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Objective:To observe the expression of angiopoietin-like protein 4 (ANGPTL4) signaling pathway in adenine-induced chronic kidney disease (CKD) rat model, and to explore the role of this pathway in renal fibrosis.Methods:Thirty-six male Sprague-Dawley rats were randomly divided into control group (saline, intragastric administration, n=15) and CKD group (250 mg·kg -1·d -1 2.5% adenine, intragastric administration, n=21). At the end of the 1st, 2nd and 4th week, 5 rats were randomly selected from each group. Renal function and 24-hour urinary protein quantity were measured. HE and Masson staining were used to observe the pathological changes of kidneys. Immunohistochemistry and real-time PCR were used to detect renal protein and mRNA expressions of hypoxia-inducible factor-1α (HIF-1α), ANGPTL4, bone morphogenetic protein 7 (BMP7), Smad1, α-smooth muscle actin (α-SMA) and type Ⅰ collagen (Col-Ⅰ). Pearson correlation analysis was used to analyze the correlation between the different indicators. Results:(1) Compared with the control group, the expression levels of serum creatinine and blood urea nitrogen in CKD group were higher at each time point, and the expression levels of 24-hour urinary protein quantity were higher at the end of the 2nd and 4th week (all P < 0.05). (2) HE and Masson staining showed that there were obvious renal structural disorders and collagen fiber deposition at each time point in CKD group compared with the control group, which got worse with time. (3) The results of immunohistochemistry and real-time PCR showed that compared with the control group, the protein and mRNA expression levels of ANGPTL4, α-SMA and Col-Ⅰ were higher, while the protein and mRNA expression levels of BMP7 and Smad1 were lower at the end of the 1st, 2nd and 4th week, and the protein and mRNA expression levels of HIF-1α were higher at the end of 2nd and 4th week in CKD group (all P < 0.05). (4) Correlation analysis results showed that HIF-1α and ANGPTL4 mRNA expression were positively correlated with α-SMA mRNA ( r=0.919, P < 0.001; r=0.757, P < 0.001), and also positively correlated with Col-Ⅰ mRNA ( r=0.925, P < 0.001; r=0.777, P < 0.001). HIF-1α mRNA expression was positively correlated with ANGPTL4 mRNA ( r=0.766, P < 0.001). There were significant negative correlations between HIF-1α, ANGPTL4 mRNA and BMP7 mRNA ( r=-0.652, P < 0.001; r=-0.741, P < 0.001). Conclusions:ANGPTL4 signaling pathway may be activated in adenine-induced CKD rat model, and involved in the renal fibrosis process of CKD.
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OBJECTIVE@#To establish a prediction model of acute kidney injury (AKI) in moderate and severe burn patients, so as to provide basic research evidence for early identification of burn-related AKI.@*METHODS@#Patients who were admitted to the department of plastic burn surgery of the Affiliated Hospital of Southwest Medical University from November 2018 to January 2021 were selected, and their clinical characteristics, laboratory examinations and other indicators were recorded. Multivariate Logistic regression analysis was used to screen out the risk factors of AKI related to moderate and severe burns, and R software was used to establish the nomogram of moderate and severe burn patients complicated with AKI. The Bootstrap method model was used for internal verification by repeating sample for 1 000 times. Consistency index and calibration curve were used to evaluate the accuracy of the model, and the receiver operator characteristic curve (ROC curve) and the area under the curve (AUC) were used to evaluate the prediction efficiency, decision curve analysis (DCA) was used to evaluate the clinical utility of the model.@*RESULTS@#A total of 186 patients with moderate and severe burn were included, among which 54 patients suffered from AKI, and the incidence rate was 29.03%. Multivariate Logistic regression analysis showed that the total burn surface area [TBSA; odds ratio (OR) = 1.072, 95% confidence interval (95%CI) was 1.031-1.115, P = 0.001], estimated glomerular filtration rate (eGFR; OR = 0.960, 95%CI was 0.931-0.990, P = 0.010), neutrophil (NEU; OR = 1.190, 95%CI was 1.021-1.386, P = 0.026), neutrophil/lymphocyte ratio (NLR; OR = 0.867, 95%CI was 0.770-0.977, P = 0.019), D-dimer (OR = 4.603, 95%CI was 1.792-11.822, P = 0.002) were the risk factors for patients with moderate and severe burn complicated with AKI. Taking the above indexes as predictive factors, a nomogram prediction model was established, the ROC curve was plotted with AUC of 0.998 (95%CI was 0.988-1.000). Optimum threshold of ROC curve was -0.862, the sensitivity was 98.0% and the specificity was 98.2%, and the consistency index was 0.998 (95%CI was 0.988-1.000). The calibration curve showed that the prognostic nomogram model was accurate, DCA showed that most patients can benefit from this model.@*CONCLUSIONS@#The burned patients with higher TBSA, NEU, NLR, D-dimer and lower eGFR tend to suffer from AKI. The nomogram based on the above five risk factors has high accuracy and clinical value, which can be used as a predictive tool to evaluate the risk of AKI in moderate and severe burn patients.
