Complex heterozygous mutations in hereditary spherocytosis:A case report

BACKGROUND The aim of this study was to investigate the complex heterozygous mutations of ANK1 and SPTA1 in the same individual and improve our understanding of hereditary spherocytosis(HS)in children.We also hope to promote the application of gene detection technology in children with HS,with the goals of identifying more related gene mutations,supporting the acquisition of improved molecular genetic information to further reveal the pathogenesis of HS in children,and providing important guidance for the diagnosis,treatment,and prevention of HS in children.CASE SUMMARY A 1-year and 5-month-old patient presented jaundice during the neonatal period,mild anemia 8 months later,splenic enlargement at 1 year and 5 months,and brittle red blood cell permeability.Genetic testing was performed on the patient,their parents,and sister.Swiss Model software was used to predict the protein structure of complex heterozygous mutations in ANK1 and SPTA1.Genetic testing revealed that the patient harbored a new mutation in the ANK1 gene from the father and a mutation in the SPTA1 gene from the mother.Combined with the clinical symptoms of the children,it is suggested that the newly discovered complex heterozygous mutations of ANK1 and SPTA1 may be the cause,providing important guidance for revealing the pathogenesis,diagnosis,treatment,and promotion of gene detection technology in children with HS.CONCLUSION This case involves an unreported complex heterozygous mutation of ANK1 and SPTA1,which provides a reference for exploring HS.

Polycystic kidney and hepatic disease 1 gene mutations in von Meyenburg complexes: Case report

Von Meyenburg complexes(VMCs) are a rare type of ductal plate malformation. We herein report two Chinese families with VMCs, and the suspicious gene mutation of this disease. Proband A was a 62-year-old woman with abnormal echographic presentation of the liver. She received magnetic resonance imaging(MRI) examination and liver biopsy, and the results showed she had VMCs. Histologically proved hepatocellular carcinoma was found 1 year after the diagnosis of VMCs. Proband B was a 57-year-old woman with intrahepatic diffuselesions displayed by abdominal ultrasonography. Her final diagnoses were VMCs, congenital hepatic fibrosis, and hepatitis B surface e antigen-negative chronic hepatitis B after a series of examinations. Then, all the family members of both proband A and proband B were screened for VMCs by MRI or ultrasonography. The results showed that four of the 11 family members from two families, including two males and two females, were diagnosed with VMCs. DNA samples were extracted from the peripheral blood of those 11 individuals of two VMCs pedigrees and subjected to polymerase chain reaction amplification of the polycystic kidney and hepatic disease 1(PKHD1) gene. Two different mutation loci were identified. Heterozygous mutations located in exon 32(c.4280 delG, p.Gly1427 ValfsX 6) in family A and exon 28(c.3118 C>T, p.Arg1040 Ter) in family B were detected. We speculate that PKHD1 gene mutations may be responsible for the development of VMCs.

Effect of the mutation of carotenoids on the dynamics of energy transfer in light-harvesting complexes （LH2） from Rhodobacter sphaeroides 601 at room temperature

Energy transfers in two kinds of peripheral light-harvesting complexes （LH2） of Rhodobacter sphaeroides （RS） 601 are studied by using femtosecond pump^probe spectroscopy with tunable laser wavelength at room temperature. These two complexes are native LH2 （RS601） and green carotenoid mutated LH2 （GM309）. The obtained results demonstrate that, compared with spheroidenes with ten conjugated double bonds in native RS601, carotenoid in GM309 containing neurosporenes with nine conjugated double bonds can lead to a reduction in energy transfer rate in the B800-to-B850 band and the disturbance in the energy relaxation processes within the excitonic B850 band.

