Objective:To investigate the protective effect and its mechanism of proscillaridin A (PSD-A) on podocyte injury in lupus nephritis (LN).Methods:Molecular docking and surface plasmon resonance techniques were used to analyze the binding status of PSD-A to signal transducer and activator of transcription 1 (STAT1). The immortalized human podocyte injury model in the lupus group was induced by the serum of systemic lupus erythematosus patients, and the control and PSD-A intervention (2 nmol/L, 4 nmol/L) groups were also set up. Six female 12-week-old C57BL/6 mice were designated as the control group, and 12 female 12-week-old MRL/lpr lupus mice were randomly divided into lupus group and PSD-A intervention group by random number table method. The PSD-A intervention group was intraperitoneally injected with 5 mg/kg PSD-A, once per week for 6 consecutive weeks. While the control group and the lupus group were intraperitoneally injected with the same volume of the solvent without PSD-A. Western blotting and real-time quantitative PCR were employed to detect the relative protein and mRNA expression levels of podocin, STAT1, and interferon-induced protein with tetratricopeptide repeat 1 (IFIT1) in podocytes of each group. Enzyme-linked immunosorbent assay was used to detect the levels of serum anti-double strand DNA antibody and interferon-α in mice. Coomassie brilliant blue was used to detect the urinary protein level. HE, PAS, Masson and PASM staining and transmission electron microscopy were used to observe the pathological changes of renal tissues. Immunohistochemistry was used to examine the protein expression of podocin, STAT1 and IFIT1 in renal tissues.Results:Molecular docking and surface plasmon resonance techniques proved that PSD-A could bind to STAT1 protein and they exhibited a robust binding affinity. The podocyte experiments showed that, compared with the lupus group, the relative expression levels of podocin protein and mRNA in the PSD-A intervention group were upregulated, while the relative expression levels of STAT1 and IFIT1 protein and mRNA were downregulated (all P<0.05). The animal experiments showed that, compared with the lupus group, the serum levels of anti-double strand DNA antibody, interferon-α, and urinary protein in PSD-A intervention group were decreased, the pathological damage of renal tissues was alleviated, and the injury of renal podocytes was reduced. Immunohistochemical staining showed that the relative protein expression levels of STAT1 and IFIT1 of renal tissues in the PSD-A intervention group were lower than those in the lupus group (all P<0.05). Conclusion:PSD-A can play a protective role in podocyte injury in LN, and its mechanism may be related to the inhibition of the STAT1 signaling pathway.

Objective:To investigate the protective effect and its mechanism of proscillaridin A (PSD-A) on podocyte injury in lupus nephritis (LN).Methods:Molecular docking and surface plasmon resonance techniques were used to analyze the binding status of PSD-A to signal transducer and activator of transcription 1 (STAT1). The immortalized human podocyte injury model in the lupus group was induced by the serum of systemic lupus erythematosus patients, and the control and PSD-A intervention (2 nmol/L, 4 nmol/L) groups were also set up. Six female 12-week-old C57BL/6 mice were designated as the control group, and 12 female 12-week-old MRL/lpr lupus mice were randomly divided into lupus group and PSD-A intervention group by random number table method. The PSD-A intervention group was intraperitoneally injected with 5 mg/kg PSD-A, once per week for 6 consecutive weeks. While the control group and the lupus group were intraperitoneally injected with the same volume of the solvent without PSD-A. Western blotting and real-time quantitative PCR were employed to detect the relative protein and mRNA expression levels of podocin, STAT1, and interferon-induced protein with tetratricopeptide repeat 1 (IFIT1) in podocytes of each group. Enzyme-linked immunosorbent assay was used to detect the levels of serum anti-double strand DNA antibody and interferon-α in mice. Coomassie brilliant blue was used to detect the urinary protein level. HE, PAS, Masson and PASM staining and transmission electron microscopy were used to observe the pathological changes of renal tissues. Immunohistochemistry was used to examine the protein expression of podocin, STAT1 and IFIT1 in renal tissues.Results:Molecular docking and surface plasmon resonance techniques proved that PSD-A could bind to STAT1 protein and they exhibited a robust binding affinity. The podocyte experiments showed that, compared with the lupus group, the relative expression levels of podocin protein and mRNA in the PSD-A intervention group were upregulated, while the relative expression levels of STAT1 and IFIT1 protein and mRNA were downregulated (all P<0.05). The animal experiments showed that, compared with the lupus group, the serum levels of anti-double strand DNA antibody, interferon-α, and urinary protein in PSD-A intervention group were decreased, the pathological damage of renal tissues was alleviated, and the injury of renal podocytes was reduced. Immunohistochemical staining showed that the relative protein expression levels of STAT1 and IFIT1 of renal tissues in the PSD-A intervention group were lower than those in the lupus group (all P<0.05). Conclusion:PSD-A can play a protective role in podocyte injury in LN, and its mechanism may be related to the inhibition of the STAT1 signaling pathway.

