Clinicopathological analysis for IgA nephropathy with isolated hematuria and/or mild proteinuria / 中南大学学报(医学版)

To investigate the correlation of different types of urinary abnormalities or different proteinuria and hematuria with the pathological injury of kidney in IgA nephropathy with isolated hematuria and/or mild proteinuria.
 Methods: Patients with primary IgA nephropathy, isolated hematuria and/or mild proteinuria were enrolled in the Department of Nephrology, the Second Xiangya Hospital, Central South University from January 2013 to January 2018. According to the difference of red blood cell count in urinary sediment and quantitative of 24-hour urinary protein (24 h-UP) during renal biopsy, the patients were grouped in 3 ways: a simple hematuria group, a hematuria and proteinuria group, and a simple proteinuria group; a proteinuria I group, a proteinuria II group, and a proteinuria III group; a hematuria I group, a hematuria II group, and a hematuria III group. The clinical parameters such as age, mean arterial pressure, blood urea nitrogen, serum creatinine, blood uric acid, 24 h-UP, and renal pathological damage were compared.
 Results: A total of 157 patients met the inclusion criteria, including 71 males and 86 females. The most common pathological type was focal and/or segmental glomerulosclerosis. The Lee's classification were dominated by grade III and IV, and the renal pathological injury was heavy. Immunoglobulin deposition was dominated by simple IgA deposition. The most common fluorescence intensity of IgA deposition was +++. 97 (61.78%) patients were accompanied by complement deposition and were mainly composed of simple complement C3 deposition. There were 18 patients (11.47%) in the simple hematuria group, 111 patients (70.70%) in the hematuria and proteinuria group, and 28 patients (17.83%) in the simple proteinuria group. Compared with the simple hematuria group, the proportion of patients with mild injury was lower in the simple proteinuria group, and the proportion of patients with moderate-to-severe injuries was increased (χ2=7.053, P=0.008). Compared with the hematuria and proteinuria group, the proportion of patients with mild injury was lower in the simple proteinuria group, and the proportion of patients with moderate-to-severe injury was increased (χ2=4.294, P=0.038). Compared with the proteinuria I group, the proportion of patients with mild injury was lower in the proteinuria III group, and the proportion of patients with moderate-to-severe injury was increased (χ2=5.433, P=0.020). There was no significant difference in the proportion of patients with renal pathological injury among different hematuria groups (P>0.05).
 Conclusion: The clinical manifestations of patients with IgA nephropathy with hematuria and/or mild proteinuria are inconsistent with renal pathological damage. Some patients with mild clinical manifestations have severe renal pathological damage and the renal pathological damage is more serious in simple proteinuria. The more proteinuria, the heavier the renal pathological damage.

Investigation and influencing factors of symptomatic burden in patients with maintenance hemodialysis (MHD) / 中国医师杂志

Objective To investigat the symptom burden of patients with maintenance hemodialysis (MHD) and explore the influencing factors of them,in order to provide the basis for improving the quality of life of these patients.Methods A total of 230 patients with MHD were enrolled in Xiangtan Central Hospital from January 2016 to December 2016,the general condition and disease-related data of them were collected,and the symptom burden of them over the past week was assessed by improved symptom burden scale.Results ① The average age of these patients were (56.8 ± 14.1)years,the sex ratio for men and women was 1.37∶ 1,the median time of dialysis was 24 months,and the frequency of dialysis was (9.16 ± 2.36)times;② The symptom score in MHD patients was (65.72 ± 28.46)points,the top 5 symptoms which had higher incidence were fatigue (72.2％),dry mouth (63.5％),itching (61.3％),falling asleep difficultly (54.3％),drying (49.2％),the top 5 symptoms which were more troubling were restless legs syndrome (2.54 ± 0.73),falling asleep difficultly (2.48 ± 0.83),bone/joint pain (2.45 ± 0.69),fatigue (2.31 ±0.77),easy to wake up (2.16 ±0.78);③ Age,sex,occupation,dialysis age,inorganicphosphorus and serum calcium had an effect on the symptom burden of MHD patients (P ＜ 0.05).Condusions We should focus on patients who are older,female,retired or no occupation,longer dialysis age,calcium or phosphorus metabolism disorders,provide targeted care and treatment,in order to reduce the symptom burden and improve the life quality of them.

