抗人肝癌免疫毫微粒的制备及体外免疫学性质的鉴定

目的 构建人肝癌特异的免疫毫微粒,探讨免疫微粒体外对靶细胞的特异性结合特性及杀伤性活。方法 采用异型双功能交联剂ＳＰＤＰ将人肝癌单抗ＨＡｂ１８与载米托蒽醌的白蛋白毫微粒化学偶联,构建人肝特异的免疫毫微粒;采用玻片凝集试验,免疫荧光法及荧光染色阻断法,花环形成实验及花环形成阻断实验,扫描电镜,＾３Ｈ－ＴｄＲ掺入试验证明抗体与载药毫微粒偶联及免疫毫微粒能特异性结合并杀伤靶细胞ＳＭＭＣ－７７２１人肝癌株     

示踪剂盐酸米托蒽醌在甲状腺癌根治术中的应用

目的 研究示踪剂盐酸米托蒽醌在甲状腺癌根治术中对淋巴结染色率、淋巴结示踪率及甲状旁腺误切率的影响。方法 选取2022年2月至8月于我院行甲状腺癌根治术的106例患者,随机分为对照组和观察组,每组53例。记录患者淋巴结染色率、示踪持续成功率。比较2组患者淋巴结清除数目、甲状旁腺误切率,及2组患者术前及术后第1天、第3天血钙和甲状旁腺激素（PTH）。结果 观察组患者淋巴结染色率为90.1%,示踪持续成功率为100%。2组患者淋巴结清除数目,甲状旁腺误切率,术后第1天、第3天血钙和PTH比较差异均有统计学意义（P<0.05）。结论 示踪剂盐酸米托蒽醌具有良好的安全性和淋巴结示踪性,能降低甲状旁腺误切率,值得临床推广应用。     

盐酸表阿霉素纳米靶向制剂的缓释效应

背景：盐酸表阿霉素是一种广谱抗生素,目前临床使用的不足多为药物释放快、目标组织药物浓度低,静脉给药后广泛分布于体内各种组织器官,不良反应明显。 目的：针对盐酸表阿霉素临床应用的不足,制备盐酸表阿霉素纳米靶向注射制剂。 方法：以叶酸偶联牛血清白蛋白为载体,采用乳化-高压匀质法,制备盐酸表阿霉素纳米靶向注射制剂,以激光粒度分析仪测定纳米颗粒的粒径大小、粒径分布及Zeta电位,扫描电镜观察纳米颗粒的表面形态,高效液相色谱法分析白蛋白负载盐酸表阿霉素纳米制剂的包封率、载药量和释药性能。 结果与结论：制备的盐酸表阿霉素纳米粒外观呈均匀球型,粒径分布较窄,平均粒径为(157.73±     0.40) nm,平均 Zeta 电位为(-30.85±0.43) mV,载药量 22.78%,包封率可达96.24%。体外模拟释药结果表明药物释放曲线分为两个阶段,突释阶段微球释药量在24 h内达42.6%,缓释阶段纳米粒释药持续时间长,在112 h 时释药量达 84.1%,载药纳米粒的药物释放速率持续稳定。结果表明乳化结合高压匀质法制备的盐酸表阿霉素纳米靶向制剂粒径均匀,粒径范围分布窄,载药量和包封率高,具有一定的缓释作用。     

乳酸-羟基乙酸共聚物载药纳米粒的制备及体外释药性

背景：医用纳米粒作为药物传递的新型载体,目前已经成为医药领域研究的重点。 目的：构建以生物可降解材料乳酸-羟基乙酸共聚物为载体,负载抗肿瘤药物5-氟尿嘧啶的载药纳米粒。 方法：利用复乳-溶剂挥发法制备乳酸-羟基乙酸共聚物载药纳米粒。场发射扫描电子显微镜观察纳米粒表面形态;激光粒度分析仪测定粒径分布并计算成球率;紫外分光光度计测定5-氟尿嘧啶载药量、包封率,并对体外释药进行评估。 结果与结论：纳米粒呈球性,平均粒径为(186±14) nm,成球率、载药量和包封率分别为70.8%、6.6%、28.1%,体外释药有突释现象,24 h内5-氟尿嘧啶累积释药量达36.2%,10 d达83.6%。提示成功制备乳酸-羟基乙酸共聚物载药纳米粒,其具有缓释效应。     

