Hemoglobin H (HbH) disease is a subtype of α-thalassemia typically caused by genetic defects in three out of four α-globin genes. To date, the genetic factors contributing to the highly heterogeneous clinical severity of HbH disease remain largely unknown. In this study, we perform targeted long-read sequencing (T-LRS) on a cohort of 591 HbH patients, aiming to profile the genomic variants and their haplotypes within the α/β-globin gene clusters and key erythroid genes. Phenotypic analysis confirms that non-deletional HbH patients generally exhibit more severe clinical manifestations compared to deletional ones. Moreover, we identify the co-inheritance of β-thalassemia mutations to be a mitigating factor for HbH patients, as reflected by higher hemoglobin levels and lower serum ferritin, suggesting the less imbalanced synthesis of α/β-globin among these patients. Furthermore, through haplotype phasing using long-sequencing reads, we find a haplotype of HS40 associated with milder clinical symptoms of HbH patients, validated by reporter gene assay, and that functional mutations in erythroid transcription factors BCL11A and MYB-HBS1L exert significant effects on β-thalassemia but not on HbH patients. This study presents the largest T-LRS study for α-thalassemia patients, which may provide insight into precise clinical diagnosis and phenotyping of HbH diseases.

Platelets act as a crucial indicator for monitoring hypercoagulability and thrombosis and a key target for pharmacological intervention. Genotype-phenotype association studies have confirmed that platelet traits are quantitatively regulated by multiple genes. However, there is currently a lack of genetic studies on the heterogeneity of platelet traits in β-thalassemia under a hypercoagulable state. Here, we studied the phenotypic heterogeneity of platelet count (PLT) and mean platelet volume (MPV) in a cohort of 1020 β-thalassemia patients. We further performed a functionally informed whole-genome sequencing (WGS) association analysis of common variants and rare variants for PLT and MPV in 916 patients through integrative analysis of WGS data and functional annotation data. Extreme phenotypic heterogeneity of platelet traits was observed in β-thalassemia patients. Additionally, the common variant-based gene-level analysis identified RNF144B as a novel gene associated with MPV. The rare variant analysis identified several novel associations in both coding and noncoding regions, including missense rare variants of PPP2R5C associated with PLT and missense rare variants of TSSK1B associated with MPV. In conclusion, this comprehensive and systematic whole-genome scan of platelet traits in the β-thalassemia cohort reveals the specific genetic regulation of platelet traits in the context of β-thalassemia, providing potential targets for intervention.

Background: Serum ferritin (SF) monitors secondary iron overload in beta-thalassemia (b-thalassemia). Transferrin (TRF) has been shown to reverse iron accumulation in experimental models, but its role in transfusion-dependent beta-thalassemia (TDT) patients remains unclear. This study aims to explore the relationship between TRF and SF in TDT patients and to reveal the unique connection between specific genotypes and iron metabolism, providing potential therapeutic targets for clinical practice. Methods: This cross-sectional study includes 817 TDT patients (b0/b0 genotype: n=560; b0/b+ genotype: n=257). We use genotype-phenotype analysis and employ logistic regression and restricted cubic spline (RCS) curves to assess the association between TRF and SF. Results: Significant differences were observed between the b0/b0 and b0/b+ genotypes in terms of age at first transfusion, transfusion requirements, chelation initiation age, reticulocyte count, red blood cell count, red cell distribution width-coefficient of variation (RDW-CV), fetal haemoglobin (HbF) level, splenomegaly, and SF. b0/b0 patients presented with more severe clinical phenotypes. SF was significantly associated with TRF, HbF, RDW-CV, and chelation therapy. RCS analysis revealed a dose-response relationship with a negative linear correlation between TRF and SF (OR=0.26, P<0.001), indicating that higher TRF levels are linked to lower SF risk. Conclusions: This study systematically confirms for the first time a significant negative correlation between high TRF levels and high SF risk in TDT patients. This new finding may help clinicians more effectively manage iron overload, especially in patients with different genotypes.