Role of M-type phospholipase A2 receptor and its antibody in hepatitis B virus-associated membranous nephropathy / 中南大学学报(医学版)

To examine levels of M-type phospholipase A2 receptor (PLA2R) and its antibody in the patients with hepatitis B virus-associated membranous nephropathy (HBV-MN), and to explore the correlation of PLA2R with laboratory parameters and pathological characteristics.
 Methods: A total of 49 adult patients with biopsy-proved HBV-MN were enrolled in this study. Levels of anti-PLA2R antibody in serum and PLA2R in renal tissue were detected. Patients were assigned into two groups: a positive PLA2R group and a negative PLA2R group. Differences in laboratory parameters and pathological characteristics were compared between the two groups.
 Results: Of 49 patients with HBV-MN, 17 had positive PLA2R expression in renal tissues. In the positive PLA2R group, 10 patients were positive for serum anti-PLA2R antibody. Patients with positive PLA2R expression in renal tissues showed higher levels of 24 hour urinary protein [(4.6±3.9) g/d], serum HbsAg (70.5%) and renal HbsAg expression (71%), while lower level of serum albumin [(24.1±7.5) g/L] than those of the negative group.
 Conclusion: PLA2R is expressed in the renal tissues and serum anti-PLA2R antibody can be detected in some HBV-MN patients. Positive PLA2R expression in renal tissue might be related to HbsAg deposition in serum and renal tissues. Patients with positive PLA2R expression in renal tissue have more severe glomerular sclerosis.

Intervention and therapeutic effect of siRNA-HDAC5 on abnormal histone modification in non-obese diabetic mice / 中南大学学报(医学版)

OBJECTIVE@#To evaluate therapeutic eff ect of siRNA-HDAC5 on non-obese diabetic (NOD) mice by using small interference RNA (siRNA) technique to knock down the expression of HDAC5 in spleen CD4+ T cells.@*METHODS@#NOD mice, 12-weeks old, were randomly divided into 3 groups and were given normal saline, siRNA-Control or siRNA-HDAC5 through caudal vein injection. The spleens and other samples were collected at the 18th, 24th or 30th week. The blood glucose was tested by blood glucose meter. The urinary albumin and serum levels of IL-1, IL-6, IL-18, and TNF-α were detected by ELISA. The mRNA levels of CD11a, CCR5, and CX3CR1 in spleen CD4+ T cells were measured by quantitative Real-time PCR. The HDAC5 protein level in spleen CD4+ T cell was detected by Western blot.@*RESULTS@#Compared with the control group, the siRNA-HDAC5 group showed a significant decrease in blood glucose, urine albumin excretion rate, serum cytokine and the mRNA levels of CD11a, CCR5, and CX3CR1, consist with the decrease in protein level of HDAC5.@*CONCLUSION@#Inhibition of HDAC5 expression in NOD mice could effectively alleviate the onset and development of kidney damage caused by diabetes.

Role of protein phosphatase 2A in renal interstitial fibrosis / 中南大学学报(医学版)

