OBJECTIVE: To investigate the protective effects of the ethanol extract of Radix et Rhizoma Gentianae on acute lung injury (ALI). METHODS: 48 NIH male mice were randomized into blank group (normal saline), model group (normal saline), dexamethasone acetate group (0.005 g?kg-1), ethanol extract of Radix et Rhizoma Gentianae groups (high-dosage of 10 g?kg-1, medium dosage of 5 g?kg-1 and low dosage of 2.5 g?kg-1). The groups were administered via i.g gtt once a day for five days. ALI model was induced by intratracheal injection of lipopolysaccharide (LPS, 2.5 mg?kg-1) after last treatment. ELISA method was applied to determine the content of prostaglandin E2 (PGE2), interleukin-10 (IL-10) and cycloxygenase-2(COX-2) at 24 h after modeling. RESULTS: As compared with model group, the content of PGE2 and COX-2 in ethanol extract of Radix et Rhizoma Gentianae groups were decreased significantly (P

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.

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 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

Paeonia lactiflora Pall. has a long history of utilization, and is widely used in pharmaceutical, food and cosmetic industry. In this paper, the invention patents of traditional medicine P. lactiflora before 2014 were retrieved, and totally 18 192 patent families were obtained. And we formed the patent analysis report of medicine P. lactiflora based on a multi-angle analysis. Results show that, as to the patent number of medicine P. lactiflora, China is much more than any other country, and the applications mainly came from pharmaceutical enterprises. But the technologi-cal quality of patents and international protection ratio are low in our country. We need to strengthen in treatment of cardiovascular and liver disease. The patents mainly focused on the use of tonic health, which is compatible with Traditional Chinese Medicine. On the product development, dosage forms need to be enhanced. As to the compre-hensive utilization of resources, the flower and seed of P. lactiflora have relatively larger research space and value. This work will be helpful for researchers in deeply understanding the research achievements of medicine P. lactiflora, and provides the reference data for the future research.

Objective : To evaluate the quality of neonatal inherited metabolic diseases screening in Chaoyang District, Beijing Municipality from 2012 to 2021, so as to provide insights into improvements in the screening quality and efficiency of neonatal inherited metabolic diseases. Methods: Data pertaining to screening of neonatal inherited metabolic disease in Chaoyang District from 2012 to 2021 were captured from Beijing Center for Neonatal Disease Screening. The percentage of screening, eligible rate of blood smears collection, re-examination rate of suspected cases, and definitive diagnosis of congenital hypothyroidism (CH), phenylketonuria (PKU) and congenital adrenal hyperplasia (CAH) were analyzed to evaluate the quality of neonatal inherited metabolic diseases screening in Chaoyang District. Results: There were 484 002 live neonates in Chaoyang District from 2012 to 2021, and 481 395 neonates were screened for inherited metabolic diseases, with a screening rate of 99.46% and 99.71% eligible rate of blood smears collection. A total of 4 305 suspected positive cases were screened, including 4 148 cases recalled for re-examinations, with a 96.35% re-examination rate of suspected cases, and the re-examination rates of CH, PKU and CAH were 96.37%, 96.79% and 95.65%, respectively. Totally 482 neonates were definitively diagnosed with inherited metabolic diseases, with an overall incidence rate of 1/999, and the incidence rates of CH (307 cases), hyperthyrotropinemia (103 cases), PKU (66 cases) and CAH (6 cases) were 1/1 568, 1/4 674, 1/7 294 and 1/20 233, respectively. Conclusions The screening rate and re-examination rate of neonatal inherited metabolic diseases was both more than 95% in Chaoyang District from 2012 to 2021. Improving the management of neonatal inherited metabolic diseases screening and the recall of suspected cases is required.

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.

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 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.