Marked hyperphosphatasemia associated with an acute leukemia in a Great Dane

This is the report of a 5‐year‐old male neutered Great Dane with an extreme leukocytosis (544.9 × 10⁹ cells/L; RI 5.2–13.9 × 10⁹ cells/L) characterized by highly atypical round cells. Cellular morphologic features such as cytoplasmic membrane blebs, a high nuclear‐to‐cytoplasmic ratio, and nuclear indentations and irregularities and large nucleoli, as well as immunocytochemistry for CD3 and CD79, myeloperoxidase cytochemistry, and clonality testing were not conclusive for myeloid or lymphoid origin. Marked alkaline hyperphosphatasemia was present at the first visit (2783.0 U/L; RI 6–80.0 U/L), followed by a 5‐fold increase (14,000 U/L) a week later, identified as being mostly contributed by the bone‐ALP isoform (11,062 U/L; RI 0–30 U/L). In addition, the atypical leukocytes were strongly positive for cytoplasmic ALP activity. In vitro lysis of a heparin blood sample resulted in a 1.7‐fold increase of ALP activity, supporting the origin of the hyperphosphatasemia at least in part from the leukemic cell population. To the authors’ knowledge, this is a unique case of alkaline hyperphosphatasemia, due at least to a leukemic cell population producing a bone‐ALP isoform, regardless of the exact nature of the leukemia.     

Semi-Synthesis,Cytotoxic Evaluation,and Structure—Activity Relationships of Brefeldin A Derivatives with Antileukemia Activity

Brefeldin A (1), a potent cytotoxic natural macrolactone, was produced by the marine fungus Penicillium sp. (HS-N-29) from the medicinal mangrove Acanthus ilicifolius. Series of its ester derivatives 2–16 were designed and semi-synthesized, and their structures were characterized by spectroscopic methods. Their cytotoxic activities were evaluated against human chronic myelogenous leukemia K562 cell line in vitro, and the preliminary structure–activity relationships revealed that the hydroxy group played an important role. Moreover, the monoester derivatives exhibited stronger cytotoxic activity than the diester derivatives. Among them, brefeldin A 7-O-2-chloro-4,5-difluorobenzoate (7) exhibited the strongest inhibitory effect on the proliferation of K562 cells with an IC₅₀ value of 0.84 µM. Further evaluations indicated that 7 induced cell cycle arrest, stimulated cell apoptosis, inhibited phosphorylation of BCR-ABL, and thereby inactivated its downstream AKT signaling pathway. The expression of downstream signaling molecules in the AKT pathway, including mTOR and p70S6K, was also attenuated after 7-treatment in a dose-dependent manner. Furthermore, molecular modeling of 7 docked into 1 binding site of an ARF1–GDP-GEF complex represented well-tolerance. Taken together, 7 had the potential to be served as an effective antileukemia agent or lead compound for further exploration.     

Recording cerebellar neuron activities in swimming goldfish

ABSTRACT:   Neuronal activities were investigated in the cerebellum of immobilized and swimming goldfish Carassius auratus . Extracellularly recorded neural activities of the cerebellum in immobilized goldfish were characterized and classified into five types. Based on the waveforms and recording depths, these five neural activity types were estimated to originate from three identified classes of cerebellar neurons: Purkinje cells, eurydendroid cells, and granule cells. Chronic recording of cerebellar neuron activities in unrestrained goldfish was performed for more than 100 h. During the chronic recordings, a submersible amplifier attached to the goldfish head, and a multielectrode array developed for the present study were used. Neuronal activities in the cerebellum of free-swimming fish could also be classified into five types as in the immobilized condition. Firing patterns of two neurons identified as Purkinje cells and eurydendroid cells were analyzed during turning movements of the goldfish. The firing patterns of these neurons changed in relation to turning movements. Although some improvements are required, the chronic recording method developed in the present study can be applied to further investigations concerning the direct relationship between brain neural activities and certain behavior.     