BACKGROUND@#To explore the role of protein phosphatase 2A (PP2A) in renal interstitial fibrosis by using rat model of unilateral ureteral obstructive (UUO) or cell model of human kidney proximal tubular epithelial (HK)-2 cells treated with transforming growth factor-β1 (TGF-β1).
@*METHODS@#1) A total of 15 Sprague-Dawley rats were randomly divided into a sham group, a UUO group and an okadaic acid (OA) treated group (OA group) (n=5 in each group). The OA 
[30 μg/(kg·d)], diluted with 1.8% alcohol, was given to the rats in the OA group through gastric tube after at 72 h after the surgery, while the equal volume of 1.8% alcohol was given to the rats in the sham group and the UUO group. After sacrificing rats, the blood and kidney were collected to detect the renal function and the expression of PP2Ac, fibronectin (FN), collagen-I (Col-I), E-cadherin (E-cad) and α-smooth muscle actin (α-SMA) by immunohistochemistry, Western blot and RT-PCR, respectively; 2) The likely concentration of OA was determined by Trypan blue dye exclusive assay and methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay. The HK-2 cells were incubated with serum-free Dulbecco's modified eagle medium (DMEM) for 24 h; then they were divided into a control group, a TGF-β1 group (treated with 5 ng/mL TGF-β1 for 24 h) and a TGF-β1+OA group (treated with 5 ng/mL TGF-β1 and 40 nmol/L OA for 24 h). The HK-2 cells were collected and the expression of PP2Ac, FN, Col-I, E-cad and α-SMA were detected by Western blot.
@*RESULTS@#1) Compared with the sham group, the BUN and Scr in the UUO group increased (both P<0.05); compared with the UUO group, the BUN and Scr in the OA group decreased (both P<0.05); the expression of PP2Ac, FN, Col-I and α-SMA was up-regulated while the expression of E-cad was down-regulated in the UUO group compared with those in the sham group (all P<0.05). The expression of PP2Ac, FN, Col-I and α-SMA was down-regulated while the expressions of E-cad was up-regulated in the OA group compared with those in the UUO group (all P<0.05); 2) The likely concentration of OA was 40 nmol/L. Western blot showed that the expression of PP2Ac, FN, Col-I and α-SMA was up-regulated while the expressions of E-cad was down-regulated in the TGF-β1 group compared with those in the control group (all P<0.05); the expression of PP2Ac, FN, Col-I and α-SMA were down-regulated while the expression of E-cad was up-regulated in the TGF-β1+OA group compared with those in the TGF-β1 group (all P<0.05).
@*CONCLUSIONS@#PP2A might be able to promote the renal interstitial fibrosis.
.

Abnormal percentage of memory B cells in tonsils and peripheral blood in clinical progression of IgA nephropathy / 北京大学学报(医学版)

Objective: To observe an abnormal expression of humoral immune response induced by memory B cells in tonsils and peripheral blood of patients with IgA nephropathy ( IgAN) , the variation of memory B cells after tonsillectomy , and to discover the role of tonsillectomy in IgAN .Methods: In the study , 28 patients were diagnosed as IgAN via renal biopsy , and 27 patients suffering from chronic ton-sillitis without nephritis and 10 normal human beings were selected as controls .The expression of memory B cells in the tonsils and peripheral blood was tested by flow cytometry , and the same method was used to test the variation of the expression of memory B cells in peripheral blood of patients with IgAN after tonsil -lectomy.Results:In this study , higher percentages of memory B cells were observed in tonsil and pe-ripheral blood of IgAN patients, which were 5.72%±5.26%, 4.92%±5.10%.After tonsillectomy, the percentage of memory B cells was 1 .10%±0 .65%, lower than that before tonsillectomy ( P <0.05).Meanwhile, in tonsils and peripheral blood , the percentage of memory B cells varied with the variation of the urinary findings of the IgAN patients .Conclusion:The percentage of memory B cell in tonsils and peripheral blood could predict disease progression of IgAN to a certain extent .

Changes of plasminogen activator inhibitor-1 and D-dimer during continuous blood purification and related factors / 中南大学学报(医学版)

OBJECTIVE@#To observe the changes of plasminogen activator inhibitor-1 and D-dimer during continuous blood purification (CBP) and related factors.@*METHODS@#Sixteen patients who were diagnosed with multiple organ dysfunction syndrome (MODS) were randomly divided into 2 groups: 8 patients received standard continuous blood purification with heparin anticoagulation, and the other 8 received CBP without anticoagulation. Ten normal blood samples were collected from healthy volunteers as controls. All patients underwent CBP for 8 h. Blood was taken from those patients at 0, 15, 60, 120 and 480 min during the CBP. Plasma plasminogen activator inhibitor-1, D-dimer and serum TNF-α and IL-1β were measured by ELISA.@*RESULTS@#Plasma levels of PAI-1 and D-dimer were increased significantly compared with those in the control group (P<0.05). Plasma level of PAI-1 was reduced (P<0.05) and D-dimer was increased (P<0.05) after the CBP. The level of plasma PAI-1 in the heparin group was significant reduced compared with the group of CBP without anticoagulation (P<0.05). There was negative correlation between the level of PAI-1 and the dosage of heparin used during a CBP session in the heparin group (r=-0.746, P<0.001).@*CONCLUSION@#The level of PAI-1 and D-dimer is higher in patients with MODS than that in the normal controls. After the CBP treatment, there is significant decrease in PAI-1 and increase in D-dimer in both groups. Heparin used during CBP can reduce PAI-1 which intensifies its function of anticoagulation.

