Objective To investigate the role of LPL in enhancing VLDL uptake in mesangial cells and modulating VLDL-mesangial interaction. Methods Human wild type LPL (LPLwt), catalytically inactive LPL (LPL194) or control alkaline phosphatase (AP) were expressed in human mesangiai cell line (HMCL) via adenoviral vectors. The expression of LPL mRNA and protein was detected by RT-PCR and immunoehemistry staining, respectively. LPL activity was assayed by radioisotope labeled liposome substrate. Cellular lipid deposition was visualized by oil red O staining and analyzed quantitatively by standard enzymatic procedures. Effect of LPL on HMCL proliferation was evaluated by colorimetric assay using MTr. MCP-1 mRNA and protein levels in treated HMCLs were determined by real-time quantitative BT-PCB and enzyme-linked immunosorbent assay respectively. For adhesion study, HMCLs were treated with VLDL for six hours, followed by one-hour incubation with Tamm-Horsfall protein-1 (THP-1) cells. Results Compared with HMCLs transfected by Ad-AP, the lever of cellular triglyceride content was sharply increased in Ad-LPLwt Wansfected HMCLs [(109.11±5.01) mg/g protein vs (23.98±3.23) mg/g protein,P<0.01] and was slightly increased in Ad-LPL194 transfected HMCLs [(36.33±2.64) mg/g protein vs (23.98±3.23) mg/g protein, P<0.05]. LPLwt amplified VLDL-driven mesangial cells proliferation. Compared to the HMCL-Ad-AP, MCP-1 mRNA and protein expression increasd by 39% (P<0.05) and 171% (P<0.01) in HMCL-Ad-LPLwt, and the amount of THP-1 cells adhering to HMCL-Ad-LPLwt was increased by 1.69-fold (P<0.05), without significant difference between HMCL-Ad-LPLI94 and HMCL-Ad-AP. Conclusions Overexpression of either active or inactive LPL in HMCLs accelerates VLDL-induced triglyceride accumulation, and enzymolysis action of LPL may be the major factor in this process. Active LPL significantly amplifies VLDL-induced proliferative effect on mesangial cells and enhances monocyte adhesion to mesangial cells through up-regulation of MCP-1. Hence, LPL may be an important contribution to initiation and progression of renal injury mediated by triglyceride-rich lipoproteins.

Objectives To study the relationship of atypical membranous nephropathy (AMN) with idiopathic membranous nephropathy (IMN) and lupus membranous nephropathy (LMN), and to explore the predictive clinical and pathological features for LMN diagnosis. Methods The patients undergone renal biopsy in PUMCH between 2003 and 2006 were selected, and were divided into group AMN (n=28), IMN (n=100) and LMN (n=45). Clinical manifestations and pathological features were compared among three groups retrospectively. The intensity of glomerular IgG subclasses was analyzed by immunohistoehemical staining among three groups semi- quantitatively. The spatial arrangement of IgG and C3 deposits was investigated by immunofluorescenee double staining among three groups by eonfocal laser scanning microscopy. Results (1) The onset age of AMN was (38±17) years and female/male ratio(F/M) was 2.5:1 in group LMN and IMN. The onset age was significantly different among three groups (P<0.01), and F/M ratio was significantly different between AMN and IMN (P=0.017). (2) The incidence of most extra-renal manifestations was less than 20% in AMN except for hematological disorder (21.4%) and serum anti-SSA antibody positivity(40.7%). (3) The incidence of subendothelial electron densedeposits in either LMN or AMN was significantly higher than that in IMN (P<0.01). (4) The percentage of IgG3 predominance in AMN and LMN giomeruli was 78.9% and 73.9%, respectively, while the percentage of IgC,-4 predominance in IMN was 61.1%. The difference was significant(IMN vs AMN and LMN, P<0.01). (5) IMN had an overlapping distribution of IgG and C3 in subepithelial deposition, which was rarely found in AMN or LMN. (6) Among the indexes differentiating LMN and IMN, the high sensitive one was non-IgC,4 predominance in glomeruli (91.3%), while the high specific ones included subendothelial electron dense deposits (100.0%), serum anti-SSA antibody (95.5%), glomendar IgG3 predominance (94.4%). Conclusions AMN with serum ANA positivity is similar to LMN in respect to pathological features and glomerular IgG subclasses, although it has few extra-renal clinical manifestations. It may represent a latent subgroup of lupus nephritis.

