Progress of research on the role of CLCNKB gene in classical Bartter syndrome / 中华医学遗传学杂志

Bartter syndrome is an inherited metabolic disorder characterized by hypokalemic alkalosis and high rennin-angiotensin-aldosteronism which can occur at all ages but mainly in childhood. Classical Bartter syndrome is caused by loss-of-function variants in the gene encoding basolateral chloride channel ClC-Kb (CLCNKB), which is a common type of Bartter syndrome characterized with diverse clinical manifestations ranging from severe to very mild. This article reviews the function and mechanism of CLCNKB variants in Chinese population and the genotype-phenotype correlation of CLCNKB variants in classical Bartter syndrome.

Analysis of clinical diagnosis and treatment of 20 children with hypophosphatemic tickets / 中华实用儿科临床杂志

Objective To analyze the clinical diagnosis and treatment data of 20 children with hypophosphatemic rickets (HR) in order to improve the clinical diagnosis and treatment of HR.Methods The retrospective analysis of clinical data of 20 cases with HR who were hospitalized at Children's Hospital of Nanjing Medical University from May,2010 to April,2016 was performed to summarize the clinical characteristics.All patients were analyzed for the phosphate regulating gene with homologies to endopeptidase on the X chromosome(PHEX) gene by direct sequencing.If no mutations were detected,multiplex ligation-dependent probe amplification analysis was performed.Results All of the 20 cases with HR showed different degrees of growth retardation and typical X-ray rickets.After treatment,the clinical features were improved.Height standard deviation score (HSDS) was improved significantly with longer treatment time,and the difference was statistically significant(P =0.027).There was a correlation between the blood phosphorus fluctuation and secondary hyperparathyroidism(P ＜ 0.05).Nineteen cases had PHEX gene mutations.Truncating mutations was the most frequent mutation type,and 4 new mutations were found.Conclusions Clinical characteristics,laboratory test results and X-ray examination are important clinical index for the diagnosis of HR,and PHEX gene test can be used as an important auxiliary diagnostic tool.Early diagnosis and treatment can significantly improve the clinical manifestations of the patients.

Clinical characters and risk factors for Henoch - Schonlein Purpura combined with cardiac damage in children / 中华实用儿科临床杂志

Objective To summarize the clinical characteristics and laboratory test results of children with Henoch - Schonlein purpura(HSP),and further to analyze the risk factors for HSP combined with cardiac damage. Methods The clinical and laboratory tests findings from 707 children diagnosed as HSP at Nanjing Children's Hospi-tal were retrospectively analyzed,who were recruited from November 2011 to December 2012. The possible risk factors for HSP with cardiac damage in children were recorded,including gender,age,predisposing causes,gastrointestinal symptoms,joint pain,kidney disorders,serum electrolytes,anti - streptolysin 〝O〝 test,erythrocyte sedimentation rate, and complement level were summarized. Chi - square test and Logistic regression were performed to analyze the risk fac-tors of cardiac damage in children with HSP. Results Among 707 cases,192(27. 2% )patients were combined with car-diac damage,115 male and 77 female,and the proportion of men to women was 1. 00: 0. 67;age ranged from 11 months to 15 years and 4 months(6 years and 5 months for median age),6 patients ﹤ 3 years old occupying 3. 1% ,103 patients≥3 - 7 years old occupying 53. 7% ,82 patients≥7 - 14 years old occupying 42. 7% ,1 patient≥14 years old occupying 0. 5% ,and the age of onset in preschool and school age. Electrocardiogram(ECG)abnormalities were found in 190 patients,the main manifestations including long Q - T interval,ST - T segment falling down and sinus bradycar-dia,and one or more items of abnormal myocardial enzymes existed in 24 cases;echocardiography was performed in 35 cases of children,but no abnormality was detected,no obvious symptoms such as flustered or chest tightness or precor-dial distress. Statistical analysis showed that gender,predisposing causes,mixed HSP,complement level were related to the incidence of cardiac damage in children with HSP(P ﹤ 0. 05). Furthermore binary Logistic regression identified that in male patients,the ratio of X1 vs OR ratio was 0. 654(95% CI 0. 462 - 0. 926,P ﹤ 0. 05),for predisposing causes,the ratio of X2 vs OR ratio was 2. 63(95% CI 1. 838 - 3. 765,P ﹤ 0. 001),for mixed HSP,the ratio of X3 vs OR ratio was 2. 452(95% CI 1. 301 - 4. 621,P ﹤ 0. 01),which were independent factors for cardiac damage in chil-dren with HSP. Conclusions ECG and/ or myocardial enzyme spectrum abnormalities are the main clinical ma-nifestations of cardiac damage in children with HSP. Male patients,predisposing causes of the respiratory tract infec-tion,mixed HSP and hypocomplementemia were high risk factors in the development of cardiac damage,which require special consideration clinically,and earlier ECG and myocardial enzymes examination,early diagnosis and treatment are necessary to avoid the occurrence of severe cases.

