Automated urine analysis poses the advantage of rapid turn around time and is suitable for initial screening of large number of samples.However,standardized manual microscopic examination of urine sediment remains valuable for a variety of differential diagnosis,such as screening the potential causes of hematuria,to predict proliferative and non-proliferative renal pathological injury. Urinary sediment scoring system is often used to differentiate acute tubular necrosis (ATN) from pre-renal acute kidney injury (AKI).The urinary podocytes serves as a marker of glomerular injury to determine the location of and to monitor activity of glomerular lesions,as well as to differentiate focal segmental glomerulosclerosis (FSGS) and minimal change disease (MCD).These useful analyses shows the importance of urine microscopy in the clinical practice of nephrology.( Chin J Lab Med,2012,35:780-783)

Macrophages have been identified to play an important role in acute and chronic renal injury through exerting multiple biological effects.This review focus on the origin of macrophages in kidney, mechanism of renal injury and strategy of inhibiting macrophage infiltration. [

Objective To investigate the feasibility whether clearance of iohexol can be a reliable, sensitive and safe method for the determination of GFR. Methods The GFR of 19 patients with different renal functions were examined using clearance of 99m Tc-DTPA and clearance of iohexol. Then the correlation of them was analyzed. Serum and urinary iohexol was determined by X-ray fluorescence analysis. Results, These two methods were significantly correlated (r = 0. 98). Conclusion Clearance of iohexol is a safe, comfident, no-radioactivity method for the clinical practice of GFR determination.

Objective: To investigate whether hypoxia can affect the expression and secretion of connective tissue growth factor(CTGF) and fibronectin(FN) in primary cultured rat renal cortical myofibroblasts . Methods: The primary cultured rat renal cortical myofibroblasts were subjected to hypoxic (1%O_2) or normoxic (21% O_2) conditions for a variety of times. The protein levels of HIF - 1?, CTGF and FN protein were analyzed by Western blotting in both the whole cell lysates and supernatant culture medium 6 h,12 h and 24 h after incubation, respectively. RT-PCR was carried out to measure the levels of FN mRNA at different time points (2 h,3 h,6 h and 12 h). The activity of gelatinase MMP-2 and MMP-9 in the supernatant from the cultured cell medium was assayed by gelatin zymography. Results: The expression of HIF - 1?was induced at h6 in cells under hypoxia incubation. The levels of cellular CTGF protein were increased in hypoxia treated myofibroblasts at h6 (175%?52%),significantly elevated at h12 (347%?67%,P

AIM: To investigate the effect of hypoxia on the myofibroblast transdifferentiation from fibroblasts,and associated signaling of hypoxia on the production of collagen Ⅰ in cultured rat renal cortical myofibroblasts.METHODS: The study is composed of two relevant parts.In the first part,a normal rat renal interstitial fibroblast cell line NRK-49F was treated with hypoxia(1% O2) or normoxia(21% O2) for 6 h,12 h and 24 h.The expression of hypoxia inducible factor-1?(HIF-1?) was examined by Western blotting in order to make sure the hypoxic condition is reliable.The myofibroblast transformation from fibroblasts induced by hypoxia was assayed by detecting the protein levels of ?-smooth muscle actin(?-SMA).In the second part,the object was done on the primary cultured rat renal cortical myofibroblasts.Myofibroblasts were subjected to hypoxic or normoxic conditions for variety of times.The levels of HIF-1? in cell lysates and collagen I protein in supernatant culture medium and the activation of extracellular signal-regulated kinase(ERK)1/2 MAPK pathway were analyzed by Western blotting.RT-PCR was carried out to measure the levels of collagen I mRNA at different time points(2 h,4 h and 6 h).The distribution of HIF-1? in myofibroblasts was demonstrated by immunocytochemistry.The changes of collagen I production were detected after PD98059,a specific inhibitor of ERK1/2 activation pretreatment and during the hypoxia incubation.The activity of gelatinase matrix metalloproteinase-2(MMP-2) and MMP-9 in the supernatant medium from the cultured cells were assayed by gelatin zymography.RESULTS: Significant increased levels of HIF-1? protein appeared in cell lysates under hypoxia for 6 h.Furthermore,HIF-1? was translocated into nuclei of myofibroblasts after 6 h exposure of myofibroblasts to hypoxia.The levels of ?-SMA protein increased in NRK-49F under hypoxia for 12 h(187%?32%,P

AIM A&A treated PAN rats, and to observe the effects of A&A on MAPK signaling pathway. METHODS Rats were divided into control, PAN, A&A treated PAN (A&A) and enapril treated PAN (ACEI) groups. The pathological lesion was observed under a light microscope. Immunohistochemistry combined with semi quantitive method was used to investigate the following parameters: cell number, ? SMA expression and extracellular matrix deposition. Expression and phosphorylation of protein kinases ERK, JNK and p38 were assayed. RESULTS In PAN rats, A&A suppressed ? SMA expression, which was closely correlated to cell proliferation, and extracellular matrix accumulation in glomerular mesangium. A&A significantly attenuated ? SMA expression in the tubulo interstitial area which was also parallel to the renal interstitial fibrosis.In this study, expression of all subtypes of MAPK had no difference between control and PAN groups. Compared with the inactivation of ERK and p38, phosphorylation of JNK was observed in glomeruli, renal tubules and interstitial cells in PAN rats, which was also inhibited by A&A treatment. CONCLUSION The inhibitory effect of A&A on phenotypic changes of renal resident cells, especially glomerular mesangial cell, may participate in its renal protective mechanisms. This effect, at least partially, was mediated by down regulated JNK activation.

