The γ- to β-globin switch is intricately regulated during human ontogeny, and this process is manipulated for therapeutic approaches to treat β-hemoglobinopathies by activating γ-globin expression. Several genetic strategies to reactivate HbF have partially reversed the γ- to β-globin switch and ameliorated the clinical symptoms of β-hemoglobinopathies. However, whether the γ- to β-globin switch can be completely reversed remains unknown. Completely reversing the γ- to β-globin switch requires a thorough redirection of the locus control region (LCR) from interacting with the β-globin gene (HBB) to interacting with the γ-globin gene (HBG). Here, we found that disrupting the KLF1-mediated HBB-LCR interaction by mutating the CACCC motif in HBB leads to the release of the LCR and its retargeting to other β-like globin genes. Moreover, simultaneously disrupting the KLF1-mediated HBB-LCR interaction and the epigenetic repression of HBG by combined editing of the CACCC motif in HBB and the TGACCA motif in HBG reinforces the HBG-LCR interaction, resulting in almost exclusive γ-globin expression while nearly absent β-globin expression, achieving near complete reversal of the γ- to β-globin switch. This finding demonstrates the comprehensive regulation of the γ- to β-globin switch by gene competition and gene silencing mechanisms. This finding also suggests that silenced genes can be fully activated through the redirection of enhancer-promoter contacts and that the specificity of enhancer-promoter contact within chromosomal domains is achieved through the transcription factor clusters binding to enhancers and promoters. Combined editing of the CACCC&TGACCA motifs also offer a more optimal therapeutic strategy for β-hemoglobinopathies. ### Competing Interest Statement Wenji Dong is the chairman of Genmedicn Biopharma Ltd. The remaining authors declare no competing interests. Haihe Laboratory of Cell Ecosystem Innovation Fund, 22HHXBSS00008 CAMS Innovation Fund for Medical Sciences, 2021-I2M-1-016 State Key Laboratory of Common Mechanism Research of Major Diseases Platform and State Key Laboratory Special Fund, 2060204

Objective To analyze the mutation types of non-deletional hemoglobin H (Hb H) disease, explore the correlation between distinct genotypes and clinical phenotypes, and identify rare α-globin gene mutations, so as to provide evidence for clinical diagnosis and treatment, genetic counseling, and prenatal diagnosis. Methods Routine blood tests [hemoglobin (Hb), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC)] and Hb H analysis were performed on patients diagnosed and treated for α-thalassemia at the First Affiliated Hospital of Guangxi Medical University from January 2024 to January 2026. Gap-polymerase chain reaction (gap-PCR), fluorescence-based PCR melting curve assay (FCMA) and DNA sequencing were used for genetic analysis of thalassemia. Results Among 217 patients with non-deletional Hb H disease, 187 patients were identified as Hb H-CS (--SEA/αCSα) and 27 patients as Hb H-QS(--SEA/αQSα), and 3 patients carried rare gene mutations causing Hb H disease, including one case each of --SEA/αATG＞GTGα, --SEA/αCD90-92(-AGCTTCGG)α and --SEA/αCD30(-GAG)α. None of the patients were complicated with β-thalassemia. The results of routine blood tests showed that mild, moderate and severe anemia in the Hb H-CS group accounted for 22.99%, 64.71% and 12.30%, respectively; the corresponding proportions of mild, moderate and severe anemia in the Hb H-QS group accounted for 44.45%, 51.85% and 3.70%, respectively. The Hb levels were 107.30 g/L for the --SEA/αATG＞GTGα genotype, 88.40 g/L for the --SEA/αCD90-92(-AGCTTCGG)α genotype, and 73.70 g/L for the --SEA/αCD30(-GAG)α genotype, all accompanied by decreased MCV and MCH. Hb analysis revealed that the Hb H level was 13.60% (10.45%–15.90%) in the Hb H-CS group and 23.20% (17.30%–25.00%) in the Hb H-QS group, with a statistically significant difference between the two groups (P＜0.05). The Hb H levels of the --SEA/αATG＞GTGα, --SEA/αCD90-92(-AGCTTCGG)α and --SEA/αCD30(-GAG)α genotypes were 25.30%, 24.40% and 20.40%, respectively. Conclusion The predominant genotype of non-deletional Hb H disease is --SEA/αCSα, followed by --SEA/αQSα. Moderate anemia is the main clinical manifestation of non-deletional Hb H disease. The Hb H level in the Hb H-QS group is higher than that in the Hb H-CS group. Three cases of Hb H disease with genotypes of --SEA/αATG＞GTGα, --SEA/αCD90-92(-AGCTTCGG)α and --SEA/αCD30(-GAG)α are identified, presenting with mild to moderate anemia.

