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Objective:To investigate the effect and mechanism of bone marrow-derived mesenchymal stem cells (MSCs) in alleviating renal injury in unilateral ureteral obstruction (UUO) model.Methods:MSCs were cultured, and then transplanted into a UUO model through the tail vein. Histology, cell apoptosis, peritubular capillary (PTC) loss and β-catenin expression were examined on the fourteen day after surgery.Results:Renal interstitial fibrosis in the MSCs group was significantly attenuated compared with that in the UUO group (HE: 1.60 ± 0.35 vs. 3.34 ± 0.23; MASSON: 21.32 ± 7.54 vs. 51.08 ± 4.45). Moreover, MSCs treatment inhibited the loss of peritubular capillaries (PTC) (13.56 ± 4.65 vs. 60.16 ± 10.24), cell apoptosis (14.32 ± 3.54 vs. 28.16 ± 6.21) and β-catenin expression (29.33 ± 6.45 vs. 39.51 ± 8.42).Conclusions:MSCs infusion is a promising therapeutic strategy for promoting kidney repair in chronic renal fibrosis model. The mechanism maybe that it inhibits the loss of peritubular capillaries (PTC) , cell apoptosis and β-catenin expression.
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In recent years, more and more studies have shown that myeloid-derived suppressor cells (MDSCs) participate in the development and progression of various chronic liver diseases including chronic viral hepatitis, alcoholic liver disease, nonalcoholic fatty liver disease, autoimmune liver diseases, and liver cancer. As a type of cells derived from bone marrow progenitor cells and immature myeloid cells, MDSCs play an important role in the development, progression, and repair of liver diseases by regulating inflammatory response and the differentiation and function of immune cells. This article reviews the research advances in the association between MDSCs and various liver diseases, in order to provide new thoughts for the clinical diagnosis, prognosis, and treatment of chronic liver diseases.
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Objective@#To study the effects of nerve growth factor (NGF) on the proliferation, osteogenic differentiation and mineralization of type 2 diabetic mice bone marrow stromal cell (BMSC), providing basis for clinical application of NGF.@*Methods@#Three 8-week-old male db/db mice and two 8-week-old male C57BL/6J mice were used in the study. BMSC derived from femur were cultured though adherence method. BMSC of C57BL/6J mice and db/db mice was divided into normal group and diabetic group to conduct the osteogenic potential experiment, named experiment one. In experiment two, diabetic BMSC was divided into 3 groups: diabetic control group, NGF group, and K252a+NGF group [K252a was the inhibitor of tyrosine kinase A (TrkA), which was the high affinity receptor of NGF], to investigate effect of NGF on osteogenic potential of diabetic mice BMSC. After seeding BMSC, K252a was added into K252a+NGF group, then NGF was added 30 min later. NGF was added into NGF group and K252a+NGF group, but not diabetic control group. The proliferation of BMSC at 1, 3, 5 and 7 d in experiment one and the proliferation of BMSC at 1, 2 and 3 d in experiment two were evaluated through methyl thiazolyl tetrazolium, and the level of alkaline phosphatase (ALP) at 3, 5 and 7 d in both experiments were measured. After being osteogenic induced for 14 d, mineralized nodules in both experiments were quantitated by alizarin red calcium stain. Five holes were set in every group, and all experiments were repeated 3 times.@*Results@#The BMSC proliferation of diabetic group was significantly higher than that of the normal group at 3, 5 and 7 d (P<0.05). After being osteogenic inducted for 3, 5 and 7 d, ALP level of diabetic group were significantly lower than that of normal group (P<0.05). After being osteogenic inducted for 14 d, calcium nodule count of diabetic group [(23.1±6.4) nodule/field] were significantly lower than that of normal group [(36.9±7.9) nodule/field](P<0.05). At 1, 2 and 3 d, BMSC proliferations of diabetic control group, NGF group and K252a+NGF group were not statistically different (P>0.05). After being osteogenic inducted for 3 and 5 d, ALP level of NGF group was significantly higher than that of diabetic control group (P<0.05). After being osteogenic inducted for 3, 5, and 7 d, ALP level of K252a+NGF group was significantly lower than that of NGF group (P<0.05) and diabetic control group (P<0.05). After being osteogenic induced for 14 d, calcium nodule count of NGF group [(45.2±6.8) nodule/field] was significantly more than that of diabetic control group [(23.1±6.4) nodule/field](P<0.05); while calcium nodule count of K252a+NGF group [(18.0±4.5) nodule/field] was significantly less than that of NGF group (P<0.05) and diabetic control group (P<0.05).@*Conclusions@#The differentiation and mineralization of type 2 diabetic mice BMSC was significantly reduced. NGF promoted the osteoblastic differentiation and mineralization of diabetic mice BMSC in viro though combining with TrkA.
