Objective: To screen the changes of immune-associate genes expression which is related with the ageing using cDNA microar-ray.Methods:The mRNA from the spleens of young and aged rats were extracted respectively and reversely transcribed to cDNAs with the incorporation of different fluorescent-labeled dUTP as the hybridization probes. The mixed probes were hybridized to the cDNA microarray that contains the cDNA products of 416 immune-associated genes. After high-stringent washing, the cDNA microarray was scanned for the fluorescent signals and showed the gene expression differences between the young and the aged. Some biochemical assays were used to confirm the physiological differences between the young rats and the aged rats. Results: Among the examined genes, 13 down-regulated genes were identified. These genes correlated with immuned response, cell proliferation, cell differentiation and DNA/RNA repair. Only one gene which encoded ?-amylase was much higher in the aged than that in the young. Conclusion: Further analysis of the differenially expressed immune-associated genes based on cDNA microarray will be helpful for understanding the molecular mechanism of the ageing.

Objective To observe P27 expression and proliferation of mesangial cells of rats with Thy1 glomerulonephritis and to evaluate the interfering effect of valsartan, a specific angiotensin Ⅱ receptor antagonist, on Thy1 glomerulonephritis. Methods Rats were divided into normal control group, Thy1 glomerulonephritis group and valsartan treatment group. Following the onset of glomerulonephritis, renal morphological changes were observed on days 1, 3, 5 and 7, and proliferating cell nuclear antigen(PCNA) and P27 expression in glomerulus were measured by immunohistochemistry and Western blotting. Results High expression of P27 was found in quiescent mesangial cells of normal rats, but decreased expression in mesangial cells of rats with Thy1 glomerulonephritis, accompanying with the proliferation of mesangial cells. In valsartan treatment group, glomerular hypercellularity, mesangial matrix expansion and PCNA expression were significantly reduced as compared with those of the untreated rats with Thy1 glomerulonephritis from day 3 to day 7( P

Objective To evaluate the soothing effects of hemodiafiltration on patients with refractoriness nephrotic edema and its influence on prognosis of nephrotic syndrome.Methods All the 15 cases involved had undergone hemodiafiltration periodically during acme edema and administered with the same standard loading dose of prednisone for 8 weeks.Meanwhile,clinical and lab indexes (urine volume,Cr,Bun,urinary protein,albumin) were measured.Results All patients improved in urine volume and renal function gradually after the treatment of hemodiafiltration for 1 to 3 times.By the 8th week of post-hemodiafiltration,urinary protein 24 h declines obviously[(6.42?2.31)g/d to (0.87?1.24)g/d].Conclusion Hemodiafiltration is an effective treatment in relieving retention of water,increasing the remission rate of protein urine,recovering patients' renal function and improving their prognosis to nephritic syndrome patients with severe refractory edema.

AIM: To express the synthesized human insul in like growth factor I (hIGF-1) gene in E.coli with high expression level a nd explore the way to increase the efficiency of factor Xa cleavage. METHODS: The gene of hIGF-1 was designed and synthesized accordi n g to the preference of E.coli. A fusion protein with a recognized site of fa ctor Xa between CBD (cellulose binding domain) and hIGF-1 was expressed and puri fied by cellulose affinity chromatography. MTT method was used to assay the bioa ctivity of CBD-IGF fusion protein. hIGF-1 was released by factor Xa. In order to improve the sensitivity of fusion protein to factor Xa, the short flexible pept ide (Gly-Thr-Gly- Gly-Gly-Ser-Gly) was added before the recognized site of fac tor Xa. RESULTS: SDS-PAGE results indicated that the CBD-IGF fusion prot ein was expressed and purified . Biological assay results indicated CBD-IGF fusi on protein could promote the growth of NIH3T3 cell. The short flexible peptide (Gly-Thr-Gly-Gly-Gly-Ser-Gly), which was added before the recognized site of f actor Xa, improved the sensitivity of fusion protein to factor Xa. CONCLUSION: CBD-IGF fusion protein with bioactivite are expresse d and purified. The amio acid sequences changes between the site recognize of fa ctor Xa can help to improve the cleavage efficiency of Factor Xa.

