Spherocytosis Research Articles

Identification of a family with hereditary spherocytosis caused by a novel mutation of ANK1 gene

对1例不明原因的反复贫血患儿进行家系调查、临床和实验室检查及高通量测序分析，以明确其遗传学病因。患儿，女，5个月，出生2 d出现黄疸、不明原因的反复贫血。其父患有遗传性球形红细胞增多症，母亲健康。实验室检查：红细胞、血红蛋白减少，血涂片可见球形红细胞，基因检测示患儿及患儿父亲ANK1基因存在c.709C>T突变，为新发现的突变。提示ANK1基因c.709C>T突变可引起遗传性球形细胞增多症，是该患儿家系的遗传学病因。

Hereditary spherocytosis combined with malignant obstructive jaundice caused extreme hyperbilirubinemia: a case report

患者女性，54岁，遗传性球形红细胞增多症合并恶性梗阻性黄疸致超高胆红素血症。入院时总胆红素高达1 612.8 μmol/L，MRI示肝外胆管及胆总管扩张，经内镜逆行胰胆管造影(ERCP)置入支架解除梗阻。术后半个月因支架脱落，胆红素波动性上升，合并感染后溶血危象。再次经ERCP置入支架，经胆道镜观察狭窄段黏膜大致正常，经综合治疗患者情况稳定后行胆囊及脾切除术。术后病理未见明显异常，好转出院。随后因胆道感染再次入院经ERCP置入支架，于胆囊窝处穿刺病理提示腺癌。

Presentation of Rare Unstable Hemoglobinopathy in Mother and Son

Introduction Unstable hemoglobinopathies are rare genetic disorders that make up 10-25% of all hemolytic anemia disorders. It is often misdiagnosed for hereditary spherocytosis, aplastic anemia or iron deficiency anemia. Delay in diagnosis can alter management. We present two cases of Hemoglobin (Hb) Genova in a mother and child. Case Report Patient is a 41 year old female with history of anemia and scleral icterus as a child. She was initially diagnosed with hereditary spherocytosis and underwent splenectomy at the age of 19. Post-surgery her scleral icterus had resolved, …

Rapid screening of c. 166A>G in SLC4A1 gene by high-resolution melting analysis

Objective To explore the feasibility of a high-resolution melting (HRM) method for rapid screening of SLC4A1 mutation. Methods Two hereditary spherocytosis (HS) with a c. 166A>G heterozygous mutation of SLC4A1 confirmed by DNA sequencing and thirty healthy controls were selected for the study. The HRM primer was designed by Primer Premier 6.0 software. All of these samples were detected by LightCycler®480 and analyzed by HRM. Results The HRM analysis was able to detect the c. 166A>G heterozygous mutation of SLC4A1 effectively, and the specificity and sensitivity were both 100%. Conclusions The HRM analysis was appropriate for the detection of c. 166A>G in SLC4A1. It was an efficient, accurate and cost-effective molecular diagnosis method.(Chin J Lab Med, 2018, 41: 370-373) Key words: Spherocytosis, hereditary; Anion exchange protein 1, erythrocyte; High resolution melting

Analysis of Gene Mutation Related with Hereditary Spherocytosis in a Rhmod Patient of Chinese Han

Objective: Hereditary Spherocytosis (HS) is a cause of hereditary hemolytic anemia with mutations of one or more genes involved in the erythrocyte membrane. An Rhmod patient with mild anemia and spherical red blood cells was found in a small pedigree. His clinical symptoms were similar with that of hereditary spherocytosis. With the purpose of exploring whether the cause of mild anemia was caused by hereditary spherocytosis, HS-related genes, ANK1, SLC4A1, EPB42 were sequenced. Methods: Several exons of ANK1, SLC4A1 and EPB42 were amplified by polymerase chain reactions (PCR) and screened by direct Sanger sequencing. Results: a novel mutation of SLC4A1 (c.1552A>C) was detected. No mutations were found in ANK1 and EPB42 genes. Conclusion: The novel mutations of SLC4A1 might be related to HS. As the patient’s symptoms are similar with hereditary spherocytosis. Consequently, we speculate that anemia in the Rhmod patient may be related to HS.

