Effects of Bone Marrow Stromal Cells Transplantation Combined with Ultrashort Wave Therapy on the Expression of GFAP and ED-1 after Spinal Cord Injury in Rats / 中国医科大学学报

Objective To investigate the effects of bone marrow stromal cells(BMSCs)transplantation combined with low dose ultrashort wave (USW)radiation on functional recovery and the expression of glial fibrillary acidic protein(GFAP)and ED?1 after spinal cord injury(SCI)in rats,and further discuss its action mechanism. Methods Female Sprague?Dawley rats(n=30)were randomly divided into 5 groups:sham?oper?ated,as well as control,USW,BMSCs,and USW+BMSCs that were subjected to spinal cord injury(SCI). Basso?Beattie?Bresnahan(BBB)tests were carried out before the operation and at 1 d,1 week,2 weeks,3 weeks,4 weeks after SCI. 4 weeks later,animals were sacrificed and tissues were collected to make paraffin section. Immunohistochemical staining was performed to observe the expression of GFAP and ED?1. Results 4 weeks after SCI,BBB scores were significantly higher in the USW and USW+BMSCs groups than in the control group(both P<0.001). No signifi?cant difference was observed between the BMSCs group and control group. On the expression of GFAP ,only USW+BMSCs group showed signifi?cantly decreased compared with the control group(P<0.05). All treatment groups exhibited lower ED?1 expression than the control group(all P<0.05). Conclusion Our results indicate that USW radiation alone can obviously improve neural functional recovery after SCI. The USW radi?ation and BMSCs transplantation treatment can reduce inflammation ,and USW radiation is more effective. The combination therapy did not show a synergistic action on promoting functional recovery ,but do have an effect on reducing the inflammatory response and glial scar formation.

Effect of Tetramethyl Pyrazine on Infarct Areas and Expressions of ED1,IL-1?,TNF-? after Focal Cerebral Ischemia Reperfusion in Rats / 中国中医药信息杂志

Objective To investigate the effect of Tetramethyl Pyrazine(TMP) for treating cerebral infarct by observing infarct areas,and to research its function mechanism by observing the changes of the expressions of ED1,TNF-? and IL-1? in the infarction region after focal cerebral ischemia reperfusion injury.Methods Sixty SD male rats in healthy condition were randomly and averagely divided into normal group,model group and TMP high,middle and low dose group.The cerebral infarct animal model was reproduced by modified thread-tie method.TMP high dose group was administered TMP 140 mg/kg,middle dose group was 120 mg/kg and low dose group was 100 mg/kg by intravenous.Then all the rats were killed.Thirty rats were taken to evaluate whether TMP can decrease the ratio of cerebral infarction areas in the TTC-reacting and formalin fixation brain tissue slices taken from the above-mentioned killed rats,with Pathology Image Analysis System.The masculine of ED1,IL-1? and TNF-? in the immuno-reacting cells in cerebral infarction brain tissue slice taken from other thirty killed rats were observed.Results TMP groups decreased the ratio of cerebral infarction areas,and also decreased ED1,TNF-? and IL-1? in immuno-reacting cell.Conclusion TMP can decrease the formation of cerebral infarction caused by cerebral ischemia reperfusion and its effect is possibly related to the activation of microglia,as well as the immuno-positive expressions of TNF-? and IL-1?.

Expression of CD70 during Thymus Regeneration in the Rat / 대한해부학회지

The thymus is the central lymphoid organ for development of bone marrow-derived precursor cells into mature T-cells. The thymic stroma provides the specialized microenvironment for the proliferation, differentiation, and maturation of the immature T-cells, thymocytes. The CD27 is a tumor necrosis factor (TNF) receptor family member whose expression is limited to cells of the lymphoid lineage. CD70, the ligand of CD27, is a TNF related trans-membrane protein induced upon activation on T and B cells, dendritic cells and macrophages. CD70/CD27 interaction plays a key role in T dependent B cell responses and is responsible for plasma cell differentiation. This study was performed to investigate the expression of CD70 during regeneration following acute involution induced by cyclophosphamide (CY) in the rat thymus using RT-PCR analysis and single and double immunohistochemistry. The results from RT-PCR analysis showed that CD70 is expressed in mouse thymic medullary interdigitating (IDC)-like cells (MDC), DC2.4 and Raw264.7 but not expressed in the mouse thymic epithelial cells (subcapsular/cortex epithelial cells, cortical epithelial cells and medullary epithelial cells). Interestingly, upregulation of CD70 expression was observed in the thymic stromal cells and thymocytes isolated from rat thymus during thymus regeneration. Furthermore, immunohistochemistry demonstrated that CD70 was mainly expressed in the ED1 positive macrophages predominantly in the thymic cortex both in the normal thymus and during thymus regeneration. In line with the data obtained by biochemical analysis, CD70 immunoreactive macrophages is increased both in number and in size during thymus regeneration. Thus, the results of the present study suggest that CD70 expressed on the thymic macrophages could play a role in the development of new T cells to replace T cells damaged by cyclophosphamide treatment during thymus regeneration.

