SRY基因在性发育异常诊断中的应用

目的为SRY在临床分子诊断中的应用提供参考,并对性发育异常机制进行探讨。方法通过对32例性发育异常患者和男性不育症患者进行SRY基因的检查分析,应用聚合酶链式反应（PCR）对性发育异常患者和男性不育症患者进行SRY检测。结果检出成功率为93.75%（30/32）,异常率为28.13%（9/32）,其中女性SRY扩增阳性率为9.38%（3/32）,男性SRY扩增阴性率为18.75%（6/32）。结论SRY基因是性别分化的关键基因,SRY基因的缺失或突变是造成性发育异常的主要原因,研究也表明人类的性别决定和分化还有其他相关基因的参与。对性发育异常患者进行SRY基因检测,有利于了解该类患者的遗传学病因,为其诊断和治疗提供科学依据。     

社区护理在发育性髋关节不良儿童中的应用

目的分析社区护理在发育性髋关节不良儿童中的应用效果。方法收集我院2012年6月~2013年6月诊治的发育性髋关节不良儿童患儿80例作为研究对象,以抛硬币的方式分为试验组与对照组,每组患儿各40例。对照组患儿采用常规护理模式,试验组患儿在对照组的基础上实施社区护理,对两组患儿的护理效果进行分析对比。结果研究结果显示,试验组患儿的并发症发生率明显低于对照组(<0.05),患儿及其家长的满意度明显高于对照组(<0.05)。两组患儿护理前生活质量评分比较无明显差异(>0.05),经护理后均得到明显改善(<0.05),且试验组改善水平明显优于对照组(<0.05)。结论社区护理在发育性髋关节不良儿童中具有良好的应用效果,能有效降低并发症发生率,提高患者及其家属的满意度,促进生活质量改善,值得在临床应用上推广。     

髋臼中心定位工具在Crowe Ⅳ型发育性髋关节发育不良关节置换术中的应用

目的 研究髋臼中心定位工具在Crowe Ⅳ型发育性髋关节发育不良关节置换术中的可行性及安全性。方法 回顾性分析2020年1月至2022年1月在新疆维吾尔自治区伊犁哈萨克自治州新华医院行全髋关节置换术的37例（44髋）发育性髋关节脱位患者,分为两组：A组20例（24髋）,参考定位工具,B组17例（20髋）,参考髋臼横韧带,比较两组患者的手术时间、切口大小、术中出血量、下地时间、Harris髋关节评分、双侧肢体长度差,术后臼杯外展角、前倾角,以及术后假体旋转中心与解剖旋转中心的水平、垂直距离差值。结果 所有的患者术后未出现脱位、感染、深静脉血栓形成、假体松动等。两组患者的手术时间、术中出血量、切口大小、下地时间、Harris评分、双侧肢体长度差比较,差异无统计学意义（P＞0.05）。对比两组臼杯外展角、前倾角,差异无统计学意义（P＞0.05）;旋转中心垂直距离A组为（21.54±2.32） mm、B组为（22.40±2.23） mm,水平距离A组为（29.42±2.45） mm、B组为（29.85±2.92） mm,重建髋臼的旋转中心距离及其与解剖旋转中心距离差进行组间比较,差异无统计学意义（P＞0.05）。结论 髋臼中心定位工具可以在Crowe Ⅳ型发育性髋关节发育不良全髋关节置换术中给术者提供髋臼中心点的参考标志,有一定的临床使用价值。     

荧光原位杂交技术结合染色体核型分析在儿童性染色体异常导致性发育异常中的应用

目的回顾性分析性染色体异常导致的性发育异常(DSD)患儿的性染色体遗传学分布及临床表现。方法分析2013年1月至2022年3月就诊于河南省儿童医院郑州儿童医院14 857例存在矮小、隐睾、尿道下裂、隐匿性阴茎、生长发育落后等DSD患儿的临床资料, 应用荧光原位杂交技术(FISH)和染色体核型分析遗传学病因。结果共检出423例由性染色体异常导致的DSD患儿, 阳性率检出约为2.85%(423/14 857)。符合特纳综合征(TS)特征性染色体为XO及其嵌合体患儿有327例(77.30%)。其中, 社会性别为女性患儿325例, 临床表现以身材矮小为主, 社会性别为男性患儿2例, 主要表现为身材矮小、隐睾、尿道下裂。符合克氏综合征(KS)特点性染色体为XXY及其嵌合体患儿有62例(14.66%), 主要临床表现为隐睾、隐匿性阴茎、尿道下裂等。性染色体为XO/XY嵌合体19例(4.49%), 女性患儿(11例)临床表现均为矮小, 男性患儿(8例)临床表现均有尿道下裂, 6例在尿道下裂基础上合并患有隐睾、隐匿性阴茎、睾丸扭转等。其他类型15例(3.55%), 包含性染色体为XYY及其嵌合体患儿9...     

