Objective:To evaluate the efficacy of homodimer neutrophil gelatinase- associated lipocalin (H-NGAL) rapid test kit in diagnosing peritoneal dialysis (PD)-associated peritonitis (PDAP).Methods:It was a multicenter prospective observational study. The PD patients from the nephrology clinics or wards at four hospitals: the First Affiliated Hospital of Nanchang University, the Third Hospital of Hebei Medical University, Taian Central Hospital, and Weifang People's Hospital from December 27, 2021, and July 18, 2022 were enrolled. The patients were categorized into PDAP and non-PDAP groups based on whether PDAP occurred at the time of enrollment. PD effluent samples were collected, and H-NGAL test was performed. The patients or their families used cassette-type, strip-type, and pen-type H-NGAL test kits to detect H-NGAL. Healthcare professionals only used the cassette-type H-NGAL kit to detect H-NGAL. All participants completed a questionnaire regarding the instructions for use of H-NGAL kit before testing, and a summary of patient experience and evaluation questionnaires after testing. The sensitivity, specificity, compliance rate, and 95% confidence interval ( CI) of H-NGAL rapid test kit for diagnosing PDAP were calculated using clinical diagnosis as the standard. Kappa test was used to assess the consistency between H-NGAL detection results and clinical diagnoses. Results:A total of 221 PD patients were enrolled, with 42 PDAP patients and 179 non-PDAP patients. The sensitivity and specificity of H-NGAL rapid test kit for diagnosing PDAP were 100% (95% CI 91.62%-100%) and 99.44% (95% CI 96.90%-99.90%), respectively, with accuracy rate of 99.55% (95% CI 97.48%-99.92%). The positive agreement rate was 97.67% (95% CI 87.94%-99.59%), and the negative agreement rate was 100% (95% CI 97.89%-100%). The consistency evaluation results showed that kappa value was 0.985 (95% CI 0.956-1.000). The results obtained by patients and their families using the H-NGAL rapid test kits were consistent across all three methods (cassette, strip, and pen-type H-NGAL test kits), and the results obtained by using the H-NGAL rapid test kits were also consistent between non-healthcare professionals and healthcare professionals. The kit instruction questionnaire and the experience and evaluation questionnaire showed that the overall satisfaction of patients with the H-NGAL rapid test kits was very good. Conclusions:H-NGAL rapid test kit exhibits high sensitivity and specificity in diagnosing PDAP and can serve as a rapid diagnostic method for PDAP.

Objective:To analyze the related factors affecting the use of autogenous arteriovenous fistula (AVF), and provide a theoretical basis for prolonging the service life of AVF in hemodialysis patients.Methods:This was a retrospective study. The patients undergoing AVF and using it to maintain hemodialysis (MHD) in the First Affiliated Hospital of Nanchang University from October 2004 to June 2017 were selected as study subjects to discuss the relevant factors affecting the service life of AVF. The data of general information, dialysis and laboratory examinations were collected through questionnaire surveys, hospital case system and hemodialysis record sheets. The patients were divided into the patency group and the dysfunction group according to the status of AVF, and the related factors were compared. Multivariate Cox proportional hazard model was used to analyze the influencing factors, and Kaplan-Meier survival curve was used to determine the service life of AVF, respectively.Results:A total of 187 subjects were included in the study. The patency group had 140 cases and the dysfunction group had 47 cases. There were statistically significant differences in the proportion of diabetes, the level of serum albumin, uric acid and parathyroid hormone (PTH) between the two groups (all P<0.05). Multivariate Cox proportional hazard regression analysis showed that diabetes ( HR=9.348, 95% CI 3.507-24.918, P<0.001) and hypoalbuminemia ( HR=12.650, 95% CI 2.925-54.714, P=0.001) were risk factors for the short service life of AVF. The results of Kaplan-Meier analysis showed that the service life of AVF in patients with diabetes was significantly shorter than that in MHD patients without diabetes (Log-rank χ2=13.191, P<0.001); the service life of AVF in patients with hypoalbuminemia was significantly shorter than that without hypoalbuminemia (Log-rank χ2=13.591, P<0.001). Conclusions:Diabetes mellitus and hypoalbuminemia are risk factors for the short service life of AVF. Therefore, intervention programs should be formulated to extend the service life of AVF.

