Objective To define more information for familial hematuria by genetic screening in a pedigree with familial hematuria. Methods This was a 4 generation pedigree included 20 family members. The clinical data and laboratory manifestations of the family members were reviewed and collected from medical records. Meanwhile, the peripheral blood samples of 11 family members of the pedigree were collected, and then DNA samples were extracted by salting out method for genetic analysis. For genetic analysis, firstly, three family members including the proband were selected for whole exome sequencing, and the genetic variations were screened according to the sequence variation interpretation guidelines from the American College of Medical Genetics and Genomics (ACMG) for diagnostic sequence interpretation. Then PCR and Sanger sequencing were used to verify the identified pathogenic variants in all family members in the pedigree. Results In the pedigree, 6 female members had persistent hematuria. Among them, 2 died due to end-stage renal disease, 2 died due to non-renal diseases, and 2 maintained stable renal function. One of the two members with stable renal function was diagnosed as IgA nephropathy by renal biopsy. Moreover, diffuse basement membrane lesions were identified in her renal biopsy sample after the electron microscope examination, which resulted in the suspected diagnosis of Alport syndrome. Genetic testing in this pedigree revealed two novel mutations in COL4A4 gene (NM_000092), c.G446T:p.G149V in exon 7 and c.G1249A:p.G417R in exon 20. Conclusion Two novel mutations of COL4A4 gene (c.G446T:p.G149V in exon 7 and c.G1249A:p.G417R in exon 20) in a hematuria pedigree are related with phenotype of familial hematuria.

Objective To summarize the diagnosis features of the prenatal genetic diagnosis of fetal renal cystic disease and to explore the clinical feasibility and significance of prenatal genetic diagnosis of congenital cystic nephrosis. Methods A total of 25 fetuses with congenital renal cystic disease were examined via invasive prenatal diagnosis in Henan Provincial People's Hospital from June 2017 to September 2019. Amniotic fluid samples were extracted by amniocentesis. Chromosomal microarray analysis (CMA) were performed in 17 cases. In addition to CMA, the other 8 cases were analyzed by G-band karyotype. Whole exome sequencing (WES) was performed in 6 cases which got normal results by CMA and karyotype, and highly suspected as hereditary disease. Results Of the 25 fetuses assessed, 4 cases (16.0%) pathogenic copy number variation (pCNV) were found, including 2 cases of 17q12 deletion, 1 case of 10p15.1p14 deletion and 1 case of 4q21.28q22.1 deletion(including PKD2 gene). There were 8 cases without chromosome abnormality by karyotype analysis. Six clinical WES analysis found NPHS1 gene c.1440+1 G>A and c.925G > T mutations were related to Finnish type congenital nephrotic syndrome in 1 case, PKD1 gene c.6878C>T mutation was related to autosomal dominant polycystic kidney disease (ADPKD) in 1 case, and there was no definitive mutation in 4 cases. Conclusions CMA and next generation sequencing are powerful tools for accurate diagnosis, treatment and genetic counseling of fetal congenital renal cystic diseases. For congenital cystic nephropathy, genetic detection is helpful to clarify the etiology, and provide more exactly informations for prognosis evaluation, treatment and family genetic counseling.

