Objective To investigate the protective effect of wogonin on acute kidney injury(AKI)induced by li-popolysaccharide(LPS).Methods The model of septic-induced AKI was established on male C57BL/6J mice by a single intraperitoneal injection of LPS and normal C57BL/6J mice were used as normal control group.Twenty-four male C57BL/6 mice were randomly divided into 4 groups(6 mice in each group):normal control group(NC),normal control+wogonin(NC+WOG 12.5 mg/kg),LPS model group(LPS 10 mg/kg),LPS model+wogonin(LPS 10 mg/kg+WOG 12.5 mg/kg).After LPS intervention for 24 h,serum samples were collected to detect blood creatinine(CRE)and urea nitrogen(BUN)levels.HE staining and PAS staining were performed to observe the degree of renal pathological injury.Immunohistochemistry was performed to detect the degree of expression of inflammatory markers interleukin(IL)-1 β,IL-6 and tumor necrosis factor(TNF)-α in renal tissues.PCR was performed to detect the expression of KIM-1 and NGAL in renal tissues.Western blot was performed to detect the changes in protein expression of NF-κB signaling pathway subunits P65 and PP65 in renal tissues.Results Com-pared with NC group,CRE and BUN levels in LPS group increased(FCRE=60.90,P＜0.001,FBUN=82.13,P＜0.001);compared with LPS group,these indexes decreased in LPS+WOG group(P＜0.001).PCR test re-sults showed that compared with the NC group,the expression of KIM-1 and NGAL mRNA was significantly in-creased in LPS group(FKIM-1=146.3,P＜0.001,FNGAL=161.2,P＜0.001).In contrast,KIM-1 and NGAL mR-NA expression was decreased in the LPS+WOG group(P＜0.01).Renal histopathological examination showed that compared with the NC group,renal tissues of mice had renal tubular dilatation and inflammatory cell infiltration in LPS group;compared with LPS group,the number of tubular dilatation reduced and inflammatory cell infiltration was reduced in LPS+WOG group(FHE=721.4,P＜0.001;FPAS=518.9,P＜0.001).Immunohistochemical staining showed that the expression of IL-1 β,IL-6 and TNF-α was significantly increased in the LPS group;com-pared with the LPS groups(FIL-1β=114.6,FIL-6=108.9,FTNF α=251.6,all P＜0.001),these indexes de-creased in LPS+WOG group(all P＜0.01).Further studies using Western blot showed that the NF-κB signaling pathway of LPS-treated mice had been activated and produced a hyperphosphorylated state in comparison to the NC group(FPP65=13.02,P＜0.01),yet this pathway in the LPS+WOG group showed the opposite effect,namely attenuated activity and reduced phosphorylation when the control was LPS(P＜0.01).Conclusion WOG effec-tively blocked the NF-κB signaling pathway in the LPS-induced acute kidney injury model mice,thereby attenua-ting the inflammatory response and tissue damage in the kidneys of LPS-induced acute kidney injury mice.

Objective : To investigate the expression of Ca2 + -independent phospholipase A2 β (iPLA2 β) in human renal tubular epithelial cells (HK-2) induced by high glucose(HG) ，the relationship between iPLA2 β and ferroptosis and the protective mechanism of HG treated HK-2 cells． Methods: The HK-2 cells were treated with 30 mmol /L glucose，the overexpression model was constructed by transfection of iPLA2 β plasmid．Ferrostatin-1 ( Fer1) (an inhibitor of ferroptosis) and erastin (an activator of ferroptosis) were used as controls．After 36 hours of intervention，the kit detected the levels of superoxide (SOD) ，malonaldehyde(MDA) and iron in HK-2 cells．DCF immunofluorescence was used to detect intracellular reactive oxygen species ( ROS) ．The expression of ACSL4， GPX4，LPCAT3，TFR1 in HK-2 cells were measured by Western blot. Results : The expression of iPLA2 β downregulated in HG-induced injury of HK-2 cells．The levels of ROS and MDA in HK-2 cells increased，while the levels of GSH and SOD decreased．The expression of ACSL4，LPCAT3 and TFR1 decreased，and the expression of GPX4 increased in HK-2 cells．However，these indexes were improved after Fer-1 intervention．iPLA2 β overexpression could reduce the injury of HK-2 cells via attenuation of KIM-1． Further research revealed that iPLA2 β overexpression inhibited oxidative stress and ferroptosis in HK-2 cells injury induced by high glucose．Meanwhile， the improvement effect of iPLA2 β on HG-induced HK-2 cells damage could be eliminated by erastin． Conclusion iPLA2 β prevents HG-induced injury of HK-2 cells via regulating ferroptosis.

