盐酸阿霉素聚乳酸纳米粒的制备及大鼠体内药动学研究

目的 优化盐酸阿霉素聚乳酸纳米粒（DOX-PLA-NPs）的制备工艺,并对其理化性质、体外释放及大鼠体内药动学进行研究。方法 采用改良的复乳-溶剂挥发法制备DOX-PLA-NPs,正交设计优化其处方工艺,对其纳米粒形态、粒径、Zeta电位、包封率与载药量进行测定。以DOX原药为对照组,考察DOX-PLA-NPs的体外释药特性及大鼠尾静脉给药后的体内药动学参数。结果 DOX-PLA-NPs外观圆整,平均粒径为（125.67±3.80） nm、Zeta电位为（-35.97±1.58） mV、包封率和载药量分别为（81.23±1.46）%,（10.29±0.63）%。体外释放结果显示,DOX经纳米粒包裹后,具明显的缓释作用。DOX原药和纳米粒的体内药动学过程均符合开放式二室模型,t_1/2β分别为（1.15±0.175） h、（6.43±2.12） h,CL分别为（174.76±47.22） h·L^-1、（30.68±11.86） h·L^-1,AUC_0→t分别为（6.01±1.61）μg·h·L^-1、（36.04±13.72）μg·h·L^-1。结论 制备的盐酸阿霉素聚乳酸纳米粒粒径较小、包封率较高,具明显的缓释作用,并能提高药物的生物利用度。     

星点设计-效应面法优化羟基喜树碱/mPEG-S-S-C18纳米粒的制备工艺

目的 制备载羟基喜树碱（hydroxycamptothecin,HCPT）还原响应mPEG-S-S-C18纳米粒,采用星点设计-效应面法筛选优化制备工艺。方法 采用乳化-溶剂挥发法制备HCPT/mPEG-S-S-C18纳米粒,应用单因素法考察投药量、水相/油相体积比、超声功率以及超声时间对载药纳米粒包封率和载药量的影响。在此基础上,以包封率和载药量作为评价指标,采用Design-Expert V8.0.6软件进行星点设计,优化载药纳米粒的制备工艺。结果 优化获得的HCPT/mPEG-S-S-C18纳米粒制备工艺投药量为1.0 mg,水相/油相体积比为4.56∶1,超声功率为562.5 W。该工艺制备的载药纳米粒包封率为（58.14±1.04）%,载药量为（3.46±0.22）%,平均粒径为（322.9±9.52） nm,多分散性指数为0.195±0.05,Zeta电位为（-17.5±2.11） mV。结论 乳化-溶剂挥发法适用于制备HCPT/mPEG-S-S-C18纳米粒,星点设计-效应面法可优化获得载药纳米粒的最佳制备工艺,所得的载药纳米粒包封率和载药量较高,所建立的数学模型预测性良好。     

季铵化壳聚糖胰岛素纳米粒的制备、处方优化及其初步药效学实验

目的 采用正交设计试验优化载胰岛素季铵化壳聚糖纳米粒的处方工艺,并初步考察其降糖效果。 方法 用离子交联法制备载胰岛素的季铵化壳聚糖纳米粒,用正交试验确定其最佳处方工艺。用透射电子显微镜观察纳米粒的表面形态;用粒径/Zeta电位仪测定纳米粒的粒径和Zeta电位;用高效液相色谱(HPLC)法测定纳米粒的包封率、载药量及体外释放情况。对糖尿病大鼠皮下注射给药,对其药效学进行初步考察。 结果 制得的纳米粒呈球形,分布均匀;平均粒径(63.26±1.88) nm;Zeta电位(33.1±0.3) mV;包封率(37.92±2.11)%;载药量(5.42±0.3)%;24 h累计释放率63.83%。皮下注射给药8 h,糖尿病大鼠血糖较单纯注射胰岛素组下降平缓,且药效持久。 结论 优化后的载胰岛素的季铵化壳聚糖纳米粒形态较好、粒径较小,为研究胰岛素的新型给药途径奠定了基础。     

