Central Retinal Vein Occlusion Secondary to Paroxysmal Nocturnal Hemoglobinuria

Aims: To describe a Central Retinal Vein Occlusion Secondary to Paroxysmal Nocturnal Hemoglobinuria.Presentation of Case: A 25 years old, male, student, in regular follow-up in the Hematology sector due to Paroxysmal Nocturnal Hemoglobinuria in regular use of Eculizumab. He reports a month ago that he suddenly noticed, upon waking up in the morning, blurring and decreased visual acuity in her left eye.Discussion: Paroxysmal nocturnal hemoglobinuria (PNH) is an acquired disorder characterized by hemolysis, thrombosis, and bone marrow failure caused by defective expression of glycosylphosphatidylinositol-anchored (GPI-anchored) complement inhibitors. Most commonly, PNH is caused by the loss of PIGA function, which is necessary for GPI biosynthesis.Conclusions: Patients with hemolytic anemia, unexplained thrombosis, especially in uncommon sites, cytopenias and bone marrow failure syndromes, dysphagia, and unexplained abdominal pain should be screened for PNH. PNH patients can benefit greatly from the treatments available, with a reduction in the risk of serious sequelae and a considerable improvement in their quality of life. So that the patient continues to be followed up in the ophthalmology department, with decreased visual acuity ipsilateral to the lesion, with a relative afferent pupillary defect on that side.

Clinical characteristics of paroxysmal nocturnal hemoglobinuria (PNH) complicated with ischemic bowel disease / 中华内科杂志

Paroxysmal nocturnal hemoglobinuria (PNH) is a rare acquired hematopoietic stem cell disease. Gastrointestinal involvement is rarely seen in PNH. This study aims to analyze the clinical features in PNH patients complicated with ischemic bowel disease. Clinical date of 6 patients were collected at Peking Union Medical College Hospital from January 2010 to December 2020. The clinical manifestations, laboratory tests,imaging, endoscopic,and histopathological features and treatment were analyzed.Five in 6 patients were men, with a median age of 31 years old at onset. Most of disease course were recurrent episodes of chronic disease, with abdominal pain (5/6) and gastrointestinal bleeding (5/6). Laboratory examinations showed pancytopenia, reticulocytosis, elevated serum lactate dehydrogenase, high D-dimer and C-reactive protein levels in all patients. Multiple segments of small intestine were the most commonly involved and colon was also affected. Abdominal CT scan showed thickening and roughness or exudation of the intestinal wall (6/6), increased mesenteric density or “comb sign”(4/6), and cholestasis or gallbladder stones (5/6). Endoscopic manifestations included irregular shallow ulcers in the annular cavity (5/6), swelling mucosa with well-defined margins (6/6). Pathological biopsy revealed chronic inflammation of mucosa. The efficacy of steroids combined with anticoagulant therapy was better than that of steroids alone. Ischemic bowel disease in PNH patients is different from typical ischemic enteritis. Young patients, involvement of intestine with multiple segments are common characteristics. The anticoagulant is an essential agent for these patients.

Significance of paroxysmal nocturnal hemoglobinuria clone in immunosuppressive therapy for children with severe aplastic anemia / 中国当代儿科杂志

OBJECTIVES@#To study the association between paroxysmal nocturnal hemoglobinuria (PNH) clone and immunosuppressive therapy (IST) in children with severe aplastic anemia (SAA).@*METHODS@#A retrospective analysis was performed on the medical data of 151 children with SAA who were admitted and received IST from January 2012 to May 2020. According to the status of PNH clone, these children were divided into a negative PNH clone group (n=135) and a positive PNH clone group (n=16). Propensity score matching was used to balance the confounding factors, and the impact of PNH clone on the therapeutic effect of IST was analyzed.@*RESULTS@#The children with positive PNH clone accounted for 10.6% (16/151), and the median granulocyte clone size was 1.8%. The children with positive PNH clone had an older age and a higher reticulocyte count at diagnosis (P<0.05). After propensity score matching, there were no significant differences in baseline features between the negative PNH clone and positive PNH clone groups (P>0.05). The positive PNH clone group had a significantly lower overall response rate than the negative PNH clone group at 6, 12, and 24 months after IST (P<0.05). The evolution of PNH clone was heterogeneous after IST, and the children with PNH clone showed an increase in the 3-year cumulative incidence rate of aplastic anemia-PNH syndrome (P<0.05).@*CONCLUSIONS@#SAA children with positive PNH clone at diagnosis tend to have poor response to IST and are more likely to develop aplastic anemia-PNH syndrome.