Objective : To study the correlation between cerebral white matter structure with cognitive function and blood biochemical indexes in patients with end stage renal disease (ESRD) . Methods : The diffusion tensor imaging (DTI) data of 64 ESRD patients and 47 age and sex matched healthy people were collected. Tract⁃based spatial statistics (TBSS) and XTRACT analysis methods were used to compare the differences in diffusion parameters between the two groups. Pearson correlation analysis was used to detect the correlation between various diffusion parameters and blood biochemical indexes and cognitive related scales. Results : The values of FA in the ESRD group generally decreased (P < 0. 05) . The values of MD, AD and RD obviously increased (P < 0. 05) . The scores of mini mental state examination (MMSE) and Montreal cognitive assessment (MoCA) decreased (P < 0. 01), while that of trail making test A part (TMT⁃A) increased (P < 0. 05) . In the ESRD group, the values of FA in the right anterior thalamic radiation, optic radiation, acoustic radiation, and cingulum were negatively correlated with the levels of creatinine and urea nitrogen, and positively correlated with MoCA′s scores, and the values of MD and RD of these tracts were positively correlated with the concentration of urea. The values of FA in optic radiation, acoustic radiation and left temporal of cingulum were negatively correlated with the scores of TMT⁃A. The values of FA in the vertical occipital fasciculus, inferior fronto⁃occipital fasciculus, left middle longitudinal fasciculus and forceps major were negatively correlated with the concentration of creatinine. The values of FA in the left arcuate fasciculus were positively correlated with the MoCA′s score, the values of MD and RD of these tracts were positively correlated with the concentration of creatinine, and negatively correlated with the scores of MMSE and MoCA. The values of FA in the right superior longitudinal fasciculus, the right corticospinal tract and the right frontal aslant tract werepositively correlated with the MoCA′s score, the values of MD and RD of these tracts were positively correlated with the concentration of urea; the values of MD and RD in the left fornix were positively correlated with the level of creatinine and urea. Conclusion The structural integrity of white matter in ESRD patients is extensively damaged, which is significantly associated with a variety of cognitive impairments. Serum creatinine and urea nitrogen may be risk factors for the changes in white matter.

Objective: To investigate the incidence and possible risk factors of small intestinal bacterial overgrowth(SIBO) in patients with chronic kidney disease(CKD). Methods : 92 patients with CKD were selected as subjects.SIBO was assessed by lactulose hydrogen methane breath test(LHMBT),fat tissue index(FTI) was determined using multifrequency bioelectrical impedance analysis, and gastrointestinal symptoms were evaluated by gastrointestinal symptom rating scale(GSRS).According to the results of LHMBT,the patients were divided into SIBO group and non-SIBO group.The demographic data, laboratory and clinical indicators, gastrointestinal symptoms and other differences between the two groups were compared.Binomial logistic stepwise regression was used to analyze the possible risk factors of SIBO in CKD patients. Results : The incidence of SIBO in these 92 patients was 47.8%.Compared with the non-SIBO group, the GSRS score of the SIBO group was higher, and the difference was statistically significant [(23.27±3.97)vs(21.13±4.39),t=2.451,P=0.016].Multivariate stepwise logistic regression analysis showed that low serum potassium(OR=0.396,95%CI0.176-0.893,P=0.025) and high FTI(OR=1.182,95%CI1.037-1.348,P=0.013) were independent risk factors for SIBO in patients with CKD. Conclusion The incidence of SIBO is high in CKD patients.Symptoms of dyspepsia are more prominent in SIBO positive patients.Low serum potassium and high FTI are independent risk factors for SIBO in patients with CKD.

