P(HPMA-FMA)荧光探针制备、细胞毒性评估及细胞吞噬示踪检测

以甲基丙烯酰氯、三乙胺和荧光素反应得到荧光素甲基丙烯酸酯(FMA),将其与聚N-(2-羟丙基)甲基丙烯酰胺(HPMA)以物质的量之比1∶10混合并通过引发剂偶氮二异丁腈(AIBN)引发聚合反应,生成带有荧光探针的聚合物P(HPMA-FMA)。采用台盼蓝排染法评估了该聚合物的细胞毒性,荧光显微镜和流式细胞仪观察和检测了全反式维甲酸(ATRA)诱导HL-60细胞分化过程中,P(HPMA-FMA)被细胞吞噬后的荧光示踪效应。结果表明:P(HPMA-FMA)的细胞毒性极低,当P(HPMA-FMA)的质量浓度为4~16mg/mL时对细胞增殖无影响;当其质量浓度为30μg/mL时即可满足荧光显微镜定性示踪观察和流式细胞术定量检测所需要的荧光强度。     

锌对鲫鱼免疫功能的影响研究

以鲫鱼为实验对象,探讨了饲料中锌质量分数对鱼类免疫功能的影响.通过对鱼体免疫组织生物发光,抗体的凝集效价,腹腔巨噬细胞(Macrophage,Mφ)吞噬活性等免疫指标进行研究,结果发现饲料中锌质量分数与鲫鱼的免疫机能密切相关,缺锌时鲫鱼生物发光值,抗体的凝集效价均为最低,Mφ吞噬活性最弱,但高锌同样也会抑制其免疫功能.说明饲料中锌的质量分数能影响鲫鱼的免疫功能,一般需要量为38～80 mg/kg饲料,不宜过高.     

Arginine Deiminase Induces Immunogenic Cell Death and Is Enhanced by N-acetylcysteine in Murine MC38 Colorectal Cancer Cells and MDA-MB-231 Human Breast Cancer Cells In Vitro

The use of arginine deiminase (ADI) for arginine depletion therapy is an attractive anticancer approach. Combination strategies are needed to overcome the resistance of severe types of cancer cells to this monotherapy. In the current study, we report, for the first time, that the antioxidant N-acetylcysteine (NAC), which has been used in therapeutic practices for several decades, is a potent enhancer for targeted therapy that utilizes arginine deiminase. We demonstrated that pegylated arginine deiminase (ADI-PEG 20) induces apoptosis and G0/G1 phase arrest in murine MC38 colorectal cancer cells; ADI-PEG 20 induces Ca²⁺ overload and decreases the mitochondrial membrane potential in MC38 cells. ADI-PEG 20 induced the most important immunogenic cell death (ICD)-associated feature: cell surface exposure of calreticulin (CRT). The antioxidant NAC enhanced the antitumor activity of ADI-PEG 20 and strengthened its ICD-associated features including the secretion of high mobility group box 1 (HMGB1) and adenosine triphosphate (ATP). In addition, these regimens resulted in phagocytosis of treated MC38 cancer cells by bone marrow-derived dendritic cells (BMDCs). In conclusion, we describe, for the first time, that NAC in combination with ADI-PEG 20 not only possesses unique cytotoxic anticancer properties but also triggers the hallmarks of immunogenic cell death. Hence, ADI-PEG 20 in combination with NAC may represent a promising approach to treat ADI-sensitive tumors while preventing relapse and metastasis.     

Immunotoxicity of butyltins in tunicates

This study was designed to investigate the effects of butyltins on yeast phagocytosis in vitro by haemocytes of the colonial ascidian Botryllus schlosseri. This species has been reported to be very sensitive to organotins. Results show that, in analogy to reports on mammalian leukocyte re-activity, butyltins exert inhibitory effects on phagocytosis in a concentration-dependent manner, mainly by influencing cellular calcium homeostasis by interacting with the calcium pump. As the immunosuppressant activity of organotins described in mammals and teleosteans also occurs in an invertebrate marine species, the latter may assume a role as a sensitive biosensor of butyltins as immunotoxins in aqueous ecosystems.     