Complement Inhibitors in Rare Diseases / 罕见病研究

The complement system is a self-protection mechanism of the human body. The abnormal activation of the complement system is involved in the occurrence and development of various diseases. The application of complement inhibitors in many rare diseases was a milestone in leading to the progress of such disease as paroxysmal nocturnal hemoglobinuria (PNH), atypical hemolytic uremic syndrome (aHUS), and others. Recently, the application of complement inhibitors has gradually expanded to other complement-related diseases. This review summarizes the literature on the current application of complement inhibitors in rare diseases and looks into the prospects of the application in the rare diseases.

Application of umbilical cord blood transplantation in the treatment of paroxysmal nocturnalhemoglobinuria / 中华器官移植杂志

Objective:To explore the feasibility and efficacy of umbilical cord blood transplantation (UCBT) in the treatment of paroxysmal nocturnal hemoglobinuria (PNH).Methods:From May 2014 to December 2019, clinical data were retrospectively reviewed for 7 PNH patients undergoing UCBT. The grades were severe ( n=6) and extremely severe ( n=1). The causes were primary PNH ( n=4) and PNH-aplastic anemia (AA) syndrome ( n=3). There were 5 males and 2 females with a median age of 29 (20-47) years, a median weight of 60(50-71) kg and a median time from diagnosis to transplantation of 62.5(7.7-171) months. All of them were accompanied by transfusion dependence. Myeloablative ( n=6) and reduced-intensity ( n=1) pretreatment was offered. The regimen of preventing GVHD was cyclosporine A plus short-term mycophenolate mofetil without ATG. The median number of input nucleated cells was 2.4(1.71-4.28)×10 7/kg and the median number of CD34+ cells 1.58(0.88-3.02)×10 5/kg. Results:Neutrophil and erythroid engraftment was obtained with a median neutrophil engraftment time of 17(15-21) days and a median erythroid engraftment time of 27. Engraftment time of 37(25-101) days for platelets >20×10 9/L and 62(27-157) days for platelets >50×10 9/L. The incidence of 100-day acute GVHD was 28.6%(95%CI 0-55.3%). The severity of GVHD was grade Ⅱ° acute ( n=2) and mild ( n=1). The median follow-up period was 13.5(3-71.4) months. Six patients survived while another with PNH-AA syndrome with iron overload died of gastrointestinal hemorrhage. The 2-year overall survival rate was 83.3%(95%CI 27.3-97.5%). Conclusions:With excellent engraftment and survival in the treatment of PNH, UCBT is indicated for patients without HLA full-match donor. PNH-AA syndrome with iron overload may be one of the important prognostic factors.

Hemoglobinuria paroxística nocturna y absceso hepático: asociación poco frecuente / Paroxysmal nocturnal hemoglobinuria and hepatic abscess: A little frequent association

Resumen: La hemoglobinuria paroxística nocturna es un trastorno adquirido de las células madre hematopoyéticas que se caracteriza por episodios de hemólisis intravascular. Aunque es una enfermedad poco frecuente, afecta en su mayor parte a adultos jóvenes, sin distinción de sexo. Comunicamos el caso de un paciente de 32 años de edad, que acudió a consulta con cuadro clínico de palidez, ictericia, hemoglobinuria y dolor en el hipocondrio derecho. El estudio de citometría de flujo de médula ósea reportó la ausencia de marcadores CD55 y CD59, indicativos del diagnóstico de hemoglobinuria paroxística nocturna, además de una imagen tomográfica hipodensa en el hígado compatible con absceso. En la bibliografía médica éste es el primer caso en el que se describe la coexistencia de estas dos afecciones.

Abstract: Paroxysmal nocturnal hemoglobinuria is an acquired disorder of hematopoietic stem cells characterized by episodes of intravascular hemolysis. Although it is a rare disease, it mostly affects young adults, regardless of sex. We present the case of a 32-year-old man with acute symptoms of paleness, jaundice, hemoglobinuria and pain in the right hypochondrium. The study of flow cytometry of bone marrow reported the absence of CD55 and CD59 markers, diagnostic indicators of nocturnal paroxysmal hemoglobinuria in addition to a hypodense tomographic image in the liver compatible with abscess. In the medical literature, this is the first case in which the coexistence of these two medical conditions is described.

