Potential protein–phenotype correlation in three lipopolysaccharideresponsive beige-like anchor protein-deficient patients

BACKGROUND Patients with lipopolysaccharide(LPS)-responsive beige-like anchor protein(LRBA)deficiency have a variety of clinical symptoms,but there is no apparent genotype–phenotype correlation,and patients carrying the same mutations may have different phenotypes.Therefore,it is not easy for doctors to make a decision regarding hematopoietic stem cell transplantation(HSCT)for LRBA-deficient patients.We hypothesized that there may be a protein–phenotype correlation to indicate HSCT for LRBA-deficient patients.AIM To report on three Chinese LRBA-deficient patients and determine the correlation between residual protein expression and disease phenotypes.METHODS Clinical data of three Chinese LRBA-deficient patients were collected,and protein levels were detected by Western blot analysis.In addition,LRBA mutation information of another 83 previously reported patients was summarized.RESULTS All the major clinical findings indicated enteropathy,but patients 1 and 3 presented with more severe symptoms than patient 2.Endoscopy and histology indicated nonspecific colitis for patients 1 and 3 but Crohn's disease-like colitis for patient 2.Compound heterozygous mutations in LRBA were found in patient 1,and homozygous mutations in LRBA were found in patient 2 and patient 3.Only patient 2 responded well to traditional immunosuppressive treatment.Residual expression of the LRBA protein in patients 1 and 3 was very low,but in patient 2,a more than 0.5-fold in expression of the LRBA protein was found compared to that in the control.After HSCT,patient 1 had increased LRBA protein expression.We summarized the genetic information of 86 patients,and the mutations in patients 1 and 3 were novel mutations.CONCLUSION We described three Chinese LRBA-deficient patients,two of whom carried novel mutations.These patients had no genotype-phenotype correlations,but their residual LRBA protein expression might be associated with disease outcome and could be an indicator for HSCT.

Transfer of the extensor indicis proprius branch of posterior interosseous nerve to reconstruct ulnar nerve and median nerve injured proximally:an anatomical study

Proximal or middle lesions of the ulnar or median nerves are responsible for extensive loss of hand motor function.This occurs even when the most meticulous microsurgical techniques or nerve grafts are used.Previous studies had proposed that nerve transfer was more effective than nerve grafting for nerve repair.Our hypothesis is that transfer of the posterior interosseous nerve,which contains mainly motor fibers,to the ulnar or median nerve can innervate the intrinsic muscles of hands.The present study sought to investigate the feasibility of reconstruction of the deep branch of the ulnar nerve and the thenar branch of median nerve by transferring the extensor indicis proprius branch of the posterior interosseous nerve obtained from adult cadavers.The results suggested that the extensor indicis proprius branch of the posterior interosseous nerve had approximately similar diameters and number of fascicles and myelinated nerve fibers to those of the deep branch of ulnar nerve and the thenar branch of the median nerve.These confirm the feasibility of extensor indicis proprius branch of posterior interosseous nerve transfer for reconstruction of the deep branch of the ulnar nerve and the thenar branch of median nerve.This procedure could be a novel and effective method for the functional recovery of the intrinsic muscles of hands after ulnar nerve or median nerve injury.

Risk stratification of hemodynamically significant patent ductus arteriosus by clinical and genetic factors

Background Hemodynamically significant patent ductus arteriosus(hsPDA)is associated with increased comorbidities in neonates.Early evaluation of hsPDA risk is critical to implement individualized intervention.The aim of the study was to provide a powerful reference for the early identification of high-risk hsPDA population and early treatment decisions.Methods We enrolled infants who were diagnosed with PDA and performed exome sequencing.The collapsing analyses were used to find the risk gene set(RGS)of hsPDA for model construction.The credibility of RGS was proven by RNA sequencing.Multivariate logistic regression was performed to establish models combining clinical and genetic features.The models were evaluated by area under the receiver operating curve(AUC)and decision curve analysis(DCA).Results In this retrospective cohort study of 2199 PDA patients,549(25.0%)infants were diagnosed with hsPDA.The model[all clinical characteristics selected by least absolute shrinkage and selection operator regression(all CCs)]based on six clinical variables was acquired within three days of life,including gestational age(GA),respiratory distress syndrome(RDS),the lowest platelet count,invasive mechanical ventilation,and positive inotropic and vasoactive drugs.It has an AUC of 0.790[95%confidence interval(CI)=0.749–0.832],while the simplified model(basic clinical characteristic model)including GA and RDS has an AUC of 0.753(95%CI=0.706–0.799).There was a certain consistency between RGS and differentially expressed genes of the ductus arteriosus in mice.The AUC of the models was improved by RGS,and the improvement was significant(all CCs vs.all CCs+RGS:0.790 vs.0.817,P<0.001).DCA demonstrated that all models were clinically useful.Conclusions Models based on clinical factors were developed to accurately stratify the risk of hsPDA in the first three days of life.Genetic features might further improve the model performance.