Objective To study the expression of lipoprotein lipase(LPL) in human glomerular mesangial cells and the effect of very low-density lipoprotein(VLDL) on the expression of LPL.Methods LPL mRNA expression,protein synthesis and activity were detected in human glomerular mesangial cells by RT-PCR,Western blot and a radio-chemical analysis respectively.Effect of VLDL on the expression of LPL in mesangial cells was detected by Western blot.Results In human glomerular mesangial cells,a 276 bp band,that was specific for human LPL,was identified by RT-PCR,and the same of a 55 kd band,specific for human LPL by western blot.LPL activity of mesangial cells was also detected in the medium after release by heparin.VLDL stimulated LPL protein synthesis in mesangial cells in a time-and dose-dependent manner.Conclusion LPL is expressed by human mesangial cells and it has catalytic activity.Expression of LPL in mesangial cells is regulated by VLDL.

Objective To evaluate the effects of propofol on apoptosis in the hippocampal neurons of fetal rats in vitro.Methods The isolated hippocampal neurons were seeded into 96-well plates or 24-well plates at a density of 5 × 104 cells/ml.The cells were randomly divided into 5 groups (n =18 each) using a random number table:control group (group C),in tralipid group (group Ⅰ) and propofol 1,10,100 μmol/L groups (P1-3 groups).In group Ⅰ,10％ intralipid was added to the culture media until the final concentration reached 100 μmol/L.In groups P1-3,propofol was added to the culture media until the final concentration reached 1,10 and 100 μmol/L,respectively,and the cells were then incubated for 3 h.The cell apoptosis was assessed by flow cytometry.The expression of Bcl-2 mRNA and caspase-3 mRNA was detected by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR).The expression of Bcl-2 and actived-caspase-3 protein was determined by Western blot analysis.The apoptosis rate was calculated.Results Compared with group C,the apoptosis rate was significantly increased,the expression of Bcl-2 mRNA and protein was down-regulated,and the expression of caspase-3 mRNA and actived-caspase-3 protein was up-regulated in P1-3 groups (P ＜ 0.05).There was no significant difference in the parameters mentioned above between group Ⅰ and group C (P ＞ 0.05).Conclusion Propofol induces apoptosis in isolated hippocampal neurons by inhibiting Bcl-2 expression and enhancing caspase-3 activity in fetal rats.

Objective To evaluate the effects of propofol on the intracellular calcium ion concentration ([Ca2+ ]i) and nuclear factor kappa B (NF-κB) activity in hippocampal neurons of fetal rats in vitro .Methods Ten pregnant Sprague-Dawley rats at 16-18 days of gestation ,were sacrificed and the fetal rats were taken out from the abdominal cavity .The hippocampal neurons of the fetal rats were isolated and seeded in culture plates .After being cultured for 9 days ,the neurons were divided into 7 groups ( n=12 each ) using a random number table :control group (C group) ,intralipid group (I group) and propofol 0.1 ,1 ,10 ,100 ,1 000 μmol/L groups (P1-5 groups) .In group I ,10% intralipid was added to the culture media until the final concentration reached 100μmol/L .In P1-5 groups ,propofol was added to the culture media until the final concentration reached 0.1 ,1 ,10 , 100 and 1 000μmol/L ,respectively .The cells were then incubated for 3 h .The [Ca2+ ]i and cellular morphology of hippocampal neurons were examined by laser scanning confocal microscopy before incubation with propofol and within 10 min after the end of incubation with propofol .The expression of NF-κB protein in the nucleus was detected at 7 days after the end of incubation with propofol by Western blot analysis to reflect NF-κB activity . Results Propofol increased [Ca2+ ]i in P2-4 groups ,while decreased [Ca2+ ]i in group P5 ( P<0.05 ) .Compared with group C ,the activity of NF-κB was significantly decreased in P1-5 groups ( P<0.05 ) ,while no significant change was found in C and I groups ( P>0.05 ) .The structure of hippocampal neurons was normal in C ,I and P1 groups .The branchings of axons and dendrites in hippocampal neurons were significantly decreased in P 2-4 groups , while the structure of hippocampal neurons became fuzzy , the cell membrane was destroyed and the axons and dendrites were not seen in group P5 .Conclusion Propofol can produce neurotoxic effects on hippocampal neurons of fetal rats by changing the [Ca2+ ]i and promoting NF-κB activation in vitro .

