNA and protein of MMP-26 and VEGF in groups of U251-MMP-26,U251-pcDNA3.1 and U251.Immunohistochemical localization of CD31 was determined in the endothelial tubes invading the fibrin-thrombin clot matrix.Results Immunohistochemical endothelial cell markers CD31 was positive in the vascular tubes invading the fibrin-thrombin clot matrix,confirming their endothelial origin.The angiogenesis results showed that difference of length of micro capillaries,density of branches,and the area occupied between U251-MMP-26 groups and control groups were significant.The percentage of tumor implants that developed invasion (I％) and the angiogenic index AI in U251-MMP-26 group on day 14 were higher than those of U251-pcDNA3.1 group and U251 group (P ＜ 0.05).The trends of I％ and AI in 14 days were significant compared with those in control groups.The expression of mRNA and protein of MMP-26and VEGF in U251-MMP-26 group was significantly higher in U251-MMP-26 group than those in U251-pcDNA3.1 group and U251 group(P ＜0.01).Conclusion The effect of MMP-26 on promoting glioma angiogenesis may be related to the increased expression of VEGF,which can be used as targets for anti-tumor therapy.
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Objective To explore the relationship between dyslipidemia and urinary stone formation.Methods The clinical data of 427 patients diagnosed with urolithiasis in our hospital during January 2015 to May 2016 were collected.Among them,272 men accounting for 63.7%,155 women account for 36.3%.The average age were 53 (43-63).218 cases were kidney stones,accounting for 35.6%;158 cases were ureteral calculi, accounting for 25.8%; 23 cases were kidney stones with ureteral calculi, accounting for 3.8%;28 cases were bladder calculi, accounting for 4.6%.At the same time,950 age and gender matched healthy controls were collected.Among them,570 men accounting for 60%, 380 women account for 40%.The average age were 53 ( 48-60 ).All of them had undergone renal ultrasound to excluded urolithiasis.The difference between lipid level and incidence of dyslipidemia in patients with urolithiasis were observed. The relationship between lipid level and serum UA, urine pH and stone composition was evaluated and analyzed with logistic regression.Results The average serum TC,TG,HDL-C levels of patients with urolithiasis were 4.34mmol/L, 1.38mmol/L, 1.25mmol/L, which levels were 4.32mmol/L,1.09mmol/L,1.40mmol/L in healthy controls.Significant difference were seen between the two groups ( P <0.05 ).The average serum LDL-C was 2.63mmol/L in patients with urolithiasis and 2.65mmol/L in healthy controls.No difference were seen between the two groups ( P=0.241).31.6% of patients with urolithiasis had different degree of dyslipidemia.The average serum UA levels,urine pH value of patients with dyslipidemia were 392μmol/L and 5,which were 339μmol/L and 6 in patients with normal lipid level.Significant difference were seen between the two groups ( P<0.05 ).Among 193 patients who had stone composition analysis, 130 cases had normal lipid level, accounting for 67.4%; 63 cases had dyslipidemia, accounting for 32.6%.In 63 patients with calculi who had dyslipidemia,31 cases had uric acid calculi,accounting for 49.2%.In 130 patients with calculi who had normal lipid level,40 cases had uric acid calculi, accounting for 30.8%.Significant difference were seen between the two groups ( P =0.013 ).Multivariate logistic regression showed TG was the independent risk factor of urinary stone formation ( P=0.001).Conclusion Dyslipidemia is closely related to urinary stone formation,especially concerning the for hypertriglyceridemia.