OBJECTIVE: To identify the gene mutation types of 4 suspected β-thalassemia patients in Yunnan Province, and to analyze the genotypes and hematological phenotypes. METHODS: Whole genome sequencing was performed on the samples of 4 suspected β-thalassemia patients from the Dai ethnic group in a thalassemia endemic area of Yunnan Province, whose hematological phenotypes were not consistent with the results of common thalassemia gene mutations. The mutations of β-globin gene clusters were confirmed by polymerase chain reaction (PCR) and Sanger DNA sequencing technology. RESULTS: The 3.5 kb deletion in β-globin gene cluster (NC_000011.10: g. 5224302-5227791del3490bp) was detected in 4 patients' samples, of which 1 case was also detected with HbE mutation and 1 case with CD17 mutation. These 2 patients displayed moderate anemia phenotype, while the two patients with only the 3.5 kb deletion presented with other mild anemia phenotype. CONCLUSION Heterozygous carriers with rare 3.5 kb deletion of the β-globin gene cluster may develop mild anemia, compound mutations of the 3.5 kb deletion with other mutations may led to intermediate thalasemia with moderate to sever anemia. In areas with a high incidence of thalassemia, suspected patients should undergo genetic testing to avoid missing or misdiagnosing rare mutations.
Humans ; beta-Globins/genetics* ; Multigene Family ; beta-Thalassemia/genetics* ; Mutation ; Genotype ; Sequence Deletion ; Phenotype ; Male ; Female

OBJECTIVE: To analyze the incidence of abnormal hemoglobin (Hb) in neonates in Guangzhou area, as well as the results of quantitative analysis of Hb in neonatal umbilical cord blood and genetic diagnosis of thalassemia in neonates with abnormal Hb; And to explore the hematological phenotypes and clinical characteristics of neonates with abnormal Hb and their parents, providing a reference for eugenics and childcare. METHODS: 650 neonates born at Guangdong Provincial People's Hospital who underwent Hb electrophoresis were included in this study. The results of routine blood test of umbilical cord blood , Hb electrophoresis and α-, β-thalassemia gene detection of the neonates were collected. The genotype distribution of thalassemia in the neonates was analyzed. Additionally, the abnormal Hb content of α and β variants was studied. Furthermore, the differences in hematological parameters between abnormal Hb neonates and normal neonates and α-thalassemia neonates, as well as between the parents of abnormal Hb neonates and normal adults were compared. RESULTS: Among the 650 neonates, 332 (51.08%) were diagnosed with thalassemia, including 235 cases of α-thalassemia (36.15%), 79 cases of β-thalassemia (12.15%), and 18 cases of compound αβ-thalassemia (2.77%). Among all the α-thalassemia genotypes, the most prevalent one was -- ^SEA/αα (48.94%), followed by -α^3.7/αα (20.00%), -α^4.2/αα (11.06%), and αα^CS/αα (8.94%). The four most common genotypes of β-thalassemia were β^CD41-42 (32.91%), β^IVS-Ⅱ-654 (26.58%), β^-28 (21.52%), and β^E (10.13%), respectively. 275 cases of abnormal bands were found in Hb electrophoresis of umbilical cord blood, with a detection rate of 42.31%. The abnormal Hb content of α-variant in the neonates was significantly higher than that of β-variant (P < 0.001). The levels of Hb, MCV, MCH, Hb A, and Hb F in neonates with abnormal Hb were lower than those in normal neonates, while the RDW-CV was higher than that in normal neonates, with statistical significantce (P < 0.05). The levels of RBC and Hb A in neonates with abnormal Hb were lower than those in neonates with α-thalassemia, while the level of MCH was higher than that in neonats with α-thalassemia, with statistical significance (P < 0.05). The levels of Hb, MCV, MCH, and Hb A in parents of neonates with abnormal Hb were lower than those in normal adults, while the RDW-CV was higher than that in normal adults, and the differences were statistically significant (P < 0.05). CONCLUSION The abnormal Hb content of α-variant in the neonates is significantly higher than that of β-variant in the neonates in Guangzhou, which can help to presume whether it is α chain or β chain based on the abnormal Hb content, providing a reference for globin gene sequencing. Meanwhile, analysis of various hematological screening-related indicators in neonates in the early stage is beneficial for early warning of the occurrence of abnormal Hb combined with thalassemia, reducing missed diagnoses to a certain extent.
Humans ; Infant, Newborn ; Genotype ; Hemoglobins, Abnormal/genetics* ; China/epidemiology* ; alpha-Thalassemia/epidemiology* ; beta-Thalassemia/genetics* ; Parents ; Female ; Male ; Fetal Blood