Objective:To investigate the correlation between Piwi-interacting RNA (piRNA) and diabetic nephropathy (DN).Methods:The differential expression profiles of piRNAs in renal tissues of patients with DN (experimental group) and renal tissues adjacent to tumors of patients with renal tumors (control group) were detected by high-throughput sequencing. The biological function of differentially expressed piRNAs was described by gene ontology and Kyoto encyclopedia of genes and genomes enrichment analysis. Real-time fluorescence quantitative PCR was used to detect the serum expression level of target piRNAs in patients with DN. Spearman correlation analysis was used to analyze the correlation between serum target piRNAs and clinical indexes of patients with DN.Results:The results of high throughput sequencing showed that there were 127 differentially expressed piRNAs between DN group and control group, with screening condition of |log 2(fold changes)|≥2 and P<0.05. Among them, there were 99 up-regulated piRNAs and 28 down-regulated piRNAs. The top 5 up-regulated piRNAs were piRNA-hsa-161686, piRNA-hsa-349255, piRNA-hsa-355720, piRNA-hsa-151229 and piRNA-hsa-154959, respectively. The top 5 down-regulated piRNAs were piRNA-hsa-1929960, piRNA-hsa-174194, piRNA-hsa- 148658, piRNA-hsa-172594 and piRNA-hsa-172421, respectively. The PCR verification results of 3 up-regulated genes and 3 down-regulated genes with low P values and high expression levels showed that serum expression level of piRNA-hsa-77976 was significantly down-regulated in patients with DN ( P=0.028), which was consistent with that of sequencing, while the expression levels of other genes were inconsistent with the sequencing results or had no statistical significance. Bioinformatics analysis results predicted that significantly differentially expressed piRNAs might participate in the regulation of DN through Rap1, Ras, PI3K-Akt and axon guiding pathways. The results of correlation analysis showed that the expression level of piRNA-hsa-77976 was negatively correlated with blood urea nitrogen ( r=-0.584, P=0.028), serum creatinine ( r=-0.637, P=0.014), cystatin C ( r=-0.738, P=0.003) and β2 microglobulin ( r=-0.822, P<0.001), and positively correlated with estimated glomerular filtration rate ( r=0.661, P=0.010). Conclusion:The differential expression of piRNA is closely related to DN, and may be used as a new biomarker for the diagnosis and prognosis of DN.

Vascular accesses are the lifelines for hemodialysis patients. This paper proposes to establish an integrative medicine with patient first model in the clinical practice of vascular access based on characters of hemodialysis patients and combined with the domestic and global innovation of management of vascular access. This model emphasizes the integration of vascular access plan with the whole life plan of renal replacement therapy, the combination of characters of vessel and entire condition of the patient, as well as balanced with patient's psychosocial characters. To implement this model in clinical practice, a multidisciplinary team with different professional background should be built, suitable job position should be set up and workflows should be formulated and optimized.

Objective:To investigate the risk factors for catheter-related bloodstream infection (CRBSI) in hemodialysis (HD) patients with tunnel-cuffed catheter (TCC) and construct a risk prediction model for the prevention and treatment of catheter infection.Methods:It was a retrospective study. Patients who had their TCC removed in Hemodialysis Access Center of the First Affiliated Hospital of Zhengzhou University from July to December 2020 were randomly divided into a training set (for model building) and a validation set (for model validation) in the ratio of 7∶3. The training set was divided into CRBSI group and non-CRBSI group with reference to the 2019 Kidney Disease Outcomes Quality Initiative clinical practice guidelines for vascular access, and the risk factors for the occurrence of CRBSI were analyzed. The odds ratio ( OR) values of the variables in the multivariate logistic regression analysis were applied to construct a risk prediction model, and the assessment ability of the model was validated in the validation set. Results:A total of 254 HD patients were included. The training set consisted of 179 patients with male-to-female ratio of 1.36∶1, age of (55.81±15.95) years old, median dialysis age of 18(8, 27) months, median TCC retention time of 15(5, 24) months, and 40 patients with confirmed CRBSI. Logistic regression analysis showed that, combined diabetes ( OR=2.711, 95% CI 1.174-6.258, P=0.019), history of catheter-related infection within 3 months ( OR=3.674, 95% CI 1.541-8.760, P=0.003), more than 4 times nursing interventions within 1 month ( OR=3.128, 95% CI 1.343-7.283, P=0.008), and central venous disease ( OR=2.572, 95% CI 1.130-5.854, P=0.024) were the independent influencing factors for CRBSI occurrence in HD patients with TCC. The OR values of the variables in the multivariate logistic regression were rounded to the assigned scores of the risk prediction model. The corresponding scores of each factor were summed in the training set to obtain the risk score. The receiver operating characteristic (ROC) curve was plotted, with area under the curve ( AUC) of 0.761(0.683-0.839) and maximum Youden index of 0.461, at which time the corresponding cut-off value was 6, with sensitivity of 90.0% and specificity of 56.1%. The model was validated in the validation set with AUC of 0.794(0.674-0.914) and cut-off value of 6, with sensitivity of 61.6% and specificity of 82.5%. Conclusions:Combined diabetes, history of catheter-related infection within 3 months, more than 4 times nursing interventions within 1 month, and central venous disease are the independent risk factors for CRBSI, and the prediction model based on the above factors has good efficacy in predicting the risk of CRBSI and can provide guidance for the prevention and treatment of CRBSI in HD patients.