Abstract: To investigate the role and mechanism of macrophage-to-myofibroblast transition(MMT) induced by exosomes from high glucose-treated renal tubular epithelial cells. Methods: Human renal tubular epithelial cells(HK2) were divided into glucose control group(5.5 mmol/L D-glucose) and high glucose group(30.0 mmol/L D-glucose) and cultured for 48 hours. The supernatant was collected by ultracentrifugation and identified by transmission electron microscopy and Western blot. PKH67 labelled exosomes were used to stimulate THP-1 macrophages. Laser scanning confocal microscopy was used to observe the phagocytosis process. The expression of inducible nitric oxide synthase(iNOS), α-SMA and CD206 was detected by flow cytometry and Western blot to determine the best concentration and time point. Laser scanning confocal microscopy was used to detect the fluorescence co-expression of rabbit polyclonal to mannose receptor(CD206), α-smooth muscle actin(α-SMA), collage type-Ⅳ(Col-IV) and fibronectin(FN). The expression of tumor growth factor-beta 1(TGF-β1), interleukin(IL)-6,IL-10 was separately detected by real-time quantitative PCR(qRT-PCR) and enzyme-linked immunosorbent assay(ELISA). The expression of TGF-β1, mothers against decapentaplegic homolog 3(Smad3), phosphorylated-smad 3(p-Smad3) in THP-1 macrophages was detected by Western blot. Results: The expression of cluster of differentiation 63(CD63)and tumor susceptibility gene 101 protein(TSG101)was positive, while the Calnexin protein was negative in the supernatant, confirming that the specimen was exosomes with high purity. THP-1 macrophages could internalize each group of exosomes. 40 mg/L exosomes for 96 hours were the best experimental condition. After being stimulated by high-glucose-exosomes, the percentage of M1 macrophages would climax in 24 hours, and the rate of M2 macrophages would climax in 96 hours. Immunofluorescence staining showed that exosomes released by HG-treated HK2 induced CD206, α-SMA, Col-IV and FN accumulation in cultured THP-1 macrophages. Compared with normal-glucose-exosomes, high-glucose-exosomes increased the expression of TGF-β1, IL-10 in M2 macrophages and decreased the expression of IL-6(allP<0.05). Moreover, TGF-β1 and p-Smad3 proteins expression also increased significantly(allP<0.05). Conclusion Exosomes secreted by renal tubular epithelial cells in high glucose environment can induce M2 macrophages transdifferentiate into myofibroblasts, and its mechanism may be related to activation of TGF-β1/Smad3 signal pathway.

Objective: To analyze Internet behaviors of children and adolescents and their influencing factors, and to provide scientific basis for health education guidance and prevention measures. Methods: A stratified cluster sampling method was used. From January to June 2019, 30 955 primary school students in 16 provinces (municipalities), from eastern, central, and western economic zones of China were selected to conduct a questionnaire survey, regarding general demographics, diet and exercise, tobacco, alcohol and sleep, Internet and electronics usage, etc. Results: Among them, 43.1% of students had never been online with girls (44.7%) higher than boys (41.8%)(χ2=21.04, P<0.01); 1.3% of students reported online time for more than 4 hours a day, boys were significantly higher than girls(χ2=15.87, P<0.01); Internet addiction was detected in 2.4% of elementary school students, with boys (3.0%) higher than girls(1.7%)(χ2=331.77, P<0.01), and the trend increases with grade. With sixth grade (3.3%) were significantly higher than children from the first grade (0.8%) and other grades(χ2=163.96, P<0.01). Children’s tendency to Internet addiction was 11.9%, of which boys were 13.0% and girls were 10.6%, boys were significantly higher than girls(χ2=117.66, P<0.01). Pearson correlation analysis revealed that, after controlling gender age, boarding school, family relationship, single parents and regrouped families, smoking, drinking sugary drinks, eating fried foods, and left-behind were positively related to Intenet addiction(r=0.06, 0.07, 0.27, 0.44, 0.07, 0.11, 0.09, P<0.01), while medium and high intensity sports, effective physical education, health education in school, AIDS education and parental restrictions on TV and computer games, were negatively associated with internet addiction(r=-0.74, -0.65, -0.04, -0.05, -0.63, P<0.01). Conclusion The internet addiction of Chinese primary school students is common and increases by year. High-intensity, high-density sports, effective physical education, and health education could help prevent Internet addiction among children.

