ObjectiveTo observe the effect of rabbit skeletal muscle stem cells (RSMSCs) modified by adenovirus mediated bone morphogenetic protein-2 (BMP-2) gene ex vivo in combination with demineralized bone matrix (DBM) on repair of longer bone defect in rabbit.MethodsModel of radial bone defects (20 mm) of rabbits was established. 50 rabbits were divided into 5 groups, group A (AdrhBMP-2 trusduced RSMSCs/DBM group), group B (adGFP trusduced RSMSCs/DBM group), group C (not trusduced RSMSCs/DBM group), group D (DBM group), and group E (untreated group). Roentgenographic, histologic, biomechanical, bone density of all animals were examined at the end of 4th and 6th week after surgery.ResultsAt 4th week, radial bone defects healed in group A. The healing rates from group A to group E were 100%, 50%, 33%, 0%, and 0% respectively at 6th week.ConclusionRSMSCs modified by AdrhBMP-2 ex vivo in combination with DBM can repair radial longer segemental defect in rabbit. It's possible to be used in the clinical treatment of longer segemental bone defect.

Objective To observe the change of insulin-like growth factor 1 (IGF-1)before and after glucocorticoid (GC) therapy and to explore the effect of its change on bone metabolism in primary nephrotic syndrome (PNS) patients.Methods A total of 39 PNS patients with mean age of (36.73±12.15) years received GC therapy were selected from January 2008 to August 2009 in our hospital.Serum IGF-1,albumin,calcium,phosphorus,parathormone (PTH),25hydroxy vitamin D3,bone gla protein (BGP),degradation products of C-terminal telopeptides of type I collagen (CTx),24-hour urinary protein excretion and the ratio of urinary calcium to creatinine were measured at five time points-before GC therapy,4 weeks,8 weeks,12 weeks and 24 weeks after the use of GC.BMD was also detected at the same time points.Correlations among indexes were analyzed by Pearson.Results Thirty-six PNS patients fulfilled the follow-up and had complete clinical data,while other 3 patients lost.After GC treatment,serum calcium and 25hydroxy vitamin D3 were significantly increased in a time-dependent manner and were negatively correlated with 24-hour urinary protein excretion (r=-0.749,r=-0.831,P＜0.05,respectively).Serum BGP and IGF-1 were decreased after GC therapy in a time-dependent manner while CTx was significantly increased until week 12 after treatment (P＜0.05).Compared with pre-treatment,BMD of various parts had no significant difference at week 4; BMD of lumbar spine (L1-L4) was significantly decreased until week 8 (P＜0.05); BMD of femoral neck and femoral shaft was significantly decreased at week 24 (P＜0.05).IGF-1 was positively correlated with BGP and BMD (r=0.896,r=0.495,P＜0.05) and negatively correlated with serum CTx (r=-0.697,P＜0.05 ).Conclusions Serum IGF-1 level decreases in a time-dependent manner after GC treatment,which is correlated to BGP,CTx and BMD.Glucocorticoid treatment affects bone metabolism through IGF1 pathway possably in patients with PNS.IGF-1 may be used as a new bone biochemical marker of glucocoritcoid - induced osteoporosis.