Objective:To explore the influence of hypercholesterolemia on the risk of chronic kidney disease (CKD) in the middle-aged and elderly population and the gender differences.Methods:The data came from the "Epidemiological Survey of Chronic Kidney Disease among Adults in Urban Communities in Henan Province". The subjects came from 20 communities in Henan Province, aged ≥45 years old. Groups were based on the quartile of total blood cholesterol level and gender. Multivariate logistic regression and Cochran-Armitage trend test were used to analyze the effect of hypercholesterolemia on the risk of CKD and its gender differences.Results:A total of 4 779 subjects were enrolled into the study, with 1 934 males (40.5%) and 2 845 females (59.5%). The age was (61.3±7.7) years old and the blood cholesterol was (5.0±1.0) mmol/L. The prevalence rates of hypercholesterolemia, albuminuria, and reduced estimated glomerular filtration rate (eGFR) were 10.7%(305/2 845), 6.4%(182/2 845) and 2.8%(79/2 845) in females and 12.7%(245/1 934), 6.9%(133/1 934) and 2.3%(45/1 934) in males respectively. Compared with Q1 group, the prevalence of reduced eGFR in females were higher in Q2 and Q4 groups (both P<0.05). Among males, the prevalence of albuminuria and reduced eGFR increased with increasing blood cholesterol quartile (Cochran-Armitage trend test Z=12.231, 8.862, both P<0.001). Multivariate logistic regression analysis showed that hypercholesterolemia was an independent influencing factor for albuminuria and reduced eGFR ( OR=1.49, 95% CI 1.08-2.07, P=0.016 and OR=1.65, 95% CI 1.03-2.65, P=0.037, respectively). In subgroup analysis of different genders, female hypercholesterolemia was an independent influencing factor for albuminuria and reduced eGFR, while male hypercholesterolemia was not an independent influencing factor ( OR=1.54, 95% CI 0.96~2.46, P=0.075; OR=1.89, 95% CI 0.93-3.89, P=0.082, respectively). Further subgroup analysis based on the interquartile range of serum cholesterol levels found that female hypercholesterolemia was an independent influencing factor for reduced eGFR in the Q2 and Q4 groups ( OR=2.35, 95% CI 1.29-7.61, P=0.003; OR=2.51, 95% CI 1.38-8.39, P=0.001). In males, hypercholesterolemia was an independent influencing factor for albuminuria in the Q2, Q3 and Q4 groups ( OR=1.80, 95% CI 1.01-3.41, P=0.047; OR=1.85, 95% CI 1.02-3.35, P=0.044; OR=2.33, 95% CI 1.33-4.33, P=0.002). Conclusions:Hypercholesterolemia is an independent risk factor for CKD in middle-aged and elderly population, and there are gender differences, which provides a new idea for clinical prevention and control of CKD.