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Humanos , Prognóstico , Nomogramas , Estudos Retrospectivos , Queimaduras/complicações , Injúria Renal Aguda/etiologia , Curva ROCRESUMO
Objective:To analyze the changes in serum metabolites of patients with uremia using ultra-high performance liquid chromatography-mass spectrometry (UHPLC-MS), and provide a theoretical basis for the prevention and treatment of uremia.Methods:Uremia patients from the Department of Nephrology, the Affiliated Hospital of Southwest Medical University, and the volunteers from the Health Examination Center were enrolled in this study. According to the inclusion and exclusion criteria, 20 uremia patients (experimental group) and 20 volunteers (control group) were screened out. UHPLC-MS was used to detect the metabolites in the serum of subjects from the two groups, and difference analysis was made to screen the different metabolites, followed by correlation analysis and pathway enrichment study.Results:A total of 412 metabolites were identified by UHPLC-MS. Principal components analysis (PCA) proved that these metabolites could distinguish the control group and the experimental group well. The criteria [variable importance for the projection (VIP)>1, fold changes (FC)>1.25 or FC<0.8 and P value<0.05] was set to screen those significantly different metabolites. Finally, 28 significantly different metabolites were screened out, of which 18 metabolites increased significantly, the other 10 different metabolites decreased significantly. Correlation analysis results proved a certain correlation among 28 different metabolites and the experimental group and control group samples, and between the 28 differential metabolites themselves. Enrichment analysis found that 28 different metabolites might enrich the catecholamine biosynthetic pathway, and pathway analysis suggested that 28 different metabolites might affect glutamate, aspartame acid and glutamate metabolic pathways. Conclusion:Based on metabonomic analysis, some metabolites in the serum of patients with uremia have changed, which can affect some metabolic pathways, thus affecting the pathophysiological process of patients with uremia.
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Objective:To observe the expression of hypoxia-inducible factor 1 alpha (HIF-1α) signaling pathway in the aorta of chronic kidney disease (CKD) rats with vascular calcification and to explore the role of this pathway in aortic calcification of CKD.Methods:Forty 8-week-old male SD rats were randomly divided into control group (CON group, n=15) and CKD with aortic calcification group (CKD+AC group, n=25). The rats were sacrificed at the end of 4 th, 6 th and 8 th week respectively and urine, blood, aorta and kidney samples were collected. The level of serum HIF-1α was tested by ELISA. The pathology changes of kidney were observed by HE staining. The aortic calcification was evaluated by alizarin red staining and calcium content detection. Immunohistochemistry and real-time PCR were applied to detect the protein and mRNA expression of alpha-smooth muscle actin (α-SMA), Runt-related transcription factor 2 (Runx2), HIF-1α, vascular endothelial growth factor A (VEGFA) and Notch1 in the aorta. Results:Compared with CON group, serum urea, creatinine, cystatin C, phosphorus, calcium-phosphorus product and 24-h urine protein were significantly higher in CKD+AC group (all P<0.05). Serum HIF-1α levels were higher at 4 th and 8 th week in CKD+AC group than that in CON group (both P<0.05). There was no significant calcium deposit in the aorta of the CON group at all time points, and calcium deposits were seen in the aorta of the CKD+AC rats at each time point, which gradually increased with time. Compared with CON group, the expressions of aortic α-SMA protein and mRNA were significantly decreased in CKD+AC group at each time point, however the protein and mRNA expressions of Runx2, HIF-1α, VEGFA and Notch1 in the aorta of CKD+AC group rats were markedly increased at each time point (all P<0.05). Correlation analysis showed that the aortic calcium content was positively correlated with serum HIF-1α ( r=0.706, P<0.001) and the protein expressions of HIF-1α ( r=0.852, P<0.001), VEGFA ( r=0.747, P<0.001) and Notch1 ( r=0.813, P<0.001) in aorta. Conclusion:The HIF-1α-VEGFA-Notch1 signaling pathway is activated during aortic calcification in CKD rats, suggesting that this signaling pathway might be involved in the vascular calcification in CKD, and serum HIF-1α is expected to be one of serum markers for CKD vascular calcification.