Hemoglobinuria paroxística nocturna: revisión de la literatura / Paroxysmal nocturnal hemoglobinuria: literature Review

La hemoglobinuria paroxística nocturna (HPN) es una enfermedad clonal y adquirida causada por una mutación somática en el gen PIG-A que se encuentra en el cromosoma X y codifica una proteína involucrada en la síntesis del glicosilfosfatidilinositol (GPI), el cual le sirve como anclaje a muchas proteínas de la membrana celular produciendo mayor sensibilidad al complemento. Los distintos signos y síntomas que se presentan tienen gran impacto en la calidad de vida de los pacientes, por lo que un diagnóstico correcto es de vital importancia. Actualmente, la citometría de flujo multiparamétrica es la metodología de elección para detectar y seguir al paciente con HPN.

Paroxysmal nocturnal hemoglobinuria (PNH) is a clonal and acquired disease caused by a somatic mutation in the PIG-A gene found on the X chromosome and encoding a protein involved in the synthesis of glycosylphosphatidylinositol (GPI), which serves as anchoring to many proteins of the cell membrane producing greater sensitivity to complement. The different signs and symptoms that appear have a great impact on the quality of life of patients, so a correct diagnosis is of vital importance. Currently, multiparameter flow cytometry is the methodology of choice to detect and follow the patient with PNH.

Off-Pump coronary revascularization using bilateral internal thoracic arteries in a patient with paroxysmal nocturnal hemoglobinuria: a case report

Abstract Paroxysmal nocturnal hemoglobinuria (PNH) is an ultra-orphan disease. We report the first case in the literature of Off-Pump Coronary Revascularization Using Bilateral Internal Thoracic Arteries in a patient with paroxysmal nocturnal hemoglobinuria. A 36-year-old man came to the emergency department with acute non-ST elevation myocardial infarction (NSTEMI). He presented paroxysmal nocturnal hemoglobinuria diagnosed in 2016. Coronary angiography revealed tripple vessel disease. The conduits used for coronary revascularization were both internal thoracic arteries (left ITA-right ITA [LITA-RITA]). We consider that off-pump coronary artery bypass grafting (OPCABG) using Bilateral Internal Thoracic Arteries (BITA) can be safely performed with low in-hospital mortality and complications rates, even in patient with PNH.

Diagnóstico por citometría de flujo de la hemoglobinuria paroxística nocturna / Diagnosis of paroxysmal nocturnal hemoglobinuria by flow cytometry

Introducción: La hemoglobinuria paroxística (HPN) nocturna es una enfermedad clonal, adquirida y no maligna de la célula madre hematopoyética. En este padecimiento se encuentra afectado el anclaje a la membrana celular de moléculas como el CD55 y CD59, fundamentales en la regulación de la lisis mediada por el complemento. Por su elevada especificidad y sensibilidad, la citometría de flujo multiparamétrica (CFM) es el método de elección para el diagnóstico de esta enfermedad. Objetivo: Establecer un algoritmo diagnóstico de la HPN por CMF. Métodos: Se analizó una muestra de sangre periférica para CFM de un paciente con sospecha de HPN. El inmunofenotipaje celular se realizó con un panel de anticuerpos monoclonales dirigidos contra los antígenos que se expresan en la membrana citoplasmática mediante su anclaje al glicosilfosfatidilinositol. Las muestras se leyeron en un citómetro GALLIOS, Beckman Coulter y los datos obtenidos se analizaron con el empleo del programa informático Kaluza. Resultados: Se identificaron cuatro clones HPN. En los granulocitos se observó un clon HPN de aproximadamente 90 por ciento, con deficiencia en la expresión de CD16, CD24, CD55 y CD59. En los monocitos se observaron dos clones: (1) clon CD14_CD59_ y (2) clon CD14_CD59+ con tamaños clonales de 59,77 por ciento y 19,45 por ciento, respectivamente. En los eritrocitos se identificó un clon de 19,98 por ciento y de determinó el grado de afectación. Conclusiones: El algoritmo de análisis propuesto permite identificar las poblaciones celulares con clones HPN. Además, dichos clones pueden ser cuantificados en cuanto a tamaño clonal y expresividad de los antígenos dependientes de anclaje a glicosilfosfatidilinositol. Con la CFM se logra determinar con elevada sensibilidad el grado de afectación de los eritrocitos en la expresión de CD59 como medida directa de la susceptibilidad que experimentan a la lisis por el complemento(AU)

