ABSTRACT
Objective:To screen the incidence of proteinuria in rural areas of Shanxi province and construct a risk prediction model of proteinuria based on machine learning algorithm.Methods:It was a cross-sectional investigation study. The residents ≥30 years old in rural areas of Shanxi province from April to November 2019 were screened by multi-stage stratified sampling method, and data from questionnaire surveys, physical examinations, and laboratory examinations were collected. Urine albumin/creatinine ratio ≥30 mg/g was defined as proteinuria, and the incidence of proteinuria was calculated. Subjects were divided into proteinuria group and non-proteinuria group. The machine learning binary classification model of proteinuria and non-proteinuria was constructed based on the stackable integrated logistic regression algorithm (SE-LR), logistic regression, support vector machine, decision tree, random forest and extreme gradient lift algorithms, respectively. The area under the receiver operating characteristic curve, accuracy, recall, and F1 weights were used to evaluate the predictive efficiency of the comparison models. Finally, the importance of the predictive features of the model with the best overall performance was ranked.Results:There were 8 869 rural residents included in the study, aged (58.59±9.49) years old, with 3 872 males (43.66%) and 4 997 females (56.34%). The prevalence of proteinuria in rural areas of Shanxi province was 13.49% (1 196/8 869). Blood pressure, pulse, body mass index, waist circumference, proportion of obesity or overweight, proportion of hypertension, proportion of moderate to severe salt intake, glycosylated hemoglobin, uric pH value, urinary specific gravity, proportion of positive urinary occult blood, proportion of positive urinary glucose, proportion of positive urinary ketone body, proportion of urinary red blood cell count ≥5/μl, proportion of urinary white blood cell count ≥10/μl and urinary α1 microglobulin in the proteinuria group were all higher than those in the non-proteinuria group (all P<0.05). The proportions of lack of exercise and drinking history in the proteinuria group were lower than those in non-proteinuria group (both P<0.05). The overall performance of SE-LR model was the best, with the area under the curve (0.736, 95% CI 0.719-0.746) slightly lower than that of the logistic regression model (0.745, 95% CI 0.680-0.762), and the highest accuracy (0.844), recall rate (0.621) and F1 weighting value (0.801). In the SE-LR model, the orders of importance of the top 10 features were urinary α1- microglobulin, urinary occult blood, urinary sugar, uric acid basicity, smoking history,overweight or obesity, body mass index, total cholesterol, glycosylated hemoglobin and hypertension. Conclusions:The prevalence of proteinuria is high in rural areas of Shanxi province. The risk prediction model of proteinuria established by machine learning algorithm can predict the risk of proteinuria and identify its risk factors, which can provide a scientific basis for disease prevention, intervention, and treatment in the community and clinic to a certain extent.
ABSTRACT
Acute kidney injury (AKI) is one of the most common diseases worldwide, and its incidence and mortality rate are increasing year by year. The key to improve the prognosis of AKI is early diagnosis and early intervention. As an endogenous nephrotoxic substance, the increase or decrease of serum calcium has a certain suggestive effect on the diagnosis and treatment of AKI. We reviewed the research progress of the relationship between serum calcium and AKI, to improve clinicians' understanding of the role of serum calcium in the diagnosis and treatment of AKI.
ABSTRACT
Objective:To explore the level of tibial growth plate chondrocyte mitophagy in young rats with chronic renal failure (CRF) and its effect on chondrocyte apoptosis.Methods:Male 4-week-old Sprague-Dawley rats were randomly divided into two groups according to random number table method: normal control group ( n=20, intragastric administration with distilled water) and CRF group ( n=20, given adenine suspension 150 mg·kg -1·d -1). All the young rats were sacrificed after continuous gavage for 6 weeks. The length of tibia was measured on X ray film, the width of tibia growth plate was measured and compared on histological section, and the apoptosis rate of chondrocytes in growth plate was detected by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) assay. The growth plate chondrocytes of two groups were isolated and cultured to the third generation in vitro, and the apoptosis rate of chondrocytes was detected by TUNEL assay. The co-localization of mitochondria and autophagy lysosomes in chondrocytes was observed by double fluorescence staining. Western blotting was used to detect the level of mitochondrial marker protein translocate of the outer mitochondrial membrane-20 (Tom-20) and autophagy marker light chain-3 protein (LC-3). The mitophagy of growth plate chondrocytes was observed by transmission electron microscope. Results:Compared with the normal control group, the tibia length of CRF group was shorter [(27.32±5.81) mm vs (35.43±3.61) mm, t=5.226, P<0.001], and the relative width of growth plate in histological section was narrower (0.56±0.19 vs 1.00±0.21, t=6.744, P<0.001). The apoptosis rate of chondrocytes in growth plate in CRF group was higher than that in the normal control group (17.2%±4.8% vs 5.1%±3.4%, t=6.505, P<0.001). The apoptosis rate of chondrocytes cultured in vitro in CRF group was higher than that in the normal control group (11.8%±6.2% vs 3.1%±1.2%, t=4.357, P<0.001). The result of double influorescence staining showed that there was co-localization between mitochondria and autophagy lysosomes in CRF group. Western blotting results showed that the levels of LC-3 protein ( t=8.944, P<0.001) and Tom-20 protein ( t=6.708, P<0.001) in CRF group were lower than those in the normal control group. Conclusion:The level of tibial growth plate chondrocyte mitophagy in young rats with CRF increases, which will lead to a decrease in the number of mitochondria, an increase in the apoptosis and a decrease in the number of chondrocytes, and eventually lead to dysplasia of tibia.