Complete mitochondrial DNA sequence analysis in two southern Chinese pedigrees with Leber hereditary optic neuropathy revealed secondary mutations along with the primary mutation

AIM: To investigate mitochondrial factors associated with Leber hereditary optic neuropathy (LHON) through complete sequencing and analysis of the mitochondrial genome of Chinese patients with this disease. METHODS: Two unrelated southern Chinese families with LHON and 10 matched healthy controls were recruited, and their entire mitochondrial DNA (mtDNA) was amplified and sequenced with the universal M13 primer. Then DNA sequence analysis and variation identification were performed by DNAssist and Chromas 2 software and compared with authoritative databases such as Mitomap. RESULTS: Mutational analysis of mtDNA in these two Chinese pedigrees revealed one common LHON-associated mutation, G11778A (Arg -> His), in the MT-ND4 gene. In addition, there were two secondary mutations in Pedigree 1: C34971 (Ala -> Val), and C3571T (Leu -> Phe) in the MT-ND1 gene, which have not been reported; and two secondary mutations occurred in Pedigree 2: A10398G (Thr -> Ala) in the MT-ND3 gene, and T14502C (Ile -> Val) in the MT-ND6 gene. Three polymorphisms, A73G, G94A and A263G in the mtDNA control region, were also found. CONCLUSION: Our study confirmed that the known MT-ND4* G11778A mutation is the most significant cause of LHON. The C3497T and C3571T mutations in Pedigree 1 were also both at hot-spots of MT-ND1; they may affect the respiratory chain in coordination with the primary mutation G11778A. In Pedigree 2, the two secondary mutations A10398G of MT-ND3 and T14502C of MT-ND6 may influence mitochondrial respiratory complex I, leading to the mitochondrial respiratory chain dysfunction which results in optic atrophy together with G11778A. Therefore, not only the common primary LHON mutation is responsible for the visual atrophy, but other secondary mtDNA mutations should also be considered when giving genetic counseling.

Efficacy of EGFR Tyrosine Kinase Inhibitors in Non-small Cell Lung Cancer Patients Harboring Different Types of EGFR Mutations:A Retrospective Analysis

With the development of molecular pathology, many types of epidermal growth factor receptor（EGFR） mutations have been identified. The efficacy of EGFR tyrosine kinase inhibitors（EGFR-TKIs） in non-small cell lung cancer（NSCLC） patients with different types of EGFR mutations, especially in patients with single rare mutations or complex mutations（co-occurrence of two or more different mutations）, has not been fully understood. This study aimed to examine the efficacy of EGFR-TKIs in NSCLC patients with different types of EGFR mutations. Clinical data of 809 NSCLC patients who harbored different types of EGFR mutations and treated from January 2012 to October 2016 at Renmin Hospital and Zhongnan Hospital, Wuhan, were retrospectively reviewed. The clinical characteristics of these patients and the efficacy of EGFR-TKIs were analyzed. Among these patients, 377 patients had only the EGFR del-19 mutation, 362 patients the EGFR L858R mutation in exon 21, 33 patients single rare mutations and 37 patients complex mutations. Among these 809 patients, 239 patients were treated with EGFR-TKIs. In all the 239 patients, the disease control rate（DCR） was 93.7% with two patients（0.2%） achieving complete response（CR）, the median progression free survival（PFS） was 13.0 months（95% confidence interval [CI], 11.6–14.4 months）, and the median overall survival（OS） was 55.0 months（95% CI, 26.3–83.7 months）. Subgroup analysis revealed that the DCR in patients harboring single rare or complex mutations of EGFR was significantly lower than in those with del-19 or L858 R mutation（P〈0.001）. Patients with classic mutations（del-19 and/or L858 R mutations） demonstrated longer PFS（P〈0.001） and OS（P=0.017） than those with uncommon mutations（single rare and/or complex mutations）. Furthermore, the patients with single rare mutations had shorter median OS than in those with other mutations. Multivariate Cox regression analysis identified that the type of EGFR mutations was an independent risk factor for PFS（hazard ratio [HR]=0.308, 95% CI, 0.191–0.494, P〈0.001） and OS（HR=0.221, 95% CI, 0.101–0.480, P〈0.001）. The results suggest that the single rare or complex EGFR mutations confer inferior efficacy of EGFR-TKIs treatment to the classic mutations. The prognosis of the single rare EGFR mutations is depressing. EGFR-TKIs may be not a good choice for NSCLC patients with single rare mutations of EGFR. Further studies in these patients with uncommon mutations（especially for the patients with single rare mutations） are needed to determine a better precision treatment.