Objective To explore the reasons for withdrawal from peritoneal dialysis (PD) in our hospital.Methods This was a single-center,retrospective cohort study.Patients who started PD in the Department of Nephrology,the First Affiliated Hospital of Nanchang University from November 1st,2005 to February 28th,2017,were enrolled,and followed up to May 31,2017.Patients who continued PD after May 31,2017 were as the control group.Patients who withdrew from PD were divided into 4 subgroups:death group,hemodialysis group,kidney transplantation group and loss of follow-up group.The clinical characters of 4 subgroups were compared with the control group.Results A total of 998 patients were enrolled with age of (49.36± 14.94) when PD started and median dialysis duration of 27.13(12.84,42.29) months,in whom 570 patients (57.11％) were male.Five hundred and seventeen dropout events were recorded,and the dropout rate was 51.80％.The main reason for withdrawal from PD was death (258 patients,49.90％),followed by hemodialysis (166patients,32.11％),kidney transplantation (66 patients,12.77％) and loss to follow-up (27 patients,5.22％).The leading cause of death was cardio-cerebro-vascular diseases (136 cases,52.71％),followed by infection (42 cases,16.28％),dyscrasia (20 cases,7.75％) and tumor (5 cases,1.94％).The main reason for transfering to hemodialysis was insufficient dialysis (76 cases,45.78％),followed by peritonitis (55 cases,33.13％) and catheter dysfunction (24 cases,14.46％).Compared with those in the control group,in the death group patients were older at PD commencement,and had higher proportions of hypertension,diabetes and cardio-cerebro-vascular diseases (all P ＜ 0.05).The proportions of male and diabetes mellitus were higher in the hemodialysis group than those in the control group (both P ＜0.05).Biochemical indicators showed that serum albumin and blood phosphorus were lower in the death group than those in the control group (both P ＜ 0.05);blood albumin was significantly lower in the hemodialysis group than that in the control group (P ＜ 0.05).Conclusions The main reasons for withdrawal from PD in our center are death and transfering to hemodialysis.The cardio-cerebro-vascular disease is the leading cause of death,and inadequate dialysis is the main reason for transfering to hemodialysis.

Objective To investigate the efficacy of leflunomide combined with prednisone in the induction therapy of proliferative lupus nephritis (LN).Methods A prospective,multicenter,randomized controlled clinical trial was conducted in patients with biopsy-proved proliferative lupus nephritis recruited from 15 renal centers from 2013 to 2015.Patients were randomized to two groups.Oral leflunomide or intravenous cyclophosphamide was given to patients in each group.Both groups received a tapering course of oral prednisone therapy.All patients were followed up for 24 weeks.The blood biochemistry,urine index,clinical curative effect and adverse reaction were recorded and analyzed statistically.Results A total of 100 patients were enrolled in this clinical trial,including 48 patients in leflunomide group and 52 patients in cyclophosphamide group.After 24 weeks,the overall response rate was 79％ (95％ CI 67％-90％) in the leflunomide group and 69％ (95％ CI 56％-82％) in the cyclophosphamide group.23％ (95％CI 11％-35％) of patients in leflunomide group showed complete remission compared with 27％ (95％CI 24％-30％) in cyclophosphamide group (P=0.35).The levels of 24-hr urine protein excretion,SLEDAI and anti-dsDNA antibody titers were decreased in patients treated with leflunomide group after 24-weeks treatment.And the levels of serum albumin and complement 3 after treatment were significantly higher compared with these before treatment.There was also no significant difference in changes of 24-hr urine protein excretion,SLEDAI score,anti-dsDNA antibody titers,serum albumin and complement C3 levels after treatment between two groups.Incidence of adverse events did not differ between the leflunomide and cyclophosphamide group.Conclusions Leflunomide combined with prednisone showed same efficacy compared with cyclophosphamide as induction therapy for lupus nephritis.Leflunomide might be an useful medicine in the induction therapy of lupus nephritis.

