Triphenyl phosphate-induced pericardial edema is associated with elevated epidermal ionocytes within zebrafish embryos

Triphenyl phosphate (TPHP) is an organophosphate ester-based plasticizer and flame retardant. The objective of this study was to identify the potential role of epidermal ionocytes in mediating TPHP-induced pericardial edema within zebrafish embryos. Exposure to TPHP from 24 to 72 h post fertilization (hpf) resulted in a significant increase in pericardial edema and the number of ionocytes at 72 hpf relative to time-matched embryos treated with vehicle. In addition, co-exposure of embryos to mannitol (an osmotic diuretic) blocked TPHP-induced pericardial edema and effects on ionocyte abundance. However, knockdown of ATPase1a1.4 – an abundant Na⁺/K⁺-ATPase localized to epidermal ionocytes – mitigated TPHP-induced effects on ionocyte abundance but not pericardial edema, whereas co-exposure of embryos to ouabain – a Na⁺/K⁺-ATPase inhibitor – enhanced TPHP-induced pericardial edema but not ionocyte abundance. Overall, our findings suggest that TPHP may have multiple mechanisms of toxicity leading to an increase in ionocyte abundance and pericardial edema within developing zebrafish embryos.     

Comparison of the protective effects of curcumin and caffeic acid phenethyl ester against doxorubicin-induced testicular toxicity

Whether testicular toxicity is mediated by matrix metalloproteinases (MMPs) is an important question that has not been examined. This study investigated the suppressive effect of curcumin and caffeic acid phenethyl ester (CAPE) on oxidative stress, apoptosis, and whether MMPs mediate doxorubicin (DOX)-induced testicular injury. Male rats were randomly divided into eight groups (n = 8 per group). The groups were as follows: sham, dimethyl sulphoxide (100 µL), DOX (3 mg/kg), CAPE (2.68 mg/kg), curcumin (30 mg/kg), DOX+CAPE (3 mg/kg DOX and 2.68 mg/kg CAPE), DOX+curcumin (3 mg/kg DOX and 30 mg/kg curcumin) and DOX+CAPE+curcumin (3 mg/kg DOX, 2.68 mg/kg CAPE and 30 mg/kg curcumin). Injections were administered daily for 21 days. The oxidative stress, MMPs, proinflammatory cytokines and apoptotic markers in the DOX group were higher than the sham group (p < .05); these measures were lower in the groups treated with CAPE and curcumin together with DOX compared with the DOX group (p < .05). The results showed that MMPs mediated DOX-induced testicular injury, but CAPE and especially curcumin suppressed testis injury and cell apoptosis by suppressing DOX-induced increases in MMPs, oxidative stress and proinflammatory cytokines. However, curcumin exhibited more pronounced effects than CAPE in terms of all studied parameters.     

Assessment of neurotoxic effects of tri-cresyl phosphates (TCPs) and cresyl saligenin phosphate (CBDP) using a combination of in vitro techniques