Immune pathogenesis of IgA nephropathy and its drugable targets / 中南大学学报(医学版)

IgA nephropathy (IgAN) is recognized as the most common immune complex related to the cause of glomerulonephritis worldwide. The disease is characterized by the predominant deposition of underglycosylated IgA1 in the mesangial area of glomeruli. Dysregulation of the immune system plays an important role in the pathogenesis of IgAN. Abnormalities restricted to T lymphocytes and/or B lymphocytes activation could be a critical causative factor in the over-production of underglycosylated IgA1.

Efffect of plasminogen activator inhibitor-1 and endothelin-1 on the atherosclerosis in the maintenance hemodialysis patients / 中南大学学报(医学版)

OBJECTIVE@#To investigate the effect of plasminogen activator inhibitor-1 (PAI-1), tissue type plasminogen activator (t-PA), and endothelin-1 (ET-1) on the atherosclerosis progress in the maintenance hemodialysis patients.@*METHODS@#We enrolled 19 patients with maintenance hemodialysis (MHD) and 11 healthy people as control. Patients were divided into 2 groups according to their age above or below 40 years old (11 and 8 in each, respectively), whereas the subjects in control group were below 40 years old. All the clinical information of the research subjects was collected: including age, gender, time of hemodialysis, blood pressure, blood urea nitrogen (BUN), and serum creatinine (SCr). Immunohistochemistry and pathological image analysis were used to investigate the pathological changes, calcification and the expression of PAI-1, t-PA, and ET-1 on the blood vessel.@*RESULTS@#Compared with the age-matched healthy control group, there were higher blood vascular media thickness, blood vascular media thickness/diagmeter ratio, blood vascular media thickness area/vascular inter-wall area ratio (P<0.05) and more calcification (P<0.05) in the the internal iliac artery in the chronic renal failure MHD patients. All the results were similar when compared the above 40 years old group with the below 40 years old one in the chronic renal failure MHD patients. There were positive correlation of blood vascular media thickness with age and blood pressure (P<0.05). Expression of PAI-1, ET-1, t-PA on the internal iliac artery vessel was elevated in the chronic renal failure MHD patients compared with the health control (P<0.05). The level of PAI-1 or ET-1 was much higher in the above 40 years old group than the below 40 years old one in the chronic renal failure MHD patients, whereas there was no significant difference in the t-PA expression between the 2 groups (P<0.05). There were positive correlation of PAI-1 or ET-1 expression with age and blood pressure (P<0.05). There were positive correlation of PAI-1 or ET-1 expression with blood vascular media thickness and calcification (P<0.05 or P<0.01). There was no correlation of hemodialysis time with blood vascular media thickness, calcification, PAI-1, t-PA, or ET-1 expressions.@*CONCLUSION@#MHD patients accompany with atherosclerosis which is severer in the patients above 40 years old than the patients below 40 years old. The higher of the blood pressure, the severer of the atherosclerosis. Abnormal expression of PAI-1 plays an important role in the progress of the atherosclerosis in the chronic renal failure MHD patients, whereas t-PA has no function in this process. The level of PAI-1 and ET-1 would be helpful to evaluating the degree of atherosclerosis in the chronic renal failure MHD patients. Hemodialysis time may not be a potential accelerator for atherosclerosis progression.