Objective To investigate the hematological parameters and clinical phenotypes of a heterozygote carrying the rare IVSⅠ-2 (T＞A) mutation in the β-globin gene. Methods Patients suspected of having β-thalassemia who underwent initial screening via routine blood tests [red blood cell count (RBC), hemoglobin (Hb), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), hematocrit (HCT), red blood cell distribution width (RDW)], and analyses of hemoglobin A2 (Hb A2) and fetal hemoglobin (Hb F) were enrolled in this study. Common β-thalassemia gene mutations were detected using fluorescent polymerase chain reaction (PCR) melting curve analysis. For individuals in whom no mutations were identified by the above methods, DNA sequencing was further performed to detect rare or unknown β-thalassemia gene mutations. Results Among 235 patients with β-thalassemia, 1 patient with rare β-thalassemia was identified. The patient harbored a heterozygous IVSⅠ-2 (T＞A) (HBB:c.92+2 T＞A) mutation in the β-globin gene coexisting with α-thalassemia of the αα/-α3.7 deletion type. The patient's routine blood test results were as follows: RBC 5.89×1012/L, Hb 107.00 g/L, MCV 55.40 fL, MCH 18.15 pg, MCHC 327.70 g/L, HCT 0.327 and RDW 0.18. The hemoglobin analysis results of the patient revealed an Hb A2 level of 5.0% and an Hb F level of 4.0%. Conclusion β-Thalassemia caused by the rare heterozygous IVS Ⅰ-2 (T＞A) mutation in the β-globin gene is reported for the first time in China. Clinically, the patient presents with mild anemia, decreased MCV and MCH levels, and elevated Hb A2. This mutation is relatively rare and prone to missed diagnosis.

Objective:To study the influence of KLF1 gene mutations on fetal hemoglobin(Hb F)levels and their effects on thalassemia.Methods:Cases undergoing thalassemia screening at the First Affiliated Hospital of Guangxi Medical University from March 2023 to December 2024 were selected.Hb analysis and blood routine analysis were performed using a Hb analyzer and a blood cell analyzer.Sanger sequencing was used to detect KLF1 gene mutations and γ-globin gene promoter region mutations,while fluorescence PCR melting curve method and Gap-PCR were used to detect the genotypes of α-and β-thalassemia gene mutations.Results:Among 100 cases with elevated Hb F,25 cases were detected with KLF1 gene mutations.Hb analysis showed that the Hb F levels in the 25 cases ranged from 4.9%to 75.9%,and the Hb A2 levels ranged from 1.2%to 6.3%.Blood routine results showed that the Hb levels ranged from 63.4 g/L to 144.00 g/L,the red blood cell count(RBC)ranged from 1.90×1012/L to 6.48×1012/L,the mean corpuscular volume(MCV)ranged from 44.20 fL to 102.90 fL,and the mean corpuscular hemoglobin(MCH)ranged from 13.70 pg to 35.40 pg.Gene mutation analysis identified 5 types of KLF1 gene mutations,including c.325 C＞T heterozygote,c.519-525dup CGGCGCC heterozygote,c.304T＞C heterozygote and homozygote,c.1022G＞A heterozygote,and c.939G＞T heterozygote.Among the 25 cases,14 cases were compounded with β-thalassemia and 7 cases were compounded with α-thalassemia.Con-clusion:It is the first time in China that rare KLF1 gene mutations c.325 C＞T and c.939G＞T have been found to cause elevated Hb F levels,and they can alleviate the severity of anemia when combined with thalassemia.The KLF1 gene mutations c.519-525dup CGGCGCC,c.304T＞C,and c.1022G＞A have the same effects.