OBJECTIVE: To investigate the gene carrier rate and genotype distribution characteristics of thalassemia in the population of Kunming, and compare the differences of red blood cell (RBC) parameters between β⁺ heterozygous carriers, β⁰ heterozygous carriers and healthy population, as well as between different sexes of adults aged 18-45 years. METHODS: A retrospective analysis of 3 195 cases of thalassemia gene screened in the First Affiliated Hospital of Kunming Medical University from April 1, 2020 to March 31, 2022 was performed to detect 21 mutations of β-globin genes which was common in Chinese people using fluorescence PCR melting curve method. Patients with single heterozygous carrying β-thalassemia gene were divided into β⁺ heterozygote group and β⁰ heterozygote group, while the control group consisted of 219 healthy individuals. Four indices, including RBC, hemoglobin (Hb), mean corpuscular volume (MCV), and mean corpuscular hemoglobin (MCH) were collected from all β heterozygous carriers and 219 healthy people, and compared between β⁺ heterozygote group, β⁰ heterozygote group and control group, as well as between β⁺ heterozygous carriers, β⁰ heterozygous carriers and healthy population of different sexes aged 18-45 years. RESULTS: There were 688 cases confirmed thalassemia gene carriers, accounting for 21.53%. Among them, 322 cases were found to have β-globin gene mutations, including 145 cases of β⁺ heterozygote, 151 cases of β⁰ heterozygote, and 14 cases of β⁺ homozygotes as well as β⁺ and β⁰ dual heterozygotes. Additionally, 12 cases were found to have simultaneous mutation or deletion of β-globin and α-globin. The carrier rate of CD26 G>A mutation in β⁺ thalassemia was the highest, accounting for 57.9%, while in β⁰ thalassemia CD17 A>T was the highest, accounting for 46.4%. The erythrocyte parameters of 296 β heterozygous mutation carriers were compared with the normal reference interval, and it was found that 218 cases with RBC value greater than the highest value of reference interval, while 105, 281, and 269 cases with Hb, MCV, and MCH value less than the lowest value of reference interval, respectively. There were significant differences in the 4 erythrocyte parameters between β⁺ heterozygotes, β⁰ heterozygotes and healthy individuals (all P < 0.001), and further comparison between different sexes also showed significant differences (all P < 0.001). CONCLUSIONS The carrier rates of thalassemia gene and β-thalassemia heterozygote are both at high level in Kunming, and there are significant differences in the erythrocyte parameters between β⁺ heterozygous carriers, β⁰ heterozygous carriers and healthy individuals. When genetic counseling, it is necessary to inform and strengthen screening among adults of marriageable age to prevent birth of children with severe thalassemia.
Humans ; beta-Thalassemia/blood* ; Adult ; Heterozygote ; Male ; Female ; beta-Globins/genetics* ; Retrospective Studies ; Middle Aged ; Mutation ; Adolescent ; Genotype ; Erythrocytes ; Erythrocyte Indices ; Young Adult ; China ; Genetic Testing ; Asian People/genetics*

OBJECTIVE: To investigate the common genotypes and distribution characteristics of thalassemia in Guiyang region, and preliminarily analyze the rare mutations of globin genes in this area. METHODS: A total of 9 334 individuals who came to our hospital for thalassemia screening from June 2016 to February 2023 were included in this study. They were examined for common thalassemia mutations using PCR-based flow-through hybridization technology. Meanwhile, rare and unknown mutations were detected by Sanger sequencing. RESULTS: Among the 9 334 cases, 895 positive cases of common thalassemia were detected, with a positive rate of 9.59%. Among the positive samples, 565 cases (63.13%) were confirmed to be α thalassemia, of which the most common genotypes were αα/-α^3.7 (46.37%), followed by αα/--^SEA(26.55%) and αα/-α^4.2(10.62%); 310 cases (34.64%) were diagnosed as β thalassemia, with β^CD17/β^N (39.35%) being the most frequent genotype, followed by β^CD41-42 /β^N (31.29%) and β ^IVS-II-654/ β^N (12.90%). There were 20 cases (2.23%) of αβ complex thalassemia, mainly being αα/-α^3.7 combined with β^CD17 /β^N . Additionally, 8 cases of rare globin gene mutations were found by Sanger sequencing, including 7 mutation types. Among them, HBB: c. -137C> T (-87 C>T) was reported for the first time in Guizhou; HBA1 : c.*29C>T and HBB : c. 93-50C>T (IVS I-81C>T) were new mutations that had not been recorded in either the HbVar or IthaGenes database. CONCLUSION Guiyang region has a high incidence of thalassemia mutations, and these mutations are diverse and complex. Analyzing gene mutation types of thalassemia in this area can contribute to the prevention of the birth of children with severe thalassemia.
Humans ; Genotype ; Mutation ; beta-Thalassemia/genetics* ; alpha-Thalassemia/genetics* ; Thalassemia/epidemiology* ; Genetic Testing ; China/epidemiology* ; Male ; Female

