Growth and development status of children with severe β-thalassemia and its correlation with serum HGB and SF levels in Bazhong area / 公共卫生与预防医学

Objective To analyze the growth and development status of children with severe β-thalassemia major (β-TM) in Bazhong area and its correlation with hemoglobin concentration (HGB) and serum ferritin (SF) level, and to provide a theoretical basis for the diagnosis and treatment of children with β-TM in Bazhong area. Methods A total of 292 children with β-TM admitted to Bazhong Central Hospital from January 2019 to December 2020 were selected. The Z-score method was used to evaluate the growth and development of the children. According to the growth and development of the children, they were divided into the normal group (normal growth and development, n=163) and delayed group (growth and development delay, n=129). Another 60 healthy children were selected as the control group. The levels of HGB, serum SF, free triiodothyronine (FT3), free tetraiodothyroxine (FT4), and thyroid stimulating hormone (TSH) were compared among the three groups of children, and clinical data such as age, sex and age of onset were collected from the case system. Univariate analysis and logistic regression were used to analyze the independent risk factors of growth and development delay in β-TM children. Pearson correlation was used to analyze the correlation between growth retardation and HGB and serum SF levels in β-TM children. Results The serum SF and TSH levels in the delayed group were significantly higher than those in the normal group and the control group, while the levels of HGB and serum FT4 were significantly lower than those in the normal group and control group (P2000 ng/mL were risk factors for growth retardation in β-TM children (P2000 ng/mL, the height, weight, and HGB and serum SF levels should be monitored regularly.

Risk factors for hemorrhagic cystitis in children with β-thalassemia major after allogeneic hematopoietic stem cell transplantation / 中国当代儿科杂志

OBJECTIVES@#To explore the risk factors for hemorrhagic cystitis (HC) in children with β-thalassemia major (TM) undergoing allogeneic hematopoietic stem cell transplantation (allo-HSCT).@*METHODS@#A retrospective analysis was conducted on clinical data of 247 children with TM who underwent allo-HSCT at Shenzhen Children's Hospital from January 2021 to November 2022. The children were divided into an HC group (91 cases) and a non-HC group (156 cases) based on whether HC occurred after operation. Multivariable logistic regression analysis was used to explore the risk factors for HC, and the receiver operating characteristic curve was used to analyze the predictive efficacy of related factors for HC.@*RESULTS@#Among the 247 TM patients who underwent allo-HSCT, the incidence of HC was 36.8% (91/247). Univariate analysis showed age, incompatible blood types between donors and recipients, occurrence of acute graft-versus-host disease (aGVHD), positive urine BK virus deoxyribonucleic acid (BKV-DNA), and ≥2 viral infections were associated with the development of HC after allo-HSCT (P<0.05). Multivariable analysis revealed that incompatible blood types between donors and recipients (OR=3.171, 95%CI: 1.538-6.539), occurrence of aGVHD (OR=2.581, 95%CI: 1.125-5.918), and positive urine BKV-DNA (OR=21.878, 95%CI: 9.633-49.687) were independent risk factors for HC in children with TM who underwent allo-HSCT. The receiver operating characteristic curve analysis showed that positive urine BKV-DNA alone or in combination with two other risk factors (occurrence of aGVHD, incompatible blood types between donors and recipients) had a certain accuracy in predicting the development of HC after allo-HSCT (area under the curve >0.8, P<0.05).@*CONCLUSIONS@#Incompatible blood types between donors and recipients, occurrence of aGVHD, and positive urine BKV-DNA are risk factors for HC after allo-HSCT in children with TM. Regular monitoring of urine BKV-DNA has a positive significance for early diagnosis and treatment of HC.

Optimization of β-globin Stable Expression Using the Third Generation Lentiviral Vector for β-thalassemia Therapy / 中国实验血液学杂志