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Objective:To evaluate the value of 68Ga-1, 4, 7, 10-tetraazacyclododecane-1, 4, 7, 10-tetraacetic acid (DOTA)-fibroblast activating protein inhibitor (FAPI)-04 PET/CT on assessing different pathological grades in patients with renal fibrosis. Methods:A total of 25 patients (11 males, 14 females; age (39.3±13.9) years) diagnosed with renal fibrosis in the Affiliated Hospital of Southwest Medical University from September 2020 to August 2021 were retrospectively analyzed. All patients underwent renal puncture examination and 68Ga-DOTA-FAPI-04 PET/CT examination. The pathological results of kidney puncture were as the " gold standard" to divide the patients into mild fibrosis (Ⅰ), moderate fibrosis (Ⅱ), and severe fibrosis (Ⅲ). At the same time, 20 patients (10 males, 10 females; age (47.5±13.2) years) who underwent 68Ga-DOTA-FAPI-04 PET/CT examination showed no abnormal uptake of radioactivity in bilateral kidneys and no history of urinary system related diseases were enrolled as normal controls. Parameters including the maximum standardized uptake value (SUV max) of both kidneys, the mean standardized uptake value (SUV mean) of the liver, target/background ratio (TBR), glomerular filtration rate (GFR), and serum creatinine (Scr) were collected. Kruskal-Wallis rank sum test and Bonferroni correction method were used to compare the differences of SUV max, SUV mean, TBR and Scr among groups. One-way analysis of variance and least significant difference (LSD) t test were used to compare the difference of GFR among groups. The receiver operating characteristic (ROC) curve analysis was used to analyze the diagnostic efficacy of SUV max for the degree of renal fibrosis. Results:Of 25 patients, 22 had increased imaging agent uptake and the sensitivity of 68Ga-DOTA-FAPI-04 PET/CT in diagnosing renal fibrosis was 88%. The SUV max and TBR of patients with fibrosis grade Ⅰ, Ⅱ and Ⅲ were significantly higher than those of controls (SUV max: 4.40(3.30, 4.50), 5.90(4.28, 6.48), 8.50(7.50, 9.73) and 1.44(1.38, 1.68); TBR: 6.340±2.389, 8.097±1.420, 11.343±2.002 and 2.986±0.645; H values: 33.685, 32.368, all adjusted P<0.05 (Bonferroni correction method)). The Scr of patients with fibrosis grade Ⅰ and Ⅲ were significantly different (70.1(55.4, 92.5) and 174.1(161.4, 498.2) μmol/L; H=9.770, adjusted P<0.05 (Bonferroni correction method)). The liver SUV mean of patients with renal fibrosis grades Ⅱ and Ⅲ were significantly higher than that of controls (0.673±0.129, 0.751±0.170 and 0.514±0.142; H=15.609, both adjusted P<0.05 (Bonferroni correction method)). The GFR of patients with fibrosis grade Ⅲ had significant differences with grade Ⅰ and Ⅱ ((27.867±15.747), (87.756±31.657) and (63.160±29.556) ml/min; F=8.298, both P<0.05). ROC curve analysis showed that the area under curve was 0.946 7 (95% CI: 0.899 6-0.993 8, P<0.001). Conclusion:68Ga-DOTA-FAPI-04 PET/CT has a certain value in evaluating the degree of renal fibrosis, which can be used as a supplementary examination method for diagnosing renal fibrosis.