OBJECTIVE: To explore the expression and function of microRNA-144 (miR-144) in β-thalassemia (β-thal). METHODS: The expression of miR-144 during the differentiation of murine erythroleukemia (MEL) cells and mouse embryonic liver-derived erythroid precursor cells was analyzed by real-time fluorescence quantitative PCR (qRT-PCR); The expression levels of miR-144 in peripheral blood and day-14.5 embryonic hepatocytes of wild-type (WT) and β-thal mice, as well as the expression levels of miR-144 in peripheral blood of β-thal patients, was also measured by qRT-PCR. The proportion of Ter119 and CD71 double positive cells in peripheral blood of mild and severe β-thal mice was analyzed by flow cytometry, and the expression levels of miR-144 in the peripheral blood of mild and severe β-thal mice and patients were compared; Bone marrow nucleated erythrocytes from WT mice and β-thal mice were sorted and the expression levels of miR-144 potential target genes were analyzed by gene chip. RESULTS: The expression levels of miR-144 were gradually increased during the directed differentiation of mouse MEL cells and embryonic hepatocytes to the erythroid lineage (r _MEL=0.97, r _{embryonic hepatocytes}=0.86); Compared with WT mice, the expression levels of miR-144 in peripheral blood and 14.5-day embryonic hepatocytes of β-thal mice were significantly increased (P < 0.05); Compared with healthy controls, the patients with β-thal showed an increased expression levels of miR-144 in peripheral blood (P < 0.05). Compared with mice and humans with mild β-thal, the expression levels of miR-144 in peripheral blood of those with severe β-thal were significantly increased (P < 0.05). The expressions of potential target genes of miR-144 in nucleated erythroid cells of the β-thal mice were significantly reduced compared to the WT group. CONCLUSION The expression level of miR-144 gradually increases in erythroid development, and compared with mild β-thal patients, the expression level of miR-144 in the peripheral blood is higher in severe β-thal patients. MiR-144 is expected to be an auxiliary diagnostic indicator for β-thal in clinical practice.
MicroRNAs/metabolism* ; beta-Thalassemia/genetics* ; Animals ; Mice ; Humans ; Cell Differentiation ; Hepatocytes

OBJECTIVE: To analyze the clinical features and laboratory characteristics of patients with cold agglutinin disease (CAD)/cold agglutinin syndrome (CAS) who were positive for acidified-serum lysis test (Ham test), and to compare them with Ham test negative CAD/CAS patients and paroxysmal nocturnal hemoglobinuria (PNH) patients, in order to provide references for the differential diagnosis of these diseases. METHODS: 53 patients diagnosed with CAD/CAS and 67 patients diagnosed with classic PNH in our hospital from January 2015 to December 2020 were retrospectively analyzed. The patients were grouped according to clinical diagnosis and results of cold agglutinin test (CAT), direct antiglobulin test (DAT), Ham test and PNH clone detection. The clinical and laboratory characteristics of each group were compared. RESULTS: The patients were grouped as follows: Ham^- CAD/CAS group, CAD/CAS patients negative for Ham test (n=36); Ham⁺ CAD/CAS group, CAD/CAS patients positive for Ham test (n=17); classic PNH group (n=67). Compared with the classic PNH group, the Ham⁺ CAD/CAS group had a higher median age (P =0.024), weaker positivity of Ham test, higher positive rates of CAT and DAT, and lower positive rate of PNH clone detection (all P <0.001). The proportions of patients with splenomegaly and cyanosis in Ham⁺ CAD/CAS group were significantly higher than those in classic PNH group (P =0.002 and P <0.001). Ham⁺ CAD/CAS group displayed lower red blood cell count (RBC) and lactate dehydrogenase (LDH) level (P =0.007 and P <0.001), and higher mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), and indirect bilirubin (IBIL) level (P =0.003, P =0.004 and P =0.006) than those in classic PNH group. The levels of serum complement C3 and C4 in Ham⁺ CAD/CAS group were lower than those in classic PNH group (P =0.001 and P <0.001). The positive rate of urinary occult blood in Ham⁺ CAD/CAS group was lower than that in classic PNH group (P =0.010). The clinical and laboratory characteristics of Ham⁺ CAD/CAS group were similar to those of Ham^- CAD/CAS group, except for median age, hemoglobin (Hb), MCHC, mean corpuscular volume (MCV), reticulocyte ratio (Ret), Ham test results, DAT positive types, and proportion of splenomegaly. CONCLUSION Some clinical features and laboratory indicators of CAD/CAS patients with positive results of Ham test are different from those of classic PNH patients, but relatively similar to those of CAD/CAS patients with negative results of Ham test. These results may provide a reference for differential diagnosis of related diseases.
Humans ; Anemia, Hemolytic, Autoimmune/blood* ; Retrospective Studies ; Hemoglobinuria, Paroxysmal/diagnosis* ; Female ; Male ; Coombs Test ; Diagnosis, Differential ; Middle Aged ; Adult