Despite the well-documented mutation spectra of β-thalassemia, the genetic variants and haplotypes of globin gene clusters modulating its clinical heterogeneity remain incompletely illustrated. Here, a targeted long-read sequencing (T-LRS) is demonstrated to capture 20 genes/loci in 1,020 β-thalassemia patients. This panel permits not only identification of thalassemia mutations at 100% of sensitivity and specificity, but also detection of rare structural variants (SVs) and single nucleotide variants (SNVs) in modifier genes/loci. The highly homologous regions of α-/β-globin gene clusters are then phased and 3 novel haplotypes in HBG1/HBG2 region are reported in this population of β-thalassemia patients. Furthermore, one of the haplotypes is associated with ameliorated symptoms of β-thalassemia. Similarly, 5 major haplotypes are identified in HBA1/HBA2 homologous region while one of them is found highly linked with deletional α-thalassemia mutations. Finally, rare mutations in erythroid transcription factors in DNMT1 and KLF1 associated with increased expression of fetal hemoglobin and reduced transfusion dependencies are identified. This study presents the largest T-LRS study for β-thalassemia patients to date, facilitating precise clinical diagnosis and haplotype phasing of globin gene clusters.

Objective:This study was aimed at investigating the carrier rate of, and molecular variation in, α- and β-globin gene mutations in Hunan Province.Methods:We recruited 25,946 individuals attending premarital screening from 42 districts and counties in all 14 cities of Hunan Province. Hematological screening was performed, and molecular parameters were assessed.Results:The overall carrier rate of thalassemia was 7.1%, including 4.83% for α-thalassemia, 2.15% for β-thalassemia, and 0.12% for both α- and β-thalassemia. The highest carrier rate of thalassemia was in Yongzhou (14.57%). The most abundant genotype of α-thalassemia and β-thalassemia was -α 3.7/αα (50.23%) and β IVS-II-654/β N (28.23%), respectively. Four α-globin mutations [CD108 (ACC>AAC), CAP +29 (G>C), Hb Agrinio and Hb Cervantes] and six β-globin mutations [CAP +8 (C>T), IVS-II-848 (C>T), -56 (G>C), beta nt-77 (G>C), codon 20/21 (-TGGA) and Hb Knossos] had not previously been identified in China. Furthermore, this study provides the first report of the carrier rates of abnormal hemoglobin variants and α-globin triplication in Hunan Province, which were 0.49% and 1.99%, respectively.Conclusion:Our study demonstrates the high complexity and diversity of thalassemia gene mutations in the Hunan population. The results should facilitate genetic counselling and the prevention of severe thalassemia in this region.