Objective To find out a more rational pathological classification criteria for renal injury in patients with type 2 diabetes mellitus. Methods The renal clinicopathological features of forty-nine type 2 diabetic patients with maeroalbuminuria were collected and were compared retrospectively. The patients without diabetic renal disease were excluded. According to the pathological features, the patients were divided into two groups: typical diabetic glomerulopathy (DG) and atypical diabetes-related renal disease (ADRD). Results The renal biopsy revealed DG accounted for 59.2% of the patients, while the remaining 40.8% presented atypical renal injury defined as ADRD. In DG group, volume fraction of mesangium per glomerulus, glomerular basement membrane width, atrophic tubules index, intersititium injury index and prevalence of hyalinization of renal arteriole were higher; podocyte density per glomerulus was lower; duration of type 2 diabetes was longer; the level of fast blood glucose, systolic blood pressure, mean arterial pressure, proteinuria and prevalence of diabetic retinopathy (DR) were higher; glomerular filtration rate (GFR) was lower. In ADRD group, body mass index and prevalence of obesity were higher; dyslipidemia was more severe. GFR was negatively correlated with glomerular global sclerosis rate in both DG and ADRD group. Proteinuria was positively correlated with volume fraction of mesangium per glomerulus in DG. No correlation between proteinuria and pathological features was found in ADRD. DR (94.8%) and duration of type 2 diabetes over five years (90.7%) had high negative predictive value for DG. Conclusions Renal injuries in type 2 diabetes patients are heterogeneous. ADRD is an atypical renal injury in type 2 diabetes patients whieh is different from DG. DR and duration of diabetes are more helpful in predicting DG separating from ADRD.

Objective To observe the impact of IL-1β on the expression of lectin-like oxidized LDL receptor 1 (LOX-1) and ATP-binding cassette transporter A1 (ABCA1) in human mesangial cell line (HMCL), and its association with cholesterol homeostasis of HMCL. Methods Levels of LOX-1 and ABCA1 of HMCL induced by IL-1β were examined by using real-time PCR and Western blot. Results IL-1β up-regulated LOX-1 mRNA and protein expression. Treated with 5 μg/L IL-1β, the levels of LOX-1 mRNA and protein reached the peak after 6 h and 24 h of stimulation and were 6.87 folds and 1.88 folds of control rspectively. The expression of ABCA1 mRNA and protein of lipid-loaded HMCL was down-regulated by IL-1β Stimulated with 5 μg/L IL-1β the expression of ABCA1 mRNA and protein decreased to the lowest level, 19.0% and 50.62% of the baseline respectively. Conclusions The expression of LOX-1 can be up-regulated while the expression of ABCA1 can be decreased by the stimulation of IL-1β. IL-1β can enhance dyslipidemia and influence the balance of cholesterol homeostasis of HMCL.

Objective To analyze the clinicopathological characteris tics of systemic lupus erythematosus (SLE) with secondary antiphospholid syndrome (APS) . Methods Data of 11 cases of SLE with secondary APS (SLE with APS) admitted to Peking Union Medical College Hospital from January 2000 to March 2010 were retrospectively analyzed. Kidney biopsy was performed on all of these patients. Differences of clinicopathology and outcomes between SLE with and without APS were compared. Results Renal involvement was found in all the SLE with APS patients. The prominent clinical manifestations included hypertension (54.5%), nephrotic level of proteinuria (24 h proteinuria ≥3.5 g)(72.7%) and renal insufficiency (45.5%). Diastolic blood pressure, mean arterial pressure and glomerular filtration rate in SLE with APS were significantly higher than those in SLE without APS (all P＜0.05). In 8 out of 11 cases (72.7%), APS nephropathy (APSN) in kidney biopsy was found, characterized by small vessel vaso-occlusive lesions. These included thrombotic microangiopathy (TMA), fibrous intimal hyperplasia (FIH), focal cortical atrophy (FCA) and tubular thyroidization. Among those, 5 cases (45.5%) had chronic APSN and 4 (36.4%) had acute APSN (one case had acute and chronic APSN at the same time). The incidences of APSN and acute APSN in the SLE with APS group were significantly higher than those in SLE without APS group (P＜0.05). Conclusions The major renal manifestations of SLE with APS are hypertension, nephrotic level of proteinuria and renal insufficiency. Other than lupus nephritis, also a high incidence of APSN is found in SLE with APS patients.