Objective:To analyze the influencing factors of prognosis of patients with diabetic kidney disease (DKD) in intensive care unit (ICU), and analyze their predictive value.Methods:Based on the inpatient information of more than 50 000 patients from June 2001 to October 2012 in the latest version of American Intensive Care Medical Information Database (MIMIC-Ⅲ v1.4), the data of DKD patients were screened out, including gender, age, body weight, comorbidities [hypertension, coronary heart disease, chronic obstructive pulmonary disease (COPD), chronic kidney disease (CKD)], sequential organ failure assessment (SOFA) score, the length of ICU stay, the incidence of mechanical ventilation, vasoactive drugs and renal replacement therapy during the ICU hospitalization, complications of other diseases [ventilator-associated pneumonia (VAP), urinary tract infection (UTI), diabetic ketoacidosis (DKA), acute myocardial infarction (AKI)] and prognosis of ICU. At the same time, the blood routine and biochemical data of the first 24 hours in ICU and the extremum values during the ICU hospitalization were collected. Multivariate Logistic regression analysis was used to screen the prognostic factors of DKD patients in ICU, and receiver operating characteristic (ROC) curve was plotted to analyze the predictive value of death risk factors.Results:416 DKD patients were screened out, 20 patients were excluded due to data missing, and finally 396 patients were enrolled, including 220 survival patients and 176 dead patients. Compared with the survival group, the patients in the death group were older (years old: 57.13±13.04 vs. 52.61±14.15), with lower rates of hypertension and CKD (11.4% vs. 23.6%, 26.7% vs. 41.4%), higher SOFA scores and baseline values of blood urea nitrogen (BUN), serum creatinine (SCr) and blood K + [SOFA score: 5.86±2.79 vs. 4.49±2.56, BUN (mmol/L): 18.4±10.0 vs. 14.8±9.0, SCr (μmol/L): 387.2±382.8 vs. 284.6±244.9, K + (mmol/L): 4.64±0.99 vs. 4.33±0.86], and longer ICU stay [days: 2.65 (1.48, 5.21) vs. 2.00 (1.00, 4.00)], and the differences were statistically significant (all P < 0.01). Further analysis of laboratory tests extremum values during ICU hospitalization showed that the maximum (max) and minimum (min) values of white blood cell (WBC), BUN and SCr, and K +max in the death group were significantly higher than those in the survival group [WBC max (×10 9/L): 17.3±10.3 vs. 14.5±7.3, WBC min (×10 9/L): 7.9±4.1 vs. 6.7±2.7, BUN max (mmol/L): 23.8±10.4 vs. 18.8±10.2, BUN min (mmol/L): 11.0±6.6 vs. 9.3±6.6, SCr max (μmol/L): 459.7±392.5 vs. 350.1±294.4, SCr min (μmol/L): 246.6±180.3 vs. 206.9±195.4, K +max (mmol/L): 5.35±0.93 vs. 5.09±0.99], and the minimum values of hemoglobin (Hb min) and glucose (Glu min) were significantly lower than those in the survival group [Hb min (g/L): 87.4±14.5 vs. 90.6±16.5, Glu min (mmol/L): 4.0±1.7 vs. 4.6±2.0], and the differences were statistically significant (all P < 0.05). The incidences of mechanical ventilation and vasoactive drugs during ICU hospitalization in the death group were significantly higher than those in the survival group (37.5% vs. 24.1%, 32.4% vs. 20.0%, both P < 0.01), and the incidences of UTI and AMI in the death group were significantly higher than those in the survival group (29.5% vs. 19.1%, 8.5% vs. 3.6%, both P < 0.05). Multivariate Logistic regression analysis showed that age [odds ratio ( OR) = 1.019, 95% confidence interval (95% CI) was 1.003-1.036, P = 0.023], SOFA score ( OR = 1.142, 95% CI was 1.105-1.246, P = 0.003), WBC min ( OR = 1.134, 95% CI was 1.054-1.221, P = 0.001) and BUN max ( OR = 1.010, 95% CI was 1.002-1.018, P = 0.018) were risk factors of death of DKD patients in ICU. ROC curve analysis showed that the area under ROC curve (AUC) of combination of risks factors of death was 0.706, the sensitivity was 61.6%, and the specificity was 73.2%. Conclusions:In order to prevent DKD patients from getting worse in ICU, we should pay close attention to the blood biochemical indexes, especially the renal function indexes, and give timely treatment. At the same time, we should actively prevent the occurrence of complications such as infection and cardiovascular disease.