ABSTRACT
Objective:To prepare cisplatin-loaded anti-progastrin-releasing peptide (ProGRP) monoclonal antibody targeted nanobubbles, and to explore the proliferation inhibition effect and anti-cancer molecular mechanism of them on small cell lung cancer (SCLC).Methods:The cisplatin targeted nanobubbles were prepared by thin film hydrating method, and the physicochemical property were explored. The subcutaneous xenograft tumor models of SCLC in 10 nude mice were established, and the ultrasound molecular targeting development effect of cisplatin targeted nanobubbles was analyzed by using blank nanobubbles as control. Another 24 tumor-bearing nude mouse models were established and randomly divided into four groups: blank nanobubbles group, cisplatin group, cisplatin nanobubbles group, cisplatin targeted nanobubbles group. The tumor inhibition rate was calculated. The effect on SCLC proliferation was detected by CCK8 method. RT-PCR and Western blotting methods were used to detect SCLC proliferation related genes the P53, Rb, c-myc protein and mRNA expression level of change, the molecular regulatory mechanism was analyzed.Results:The cisplatin targeted nanobubbles were successfully prepared. The particle size was (467.3±42.3)nm, the structure was stable. The cisplatin targeted nanobubbles had a good effect of ultrasonic molecular development in SCLC xenograft.Compared with the control group, the proliferation of SCLC cells was significantly inhibited by cisplatin targeted nanobubbles. The RT-PCR and Western blotting analysis showed that compared with the control group, the mRNA and protein levels of the proliferation-related gene P53 and Rb in the cisplatin targeted nanovesicles group were significantly up-regulated, and the mRNA and protein levels of c-myc were significantly down-regulated (all P<0.05). Conclusions:The cisplatin targeted nanobubbles can inhibit the proliferation of SCLC, and may be used as a new potential targeted drug for the treatment of SCLC.
ABSTRACT
Objective:To evaluate the effect of edaravone on renal injury in rats with aldosterone-induced hypertension.Methods:Twenty-four clean-grade healthy male Sprague-Dawley rats, aged 6-8 weeks, weighing 200-220 g, were divided into 3 groups ( n=8 each) using a random number table method: sham operation group (S group), hypertension group (H group) and edaravone group (E group). The hypertension model was developed by subcutaneously embedding aldosterone osmotic pump (administration rate 0.75 μg·kg -1·h -1) for 4 weeks.After embedding osmotic pump subcutaneously, edaravone 10 mg/kg was injected via the tail vein every day for 4 consecutive weeks in E group, while normal saline 10 ml/kg was injected instead for 4 consecutive weeks in H group.BP-2010A noninvasive manometry device was used to measure the systolic pressure of tail artery before embedding osmotic pump and at 1, 2, 3 and 4 weeks after administration.After 4 weeks of administration, the 24 h urinary albumin concentration, plasma creatinine (Cr) and blood urea nitrogen (BUN) concentrations were measured, the bilateral kidneys were weighed, the right kidney weight/body weight ratio (RKW/BW) was calculated, the glomerular extramesangial matrix area/glomerular area ratio (M/G) was measured by PAS method, and the collagen volume fraction (CVF) was measured by Masson staining method, and the expression of aldosterone receptor (MCR) and type Ⅰ collagen in renal tissues was detected by Western blot. Results:Compared with group S, the systolic pressure was significantly increased, the concentrations of 24-h urinary albumin, plasma Cr and BUN were increased, the RKW/BW ratio, M/G and CVF were increased, and the expression of type Ⅰ collagen and MCR was up-regulated after embedding osmotic pump in group H ( P<0.05). Compared with group H, the systolic pressure was significantly decreased, the concentrations of 24-h urinary albumin, plasma Cr and BUN were decreased, the RKW/BW ratio, M/G and CVF were decreased, and the expression of type Ⅰ collagen and MCR was down-regulated after embedding osmotic pump in group E ( P<0.05). Conclusions:Edaravone can reduce renal injury in rats with aldosterone-induced hypertension, and the mechanism is related to down-regulation of MCR expression.
