银模板法制备中空金纳米粒及其质量评价

目的 制备并表征一种可用于载药的中空金纳米粒载体.方法 以银纳米粒为模板,通过模板置换法使氯金酸与银纳米粒反应生成金壳,并裹覆于银纳米粒表面,最后形成一种中空结构的金纳米粒,通过单因素考察对中空金纳米粒的制备进行初步探究.使用透射电镜对中空金纳米粒进行结构观察,紫外全波长扫描考察其吸收情况,体外光热转化试验考察载体的光热转化能力,以及四甲基偶氮唑蓝(MTT)试验考察载体细胞毒性.结果 氯金酸的加入方式为逐滴加入,制备温度为60 ℃,氯金酸(25 mmol·L-1)的加入量为128 μL,中空金纳米的粒径为35~55 nm,壳厚4~6 nm,最大吸收波长750~800 nm,具备较强的光热转化能力.细胞毒性结果表明,中空金纳米粒对人皮肤成纤维细胞(HDF)没有明显的细胞毒性.结论 经过优化制备得到的中空金纳米粒具备较好的中空结构,以及优异的光热转化能力和较低的细胞毒性.     

载阿霉素金纳米粒的制备和细胞毒性研究

目的 构建载阿霉素（DOX）的甲氧基聚乙二醇（mPEG）修饰的金纳米粒AuNPs-mPEG@DOX，以降低DOX的毒副作用。方法 制备AuNPs-mPEG@DOX，通过粒径、电位和紫外可见光吸收光谱（UV-Vis）进行表征。考察连接巯基的DOX（HS-DOX）投药浓度对AuNPs-mPEG@DOX吸附率和载药量的影响。建立未吸附HS-DOX含量测定的高效液相色谱法（HPLC），对专属性、线性、精密度、稳定性和加样回收率进行考察。采用CCK-8法检测AuNPs-mPEG@DOX对MCF-10A和MCF-7细胞的毒性作用。结果 成功制备了AuNPs-mPEG@DOX，粒径为（46.12±0.49） nm，电位为（18.60±1.51） mV，最大吸收波长为530 nm。建立了可用于检测AuNPs-mPEG@DOX未吸附HS-DOX含量的HPLC方法，测定最佳投药浓度11.18 μg/ml，HS-DOX条件下的吸附率为（9.21±2.88）%，载药量为（2.01±0.62）%。细胞毒性实验表明AuNPs-mPEG@DOX可明显降低DOX对正常乳腺细胞的毒副作用；DOX在≥4.75 μmol/L时，AuNPs-mPEG@DOX与游离DOX对乳腺肿瘤细胞的细胞毒性作用一致。结论 AuNPs-mPEG@DOX可有效降低DOX的毒副作用，为后续AuNPs连接药物降低其毒副作用的研究提供参考。     

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。     

负载阿霉素的黄芩苷纳米粒的制备及其体外性能评价

目的 制备负载阿霉素的黄芩苷纳米粒（DOX/SA-SS-BAI NPs）,并评价其体外性能。方法 构建以胱胺为连接臂的海藻酸钠–黄芩苷聚合物,并负载阿霉素,得到DOX/SA-SS-BAI NPs。对DOX/SA-SS-BAI NPs的理化性质进行表征;采用HepG2细胞进行MTT实验验证其细胞毒性。结果 DOX/SA-SS-BAI NPs粒径为（158.2±2.8）nm,PDI为（0.241±0.008）,Zeta电位为（−24.1±0.3）mV,包封率为（64.34±0.25）%,载药量为（16.22±0.06）%。体外释放显示载药纳米粒具有良好的还原响应性;MTT实验证明DOX/SA-SS-BAI NPs对HepG2细胞具有良好的抑制作用;细胞摄取实验表明DOX/SA-SS-BAI NPs在HepG2细胞内较快地释放阿霉素。结论 制备的DOX/SA-SS-BAI NPs具有较好的理化性质和体外抗癌作用。     