载阿霉素透明质酸聚合物纳米粒的构建及其体外性质研究

目的 合成透明质酸（HA）接枝单油酸甘油酯（GMO）两亲性聚合物HGO,并研究其所制备载阿霉素（DOX）纳米粒的理化性质及体外抗肿瘤效果。方法 HA与GMO通过酯化反应制得载体聚合物HGO,通过核磁共振波谱法及红外光谱法对其进行结构表征;采用芘荧光探针法测定聚合物临界聚集浓度（CAC）。采用透析法制备聚合物HGO载阿霉素（DOX@HGO）纳米粒,并对其进行粒径分布、Zeta电位及微观形态的表征;通过检测其在不同离子强度、不同pH条件下的粒径变化考察纳米粒的体外稳定性;考察DOX@HGO纳米粒在不同pH条件下的体外释放行为;CCK-8法考察DOX@HGO纳米粒对MDA-MB-231细胞的体外抑瘤效果;并通过荧光显微镜研究MDA-MB-231细胞对DOX溶液、DOX@HGO纳米粒的摄取能力,以及HA预处理对DOX@HGO纳米粒摄取的影响。结果 成功制得两亲性聚合物HGO,聚合物HGO中GMO的取代度为15.8%,CAC为0.023 mg·mL^-1。DOX@HGO纳米粒呈规则的球形,平均粒径为（130.800±1.709）nm,平均电位为（-32.600±0.153）mV,包封率和载药量分别为（98.65±0.74）%和（33.03±0.17）%,在不同离子强度下、模拟胃肠液中表现出良好的稳定性;DOX@HGO纳米粒的体外释放表现出pH依赖性。体外抗肿瘤活性实验表明,DOX@HGO纳米粒对MDA-MB-231细胞的生长具有较好的抑制作用;与DOX溶液比较,DOX@HGO纳米粒显著增加肿瘤细胞对于DOX的摄取（P＜0.05） ,HA预处理显著减少肿瘤细胞对DOX@HGO的摄取（P＜0.05）。结论 所构建的DOX@HGO纳米粒具有良好的理化性质,并且具有一定的pH敏感性及靶向抗肿瘤细胞的能力,是具有应用潜力的药物载体。     

米托蒽醌固体脂质纳米粒的制备与处方优化

目的：制备米托蒽醌固体脂质纳米粒并优化其处方组成。方法：采用薄膜蒸发-超声分散法制备米托蒽醌固体脂质纳米粒。以包封产率、载药量和体外释药为考察指标，中心组合设计优化处方中米托蒽醌、磷脂、山嵛酸甘油酯和硬脂酸聚烃氧酯（S-40）的组成。结果：米托蒽醌固体脂质纳米粒的最优处方组成为米托蒽醌0．2％，磷脂3．0％，山嵛酸甘油酯1．0%，S-400．5％，注射用水加至10mL；优化处方的各指标值依次为包封产率（87．2±2．2）％，载药量（4．2±0．1）％，Q2h为（7．6±0．2）%，Q24h为（25．9±0．8）％，t50为（5．3±1．1）d和t90为（28．4±4．5）d。结论：薄膜蒸发-超声分散法适于制备米托蒽醌固体脂质纳米粒，优化后的各指标值均接近预测值。     