OBJECTIVE: To investigate the efficacy and safety of zanubrutinib in the treatment of autoimmune cytopenia (AIC) secondary to indolent B-cell lymphoma (iBCL). METHODS: A total of 23 patients with iBCL complicated with AIC who were admitted to our hospital from December 2019 to September 2023 were selected as the research subjects. All patients were administered zanubrutinib 160 mg, twice daily, and continued oral administration. The objective response rate (ORR) of AIC, the therapeutic effect on lymphoma, and the incidence of adverse reactions were observed. RESULTS: After a median follow-up of 20 (5 to 48) months, the median duration of response was 9 (interquartile range [IQR] 5-24)months. AICA efficacy assessment showed that there were 10 cases of complete remission (CR), 9 cases of partial remission (PR), and 4 cases of no response (NR), and the ORR was 82.6% (19/23) (95%CI : 61.2-95.0). Among them, for the 14 patients with autoimmune hemolytic anemia (AIHA), 7 achieved CR, 5 had PR, and 2 had NR. For the 4 patients with immune thrombocytopenia (ITP), 1 reached CR, 2 had PR, and 1 had NR. Regarding the 5 patients with Evans syndrome (ES), 2 achieved CR, 2 had PR, and 1 had NR. The assessment of lymphoma efficacy showed that there were 10 cases of CR , 7 cases of PR , 6 cases of stable disease (SD), and no progressive cases, with an ORR of 73.9% (17/23) (95%CI : 51.6-89.8). The main adverse reactions during the treatment were infection, hemorrhage, neutropenia, elevated lymphocyte count, rash, and anemia. Most of these adverse reactions were grade 1-2 and tolerable. No arrhythmia and hypertension occurred, and no deaths due to adverse reactions. CONCLUSION Zanubrutinib is effective and safe for AIC secondary to iBCL.
Humans ; Pyrazoles/therapeutic use* ; Lymphoma, B-Cell/complications* ; Pyrimidines/therapeutic use* ; Piperidines/therapeutic use* ; Female ; Male ; Anemia, Hemolytic, Autoimmune/etiology* ; Thrombocytopenia/etiology* ; Middle Aged ; Purpura, Thrombocytopenic, Idiopathic/etiology* ; Treatment Outcome ; Adult ; Aged ; Cytopenia

OBJECTIVE: To explore the characteristics of laboratory examination of secondary sideroblastic anemia (SA). METHODS: A retrospective analysis was conducted on the general information of a case of alcohol-induced secondary SA, and relevant domestic and foreign literature was reviewed to summarize the clinical characteristics of this disease. RESULTS: The patient was diagnosed with microcytic hypochromic anemia, with abnormal red blood cells detected, such as elliptocytes, target cells, and teardrop cells, etc. Iron overload and more than 15% of ring sideroblasts were detected by Perls' Prussian blue staining of bone marrow smear, while periodic acid-schiff (PAS) staining, EGR1 and SF3B1 gene tests were all negative, and rare genotype of Hong Kong α-thalassemia (possibly HKαα/αα or HKαα/-α^3.7) was observed. CONCLUSION Patient medical history and medication history, peripheral blood and bone marrow cytological morphology, genetic testing, cytogenetics, etc. are helpful for the diagnosis of secondary SA.
Humans ; Anemia, Sideroblastic/complications* ; alpha-Thalassemia/complications* ; Retrospective Studies ; Male