OBJECTIVE@#To provide a research basis for a safe and effective cell therapy for β-thalassemia through optimization of HS4 region of the third generation lentiviral vector for stable expression of β-globin.@*METHODS@#The human β-globin HS4 region in the third generation lentiviral expression vector was optimized to construct the lenti-HBB, and the transcription and translation of β-globin gene were analyzed by RT-PCR and Western blot after the transduction of lenti-HBB in MEL cell line. Furthermore, the erythroid differentiation of CD34+ cells which were transduced lentiviral virus carrying human β-globin from normal human umbilical cord blood cells and peripheral blood cells of patients with β-thalassemia major were confirmed by colony formation assay, cell smear assay and flow cytometry. The safety and effectiveness of the optimized lenti-HBB were verified by NSG mouse in vivo test.@*RESULTS@#The human β-globin was expressed stably in the MEL cells, and CD34+ cells from health umbilical cord blood as well as PBMC from patient with β-thalassemia major transduced with lenti-HBB could be differentiated to mature red blood cells. The β-globin expression and differentiation in CD34+ cells were demonstrated successfully in the NSG mouse for about 35 months after post-transplant.@*CONCLUSION@#Stable β-globin expression through the optimization of HS4 from CD34+ in the third generation lentiviral vector is safe and effective for patients with severe β-thalassemia and other β-globin abnormal diseases.

Expression, functional mechanism and therapy application of long noncoding RNA in β-thalassemia / 中南大学学报(医学版)

β-thalassemia (β-thal) is one of the most common genetic diseases in the world, its pathogenesis is extremely complex and there is no effective treatment at present. The birth of children with moderate and severe β-thal brings economic pressure to families, social medical and health services. Long noncoding RNA (lncRNA) is a type of noncoding protein transcripts with a length greater than 200 nucleotides, which is involved in a variety of biological processes, such as cell proliferation, differentiation and chromosome variation and plays an important role in the epigenetic and post-transcriptional regulation of genes. It has potential value in the diagnosis, prevention and treatment of β-thal. LncRNA possesses the characteristics such as tissue specificity, cell specificity, developmental stage specificity, space-time specificity and disease specificity, and its complex interaction network has become a challenge to translate research results into clinical practice. Taking lncRNA as an entry point, in-depth understanding of the function of lncRNA in β-thal and explanation of its related regulatory mechanisms will provide theoretical basis for targeting treatment of β-thal, which can improve the diagnosis and treatment of β-thal.

Anaemia in pregnancy: prevalence and treatment response to various modalities: a prospective study

Background: Anaemia seen in pregnancy are largely preventable and easily treatable if detected in time, despite this, anaemia still continues to be a common cause of maternal and perinatal morbidity and mortality in India.Methods: A prospective observational study of 200 pregnant women with anaemia was carried out from Jun 2017 to December 2018 at a Tertiary care hospital with pan India population. Patients underwent clinical examination and laboratory tests to find out the severity and type of anaemia and were treated accordingly. Iron deficiency anaemia was treated with oral or intravenous iron therapy depending upon the hemoglobin concentration. Patients were followed up after 28 days of treatment and hemoglobin estimation was done to monitor the treatment response.Results: A total 36.49% pregnant women had hemoglobin less than 10 gm%. 151 out of 200 women had serum ferritin <12 ng/ml which indicates that iron deficiency anaemia is the commonest type of anaemia in pregnancy. Overall, out of 200 patients 5.5% patients were found to have hemoglobinopathies (β thalassemia trait). After 28 days of treatment mean increase in hemoglobin was 2.40 gm% and 4.24 gm% in patients receiving oral and intravenous iron therapy respectively.Conclusions: A total 36.49% pregnant women were found to have anaemia during pregnancy and iron deficiency anaemia is the commonest type of anaemia. Therefore, there is still a need for dietary counselling and health education in the community. 5.5% patients were found to have beta thalassemia trait which was detected only after conducting hemoglobin electrophoresis. Both oral and intravenous iron therapy are effective in treatment of iron deficiency anaemia but intravenous iron therapy results in a more rapid resolution of anaemia.

Effect of Chelation Therapy and Blood Transfusion on Trace Elements in Thalassemia Patients