Naringin is a main flavonoid in citrus industry, and it is also a bitter matter in fruit juice. In our previous work, a mutant strain can produce alpha-L-rhamnosidase with good hydrolysis efficiency for hesperidin other than naringin. To extend its function based on our present study, we have explored the reason why the efficiency of catalysing hesperidin into hesperetin 7-O-glucoside than catalysing naringin into prunin. As a result, we found five key amino acid residues were responsible for the difference by molecular docking and molecular dynamics simulations. The terminal rhamnose of naringin is bound to the inside of the active pocket of the enzyme, and the addition of two new hydrogen bonds come out after the hydrolysis of naringin. So the active pocket began to contract, making it difficult for the terminal rhamnose to detach from it. 5 key amino acid residues, THR203, GLU158, LYS276, GLN133, and ILE258 were identified, and obtained five new mutants by mutating them into ALA. Among them, the mutant strain of K276A showed higher conversion efficiency from naringin to prunin along with higher thermal stability. This work not only improved the affinity of alpha-L-Rhamnosidase for catalysing bittering matter of naringin and enhances its potential application in the food and pharmaceutical industries, but also provided an effective technological strategy for improving the affinity of other enzymes.

Objective:To analyze the rare β-globin gene poly A(AATAAA＞AATAGA)mutation that causes β-thalassemia,and provide a basis for clinical diagnosis and treatment.Methods:Patients undergoing thalassemia screening and diagnosis at the First Affiliated Hospital of Guangxi Medical University from October 2023 to De-cember 2024 were collected.Blood routine and hemoglobin(Hb)analyses were performed on these patients.The fluorescence PCR melting curve assay(FCMA)was applied to detect common β-thalassemia gene mutations,while Sanger sequencing technique was used to detect unknown or rare β-thalassemia gene mutations.Results:A total of 282 cases of β-thalassemia heterozygotes were detected,including 2 cases with the rare β-globin gene poly A(AATAAA＞AATAGA,HBB:c.+112 A＞G)mutation.Their blood routine results showed that hemoglo-bin(Hb)was 130.00 g/L-135.00 g/L,mean corpuscular volume(MCV)was 61.90 fL-65.90 fL,mean corpuscular hemoglobin(MCH)was 19.20 pg-21.70 pg,mean corpuscular hemoglobin concentration(MCHC)was 311.00 g/L-329.00 g/L,and hematocrit(HCT)was 0.41 l/L-0.42 L/L.Notably,MCV and MCH levels were decreased.The results of hemoglobin(Hb)analysis showed that the level of Hb A2 was elevated,ranging from 4.6%to 4.9%.When this mutation combined with other β-thalassemia mutations,mild to moderate anemia was observed clini-cally.Conclusion:The rare β-globin gene poly A(AATAAA＞AATAGA)mutation was first identified in China.It causes β⁺-thalassemia,and needs to be attached importance to clinical diagnosis and treatment,genetic counsel-ing,and prenatal diagnosis.

BackgroundHemoglobin (Hb) is an important protein in red blood cells and a crucial diagnostic indicator of diseases, e.g., diabetes, thalassemia, and anemia. However, there is a rare report on methods for the simultaneous screening of diabetes, anemia, and thalassemia. Isoelectric focusing (IEF) is a common separative tool for the separation and analysis of Hb. However, the current analysis of IEF images is time-consuming and cannot be used for simultaneous screening. Therefore, an artificial intelligence (AI) of IEF image recognition is desirable for accurate, sensitive, and low-cost screening.ResultsHerein, we proposed a novel comprehensive method based on microstrip isoelectric focusing (mIEF) for detecting the relative content of Hb species. There was a good coincidence between the quantitation of Hb via a conventional automated hematology analyzer and the one via mIEF with R2 = 0.9898. Nevertheless, our results showed that the accuracy of disease diagnosis based on the quantification of Hb species alone is as low as 69.33%, especially for the simultaneous screening of multiple diseases of diabetes, anemia, alpha-thalassemia, and beta-thalassemia. Therefore, we introduced a ResNet1D-based diagnosis model for the improvement of screening accuracy of multiple diseases. The results showed that the proposed model could achieve a high accuracy of more than 90% and a good sensitivity of more than 96% for each disease, indicating the overwhelming advantage of the mIEF method combined with deep learning in contrast to the pure mIEF method.SignificanceOverall, the presented method of mIEF with deep learning enabled, for the first time, the absolute quantitative detection of Hb, relative quantitation of Hb species, and simultaneous screening of diabetes, anemia, alpha-thalassemia, and beta-thalassemia. The AI-based diagnosis assistant system combined with mIEF, we believe, will help doctors and specialists perform fast and precise disease screening in the future.