低深度全基因组测序拷贝数变异分析技术在性发育异常患儿诊断中的应用价值

目的探讨低深度全基因组测序拷贝数变异分析(CNV-seq)技术在性发育异常(DSD)患儿诊断中的应用价值。方法纳入2019年10月至2020年10月至郑州大学第一附属医院就诊的5例身材矮小或外阴发育异常的DSD患儿。在外周血染色体核型分析、全外显子组测序(WES)、SRY基因检测的基础上, 进行CNV-seq检测以明确病因。结果患儿1和2的社会性别为女性, 染色体核型均为46, XY, WES结果为阴性, CNV-seq结果分别为46, XY, +Y(1.4)和46, XY, -Y(0.75)。其余3例患儿均携带可疑的Y染色体, 综合分析发现其核型分别为45, X[60]/46, X, del(Y)(q11.221)[40]、45, X, 16qh+[76]/46, X, del(Y)(q11.222), 16qh+[24]和45, X[75]/46, XY[25]。结论联合运用CNV-seq等分子遗传学技术明确了46, XY DSD患儿的Y染色体拷贝数变异及45, X/46, XY DSD患儿可疑Y染色体的性质, 为其临床诊疗提供了重要的依据。     

Repercussions of surgically assisted maxillary expansion on nose width and position of septum and inferior nasal conchae

The aim of the present study was to assess the clinical and radiographic repercussions of surgically assisted maxillary expansion on the septum, nasal cavity and nasal conchae. The sample was made up of 15 patients with skeletal maturity (9 females and 6 males between 16 and 45 years of age) and maxillary transverse deficiency. Assessments were performed through anterior rhinoscopy and frontal cephalometric radiographs on three occasions: (T0) preoperative period, (T1) locking of the expander and (T2) six months following the locking procedure. An increase was observed in the basal portion of the pyriform aperture and distances between the lateral wall of the basal portion of the pyriform aperture and the septum. The radiographic exam revealed that the nasal septum did not undergo any statistically significant change in its position. Moreover, no significant changes in the position of the nasal septum or nasal conchae were detected throughout the three evaluation times. The results suggest that surgically assisted maxillary expansion is capable of widening the basal portion of the pyriform aperture, with little repercussion on the anterior position of the nasal septum and inferior nasal conchae.     

RUNX2-related metaphyseal dysplasia with maxillary hypoplasia: A rare skeletal disorder resembling SFRP4-related Pyle disease

Metaphyseal dysplasia with maxillary hypoplasia with or without brachydactyly (MDMHB) is an ultra-rare skeletal dysplasia caused by heterozygous intragenic RUNX2 duplications, comprising either exons 3 to 5 or exons 3 to 6 of RUNX2. In this study, we describe a 14-year-old Belgian boy with metaphyseal dysplasia with maxillary hypoplasia but without brachydactyly. Clinical and radiographic examination revealed mild facial dysmorphism, dental anomalies, enlarged clavicles, genua valga and metaphyseal flaring and thin cortices with an osteoporotic skeletal appearance. Exome sequencing led to the identification of a de novo heterozygous tandem duplication within RUNX2, encompassing exons 3 to 7. This duplication is larger than the ones previously reported in MDMHB cases since it extends into the C-terminal activation domain of RUNX2. We review previously reported cases with MDMHB and highlight the resemblance of this disorder with Pyle disease, which may be explained by intersecting molecular pathways between RUNX2 and sFRP4. This study expands our knowledge on the genotypic and phenotypic characteristics of MDMHB and the role of RUNX2 in rare bone disorders.     