Environmental exposures to tri-cresyl phosphates (TCPs) and the possible formation of toxic metabolites (e.g. cresyl saligenin phosphate; CBDP) may cause a variety of neurotoxic effects in humans. As reported for other organophosphorus compounds (OPs), the inhibition of acetylcholine esterase (AChE) has also been proposed as the underlying mechanism for TCP neurotoxicity. The ortho-isomer, ToCP and its metabolite CBDP are also known to affect neuropathy target esterase (NTE) leading to organophosphate-induced delayed neuropathy (OPIDN). Recently, in vitro testing has led to the identification of other molecular targets and alternative mechanisms of ToCP toxicity. The metabolite CBDP and other isomers, as well as commercial mixtures have not been tested for such additional modes of actions. Accordingly, the present study investigates alterations of neurobiological correlates of central nervous processes using different in vitro techniques. The three symmetric TCP isomers – ToCP, TpCP, and TmCP – that contain a methyl group at the ortho-, para-, or meta-position of the aromatic ring system, respectively, together with a commercial TCP mixture, and CBDP were all tested using concentrations not exceeding their cytotoxic concentrations. Isolated cortical neurons were kept in culture for 6 days followed by 24 h incubation with different concentrations of the test compounds. Thus, all endpoints were assessed after 7 days in vitro (DIV 7), at which time cell viability, neurite microstructure, and the function of glutamate receptors and voltage-gated calcium cannels (VGCC) were measured. While the cytotoxic potential of the TCP isomers and their mixture were comparable (IC₅₀ ≥ 80 μM), CBDP was more cytotoxic (IC₅₀: 15 μM) to primary cortical neurons. In contrast, CBDP (up to 10 μM) did not compromise the microstructure of neurites. Ten μM of ToCP significantly reduced the size and complexity of neurite networks, but neither TmCP and TpCP nor the mixture affected this second endpoint of neurotoxicity assessment. TCPs and their mixture significantly reduced the Ca²⁺ influx in response to glutamate and KCl stimulation in concentrations of 10 μM. Only ToCP showed a specific effect on glutamate receptors with 100 nM reducing the evoked Ca²⁺ influx. The effects of CBDP on the provoked Ca²⁺ influx were much weaker than those observed for TCPs. These results confirmed that ToCP has a unique mode of action on glutamate receptors that are not observed with the metabolite CBDP and the other symmetric TCP isomers. In addition, the TmCP isomer seems to have the lowest potency with respect to inducing neurotoxic effects. CBDP did not affect the neurospecific endpoints investigated in this study. Therefore, the specific affinity of CBDP for NTE and the reported general cytotoxicity might be the most relevant modes of action of this toxic metabolite in the context of ToCP-induced neurotoxicity, including OPIDN.     

Contribution of neutrophil elastase to the lysis of obliterative thrombi in the context of their platelet and fibrin content

Leukocytes invade newly formed thrombi through interactions with platelets and fibrin and later contribute to the removal of fibrin deposits mainly through the action of neutrophil elastase. The present study attempts to express in quantitative terms the impact of neutrophils on the lytic processes in obliterative thrombi based on the local presence of elastase-specific fibrin degradation products (NE-FDP) in relation to the leukocyte, platelet and fibrin content of thrombi. Immunofluorescent detection of fibrin, NE-FDP and platelet antigens was performed in sections of thrombi from 28 patients subjected to thrombectomy in combination with DNA-staining for identification of nucleated cells. The digitalized fluorescent microscopic images were decomposed according to the color channel of each thrombus constituent. The integrated intensity values for all thrombus constituents were statistically evaluated with correlation, hierarchical agglomerative clustering , Hotelling's T² and F-statistics. Association between NE-FDP and leukocyte content of thrombi is evidenced by a significant Pearson correlation coefficient of 0.71 (p = 0.00002). Cluster analysis reveals two classes of thrombi according to NE-FDP, leukocyte and platelet content and also two according to NE-FDP, leukocyte and fibrin content. When NE-FDP, fibrin and platelet content is normalized to the leukocyte count in the same thrombus, clusters with platelet-related thrombolytic resistance (inversely related NE-FDP and platelet content) and advanced cell-dependent thrombolysis (inversely related NE-FDP and fibrin content) are identified. These distinct patterns of thrombus constituents are snapshots of characteristic stages in the cell-dependent thrombolysis, which indicate a clot-stabilizing role for platelets in this process similar to their impact on the plasmin-dependent lysis.     

工业大麻地上部分化学成分研究

目的 研究工业大麻Cannabis sativa地上部分的化学成分.方法 采用硅胶柱色谱及HPLC等色谱技术进行分离纯化,通过理化性质与波谱数据分析鉴定结构.结果 从工业大麻地上部分的醋酸乙酯萃取物中分离得到30个单体化合物,分别为13S-羟基-9Z,11E,15E-十八碳三烯酸甲酯(1)、△9-四氢次大麻酚酸(2)、...     

