Objective To investigate the characteristic of 13C-methacetin breath test (13C-MBT) as a tool to monitor the hepatic function of patients with hepatic carcinoma by comparing with Child-Pugh classification and general liver function. Methods Thirty-nine patients with primary liver cancer, 16 patients with hepatic metastasis and 14 healthy volunteers serving as controls were included in this study. According to Child-Pugh classification, the primary liver cancer patients were divided into A, B and C subgroups. All subjects received 13C-MBT and routine liver function tests after an overnight fast. The three major parameters of 13C-MBT i.e. maximum excretion rate before 40 min (Mwnax40), 13CO2 cumulative excretion of 40 min(CUM40) and that of 120 min(CUM120) were recorded and the two metabolism curves (DOB curve, MV curve) were made. Results (1) In the control, hepatic metastasis and primary liver cancer groups, both the DOB curve and MV curve were similar in shape; the peak time occurred at about 20 min after administration and then the curves lowered progressively. There were significant differences between the primary liver cancer group and the other two groups, but it was not statistically different between the hepatic metastasis group and the controls. The shape was obviously distinct in the groups A, B and C of primary liver cancer. The group A had a single sharp peak curve, group B a relatively flat peak curve with a lower level for a long time after the ascending phase and group C no clear excretion peak or even a negative curve. (2) As to the three parameters of 13C-MBT, there were statistical difference between the primary liver cancer group and the other two groups(P <0. 05). Between hepatic metastasis group and controls,there was statistical difference about CUM120 (P <0. 05), but no statistical difference about Mvmax40 and CUM40. While in the three groups with primary liver cancer groups, there was statistical difference between group A and B in Mvmax40 and CUM40 ( P < 0. 05 ), but no statistical difference between group B and C. As to CUM120, there was statistical difference only between group A and C. (3) Comparing the three parameters of 13C-MBT with routine liver function tests, there was negative correlation with TBA, positive correlation with Alb, PA, ChE and no correlation with ALT, AST, TBil, γ-GT, ALP and PT. (4) There was a good consistency between 13C-MBT and Child-Pugh classification in the evaluation of liver function of patients with liver cancer ( Kappa = 0. 647, P < 0. 05 ). Conclusion The value of the three parameters of 13C-MBT is decreased with severity of the liver disease and 13C-MBT may be used to evaluate the reserved hepatic function in patients with primary liver cancer with a diagnostic value equivalent to Child-Pugh classification. The study further confirms that 13C-MBT has correlation with TBA,AIb, PA and ChE.

ObjectiveWith a GcneChip(R) cxpression analysis,98 known gencs and 31 exprcssed sequence tags (ESTs) were found to be differentially expressed between KK/Ta and BALB/c kidneys.To further screen the susceptibility genes for diabetic nephropathy,the temporal and spatial expression patterns of differentially expressed gene-adipsin were investigated.MethodsThe body weight,blood glucose,urinary albumin/creatinine ratio,and renal pathological changes of KK/Ta and BALB/c mice were measured at the 7,20,28 and 36 weeks of age.Total RNA was extracted from the kidney,heart,liver,lung,and brain.The temporal and spatial expression patterns of adipsin in diabetic KK/Ta mice were examined by competitive RT-PCR.The correlation analysis between adipsin expression and albuminuria level was carried out.ResultsThe mRNA expression of adipsin was found in the kidney,heart,lung,and brain,but not in liver.The expression of adipsin in diabetic KK/Ta mice at 20 weeks of age was significantly down-regulated in kidney,heart,and lung than that in age-matched BALB/c mice,and unaltered in brain.Adipsin expression in KK/Ta kidneys was significantly down-regulated with aging and negatively correlated to urinary albumin/creatinine ratio( r =-0.807,P ＜ 0.05).ConclusionsThe expression of adipsin mRNA was downregulated in kidney,heart,and lung in diabetic state.Adipsin expression in KK/Ta kidneys was negatively correlated to urinary albumin/creatinine ratio.It might be a candidate gene for diabetic nephropathy.