The isoelectric focusing has realized various improvements, including the protocols and creation of mIEF (microcolumn isoelectric focusing) instruments with excellent sensitivity for screening of diabetes and beta thalassemia. However, the problem of manual sample loading and hydration for the mIEF limits the operational capacity for stably detecting and quantitating most abnormal hemoglobin (Hb). Herein, we provided a high stable sample loading protocol for analysis of alpha thalassemia and Hb variants. In contrast to the previous volume of 20 mu l, a 100 mu l blood sample solution in this protocol was optimized with mixture of 6.4-7.5 and 3-10 pH carrier ampholytes, pI markers and loaded for 30 mins IPG microcolumn hydration. The hydrated microcolumn was then automatically loaded onto the mIEF chip array to which CH3COOH and NH4OH act as anodic and cathodic solutions. Lastly, the IEF was run for 9 mins. Hb H, Barts, A1c, F, A2 and CS were simultaneously separated and focused with higher resolution and sensitivity in quantifying H and Barts as low as 0.6 and 0.5 % respectively. Accordingly, there was an enhanced stability and linearity with a rapid assay time of 45 secs per sample. Moreover, analysis showed a fitting linear relationship with conventional technology at R2 = 0.9803 for H and R2 = 0.9728 for Barts thereby indicating greater accuracy confirmed by the AUC. Hence, the developed protocol could simply be employed for high stable and throughput batch sample loading of hydration, and accurate separation and quantitation of Hb variants for alpha and beta thalassemia.

Background The global prevalence of α-thalassemia necessitates effective newborn screening strategies due to its severe clinical consequences. Traditional methods such as liquid chromatography (LC), capillary electrophoresis (CE), and isoelectric focusing (IEF) face limitations, including low separation efficiency, poor sensitivity for detecting Hb Bart's, and time-intensive operations, particularly with dried blood spots (DBS). These limitations hinder timely and accurate screening. This study addresses the need for a more efficient, sensitive, and rapid method for detecting Hb Bart's in newborns. Results We enhanced IEF separation and sensitivity by designing a microfluidic IEF (mIEF) system with shortened columns and employing a coelution sample loading technique using oil-sealed blood spots for rapid sample pretreatment. Our experiments demonstrated significant improvements: the total analysis time was reduced from 1110 min (IPG IEF) and 46 min (LC) per batch to 36 min per batch. For individual samples, the focusing time decreased from 6 min (previous mIEF) to 3 min, with the microcolumn length shortened by 50 %, from 30 mm to 15 mm. The developed method showed excellent consistency with clinical Bart's detection and PCR diagnosis, achieving 100 % sensitivity and 98 % specificity for α-thalassemia screening. Significance and novelty Our novel mIEF method provides an efficient, sensitive, and rapid tool for screening newborns for α-thalassemia. This advancement addresses the limitations of traditional techniques, improving early diagnosis and intervention strategies and ultimately enhancing health outcomes for at-risk newborns.