Objective:To analyze the costs and effectiveness of five common screening modes and genetic screening for thalassemia in China in order to find the optimal way and provide evidence for the implementation of thalassemia prevention and control projects in Hunan Province.Methods:From June 2020 to April 2021, 12 971 couples from 14 cities and autonomous prefectures in Hunan Province were selected as the study population. The diagnosis of thalassemia was based on the results of genetic testing. Results of routine blood test and hemoglobin electrophoresis were collected and analyzed. The efficacy of five screening modes, at the cut-off value of <80 fl or 82 fl for the mean corpuscular volume (MCV), was analyzed by positive predictive value, negative predictive value, Jorden index and cost-effectiveness ratio. Sensitivity analysis was used to assess the feasibility of genetic screening at different costs after fixing the costs of routine blood and hemoglobin electrophoresis. The five thalassemia screening models are as follows: Mode 1: The woman had a blood routine test first. If the result was positive, the spouse required a blood routine test. If both results were positive, a thalassemia gene test should be offered to the couple. Mode 2: Both husband and wife were screened by blood routine and hemoglobin electrophoresis. If one or both of them were positive, both would be tested for thalassemia gene. Mode 3: The couple received blood routine tests initially. If either was positive, both should receive hemoglobin electrophoresis testing. If either was positive, both parties will conduct thalassemia gene testing. Mode 4: The woman was screened by blood routine and hemoglobin electrophoresis. If any one of them was positive, the woman would be tested for thalassemia gene. If the gene test result was positive, the spouse should receive thalassemia gene. Mode 5: Both spouses conducted a blood routine test. If either was positive, both would conduct hemoglobin electrophoresis test. If both were positive, both spouses should receive thalassemia gene testing. Gene testing mode: The woman would be tested for thalassemia, and her spouse would have thalassemia test too if her result was positive.Results:When using MCV<80 fl as the cut-off for diagnosing thalassemia, the Youden indices of the five prenatal screening modes in Hunan Province were 0.551, 0.639, 0.898, 0.555 and 0.356, while when using MCV<82 fl as the cut-off, the Youden indices were 0.549, 0.629, 0.851, 0.548 and 0.356. When the MCV cut-off value was <80 fl, the missed diagnosis rates of the five screening modes were 44.44%, 0.00, 0.00, 18.52% and 62.96%, and the cost-effectiveness ratios were 21 709, 250 939, 76 870, 138 463 and 92 860 yuan (RMB)/couple, respectively. When the price of genetic testing was lower than 55 yuan (RMB), the cost-effectiveness ratio of genetic screening was lower than that of Mode 3.Conclusions:MCV<80 fl can be considered as the positive criteria in blood routine screening for thalassemia in Hunan Province, and the cost-effectiveness ratio of Mode 3 (the couple received blood routine tests initially. If either was positive, both should receive hemoglobin electrophoresis testing. If either was positive, both parties will conduct thalassemia gene testing) is the best. Genetic screening has certain advantages with the decreasing price.

Glucose-6-phosphate dehydrogenase (G6PD) deficiency is one of the most common X-linked enzymopathies caused by G6PD gene variant. We aimed to provide the characteristics of G6PD deficiency and G6PD gene variant distribution in a large Chinese newborn screening population. We investigated the prevalence of G6PD in China from 2013 to 2017. Then, we examined G6PD activity and G6PD gene in representative Chinese birth cohort to explore the distribution of G6PD gene variant in 2016. We then performed multicolor melting curve analysis to classify G6PD gene variants in 10,357 neonates with activity-confirmed G6PD deficiency, and DNA Sanger sequencing for G6PD coding exons if hot site variants were not found. The screened population, organizations, and provinces of G6PD deficiency were increased from 2013 to 2017 in China. The top five frequency of G6PD gene variants were c.1376G>T, c.1388G>A, c.95A>G, c.1024C>T, and c.871G>A and varied in different provinces, with regional and ethnic features, and four pathogenic variant sites (c.152C>T, c.290A>T, c.697G>C, and c.1285A>G) were first reported. G6PD deficiency mainly occurs in South China, and the frequency of G6PD gene variant varies in different regions and ethnicities.

Background: Glucose-6-phosphate dehydrogenase (G6PD) deficiency is one of the most common X-linked enzymopathies caused by G6PD gene mutation. We aimed to provide the characteristics of G6PD deficiency and G6PD gene mutation distribution in a large Chinese newborn screening (NBS) population.

OBJECTIVE:To summarize the molecular epidemiology of hemoglobinopathies in Yongzhou area of Hunan province in order to provide a basis for making the guidelines of local thalassemia prevention program.METHODS:Two thousand and two samples (1001 couples) were randomly recruited based on demographic data and distribution of ethnic groups. All samples were subjected to full blood count and analysis of hemoglobin and 6 common alpha-thalassemia mutations. Known beta-thalassemia mutations were screened in samples with beta-thalassemia trait. The remaining samples with positive phenotype and unknown mutations were subjected to DNA sequence analysis.RESULTS:Two hundred and forty individuals were found to be carriers of hemoglobinopathic mutations, which included 6 common alpha-thalassemia deletions, 9 common beta-thalassemia mutations and 3 common structural hemoglobin variants. One hundred and seventy-four mutant alleles for alpha-thalassemia were detected, which gave a carrier rate of 8.69%, of which 0.1% was due to HbH disease. Seventy mutant alleles for beta-thalassemia were detected, which gave a carrier rate of 3.50%. Seven subjects (0.35%) were identified as carriers of hemoglobin variants. The overall carrier rate for hemoglobinopathic mutations was 12.54% based on detection of 251 hemoglobinopathy mutant alleles. The overall carrier rate for alpha- and beta-thalassemia among ethnic Yaos was 25.00%, which was significantly higher than that of ethnic Han Chinese (11.14%, P< 0.01).CONCLUSION:The prevalence and mutation spectrum of hemoglobinopathies in Yongzhou area has been delineated for the first time.