Objective To discuss the etiology, clinical manifestations and renal pathological features of acute interstitial nephritis (AIN) in children. Methods The etiology, clinical manifestations, pathological characteristics, clinical effects and outcome of the children with AIN diagnosed by renal biopsy from January 2010 to December 2019 in Nanjing Jinling Hospital were analyzed retrospectively. The Kaplan-Meier method was used to evaluate the kidney survival rate. Cox regression model was built to analyze the risk factors for developing end-stage renal disease (ESRD) at baseline in AIN children. Results A total of 51 children with AIN were diagnosed by renal biopsy, including 36 males and 15 females. The age was (12.94±2.55) years old (2-17 years old). The clinical manifestations of AIN in children were various and lack of specificity. Only 2 cases (3.92%) had triad. All of the 51 children with AIN showed acute renal injury (AKI), accompanied by increased serum creatinine and decreased estimated glomerular filtration rate. The stage of AKI was mainly stageⅢ(33 cases, 64.71%). Infection was the main cause (38 cases, 74.51%) and drug factor was the second cause (27 cases, 52.94%) in children with AIN. Nonsteroidal antiinflammatory drugs (NSAIDs) were the main inducers of drug-induced AIN (18 cases, 35.29%). The interstitial inflammatory cell infiltration or interstitial edema was found in 51 children. The infiltration of inflammatory cells was mainly mononuclear cells (46 cases, 90.20%). After 4 weeks of treatment, 32 cases (62.75%), 11 cases (21.57%) and 8 cases (15.69%) showed complete, partial and no recovery of renal function, respectively. After 12 weeks of treatment, 49 cases (96.08%), 0 cases (0) and 2 cases (3.92%) showed complete, partial and no recovery of renal function, respectively. After an average follow-up of 4.0(2.0-15.0) months, 2 case (3.92%) patients developed ESRD. The cumulative survival rates of ESRD at 1 year and 2 years after renal biopsy both were 100%, and renal survival rates at 5 years and 10 years were 96.55% and 72.41%, respectively. Multivariate Cox regression analysis results showed that N-acetyl-β-D-glucosidase (NAG) enzyme level>17.6 U/g·cr (HR=15.729, 95%CI 1.045-15.977, P=0.042) and IgM deposition in renal tissue (HR=7.523, 95%CI 1.142-9.541, P=0.033) were independent risk factors for developing ESRD in AIN children. Conclusions AKI is the main clinical manifestation of AIN in children. The characteritic of renal pathology in AIN is tubulointerstitial lesions. After active treatment, most of the patients have a good prognosis. Prevention of infection and rational use of drugs are the key to reduce the incidence rate of AIN in children. The N-acetyl-β-D-glucosidase enzyme level>17.6 U/g·cr and IgM deposition in renal tissue are independent risk factors for developing ESRD in AIN children.

Objective To report two cases of post-transplantation lymphoproliferative disorders (PTLD) after kidney transplantation in children and review the literature, and to improve clinicians' understanding of PTLD in children. Methods The clinical data of two children with PTLD admitted to the Children's Hospital of Fudan University were collected and analyzed. The PTLD-related literature of PubMed, Embase, Web of Science, Scopus, Cochrane Library, Wanfang, CNKI, Weipu Database and China Biomedical Literature Service System from the establishment of the database to January 2020 were collected for literature review. Multivariate logistic regression analysis method was used to analyze the influencing factors of prognostic in children with PTLD. Results Both of the patients had negative Epstein-Barr virus (EBV) and Cytomegalovirus (CMV) before transplantation and anti-thymocyte immunoglobulin (ATG) were induced during transplantation. PTLD in case 1 and case 2 was diagnosed at 3 and 12 months after transplantation, respectively, with positive EBV and CMV serological reaction. The pathological diagnosis was monomorphic PTLD in case 1 and the case 2 was clinically considered as non-hodgkin lymphoma. They all received thrapies of immunosuppressive reduction combined with anti-CD20 monoclonal antibody and chemotherapy. PTLD was relieved and graft function was normal in 2 cases, while case 1 died two and half years after transplantation due to intracranial fungal infection. According to the analysis of 56 children (including 2 cases in this study) with PTLD from the literature review, the median time of PTLD from transplantation was 41.8 months. The initial involved organs were digestive tract [17 cases (30.4%)], respiratory system [8 cases (14.3%)], nervous system [7 cases (12.5%)] and pharyngeal lymph ring [7 cases (12.5%)], respectively. The main pathologic type of PTLD was monomorphic [34 cases (60.8%)]. Fifty-six cases were all positive in EBV serological reaction when PTLD was diagnosed. The treatment included immunosuppressive reduction combined with anti-CD20 monoclonal antibody and chemotherapy. Forty-eight cases of PTLD were relieved, while 8 cases lost graft function. Eleven cases died, including 3 cases due to infection and the other 8 cases due to PTLD. Multivariate logistic regression showed that monomorphic PTLD was a risk factor of death for PTLD children (OR=21.616, 95%CI 1.007-464.107, P=0.049). Conclusions PTLD in children with kidney transplantation is mostly associated with EBV infection, and the clinical manifestations are diverse. Monomorphic PTLD has a poor prognosis and high mortality.