A chronic oral toxicity study of marine collagen peptides preparation from chum salmon (Oncorhynchus keta) skin using Sprague-Dawley rat

Due to the increased consumption of marine collagen peptides preparation (MCP) as ingredients in functional foods and pharmaceuticals, it was necessary to carry out safety requirements in the form of an oral chronic toxicity assessment. In order to define the oral chronic toxicity of MCP, a 24-month feeding study of MCP was carried out. Sprague-Dawley (S-D) rats at the age of four-week of both sexes were treated with MCP at the diet concentrations of 0%, 2.25%, 4.5%, 9% and 18% (wt/wt). The actual food intake and bodyweight of the individual animals were recorded periodically until sacrifice. Blood and urine samples were collected for serum chemistry evaluations and urinalysis. Throughout the experimental period, there was no toxicologically significant difference between the vehicle and MCP-treated animals with respect to the survival rate, body weight, food consumption, urinalysis, clinical biochemistry parameter and relative organ weight in either sex. Moreover, incidences of non-neoplastic lesions in MCP-treated groups did not significantly increase compared with the control group. Under the present experimental conditions, no higher risk of chronic toxic effects was observed in MCP-treated rats at the diet concentrations of 2.25%, 4.5%, 9% and 18% (wt/wt) than in the rats fed with basal rodent diet.     

以鲥为瘟:古代长江鲥鱼的污名化

鲥鱼在明清一些著作中被称为瘟鱼。历来关于瘟鱼的解释主要围绕鲥鱼本身来阐发,但这些观点不能很好地解释鲥鱼为何会在江西、两湖以及四川一带被称作瘟鱼,而在盛产鲥鱼的江东却没有这种指称。明人的“瘟鱼”观点源自唐人孟诜“发疳、痼”的说法,李时珍所说的蜀地瘟鱼并不是鲥鱼。一些清人认为瘟鱼并非来自鲥鱼本身。《见闻录》中记载了一个鲥鱼被虚构成瘟鱼的故事,揭示出瘟鱼是后人将瘟疫附会到鲥鱼身上的结果。显然,鲥鱼在特定地点被称为瘟鱼,是鲥鱼与地方文化结合的产物。     

钴圃慢照射小麦不同发育阶段植株的生物学效应

利用钴圃分别慢照射普通小麦减数分裂期前、配子体时期、合子幼胚期的植株。结果表明:照射合子幼胚期对当代(M_1)结实率的影响较小,对下一代(M_2)出苗率的影响较大,M_2代的出苗率可作为选择照射合子幼胚期适宜剂量的主要指标。慢照射当代是辐射损伤的主要世代,M_2代的损伤已很轻,是突变显现的第1个世代。文中讨论了照射植株的世代划分问题。     

饮水高钠致肉鸡急性和慢性肺动脉高压综合征

为了探讨肉鸡肺动脉高压综合征 (PHS)的发病机制 ,将 2 4 0羽 1日龄健康AA肉鸡随机均分为对照组 (C)、试验Ⅰ组 (T1)和试验Ⅱ组 (T2 )。从 8日龄起实验组在饮水中添加Na+ ,使水中Na+ 含量分别达 6 0 0和 12 0 0mg·L-1,动态观察和检测各组肉鸡临床表现、PHS发病率、右心与全心重量比 (RV/TV)、红细胞压积 (PCV)、红细胞滤过指数 (FI)和血容量 (BV) ,对各组患病鸡与对照鸡的肺小动脉结构变化进行动态检测和比较分析。结果显示 ,饮水高钠成功诱发了肉鸡PHS ,并显著提高了肉鸡PHS发病率、RV/TV和PCV (P <0 0 5 )。在 2 8日龄以前 ,试验组患病鸡的FI和BV显著增加 (P <0 0 5 ) ,而血管肌化百分率 (%MA和 %WTPV)、管壁面积与管总面积之比 (WA/TA)、平均中膜厚度 (mMTPA)和血管非肌化百分率 (%NMA)无显著差异。 35和 4 2日龄时 ,试验组患病鸡肺血管 %MA、%WTPV、WA/TA和mMTPA显著增加 (P <0 0 5 ) ,%NMA则显著降低 (P <0 0 5 ) ,但FI和BV无变化。从而揭示了饮水高钠使 14～ 2 1日龄的PHS患鸡出现了以肺动脉血流量增加和肺血管阻力升高的血液流变力学变化为主要特征的急性肺动脉高压综合征。使 35～ 4 2日龄的PHS患鸡出现了以肺小动脉肌型化和管壁增生肥厚的血管重构为主要特征的慢性肺动脉高压综合征。     

