Objective:To establish a mouse model of intra-jugular arteriovenous fistula (AVF) to screen differentially expressed genes in the process of intimal stenosis of AVF for investigating the abnormal expression signaling pathways and the mechanisms.Methods:Forty-six male C57BL/6 mice were randomly divided into AVF group ( n=23) and sham-operated group ( n=23). The AVF group underwent internal jugular arteriovenous fistuloplasty, and the sham-operated group separated the right external jugular vein and common carotid artery and then sutured the incision. The whole-genome sequences of mice with AVF stenosis were determined by transcriptomic reversible chain terminator and synthetic sequencing. The microarray data set was established, and the Benjamini & Hochberg method of gene microarray data analysis was applied to screen the differentially expressed genes. The differentially expressed genes were screened by R-language enrichment analysis. Then, gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) were performed. The subcellular localization of the differentially expressed genes was performed by BUSCA software. The protein network interaction of differentially expressed genes was analyzed by using STRING database and Cytoscape software. Results:In the AVF group, 21 mice were successfully modeled and 2 mice failed. Therefore, there were 21 mice in the AVF group and only 21 mice in the sham-operated group. This mouse internal jugular AVF model was innovated using the continuous-interrupted suture method, which improved the success rate of modeling this model. The differential gene sequencing analysis showed that there were 2 514 differentially expressed genes in the AVF process, including 1 323 up-regulated genes and 1 191 down-regulated genes. GO functional enrichment analysis showed that the differential genes were mainly enriched in metabolic process, activation, redox, mitochondria and so on. KEGG pathway enrichment analysis showed that the differential genes were enriched in metabolism, energy substance synthesis, diabetes, oxidative stress and so on. Statistical analysis of subcellular localization showed that the differences were mainly in mitochondrial proteins (24.24%), cytoplasmic proteins (17.51%), nuclear proteins (13.13%), cell membrane proteins (11.45%), and extracellular proteins (10.77%).Conclusions:Mitochondrial oxidative stress injury may be involved in the pathological damage process of endothelial proliferation stenosis in the AVF.

Objective:Clamping bilateral renal arteries with refined surgical methods to establish the rat renal ischemia-reperfusion injury (RIRI) model, and study the protective mechanism of ischemic preconditioning renal (IPC) tubular cell-derived exosomes in RIRI.Methods:25 female Sprague Dawley (SD) rats were divided into sham group, model group, inactivated group, normoxic group, IPC group. In the sham operation group, after bilateral renal arteries were dissociated, the back incision was disinfected and closed. The model group established RIRI model; RIRI models were established in inactivated group, normoxia group and IPC group, and then 200 μg of inactivated exosomes, normal exosomes and IPC exosomes were injected into the caudal vein 24 hours after operation. Serum creatinine (Scr) and urea nitrogen (BUN) levels were detected. The pathological changes of renal tissue were observed under light microscope. Transmission electron microscopy (TEM) was used to observe the shape and size of renal tubular exosomes. Nanoparticle tracking analysis (NTA)was used to detect the concentration and size of renal tubular exosomes.Results:Compared with the sham group, the Scr and BUN levels in the model group were significantly elevated ( P<0.01). Renal pathological changes in the model group showed damaged of the tubular structure, necrosis and shedding of tubular epithelial cells, and a large number of inflammatory cells accumulated in the renal interstitial tissue with varying degrees of edema. Compared with the inactivated group, the Scr and BUN levels significantly decreased in the normoxic group and IPC group ( P<0.01). Renal pathological changes in the normoxic group and IPC group showed that the renal tubular cell necrosis alleviated, inflammatory was reduced, the improved edema. Compared with the normoxic group, the Scr and BUN levels in the IPC group were further reduced ( P<0.01). Renal pathological changes in the IPC group showed that the inflammatory cells were significantly reduced, the cell edema was significantly improved, and the cell apoptosis was significantly reduced. Conclusions:Clamping bilateral renal arteries with refined surgical methods is the main and optimal way to build a rat model of RIRI. IPC tubular cell-derived exosomes have protective and repair effects on RIRI.