Objective To investigate the risk factors of all-cause mortality in diabetic patients on peritoneal dialysis (PD).Methods As a single-center retrospective cohort study,all incident PD patients who were catheterized at the First Affiliated Hospital of Nanchang University between November 1,2005 and February 28,2017 were included.Patients were divided into diabetes mellitus group (DM group) and non-diabetes mellitus group (NDM group).Outcomes were analyzed by Kaplan-Meier method.Multivariate Cox proportional hazards models were utilized to assess the risk factors of all-cause mortality.Results A total of 977 patients were enrolled.Compared with NDM group,patients in DM group were older (47.5±14.4 vs 59.3±11.3,P ＜ 0.01),had more cardiovascular disease (CVD) (7.5％ vs 20.3％,P ＜ 0.01),higher levels of serum hemoglobin (78.2±17.2 vs 82.3±14.6g/L,P ＜ 0.01),and lower levels of serum albumin (36.1±5.0 vs 32.7±5.6 g/L,P ＜ 0.01).The one-,three-and five-year patient survival rates of DM and NDM group were 89.7％,56.0％,31.9％ and 94.7％,81.3％,67.4％,respectively.Survival rate was significantly lower in DM group than in NDM group (x2=63.51,P ＜ 0.01).Stratified analysis showed that DM group had significant lower survival rate than NDM group in patients younger than 70 years old (x2=73.35,P ＜ 0.01),while survival rate was similar between the two groups patients older than 70 years old (x2=0.003,P=0.96).Multivariate Cox proportional hazards model analysis showed that DM (HR:1.74,95％CI:1.27-2.38,P ＜ 0.01),age (HR:1.05,95％CI:1.04-1.06,P ＜ 0.01),leukocyte (HR:1.06,95％CI:1.00-1.12,P=0.04) and triglyceride (HR:1.19,95％CI:1.07-1.32,P ＜ 0.01) were all independent risk factors for all-cause mortality of PD patients.However,age (HR:1.05,95％CI:1.04-1.07,P＜ 0.01) and alkaline phosphatase (HR:1.01,95％ CI:1.00-1.01,P=0.02) were independent risk factors for all-cause mortality of diabetic patients.Conclusions Long-term survival rate was lower in diabetic PD patients than in non-diabetic PD patients.DM,age,leukocyte and triglyceride were independent risk factors of mortality in PD patients.Age and alkaline phosphatase were independent risk factors of mortality in diabetic patients.

Objective To explore the risk factors and characteristics in patients with peritoneal dialysis who died in different periods.Methods The clinical data of new peritoneal dialysis patients in the Department of Nephrology and Peritoneal Dialysis Center of the First Affiliated Hospital of Nanchang University from November 1,2005 to February 28,2017 was retrospectively analyzed.The patients were divided into two groups according to the time of death:those who died within one year and died after one year.The risk factors of mortality between the two groups were analyzed by Cox regression model.Results A total of 997 patients were enrolled and 244 patients died.There were 69 patients (28.3％) died within one year and 175 patients (71.7％) died after one year.Cardiovascular and cerebrovascular disease was the dominating reason of death in both groups,accounting for 59.4％ (died within one year group) and 51.4％ (died after one year group) respectively.Cox regression analysis showed that for died within one year group,old age (HR=1.035,95％ CI:1.016-1.055,P＜ 0.001),low blood total calcium (HR=0.167,95％ CI:0.053-0.529,P=0.002),low albumin (HR=0.899,95％CI:0.856-0.943,P ＜ 0.001) and low apolipoprotein A1 (HR=0.274,95％CI:0.095-0.789,P=0.016) were risk factors associated with mortality.However,for died after one year group,old age (HR=1.053,95％CI:1.038-1.069,P ＜ 0.001),combined with diabetes (HR=2.181,95％CI:1.445-3.291,P ＜ 0.001) and hypertriglyceride (HR=l.204,95％CI:1.065-1.362,P=0.003) were risk factors associated with mortality.Conclusions The risk factors of mortality for peritoneal dialysis patients of different periods were not exactly the same.For died within one year patients,old age,low blood total calcium,low albumin and low apolipoprotein A1 were independent risk factors for mortality.However,for died after one year patients,old age,combined with diabetes,and high triglycerides were independent risk factors for mortality.

