Pregnant women with systemic lupus erythematosus (SLE) are at high risk of preeclampsia (PE). However, overlapping clinical features between the two conditions make it challenging to detect superimposed PE. Failure to promptly recognize PE significantly increases the risk of adverse pregnancy outcomes. In recent years, complete blood count–derived inflammatory indicators (e.g., platelet-to-lymphocyte ratio) have emerged as readily available, low-cost biomarkers with clinical utility. This study aimed to assess the discriminative capacity of these derived inflammatory indicators, measured within one week before delivery, for identifying superimposed PE in pregnant patients with SLE. This retrospective single-center study included 157 pregnant women with SLE treated at Nanfang Hospital of Southern Medical University from April 2010 to October 2024. Clinical and laboratory data, including complete blood count, liver function tests, and coagulation parameters, were collected. Univariate and multivariate logistic regression were performed to identify risk factors for PE and construct combined models. Model performance was assessed using receiver operating characteristic (ROC) curves, Hosmer–Lemeshow test, calibration plots, and the integrated discrimination improvement (IDI) and net reclassification improvement (NRI) indices. Internal validation used the bootstrap method, and sensitivity analysis used E-values. Among the study patients, 35 developed PE. Univariate logistic regression analysis showed significant differences in monocyte-to-lymphocyte ratio (MLR) and platelet-to-lymphocyte ratio (PLR) between groups (P < 0.05). Multivariable logistic regression identified that lower MLR (OR = 0.223, 95

BACKGROUND:It has traditionally been considered that mother-infant ABO incompatibility only causes mild haemolytic disease of the newborn (HDN). However, this view is inconsistent with clinical practice, and large-scale population-based data are lacking to investigate its effects on neonates. METHODS:Differences in hospitalisation rates and incidence rates of neonatal hyperbilirubinaemia (NHB) and anaemia among 47 679 Chinese liveborn neonates with different mother-infant ABO combinations, differences in the incidence of ABO-incompatible HDN (ABO-HDN) among neonates with O-B versus O-A mother-infant ABO incompatibility, and the contributions of ABO-HDN to the development of NHB and neonatal anaemia were analysed. RESULTS:Of the 47 679 liveborn neonates, neonates with mother-infant ABO incompatibility had higher rates of hospitalisation and incidence of NHB and anaemia. The hierarchy of the risk of mother-infant ABO incompatibility to the neonate was O-B > O-A > non-O-A/O-B incompatibility. Among neonates with O-B and O-A mother-infant ABO incompatibility, the ABO-HDN incidence rates were 15.27% (513/3359) and 11.33% (417/3680), respectively (95% CI 1.41 (1.23 to 1.62)), and the severe ABO-HDN incidence rates were 2.05% (69/3359) and 1.14% (42/3680), respectively (95% CI 1.82 (1.23 to 2.67)). Among the 7039 neonates with O-A/O-B mother-infant ABO incompatibility, ABO-HDN was an independent aetiological factor in 41.11% (666/1620) of the neonates with NHB, 70.27% (52/74) of the neonates with severe NHB, 42.34% (163/385) of the neonates with anaemia and 18.28% (17/93) of the neonates with severe anaemia. CONCLUSIONS:Mother-infant ABO incompatibility often leads to severe HDN and is a dominant cause of NHB and neonatal anaemia, leading to significantly higher neonatal hospitalisation rates.

BACKGROUND:Polycystic ovary syndrome (PCOS) is a metabolic disorder affecting women of reproductive age, and its etiology remains unclear. Therefore, it is crucial to identify biomarkers of the metabolic disturbances in PCOS. METHODS:A total of eight clinical PCOS samples and control group samples were analyzed using data-independent acquisition (DIA) proteomics. Clinical data were used to identify protein biomarkers, and enzyme-linked immunosorbent assay (ELISA) validation was performed on 27 PCOS and 23 control samples. RESULTS:In the PCOS samples, a total of 114 differentially expressed proteins were identified, with 37 upregulated and 77 downregulated. Further biofunctional analysis using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways revealed two differentially expressed proteins, lactate dehydrogenase A (LDHA) and triosephosphate isomerase 1 (TPI1), both of which were significantly upregulated in clinical PCOS samples. LDHA and TPI1 are involved in the glycolysis/gluconeogenesis metabolic pathway. Finally, ELISA was used to validate the high expression of LDHA and TPI1 in PCOS patients. CONCLUSION:DIA proteomics effectively identifies PCOS biomarkers. LDHA and TPI1 may serve as diagnostic biomarkers and could exert effects through glycolytic pathways.