盐酸米多君原料药含量测定方法的改进

目的:建立革除汞盐的氢氧化钠电位滴定法测定盐酸米多君原料药的含量。方法:采用Mettler Toledo DG111-SC(pH 0-14,0-70℃)复合电极,以乙醇为溶剂,采用氢氧化钠滴定液(0.1mol·L~(-1))滴定,并与国家药品试行标准YBH09272004中含量测定原方法进行了比较。结果:空白无干扰,重复性良好,成功革除醋酸汞,两种测定方法含量结果比较无明显差异。结论:建立的新方法革除了醋酸汞,滴定突跃明显,方法简单易行,重复性好。     

N-(2-羟丙基)甲基丙烯酰胺聚合物-米托蒽醌接合物荷瘤小鼠体内分布的研究

本实验成功合成N (2 羟丙基 )甲基丙烯酰胺聚合物 (N (2 hydroxypropyl)methacrylamide ,HPMA) 米托蒽醌接合物。以小鼠皮下接种艾氏癌实体型作为肿瘤模型 ,考察游离药物与接合物在动物体内分布及药代动力学行为。研究结果表明 ,接合物较之游离药物在荷瘤小鼠体内的分布明显不同 ,表现为具有较长的血浆半衰期及较强的肿瘤趋向性 ,证实了HPMA聚合物对实体瘤具有“增强的透过及滞留效应”。同时 ,接合物在心脏中的分布明显减少 ,降低了原药的毒副作用。从而更进一步说明 ,将具有仲氨基的米托蒽醌连接于HPMA聚合物 ,较原药而言 ,可提高肿瘤组织的分布 ,用于临床实体瘤治疗 ,具有一定的潜力。     

载药敷料体外细胞毒性及释药性评价

目的评价载药敷料体外细胞毒性和释药性。方法按国标GB/T14233.2-2005,规定的MTT法操作,通过光学倒置显微镜观察L929细胞形态和增殖状况定性分析载药敷料体外细胞毒性,通过吸光度(OD)值计算细胞相对增殖率定量分析载药敷料体外细胞毒性等级;以磷酸盐缓冲液(PBS7.4)为溶出介质,(32±5)℃模拟人体表皮温度,于1/6、1/2、1、3、16、24、36和48h不同时间点取样,采用紫外分光光度法测定载药敷料体外累积释放性能。结果该载药敷料浸提原液中的细胞形态正常,贴壁生长良好,细胞平均相对增殖率(RGR)为91.25%,毒性为1级。载药敷料的释药动力学符合Higuchi方程:拟合方程为Mt/M∞=0.3271t0.239。结论该载药敷料细胞相容性良好,并具有一定的缓释性能。     

脑主动靶向载内吗啡肽纳米粒的制备及其镇痛作用的实验研究

目的 制备新型载镇痛药物内吗啡肽的耦联脑主动靶向抗体OX26纳米粒,研究其镇痛作用.方法 利用新型材料超支化聚甘油-聚乳酸-聚乙醇酸(HBPG-PLGA)通过复合乳液法制备载镇痛药内吗啡肽的纳米粒,在其表面耦联转铁蛋白受体单克隆抗体OX26,通过扫描电子显微镜等进行表征;通过尾静脉注射不同配方纳米粒,考察纳米粒透过血-脑屏障及其对慢性坐骨神经结扎(CCI)大鼠模型镇痛作用.结果 载内吗啡肽HBPG-PLGA纳米粒制备方法稳定,在扫描电子显微镜下呈核壳结构,平均粒径(170±20)nm,Zeta电位约-27 mV,与抗体OX26的耦联效果较好,药物可以缓慢释放72 h以上.尾静脉注射耦联抗体OX26载内吗啡肽纳米粒对CCI大鼠具有较好的镇痛作用,未耦联抗体OX26纳米粒只在给药后45 min时间点显示微弱镇痛作用,静脉注射内吗啡肽和空白纳米粒未显示镇痛作用.结论 耦联抗体OX26载内吗啡肽纳米粒制备稳定,达到较好透血-脑屏障作用,对CCI大鼠有良好镇痛作用.     