Objective To investigate the effects of angiotensin receptor blocker (ARB)losartan on the glomerular protein expression profile of spontaneous type 2 diabetic KKAy mice by two-dimensional differential gel eleetrophoresis and MALDI-TOF mass spectrometry.Methods 8-week-old spontaneous type 2 diabetic KKAy mice were randomly divided into losartan (10 mg·kg-1·d-1 given in drinking water) treatment group and non-treatment group.Eight-week-old C57BL/6 mice were used as normal control.The glomeruli were separated by magnetic bead perfusion through thoracic aorta at age of 20 weeks,then glomerular protein was extracted.The glomerular protein expression profile was investigated by CyDyes minimal fluorescence labelling,two-dimensional differential gel electrophoresis and MALDI-TOF mass spectrometry.Results KKAy mice developed higher body weight and blood glucose,higher urinary microalbumin creatinine ratio at age of 20 weeks than C57BL/6 mice at the same age (all P＜0.05).Losartan treatment markedly reduced urinary microalbumin creatinine ratio [(539.71 ±100.23)mg/g vs (728±177.19) mg/g],attenuated mesangial expansion and the thickening of glomerular basement membrane,but had no effect on the blood glucose.By DeCyder 2-D differential analysis software,62 protein spots of differential expression were found in glomeruli between losartan treatment and non-treatment KKAy mice at age of 20 weeks.Among them,41 proteins were identified by peptide mass fingerprinting.The expressions of 28 proteins were up-regulated by losartan treatment,including glycerokinase,sulfite oxidase,glycine amidinotransferase,adenosylhomocysteinase,etc.The expressions of 13proteins were down-regulaled by losartan treatment,including 3-mercaptopyruvate sulfurtransferase,ATP synthase subunit d,60 000 heat shock protein,stress-70 protein (alternative name 75 000glucose-regulated protein,GRP75),etc.Six differcntially expressed proteins were found in glomeruli between non-treatment KKAy mice and C57BL/6 mice,and the differential expressions were suppressed by losartan treatment,including dihydrolipoyllysine-residue acetyltransferase component of pyruvate dehydrogenase complex,succinyl-CoA ligase (GDP-forming) subunit beta,mitochondrial,ATP synthase subunit d,GRP75,nucleoside diphosphate-linked moiety X motif 19 and seleniumbinding protein 1.Conclusions Losartan significantly reduces the urinary protein excretion rate and renal pathological lesion of spontaneous type 2 diabetic KKAy mice,and suppresses the differential expression of mitochondrial ATP synthase subunit d,GRP75,selenium-binding protein 1,etc in glomeruli.Losartan may play a renoproteetive role by reducing glomerular mitochondrial reactive oxygen species genesis and inhibiting oxidative stress.

Objective To investigate the relationship between blood pressure(BP) and prognosis in three different ischemic stroke subtypes.Methods The consecutive patients with a brain infarction proven on diffusion-weighted MRI who were hospitalized within 48 hours after stroke onset between April 2007 and April 2008 were registered.All subjects with acute ischemic stroke consecutively admitted to the neurological wards of the First Affiliated Hospital of Wenzhou Medical College,were registered in the Wenzhou Stroke Registry Program.Data were collected and coded at primary registration.The BP levels were studied during the initial 7 hospital days.Survival and dependency were assessed at 3 months.Outcomes were adjusted age,consciousness level,admission NIHSS score,the decline level of systolic BP,the decline level of diastolic BP,complication and so on. Logistic regression model was used to estimate the relationship between BP and prognosis.Results A U-shaped effect was observed in each subgroup between BP and prognosis.In the subgroups of atherothrombosis,cardioembolism and small artery disease,those who had a BP of 150/95 mm Hg(1 mm Hg=0.133 kPa)on admission,140/90 mm Hg on day 1-7 would have a better prognosis.In the subgroups of atherothrombosis and cardioembolism,the decrease of BP during the first 24 hours was the independent predictor of the death and disability at 3-month.In the atherothrombosis group,when the decrease of systolic BP during the first 24 hours was greater than 20 mm Hg,the risk of the death and disability at 3-month increased 4.44 times(OR 4.44,95%CI 1.70-11.59,P=0.002).In the atherothrombosis group,when the decrease of diastolic BP during the first 24 hours was greater than 10 mm Hg,the risk of the death/disability at 3-month increased 3.70 times(OR 3.70,95%CI 1.54-8.90.P=0.00).In the cardioembolism group,the risk increased respectively 7.98 times(OR 7.98,95%CI 1.34-47.66.P=0.026)and 6.68 times(OR 6.68.95%CI 1.55-28.79,P=0.01).In the subgroups of small artery disease,the decrease of BP during the first 24 hours was not the independent predictor of the death and disability at 3-month.Conclusions A U-shaped effect is observed in each subgroup between BP and prognosis.In the subgroups of atherothrombosis and cardioembolism,the decrease of BP during the first 24 hours is the independent predictor of the death and disability at 3-month.

