Siriraj I Gγ(Aγδβ)0 -thalassaemia causing severe thalassaemia intermedia in compound heterozygous state with IVS1-1(G→T) mutation

@#The Siriraj I Gγ(Aγδβ)0 -thalassaemia is a novel mutation involving a 118kb deletion of the β-globin gene cluster. It was first reported in 2012 in two unrelated families from the southern part of Thailand. The carriers in the heterozygous state are clinically asymptomatic. Nonetheless, its complex interaction with other β-thalassaemia could give rise to different clinical phenotypes, ranging from mild thalassaemia intermedia to thalassaemia major. We report here a case of a six-year-old Malay boy, presented with pallor, growth failure and hepatosplenomegaly. His haemoglobin at presentation was 9.2g/dL with a mean cell haemoglobin of 22.6pg and a mean cell volume of 69.9fl. His peripheral blood smear showed features of thalassaemia intermedia. Haemoglobin (Hb) analysis revealed markedly raised Hb F (83%), normal HbA2 levels and absent HbA. Deoxyribonucleic acid (DNA) analysis showed compound heterozygous IVS1-1 (G→T) β-globin gene mutation and Siriraj I Gγ(Aγδβ)0 -deletion (genotype βIVS1-1/ β Siriraj I deletion). Both his father and elder sister are carriers of Siriraj I Gγ(Aγδβ)0 -thalassaemia while his mother carries IVS1-1 (G→T) gene mutation. Clinically, the patient is transfusion dependent on six weekly regime. To the best of our knowledge, this is the first reported case in Malaysia involving unique Siriraj I Gγ(Aγδβ)0 -thalassaemia and IVS1-1 (G→T) in a compound heterozygous state. In summary, detection of Siriraj I Gγ(Aγδβ)0 -thalassaemia is essential as this deletion can lead to severe disease upon interaction with a β-thalassemia point mutation as demonstrated in our case. The establishment of effective carrier screening and genetic counselling is important to prevent its adverse consequences.

Prevalence of beta thalassemia mutations in population of Gujarat using amplification-refractory mutation system–polymerase chain reaction

Background: Beta thalassemia is the most common genetic disorder in India. Its trait, coinheritance and mutations vary from mild to severe condition, resulting in thalassemia minor (heterozygous), intermediate and major depending upon many factors. The objective of this study was to find out the prevalence rate and the carrier of beta thalassemia in population of Gujarat using molecular genetic analysis of beta thalassemia patients by targeted mutation assay (ARMS-PCR).Methods: A total 105 samples for beta thalassemia were analysed for IVS 1-5 (G→C) and CD 15 (G→A) mutations. These two common mutations of thalassemia in Gujarat were carried out using amplification refractory mutation system–polymerase chain reaction (ARMS-PCR) and gel electrophoresis method.Results: A total 105 samples referred to us for molecular genetic analysis. The occurrence of positive mutations of IVS 1-5 (G→C) and CD 15 (G→A) were found in 48 and 15 samples respectively. The rest were negatives.Conclusions: Present study concludes that the prevalence rate of Beta thalassemia was widespread among the Gujarat population. The identification of IVS 1-5 (G→C) and CD 15 (G→A) mutations was carried out. The analysis revealed that, mutational patterns of IVS 1-5 (G→C) was the most frequent among other mutations in Gujarat region.

Analysis of clinical phenotype of a α2-globin gene mutation -IVS-Ⅱ-55 (T→G) / 临床检验杂志