Objective To investigate the effect of suppressor of cytokine signaling 3 (SOCS3)on the proliferation of human mesangial cells stimulated by aggregated IgA1 (aIgA1) from patients with IgA nephropathy(IgAN),and explore its possible mechanism.Methods Serum monomeric IgA1 was isolated with jacalin affinity and Sephacryl S-200 HR chromatography from IgAN patients,and then heated to aggregated form (aIgA1).Human glomerular mesangial cells(HMC) were transfected with AdvSOCS3-IRES2-EGFP for 48 hours,and incubated with aIgA1 for 12-48 h.The cells were divided into blank control group,IgA1 group,IgA1 +Adv-EGFP group and IgA 1 +Adv-SOCS3-IRES2-EGFP group.The mesangial cell proliferation was observed through MTT,and the levels of SOCS3,TLR4,TGF-β1 protein and mRNA were detected through Western blotting and real-time PCR.Results HMC proliferation was promoted significantly after IgA1 stimulated at 24 h.Compared with control group,the protein and mRNA expression of SOCS3,TLR4,TGF-β1 were significantly increased in IgA1 group (P ＜ 0.05).Compared with IgA1 group and IgA1 +Adv-EGFP group,MTT absorbency was obviously reduced after incubation with aIgA1 for 24 h and 48 h in IgA+Adv-SOCS3-IRES2-EGFP group,and the protein and mRNA expression of TLR4 and TGF-β1 were significantly decreased in IgA1 +AdvSOCS3-EGFP group (P＜ 0.05).Conclusion Over-expression of SOCS3 may inhibit the proliferation of HMC stimulated by aIgA1,partly through down-regulating the expression of TLR4 and TGF-β1.

Objective To investigate the effect and mechanism of chitosan on vascular smooth muscle cell proliferation of uremia patients with arteriovenous fistula.Methods Primarily culturing the VSMCs of uremia patients with arteriovenous fistula and patients without uremia by explants adherent method,and taking the second generation.VSMCs from patients without uremia cultured with 20％ FBS medium were non-uremia group,VSMCs of uremia patients cultured with 20％ FBS medium were uremia group,VSMCs of uremia patients with 100 pg/ml chitosan were uremia+ chitosan group.The expression of α-SMA was detected by immunohistochemistry.The changes of migration and invasion of VSMCs were detected by scratches and transwell migration assays.The mRNA expressions of TLR4 and PCNA were measured by real-time PCR.VSMCs of uremia patients with arteriovenous fistula were intervened with different doses of chitosan (0,100 and 500 μg/ml),and the protein expressions of TLR4,MyD88 and NF-κB were detected by Western blotting.Results Compared with those in non-uremia group,in uremia group and uremia+chitosan group α-SMA was upregulated,migration and invasion of VSMCs were enhanced,and mRNA expressions of TLR4 and PCNA were increased (all P ＜ 0.05).Compared with those in uremia group,the level of α-SMA was significantly decreased,the ability of migration and invasion of VSMCs were decreased,and the mRNA expressions of TLR4 and PCNA were decreased (all P ＜ 0.05).TLR4,MyD88 and NF-κB protein expressions were reduced in concentration-dependent manner by 100 and 500 μg/ml chitosan.Conclusions (1) In vitro,chitosan decreases the ability of migration and invasion of VSMCs of uremia patients with arteriovenous fistula.(2) Chitosan inhibits the proliferation of VSMCs,which may be relevant in the decreased expressions of TLR4,MyD88 and NF-κB.

