Alterations in the balance between synthesis and degradation of extracellular matrix (ECM) and its remodeling may result in an accumulation of ECM molecules and lead to glomerulosclerosis. Recent studies have focused on the role of degradative systems, especially the roles of plasminogen activators/plasminogen activator inhibitors (PA/PAI), matrix metalloproteinases, and their inhibitors[GK2*4/5!2*4/5] (MMP/TIMP) in the initiation and pathogenesis of renal diseases. Previously, attention has been paid to the study of inhibitors of ECM degrading enzymes, such as PAI and TIMP. Recent researchs showed that there existed complex dynamic expressions of enzymes and their inhibitors. Although many studies have tried to elucidate the pathogenesis of renal diseases, the exact underlying mechanisms are still not completely understood. For better understanding of the mechanism of chronic progressive renal diseases, the underlying genetic and molecular regulation of each component of PA/PAI and MMP/TIMP systems should be elucidated in different renal physiological and pathophysiological processes. Future studies are needed to manipulate activity or expression of these proteinases in order to treat and/or prevent glomerular diseases.

The prevention and control of chronic kidney disease(CKD)and end-stage renal disease(ESRD)has become an important public health problem.This article has outlined the status,hazards,and problems in prevention of CKD and ESRD,briefly described the work having been done by the Chinese Society of Nephrology of the Chinese Medical Association,and proposed countermeasures for future prevention and treatment of ESRD,aiming at improving knowledge of the urgency of combating ESRD by the society,government,public,and medical staff in order to improve the prevention and treatment of ESRD in China.

Objective To introduce the current development of nephrology during the period of "Eleventh-Five-Year Plan" in PLA medical circles,to serve as a reference for the further development of nephrology in PLA.Methods Literature concerning nephrology published domestically and abroad in past 5years were retrieved,and the progresses,achieved domestically and abroad,especially in PLA,on new concept,diagnoses and therapy of common nephropathy,and clinical applications of new drugs and techniques were emphatically analyzed.Results Great progresses have been made during the period of "Eleventh-Five-Year Plan" on basic researches,clinical applications and substitution therapy in nephrology,and outstanding achievements have been acquired on basic and clinical researches of chronic nephropathy and acute renal injury,drug treatment of renal diseases and continuous renal replacement therapy.The PLA medical personnel participated in the formulation of "Diagnostic and Therapeutic Standard of Renal Diseases" ,furthered the academic exchanges between the military and civilian circles,both domestically and abroad.The academic level of PLA in nephrology was raised markedly with obvious features and preponderance.Conclusion During the period of "Twelfth-Five-Year Plan" ,the focus of nephropathy researches should be put on the enhancement of the ability of military health service,integrated control and comprehensive remedy of acute renal injury induced by combat wound,trauma and stress injuries.It is important to stably retain the superiority on basic and clinical researches of chronic nephropathy,acquire more achievements,make greater contributions to raising the professional level of diagnosis and treatment of kidney diseases.

Chronic kidney disease(CKD)is increasingly recognized as a global public health problem.Uncontrolled complications of CKD,especially cardiovascular diseases,contribute greatly to the premature death and unfavorable prognosis.Recent evidence shows that CKD complications may occur earlier than previously thought.CKD complications deserve early detection and active treatment.Periodical follow-up and regular check should be done to adjust the therapeutic condition.Clinical practice guideline or recommendation based on evidence-based medicine is essential for management of CKD complications.Personalized treatment should be considered to improve survival and quality of life,and to make patient return to society.

Animal model of aged rat nephrotoxicity was induced by i. p. administrationof gentamycin in a dose of 140mg/kg/day. Part of those rats were treated with CordycepsSinensis(CS), Epimedium(Ep) and Astragalus Membranaccus (AM) in form of decoc-tion per Os and others seryed as control. The results were summarized as. 1. The nephro-toxicity of gentamycin was aggrevated with age. CS, Ep and AM are effective drugs inpreventing the tdeular damage caused by gentamycin in rats. The pathological changes ofrenal tuoules of the rat groups which treated with CS, Ep and AM were less severe thanthat of the control. 2. CS, Ep and AM could prevent the decline of renal cortical Na~+-K~+-ATPase activity of aged rat induced by gentamycin.