Objective@#To identify a α-globin gene mutation-IVS-Ⅱ-55 (T→A) and analyze hematological characteristics of IVS-Ⅱ-55 (T→G) carriers. @*Methods@#The peripheral blood samples were collected from the members of five family and three sporadic IVS-Ⅱ-55(T→G) carriers for the analysis of RBC parameters and hemoglobin electrophoresis. Gap-PCR, PCR-RDB (reverse dot blot) and DNA sequencing were carried out for the identification of gene deletion and mutation of α-globin and β-globin. @*Results@#The results of RBC parameters of five infant probands which presented with microcytic hypochromic anemia were below the normal reference interval. One of the adult carriers of IVS-Ⅱ-55 (T→G) heterozygote alone presented with microcytic hypochromic anemia, and the others showed normal RBC parameters. The hematological phenotype index (MCV, MCH and HbA 2 ) of one adult carrying a compound heterozygote for IVS-Ⅱ-55 (T→G) and βCD27-28M/N were 65.0 fL, 20.3 pg and 5.8% respectively. The hematological phenotype index (MCV, MCH, HbA 2 and HbF) of one adult carrying a compound heterozygote for IVS-Ⅱ-55 (T→G) and SEA-HPFH were 81.9 fL, 26.5 pg, 3.0% and 29.0% respectively. The HbA 2 levels of all carriers of IVS-Ⅱ-55 (T→G) heterozygote alone were in normal range. No abnormal hemoglobin band was detectable on hemoglobin electrophoresis for all the carries. @*Conclusion@#The carriers of IVS-Ⅱ-55(T→G) heterozygote alone were asymptomatic. The phenotype of compound heterozygote for β-thalassemia was similar to that of β-thalassemia alone.

LARP2 DNA Methylation in Transfusion-dependent Haemoglobin E-Beta (HbE/β) and β-Thalassaemia Major Patients

Abstract@#Introduction: The large clinical spectrum of haemoglobin E-beta (HbE/β) thalassaemia leads to the investigation of complex mechanisms involved in erythropoiesis. DNA methylation in LARP2 is one of the potential epigenetic modifiers not fully explored in HbE/β and β-thalassaemia major. This study aimed to analyse DNA methylation profile and gene expression of LARP2 using peripheral blood (PB) in nucleated red blood cells (NRBCs) for the source of DNA of HbE/β- and β-thalassaemia major patients. Methods: PB were collected from 33 transfusion-dependent thalassemia patients from Hospital USM and Hospital RPZII, Kelantan, Malaysia. DNA methylation profile and gene expression of LARP2 were examined by bisulphite sequencing PCR and quantitative real-time PCR respectively. Results: Partial DNA methylation of LARP2 was observed in 43% (9/21) HbE/β- and 17% (2/12) β-thalassaemia major patients. LARP2 expression (1.49±26.60) in HbE/β-thalassaemia was not significant against normal controls and β-thalassaemia major (p>0.05). In contrast, LARP2 expression (6.8±16.42) in β-thalassaemia major showed a significant up-regulation against normal controls (p<0.05). The association of LARP2 expression and DNA methylation profile was statistically significant (p<0.001). LARP2 expression was down-regulated in 75% (3/4) HbE/β-thalassaemia patients with CD26/IVS1-5, in contrast to up-regulation of 80% (4/5) IVS1-5/IVS1-5 β-thalassaemia major patients. DNA methylation of LARP2 in these patients were either partially methylated or unmethylated in CD26/IVS1-5 and IVS15/IVS1-5 respectively. Conclusion: DNA methylation of LARP2 may act as an additional modifier to gene mutation especially involving IVS1-5 in HbE/β-thalassaemia. Homozygous IVS1-5 in β-thalassaemia major may contribute to different disease presentation compared to those involving CD26 in HbE/β-thalassaemia.

Surgical treatment of pulmonary atresia with intact ventricular septum / 国际儿科学杂志

Pulmonary atresia with intact ventricular septum(PA/IVS) is one of the complicated cyanotic congenital heart diseases with high mortality,which needs early intervention after birth.With further understanding of the disease and the development of surgery,the operative plan of the disease is gradually changing and some fundamental changes have taken place.The hemodynamic awareness of the disease is an important factor in the selection of surgery and the improvement of prognosis.In addition,it is also important for the selection of surgery and the improvement of prognosis to evaluate the development degree of the right ventricle comprehensively and accurately.At present,it is generally believed that the most effective treatment strategy can be obtained by classifying the development degree of the right ventricle in morphology.The surgical treatment has developed from early surgical pulmonary bypass to the interventional and hybrid surgery,which greatly improves prognosis and reduces surgical trauma.Among them,the pereutaneous pulmonary valve radiofrequency perforation plus balloon dilation and PDA stent implantation can be used as the preferred treatment for PA/IVS with dysplasia of the right ventricle.Particularly,PA/IVS can be cured with the interventional surgery alone for children with mild or even moderate dysplasia of the right ventricle.Hybrid surgery that significantly reduces trauma compared with traditional surgery is also an important choice for the areas where interventional surgery is relatively outdated.This article reviews and summarizes the progress of recognition and surgical treatment of PA/IVS,in order to provide reference for the clinical diagnosis and treatment.