Introduction: The paroxysmal nocturnal hemoglobinuria (PNH) is a clonal, acquired disease and not malignant hematopoietic stem cell. In this condition, the anchor to the cell membrane of molecules such as the CD55 and CD59 is affected, This antigens are fundamental in the regulation of the complement-mediated lysis. By its high specificity and sensitivity multiparametric flow cytometry (MFC) is the goal standard for the diagnosis of this disease. Objective: To establish a diagnosis of PNH by MFC algorithm. Methods: A sample of peripheral blood of a patient with suspicion of PNH was analyzed by MFC. The cell immunophenotyping was carried out using a panel of monoclonal antibodies directed against antigens that are expressed in the cytoplasmic membrane through its the glycosylphosphatidylinositol anchor. The samples were read in a Cytometer GALLIOS, Beckman Coulter and the data obtained were analyzed with the use of the Kaluza software. Results: We identified four clones HPN. A HPN clone of approximately 90 percent, was observed in granulocytes with deficiency in the expression of CD16, CD24, CD55, CD59. In the monocytes were two clones: (1) CD14-CD59- clone and (2) CD14-CD59 + clone, with size clone of 59.77 percent and 19.45 percent, respectively. A clone of 19.98 percent was identified in erythrocytes and determined the degree of involvement of the same. Conclusions: The proposed analysis algorithm allows to identify cellular populations with clones PNH. In addition, these clones can be quantified in terms of size clonal and expressiveness of anchor to glycosylphosphatidylinositol antigen dependent. With the MFC is achieved with high sensitivity to determine the degree of involvement of the erythrocytes in the expression of CD59 as a direct measure of susceptibility undergoing lysis by complement(AU)

Outcomes of allogeneic hematopoietic stem cell transplantation in patients with paroxysmal nocturnal hemoglobinuria compared to paroxysmal nocturnal hemoglobinuria-aplastic anemia syndrome / 中华血液学杂志

Objective@#To compare the outcomes of allogeneic hematopoietic stem cell transplantation (allo-HSCT) for paroxysmal nocturnal hemoglobinuria (PNH) with paroxysmal nocturnal hemoglobinuria-aplastic anemia (PNH-AA) syndrome.@*Methods@#The outcomes of 46 patients who received allo-HSCT (16 PNH patients, 30 PNH-AA patients) from July 10, 2007 to June 2, 2018 were analyzed retrospectively. The conditioning regimen was busulfan, cyclophosphoramide, and ATG in haploidentical donors and unrelated donors. Patients with matched sibling donors were treated with the fludarabine, cyclophosphamide, and ATG regimen.@*Results@#There were no differences of baseline data between the 2 groups except gender distribution and the numbers of haploidentical donor transplantation. The median values of absolute nucleated cell counts were 10.58 (3.83-13.83) ×108/kg in the PNH group and 10.81 (3.96-33.40) ×108/kg in the PNH-AA group (P=0.668) . The median doses of CD34+ cells infused were 5.00 (3.14-8.42) ×106/kg and 3.57 (1.97-6.17) ×106/kg (P=0.002) , respectively. All patients obtained complete engraftment. The median time for myeloid engraftment were 11 (7-14) days in the PNH group and 12 (10-26) days in the PNH-AA group (P=0.003) . The median time for platelet engraftment were 13 (11-16) days and 18 (12-75) days (P=0.002) , respectively, after a median follow-up of 36 (4-132) months in the PNH group and 26 (4-75) months in the PNH-AA group (P=0.428) . There were no differences of incidence rates of acute graft-versus-host disease (aGVHD) , chronic GVHD and infection between PNH and PNH-AA groups (P>0.05) . No patient occurred early death and relapse. The estimated 3-year overall survival (OS) of PNH and PNH-AA groups were (100.0±0.0) % and (85.7± 6.6) % (P=0.141) , GVHD-free and failure-free survival (GFFS) were (100.0±0.0) %, (78.7±7.7) % (P=0.067) .@*Conclusions@#allo-HSCT is effective for patients with PNH and PNH-AA syndrome. The preliminary results indicate that myeloid and platelet engraftment in PNH group were faster than PNH-AA group. There were no differences in OS and GFFS between PNH group and PNH-AA group.