Objective To study the pathologic pattern and clinical feature of type 2 diabetes mellitus complicated with chronic kidney diseases (CKD). Methods Clinicopathological features of 155 type 2 diabetic patients complicated with CKD were collected and analyzed retrospectively. The patients were divided into four groups: typical diabetic glomerulopathy (DG),atypical diabetes-related renal disease (ADRD), non-diabetic renal diseases (NDRD) and DG complicated with NDRD. Results Renal biopsies revealed DG accounted for 18.7% of the patients, ADRD accounted for 12.9%, NDRD accounted for 60.0%, and DG complicated with NDRD accounted for 8.4%. In DG group, duration of type 2 diabetes was longer;the level of fast blood glucose, systolic blood pressure, mean arterial pressure and prevalence of diabetic retinopathy (DR) were higher;proteinuria was heavier and evaluated glomerular filtration rate (eGFR) was lower. In ADRD group, body mass index and prevalence of obesity were higher;dyslipidemia was more severe. Gross hematuria and acute renal insufficiency could be only found in NDRD group.Without DR, duration of diabetes under 5 years, gross hematuria, acute renal insufficiency,evidences of autoimmune diseases and proteinuria≥3.5 g/24 h but eGFR ≥60 ml/min were specific valuable predictors for NDRD. Conclusions Renal injuries in type 2 diabetic patients are structural heterogeneous, in which NDRD is more common and is different from ADRD and DG.Renal biopsy should be considered when type 2 diabetic patients complicated with CKD present at least one characteristic as follows: duration of diabetes under 5 years, without DR, history of gross hematuria, acute decrease of renal function, evidences of autoimmune diseases and proteinuria ≥ 3.5 g/24 h but eGFR ≥ 60 ml/min.

Objective To analyse renal lesions associated with infectious endocarditis (IE). Methods Renal lesions associated with IE were reviewed. One hundred and fifty-five cases of IE were admitted to Peking Union Medical College Hospital from 1983 to 2004. C-square, t-test and Spearman's rank correlation analysis were performed. Results One hundred and thirty-seven(84.4%) cases of renal lesions associated with IE with an average age of 38 were found. The ratio of male to female was 1.4 and the period of pre-renal lesion was 4.8 months. Renal lesions included asymptomatic hematuria and/or proteinuria (71.0%), acute nephritic syndrome (6.5%), nephrotic syndrome (2.6% ), rapid progressive glomerulonephritis (1.3% ), renal embolism (1.3% ), isolated pyuria(3.2%), renal lesion not directly related to IE(2.6%). Acute renal insufficiency in 14 cases were caused by glomerulonephritis (5 cases), acute interstitial nephritis (5 cases), renal embolism(1 case), acute heart failure(5 cases) and the adverse effect of antibiotics (2 cases). Renal biopsy was taken in four patients. One diffuse proliferative glomerulonephritis, one membranous glomerulonephritis, one membrane-proliferative glomerulonephritis and one crescentic glomerulonephritis were found. All patients received antibiotic therapy and three of them stopped taking antibiotics, which was suspected to cause renal lesion. 20.4% cases received surgical therapy. 3.6% were treated with corticosteroid and/or immunoimpressive drugs and two cases of them were treated with intravenous bolus methylprednisolone. One case recieved anticoagulant therapy. 4.5% cases died. 43.8% cases with renal lesions were cured and 85.7% serum creatinine level decreased to normal. Statistical analysis showed that active treatment made no improvement on neither patients with or without renal lesion nor patients with different severity of renal lesion. Conclusions Renal lesions associated with IE are common. Most are asymptomatic hematuria and/or proteinuria. Acute nephritic syndrome, nephrotic syndrome, rapid progressive glomerulonephritis, renal embolism may also occur. It maybe appropriate to treat with corticosteroid, immunopressive drugs or intravenous bolus methylprednisolone for patients with rapid progressive glomerulonephritis under successful management of infective endocarditis.