A homozygous ROR2 variant in a family with atypical Robinow syndrome and tetramelic transverse deficiency of autopods

We present five members of a consanguineous Pakistani kinship with the most severe familial tetramelic transverse autopod deficiency reported to date and additionally having some of the common autosomal recessive Robinow syndrome-1 (RRS1) features including short stature, short neck, severe vertebral anomalies of kyphoscoliosis, hemivertebrae, fusion of thoracic vertebrae, broad forehead, and dental crowding. We mapped the locus of this atypical RRS and detected homozygous 8-nucleotide deletion c.1353_1360del (p.(Met452Alafs*4)) in ROR2, the gene responsible for RRS1. We did not find any other variant shared by all affected individuals that could possibly act as a modifier of limb defect. Autopods are affected in RRS1, but severe autopod deficiency is not a characteristic feature. Over 30 biallelic variants dispersed throughout the gene are known in ROR2-related RS, with no genotype–phenotype correlation for specific RRS1 features. Considering together with the sporadic case homozygous for variant p.(Arg442*) and the case homozygous for p.(Arg441Thrfs*16) in a family where heterozygous members have brachydactyly type B1, we propose that homozygous truncating variants that originate at residues 441–452 can cause severe autopod reduction anomalies, suggesting some genotype–phenotype correlation for this particular phenotype.     

A simple reliable assay for myeloperoxidase activity in mixed neutrophil-eosinophil cell suspensions: application to detection of myeloperoxidase deficiency

Quantitation of myeloperoxidase (MPO) activity by guaiacol peroxidation (GP) assay is profoundly affected by the peroxidase present in eosinophils (EPO) that contaminate the granulocyte suspensions. Inclusion of 3-amino-1,2,4-triazole (AMT) in the GP assay permits quantitation of MPO activity in mixed neutrophil-eosinophil suspension because of the differential inhibition of EPO and MPO by AMT. Results show that: (1) the peroxidase activity of eosinophil-free granulocyte suspensions is not appreciably affected by AMT; (2) in the presence of AMT the peroxidase activities of granulocyte preparations containing different numbers of eosinophils are similar on a neutrophil basis, regardless of the number of eosinophils and correspond with the activity of eosinophil-free granulocyte suspensions; (3) AMT almost completely inhibits the activity of partially purified EPO, only slightly affecting the catalytic activity of partially purified MPO; (4) AMT completely inhibits the residual peroxidase activity of granulocyte suspensions from MPO-deficient subjects contributed by contaminating eosinophils. The GP assay in the presence of AMT was used to study the pattern of hereditary transmission of MPO deficiency. The genealogy derived on the basis of this assay was compatible with an autosomal recessive inheritance, in agreement with previously reported results, while no definite pattern of inheritance could be established by use of the GP assay without AMT. We suggest that the GP assay supplemented with AMT is the method of choice for detection of MPO deficiency, particularly partial deficiency.     

Triosephosphate isomerase deficiency: biochemical and molecular genetic analysis for prenatal diagnosis

Inherited deficiency of the glycolytic enzyme triosephosphate isomerase leads to a multisystem disorder characterized by progressive neuromuscular dysfunction, chronic nonspherocytic hemolytic anemia and increased susceptibility to severe infections. Most patients die within the first 6 years. We examined a family with severe triosephosphate isomerase deficiency. The 1-year-old index patient suffered from hemolytic anemia, neuromuscular impairment and pneumonias, with the necessity of intermittent mechanical ventilation. Triosephosphate isomerase activity in erythrocytes was reduced to about 20% of normal. Heat stability of the enzyme was strongly reduced; concentration of the physiological substrate, dihydroxyacetone phosphate was increased 20-fold due to the metabolic block. Direct sequencing of the triosephosphate isomerase gene revealed homozygosity for the formerly described GAGGA C -mutation changing 104 GluAsp. During a 2nd pregnancy we examined a cord blood sample obtained in the 19th gestational week. The biochemical data on enzyme activity, heat stability of the enzyme and concentration of dihydroxyacetone phosphate were in the normal range. The molecular genetic analysis confirmed the presence of the normal triosephosphate isomerase alleles. Pregnancy was continued, resulting in the delivery of an unaffected, healthy newborn.     