Traditional capillary isoelectric focusing (cIEF), liquid chromatography (LC) and capillary zone electrophoresis (CZE) still suffered from low resolution for hemoglobinopathy screening. Herein, a 30-mm pH 5.2-7.8 micro-column IEF (mIEF) array chip was developed for hemoglobinopathy screening. As a proof of concept, adult beta-thalassemia was chosen as a model disease. In the method, blood samples were hemolyzed via hemolysin so-lution and loaded into the microcolumn. The experiments showed that (i) the species of Hb A, F, A2 and variants were clearly separated in the chip, and the resolution was greatly higher than the ones of LC/CZE/cIEF; (ii) up to 24 samples could be simultaneously analyzed in 12-min run; (iii) the intraday and interday RSDs were respec-tively 3.32-4.91 % and 4.07-5.33 %. The assays of mIEF to total 634 samples were compared with the ones of LC (n = 327) and PCR (n = 307). The cutoff of 3.5 % HbA2 led to the sensitivity of 100 % and specificity of 89.1 % for the mIEF-based screening; and there was 96.7 % coincidence between the methods of mIEF and PCR if refer Hb A2 and F. The method had the merits of facility, efficiency, specificity and sensitivity in contrast to the currently-used methods, implying its potential to screening of beta-thalassemia and hemoglobinopathies.

目的:探讨电神经肌肉刺激联合生物反馈对阴道口闭合不全患者性生活质量的影响.方法:选取我院2020年1月—12月收治的80例阴道口闭合不全患者为研究对象,将患者根据电脑盲选法分为两组.对照组患者行阴道内电神经肌肉刺激联合生物反馈治疗,研究组患者行阴道内外电神经肌肉刺激联合生物反馈治疗,对比两组患者的治疗总有效率及性生活质量改善情况.结果:研究组患者治疗总有效率(95.00％)较对照组(77.50％)更高(P<0.05);治疗前两组患者性生活各指标评分比较无统计学差异(P>0.05),治疗后研究组患者性生活各指标评分较对照组更高(P<0.05).结论:阴道内外电神经肌肉刺激联合生物反馈可有效促进阴道口闭合,提高患者的性生活质量,对促进阴道口闭合不全患者病情康复具有积极意义.

目的:研究β-珠蛋白基因CD114(CTG＞CCG,HBB:c.344T＞C)突变导致β-地中海贫血的血液学特点和临床特征.方法:对地中海贫血筛查疑为β-地中海贫血的病例,采用血细胞常规检测及高效液相色谱法(HPLC)血红蛋白电泳进行血液学表型分析;荧光PCR熔解曲线法、Gap-PCR法和DNA测序进行α-和β-地中海贫血基因分析.结果:共检出440例β-地中海贫血杂合子病例,其中检出1例罕见的β-珠蛋白基因突变杂合子病例,基因分析为β-珠蛋白基因CD114(CTG＞CCG,HBB:c.344T＞C)杂合子突变.该病例为女性,29岁,有中度贫血,血常规结果:血红蛋白(Hb)78.3 g/L,红细胞(RBC)3.65xl012/L,平均红细胞体积(MCV)66.52 fL,平均红细胞血红蛋白量(MCH)21.48pg,红细胞平均血红蛋白浓度(MCHC)322.9 g/L.其余439例β-地中海贫血杂合子病例表现为轻度贫血,血常规结果:Hb(115.2±12.46)g/L,RBC(5.79±0.68)x 1012/L,MCV(64.27±5.48)fL,MCH(20.10±1.94)pg,MCHC(312.58±8.41)g/L.不同性别的β-地中海贫血杂合子病例血常规分析结果比较,除MCHC外,其他指标差异均有统计学意义(P＜0.05).结论:首次在中国人群中发现罕见β-珠蛋白基因CD114(CTG＞CCG,HBB:c.344T＞C)突变导致β-地中海贫血.此突变类型罕见,临床上容易漏诊,提示在临床诊疗、遗传咨询和产前诊断过程中要重视这种类型的突变.

Guangxi Province is located in the southwest of the People's Republic of China (PRC). The province has a population of 50.12 million with a birth rate of 13.31%. Thalassemia is a major health problem in Guangxi Province. About 20.0-25.0% of the population carries thalassemia genes, which is acknowledged to be the highest prevalence in China. National and provincial programs for thalassemia prevention and control have been introduced. Premarital screening and prenatal diagnosis (PND) for the prevention of thalassemic fetuses are available. Blood transfusions, iron chelation therapy, and stem cell transplantation are also available for transfusion-dependent thalassemic patients.