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Objective To explore the in vitro culture methods for oriented differentiation of peritoneal cells and bone marrow cells into high-purity mast cells,and to identify the function of these mast cells.Methods Peritoneal cells and bone marrow cells were isolated from the peritoneal cavity lavages and femur of C57BL/6 mice,and cultured with both interleukin-3 (IL-3) and stem cell factor for 2 and 4 weeks respectively.Light microscopy was performed to observe the morphology of these cells,toluidine blue staining to identify the degree of maturity of these mast cells,and flow cytometry to measure the expression of cell surface markers C D 117 and FceR Ⅰ α.After the stimulation with compound 48/80 at different concentrations,the degranulation rate of mast cells was counted under the microscope,and β-hexosaminidase release rate was measured by spectrophotometry.Results After 2-or 4-week culture,the mouse peritoneal and bone marrow cells all manifested as refractive suspension cells of uniform size.Toluidine blue staining showed violaceous metachromatic granules in the cytoplasm of the two kinds of cells.The proportions of CD117 or FcεR Ⅰ α single-positive peritoneal and bone marrow-derived mast cells were all more than 95%,and the proportions of CD117/FcεR Ⅰ α double-positive peritoneal and bone marrow-derived mast cells were 97.68% ± 0.80% and 96.12% ± 0.76% respectively.The degranulation rates of mast cells in the 100-and 1 000-mg/L compound 48/80 groups significantly differed from those in the blank control group (all P < 0.01).Compared with the blank control group,the β-hexosaminidase release rates significantly increased in bone marrow-derived mast cells in the 100-mg/L compound 48/80 group and peritoneal mast cells in the 10-and 100-mg/L compound 48/80 groups (P < 0.01 or 0.05).Conclusion IL-3 and stem cell factor can co-induce the directed differentiation and proliferation of mouse bone marrow stem cells and peritoneal cells,so as to harvest highnuritv mature degranulated mast cells,and lay a foundation for subsequent cell biology research.
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Objective To investigate the protective effect of indole-3-carbinol (I3C) on radiation-induced mouse bone marrow hematopoietic cell injury and the involved mechanisms. Methods (1) The bone marrow nuclear cells (BMNCs) from CD45.1 subtype of C57BL/6J mice were collected by a density gradient centrifugation method. The BMNCs were pretreated with a series doses of I3C (0 mol/L, 10-8 mol/L-10-3 mol/L) and then exposed with radiation of 137Csγ-ray (doses of irradiation were 0 Gy, 1 Gy and 4 Gy). After 18-hour culturing, the bioluminescence method was used to detect the cell viability. (2) These cells were divided into control group and 10-6 mol/L I3C group. Both groups were received the irradiation (0 Gy, 1 Gy and 4 Gy) and inoculated into the methylcellulose semi-solid culture medium to incubate 7 days, the colony forming unit-granulocyte monocytes (CFU-GM) were observed. (3) Twenty-four CD45.2 subtype mice used as the receptor were exposed with 8 Gy radiation. The CD45.1 BMNCs were divided into control group, 4 Gy irradiation group, 4 Gy irradiation and 10-6 mol/L I3C group. Donor cells were harvested from C57BL/6J (CD45.1) mice after they received various treatments, and were then mixed with competitive BMNCs from C57BL/6J (CD45.2) mice. The mixed cells were transplanted into recipient mice (8 mice/group). Flow cytometry was used to analyze the proportion of donor cells in peripheral blood of receptor. (4) The cells were divided into control group, 10-6 