Background: β-Thalassemia major is considered to be one of the most common inherited hemolytic anemia. Enhanced years of survival of thalassemia have led to unmasking related complications related to alterations in certain trace elements like magnesium, calcium, phosphorus, copper, zinc etc. Objective:Present study was conducted to evaluate the effect of iron chelation therapy and blood transfusion on certain trace elements (Magnesium, Calcium, Phosphorus, Copper, Zinc) in β-thalassemic patients on chelation therapy more than one year. Materials and Methods:In the present cross sectional study, 100 β-thalassemic patients receiving chelation therapy for atleast 1year were recruited from Civil Hospital Ahmedabad, Gujarat during February, 2017 to December, 2018 and equal number (n=100) of healthy subjects were taken as a control group in the age range of 8 to 15 years of both sexes (male & female). The levels of serum magnesium, calcium, phosphorus, zinc, and copper in serum were analyzed and results were correlated with normal healthy subjects. Results:A significant increase in serum copper (P≤ 0.01) and phosphorus (P≤ 0.001) were observed levels while a significant (P≤ 0.05) fall in magnesium, calcium and zinc levels recorded in β-thalassemic patients in comparison to healthy control subjects. Conclusion:Aforementioned observations suggested that fluctuations in the trace elements levels in β-thalassemic children receiving blood transfusion and iron chelation therapy could leads to different complications like hemolyzed red cells, infections & hemochromatosis renal damage, hypoparathyroidism etc. if remains untreated. Hence routine assessment of these elements is recommended for better management.

Ocular Manifestations of ß -Thalassemia Patients with the Use of Oral Iron Chelating Agents

Background: Ocular involvement in β-thalassemia major is very common. Iron chelators like Desferrioxamine and Deferiprone avoid systemic complications but chelate metals in retina. Objectives: 1.To study the relation of oral iron chelator (Deferiprone) on various ocular manifestations in β-thalassemia major patients. 2. To study the relation of serum ferritin with various ocular manifestations. Methods: 100β-thalassemia major patients out of those attending our thalassemia clinic were selected for the study as per our inclusion and exclusion criteria. They were divided into two major groups based on whether they were taking oral iron chelator (Deferiprone) or not. Detailed history, examination and investigations were done and recorded. Results: The study revealed that 52% of the patients had ocular involvement with 86.5% of them taking Deferiprone (p<0.0001), 13% had retinal pigment epithelium (RPE) degeneration with 92.3% of them on Deferiprone (p=0.003) and 18% had RPE mottling with 88.8% of them taking Deferiprone (p=0.001). Other ocular changes like lens opacity, disc hyperemia, best corrected visual acuity (BCVA) and venous tortuosity showed some difference between the two groups but that was insignificant. Further the study also showed that higher serum ferritin levels were significantly associated with ocular changes like decreased BCVA (p<0.001), RPE degeneration (p<0.001), RPE mottling (p<0.001) and venous tortuosity (p<0.025). Conclusion: Ocular changes in β-thalassemia major increases with greater duration of the disease and increased number of blood transfusions due to increased serum ferritin levels. Using iron chelators may reduce iron overload but they causechelator induced ocular involvement.

Health-related quality of life and its influencing factors in children with β-thalassemia major and their parents / 中华实用儿科临床杂志

Objective To evaluate the health-related quality of life (HRQoL) in children with β-thalasse-mia major and their parents,and to analyze its affecting factors.Methods PedsQLTM 4.0 generic core scale and a demographic questionnaire were used to assess HRQoL in 45 children with β-thalassemia major and their parents,which were between 5 and 12 years old,had received blood transfusion in Chengdu Women and Children's Central Hospital during 2016.Results The total summary score of patients' HRQoL was (74.58 ± 7.29) score,in which,the social functioning subscale score was the highest,and school functioning subscale score was the lowest.The total summary score of parents' HRQoL was (64.43 ± 11.54) score,and the difference was statistically significant (P ＜ 0.05).Parents' s core of the psychosocial health [(69.03 ± 12.24) score],the emotional functioning [(67.78 ± 12.67)score]and the School functioning [(57.92 ± 11.61) score]were significantly lower than those of children[(78.19 ±13.42) score,(83.75 ±9.05) score,(69.58 ± 10.30) score],and the differences were statistically significant(all P ＜0.05).The HRQoL of children was positively correlated with onset age of anemia and hemoglobin (Hb) level before transfusion (r =0.771,0.824,all P ＜ 0.01),but which was negatively correlated with iron chelation therapy (r =-0.573,P ＜ 0.01).In contrast,gender,frequency of blood transfusions during the previous 3 months,the type of blood transfusion and iron chelation treatment were not significantly related to HRQoL among these patients.All of these factors were not related to the HRQoL of their parents(P ＞ 0.05).A multiple regression analysis revealed that the HRQoL of children with β-thalassemia major was significantly correlated with onset age of anemia(P =0.005),Hb level before transfusion (P =0.026) and iron chelation therapy (P =0.000).Conclusions The HRQoL of children with β-thalassemia major and their parents were remarkably low.Comprehensive programs including social support,medical service and psychological care should be provided for these children and their parents.