Presenilin mutations and their impact on neuronal differentiation in Alzheimer’s disease

The presenilin genes(PSEN1 and PSEN2)are mainly responsible for causing early-onset familial Alzheimer’s disease,harboring~300 causative mutations,and representing~90%of all mutations associated with a very aggressive disease form.Presenilin 1 is the catalytic core of theγ-secretase complex that conducts the intramembranous proteolytic excision of multiple transmembrane proteins like the amyloid precursor protein,Notch-1,N-and E-cadherin,LRP,Syndecan,Delta,Jagged,CD44,ErbB4,and Nectin1a.Presenilin 1 plays an essential role in neural progenitor maintenance,neurogenesis,neurite outgrowth,synaptic function,neuronal function,myelination,and plasticity.Therefore,an imbalance caused by mutations in presenilin 1/γ-secretase might cause aberrant signaling,synaptic dysfunction,memory impairment,and increased Aβ42/Aβ40 ratio,contributing to neurodegeneration during the initial stages of Alzheimer’s disease pathogenesis.This review focuses on the neuronal differentiation dysregulation mediated by PSEN1 mutations in Alzheimer’s disease.Furthermore,we emphasize the importance of Alzheimer’s disease-induced pluripotent stem cells models in analyzing PSEN1 mutations implication over the early stages of the Alzheimer’s disease pathogenesis throughout neuronal differentiation impairment.

Exploring Biocomplexity in Cancer: A Comprehensive Review

Living objects have complex internal and external interactions. The complexity is regulated and controlled by homeostasis, which is the balance of multiple opposing influences. The environmental effects finally guide the self-organized structure. The living systems are open, dynamic structures performing random, stationary, stochastic, self-organizing processes. The self-organizing procedure is defined by the spatial-temporal fractal structure, which is self-similar both in space and time. The system’s complexity appears in its energetics, which tries the most efficient use of the available energies;for that, it organizes various well-connected networks. The controller of environmental relations is the Darwinian selection on a long-time scale. The energetics optimize the healthy processes tuned to the highest efficacy and minimal loss (minimalization of the entropy production). The organism is built up by morphogenetic rules and develops various networks from the genetic level to the organism. The networks have intensive crosstalk and form a balance in the Nash equilibrium, which is the homeostatic state in healthy conditions. Homeostasis may be described as a Nash equilibrium, which ensures energy distribution in a “democratic” way regarding the functions of the parts in the complete system. Cancer radically changes the network system in the organism. Cancer is a network disease. Deviation from healthy networking appears at every level, from genetic (molecular) to cells, tissues, organs, and organisms. The strong proliferation of malignant tissue is the origin of most of the life-threatening processes. The weak side of cancer development is the change of complex information networking in the system, being vulnerable to immune attacks. Cancer cells are masters of adaptation and evade immune surveillance. This hiding process can be broken by electromagnetic nonionizing radiation, for which the malignant structure has no adaptation strategy. Our objective is to review the different sides of living complexity and use the knowledge to fight against cancer.

SWI/SNF染色质重塑复合物在肿瘤耐药中的作用

肿瘤耐药是化疗和靶向药物治疗面临的重要问题,它是一个涉及染色质重塑的复杂过程。SWI/SNF是肿瘤发生中研究最多的ATP依赖性染色质重塑复合物之一,在染色质结构稳定性、基因表达和翻译后修饰的协调过程中起着重要作用,但其调控肿瘤耐药的机制尚未被系统梳理。SWI/SNF按其亚基组成可分为BAF、PBAF和ncBAF 3种,这3种亚型均包含ATP酶催化亚基、核心亚基和调节亚基,可以通过调控染色质结构来控制基因表达。SWI/SNF复合物亚基的改变是调控肿瘤耐药和进展的重要因素之一,充分了解该复合物在肿瘤耐药中的分子机制可以改善癌症的治疗。本文总结了SWI/SNF复合物的组成、引起肿瘤耐药的突变或异常表达亚基类型、主要的耐药机制以及肿瘤耐药的克服方法等,为临床上SWI/SNF亚基突变或异常表达引起的肿瘤预后不良的诊断和治疗提供了思路。

