碳酸酐酶1沉默对肺癌A549细胞增殖、凋亡、侵袭和迁移的影响

目的:探讨沉默碳酸酐酶1（CA1）对人肺癌A549细胞增殖、凋亡、侵袭和迁移的影响。方法:采用脂质体转染法将CA1特异性siRNA（si-CA1组）及阴性对照（si-NC组）转染肺癌A549细胞,同时以转染空脂质体的A549细胞为空白对照组（Blank组）。采用实时荧光定量PCR（qPCR）和蛋白印迹法（Western blot）检测CA1 mRNA和蛋白表达水平,细胞计数试剂盒法（CCK-8）、流式细胞术、Transwell实验分别检测A549细胞增殖、凋亡、侵袭和迁移能力。结果:qPCR和Western blot检测结果显示,转入CA1 siRNA的A549细胞中CA1 mRNA和蛋白的表达水平均明显下调（P<0.05）;CCK-8实验结果显示,沉默CA1后si-CA1组A549细胞在24、48和72 h时光密度（OD）值明显低于si-NC组（P<0.05）;流式细胞术结果发现,si-CA1组A549细胞凋亡率明显高于si-NC组（P<0.05）;Transwell实验结果显示,si-CA1组侵袭和迁移细胞数均明显少于si-NC组（P<0.05）。与Blank组相比,si-NC组以上各指标差异均不显著（P>0.05）。结论:沉默CA1能够抑制肺癌A549细胞增殖、侵袭和迁移,并促进细胞凋亡。     

碳酸酐酶Ⅸ在核医学中的研究进展

碳酸酐酶Ⅸ（Carbonic anhydrase Ⅸ,CA Ⅸ）是碳酸酐酶家族（Carbonic anhydrases,CAs）的异构体之一,在许多肿瘤中过量表达,是一种与肿瘤相关的跨膜糖蛋白。它催化CO₂水解为碳酸氢根和氢离子,维持肿瘤细胞内的pH,酸化细胞外环境,有利于肿瘤的生长和转移。在核医学的靶向诊断和治疗中,CA Ⅸ成为一个抗肿瘤药物的靶向目标,也成为病人病情的一个预后因子。CA Ⅸ抑制剂可以作为一种诊断药物,用于肿瘤的分子影像;也可以作为一种治疗药物,阻断CA Ⅸ所调控的通路。利用⁸⁹Zr、¹¹¹In、¹²⁴I 或¹²⁵I标记靶向CA Ⅸ的单克隆抗体或其片段进行分子影像,能精确定位CA Ⅸ的分布,灵敏检测CA Ⅸ的表达情况;利用⁹⁰Y、¹³¹I、¹⁷⁷Lu或¹⁸⁶Re标记靶向CA Ⅸ的单克隆抗体,在放免疫治疗中可有效延迟肿瘤的生长。本文对CA Ⅸ及其在核医学中的研究进展进行了综述。     

二氧化碳浸渍法在蓝莓酒酿造中的应用

利用二氧化碳浸渍法酿造蓝莓酒,对比二氧化碳浸渍法和传统酿造法酿造的蓝莓酒在各理化指标方面的不同,探究该方法对蓝莓酒品质的影响。同时利用高效液相色谱法（HPLC）测定CO2不同浸渍时间对于蓝莓酒中10种有机酸酸、9种酚酸含量的影响。结果表明,蓝莓酒中含量最高的有机酸是柠檬酸,最主要的酚酸是绿原酸;二氧化碳浸渍法相比于传统酿造法能够降低蓝莓酒的酸度（P＜0.05）,但浸渍时间越长,有机酸含量越高;二氧化碳浸渍法蓝莓酒总酚含量低于传统法（P＜0.05）,但能提高总黄酮和酚酸类物质的浸出。     

Single and mixed poloxamine micelles as nanocarriers for solubilization and sustained release of ethoxzolamide for topical glaucoma therapy