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Objective @#To explore the candidate genes and potential molecular mechanisms of anti -neutrophil cyto- plasmic antibodies ( ANCA) -associated vasculitis by bioinformatics and experimental validation , and to provide a scientific theoretical basis for the treatment of potential inflammatory targets for ANCA-associated vasculitis .@*Methods@#The GSE108109 chip data was retrieved from the Gene Expression Omnibus (GEO) database , and the differential genes were processed , analyzed and screened using the R language related program package . Kyoto encyclo- pedia of genes and genomes (KEGG) and gene ontology (GO) enrichment analysis was carried out using DAVID online network cable , and the interaction network of the protein encoded by the selected genes of inflammatory syn- drome was constructed through STRING web site . Further endogenous competitive RNA ( ceRNA) regulatory net- work was predicted and constructed through miRWalk and DIANA-LncBase databases , and key genes were screened from the network to draw ROC curve . The renal biopsy samples of patients with ANCA-associated vasculi- tis confirmed by our hospital were collected as the experimental group , and the renal biopsy samples of IgA ne- phropathy and micro-adaptive nephropathy were collected as the control group . Immunohistochemical staining was performed on the collected renal biopsy samples , and the average optical density was calculated by semi -quantita- tive analysis of immunohistochemical staining to further verify the expression of the key genes screened by the bioin- formatics analysis . Pearson linear correlation analysis was performed between the average optical density results and the clinical inflammatory data of patients . @*Results @#846 differential genes were screened , of which 444 genes were significantly up-regulated and 402 genes were significantly down-regulated . Through KEGG and GO analysis , im- portant differentially expressed genes related to inflammation regulation were obtained . Among them , CSF1R and TNFRSF1B , two differentially expressed genes never reported in ANCA-associated vasculitis , attracted our atten- tion . At the same time , we constructed multiple ceRNA regulatory axes including KCNQ1OT1 -hsa-miR-125 a-5p- TNFRSF1B . There were 15 samples of ANCA-associated vasculitis , 6 samples of IgA nephropathy , and 3 samples of micropathological kidney . Immunohistochemical results of renal biopsy specimens showed that the expression of CSF1R and TNFRSF1B in ANCA-associated vasculitis kidney tissue was higher than that in the control group . Pearson correlation analysis of clinical data of patients in ANCA group showed that the expression of CSF1R was positively correlated with the content of neutrophil count ( r = 0. 587) , and the expression of TNFRSF1B was posi- tively correlated with the content of serum C -reactive protein ( r = 0. 646) . @*Conclusion @#Key genes related to in- flammatory regulation such as CSF1R and TNFRSF1B were investigated by bioinformatics methods , and a rigorous ceRNA regulatory network was constructed . The expression of CSF1R and TNFRSF1B in ANCA vasculitis was high- er than that in the control group through immunohistochemistry . The results provides a scientific theoretical basis for the molecular mechanism of inflammation , and laid a good foundation for new therapeutic targets of ANCA-related vasculitis for inflammation .
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Objective To observe the differential expression of high mobility group box - 1 protein (HMGB1) in renal tissues of heat stroke mice models, and to explore its role in the pathogenesis of heat stroke associated acute kidney injury(HS-associated AKI). Methods According to random number table, 20 healthy male C57BL/6J mice were randomly divided into 2 groups, including normal control (n=10) and heat stroke group (n=10). The mice in heat stroke group were given with a 2-hour-exposure in biological simulation chamber (temperature 41℃, humidity 70％). Heat stroke was defined as anal temperature lasting more than 40 degrees Celsius. A 18F - deoxyglucose nuclide labeled vivo imaging was conducted with micro - positron emission tomography(PET)/computer tomography (CT). Serum creatinine was examined with blood example. In order to evaluate the pathological changes, HE stain was conducted with kidney tissue, and mitochondrial morphological changes in kidney tissue were observed by transmission electron microscopy. The expressions of HMGB1 and apoptosis inducing factor mitochondria associated 2 (Aifm2) were examined by immunohistochemical method, and the levels of HMGB1 and RAGE were examined by Western blotting. The cell apoptosis of renal tissue was detected by terminal deoxynucleotidyl transferase -mediated dUTP - biotin nick end labeling assay (TUNEL). The metabolomics of kidney tissue in mice were detected by liquid chromatography - mass spectrometry (LC - MS), and the pathway enrichment analysis was carried out by KEEG database. Results (1) The body temperature of the mice in heat shock group was significantly higher than that in normal control group 45 min after model establishment (P＜0.05). The level of serum creatinine in heat shock group was significantly higher than that in normal control group (P＜0.05), and the levels of 18F - deoxyglucose increased in skeletal muscle and visceral tissue of the mice in heat - shock group. (2) HE staining showed hemorrhage in collecting duct and tubular endothelial cell swelling, and mitochondrial swelling and deformation were observed by transmission electron microscopy in kidney tissue of the heat shock group. (3) Immunohistochemical method showed that the levels of Aifm2 and HMGB1 in heat shock group were higher (P＜0.05). (4) Western blotting showed that the levels of HMGB1 and RAGE in heat shock group were higher than those in normal control group (P＜0.05). (5) TUNEL showed that the number of cells with positive stain in kidney tissue of the heat shock group was higher than that in normal control group (P＜0.05). (6) Between normal control group and heat shock group, 136 differential metabolites were detected in kidney tissues. After analysis by KEGG database, pathway abnormalities such as unsaturated fatty acid metabolism disorder may be associated with HS - associated AKI, and many differential metabolites such as adrenic acid may be important regulatory points in the pathogenesis. Conclusion Acute kidney injury is a common complication of heat shock. It may be related to the dysfunction of renal mitochondria and activation of apoptotic pathway caused by systemic hypercatabolism, which may be related to the disorder of unsaturated fatty acid metabolism and activation of HMGB1. Some differential metabolites may be of high value in HS- associated AKI studies.