Objective To investigate the clinico-pathological characters of acute and chronic Aristolochic acid nephropathy, and analysis the pathological mechanism of chronic Aristolochic acid nephropathy.Methods 26 cases of aristolochic acid nephropathy diagnosed in our department were examined. They were divided into acute and chronic group by their pathological characters. Immunohistochemical staining for the expression of collagen III, PAI-1, TIMP-1 and PCNA was done in renal biopsy specimens.Results There were 11 acute cases and 15 chronic cases. Compared with acute cases, there were more female, longer duration of the medicine intake[ (142.3?52.7 months of chronic cases and 4.5 ? 2.7 months of acute cases), higher degree of hypertension[(156.7?32.4) mm Hg of chronic cases and 127.3?24.2 mm Hg of acute cases], 24 hour urinary protein,anemia, glomerulosclerosis, tubular atrophy, interstitial fibrosis and artery lesions in chronic patients(Pall

Objective To prepare the anti-hUART1 polyclonal antibody and investigate the subcellular localization of hURAT1 protein. Methods The full-length hURAT1 gene was obtained by RT-PCR and inserted into the fusion expression vector pEGFP-N3,the predicated antigen epitope was cloned into GST fusion protein expression plasmid pGEX-5X-1, transformed into E. coli BL21 cells for expressing recombinant GST-hURAT1 protein induced by IPTG. The purified hURAT1-GST fusion protein was employed to immunize rabbit for preparing the polyclonal antibody. The expression of hURAT1 was analyzed by western-blot and immunohistochemistry in human kidney. pEGFP-hURAT1 was transfected into the LLC-PK1 cell in order to observe the subcellular localization of the gene using confocal microscopy. Results Specific anti-hURAT1 rabbit polyclonal antibody was obtained, and both Western-blot and immunohistochemistry showed that hURAT1 was expressed in the human kidney brush border,localized in the apical membrane of the LLC-PK1 cell. Conclusions hURAT1 protein was a membrane protein located in renal proximal tubule, which could be detected in the apical membrane. The anti-hURAT1 polyclonal antibody could be used for studying the physiological function of hURAT1 and its pathology.

Objective To investigate the effects of shRNA produced by PCR on the suppression of the expression of exo- or endogenous genes. Methods The specific primers were designed, with which the shGFP-RNA and shFN-RNA were produced by PCR. The shGFP-RNA was transfected into 293T cell lines together with pEGFP plasmid, then the cells were detected by laser confocal microscopy 48h later. The shFN-RNA was transfected into rat mesenteric cell lines, then the cells were collected 48h later and the total RNA was extracted, which was reversely transcripted to cDNA. Then the expression level of FN mRNA was examined with real-time PCR, and the expression level of FN protein was examined with Western blot analysis. Results The results of laser confocal microscopy indicated that the EGFP could be successfully suppressed by shGFP-RNA produced by PCR; the results of real-time PCR and Western blot analysis revealed that FN expression level of the cells transfected with shFN-RNA was down regulated, and the level was much lower than those tansfected with independent shRNA (P

Objective To thoroughly explore the pathophysiological roles of TIMP-1 during the progressive course of renal diseases, the study was aimed at identifying the functional phenotype of endogenous and exogenous genes in kidneys of human TIMP-1 transgenic mice. Methods Renal histological changes between 3-month-old wild type mice (n=8) and 3-month-old transgenic mice (n=8) were analyzed through PAS staining of paraffin sections. The mRNA and protein expressions of h/mTIMP-1, mTIMP-1, TIMP-2, TIMP-3, MMP-9, MMP-2, and COLⅣ?5 mRNA were detected by Northern blot and Western blot. The activities of gelatinases and TIMP-1 were examined by gelatin zymography and reverse zymography, respectively. Results No difference in histological picture in kidneys was found between wild type and transgenic mice. In contrast with wild type mice, it was found that in kidneys of transgenic mice, the mRNA and protein expressions of h/mTIMP-1 and its activity were up-regulated (P0.05). Conclusion The transgene was expressed steadily in kidneys of human TIMP-1 transgenic mice, and it induced the compensation of MMPs/TIMPs.

Objective To prepare mouse polyclonal antibody against human Nanog by genetic immunization and to identify this antibody by Western blot and immunofluorescence. Method The antigenicity fragment (A16-V101) of human Nanog (hNanog) was chosen by analysis of Accelrys software, and its cDNA (258bp) was amplified from plasmid containing full-length cDNA of hNanog, then it was cloned into pBQAP-TT to construct recombinant plasmid pBQAP-TT-hNanog for genetic immunization. Mice were immunized with this recombinant plasmid and two other adjuvant plasmids-pCMVi-GMCSF and pCMVi-FIT3L, which help to enhance the antibody's generation. After 12 weeks, we obtained mouse anti-hNanog antibody from mice blood serum. The antibody titer was determined by enzyme-linked immunoadsordent assay (ELISA), and its specificity was identified by Western blot in human renal protein. Using this antibody, we detected hNanog expression in HKC cells of hNanog-AAV2 transfection. Results Recombinant plasmid pBQAP-TT-hNanog for genetic immunization was confirmed to be correct by restriction digestion and sequencing. The result of ELISA showed that the antibody titer was 1∶3 200. This antibody recognized a band of 34kD hNanog protein in human renal protein by Western blot. Immunofluorescence showed that Nanog protein was mainly located in the nuclei in hNanog transgene HKC cells. Conclusion Genetic immunization can offer mouse anti-hNanog polyclonal antibody of high titer and high specificity.