Meta analysis of infection risks of anti-TNF-αtreatment in rheumatoid arthritis / 中南大学学报(医学版)

Objective:To systematically evaluate the risks of anti-TNF-αtreatment-associated infection, severe infection and tuberculosis in rheumatoid arthritis (RA) patients, and to reduce the infection incidences associated with anti-TNF-αtherapy. Methods:We used Meta analysis to systematically review randomized controlled trials on anti-TNF-αtreatment associated risks of infecion, severe infection and tuberculosis in AR patients.Results:Although no statistically significant differences were detected in TB risk between anit-TNF-αtreatment and the control group (0.5%vs 0.07%;P=0.27, OR=1.85, 95%CI:0.62-5.52), there still existed a clinically obvious elevation of TB risk in monoclonal anti-TNF-αtreatment, which was illustrated by the results that no TB case was reported in the etanercept group, but 11 TBs in 2050 infliximab-treated cases, and 3 TBs in 722 adalimumab-treated cases. The total infection and severe infection risks were also signiifcantly higher in patients receiving anti-TNF-αtreatment (P0.05), while both kinds of monoclonal antibodies of TNF-αblockers showed a signiifcantly elevated infection or severe infection risks (P<0.05). High doses of anti-TNF-αtreatment were associated with statistically increased risks of severe infection (6.0%vs 2.8%, P=0.04, OR=1.68, 95%CI:1.02-2.78). Conclusion:The TB risk of anti-TNF-αtreatment deserves close attention, especially in places with high rate of BCG vaccination and MTb infection. Monoclonal anti-TNF-αtreatment brings higher risks of infection and severe infection than soluble TNF-αreceptor.

RIFLE and AKIN criteria for mortality and risk factors of acute kidney injury in hospitalized patients / 中南大学学报(医学版)

Objective:To evaluate the mortality and risk factors for acute kidney injury (AKI) in hospitalized patients by the risk, injury, failure, loss, end stage kidney disease (RIFLE) and acute kidney injury network (AKIN). Methods:We constructed a retrospective study of all AKI patients in the Second Xiangya Hospital of Central South University between February 2006 and January 2011. The diagnosis and classiifcation of AKI were reconifrmed and categorized by RIFLE and AKIN criteria. To compare the clinical characteristics, mortality and associated risk factors in AKI patients by the RIFLE and AKIN stage, univariate analysis and multivariate logistic regression analysis were performed. Results:The patients were diagnosed as AKI by AKIN (n=1027) or by RIFLE criteria (n=1020). There was no signiifcant difference in the hospital mortality, hospital length stay (days), or the proportion of complete recovery in each stage of AKI patients by RIFLE and AKIN (P>0.05). In the univariate analysis, age, pre-renal causes, proportion of hospital acquired AKI, mechanical ventilation, hypotension, the number of failed organs, acute tubular necrosis-index severity score (ATN-ISS), and the peak of serum potassium ion concentration were signiifcantly higher in the non-survivors than in the survivors (P<0.05). Logistic regression analysis revealed that age older than 65, hospital acquired AKI, hypotension, number of failed organs, ATN-ISS scores, and the peak of serum potassium ion concentration were independent risk factors for hospital mortality. Conclusion:Both RIFLE and AKIN criteria have similar scientiifc value in assessing hospital mortality. AKI stage is associated with the recent prognosis of AKI patients.

Expression of IgA class switching gene in tonsillar mononuclear cells in patients with IgA nephropathy / 中华肾脏病杂志

Objec0tive To investigate the molecular mechanism of the mal-production of IgA and IgA1 by tonsillar mononuclear cells (TMCs) in IgA nephropathy (IgAN) patients by measuring the mRNA expression of Iα-Cα germline transcript and the mRNA and protein expression of activated induced cytidine deaminase (AID) in cultured TMCs stimulated with lipopolysaccharide (LPS) or hemolytic streptococcus (HS) in IgAN patients as well as the chronic tonsilitis patients. Methods Twent-seven IgAN patients admitted into our hospital from Jan.2009 to Feb.2010 were enrolled.Twent-seven patients with chronic tonsillitis without renal disease were selected as control.Tonsillar mononuclear cells were isolated by density gradient centrifugation in lymphocyte separation medium.The amount of IgA or IgA1 secreted in the culture supernatants was determined by specific enzyme-linked immunosorbent assay (ELISA).Expressions of Iα-Cα germline transcript and AID mRNA were examined by real-time PCR.The AID protein was determined by Western blot. Results The production of IgA and IgA1 protein,especially the ratio of IgA1/IgA and the expression of AID protein in TMCs were significantly increased in IgAN group compared with chronic tonsillitis group (all P＜0.05).The IgA and IgA1 level of stimulated TMCs were obviously increased in patients with IgAN compared with control group (P＜0.05).And the expressions of Iα-Cα mRNA,AID mRNA and AID protein were up-regulated significantly in stimulated TMCs (all P＜0.05). Conclusions Both LPS and HS can induce the production of IgA and IgA1 and up-regulate the expressions of AID and Iα-Cα in TMCs of IgAN patients.Our results indicate that the TMCs are capable of producing high level of IgA and IgA1 stimulated by LPS or HS,whuch may be due to the　 incression of AID and Iα-Cα.