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Objective To analyze the clinical features and risk factors of death in patients with acute mushroom poisonings. Methods The clinical data of 210 patients with acute mushroom poisoning admitted to the Affiliated Hospital of Southwest Medical University from July 2013 to December 2016 and received follow-up for at least 6 months were retrospectively analyzed. The data included gender, age, hospitalization time, toadstool features, incubation period, clinical performance, laboratory indicators, and prognosis. According to the prognosis, the patients were divided into survival group and non-survival group, the clinical characteristics and organ or system involvement of the two groups were analyzed, and the risk factors of death in patients with acute mushroom poisoning were explored by univariate and Logistic regression analysis. Results All 210 patients were enrolled in the final analysis, with 172 patients (81.9%) in survival group, and 38 (18.1%) in non-survival group. Patients with an incubation period of 6-24 hours had the highest mortality [15.2% (32/210)]. Most toadstools were in white, red or yellow, with an intake of 20-500 g. More than 85% of patients had gastrointestinal reactions, and liver damage was the most common [58.1% (122/210)] in all patients. The patients with heart and nervous system damage had higher mortality [61.4% (27/44) and 61.3% (19/31)], and the more organs or systems involved, the higher the mortality was. Univariate analysis showed that incubation period ≥ 6 hours, white blood cell (WBC) ≥12×109/L, alanine aminotransferase (ALT)≥200 U/L, aspartate aminotransferase (AST) ≥ 200 U/L, lactate dehydrogenase (LDH) ≥ 500 U/L, prothrombin time (PT) ≥ 20 s, activated partial thrombin time (APTT) ≥ 40 s, prothrombin activity (PTA) ≤ 60%, Na+≤ 135 mmol/L, MB isoenzyme of creatine kinase (CK-MB) ≥ 5 μg/L and myoglobin (Mb) ≥ 100 μg/L were the risk factors of death in patients with acute mushroom poisoning. Multiple factors Logistic regression analysis showed that APTT ≥ 40 s had the greatest lethal risk and could increase the risk of death by 5.35 times [odds ratio (OR) = 6.35, 95% confidence interval (95%CI) = 1.24-32.44], indicating that APTT was an independent risk factor of death in patients with acute mushroom poisoning. Conclusions The mortality of acute mushroom poisoning was high, and liver was the mainly involved organ. The incubation period, WBC, ALT, AST,LDH, PT, APTT, PTA, Na+, CK-MB and Mb could be early indicators to evaluate the prognosis in patients with acute mushroom poisoning, and patients with APTT ≥ 40 s had the greatest lethal risk.
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Objective To observe the expression of Notch1 signaling pathway in the aorta of chronic kidney disease (CKD) rats with vascular calcification and to explore the role of this signaling pathway activation in aortic calcification of CKD.Methods A total of 40 male SD rats were randomly divided into normal group (Nor group) and CKD with vascular calcification group (CKD+VC group).Rats in each group were sacrificed at 4 weeks,6 weeks and 8 weeks respectively after the success of modeling.Their 24-hour urine was reserved to test 24 hour urine protein (24 h Upro);blood sample was collected from abdominal aorta to test blood urea nitrogen (BUN),serum creatinine (Scr),Ca and P.The histopathology of renal was detected by HE staining.The aortic calcification was detected by alizarin red S staining.Immunohistochemistry (IHC) was used to test the protein expressions of alpha-smooth muscle actin (or-SMA),Runt-related transcription factor 2 (Runx2),Notchl,recombination signal-binding protein for immunoglobulin kappa J region (RBP-Jκ),Msh homeobox 2 (Msx2),Jagged1 and Notch1 intracellular domain (N1-ICD) in the aorta.Real time PCR was applied to detect the mRNA expressions of α-SMA,smooth muscle 22 alpha (SM22α),Runx2,alkaline phosphatase (ALP),Notch1,RBP-Jκ,Msx2 and Jagged1.Results Compared with those in Nor group,24 h Upro,BUN and Scr increased in the CKD+VC group at 4th,6th and 8th weeks (all P ＜ 0.05).Numerous continuous calcified nodules were detected in the vascular wall of the CKD+VC group,but none in Nor group.As compared with Nor group,the expression of α-SMA was low,while the expression of Runx2 was relatively high in the CKD+VC rats at each time point (all P ＜ 0.05).The expressions of Notchl,RBP-Jκ,Msx2,Jaggedl and NI-ICD in the Nor group were slightly appeared in the aortic wall,while in CKD+VC group these signal protein expressions increased relatively during the experimental period (all P ＜ 0.05).As compared with Nor group,the expressions of α-SMA and SM22α mRNA were low,yet the expressions of Runx2 and ALP mRNA were high in the CKD+VC rats at each time point (all P ＜ 0.05).The mRNA levels of Notch1,RBP-Jκ,Msx2 and Jagged1 in the CKD+VC group at each time point were significantly up-regulated as compared with the Nor group (all P ＜ 0.05).Conclusions There exist phenotypic changes in smooth muscle cells and activations of Notch1/RBP-Jκ/Msx2 signaling pathway in CKD rats with vascular calcification.It may be one of the important signal transduction pathways.