Objective To explore an appropriate strategy for thalassemia screening by evaluating the correlations between the mean corpuscular volume (MCV),mean corpuscular hemoglobin (MCH),hemoglobin A2 (HbA2) and thalassemia among population of reproductive age,so as to improve the detection rate of thalassemia and decline its missed diagnosis and misdiagnosis rates.Methods We retrospectively analyzed 2,002 samples (including 234 cases of confirmed thalassemia and 1,768 cases of nonthalassemia) from the molecular epidemiological survey of hemoglobin-related diseases from April to May in 2016 among residents in Yongzhou.The sensitivity,specificity and accuracy of single index and combined indices of MCV,MCH and HbA2 in the screening and genetic diagnosis of thalassemia were statistically analyzed.Results The sensitivity of MCV,MCH and HbA2 in parallel testing (87.18％) was significantly higher than that of the three indexes in series testing (59.83％),with a statistically significant difference (P＜0.05).The specificity of MCV,MCH and HbA2 in series testing (99.55％) was obviously higher than that of the three index in parallel testing (87.16％),with a statistically significant difference (P＜0.05).The sensitivity of MCV and MCH in parallel testing (79.06％) was slightly lower than that of MCV,MCH and HbA2 in parallel testing (87.18％).The specificity (95.13％) and diagnostic coincidence rate (93.26％) of MCV and MCH were slightly lower than those of the three indexes in series testing (99.55％,94.91％),without statistically significant differences (all P＞0.05).Conclusions It is of great value to screening thalassemia through detection of MCV,MCH and HbA2.Parallel testing with MCV and MCH is the best way for thalassemia screening,with the advantages of simplicity,money-saving and efficiency.It is conducive to screening of thalassemia in grass-roots hosptials.

目的：比较氟康唑、伊曲康唑联合克霉唑治疗复发性外阴阴道假丝酵母菌病（RVVC）的疗效。方法收集2012年7月至2014年4月永州市各妇幼保健院妇科门诊的就诊患者，回顾性分析各种阴道炎所占比例。以就诊为RVVC的患者作为研究对象，氟康唑口服联合克霉唑阴道上药治疗的为对照组，伊曲康唑口服联合克霉唑阴道上药治疗的为试验组，其中对照组、试验组又各自分为2个亚组：对照1组、试验1组为卵泡期用药组，对照2组、试验2组为黄体期用药组。结果研究显示阴道感染人数7508例（33.32%），其中滴虫性阴道炎542例（7.22%）、外阴阴道假丝酵母菌病1254例（16.70%）、细菌性阴道病1509例（20.09%）、其他55.98%（包括混合感染、非特异性阴道炎等）。试验组总有效率为95.08%，对照组总有效率为69.35%,差异有统计学意义（P<0.05）。对照1组比对照2组治疗效果好（分别为59.09%、28.57%，P<0.05）；试验1组较试验2组疗效无明显差异（分别为75.86%、68.97%，P>0.05）。结论阴道炎患者中细菌性阴道病患者最多。伊曲康唑治疗RVVC有效率高。氟康唑的治疗效果与月经周期有关。