Research Progress of Diagnosis of Hereditary Spherocytosis

Hereditary spherocytosis (HS) is the most common inherited disease of red cell membrane defects.Because of clinical characteristics,membrane protein defects and heterogeneity of genetic in HS,HS could be easily misdiagnosis.Currently,the progress of HS diagnosis is mainly focused on improving the sensitivity and specificity in detected of spherical red blood cells and cell membrane defects,but there is no single indicator can diagnose all HS.Eosin-5-maleimide binding test joint acidification glycerol dissolution test is the highest sensitivity combination test,and this method can be detected light and well compensated HS.Blood parameter analysis,blood smears and other tests are good for HS early screening.In this article,we review the historical origins,incidence,relationship between membrane protein defects and clinical manifestations,and the latest developments of laboratory tests in HS as follows. Key words: Spherocytosis, hereditary; Anemia; Anemia, hemolytic; Membrane proteins; diagnosis

Clinical and experimental studies of adult hereditary spherocytosis

To explore the clinical characteristic of adult hereditary spherocytosis (HS) and the efficacies of splenectomy. Six cases with adult HS were collected from July 2006 to July 2012. And their clinical features, laboratory profile and treatment were retrospectively analyzed. There were 2 males and 4 females with a diagnostic age of 21-67 years. Their chief complaints were symptoms of upper gastrointestinal tract. The characteristic tetralogy was anemia, hemolysis, splenomegaly and gallstone.Familial inheritance pattern was found in 2 patients. The laboratory examination results demonstrated anemia, reticulocyte and elevated indirect bilirubin in 6 patients. Three patients had positive urobilinogen. The osmotic fragility (OF) test were 2 positive and 4 negative ones. And 2 negatives turned positive after incubation. The acidified glycerol lysis test (AGLT50) was positive in all. Ultrasonography showed splenomegaly and gallstone. There were 3 abnormal types red cells in peripheral blood, densely-dyed red cells (35%-89%) , microspherocytes (14%-20%) and irregular-shaped red cells.Scanning electron microscope showed single or many acanthocytes or spikes in red cell surface.Red cell body was irregular dish or flower-shaped. Biconcave disc shape disappearance. Three patients had short bands 3, 4.1 or 4.2. The age of onset was late and clinical symptoms was lighter.Spectrin-ankyrin was deficient for another 2 patients. The age of onset was early and clinical sign serious.Splenectomy and cholecystectomy were performed in 3 patients. Their postoperative hemoglobin was normal during follow-ups. Clinical relief was achieved. Combined experiments can enhance the diagnostic rate of adult HS so as to avoid misdiagnosis. And the efficacies of splenectomy are definite.

Management of Spherocytosis

Management of Spherocytosis

Phosphatidylserine exposure in red blood cell in sepsis patients

Objective To investigate the morphologic change and phosphatidylserine (PS) exposure of erythrocytes in sepsis patients.Methods 30 healthy volunteers (control group)and 30 sepsis patients were enrolled in this study and were collected venous sampling.Monitoring included Wright's staining blood smear test,erythrocyte aggregation index and the ratio of PS exposure of erythrocytes.A flow-cytometric assay based on FITC-Annexin V was used to measure the PS exposure of erythrocytes.Results The morphological changes of red blood cells included acanthocyte,lachrymiform,rouleaux,spherocyte in sepsis patients,and the peripheral blood erythrocyte aggregation and aggregation index were significantly higher than that of the healthy control group (P＜0.05).The percentage of PS exposure of erythrocytes in sepsis patients were significantly higher than that of healthy volunteers (P＜0.001).Conclusion The PS exposure of erythrocytes were significantly higher in sepsis patients,and the morphology of red blood cells is obvious abnormal. Key words: Sepsis; Erythrocytes; Phosphatidylserine

Laparoscopic splenectomy: a single-center clinical study on 55 patients

Objective To study the safety and efficacy of laparoscopic splenectomy for splenic diseases.Method We retrospectively studied the outcomes of 55 patients who underwent laparoscopic splenectomy from May 2007 to December 2009.Splenic diseases included idiopathic thrombocytopenia purpura (n=11),autoimmune hemolytic anemia (n=6),hereditary spherocytosis (n=1),splenic lymphoma (n =1),splenic cyst (n=10),splenic angioma (n=5),vascular tumor of spleen (n=2),cirrhosis,portal hypertension and hypersplenism (n=9),cirrhosis and hyperplenism (n=9),and idiopathic splenomegaly (n=1).Results All patients underwent laparoscopic splenectomy,and there was no conversion to open surgery.The operation time (mean±S.D.) was (119.7±33.0) min.The intraoperative blood loss (mean± S.D.) was (83.8± 65.2) ml,and the postoperative hospital stay (mean±S.D.) was (5.7±1.1) days.One patient developed postoperative ascites,and 7 patients had drain fluid rich in amylase.There was no perioperatively death.Conclusion Laparoscopic splenectomy was safe and efficacious for splenic diseases. Key words: Laparoscopic splenectomy; Diseases of spleen