Background: Abnormal concentrations of maternal thyroid hormones are risk factors for certain obstetrical complications. However, the influence induced by different types of maternal thyroid dysfunction on obstetrical complications and outcomes remains controversial. This study aimed to systematically evaluate the prevalence of distinct thyroid dysfunction subtypes in pregnant women and their specific associations with adverse obstetric outcomes, thereby clarifying clinical management priorities. Methods: In a retrospective cohort study, a total of 17,219 pregnant women underwent a thyroid function test, including thyroid stimulating hormone (TSH) and free tetraiodothyronine (fT4). All participants were divided into seven groups based on their blood test results, and their pregnancy outcomes were followed up. The isolated hypothyroxinemia group was divided into two cohorts, depending on whether the patients received levothyroxine. Complications during pregnancy and the outcomes were observed and analyzed in both cohorts. Results: A total of 2621 (15.22%) women were identified with an abnormal thyroid function, including 1150 with subclinical hypothyroidism, 562 with gestational transient thyrotoxicosis, 419 with subclinical hyperthyroidism, 336 with isolated hypothyroxinemia, 78 with hyperthyroidism, and 76 with hypothyroidism. After adjusting for maternal characteristics, no significant associations were found between specific hyperthyroidism groups and the risk of pregnancy complications. However, mothers with overt hypothyroidism had nearly a 3-fold increased risk of developing postpartum hemorrhage (odds ratio (OR): 2.76; 95% confidence interval (95% CI): 1.19–6.38; p = 0.018). Subclinical hypothyroidism was associated with an increased risk of premature membrane rupture (OR: 1.44; 95% CI: 1.25–1.64; p < 0.001) and therapeutic abortion related to fetal anomalies (OR: 2.05; 95% CI: 1.13–3.74; p = 0.019). Additionally, both subclinical hypothyroidism, overt hypothyroidism, and isolated hypothyroxinemia were linked to more than a 2-fold increase in the risk of preeclampsia. Mothers with subclinical hypothyroidism exhibited a lower risk for gestational diabetes mellitus (OR: 0.67; 95% CI: 0.57–0.79; p < 0.001), while those with isolated hypothyroxinemia had approximately a 1.5-fold increased risk for gestational diabetes mellitus (OR: 1.41; 95% CI: 1.11–1.80; p = 0.005). There were no significant differences in outcomes between those receiving levothyroxine treatment in the isolated hypothyroxinemia group and those who did not. Conclusions: Our results showed a high incidence of thyroid dysfunction in pregnant women, with subclinical hypothyroidism being the most common, followed by gestational transient thyrotoxicosis. In general, pregnant women with hypothyroidism presented with a high risk of complications during pregnancy. Isolated hypothyroxinemia in pregnant women is concerning, and levothyroxine treatment did not improve pregnancy outcomes and obstetrical complications.

Objectives: We aim to establish a predictive model for recurrent preeclampsia. Methods: A retrospective review of medical records from three hospitals between 2010 and 2021 was conducted. The study included women who had two consecutive singleton deliveries at the same hospital, with the first delivery complicated by preeclampsia. A multivariable logistic regression model was constructed using a training cohort, and subsequently cross-validated and tested using an independent cohort. The model's performance was assessed in terms of discrimination and calibration, and its clinical utility was evaluated using decision curve analysis (DCA). Results: Among 296 405 deliveries, 694 women met the inclusion criteria, with 151 (21.8%) experiencing recurrent preeclampsia. The predictive model incorporated 10 risk factors from previous preeclampsia, including gestational weeks with elevated blood pressure, gestational diabetes mellitus (GDM), pericardial effusion, heart failure, limb edema, serum creatinine, white blood cell count, low platelet counts within one week before delivery, SBP on the first postpartum day, and postpartum antihypertensive use. Additionally, one risk factor from the index pregnancy was included, which was antihypertensive use before 20 weeks. The model demonstrated better discrimination, calibration, and a net benefit across a wide range of recurrent preeclampsia risk thresholds. Furthermore, the model has been translated into a clinical risk calculator, enabling clinicians to calculate individualized risks of recurrent preeclampsia. Conclusion: Our study demonstrates that a predictive tool utilizing routine clinical and laboratory factors can accurately estimate the risk of recurrent preeclampsia. This predictive model has the potential to facilitate shared decision-making by providing personalized and risk-stratified care.