Nickel-catalyzed reductive coupling reaction of monofluoroalkyl triflates with alkyl carboxylic acids toward the synthesis of α-alkyl-α-fluoro-alkylketones

The synthesis methods ofα-fluoro-arylketones were well-established through electrophilic/nucleophilic fluorination and transition metal catalyzed cross-coupling.However,due to the site selectivity and substrate restriction,only a few cases have been developed to affordα-alkyl-α-fluoro-alkylketones.Herein,we report a general and efficient method of preparing diverseα-alkyl-α-fluoro-alkylketones via nickelcatalyzed reductive coupling reaction of monofluoroalkyl triflates with low-cost industrial raw material alkyl carboxylic acids.These transformations demonstrate high efficiency,mild conditions,and excellent functional group compatibility.This strategy provides a general and efficient method for the synthesis ofα-alkyl-α-fluoro-alkylketones。

Application of next generation sequencing in the screening of monogenic diseases in China,2021:a consensus among Chinese newborn screening experts

Newborn screening(NBS)refers to a maternal and newborn healthcare technology,in which special examinations of congenital and genetic diseases that could seriously impact the health of newborns,are implemented during the neonatal period to provide early diagnosis and treatment[1].With a history of more than 60 years,NBS has advanced greatly due to technological progress resulting in significant improvement in the number of diseases covered by NBS and in screening efficiency[2-7].

Identification of eight novel mutations in 11 Chinese patients with maple syrup urine disease

Background Maple syrup urine disease(MSUD)is an autosomal recessive inherited disorder that affects the degradation of branched-chain amino acids and is associated with acute and chronic brain dysfunction.This study presents 11 new patients with MSUD and describes the clinical characteristics and gene mutations reported in Chinese individuals.Methods During 2011–2018,11 pedaitric patients with MSUD from 11 Chinese families were analyzed based on clinical characteristics and mass spectrometry,with confirmation via gene sequencing.Novel mutations affecting protein function were predicted with Mutation-Taster,PolyPhen-2,CADD and SIFT software.3D models of the mutated proteins were generated by using the SWISS-MODEL online server,and the models were visualized in PyMOL.The characteristics and gene mutations in patients with MSUD were analyzed retrospectively.Results Seventeen mutations in the BCKDHA,BCKDHB and DBT genes were found,8 of which are novel:c.55C>/T,c.349C>T,c.565C>T,c.808G>A,c.859C>G,and c.1270dupC in BCKDHA;c.275-2A>G in BCKDHB;and c.1291C>T in DBT.Eight patients died.Two patients had severe mental retardation and were physically handicapped.One patient with the intermediate type had relatively good prognosis,with mild psychomotor retardation and adiposity.Four mothers underwent amniocentesis for prenatal diagnosis during their second pregnancy;two fetuses were wild type,and two were carriers of one heterozygous mutation.Conclusions Eight novel mutations were associated with MSUD in Chinese patients.Prenatal diagnosis was successfully performed by genetic analysis.Mutations in the BCKDHB gene were found in the majority of Chinese patients with MSUD.

Visible-light-induced,autopromoted nickel-catalyzed threecomponent arylsulfonation of 1,3-enynes and mechanistic insights

A light-induced,nickel-catalyzed three-component arylsulfonation of 1,3-enynes in the absence of photocatalyst is reported.This methodology exhibited mild conditions,broad scope and high efficiency,and its synthetic utility has been demonstrated by a concise total synthesis of sulfone-containing drug molecule.Detailed mechanistic studies indicated that this light induced nickel catalysis is autopromoted by in situ produced allene,which plays a key role as co-ligand in the photoactive excited state Ni(Ⅰ)species for the LMCT process.The detailed elucidation of this light-induced nickel catalytic cycle may shed some lights on the exploitation of new catalytic activity and establishment of novel methods.