Celecoxib inhibits viability,induces apoptosis and inhibits autophagy in acute myeloid leukemia cell lines HL-60 and HL-60A

AIM: To investigate the effects of celecoxib on viability, apoptosis and autophagy in acute myeloid leukemia (AML) cell lines HL-60 and HL-60A. METHODS: The HL-60 cells and HL-60A cells were cultured with various concentrations (0, 20, 40, 60, 80 and 100 μmol/L) of celecoxib. The inhibitory effect of celecoxib on the cell viability was evaluated by MTT assay. Apoptosis was analyzed by Annexin-V/PI staining. Apoptosis-related and autophagy-related proteins were determined by Western blot. RESULTS: IC₅₀ of celecoxib were 49.4 μmol/L, 32.0 μmol/L and 25.1 μmol/L for HL-60 cells treated with celecoxib for 24 h, 48 h and 72 h, respectively. For HL-60A cells, the corresponding IC₅₀ were 69.1 μmol/L, 42.5 μmol/L and 29.6 μmol/L, respectively. The results of flow cytometry analysis showed the proportions of Annexin-Ⅴ⁺ PI^-, Annexin-Ⅴ⁺ PI⁺ and Annexin-Ⅴ^-PI⁺ cells were increased in the HL-60 cells, and those of Annexin-Ⅴ⁺PI^- and Annexin-Ⅴ⁺ PI⁺ cells were increased in the HL-60A cells treated with celecoxib for 24 h. After treated with celecoxib, the induction of apoptosis was observed, the apoptosis-related proteins cleaved caspase-3 and cleaved PARP were upregulated, the autophagy-related proteins LC3 II and P62 were both increased, and mTOR, p-mTOR, 4-EBP and p-4-EBP were not changed, indicating that celecoxib inhibited autophagy in the AML cells without the mTOR pathway involvement. CONCLUSION: Celecoxib inhibits the viability of HL-60 cells and HL-60A cells in a time-and dose-dependent manner by its effects of inducing apoptosis and necrosis. Celecoxib inhibits mTOR-independent autophagy in AML cells, indicating a possible way of using celecoxib for enhancing the antitumor activity of therapeutic agents to induce cytoprotective autophagy in the AML cells.     

Identification and establishment of sorting method for isolating CD11b+ myeloid cells in human hepatic metastases from colorectal cancer

AIM: To establish a method for obtaining specific cells in solid tumor tissue by sorting of CD11b⁺ myeloid cells in hepatic metastases from colorectal cancer.METHODS: Tumor tissues were prepared into single cell suspension by mechanical method combined with enzyme digestion, and then the CD11b⁺ myeloid cells were isolated by flow cytometry. The sorted cells were identified by immunocytochemistry. The viability and morphologiy of the sorted cells were evaluated by Giemsa and Typan blue staining. The cell purity was evaluated by flow cytometry.RESULTS: Sufficient numbers of CD11b⁺ cells with high purity were isolated by sorting with flow cytometry from the single cell suspension prepared by mechanical and enzyme digestion. The purity of the cells was confirmed by statistical analysis (P<0.05). The positive rates of the cells before and after sorting were significantly different (P<0.01). The positive cells were verified by immunocytochemical method. Meanwhile, the sorted cells had complete morphology and good activity.CONCLUSION: The CD11b⁺ myeloid cells in solid tumor tissue can be isolated by flow cytometry from the machine-enzyme digestion suspension with high purity, good activity and complete morphology.