ABSTRACT
To isolate bone marrow mesenchymal stem cells (BM-MSCs) and establish the model of chronic kidney disease (CKD) of Wuzhishan (WZS) mini-pig, and to study the repairment effect of BM-MSCs on CKD-induced renal fibrosis in vitro. Methods: Density gradient method was used to isolate and culture BM-MSCs. The cells were verified by morphology, phenotype, differentiation and so on. The left partial ureteral obstruction (LPUUO) was used to establish the CKD model, which was evaluated by B-ultrasound, single-photon emission computed tomography (SPECT), HE and Masson staining. The cells were divided into 3 groups, the tissue plus BM-MSCs group, the tissue group, and the BM-MSCs group, respectively. Seven days later, the supernatants were collected to observe the changes of hepatocyte growth factor (HGF) cumulative release. HE and Masson staining was used to observe the changes of renal tissue. Results: The isolated BM-MSCs possessed the features as follow: fibroblast-like adherent growth; positive in CD29 and CD90 expression while negative in CD45 expression; osteogenic induction and alizarin red staining were positive; alcian blue staining were positive after chondrogenic induction. Twelve weeks after the operation of LPUUO, B-ultrasound showed the thin renal cortical with pelvis effusion; SPETCT showed the left kidney delayed filling and renal impairment. The accumulation of HGF in the tissue plus BM-MSCs group was significantly higher than that in the tissue alone group at the 1st, 5th, 6th, 7th day, respectively (P<0.05). HE staining showed the different degree of renal lesions between the tissue plus BM-MSCs+CKD group and the tissue alone group, which was aggravated with the time going. Masson staining showed that the cumulative optical density of blue-stained collagen fibers in tissue plus BM-MSCs group was significantly lower than that in the tissue group at the 5th to 7th day (P<0.05). Conclusion: BM-MSCs from WZS mini-pig can inhibit or delay the progress of CKD-induced renal fibrosis through autocrine HGF in vitro.
Subject(s)
Animals , Autocrine Communication , Physiology , Bone Marrow Cells , Cells, Cultured , Fibrosis , Hepatocyte Growth Factor , Metabolism , Kidney , Pathology , Mesenchymal Stem Cells , Renal Insufficiency, Chronic , Swine , Swine, Miniature , Ureteral ObstructionABSTRACT
OBJECTIVE@#To establish a method to isolate, culture and identify bone marrow-derived mesenchymal stem cells (BM-MSCs) from inbreed line miniature pig of Wuzhishan (ILMW) in vitro, to compare the biological characteristics of BM-MSCs derived from different pigs, and to supply BM-MSCs for investigating the repair mechanisms of renal injury in ILMW aft er unilateral ureteral obstruction.@*METHODS@#Four or 10-months old ILMW (n=4 per group) were selected and 5 mL of bone marrow fluid was extracted at 1 cm position from iliac wing edge. Mononuclear cells were isolated by density gradient method, then were cultured in the complete medium containing 3 different kinds of fetal bovine serum (FBS) (10% FBS, 12% FBS or 15% FBS). The cells of Passage 1, Passage 3, Passage 5 or Passage 11 were collected to examine biological characteristics including morphology, phenotype, differentiation ability, growth curve and cell cycle.@*RESULTS@#The BM-MSCs were attached to the culture dishes, which were fibroblast-like or whirlpoollike. Primary cultured cells began the adherence at 18 h and entered a logarithmic phase in the 6th day. Eighty percent of them were fused in the 9th day. There were no obvious anomalies in the subcultured cells. The expressions in cell surface antigens of CD29, CD44 and CD90 were positive, while the expressions of CD34 and CD45 were negative. There was no statistically significant difference between cells from different generations (all P>0.05). Under condition of osteogenic induction, alizarin red staining was positive at the 18th day, and alcian blue staining was positive at the 21th day. Cell cycle examination showed that the rate of G0/G1 was about 81.45%.@*CONCLUSION@#BM-MSCs of ILMW has advantages of earlier adherent time, more active proliferation, shorter cell subculture cycle, and stable biological characteristics after subculture, which is one of the best kinds of BM-MSCs coming from swine in mainland of China.
Subject(s)
Animals , Cell Differentiation , Cell Division , Cell Separation , Cells, Cultured , China , Mesenchymal Stem Cells , Cell Biology , SwineABSTRACT
0.05),and the costs were 330.27 yuan,540.48 yuan and 790.73 yuan,respectively. CONCLUSION: Regime A was proved to be the optimal scheme.
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OBJECTIVE:To evaluate the cost-effectiveness of amiodarone versus sotalol for Atrial fibrillation.METHODS:A total of 102 patients with atrial fibrillation were treated with amiodarone(Group A)or sotalol(Group B).The cost-effectiveness analysis was performed on two groups.RESULTS:The total cost in Group A was 2 714.18 yuan versus 2 176.56 yuan in Group B;the effective rates were 78.43% vs.70.59%(P
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OBJECTIVE:To study the incidence,the etiological factor and the harm of metoclopramide-induced children poisoning.METHODS:We reviewed 30 reports published in Chinese Pharmaceutical Abstracts(2003～2006)and domestic pediatric journals and gave a comprehensive evaluation on the confirmed 56 metoclopramide-induced children poisoning cases.RESULTS:All of the 56 poisoning cases were induced by oral administration and all had high daily dose.The condition was predominantly characterized by posterior tilt of head,stiffing of root of tongue,ataxia,upward staring or side-glance etc.Metoclopramide has inhibition effect on leucocytes,and it can result in cumulative poisoning,especially for children with hepatorenal inadequacy.CONCLUSION:Great attention should be attached to the dose and course of treatment of metoclopramide to enhance drug efficacy while reducing incidence of adverse drug reactions.