Clinical phenotype and genotype analysis in 9 children with nephronophthisis / 中华肾脏病杂志

Objective:To investigate and analyze the clinical phenotypes and genotypes in children diagnosed with nephronophthisis (NPHP), and to provide references for clinical diagnosis.Methods:Clinical data of 9 children with NPHP diagnosed by genetic testing in the Department of Nephrology, Wuhan Children′s Hospital from April 2017 to January 2022 were retrospectively collected. The clinical characteristics and genetic test results were analyzed.Results:The median onset age was 11.2(3.4, 14.2) years old in 9 patients, including 5 females and 4 males. There were 8 cases of glomerular proteinuria, 8 cases of renal tubular proteinuria, and 7 cases of reduced urinary gravity in 9 patients. All the children had varying degrees of impaired renal function at the time of diagnosis. Seven cases entered chronic kidney disease (CKD) stage 5, 1 case entered CKD stage 3, and 1 case entered CKD stage 4 at the time of diagnosis. All the children had renal ultrasound abnormalities of varying degrees: size change (3/9), echo enhancement (8/9) and cysts (3/9). Extrarenal phenotypes were present in 3 children. Genetic test showed that 6 patients had mutation of NPHP1 gene, 1 patient had mutation of WDR19 gene, 1 patient had mutation of NPHP3 gene and 1 patient had mutation of NPHP5 gene. Conclusions:Deletion mutation of NPHP1 gene is the most common, while NPHP3, NPHP5 and extremely rare WDR19 mutations have also been found in NPHP patients. The clinical manifestations of NPHP are not typical, so it is necessary to find a specific diagnosis method in the early.

Analysis of correlation between clinical phenotypes and genotypes in children with distal renal tubular acidosis / 中华实用儿科临床杂志

Objective:To analyze the correlation between clinical phenotypes and genotypes in 6 children with primary distal renal tubular acidosis (dRTA).Methods:The clinical data of 6 children confirmed as dRTA in Wuhan Children′s Hospital, Tongji Medical College, Huazhong University of Science & Technology from November 2017 to August 2019 were collected, and related auxiliary examination was performed to assess their growth and development.The venous whole blood was reserved for Trio whole exome sequencing, and full spectrum genetic disease accurate diagnosis cloud platform was applied to systematic data screening and analysis.The suspected mutations were checked by Sanger sequencing, and then the role of protein was predicted by software.Results:Clinical manifestations, signs and auxiliary examination results of the 6 children accorded with the diagnostic criteria of dRTA, and the prominent characteristics was growth retardation.One case had knee valgus, one had osteoporosis, and the auxiliary examination results showed that both of them had alkaline urine, metabolic acidosis, and hypokalemia.Three children had nephrocalcinosis, and 2 children had nephrolithiasis.The parents of the 6 patients were all normal without phenotypes.Mutations in the SLC4A1 gene were identified in 4 patients, including 1 child with a reported homozygous autosomal recessive missense mutation(c.2102G>A, p.G701D), who had dRTA and hemolytic anemia, and 3 children with the reported de novo heterozygous autosomal dominant missense mutation(c.1766G>A, p.R589H, c.1765C>T, p.R589C), whose age at diagnosis was related to abnormal renal imaging.Compound heterozygous autosomal recessive mutations in the ATPV1B1 gene were identified in 1 patient, and they were novel heterozygous missense mutations (1153C>A, p.P385T and c. 806C>T, p.P269L). A novel homozygous autosomal recessive missense mutation was identified in 1 patient in the ATPV0A4 gene(c.1899C>A, p.Y633X, 208). Conclusions:Mutations in SLC4A1, ATP6V1B1, ATP6V0A4 genes are identified as the main causes of the primary dRTA, and the phenotypes was related to the mutation features and genotypes.Genetic test should be conducted on patients suspected as dRTA for early molecular diagnosis, thereby improving clinical phenotypic screening and individualized treatment.

