ABSTRACT
Objective:To explore the expression of type 2 complement receptor (CR2) in mesangial cells of the renal tissue in IgA nephropathy (IgAN) and its possible mechanism involved in complement C3 deposition.Methods:The demographic data, samples of plasma and renal tissues of primary IgAN patients diagnosed by renal biopsy in the Guangdong Provincial People's Hospital from August 2021 to May 2022 were collected. According to the fluorescent intensity of mesangial complement C3 deposition, the patients were divided into complement C3 deposition ≥2+ group and complement C3 deposition <2+ group. The circulating IgA and complement C3 levels were detected by enzyme linked immunosorbent assay (ELISA). The influencing factors of kidney prognosis, plasma IgA and complement C3 levels were compared between the two groups. Immunofluorescence was used to detect the expression of IgA, complement C3 and CR2 in the renal mesangial cells of IgAN patients and normal renal tissues around renal carcinoma. Human mesangial cells were cultured in vitro and randomly divided into control group and experimental group. The experimental group was incubated with IgA protein (2 g/L) for 8 hours. The expressions of CR2 protein and mRNA were measured by Western blotting and real-time fluorescence quantitative PCR. The biological function of differential genes was analyzed by gene ontology (GO) and Kyto encyclopedia of genes and genomes (KEGG) enrichment analysis. Results:A total of 75 patients with IgAN were included in this study, including 50 patients in the complement C3 deposition ≥2+ group and 25 patients in the complement C3 deposition <2+ group. The proportions of patients with urine red blood cell count negative, 1+, 2+ and 3+-4+ in the complement C3 deposition ≥2+ group were 2.0%, 8.0%, 18.0% 72.0%, respectively, which were more serious than those in the complement C3 deposition <2+ group (4.0%, 4.0%, 52.0%, 40.0%) ( Z=-2.320, P=0.020). Meanwhile, the proportion of S1 in Oxford pathological classification in the complement C3 deposition ≥2+ group was higher than that in the complement C3 deposition <2+ group (68.0% vs. 40.0%, χ2=5.389, P=0.020), and there were no statistically significant differences in gender, age, 24-hour urinary protein, serum creatinine, other indicators of Oxford pathological classification between the two groups. ELISA results showed that plasma IgA concentration in the complement C3 deposition ≥2+ group was higher than that in the complement C3 deposition <2+ group [3.62 (2.95, 5.53) g/L vs. 2.72 (2.15, 4.24) g/L, Z=2.405, P=0.016], and the plasma complement C3 concentration was lower than that in the complement C3 deposition <2+ group [199.6 (116.0, 328.0) mg/L vs. 319.2 (158.3, 454.5) mg/L, Z=-2.383, P=0.017]. Spearman correlation analysis showed that the complement C3 deposition intensity was positively correlated with IgA deposition intensity in mesangial area ( rs=0.441, P<0.001). Immunofluorescence results showed that there was colocalization of IgA and complement C3 in the glomeruli of IgAN patients. The expression of CR2 in the kidney was consistent with complement C3 deposition, and CR2 was colocalization with complement C3. In vitro experiments, the expression of CR2 in IgA protein group was higher than that in the control group ( P<0.05). GO and KEGG enrichment analysis found that IgA protein induced active changes in various pathways of mesangial cells. Conclusion:IgA protein induces mesangial cells to express CR2 and participates in complement C3 deposition, which may be an important mechanism of complement C3 activation in IgAN.