Overactive bladder (OAB) is the most bothersome symptom in lower urinary tract symptoms (LUTS). Current pharmacologic treatment aims to inhibit detrusor contraction; however, shows unsatisfied efficacy and high discontinuation rate. LIM kinases (LIMKs) promote smooth muscle contraction in the prostate; however, their function in the bladder smooth muscle remains unclear. Here, we studied effects of the LIMK inhibitors on bladder smooth muscle contraction and proliferation both

Objective:To investigate the effects and underlying mechanisms of phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT)/NF-κB signaling pathway in human kidney-2(HK-2) cells of hyperuricemic nephropathy.Methods:HK-2 cells were cultured in vitro and randomly divided into control group and experimental group. The experimental group was induced by high uric acid (720 μmol/L) immersion for 48 h to establish a cell model of hyperuricemic nephropathy in vitro and subsequently divided into hyperuricemic group, overexpressed protease activated receptor 2 (PAR2) and knockdown PAR2 group. The expressions of PAR2, PI3K, AKT, NF-κB mRNA were measured by real-time PCR. The expressions of PAR2, PI3K, AKT and NF-κB protein were measured by Western blotting. The expressions of tumor necrosis factor-α (TNF-α), monocyte chemotactic protein-1 (MCP-1), interleukin-6 (IL-6), pro-interleukin-1β (pro-IL-1β), interleukin-1β (IL-1β) and transforming growth factor-β1 (TGF-β1) were detected by enzyme linked immunosorbent assay (ELISA). Results:(1) Compared with the control group, the expressions of PAR2, PI3K, AKT and NF-κB mRNA and protein in hyperuricemic group were significantly increased (all P<0.05), the expressions of TNF-α, MCP-1, IL-6, pro-IL-1β, IL-1β and TGF-β1 in the supernatant in hyperuricemic group were significantly increased (all P<0.01). (2) Compared with the hyperuricemic group, the expressions of PAR2, PI3K, AKT and NF-κB mRNA and protein in overexpressed PAR2 group were significantly increased (all P<0.05), the expressions of TNF-α, MCP-1, IL-6, IL-1β and TGF-β1 in the supernatant were significantly increased (all P<0.05). (3) Compared with the hyperuricemic group, the expression of PAR2, PI3K, AKT and NF-κB mRNA and protein in knockdown PAR2 group were significantly decreased (all P<0.05), the expressions of IL-6, pro-IL-1β, IL-1β and TGF-β1 in the supernatant were significantly decreased (all P<0.05). Conclusions:In the process of uric acid-induced HK-2 cell damage, uric acid significantly up-regulates the expression of PI3K/AKT/NF-κB signaling pathway by activating PAR2, leading to a marked increase in inflammatory damage. Knocking down PAR2 inhibits the expression of PI3K/AKT/NF-κB signaling pathway, which can effectively reduce the inflammatory damage of HK-2 cells.

Objective:To assess the effect of stone and urine bacteria culture on the treatment of postoperative infection in percutaneous nephrolithotomy (PCNL).Methods:Between September 2016 and September 2018, 1060 patients with kidney stones treated with first-stage PCNL were included in the study. There were 614 male and 446 female patients, with the mean age (52.4±12.2) years. The mean stone burden was (1 499.6±1 435.3) mm 2. The midstream urine sample and the stone sample were sent for bacterial culture, identification of bacterial strain and antimicrobial susceptibility tests. The results of urine culture (UC), stone culture (SC) and their antimicrobial susceptibility, the details of perioperatively administered antibiotics and postoperative infections were recorded. The relationship between the postoperative infection and the SC was analyzed. Results:In 1 060 patients, 22 bacterial species were identified in UC and 52 bacterial species were identified in SC. The positive rate was higher in SC than in UC[31.8%(337/1 060)vs. 20.9%(222/1 060), P<0.001]. Escherichia coli was the most common bacteria in both UC and SC, but was more prevalent in UC than in SC [52.3%(116/222)vs. 43.6%(147/337), P<0.05]. E. coli cultured from UC and SC had high resistance to ampicillin, cefazolin, ceftriaxone, cefotaxime, levofloxacin, ciprofloxacin (all resistance rate >40%), but were sensitive to meropenem, cefoperazone/sulbactam, piperacillin/tazobactam, and amikacin (all resistance rate <10%). There was no statistical difference in the antibiotic resistance rates of E. coli from the UC and SC (all P >0.05). There were 111 (10.5%) patients who developed fever and 22 (2.1%) who developed urosepsis postoperatively. The incidences of postoperative fever and urosepsis were higher in the patients with positive SC than the patients with negative SC [23.7%(80/337)vs. 4.3%(31/723); 4.2%(14/337)vs. 1.1%(8/723), P<0.05]. Even in patients with negative UC, The incidence of postoperative fever was higher in the group with positive SC than the group with negative SC [17.9%(30/168) vs. 4.2%(28/670), P<0.05]. The incidence of postoperative fever in SC positive patients was lower if they were treated with sensitive antibiotics to the bacteria in stone than those treated with nonsensitive antibiotics [17.5%(22/126) vs. 27.5%(58/211), P<0.05]. Conclusions:The SC had high rate of culture positive, complicated bacterial species and high rate of multi-drug resistant. Positive SC was associated with increased incidence of postoperative infection even if the patients had negative UC. The SC might have a importance clinical value in the treatment of postoperative infection in PCNL.