mol/L I3C group, 1 Gy irradiation group, 1 Gy irradiation with 10-6 mol/L I3C group. After 24-hour culturing, Western blot assay was used to detect the expression levels of nuclear factor erythroid 2-related factor 2 (Nrf2) and hemeoxygenase-1 (HO-1). Results (1) I3C showed a significant cytotoxic effect on the BMNCs when its concentration was above 10-4 mol/L. 10-7-10-6 mol/L I3C could reduce the radiation injury of BMNCs under the same dose of irradiation. Therefore, 10-6 mol/L I3C was chosen for subsequent experiments. (2) The CFU-GM was significantly higher in 10-6 mol/L I3C group than that of control group (P<0.05). (3) Results of flow cytometry showed that the proportion of donor cells in receptor was significantly higher in 4 Gy irradiation group than that of control group, which decreased the engraftment capability of irradiated HSCs (P<0.05), although the engraftment capability of irradiated HSCs improved after 10-6 mol/L I3C treatment. (4) I3C significantly enhanced the increased protein expression of Nrf2 and HO-1 caused by radiation (P<0.05). Conclusion I3C has a protective effect on hematopoietic cells following radiation-induced injury, which may be related with activating the Nrf2/HO-1 signal pathway.
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Current researches indicate that transplanted BM‐MSCs accompanies massive death ,and its effect is unsat‐isfactory .The miRNAs are a class of small non‐coding single‐stranded RNA molecules ,involve therapeutic course of bone marrow mesenchymal stem cell treating myocardial infarction in many respects .The present article made an o‐verview on effect of microRNAs in transplanted BM‐MSCs treating myocardial infarction via mainly illustrating bio‐logical function of microRNAs and its regulation effect on BM‐MSCs differentiation and paracrine secretion .
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BACKGROUND:The assessment for long-term efficacy of chronic ischemic disease is more important than the short-term efficacy assessment, which associates with patient’s long-term quality of life and long-term survival rate. OBJECTIVE:To observe the 5-year folow-up outcomes of autologous bone marrow stem cel transplantation for the treatment of thromboangitis obliterans. METHODS:This study enroled 43 patients of thromboangitis obliterans who underwent autologous bone marrow stem cel transplantation from August 2007 to January 2010 in the Department of Thyroid Vascular Surgery, the First Affiliated Hospital of Xinjiang Medical University. At 1, 2, 3, 4 and 5 years after transplantation, pain, cold sensation, and intermittent claudication distance were folowed up by telephone; changes in limb ulcers were observed. At 1 year after transplantation, venous oxygen partial pressure and oxygen saturation of limbs were reviewed. RESULTS AND CONCLUSION:A total of 38 thromboangitis obliterans patients with complete folow-up data were included in the final analysis. Compared to the preoperation, pain, cold sensation, and intermittent claudication significantly improved. The difference was statisticaly significant (Z values:-4.277,-5.086,-3.574, P 0.05). Intermittent claudication distance had increased. Differences in terms of intermittent claudication distance was statisticaly significant (Z=43.898,P < 0.001). Significant differences in venous oxygen partial pressure and oxygen saturation were detected between preoperation and 1-year posttransplantation (tvalues: 36.790, 43.964,P values: 0.040, 0.037). Above results suggest that autologous bone marrow stem cel transplantation for thromboangitis obliterans obtained stable long-term outcomes.
