Immunodeficiency, centromeric instability, and facial anomalies syndrome (ICF) is a rare genetic disease characterized by hypogammaglobulinemia, T cell immune deficiency with age, pericentromeric hypomethylation, facial abnormalities, and intellectual disability. This study aimed to investigate the phenotype and immune function of a girl with ICF2, identify her genetic defect, and explore the potential pathogenic mechanisms of the disease. We identified a homologous deletion mutation in this girl, which involves exons 1-5 and part of introns 1 and 6 of the ZBTB24 gene (NG_029388.1: g.2831_18,995del). This ZBTB24 variant produces a severely truncated ZBTB24 protein that lacks the BTB, A-T hook and eight zinc fingers. The above changes may lead to abnormal transcriptional function of the ZBTB24 protein. Karyotype analysis showed fragile sites and entire arm deletions were detected on chromosomes 1 and 16 and triradials on chromosome 16. The novel multi-exon deletion of ZBTB24 causes immunodeficiency, severe pneumonia and centromeric instability in the patient. During the follow-up, the patient’s pneumonia continued to progress despite receiving intravenous immunoglobulin (IVIG) replacement and anti-infective therapy. These results indicated that this novel multi-exon deletion variant of ZBTB24 may be the genetic etiology of ICF2. The discovery of this novel mutation expands the mutation spectrum of the ZBTB24 gene and improves our understanding of the molecular mechanisms underlying ICF.

Background: Mild to moderate thalassemia trait (TT) and iron deficiency anemia (IDA) are the most common conditions of microcytic hypochromic anemia (MHA) and they exhibit highly similar clinical and laboratory features. It is sometimes difficult to make a differential diagnosis between TT and IDA in clinical practice. Therefore, a simple, effective, and reliable index is needed to discriminate between TT and IDA. Methods: Data of 598 patients (320 for TT and 278 for IDA) were enrolled and randomly assigned to training set (278 of 598, 70%) and validation set (320 of 598, 30%). Stepwise discriminant analysis was used to define the best diagnostic formula for the discrimination between TT and IDA in training set. The accuracy and diagnostic performance of formula was tested and verified by receiver operating characteristic (ROC) analysis in validation set and its diagnostic performance was compared with other published indices. Results: A novel formula, Thalassemia and IDA Discrimination Index (TIDI) = –13.932 + 0.434 × RBC + 0.033 × Hb + 0.025 ×MCHC + 53.593 × RET%, was developed to discriminate TT from IDA. TIDI showed a high discrimination performance in ROC analysis, with the Area Under the Curve (AUC) = 0.936, Youden’ s index = 78.7%, sensitivity = 89.5%, specificity = 89.2%, respectively. Furthermore, the formula index also obtained a good classification performance in distinguishing 5 common genotypes of TT from IDA (AUC from 0.854–0.987). Conclusion: The new, simple algorithm can be used as an effective and robust tool for the differential diagnosis of mild to moderate TT and IDA in Guangxi region, China.

The red blood cell distribution width (RDW) to albumin ratio (RAR), a novel indicator of inflammation, is known to be associated with a poor prognosis in various diseases. The purpose of this study was to investigate whether RAR is also associated with mortality in critically ill patients with acute kidney injury (AKI). A retrospective observational study was conducted using the Medical Information Mart for Intensive Care III (MIMICIII) database, which contains comprehensive clinical data relating to patients with AKI between 2001 and 2012. Patients were grouped into quartiles (Q1-Q4) according to the RAR. All-cause mortality was then compared across the four groups using Kaplan-Meier analysis. Cox proportional hazard models and subgroup analyses were use to investigate RAR and the prognosis of patients with AKI. A total of 3826 critically ill patients with AKI were included in this study. Based on Kaplan-Meier curve analysis, the patient group with a high-RAR exhibited elevated rates of mortality at 28 days (log-rank p < 0.001). Multivariable Cox proportional hazard models identified RAR as a significant predictor of mortality at 28 days (Hazard ratio, HR (95% Confidence Interval, CI) 1.07 (1.03-1.11), p < 0.001), in the hospital (HR (95% CI) 1.08 (1.05-1.12), p < 0.001), and in the intensive care unit (ICU) (HR (95% CI) 1.06 (1.02-1.11), p = 0.004). Furthermore, the subgroup analysis showed that the effect of the RAR was significantly greater in patients with diabetes than in those without diabetes (p for interaction = 0.005). As the RAR increased, the mortality rate within 28 days of hospitalization and in the ICU also increased. Thus, the RAR has the potential to become an important and practical indicator for identifying a poor prognosis in critically ill patients with AKI.

BACKGROUND:Hemophilia A (HA) is an X-linked recessive genetic disorder caused by pathogenic variations of the factor VIII -encoding gene, F8 gene. Due to the large size and diverse types of variations in the F8 gene, causative mutations in F8 cannot be simultaneously detected in one step by traditional molecular analysis, and genetic molecular diagnosis and prenatal screening of HA still face significant difficulties and challenges in clinical practice. Therefore, we aimed to develop and validate an efficient, accurate, and time-saving method for the genetic detection of HA. METHODS:A comprehensive analysis of hemophilia A (CAHEA) method based on long-range PCR and long-read sequencing (LRS) was used to detect F8 gene mutations in 14 clinical HA samples. The LRS results were compared with those of the conventional methods to evaluate the accuracy and sensitivity of the proposed approach. RESULTS:The CAHEA method successfully identified 14 F8 variants in all probands, including 3 small insertion deletions, 4 single nucleotide variants, and 7 intron 22 inversions in a "one-step" manner, of which 2 small deletions have not been reported previously. Moreover, this method provided an opportunity to analyze the mechanism of rearrangement and the pathogenicity of F8 variants. The LRS results were validated and found to be in 100% agreement with those obtained using the conventional method. CONCLUSION:Our proposed LRS-based F8 gene detection method is an accurate and reproducible genetic screening and diagnostic method with significant clinical value. It provides efficient, comprehensive, and accurate genetic screening and diagnostic services for individuals at high risk of HA as well as for premarital and prenatal populations.