ABSTRACT
Objective:To explore the mechanism of highly expressed primary cilia in tibial growth plate chondrocytes accelerating chondrocytes differentiation in young rats with chronic renal failure (CRF).Methods:Forty male 4-week-old SD rats weighing (98±3) g were randomly divided into control group (intragastric administration with distilled water, n=20) and CRF group (intragastric administration with adenine suspension 150 mg·kg -1·d -1, n=20). All the young rats were sacrificed after continuous gavage for 6 weeks. The length of the growth plate was measured with histological sections. Immunofluorescence (IF) was used to detect the expression rate of primary cilia and the level of β-catenin, the key protein of Wnt/β-catenin signaling pathway in tibial growth plate chondrocytes. Chondrocytes isolated from growth plate in two groups were cultured in vitro to P3 generation, and the expression rate of primary cilia in chondrocytes, the levels of Indian hedgehog (IHH) and glycogen synthase kinase 3β (GSK3β) were detected by IF. Co-immunoprecipitation was used to detect the relationship between IHH and GSK3β. Results:Compared with the control group, the relative length of the growth plate was shorter in histological sections [(0.51±0.11) vs (1.00±0.08), t=16.11, P<0.001], the expression rate of primary cilia was higher [(26.3±5.5)% vs (7.6±1.9)%, t=14.37, P<0.001], and the level of β-catenin increased [(7.1±2.0) scores vs (3.6±1.0) scores, t=7.10, P<0.001] in CRF group. In vitro, the expression rate of primary cilia was higher in CRF group chondrocytes [(31.4±8.2)% vs (12.5±3.1)%, t=9.64, P<0.001] than that in control group. The level of IHH in CRF group increased than that in control group [(1 360±270) vs (310±84), t=16.61, P<0.001]. There was no significant difference in GSK3β level of chondrocytes between the two groups [(850±195) vs (780±140), t=1.30, P=0.200]. There was a direct interaction between IHH and GSK3β in CRF group chondrocytes. Conclusions:The expression levels of primary cilia and related protein IHH increase in tibial growth plate chondrocytes of CRF young rats. The IHH protein plays a direct interaction with GSK3β protein, Wnt/β-catenin signaling pathway antagonist, which leads to the activation of Wnt/β-catenin signaling pathway and final accelerated differentiation of chondrocytes. The rapid differentiation of chondrocytes causes the closing trend of growth plate.
ABSTRACT
Objective:To evaluate the relationship between edaravone-induced inhibition of pressure overload-induced myocardial remodeling and angiotensin Ⅱ type 1 receptor (AT1R)/mitogen activated protein kinases (MAPKs)/steroidogenic acute regulatory protein (StAR) signaling pathway in rats.Methods:Thirty-six clean-grade healthy male Sprague-Dawley rats, aged 2 months, weighing 200-220 g, were divided into 3 groups ( n=12 each) using a random number table method: sham operation group (S group), pressure overload group (POL group) and edaravone group (E group). The cardiac pressure overload was induced by ligation of thoracic aorta for 8 weeks.After the model preparation, 0.9% sodium chloride 10 ml/kg was intraperitoneally injected daily in group POL, and edaravone 10 mg/kg was given instead in group E for 8 consecutive weeks.After the model was successfully established, the left ventricular ejection fraction (EF) and ventricular shortening fraction (FS) were measured by two-dimensional ultrasound.The animals were sacrificed by bloodletting, and the heart weight/body weight ratio (HW/BW ratio) was calculated.Myocardial tissues were obtained for determination of the cross-sectional area (MSA) after HE staining, the collagen volume fraction (CVF) (using Masson′s staining), the expression of AT1R and StAR (by immunohistochemistry), extracellular signal-regulated kinase 1/2 (ERK1/2) and p38 MAPK phosphorylation levels (p-ERK1/2/ERK1/2 ratio and p-p38 MAPK/p38 MAPK ratio) (by Western blot) and the aldosterone content (by enzyme-linked immunosorbent assay). Results:Compared with group S, the HW/BW ratio, MSA and CVF were significantly increased, EF and FS were decreased, AT1R and StAR expression was up-regulated, and p-ERK1/2/ERK1/2 ratio, p-p38 MAPK/p38 MAPK ratio and aldosterone content were increased in group POL ( P<0.05). Compared with POL group, the HW/BW ratio, MSA and CVF were significantly decreased, EF and FS were increased, AT1R and StAR expression was down-regulated, and p-ERK1/2/ERK1/2 ratio, p-p38 MAPK/p38 MAPK ratio and aldosterone content were decreased in group E ( P<0.05). Conclusion:The mechanism of edaravone-induced inhibition of pressure overload-induced myocardial remodeling is probably associated with inhibiting the activation of AT1R/MAPKs/StAR signaling pathway in rats.
