1例急性粒单核细胞白血病M_(4C)的MICM分型

目的报道1例急性粒单核细胞白血病M_(4C)的MICM分型,以提高对M_(4C)的认识。方法回顾性分析我院收治的1例M_(4C)的病历资料,并对其进行骨髓细胞形态学、细胞化学染色、骨髓活检、免疫表型、染色体核型、融合基因、二代测序等检查分析。结果患者骨髓细胞形态示:骨髓增生明显活跃,粒单核细胞占85.6%。细胞化学染色示:过氧化物酶(POX)染色部分弱阳性;特异性酯酶(AS-DCE)染色部分阳性;非特异性酯酶(α-NBE)染色部分阳性,且被氟化钠抑制;非特异性酯酶(AS-DAE)染色部分阳性,且部分被氟化钠抑制。骨髓活检示:骨髓增生极活跃,幼稚细胞弥漫性增生。免疫表型结果示:异常细胞群表达CD11B、CD64、CD56、cMPO、CD33、CD41、CD61、CD38和CD58,不表达CD13、CD34、CD117、CD7、CD123、HLA-DR、CD10、CD19、CD20、CD2、CD14、CD235、CD15、CD303、CD304、CD25、cCD79a、cCD3、cCD22、CD1a和TDT。染色体核型分析显示见克隆性异常t(9;11)(p22;q23),+mar。白血病43种融合基因筛查检出MLLT3-KMT2A融合基因阳性。二代测序检出NRAS、TET2基因突变阳性。诊断为AML(急性粒单核细胞白血病)M_(4C)伴t(9;11)(p22;q23);MLLT3-KMT2A。结论 M_(4C)既具有粒细胞系又具有单核细胞系的特征,细胞形态学表现复杂,其诊断必需结合MICM分型综合判断。     

细胞形态误诊为APL的CD56+急性单核细胞白血病1例

目的报道1例细胞形态误诊为急性早幼粒细胞白血病(APL)的CD56+急性单核细胞白血病,为临床上对其鉴别诊断提供依据。方法回顾性分析我院收治的1例形态似APL的CD56+急性单核细胞白血病患者的病历资料,并对其进行骨髓细胞形态、化学染色、免疫表型检测、染色体核型分析、融合基因等检查分析。结果患者骨髓细胞形态示:骨髓有核细胞增生明显活跃,以早幼粒细胞增生为主(占59.2%),考虑AML-M3;细胞化学染色示:髓过氧化物酶(POX)阴性,非特异性酯酶乙酸萘酯酶(NAE)和丁酸萘酯酶(NBE)双染色阳性,且阳性反应可被氟化钠抑制;免疫表型检测结果示:表达CD56,CD4dim,CD33,CD14,CD64,CD123,CD9,CD13,CD11b,MPOdim;部分表达HLA-DR,CD15;不表达CD7,CD117,CD34,CD16,CD19,CD22,CD20,c CD3,c CD79a,诊断为恶性幼稚单核细胞;染色体核型分析示:染色体核型正常(46,XY[20]);白血病43种融合基因筛查结果均为阴性。结论免疫表型分析可确诊形态特征不典型的无特定重现性遗传学异常的急性单核细胞白血病。     

急性粒单核细胞白血病与FAB分类

急性白血病的FAB分类中,对M_(1、2、3、5)的分型没有很大分歧,唯M_4粒单核细胞白血病较复杂,按照FAB分类大多数M_4患者的血及/或骨髓中,>20％的白血病细胞呈粒细胞系分化(过氧化物酶染色明显地比酯酶强,后者氟化钠不抑制),同时>20％的白血病细胞向单核细胞成熟(酯酶染色比过氧化物酶强,前者能被氟化钠抑制),这一亚型的白血病细胞在     