新发TSC2基因位点突变致儿童结节性硬化症并色素脱斑相关癫痫

结节性硬化症(tuberous sclerosis complex,TSC)是一种常染色体显性遗传的神经皮肤综合征,以累及多个器官系统为特点。TSC1和TSC2是TSC两个主要的致病基因,二者中任一基因的突变可导致蛋白质结构变化从而导致功能改变,最终表现为TSC的各种临床表型。目前,已有多个TSC相关的TSC2和TSC1位点突变被发现。然而,临床接诊过程中,我们收治了1例尚未见报道的TSC2基因c.4569+1G>T杂合突变相关的癫痫发作伴色素脱斑的儿童TSC,在此予以报道,以期为TSC相关疾病的临床诊断及研究提供线索。

薄胼胝体和“猞猁耳征”——遗传性痉挛性截瘫11型一家系报告

报道1例MRI影像表现为典型薄胼胝体(thin corpus callosum,TCC)和“猞猁耳征(ears-of-the-lynx sign)”的遗传性痉挛性截瘫11型(spastic paraplegia 11,SPG11)病例。患者为13岁男孩,主要表现走路不稳容易跌倒,症状逐渐加重伴双手书写不灵、学习能力下降1年;父母无血缘关系。患儿头颅MRI表现为胼胝体变薄,双侧脑室前角旁白质T2、Flair轴位对称性高信号,全外显子组测序检测到SPG11基因NM_025137:c.2073delT和c.257+5G>A两处杂合突变分别来自父母,先证者最终诊断为SPG11,并给予对症治疗。脑MRI影像表现TCC和“猞猁耳征”对诊断SPG11具有高度敏感性和特异性,对于有痉挛性截瘫表现的患者要考虑到SPG11的可能。

基于降维粒子群的大电网断面功率极限计算方法

随着电力系统的发展,系统运行方式日益复杂。为了提高电力系统运行安全水平,提出一种基于降维粒子群的大电网断面功率极限计算方法。首先综合考虑大电网静态安全稳定约束和暂态稳定约束,建立断面功率极限计算模型;然后,提出了基于功率灵敏度的降维粒子群算法,根据功率灵敏度和聚合系数对可调节发电机组进行分群处理,利用降维粒子群算法计算得到了极限功率、发电机组功率调节量以及极限功率制约因素,实现了大电网断面功率极限的快速寻优。最后在我国某地区2128节点电网和我国某跨区12643节点联网中验证了该方法的有效性和正确性。

结节性硬化症三例及基因突变分析

目的:明确3例结节性硬化症(tuberous sclerosis complex, TSC)患者基因突变位点。方法:对3例患者血液样本进行全外显子基因检测,对患者3进行一代测序验证。结果:患者1存在结节性硬化1型(TSC1)基因杂合插入变异TSC1:NM_000368.5:exon10:c.989dupT:p.S331Efs^(*)10;患者2和患者3发现存在结节性硬化2型(TSC2)基因的杂合突变,分别是:TSC2:NM_000548.5:exon22:c.2481_2486 del:p.V828_K829 del和TSC2:NM_000548.5:exon5:c.348delG:p.V118Sfs^(*)64;患者3父母未检出该变异。结论:患者2和患者3的2个突变在OMIM中未查询到相应记录,患者3推测为新发变异或父母一方存在生殖细胞嵌合,丰富了该疾病的突变位点谱。

1例伴TSC2嵌合突变的结节性硬化症并多发血管平滑肌脂肪瘤的诊断及治疗

目的探讨结节性硬化症(TSC)相关肝血管平滑肌脂肪瘤(AML)、肾AML患者的诊断、治疗及病程中基因诊断的局限性和多学科诊疗的重要性。方法基于1例患者详尽的临床资料,依托多学科会诊,对病程中多次基因检测结果进行深入分析,明确诊断后给予药物治疗并评价疗效。患者为31岁男性,患有肝、肾多发不典型AML 12年,同时检查发现鼻部及口周血管纤维瘤、臀部鲨革斑、头部MRI提示皮层发育不良、牙釉质缺损史等多系统表现,多年来仅以反复AML切除术为主要治疗方法。为进一步确认患者的基因突变特征及明确诊断,进行了深入基因分析。结果经深入基因分析发现,外周血和肿瘤组织中分别存在突变丰度4.04%和10.38%的TSC2 c.2353C>T(p.Gln785*)突变,考虑在胚胎发育阶段出现生殖细胞嵌合突变可能。诊断为AML合并TSC。给予患者mTOR抑制剂依维莫司治疗1年,经复查患者肾多发AML病灶较前显著缩小,疗效达到部分缓解。结论AML患者需警惕是否合并TSC。诊断TSC时,低丰度的胚系突变需深入分析。mTOR抑制剂可作为TSC-AML患者治疗的选择。