Morphological Changes of Anterior Horn Cells of Lumbar Spinal Cord after Anterior Root Avulsion in Adult Rat / 대한해부학회지

The morphological changes in the anterior horn of the L4 and L5 spinal segments were observed following anterior root avulsion in the adult male Sprague-Dawley rat (300~350 gm) at 5 days, 1 week, 2 weeks and 3 weeks postlesion. The animals were perfused with 4% paraformaldehyde, 0.15% picric acid in 0.1 M phosphate buffer solution and cryostat sections were prepared. Immunohistochemistry was used to identify changes of the phenotype in the anterior horn cells. Primary antibodies, goat anti-choline acetyltransferase (ChaT, 1 : 500, Chemicon), mouse antirat ED-1 (1 : 200, Serotec), rabbit anti-glial fibrillary acidic protein (GFAP, 1 : 200, DAKO) and rabbit anti-vascular endothelial growth factor (VEGF, 1 : 500, Santa Cruz Biotechnology) were used. Avidin-Biotin complex method was performed for immunohistochemical reaction and color reaction was developed with DAB-H2O2. Following results were observed in the anterior horn of lumbar spinal cord; 1. The number of ChaT-immunoreactive (ir) cells were reduced 20% level of control animals at 3 weeks after avulsion. 2. ED-1-ir microglia were significantly increased at 1 week and processes of ED-1-ir microglia surrounded around the axotomized neuronal cell bodies. 3. Gliosis defined by extensive GFAP immunoreactivity was observed both ipsilateral and contralateral side of lesion but the VEGF-ir cells were significantly increased in the ipsilateral side of lesion. Therefore, this study suggested that the majority of axotomized motor neurons were degenerated and the cellular proliferation and phenotype changes including glial cell activation were observed in the lumbar spinal cord after anterior root avulsion of adult rats.

The anti-inflammatory effects of angiotensin II blockade in animal model of cyclosporine-induced nephropathy / 대한내과학회지

BACKGROUND: Long-term treatment of immunosuppresant CsA causes interstitial inflammation and fibrosis in the kidney. Renin-angiotensin system (RAS) plays the most important role in the pathogenesis CsA-induced renal injury. Accordingly we evaluated the anti-inflammatory effect of angiotensin II blockades using losartan (LSRT) in a rat model of chronic CsA nephropathy. METHODS: Male Sprague-Dawley rats, initially weighing 225 to 250 g, were used. After 1 week of a low-salt diet (0.05% sodium), the rats were randomized into four groups and treated for 4 weeks. The Vehicle (VH) group was treated with olive oil. The VH+LSRT group was treated with olive oil and LSRT. The CsA group received CsA. The CsA+LSRT group was simultaneously treated with CsA and LSRT. The anti-inflammatory effect of LSRT was evaluated with C-reactive protein (CRP) expression, osteopontin (OPN) mRNA and protein expression, and ED-1 infiltration RESULTS: The CsA treatment caused an increase in serum creatinine and a decrease in creatinine clearance compared with that of the VH group. Intrarenal CRP positive cells were significantly decreased in the CsA+LSRT group compared with the CsA group (38.0 +/- 2.1 vs. 65.0 +/- 5.1, p<0.01). In the CsA group, the degree of OPN mRNA expression was increased compared with that of the VH group. But, OPN mRNA expression was decreased in the CsA+LSRT group (387.5 +/- 56.6% vs. 719.8 +/- 58.5%, p<0.05). In the degree of ED-1 infiltration, we had a similar results such as CRP and OPN mRNA expression (CsA group 30.5 +/- 8.0 vs. CsA+LSRT 86.0 +/- 11.0, p<0.01). CONCLUSION: We concluded that the anti-inflammatory effects of angiotensin II blockade has a potential protective effect against CsA-induced renal injury.

Mutations in ED1 Gene of Two Pedigrees with X-linked Hypohidrotic Ectodermal Dysplasia / 中华皮肤科杂志

Objective To detect ED1 gene mutations in the families with X-linked hypohidrotic ec-todermal dysplasia (XLHED). Methods Blood samples were obtained from 2 pedigrees. All 8 exons and flanking intronic boundaries of ED1 gene were amplified with polymerase chain reaction technique and then directly sequenced. Results Two mutations were found in ED1 gene. One was splicing mutation (IVS8+5 del G), the other was missense mutation (A959G). None of the mutations was found in normal individuals of two XLHED families and in 188 unrelated, population-matched control individuals. Conclusion Out of the ED1 gene mutations identified in 2 Chinese XLHED families, IVS8+5del G is a novel mutation.

Kidney-Specific Regulation by Angiotensin II / 대한신장학회잡지

Angiotensin II(ANG II) plays an important role in the regulation of systemic and renal hemodynamics, and functions as a growth factor in various tissues. ANG II is also known to be associated with healing after tissue injuries as profibrotic molecule. To determine effects of ANG II itself on kidney and to know its mechanism of action, 36 adult male Sprague-Dawley rats were infused with ANG II at a dose of 50ng/min(low ANG group), 100ng/min(high ANG group), or vehicle(control group) for 1 week using osmotic minipump(Model 2001D, Alza Co, USA) inserted into the interscapular area of the back. Tissue fibrosis was quantitated morphometrically using point detection method after Masson-Trichrome stain. Expression of ED-1 was determined by immunohistochemical staining. Immunohistochemical stain, RT- PCR, and Western blot assay were done for TGFbeta1 mRNA and protien. Results were as follows:1) There were no differences in body weight or kidney weight/body weight among 3 groups. 2) Interstitial volume was increased significantly in the low and high ANG groups compared with the control group(P<0.05) 3) ED-1 positive cells were increased significantly in the the low and high ANG groups compared with the control group(P<0.05). 4) TGFbeta1 mRNA and protein expression were increased significantly in the low and high ANG groups compared with the control group(P<0.05). Therefore, regardless of systemic hypertension, ANG II infusion induced renal fibrosis and increased expression of TGFbeta1 mRNA and protein in the kidney of the Sprague-Dawley rat.