急性白血病患者八色流式免疫表型分析起始管方案的建立

目的建立急性白血病(AL)患者八色流式免疫表型分析起始管方案。方法用胞膜CD3(CD3)、CD19、CD10、CD34、CD45、胞浆CD79a(cCD79a)、髓过氧化物酶(MPO)和胞浆CD3(cCD3)等8种抗体建立八色流式染色方案。膜表面抗体直接染色;膜内抗体经固定破膜,再染色后上机检测。将3个血小板减少患者骨髓标本分别进行抗体的单色染色和缺一色染色;最后对17例确诊的AL初发患者标本进行检测。结果用单色染色来确定染色方案中各抗体的检测电压及荧光补偿;缺一色染色中,阳性细胞群较单色染色变化均<10%,表明方案中的各抗体相互作用小。17例AL初发患者中,6例急性B淋巴细胞白血病原始细胞均为CD34和CD19阳性,5例cCD79a阳性和4例CD10阳性;4例急性T淋巴细胞白血病患者均为cCD3阳性;6例急性髓细胞白血病均为CD34和MPO阳性;1例B+T混合表型AL患者CD34、cCD3、CD19、cCD79a及CD10均为阳性,MPO和CD3为阴性,此检测方案能够确定各类AL的细胞类型。结论建立了AL患者八色流式免疫表型分析起始管方案,操作简便快速,适用于临床检测。     

母细胞性浆细胞样树突状细胞肿瘤临床与实验室特点分析

目的 对1例母细胞性浆细胞样树突状细胞肿瘤(BPDCN)患者的临床表型、形态学特点、免疫学表型、染色体核型等进行分析,以提高临床和实验室人员对此类病例的认识。方法 分析1例BPDCN患者的临床表型、骨髓细胞形态学、免疫表型与遗传学结果,并结合文献总结BPDCN的临床特征。结果 BPDCN患者骨髓涂片瑞氏-吉姆萨染色结果:骨髓增生明显活跃,原始细胞37.5%,该类细胞中等大小;核圆形、椭圆形或不规则形,染色质细致,核仁隐显不一;胞质量中等,灰蓝色,无颗粒,部分细胞胞质中有沿细胞膜分布的空泡和伪足;原始细胞过氧化物酶(POX)染色阴性;过碘酸-希夫(PAS)染色为红色颗粒状阳性。骨髓流式细胞学检测结果:异常细胞群占有核细胞的13.15%,强表达HLA-DR、CD56、CD123,表达CD33、CD7、CD4,部分表达CD38,不表达cCD3、cCD22、cMPO、CD3、CD10、CD34、CD117。染色体核型分析结果为46,XY(20)。骨髓细胞形态学和免疫表型结果均符合BPDCN的诊断标准。结论 临床表型、细胞形态学和免疫表型对于诊断BPDCN都有重要意义,其中免疫表型必不可少。     

腹膜后碰撞瘤1例

正患者男性,83岁。2014年1月CT发现脾大及腹腔实质性占位病变(考虑来自腹膜后)。同年12月查血常规:白细胞数 42.65×109/L,淋巴细胞 38.99×109/L,单核细胞 0.61×109/L。骨髓细胞涂片检查示骨髓有核细胞极度活跃,全片以淋巴细胞增生为主(占95.6%),粒系、红系增生受抑,成堆血小板可见。流式细胞检查示CD19、CD20、CD22、CD5、C11c、HLA-DR均阳性。结合患者病史、临床症状诊断为慢性淋巴细     

单核细胞白血病的分类

单核细胞白血病的分类作为独立的一个类型长时期被怀疑,许多人不承认此型白血病。目前单核细胞在骨髓中发生的概念已被大多数学者所接受。近十年来的研究,发现了高活性的非特异性酯酶并可被氟化钠抑制试验和大量溶菌酶产物,给单核细胞提供了本质。目前可将单核细胞白血病分为四个基本型:①急性单核或原单核细胞白血病;②急性粒-单核细胞白血病;③慢性单核细胞白血病;④慢性粒-单核细胞白血病。发病率以第一型最少见,第二型最多。急性单核细胞白血病的鉴定和分类除形态标准外,更重要     