Genotypic and phenotypic analysis of pregnant women with β-thalassemia gene carrying in Chengdu area / 国际检验医学杂志

Objective To investigate the differences in genotypes and phenotypic parameters of β-thalassemia gene carriers in pregnant women′s from Chengdu, Sichuan Province.Methods Totally 320pregnant women′s withβ-thalassemia gene from March 2016to June 2017in our hospital were selected.Routine blood tests, alkaline hemoglobin electrophoresis and routine analysis ofβ-thalassemia were performed on all the cases.Statistical analysis was performed on the data of each group.Results There were 306cases of heterozygousβmutations and 10types of mutations, among which 14cases ofα-thalassemia combined had 6types of mutations.The mutations of MCV, MCH, MCHC, and Hb in the routine blood tests of each group showed some differences.The incidence of abnormal bands was also different for each mutation, and the hemoglobin electrophoresis results ofβEM mutations contained abnormal bands.However, the clinical manifestations of CAPM mutations were not obvious and easily missed.Conclusion There is a certain regional specificity inβthalassemia gene carrying in Chengdu area.Targeted examination in the preliminary screening and prenatal diagnosis should be conducted so as to reducing the birth rate of children′s with severe thalassemia.

Research progress in hypogonadism of patients with β-thalassemia major / 国际儿科学杂志

Hypogonadism is one of the most common endocrine complications in patients with β-thalassemia major (β-TM).It can be clinically characterized by pubertal developmental delay,primary or secondary amenorrhea infertility that would significantly compromises the quality of life of patients with β-TM.Its pathogenesis is complex and may be related to long-term anemia,iron overload,decreased leptin levels,chronic liver disease,transplant pretreatment drugs and so on.Regular monitoring and active intervention are crucial for promoting adolescent development,sexual function maturation and retention of fertility in β-TM patients.However,there is no relevant guidelines and consensus in our country to guide clinicians on the followup of gonadal function in β-TM patients.This review aims to summarize the research progress in hypogonadism of patients with β-TM in order to improve the level of prevention and treatment.

Genomewide DNA Methylation Responses in Patients with β-Thalassemia Treated with Yisui Shengxue Granules () / 中国结合医学杂志

OBJECTIVE@#To examine the clinical effects of Yisui Shengxue Granules () in the treatment of β-thalassemia and explore its mechanism on DNA methylation levels.@*METHODS@#A randomized placebo-controlled double-blinded trial was conducted. Forty patients with β-thalassemia were recruited and distributed randomly by envelope method into an experimental group and a control group, 20 patients in each group. The patients were given Yisui Shengxue Granules in the experimental group and placebo in the control group (12 g/bag three times a day) during a 3-month intervention. Before and after 1, 2, and 3 months of treatment, peripheral intravenous blood was sampled, and blood parameters such as hemoglobin (Hb), red blood cells (RBCs), reticulocytes (Ret), and fetal hemoglobin (HbF) were analyzed. Mononuclear cells from 5 patients, who showed an obvious treatment effect, were isolated by density gradient centrifugation. DNA methylation was analyzed using an Affymetrix USA GeneChip Human Promoter 1.0 Array and Input-promoter 1.0.@*RESULTS@#Compared with pre-treatment, there was an obvious increase in Hb and RBCs counts after 1, 2, and 3 months in the experiment group (P<0.01 or P<0.05). Meanwhile, HbF increased from the 2nd to the 3rd month (P<0.05). In the control group, Hb and RBCs showed no obvioas change. After 3-month treatment, DNA methylation results from 5 patients revealed that there were 24 hypomethylated genes and 3,685 hypermethylated genes compared with pre-treatment. Genes of insulin-like growth factor 1 receptor (IGF1R) and Janus kinase 3 (JAK3) revealed the most relations with other genes (degree: 21) and genes of 1-phosphatidylinositol-4, 5-bisphosphate phosphodiesterase gamma 2 (PLCG2) and mitogen-activated protein kinase 10 (MAPK10) showed a stronger intermediary role (betweenness centrality=0.04).@*CONCLUSIONS@#JAK3 and MAPK10 are two key genes in bone marrow and the lymphatic system, and JAK3 is likely to be related to hematopoietic cytokines in the process of early hematopoiesis. (Registration No. NCT01549080).