Clinical and immunological features of 45 children with lupus nephritis / 临床儿科杂志

Objective To analyze the clinical and immunological features of 45 pediatric patients with lupus nephritis (LN). Methods Forty-five LN patients were included in this study. Clinical, pathological data and immunological parameters were retrospectively analyzed. Results Forty-five LN patients had 6 males and 39 females, with the mean onset age of (10.9 ± 2.8) years. Acute nephritis was the most common type, accounting for 42.2%. Nephrotic syndrome accounted for 31.1%. Renal biopsy showed class II (17.8%), III (4.4%), IV (48.9%), V (2.2%), V+III (6.7%)and V+IV (13.3%)in 42 cases. The remis-sion rate reached 91.1%in the early therapeutic stage, and 15.0%patients recurred after 24-month follow-up. Conclusions The clinical manifestations of LN children are diverse. The renal pathology is complex. The clinical manifestations in part of the chil-dren are not consistent with renal pathology.

Detection of urinary α1-antitrypsin for predicting response to glucocorticoid therapy in children with primary nephrotic syndrome / 中华肾脏病杂志

Objective To detect the α1-antitrypsin (AAT) concentration in urine samples of children with primary nephrotic syndrome (PNS) before initiation of glucocorticoid treatment,in order to verify whether it could predict the response to glucocorticoid-based therapy.Methods Forty-three children diagnosed as PNS initially were chosen as subjects,namely steroid-sensitive nephrotic syndrome (SSNS) and steroid-resistant nephrotic syndrome (SRNS) depending on reaction to glucocorticoid therapy four weeks later,and 15 healthy children serving as normal control.The mid stream of the first morning urine samples were collected from children before taking glucocorticoid.ELISA kit was used to quantify the urinary AAT concentration which was revised by urine creatinine further.The data of urine AAT/Cr were expressed as median with interquartile range.Data analysis was performed using the SPSS 17.0.Results AAT was absent in urine samples of normal healthy children,and there were no statistic differences of the AAT concentrations in urine between children with SSNS and SRNS [(30.4+4.5) mg/L vs (31.8+4.6) mg/L,t=-1.0,P=0.33].The level of urine AAT/Cr in children with SRNS was higher than that in children with SSNS [0.049(0.028-0.073) vs 0.028(0.022-0.036),Z=2.4,P=0.02].Among the laboratory parameters of the two subgroups before taking glucocortiod,the levels of platelet,blood white cell count,serum globulin,urine white cell count,urine red cell count,urine IgG and urine α1-microglobulin were significantly different (P＜0.05).Three parameters that included urine AAT/Cr (OR=6.81 × 1028,P=0.O05),serum globulin (OR=1.69,P=0.01) and urine α1-microglobulin (OR=1.05,P=0.009) further entered the logistic regression model to predict the SRNS independently.The ROC curve based on the level of the urine AAT/Cr was constructed,and the area under the curve (AUC) was 0.72.When the cutoff value of urine AAT/Cr was 0.035,the sensitivity and specificity of the urine AAT/Cr prediction were 68％ and 75％ respectively (Youden' s index 0.43).The AUC that based on the logistic regression model which included urine AAT/Cr,serum globulin and urine α1-mieroglobulin was improved to 0.94,and the sensitivity and specificity of the model prediction were 95％ and 83％ respectively (Youden' s index 0.78).There was no significant difference of the urine AAT/Cr level among the different pathological types of the children undergoing renal biopsy.Conclusions There are no statistic differences of the AAT concentrations in urine between children with SSNS and SRNS.The level of urine AAT/Cr is significantly higher in the SRNS than that in the SSNS which can be as a candidate biomarker to predict the response to glucocorticoid-based therapy.It has a better prediction efficacy based on the model which includes urine AAT/Cr,serum globulin and urine α1-microglobulin.