Objective To assess and compare the incidence,clinical characteristics,treatment,and prognosis of acute heart failure patients from different grades hospitals in Beijing.Methods In this prospective internet prognosis registered study (Beijing AHF Registry),a total of 3 335 consecutive patients admitted to 14 emergency departments in Beijing from January 1st 2011 to September 23rd 2012 were enrolled.According to hospital grade,these patients were divided into two groups,349 patients were from secondary hospitals,and 2 956 patients were from tertiary hospitals.Results Among the 3 335 patients,the medium age was 71 (58,79) years,and male accounted for 53.16％.The most common underlying disease were coronary disease (43.27％),hypertension (17.73％),cardiomyopathy (16.07％) etc.The average treatment time in Emergency Department was 66.82 h.The emergency department mortality rate was 3.81％ (127 cases).The 30-day and 1-year cumulative all-cause mortality were 15.3％ and 32.27％,respectively.The 30-day and 1-year cumulative all-cause readmission were 15.64％ and 46.89％,respectively.Compared with patients in tertiary hospitals,patients in secondary hospitals had more onset acute heart failure patients (63.64％ vs.49.93％),shorter emergency department treatment time (12 h vs.41 h),lower discharge rate (3.43％ vs.37.45％) and emergency department mortality(1.58％ vs.4.09％).Compared with those in tertiary hospitals,1-year cumulative all-cause mortality (25.6％ vs.33.2％),cardiovascular disease mortality (20.2％ vs.26.0％),aggravated heart failure mortality (22.4％ vs.28.8％) were lower in secondary hospitals.Following propensity score matching,compared to tertiary hospitals,patients in secondary hospitals showed lower utilization rate of beta-blockers and ACEFARB (4.51％ vs.28.17％,1.41％ vs.9.58％),except the pironolactone.Conclusion Acute heart failure in emergency department is associated with a high mortality rate and readmission rate.There is still a big gap between guidelines recommend medication current treatments for acute heart failure.

Objective To investigate the effect and potential mechanism of autophagy inhibitor chloroquine on the calcium oxalate crystals formation in rats.Methods From September 2016 to October 2016,Thirty healthy male SD rats were randomly divided into 3 groups:control group,model group and chloroquine intervention group.The method to establish calcium oxalate stone model was drinking water with 1％ ethylene and 1％ ammonium chloride freely.The rats of chloroquine intervention group were treat with chloroquine (40mg/kg · d) by intraperitoneal injection.Modeling was finished after 28 days.The amounts of renalcalcium oxalate crystals were detected by polarizing microscope.For all groups,the amounts of autophagosome were detected by transmission electron microscope.Twenty four hour urine compositions for stone risk factors were detected.The expressions of oxidative stress injury related molecular markers (SOD,MCP-1 and 8-OHdG) and the expressions of autophagy markers (LC3 and P62) were detected by immunohistochemistry.The RNA expressions of SLC26A6 in kidney were detected by Real-time PCR.Results Compared to the model group,the amounts of renal calcium oxalate crystals were significantly reduced in chloroquine intervention group (32.37 ± 5.14 vs.4.18 ± 0.25,P ＜ 0.05).Compared to the control group,the level of autophagy was increased in the model group.Compared to the model group,the level of autophagy was inhibited in the chloroquine intervention group.For control group,model group and chloroquine intervention group,the excretion of urinary oxalate were (3.1 ± 1.5) mmol,(22.5 ± 8.1) mmol,(2.8 ± 1.2) mmol,respectively;the excretion of urinary citrate were (63.4 ± 7.4) mmol,(45.9 ± 9.5)mmol,(15.6 ± 8.2) mmol,respectively.Compared to the control group,the amounts of urinary oxalate weresignificantly elevated in model group (P ＜ 0.05),but citrate were significantly reduced in the chloroquineintervention group(P ＜ 0.05).For control group,model group and chloroquine intervention group,theexpressions of SOD were 42.24 ±4.16,19.21 ± 2.25,39.08 3.53,respectively;the expressions of MCP-1 were 4.02 0.51,8.45 ± 0.55,5.52 ± 0.34,respectively;the expressions of 8-OHdG were 7.16 ± 0.54,11.21 ± 1.12,8.67 ±0.34,respectively;the RNA expressions of SLC26A6 were 0.35 ±0.07,1.02 ±0.17,0.70 ± 0.06,respectively.Compared to the control group,the expressions of SOD were significantly reduced in the model group,but the expressions of MCP-1,8-OHdG and SLC26A6 were significantly elevated(P ＜0.05).Compared to the model group,the expressions of SOD were significantly elevated chloroquine intervention group (P ＜ 0.05),but the expressions of MCP-1,8-OHdG and SLC26A6 were significantly elevated(P ＜ 0.05).Conclusions The autophagy inhibitor chloroquine could inhibit the formation of calcium oxalate crystals induced by ethylene in rat kidney via inhibit the renal autophagy level and expressions of the SLC26A6,reducing the renal oxidative stress injury and urinary oxalate excretion.