Assessment of cardiovascular hemodynamics in gestational hypertension and preeclampsia.

Preeclampsia is a multisystem disease complicating 5-10% of pregnancies and remains in the top three causes of maternal morbidity and mortality globally. During pregnancy mean arterial pressure and vascular resistance decrease, while blood volume and basal metabolic rate increase resulting in increased cardiac output In hypertensive disorders of pregnancy there is currently no consensus on the systolic and diastolic parameters of cardiac function and the literature is conflicting regarding whether there is increased, decreased or any change in cardiac output. Women with a history of preeclampsia/eclampsia have approximately double the risk of early cardiac, cerebrovascular, and peripheral arterial disease, and cardiovascular mortality. This study was undertaken to evaluate cardiovascular hemodynamic alterations in hypertensive disorders of pregnancy in comparison with appropriately age, parity and gestational age matched control normotensive pregnancies. In women with preeclampsia cardiac work index and left ventricular mass index are increased as a result of increased workload on heart to maintain cardiac output against increased after load. Systolic function is well preserved. Diastolic function is reduced and those with global diastolic function are at increased risk of developing pulmonary edema. Advanced techniques like speckle tracking echocardiography can better identify those with compromised cardiovascular function.

A novel mutation in a case of dominant optic atrophy.

A 39‑year‑old healthy woman presented for decreased vision at distance and near for 4 years. She also noted a decrease in her color vision. Her best‑corrected visual acuities were 20/70 in each eye. Her visual fields were abnormal, and she had bilateral sluggish pupils, impaired color vision, and optic disc pallor. The magnetic resonance imaging of the brain, heavy metal screen, autoimmune work‑up, B12, B6, folate, erythrocyte sedimentation rate, rapid plasma reagin, and Lyme titer were all normal. Optical coherence tomography of the macula and electroretinogram were normal; the visual evoked potential was unrecordable in both eyes. She denied a family history of similar ocular issues, and genotyping of the OPA1 gene revealed a novel previously unreported mutation at IVS12+10T >C.

Polimorfismos FVII IVS7, FVII R353Q en el gen del Factor VII y FXIII Val34Leu y su asociación con el riesgo de infarto agudo del miocardio en pacientes costarricenses / FVII polymorphisms IVS7, FVII R353Q in FVII gene and FXIII Val34Leu and its associationwith the risk of acute myocardial infartion in costa rica patients

Los niveles aumentados de los factores de la coagulación como el FVII y FXIII se han asociado con infarto agudo del miocardio (IAM). Los estudios de biología molecular han permitido detectar varias mutaciones en los genes del FVII y FXIII que se han asociado al riesgo de enfermedad coronaria. Métodos: Se estudiaron 186 pacientes que sufrieron infarto agudo del miocardio y 201 controles sanos. Se determinaron los polimorfismos FXIII (Val34Leu);FVII, IV(S7), FVII (R353Q), según las técnicas descritas. Cabe destacar que esta investigación siguió los lineamientos de bioética. Resultados: La edad de los pacientes fue de 46,2 años (147 hombres/39 mujeres), y la de los controles fue de 46 años (141 hombres/60 mujeres). La prevalencia de las mutaciones obtenidas tanto en pacientes como controles fue: FVII IVS7 OR: 0,60 (0,26-1,38) p=0,193 y FVIIR353Q OR:0,81 (0,61-1,08) p=0,729; respectivamente. El fenotipo Leu/Leu tiene más elevada prevalencia en los casos controles que en los pacientes infartados, al hacer el ajuste de factores de riesgo cardiovascular, se demostró que este fenotipo es un factor protector para el desarrollo del IAM OR: 0,66 (0,47-0,93) p=0,01. Los factores de riesgo tradicionales fueron estadísticamente significativos. En el FVII, y en el FVII, y en el FVII IVS se encontraron nuevas variantes (4 y 8), no descritas previamente. Conclusión: El FXIII Val34Leu se presenta como factor protector contra el IAM, y ninguno de los polimorfismos del FVII se encontró asociado como factor de riesgo para IAM. El FXIII Val34Leu ha sido descrito como un facilitador de la activación del factor XIII, durante la fase final de la coagulación, incrementando y acelerando la estabilización de la fibrina, confiriendo más resistencia ante la fibrinólisis. Se incrementan el interés de este polimorfismo en el IAM y en especial para los pacientes que fueran sometidos a terapia antibrinolítica.