Expression and influencing factors of hepcidin in classical paroxysmal nocturnal hemoglobinuria / 中华血液学杂志

Objective@#To investigate the serum expression and influencing factors of hepcidinin patients with classical paroxysmal nocturnal hemoglobinuria (PNH) .@*Methods@#Retrospective analysis of 36 classical PNH patients from 2016.3 to 2017.3. Serum hepcidin concentration was measured by ELISA method. The relationship between serum hepcidin concentration and erythropoiesis and iron homeostasis parameters was evaluated.@*Results@#The median serum hepcidin level of 36 classical PNH patients was 32.03 (23.11, 118.48) μg/L, it was significantly lower than of 181.42 (106.80, 250.53) μg/L in 292 normal control subjects (z=-5.107, P<0.001) . The median serum hepcidin of 56.41 (44.60, 95.06) μg/L in PNH patients with normal ferritin was significantly lower than that in normal controls. The median serum hepcidin concentration 23.75 (21.77, 30.35) μg/L in iron deficiency PNH patients was lower than that in the normal ferritin PNH patients. However, the median serum hepcidin level of classical PNH with elevated ferritin patients 336.19 (304.19, 375.08) μg/L was significantly higher not only than that of normal ferritin and iron deficiency PNH ones, but also than that of normal control subjects. Regression analysis showed that serum ferritin, transferrin saturation and serum albumin level were independent influencing factors of serum hepcidin level in patients with classical PNH.@*Conclusion@#The decreased serum hepcidin level in patients with classical PNH was mainly influenced by iron metabolism factors.

Evaluation of Time and Temperature Stability of Paroxysmal Nocturnal Hemoglobinuria Cells by Flow Cytometry

BACKGROUND: Flow cytometry analysis of paroxysmal nocturnal hemoglobinuria (PNH) is significantly affected by the methodology used. The lack of data on the effect of age and refrigeration on PNH clone stability motivated us to study these aspects using flow cytometry. METHODS: Peripheral blood was collected from six patients, of which two presented with PNH. All samples were tested immediately and stored at room temperature (RT, 20–25℃) and at 4℃ for re-analysis at 24, 48, 72 hr and 7 days. Anti-CD59-fluorescein isothiocyanate (Beckman Coulter, USA) and anti-CD235a-phycoerythrin (PE; Beckman Coulter) were used to stain red blood cells (RBCs). Fluorescein-labeled proaerolysin (Cedarlane, Canada), anti-CD15-PE (Beckman Coulter), anti-CD24-PE-cyanin 5 (Beckman Coulter), and anti-CD45-PE-cyanin 7 (Beckman Coulter) were used to stain granulocytes. Flow cytometry was performed using a FC500 flow cytometer (Beckman Coulter). The effects of time and temperature were analyzed using generalized estimating equations. RESULTS: No significant differences in the gated percentage of RBCs and PNH clone size of RBCs were observed between the RT and 4℃ groups up to 7 days of testing. The percentage of gated neutrophils decreased with specimen age (P<0.001) and a better correlation with baseline was obtained at 4℃ than at RT (P=0.014). Neutrophil PNH clones were stable until 48 hr and 72 hr at RT and 4℃, respectively, and could not be analyzed at 7 days. CONCLUSIONS: RBC analysis was successfully performed up to 7 days. For neutrophils, testing within 48 hr is recommended, because the number of gated cells decreases significantly with age.

Comparison of hemolytic characteristics among paroxysmal nocturnal hemoglobinuria, autoimmune hemolytic anemia and hereditary spherocytosis / 中华血液学杂志

