Background: Erythrocyte alloantibodies and autoantibodies complicate transfusion. However, the prevalence of erythrocyte alloimmunization and autoimmunization has not been estimated in the Chinese pediatric population. Therefore, we investigated the prevalence of erythrocyte alloimmunization and autoimmunization in the Chinese pediatric population with the aim of developing a reasonable transfusion management policy in children from China. Methods: This study included 30,603 pediatric inpatients who were admitted to three tertiary hospitals in central China from May 2020 to October 2022. Antibody screening was carried out with a three-cell panel by column agglutination technology, and samples with positive screening were analyzed for antibody specificity with a 16-cell identification panel. Clinical details of the patients were collected to identify associations with antibody formation. Results: The alloimmunization rate was 0.55% (169/30,603), and the autoimmunization rate was 0.14% (43/30,603). Alloantibodies comprised 80.09% of the antibodies. The most frequent alloantibodies were anti-M (58.77%), anti-E (9.48%), and anti-P1 (4.27%). Autoantibodies comprised 19.91% of antibodies. Age (p = 0.000), sex (p = 0.016), geographical area (p = 0.000), ABO blood group (p = 0.008), and diagnosis (p = 0.000) were independent risk factors for antibody formation. The risk of antibody formation at the ages of 0-28 days and 1-3 months was zero (odds ratio = 0.000). The antibody distribution was significantly different by age (p = 0.000) and diagnosis (p = 0.000). Conclusion: Repeat pre-transfusion testing for infants less than 4 months of age can be omitted for no risk of antibody formation. MNS system antibodies, especially anti-M, are prominent in younger children, and this decreases with age. Provision of extended phenotype-matched transfusion for Rh system antigens, especially antigen E, is necessary in children to control erythrocyte alloimmunization. The presence of antibodies with high evanescence rates in the pediatric population suggests the pressing need for nationwide shared transfusion records to avoid hemolytic transfusion reactions in children.

Background Subchromosomal deletions and duplications are the leading cause of congenital malformations and mental retardation in children. With the recent clinical application of genomic microarrays in the evaluation of patients with developmental delays and congenital malformations, it has led to the discovery of several new microdeletion and microduplication syndromes. However, there are no published reports involving patients with both microduplications in the 9p21.1-p24.3 region and microdeletions in the 7p22.1-p22.3 region. Case presentation We report an infant with an autosomal abnormality confirmed by conventional karyotype combined with copy number variations sequencing (CNV-seq), showing the patient with an unbalanced translocation. The karyotype of the patient was 46, XX, der (7)t (7;9) (p22; p21) and CNV-seq results showed an approximately 32.34-Mb duplication in 9p21.1-p24.3 (200000-32540000) and an approximately 3.3-Mb deletion in 7p22.2-p22.3 (40000-3340000). Conclusions The patient carried an unbalanced translocation 46, XX, der (7)t (7;9) (p22; p21) derived from her mother. The clinical presentation is closely related to the size and position of the missing and duplicated chromosomes. To our knowledge, the simultaneous occurrence of de novo partial trisomy 9p(9p21.1-p24.3) and partial monosomy 7p (7p22.2-p22.3) has not previously been reported up until now. The present study additionally demonstrated that CNV-seq combined with karyotype is able to reliably detect unbalanced submicroscopic chromosomal aberrations.

Ribonuclease H (RNase H) has been proven to be an attractive biomarker for anti-human immunodeficiency virus (HIV) therapeutics. Herein, a novel label-free electrochemiluminescence (ECL) biosensor is developed for the ultrasensitive detection of RNase H activity by employing DNA/RNA duplex-templated copper (Cu) nanoclusters (NCs). Specifically, interfacial DNA/RNA duplex-templated Cu NCs, as novel excellent ECL luminophores, were successfully synthesized by electroreduction. Meanwhile, the DNA/RNA duplexes can also serve as a target-trigger, exhibiting superior selectivity toward RNase H. Benefitting from the well-designed DNA/RNA duplexes, the label-free ECL biosensor exhibits an admired linear relationship in the concentration range from 3 x 10(-3) U/mL to 20 U/mL with a detection limit of detection of 1.97 x 10(-3) U/mL. Importantly, the detection of RNase H in human serum using the developed ECL biosensor represents satisfactory accuracy, demonstrating that the ECL biosensor possesses tremendous potential for further development in clinical diagnosis.