Objective:To compare the effect of various antigen retrieval methods in paraffin sections of renal biopsy tissue, and explore the best antigen retrieval method.Methods:Forty-five renal biopsy specimens were collected from the First Affiliated Hospital of Zhengzhou University, including lupus nephritis ( n=10), membranous nephropathy ( n=10), IgA nephropathy ( n=10) and amyloidosis glomerulopathy ( n=15). Five retrieval methods (including high pressure thermal retrieval combined with trypsin retrieval, microwave thermal retrieval combined with trypsin retrieval, high pressure thermal retrieval, microwave thermal retrieval and gastroprotease retrieval) were used for immunofluorescence staining of paraffin sections. Renal tissue specimens were divided into six groups according to different antigen retrieval methods, frozen section specimens used as a control group. The immunofluorescence semi-quantitative scores of paraffin sections of the five heat-repairing antigen methods and frozen sections were compared. Results:Immunofluorescence staining of hyperbaric thermal retrieval combined with trypsin retrieval group and microwave thermal retrieval combined with trypsin retrieval group were similar with those of frozen sections. Compared with the control group, there were no significant difference in the semi-quantitative immunofluorescence scores between the two groups (all P>0.05). However, Immunofluorescence staining of hyperbaric thermal retrieval, microwave thermal retrieval, pepsin digestion had significantly higher false negative rate than those of frozen sections. Compared with the control group, the difference in semi-quantitative immunofluorescence score was statistically significant (all P<0.05). Conclusion:High pressure heat retrieval combined with trypsin retrieval or microwave heat retrieval combined with trypsin retrieval is the first choice of antigen retrieval methods.

Objective:To investigate the expression level of vitamin D receptor (VDR) in podocytes of diabetic kidney disease (DKD) and its role in podocyte injury and proteinuria.Methods:(1) Sixty-five patients who had been diagnosed with type 2 diabetes mellitus (with or without albuminuria) were enrolled in this study and 25 age-and sex-matched healthy control subjects were enrolled. According to the ratio of urinary excretion of albumin/creatinine (ACR), the type 2 diabetes mellitus patients were classified into without proteinuria group (ACR<30 mg/g, n=24), microalbuminuria group (ACR 30-300 mg/g, n=18) and clinical proteinuria group (ACR>300 mg/g, n=23). Another 25 patients with DKD confirmed by renal biopsy were selected as the DKD group. Normal kidney tissue samples were taken from the same period of urinary surgical department for 10 cases of renal tumors in patients with renal resection. The test indicators in each group were compared. The VDR expression in blood, urine samples and kidney tissues of patients was detected by real-time quantitative PCR, ELISA and immunohistochemistry, and then were compared among different groups. The correlation between VDR and ACR was analyzed by Pearson correlation analysis. (2) Male db/db mice with genetic background of C57BLKs/J and db/m mice born in littermates were randomly divided into normal control group (group A), DKD control group (group B), DKD treated with dimethyl sulfoxide group (group C), DKD treated with paricalcitol (VDR agonist) group (group D). The C and D groups were treated by continuous intraperitoneal injection for 8 weeks, and the group A and B were not treated. The mice were started to intervene continuously for 8 weeks at the age of 10 weeks. At 22 weeks of age (12 weeks after starting intervention), the biochemical indexes of the mice's body weight, blood and urine were compared. The changes of β-catenin and VDR were detected by Western blotting. The expressions of podocyte marker protein podocin and podocyte injury protein α-SMA were observed by immunofluorescence. Results:(1) Compared with the normal healthy control group, the plasma levels of VDR mRNA and protein in diabetic patients without proteinuria, microalbuminuria and clinical proteinuria were lower (all P<0.05). Compared with the diabetic patients without proteinuria, the plasma levels of VDR mRNA and protein in diabetic patients with microalbuminuria and clinical proteinuria were lower (all P<0.05). (2) Compared with the normal healthy control group, the plasma levels of VDR mRNA and protein in diabetic patients without proteinuria and DKD patients were lower (all P<0.05). Compared with diabetic patients without proteinuria, the plasma levels of VDR mRNA and protein in the DKD group were also lower (both P<0.05). (3) Immunohistochemical results showed that the expression of VDR in kidney tissue of DKD group was significantly lower than that of normal control group. (4) The relative level of VDR mRNA in plasma of patients with DKD was negatively correlated with ACR ( r=-0.342, P<0.05). (5) The levels of VDR in urine supernatant of each group showed opposite trends with the plasma levels. (6) Western blotting results showed that the expression of β-catenin protein in groups B and C was higher than that in group D (both P<0.05), and the expression of VDR protein was lower than that in group D (both P<0.05). Immunofluorescence results showed that the expression of podocin in groups B and C was lower than that in group D (both P<0.05), and the expression of α-SMA was higher than that in group D (both P<0.05). Conclusion:VDR overexpression relieves podocyte injury and proteinuria in DKD.

Diabetes kidney disease (DKD) is a serious renal complication of diabetes mellitus, also the main cause of end-stage renal disease (ESRD). Early diagnosis and treatment are very important. Mass spectrometry (MS) can detect biomarkers in blood, urine, tissue and other samples of DKD patients, which has become a research hotspot, and its great significance in early diagnosis, pathogenic research, treatment effect and prognostic evaluation.