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Objective To explore the effects of renal artery calcification on the renal function in type 2 diabetic ne-phropathy rats .Methods Rats were randomly divided into control group ( CON group ) , diabetic nephropathy group ( DN group) and DN with vascular calcification group ( DN+VC group) .Rats of group DN and DN +VC were fed with high sugar and fat diet and injected with streptozotocin (STZ)into abdominal cavity to induce type 2 diabetes. After diabetic models were made , rats of group DN+VC were treated by vitamin D 3 plus nicotine .The rats were sacrificed at 8 , 12 and16 week respectively and the pathologic change to the renal artery were microscoped by von Kossa staining .The calcium content were detected by calcium assay kit and double immunofluorescence staining and real-time polymerase chain reaction ( RT-qPCR) were applied to detect the protein and gene expression levels of BMP2 in the renal artery.Measure the levels of blood urea nitrogen (BUN),serum creatinine (Scr),cystatin C (Cys C) and 24 hour urinary protein (24-h UA)respectively at the 8th,12th and 16th weeks.Histopathology of kidney was assessed by hematoxylin/eosin staining .Results The deposition of black granules , the calcium content and the protein and gene expression levels of BMP 2 in DN group were significantly higher than those in group CON and lower than DN+VC group at each time points(P<0.05).The BUN, Scr, Cys C and 24-h UA in group DN and group DN+VC were gradually increased in 8th,12th and 16th weeks, and were higher than those in group CON( P<0.05 ) .Compared with the DN group , only the level of Cys C at each time point and the level of 24-h UA in 16th week in DN+VC group were significantly higher ( P<0.05 ) .The pathological damages of the kidney in group DN showed a continual worsening trend and the pathological changes of the kidney in group DN +VC were more serious than group DN .Calcium content was positively correlated with the increased serum BUN , Scr, Cys C, 24-h UA and BMP2 mRNA ( all P<0.01 ) .Conclusions The occurrence and severity of renal artery calcification may participate in and promote the progression of DN .
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Objeetive To explore the effects of renal artery calcification on the progression of diabetic nephropathy (DN),the activation and its role of bone morphogenetic protein 2(BMP2) signal pathway in renal artery of rats.Methods Sixty male SD rats were randomly divided into control group(CON group),DN group and DN with vascular calcification group (DN+VDN group).Rats of group DN and DN + VDN were fed with high sugar and fat diet and injected with streptozocin (STZ) into abdominal cavity to induce diabetes.After diabetic models were successfully made,rats of group DN+ VDN were treated by vitamin D3 plus nicotine.The rats were sacrificed at 8th,12th and 16th week respectively and the levels of renal function,blood glucose and 24 h urinary protein (24-h Upro) were measured.The pathologic changes to the renal artery were observed by yon-Kossa staining and the calcium content was detected by calcium assay kit.The pathologic changes to the kidney were observed by HE.Immunohistochemistry was applied to detect the protein expression of BMP2/Smad1/Runx2/ Osterix signal pathway in the renal artery and real-time PCR were applied to detect the mRNA expression levels of BMP2 and Runx2.Results The calcium content and the deposition of black granules in DN group were significantly higher than those in group CON and lower than DN + VDN group at each time point (P ＜ 0.05).The renal function indices in group DN and group DN+VDN were gradually increased in 8th,12th and 16th weeks,and were higher than those in group CON (P ＜ 0.05).Compared with that in DN group,although the level of BUN,Scr,Cys C and 24-h Upro in DN+VDN group rats were higher at different time point,the level of Cys C at each time point and the level of 24-h Upro in the 16th week showed significant differences (P ＜ 0.05).The pathological damages of the kidney in group DN and DN+VDN showed a continual worsening trend and the pathological changes of the kidney in group DN+VDN were more serious than those in group DN.Furthermore,the levels of BMP2/Smad1/Runx2/Osterix signal protein and BMP2,Runx2 mRNA in DN rats were higher than those in CON group,lower than DN+VDN group at each time point (P ＜ 0.05).Correlation analysis demonstrated that calcium content was positively correlated with serum BUN,Scr,Cys C,24-h Upro and the expression of BMP2,Runx2 mRNA (r=0.835,0.705,0.829,0.897,0.641,0.683,P ＜ 0.01,respectively).Conclusion Renal artery calcification may participate in and promote the progression of DN,and the BMP2 signal pathway may be an important regulating factor in DN with renal artery calcification.