Objective:To observe the expression of connective tissue growth factor(CTGF) and its receptor-low density lipoprotein receptor-related protein (LRP), and the relevant signaling pathway for the regulation by long-term high glucose exposure in cultured podocytes. Methods:The effects of high glucose on the expression of CTGF and its receptor LRP were analyzed by western blotting. The activation of mitogen activated protein kinase ( MAPKS )signaling pathway by high glucose was also examined. Results: Basal levels of CTGF were observed in cultured mouse podocytes, the levels of CTGF protein were increased by high glucose medium groups on the 2nd day, reached the peak on the 4th day(P0.05).The levels of CTGF expression in normal glucose and mannitol glucose groups did not change markly. High glucose medium induced phosphorylation of ERK_ 1/2 at as early as minute 30, reached the peak at hour 6; maintained the activity at hours 12 and 24, and declined to the basal level at hour 48. However, phosphorylation of ERK_ 1/2 was not detected in normal glucose and mannitol glucose groups. Blockade of phosphorylation of ERK_ 1/2 with PD98059, a specific ERK_ 1/2 activation inhibitior, did decrease the high glucose-triggered expression of CTGF protein in 4 days. High glucose had no effect on the expression of LRP protein at each time point. Conclusion: Acute high glucose (2-4 days)stimulated the expression of CTGF protein via ERK_ 1/2-dependent signaling pathway in cultured podocytes, while cultured in high glucose for 6-8 days, the podocytes did not increase its CTGF level. Long-term high glucose had no effect on the expression of LRP in podocytes.

Objective To study the effect of tanshinone ⅡA on the expression of P-selectin and ICAM-1 after cerebral ischemia reperfusion (I/R)injury in rats. Methods Rats were randomly divided into 4 groups: Sham operated group, I/R group, low dose Tan ⅡA treated group and high dose Tan ⅡA treated group. The focal middle cerebral artery occlusion (MCAO) model was made by suture-occluded method. Rats were pretreated with Tan ⅡA, ig for 3d,respectively before MCAO. After 90min MCAO following 24 hours of reperfusion, the expression of P-selectin and ICAM-1 was detected with using immunohistochemistry method. Result Compared with sham operated group, the expression of P-selectin and ICAM-1 increased after reperfusion for 24 hours in the ischemic territory(all P＜0.01).Compared with I/R group, the expression of P-selectin and ICAM-1 decreased in a dose dependent manner in low and high dose Tan ⅡA treated group(P＜0.01).Compared with that of I/R group, cerebral infarction volume was decreased in a dose dependent manner in low dose Tan ⅡA treated group and high dose Tan ⅡA treated group(all P＜0.01).The change of ischemic impairment in low or high dose Tan ⅡA treated group was less than that in IR group, and the change of ischemic impairment in high dose Tan ⅡA treated group was less than that in low dose Tan ⅡA treated group. Conclusion Tan ⅡA may reduce cerebral ischemia-reperfusion inflammation injure by decreasing the expression of p-selectin and ICAM-1.Tan ⅡA plays protective effect on cerebral ischemia injury, especially when high dose of Tan ⅡA(30mg/kg)was used.

AIM: To investigates the role of phosphatidic acid (PA) in the expression of several inflammatory mediators produced by glomerural macrophages (GM?). METHODS: The study was performed on a rat model of accelerated anti-glomerural basement membrane (anti-GBM) glomerulonephritis (GN). GN-GM? were isolated and identified. Peritoneal M? (P-M?) of both normal and GN rats were used as controls. Block and reverse test were investigated with rhIL-1? stimulated, lisofylline (LSF) and phosphatidic acid (PA). Macrophage expression of ICAM-1 and TGF-? 1 were assessed at the level of protein and gene by immunocytochemistry, northern blot and RT-PCR. RESULTS: (1) After stimulated with rhIL-1?, GN-GM? produced much more ICAM-1, MCP-1 and TGF-? 1 than P-M?, and it's gene expression was similar as protein product. (2) mRNA expression of these factors was up-regulated again after the GN-GM? were pretreated with LSF then PA was added. CONCLUSIONS: Since GN-GM? plays an important role for PA in the mediation of glomerular injury, inhibiting of PA production is the keypoint of blocking M? mediated inflammatory effects. LSF may be an effective medicine in therapy for acute inflammatory forms of GN.

AIM: To compare the renoprotective effects of angiotensinⅡtypeⅠreceptor antagonist (AT 1RA) with that of angiotensin converting enzyme inhibitor (ACEI) and to investigate their influences on intrarenal renin-angiotensin system. METHODS: Experimental nephrotic syndrome model was induced in SD rats with repeated peritoneal injections of puromycin. Twenty-eight rats were randomly divided into four groups: normal control, nephrotic control, ACEI-treated and AT 1RA-treated group. Serum, urine and renal tissue were collected for study 12 weeks later. RESULTS: The urine protein was less and renal function was better in both treated groups. The glomerular and interstitial injury indexes of both ACEI and AT 1RA treated rats were lower than that of nephrotic control rats and had no significant difference between the two treated groups. The renal local ACE activity and angiotensinⅡ of nephrotic control group were significantly higher than that of normal control group and the two treated group(P