Autoimmune Hemolytic Anemia after Aplastic Crisis due to Parvovirus B19 Infection in a Patient with Hereditary Spherocytosis

eISSN 2093-6338 으로 알려져 있고 국내에서도 유전용혈 빈혈의 가장 많은 원인으 로 알려져 있다[2, 3]. 유전구형적혈구증과 같은 용혈성 적혈구 질 환에서는 질환의 경과 중 parvovirus B19 감염에 의한 적혈구계 전 구세포의 선택적 억제로 골수에서의 적혈구 생산의 감소가 발생되 어 혈색소가 급격히 감소하는 골수무형성위기(aplastic crisis)가 중 요한 합병증으로 잘 알려져 있다[4-6]. 유전구형적혈구증 환자에서 골수무형성위기는 7% 정도에서 발생하는 것으로 보고되었다[7]. 그러나 유전구형적혈구증에서 자가면역용혈빈혈을 동반한 경우 는 잘 알려져 있지 않다[8]. 자가면역용혈빈혈은 적혈구 항원에 대 해 자가항체가 발생하여 적혈구 파괴가 증가하여 빈혈을 야기하게 되는데 원발성으로 발생하거나 감염, 자가면역질환, 림프증식성 질 환 등에서 이차적으로 발생하기도 한다. Parvovirus B19는 드물게 자가면역용혈 빈혈을 야기한다[9-12]. 저자들은 유전구형적혈구증 으로 진단된 11세 남아에서 parvovirus B19 감염에 의한 골수무형 성위기 3개월 이후 회복기에서 자가면역용혈빈혈이 추가로 동반된 1예를 보고하고자 한다.

Surgical treatment of spleen-related blood diseases

Objective To explore the surgical indexes, therapeutic efficacy and prevention of complications in treatment of some blood diseases by splenectomy (including laparoscopic splenecto- my). Methods The clinical data of 27 patients with blood diseases receiving splenectomy in our hospi-tal were retrospectively analyzed. Results The effective rate of splenectomy was 100% in 4 cases of hereditary spherocytosis, 76.47% in 17 cases of ITP and low in 6 cases of Non-Hodgkin's lymphoma.The postoperative incidence of complications was 7.41% (2/27). Conclusion Splenectomy is of the first choice for treatment of hereditary spherocytosis. Meanwhile, it is effective for most of the patients with ITP. It is an important approach for comprehensive of malignant lymphoma. Proper choice of indications and perioperative management are helpful for decreasing incidence of postoperative complications. Key words: Splenectomy; Blood disease; Indication; Complication

Myopathy associated with spherocytosis and left ventricular hypertrabeculation/noncompaction

Myopathy associated with spherocytosis and left ventricular hypertrabeculation/noncompaction

原発性肺癌術後に発見された遺伝性球状赤血球症の１例

63歳男性, 集検の喀痰細胞診により発見された右B7入口部における扁平上皮癌症例である.平成13年3月胸腔鏡補助下S7区域切除を施行した.術後第2病日になりヘモグロビン値が急激に低下し黄疸が出現, 間接型優位の高ビリルビン血症を認め輸血を施行した.胸腔内出血, 下血等の症状もなく, 急激な貧血の進行は溶血性によるものが疑われた.諸血液学的検査所見並びに末梢血塗抹標本での多数の小型球状赤血球の確認により, 遺伝性球状赤血球症 (Hereditary-Spherocytosis, HS) と診断した.HSは, 我が国における先天性溶血性疾患において最も頻度が高く, その過半数が常染色体優性遺伝形式をとる.HSでは, 本症例の如く手術等の侵襲を契機にhemolytic crisis (溶血発作) を来すことがあり, 術前の詳細な病歴聴取, ならびに臨床症状, 検査成績からその存在を把握しておくことが重要であると考えられた.

Overwhelming Sepsis and Bleeding in a Previously Splenectomized Patient

The occurrence of overwhelming sepsis due to Streptococcus pneumoniae infection is a rare but well- known complication of splenectomy. Its occurrence 31 years postsplenectomy in a 35-year-old woman is the sub ject of this report. In spite of septic shock, disseminated intravascular coagulation, adult respiratory distress syn drome, gastrointestinal hemorrhage, and digital gangrene, the patient recovered with only minimal loss of parts of digits. The use of high-dose penicillin empirically in the emergency room along with liberal use of resuscitative fluids and dopamine may have been the major determi nants of her recovery. Intravenous corticosteroids, fresh- frozen plasma, and packed red blood cells appeared to be beneficial as well. A diagnosis of hereditary spherocyto sis was made following recovery. This case is discussed in the light of guidelines for prophylaxis of overwhelming sepsis in high-risk patients issued by the Centers for Dis ease Control of the National Institute of Health. Vacci nation with pneumococcal vaccine (23 valent) and men ingococcal vaccine is highly recommended for adults at high risk of overwhelming sepsis. This treatment should be repeated every 6 years. It is critical that the patient, the doctor, and emergency room physicians be aware of the importance of prompt aggressive antibacterial treat ment for any suspected infection.

Absence of close linkage between Hereditary spherocytosis (SPH) and 24 genetic marker systems including HLA and GM.