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Objective To study characteristics and immune mechanisms of CD11b+ GR-1-myeloid-derived suppressor cells (CD11b+ GR 1+ MDSCs) in elderly mice,as compared with those of healthy young mice.Methods Totally 20 healthy C57BL/6 young mice (aged 1-2 months) and 20 elderly mice (aged over 18 months) were randomly chosen and splenetic CD11b+ GR-1+ MDSCs were sorted with the MDSCs Isolation Kit.In vitro assays,the effects of young and elderly CD1 1b+ GR 1+ MDSCs on the proliferation of T cells were determined by Brdu Elisa.Transwell co-culture and real-timePCR were used to identify the mechanisms of different immune suppressive functions of CD11b+GR 1+ MDSCs sorted from young mice and elderly mice.Results Compared with young MDSCs,elderly MDSCs could evidently suppress the proliferation of T cells (t=8.67,P<0.001),and this function could be reversed by trans-well co-culture (t=6.93,P<0.001).The results of realtime PCR revealed that,compared with young MDSCs,elderly MDSCs expressed higher levels of arginase-1 (ARG-1),inducible nitric oxide synthase (iNOS),reactive oxygen species (ROS),interleukin 10 (IL-10),IL13 and transforming growth factor (TGF)-β (t=9.04,4.86,7.04,6.92,4.51,5.46,respectively,P<0.05 or P<0.01).Conclusions CD11b+GR-1+MDSCs sorted from healthy elderly mice can evidently suppress the proliferation of T cells through cell-cell contact and secretion of suppressive medium.
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Objective To explore the effect of astragalus extracts on adherence, migration, cell viability, microvascular formation, and eNOS expression in bone marrow-derived endothelial progenitor cells (EPCs) of rats. Methods EPCs were cultured, isolated and identified in vitro and divided into four groups: low titer group (10-4 g/L of astragalus extracts), middle titer group (10-3 g/L of astragalus extracts), high titer group (10-2 g/L of astragalus extracts) and control group. The effects of astragalus extract on adhesion, migration or microvascular formation were observed under the inverted microscope and com?pared between all groups;Cell viability was detected by MTT assay;mRNA transcription and protein expression levels of en?dothelial nitric oxide synthase (eNOS) in EPCs were detected by reverse transcription PCR (RT-PCR) and Western blot re?spectively. Results Compared with the control group, cell adhesion, migration and microvascular formation of EPCs all en?hanced with addition of astragalus extracts in a dose dependent manner (F=15.256, 13.633, 97.549 respectively, and P <0.05 in all cases); Cell vitality of EPCs increased with administration of astragalus extracts in a time and dose dependant manner. (F=9.755 for time and F=10.18 for dose). mRNA transcription and protein expression of eNOS in EPCs were up-reg?ulated with addition of astragalus extracts in a dose dependent manner (F=56.356 and 77.125 respectively, and P<0.05 in both cases). Conclusion Astragalus extracts play important role in angiogenesis in EPCs probably through up-regulating expressions of eNOS.
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Objective To evaluate the bone repair capacity ofpolylactic acid-polyglycolic acid copolymer (PLGA)/collagen type Ⅰ (CoI) microspheres combined with BMSCs after being injected in intertrochanteric bone defect of osteoporotic female rats.Methods Prepared PLGA microspheres.The microspheres were coated with Col.BMSCs of the third passage were cultured with PLGA/CoI microspheres.Forty 3-month-old female SD rats were ovariectomized to establish osteoporotic animal models.The osteoporotic rats were randomly divided into 5 groups,including SHAM group,PBS group,Cell group,MS group and Cell+ MS group.There were 8 rats in each group.Different material was injected into the intertrochanteric bone defect site which was made with electric drill.Four rats of each group were sacrificed at 1 month and 3 months post-operation.The fenora were taken to measure the intertrochanteric bone mineral density (BMD) with DEXA and evaluate trabecular stucture with Micro CT.Results After 7 days of coculture,BMSCs seeded on PLGA/CoI microspheres had nice adherance and proliferation.There was no difference of BMC and BMD among all groups at 1 month post-operation.Tb.Th of Cell+MS group was higher than that of PBS group and MS group at 1 month post-operation.%Tb.Ar of Cell+MS group was higher than that of Cell group and MS group at 1 month post-operation.Tb.Sp of Cell+MS group had a tendence to decrease compared with other groups but there was no statistical difference at 1 month post-operation.After 3 months of operation,the BMC of Cell+MS group had a tendence to increase compared with that of PBS group and MS group but showed no statistical difference.BMD and Tb.Th of Cell+MS group was higher than those of other groups.%Tb.Ar of Cell+MS group was higher than that of SHAM group and PBS group.Tb.Sp of Cell+MS group had a tendence to reduce compared with other groups but showed no statistical difference.Conclusion The bone defect of osteoporotic site can be repaired 1 month after the injection of the PLGA/CoI microspheres combined with BMSCs.The trabecular reconstruction and bone quality of osteoporotic site can be improved 3 months after the injection.