加载PEDF的多聚体超声微泡研制及其特性

目的:针对血管新生类疾病,研制与色素上皮源因子（PEDF）具有最大结合率的多聚体超声微泡,并明确其理化特性。方法:首先应用声空化方法制备多聚体超声微泡;然后将PEDF按浓度分为0.4、2.0、10.0和50.0 μg /mL组,应用跨膜按梯度法包裹入脂质体,使其与超声微泡结合,进而置于荧光显微镜下观察。应用流式细胞仪检测PEDF与超声微泡的最大结合率,采用荧光分光光度法测定载药超声微泡的包封率及体外释药特性。结果:成功制备了与PEDF最大结合率为（96.14±1.21）%的多聚体超声微泡。脂质体对PEDF的包封率约为（79.20±2.31）%,体外释放浓度于微泡击破后1 min内达到峰值,持续释放10 min后浓度显著减低。结论:与PEDF具有较高结合率的多聚体超声微泡可以作为合适的药物载体,靶向治疗血管新生类疾病。     

载荷角质细胞生长因子聚乳酸-羟基乙酸共聚物纳米缓释微球在组织工程皮肤构建中的应用

背景：聚乳酸-羟基乙酸共聚物具有良好的生物相容性和可降解性性。 目的：制备载荷角质细胞生长因子聚乳酸-羟基乙酸共聚物控释载药系统用于组织工程皮肤。 方法：采用乳化-溶剂挥发法、冷冻干燥法制备载有角质细胞生长因子的聚乳酸-羟基乙酸共聚物纳米微球,并构建组织工程皮肤。扫描电镜、倒置显微镜、纳米粒度分析仪、紫外分光及ELISA法对微球评价其特性。 结果与结论：纳米微球载药量为(14.05±0.56)%,包封率为(59.86±2.38)%,角质细胞生长因子活性保留率(78.26±5.63)%,体外释放30 d的累积释药率达75%以上,微球形态规则圆润。微球形态规整,在脱细胞真皮表面分布均匀,与其联接良好。毛囊干细胞群在荷载聚乳酸-羟基乙酸共聚物纳米微囊脱细胞真皮上生长活跃,细胞形态良好,并呈克隆团生长。说明实验用组织工程材料制备工艺合理,材料相容性好,可用于构建新型组织工程皮肤。     

Antigenicity of the proteins in soy lecithin and soy oil in soybean allergy

BACKGROUND: Soy lecithin and soy oil are usually produced from the hexane extract of soybean. Some of the soybean proteins are included in the extract and are therefore present in small amounts in both soy lecithin and soy oil. The antigenicity of the proteins present in defatted soybean has been studied with respect to soybean allergy, but the antigenicity of those found in the extract is yet to be investigated. OBJECTIVE: The antigenicity of soy lecithin and soy oil proteins with regard to soybean allergy were investigated. METHODS: The proteins present in soy lecithin and soy oil were determined according to already established method and analysed by SDS-PAGE. The IgE- and IgG4-binding abilities of the soy lecithin proteins were investigated by immunoblotting with sera from 30 soybean-sensitive patients, including seven with a positive challenge test. Immunoblotting of soy oil proteins was performed with the sera from some of these patients. RESULTS: In 100 g of sample, the soy lecithin and soy oil contained 2.8 mg and 1.4-4.0 microg of proteins, respectively. The results of SDS-PAGE demonstrated the presence of only three proteins, with molecular weights of about 58-67 kDa in soy oil, and suggested that soy lecithin also contains these proteins. The soy lecithin also contained many proteins besides these. In the soy lecithin, the detection rate of only one protein, with a molecular weight of 31 kDa, by the serum IgE of patients was significantly different compared with controls (detection rate: 40%). The proteins with molecular weights of 58-67 kDa rarely bound to serum IgE. Only one of the patients who presented a positive challenge test had IgE antibodies to soy lecithin proteins. IgG4-binding proteins were found only rarely in soy lecithin. Neither the IgE nor the IgG4 present in the patients' sera reacted to any soy oil protein. CONCLUSION: Proteins present in soy lecithin and soy oil have little antigenicity with regard to soybean allergy.     