Multi-frequency low-dose intravenous iron on oxidative stress in maintenance hemodialysis patients / 中南大学学报(医学版)

OBJECTIVE@#To investigate the effect of different intravenous iron treatment regimens on anemia and oxidative stress in maintenance hemodialysis (MHD) patients.@*METHODS@#A total of 58 MHD patients were randomly divided into a multi-frequency low-dose intravenous iron group (iron sucrose 25 mg, twice a week for 8 weeks, n=19), a less-frequency regular-dose intravenous iron group (iron sucrose 100 mg, once every two weeks for 8 weeks, n=19), and a non-iron group (n=20). Another 20 healthy people served as a control group (n=20). The changes of hemoglobin (Hb), hematocrit (HCT), serum ferritin (SF) and transferrin saturation (TSAT), as well as the oxidative stress parameters of malon-dialdehyde (MDA), superoxide dismutase (SOD) and myeloperoxidase (MPO) were detected before and after the treatment.@*RESULTS@#After 8 weeks, compared with the non-iron group, the levels of Hb, HCT, SF and TSAT in the two iron groups were significantly elevated (P0.05). After the single dialysis, the two iron groups had higher level of serum MDA, MPO and lower level of serum SOD than that of the non-iron supplementation group (P0.05).@*CONCLUSION@#Multi-frequency low-dose intravenous iron can effectively improve anemia in MHD patients, whose acute oxidative stress is lower than that of less-frequency regular-dose intravenous iron, and is a relatively safe and effective intravenous iron treatment regimen.

Clinical significance of saliva urea, creatinine, and uric acid levels in patients with chronic kidney disease / 中南大学学报(医学版)

OBJECTIVE@#To explore the changes and clinical significance of saliva urea, creatinine (Cr), uric acid (UA) in both healthy people and chronic kidney disease (CKD) patients, and to provide a noninvasive, quick, accurate and reliable test to diagnose kindey disease.@*METHODS@#Urea, Cr and UA in the saliva and serum collected from both healthy people and the CKD patients were measured by biochemical analyzer. We calculated the correlation coefficient of Urea, Cr and UA between the saliva and serum, compared the levels of saliva Urea, Cr and UA among CKD patients in different periods, drew the receiver operation characteristic (ROC) curve and analyzed the sensitivity and specificity of saliva Urea, Cr and UA to predict CKD patients in various periods.@*RESULTS@#The concentrations of Urea, Cr and UA in both the saliva and the serum were positively correlated in healthy individuals and CKD patients (r = 0.918, 0.932, 0.840 and 0.984, 0.971, 0.920). The levels of saliva Urea, Cr and UA in the CKD patients were significantly higher than those of healthy people (P<0.05). Saliva Urea, Cr and UA concentrations of middle and late stage CKD patients were obviously higher than those of healthy people and early stage CKD patients (P<0.05). Areas under the curve (AUC) of the ROC of Urea, Cr and UA to diagnose diverse periods of CKD were 0.898, 0.897 and 0.848. The sensitivity was 0.806, 0.776 and 0.704; and the specificity was 0.968, 0.989 and 0.871.@*CONCLUSION@#The levels of Urea, Cr and UA between the saliva and the serum are closely related. The concentration of saliva Urea, Cr and UA can reflect the renal damage, monitor kidney function of the CKD patients, and help diagnose middle to late stage CKD patients. It is a simple, nonivasive and quick method.