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BACKGROUND:Epimedium has been used in the treatment of osteoporosis and repair of bone defects. OBJECTIVE:To explore the effects of epimedium on the osteogenic differentiation of bone marrow mesenchymal stem cells. METHODS:A database search was performed to retrieve literatures addressing epimedium effects on the osteogenic differentiation of bone marrow mesenchymal stem cells. Then, the papers meeting the criteria were selected for in-depth analysis. During the osteogenic differentiation induced by epimedium, alkaline phosphatase, osteocalcin, osteopontin, transforming growth factorβ, bone morphogenetic proteins, osteocalcin, bone sialoprotein were detected in the bone marrow mesenchymal stem cells to understand the underlying mechanism of epimedium in proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells. RESULTS AND CONCLUSION:Epimedium effects on bone marrow mesenchymal stem cells are shown in a dose-dependent manner. During the early induction, icari n can increase cellphosphatase activity;in the late induction, icari n can increase calcified nodules, promote osteocalcin secretion, significantly improve the expressions of transforming growth factorβ1, bone morphogenetic protein 2, insulin-like growth factor-1, osteopontin and bone sialoprotein. Epimedium, which can be used as an excellent osteoinductive factor, improves the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells.
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BACKGROUND:celltherapy by the implantation of autologous bone marrow cells has been used for the treatment of ischemic heart diseases in clinical trials for decade. However, as the outcomes of celltransplantation obviously vary among patients, it is essential to identify the risk factors that may influence the level and function of progenitor cells in bone marrow, in order to identify the patients who would benefit the most from this treatment. OBJECTIVE:To observe the impact of perioperative cardiovascular risk factors on number and function of bone marrow progenitor cells from patients undergoing coronary artery bypass grafting surgery. METHODS:We col ected clinical and laboratory data from 44 patients scheduled to undergo sternotomy for coronary artery bypass grafting procedures. Bone marrow was aspirated from the sternum during the operation and bone marrow mononuclear cells were isolated by density centrifugation with Ficol lymphoprep and then detected using trypan blue exclusion method. Levels of progenitor cells in bone marrow were evaluated using flow cytometry. Function of bone marrow progenitor cells were assessed by clonogenic and migration assays. RESULTS AND CONCLUSION:We assessed the number of bone marrow mononuclear cells out of 20 mL bone marrow in duplicate samples from patients with coronary heart disease scheduled for coronary artery bypass grafting that was (10-89)×106 cells with over 95%activity. A negative correlation was observed between the number of bone marrow mononuclear cells and the age (n=44, r=-0.788, P=0.001). Levels of CD34+, CD133+, and CD34+CD133+cells in bone marrow mononuclear cells was (0.94±0.39)%, (0.46±0.28)%, and (0.53±0.26)%. Levels of CD34+cells and CD133+cells in patients with diabetes were significantly lower than those in patients without diabetes. Female, advanced age and poor heart function were related with reduced colony-forming ability of progenitor cells. A positive correlation was observed between level of CD34+cells and migration ability of bone marrow mononuclear cells. The results show that by density gradient centrifugation, we can harvest a sufficient number of bone marrow mononuclear cells in the treatment for ischemic heart disease. Age, gender, diabetes, heart function are correlated with bone marrow mononuclear cellnumber and functions.
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Tumor invasion and metastasis are regarded as main reasons for the failure of therpy and the reason of patients death.The mechnism of tumor metastasis is still uncertain.The pre-metastatic niche hypothesis provides us with new ideas to discover the mechnism.Numerous materials are involved in the formation of the pre-metastatic niche according to this hypothesis,including bone marrow-derived cells,microvesicles,exosomes,CD44,and so on.A further research on this hypothesis helps to deeply understand the nature of metastasis and leads clinical doctors to explore novel targets for clinical diagnoses and therapies.