流式细胞仪和免疫组织化学法测定骨髓增生异常综合征、急性髓系白血病患者和正常人骨髓中的CD34阳性细胞的比较

急性髓系白血病(AML)患者骨髓白血病细胞上CD34高表达预示着短生存期和/或对大剂量化疗疗效差,同样骨髓增生异常综合征(MDS)患者骨髓及外周血中CD34阳性细胞增多与短生存期和转化为AML高危险性相关。作者使用流式细胞仪(FC)测定新鲜骨髓细胞悬液中CD34阳性细胞和免疫组织化学法(IH)测定石蜡包埋骨髓样本中CD34阳性细胞,并进行二种方法的比较。 受检患者分为三组:MDS组16例,其中难治性贫血3例,环状铁粒幼细胞性贫血5例,原始细胞过多性难治性贫血3例,转化中原始细胞过多性难治性贫血2例及慢性粒单核细胞白血病3例;AML组12例(M1     

急性髓细胞性白血病微分化型的临床研究

目的探讨急性髓细胞性白血病微分化型的诊断和治疗。方法骨髓涂片作细胞化学分析。抗凝骨髓液用流式细胞单克隆抗体直接免疫标记技术检测白血病细胞表面相关抗原分化群 ,咐醇酯诱导分化试验 ,短期培养法直接法G显带分析白血病细胞染色体组型。用TA/HA/IdaA等方案化疗 ,达完全缓解 (CR)后作异基因外周血干细胞移植。结果骨髓增生极度活跃 2例 ,明显活跃 1例 ,活跃 1例 ,均未见Auer小体及嗜天青颗粒。早期微分化细胞NEC均 >90 % ,过氧化酶染色均阴性 ,糖元染色 2例弱阳性 ,2例阴性 ,非特异性酯酶染色 2例阳性 ,氟化钠不抑制。 1例咐醇酯诱导分化试验强阳性。免疫表型 3例CD+ 1 3,1例CD+ 1 5,1例CD+ 38,1例CD+ 1 1b,4例MPO弱阳性 ,CD1 9、CD2 均阴性 ,2例CD+ 7,白血病细胞染色体组型未见异常。 2例化疗达完全缓解 ,其中 1例现已存活 2 8月余 ;1例存活 1 9月 ,因合并肥厚心肌病放弃治疗 ,复发死亡。另 2例化疗无缓解 ,死亡 1例 ,自动出院 1例。结论细胞化学、细胞免疫表型是诊断急性髓细胞性白血病微分化型的重要依据 ,用AML方案治疗ANLL M0 有效。     

氟化钠抗非特异性酯酶NAE活性敏感度测定的研究

[目的]本文通过氟化钠对非特异性酯酶NAE活性抑制试验，探讨其在疾病诊断中的应用价值。[方法]使用10个不同浓度的氟化钠，对2060份骨髓涂片标本进行抑制前后非特异性酯酶活性强度观察。采用王氏改良α-醋酸奈酯酸测定非特异性酯酶（NAE）组化法，观测NAE的阳性率和阳性指标。[结果]不同浓度的氟化钠均不能完全抑制各类骨髓细胞所呈现的不同强度的NAE反应。仅有部分细胞呈现轻度或中度部分抑制。氟化钠处理前后两组的NAE活性阳性率和阳性指数无显著差异（P〈0.05）。[结论]通过氟化钠抑制前后NAE组化染色的大量实验结果，说明氟化钠抑制试验对骨髓细胞种类和白血病类型鉴别无大参考价值。本实验结果也为重新评估NAE的价值提出了新的观点和实践依据。     