Allogeneic hematopoietic stem cell transplantation for β-thalassemia major patients using a reduced-intensity conditioning regimen / 中华器官移植杂志

Objective To observe the efficacy and safety of reduced-intensity conditioning regimen used in allogeneic hematopoietic stem cell transplantation (HSCT) for children with β-thalassemia major.Methods We retrospectively analyzed the clinical data of 15 children with β-thalassemia major undergoing allogeneic HSCT with a reduced-intensity conditioning regimen from March 2013 to March 2017.Fifteen patients were diagnosed definitely,and the median age at transplantation was 5 years (range:3-6 years),including 11 with HSCT from unrelated donors (UDs),3 of HLA 8/10 matched and 8 of HLA10/10 matched.The remaining 4 patients out of 15 with HSCT were from related donors with HLA matched,3 donors were siblings and 1 was mother.All patients used a reduced-intensity conditioning regimen.The median mononuclear cell (MNC) dose and CD34 positive cell dose were 11.4 × 108/kg (range:4.8-20 × 108/kg)and 9.8 × 106/kg (range:5.9-27.2 × 106/kg),respectively.Graft-versus-host disease (GVHD) was prevented by cyclosporine A,methotrexate,MMF and ATGf.Results All 15 patients had successful engraftment.Median time to neutrophil and platelet engraftment was 12 days (range:9-21 days) and 15 days (range:10-25 days) respectively.Two patients developed grades Ⅱ acute GVHD and 4 patients developed chronic GVHD from unrelated donors,while there was no acute GVHD and 1 patient developed chronic GVHD from related donors.No patients suffered from serious transplantation-related complications,such as hepatic veno-occlusive disease (VOD),hemorrhagic cystitis,EB virus reactivation,CMV reactivation and hepatitis C,etc.The median follow-up time was 24 months (range:2-48 months).All patients were healthy and became transfusion-independent.Conclusion The reduced-intensity conditioning regimen proved to be safe and effective for children with β-thalassemia major given allogeneic HSCT.

2 Cases of Beta-thalassemia Minor in Korea / 임상소아혈액종양

In Korea, recent epidemiologic studies show that the incidence of β-thalassemia is increasing as the influx of South-East Asian population increases and molecular technologies develop. However, many patients are still misdiagnosed as iron deficiency anemia (IDA). All patients with microcytic anemia need to perform evaluation including reticulocyte index, Mentzer index, and iron studies. Considering the increasing incidence of β-thalassemia, hemoglobin beta globulin (HBB) gene sequencing should be performed if suspicious. In our cases, patients whose parents were both Koreans were confirmed to have β-thalassemia with a substitution in c1, ATG>GTG, and deletion of the HBB gene. In Korea, initiation condon ATG>AGG (20.9%) is most common mutation, followed by codon 17 (A>T) (17.6%), codon 121 (G>T) (12.1%), and so on. We report two cases of β-thalassemia diagnosed by genetic testing for microcytic anemia.

Correction of β-thalassemia mutant by base editor in human embryos

β-Thalassemia is a global health issue, caused by mutations in the HBB gene. Among these mutations, HBB -28 (A>G) mutations is one of the three most common mutations in China and Southeast Asia patients with β-thalassemia. Correcting this mutation in human embryos may prevent the disease being passed onto future generations and cure anemia. Here we report the first study using base editor (BE) system to correct disease mutant in human embryos. Firstly, we produced a 293T cell line with an exogenous HBB -28 (A>G) mutant fragment for gRNAs and targeting efficiency evaluation. Then we collected primary skin fibroblast cells from a β-thalassemia patient with HBB -28 (A>G) homozygous mutation. Data showed that base editor could precisely correct HBB -28 (A>G) mutation in the patient's primary cells. To model homozygous mutation disease embryos, we constructed nuclear transfer embryos by fusing the lymphocyte or skin fibroblast cells with enucleated in vitro matured (IVM) oocytes. Notably, the gene correction efficiency was over 23.0% in these embryos by base editor. Although these embryos were still mosaic, the percentage of repaired blastomeres was over 20.0%. In addition, we found that base editor variants, with narrowed deamination window, could promote G-to-A conversion at HBB -28 site precisely in human embryos. Collectively, this study demonstrated the feasibility of curing genetic disease in human somatic cells and embryos by base editor system.