Targeting peptide-mediated 131I-polyamidoamine in medullary thyroid carcinoma bearing mice / 中华核医学与分子影像杂志

Objective@#To construct 131I-the fifth generation polyamidoamine (PAMAM(G5.0)) with targeting peptide Ser-Arg-Glu-Ser-Pro-His-Pro (SRESPHP; SR) or Gly-Pro-Leu-Pro-Leu-Arg (GPLPLR; GP) and double targeting peptide SR/GP, and evaluate the targeting ability in medullary thyroid carcinoma (MTC) model.@*Methods@#PAMAM(G5.0), PAMAM(G5.0)-SR, PAMAM(G5.0)-GP and PAMAM(G5.0)-SR/GP were radiolabeled with 131I by chloramine T method. The radiolabeled yield and radiochemical purity were determined by thin layer chromatography. MTC xenografts were developed and the percentage radio-activity of injection dose per gram of tissue (%ID/g) in tumor and organs was measured at 24 h post-injection. Region of interest (ROI) was drawn and the tumor/non-tumor (T/NT) ratios at 4, 8 and 24 h post-injection were calculated and compared among different groups. One-way analysis of variance, repetitive measurement analysis of variance and Dunnett-t test were used to compare the data of different groups. The relationship between %ID/g and T/NT was analyzed with Pearson correlation.@*Results@#The radiolabeled yield was more than 75% and radiochemistry purity was more than 90%. The difference of %ID/g at 24 h post-injection was significant (F=14.400, P<0.001) in tumors of all groups. The radioactive uptake in tumor of 131I-PAMAM(G5.0)-SR group was the highest at 24 h post-injection((1.80±0.18) %ID/g). There were significant differences of T/NT ratios among different groups(F=4.776, P<0.05)and between different time points(F=8.630, P<0.05). Compared with negative control group (Na131I), the T/NT ratios significantly increased in 131I-PAMAM(G5.0)-SR group at 4, 8 and 24 h post-injection (t=4.169, 7.123 and 4.032, all P<0.05) and in 131I-PAMAM(G5.0)-GP group at 4 h post-injection (t=5.893, P<0.05). The T/NT ratio in 131I-PAMAM(G5.0)-SR group was higher than that in 131I-PAMAM(G5.0)-GP group at 24 h post-injection (t=2.871, P<0.05).@*Conclusions@#PAMAM(G5.0)-SR, PAMAM(G5.0)-GP and PAMAM(G5.0)-SR/GP can target the MTC models. 131I-PAMAM(G5.0)-SR has the best biological properties and may provide a new precision method for MTC diagnosis, treatment and prognosis evaluation.

Report of papillorenal syndrome in a family and literature review / 中华肾脏病杂志

Objective To investigate the clinical manifestations and genetic features of children with papillorenal syndrome caused by PAX2 gene mutation.Methods Clinical manifestations,imaging changes and sequencing data were collected and analyzed from a family with papillorenal syndrome who were diagnosed in Wuhan Children's Hospital in February 2018."PAX2","papillorenal syndrome" and "renal coloboma syndrome" were used as key words to search in China National Knowledge Infrastructure,Wangfang Data Knowledge Service Platform,PubMed and Human Gene Mutation Database up to April 2018.Results A ten years old girl was admitted due to "edema and urine output decreased for one week".Lab showed BUN 25.30 mmol/L,Scr 766.5 μmol/L,Urine protein 3.6 g/24 h.Imaging examination showed bilateral vesical and ureter reflux combined with left duplex kidney and duplication of ureter.Developmental dysplasia of the left hip was also found.The father of the patient had been diagnosed with chronic kidney disease for 10 years and on hemodialysis for 6 years.Next generation sequencing revealed that both the father and daughter carried a heterozygous nonsense mutation in the exon3 c.219C ＞ G(p.Y73X) of PAX2.No Chinese literature ever was reported about papillorenal syndrome.Ninety-four articles in English were retrieved and 177 patients with papillorenal syndrome were confirmed by gene analysis with a total of 92 PAX2 variants.Ten nonsense mutations had been reported.Developmental dysplasia of the hip (DDH) never be reported before.Conclusion Papillorenal syndrome caused by PAX2 mutation can mainly manifest as abnormal development of both kidney and optic nerve,which may be accompanied by other systemic abnormalities,it is rarely reported in China.DDH may be a new phenotype of papillorenal syndrome.