Objective To explore the related risk factors of aortic arch calcification (AoAC) and the relationship between AoAC and long-term outcome in maintenance hemodialysis patients. Methods The patients who underwent hemodialysis in the Blood Purification Center of Beijing Shijitan Hospital Affiliated to Capital Medical University from March to June 2015 were recruited. Calcification of the aortic arch was estimated with plain chest radiology. The patients were divided into AoAC group and no-AoAC group. Multivariate binary logistic regression was used to analyze the influencing factors of AoAC. Kaplan-Meier analysis and Cox regression model were used to examine the association between AoAC and adverse prognostic events (all-cause death and cardiovascular events). Results There were 157 hemodialysis patients included in this study, with age of (62.63±15.05) years (30-90 years old) and 85 males (54.14%). The median follow-up time was 54(20, 54) months. There were 99 cases (63.06%) in AoAC group and 58 cases (36.94%) in no-AoAC group. The age, proportion of diabetes history, serum corrected calcium and triglyceride levels in AoAC group were higher than those in no-AoAC group (all P<0.05), while the proportion of using active vitamin D, serum albumin and intact parathyroid hormone level were lower than those in no-AoAC group (all P<0.05). Multivariate logistic regression analysis showed that older age (OR=1.109, 95%CI 1.067-1.152, P<0.001), diabetes (OR=4.110, 95%CI 1.551-10.890, P=0.004), longer dialysis duration (OR=1.026, 95%CI 1.010-1.043, P=0.001), higher systolic pressure (OR=1.039, 95%CI 1.012-1.067, P=0.005) and higher triglycerides levels (OR=1.932, 95%CI 1.148-3.125, P=0.013) were the independent risk factors of AoAC, and higher hemoglobin was a protective factor (OR=0.967, 95%CI 0.938-0.998, P=0.035) of AoAC. Sixty-three cases (63.64%) died, and 78 cases (78.79%) had cardiovascular events in AoAC group. Fourteen cases (24.14%) died, and 12 cases (20.69%) had cardiovascular events in no-AoAC group. Kaplan-Meier analysis showed higher incidence rate of all-cause death (Log-rank χ²=22.499, P<0.001) and cardiovascular events (Log-rank χ²=50.797, P<0.001) in patients with AoAC. Multivariate Cox regression analysis showed AoAC was the independent risk factor of all-cause death (HR=2.003, 95%CI 1.039-3.859, P=0.038) and cardiovascular events (HR=5.642, 95%CI 3.003-10.600, P<0.001). Conclusions Older age, diabetes mellitus, longer dialysis duration, hypertension, higher triglyceride levels and lower hemoglobin are significantly associated with AoAC. AoAC is the independent risk factor of all-cause death and cardiovascular events in maintenance hemodialysis patients.