Increased levels of coagulation factors such as FVII and FXIII have been associated with acute myocardial Infarction (AMI).Molecular biology studies have identified several mutations in the genes of FVII and FXIII and observe its influence on thelevels of these and their possible association and risk of AMI.Methods: We studied 186 patients with documented AMI and 201 controls with no history of cardiovascular disease. Weperformed a case-control study. It was determined the FXIII Val34Leu polymorphisms, FVII IVS7, FVII R353Q, according tothe procedures described. This research followed the guidelines of institutional bioethics.Results: The mean age of patients was 46.2 years (147 men / 39 women), and controls was 46 years (141 men / 60 women).The prevalence of mutations obtained in patients as controls were: FVII IVS7 OR: 0.60 (0.26 to 1.38) p = 0.193 and FVIIR353QOR: 0.81 (0.61 to 1.08) p = 0.729 , respectively. It is observed that the phenotype Leu/Leu is more common in controls thanin patients, and after adjustment for cardiovascular risk factors, was shown to be a protective factor for the developmentof AMI OR: 0.66 (0.47 - 0.93) p = 0.01. The traditional risk factors were statistically significant. In FVII, were found in the FVIIIVS, new variants * (4 and 8), not previously described.Conclusion: FXIII Val34Leu was found as protector factor but neither FVII polymorphisms were associated as risk factors forAMI. FXIII Val34Leu, had been described as a facilitator of the activation of factor XIII, during the final stages of coagulation,increasing and accelerating stabilization of the fibrin, conferring mayor resistance to fibrinolysis. Increase the interest ofthis polymorphism in AMI and especially for patients who were subjected to antifibrinolytic therapy.

Comparative analysis of the recurrent mutations between Uigur and Han ethnic deaf group in Xinjiang region of China / 临床耳鼻咽喉头颈外科杂志

Objective:To investigate the recurrent mutations between Uigur and Han ethnic deaf group in Xinjiang region and determine the relationship between ethnicity and mutations.Method:DNA were extracted from peripheral blood of 125 deaf patients from Urumqi and Korla special educational schools in Xinjiang.Audiologic examinations showed that all patients had severe to profound bilateral sensorineural hearing hoss. The coding region of GJB2 gene, SLC26A4 and mitochondrial DNA target fragments were amplified by polymerase chain reaction(PCR).Mutations in GJB2 gene, SLC26A4IVS7-2 A>G, mtDNA 1494C>T and mtDNA1555 A>G were identified by sequencing analysis.Result:Allelic Frequency of the GJB2 35delG and SLC26A4IVS7-2 A>G mutations in Han deaf students were 7.4%and 10.1%,respectively, whereas not found in Uigur deaf groups.The difference was statistically significant. We did not find significant differences in GJB2 235 delC, 299-300delAT, mtDNA A1555G and C1494T allelic frequency between Uigur and Han students.Conclusion:Prevalence of the recurrent mutations between Uigur and Han ethnic deaf group in Xinjiang has a great diversity.