Objective: Compare the clinical severity of second preeclampsia with the first preeclampsia. Methods: This retrospective longitudinal cohort study was conducted in three teaching hospitals in Guangzhou, where there were a total of 296 405 deliveries between 2010 and 2021. Two consecutive singleton deliveries complicated with preeclampsia were included. Clinical features, laboratory results within 1 week before delivery, and maternal and neonatal outcomes of both deliveries were collected. Univariate analyses were made using paired Wilcoxon tests and McNemar tests. Multivariable logistic regression and generalized linear models were performed to assess the association of adverse maternal and neonatal outcomes with second preeclampsia. Results: A total of 151 women were included in the study. The mean maternal age was 28 and 33 years for the first and second deliveries, respectively. The proportion of preventive acetylsalicylic acid use was 4.6% for the first delivery and 15.2% for the second delivery. No significant differences were observed in terms of blood pressure on admission, gestational weeks of admission and delivery, application of perinatal antihypertensive agents, rates of preterm delivery, and severe features between the two occurrences. However, the rates of heart disease, edema, and admission to the ICU were lower, and hospital stays were shorter in the second preeclampsia compared with the first preeclampsia. Sensitivity analysis conducted among women who did not use preventive acetylsalicylic acid yielded similar results. After adjusting for potential confounding variables, the occurrence of second preeclampsia was associated with significantly decreased risks of heart disease, edema, complications, and admission to the NICU, with odds ratios ranging between 0.157 and 0.336. Conclusion: Contrary to expectations, the second preeclampsia did not exhibit worse manifestations or outcomes to the first occurrence. In fact, some clinical features and outcomes appeared to be better in the second preeclampsia.

Objectives: In clinical practice, the majority of alpha-thalassaemia cases arise from deletions of the alpha-globin genes. However, a subset of cases is attributed to rare haemoglobin variants, which can manifest with borderline or normal screening results, potentially leading to missed diagnoses in clinical practice. Methods: Blood samples were collected from family members and underwent haematological, DNA and RNA analysis. Results: The five-month-old proband presented a haematological phenotype consistent with Hb H disease. The mother's haematology profile was consistent with an alpha-thalassaemia carrier, while the father exhibited a borderline reduction in MCV and MCH. MALDI-TOF identified an abnormal alpha-chain in the proband. DNA analysis revealed a novel alpha-globin variant (HBA2:c.175C>A, alpha 58His>Asn, Hb DG-Nancheng) affecting the distal histidine in the family. The father and the mother had alpha-genotype of --(SEA)/alpha alpha and alpha(DG-Nancheng)alpha/alpha alpha, respectively; while the proband inherited both mutant alleles (--(SEA)/alpha(DG-Nancheng)alpha). Sequencing of cDNA from HBA2 gene identified an equal ratio of normal and mutant alleles. Conclusion: This rare case highlighted the importance of identifying rare haemoglobin variant during prenatal screening. The clinical and genetic data provides useful information on the pathogenicity of this variant and further insight into the role of distal histidine residue of alpha-globin.