Objective To identify risk factors of systemic inflammatory response syndrome (SIRS) after percutaneous nephrolithotomy (PCNL).Methods We retrospectively reviewed 438 renal calculi patients after PCNL from August 2015 to July 2016.Among them,there were 251 men and 187 women,the mean age was (49.4 ± 11.1) years.The positive preoperative urine WBC,culture and nitrite rates were 29.7％ (130),12.1％ (53) and 15.1％ (66),respectively.The stone size was (851.2 ± 663.6) mm2,the stone CT value was (960.4 ± 303.4) HU,the operative time was (63.5 ± 33.4) min,124 (28.3 ％) were infection stones and multiple-tracts PCNL was performed in 69 (15.8％) patients.Univariate and multivariate logistic regression analysis were used to analyze perioperative predictors after PCNL.Results Thirty-nine patients developed SIRS (8.9％) after PCNL.The univariate analysis showed that positive preoperative urine WBC,nitrite,culture,operation time,stone size and transfusion had significantly impacts on the outcome of postoperative SIRS after PCNL (P ＜ 0.05).Multivariable logistic analysis showed that positive preoperative urine nitrite (OR =5.990,P ＜ 0.001),stone size (OR =2.251,P =0.027) and transfusion (OR =7.501,P =0.007) were independently related to the postoperative SIRS.Conclusion The positive preoperative urine nitrite,stone size and transfusion are independent risk factors for postoperative SIRS after PCNL.

Objective To investigate the prevalence and awareness rate of adult diabetes mellitus and the risk factors in Liaoning Anshan. Methods A cross-sectional population survey was performed among residents in Liaoning Anshan and 2 500 subjects participated. Subjects whose fasting plasma glucose ≥ 5.6 mmol/L were confirmed by oral glucose tolerance test (OGTT). Results The standardized prevalence of adult diabetes mellitus over the age of 20 in Liaoning Anshan was 8.2%, and the standardized prevalence of pre-diabetes mellitus was 14.8%. The prevalence of diabetes mellitus increased with age (P<0.05). The awareness rate of diabetes mellitus in Liaoning Anshan was 52.3%(125/239), and the awareness rate of pre-diabetes mellitus was 8.7%(35/402). Logistic regression indicated that age, family history of diabetes, overweight or obesity, waist circumference, hypertriacylglycerolemia was the risk factors (P<0.05), while regular exercise was protective factor (P<0.05). Conclusions The current prevalence of diabetes mellitus and pre-diabetes mellitus in Liaoning Anshan is higher, and the awareness rate is lower.

Objective To investigate the protective effects ofDasangju Mixture on the liver injury induced by delayed-type hypersensitivity in mice; To discuss its mechanism of anti-inflammatory-immunity.Methods After enterocoelia intravenous injection of cyclophosphamide was used, 2,4,6-picryl chloride was used twice for exterior coating for skin of abdomen to cause sensation of skin. Then percutaneous transhepatic puncture was used to attack for modeling. Male Kunming mice were randomly divided into blank group, model group, high-, medium- and low-dose treatment groups. The high-, medium- and low-dose treatment groups were given Dasangju Mixture for gavage, once a day for 7 days. ELISA was applied to determine the serum levels of aspartate aminotransferase (AST), alamine aminotransferase (ALT) and tumor necrosis factor (TNF)-α. Pathological features of liver tissue were observed by HE staining.Results Compared with the blank group, the levels of AST, ALT and TNF-α in the model group increased significantly (P<0.05); Compared with the model group, the levels of AST, ALT and TNF-α in the high-, medium- and low-dose treatment groups decreased significantly (P<0.05,P<0.01,P<0.001); Compared with the model group, high-, medium- and low-dose treatment groups can better alleviate topical inflammation infiltration and edema of liver tissue.ConclusionDasangju Mixture has certain protective effects on the liver injury induced by delayed-type hypersensitivity in mice.