A Chinese Tuberous Sclerosis Complex Family and a Novel Tuberous Sclerosis Complex-2 Mutation

Tuberous sclerosis complex （TSC） is a relatively common autosomal dominant genetic disorder affecting l/14,000-1/6000 Western populations.The incidence of TSC in Chinese population is still unknown although case reports of Chinese TSC patients were documented. The main clinical features of TSC include seizures,mental retardation,and the development ofhamartomas in multiple organs such as the skin,brain,lung,heart,and kidney.Indeed,the disease virtually manifests in every organ. Two causative genes for TSC,TSC 1 gene on chromosome 9q34 and TSC2 gene on chromosome16p13,have been identified in 1997 and 1993 respectively.Approximately,70% of cases of TSC are de novo mutations. Chinese TSC patients are more likely to have TSC2 missense and frame shift mutations.Here,we record one Chinese TSC family and it is novel frame shift mutation of TSC2.

多梳抑制性去泛素化酶复合物的结构与功能及其在血液肿瘤发生发展中的作用

多梳抑制性去泛素化酶(polycomb repressive deubiquitinase,PR-DUB)复合物是多梳蛋白家族的成员之一,通过调节组蛋白修饰参与染色体的表观遗传修饰。多梳抑制性复合物1(polycomb repressive complex 1,PRC1)和PR-DUB复合物通过对H2AK119Ub泛素化与去泛素化修饰调控平衡,保护活性基因免受异常沉默,去泛素化功能与促进基因活化和建立转录允许的染色质状态有关,除此之外还激活增强子并促进双链断裂处DNA损伤修复。附加性梳样1(additional sex comb-like1,ASXL1)作为表观遗传支架组装染色质修饰复合物和转录因子参与表观遗传调控。BRCA1相关蛋白1(BRCA1-associated protein 1,BAP1)作为去泛素化酶去除底物的泛素化修饰。PR-DUB复合物由核心二聚体和其他辅助因子组成,BAP1与ASXL1形成核心二聚体,其他亚基相互作用调节PR-DUB复合物靶向和功能。ASXL1和BAP1是与PR-DUB复合物去泛素化功能最相关的2个亚基,ASXL1的DEUBAD结构域激活BAP1发挥去泛素化作用水解H2AK119Ub1。了解ASXL1和BAP1的结构以及相互作用机制对研究PR-DUB复合物特异性去泛素化作用的机制至关重要。在人类中,PR-DUB复合物成分的突变经常引起多种血液肿瘤。ASXL 1基因突变常导致蛋白质翻译提前结束,大部分是由于C末端PHD结构域缺失导致。目前认为,PR-DUB复合物中突变的ASXL1或BAP1、表观遗传因子以及Akt/mTOR等靶点或信号通路相互作用是促进血液肿瘤发生发展的可能机制。这对于针对潜在的治疗靶点研究开发新的特异性靶向治疗药物至关重要。本文将介绍PR-DUB复合物的结构与功能、作用机制及其在血液肿瘤疾病中的发生,重点就ASXL1和BAP1进行综述,并系统总结了潜在的靶向治疗药物,以期为PR-DUB复合物在血液疾病防治中的研究提供科学参考。