OBJECTIVE:To carry out carrier screening for Spinal muscular atrophy (SMA) in reproductive-aged individuals from Dongguan region and determine the carrier frequency of SMN1 gene mutations.METHODS:Reproductive-aged individuals who underwent SMN1 genetic screening at the Dongguan Maternal and Child Health Care Hospital from March 2020 to August 2022 were selected as the study subjects. Deletions of exon 7 and 8 (E7/E8) of the SMN1 gene were detected by real-time fluorescence quantitative PCR (qPCR), and prenatal diagnosis was provided for carrier couples by multiple ligation-dependent probe amplification (MLPA).RESULTS:Among the 35 145 subjects, 635 were found to be carriers of SMN1 E7 deletion (586 with heterozygous E7/E8 deletion, 2 with heterozygous E7 deletion and homozygous E8 deletion, and 47 with sole heterozygous E7 deletion). The carrier frequency was 1.81% (635/35 145), with 1.59% (29/1 821) in males and 1.82% (606/33 324) in females. There was no significant difference between the two genders (χ² = 0.497, P = 0.481). A 29-year-old woman was found to harbor homozygous deletion of SMN1 E7/E8, and was verified to have a SMN1∶SMN2 ratio of [0∶4], none of her three family members with a [0∶4] genotype had clinical symptoms. Eleven carrier couples had accepted prenatal diagnosis, and one fetus was found to have a [0∶4] genotype, and the pregnancy was terminated.CONCLUSION:This study has determined the SMA carrier frequency in Dongguan region for the first time and provided prenatal diagnosis for carrier couples. The data can provide a reference for genetic counseling and prenatal diagnosis, which has important clinical implications for the prevention and control of birth defects associated with SMA.

BACKGROUND:PCR, Sanger sequencing and NGS are often employed for carrier screening of thalassemia but all of these methods have limitations. In this study, we evaluated a new third-generation sequencing-based approach termed comprehensive analysis of thalassemia alleles (CATSA) to explore the prevalence of thalassemia in the Dongguan region of southern China. METHODS:19,932 subjects were recruited for thalassemia screening and hemoglobin testing was performed for each of them. Routine PCR was performed for all the hemoglobin testing-positive subjects and CATSA was conducted for randomly selected subjects from hemoglobin testing-positive and negative subjects. RESULTS:In the 2716 subjects tested both by PCR and CATSA, 2569 had the same results and 147 had discordant results between the two methods. Sanger sequencing, specially designed PCR and MLPA confirmed the results of CATSA were all correct. In total, CATSA correctly detected 787 subjects with variants while routine PCR correctly detected 640 subjects with variants. CATSA yielded a 5.42% (147 of 2716) increment compared with routine PCR. In the 447 hemoglobin testing-negative subjects, CATSA identified pathogenic variants in 12 subjects. Moreover, CATSA identified a novel deletion (chr16:171262-202032) in the α-globin gene cluster. As a result, the deduced carrier frequency of α-thalassemia,β-thalassemia and α-/β-thalassemia was 5.62%, 3.85% and 0.93%, respectively. CONCLUSIONS:Our study demonstrated CATSA was a more comprehensive and precise approach than the routine PCR in a large scale of samples, which is highly beneficial for carrier screening of thalassemia. It provided a broader molecular spectrum of hemoglobinopathies and a better basis for a control program in Dongguan region.

INTRODUCTION:Though an increase in Hb A2 is one of the most key markers of β-thal carriers, a few independent cases are reported to show elevated Hb A2 levels caused by mutations in other genes beyond β-globin gene.METHODS:We reviewed the haematological indices of 47336 individuals to analyse the phenotype-genotype correlation and identified 1439 individuals (3.04%) positive in the elevation of Hb A2 . Globin and KLF1 genes analysis was performed, and further whole-exome sequencing was carried to dissect the genetic causes of those positive samples without β-thalassemic or KLF1 mutations.RESULTS:Of these 1439 individuals with elevated Hb A2 , 1381 had a molecular defect in globin genes, and most were β-thalassemic mutation; 10 had a molecular defect in KLF1 gene. Finally, among the 38 individuals without β-thalassemic or KLF1 mutations, 7 were identified to carried a loss-of-function mutation in SUPT5H.CONCLUSION:This study has provided a mutation spectrum of SUPT5H in a cohort screening leading to the elevation of Hb A2 . According to the previous observations that individuals with a combination of β-thal mutation and a SUPT5H variant might present moderate β-thaelassemia, these findings emphasized the importance of comprehensive molecular diagnosis to prevent birth defects of β-thaelassemia caused by rare mutations from modifier genes.