Polymeric micelles of single and mixed poloxamines (Tetronic) were evaluated regarding their ability to host the antiglaucoma agent ethoxzolamide (ETOX) for topical ocular application. Three highly hydrophilic varieties of poloxamine (T908, T1107 and T1307) and a medium hydrophilic variety (T904), possessing a similar number of propylene oxide units but different contents in ethylene oxide, were chosen for the study. The critical micellar concentration and the cloud point of mixed micelles in 0.9 per cent NaCl were slightly greater than the values predicted from the additive rule, suggesting that the co-micellization is hindered. Micellar size ranged between 17 and 120 nm and it was not altered after the loading of ETOX (2.7–11.5 mg drug g^–1 poloxamine). Drug solubilization ability ranked in the order: T904 (50-fold increase in the apparent solubility) > T1107 ≅ T1307 > T908. Mixed micelles showed an intermediate capability to host ETOX but a greater physical stability, maintaining almost 100 per cent drug solubilized after 28 days. Furthermore, the different structural features of poloxamines and their combination in mixed micelles enabled the tuning of drug release profiles, sustaining the release in the 1–5 days range. These findings together with promising hen''s egg test-chorioallantoic membrane biocompatibility tests make poloxamine micelles promising nanocarriers for carbonic anhydrase inhibitors in the treatment of glaucoma.     

Preparation and evaluation of in situ gelling ophthalmic drug delivery system for methazolamide

Purpose: In this study, a thermosensitive in situ gelling vehicle was prepared to increase the precorneal resident time and the bioavailability of methazolamide (MTA). Method: Poloxamer analogs were used as the gelling agents, and the in situ gel was obtained by using a cold method. The gelation temperature, rheological properties, in vitro release as well as in vivo evaluation (the elimination of MTA in aqueous humor and intraocular-lowering effect) of the optimized formulations were investigated. Results: The optimum concentrations of poloxamer analogs for the in situ gel-forming delivery system were 21% (w/w) poloxamer 407 and 10% (w/w) poloxamer P188. This formulation was able to flow freely under nonphysiological conditions and underwent sol–gel transition in the cul-de-sac upon placement into the eye. In vitro release studies demonstrated a diffusion-controlled release of MTA from the poloxamer solutions over a period of 10 hours. In vivo evaluation indicated that the poloxamer solutions had a better ability to retain drug than MTA eyedrops did. Conclusion: These results suggested that in situ gelling ophthalmic drug delivery system may hold some promise in ocular MTA delivery.     

Carbonic anhydrase inhibitors: design of membrane-impermeant copper(II) complexes of DTPA-, DOTA-, and TETA-tailed sulfonamides targeting the tumor-associated transmembrane isoform IX

The synthesis and carbonic anhydrase (CA, EC 4.2.1.1) inhibitory activity of two series of aromatic sulfonamides and their Cu(II) derivatives, incorporating metal-complexing moieties of the DTPA, DOTA, and TETA type are reported. The new compounds were designed in such a way as to possess high affinity for Cu(II) ions, exploiting four pendant carboxylate moieties in the DTPA derivatives, as well as the cyclen/cyclam macrocyles, and three pendant acetate moieties in the DOTA and TETA derivatives. The new derivatives showed modest inhibition of the cytosolic isoform CA I (K(I) values in the range of 66-2130 nM), were better CA II inhibitors (K(I) values in the range of 21-360 nM), and excellent inhibitors of the tumor-associated isoform CA IX (K(I) values in the range of 4.1-110 nM), with selectivity ratios for the inhibition of the tumor (CA IX) over the cytosolic (CA II) isozyme in the range of 10.74-20.88 for the best derivatives. Copper complexes were more inhibitory than the corresponding ligand sulfonamides, and showed membrane impermeability, thus, having the possibility to specifically target the transmembrane CA IX that has an extracellular active site. Incorporation of radioactive copper isotopes in this type of CA inhibitor may lead to interesting diagnostic/therapeutic applications for such compounds.     