Fructose-1,6-diphosphatase deficiency and glyceroluria: one possible etiology for GIS

Fructose-1,6-diphosphatase (FDPase) deficiency is characterized by episodes of lactic acidemia, hypoglycemia, and ketonuria. Liver biopsy and subsequent enzyme analysis most reliably make the diagnosis. Review of the literature reveals 85 cases. Glycerol intolerance syndrome (GIS) is less well defined. There are only a handful of cases reported. We describe a patient with FDPase deficiency and significant glyceroluria and propose that GIS may be caused by partial deficiency of FDPase.     

Allogeneic hematopoietic cell transplantation for primary immune deficiency diseases: current status and critical needs

Allogeneic hematopoietic cell transplantation (HCT) has been used for 40 years to ameliorate or cure primary immune deficiency (PID) diseases, including severe combined immunodeficiency (SCID) and non-SCID PID. There is a critical need for evaluation of the North American experience of different HCT approaches for these diseases to identify best practices and plan future investigative clinical trials. Our survey of incidence and prevalence of PID in North American practice sites indicates that such studies are feasible. A conference of experts in HCT treatment of PID has recommended (1) a comprehensive cross-sectional and retrospective analysis of HCT survivors with SCID; (2) a prospective study of patients with SCID receiving HCT, with comparable baseline and follow-up testing across participating centers; (3) a pilot study of newborn screening for SCID to identify affected infants before compromise by infection; and (4) studies of the natural history of disease in patients who do or do not receive HCT for the non-SCID diseases of Wiskott-Aldrich syndrome and chronic granulomatous disease. To accomplish these goals, collaboration by a consortium of institutions in North America is proposed. Participation of immunologists and HCT physicians having interest in PID and experts in laboratory methods, clinical outcomes assessment, databases, and analysis will be required for the success of these studies.     

Autocrine and immune cell‐derived BDNF in human skeletal muscle: implications for myogenesis and tissue regeneration

The neurotrophin system has a role in skeletal muscle biology. Conditional depletion of BDNF in mouse muscle precursor cells alters myogenesis and regeneration in vivo. However, the expression, localization and function of BDNF in human skeletal muscle tissue is not known, so the relevance of the rodent findings for human muscle are unknown. Here we address this by combining ex vivo histological investigations on human biopsies with in vitro analyses of human primary myocytes. We found that BDNF was expressed by precursor and differentiated cells both in vitro and in vivo. Differential analysis of BDNF receptors showed expression of p75NTR and not of TrkB in myocytes, suggesting that the BDNF–p75NTR axis is predominant in human skeletal muscle cells. Several in vitro functional experiments demonstrated that BDNF gene silencing or protein blockade in myoblast cultures hampered myogenesis. Finally, histological investigations of inflammatory myopathy biopsies revealed that infiltrating immune cells localized preferentially near p75NTR‐positive regenerating fibres and that they produced BDNF. In conclusion, BDNF is an autocrine factor for skeletal muscle cells and may regulate human myogenesis. Furthermore, the preferential localization of BDNF‐producing immune cells near p75NTR‐positive regenerating myofibres suggests that immune cell‐derived BDNF may sustain tissue repair in inflamed muscle. Copyright © 2013 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.     

FiNuTyper: Design and validation of an automated deep learning-based platform for simultaneous fiber and nucleus type analysis in human skeletal muscle

Aim

While manual quantification is still considered the gold standard for skeletal muscle histological analysis, it is time-consuming and prone to investigator bias. To address this challenge, we assembled an automated image analysis pipeline, FiNuTyper (Fi ber and Nu cleus Typer ).

Methods

We integrated recently developed deep learning-based image segmentation methods, optimized for unbiased evaluation of fresh and postmortem human skeletal muscle, and utilized SERCA1 and SERCA2 as type-specific myonucleus and myofiber markers after validating them against the traditional use of MyHC isoforms.

Results

Parameters including cross-sectional area, myonuclei per fiber, myonuclear domain, central myonuclei per fiber, and grouped myofiber ratio were determined in a fiber-type-specific manner, revealing that a large degree of sex- and muscle-related heterogeneity could be detected using the pipeline. Our platform was also tested on pathological muscle tissue (ALS and IBM) and adapted for the detection of other resident cell types (leucocytes, satellite cells, capillary endothelium).

Conclusion

In summary, we present an automated image analysis tool for the simultaneous quantification of myofiber and myonuclear types, to characterize the composition and structure of healthy and diseased human skeletal muscle.