Objective To study the association of bone metabolism with the degree of proteinuria in patients of chronic kidney diseases (CKD). Methods A total of 71 CKD patients diagnosed as primary glomerulopathy were randomly selected from 2008.1-2009.5 in the First People's Hospital of Shanghai. They were classified into three groups according to proteinuria:group A of 25 patients, proteinuria ＜1.0 g/24 h; group B of 16 patients, proteinuria 1.0-＜3.5 g/24 h;group C of 30 patients, proteinuria ≥ 3.5 g/24 h. Fifty-eight healthy persons were selected from our medical examination center at the same time as control. Serum albumin, calcium, phosphorus,PTH, 25 hydroxy vitamin D3, bone gla protein (BGP), degradation products of C-terminal telopeptides of type I collagen (CTx), 24-h urinary protein excretion, and the ratio of urinary calcium to creatinine (UCa/Cr) were measured. Bone mineral density (BMD) was detected by dualenergy X-ray absorptiometry. Results Compared with control group, serum levels of calcium [(2.23±0.08), (2.13±0.09), (2.04±0.06)vs (2.37±0.12)mmol/L], 25-(OH)D3 [(50.19±6.58), (47.78±6.69), (42.42±10.85) vs (56.34±8.34) nmol/L] were significantly lower and UCa/Cr was significantly higher in A, B, C groups respectively (all P＜0.05). In group B and C, BGP was lower [(18.69±7.35), (16.13±5.76) vs (22.88±6.21) μg/L] and CTx was higher [(413.59±114.93),(516.21±314.25) vs (304.53±234.15) ng/L] (all P＜0.05). BMD was lower only in group C [(1.028±0.090) vs (1.090±0.062) g/cm2, P＜0.05]. Pearson analysis showed that 24-h urinary protein excretion was negatively correlated with serum calcium and 25 hydroxy vitamin D3, and positively correlated with UCa/Cr. UCa/Cr was positively correlated with serum CTx and negatively correlated with BGP. 25-(OH) D3 was positively correlated with BGP and negatively correlated with CTx. Conclusion Bone metabolism disorder exists in CKD patients, presenting the decrease of bone formation and the increase of bone resorption, which is associated with as the degree of proteinuria, especially in patients with nephrotic syndrome.

Objective To study the change of serum insulin-like growth factor 1(IGF-1)in primary nephrotic syndrome(PNS)patients and its relationship with bone metabolism, and to investigate the clinical significance of IGF-1 in the mechanism of bone metabolic disorders in PNS patients. Methods A total of 30 PNS patients with chronic kidney disease(CKD)stage 1 and 2 were randomly selected from 2008.1 to 2009.5 in our hospital. Serum IGF-1, albumin, calcium, phosphorus, PTH,25 hydroxy vitamin D3, bone gla protein(BGP), degradation products of C-terminal telopeptides of type I collagen(CTx), 24-hour urinary protein excretion, and ratio of urinary calcium to creatinine(UCa/Cr)were measured. Healthy control group of 61 persons were randomly selected from our medical examination center at the same time. Results Serum levels of calcium, 25 hydroxy vitamin D3 and BGP were significantly lower;CTx and UCa/Cr were significantly higher in PNS patients(P<0.05)as compared to healthy control group. BMD of PNS patients was lower but without significant difference compared with healthy control group[(1.078± 0.090)g/cm2 vs(1.090±0.062)g/cm2, P>0.05]. Serum level of IGF-1 was significantly lower in PNS patients and was positively correlated with BMD and BGP,and negatively correlated with 24-hour urinary protein excretion and CTx. Conclusions Bone metabolic disorder exists in PNS patients with the appearance of decreased bone formation and increased bone absorption.Serum level of IGF-1 has good correlations with bone biochemical markers.which may be used as a new bone biochemical marker of bone metabolism in kidney disease.

To determine the fatty acid composition in the oil of Semen Momordicae to evaluate itspractical use. Fatty oil was obtained by Soxhlet extraction with petroleum ether and converted to methylester derivatives by methanolic potassium hydroxide. Contents of the resultant methyl esters were then de-termined by GC-MS. Eight fatty acids were characterized and determined. Results of the study may pro-vide some information for the exploitation and utilization in the oil of seed of Momordicae cochinchinensis(Lout.) Spreng.

Objective To detect de novo development of anti-HLA antibodies after renal transplantation, and to investigate their influence on graft function. Methods 384 kidney recipients,who were negative for anti-HLA antibody before transplantation, were monitored for anti-HLA antibodies over a period of 3-96 months, and a sensitive enzyme-linked immunosorbent assay (ELISA) was used to detect anti-HLA antibodies. HLA antibody ＞10 % was defined as positive levels. Results Among 384 recipients tested, 318 recipients (82. 8 %) were negative for anti-HLA antibody after transplantation; 66 recipients (17. 2 %) developed de novo HLA antibodies, 3 recipients with HLA class Ⅰ, 61 with HLA class Ⅱ, 2 with both HLA class Ⅰ and Ⅱ. According to amino acid residue matching, 7 cases developed de novo antibodies among 92 recipients with 0 HLA-DR mismatches,compared with 59 cases among 292 recipients with 1-2 mismatches, which showed significant difference between two groups (P＜0. 01 ). 87. 4 % (278/318) recipients negative for HLA antibodies after transplantation achieved good graft function, in comparison with 65. 2 % (43/66) recipients positive for HLA antibodies (P＜0. 05). Conclusion De novo production of HLA antibodies posttransplantation may be closely associated with HLA-DR mismatch. De novo HLA antibodies posttransplantation might damage graft function and reduce graft survival rate. The detection of de novo development of anti-HLA antibodies after renal transplantation has clinical significance for assessing renal allograft function.

