Objective To analyze the weight score and clinical application of 2019 European League Against Rheumatism/American College of Rheumatology (EULAR/ACR) systemic lupus erythematosus (SLE) classification criteria in lupus nephritis patients. Methods Lupus nephritis patients with renal biopsy results who were admitted in the First Affiliated Hospital of Zhejiang University College of Medicine between January 2014 and December 2018 were enrolled retrospectively. According to whether these patients were treated with glucocorticoids and/or immunosuppressants at the time of renal biopsy, they were divided into untreated group and post-treatment group. The weight scores were compared between the two groups, and the relationship between each weight score and remission after treatment was analyzed. Taking no remission as the end event, Cox regression analysis was used to analyze the influence of each weighted integral on the end event. Results A total of 153 patients were enrolled, including 131 (85.6%) females. These were 70 (45.8%) patients in the untreated group and 83 (54.2%) patients in the post-treatment group. The patients in the untreated group had higher scores of fever (>38.3℃), blood system involvement, low complement and positive specific antibodies than those in post-treated group (all P<0.05). In a median follow-up of 34 (6-50) months, 99 patients (64.7%) achieved complete remission, 38 patients (24.8%) achieved partial remission and 16 patients (10.5%) had no remission. With no remission as the endpoint event, univariate Cox regression analysis showed that proliferative lupus nephritis (renal score of 10 points vs 8 points) and neuropsychiatric involvement were the risk factors (both P<0.05), while multivariate Cox regression analysis showed that neuropsychiatric involvement (HR=4.758, 95%CI 1.324-17.101, P=0.017) was an independent risk factor. Conclusion The weight scores of 2019 EULAR/ACR SLE classification diagnostic criteria have certain predictive value for remission of patients with lupus nephritis.

Objective To evaluate the effect of preoperative plasma D-dimer level on the risk of cardiac surgery-associated acute kidney injury (CSA-AKI). Methods The clinical data of patients who underwent cardiac surgery with cardiopulmonary bypass in the First Affiliated Hospital of Nanjing Medical University from January 2017 to December 2018 were collected retrospectively. All patients were distributed into two groups (normal D-dimer group with D-dimer level≤0.55 mg/L and elevated D-dimer group with D-dimer level>0.55 mg/L) according to the D-dimer threshold of 0.55 mg/L and the differences of clinical data between the two groups were compared. Kaplan-Meier survival analysis method was used to analyze the difference of the cumulative incidence of CSA-AKI between the two groups. Logistic regression analysis and restricted cubic splines analysis were used to analyze the association between serum D-dimer and the incidence of CSA-AKI. Results There were 871 patients enrolled in the study with 427 females (49.0%) and age of (56.6±12.3) years, including 215 patients (24.7%) with high D-dimer and 259 patients (29.7%) with CSA-AKI. Compared with the normal D-dimer group, patients with elevated D-dimer had higher baseline serum creatinine, proportion of chronic kidney disease stage 3, international normalized ratio, fibrinogen, proportion of receiving renal replacement therapy and incidence of CSA-AKI (all P<0.05). The prothrombin time, operation time, extracorporeal circulation time, aortic occlusion time and hospital stay in the elevated D-dimer group were longer (all P<0.05), and the preoperative estimated glomerular filtration rate (eGFR) and hemoglobin levels were lower than those in the normal D-dimer group (both P<0.05). There was no statistical difference between the two groups in terms of age, gender, comorbid diseases, cardiac function classification, and hospital mortality (all P>0.05). Kaplan-Meier survival curve results showed that compared with the normal D-dimer group, the risk of CSA-AKI in the elevated D-dimer group was significantly increased (Log-rank χ²=14.227，P<0.001). The multivariate logistic regression showed that after adjusting variables including gender, age, diabetes mellitus, preoperative eGFR, cardiopulmonary bypass time and so on, the higher level of preoperative D-dimer (>0.55 mg/L) was still related to CSA-AKI (OR=1.476，95% CI 1.038-2.098，P=0.030). Restricted cubic splines analysis showed that the incidence of CSA-AKI raised when preoperative serum D-dimer concentration increased (non-linear P=0.262). Conclusion Patients with high preoperative serum D-dimer have an increased risk of CSA-AKI.