湖南地区成人血清IgG亚类水平调查及影响因素分析

目的调查湖南地区成年人血清IgG亚类的浓度水平,探讨年龄、性别及生活方式等因素的影响。方法用免疫散射比浊法测定170例体检者血清中IgG_1、IgG_2、IgG_3、IgG_4和IgG浓度。结果血清IgG_1、IgG_2、IgG_3、IgG_4和IgG浓度分别为(7.53±0.14)g/L、3.99(3.13,5.02)g/L、0.49(0.30,0.70)g/L、0.53(0.26,0.93)g/L、12.2(10.5,14.1)g/L;血清IgG_1/IgG、IgG_2/IgG、IgG_3/IgG和IgG_4/IgG分别为(61.3±0.69)%、33.38(27.8,38.8)%、3.97(2.5,5.3)%和4.44(2.1,7.3)%。女性血清IgG_3浓度及IgG_3/IgG比值高于男性(P=0.005,0.014);不同性别间IgG_1、IgG_2、IgG_4及IgG_1/IgG、IgG_2/IgG、IgG_4/IgG的差异无统计学意义。31~40岁组血清IgG_3浓度显著高于41~50岁组(P=0.03),而年龄对血清IgG_1、IgG_2和IgG_4浓度的影响无统计学意义。重度吸烟组血清IgG_1的浓度比不吸烟组低,差异有统计学意义(P=0.023)。重度吸烟组IgG_4/IgG高于不吸烟组(P=0.018)。中/重度饮酒组血IgG_1、IgG_3浓度和IgG_3/IgG比值比不饮酒组低(P=0.05,0.004,0.015)。代谢综合征低风险组血清IgG_3和IgG_3/IgG高于高风险组(P=0.034,0.038)。结论性别和年龄对于血清IgG_3浓度的影响有显著意义。重度吸烟可能导致IgG_1的浓度降低和IgG_4/IgG比值的上升。血清IgG_1、IgG_3和IgG_3/IgG的降低与饮酒存在一定的关系。     

The SAMe-TT2R2 score and decision-making between a vitamin K antagonist or a non-vitamin K antagonist oral anticoagulant in patients with atrial fibrillation

Oral anticoagulation therapy is essential in patients with atrial fibrillation and clinicians need guidance on decision-making between the vitamin K antagonists (VKA), e.g. warfarin, or non-vitamin K antagonist oral anticoagulants. Observational studies have shown that patients who receive VKA therapy spend a significant percentage of their time with international normalized ratio values outside of the therapeutic range (time in therapeutic range, TTR <60%.) Recently, a clinical score has been developed with commonly encountered clinical features, the SAMe-TT₂R₂ score, to help decision-making with regard to whether a patient is likely to do well, or not, with a VKA. Those with a SAMe-TT₂R₂ score of 0–1 are likely to do well on a VKA, while those with a SAMe-TT₂R₂ score ≥2 are on probability going to achieve suboptimal TTR. In this article, we provide an overview of the main published retrospective and prospective studies that have validated the SAMe-TT₂R₂ score and its value for decision-making in daily clinical practice.     

Selective monitoring of vitamin D2 and D3 supplementation with a highly specific 25-hydroxyvitamin D3 immunoassay with negligible cross-reactivity to 25-hydroxyvitamin D2

Background

The effects of vitamin D₂ and D₃ supplementation on circulating concentrations of 25(OH)D₃ require reliable analytical tools for specific determination of 25(OH)D₃ and 25(OH)D₂. We have developed a highly specific 25-OH Vitamin D3 ELISA with negligible cross-reactivity towards 25(OH)D₂.

Methods

25(OH)D₃ concentrations were measured in several study participants; 1) 641 healthy men and women; 2) 39 postmenopausal women receiving 400-800 IU vitamin D₃ daily for 4 months; 3) 45 men and women with hip fracture receiving 1000 IU vitamin D₂ daily for 3 months.