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Objective To observe the effects of transplantation of bone marrow stem cells(BMSCs)transfected with bone morphogenetic protein-2(BMP-2)on fracture healing in rats with diabetes so as to provide a new therapy for diabetic fractures.Methods Fifty male adult Wistar rats aged six weeks randomized to the control group and experimental group were all employed to establish models with diabetic fractures.Under high glucose condition,BMSCs were transfected with BMP-2 by adenovirus vector in vitro.BMSCs transfected by BMP-2 were transplanted into the fracture area of rats in the experimental group,while non-transfected BMSCs into the corresponding area of rats in the control group.X-ray examination was performed at 1,2,3,4 and 6 weeks after transplantation.Bony calluses were collected for HE staining and gray scales of BMP-2 in calluses were determined by immunohistochemical method.Meanwhile,serum levels of BMP-2 were measured by ELtSA.Results The gray scales of BMP-2 in the calluses were 83±3 in the experimental group and 118±4 in the control group at four weeks(P<0.01).The serum concentrations of BMP-2 were(203.80±8.96)ng/L in the experimental group and(139.15±4.19)ng/L in the control group at four weeks(P<0.01).Conclusion Transplantation of BMSCs transfected by BMP-2 promotes fracture healing in diabetic rats.
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ObjectiveTo explore the effects of bone morrow stromal cells (BMSCs) on the neurological behavior of rats with traumatic brain injury (TBI).MethodsTwenty-four SD rats were randomly and equally divided into control group, TBI group and BMSC group. The weight-drop device was adapted to establish the TBI model. The injury severity and its outcome were evaluated by a set of criteria termed neurological severity score (NSS). Brain tissues were harvested at day 14 to observe the survival and migration of the transplanted cells.Bax expression was detected by RT-PCR. Results NSS was (12 ±3 ) points in the TBI group, significantly higher than (7 ± 1 ) points in the BMSC group (P <0.05). The transplanted BMSCs could survive and migrate. Moreover, BAX, a crucail apopotosis gene, was down-regulated to 0.9 ±0.1 in the BMSC group, compared with 1.1 ±0.2 in the TBI group (P <0.05). ConclusionsBMSC transplantation is available to improve the neurological function, as may be associated with the Bax.
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Objective To investigate the amounts of endothelial progenitor cells(EPCs)in peripheral blood of patients with proliferative diabetic retinopathy(PDR).Methods Forty patients with PDR(PDR group),thirty patmnts with type 2 diabetes mellitus(DM)without DR(DM group),and twenty agematched normal subjects(control group)were enrolled in this study.Blood samples were treated bv repeated centrifugation and stained with monoclonal antibodies.At least 2 × 105 cells were analyzed bv flow cytometry.EPCs were identified by CD34 and CD133 antibody.The correlation between EPCs numbers and DR duration,glycosylated hemoglobin,serum lipids was analyzed.Results The number of EPCs in PDR,DM and control group were(49±12)、(35±11)、(90±25)cells/ml respectively,the difference was statistically significant(F=56.260,P=0.000).There was a positive correlation between EPCs numbers and DR duration(r=0.564,P<0.05).However there was no correlation between EPCs numbers and glycosylated hemoglobin(r=-0.170,P>0.05)or triglyceride levels(r=0.261,P>0.05).Conclusions The number of EPCs in peripheral blood of PDR patients was decreased. EPCs might play an important role in the pathogenesis of PDR.
