1.Remission of severe nephrotic edema by hemodiafiltration and its effect on the prognosis of nephropathy
Rongjie YU ; Xiongfei WU ; Hongwen ZHAO
Chinese Journal of Practical Internal Medicine 2001;0(09):-
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.
2.Study of immune-associated genes in the aged rats by cDNA microarray
Rongjie YU ; Hanlin PU ; Jian LIN
Chinese Journal of Immunology 2000;0(08):-
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.
3.Effect of valsartan on P27 expression in mesangial cells of Thy1 glomerulonephritis rats
Rongjie YU ; Huibiao YANG ; Jinghong ZHAO ;
Journal of Third Military Medical University 2003;0(22):-
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
4.Synthesization, expression, purification and activity assay of hIGF-1
Yuan GAO ; Rongjie YU ; An HONG ; Zhiyin LI
Chinese Journal of Pathophysiology 1986;0(02):-
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.
5.Treatment of hepatitis C after kidney transplantation with combined interferon-? and Ribovirin
Xiaopeng TANG ; Hongwen ZHAO ; Rongjie YU ; Xiongfei WU
Journal of Third Military Medical University 1984;0(02):-
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.
6.Differential diagnosis of diffuse glomerular nodular changes
Lian LI ; Rongjie YU ; Hongwen ZHAO ; Xiongfei WU
Journal of Third Military Medical University 1984;0(02):-
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.
7.Impact of transplant nephrectomy on panel reactive antibodies
Hong LIU ; Lian LI ; Rongjie YU ; Hongwen ZHAO ; Xiongfei WU
Journal of Third Military Medical University 2003;0(10):-
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.
9.Preparation and purification of advanced oxidation protein products
Mei TIAN ; Weijing YI ; Rongjie YU ; Xiongfei WU
Chinese Journal of Tissue Engineering Research 2009;13(28):5544-5548
BACKGROUND: Advanced oxidation protein products (AOPPs) are a crucial pathogenic link to such long-term uremic complications in hemodialysis patients as immune system dysregulation, accelerated atherosclerosis, dialysis-related amyloldosis and so on. However, basic studies on AOPPs are relatively few, and one of the main reasons is the fact that it is difficult to obtain AOPPs with high pudty and biological activity.OBJECTIVE: To prepare, pudfy and indentify AOPPs, with the hope of searching for a method of preparing AOPPs with high purity and biological activity.DESIGN, TIME AND SETTING: A single sample observation was completed in the Clinical Biochemistry Section of Ecsomatics Department, Third Military Medical University of Chinese PLA from September to November in 2008. MATERIALS: Human serum albumin (HSA) was provided by Chengdu Rongsheng Company Ltd. Hitrap 26/60 sephacryl S-300 was purchased from GE Healthcare.METHODS: Hypochloric acid was used in the oxidation of purified HSA to prepare in vitro the AOPPs-modified HSA (AOPPs-HSA), which was then isolated by Hitrap 26/60 sephacryl S-300. Relative molecular mass was determined by sodium dodecyl sulfate polyacrylamide gel electropheresis (SDS-PAGE), native polyacrylamide gel electrophoresis (PAGE) and molecular weight standards. Structural features and biological activities were identified in the experiment of tumor necrosis factor α (TNF-α) release from monocytes.MAIN OUTCOME MEASURES: ①The purification and gel electrophoresis results of HSA. ②The purification and gel electrophoresis results of AOPPs. ③The dose-effect relationship between AOPPs-HSA and TNF-α release from monocytes. RESULTS: The relative molecular mass of AOPPs-HSA was 670 000 according to SDS-PAGE, native PAGE and molecular weight standards. Moreover, AOPPs-HSA could encourage the release of TNF-α from monocytes. The time effects showed that TNF-α release volume significantly increased after 6 hours of stimulation by AOPPs-HSA (1 g/L) and reached a peak at hour 12. CONCLUSION: Highly purified and bioactive AOPPs can be successfully prepared by the above-mentioned method, which builds a basis for further study on AOPPs.
10.Molecular cloning, fusion expression and bioactivity of pro-nattokinase gene
Rongjie YU ; Qiuling XIE ; An HONG ; Ju WANG ; Fenyong SUN
Chinese Journal of Pathophysiology 1989;0(06):-
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.