ABSTRACT
Objective:To investigate the role and mechanism of Nod-like receptor protein 3 (NLRP3) in chronic kidney disease (CKD)-related neointimal hyperplasia (NH) of vessels.Methods:Wild type C57BL/6J male mice were randomly divided into normal control group ( n=6) and experimental group ( n=18), by removal of 5/6 kidney and ligation of left common carotid artery to establish a NH model. After established successfully, the mice in NH experimental group were randomly divided into NH model group, NLRP3 inhibitor group, and drug control group ( n=6/group). C57BL/6J male mice with NLRP3 gene knockout group did not do any treatment after the establishment of NH model. After 3 weeks of feeding, the blood and vascular tissue samples of mice were collected. The pathological changes of vascular tissue samples in mice were observed by hematoxylin-eosin staining. The expressions and localization of NLRP3-related protein were observed by immunofluorescence staining. The expression of NLRP3 mRNA in vascular tissue was detected by quantitative real-time PCR. The activity of caspase-1 in vascular tissue was measured by colorimetric method. Human aortic smooth muscle cells (HASMCs) were treated with 10% uremic serum to simulate the body's internal environment during the uremic phase. NLRP3 small interfering RNA (siRNA) was transfected or NLRP3 inhibitor glibenclamide was added to the cell cultures. The expression of NLRP3 mRNA in HASMCs was detected by quantitative real-time PCR. The activity of caspase-1 in HASMCs was detected by colorimetric method. Results:Compared with the control group, the levels of serum creatinine and blood urea nitrogen were significantly increased in the NH model group (both P<0.01). The vascular histopathology showed that vascular intima thickened, vascular smooth muscle cells proliferated and hypertrophied, nuclei were deeply stained, and cells arranged disorderly and migrated to vascular intima in the experimental group. Quantitative analysis showed that the ratio of neointima to lumen increased significantly in the NH model group than that in control group ( P<0.01). Compared with the control group, the immunofluorescence staining of vascular tissue showed that the expressions of NLRP3, caspase-1, IL-18, IL-1β and proliferating cell nuclear antigen (PCNA) protein in the NH model group increased (all P<0.01), while the expression of α-SMA decreased ( P<0.01). NLRP3 was mainly located in vascular smooth muscle cells (VSMCs). VSMCs showed a synthetic phenotype. Compared with the NH model group, the expression of NLRP3, caspase-1, IL-18, IL-1β and PCNA protein in the NLRP3 inhibitor group and NLRP3 gene knockout group decreased (all P<0.01), the expression of α-SMA increased ( P<0.01), and the pathological changes of blood vessels alleviated. Compared with healthy serum group, the expression of NLRP3, IL-18, and IL-1β and bromodeoxyuridine (BrdU) uptake in uremic serum-stimulated group were increased (all P<0.01). After transfection of NLRP3 siRNA and addition of glibenclamide, the expression of NLRP3, IL-18, and IL-1β in VSMCs in uremic serum-stimulated group decreased, and BrdU intake decreased (all P<0.01). Conclusions:NLRP3 inflammatory bodies play an important role in promoting CKD-related neointimal hyperplasia of vessels, and glibenclamide can effectively reduce neointimal hyperplasia.
ABSTRACT
Objective:To explore the effect of Wnt/β-catenin signaling pathway on growth plate development of tibial growth plate in young chronic renal failure (CRF) rats.Methods:Four-week-old male SD rats were randomly divided into control group and CRF group ( n=20/per group). Control group was intragastric administration with distilled water, and CRF group was given adenine suspension (150 mg·kg -1·d -1). All the young rats were sacrificed after continuous gavages for 6 weeks. The full length of tibia was compared between the two groups. The width of tibia proximal growth plates was measured by micro-CT scanning, and the width of the growth plate was also measured in histological sections. Chondrocytes isolated from growth plate in two groups were cultured in vitro to P3 generation. Immunohistochemical staining was used to detect the expression of collagen Ⅱ, matrix metalloproteinase 13 (MMP-13) and β-catenin in chondrocytes. Western blotting was used to detect the protein expressions of collagen Ⅱ, MMP-13 and β-catenin. Results:Compared with the control group, the tibial length of rats in the CRF group was shorter [(27.32±5.81) mm vs (35.43±3.61) mm, t=5.226, P<0.001], the width of growth plate in micro-CT picture was more narrow [(0.72±0.22) mm vs (1.13±0.27) mm, t=5.096, P<0.001], and the relative width of the growth plate was also more narrow ( t=6.744, P<0.001) in histological sections. The results of immunohistochemistry and Western blotting showed the expressions of collagen Ⅱ in the CRF group decreased significantly ( t=8.212, P<0.001), MMP-13 ( t=13.091, P<0.001) and β-catenin ( t=7.534, P<0.001) increased significantly compared the control group in chondrocytes. Conclusion:The Wnt/β-catenin signaling pathway is highly expressed in the tibial growth plate of young rats with chronic renal failure, which leads to accelerated degeneration and differentiation of chondrocytes and a closure tendency of growth plate.