Foam-stability test on gastric aspirate and the diagnosis of respiratory-distress syndrome.

Gastric aspirate, collected from 79 infants within 30 minutes of birth, was subjected to the foam-stability test. The lecithin/sphingomyelin ratio was determined in 27. The results were compared with the incidence of respiratory-distress syndrome as determined independently by different investigators. Of the 59 infants with a positive foam-stability test on gastric aspirate, three had transient respiratory distress, and one the respiratory-distress syndrome; 17 of 22 had lecithin/sphingomyelin ratios greater than 2.0. Of nine infants who had intermediate test results, three were normal, four had transient respiratory distress, and two had the respiratory-distress syndrome. In all the 11 infants with negative foam-stability tests the respiratory-distress syndrome developed. The three gastric aspirates tested in this group had lecithin/sphingomyelin ratios of less than 1.5. These data indicate that the foam-stability test on gastric aspirate is a reliable index of fetal lung maturity in infants whose amniotic fluid is not available.     

Identification of IgE-binding proteins in soy lecithin.

BACKGROUND: Soy lecithin is widely used as an emulsifier in processed foods, pharmaceuticals and cosmetics. Soy lecithin is composed principally of phospholipids; however, it has also been shown to contain IgE-binding proteins, albeit at a low level. A few clinical cases involving allergic reactions to soy lecithin have been reported. The purpose of this investigation is to better characterize the IgE-binding proteins typically found in lecithin. METHODS: Soy lecithin proteins were isolated following solvent extraction of lipid components and then separated on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The separated lecithin proteins were immunoblotted with sera from soy-sensitive individuals to determine the pattern of IgE-binding proteins. The identity of IgE-reactive bands was determined from their N-terminal sequence. RESULTS: The level of protein in six lecithin samples obtained from commercial suppliers ranged from 100 to 1,400 ppm. Lecithin samples showed similar protein patterns when examined by SDS-PAGE. Immunoblotting with sera from soy-sensitive individuals showed IgE binding to bands corresponding to 7, 12, 20, 39 and 57 kD. N-terminal analysis of these IgE-binding bands resulted in sequences for 3 components. The 12-kD band was identified as a methionine-rich protein (MRP) and a member of the 2S albumin class of soy proteins. The 20-kD band was found to be soybean Kunitz trypsin inhibitor. The 39-kD band was matched to a soy protein with unknown function. CONCLUSIONS: Soy lecithin contains a number of IgE-binding proteins; thus, it might represent a source of hidden allergens. These allergens are a more significant concern for soy-allergic individuals consuming lecithin products as a health supplement. In addition, the MRP and the 39-kD protein identified in this study represent newly identified IgE-binding proteins.     

Suppression of experimental allergic encephalomyelitis by mitoxantrone

Treatment of rats with a developing or an established lesion of experimental allergic encephalomyelitis (EAE) with mitoxantrone (Novantrone) suppressed the hind limb paralysis associated with the disease. Histopathological examination of the spinal cords of these rats showed that mitoxantrone-treated rats had reduced vascular lesions that are associated with EAE. Spleen cells derived from immunized rats that had been treated in vivo with mitoxantrone did not transfer disease when these cells were administered to naive syngenic recipients. In addition, spleen cells from diseased rats did not transfer EAE lesions when these cells were administered to recipients that had been treated with mitoxantrone. Recipients treated with mitoxantrone were resistant to EAE lesions induced by sensitized cells in a rapid passive transfer system. Finally, when spleen cells from rats with EAE were incubated, in vitro, with mitoxantrone, these cells did not transfer disease to recipients. Thus the present studies indicate that treatment with mitoxantrone can suppress the lesions associated with both the active and passive forms of EAE.     