Objective To investigate the changes of hepatic function before and after transcatheter arterial chemoembolization (TACE) in primary liver cancer patients with different quantifications and classifications using 13C-methacetin breath test (13C-MBT), and to provide risk predicts for TACE. Methods 28 cases of primary liver cancer patients and 10 cases of metastatic liver cancer patients were selected.General examination items of liver function and 13C-MBT detection were performed on all cases. Primary liver cancer was divided into 4 groups according to the results of 13C-MBT (group 1 was normal or pathological liver damage; group 2 was grade A; Group 3 was grade B; group 4 was grade C). 13C-MBT detection was carried out in week 1 and 4 after TACE. Results The three parameters of 13C-MBT is 13CO2 maximum excretion rate before 40 min/27 (MVmax40), 13CO2 cumulative excretion of 40 min/12 (CUM40), and than of 120 min/28(CUM120). Compared with the parameters [(0.628±0.191)%, (0.628±0.289)%, (0.577±0.286)%] before TACE in hepatic metastasis, there were no statistic effect between week 1 [(0.600±0.187)%, (0.559±0.189)%, 0.587±0.181)%] and week 4 [(0.700±0.230)%, (0.734±0.229)%, (0.724±0.252)%] after TACE. Conclusion The lower classification of 13C-MBT, the more impairment to liver function after TACE. This could provide an important diagnostic basis for TACE.

Objective A total of 102 ESRD patients undergoing hemodialysis for over 3 months in our dialysis center were asked to complete the SF-36 scales.They were also assessed by Hamilton Depression Scale(HAMD17).Univariate analysis was performed to determine the impact of factors such as age,gender,employment status,education,mental status and dialysis status on their quality of life.Methods The 102 patients with ESRD undergoing hemodialysis for over 3 months in our dialysis center completed the SF-36 scales with self-administration,and they were also assessed by Hamilton Depression Scale (HAMD17).Univariate analysis was performed to determine the impact of variables such as age,gender,employment status,education,mental status and dialysis status on the quality of life in patients.Results The scores of PF,RP,BP,GH,VT,SF,RE and MH in patients were significantly lower than those in the normal controls(P

Objective To explore the mechanism of up-regulation of tubular liver-type fatty acid binding-protein (L-FABP) in IgA nephropathy (IgAN) and its renoprotective role.Methods Murine mesangial cells (MCs) from primary cell culture were cultured with aggregated IgA (AIgA) (10 to 250 mg/L) for 48 hours. The supernatant after culture was collected as AIgA-MC medium. Murine proximal tubular cell line (mProx) stably expressing human L-FABP (hL-FABP) by transfection (mProx-L) were cultured with AIgA, AIgA-MC medium and /or neutralizing anti-TNF-α antibody and recombinant murine TNF-α, respectively. AIgA-MC medium (AIgA final concentration was 25 mg/L) was cultured with mProx and mProx-L cells. The mRNA expressions of hL-FABP and MCP-1 of the cells were detected by real-time PCR. The protein expressions of hL-FABP and 4-HNE of the cells were detected by Western blotting. Results (1) The hL-FABP mRNA and protein expression stimulated by AIgA-MC medium was significantly higher as compared to AIgA (P＜0.01). (2) Pre-incubation of neutralizing anti-TNF-α antibody (final concentration was 1 and 5 mg/L) with mProx-L cells could significantly suppress the up-regulation of hL-FABP protein expression induced by AlgA-MC medium (P＜0.05 and P＜0.01).(3) Recombinant murine TNF-α (final concentration was 50 and 250 ng/L) also induced a significant up-regulation of hL-FABP expression (P＜0.01). (4) After the stimulation of AIgA-MC medium, both 4-HNE protein expression and MCP-1 mRNA expression were significantly suppressed in mProx-L cells compared to those of mProx cells (P ＜0.05 and P＜0.01). Conclusion Mesangial cell-derived TNF-α can induce up-regulation of tubular L-FABP expression. Overexpression of tubular L-FABP may lessen the progression of IgAN by reducing oxidative stress and inflammatory mediators.

Objective To identify the candidate genes in the vicinity of a susceptibility locus (urinary albumin 1,UA-1) contributing to the development of albuminuria in type 2 diabetic KK/Ta mice. Methods Total RNA was extracted from the kidneys of KK/Ta (n=3) and BALB/c (n=2) mice at 20 weeks of age.The gene expression profile in kidney was investigated using the Affymetrix Murine Genome U74Av2 array.Competitive RT-PCR was used to confirm the differential expression of syndecan-4 which located in the vicinity of UA-1.Genome DNA was extracted from KK/Ta and BALB/c mice.DNA sequence analysis of the coding and promotor region of syndecan-4 gene was conducted. Results In the vicinity of the susceptibility locus (UA-1)contributing to the development of albuminuria in type 2 diabetic KK/Ta mice,10 candidate genes that showed differential expression were identified.Among them,the gene expression of syndecan-4in KK/Ta kidneys at 20 weeks of age was up-regulated by 26.1 times of age-matched BALB/c kidneys.Sequence analysis revealed two synonymous polymorphisms in the coding region (A93C and T216C) and three polymorphisms in the promoter region (-T263C,-T396C and -G669A) of the syndecan-4 gene.The TATA box was found at 321 bp upstream from the transcription start site,and the T263C polymorphism was located in the binding site of transcription factor Clox.Conclusions Syndecan-4 gene is mapped in the vicinity of the susceptibility locus contributing to the development of albuminuria in type 2 diabetes.The gene expression of syndecan-4 in KK/Ta kidneys is up-regulated than that in age-matched BALB/c kidneys at 20 weeks of age.Thus syndecan-4 may be one of the potential candidate genes responsible for diabetic nephropathy.Sequence differences in the promoter region influence the expression levels of syndecan-4 genes in KK/Ta kidneys.