硫化铜纳米粒的处方工艺优化与体外评价

目的 为避免硫化铜（CuS）纳米粒体内蓄积,制备并优化CuS纳米粒,分析粒径影响因素并评价其光热性能。方法 在单因素考察基础上,采用星点设计-响应面法进一步分析各因素对CuS纳米粒粒径的影响,获得最佳处方工艺。考察优选的纳米粒的理化性质,包括形态、粒径稳定性、光热转换性能及光热稳定性。采用CCK-8法评价CuS纳米粒对4T1乳腺癌细胞以及HK2肾细胞的毒性,并考察其光热效应对4T1乳腺癌细胞的杀伤效果。结果 CuS纳米粒优选处方工艺的水合粒径为（10.53±1.63）nm;透射电镜下显示其粒径为（3.10±0.81）nm;所优选的CuS纳米粒具有良好的粒径稳定性,良好的光热转换性能与光热稳定性。细胞毒研究显示,所优选的CuS纳米粒分别在100 μg/ml及150 μg/ml浓度范围内,对4T1乳腺癌细胞以及HK2肾细胞均无显著的细胞毒性,且其对4T1乳腺癌细胞具有显著的光热杀伤效果。结论 所制备的CuS纳米粒实际粒径小于6 nm且具有良好的光热效应,有望解决CuS纳米粒体内蓄积的问题,使其更好地应用于抗肿瘤治疗。     

巯基化阿霉素的两种合成方法的比较

目的：探索合成供金纳米粒载药系统研究用模型药物巯基化阿霉素的可行方法。方法分别采用2-亚氨基硫烷盐酸盐（2-IT）法和琥珀酰亚胺-S-乙酰基硫代乙酸酯（SATA）法合成巯基阿霉素,通过高效液相色谱（HPLC）、飞行时间质谱（MS-ESI）及核磁共振氢谱（1 H NMR）验证巯基阿霉素的合成,并考察反应物摩尔比、反应时间等因素对合成巯基阿霉素的影响。结果1 H NMR确证DOX-SATA出现了与硫酯基团相连的质子信号,表明新合成的化合物中含有硫酯基团。 HPLC及MS-ESI结果显示,两种方法均能合成巯基阿霉素,2-IT法生成的巯基阿霉素,随着反应时间延长易发生环化,形成环化巯基阿霉素。 SATA试剂法合成巯基阿霉素过程中不易发生副反应,合成的巯基阿霉素较为稳定。结论通过两种方法的比较,SA-TA法合成巯基阿霉素的方法较为可行。     

共载阿霉素和依克立达的PLGA纳米粒的制备及表征

目的 建立药物测定方法,并制备共载阿霉素和依克立达的PLGA纳米粒。方法 利用紫外分光光度法（UV）和高效液相色谱法（HPLC）分别建立阿霉素和依克立达的测定方法;采用纳米沉淀法制备共载纳米粒,通过调节两药的投药比,优化处方,考察纳米粒的粒径、形态、包封率、载药量以及体外释放。结果 阿霉素在1~40 μg/ml浓度范围内线性关系良好,标准曲线回归方程为A=0.021C+0.002,r=0.999 5; 依克立达在0.5~100 μg/ml浓度范围内线性关系良好,标准曲线回归方程为A=120 742.462 6C+1 974.570 4,r=1.000 0;通过处方优化,共载纳米粒的粒径约为50 nm,分布均一,呈圆形,阿霉素和依克立达的包封率分别为56.58%、51.66%,载药量分别为1.48%、1.85%,两药摩尔比约为1：1;体外释放缓慢。结论 分别建立了方便快捷、结果准确、重复性好的阿霉素和依克立达的检测方法,并且制备了分散性好、粒径较小的纳米粒,为后续实验提供基础。     

用疏水改性的白及多糖制备载紫杉醇纳米粒并对其表征

目的 讨论白及多糖作为药物递送载体的可行性。方法 制备疏水性胆甾醇琥珀酰基白及多糖（CHSB）后,以紫杉醇（PTX）为模型药物,采用透析法制备载药纳米粒子,然后在透射电镜（TEM）下观察其形态;用动态光散射仪（DLS）检测其粒径、粒径分布和Zeta电位;用高效液相色谱法（HPLC）测定其包封率和载药量,并考察其体外释放情况;采用差示量热扫描法（DSC）确证药物在载药纳米粒子中的存在形式;采用MTT法考察纳米粒子的体外抗肿瘤活性,用荧光标记法观察肝癌细胞QGY-7703对纳米粒子的摄取情况。结果 制备的纳米粒呈规则球形,粒度分布均匀,药物包载于纳米粒内部,载药量和包封率在一定范围受CHSB的影响,载药纳米粒对肝癌细胞的杀伤性强于游离药物,在细胞内可观察到罗丹明B标记的纳米粒呈现的荧光。结论 CHSB作为难溶性药物载体具有较高的可行性,因此可作为一种极具潜力的纳米载体材料。     

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。     

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。     

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.     

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.     

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.