Results

This 25-OH Vitamin D3 ELISA had minimal cross-reactivity to 25(OH)D₂, (0.7%), and demonstrated a high correlation (r² = 0.93) with 25(OH)D₃ determined by HPLC. 25(OH)D₃ increased by 14% in subjects receiving vitamin D₃ for 4 months (p < 0.01), whereas there was no significant change in 25(OH)D₃ levels in those receiving vitamin D₂.

Conclusions

We report that 25(OH)D₃ ELISA was used for evaluation of 25(OH)D₃ concentrations in subjects receiving vitamin D₂ and D₃ supplementation. The increase of 25(OH)D₃ in circulation with vitamin D3 supplementation and lack of increase with vitamin D₂ supplementation suggest that this assay has sufficient sensitivity and specificity to be used as a reliable measurement of nutritional vitamin D₃ status in humans.     

Circadian Variation in Circulating Platelet Aggregates

The mitochondrial F₁F_o adenosine triphosphate (ATP) synthase is one of the most thoroughly studied enzyme complexes known. Yet, a number of new observations suggesting that the enzyme is also located on the cell surface necessitate further investigation. While the mitochondrial synthase utilizes the proton gradient generated by oxidative phosphorylation to power ATP synthesis, the cell surface synthase has instead been implicated in numerous activities, including the mediation of intracellular pH, cellular response to antiangiogenic agents, and cholesterol homeostasis. Intriguingly, a common thread uniting these various models of cell surface ATP synthase functions is the apparently caveolar distribution of the enzyme. Recent studies concerning the cell surface ATP synthase manifest applications in the regulation of serum cholesterol levels, cellular proliferation and antitumor strategies. This review addresses the expression, interactions, functions, and consequences of inhibition of cell surface ATP synthase, an enzyme now displaying a shift in paradigm, as well as of location.     

Vitamin B12 deficiency: New data on an old disease

Cobalamin deficiency is a common finding. In the elderly the prevalence is 10-20%, but only 5-10% of these are clinically symptomatic. Typical clinical symptoms include macrocytic anemia, neuropsychiatric symptoms and glossitis. In many cases this triad is lacking, however. The serum cobalamin assay is the best first line test, but the results must be carefully interpreted, since a normal level does not exclude deficiency. Markers of cobalamin activity, such as serum homocysteine or methylmalonic acid may be helpful in this situation. The main cause of cobalamin deficiency is atrophic gastritis. It is either caused by an autoimmune process which leads to achlorhydria and severe intrinsic factor deficiency ("classical pernicious anemia") or by atrophic gastritis from other causes, in particular helicobacter pylori infection. In the latter cases the lack of gastric acid does not allow separation of cobalamin from proteins, but intrinsic factor, although low, is sufficient for cobalamin protection (food cobalamin malabsorption). Helicobacter pylori eradication may cure some of these patients. While in food cobalamin malabsorption syndrome small doses of oral cobalamin are effective, parenteral therapy or high oral doses are required for treatment of pernicious anemia. While almost all patients respond hematologically, only half of the patients with neurological signs, and a small minority of psychiatric patients respond to treatment. Patients with pernicious anemia and atrophic gastritis have a greatly increased long-term risk for gastric carcinoids.     

Conjunctival oxygen tension monitoring in emergency department patients

Conjunctival oxygen tension (PcjO2) was sequentially monitored in 96 medical and surgical patients admitted to the emergency department resuscitation suite during a 6-month period. There were 28 patients with cardiac arrest, 44 with major trauma, and 24 with severe medical problems. A total of 2,392 PcjO2 data points were collected in these patients. In patients with cardiac arrest, PcjO2 showed changes in physiological condition as early as or earlier than measurement of vital signs. Measurement of PcjO2 and the finding of a PcjO2 index (PcjO2/PaO2) less than .5 in normotensive multiple trauma patients allowed rapid detection of hemorrhagic hypovolemia. In critically ill medical patients, low values for PcjO2 were found with hypoxemia as well as in conditions associated with decreased cardiac output and tissue oxygen delivery. These two conditions could be distinguished by measuring PaO2 and calculating the PcjO2 index; a PcjO2 index less than .5 was associated with diminished peripheral perfusion and cardiac output, and a PcjO2 index greater than .5 indicated hypoxemia without any compromise in cardiac output. In the group of critically ill surgical and medical patients included in this study, conjunctival oxygen monitoring provided clinically useful information not available from vital signs and permitted identification of physiological instability associated with abnormalities in peripheral tissue perfusion and oxygenation as early as or earlier than conventional monitoring methods.     