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Objective To investigate the protective effect and mechanism of MST1 inhibition on kidney tissue in diabetic rats,and to find a new therapeutic target for diabetic nephropathy.Methods Total of 54 male SD rats enrolled in this study were divided into 3 groups including normal control (group A,n=18),MST1 inhibition group (Group B,n=18) and diabetes group (group C,n=18).Diabetes was induced by a single streptozotocin (STZ,50 mg/kg) injection in group B and group C.rats in group B received lentiviral vector contain Mst1 interference RNA (shRNA) and the rats in group C received empty vector.The end of 4th,8th and 12th week after modeling were considered as time points in this study.At each time point,the level of 24 hours urine protein (24-HUP),blood glucose and serum creatinine were examined.Pathological changes were observed with HE stain; Injury of podocyte and glomerular basement membrane (GBM) were examined with transmission electron microscope (TEM).The intensity and location of MST1 in kidney tissue were detected by immunohistochemistry.The level of MST1,Phosphorylated-MST1,nephrin,Caspase-3 and FasL were detected by western bloting.Results (1) At the starting point,there were no significant differences among groups in terms of weight,activity,eating and drinking.Since the end of 72nd hour after modeling,the levels of glucose in both group B and group C,compared to those in group A,significantly increased (P ＜ 0.05).There was no significant difference between group B and group C for glucose level at each time point (P ＞ 0.05); the level of 24-HUP increased significantly since the end of 4th week after modeling,and the level in group C was higher than its counterpart in group B at the same point (P ＜ 0.05); (2) There was no significant pathological lesion observed in group A.Without obvious K-W nodular changes,mesangial proliferation was observed in group B and group C.It was shown by TEM that podocyte fusion and thickening of the GBM could be found in group B and group C.The pathological change in group B was better than that in group C; (3) Compared to group A,it was shown by western blot that the levels of MST1,Phosphorylated-MST1,Caspase-3 and FasL in group B and group C were significantly higher (P ＜ 0.05),and the levels of nephrin in group B and group C were significantly lower (P ＜ 0.05) since the end of 4th week after modeling.Meanwhile,the levels of MST1,Phosphorylated-MST1,Caspase-3 and FasL in group B were significantly lower than that in group C at each time point (P ＜ 0.05),the level of nephrin in group B was significantly higher than the one in group C; (4) It was shown by immunohistochemistry that there was low MST1 expression in normal condition,especially in cytoplasm of tubular epithelial cells.The level of MST1 in group B and group C significantly increased after modeling,and the change could be the same as Western blot shown.Conclusions MST1 pathway could be involved in kidney injury induced by diabetes.MST1 inhibition could alleviate the kidney injury in STZ-induced diabetes animal model.
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To fully understand the characteristics of international students and importance, practicality and usefulness of internal medicine, many targeted measures were taken, including im-proving language level of teachers and students, applying case-based learning method and evidence-based medicine and strengthening the cultivation of humanistic spirit and professional ethics. Quality of internal medicine for international students was improved , which provided new ideas for clinical teaching reform.