Objective@#To determine the valuable hemolytic characteristics in differential diagnosis of paroxysmal nocturnal hemoglobinuria (PNH), autoimmune hemolytic anemia (AIHA) and hereditary spherocytosis (HS).@*Method@#The clinical and hemolytic characteristics of 108 PNH patients, 127 AIHA patients and 172 HS patients diagnosed from January 1998 to April 2017 were compared.@*Results@#①Reticulocyte percentage (Ret%) of PNH patients [6.70% (0.14%-22.82%)] was significantly lower than that of AIHA [14.00%(0.10%-55.95%), P<0.001] and HS patients [11.83%(0.60%-57.39%), P<0.001]. The Ret% in PNH patients were significantly lower than those in AIHA and HS patients at the same levels of anemia, except for in mild anemia between PNH and AIHA patients. However, when comparing the Ret% between AIHA and HS patients, there was significant difference only in mild anemia [7.63%(1.87%-29.20%)% vs 11.20%(3.31%-22.44%), z=-2.165, P=0.030]. ②The level of TBIL in HS patients was significantly higher than that in AIHA and PNH patients [79.3 (11.2-244.0) μmol/L vs 57.6 (7.6-265.0) μmol/L, z=5.469, P<0.001; 79.3(11.2-244.0) μmol/L vs 26.2(4.6-217.7) μmol/L, z=-2.165, P<0.001], and the proportion of HS patients with TBIL more than 4 times the upper limit of normal (ULN) (64.1%) was significantly higher than that of AIHA (37.7%, χ2=19.896, P<0.001) and PNH patients (4.6%, P<0.001). ③The LDH level of PNH patients was significantly higher than that of AIHA and HS [1 500 (216-5 144) U/L vs 487 (29-3 516) U/L, z=-9.556, P<0.001; 1 500 (216-5 144) U/L vs 252 (132-663) U/L, z=-11.518, P<0.001], and the proportion of PNH patients with LDH more than 1 000 U/L (79.1%) was significantly higher than that of AIHA patients (13.0%, χ2=93.748, P<0.001) and HS patients (0, P<0.001). ④Splenomegaly occurred in 43.5% of PNH patients, including 16.0% with severe splenomegaly. In contrast, the occurrence of splenomegaly was 98.6% in AIHA patients and 100.0% in HS patients (P<0.001), and 63.0% of AIHA patients (P<0.001) and 90.4% of HS patients (P<0.001) were with severe splenomegaly. ⑤The prevalence of cholelithiasis in HS patients was up to 43.1%, significantly higher than that in AIHA patients (10.5%, P<0.001) and PNH patients (2.9%, P<0.001).@*Conclusion@#The comprehensive assessment of the five hemolytic characteristics is simplified, practical and efficient, with great clinical significance, providing specific indicators for differential diagnosis and efficient approach for making further work-up.

Purpura fulminans related to paroxysmal nocturnal haemoglobinuria: a case report and literatures review / 中华血液学杂志

Objective@#To improve the understanding of the rare clinical presentation and management of purpura fulminans (PF) in patients with paroxysmal nocturnal haemoglobinuria (PNH).@*Methods@#A case of PF occurring in PNH is reported, while the related literature review is conducted.@*Results@#A 49-year-old male patient suffered from one-week history of fever, greenish-brown colour urine, multiple well demarcated and painful purpura of the head and neck. He had been reported to have two thromboembolic events during the 22-year course of PNH. Skin biopsy displayed classic PF features. Laboratory testing showed a high PNH clone, intravascular hemolysis and coagulation system changes. After sufficient anticoagulation and short course of glucocorticoid therapy, the clinical conditions were improved correspondingly. During a follow-up period of 6 month, there was no recurrence of thrombosis.@*Conclusion@#PF should be considered in PNH patients with unexplained, quickly developed painful purpura. Extensive work-up should be performed to find out other potential thrombophilic risk factors after diagnosis of PF. Early diagnosis, adequate anticoagulation therapy and control hemolysis were essential to PF treatment occurring in PNH. The survival of patients and the qualities of life can be improved. The PNH clone detection is needed to evaluate the status of procoagulation and predict the risk of recurrent thrombosis.

Efficacy and safety of haploidentical hematopoietic stem cell transplantation for 17 patients with paroxysmal nocturnal hemamoglobinuria / 中华血液学杂志

Objective@#To explore the efficacy and safety of haploidentical hematopoietic stem cell transplantation (Haplo-HSCT) for paroxysmal nocturnal hemoglobinuria (PNH).@*Methods@#A total of 17 patients with PNH who received Haplo-HSCT from January 2013 to September 2017 were analyzed retrospectively.@*Results@#Of them, 4 patients had de novo PNH, 13 patients had aplastic anemia-PNH syndrome (AA-PNH). All patients received modified busulfan and Cytoxan (BuCy)-based regimens combined with anti-thymocyte globulin (ATG). Granulocyte colony-stimulating factor-mobilized bone marrow and peripheral blood stem cells were transplanted as graft. Prophylaxis for graft-versus-host disease (GVHD) was ciclosporin A+ mycophenolate mofetil (MMF)+short-term methotrexate (MTX). All patients were engrafted successfully. The median time of neutrophils to 0.5×109/L and platelets to 20×109/L was 12(10-15) days and 14(11-45) days, respectively. All of the 17 patients achieved full donor chimerism at 30 d after Haplo-HSCT. Seven patients developed grade Ⅱ-Ⅳ acute GVHD, and 4 chronic GVHD. Median follow-up time was 27.1 (8.6-60.4) months. Of the 17 patients, 15 survived and 2 died of severe pulmonary infection and transplant associated thrombotic microangiopathy. Three-year overall survival was (77.8±15.2)%.@*Conclusion@#Haplo-HSCT may be effective and safe for PNH patients who did not have matched donor.