Objective To investigate the efficacy and safety of adrenocorticotropic hormone (ACTH) in treating primary nephrotic syndrome in children with dual resistance to glucocorticoids and calcineurin inhibitors (CNIs). Methods Clinical data of 6 children with primary nephrotic syndrome treated with ACTH in the Children's Hospital of Zhejiang University School of Medicine from January 1, 2015 to December 31, 2019 were retrospectively collected. All the enrolled patients were children with primary nephrotic syndrome with dual resistance to glucocorticoids and CNIs. All the 6 children were given 0.4-1.0 IU·kg^-1·d^-1 ACTH (total ≤25 IU)+5% glucose 500 ml intravenous infusion for 8 h during the hormone reduction process, with a course of treatment for 5 days, once a month, and continuous treatment for 3-6 months. Clinical data such as 24 h urinary protein quantification, serum albumin, serum cholesterol, estimated glomerular filtration rate (eGFR) level and glucocorticoid dosage were collected at equal time points at 6 months before treatment, at the beginning of treatment, at the end of treatment and at 6 months of follow-up after treatment of ACTH to evaluate the efficacy and adverse reactions. Results The onset age of 6 children was (4.89±1.77) years, and the age of the first treatment with ACTH was (9.49±3.06) years. All the 6 children completed 3 to 6 months of ACTH treatment, with 2 cases of complete remission, 2 cases of partial remission and 2 cases of no remission. At the end of ACTH treatment, 24 h urinary protein was significantly decreased (P=0.026), serum albumin level was significantly increased (P=0.003), and glucocorticoid dosage was significantly decreased (P<0.001) than before treatment. At 6 months after the end of ACTH treatment, there was no statistical significance in 24 h urinary protein, serum albumin and hormone dosage compared with the end of ACTH treatment (all P>0.05), and the blood cholesterol level continued to decrease (P=0.039). There was no significant change in eGFR during observation period (P>0.05). In the process of ACTH infusion, all the 6 children showed transient decrease in urine output, rash in 2 cases, and elevated blood glucose in 1 case, which could be spontaneously relieved after drug withdrawal. There were no serious cardiovascular events, renal impairment, infection and other adverse reactions. Conclusions ACTH has a good effect on children with primary nephrotic syndrome who are dual resistant to glucocorticoids and CNIs. ACTH can reduce proteinuria, decrease the dosage of glucocorticoids, improve the clinical remission rate, and has good security.

Objective To explore the mechanism of cisplatin-induced renal interstitial fibrosis and provide a new idea for the prevention and treatment of renal interstitial fibrosis. Methods Eight-week-old male C57BL/6 mice (specific pathogen-free) were used to carry out the experiment. The mice were divided into cisplatin group (10 mg/kg, n=6) and saline group (n=6) with intraperitoneal injection on day 0, 7 and 21, and sacrificed on day 28. The kidney tissues were collected for RNA Illumina high-throughput sequencing, real-time PCR, Western blotting, Masson staining and bioinformatics analysis. Results Through real-time PCR, Western blotting and Masson staining, a mouse model with cisplatin-induced renal interstitial fibrosis was successfully established. Through RNA Illumina high-throughput sequencing, 387 long noncoding RNA (lncRNA) and 2 427 mRNA were differently expressed between cisplatin group and saline group. The expression of the top two lncRNA was confirmed by real-time PCR with the same tendency as RNA sequencing. Complement C3 was found to be at the top among the different expressed mRNA by RNA sequencing. Several terms related to immunity were found to be within the top 20 terms through Gene Ontology (GO) enrichment analysis. Systemic lupus erythematous pathway (ko05322, Q=3.4E-17), including the complement cascade pathway, was found to be the top pathway through Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. The mRNA expression levels of C1q, C2, C3 and C4 were up-regulated remarkably in the cisplatin group by RNA sequencing than those in saline group (all P<0.05) and confirmed by real-time PCR. Conclusions Renal interstitial fibrosis can be induced by intraperitoneal injection of cisplatin periodically in mice, with complement cascade pathway activation in the diseased kidney.