AIM: To construct engineered E.coli strains which can express nattokinase with fibrinolysis activity using gene engineering technology. METHODS: The pro-nattokinase (pro-NK) gene was amplified by PCR and inserted into expression vector pET3c. The recombined plasmid pENK which expressed the fusion protein of pro-NK and 22 amino acid peptide was then transferred into lysogenic host strains BL21(DE3)pLysS - and BL21(DE3)pLysS +. Both SDS-PAGE and the fibrin plate assay were used to examine the expression and the activity of the target protein. RESULTS: SDS-PAGE assay showed the fused gene encoding 42 kD fusion protein was expressed in both expression strains pENK-(DE3)pLysS - and pENK-(DE3)pLysS +, and the fibrin plate assay indicated that the expression product had fibrinolysis activity. pENK-(DE3)pLysS - exhibited the basal expression of the target gene, while fusion protein was only induced by IPTG in pENK-(DE3)pLysS +. Basal expression of the fused toxic gene in pENK-(DE3)pLysS - led to bacteriolysis and hollow lawns. CONCLUSION: A pro-NK fusion protein with fibrinolysis activity is successfully expressed in E.coli , which lay a foundation for the exploitation of nattokinase.

Objective To investigate the therapeutic effect of endothelial progenitor cells(EPCs) in ameliorating rat progressive focal segmental glomerular sclerosis(FSGS) model induced by adriamycin.Methods Bone marrow mononuclear cells from male SD rats,after cultured by adherence method,were identified as EPCs.Female SD rats were divided into normal control group,adriamycin induced renal disease(ADR) group,EPCs transplantation group.ADR group and EPCs group underwent unilateral nephrectomy and received 5,3 mg/kg of adriamycin via tail vein 1 week and 2 weeks after operation,while the control group underwent sham operation and received 0.9% sodium chloride solution of equal volume.The whole body irradiation by 5 Gy X ray was done 1 week after the 2nd injection of adriamycin,then immediately 1?106 EPCs were transplanted via tail vein.The rats in control group and ADR group were only injected with 0.9% sodium chloride solution after whole body irradiation.The body weight and urine protein were measured before operation(0 week) and 4(1 week after EPCs transplantation),8,12 and 16 weeks after nephrectomy.Y chromatosome incorporation was detected with in situ hybridization at the 4th and 16th week.The histological and ultrastructural changes of kidney were evaluated at the 16th week.Results At the 4th and 16th weeks,Y chromatosome positive cells could be found incorporation in the area of glomerular and tubular epithelial cells.Since the 4th week,the weight of rats in both ADR group and EPC group became significantly less than that in control group and since the 8th week that in ADR group became less than that in EPC group(P

ObjectiveTo investigate the treatment and safety of interferon ? plus Ribovirin for chronic hepatitis C after kidney transplantation. MethodsFive patients with chronic hepatitis C after kidney transplantation were administered with interferon ? (50 ?g) subcutaneously once a week, plus Ribovirin (600 mg) orally once daily. The levels of HCV-RNA, ALT and serum creatinine in patients’ serum were monitored monthly. ResultsFour in 5 patients presented normal ALT and negative HCV-RNA in serum 12 weeks after treatment, and obtained sustained viral response 24 weeks after interferon ? plus Ribovirin therapy. During treatment, renal graft rejection did not occur. The most frequent side-effects were the decrease of leukocyte and hemoglobin, myalgia and fever, but did not influence the course of treatment. ConclusionCombination of interferon ? with Ribovirin can be a valid therapeutic option in renal transplant recipients with hepatitis C, and shows no influence on the renal function.

ObjectiveTo explore the clinical and pathological characteristics in diagnosing the renal diseases with diffuse glomerular nodular changes. MethodsThe clinical and pathological data of 127 patients whose renal pathological feature was mainly diffuse glomerular nodular changes were retrospectively analyzed. ResultsThe most common diagnosis was diabetic nephropathy and renal amyloidosis, and sometimes light chain deposit diseases and Mix cryoglobulinemia. ConclusionDiffuse glomerular nodular changes occurred in many diseases. We should make differential diagnosis based on the clinical symptoms, the results of laboratory examinations and the renal pathological characteristics.

Objective To observe the impact of transplant nephrectomy on panel reactive antibodies and a secondary renal transplantation.Methods Panel reactive antibodies in 15 patients with a failed renal transplant admitted in our hospital between 2004 to 2007 were measured before transplantation,before and 1 month,6 months,12 months after transplant nephrectomy,and the pathological changes were observed after transplant nephrectomy.Results Panel reactive antibodies was increasing after renal transplantation,and reached the highest level one month after transplant nephrectomy,then grandually got down.New HLA allosensitization sites were discovered after transplant nephrectomy.Large amount of C4d was stained in failed transplant.Conclusion Serum PRA increased after transplant nephrectomy.New HLA allosensitization sites were found,which may be useful in HLA matching.