Objective To investigate the expression of Notch 1 receptor in renal tissues of patients with hepatitis B virus associated-glomerulonephritis (HBV-GN) and its role in the pathogenesis of HBV-GN.Methods A total of 48 patients with HBV-GN confirmed by renal biopsy during 2008-2010 were enrolled in the study.Distribution of Notch1 receptor in renal tissue of HBV-GN was detected by immunohistochemistry and the association between the distribution of Notch1 receptor and HBsAg was examined by double-label immunofluorescence assays.Correlations of Notch1 receptor expression with renal pathology and clinical parameters of HBV-GN were analyzed.Results Notch1 receptor distributed mainly in renal tubular epithelial cells and interstitial area as brownish red granules,and a few expression in glomerulus was also found.The positive score of Notch1 receptor expression in HBV-GN patients was significantly higher as compared to primary glomerulonephritis patients with serum HBsAg positive or negative and normal renal tissue controls.Notch1 receptor expression was more obvious in membrano-proliferative glomerulonephritis (MPGN) and mesangial proliferative nephritis (MsPGN) patients,but there was no significant difference among the different pathology groups.Distribution of Notch1 receptor was consistent with the distribution of HBsAg and its intensity was positively correlated with renal interstitial fibrosis (r=0.473,P=0.001),tubular atrophy (r=0.690,P=0.000),inflammatory cell infiltration (r=0.616,P=0.000).Negative correlation was found between renal function and the intensity of Notch1 receptor (r=-0.393,P=0.006).Conclusions Notch1 receptor expression increases in the renal tissues of HBV-GN patients and distributes mainly in renal tubular epithelial cells and interstitium,which is consistent with the distribution of HBsAg.Its intensity is closely correlated with renal interstitial lesions and renal function.Abnormal expression of Notchl receptor in renal tissue of HBV-GN may be involved in the progress of HBV-GN.

ObjectiveTo investigate the expression and distribution of Toll-like receptor 4 (TLR4) in renal tissue of HBV associated nephropathy (HBV-GN) and its role in the pathogenesis and clinical manifestations of HBV-GN.MethodsRenal tissues were sampled from 48 HBV-GN patients confirmed by renal biopsy and 154 non-HBV-GN patients.The distribution of TLR4 in renal tissue and the relationship between the distribution of TLR4 and HBsAg were detected by immunohistochemistry.Integrating case record,correlations between the expression of TLR4 with clinical parameters including pathology,glomeruli,kidney tubules lesions,renal interstitial inflammatory infiltration and blood serum HBV were analyzed.ResultsTLR4 mainly distributed in the renal tubular epithelial cells and interstitial areas as brownish red and granular,which was in consistent with HBsAg distribution.The TLR4 positive rate and score in HBV-GN group were higher than those in non-HBV-GN group (P ＜ 0.05 ).TLR4 positive score was slightly higher in mesangial proliferative glomerulonephritis group and focal segmental glomerulosclerosis group,which had no significant difference (P ＞ 0.05).Kidney tubules lesions were strongly associated with TLR4 expression (r =0.748,P ＜ 0.001 ) which increased with aggravation of renal interstitial fibrosis ( r =0.569,P ＜0.001 ),tubular atrophy ( r =0.577,P ＜ 0.001 ) and inflammatory cell infiltration ( r =0.684,P ＜0.001 ).No obvious correlation with glomeruli lesions was observed ( r =0.293,P =0.053 ).Negative correlation could be seen between TLR4 and the renal function ( R2 =0.784),systolic blood pressure ( R2 =0.869),high sensitivity C-reactive protein (R2 =0.979) and urinary protein (R2 =0.615 ) by regression analysis.Other clinical parameters had no statistical significances.ConclusionsThe expression of TLR4 is abnormal in the renal tissue of HBV-GN patients,mainly in renal tubular epithelial cells and interstitial,which is consistent with the distribution of HBsAg.Its intensity is closely related with renal interstitial lesions,renal function changes and inflammatory cell infiltration.A speculation,that HBV can promote abnormal expression of TLR4 in renal tissues of HBV-GN which may be involved in the lesion progress of HBV-GN,is made upon our study.