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Objective To observe the change of intima/media thickness ratio and expression of inflammatory factors in renal small artery of diabetic rats, and to explore the correlations of intim/media ratio with inflammatory factors and vascular lesions of diabetic nephropathy (DN) rats. Methods Seventy healthy SD rats were randomly divided into diabetic nephropathy group (DN, n=40) and normal control group (N, n=30). DN rat model was induced by intraperitoneal injection of streptozotocin (STZ). Thirty-five DN rats were successfully established. N group received same dose of citrate buffer. Rats were sacrificed after 4, 12, 24 weeks respectively.The intima/media thickness ratio in renal small artery was detected by immunofluorescence. The monocyte chemoattractant protein-1 (MCP-1) protein and mRNA expression of renal small artery were detected by immunohistochemistry and in-situ hybridization at each time point. Results Blood glucose and urine protein excretion (24 h) at different time points in DN group were significantly higher than those of N group (P＜0.05). From the 12th week, Scr, BUN, serum phosphorus were significantly higher than those of N group (P＜0.05). At the 4th week, renal small artery had the expression of MCP-1 protein and mRNA. The expression increased gradually with time, reached the highest at the 24th week, and was significantly higher than that of N group at each time point (P＜0.05). Immunofluorescence results showed that as compared to N group, in the first 4 weeks, intima/media thickness ratio in DN group was not different, at the 12th week the ratio was higher but without significant difference, at the 24th week the ratio was significantly higher (P＜0.05). Small artery intima/media thickness ratio of DN group was positively correlated with MCP-1, cholesterol, triglyceride, serum phosphate (r=0.742, P＜0.01; r=0.740, P＜0.01; r=0.829, P＜0.01; r=0.580, P＜0.01). Conclusions The arterioles intima/media thickness ratio of early DN is significantly correlated with MCP-1, lipids and phosphorus. MCP-1 may be involved in the DN vascular disease.
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Objective To observe the expression of bone matrix proteins and the change of intima-tunica media thickness ratio in diabetic rat small renal artery and to explore their correlation and effects on diabetic nephropathy. Methods Seventy healthy SD rats were randomly divided into diabetic group(DN,n=40)and normal control group(N,n=30).DN rat model was induced by streptozotocin(STZ)intraperitoneal injection and the N group rats were given the same dose of citrate buffer.Thirty-five rats were successfully induced in DN group.The rats were sacrificed at week 4,12 and 24,respectively.The protein and mRNA expression of core-bind factor alpha 1(Cbfcd).bone morphogenetic protein 2(BMP-2)and matrix Gla protein(MGP)in smallrenal artery were detected by immunohistochemistry,in-situ hybridization and real-time PCR at each time point. Results Cbfctl and BMP-2 were expressed obviously in small renal artery of DN group by immunohistochemistry stain and in-situ hybridization from 4 to 24 weeks compared with N group at each time point,reaching the peak at week 24.Real-time PCR showed that the MGP mRNA was evidently increased at week 4,slightly decreased at week 12,lowest at week 24in DN group.The BMP-2 mRNA began to increase from week 4 onward,being peak at week 24in DN group.The ratio of intima to tunica media thickness had no significant difference in DN group compared with N group at week 4,but at week 12 and 24 there were significant difference between them.There was a positive correlation between Cbfα1 and BMP-2 expression,but they were negatively correlated with the expression of MGP.The ratio of intima to tunica media thickness was significantly-correlated with the expression of Cbfα1 and BMP-2. Conclusions The ratio of intima to tunica media thickness is positively correlated with Cbfα1 and BMP-2 in small renal artery of early DN.Cbfα1,BMP-2 and MGP may be involved in the progression of vascular lesions in DN.
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Objective To summarize the experience of prophylaxis and treatment of acute renal failure (ARF) following orthotopic liver transplantation (OLT). Methods The clinical data of 63 cases of ARF following OLT were analyzed retrospectively. Results Preoperatively, 12 out of 63 patients had renal dysfunction to varying degrees, 28 had serious peritoneal fluid and advanced hyperbil-irubinemia. Postoperatively, complication included pulmonary infection (28 cases), MOSF (26 cases) and intraperitoneal dropsy or empyema (9 cases). Cyclosporin A, mycophenolate and tacrolimus were all used to prevent rejection. Dopamine was used in some patients to improve renal perfusion. Meanwhile, diuretic and albumin, and fresh blood plasma were used to support patients. Twelve severe cases received CRRT. Average treatment duration was 50 h. Twenty-six patients died within one month postoperatively with the mortality rate of this group being 41.27 %. Conclusions The etiology of ARF following OLT is multifactorial. It's important to evaluate renal function preoperatively and to avoid infection, apply immunosuppressant individually and improve renal perfusion postoperatively.