Interaction of isolated influenza virus hemagglutinin with flat lipid membranes

The interaction of isolated influenza virus hemagglutinin with cell membranes was studied on the model of flat lipid membranes from lecithin and azolectin. Protein molecules were shown to adsorb on the membrane, and permeability of the latter increases when a certain concentration is reached. At the stages of adsorption and penetration into the lipid bilayer the hemagglutinin (HA) showed positive cooperation in interaction with the membranes. At the stage of HA adsorption on the membrane in the presence of M protein, transmembrane interaction occurs between M-protein and HA. It is concluded that molecular mechanisms participate in the HA interaction with cell membrane of the infected host cell.     

The type II epithelial cells of the lung. III. Lecithin synthesis: a comparison with pulmonary macrophages

Synthesis of lecithins in isolated type II alveolar cells was compared with that in alveolar macrophages as a means of exploring the biochemical mechanisms underlying surfactant production in the lung. Counted cell populations were suspended in a simple glucose-salt solution and 14C-labeled precursors were added singly, in physiologic concentrations, to assess the potential importance of each as a substrate for lecithin synthesis. Molar incorporation of glucose, glycerol, choline, lysolecithin, acetate, palmitate, oleate, and linoleate was determined in lecithins fractionated according to degree of saturation after 1 hour of incubation. Palmitate ws the most actively utilized substrate in type II &cells. Type II cells incorporated 6 nmoles of palmitate per 10(7) cells, of which 77% was in disaturated lecithins, and 66% at the C2 position (compared to 0.8 nmoles, 47% disaturated, in macrophages). Acetate was also incorporated mainly into disaturated lecithins in type II cells; macrophages did not utilize acetate, and no precursor specifically supported disaturated lecithin synthesis in macrophages. Type II cells and macrophages synthesized similar quantities of total lecithins and disaturated lecithins from glucose and choline. Only the type II cells, however, were capable of increasing disaturated lecithin synthesis from 14C-choline when unlabeled palmitate was added to the medium. Type II cells synthesized significantly more disaturated lecithins from lysolecithin than did macrophages (451 versus 60 pmoles per 10(7) cells). Macrophages utilized glycerol in lecithin synthesis, but type II cells did not. Our data demonstrate directly for the first time that type II cells are the site of disaturated lecithin synthesis and that acyl turnover mechanisms are important in production of disaturated lecithins by the type II cell.     

The cardioprotector dexrazoxane augments therapeutic efficacy of mitoxantrone in experimental autoimmune encephalomyelitis

The present study investigates the immunological effects of a combination treatment of mitoxantrone and the cardioprotector dexrazoxane in experimental autoimmune encephalomyelitis (EAE). Mitoxantrone, an anthracycline-derived immunosuppressive drug has been approved recently for treatment of very active multiple sclerosis (MS). Its prolonged use is limited due to its cardiotoxic properties. Dexrazoxane (DZR (S)-(+)-1,2-bis (3,5.dioxopiperazinyl)propane, ICRF-187) is an iron III chelator which in animal models and in cancer patients reduces anthracycline and mitoxantrone induced cardiotoxicity when given immediately before these agents. We examined the immunological effects of dexrazoxane in combination with mitoxantrone in experimental autoimmune encephalomyelitis (EAE) in Lewis rats. EAE was induced by active immunization with myelin basic protein (MBP) or by adoptive transfer of MBP specific T cells (AT-EAE). The clinical course, spinal cord pathology, activity of metalloproteinases (MMP-2 and MMP-9) and T cell apoptosis were assessed. Monotherapy with DZR ameliorated slightly the course of actively induced EAE and AT-EAE. The combination of DZR and mitoxantrone was superior to mitoxantrone given alone. Clinical amelioration ran in parallel with the marked reduction of inflammatory infiltration which was nearly abolished by the combination treatment. DZR did not affect the activity of metalloproteinase 9 and did not increase the proportion of apoptotic lymph node cells ex vivo or T cells in situ. We conclude that in addition to its cardioprotective role, DZR augments mitoxantrone-mediated immunosuppressive effects in animal models of human central nervous system (CNS) autoimmune disease. Clinical trials in MS patients are warranted to evaluate the unexpected immunosuppressive efficacy of DZR as add-on treatment.     