Analysis on result of genetic screen and gene types of 10854 cases with thalassemia / 重庆医学

Objective To investigate the gene carrying rate,gene type and composition ratio of thalassemia among pre-pregnant population in Chongqing area.Methods A total of 1054 people were enrolled in the hospital from April 2014 to March 2016 for thalassemia screening.The content of screening included mean corpuscular volume (MCV),mean corpuscular hemoglobin (MCH) and hemoglobin electrophoresis.Thalassemia gene was examined in people with any abnormal term of screening result.Results In 10854 cases,1117 cases showed positive in thalassemia primary screening,and the positive rate was 10.29％.458 cases were tested positive of thalassemia gene,the carrying rate of thalassemia was 4.21％.In which,253 cases of pure a-thalassemia were tested.The carrying rate of α-thalassemia was 2.33％.The most common kind in α-thalassemia was--SEA whose constituent ratio were 52.17 ％.197 cases of pure β-thalassemia were tested,the carrying rate of β-thalassemia was 1.81％.The most common kind in β-thalassemia was CD17 (A→T),whose constituent ratio were 31.47 ％.11 cases were diagnosed with αβ-thalassemia.Conclusion Chongqing is high-prevalence area of thalassemia.It is important to conduct thalassemia genetic screen before pregnancy which plays a vital role in improving population quality and achieving prepotency.

Genotype analysis of α-thalassemia and β-thalassemia in child patients of Shenzhen region / 临床检验杂志

Objective To investigate the genotype and mutation frequency of thalassemia in child patients of Shenzhen region so as to provide evidences for the gene diagnosis and genetic counseling of thalassemia.Methods A total of 1 206 child patients suspected with thalassemia were retrospectively analyzed.The gene deletion of α-thalassemia was detected by Gap-PCR.The point mutations of α-thalassemia and β-thalassemia were determined by reverse dot blot(RDB)-PCR.The specimens suspected with HKαα and rare gene mutations were determined with nested PCR and gene sequencing,respectively.Results The detection rate of thalassemia was 76.9％ (927/ 1 206).Among them,α-thalassemia accounted for 40.5％ (489/1 206),and--SEA/αα was the most common gene mutation(75.1％);β-thalassemia accounted for 33.7％ (406/1 206),and the main IVS-2-654 (C→T) and CDM1-42 (-TCTT) heterozygous mutations accounted for 35％ and 32.5％,respectively.In addition,there were 32(2.7％) β-thalassemia patients with α-thalassemia mutation,1 patient with HKαα/ααQS,1 α-thalassemia patient with CD61 (AAG→TAG)/--SEA and 1 β-thalassemia patient with CD5 (CCT→C).Conclusion The are complicated gene mutation types and rare gene mutations of thalassemia in child patients of Shenzhen region.

MicroRNA expression in patients with β-thalassemia major / 国际检验医学杂志

Objective To explore microRNA (miRNA) expression patients with β-thalassemia major.Methods MiRNA differential expression were detected in β-thalassemia major by miRNA microarray analysis and quantitative real-time PCR.Results The results of miRNA array showed that 26 differential expression miRNAs were up regulated and 30 differential expression miRNAs were down regulated.Hsa-miR-618 which expression was up regulated and hsa-miR-103a-2-5p which expression was down regulated were selected for quantitative real-time PCR detection.It was showed that the expression tendency of hsa-miR-618 and hsa-miR-103a-2-5p were consistent.So the method of miRNAs array was reliable.The target genes of 17 miRNAs which were up regulated and 24 miRNAs which were down regulated were predicted by using database software.Conclusion It is notable that the differential expression of miRNAs in patients with β-thalassemia major.It will be afforded new direction and thinking for the mechanism research and disease treatment through the further research of the miRNAs regulation pathway in β-thalassemia major.