New pathologic classification of diabetic nephropathy (retrospective study of 37 cases) / 中南大学学报(医学版)

OBJECTIVE@#To investigate the new pathological classification of diabetic nephropathy (DN) published by Research Committee of the Renal Pathology Society in 2010.@*METHODS@#Renal biopsy specimens were obtained from 37 patients with type 2 diabetes mellitus with micro-albuminuria (MAU) or clinical albuminuria (CAU). These samples were classified according to new pathological classification for DN and new standard scores for interstitial vascular injury.@*RESULTS@#Before the classification, DN was seen in 26 palients. After re-analysis according to the new pathological classification, the patients diagnosed with DN increased to 32. In these 32 DN patients, 1 was classified as type I, 3 as type IIa, 2 as type IIb, 23 as type III and 3 as type IV; 12 patients had mild interstitial injury, 15 had midrange interstitial injury, while 5 had severe interstitial injury.@*CONCLUSION@#The new pathological classification of DN can increase the diagnosis rate and attract more attention to tubular and interstitial damage in DN, contributing to the early diagnosis and treatment of DN.

Effect of norcantharidin on the expression of FN, Col IV and TGF-β1 mRNA and protein in HK-2 cells induced by high glucose / 中南大学学报(医学版)

OBJECTIVE@#To observe the effect of norcantharidin (NCTD) on the expression of mRNA and protein of fibronectin (FN), collagen IV(Col IV) and transforming growth factor-β1(TGF-β1) in human kidney proximal tubular epithelial (HK)-2 cells induced by high glucose.@*METHODS@#HK-2 cells were incubated with serum-free DMEM for 24 h to synchronize cell growth, and then the cells were divided into 4 groups: Group C (5.5 mmol/L D-glucose), Group M (5.5 mmol/L D-glucose + 24.5 mmol/L-mannitol), Group HG (30 mmol/L D-glucose), and Group HG + NCTD (30 mmol/L D-glucose + 0.5-40 mg/L NCTD). Cytotoxicity of HK-2 cells induced by high glucose of NCTD was detected by Trypan blue dye exclusive assay. The effect of NCTD on the proliferation of HK-2 cells in high glucose was determined by MTT. The cells were collected to extract total RNA and protein at 6, 24 and 48 h after the incubation. The expression of FN, Col IV and TGF-β1 mRNA was examined by RT-PCR, and FN, Col IV and TGF-β1 protein was analyzed by Western blot.@*RESULTS@#Trypan blue dye exclusive assay showed NCTD concentrations over 5 mg/L were rather toxic in HK-2 cells. The proliferation of HK-2 cells in high glucose was interrupted by interfered with 5 mg/L NCTD as measured by MTT (P0.05).@*CONCLUSION@#NCTD can downregulate FN, collagen IV and TGF-β1 mRNA and protein expression in HK-2 cells stimulated by 30 mmol/L D-glucose.

Role of SARA in renal tubular epithelial to mesenchymal transition in diabetic nephropathy and its associated mechanism / 中华肾脏病杂志

Objective To determine the effect of smad anchor for receptor activation (SARA) on renal tubular epithelial to mesenchymal transtion (EMT) induced by high glucose and to investigate the associated mechanism.Methods HK-2 cells were exposed to high glucose (30 mmol/L).HK-2 cells were transfected with the plasmids of wild-type SARA [SARA (WT)] or SARA mutant [SARA with SBD deletion,called SARA (dSBD)] and then was stimulated by high glucose.The gene expression was assayed by real-time PCR and the protein expression was detected by Western blotting.Results During the process of high glucose-induced EMT of HK-2 cells,the gene and protein expression of SARA were down-regulated.The expression of TGF-β1 and Smad3 increased after stimulation of high glucose in HK-2.However,the Smad2 mRNA expression increased while its protein expression was down-regulated in a time-dependent manner.Smad2 and Smad3 were activated by high glucose stimulation and Smad3 kept activation for longer time than Smad2.Compared with high glucose group,over-expression of SARA by transfection of SARA (WT) up-regulated the expression of zona occludens(ZO)1 and down-regulated the expression of vimentin (P＜0.05).However,SARA (dSBD) had no such effects on above expressions.The Smad2 protein expression increased along with the over-expression of SARA.Meanwhile,over-expression of SARA prolonged the activation time of Smad2 and shortened the activation time of Smad3.Conclusions TGF-β1 signaling is activated and SARA expression is down-regulated during the process of high glucose-induced EMT in HK-2 cells.Over-expression of SARA can inhibit the EMT via increase of Smad2 protein expression and longer activation time of Smad2.