ABSTRACT
Objective:To understand the comprehensive geriatric assessment (CGA) scores in chronic kidney disease (CKD) patients aged 65 years and older, and analyze the related influencing factors of quality of life.Methods:A total of 189 patients who were over 65 years old and diagnosed with CKD in the Department of Nephrology of Shanxi Provincial People's Hospital from October 2016 to October 2019 were included retrospectively. The patients were divided into dialysis group ( n=90 cases) and non-dialysis group ( n=99 cases) according to whether dialysis or not. The concise CGA scores included age, basic activities of daily living (BADL), instrumental activities of daily living (IADL), and modified cumulative illness rating score for geriatrics (MCIRS-G). Pearson correlation analysis was used to analyze the relationship between different scale scores and clinical indexes. Multiple linear regression analysis was used to further analyze independent related factors of the quality of life in elderly CKD patients. Results:Compared with the non-dialysis group, the BADL score and IADL score in the dialysis group were significantly reduced [(70.00±33.28) vs (93.38±14.32), t=6.166, P<0.001; (9.78±7.12) vs (15.95±5.74), t=6.520, P<0.001], while the MCIRS-G score was significantly increased [(31.13±4.00) vs (27.29±5.17), t=-5.741, P<0.001]. Linear regression analysis performed on the data of non-dialysis group patients showed that estimated glomerular filtration rate (eGFR), serum uric acid (SUA), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), blood potassium and chlorine were positively correlated with BADL and IADL scores (all P<0.05). B-type natriuretic peptide (BNP) was negatively correlated with BADL score ( P<0.01). BNP and age were negatively correlated with IADL score (both P<0.05). Fasting blood glucose (FBG) was positively correlated with MCIRS-G or MCIRS-G other than kidney (both P<0.05), and eGFR, SUA, total cholesterol, and HDL-C were negatively correlated with MCIRS-G or MCIRS-G other than kidney (all P<0.05). Multiple linear regression analysis showed that eGFR was an independent influencing factor for BADL ( P<0.01). Age and eGFR were independent influencing factors for IADL (both P<0.05). Conclusions:The decline of quality of life in elderly CKD patients is related with eGFR, SUA, age, BNP and HDL-C levels, and eGFR and age are independent influencing factors.
ABSTRACT
Objective:To understand the laboratory characteristics for the diagnosis of immunoglobulin G4-related disease (IgG4-RD).Methods:The clinical data of 28 patients with IgG4-RD and renal damage (IgG4-RKD) diagnosed in our hospital from January 2017 to May 2019 were retrospectively analyzed. The correlation between serum IgG4 concentration and clinical features as well laboratory test results was analyzed. The 28 patients were divided into two groups: high serum IgG4 concentration group and normal serum IgG4 concentration group. The serum creatinine value, erythrocyte sedimentation rate, IgG concentration, IgA concentration, complement C3, C4 concentration, peripheral blood eosinophils, hemoglobin, IgG4/IgG and other related parameters were compared between the two groups. SPSS 20.0 statistical software was used for analysis. The two groups of measurement parameters were compared between groups by independent sample t test, non-normal measurement parameters were compared between groups by Mann-Whitney U test analysis, and the correlation between patients' IgG4 and each detection parameter was analyzed by Spearman correlation analysis. Results:Among the 28 patients, 17 were male and 11 were female, with an average age of (62±14) years. The serum IgG4 concentration increased in 75% of the patients ( n=21), with an average value of 3.01(1.41, 7.52) g/L, the serum IgG concentration increased in 64.3% of patients ( n=18), with an average value of 18.91 (12.88, 24.88) g/L, and the complement C3 decreased in 50% of the patients ( n=14), with an average value of(0.77±0.28) g/L. IgG4 was positively correlated with IgG ( r=0.422, P=0.025), IgG4/IgG ( r=0.951, P<0.01), ESR ( r=0.543, P<0.01) and peripheral blood eosinophils ( r=0.487, P<0.01), but negatively correlated with complement C3 ( r=-0.431, P=0.022) and C4 ( r=-0.504, P<0.01) levels. There were significant differences in IgG ( Z=-2.255, P=0.023), IgG4/IgG ( Z=-3.793, P<0.01), C3 ( t=7.380, P<0.01) and ESR ( t=-2.195, P=0.037) between the elevated IgG4 group and the normal group. Conclusion:Serological characteristics of IgG4-RKD combined with clinical manifestations may be able to diagnose IgG4-RKD in early stage.
ABSTRACT
Objective@#To investigate the effects and underlying mechanisms of aspirin on endoplasmic reticulum stress in podocytes induced by hyperlipemia.@*Methods@#Cultured podocytes were divided into four groups: control group, aspirin (100 μg/ml) group, oxidized low density lipoprotein (ox-LDL, 100 μg/ml) group, aspirin+ox-LDL group. The expression of protein kinase R-1ike endoplasmic reticulum kinase (PERK), eukaryotic translation initiation factor 2α (eIF2α), activating transcription factor-4 (ATF4) and CAAT/enhancer binding protein homologous protein (CHOP) at 6 h, 12 h, 24 h, 48 h were evaluated by real-time PCR. The related proteins of p-PERK and p-eIF2α at 24 h and ATF4 at 12 h were evaluated by Western blotting, respectively.@*Results@#The expressions of PERK, eIF2α peaked at 24 h, while ATF4 and CHOP peaked at 12 h in ox-LDL group and aspirin+ox-LDL group. Compared with control group, the expressions of PERK, eIF2α, ATF4 and CHOP were significantly higher in ox-LDL group at each times (all P<0.05). Compared with ox-LDL group, the expressions of the above indicators were significantly lower in aspirin+ox-LDL group at each times (all P<0.05). At 24 h, compared with control group, the expressions of p-PERK and p-eIF2α were significantly higher in ox-LDL group (both P<0.05). Compared with ox-LDL group, the expressions of p-PERK and p-eIF2α were significantly lower in aspirin+ox-LDL group (both P<0.05). At 12 h, the expression of ATF4 protein in each group was similar to that of mRNA. There were no significant difference in the expressions of all indicators between aspirin group and control group.@*Conclusions@#Hyperlipidemia may cause endoplasmic reticulum stress in podocytes by inducing phosphorylation of PERK and eIF2α, activating ATF4 transcription and inducing high expression of CHOP. Aspirin may partially block the PERK pathway, which may have protective effects for podocytes.