Oxidative stress-dependent Ras-ERK activation involves in aldosterone-induced mesangial cell proliferation / 中华肾脏病杂志

Objective To investigate the role of oxidative stress-dependent Rasextracellular signal-regulated kinase (ERK1/2) signaling in aldosterone (ALDO)-induced mesangial cell proliferation. Methods The incorporation of 3H-thymidine (3H-TdR) and cell count were used as the measure of mesangial cell (MC) proliferation.Western blotting was used to detect the activation of Ki-RasA,c-Raf,MEK1/2,ERK1/2 and PI3K. Results Aldosterone significantly induced human mesangial cell proliferation,and anti-oxidant N-Acetylcysteine (NAC),catalase,and super oxide dismutase (SOD) significantly inhibited ALDO-induced mesangial cell proliferation (P＜0.01,respectively).Stimulation by ALDO for 3 h,Ki-RasA,c-Raf,MEK1/2,and ERK1/2 activity increased by 4.05-, 3.62-, 4.52-, and 3.40-fold compared with control group (P ＜0.01,respectively).NAC almost completely blocked ALDO-induced Ki-RasA,c-Raf,MEK1/2,and ERK1/2 activation (P＜0.01,respectively).Ki-RasA siRNA dose-dependently inhibited Ki-RasA expression, ALDO-induced Ki-RasA activation, and mesangial cell proliferation (P ＜0.01,respectively).c-Raf inhibitor GW5074 and MEK1/2 inhibitor PD98059 also reduced ALDO-induced mesangial cell proliferation by 65％ respectvely (P＜0.01).Ki-RasA siRNA had no effect on ALDO-induced PI3K phosphorylation.Combining LY294002 and PD98059 completely blocked ALDO-induced mesangial cell proliferation (P＜0.01). Conclusions ALDO-induced Ki-RasA-c-Raf-MEK-ERK signaling activation is dependent on reactive oxygen species (ROS) production,which mediates ALDO-induced mesangial cell proliferation.Inhibition of both ERK1/2 and PI3K signaling simultaneously completely blocks ALDO-induced mesangial cell proliferation.

Clinicopathological features and treatment of C1q nephropathy in children / 中华肾脏病杂志