目的 探讨经皮神经电刺激(TENS)联合加速康复外科(ERAS)多模式镇痛在腹腔镜全子宫切除术后的镇痛效果.方法 选取拟行腹腔镜全子宫切除术患者60例,随机分为ERAS组和ERAS+TENS组,每组30例.比较两组患者术后4、8、16、24 h疼痛程度视觉模拟评分(VAS)和Ramsay镇静评分.于患者麻醉诱导前、手术开始30 min、术后清醒时检测血清β-内啡肽水平.比较两组患者术中出血量、手术时间、术中麻醉药物用量、术后48 h恶心呕吐发生率、术后排气时间、术后住院时间.结果 与ERAS组相比,ERAS+TENS组术后16 h内疼痛VAS评分下降,术后8 h和16 h时Ramsay镇静评分较升高(P<0.05).手术开始30 min和术后清醒时,ERAS+TENS组β-内啡肽水平较ERAS组明显升高(P<0.05).两组患者术中出血量、手术时间、术后住院时间、术后排气时间、术中异丙酚和瑞芬太尼用量、恶心呕吐发生率比较差异均无统计学意义(P>0.05),但ERAS+TENS组非切口痛发生率明显下降(P<0.05).结论 TENS联合ERAS多模式镇痛可降低腹腔镜全子宫切除术患者术后的疼痛感,提高镇静舒适度,但无法减少阿片类药物用量.

目的:分析γ珠蛋白基因Aγ-225～-222AGCA缺失突变复合重型β-地中海贫血的临床特征.方法:收集2020年7月至2022年1月在广西医科大学第一附属医院门诊诊断为地中海贫血患者300例,采用血细胞分析仪进行血常规检测,高效液相色谱法(HPLC)测定胎儿血红蛋白(Hb F)和血红蛋白A2(Hb A2);DNA测序方法分析γ珠蛋白基因突变;采用荧光PCR熔解曲线法和Gap-PCR法检测地中海贫血基因突变.结果:在300例Hb F升高的病例中,共检出16例Aγ-225～-222缺失突变复合重型β-地中海贫血,其中7例轻度贫血,Hb为90～111.9g/L、MCV 65.9～80.31 fL、MCH 20.22～28.06 pg;中度贫血7例,Hb 60.7～87 g/L、MCV 62.9～86.24 fL、MCH 19.4～27.21 pg;重度贫血2例,其中 1 例Hb 47.3 g/L、MCV 77.31 fL、MCH 24.8 pg,另1 例 Hb 55.5 g/L、MCV 75.26 fL、MCH 24.49 pg.Hb 分析 16 例的 Hb F 为 8.6％～85.6％,Hb A2为 2.6％～9.3％.基因型分析显示,16例Aγ-225～-222AGCA缺失突变.16例β地中海贫血基因突变类型为βCD41-42/β-28 7例、βCD17/β-28 3例、β-28/β-282例、βIVS-I-1/β-281例、βCD71-72/β-28 1例、βCD41-42/βCD17 1例和βIVS-11-654/β-28 1例.结论:Aγ-225～-222AGCA缺失突变复合重型β-地中海贫血的病例大部分表现为轻度至中度贫血,该缺失突变可使重型β-地中海贫血患者的贫血程度得到改善.