Objective To investigate the effects of angiotensin Ⅱ (Ang Ⅱ) or high glucose on the toll-like receptor 4 (TLR4) expression,inflammatory cytokines and fibrotic factors in human tubular epithelial cells (HK-2),revealing the innate immune-related pathogenesis of diabetic nephropathy (DN) which may have clinical implications.Methods Three TLR4 siRNA sequences were designed and synthetized.After transfection,the most effective siRNA was selected to use for further expriments.The experiment consisted of 2 parts.Part 1:Cells were divided into three groups:normal-glucose group (NG,5.5mmol/L glucose),mannose group (M,5.5 mmol/L glucose + 19.5 mmol/L mannose),High-glucose group (HG,25 mmol/L glucose),preliminary validated the effects of high glucose and high osmotic pressure.Part 2:Cells were divided into seven groups:NG group,HG group,Ang Ⅱ group,Ang Ⅱ + negative group,HG+ negative group,Ang Ⅱ + siRNA group and HG+ siRNA group.Real time PCR was used to analyze the mRNA expression of TLR4,myeloid differentiation factor 88 (MyD88),heat shock protein 47 (HSP47).Western blotting was used to observe the protein expression of TLR4,MyD88,HSP47,NF-κB,type Ⅳ collagen (ColⅣ).ELISA was used to detect the expression of monocyte chemotactic protein-1 (MCP-1) and interleukin-6 (IL-6).Results Compared with NG group,TLR4,MyD88,HSP47 mRNA and TLR4,MyD88,NF-κB,ColⅣ,HSP47 protein were highly expressed under high glucose or Ang Ⅱconditions (P ＜ 0.01),and the expression levels of MCP-1 and IL-6 also increased significantly (P ＜ 0.01).Compared with HG or Ang Ⅱ group,the above indicators were obviously inhibited in the TLR4 siRNA groups (P＜0.01).Comparison between blank vector transfected groups and HG group as well as Ang Ⅱ group indicated no statistic significance (P ＞ 0.05).Conclusions Both Ang Ⅱ and high glucose stimulate TLR4 expression,which result in the up-regulation of inflammatory and fibrotic factors in HK-2.Specific target of TLR4 gene silencing can block the TLR4 pathway that is activated by high glucose and Ang Ⅱ,and thus reduce the inflammatory and fibtogenic factors' release.TLR4 signal is the common innate immune response pathway which induces the release of inflammatory and fibrotic factors in HK-2 under high glucose or high angiotension conditions.

Objective To investigate the effete of chitosan on rabbit carotid artery internal jugular vein fistula intimal hyperplasia and its regulation on TLR4/NF-κB signaling.Methods A total of 28 New Zealand white rabbits were randomly divided into the control group(n=4),the model group(n=12) and the chitosan group(n=12).Model group and chitosan group rabbits were established respectively carotid artery internal jugular vein fistula models.After AVF surgery,chitosan was smeared on venous blood vessels and anastomosis.After 4,6 and 8 weeks,the rabbits were separately sacrificed and the AVF venous vascular tissues were taken.The pathological changes of AVF venous vascular tissue in each group were observed.The changes of α-SMA were detected by immunohistochemistry method.The mRNA expressions of PCNA and TLR4 in the tissues were measured by Real-time PCR.At the same time,the protein expressions of PCNA,TLR4,MyD88 and NF-κB were detected by Western blotting.The experimental data were processed by two-factor analysis of variance in statistics.Results (1) After 4 weeks,vascular intimal was thicked in mdel group.In intimal hyperplasia,α-SMA was staining,and then proliferation of vascular smooth muscle cell was significant.As time increasing,more intimal hyperplasia shown obviously,the expression of α-SMA significantly increased.Compared with model group,chitosan group significantly reduced the degree of intimal hyperplasia,the level of α-SMA was significantly decreased,vascular smooth muscle cell proliferation was also extraordinarily decreased.(2) Compared with control group,the expression levels of PCNA,TLR4,MyD88 and NF-κB increased with time.The indices of Chitosan group were markedly higher than control group,but significantly lower than model groups.Conclusion Chitosan can inhibit the proliferation of rabbit VSMCs.The mechanism may be concerned in down regulating TLR4-mediated signaling pathway,reducing the possibility of intimal hyperplasia of rabbit AVF venous blood vessels.