Relation of Xmn-1 Polymorphism & Five Common Indian Mutations of Thalassaemia with Phenotypic Presentation in b-Thalassaemia

Xmn-1 polymorphism is a known factor, which increases foetal haemoglobin production. Among, b thalassaemics in India five mutations are common. Disease severity was assessed based on age of presentation, age when received first transfusion and blood transfusion in ml/kg/year. Data was divided into three Xmn-1 categories, (+/+), (-/+), (-/-) and intergroup correlation was made. Mutations were divided into 2 groups, (group I) IVS 1-1 as one of its variables and (group II) without IVS 1-1. They were correlated. In Xmn-1 +/+ category 66.66% were diagnosed after one year of age, in mutations (group I) 53.57% had age of diagnosis after 1 year. 77.77% in Xmn-1 +/+ received their first blood transfusion after 1 year of age, in mutations (group I) 64.28% received their first blood transfusion after 1 year of age. About 66.66% patients in Xmn-1 +/+ category received blood <200ml/kg/year as against 72.22% in Xmn-1 -/- category. In mutations group I 57.14% received blood transfusion <200 ml/kg/year as against 68.18% in group II. It is concluded that the presence of Xmn-1 polymorphism and IVS 1-1 mutation leads to a milder phenotypic presentation causing a delay in onset of blood transfusions but dose not effect the amount of blood received /kg/year.

THE DIFFERENCE IN THE PREVALENCE OF THE IVS5-A/G OF THE TRANSFORMING GROWTH FACTOR-BETA3 GENE BETWEEN KOREANS AND ROMANIANS

Etiological role of polymorphism IVS+9C→G in hMSH2 gene in gastric cancers / 医学研究生学报

Objective: To investigate the etiological role of polymorphism IVS1+9C→G in hMSH2 gene in gastric cancers. Methods: A case-control study has been taken on subjects included 72 sporadic gastric, 71 familial gastric cancers and 126 healthy individual controls. Genomic DNA was extracted from peripheral white cell of all subjects. The polymorphism was detected by a PCR-based DHPLC analysis and verified by DNA sequencing. Results: The polymorphism IVS1+9C→G in hMSH2 gene was detected in 33.3%(42/126) Healthy individuals, 40.3%((29/72 ))sporadic gastric and 43.7%(31/71 )familial gastric cancers. Significant difference existed between cancers at young age (

Echocardiographic Parameters of Pulmonary Atresia with Intact Ventricular Septum(PA/IVS)

PURPOSE: To understand morphologic and hemodynamic variations in patients with pulmonary atresia with intact ventricular septum(PA/IVS), and to decide the best treatment modalities, we measured right ventricular volume, inflow, and outflow valvular annulus size in these patients and compared them with those of normal newborns. METHODS: Eight patients with PA/IVS diagnosed by echocardiography from January to December 2001 were enrolled in this study. Among the total eight patients, five were male and three were female. The mean age of patients was 6.9 days(1-34 days), and the mean body weight was 3,343 gm (2,970-4,000 gm). Ten fullterm newborn infants with sepsis or hyperbilirubinemia without heart disease were enrolled as a control group. Echocardiographic and Doppler studies using Acuson Aspen (7Mh probe) were recorded on super-VHS videotape and later on, with review mode. We measured volumes of right and left ventricles, aortic, pulmonic, mitral and tricuspid valvular annulus sizes using an installed program, and then these parameters were compared with those of the control group. RESULTS: Mean Z-value of tricuspid valvular annulus in PA/IVS was -3.69+/-2.80(-8.4--0.45), and tricuspid/mitral valvular annulus size ratio 0.68+/-0.15(0.43--0.84). The more the tricuspid/mitral valvular annulus size ratio, the more Z-value of tricuspid valvular annulus(P=0.003, r=0.885). Those patients who underwent pulmonary valvuloplasty(balloon or surgical) had a tendency toward larger volume of the right ventricle, more Z-value of pulmonic and tricuspid valvular annulus, and more tricuspid/mitral valvular annulus size ratio than those patients who underwent a shunt operation. CONCLUSION: Compared to a measurement of the volume of the right ventricle, measurements of tricuspid/mitral valvular annulus size ratio and Z-value of tricuspid valvular annulus may be easier and better parameters to decide the treatment method and to predict prognosis in PA/IVS patients.