复杂山区铁路隧道勘察设计工程质量风险识别及耦合演化分析

铁路隧道工程勘察设计阶段工作对整个工程的全生命周期质量具有重要影响。然而,山区地质条件复杂、自然灾害频发等特点给铁路隧道勘察设计带来极大挑战,而且铁路工程质量风险构成复杂,风险事件的发生通常是多风险耦合作用的结果。目前关于铁路隧道的研究多聚焦于隧道的关键技术、施工安全、绿色性能等方面,鲜有立足勘察设计阶段的山区铁路隧道质量风险耦合研究。为从勘察设计阶段提高隧道工程质量目标的成效,提出基于突变理论的质量风险耦合效应分析模型。首先从内部诱因(人员、技术、管理)和外部诱因(地质、社会、自然)2个维度辨识影响复杂山区铁路隧道勘察设计的质量风险源,并结合改进重要性-绩效分析法(IPA)对风险因子进行层级归类;其次分析多风险因素耦合的形成机理,基于突变理论从耦合度和稳定性2个方面分析铁路隧道勘察设计工程质量风险耦合效应,并构建尖点突变模型进行风险耦合路径演化分析;最后以某复杂山区铁路隧道为例,利用提出的方法进行验证,确定其风险耦合等级为Ⅲ级,存在高风险的耦合程度,质量风险需要警惕,但整体系统处于稳定状态。该研究可以为复杂山区铁路隧道勘察设计工程质量风险决策提供理论支撑。

A Novel Mutation of PRKAR1A Caused Carney Complex in a Chinese Patient

Primary pigmented nodular adrenocortical disease （PPNAD） causes adrenocorticotropic hormone （ACTH）-independent Cushing＇s syndrome （CS）,which is the most frequent endocrine manifestation of Carney complex （CNC）.[1] In the disease process of PPNAD,both presadrenal glands are involved and feature small brown-black nodules separated by the atrophic adrenal cortex.

Bayesian Analysis of Complex Mutations in HBV, HCV,and HIV Studies

In this article,we aim to provide a thorough review of the Bayesian-inference-based methods applied to Hepatitis B Virus(HBV),Hepatitis C Virus(HCV),and Human Immunodeficiency Virus(HIV)studies with a focus on the detection of the viral mutations and various problems which are correlated to these mutations.It is particularly difficult to detect and interpret these interacting mutation patterns,but by using Bayesian statistical modeling,it provides a groundbreaking opportunity to solve these problems.Here we summarize Bayesian-based statistical approaches,including the Bayesian Variable Partition(BVP)model,Bayesian Network(BN),and the Recursive Model Selection(RMS)procedure,which are designed to detect the mutations and to make further inferences to the comprehensive dependence structure among the interactions.BVP,BN,and RMS in which Markov Chain Monte Carlo(MCMC)methods are used have been widely applied in HBV,HCV,and HIV studies in the recent years.We also provide a summary of the Bayesian methods’applications toward these viruses’studies,where several important and useful results have been discovered.We envisage the applications of more modified Bayesian methods to other infectious diseases and cancer cells that will be following with critical medical results before long.

Malignancy of renal angiomyolipoma from tuberous sclerosis complex with TSC2 mutation

To the Editor:Tuberous sclerosis complex (TSC),with the birth incidence of 1:6000,[1] is an autosomal dominant inherited,multi-system disorder characterized by cellular hyperplasia and tissue dysplasia,among which,renal angiomyolipoma (AML) is one common comorbidity.However,malignancy of renal AML is rare.Herein,we shared a case of malignancy of renal AML from TSC in a young man.

Recombination and mutation shape variations in the major histocompatibility complex

The major histocompatibility complex(MHC)is closely associated with numerous diseases,but its high degree of polymorphism complicates the discovery of disease-associated variants.In principle,recombination and de novo mutations are two critical factors responsible for MHC polymorphisms.However,direct evidence for this hypothesis is lacking.Here,we report the generation of fine-scale MHC recombination and de novo mutation maps of~5 Mb by deep sequencing(>100×)of the MHC genome for 17 MHC recombination and 30 non-recombination Han Chinese families(a total of 190 individuals).Recombination hotspots and Han-specific breakpoints are located in close proximity at haplotype block boundaries.The average MHC de novo mutation rate is higher than the genome-wide de novo mutation rate,particularly in MHC recombinant individuals.Notably,mutation and recombination generated polymorphisms are located within and outside linkage disequilibrium regions of the MHC,respectively,and evolution of the MHC locus was mainly controlled by positive selection.These findings provide insights on the evolutionary causes of the MHC diversity and may facilitate the identification of disease-associated genetic variants.