Targeting peptide-mediated 131I-polyamidoamine in medullary thyroid carcinoma bearing mice / 中华核医学与分子影像杂志

Objective To construct 131 I-the fifth generation polyamidoamine (PAMAM(G5.0)) with targeting peptide Ser-Arg-Glu-Ser-Pro-His-Pro (SRESPHP;SR) or Gly-Pro-Leu-Pro-Leu-Arg (GPLPLR;GP) and double targeting peptide SR/GP,and evaluate the targeting ability in medullary thyroid carcinoma (MTC) model.Methods PAMAM(GS.0),PAMAM(GS.0)-SR,PAMAM(GS.0)-GP and PAMAM(GS.0)-SR/GP were radiolabeled with 131I by chloramine T method.The radiolabeled yield and radiochemical purity were determined by thin layer chromatography.MTC xenografts were developed and the percentage radio-activity of injection dose per gram of tissue (％ID/g) in tumor and organs was measured at 24 h post-injection.Region of interest (ROI) was drawn and the tumor/non-tumor (T/NT) ratios at 4,8 and 24 h post-injection were calculated and compared among different groups.One-way analysis of variance,repetitive measurement analysis of variance and Dunnett-t test were used to compare the data of different groups.The relationship between ％ID/g and T/NT was analyzed with Pearson correlation.Results The radiolabeled yield was more than 75％ and radiochemistry purity was more than 90％.The difference of ％lD/g at 24 h post-injection was significant (F=14.400,P＜0.001) in tumors of all groups.The radioactive uptake in tumor of 131I-PAMAM (G5.0)-SR group was the highest at 24 h post-injection ((1.80± 0.18) ％ID/g).There were significant differences of T/NT ratios among different groups (F =4.776,P＜ 0.05)and between different time points (F =8.630,P＜0.05).Compared with negative control group (Na131 I),the T/NT ratios significantly increased in 131I-PAMAM(G5.0)-SR group at 4,8 and 24 h post-injection (t=4.169,7.123 and 4.032,all P＜0.05) and in 131I-PAMAM(G5.0)-GP group at 4 h post-injection (t =5.893,P＜0.05).The T/NT ratio in 131I-PAMAM (G5.0)-SR group was higher than that in 131 I-PAMAM (G5.0)-GP group at 24 h post-injection (t=2.871,P＜0.05).Conclusions PAMAM(G5.0)-SR,PAMAM(G5.0)-GP and PAMAM(G5.0)-SR/GP can target the MTC models.131I-PAMAM(G5.0)-SR has the best biological properties and may provide a new precision method for MTC diagnosis,treatment and prognosis evaluation.

Assessment of effects of 7,8-dihydroxyflavone on the striatum in normal cynomolgus monkeys with 99TCm-TRODAT-1 SPECT/CT imaging / 中华核医学与分子影像杂志

Objective To assess the effects of 7,8-dihydroxyflavone (7,8-DHF) on the striatum (ST) in normal cynomolgus monkeys using 99Tcm-TRODAT-1 imaging.Methods A total of six healthy female cynomolgus monkeys were included in this study.Three of them were fed with normal food (control group),and the other three were given oral administration of 7,8-DHF in addition to normal food (experimental group).The SPECT/CT imaging was performed at different time after 99Tcm-TRODAT-1 injection.The ROI of ST was drawn on images of 3 consecutive transverse slices that could be visualized best.The cerebellum (CB) was taken as the background reference area.The radioactivity uptake ratios of ST/CB at 1,3,4 and 5 h were calculated respectively.Paired-t test was used to analyze the data.Results ST radioactive uptake ratios showed continuing increase on the delay images.ST/CB uptake ratios of the control group at 1,3,4 and 5 h were 1.43±0.04,1.82±0.06,2.04±0.12,2.42±0.23,respectively,and those of the experimental group were 1.35±0.08,2.40±0.09,2.74±0.13 and 3.25±0.15 respectively.There was no significant difference between the two groups at 1 h (t =2.57,P＞0.05),while ST/CB uptake ratios of the experimental group at 3,4 and 5 h were significantly higher (t values:2.77,2.87 and 2.92,all P＜0.05).Conclusion 99Tcm-TRODAT-1 SPECT/CT imaging can be used to assess the DAT activation effect by 7,8-DHF on ST of cynomolgus monkeys.