Histamine induced decrease of lecithin levels in broncho-alveolar lavage fluid of rats is mediated by H2 receptor

Lecithin, a major surface active substance of the surfactant system of the lung, was estimated in broncho-alveolar lavage (BAL) fluid in four groups of healthy adult male albino rats. Rats from group I were not administered any drug and acted as controls. Group II were administered histamine diphosphate. Group III were given H1 blocker (pyrilamine maleate) followed by histamine diphosphate. Group IV received H2 blocker (ranitidine hydrochloride) followed by histamine diphosphate. Lecithin content of BAL fluid in the control group was compared with that in the other three groups. A significant decrease in lecithin content was observed in the rats that received either histamine diphosphate or H1 blocker followed by histamine diphosphate. However, compared to control rats no significant difference in lecithin content was seen in rats that received H2 blocker followed by histamine diphosphate. The results clearly indicate that the decrease in surface active lecithin content in BAL fluid following administration of histamine diphosphate was unaffected by prior administration of H1 blocker, but was blocked by prior administration of H2 blocker. It was concluded that histamine induced decrease in lecithin content of BAL fluid is mediated through H2 receptors. Since the predominant source of intra-alveolar lecithin are Type II cells of the alveolar epithelium, It is possible that Type II cells have H2 receptors, stimulation of which resulted in decreased intraalveolar lecithin.     

Low-dosage metronomic chemotherapy and angiogenesis: topoisomerase inhibitors irinotecan and mitoxantrone stimulate VEGF-A-mediated angiogenesis

Metronomic chemotherapy with cytotoxic agents has been shown to inhibit angiogenesis and, consequently, tumor growth by targeting vascular endothelial cells (ECs). In these regimens, anti-tumor activities additional to anti-angiogenesis may operate. Moreover, chemotherapy typically generates reactive oxygen species in targeted ECs, which can affect angiogenesis. The aim of the present study was to assess the systemic effect of low-dosage metronomic treatment with either irinotecan or mitoxantrone on angiogenesis induced by VEGF-A. Angiogenesis was induced in normal adult rat mesentery by intraperitoneal injection of a low dosage of VEGF-A. Thereafter, irinotecan and mitoxantrone were infused separately continuously at minimally toxic dosages for 14 consecutive days via a subcutaneous osmotic minipump. Angiogenesis was assessed in terms of objective and quantitative variables using morphologic and computerized image analyses. Irinotecan or mitoxantrone significantly stimulated angiogenesis, with ironotecan increasing angiogenesis by 104%, when compared with the vehicle-treated animals. Low-dosage metronomic chemotherapy with irinotecan or mitoxantrone stimulates angiogenesis in the normal mesentery of rats, probably by inducing low-level oxidative stress in the targeted ECs. Whether or not this pertains to tumor angiogenesis may be difficult to confirm, as several anti-tumor modes may operate during low-dosage metronomic chemotherapy.     

Chylomicron composition during duodenal triglyceride and lecithin infusion

In lymph fistula rats infused intraduodenally with trioleoylglycerol emulsions, the secretion of triglyceride was enhanced and that of total and esterified cholesterol in chylomicrons was strikingly reduced when lecithin was added at 4-5 times the biliary lecithin secretion rate. During continued triglyceride secretion in the presence of lecithin, the phospholipid: unesterified cholesterol ratio of the chylomicrons increased significantly in contrast to lecithin-free emulsions. Bile-diverted lymph fistula rats secreted significantly less esterified and total cholesterol in lymph chylomicrons than controls, and the difference was magnified by lecithin administration. Cholesterol appears to become relatively unavailable for incorportion into chylomicrons when lecithin is administered at high rates.