不同类别盐渍土特性对比分析

通过对氯盐渍土、硫酸盐渍土及碳酸盐渍土三种盐渍土的特性进行对比分析,发现其密度、液限与塑限、强度等特性与含水量和含盐量的关系极为密切,研究盐渍土的特性对解决其对工程造成的危害具有一定的指导意义。     

Inhibitors of Mitochondrial Human Carbonic Anhydrases VA and VB as a Therapeutic Strategy against Paclitaxel-Induced Neuropathic Pain in Mice

Neuropathy development is a major dose-limiting side effect of anticancer treatments that significantly reduces patient’s quality of life. The inadequate pharmacological approaches for neuropathic pain management warrant the identification of novel therapeutic targets. Mitochondrial dysfunctions that lead to reactive oxygen species (ROS) increase, cytosolic Ca²⁺ imbalance, and lactate acidosis are implicated in neuropathic pain pathogenesis. It has been observed that in these deregulations, a pivotal role is played by the mitochondrial carbonic anhydrases (CA) VA and VB isoforms. Hence, preclinical studies should be conducted to assess the efficacy of two novel selenides bearing benzenesulfonamide moieties, named 5b and 5d, and able to inhibit CA VA and VB against paclitaxel-induced neurotoxicity in mice. Acute treatment with 5b and 5d (30–100 mg/kg, per os – p.o.) determined a dose-dependent and long-lasting anti-hyperalgesic effect in the Cold plate test. Further, repeated daily treatment for 15 days with 100 mg/kg of both compounds (starting the first day of paclitaxel injection) significantly prevented neuropathic pain development without the onset of tolerance to the anti-hyperalgesic effect. In both experiments, acetazolamide (AAZ, 100 mg/kg, p.o.) used as the reference drug was partially active. Moreover, ex vivo analysis demonstrated the efficacy of 5b and 5d repeated treatments in reducing the maladaptive plasticity that occurs to glia cells in the lumbar portion of the spinal cord and in improving mitochondrial functions in the brain and spinal cord that were strongly impaired by paclitaxel-repeated treatment. In this regard, 5b and 5d ameliorated the metabolic activity, as observed by the increase in citrate synthase activity, and preserved an optimal mitochondrial membrane potential (ΔΨ) value, which appeared depolarized in brains from paclitaxel-treated animals. In conclusion, 5b and 5d have therapeutic and protective effects against paclitaxel-induced neuropathy without tolerance development. Moreover, 5b and 5d reduced glial cell activation and mitochondrial dysfunction in the central nervous system, being a promising candidate for the management of neuropathic pain and neurotoxicity evoked by chemotherapeutic drugs.     

New Developments in Carbonic Anhydrase IX-Targeted Fluorescence and Nuclear Imaging Agents

Carbonic anhydrase IX (CAIX) is a tumor-specific and hypoxia-induced biomarker for the molecular imaging of solid malignancies. The nuclear- and optical-imaging of CAIX-expressing tumors have received great attention due to their potential for clinical applications. Nuclear imaging is a powerful tool for the non-invasive diagnosis of primary and metastatic CAIX-positive tumors and for the assessment of responses to antineoplastic treatment. Intraoperative optical fluorescence imaging provides improved visualization for surgeons to increase the discrimination of tumor lesions, allowing for safer surgical treatment. Over the past decades, many CAIX-targeted molecular imaging probes, based on monoclonal antibodies, antibody fragments, peptides, and small molecules, have been reported. In this review, we outline the recent development of CAIX-targeted probes for single-photon emission computerized tomography (SPECT), positron emission tomography (PET), and near-infrared fluorescence imaging (NIRF), and we discuss issues yet to be addressed.     

碳酸酐酶在功能化Fe3O4-SiO2核壳纳米颗粒上的固定及其性能表征研究

以Fe3O4纳米颗粒为核,正硅酸乙酯（TEOS）为前驱体,通过St?ber法合成了Fe3O4-SiO2核壳纳米颗粒。以γ-缩水甘油醚氧丙基三甲氧基硅烷（GPTMS）为改性剂对纳米颗粒进行改性,得到了环氧基功能化的Fe3O4-SiO2核壳纳米颗粒,随后将碳酸酐酶（carbonic anhydrase,CA）共价固定到纳米颗粒上,以实现CA固定化。采用傅立叶红外（FTIR）、透射电镜（TEM）等手段对载体材料进行表征;以对硝基苯酚乙酸酯（p-NPA）为底物,采用紫外分光光度仪测试酶活力,并对固定化酶进行了性能表征。试验结果表明,固定化酶的热稳定性、贮藏稳定性和循环使用性均优于同等条件下的游离酶,其在循环使用10次后仍保持84.2%的相对酶活力。