ABSTRACT
Objective:To investigate the effects and underlying mechanisms of aspirin on endoplasmic reticulum stress in podocytes induced by hyperlipemia.Methods:Cultured podocytes were divided into four groups: control group, aspirin (100 μg/ml) group, oxidized low density lipoprotein (ox-LDL, 100 μg/ml) group, aspirin+ox-LDL group. The expression of protein kinase R-1ike endoplasmic reticulum kinase (PERK), eukaryotic translation initiation factor 2α (eIF2α), activating transcription factor-4 (ATF4) and CAAT/enhancer binding protein homologous protein (CHOP) at 6 h, 12 h, 24 h, 48 h were evaluated by real-time PCR. The related proteins of p-PERK and p-eIF2α at 24 h and ATF4 at 12 h were evaluated by Western blotting, respectively.Results:The expressions of PERK, eIF2α peaked at 24 h, while ATF4 and CHOP peaked at 12 h in ox-LDL group and aspirin+ox-LDL group. Compared with control group, the expressions of PERK, eIF2α, ATF4 and CHOP were significantly higher in ox-LDL group at each times (all P<0.05). Compared with ox-LDL group, the expressions of the above indicators were significantly lower in aspirin+ox-LDL group at each times (all P<0.05). At 24 h, compared with control group, the expressions of p-PERK and p-eIF2α were significantly higher in ox-LDL group (both P<0.05). Compared with ox-LDL group, the expressions of p-PERK and p-eIF2α were significantly lower in aspirin+ox-LDL group (both P<0.05). At 12 h, the expression of ATF4 protein in each group was similar to that of mRNA. There were no significant difference in the expressions of all indicators between aspirin group and control group. Conclusions:Hyperlipidemia may cause endoplasmic reticulum stress in podocytes by inducing phosphorylation of PERK and eIF2α, activating ATF4 transcription and inducing high expression of CHOP. Aspirin may partially block the PERK pathway, which may have protective effects for podocytes.
ABSTRACT
Objective To investigate changes in intestinal bacteria in chronic renal failure (CRF),their diagnostic value for CRF,and correlations between specific bacterial genera and renal function.Methods Fecal specimens were collected from 56 patients with CRF and 38 healthy controls in the Nephrology Department and Medical Examination Center of Shanxi People's Hospital between August 2017 and January 2018.High-throughput sequencing analysis of 16S rDNA V3-V4 hypervariable regions was performed for intestinal bacteria.Intestinal bacteria in CRF patients and healthy subjects were analyzed for alpha,beta diversity,species composition analysis,and differential species analysis.The diagnostic value of the presence of specific intestinal bacteria for CRF was analyzed using a receiver operating characteristic curve (ROC).Pearson's correlation analysis was used to analyze the correlation between the presence of specific genera and the estimated glomerular filtration rate (eGFR).Results The alpha and beta diversity in the CRF group was different from that in the control group (P ＜ 0.05).At the phylum level,Verrucomicrobia were significantly less abundant in the CRF group than that in the control group (0.70％ vs 3.09％,P ＜ 0.001).The abundance of Actinobacteria was significantly greater in the CRF group than that in the control group (1.48％ vs 1.14％,P=0.036).At the genus level,the abundance of Akkermansia (0.96％ vs 3.90％),Parasutterella (0.47％ vs 0.93％),and Lactobacillus (0.07％ vs 0.48％) in the CRF group was significantly less than those in the control group (all P ＜ 0.01).The abundance of Alloprevotella (0.41％ vs 0.04％) and Clostridium Ⅳ (0.6％ vs 0.1％) was significantly greater than those in the control group (all P ＜ 0.05).The diagnostic value of CRF for the area under the ROC curve (AUC) for Akkermansia was 0.753,and that for Lactobacillus diagnostic value of CRF was 0.792.The combined AUC diagnostic value of CRF for detection of Akkermansia and Lactobacillus was 0.830,with high disease prediction value.Lactobacillus abundance was positively correlated with eGFR (R=0.29,P=0.029).Conclusions The diversity and structure of intestinal bacteria are altered in patients with CRF.The abundance of Akkermansia and Lactobacillus has diagnostic value for CRF.The abundance of Lactobacillus is positively correlated with eGFR.