IntroductionMonosomy of terminal 16p13.3 is a relatively common subtelomeric abnormality, most affected individuals presented α-thalassemia, some also have mental retardation, developmental abnormalities and/or speech delay and facial dysmorphism, which is termed ATR-16 syndrome. Here, we reported two novel 16p13.3 deletions involving the α-globin gene cluster and multispecies conserved sequences (MCSs), causing only a phenotype of α-thalassemia.MethodsSamples were collected from members of the two families and were subjected to haematological and comprehensive genetic analysis.ResultsThe novel 108 Kb deletion in family A extends from the non-protein coding RNA gene (WASIR2) to the NPRL3 gene, removing MCS-R1 to R3. This deletion should arise de novo because it wasn’t detected in both parents. The novel 336 Kb deletion in family B should extend from telomere to ∼ chr16:336000, removing the entire α-globin gene cluster. Carriers of these two deletions presented with microcytosis and hypochromic red cells, in accordance with a phenotype of α0-thalassemia carrier.ConclusionOur study increases the mutation spectrum of α-thalassemia. MCSs deletion should be considered in clinical practice of thalassemia screening and diagnosis.

Abstract Objective: To develop and validate a nomogram to predict severe postpartum haemorrhage (PPH) following caesarean delivery. Design: Two-centre retrospective cohort study. Setting: Third Affiliated Hospital of Guangzhou Medical University and the Dongguan Maternal and Child Health Care Hospital. Population: Cesarean delivery patients from the Third Affiliated Hospital of Guangzhou Medical University were divided into a development cohort (n=11,037) and an internal validation cohort (n=4,673). Cesarean delivery patients from the Dongguan Maternal and Child Health Care Hospital (n=13,775) were enrolled in the external validation cohort. Methods: The nomogram was based on independent risk factors for severe PPH obtained by multivariate logistic regression. We evaluated the discrimination and calibration of the nomogram in the development and validation cohorts. The area under the receiver operating characteristic curve, the Hosmer-Lemeshow test, and calibration plots were used to assess the nomogram. Main outcome measures: Postpartum haemorrhage. Results: Severe PPH occurred in 4.5%, 4.3%, and 2.0% of the patients in the development, internal validation, and external validation cohorts, respectively. The nomogram used data including previous caesarean delivery, low pre-pregnancy weight, placenta previa, and placenta accreta spectrum disorders. The area under the curves of the nomogram in the internal and external validation cohorts were 0.915 (95% confidence interval, 0.893–0.938) and 0.799 (95% confidence interval, 0.764–0.834), respectively. Consistency between the predicted and actual probabilities was observed in both validation cohorts. Conclusions: The nomogram displayed good calibration and discrimination; thus, it can be used for screening and timely intervention by clinicians, thereby reducing severe PPH incidence.

Abstract Introduction Patients with a homozygous β0‐thalassemia mutation usually have a transfusion‐dependent β‐thalassemia major phenotype. However, some β‐thalassemia patients present with a relatively mild and even normal phenotype and always have a high level of Hb F induced by genetic modifiers. Methods In this study, we identified a homozygous β0‐thalassemia mutation (HBB: c.126_129delCTTT) in a 36‐year‐old pregnant woman. She had not presented any clinical symptoms of β‐thalassemia since birth. To investigate her unexpected mild phenotype, known genetic modifiers that ameliorate the severity of β‐thalassemia were analysed. Besides, we described the haematological changes during pregnancy. Results Two genetic modifiers (a heterozygous KLF1: c.519_525dup mutation; and two homozygous HBS1L‐MYB locus SNP variants: rs7776054 and rs9399137) were identified. However, she showed a gradually decreased level of Hb during pregnancy, and serious transfusion complication of hyperhaemolysis was induced and complicated the pregnancy. Conclusion This report is in accordance with previous findings that genetic modifiers can ameliorate the clinical severity of β‐thalassemia, even without obvious clinical symptoms in a prolonged steady state. However, the steady state can be disrupted during pregnancy. In addition, raising awareness of hyperhaemolysis among clinicians treating patients with thalassemia is necessary.