Efficacy and safety of multiple target therapy for lupus nephritis in children / 中华实用儿科临床杂志

Objective To investigate the clinical efficacy of combined use of calcineurin inhibitor in the treatment of lupus nephritis in induction or maintenance,which is resistant to mycophenolate mofetil.Methods Sixty-six cases of children with lupus nephritis were selected from February 2014 to September 2016 in Huazhong University of Science and Technology,Tongji Medical College Affiliated Wuhan Children's Hospital.The randomized method was used to divide them into the control group and the observation group randomly.Among them,31 cases in the control group were given glucocorticoid,cyclophosphamide combined with traditional medicine for treatment;35 cases in observation group were given glucocorticoid,mycophenolate mofetil,tacrolimus (calcine phosphatase inhibitor) multi-target therapy for treatment.The clinical effect of 2 groups before and after treatment were compared,and the incidence of adverse reactions in the treatment of 2 groups of children were compared.Results After treatment,the levels of systemic lupus erythematosus disease activity index (SLEDAI),serum creatinine and 24 h urine protein [(6.05 ± 1.04) scores,(45.08 ± 18.52) μmol/L,(0.96 ±0.30) g/L] in the observation group were lower than those in the control group [(11.09 ±2.33) scores,(95.33 ±36.74) μmol/L,(2.05 ±0.74) g/L],and the differences were statistically significant (t =3.097,3.356,3.871,all P ＜ 0.05).Serum complement C3,plasma albumin levels [(1.05 ± 0.28) g/L,(63.24 ± 12.98) g/L] were higher than those in the control group [(0.34 ±0.10) g/L,(35.45 ±6.74) g/L],and the differences were statistically significant (t =4.124,3.567,all P ＜ 0.05).After treatment,the levels of serum complement C3 and plasma albumin were significantly higher between 2 groups than those before treatment,the differences were statistically significant (all P ＜ 0.05).The incidence of adverse reaction (14.29％,5/35 cases) in the observation group was lower than that in the control group (38.71％,12/31 cases),and the difference was statistically significant (x2 =5.128,P ＜ 0.05).Conclusion Multi-target combination therapy and traditional cyclophosphamide therapy can effectively control lupus nephritis in children,but the clinical effect of multi-target combination therapy is better and the adverse reaction is less.

Efficacy analysis of sequential blood purification treatment of bee poisoning complicated by multiple organ disfunction syndrome / 中华实用儿科临床杂志

Objective To study the efficacy of sequential blood purification treatment of bee poisoning complicated by multiple organ disfunction syndrome (MODS).Methods The 11 cases of children with bee poisoning and MODS from Wuhan Children's Hospital were treated with sequential blood purification therapy,and they were treated with plasma exchange (PE),hemoperfusion (HP) and sequential continuous renal replacement therapy (CRRT) simultaneously in the early stage,and then were treated with intermittent hemodialysis (IHD) in the remission stage.Different modes of purification treatment were applied in different stages.The trends of liver function,renal function and myocardial enzymes were observed in 11 cases before and after therapy,later a retrospective analysis was performed,and the efficacy of sequential blood purification was studied.Results Ten in 11 cases of children were treated with HP,CRRT and IHD therapy,and among them 6 cases were treated with PE on the first day of admission.One case,the youngest of children admitted to hospital less than 24 hours,died of sudden cardiac arrhythmia due to toxic myocarditis.In ten cases of the children after treatment,their myocardial enzymes returned to normal at first,and then jaundice and hepatic function improved,and renal function gradually improved after 10 days.Two weeks after discharge,through reviewing of the liver and kidney function,myocardial enzymes returned to normal indicators.In review of urine,5 cases were accompanied with microscopic hematuria,3 cases were accompanied with hematuria and proteinuria,and 2 cases were completely normal.The improved cure rate was 91％ (10/11 cases).Conclusions Sequential blood purification treatment is the main and effective means for severe bee poisoning complicated with MODS in children in the early stage.