Ligand-Promoted,Enantioconvergent Synthesis of Aliphatic Alkanes Bearing Trifluoromethylated Stereocenters via Hydrotrifluoroalkylation of Unactivated Alkenes

While drug chirality remains one of the most important themes in medicinal chemistry and fluorine-containing drugs have comprised large portion of global pharmaceutical market,the synthesis of fluorine-containing compounds featuring a trifluoromethylated stereogenic carbon center is still underdeveloped.Recent strategy of enantioconvergent cross-coupling of trifluoromethylated alkyl halide has significantly updated previous toolbox that is dependent on additional functional group to achieve desired bond formation and highly enantioselective control.However,evident limitations were present in specific coupling candidate(sp?carbon)and/or prerequisite heteroatom in trifluoroalkyl source for inducing effective stereoconvergent differentiation.Leveraging novel design of sterically hindered bisoxazoline ligand and drastic increase of molecular complexity of in situ generated sp3 coupling partner via metal-hydride atom transfer,herein,we report a nickel-catalyzed,highly enantioselective hydrotrifluoroalkylation of unactivated alkenes for diverse synthesis of enantioenriched aliphatic alkanes featuring trifluoromehylated stereogenic carbon.Excellent stereochemical control(mostly>95:5 er),high catalytic reactivity,mild conditions,and broad substrate scope(40+examples),enable convenient late-stage asymmetric trifluoroalkylation of various biologically active complexmolecules.

Effects of Bifidobacterium supplementation on intestinal microbiota composition and the immune response in healthy infants

Background:Intestinal microbiotas are thought to be the most important source of maturational stimuli to the development of the immune system.However,few studies have focused on the development of T helper(Th)1 immune response and antibody response to vaccinations in healthy infants,especially in a large cohort.Through this randomized,double-blind control trial,we investigated the effects of Bifidobacterium longum BB536(BB536)supplementation on intestinal microbiota composition and the immune response in term infants.Methods:In total,300 healthy newborns were recruited,randomized and fed formula either supplemented with BB536 or with no supplementation.Stool samples were analyzed at months 2,4 and 11.The representative cytokine for Th1[interferon-γ(IFN-γ)]and Th2[interleukin-4(IL-4)]secretion cells were measured using enzyme-linked immunospot assay at 4 and 7 months of age.The antibody response to vaccines was measured at months 7 and 11.Results:A total of 264 infants completed the study.The amount of bifidobacteria and the bifidobacteria/Enterobacteriaceae ratio(B/E)were signifi cantly higher in the BB536 supplementation group at months 2 and 4.The number of IFN-γsecretion cells and the ratio of IFN-γ/IL-4 secretion cells were increased in the BB536 supplementation group at 7 months.Moreover,the higher value of B/E in the early stages seems to be related to the increased Th1 response.No difference was observed between groups in the antibody response after vaccination.Conclusions:BB536 has positive effects on establishing a healthy intestinal microbiota early in life,and it also plays an important role in improving the Th1 immune response.

Genetic etiologies associated with infantile hydrocephalus in a Chinese infantile cohort

Background Infantile hydrocephalus(IHC)is commonly related to other central nervous system diseases,which may have adverse effects on prognosis.The causes of IHC are heterogeneous,and the genetic etiologies are not fully understood.This study aimed to analyze the genetic etiologies of an IHC cohort.Methods The data for 110 IHC patients who had received exome sequencing at the Clinical Genetic Center of the Children's Hospital of Fudan University between 2016 and 2019 were reviewed and analyzed retrospectively.An exome-wide association analysis(EWAS)was performed within this cohort using IHC as the study phenotype.Results Of the 110 IHC patients,a pathogenic or likely pathogenic variant was identified in 16(15%)patients,spanning 13 genes.The genes were mainly associated with metabolic disorders,brain abnormalities,and genetic syndromes.IHC patients who had unclear clinical etiology were more likely to possess a genetic etiology.Based on previous studies and on our EWAS results,ZEB1,SBF2,and GNAI2 were over-represented among IHC patients and might alfect the signaling pathways involved in IHC formation.Conclusions Our study showed heterogeneous genetic etiologies in an IHC cohort.It is essential to perform genetic testing on IHC patients who have unclear clinical etiology,and genes associated with metabolic disorders,brain abnormalities,and genetic syndromes should he noted.In addition,when aiming to discover IHC susceptibility genes,genes that might influence the signaling pathways involved in IHC formation should be prioritized.