活性氧/谷胱甘肽双重响应型紫杉醇前药纳米粒的制备与评价

目的 设计具有活性氧/谷胱甘肽双重响应的紫杉醇前药纳米粒(ProPTX-SS-NPs),为紫杉醇的应用提供新思路和新方法。方法 以粒径和PDI为指标,考察前药纳米粒的最佳制备方法和工艺;通过电镜观察前药纳米粒的形貌并对其粒径、电位、包封率、载药量等进行考察;考察纳米粒在活性氧和谷胱甘肽环境下的体外释放特性;通过细胞试验考察前药纳米粒的体外细胞毒性和细胞摄取情况。结果 采用最佳工艺制备的纳米粒粒径为(130.20±2.18) nm,分散系数为0.12±0.01,Zeta电位为(–8.45±0.01) mV,载药量为(10.27±1.36)%,包封率为(93.22±2.20)%。前药纳米粒具有良好的活性氧和谷胱甘肽响应特性,并且能够显著抑制MCF-7、HepG2和MDA-MB-231增殖。其对MDA-MB-231细胞的抑制作用最为显著,半数抑制浓度IC₅₀(0.71±0.11)μmol·L^-1,而PTX的IC₅₀为(22.38±3.27)μmol·L^-1。结论ProPTX-SS-NPs具有良好的肿瘤微环境响应性能,具备显著的抗肿瘤活性,是一种极具潜力和应用前景的抗肿瘤纳米系统。     

盐酸环丙沙星壳聚糖纳米粒原位凝胶的制备与评价

目的 制备盐酸环丙沙星壳聚糖纳米粒原位凝胶，并评价其抑菌及创面愈合效果。方法 采用复乳法制备盐酸环丙沙星壳聚糖纳米粒，采用2因素2水平全因子析因实验设计考察了壳聚糖相对分子质量（X₁）和壳聚糖质量浓度（X₂）对壳聚糖纳米粒的药物包封率（Y₁）、粒径分布（Y₂）、多分散系数（Y₃）和Zeta电位（Y₄）的影响；并以泊洛沙姆407作为凝胶基质制备盐酸环丙沙星壳聚糖纳米粒原位凝胶。通过抑菌圈实验比较盐酸环丙沙星乳膏和盐酸环丙沙星壳聚糖纳米粒原位凝胶对金黄色葡萄球菌和铜绿假单胞菌的抑菌活性；使用无菌活检穿刺针在大鼠背部造成直径为5 mm的皮肤全切除的圆形人工创面，并使用金黄色葡萄球菌和铜绿假单胞菌的培养基感染24 h，建立大鼠创面模型，将模型大鼠随机分为模型组、盐酸环丙沙星乳膏组和盐酸环丙沙星壳聚糖纳米粒原位凝胶组，模型组大鼠创面未接受任何处理，给药组大鼠每2天给药1次，每次给药量均约为1 mg，观察并记录每组大鼠创面脱痂时间和愈合时间。结果 选择低相对分子质量壳聚糖、壳聚糖质量浓度为2.0 mg·mL^-1制备盐酸环丙沙星壳聚糖纳米粒，其中盐酸环丙沙星质量浓度为50.0 mg·mL^-1，其包封率为（85.3±0.9）%，平均粒径为（354.7±15.7）nm，PDI为0.357±0.014，Zeta电位为（22.2±0.5）mV，呈球状分布；盐酸环丙沙星壳聚糖纳米粒原位凝胶和盐酸环丙沙星乳膏对金黄色葡萄球菌的抑菌圈直径分别为（38.4±0.2）、（29.2±0.3）mm，对铜绿假单胞菌抗菌圈直径分别为（41.3±0.6）、（32.1±0.1）mm；大鼠创面给予盐酸环丙沙星壳聚糖纳米粒原位凝胶后，其脱痂时间和愈合时间均较模型组和盐酸环丙沙星乳膏组显著缩短（P<0.05）。结论 成功制备盐酸环丙沙星壳聚糖纳米粒原位凝胶，其可以抑制创面细菌繁殖、加速伤口愈合。     