Objective To elucidate the mechanism underlying eletroacupuncture's (EA's) amply-documented analgesic effect.To observe its effect on the pain threshold and on the expression of P2X4 receptor and microglia activation in the spinal cords of rats with neuropathic pain.To demonstrate whether or not interfering with A1 receptors and P2X4 receptors at the same time could enhance the analgesic effect.Methods A total of 40 SpragueDawley rats weighing 150 to 180 g were randomly divided into a sham group,a CCI group,an EA group,a 2-chloroN(6)-cyclopentyladenosine (CCPA) group and a CCPA+EA group,each of 8.Chronic constriction injury (CCI)was induced successfully in the rats of all groups except the sham group.Five days later,EA and 20 μL injections of 0.1 mm/L CCPA were applied to the rat analogues of the Zusanli (ST36) and Yanglingquan (GB34) acupoints once a day for 15 days for the rats in the appropriate groups.The mechanical withdrawal threshold (MWT) and the thermal withdrawal latency (TWL) were measured before the CCI operation and 20 days afterward.L4-L6 spinal cord tissue was then resected and the fluorescence intensity of P2X4 and OX42 receptors was detected using double label immunohistochemical staining.The correlation between the mean fluorescence intensity and the pain threshold gap was analyzed.Results The average MWT and TWL of the CCI group were significantly lower than in the other four groups.The expression of P2X4 receptor and OX42 in the spinal cord increased significantly in the CCI group compared to the other four groups.There was significant correlation between the mean fluorescence intensity of P2X4 and OX42 receptors and the gap in pain threshold,with correlation coefficients of 0.907 and 0.717 respectively.Conclusion P2X4 receptor and microglia activation might be involved in the development of neuropathic pain.CCPA and EA can inhibit the activation of microglia and reduce the activity of P2X4 receptors.The interaction between A1 receptors and P2X4 receptors can strengthen the analgesic effect of EA.

Objective To investigate the role and mechanism of macrophage activation induced by exosomes from high glucose-treated renal tubular epithelial cells.Methods (1) The supernatant of renal tubular epithelial cells which were cultured in normal glucose control group (5.5 mmol/L D-glucose) or high glucose group (30.0 mmol/L D-glucose) for 48 h were collected and ultracentrifuged to harvest exosomes.Exosomes were identified by transmission electron microscope and Western blotting.(2) Exosomes were labeled with the green lipophilic fluorescent dye PKH67 and cultured with THP-1 macrophage to investigate whether HK2-derived exosomes could be internalized by THP-1 macrophage.Observing the morphology microscopically and detecting the chemotaxis function of THP-1 macrophages in Transwell chamber after co-cultured with exosomes.The expression of inducible nitric oxide synthase (iNOS),tumor necrosis factor-α (TNF-α),Interleukin-1β (IL-1β),monocyte chemoattractant protein-1 (MCP-1) in cells and supernatants were separately detected by quantitative Real-time PCR (qRT-PCR) and enzyme linked immunosorbent assay (ELISA),and the expression of p-c-Jun NH2-terminal kinase (p-JNK),mitogen-activated protein kinase p-p38 (p-p38MAPK) and nuclear factor κB p65 (NF-κB p65) in THP-1 macrophages were detected by Western blot.Results (1) Vesicles that harvested by ultracentrifugation ranged in size from 30 nm to 100 nm and expressed exosomal marker CD63,TSG101 but absence of calnexin which is a marker of endoplasmic reticulum,suggesting that the exosomes were not contaminated with cells.(2) Results from laser scanning confocal microscope showed that each group of exosomes can be internalized by THP-1 macrophages.Compared with normal glucose exosomes group,high glucose exosomes had increased the expression of iNOS,TNF-α,IL-1β and MCP-1 in THP-1 macrophages (all P ＜ 0.01),moreover,p-JNK,p-p38 MAPK and NF-κB p65 proteins level also increased significantly (all P ＜ 0.01).Conclusions Exosomes from high glucose-treated HK2 cells can induce THP-1 macrophage activation and functional changes through MAPK/NF-κB pathway.