Objective To identity whether there is muscle atrophy phenomenon in end-stage kidney disease patients and to detect the level of transcription factor Foxo1 and the activity of ubiquitin-proteasome system.Methods Twenty-two patients in chronic kidney disease (CKD) stage 5 were selected and their mean muscle cross sectional area was measured.mRNA and protein levels of Foxo1,Atrogin-1,MuRF1 in rectus abdominis biopsies obtained from consecutive patients were detected.Control biopsies were obtained from 8 healthy subjects during elective surgery for abdominal wall hernias and 6 subjects during elective surgery for adenomyosis.Results Compared with the control group,cross sectional area of muscle fibers decreased and the transcription and protein levels of Foxo1,Atrogin-1,MuRF1 were upregulated in CKD group(P＜0.05).Protein level of p-Foxo1 decreased in CKD group(P＜0.05).Conclusion There exist muscle atrophy phenomenon in CKD patients,which may associate with the upregulation of Foxo1 and activation of ubiquitin-proteasome system.

Objective To develop a superparamagnetic iron oxide nanoparticles ( SPIO ) based on MRI probe specifically targeting human epidermal growth factor receptor 2 (HER2) and explore its value as MRI positive contrast agents in vitro.Methods (1) The superparamagnetic iron oxide ( PS) was obtained by means of classical coprecipitation in polylactic acid solution , then coupled with fluorescein isothiocyanate (FITC) labeled LTVSPYW to develop the targeted probe ( FITC-LTVSPWY-PS).The particle size was measured under transmission electron microscope.Relaxation rate was detected by 3.0 T MR scanner.(2) Climbing films of human breast cancer cell MCF-7 were prepared and incubated with FITC-LTVSPWY-SPIO, then fluorescence distribution was observed under inverted microscope.And distribution of iron particles was confirmed by prussian blue staining.(3) MCF-7 and MDA-MB-231 cells were incubated with FITC-LTVSPWY-SPIO and PS, respectively.MCF-7 incubated with FITC-LTVSPWY-PS were used as experimental group, MCF-7 treated with PS as control group , and cells added with nothing as blank group.There were 3 samples in each group.The MR imaging was performed only once and T 2 WI signal intensity of cells was recorded.The comparison of T 2 signal intensity among groups was conducted by using one-way ANOVA.Results The core and surface size of nanoparticles were (13.9 ±1.6) nm and (122.0 ±5.5) nm respectively.Zeta potential and relaxation rate of the FITC-LTVSPWY-PS were ( -30.7 ±2.2 ) mV and 70.7 m· M-1 · s-1 respectively, and the PS were (28.1 ±2.8) mV and 72.1 m· M-1 · s-1 respectively.The fluorescence could be seen on the surface of MCF-7 cells, and the prussian blue staining showed that FITC-LTVSPWY-PS could specifically target HER 2-positive cells.The low signal on T 2 WI was observed in MCF-7 cells incubated with FITC-LTVSPWY-PS, whereas cells treated with PS and blank group showed equal signals , the T2 values were ( 61.8 ±5.7 ) , ( 101.6 ±2.5 ) and ( 103.5 ±1.9 ) ms respectively.Significant difference existed among these groups ( F =355.698, P <0.05 ).Conclusions The MR targeting probe FITC-LTVSPWY-PS was prepared successfully , its physical characterization and magnetic properties could target the HER 2 highly expressing on the surface of breast cancer cells and meet the need of targeted imaging.It provides an important tool for MR molecular imaging.