Reliability of Cardiac Output Calculation by the Fick Principle and Central Venous Oxygen Saturation in Emergency Conditions

Background  For many years thermodilution has been the gold standard for determining cardiac output in the critically ill patients. Less invasive methods have recently been introduced. This study aimed at evaluating the agreement between cardiac output (CO) measured by a new Fick method, using central venous saturation (Scvo₂), and that measured by the classic thermodilution technique, in patients requiring emergent CO evaluation. Settings  Prospective clinical study in a university-affiliated, tertiary hospital, at surgical and general intensive care units. Patients and methods  Fifteen mechanically ventilated patients arriving in the emergency department in hemodynamic shock, had immediately a pulmonary artery catheter introduced under fluoroscopy upon arrival into the ICU. Cardiac output (CO) was obtained in each patient via both thermodilution and the Fick method, using oxygen consumption, SpO₂ and Scvo₂. Results  COs ranged between 2 and 2.3 (in the Fick and thermodilution methods, respectively) and 19 or 19.5 l/min (respectively). Mean thermodilution-derived CO was 6.2 ± 4.2 l/min whereas the Fick’s was 7.0 ± 4.3 l/min. There was statistical significant correlation between the two modalities of measurements, with an r ² = 0.9 (P < 0.001). Conclusions  The new method of Fick assessed emergent CO as reliably as the thermodilution, regardless of whether it was low or high. The use of Scvo₂ allows for prompt bedside calculation for most emergency patients. Avi A. Weinbroum and Philippe Biderman concurred equally to the present investigation. Weinbroum AA, Biderman P, Soffer D, Klausner JM, Szold O. Reliability of cardiac output calculation by the Fick principle and central venous oxygen saturation in emergency conditions.     

Human chorionic gonadotropin (hCG) may be a marker of systemic oxidative stress in normotensive and preeclamptic term pregnancies

Objectives:

In vitro studies on placental function have revealed interactions between levels of secretion of human chorionic gonadotropin (hCG) by trophoblastic cells and oxidative stress generated by hydrogen peroxide (H₂O₂). Here, we have examined the relationship between maternal levels of hCG and H₂O₂ in vivo in term pregnancies with and without preeclampsia.

Design and methods:

We measured serum levels of hCG and H₂O₂ in twenty preeclamptic and twenty normotensive term pregnant women (controls), using an enzymatic immunoassay and an electrochemical method, respectively.

Results:

Higher levels of serum hCG and H₂O₂ were observed in patients with preeclampsia in comparison to controls. A significant positive correlation between serum hCG concentration and H₂O₂ production was found.

Conclusion:

Our results show that: (1) systemic hCG levels are correlated with an oxidative stress state in term pregnant women with preeclampsia and (2) circulating hCG may be a monitoring tool of oxidative stress during pregnancy.     

Elimination of 14C-glycated albumin from serum of rabbit

Glycated albumin was prepared by incubation of serum from rabbits with randomly labelled ¹⁴C-glucose. The isolated glycated albumin fraction was re-infused to the same animal. ¹⁴C-labelled glucose was given to alloxan-treated rabbits. The disappearance of the radioactivity showed a rapid initial phase and a slow elimination phase, which could be acceptably described by first-order kinetics. The estimated half-life of glycated albumin was about 6 days, which is about 70% of that generally stated for albumin in the rabbit. If these findings were transferred to human conditions, the half-life of fructosamine would be in the range of 13-14 days.