In vitro effects of methylmercury on ascidian (Styela plicata) immunocyte responses

This study shows that high methylmercury concentrations are cytotoxic for Styela plicata hemocytes, whereas sublethal concentrations affect immunocyte responses. Moreover, hemocytes exposed to the xenobiotic present a significantly enhanced phenoloxidase activity as revealed in the hemocyte lysate supernatant compared with the control. Although the cytotoxic activity of S. plicata hemocytes toward rabbit erythrocytes is a PO‐dependent cell‐target reaction due to quinone products, it was significantly decreased by suitable methylmercury concentrations in the medium. The same xenobiotic concentrations decreased the hemocyte phagocytic activity toward yeast. In both the responses cell‐target contacts could be affected by methylmercury, whereas the releasing capacity appeared to be unchanged, as indicated by hemocyte PO‐release in the medium. Finally, changes in hemocyte shape and spreading capacity were shown. On the basis of the present results, Styela plicata hemocyte responses could be an additional immunotoxicology test using a microplate method that reveals cell morphological changes and spreading capacity. Copyright © 2007 John Wiley & Sons, Ltd.     

Imaging Macrophage Phagocytosis Using AIE Luminogen‐Labeled E. coli

Phagocytosis of bacteria is an important biological process. Gaining insight into this process may greatly benefit related pathological studies and further contribute to development of therapies for infectious diseases. Tools for studying these internalization processes, however, are limited. Herein, we demonstrate the feasibility of employing an environmentally sensitive aggregation‐induced emission (AIE) probe for bacteria labeling and imaging. By tracking the fluorescence variation of the stained bacteria, the pH changes of its microenvironment can be monitored. In this way, the phagocytic entry of these bacteria into the macrophage cells and the intravacuolar acidification can be visualized in real‐time.     

乙肝疫苗注入后婴儿血液吞噬细胞吞噬功能变化的临床观察

为探讨乙开型肝炎表面抗原（ｈｅｐａｔｉｔｉｓ Ｂ ｓｕｒｆａｃｅ ａｎｔｉｇｅｎ,ＨＢｓＡｇ）阳性母亲的婴儿乙肝疫苗注入后早期机体免疫反应的特点,采用化我法测定了吞噬细胞的吞噬功能;用双抗原夹心法测定了抗－ＨＢｓ抗体。结果表明,（１）乙肝疫苗注入前两组间吞噬细胞的吞噬功能无明显差异,疫苗注入后３个月和６个月时实组外周血吞噬细胞的吞噬功能明显下降（Ｐ〈０．０５）。两组间同时间点相比,１个月和３个月都     

Exploring the Biocompatibility of Zwitterionic Copolymers for Controlling Macrophage Phagocytosis of Bacteria

This paper provides a biomaterial derived from zwitterionic polymer for controlling macrophage phagocytosis of bacteria. A series of zwitterionic copolymers, named DMAPS‐co‐AA, are synthesized with 3‐dimethyl (methacryloyloxyethyl) ammonium propane sulfonate (DMAPS) and acrylic acid (AA). The biocompatibility of DMAPS‐co‐AA copolymers can be adjusted by adjusting the DMAPS‐content or pH value. As the DMAPS‐content increases, the biocompatibility of zwitterionic copolymer increases. The zwitterionic copolymers with DMAPS content above 30 wt% have higher biocompatibility. Moreover, the biocompatibility also increases significantly as the pH increases from 3.4 to 7.2. By adjusting the pH above 5.8, the zwitterionic copolymer with lower DMAPS‐content also shows higher biocompatibility. Importantly, after incubation with the DMAPS‐co‐AA copolymer solutions at different pH values, phagocytosis behavior of macrophage RAW264.7 cells can also be adjusted. The phagocytosis of bacteria is enhanced at pH = 7.2. Thus, it is proposed that zwitterionic copolymers can be used for controlling phagocytosis of bacteria.

     

A Photostable AIEgen for Specific and Real‐time Monitoring of Lysosomal Processes

Lysosomes are recognized as advanced organelles involved in many cellular processes and are also considered as crucial regulators of cell homeostasis. The current strategies for monitoring activities of lysosomes exhibit some limitations. Herein, we synthesized a novel fluorescent probe named 2M‐DPAS with AIE characteristics, which was proved to have significant advantages of good biocompatibility, high selectivity, bright emission and excellent photostability. Based on those, 2M‐DPAS can be used to continuously monitor the dynamic changes of lysosomes, including autophagy and mitophagy, as well as to track the process of endocytosis of macromolecules in lysosomes, which are of benefit to better know about the lysosomes‐related diseases.     

中华圆田螺血细胞的分类和吞噬活性

血细胞作为主要的免疫效应细胞在贝类的免疫和防御中有重要作用。本文通过相差显微镜和透射电子显微镜对中华圆田螺(Viviparus chinensis)血细胞的形态与类型以及血细胞对枯草芽孢杆菌的吞噬观察。结果发现壳高4～6 cm的田螺血细胞浓度为(2.17±0.38)×106个/mL。根据血细胞大