A total of 250 individuals belonging to 19 different families, identified through established propositi were simultaneously screened for hereditary spherocytosis (SPH), using stringent criteria, and 27 well-known polymorphic genetic marker systems. The segregation analysis indicated that the pattern of inheritance of SPH in these families, being autosomal and dominant, had a 100% penetrance. A statistical analysis, using the LIPED computer program of Ott (1974), revealed the absence of close linkage between SPH and ABO, ACP1, ADA, AK1, C3, DIA2, ESD, FY, GC, GLO1, GM, GPT1, HLA, HPA, JK, K, KM, MNS, P, PGD, PGM1, PGP, PI, and RH. Since an earlier study by other investigators had convincingly suggested a linkage between SPH and GM, we subjected the data to further analysis and found no significant heterogeneity in our recombination values of linkage between SPH and GM, or any of the other informative loci.

Neonatal Spherocytosis

Neonatal Spherocytosis

Effect of splenectomy of hepatic bilirubin clearance in patients with hereditary spherocytosis. Implications for the diagnosis of Gilbert's syndrome.

BRT and CBR were determined from studies of radiobilirubin kinetics in 14 patients undergoing splenectomy for hereditary spherocytosis. Studies were conducted both before and after the operation in order to assess the effects of the postsplenectomy fall in BRT on CBR. Splenectomy led to a doubling of the RBC-t1/2 from 13.0 +/- 1.7 (S.E.) days to 26.1 +/- 1.5 and a fall in BRT from 15.9 +/- 4.4 mg/kg/day to 4.3 +/- 0.9. In seven patients (group A) initially normal values for CBR were unaltered by surgery. In the remaining seven (group B), low preoperative values for CBR, suggestive of Gilbert's syndrome, uniformly improved after splenectomy, becoming normal in five patients. Nevertheless, family studies and reduced values for UDPGT activity supported the initial impression of concomitant Gilbert's syndrome in group B. These studies suggest the existence of a latent state for Gilbert's syndrome that, in some patients, may be unmasked by a hemolytic stress. As such, they have important implications for both pedigree analysis and incidence studies of this condition.

Invaginated plasma membrane of human platelets: evagination and measurement in normal and "giant" platelets.

Abstract A technique is described for estimating the amount of platelet plasma membrane. Addition of 80% by volume of distilled water to citrated human PRP converts platelets in the disc-form into large, spherical forms (spherocytes) without lysis in over 75% of these cells. Spherocyte formation is accompanied by about a twofold increase in mean surface area and a fourfold increase in the mean volume of the platelets. It is concluded that the large increase in surface area arises from an evagination of the platelet IM system. Spherocyte surface areas provide a measurement of total platelet plasma membrane surface area (including IM), which is in good agreement with recent electron microscopic estimations of platelet plasma membrane. This technique is utilized to measure plasma membrane of platelets obtained from donors with a giant platelet syndrome (MPS) in which discocytes are normal-sized and shape-changed platelets are abnormally large. The observation that MPS platelets have a normal amount of plasma membrane suggests that the abnormally large size of these platelets arises because of a subtle flaw in the mechanism which regulates membrane reorganization during platelet shape change. Osmotic spherocyte formation may be a rapid and useful technique for classifying giant platelet syndromes into (1) a group where the abnormally large platelet arises because of a defect in the way plasma membrane is organized and (2) a group where the abnormally large size is associated with increased plasma membrane. The significance of this kind of observation for the etiology of giant platelet syndromes is discussed.

Glycerol lysis time of incubated erythrocytes in the diagnosis of hereditary spherocytosis.

Abstract The median glycerol lysis time (GLT 50 ) of erythrocytes of normal subjects and patients with hereditary spherocytosis was compared before and after incubation for 24 hours at 37 °C. Normal erythrocytes, after incubation, had a median GLT 50 value of 24 seconds, with a range of 17 to 36 seconds (N = 61). A high linear correlation was found between GLT 50 values of fresh and incubated normal erythrocytes (r = +0.865). In hereditary spherocytosis, 16 out of 21 fresh specimens had GLT 50 values shorter than normal (median, 24 seconds; range 16 to 34 seconds), whereas after incubation all 22 of the specimens examined had GLT 50 values shorter than normal (median, 13 seconds; range 9 to 16 seconds). A modified glycerol reagent, producing a relatively slower rate of hemolysis, gave improved readability with incubated specimens, particularly in borderline cases. With the alternate method, the hereditary spherocytosis specimens had a median GLT 50 value after incubation of 18 seconds and a range of 12 to 22 seconds, substantially below the minimum normal value of 28 seconds. Blood specimens anticoagulated with EDTA and samples collected without anticoagulant (defibrinated blood and fingerstick specimens) had similar GLT 50 values and could be used interchangeably in this test, but heparinized specimens from the same subjects had significantly higher GLT 50 values.