ABSTRACT
Objective To investigate the expression of apoptosis stimulating protein two of p53 (ASPP2) in acute kidney injury (AKI) induced by carbon tetrachloride (CCl4) in mice. Methods Thirty-two male Balb/c mice were randomly divided into olive oil control group (control group, 8 mice) and CCl4 experimental group (experimental group, 24 mice). A mouse model of AKI was induced by a single high-dose abdominal injection of CCl4. The mice in the experimental group were sacrificed at 12 h, 24 h and 48 h after CCl4 injection. The mice in the control group were sacrificed 24 h after treatment. The serum and kidney tissue samples were collected. Serum biochemical indicators [serum creatinine (Scr) and urea nitrogen (BUN)] were measured by automatic biochemical analyzer. The pathological damage of kidney tissue was observed by hematoxylin-eosin (HE) staining and Periodate-Schiff (PAS) staining. ASPP2 positioning and expression level were observed by immunohistochemistry. The apoptosis of mouse kidney tissue was detected by in situ apoptosis. The expression of ASPP2 protein and ASPP2 mRNA in renal tissue were detected by Western blotting and quantitative real-time PCR. Results Compared with the control group, the levels of Scr and BUN were significantly increased in the experimental group (P＜0.01). Histopathology showed partial renal tubular brush margin detachment, renal tubular epithelial cell necrosis and nuclear disintegration in the experimental group. TUNEL staining showed that the apoptosis rate of renal tissue cells increased significantly in the experimental group (P＜0.01). Compared with the control group, the expression of ASPP2 in the experimental group increased at the early period and then decreased with the prolongation of injury time. The mRNA expression was consistent with the protein expression, and all reached the peak after 24 hours injury (P＜0.01). Immunohistochemistry showed that ASPP2 was mainly localized in the cytoplasm of renal tubular epithelial cells. Conclusions A single high-dose injection of CCl4 in the abdominal cavity can induce AKI in mice. The expression of ASPP2 is consistent with the degree of renal tissue damage. As the damage of renal tissue is aggravated, the expression of ASPP2 is gradually increased, which indicates that ASPP2 may be a damage factor.
ABSTRACT
Objective To observe the expression of parathyroid hormone - related peptide (PTHrp) receptor in tibial growth plate and its effects on tibial extension in chronic renal insufficiency rats. Methods Two-week-old male Sprague-Dawley rats were randomly divided into sham group, model group and enalapril group, each with 20 rats. In model group and enalapril group rats had chronic renal insufficiency induced by left ureteral obstruction, and rats were respectively given saline and enalapril by gavage after the operation. In sham group, left ureter was only exposed without ligation, and rats were given saline. The urine was collected 4 weeks after the operation and the total protein content was measured. Then all rats were killed. The concentrations of PTHrp, creatinine and urea nitrogen in intracardiac blood were detected. HE staining and Masson staining were performed on the left kidney to observe pathological changes of glomeruli and renal tubules. The total length of bilateral tibia was measured. The number of columnar cells in the growth plate proliferative zone was measured by safranin O staining and the expression of PTHrp receptor in the growth plate was detected by immunohistochemistry. Results The 24 h urine total protein, creatinine and urea nitrogen in model group were higher than those in sham group (all P＜0.05), while these 3 renal functional parameters in enalapril group were lower than those in model group (all P＜0.05). In model group and enalapril group rats had higher blood concentrations of PTHrp than that in sham group (all P＜0.05), but blood PTHrp in enalapril group was lower than that in model group (P＜0.05). HE staining and Masson staining showed that in the model group rats had severe tubular dilation, inflammatory cell infiltration and the tissue fibrosis, while in enalapril group renal tubules slightly dilated and had a few inflammatory cell infiltration and tissue fibrosis. Compared with those in the sham group, in model group the tibia length, the chondrocyte number of column structure in the growth plate proliferative zone and the PTHrp receptor decreased (all P＜0.05). But in enalapril group those indexes increased than model group (all P＜0.05). Conclusions Chronic renal insufficiency rats had increased PTHrp concentration in the blood but decreased PTHrp receptors expression in tibial growth plate, which lead to their limited tibial extension.
ABSTRACT
Objective To explore the relationship between intermedin (IMD) and renal interstitial capillary loss in IgA nephropathy (IgAN) patients.Methods Renal biopsy specimens collected from primary IgAN patients in our hospital (n=80) were compared with normal renal tissues.Expressions of IMD,CD31 and VE-cadherin were examined by immunohistochemical method,and plasma concentrations of IMD and TGF-β1 in 37 cases from the 80 cases were compared.The relationship between IMD and renal interstitial capillary loss in IgAN patients was analyzed.Results IMD and VE-cadherin in renal tubule interstitium expressions increased compared to the control group at the early stage of IgAN (P ＜ 0.05).CD31 expression remained unchanged at the early stage of pathological lesions of IgAN (P ＞ 0.05),but decreased at the early stage of clinical stage of IgAN compared to the control (P ＜ 0.05).Expressions of IMD,CD31 and VE-cadherin were reducing as the disease progressed,and the correlations of CD31 and VE-cadherin (r=0.517,P ＜ 0.01),IMD and CD31(r=0.655,P ＜ 0.01) or IMD and VE-cadherin (r=0.576,P ＜ 0.01) were positive.Plasma concentrations of IMD and TGF-β1 were higher than those of the control group at the early stage of IgAN (P ＜ 0.05),and the changes of IMD and TGF-β1were correlated positively (r=0.582,P ＜ 0.01).Conclusion Compared with the control group,expression of IMD in kidney tubules increases at the early stage of IgAN,and change of IMD correlates closely with the renal interstitial capillary loss.Plasma concentrations of IMD and TGF-β1 increase compared with the control group at the early stage of IgAN,and the changes of IMD and TGF-β1 are related closely.