Objective To investigate the clinicopathological characteristics and treatment of C1q nephropathy in children.Methods Data of 23 C1q nephropathy cases in Nanjing Children's Hospital during recent eight years were retrospectively reviewed. Results The incidence of C1q nephropathy was 4.78％ in primary glomerulonephritis proven by biopsy.Among 23 patients,15 were boys and 8 were girls.The mean age at onset was (5.0±3.4) years old with a range of 0.9-12.4 years.The clinical manifestations included nephrotic syndrome(NS) in 18 cases (78.3％),nephrotic-range proteinuria in 4 cases(17.4％) and microhematuria in 1 case.Two patients with NS and one patient with nephrotic-range proteinuria also presented microhematuria.One patient with NS who received oral herbal medicine for two weeks developed acute renal insufficiency at the same time of diagnosis.Three cases had a family history of kidney disease,among them two patients(presented nephrotic range proteinuria) were siblings,their father had proteinuria as well,and routine genetic examination confirmed familial Denys-Drash syndrome in association with C1q nephropathy.One NS patient's sister had nephrotic-range proteinuria too,but renal biopsy was not performed.No patient had hypertension.None of the patients had low C3 or C4 levels,and serological markers of systemic lupus erythematosus were absent.Light microscopy showedminimalchangedisease (MCD)in13cases (56.5％), mesangialproliferative glomerulonephritis(MsPGN) in 6(26.1％) and focal segmental glomerulosclerosis(FSGS) in 4(17.4％).Immunofluorescence displayed C1q co-deposits of IgG(78.3％),IgM(78.3％),IgA (34.8％) and C3 (47.8％),and a full-house pattern was found in 6 patients (26.1％).Electron microscopy revealed 4 out of 19 had mesangial deposits,except for 4 patients whose glomerulus could not be found.Children with either NS(18 cases) or nephrotic-range proteinuria(2 cases)received prednisone,among them,15 were steroid-resistant,4 were steroid-dependent,only 1 was steroid-sensitive.Those with steroid-resistant(15 cases) or steroid-dependent(3 cases) received further immunosuppression with cyclophosphamide(CTX) or cyclosporine A (CsA).One NS case of steroid-dependent received prednisone re-induction therapy.The siblings associated with DenysDrash syndrome and one case presented microhematuria were commenced on angiotensin-converting enzyme inhibitor(ACEI).Of the 19 patients with sufficient follow-up date,15 cases (78.9％)achieved complete remission,2 cases(10.5％) achieved partial remission,and 2 cases (10.5％) were ineffective.Median follow-up was 15 months.Remission of the NS occurred in 94.4％ (17/18)while nephrotic-range proteinuria was 50.0％(2/4).Remission of MCD was 100.0％,MsPGN was 83.4％(5/6),but FSGS was only 50.0％(2/4).Conclusions C1q nephropathy is rare,and often manifests as steroid-resistant or steroid-dependent NS and nephrotic-range proteinuria.The most common histological feature is MCD,and some as MsPGN or FSGS.A combination of prednisone and immunosuppressive agent is always effective for MCD and MsPGN,but FSGS always has a poor response.

Analysis of clinicopathology and prognosis of childhood Henoch-Schonlein purpura nephritis with diffused endothelial cell proliferation / 中华肾脏病杂志

Objective To investigate the clinicopathological characteristics and prognosis of Henoch-Schonlein purpura nephritis with diffused endothelial cell proliferation (DEP-HSPN) in children. Methods Data of 8 DEP-HSPN cases in Nanjing Children's Hospital within recent ten years were retrospectively reviewed. The clinicopathological features, efficacy and prognosis were compared between DEP-HSPN cases and 48 cases of non-DEP-HSPN. Non-DEP-HSPN cases were divided into two groups according to the clinical classification or the pathological classification.Results (1) In DEP-HSPN, HSP developed nephritis within 4 to 15 days after the initial onset of purpuric rashes. Hematuria was present in all the 8 patients. The main clinical manifestation of DEP-HSPN was nephritic-nephrotic syndrome (4 cases), nephrotic level proteinuria (3 cases) and acute nephritic syndrome (1 case). Four cases had macrohematuria. Six cases had abdominal symptoms and two cases had arthritis. Pathology of all the cases showed grade Ⅲ-b lesion with diffused endocapillary proliferation and segmental necrotizing lesion of the capillary wall, always accompanied with intraglomerular inflammatory cell infiltration. Crescent was found in 4 cases. (2)Compared to non-DEP-HSPN grades Ⅲ, DEP-HSPN showed a shorter course of disease.Macrohematuria, heavy proteinuria, nephritic-nephrotic syndrome, and segmental necrotizing lesion of capillary wall were more common in DEP-HSPN. Compared to non-DEP-HSPN with nephrotic level proteinuria, DEP-HSPN had a lower rate of crescent. (3) Methylprednisolone pulse therapy in early stage, then prednisone combined with cyclophosphamide were used in the treatment of DEP-HSPN.After an average follow-up period of seven months, one patient showed complete remission, five showed persistent microhematuria, and two showed persistent microhematuria accompanied with minor proteinuria. No significant difference of prognosis was found between DEP-HSPN and nonDEP-HSPN. Conclusions DEP-HSPN has an acute onset. The main clinical manifestation of DEP-HSPN is nephritic-nephrotic syndrome and nephrotic level proteinuria, always accompanied with macrohematuria. Immunosuppressant treatment in the early stage of disease is effective for a short-term outcome.