Selective 1,4-arylsulfonation of 1,3-enynes via photoredox/nickel dual catalysis

A photoredox/nickel-catalyzed selective 1,4-arylsulfonation of 1,3-enynes to access structurally diverse sulfone-containing allenes has been established.This radical cascade transformation featured easy manipulation,mild conditions,low catalyst loading,broad substrate scope,and large-scale synthesis.The preliminary mechanistic studies indicated a possible radical-relay process enabled by the radical capture of nickel(0)species.

Responsible genes in children with primary vesicoureteral reflux: findings from the Chinese Children Genetic Kidney Disease Database

Background Primary vesicoureteral reflux(VUR)is a common congenital anomaly of the kidney and urinary tract(CAKUT)in childhood.The present study identified the possible genetic contributions to primary VUR in children.Methods Patients with primary VUR were enrolled and analysed based on a national multi-center registration network(Chinese Children Genetic Kidney Disease Database,CCGKDD)that covered 23 different provinces/regions in China from 2014 to 2019.Genetic causes were sought using whole-exome sequencing(WES)or targeted-exome sequencing.Results A total of 379 unrelated patients(male:female 219:160)with primary VUR were recruited.Sixty-four(16.9%)children had extrarenal manifestations,and 165(43.5%)patients showed the coexistence of other CAKUT phenotypes.Eighty-eight patient(23.2%)exhibited impaired renal function at their last visit,and 18 of them(20.5%)developed ESRD at the median age of 7.0(IQR 0.9–11.4)years.A monogenic cause was identified in 28 patients(7.39%).These genes included PAX2(n=4),TNXB(n=3),GATA3(n=3),SLIT2(n=3),ROBO2(n=2),TBX18(n=2),and the other 11 genes(one gene for each patient).There was a significant difference in the rate of gene mutations between patients with or without extrarenal complications(14.1%vs.6%,P=0.035).The frequency of genetic abnormality was not statistically significant based on the coexistence of another CAKUT(9.6%vs.5.6%,P=0.139,Chi-square test)and the grade of reflux(9.4%vs.6.7%,P=0.429).Kaplan–Meier survival curve showed that the presence of genetic mutations did affect renal survival(Log-rank test,P=0.01).PAX2 mutation carriers(HR 5.1,95%CI 1.3–20.0;P=0.02)and TNXB mutation carriers(HR 20.3,95%CI 2.4–168.7;P=0.01)were associated with increased risk of progression to ESRD.Conclusions PAX2,TNXB,GATA3 and SLIT2 were the main underlying monogenic causes and accounted for up to 46.4%of monogenic VUR.Extrarenal complications and renal function were significantly related to the findings of genetic factors in children with primary VUR.Like other types of CAKUT,several genes may be responsible for isolated VUR.

Secondary genomic findings in the 2020 China Neonatal Genomes Project participants

Background During next generation sequencing(NGS)data interpretation in critically ill newborns,there is a potential for recognizing and reporting secondary findings(SFs).Early awareness of SFs may provide clues for disease prevention.In this study,we assessed the frequency of SFs in the China Neonatal Genomes Project(CNGP)participants.Methods A total of 2020 clinical exome sequencing(CES)datasets were screened for variants from a list of 59 genes recommended by the American College of Medical Genetics and Genomics(ACMG)for secondary findings reporting v2.0(ACMG SF v2.0).Identified variants were classified according to the evidence-based guidelines reached by a joint consensus of the ACMG and the Association for Molecular Pathology(AMP).Results Among the 2020 CES datasets,we identified 23 ACMG-reportable genes in 61 individuals,resulting in an overall frequency of SFs at 3.02%.A total of 53 unique variants were identified,including 35 pathogenic and 18 likely pathogenic variants.The common disease categories of SFs associated were cardiovascular and cancer disease.The SF results affected the medical management and follow-up strategy in 49(80.3%)patients.Conclusions We presented the frequency of SFs and their impact on clinical management strategies in CNGP participants.Our study demonstrated that SFs have important practical value in disease prevention and intervention at an early stage.