目的:探讨X染色体异常女性患儿的核型及其临床表型。方法:常规外周血淋巴细胞培养，制备染色体，G显带进行染色体核型分析，必要时用C显带和荧光原位杂交进行验证。结果:71例患儿中，共发现X染色体数目异常29例，占异常核型的40.8%；X染色体结构异常10例，占异常核型的14.1%；嵌合体30例，占异常核型的42.3%；X-常染色体易位2例，占异常核型的2.8%。结论:X染色体异常是导致女性身材矮小、性发育异常及智力低下的重要原因。对此类患儿应进行染色体核型分析以明确病因，及早进行相应的临床干预。同时应开展相关的遗传咨询及产前诊断，以减少缺陷患儿的出生率。

目的 探讨一个腓骨肌萎缩症(charcot-marie-tooth,CMT)家系的临床表现、神经电生理学和基因突变特点.方法 收集家系先证者及其他成员的临床资料,对先证者进行神经电生理学检查和全外显子组基因测序,用Sanger测序技术对先证者及其家系进行突变位点验证.应用计算机软件预测突变位点氨基酸进化保守性和突变可能导致的蛋白质结构和功能变化,分析突变位点的性质.结果 先证者儿童期发病,出现双下肢对称性肌肉无力伴跟腱反射消失及足部畸形,其母亲有类似症状.先证者神经电生理检查示运动和感觉神经纤维脱髓鞘及轴索性改变.基因检测发现先证者和母亲MFN2基因第11个外显子均检出c.1066A＞G(p.T356A)杂合错义突变;先证者姐姐和父亲未检测到该突变.用PolyPhen-2和MutationTaster软件预测该突变为致病性,突变区域序列在不同物种间高度保守.结论 儿童CMT2A2A患者的神经电生理、临床特点、发病机制及相关基因表型均有改变,此可为儿童CMT的临床诊断提供依据.

目的 探讨ALL多探针系统在儿童急性淋巴细胞白血病(acute lymphoblastic leukemia,ALL)诊疗中的应用价值.方法 应用ALL多探针系统(cMYC、P16、E2A、CHIC2/D10Z1/D17Z1、TEL/AMLl、MLL、BCR/ABL1、IGH的DNA探针),进一步检测173例初诊急性淋巴细胞性白血病儿童的分子细胞遗传学改变情况.结果 ALL多探针系统对173例初诊ALL儿童检测中,共检出112例阳性改变,阳性率为64.74％,分别涉及cMYC、P16、E2A、TEL/AMLl、CHIC2/D10Z1/D17Z1、MLL、BCR/ABL1、IGH等8种单一阳性改变109例或合并至少两种阳性改变3例,其中CHIC2/D10Z1/D17Z1超二倍体检测阳性率最高,共76例,阳性率为43.93％,其次是TEL/AMLl阳性,共19例,阳性率为10.98％,另检出8例信号异常,但均未发现阳性异常.结论 儿童ALL患者分子细胞遗传学改变不尽相同,其疾病表现、免疫分型、对治疗的反应及临床预后也各有不同.ALL多探针系统可一次性检出多种与儿童ALL相关的细胞遗传学改变,不仅准确、高效、省力、省时,而且检测效率极高,临床实用性强,非常有利于儿童ALL患者的细胞遗传学诊疗及预后评估,在协助其精准诊疗中具有较强的应用价值.

Objective To explore the genetic basis for a child with clinically suspected 3-methylcrotonyl-coenzyme A carboxylase deficiency (MCCD). Methods Genomic DNA was extracted from peripheral blood samples of the proband and her parents. Whole exome sequencing was used to screen pathogenic variant in the proband. Suspected variant was verified by Sanger sequencing. Impact of the variant on the structure and function of protein product was analyzed by using bioinformatic software. Results Sanger sequencing showed that the proband has carried homozygous missense c.1342G>A (p.Gly448Ala) variant of the MCCC2 gene, for which her mother was a heterozygous carrier. The same variant was not detected in her father. The variant was predicted to be pathogenic by PolyPhen-2 and Mutation Taster software, and the site was highly conserved among various species. Based on the American College of Medical Genetics and Genomics standards and guidelines, the c.1342G>A (p.Gly448Ala) variant of MCCC2 gene was predicted to be likely pathogenic(PM2+PP2-PP5). Conclusion The homozygous missense variant of the MCCC2 gene c.1342G>A (p.Gly448Ala) probably underlay the molecular pathogenesis of the proband. Genetic testing has confirmed the clinical diagnosis.