目的:探讨香港型地中海贫血(HKαα)及其复合-α4.2缺失型α地中海贫血(HKαα/-α4.2)的临床表型和血液学特征,为临床诊疗和遗传咨询提供参考.方法:收集2021年5月至2022年3月在广西医科大学第一附属医院进行地中海贫血检测的病例,采用血细胞分析仪进行血液学分析,高效液相色谱法(HPLC)进行血红蛋白分析;采用跨越断裂点聚合酶链反应(Gap-PCR)和荧光PCR熔解曲线法(FCMA)进行α和β地中海贫血基因分析.结果:共检出165例患者,其中-α3.7或-α4.2缺失型α地中海贫血杂合子90例,--SEA/-α3.7或-SEA/-α4.2缺失型血红蛋白H(Hb H)病75例.在165例中检出1例罕见的HKαα/-α42及5例HKαα.基因型为HKαα/-α4.2的患者血常规检测结果:红细胞计数(RBC)5.59×1012/L,血红蛋白(Hb)166 g/L,红细胞平均容积(MCV)88.60 fL,红细胞平均血红蛋白量(MCH)29.70 pg,红细胞平均血红蛋白浓度(MCHC)335.00 g/L;5例HKαα的患者血常规结果:RBC(4.77±0.26)×1012/L,Hb(135.68±4.37)g/L,MCV(86.25±4.05)fL,MCH(28.47±1.12)pg,MCHC(330.48±3.75)g/L.6例患者中,Hb分析结果均未见异常,均无贫血表现,无黄疸及肝脾肿大.基因分析结果:6例患者αααanti4.2基因检测结果均为阳性,1例基因型为HKαα/-α4.2,5例基因型为HKαα.结论:首次发现HKαα/-α4.2患者无贫血症状,且血液学检测正常,提示此类病例在临床上较容易漏诊和误诊.

The involvement of circRNAs in β-thalassemia and their actions on fetal hemoglobin (HbF) is unclear. Here, the circRNAs in β-thalassemia carriers with high HbF levels were comprehensively analyzed and compared with those of healthy individuals. Differential expression of 2183 circRNAs was observed and their correlations with hematological parameters were investigated. Down-regulated hsa-circRNA-100466 had a strong negative correlation with HbF and HbA2. Bioinformatics was employed to construct a hsa-circRNA-100466‑associated competing endogenous RNA (ceRNA) network to identify hub genes and associated miRNAs. The hsa-circRNA-100466▁miR-19b-3p▁SOX6 pathway was identified using both present and previously published data. The ceRNA network was verified by qRT-PCR analysis of β-thalassemia samples, RNA immunoprecipitation of K562 cell lysates, and dual-luciferase reporter analysis. qRT-PCR confirmed that hsa-circRNA-100466 and SOX6 were significantly down-regulated, while miR-19b-3p was up-regulated. Hsa-circRNA-100466, miR-19b-3p, and SOX6 were co-immunoprecipitated by anti-argonaute antibodies, indicating involvement with HbF induction. A further dual-luciferase reporter assay verified that miR-19b-3p interacted directly with hsa-circRNA-100466 and SOX6. Furthermore, spearman correlation coefficients revealed their significant correlations with HbF. In conclusion, a novel hsa-circRNA-100466▁miR-19b-3p▁SOX6 pathway was identified, providing insight into HbF induction and suggesting targets β-thalassemia treatment.

目的:分析罕见基因突变导致血红蛋白H(Hb H)病的临床和血液学特征.方法:收集2020年5月至2021年2月到广西医科大学第一附属医院就诊、经Hb分析确诊为Hb H病的88例病例,应用跨越断裂点PCR(Gap-PCR)、荧光PCR熔解曲线法(FCMA)检测常见的缺失型和非缺失型α-地中海贫血基因突变,应用二代测序技术检测罕见基因突变.结果:在88例病例中发现3例由-α27.6缺失型突变导致Hb H病的病例,3例均为广西人.病例1为11个月大的女婴,病例2为44岁女性,病例3为10岁男童,3例均表现为中度小细胞低色素贫血,Hb平均值为76.8 g/L,红细胞平均容积(MCV)、红细胞平均血红蛋白量(MCH)、红细胞平均血红蛋白浓度(MCHC)均下降.无黄疸、肝脾肿大和输血史.Hb分析结果显示,3例病例有Hb Bart's和Hb H,依次为Hb Bart's 7％,Hb H 4％和Hb H 3％.基因检测结果均显示为α-珠蛋白基因27.6 kb缺失复合东南亚缺失型突变(--SEA/-α27.6).结论:在广西地区发现-α27.6缺失型突变,当复合东南亚缺失型突变可导致Hb H病,临床上表现为中度贫血,Hb分析有Hb Bart's和Hb H.此类型基因突变较为罕见,容易漏诊,需进一步的基因分析确诊.