Objective To identify susceptible miRNAs for the pathogenesis of diabetic nephropathy (DN) and the molecular targets of losartan treatment. Methods The 8-week age KKAy mice were divided into losartan treatment group (10 mg· kg-1· d-1) and non-treatment group,C57BL/6 mice were used as the control group.At age of 20 weeks,body weight,random blood glucose,urinary albumin and urinary creatinine were tested,and kidney morphology was observed.Glomeroli were separated by magnetic beads perfusion,and total RNA were extracted.MiRNAs expression profiles were analyzed by the Affymetrix GeneChip miRNAs arrays. Results At age of 20 weeks,KKAy mice developed higher body weight,higher blood glucose and higher urinary microalbumin creatinine ratio than C57BL/6 mice,and the glomerular basement membrane thickened,mesangial matrix widened.Losartan treatment markedly improved the level of urinary albumin creatinine ratio [(539.71±100.23) mg/g vs (728±177.19) mg/g,P＜0.05)] and pathological lesion of KKAy mice.The miRNA array analysis showed that there were 22 miRNAs differentially expressed between KKAy non-treatment mice and C57BL/6 mice glomeruli at age of 20 weeks.Among them,10 miRNAs were up-regulated,and 12 miRNAs were down-regulated.The expression of 4 miRNAs was down-regulated in glumeruli of KKAy mice treated by losartan compared with that of non-treatment mice.The expressions of miRNA-503 and miRNA-181d were significantly up-regulated in the glumeruli of KKAy mice and inhibited by losartan treatment, Conclusion The expressions of miRNA-503 and miRNA-181d are significantly up-regulated in the glumeruli of KKAy mice and inhibited by losartan treatment,which may be new therapeutic targets of DN.

Objective To investigate the renoprotection of tubular L-FABP in murine IgA nephropathy (IgAN) induced by bone marrow transplantation(BMT). Methods IgAN models were reconstituted by BMT from IgAN-prone mice into mice (Tg) transgenically tubular overexpressing human L-FABP (hL-FABP) and wild type (WT) mice. These recipients were sacrificed at 6 and 12 weeks after BMT and their kidneys were collected. The expressions of hL-FABP, fibronectin (FN)and monocyte chemoattractant protein-1 (MCP-1) mRNA were detected by real-time PCR. hL-FABP,FN, type Ⅳ collagen (Col Ⅳ ), hemeoxygenase-1 (HO-1) and 4-hydroxy-2-nonenal (4-HNE)modified proteins were detected by Western blotting. The distribution of hL-FABP and FN protein in kidney was detected by immunohistochemistry. The level of serum IgA, urinary albumin and urinary hL-FABP was detected by ELISA. Results (1) IgAN was reconstituted in both Tg and WT mice by BMT: mesangial IgA deposition and up-regulation of serum IgA. The levels were not significantly different between two groups (Tg-ddY and WT-ddY). (2) hL-FABP was expressed in proximal tubular cells of normal Tg mice. The mRNA (1.62±0.32 vs 0.46±0.09, P＜0.01) and protein expression (1.74±0.76 vs 1.14±0.31, P＜0.01) of hL-FABP was up-regulated in Tg-ddY kidney and urinary hL-FABP level (μg/g creatinine) was significantly increased (59.87±26.75 vs 31.01±14.86, P＜0.05) at the 6th week after BMT. (3) WT-ddY mice showed a significantly higher urinary albumin level (mg/L) (828±656 vs 82±22, P＜0.01), severer mesangial matrix expansion (P＜0.01),more glomerular FN and Col Ⅳ deposition at the 12th week. (4) Up-regulation of renal hL-FABP was associated with significant suppression of renal HO-1 expression (P ＜0.05),accumulation of 4-HNE modified proteins (P＜0.05) and MCP-1 mRNA expression (P＜0.01) in Tg-ddY mice. Conclusion Tubular L-FABP may lessen the progression of glomerular damage at early stages of IgAN by reducing oxidative stress and inflammatory mediators.