ABSTRACT
Objective To explore the relationship between clinical manifestations and pathological changes in diabetic nephropathy (DN) and to assess the predictive power of the pathologic classification for DN established by the Renal Pathology Society in 2010.Methods Patients with type 1 or type 2 diabetes and biopsy-proven DN in the Third Affiliated Hospital of Sun Yat-sen University between January 2004 to June 2014 were enrolled in the present study and were followed-up until 31 December 2014.The outcome was defined as renal end-points including renal replacement therapy and doubling of serum creatinine as well as all-cause mortality.The laboratory and histologic data were analyzed and outcomes were assessed using survival analysis.Results Fifty-seven people enrolled in this study were categorized into Class Ⅱa (n=9),Class Ⅱb (n=9),Class Ⅲ (n=25) and Class Ⅳ (n=14) while no participants belonged to Class Ⅰ.The changes of Class Ⅱa were slight and those of Class Ⅳ were severe both in the clinical data (diabetic duration,blood pressure,estimated glomerular filtration rate,urine protein excretion rate,albumin and hemoglobin) and the pathological data (percentage of global glomerulosclerosis,percentage and scoring of interstitial fibrosis and tubular atrophy,scoring of interstitial inflammation and incidence of large vessel lesions).There were no significant differences between Class Ⅱb and Ⅲ in the above variables except for the scoring of arteriosclerosis.The mean follow-up duration was 25.9 months.Twenty-five patients (43.9％) reached the renal outcomes and six people (10.5％) reached all-cause mortality.The survival analysis showed that there were significant differences among the renal survival curves of different glomerular classes and of different interstitial and vascular scorings,but not in the survival curves related to all-cause mortality.Conclusions The glomerular classes are not completely associated with renal prognosis.The clinical manifestations and renal outcomes are benign in Class Ⅱa,moderate but similar in Class Ⅱb and Ⅲ and severe in Class Ⅳ.The glomerular classification and interstitial and vascular scorings are associated to renal prognosis while their associations with mortality remain to be verified.
ABSTRACT
Objective To analyze the related factors of micro-albuminuria and macroalbuminuria in type 1 diabetes mellitus (DM) in the classification tree model,and to screen the high risk population of diabetic kidney disease.Methods 394 patients with type 1 diabetes were enrolled in the hospital from 2008 to 2015.According to glomerular filtration rates and urine albumin quantification,the patients were divided into type 1 diabetes group (299 cases),micro-albuminuria group (73 cases) and macro-albuminuria group (22 cases).The classification tree model was utilized to analyze related factors of the different stages of proteinuria,and the high-risk population was screened by node gain analysis.Results Four important explanatory variables were screened out by the classification tree model from the 27 candidate variables related to primary renal damage,including retinopathy,fibrinogen,waist-hip ratio (WHR),red blood cell distribution width (RDW).Retinopathy was an major factor of DKD.The probability of macro-albuminuria in retinopathy and WHR ＞ 0.82 group was 43.8％,and if at the same time RDW ＞ 0.14,the probability of macro-albuminuria was 88.9％.Conclusions The classification tree model can analyze factors of the separate stages of proteinuria in type 1 diabetic patients effectively.Retinopathy is the major influential factors of type 1 diabetic patients with proteinuria.
ABSTRACT
Objective To analyze the related factors of micro-albuminuria and macroalbuminuria in type 1 diabetes mellitus (DM) in the classification tree model,and to screen the high risk population of diabetic kidney disease.Methods 394 patients with type 1 diabetes were enrolled in the hospital from 2008 to 2015.According to glomerular filtration rates and urine albumin quantification,the patients were divided into type 1 diabetes group (299 cases),micro-albuminuria group (73 cases) and macro-albuminuria group (22 cases).The classification tree model was utilized to analyze related factors of the different stages of proteinuria,and the high-risk population was screened by node gain analysis.Results Four important explanatory variables were screened out by the classification tree model from the 27 candidate variables related to primary renal damage,including retinopathy,fibrinogen,waist-hip ratio (WHR),red blood cell distribution width (RDW).Retinopathy was an major factor of DKD.The probability of macro-albuminuria in retinopathy and WHR ＞ 0.82 group was 43.8％,and if at the same time RDW ＞ 0.14,the probability of macro-albuminuria was 88.9％.Conclusions The classification tree model can analyze factors of the separate stages of proteinuria in type 1 diabetic patients effectively.Retinopathy is the major influential factors of type 1 diabetic patients with proteinuria.
ABSTRACT
Objective To analyze the impact factors for early renal damage in type 2 diabetic patients by the classification tree model.Methods A total of 601 patients with type 2 diabetes were enrolled.According to glomerular filtration rates and urine albumin quantification,the patients were divided into type 2 diabetes group (418 cases) and early diabetic renal damage group (183 cases).The clinical data of the patients were recorded to analyze the main influential factors for the microalbuminuria of type 2 diabetic patients using the Exhaustive CHAID classification tree algorithm.Results Six important explanatory variables were screened out by the classification tree model from the 34 candidate variables related to early renal damage,including fibrinogen,history of hypertension,retinopathy,Cys C levels,SBP and peripheral neuropathy.Elevated fibrinogen was the main factor.Conclusion The classification tree model can analyze the major influential factors of early renal damage in type 2 diabetic patients effectively,and it can help develop the prevention and treatment methods.