An experimental research of magnetic resonance tumor targeting imaging with Gd labeled human telomerase reverse transcriptase antisense oligonucleotide (Gd-DOTA-hTERT ASON) / 中国癌症杂志

Background and purpose:Researches had indicated that about over 85%of malignant tumors highly express telomerase activity. So telomerase has become one of the important methods in the research field of tumor diagnosis and treatment. Nowadays, several reports about malignant tumor which over expresses hTERT targeting imaging with radionuclide labeled hTERT ASON had been published. In these reports, high quality of pictures can hardly be acquired because of poor anatomical and spacial resolution in nuclear imaging itself. Accordingly, in this study, we developed a method of detecting human telomerase in vivo with magnetic resonance imaging (MRI) and evaluate its feasibility. Methods:Firstly, Uniformly phosphorothioate-modified human telomerase reverse transcriptase antisense oligonucleotide (hTERT ASON) was labeled with Gd3+ through the bifunctional chelator 1, 4, 7, 10-tetraazacyclododecane-N, N’, N’’, N’’’-tetraacetic acid (DOTA) and iv vitro experiments were performed to characterize the antisense probes (for biodistribution and cellular uptake, 99mTc-DOTA-ASON was used in stead of Gd-DOTA-ASON). Then Gd-DOTA-ASON was injected intraperitoneally in pulmonary adenocarcinoma A375 nude mice tumor-bearing BALB/c for in vivo imaging using 7.0 T Micro MRI periodically, tumors and their surrounding tissues were defined as region of interest (ROI) to calculate the signal to noise ratio (SNR) of tumor to muscle using Gd-DTPA as control. Finally, immunohistochemical analysis of telomerase activity of each xenograft was operated 2 days after imaging. Results:The binding efficiency of Gd-DOTA-ASON reached was as high as 65%(63.2±2.4, n=6). And it can maintain 61%in fresh human serum and normal saline at 37℃over 24 h;A375 cells showed an uptake of 8.5%when incubated with 99mTc-DOTA-ASON;In comparing with DOTA-ASON and Gd-DTPA, cells transected with Gd-DOTA-ASON had higher SI when performed MRI with T1WI. The hTERT-expressing xenografts were obviously enhanced by Gd-DOTA-ASON at 0.5-6 h after injection and the SNR can reach 2.37, whereas obvious enhancement only could be found within 2 h after injection of Gd-DTPA. Both labeled and non-labeled antisense probes can suppress the activity of telomerase of A375 cells either in vitro or in vitro. Conclusion:Our research offers proof that Gd-DOTA-ASON can be used as tumor specific targeting MR probe for diagnosing malignant tumors with high expression of telomerase.

Comparison of short-term effects between interventional embolization treatment and iodine-131 therapy for Graves' hyperthyroidism / 介入放射学杂志

Objective To compare the short-term effects between interventional arterial embolization and iodine-131 therapy in treating Graves'disease.Methods A total of 84 patients with Graves'disease (GD),confirmed by clinical data and laboratory tests,were divided into two groups:interventional group(n=42)receiving arterial embolization and iodine-131 group(n=42)receiving iodine-131 therapy.Before and after the treatment thyroid angiography and SPECT/CT imaging were performed to determine the shape and size of the thyroid,and radioimmunoassay method was used to measure serum levels of FT3,VF4,TSH and TRAb at 3.6 and 12 months after the therapy.The results were compared and statistically analyzed.The occurrence of complications was observed. Results No statistically significant difference in short-term therapeutic effects was found between interventional group and iodine-131 group.The occurrence of early severe complication was much higher in interventional group than that in iodine-131 group,while the occurrence of hypothyroidism was obviously higher in iodine-131 group than that in interventionai group.Conclusion Both interventional embolization and iodine-131 therapy have reliable effect for the treatment of Graves'disease.Iodine-131 therapy may be used in patients who plan to receive initial treatment or in patients who failed to effectively respond to other kinds of therapies,while interventional embolization may be employed in patients who are not able to receive surgery or in patients who have failed to respond to antithyroid drug treatment,or in patients whose iodine-131 intake rate is too low to undergo iodine-131 therapy,especially in patients with refractory and intractable hyperthyroidism. Interventional embolization can be regarded as an alternative treatment for Graves'disease.