2 Cases of Beta-thalassemia Minor in Korea / 임상소아혈액종양

In Korea, recent epidemiologic studies show that the incidence of β-thalassemia is increasing as the influx of South-East Asian population increases and molecular technologies develop. However, many patients are still misdiagnosed as iron deficiency anemia (IDA). All patients with microcytic anemia need to perform evaluation including reticulocyte index, Mentzer index, and iron studies. Considering the increasing incidence of β-thalassemia, hemoglobin beta globulin (HBB) gene sequencing should be performed if suspicious. In our cases, patients whose parents were both Koreans were confirmed to have β-thalassemia with a substitution in c1, ATG>GTG, and deletion of the HBB gene. In Korea, initiation condon ATG>AGG (20.9%) is most common mutation, followed by codon 17 (A>T) (17.6%), codon 121 (G>T) (12.1%), and so on. We report two cases of β-thalassemia diagnosed by genetic testing for microcytic anemia.

A Novel Decrease of Mrp3 Protein in Liver of β-Thalassemic Mouse.

Background: Hepatocytes have a fundamental system of efflux proteins that protect cells from toxic insults. Unconjugated bilirubin at higher concentration is toxic to cells and its intracellular accumulation is limited by the induction of efflux proteins such as Mrp3. In vivo studies showed an induction of hepatic Mrp3 expression in response to non-hemolytic hyperbilirubinemia as a compensatory mechanism to reduce UCB toxicity. Study Design: In the present study, we analyzed the hepatic Mrp3 expression profile during hemolytic hyperbilirubinemia. We used β-thalassemic mouse and WT rodents treated with phenylhydrazine as an animal model of chronic and acute hemolysis, respectively. Results: Unexpectedly, Mrp3 protein was 75% down-regulated in β-thalassemic mouse although Mrp3 mRNA was normal. Mrp3 mRNA was significantly induced in PHZ treated animals while again; Mrp3 protein was 60% down-regulated. Conclusion: For the first time we observed a clear down-regulation for hepatic Mrp3 protein that linked to hemolysis, not to bilirubin. We hypothesize that a similar decrease for hepatic Mrp3 proteins is occur in hemolytic patients such as β-thalassemia.

The application of capillary electrophoresis in HbA1c and HbA2 detection in beta thalassemia patients / 中华检验医学杂志

Objective To investigate effects of β-thalassemia on 8 HbA1c measurement systems including Capillary electrophoresis and application of HbA2 values of samples with β-thalassemia from Capillary 2 Flex Piercing HbA1c measurement systems.Methods 338 samples without β-thalassemia ( include 80 α-thalassemia with --SEA/ααgenotype) and 225 samples with β-thalassemia were collected in Peking University Shenzhen hospital from January 2014 to August 2014.Samples withβ-thalassemia are all heterozygotes with normal α-globin and abnormal β-globin, and without combining other haemoglobinopathies.76 normal samples and 76 samples with β-thalassemia were selected from above all samples, HbA1c values of which were given by capillary electrophoresis ( Capillary Flex 2 Piercing), enzymatic ( Norudia N HbA1c ), immune transmission turbidity ( oneHbA1c FS ), chemiluminescent immunoassay (ARCHITECT HbA1c Reagent Kit), boronate affinity HPLC (Trinity Ultra2)and ion exchange HPLC (HA8160, VariantⅡTurbo and Variant Ⅱ BioRad 2.0), respectively.HbA1c values given by the eight systems were compared.338 samples without β-thalassemia and 225 samples with β-thalassemia were measured by using Capillarys 2 Flex Piercing HbA1c system and capillary electrophoresis hemoglobin system, the cut-off values of HbA2 from Capillary 2 Flex Piercing HbA1c measurement system for β-thalassemia was assessed by using ROC curve.Data statistics were done by using SPSS 19.0.Results The HbA1c values of normal samples were well correlated with the comparative system.The correlation coefficients for the 7 systems are all greater than 0.98.For samples withβ-thalassemia, the correlation coefficients are all greater than 0.98 except for Variant Ⅱ Turbo.The bias between normal samples and samples with β-thalassemia have no significant difference except for VariantⅡTurbo.HbA2 values from Capillary 2 Flex Piercing HbA1c measurement and capillary electrophoresis hemoglobin system were well correlated. The correlation coefficient is 0.993.When cut-off value of HbA2 from Capillary 2 Flex Piercing HbA1c measurement system for β-thalassemia is 3.38, sensitivity is 98.3%, specificity is 100% .Conclusions Samples withβ-thalassemia have no significant effects on HbA1c measurement syetems except for Variant Ⅱ Turbo.The Capillary 2 Flex Piercing HbA1c measurement syetem has the advantage of screeningβthalassemia by HbA2 while measuring HbA1c.