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Objective To observe the effect of diabetic retinopathy on endothelial progenitor cells (EPCs)from peripheral blood.Methods Sixty male Wistar rats were divided into control group and diabetes group.The rats in diabetes group were induced with streptozotocin(STZ)injection for diabetic retinopathy model.Flow cytometry was used to identify and count the number of EPCs from peripheral blood at 1 week.1,3 and 6 months after injection.All eyeballs were examined by hematoxylin and eosin (HE)staining,periodic acid-Schiffs(PAS)staining of trypsin-digested retinal vessels flat preparation and transmission electron microscope.EPCs count,and the relationship between DR morphological changes and EPCs count were compared and analyzed.Results The quantity of EPCs from peripheral blood at 1 week,1,3 and 6 months after STZ injection were 25±7,28±8,39±7,43±7 cells per 200 000 monocytes respectively,which decreased compared with the control group 45±4 cells per 200 000 monocytes(F=8.933,P<0.0 1).The quantity of EPCs was gradually increased at 1 week,1,3 and 6 months after STZ injection,accompanied with responsive pathological changes of retinal structure and vessels.The thickness of retina at 1 week and 1 month after injection were reduced slightly.The number of retinal ganglion cells reduced,with the time passing by.Endothelial cells were edema,mitochondrial was swollen,capillary basement membrane was thicken,lumen was significant stenosis,lumen occlusion and retinal artery aneurysm were observed at 6 months after STZ injection.Conclusion The number of EPCs increases gradually throughout the development of DR.
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Objective To evaluate the efficacy of MRI for assessment of re-distribution of bone marrow mesenchymal stem cells injected intramyocardially in main organs (heart, liver, spleen and kidney) under different heart status (beating or arresting) in a porcine model. Methods Bone marrow-derived mesenchymal stem cells were obtained from the male swine and labeled with iron oxide during culture. Acute myocardial infarction was created in female swine, one week later, the survivors were randomly divided into 4 groups. Cardiopulmonary bypass was set up to arrest the heart, and then labeled cells (1×108) were intramyocardially injected into the border of the infracted myocardium in group 1 (n=6). The same volume of cells was grafted into the beating heart in group 2 (n=6). In group 3 and 4, saline was injected into either the arresting or beating myocardium. Three days later, re-distribution of stem cells and cardiac function were assessed by T2*WI and cine MRI, respectively. All animals were sacrificed for histology and real-time quantitative polymerase chain reaction (RT-PCR) of sex-determining region on Y-chromosome (SRY) investigation.The ANOVA and t test was used for statistics. Results The left ventricular end-diastolic volume (LVEDV) before transplantation for group 1-4 were: (56.8±5.3),(54.8±6.8),(57.4±4.3)and(56.8±2.8) ml, and after transplantation for group 1-4 were: (65.2±5.2),(63.2±3.7),(60.2±4.7)and(62.2±4.4) ml. The left ventricular end-systolic volume (LVESV) before transplantation for group 1-4 were: (33.5±7.6),(32.3±5.3),(33.5±3.6)and(32.7±4.6) ml,and after transplantation for group 1-4 were: (37.3±5.6),(36.3±6.9),(34.3±5.4)and(36.3±8.1) ml. The left ventricular EF values (LVEF) before transplantation for group 1-4 were: (42.3±7.2)%,(41.7±6.8)%,(41.8±8.6)% and(42.7±7.7)%,and after transplantation for group 1-4 were: (44.5±8.7)%,(43.1±7.4)%,(42.8±5.6)% and(43.3±8.4)%. The myocardial infarction area (MI) before transplantation for group 1-4 were: (6.5±2.1),(6.4±1.9),(6.5±2.5)and(6.4±2.6) cm2,and after transplantation for group 1-4 were: (6.4±2.3),(6.2±2.6),(6.3±2.5)and(6.4±2.8) cm2 . There were no statistical differences before and after transplantation in these 4 groups[P values of before and after transplantation for LVEDV, LVESV, LVEF,MI were >0.05 (F= 0.277, 0.066,0.066, 0.003); and >0.05 (F= 1.137,0.182,0.021,0.008),respectively]. The T2 value of the infracted myocardium in group 1 decreased more obviously than that in group 2[(-22.3 ± 2.2) vs (-17.0 ± 0.8) ms, t=-5.489, P<0.01], while the T2 value of the spleen decreased more significantly in group 2 than that in group 1[(-7.7 ± 0.7) vs (-13.3 ± 1.1) ms,t=9.055, P<0.01]. The T2 values of the liver and kidney were no significant differences in group 1 and 2 (liver, t=-0.532,P>0.05 and kidney, t=-0.113,P>0.05). The results of RT-PCR in group 1 and 2 showed significant differences in heart[(150±62) vs (72±4) U/L ,P<0.05, t=3.109], spleen[(131±1) vs (233±17) U/L, P<0.01, t=- 13.286]and liver[(17±1) vs (9±5) U/L ,P<0.01,t= 3.492]. Pathological examination demonstrated that the transplanted stem cells were positive for Prussian blue staining, which had a good correlation with MRI results. Conclusion MRI can serve as a convenient and efficient imaging method to track the migration of stem cells with SPIO labeled in early stage and evaluate its early re-distribution in vivo. Injection of bone marrow mesenchymal stem cells in the arresting heart could favor retaining more cells in the myocardium.