Objective To evaluate the incidence and types of chromosomal abnormalities in pregnancy loss and aborted fetuses with anomaly and compare the performance of subtelomeric MLPA and chromosomal microarray analysis (CMA) in these specimens. Methods Samples were collected from spontaneous miscarriages, stillbirths and aborted fetuses with anomaly between January 2015 and April 2019. Chromosomal abnormalities were detected using subtelomeric MLPA and CMA. Results Among the 172 miscarriage samples, CMA detected pathogenic chromosomal abnormalities in 88 cases. MLPA could identified all aneuploidies and most pathogenic CNVs, missing all polyploidies; Of the 30 stillbirths, one pathogenic CNV and two VOUS were identified by CMA, all of which were missed from MLPA; Of the 135 aborted fetuses with anomaly, CMA identified pathogenic chromosomal abnormalities in 32 fetuses (23.7%); 18.95% in fetuses with isolated, and 35% in fetuses with multiple anomalies. MLPA can identify all aneuploidies but missing most pathogenic CNVs. Conclusion Our systematical comparison of subtelomeric MLPA and CMA for chromosomal analysis of tissue from pregnancy loss and aborted fetuses with anomaly is useful for assessing clinical utility of these techniques. MLPA screening, coupled with CMA analysis, is a cost-effective approach to detect chromosomal abnormalities in miscarriage and anomalous fetuses. However, MLPA might not be appropriate for chromosome analysis in stillbirth without structural anomaly; further research with more samples is needed.

Background: Abnormal concentrations of maternal thyroid hormones are risk factors for some obstetrical complications. However, the influence induced by different types of maternal thyroid dysfunction on obstetrical complications and outcomes is still controversial. Methods: A total of 17219 pregnant women were drawn for a thyroid function test, including TSH and fT4. All participants were divided into 7 groups, on the basis of their blood tested results, and their pregnancy outcomes were followed up. The isolated hypothyroxinemia group was further divided into 2 cohorts, according to whether they receive levothyroxine. Pregnant complications and outcomes in two cohorts were observed and analyzed. Results: A total of 2621 (15.22%)were identified to have abnormal thyroid function, including 1150 with subclinical hypothyroidism, 526 with gestational transient thyrotoxicosis (GTT), 419 with subclinical hyperthyroidism, 336 with isolated hypothyroxinemia, 78 with hyperthyroidism and 76 with hypothyroidism. Compare to control group, subclinical hypothyroidism, subclinical hyperthyroidism, isolated hypothyroxinemia and hypothyroidism groups presented higher incidence in one or more complications of pregnancy, while, GTT and drug-controlled hyperthyroidism had little significant effect on pregnancy complications. In isolated hypothyroxinemia group, there were no significant difference outcomes between cohorts using levothyroxine and not treatment. Conclusions: Our results showed a high incidence rate of thyroid dysfunction in pregnant women, and subclinical hypothyroidism is most common, followed by GTT. In general, pregnant women with thyroid dysfunction presented high risk of pregnancy complications. Isolated hypothyroxinemia in pregnant women is a matter of concern and treatment with levothyroxine couldn’t improve pregnancy outcomes and obstetrical complications.

Objectives: To present the prenatal findings and the molecular cytogenetic analyses of a de novo interstitial deletion of 1q23.3 encompassing PBX1 gene. Case report: A 32-year-old woman (gravida 1, para 0) underwent amniocentesis at 26 weeks' gestation because of constant small fetal kidneys on prenatal ultrasound. Chromosome microarray analysis (CMA) detected a de novo deletion of 1.871 Mb at 1q23.3. The deletion encompassed 2 genes of PBX1 and LMX1A. PBX1 haploinsufficiency had been reported to lead syndromic congenital anomalies of kidney and urinary tract (CAKUT) in humans. Furthermore, at 31 weeks' gestation, borderline oligohydramnios and restricted fetal dimensions were revealed. Ultimately, the pregnancy was terminated at 32 weeks with a 1500-g female fetus presenting polydactyl of left hand. Conclusions: The shared phenotypes between this case and the previously published prenatal cases demonstrate that loss of function mutation in PBX1 should be suspicious in fetus with bilateral renal hypoplasia, oligohydramnios and intrauterine growth retardation (IUGR). (c) 2019 Taiwan Association of Obstetrics & Gynecology. Publishing services by Elsevier B.V.