目的:通过对本地区491例隐匿性阴茎患儿进行细胞遗传学分析,以了解此类患儿性染色体异常情况及所伴染色体病特点,为临床诊治提供资料.方法:回顾性分析本院2017年5月—2019年7月临床诊断为隐匿性阴茎的491例患儿的细胞遗传学资料.结果:在491例隐匿性阴茎患儿中,共发现性染色体异常9例,异常检出率为1.83％.其中克氏综合征患儿共7例,占总体异常检出率的77.78％,检出超雄综合征患儿1例,占总体异常检出率的11.11％,另检出男性特纳氏综合征患儿1例,占总体异常检出率的11.11％.结论:隐匿性阴茎是一种先天发育异常及常见的阴茎畸形性疾病,部分此类患儿可合并有性染色体异常,最多见的是克氏综合征,细胞遗传学手段能够明确隐匿性阴茎患儿是否伴有染色体病.对于伴有染色体病的此类患儿,不仅要及时采用手术治疗隐匿性阴茎,也应及早发现并诊疗染色体病,另外遗传咨询及健康教育对于伴染色体病的隐匿性阴茎患儿的生长发育及预后具有重要意义.

OBJECTIVE:To analyze the causes of positive irregular antibody screening test and incompatibility of cross matching in one patient with autoimmune hemolytic anemia complicated with neonatal hemolytic disease, and to accurately identify the type of antibodies in patients, and to select a reasonable strategy for blood transfusion.METHODS:One children was enrolled, blood group positive and reverse typing, Rh typing, direct anti-human globulin test, free test, dispersal test and cross matching test were carried out by test tube method and microcolumn gel card; irregular antibodies were identified by the reaction of DTT treatment and untreated panel cells with patients' plasma.RESULTS:The blood group of the patient was RhD positive B and irregular antibody screening positive, while the blood group of the mother was RhD positive O and irregular anti-screening negative, the result showed that the anti-LW detected in the plasma of the patient was autoantibody and ABO neonatal hemolytic disease (ABO-HDN) was present. Both O type RhD positive washing RBCs and B type RhD negative RBCs were transfused effectively.CONCLUSION:Irregular antibodies in patients are anti-LW antibodies, and transfusion of homotype RhD negative suspended erythrocytes after the exclusion of ABO-HDN shows a better effect.

目的 探讨染色体核型分析联合荧光原位杂交(FISH)检测在儿童急性髓系白血病M2中的诊疗意义.方法 选择25例AML-M2儿童骨髓标本应用染色体G显带进行核型分析,并同时应用荧光原位杂交技术(FISH)检测AML1/ETO融合基因.结果 在25例AML-M2儿童中,核型分析检出t(8;21)染色体易位17例(68％),包括除典型t(8;21)及其它染色体异常,如复杂易位、缺失、额外染色体等,核型正常5例(20％),其中4例为复诊儿童,3例(12％)标本因细胞生长不良,制片质量差,无法分析;25例AML-M2儿童同时做FISH,检出阳性20例(80％),包括17例t(8;21)及3例无分裂相的标本,4例复诊儿童均为阴性,检出信号异常1例(4％),但无阳性改变.结论 急性髓系白血病M2患儿特异性t(8;21)/AML1-ETO融合基因阳性改变较易检出,FISH技术检测患儿易位的敏感性较高于染色体核型分析.FISH检测不仅可以提示融合基因是否阳性,也可提示特定基因是否异常.染色体核型分析可同时提示相关染色体是否易位及其余染色体有无畸变.因此,两种分析方法联合应用可多方面提示AML-M2儿童的疾病信息,在AML-M2的诊治及预后中具有重要的应用价值.

The abnormal change of thrombin concentration is closely related with various diseases. By using proximity binding-induced DNA network assembled by nonlinear hybridization chain reaction (HCR), we here developed an enzyme-free and ultrasensitive electrochemical strategy for analyzing thrombin in human serums. The binding of the two different aptamers to a thrombin enhanced the local concentration of the aptamers and facilitates the formation of an intact strand. Subsequently, the formed strand caused nonlinear HCR, forming DNA network. The DNA network bound to the capture probes (CP)-modified electrode, producing electrochemical signal of MB. Benefiting from the proximity binding-induced DNA network, the proposed biosensor exhibited a wide linear range from 1.0 pM to 1.0 nM with a low detection limit of 0.56 pM and high specificity. More importantly, the developed enzyme-free electrochemical biosensor has been used to detect thrombin in serum samples with good performance. This makes such a strategy possess potential promise in the application of clinical diagnosis.