Analysis of the role of paroxysmal nocturnal hemoglobinuria clones in acquired aplastic anemia in children / 临床儿科杂志

Objective To analyze the role of paroxysmal nocturnal hemoglobinuria (PNH) clones in children with acquired aplastic anemia (AA). Methods The relationship between the existence of PNH clones and clinical features in children with AA was retrospectively analyzed. The influence of PNH clones on the efficacy of combined immunosuppressive therapy (IST) of anti-thymocyte globuline (ATG) and cyclosporine A (CSA) was also observed. In addition, multiple factor analysis was used to analyze the main factors affecting the efficacy of AA. Results One hundred and forty-eight children with AA were enrolled, including 74 cases (50%) of granulocyte PNH clones positive, 68 cases (45.9%) of monocyte PNH clones positive, and 93 cases (62.8%) of total PNH clones (granulocytes and / or monocytes) positive. In 49 children having both granulocytes and monocytes PNH clones, the clone size of monocytes and granulocytes was 0.7% (0.4%-1.5%) and 0.2% (0.1%-0.7%), respectively, and the difference was significant (P<0.001) and there was a significantly positive correlation between them (r=0.65, P<0.001). According to the different PNH positive clones (monocytes, granulocytes, total), children were divide into three groups. And there were no differences in gender, age, concurrent infection, white blood cell count, hemoglobin concentration, platelet count, neutrophil absolute count, reticulocyte percentage in different PNH clones positive and negative groups (P>0.05). The group with monocytes PNH clones positive had a positive effect on the efficacy of IST (P=0.02). Multiple factor logistic regression analysis showed that the concentrations of hemoglobin and the positive PNH clones of monocytes were the main factors affecting the efficacy (P<0.05). Conclusions The high concentration of hemoglobin and the positive PNH clones of monocytes contribute the better effect of IST in children with AA.

Hemoglobinuria paroxística nocturna. el rol de la anhidrasa carbónica-i urinaria en la hemólisis intravascular / Peroxysmal nocturnal hemoglobinuria: the role of urinary carbonic anhydrase-I in intravascular haemolysis

La Hemoglobinuria Paroxística Nocturna (HPN) se caracteriza por hemólisis intravascular crónica mediada por complemento. Cuando se produce la hemolisis se libera a circulación Anhidrasa Carbónica- I (AC-I), una enzima que se halla en alta concentración en el eritrocito y por su bajo peso molecular filtra por el glomérulo. El objetivo del presente trabajo fue detectar la excreción de la AC-I en orina de pacientes con HPN por Electroforesis Bidimensional de Utilidad Clínica (2D UC), y compararla con otras causas de hemólisis, de origen renal y postrenal. Se evaluaron 8 pacientes con HPN sin tratamiento con eculizumab un inhibidor del C5 del complemento, y 5 de ellos postratamiento, 12 orinas de pacientes con nefritis lúpica y 10 orinas de pacientes con hemólisis postrenal. La AC-I puede estar presente en la orina, en los tres grupos, sin embargo la relación AC-I/Hemoglobina en la hemólisis intravascular está invertida en comparación con la hemolisis glomerular y post-renal. Los pacientes con HPN tratados con eculizumab no presentan AC-I, y sería de utilidad en el seguimiento de los pacientes tratados con el inhibidor del C5, para evidenciar posibles escapes hemolíticos.

Paroxysmal Nocturnal Hemoglobinuria (PNH) is characterized by chronic complement mediated haemolysis. In these conditions it might be expected that carbonic anhydrase-I (AC-I) would be liberated into the plasma and excreted in the urine, by its high concentration in the erythrocyte and low molecular weight. The objective of the present study was to detect the urinary excretion of AC-I from patients with PNH by wodimensional clinical utility electrophoresis (2D UC) and to compare it with other causes of renal and post-renal haemolysis. We evaluated 8 patients with PNH without eculizumab, a complement C5 inhibitor, 5 of them posttreatment, 12 urine of patients with lupus nephritis and 10 urine of patients with post-renal hemolysis. AC-I may be present in the urine, in all three groups, however, the AC-I/Haemoglobin ratio in intravascular haemolysis is reversed compared to glomerular and post-renal haemolysis. Patients with PNH treated with eculizumab do not have AC-I and would be useful in monitoring patients treated with the C5 inhibitor to evidence possible haemolytic leaks.