Spnontaneous Regrssion of liver metastasis in Stage IV-S neuroblastoma after adrenalectomy: One Case Report

Prenatally diagnosed neuroblastomas have been reported in increasing numbers over the past several years. The vast majority are in favorable stages of the disease (stage I, II, IV-S). The authors experienced one case of stage IV-S neuroblastoma of the adrenal gland with liver metastasis, which regressed spontaneously after removal by adrenalectomy. This patient was noticed to have an abdominal mass at prenatal ultrasonography performed at 36weeks of gestation. This tumor was a neuroblastoma of the left adrenal gland with multiple liver metastases. Left adrenalectomy and liver biopsy were performed at 3 months of age. Thirty-eight months after surgery, an MRI demonstrated that the hepatic metastatic lesions had completely regressed without chemotherapy or radiation.

The effect of Large for Gestational Age on Asymmetrical Ventricular Septal Hypertrophy in the Newborn

PURPOSE: It has been known for a long time that infants of insulin dependent diabetic mothers are prone to develop macrosornia, organomegaly, hyperbilirubinemia, respiratory distress syndrome, hypoglycemia, hypocalcemia, septicemia and congenital anomalies in the neonatal period. And echocardiographic asymmetrical- ventricular septal hyper- trophy(ASH) has been observed in the newborn infants of diabetic mothers. The etiology of the ASH remains unknown, although fetal hyperglycemia and subsequent glycogen deposits have been postulated as contributing factors. Therefore, we have studied whether large for gestational age(LGA) has played an important role of developing ASH. METHODS: We compared echocardiographic findings in neonates of LGA and appro- priate for gestational age(AGA), who were admitted to the Department of Pediatrics, Chung Ang University Hospital from April 1994 to March 1997. RESULTS: 1) Mean LVED in LGA and AGA were 1.96+0.06cm and 1.94+0.04cm, respectively. 2) Mean LVPW in LGA and AGA were 0.37+0.05cm and 0.370.05cm, respectively. 3) Mean IVS in LGA and AGA were 0.400.09cm and 0.380.09cm, respectively. 4) Mean IVS/ LVPW in LGA and AGA were 1.09+0.12 and 1.040.17, respectively. There was no statistical significance between two groups in echocardiographic findings. CONCLUSION: There is no relation between LGA and ASH in the neonate. According- ly, we may not need to perform echocardiography LGA routinely.

A Case of Stage IV-S Neuroblastoma with N-myc Amplification and Coagulopathy / 대한소아혈액종양학회지

Neuroblastoma stage IV-S patients have frequent spontaneous remission and high survival rate. Many investigators have recommended minimal or no therapeutic intervention ; however, some patient do experience progressive disease and ultimately die of neuroblastoma. We experienced a case of stage IVS neuroblastoma with N-myc amplification and coagulopathy. This patient has treated with combination chemotherapy and radiation therapy, then remained disease free for 1 year on the follow up till March, 1997.

Cloning of human ? thalassemic mutation ? IVS II654(C→T) and establishment of its mammalian expression system / 中国病理生理杂志

AIM: To clone human ?-globin gene carrying a thalassemic mutation IVS II654(C→T) and establish a eukaryotic expression system for high-level expression of human ? IVS II654 gene in mouse erythroleukaemia(MEL) cells. METHODS: The fragments of human ? 654 gene isolated from the ? thalassemia patients homozygous for the ? 654 mutation were amplified by PCR, and cloned to plasmid pBGT51. Then, the human ? LCR and ? 654 gene were subcloned from plasmid pBGT51 to the stable mammalian expression vector pcDNA3.1+ together, and the MEL cells were transfected with this vector using commercially available cationic lipid FuGENE6. The MEL cells were induced for further maturation by DMSO and the expression of human ? 654 gene in the MEL cells was identified by RT-PCR. RESULTS: A mammalian expression system of human ? thalassemic mutation ?IVS II654(C→T) was established. CONCLUSION: The level and the reliability of expression of human ? 654 gene in the MEL cells in vitro are similar to that in vivo in human body. This may be a valuable gene therapy model for human ? thalassemic mutation ?IVS II654(C→T).