目的 研究β-地中海贫血患者长期红细胞输注中应用RhCcEe抗原配合性输血策略的效果.方法 选取2018年5月至2019年8月郑州大学附属儿童医院收治的86例β-地中海贫血患者作为研究对象,采用系统随机化法均分为研究组(ABO、RhC、c、D、E、e抗原相合性或相容性输血)和对照组(ABO、RhD相合性输血).观察两组输血12周后的临床疗效,比较两组输血前及输血12周后的血液学指标[血红蛋白(Hb)、血细胞比容(HCT)]、心脏功能[脑钠肽(BNP)、心肌肌钙蛋白I(cTnI)],观察两组不良反应发生情况及不规则抗体产生情况.结果 输血12周后,两组治疗总有效率比较,差异无统计学意义(P＞0.05).输血12周后,研究组Hb、HCT水平均较输血前显著上升(P＜0.05),输血前及输血12周后,两组Hb、HCT水平比较,差异无统计学意义(P＞0.05).输血12周后,两组BNP、cTn I水平均较输血前显著下降,研究组明显低于对照组(P＜0.05).研究组不良反应发生率明显低于对照组(P＜0.05).两组不规则抗体阳性率比较,差异无统计学意义(P＞0.05).结论 RhCcEe抗原配合性输血策略在β-地中海贫血患者长期红细胞输注中应用效果良好,能够有效地减少患者输血不良反应,改善心脏功能.

OBJECTIVE:To explore the genetic basis of a pedigree affected with peroneal muscular atrophy.METHODS:Neuroelectrophysiological examination and whole exome sequencing were carried out for the proband, a six-year-and-ten-month-old boy. Suspected variant was verified in his family members through Sanger sequencing. Bioinformatic analysis was carried to predict the conservation of amino acid sequence and impact of the variant on the protein structure and function.RESULTS:Electrophysiological examination showed demyelination and axonal changes of motor and sensory nerve fibers. A heterozygous missense c.1066A>G (p. Thr356Ala) variant was found in exon 11 of the MFN2 gene in the proband and his mother, but not in his sister and father. Bioinformatic analysis using PolyPhen-2 and Mutation Taster software predicted the variant to be pathogenic, and that the sequence of variation site was highly conserved among various species. Based no the American College of Medical Genetics and Genomics standards and guidelines, the c.1066A>G (p. Thr356Ala) variant of MFN2 gene was predicted to be likely pathogenic (PS1+ PM2+ PP3+ PP4).CONCLUSION:The heterozygous missense c.1066A>G (p.Thr356Ala) variant of the MFN2 gene probably underlay the disease in the proband, and the results have enabled genetic counseling and prenatal diagnosis for this family.

目的:通过比较轻症及重症手足口患儿血清α-INF、β-INF及γ-INF指标,探讨其变化特点及临床意义.方法:手足口患儿88例,其中轻症患儿44例、重症患儿44例,检测两组患儿血清α-INF、β-INF及γ-INF水平变化.结果:两组患儿血清α-INF、β-INF、γ-INF变化,差异均具有统计学意义(P＜0.05).结论:手足口病患儿血清干扰素水平的变化可能与手足口病的病情进展有着密不可分的联系.

Objective To explore the effects of immunoglobulin combined with acupuncture therapy on limb function and prognosis and rehabilitation in children with hand-foot-and-mouth disease and limb flaccid paralysis.Methods A total of 48 children with hand-foot-and-mouth disease and limb flaccid paralysis admitted to our hospital from January,2016 to January,2019 were selected,and divided into an observation group and a control group by random number table method,with 24 cases in each group.The two groups were given conventional therapies,such as exercise therapy,neurotrophic drugs and hyperbaric oxygen therapy;in addition,the observation group was given immunoglobulin and acupuncture therapy.The clinical efficacy,recovery times of neurological signs (muscle tension recovery time and tendon reflex recovery time),D and E functional areas of lower extremity motor function [Gross Motor Function Measure (GMFM)],and cytokines [anti-GM1 antibody,intedeukin-6 (IL-6),interleukin-8 (IL-8),tumor necrosis factor-α (TNF-α)] before and 5 d after the treatment were compared between the two groups.Results The total effective rate was higher in the observation group than in the control group (P ＜ 0.05).The muscle tension recovery time and tendon reflex recovery time were lower in the observation group than in the control group (both P ＜ 0.05).5 d after the treatment,the scores of D and E areas of GMFM scale were higher than those before the treatment in the two groups,and were higher in the observation group than in the control group (all P ＜ 0.05).5 d after the treatment,the anti-GM1 antibody,IL-6,IL-8 and TNF-α were lower than those before the treatment in the two groups,and were lower in the observation group than in the control group (all P ＜ 0.05).Conclusion Immunoglobulin combined with acupuncture therapy in children with hand-foot-and-mouth disease complicated with limb flaccid paralysis can help the recovery of limb function and improve the prognosis.Its immunoregulatory effects of anti-GM1 antibody,IL-6,IL-8,TNF-α and other cytokines are the possible mechanisms.