ABSTRACT
Objective To analyze the prevalence,risk factors of kidney disease in type 2 diabetic patients with KDIGO classification of chronic kidney disease,also to study cardiovascular and cerebrovascular diseases and death in these patients,so as to investigate the significance of the KDIGO classification system.Methods One thousand six hundred and forty-five type 2 diabetic patients who were in hospitalization from June 2008 to December 2012 were grouped according to the KDIGO classification of chronic kidney disease and the incidence of vascular disease was analyzed based on the classification.Clinical features were compared between patients with or without kidney disease.The risk factors of kidney disease and the death of diabetic patients were also investigated.Results There were 915 male and 730 female,aged a median (57.86±12.54) years with (6.35±6.30) years duration of diabetes mellitus among the 1645 cases,and 37.2％ of patients had concomitant kidney disease.According to the classi fi cation of CKD,patients in CKD group 3a,group 3b and CKD group 4-5 accounted for 5.7％,3.5％ and 7.6％,while 33.4％ of patients had proteinuria,among which 19.5％ with microalbuminuria,13.5％ with macroalbuminuria.On complications,patients with hypertension accounted for 49.5％,hyperlipidemia 67.7％,diabetic retinopathy 27.4％,cardiovascular and cerebrovascular diseases 18.5％ (coronary artery disease 16.5％,cerebrovascular diseases 8.8％).Statistical difference was detected in the incidence of diabetic retinopathy,coronary artery disease and cerebrovascular diseases between CKD group 3a and 3b (P ＜ 0.05).The duration of diabetes,concomitant hypertention especially with elevated systolic blood pressure,diabetic retinopathy and hyperuricemia were the independent risk factors for type 2 diabetic patients with kidney disease.Age,Scr,complicating cardiovascular and cerebrovascular diseases and advanced CKD stage were the independent risk factors for the death of type 2 diabetic patients with kidney disease.Conclusion KDIGO classification of chronic kidney disease enables better staging of kidney diseases in diabetic patients for management and prognosis.Diabetic patients have a higher prevalence of renal diseases and cardiovascular and cerebrovascular events than the general population.Early control of factors such as blood pressure and serum uric acid can delay the progression of kidney disease,and the predictive role of diabetic retinopathy should be emphasized.
ABSTRACT
The aim of this study is to investigate the growth and proliferation of bone marrow mesenchymal stem cells (BMSCs) three-dimensionally cultured in Pluronic F-127 gel, in order to explore the cellular compatibility of gel and to investigate the feasibility of BMSCs differentiating into adipocytes in gel. Rat BMSCs were isolated from adult bone marrow, and then cultured and amplified in vitro. The BMSCs derived from the 4th passage were seeded on the scaffolds and incubated in adipogenic stimuli culture to differentiate into adipocytes. BMSCs were dispersed into gel and cultured in vitro for two weeks then the status of adhesion, growth and proliferation of the cells were observed. The edipogenic differentiation of the BMSCs was assessed by cellular morphology and further confirmed by Oil Red O staining. BMSCs were able to attach, grow and proliferate well in Pluronic F-127 gel. The BMSCs differentiated into adipocytes in gel in the presence of adipogenic stimuli over a period of 2 weeks. After only 4 days of adipogenic induction, small lipid droplets were observed within BMSCs in gel wells treated with differentiation media. At the end of 14 days, in the presence of differentiation media in gel, the size of the lipid droplets increased to occupy most of the cytoplasm, consistent with differentiation of BMSCs into adipocytes. Lipid droplets in differentiating BMSCs were positively stained with Oil Red O in the presence of differentiation media in the Pluronic F-127 treatment. We demostrated BMSCs incubated in the 3D Pluronic F-127 gel scaffolds could be induced and differentiated into adipocytes. The system for inducing differentiation of BMSCs into adipocytes is promising to apply in the construction of tissue engineering adipose tissue and the repair of fat injury, and Pluronic F-127 gel may be a suitable scaffold for cellular therapy of BMSCs.