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Objective To investigate the expression and clinical significance of P-selectin (CD62P) in bone marrow hematopoietic stem cells of patients with acute leukemia (AL). Methods The CD62P expression in bone marrow mononuclear cells of 15 healthy donors and 56 untreated patients with AL, were examined by flow cytometry. Results The average rate of CD62P expression was (6.72±7.64) % in hematopoietic stem cells (CD+45 CD+34 CD-38) of the 38 patients with acute myeloid leukemia (AML), was (3.46±2.51) % in hematopoietic stem cells (CD+45 CD+34 CD+19) of the 12 patients with B-acute lymphoblastic leukemia (B-ALL), and was (6.23±4.95) % in hematopoietic stem cells (CD+45 CD+34 CD+7) of 6 patients with T-acute lymphoblastic leukemia (T-ALL). The expression rates in those AL patients were higher than that in the healthy controls (1.04 ±1.23) % (t = 2.847, 3.284, 3.091, respectively, P <0.01), while there was no difference between the control group and the group who reach CR after routine treatment (t =1.932, P >0.05). Furthermore, the leukocyte,hemoglobin and platelet count in CD+62P patients with AML and T-ALL were significantly higher than CD-62P ones (t =4.153, 8.095, 8.289, 7.235, 8.692, 9.832, respectively, P <0.05), but there was no significant difference between CD+62P and CD-62P patients with B-ALL (t =0.340, 1.142, 0.019, respectively, P >0.05).Conclusion The CD62P is one of the markers of platelet activation, and its expression varies in different types of AL. The CD62P in hematopoietic stem cells of AL could be regarded as a new sign for the leukemic stem cells, as well as a helpful prognostic indicator in treatment response assessment.
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Objective To investigate the impact of bone marrow mesenchymal stem cell transplantation on a rat model of experimental autoimmune uveitis (EAU) and analyze its immune regulatory mechanisms in vivo. Methods Eighteen Lewis rats were randomly divided into three groups: model control group, intervention group and normal control group, six animals in each group. Human retinal S-antigen peptide (HS-AgP35, 1 mg/ml) was mixed and emulsified with complete Freund's adjuvant and injected into hind foot pad of rats on the first and eighth day to establish the animal model of EAU. For bone marrow mesenchymal stem cell transplantation, 1 ml of cell suspension (2 × 106 cells/ml) was injected into tail vein of the intervention group rats on the first day when the emulsified S-antigen was injected. EAU manifestation, pathological change and IFN-γ level were evaluated and compared among those three groups after two weeks. Results No abnormal signs were found in the eyes of rats in normal control group. The manifestation grading of the intervention group (two rats at grade 0, three rats at grade 0.5, one rat at grade one) was significantly different from the model control group (one rat at grade one, one rat at grade two, three rats at grade three, one rat at grade four) (P=0. 015). The retina of rats in normal control group was ordinary under light microscope. The histopathological grading of the intervention group (one rat at grade 0, four rats at grade 0.5, one rat at grade one) and the model control group (four rats at grade three, two rats at grade four) was also statistically different (P<0. 01). Furthermore, the IFN-γ level in peripheral blood of the intervention group rats declined significantly compared to the model control group (t=9. 057 4, P=0. 01). Conclusions Bone marrow mesenchymal stem cells can inhibit EAU significantly,possibly by lowering the level of IFN-γ, thereby reduce the severity of uveitis and improve the condition of uveitis in rats.