ROS-EGFR-JNK-AP-1 signaling pathway involves in angiotensin Ⅱ-induced human mesangial cells proliferation / 中华肾脏病杂志

Objective To elucidate whether Ang Ⅱ indnces the proliferation of mesangial cells through ROS-EGFR-JNK-AP-1 signaling pathway. Methods The incorporation of 3H-thymidine (3H-TdR) and cell count were used to measure mesangial cell (MC) proliferation. ROS production was determined by DCFDA fluorescence. EGFR and JNK activation was assayed by Western blot. Results Ang Ⅱ significantly enhanced ROS production in mesangial cells, which was up-regulated by 2.26 folds of control group after incubation with Ang Ⅱ for 60 min. Ang Ⅱ induced EGFR phosphorylation in dose- and time-dependent manner, with the peak (3.96 folds increase) at 30 min. EGFR phosphorylation was significantly blocked by AT1R antagonist losartan, antioxidant NAC, and NADPH oxidase inhibitor apocynin and DPI. EGFR antagonist AG1478 significantly inhibited Ang Ⅱ-induced mcsangial cell proliferation. Losartan, NAC, apocynin, DPI, and AG1478 ahnost abolished Ang Ⅱ-induced JNK activation. Conclusions ROS-EGFR-JNK-AP-1 signaling pathway is involved in Ang Ⅱ-induced mesangial cell proliferation. Apocynin and AG 1478 may be used as new therapy.

Effect of curcumin on extracellular matrix accumulation in the glomeruli in nephrotoxic sera nephritis rats / 中西医结合学报

OBJECTIVE: To observe whether curcumin could inhibit the accumulation of the collagen IV and fibronectin in the glomeruli in nephrotoxi sera nephritis rats. METHODS: Seventy-two healthy male Sprague-Dawley rats were divided into three groups, with 24 animals in each group. For normal control group, normal saline (0.5 ml/d) was injected through intra-caudal-vein for two days, and at the same time normal saline (0.5 ml/kg) was also daily administered intraperitoneally. For nephrotoxic sera nephritis group, nephrotoxic sera (0.5 ml/d) was injected through the tail vein for two days and dimethyl sulfoxide (0.5 ml/kg) was given intraperitoneally daily. For curcumin group, nephrotoxic sera was injected as above and meanwhile curcumin (50 mg.kg(-1).d(-1)) was administered intraperitoneally every day. Six rats in each group were killed on the 3rd, 7th, 14th and 28th day. Their renal tissue was fixed in 10% formalin for examining the expression of collagen IV and fibronectin. RESULTS: Minimal staining of collagen IV and fibronectin was detected in the basement membrane of normal control rats glomeruli. In the nephrotoxic sera nephritis rats and curcumin treated nephrotoxic sera nephritis rats, the accumulation of collagen IV and fibronectin was increased progressively, with significant difference in the accumulation of collagen IV (P<0.01) between these two groups at the same time points, while the significant difference in fibronectin accumulation (P<0.05) appeared only after the 7th days. CONCLUSION: Curcumin can reduce the accumulation of collagen IV and fibronectin in the glomeruli. Hence we postulated that curcumin might have beneficial effect for retarding glomerulosclerosis.

CD40L expression on the CD4~+T-cells in children with Kawasaki disease and it’s correlation with coronary artery lesion / 中国免疫学杂志

Objective:To explore the pathogenesis of coronary artery lesions in Kawasaki disease(KD) by detecting expression of CD40L on T-cells from patients with KD.Methods:Blood samples were collected from 26 patients with KD before and after intravenous immunoglobulin(IVIG) treatment. Age-matched febrile 16 children with various diseases were studied in parallel as controls. Age-matched normal control 15 children were studied as controls. CD40L expression on T-cells was detected by flow cytometry, soluble E-selection and soluble CD40L levels were measured by enzyme-linked immunosorbent assay.Results:CD40L expression on CD4+T-cells and soluble E-selection levels were significantly higher in patients with KD than that in the febrile control(FC) group and normal group(P