葫芦素B-聚乳酸-羟基乙酸共聚物载药纳米粒的构建与药效学评价

目的 以聚乳酸-羟基乙酸共聚物（PLGA）作为纳米制剂载体材料将葫芦素B制备成纳米粒,并考察其对HepG2肝癌细胞的抑制效果。方法 使用乳化溶剂蒸发法制备葫芦素B-PLGA载药纳米粒,以PLGA浓度（X₁）、PVA浓度（X₂）和药物浓度（X₃）作为考察因素,以载药纳米粒的粒径大小（Y₁）和包封率（Y₂）作为评价指标,应用中心复合设计-效应面法优化葫芦素B-PLGA载药纳米粒处方;测定了纳米粒的粒径分布和Zeta电位值,通过透射电镜观察其微观形态,并考察了葫芦素B-PLGA载药纳米粒的体外药物释放特性;比较了葫芦素B与葫芦素B-PLGA载药纳米粒对HepG2肝癌细胞的抑制效果。结果 葫芦素B-PLGA载药纳米粒的最优处方组成为：PLGA浓度为9.0%,PVA浓度为2.0%,药物浓度为4.5%,制备的纳米粒粒径为（145.4±15.8） nm,Zeta电位值为（-7.6±0.8） mV;透射电镜下可观察到纳米粒表面光滑,分布均匀;葫芦素B-PLGA载药纳米粒释药前期出现突释,后期平缓,48 h药物释放达到86%;葫芦素B-PLGA载药纳米粒对HepG2肝癌细胞的抑制作用显著高于葫芦素B。结论 葫芦素B-PLGA载药纳米粒可延缓药物释放,提高对HepG2肝癌细胞的抑制活性,为进一步临床研究奠定实验基础。     

尼莫地平PEG-PLGA纳米粒的制备及其处方优化

目的：负载尼莫地平的聚乙二醇修饰的聚乳酸-羟基乙酸共聚物[poly （ethylene glycol-poly （lactin-co-glycolic acid）,PEG-PLGA）]纳米粒,并对其进行制备工艺、质量评价以及体外释放等相关性研究。方法：以PEG-PLGA为药物载体,采用乳化溶剂挥发法成功制备尼莫地平载药纳米粒。单因素实验和响应面法设计优化处方工艺,透射电子显微镜观察纳米粒形态,激光粒度仪测定其粒径和Zeta电位,HPLC法测定其包封率及载药量并考察其体外释药特性。结果：制备的尼莫地平纳米粒外观呈实心球体,大小均匀且分散性良好;平均粒径为（183.2±3.30） nm,PDI为（0.115±0.049）,Zata电位为（-11.78±2.16） mV;平均包封率为84.99%,平均载药量为2.45%;尼莫地平原料药在4 h时基本释放完全（达到95%左右）,而尼莫地平纳米粒在4 h时释放仅为43.9%,在第24 h时累计释放度达到（83.66±2.57）%。与对照组相比,制剂组释放缓慢,符合实验设计缓释的要求。结论：本实验成功制备了尼莫地平PEG-PLGA纳米粒,其体外释药具有明显缓释特征,为心脑血管疾病的治疗奠定了基础。     

醋酸泼尼松龙固体脂质纳米粒的试制

采用乳化蒸发-低温固化法制备了醋酸泼尼松龙固体脂质纳米粒,采用正交实验设计优化处方,并考察了理化性质。结果表明,所得制品呈类球状,粒径较均匀,平均粒径为（187±15）nm,载药量为4.8%,包封率79.4%。     

Efficacy of gut lavage,hemodialysis, and hemoperfusion in the therapy of paraquat or diquat intoxication

Clinical and in vitro investigations were carried out to test the efficacy of gut lavage, hemodialysis, and hemoperfusion in the treatment of poisoning with paraquat or diquat. In a patient suffering from diquat intoxication 130 times more diquat was removed by gut lavage 30 h after ingestion than was removed by complete aspiration of the gastric contents.Determination of in vitro clearances for paraquat and diquat by hemodialysis showed that, at serum concentrations of 1–2 ppm, such as are frequently encountered in poisoning in man, toxicologically relevant quantities of herbicide cannot be removed from the body. At a concentration of 20 ppm, on the other hand, hemodialysis proved to be effective, the clearance being 70 ml/min at a blood flow rate of 100 ml/min. The efficacy of hemoperfusion with coated activated charcoal was on the whole better. Especially at concentrations around 1–2 ppm, the clearance values for hemoperfusion were some 5–7 times higher than those for hemodialysis.In a patient suffering from paraquat poisoning, both hemodialysis as well as hemoperfusion were carried out. The in vitro results could be confirmed: At serum concentrations of paraquat less than 1 ppm no clearance could be obtained by hemodialysis while by hemoperfusion with activated charcoal quite high clearance values were measured and the serum level dropped down to zero.