目的 探讨急性缺血性脑卒中(AIS)患者血清胶质纤维酸性蛋白(GFAP)和同型半胱氨酸(Hcy)水平与病情严重程度的相关性.方法 选取该院2022年1月至2023年2月收治的AIS患者110例作为观察组,另选取该院同期健康体检者59例作为对照组,比较观察组和对照组白细胞计数(WBC)、总胆固醇(TC)、甘油三酯(TG)、高密度脂蛋白胆固醇(HDL-C)、空腹血糖(FBG)、血清GFAP与Hcy水平;根据美国国立卫生院研究卒中量表(NIHSS)评分将110例AIS患者分为轻型组(73例)、中型组(32例)和重型组(5例),比较轻型组、中型组、重型组及不同住院时间患者的血清GFAP和 Hcy水平,采用Spearman相关分析GFAP、Hcy与AIS患者NIHSS评分、住院总时间及 WBC的相关性;构建受试者工作特征(ROC)曲线分析GFAP、Hcy单独及2项指标联合检测AIS的诊断价值.结果 观察组患者的TC水平、WBC高于对照组,HDL-C水平低于对照组,差异均有统计学意义(P<0.05);观察组血清GFAP、Hcy水平高于对照组(P<0.05);中型组患者GFAP、Hcy水平高于轻型组,重型组高于中型组和轻型组,差异均有统计学意义(P<0.05);住院时间在8～14 d患者的GFAP水平高于住院时间<8 d的患者,住院时间>14 d患者的GFAP水平高于住院时间<8 d的患者,差异均有统计学意义(P<0.05).Spearman相关分析结果显示,血清GFAP与Hcy呈正相关(r=0.253,P=0.001),GFAP水平与NIHSS评分、住院时间、WBC均呈现正相关(r=0.238、0.300、0.226,P<0.05),Hcy水平与NIHSS评分、WBC呈正相关(r=0.342、0.274,P<0.05).ROC曲线分析结果显示,GFAP、Hcy单独用于诊断AIS的 AUC分别为0.816、0.712,二者联合检测的 AUC为0.857.结论 AIS患者血清GFAP、Hcy水平均显著升高,且与AIS患者病情严重程度呈正相关,可用作临床AIS早期检测和评估病情严重程度的有效指标,具有一定临床参考价值.

Enlarged vestibular aqueduct is an autosomal genetic disease mainly caused by mutations in the SLC26A4 gene and includes non-syndromic and syndromic types. This study aimed to identify genetic defects in a Chinese patient with non-syndromic enlarged vestibular aqueduct (NSEVA) and to investigate the impact of variants on the severity of non-syndromic enlarged vestibular aqueduct. A male patient with NSEVA, aged approximately 6 years, was recruited for this study. The clinical characteristics and results of auxiliary examinations, including laboratory and imaging examinations, were collected, and 127 common hereditary deafness genes were detected by chip capture high-throughput sequencing. Protein structure predictions, the potential impact of mutations, and multiple sequence alignments were analyzed in silico. Compound heterozygote mutations c.1523_1528delinsAC (p.Thr508Asnfs*3) and c.422T>C (p.Phe141Ser) in the SLC26A4 gene were identified. The novel frameshift mutation c.1523_1528delinsAC produces a severely truncated pendrin protein, and c.422T>C has been suggested to be a disease-causing mutation. Therefore, this study demonstrates that the novel mutation c.1523_1528delinsAC in compound heterozygosity with c.422T>C in the SLC26A4 gene is likely to be the cause of NSEVA. Cochlear implants are the preferred treatment modality for patients with NSEVA and severe-to-profound sensorineural hearing loss Genetic counseling and prenatal diagnosis are essential for early diagnosis. These findings expand the mutational spectrum of SLC26A4 and improve our understanding of the molecular mechanisms underlying NSEVA.

Third-generation sequencing (TGS) has led to a brave new revolution in detecting genetic diseases over the last few years. TGS has been rapidly developed for genetic disease applications owing to its significant advantages such as long read length, rapid detection, and precise detection of complex and rare structural variants. This approach greatly improves the efficiency of disease diagnosis and complements the shortcomings of short-read sequencing. In this paper, we first briefly introduce the working mechanism of one of the most important representatives of TGS, single-molecule real-time (SMRT) sequencing by Pacific Bioscience (PacBio), followed by a review and comparison of the advantages and disadvantages of different sequencing technologies. Finally, we focused on the progress of SMRT sequencing applications in genetic disease detection. Future perspectives on the applications of TGS in other fields were also presented. With the continuous innovation of the SMRT technologies and the expansion of their fields of application, SMRT sequencing has broad clinical application prospects in genetic diseases detection, and is expected to become an important tool for the molecular diagnosis of other diseases.