Objective To observe the influence of spinal cord injury (SCI) in hypothalamus AMP-activated protein kinase (AMPK) expression,and explore the mechanism of SCI-induced type 2 diabetes or insulin resistance.Methods Eighteen SD rats were randomly divided into model group,High Fat/High Sucrose/High Cholesterol Diet group and SCI group (n=6).The animals in the model group and High Fat/High Sucrose/High Cholesterol Diet group underwent a T10 laminectomy without weight-drop injury,and SCI models were induced by MASCIS Impactor method.Blood glucose and glucose tolerance tests were performed before experiment and 8 weeks after the experiment.At 8 weeks,the levels of serum inflammatory factors,tumor necrosis factor (TNF)-α,interleukin (IL)-1β,and IL-6,were observed by ELISA;Western blotting was used to detect the expression of phosphorylated (p) -AMPK in the hypothalamus.Results Blood glucose levels in the rats of High Fat/High Sucrose/High Cholesterol Diet group and SCI group were significantly higher than that in the model group (P＜0.05).Blood glucose levels 15 and 60 min after SCI in rats of SCI group were significantly higher than those in the model group (P＜0.05);Blood glucose levels 15,30,60 and 120 min after SCI in rats of High Fat/High Sucrose/High Cholesterol Diet group were significantly higher than those in the model group (P＜0.05).As compared with those in the model group,the expression of p-AMPK in the hypothalamus were significantly decreased,and the TNF-α,IL-1β and IL-6 levels were statistically increased in rats of High Fat/High Sucrose/High Cholesterol Diet group and SCI group (P＜0.05).Conclusion SCI could inhibit AMPK expression in the hypothalamus,activate inflammatory signaling pathway and enhance chronic systemic inflammation,and therefore,insulin resistance is induced.

Objective To explore the titre of serum anti-phospholipase A2 receptor antibodies (anti-PLA2 R antibodies) detected by the enzyme-linked immunosorbent assay (ELISA) to provide a more specific serological index for clinical diagnosis and disease judgment of membranous nephropathy (MN).Methods Thirty-four cases of MN confirmed by kidney biopsy ,32 inpatients with autoimmune diseases ,nephrotic syndrome and renal insufficiency in the nephrology department of our hospital and 12 persons un-dergoing physical examination were selected.The serum was collected for detecting anti-PLA2 R antibodies level.Then its diagnostic performance for diagnosing MM was analyzed by combining with serum TP ,ALB ,IgG ,IgA ,IgM ,C3 and C4 indicators.Results The medians of anti-PLA2 R antibodies titres in the MN group ,disease control group and healthy control group were 22.1 ,2.0 ,2.0 RU/mL respectively ,which had statistical difference between the MN group and other two groups (P<0.05).Seventeen cases of anti-PLA2R antibodies positive were in the MN group(positive rate50% )and 17 cases were negative,the disease control group and healthy control group all were negative.Its specificity and positive predictive value were 100% ,TP ,ALB and IgG had statistical difference between the MN group with the disease control group and healthy control group (P<0.05);the relative coefficients of anti-PLA2 R antibodies with TP ,ALB and IgG ,IgA ,IgM ,C3 and C4 were in turn -0.382 ,-0.344 ,-0.502 ,-0.295 ,0.062 , 0.005 and 0.241 respectively ,anti-PLA2R antibodies were negatively correlated with TP ,ALB ,IgG and IgA(P<0.01) ,positively correlated with C4(P<0.05) and had no relation with IgM and C3(P>0.05).Conclusion Anti-PLA2 R antibodies have higher specificity for the diagnosis of MN and can serve as the necessary and specific serologic detection indicator in the patients unable to conduct renal biopsy.Quantitative detection helps to condition judgment.