---

Zusammenfassung Klinische Untersuchungen und Laboratoriumsversuche wurden durchgeführt, um die Wirksamkeit von Darmspülung, Hämodialyse und Hämoperfusion bei Paraquat- und Deiquat-Vergiftungen zu prüfen.Bei einem Patienten wurde 30 Std nach Deiquat-Aufnahme durch Darmspülung 130mal mehr Deiquat entfernt als durch vollständige Aspiration des Mageninhaltes. In vitro-Versuche ergaben, daß bei Blutserumkonzentrationen von 1–2 ppm, die bei Vergiftungen oft gemessen werden, durch Hämodialyse keine toxikologisch relevanten Paraquat- oder Deiquat-Mengen entfernt werden können. Dagegen erwies sich die Hämodialyse bei 20 ppm und einer Blutumlaufgeschwindigkeit von 100 ml/min mit einer Clearance von 70 ml/min als wirksam. Die Hämoperfusion mit beschicheter Aktivkohle war in diesen Versuchen aber eindeutig überlegen, denn insbesondere bei Konzentrationen um 1–2 ppm waren die Clearance-Werte 5–7mal höher als bei der Hämodialyse.Die in vitro-Ergebnisse wurden bei einem Patienten mit einer Paraquat-Vergiftung bestätigt: Bei Konzentrationen unter 1 ppm war die Hämodialyse wirkungslos, während durch Hämoperfusion relativ hohe Clearance-Werte erreicht wurden, so daß der Serumspiegel rasch unter die Nachweisgrenze abfiel.

     

In vitro studies on sterically stabilized liposomes (SL) as enzyme carriers in organophosphorus (OP) antagonism

This study describes a new approach for organophosphorous (OP) antidotal treatment by encapsulating an OP hydrolyzing enzyme, OPA anhydrolase (OPAA), within sterically stabilized liposomes. The recombinant OPAA enzyme was derived from Alteromonas strain JD6. It has broad substrate specificity to a wide range of OP compounds: DFP and the nerve agents, soman and sarin. Liposomes encapsulating OPAA (SL)* were made by mechanical dispersion method. Hydrolysis of DFP by (SL)* was measured by following an increase of fluoride ion concentration using a fluoride ion selective electrode. OPAA entrapped in the carrier liposomes rapidly hydrolyze DFP, with the rate of DFP hydrolysis directly proportional to the amount of (SL)* added to the solution. Liposomal carriers containing no enzyme did not hydrolyze DFP. The reaction was linear and the rate of hydrolysis was first order in the substrate. This enzyme carrier system serves as a biodegradable protective environment for the recombinant OP-metabolizing enzyme, OPAA, resulting in prolongation of enzymatic concentration in the body. These studies suggest that the protection of OP intoxication can be strikingly enhanced by adding OPAA encapsulated within (SL)* to pralidoxime and atropine.     

Pharmacological treatment of COVID-19: an opinion paper

The precocity and efficacy of the vaccines developed so far against COVID-19 has been the most significant and saving advance against the pandemic. The development of vaccines has not prevented, during the whole period of the pandemic, the constant search for therapeutic medicines, both among existing drugs with different indications and in the development of new drugs. The Scientific Committee of the COVID-19 of the Illustrious College of Physicians of Madrid wanted to offer an early, simplified and critical approach to these new drugs, to new developments in immunotherapy and to what has been learned from the immune response modulators already known and which have proven effective against the virus, in order to help understand the current situation.     