Uptake and effects of manufactured silver nanoparticles in rainbow trout (Oncorhynchus mykiss) gill cells

Nanoparticles are already widely used in technology, medicine and consumer products, but there are limited data on their effects on the aquatic environment. In this study the uptake and effect of citrate (AgNP_CIT) and polyvinylpyrrolidone (AgNP_PVP) coated manufactured silver nanoparticles, as well as AgNO₃ (Ag⁺) were tested using primary gill cells of rainbow trout (Oncorhynchus mykiss). Prior to use, the nanoparticles were characterized for size, surface charge and aggregation behavior. Gill cells were cultured either as monolayers on solid support, or as multilayers on a permeable support cell culturing system, enabling transport studies. The uptake of silver nanoparticles and Ag⁺ after exposure to 10 mg L⁻¹ was determined with microscopical methods and inductively coupled plasma mass spectrometry (ICP-MS). Cytotoxicity, in terms of membrane integrity, as well as oxidative stress (depletion of reduced glutathione) was tested at silver concentrations ranging from 0.1 mg L⁻¹ to 10 mg L⁻¹. Results show that AgNP_CIT nanoparticles are readily taken up into gill cell monolayers while uptake was less for AgNP_PVP. In contrast, it appears that the slightly smaller AgNP_PVP were transported through cultured multilayers to a higher extent, with transport rates generally being in the ng cm⁻² range for 48 h exposures. Transport rates for all exposures were dependent on the epithelial tightness. Moderate cytotoxic effects were seen for all silver treatments. Levels of reduced glutathione were elevated in contrast to control groups, pointing on a possible overcompensation reaction. Taken together silver nanoparticles were taken up into cells and did cause silver transport over cultured epithelial layers with uptake and transport rates being different for the two nanoparticle species. All silver treatments had measurable effects on cell viability.     

大剂量维生素C联合磷酸肌酸钠治疗小儿病毒性心肌炎的临床研究

目的 探讨大剂量维生素C联合磷酸肌酸钠对于病毒性心肌炎小儿患者的治疗效果.方法 选取在我科治疗的小儿病毒性心肌炎120例,随机分为对照组和观察组各60例,对照组采取常规抗病毒,吸氧等对症处理,观察组在此基础上给予大剂最维生素C联合磷酸肌酸钠治疗,比较两组患者的临床治疗效果及心肌酶谱的变化.结果 ①经比较,观察组CK、CK-MB、HBDH恢复正常时间均明显短于对照组(P＜0.05).②观察组心电图以及临床症状体征改善明显优于对照组(P＜0.05).结论 大剂量维生素C联合磷酸肌酸钠对心肌损害具有明显的保护作用,临床使用能够减轻病情,缩短疗程,增强疗效.     

正交设计法优选草乌炮制工艺

用正交设计,以草乌炮制前后总生物碱和酯型碱含量变化为指标,研究草乌炮制工艺。试验数据经用多指标试验公式评分法处理,选择出草乌的最佳炮制工艺为水浸润透,切厚片,加压(127℃,0.15MPa)蒸3h。     

墓头回环烯醚萜酯提取部位抗肿瘤作用及机制研究

目的 研究墓头回醋酸乙酯提取物环烯醚萜酯部位（PHEBB）的体内外抗肿瘤作用及其机制。方法 硅胶柱色谱法精制PHEBB;反相高效液相色谱法分析PHEBB化学成分。MTT法检测PHEBB对人肿瘤细胞KB、COLO-205、SGC-7901的抑制活性。采用小鼠移植性瘤模型研究PHEBB对小鼠S₁₈₀肉瘤、H₂₂肝癌的体内抗肿瘤作用;免疫组化法研究PHEBB对肿瘤组织中相关基因表达的影响。结果 PHEBB主要成分均为环烯醚萜酯类,主要有desacetylisovaltratum、isovaltrate acetoxyhydrin。体外实验结果显示,PHEBB对KB、COLO-205、SGC-7901等癌细胞均具有显著抑制作用,IC₅₀在2.27～5.49 μg/mL。体内实验显示,PHEBB以60 mg/kg ip给予荷瘤小鼠,对S₁₈₀肉瘤的抑制率为45.5%,对H₂₂肝癌的抑制率为54.2%;免疫组化检测结果显示,PHEBB能上调小鼠H₂₂肿瘤组织中Bax表达,下调Bcl-2表达,且能降低肿瘤组织中微血管密度。结论 PHEBB抗肿瘤药效物质为环烯醚萜酯;其体内外均具有一定抗肿瘤作用,且其抗肿瘤机制可能与诱导肿瘤细胞凋亡及抑制肿瘤微血管有关。