目的：比较超声经腹和经阴道诊断瘢痕妊娠的临床效果。方法选择2008年1月~2012年12月在我院收治的98例经病理确诊为瘢痕妊娠的患者，设高频彩超经阴道检查50例为研究组，彩超经腹部检查48例为对照组.比较两组检测的结果以及子宫组织及血流变化。并进行统计学分析。结果与对照组诊断准确率77.1%比较，研究组诊断准确率94.0%明显增高，差异显著，有统计学意义(P<0.05)。与对照组子宫组织及血流变化检出率比较(75.0%)，研究组子宫组织及血流变化检出率(100.0%)，差异显著，有统计学意义(P<0.05)。结论经阴道高频彩超诊断瘢痕妊娠符合率高，无创、经济、便捷。能为瘢痕妊娠的诊治提供科学的依据。是目前诊断瘢痕妊娠的金标准。

目的探讨高频超声多普勒对乳腺导管扩张症的诊断和鉴别诊断。方法回顾性分析37例经病理证实的乳腺导管扩张症的超声声像图表现。结果乳腺导管扩张症声像图表现大体分三型:低回声包块型、囊实性包块型、囊性导管扩展型,病灶多位于乳晕区及乳头深面,内部血流可见点状血流信号。结论高频超声多普勒检查对乳腺导管扩张症具有较高的诊断价值,但需与乳腺囊性增生病及乳腺癌等乳腺疾病鉴别。

目的 了解本地区女性各类病原体感染情况,为临床预防和治疗提供参考.方法 对8432例女性患者阴道分泌物做白带常规检查和革兰氏染色镜检.结果 8432例标本中,检查出检出滴虫353例,念珠菌1304例,线索细胞2030例,革兰氏阴性双球菌20例.门诊组4种病原体检出率各明显高于普查组(P均<0.01);清洁度Ⅰ口～Ⅱ口4种病原体检出率各明显低于清洁度Ⅲ口～Ⅳ口(P均<0.0I).结论 ①白带常规检查结合革兰氏染色镜检可提高各类病原体感染的诊断率;②清洁度正常时也要注意查找病原菌.

目的:分析2005年10月-2009年12月永州市新生儿先天性甲状腺功能减退症(CH)的筛查结果,总结筛查经验,指导今后的筛查工作.方法:采集新生儿足跟血,制成滤纸血片,采用时间分辨荧光分析法检测血片中TSH.初筛>20IU/ml为阳性组,TSH浓度在9-20IU/mL之间为疑似阳性,对疑似阳性进行样本复查,复查>9IU/mL以及初筛>20IU/mL的新生儿召回,抽静脉血确诊,采用电化学发光法检测FT3、FT4、TSH浓度.结果:101,028例血片标本中,初筛>20IU/mL及复查>9?IU/mL共计133例,确诊CH患儿70例,无漏筛病例;CH发病率为1/1443,明显高于国内平均发病率1/3009(u=11.28,P<0.01);初筛阳性组的检出率明显高于疑似阳性组的检出率,分别为1/2405及1/3608(u=5.29,P<0.01);复查后,疑似阳性组的阳性率比复查前显著降低,由1/83降低为1/153l(u=10.16,P<0.01).结论:1、永州为先天性甲状腺功能减退症的高发地区,应加强全市的新生儿筛查工作.2、对疑似阳性的样本进行血片复查可显著降低阳性召回率,并有效防止漏筛.3、检测滤纸干血片的TSH水平是CH筛查的有效措施,有助于CH患儿的早期诊治.

目的:了解永州市新生儿先天性甲状腺功能减退症(CH)及苯丙酮尿症(PKU)的发病情况.方法:采用酶联免疫法(ELISA)测定促甲状腺素(TSH)浓度,使用定量酶法测定苯丙氨酸(Phe)的浓度.结果:永州市部分地区15831例新生儿,初筛查出TSH＞10μIU/ml 720例,确诊CH 10例,发病率为1/1584,高TSH血症4例;PKU未检出.结论:新生儿疾病筛查的数量和新生儿疾病筛查的覆盖率是影响新生儿疾病筛查的发病率准确性的重要因素,只有扩大筛查范围,提高筛查率才能正确地反映全市的实际水平.