OBJECTIVE: To retrospectively analyze the genetic differences of thalassemia gene mutations among ethnic groups in Hechi, Guangxi. METHODS: A total of 15 595 whole blood samples of residents of Hechi from January 1, 2020 to June 30, 2023 were screened for thalassemia, and the Gap-PCR method and RDB-PCR method were used to perform genetic testing on the positive samples. Gene sequencing was performed on the samples with positive screening results but negative genotyping results. RESULTS: Among the 15 595 samples, 10 501 cases were screened positively, and 8 506 cases were thalassemia gene carriers among the positive samples, with a positive coincidence rate of 81.00%. Among them, there were 5 374 cases of α-thalassemia, 2 531 cases of β-thalassemia, and 601 cases of α+β compound thalassemia. A total of 13 mutant types were detected in α-thalassemia, including --^SEA (48.57%), -α ^3.7 (31.31%), α ^CS (8.57%) and -α ^4.2 (8.07%). A total of 17 mutant types were detected in β-thalassemia, mainly CD17 (48.27%) and CD41-42 (41.24%). The thalassemia gene carriers were mainly from the Zhuang (6 106 cases), Han (969 cases), Yao (793 cases), Mulam (275 cases), and Maonan (228 cases) ethnic groups. The comparison of constituent ratios within the above five ethnic groups demonstrated that there were differences in the proportions of -- ^SEA, -α ^3.7, α ^CS , and -α ^4.2 among the Zhuang, Han, and Yao ethnic groups (P < 0.005). The proportion of α ^CS in the Mulam ethnic group was not significantly different from -α ^3.7 and -α ^4.2. The proportions of -- ^SEA, -α^3.7, and α ^CS in the Maonan ethnic group were not significantly different. There were no significant differences in the proportion of CD17 and CD41-42 among the Han, Yao, Mulam and Maonan ethnic groups. The proportion of --^SEA was the highest in the Mulam ethnic group (56.68%), which was statistically different from 35.92% in the Maonan ethnic group. The proportion of -α ^3.7 was the highest in the Zhuang ethnic group (33.25%), and the difference was statistically significant compared to the Mulam ethnic group which had the lowest proportion (18.72%). The proportion of α ^CS was the highest in the Maonan ethnic group (27.46%), and the differences were statistically significant compared with other ethnic groups. The proportions of CD17 in the Zhuang and Maonan ethnic groups (50.79%, 55.68%) were higher than those in the Han (39.12%), Yao (39.63%) and Mulam (30.00%), and the differences were statistically significant. There was no significant difference in the proportion of CD41-42 among the above five ethnic groups. CONCLUSIONS The mutation type and distribution differences of genes causing thalassemia among main ethnic groups in the minority inhabited areas of Hechi, Guangxi, show the characteristics of ethnic differentiation. The result is helpful to develop a special prevention and control plan for thalassemia in line with the population distribution characteristics, and provide reference for revealing the genetic background and geographical distribution of thalassemia in this area.
Humans ; China ; beta-Thalassemia/genetics* ; Ethnicity/genetics* ; alpha-Thalassemia/genetics* ; Mutation ; Genotype ; Retrospective Studies ; Asian People/genetics* ; Thalassemia/genetics* ; Male

OBJECTIVE: To explore serological detection and blood transfusion strategies of mimicking antibodies, so as to provide appropriate transfusion strategies. METHODS: Detailed serological tests, including ABO blood group, Rh typing, antibody specificity, etc,were performed on two patients with autoimmune hemolytic anemia(AIHA). Meanwhile, the references about blood transfusion from mimicking antibody patients published from 1977 to 2024 in China and abroad were retrospectively summarized and analyzed. RESULTS: The patient 1 blood type was AB,CCDee and the antibody is mimicking anti-e, transfusion the e-negative red blood cells (RBCs) was effective. After two transfusions of e-RBCs, hemoglobin levels significantly increased from 48 g/L to 91 g/L, with complete resolution of hemolytic symptoms. The patient 2 blood type was O,CcDee, and the antibody was mimicking anti-c, the patient was diagnosed with AIHA and treated with hormone. No blood products were transfused during hospitalization, and his hemolysis was relieved. CONCLUSION Strictly grasping the indication of blood transfusion, blood transfusion should not be performed in the unnecessary conditions, and the corresponding antigen-negative RBC should be screened for transfusion in the necessay conditions.
Humans ; Blood Transfusion ; Anemia, Hemolytic, Autoimmune/therapy* ; ABO Blood-Group System ; Retrospective Studies ; Antibodies ; Male ; Blood Grouping and Crossmatching