Expression of serum microRNA-192 in the diabetic nephropathy patients and its impact on TGF-β signaling pathway / 中华肾脏病杂志

Objective To detect the expression of serum microRNA (miR)-192 in diabetic nephropathy (DN) patients and investigate its effects on DN. Methods The serum levels of miR-192 and miR-210 in DN patients,diabetic patients without renal injury and healthy people were detected by real-time quantitative PCR.Down-stream target genes were predicted using TargetScan software and further confirmed by gain of function and loss of function studies in vitro.Mesangial cells were treated with different concentrations of TGF-β1,then miR-192 expressions of cells and supernatant were examined as well. Results Serum miR-192 level significantly decreased in DN patients compared with healthy people (0.41 ±0.09 vs 1.00±0.00,P＜0.01) and diabetic patients without renal injury,while the serum miR-210 levels were not significantly different among three groups.Bioinformatics analysis results showed that some target genes were involved in TGF-β signal pathway.ZBP1,IGF-1 and type Ⅰ collagen (Col Ⅰ ) were chosen and confirmed by Western blotting,which were regulated by miR-192.TGF-β1 decreased the expression of miR-192 in both cells and cell culture supematants. Conclusion Serum miR-192 may promote the progress of DN by regulating TGF-β1 signaling pathway and may be used as a potential biomarker in the diagnosis of DN depending on its certain specificity.

Correlation studies between urinary retinol binding protein and renal tubular damage / 中南大学学报(医学版)

OBJECTIVE@#To study correlation between urinary retinol binding protein (RBP) content and renal tubular damage.@*METHODS@#A total of 1 353 healthy people and 186 patients with renal tubular damage diagnosed by renal biopsy were enrolled. The indicators such as endogenous creatinine clearance rate (Ccr), creatinine(Cr), urinary retinol binding protein(RBP), urinary β(2)-microglobulin(β(2)-MG), urinary N-acety1-beta-D-glucosaminidase (NAG), urine specific gravity(SG), urine osmolality of the 2 groups were examined and compared. Score of tubulointerstitial impairing and all indicators were analyzed by Spearman rank correlation analysis, and the sensitivity and specificity of indicators were calculated.@*RESULTS@#Renal tubular damage was positively correlated with urinary RBP, β2-MG, NAG (r=0.863, P<0.001; r=0.777, P<0.001; r=0.374, P=0.002, respectively), while negatively correlated with urine osmolaling, SG (r=-0.519, P<0.001; r=-0.624, P<0.001, respectively). The specificity and sensitivity for renal tubular damage of RBP were 91.03% and 72.06%.@*CONCLUSION@#RBP is an idea marker for renal tubular damage, and is useful to diagnose renal tubular damage and assess the extent of the damage.

Effect of various concentration calcium on human peritoneal mesothelial cells and protein expression of TNF-α / 中国医师杂志

Objective To study the effect of calcium on human peritoneal mesothelial cells (HPMCs). Methods Proliferation abilities of HPMCs were assessed by tetrazolium salt colorimetry assay (MTT assay) and the levels of LDH in the supernatant were detected in all groups to evaluate the damage of HPMCs. The expression of TNF-α in cytoplasm was detected by immunohistochemistry. Results Calcium enhanced proliferation of HPMCs in a time-dependent manor(P＜0.01), especially calcium with 1.25 mmol/L(0.5098±0.016,0.6763±0.048) and 1.0 mmol/L(0.4853±0.016,0.6678±0.076). Calcium with different concentrations significantly increased the levels of LDH in HPMCs in a time-dependent manor, while the effect of calcium with 1.25 mmol/L was lowest(17.78±1.18,23.60±1.39,P＜0.01). Calcium with 2.0 mmol/L[(42.61±3.29)%] and 1.75 mmol/L[(33.32±1.88)%] significantly up-regulated the expression of TNF-α(P＜0.01) . Conclusions High calcium damaged HPMCs and up-regulated the expression of TNF-αby HPMCs, while physical calcium (1.25 mmol/L)can protect peritoneum and prevent peritoneal fibrosis.