Objective To investigate the role and mechanism of Nod-like receptor protein 3 (NLRP3) in chronic kidney disease (CKD)-related neointimal hyperplasia (NH) of vessels. Methods Wild type C57BL/6J male mice were randomly divided into normal control group (n=6) and experimental group (n=18), by removal of 5/6 kidney and ligation of left common carotid artery to establish a NH model. After established successfully, the mice in NH experimental group were randomly divided into NH model group, NLRP3 inhibitor group, and drug control group (n=6/group). C57BL/6J male mice with NLRP3 gene knockout group did not do any treatment after the establishment of NH model. After 3 weeks of feeding, the blood and vascular tissue samples of mice were collected. The pathological changes of vascular tissue samples in mice were observed by hematoxylin-eosin staining. The expressions and localization of NLRP3-related protein were observed by immunofluorescence staining. The expression of NLRP3 mRNA in vascular tissue was detected by quantitative real-time PCR. The activity of caspase-1 in vascular tissue was measured by colorimetric method. Human aortic smooth muscle cells (HASMCs) were treated with 10% uremic serum to simulate the body's internal environment during the uremic phase. NLRP3 small interfering RNA (siRNA) was transfected or NLRP3 inhibitor glibenclamide was added to the cell cultures. The expression of NLRP3 mRNA in HASMCs was detected by quantitative real-time PCR. The activity of caspase-1 in HASMCs was detected by colorimetric method. Results Compared with the control group, the levels of serum creatinine and blood urea nitrogen were significantly increased in the NH model group (both P<0.01). The vascular histopathology showed that vascular intima thickened, vascular smooth muscle cells proliferated and hypertrophied, nuclei were deeply stained, and cells arranged disorderly and migrated to vascular intima in the experimental group. Quantitative analysis showed that the ratio of neointima to lumen increased significantly in the NH model group than that in control group (P<0.01). Compared with the control group, the immunofluorescence staining of vascular tissue showed that the expressions of NLRP3, caspase-1, IL-18, IL-1β and proliferating cell nuclear antigen (PCNA) protein in the NH model group increased (all P<0.01), while the expression of α-SMA decreased (P<0.01). NLRP3 was mainly located in vascular smooth muscle cells (VSMCs). VSMCs showed a synthetic phenotype. Compared with the NH model group, the expression of NLRP3, caspase-1, IL-18, IL-1β and PCNA protein in the NLRP3 inhibitor group and NLRP3 gene knockout group decreased (all P<0.01), the expression of α-SMA increased (P<0.01), and the pathological changes of blood vessels alleviated. Compared with healthy serum group, the expression of NLRP3, IL-18, and IL-1β and bromodeoxyuridine (BrdU) uptake in uremic serum-stimulated group were increased (all P<0.01). After transfection of NLRP3 siRNA and addition of glibenclamide, the expression of NLRP3, IL-18, and IL-1β in VSMCs in uremic serum-stimulated group decreased, and BrdU intake decreased (all P<0.01). Conclusions NLRP3 inflammatory bodies play an important role in promoting CKD-related neointimal hyperplasia of vessels, and glibenclamide can effectively reduce neointimal hyperplasia.

Objective To analyze the differentally expressed long non-coding RNA (lncRNA) among mice of different ages and explore the mechanism of kidney aging. Methods Male C57BL/6 mice aged 3-month-old (n=5), 12-month-old (n=5) and 24-month-old (n=5) (each weighting about 25 g) were randomly selected. PAS staining, Masson staining and senescence associated β-galactosidase (SA-β-gal) staining were used to detect the pathology and cell senescence of mice kidney. High throughput sequencing was performed to detect the differentially expressed lncRNA and their fragments per kilobase million. Real-time quantitative PCR was used to verify the differentially expressed lncRNA. Competitive endogenous RNA (ceRNA) network, which consisted of lncRNA, miRNA and mRNA was built. GO and KEGG enrichment analysis method were used to predict the biological function of differentially expressed lncRNA. Results PAS staining and Masson staining showed the development of kidney fibrosis, and SA-β-gal staining positive region was increased significantly as age increased. There were 938 known lncRNA and 542 novel lncRNA differentially expressed among different ages' mouse kidney. Compared with 3-month-old mice, 33 lncRNA were up-regulated and 43 lncRNA were down-regulated in 12-month-old mice. Compared with 3-month-old mice, 130 lncRNA were up-regulated and 91 lncRNA were down-regulated in 24-month-old mice. Compared with 12-month-old mice, 36 lncRNA were up-regulated and 22 lncRNA were down-regulated in 24-month-old mice. The results of qRT-PCR about verified 10 lncRNAs with larger differential expression multiples and higer expression levels were consistent with the sequencing data. GO enrichment analysis showed that the target genes of lncRNA differentially expressed in the three groups were mostly located in the nucleus and cytoplasm, and might play a role by binding to proteins or participate in various protein phosphorylation, cell cycle, transcription, transcription regulation and other processes. KEGG enrichment analysis showed that the target genes of lncRNA differentially expressed in the three group were significantly enriched in Rap1 signaling pathway, FOXO signaling pathway and MAPK signaling pathway, which were closely related to kidney aging. Conclusion There are significant differences in expression of lncRNA among the kidney of different ages mice, which are involved in the occurrence of renal senescence.