ABSTRACT
Objective To investigate the glomerular microvascular injury and repair in patients with IgA nephropathy (IgAN) as well as its relationship with intermedin (IMD).Methods Eighty cases of renal tissue taken from patients first diagnosed as IgAN in Shanxi Provincial People's Hospital Affiliated to Shanxi Medical University and 15 cases of normal renal tissue were detected by the expression of glomerular IMD,CD31,and VE-cadherin through immunohistochemical method.ELISA method was used to detect VEGF and IMD of plasm from 31 normal subjects and 36 cases chosen from the IgAN patients.Their changes and internal relationship were analyzed according to Lee's and chronic kidney disease (CKD) classification.Results (1) Compared with the control group the expressions of CD31,IMD,and VE-cadherin in IgAN patients were statistically significant (P ＜0.01).Compared with the control group the levels of IMD and VEGF in plasma of IgAN patients in early stage of CKD group and late stage of CKD group were statistically significant (P ＜ 0.01).(2)Correlation analysis:the expression of glomerular CD31 and Lee's classification were negatively correlated (r=-0.232,P ＜ 0.05);glomerular IMD was negatively correlated with Lee's classification (r=-0.241,P＜0.05),while positively correlated with glomerular VE-cadherin (r=0.417,P＜ 0.01).VEGF in plasma of IgAN patients was positive correlated with CKD classification,BUN (r=0.458,0.409,P＜0.05),and negatively correlated with serum ALB (r=-0.532,P＜0.01).Conclusion Microvascular injury exists in patients with IgAN.The expression of VE-cadherin and IMD are positively correlated,suggesting that IMD may be involved in the progression of vascular protection and angiogenesis in IgAN.The contents of IMD and VEGF in plasma of IgAN patients increase,indicating that they may play a role in the progression of IgAN.
ABSTRACT
Objective To observe the changes of irisin in patients before and after hemodialysis (HD),as well as the differentiation of irisin change in patients with diabetes mellitus and protein energy waste.Methods Clinical parameters of patients on maintenance hemodialysis (MHD)in Shanxi People's Hospital from September 2016 to November 2016 were collected.A total of 33 cases were enrolled——14 cases of diabetic MHD group and 19 cases of non-diabetic MHD group as divided according to etiology.Based on the presence of protein energy waste,patients were also grouped into 17 cases with and 16 cases without protein waste.Before and after HD,the non parametric test was used to compare the changes of irisin in each group.Results After HD,the irisin value of 33patients with ERSD decreased,with the difference being statistically significant [0.666(0.218,1.365) ng/L vs 0.977(0.202,1.820) ng/L,P=0.01].The difference was not statistically significant in the diabetes MHD group;statistically significant in the non diabetes group [0.666(0.178,1.351) ng/L vs 0.913(0.100,1.497) ng/L,P ＜ 0.05];and not statistically significant in the protein energy group.The irisin of diabetic MHD group and non-diabetic MHD group were compared after HD:the difference was not statistically significant.Conclusions After HD,plasma irisin levels were reduced in patients with end-stage renal disease.Diabetes and protein wasting effects are not important for irisin at HD.
ABSTRACT
Deoxyglycychloxazol (TY501) is a glycyrrhetinic acid derivative which exhibits high anti-inflammatory activity and reduced pseudoaldosteronism compared to glycyrrhetinic acid. In this study, a sensitive and rapid liquid chromatography–tandem mass spectrometry (LC–MS/MS) method was established for the quantitation of TY501 in rat plasma. Plasma samples were treated by precipitating protein with methanol and supernatants were separated by a Symmetry C8 column with the mobile phase consisting of me-thanol and 10 mM ammonium formate (containing 0.1%of formic acid) (90:10, v/v). The selected reaction monitoring (SRM) transitions were performed at m/z 647.4-191.2 for TY501 and m/z 473.3-143.3 for astragaloside aglycone (IS) in the positive ion mode with atmospheric pressure chemical ionization (APCI) source. Calibration curve was linear over the concentration range of 5–5000 ng/mL. The lower limit of quantification was 5 ng/mL. The mean recovery was over 88%. The intra-and inter-day precisions were lower than 6.0% and 12.8%, respectively, and the accuracy was within 71.3%. TY501 was stable under usual storage conditions and handling procedure. The validated method has been successfully applied to a pharmacokinetic study after oral administration of TY501 to rats at a dosage of 10 mg/kg.