In practice, gap-polymerase chain reaction (gap-PCR) and reversed dot-blot are the two most frequently used molecular diagnostic methods for α-thalassemia (α-thal) genotyping. Here, we describe three Chinese individuals from three unrelated families in whom a polymorphism on the α-globin gene cluster led to diagnostic pitfalls. During general molecular diagnosis of thalassemia, three individuals with unexplained results were found. Blood or chorionic villus samples were collected from these three individuals and their family members. Hematological investigations and genetic tests were performed. In Family 1, a polymorphism of HBA2: c.301-24delinsCTCGGCC at the annealing site of the forward primer used in the PCR-reverse dot-blot assay was identified, leading to allele drop-out during the PCR amplification process. In Family 2, a synonymous mutation of C>T substitution at codon 125 of the α2 gene (HBA2: c.376C>T) was identified, leading to the failure of PCR-reversed dot-blot for the HBA2: c.377T>C (Hb Quong Sze or Hb QS) mutation. In Family 3, the size of the PCR fragment from the α2-globin allele carrying the HBA2: c.-771_-428del mutation was smaller and nearly equal to the size of the fragment corresponding to the -α4.2 (leftward) deletion; we also found that the HBA2: c.-771_-428del mutation was linked to a known HBA1: c.-673A>G mutation in this family. In conclusion, diagnostic errors may be caused by technical pitfalls or inherent properties of the DNA sample. All logical steps should be taken to monitor and thus preclude such events.

The alpha(+)-thal deletion of 3.557kb (NG_000006.1: g.32745_36301del, -alpha(MAL3.5)), involving the entire alpha 2-globin gene, was identified in a Chinese family by multiplex ligation-dependent probe amplification (MLPA) followed by gap-polymerase chain reaction (gap-PCR) and sequencing. The proband, a compound heterozygote for this mutant gene and the Southeast Asian (--(SEA); NG_000006.1: g.26264_45564del19301) deletion, had a phenotype of Hb H disease [hemoglobin (Hb) 7.6g/dL, mean corpuscular volume (MCV) 60.0 fL, Hb H (beta 4) 0.7%, Hb Bart's (gamma 4) 2.4% and Hb A(2) 1.1%]; one of her sisters with same genotype showed a similar phenotype. Another two family members, who were carriers of this mutant gene, had a hematological phenotype of a silent alpha-thal. The 5' and 3' breakpoints of this deletion are located at the Y2 and Y1 boxes, respectively, therefore, it probably originated from an unequal crossover between these two homologous boxes. This mutation constitutes an additional heterogeneous defect causing alpha-thal in the Chinese population and would be valuable for elucidating the arrangement in the human alpha-globin gene cluster.

目的 建立一种快速检测SMN1基因纯合缺失的荧光探针熔解曲线分析方法.方法 收集正常成人的外周血标本94份,经MLPA检测过的已知SMN1和SMN2基因拷贝数的样本238例.采用荧光探针熔解曲线分析技术,分别对SMN1基因c.840C＞T和c.835-45G＞A进行基因分型,根据基因分型结果判断是否存在SMN1基因exon7纯合缺失.对SMN1基因和SMN2基因拷贝数比值为2∶2、2∶0和0∶2基因型样本进行10倍梯度稀释,评估检测体系的敏感性.选取SMN1基因和SMN2基因拷贝数比值为2∶2、2∶0和0∶2基因型样本各8例,不同时间检测3次,用于评估检测体系的重复性.结果 该方法可以方便地检测SMN1基因exon7纯合缺失,灵敏度至少为0.05 ng基因组DNA.应用此方法检测94例正常成人样本,未发现SMN1基因exon7纯合缺失;对238例已知基因型样本进行检测,结果显示准确率为100％.结论 荧光探针熔解曲线分析方法可以快速准确检测SMN1基因exon7纯合缺失,操作简单实用,适合大规模人群筛查和产前分子诊断.

We identified two novel δ-globin gene mutations in two families during routine thalassemia screening. One missense mutation at codon 73 on the δ-globin gene [δ73(E17)Asp→Val, HBD: c.221A>T] which results in a Hb A2 variant homologous to the β-globin gene variant called Hb Mobile [β73(E17)Asp→Val, HBB: c.221A>T] and we have named this variant Hb A2-Henan. The other is a nonsense mutation [δ7(A4)Glu→Stop, HBD: c.22G>T] which gives rise to a stop codon (TAG) at codon 7, resulting in δ0-thalassemia (δ0-thal). The Hb A2 in one individual with homozygous HBD: c.22G>T was absent.