Objective To investigate the effect of acteoside on the expressions of high mobility group box 1 (HMGB1) and nuclear factor-κB (NF-κB) in the renal tissue of diabetic nephropathy mice.Methods Among 20 healthy 8-week old C57BL/6J mice, 5 mice were randomly selected as normal control group, the rest were established as type 1 diabetes mellitus (T1DM) models by a single intraperitoneal injection of streptozocin (STZ, 150 mg/kg). T1DM mice were randomly divided into three groups: 5 mice without treatment, 5 mice treated with acteoside and 5 mice treated with irbesartan. After continuous administration for 8 weeks, serum, urine, and kidney tissue were collected for biochemical, pathological, and related mRNA and protein detection. The renal tubular epithelial cells (NRK-52E cells) were divided into control group (1 g/L glucose), high glucose group (4.5 g/L glucose) and high glucose+acteoside group (4.5 g/L glucose+32 μmol/L acteoside). Real-time PCR and Western blotting were used to assess the expressions of HMGB1 and NF-κB after 48 hours and 72 hours culturing. Results Compared with normal control group, blood glucose, 24-hour quantitative urinary protein, blood urea nitrogen (BUN), serum creatinine (Scr) and blood and urine HMGB1 were significantly increased in model group (all P<0.05), along with interstitial inflammatory cell infiltration and messangial matrix expantion, and the expressions of HMGB1 and NF-κB were significantly enhanced (all P<0.05). Compared with model group, histopathologic changes were alleviated and the mRNA and protein expression levels of HMGB1 and NF-κB were lower in the acteoside group (all P<0.05), while the blood glucose level was maintained at high level (P>0.05), excluding reduced quantitative 24-hour urinary protein, BUN, Scr, and serum and urine HMGB1 (all P<0.05). Compared with control group, the mRNA and protein expressions of HMGB1 and NF-κB were increased in high glucose group of NRK-52E cells (all P<0.05). Compared with high glucose group, the mRNA and protein expressions of HMGB1 and NF-κB in high glucose+acteoside group were down-regulated (all P<0.05). Conclusion Acteoside may alleviate the nephropathy in STZ-induced diabetic nephropathy mice by down-regulating the expressions of HMGB1 and NF-κB.

Objective To observe the expression of sirtuin 3 (Sirt3) and mitochondrial damage-associated proteins in lipopolysaccharide (LPS)-induced acute kidney injury mouse model and renal tubular epithelial cells, and to explore the role of Sirt3 in LPS-induced abnormal mitochondrial dynamics in renal tubular epithelial cells. Methods Eighteen specific pathogen free (SPF) male C57BL/6 mice were randomly assigned to control group, LPS 24 h group and LPS 48 h group. The control group was intraperitoneally injected with physiological saline (0.1 ml/10 g), and LPS 24 h group and LPS 48 h group were intraperitoneally injected with LPS (10 mg/kg) solution. Renal functional indexes of mice were analyzed by automatic biochemical analyzer. The pathological change of the kidney was observed by HE staining, and the expressions of dynamin-related protein-1 (Drp1), optic atrophy type 1 (Opa1) and Sirt3 were evaluated by Western blotting. Expression and distribution of Sirt3 in kidney was assessed by immunohistochemistry. Human renal tubular epithelial cells (HK-2) were exposed to 10 μg/ml LPS for 24 h，and the expression of Drp1, Opa1 and Sirt3 were detected by Western blotting. Cell apoptosis was assessed by Hoechst-33342 staining. After transfection to HK-2 cells with pcDNA3.1-Sirt3 recombinant plasmid, the expressions of Sirt3, Drp1, Opa1 and cell apoptosis were detected by the same methods as above. Results (1) The levels of blood urea nitrogen and serum creatinine in LPS group were significantly higher than those in control group (both P<0.05), and the pathological changes of kidney were obvious. (2) Compared with the control group, the expression of mitochondrial fission-associated protein Drp1 in renal tissue of LPS group was significantly higher (P<0.05), and the expression of mitochondrial fusion associated protein Opa1 was significantly lower (P<0.05). (3) Compared with the control group, the expression of Sirt3 in LPS group was significantly lower (P<0.05), and immunohistochemistry results showed that Sirt3 was mainly expressed in glomerular vascular endothelial cells and renal tubular epithelial cells. (4) In vitro, LPS stimulation induced increased Drp1 expression in HK-2 cells (P<0.05), decreased Opa1 and Sirt3 expression (both P<0.05), and increased apoptosis (P<0.05). (5) LPS-induced mitochondrial dynamics disturbance and apoptosis were alleviated by pcDNA3.1-Sirt3 recombinant plasmid transfection. Conclusions LPS can induce down-regulation of Sirt3 expression and disturbance of mitochondrial dynamics, and Sirt3 may play a protective role in LPS-induced acute kidney injury by regulating mitochondrial dynamics.