Aquatic Insects and Trace Metals: Bioavailability,Bioaccumulation, and Toxicity

Abstract

The uptake of metals from food and water sources by insects is thought to be additive. For a given metal, the proportions taken up from water and food will depend both on the bioavailable concentration of the metal associated with each source and the mechanism and rate by which the metal enters the insect. Attempts to correlate insect trace metal concentrations with the trophic level of insects should be made with a knowledge of the feeding relationships of the individual taxa concerned. Pathways for the uptake of essential metals, such as copper and zinc, exist at the cellular level, and other nonessential metals, such as cadmium, also appear to enter via these routes. Within cells, trace metals can be bound to proteins or stored in granules. The internal distribution of metals among body tissues is very heterogeneous, and distribution patterns tend to be both metal and taxon specific. Trace metals associated with insects can be both bound on the surface of their chitinous exoskeleton and incorporated into body tissues. The quantities of trace meals accumulated by an individual reflect the net balance between the rate of metal influx from both dissolved and particulate sources and the rate of metal efflux from the organism. The toxicity of metals has been demonstrated at all levels of biological organization: cell, tissue, individual, population, and community. Much of the literature pertaining to the toxic effects of metals on aquatic insects is based on laboratory observations and, as such, it is difficult to extrapolate the data to insects in nature. The few experimental studies in nature suggest that trace metal contaminants can affect both the distribution and the abundance of aquatic insects. Insects have a largely unexploited potential as biomonitors of metal contamination in nature. A better understanding of the physico-chemical and biological mechanisms mediating trace metal bioavailability and exchange will facilitate the development of general predictive models relating trace metal concentrations in insects to those in their environment. Such models will facilitate the use of insects as contaminant biomonitors.     

Alzheimer’s disease – current trends in basic and clinical research. Highlights from the 16th ECNP

Advances in the molecular biological knowledge of neuronal nicotinic acetylcholine receptors (nAChRs) have led to a growing interest by the pharmaceutical industry in the development of novel compounds that selectively modulate nAChR function. The ability of (-)-nicotine, an activator of nAChRs, to enhance attentional aspects of cognition in animals and humans, to exert neuroprotective and anxiolytic-like effects, and presumably to mediate the negative correlation between smoking and Alzheimer's (and Parkinson's) Disease, has focused interest on the potential therapeutic utility of modulators of nAChR function for treatment of some of the deficits associated with these progressive, neurodegenerative conditions. Numerous compounds are known which activate nAChRs and which might serve as lead compounds toward the development of such agents. The pharmacologic diversity of neuronal nAChR subtypes suggests the possibility of developing selective compounds which would have more favourable side-effect profiles than existing agents. This broader class of agents, collectively called cholinergic channel modulators (ChCMs), is anticipated to encompass compounds which would have more favourable side-effect profiles than existing agents, which generally exhibit low selectivity. This selectivity may be achieved by preferentially activating some subtypes of nAChRs (i.e., Cholinergic Channel Activators, ChCAs) or inhibiting the function of other subtypes (Cholinergic Channel Inhibitors, ChCIs). An overview of the biology of nAChRs and the rationale for the use of ChCMs for the treatment of dementia related to neurodegenerative diseases are presented, followed by a discussion of lead compounds and compounds under consideration for clinical evaluation.     

Steroidal sapogenin contents in some domestic plants

In order to find out the values of the steroid resources for the future use. the compositions and contents of steroidal sapogenins from 13 domestic plants have been investigated. As a result,Dioscorea nipponica, D. quinqueloba andSmilax china were found to have large amount of diosgenin. And pennogenin inTrillium kamtschaticum andParis verticillata, yuccagenin inAllium fistulosum, hecogenin inAgave americana and neochlorogenin inSolanum nigum were appeared to be major steroidal sapogenins.