Hemoglobinuria paroxística nocturna / Paroxysmal nocturnal hemoglobinuria

ResumenLa hemoglobinuria paroxística nocturna es una anemia hemolítica crónica, adquirida, poco común, que afecta con igual frecuencia ambos sexos. Se manifiesta a cualquier edad y con mayor incidencia en países del sudeste asiático. Es el resultado de la expansión clonal no maligna de células progenitoras hematopoyéticas. Se caracteriza por anemia hemolítica intravascular, tendencia a la trombosis y un componente variable de insuficiencia medular.Se asocia a otras patologías hematológicas como anemia aplásica y síndrome mielodisplásico. La citometría de flujo es el método de elección para diagnóstico. El eculizumab y el trasplante de médula ósea alogénico son las únicas terapias efectivas.

Abstract:Paroxysmal nocturnal hemoglobinuria is a rare acquired chronic hemolytic anemia, which affects both sexes with equal frequency. It occurs at any age and more frequently in Southeast Asian countries. It is the result of non malignant clonal expansion of hematopoietic progenitor cells. It is characterized by intravascular hemolytic anemia, recurrent thrombosis and a variable component of bone marrow failure. It is associated with other hematologic disorders such as aplastic anemia and myelodysplastic syndrome. Flow cytometry is the method of choice for diagnosis. Eculizumab and allogeneic bone marrow transplantation is the only effective therapies.

Diagnostic Significance of Detecting Peripheral Blood Granulocyte CD55/CD59 and FLAER in Anemia and PNH / 现代检验医学杂志

Objective To investigate the clinical significance of CD55,CD59 and Aeromonas hydrophila toxin variant (FLAER) in the diagnosis of anemia and paroxysmal nocturnal hemoglobinuria (PNH).Methods Collected 30 healthy controls,22 cases of PNH,33 cases of aplastic anemia (AA),37 cases of iron deficiency anemia (IDA),45 cases of megaloblastic anemia (MA),30 cases of hemolytic anemia (HA) and 31 cases of myelodysplastic syndrome (MDS) from January 2009 to March 2017,CD55,CD59 and FLAER negative cell ratio of peripheral blood neutrophil of them were detected by multipa rameter flow cytometry.Results The detection rates of FLAER in PNH,AA and MDS groups were higher than those of CD55 and CD59,but there was significant difference in AA (x2 =7.759,5.518,P=0.005,0.019＜0.05).The average CD55,CD59 and FLAER deletion rate in PNH and AA group were significantly higher than those in normal control group and other groups (t=2.163～17.890,P=0.000～0.038＜0.05).The number of FLAER in PNH group was higher than CD59 and CD59 was higher than CD55 with the statistically significant difference (t=2.503 ～ 6.308,P=0.000 ～0.016＜ 0.05).Conclusion CD55,CD59 and FLAER have important value in the diagnosis of PNH and differential diagnosis with other anemia diseases,and can also be used to detect the presence of MDS and AA in patients with PNH.FLAER outperforms CD59,CD59 outperforms CD55.

Efficacy of eculizumab in paroxysmal nocturnal hemoglobinuria patients with or without aplastic anemia: prospective study of a Korean PNH cohort

BACKGROUND: Patients with paroxysmal nocturnal hemoglobinuria (PNH) often have concurrent aplastic anemia (AA). This study aimed to determine whether eculizumab-treated patients show clinical benefit regardless of concurrent AA. METHODS: We analyzed 46 PNH patients ≥18 years of age who were diagnosed by flow cytometry and treated with eculizumab for more than 6 months in the prospective Korean PNH registry. Patients were categorized into two groups: PNH patients with concurrent AA (PNH/AA, N=27) and without AA (classic PNH, N=19). Biochemical indicators of intravascular hemolysis, hematological laboratory values, transfusion requirement, and PNH-associated complications were assessed at baseline and every 6 months after initiation of eculizumab treatment. RESULTS: The median patient age was 46 years and median duration of eculizumab treatment was 34 months. Treatment with eculizumab induced rapid inhibition of hemolysis. At 6-month follow-up, LDH decreased to near normal levels in all patients; this effect was maintained until the 36-month follow-up regardless of concurrent AA. Transfusion independence was achieved by 53.3% of patients within the first 6 months of treatment and by 90.9% after 36 months of treatment. The mean number of RBC units transfused was significantly reduced, from 8.5 units during the 6 months prior to initiation of eculizumab to 1.6 units in the first 6 months of treatment, for the total study population; this effect was similar in both PNH/AA and classic PNH. CONCLUSION: This study demonstrated that eculizumab is beneficial in the management of patients with PNH/AA, similar to classic PNH.