注射用精制蜂毒的稳定性研究

应用化学动力学原理考察了本品的稳定性。暂定本品有效期为三年,4℃密闭暗处贮存。     

注射用蜂毒质量考察情况分析

对注射用蜂毒的质量考察,分析结果:室温放置1a后pH值下降0.10±,澄明度微浑至浑浊,含量升高47.81％±。     

异莲心碱抗血小板聚集和抗凝血作用

目的:探讨异莲心碱(isoliensinineI,L)对大鼠体内血小板聚集和凝血功能的影响。方法:以二磷酸腺苷(ADP)诱导血小板聚集,采用比浊法观察IL对兔体内外血小板13,5,min聚集率和最大聚集率的影响,同时评价IL对大鼠凝血酶原时间(PT)、活化部分凝血激酶时间(APTT)及凝血酶时间(TT)的影响。结果:IL 4 mg.L-1和8 mg.L-1能显著抑制ADP诱导的兔体外血小板13,5,min聚集率和最大聚集率,其抑制率为14.4%和27.9%;IL 5 mg.kg-1和10 mg.kg-1能显著抑制ADP诱导的兔体内血小板1,35,min聚集率和最大聚集率,其抑制率为20.0%和32.6%;IL能显著延长大鼠PT、APTT和TT。结论:IL具有对抗血小板聚集和凝血作用。     

可皆霉素的制备及抗凝血作用研究

目的以真菌狭旋毛壳为原料制备可皆霉素,并评价其体外抗凝血活性。方法以狭旋毛壳为原料,从其甲醇提取物中分离得到可皆霉素,并研究其提取工艺。测定可皆霉素对獭兔体外活化部分凝血酶时间(APTT)、凝血酶原时间(PT)、凝血酶时间(TT)和纤维蛋白原(FIB)的影响。结果狭旋毛壳甲醇提取物中可皆霉素的含量高达68.6%,可实现工业化生产。可皆霉素在体外可极显著延长TT(P<0.001),强于阳性对照灯盏花素组(P<0.001),并能显著降低FIB的含量(P<0.001)。结论可皆霉素具有显著的抗凝血活性。     

肝素衍生物的抗凝血活性评价

目的评估一系列化学修饰肝素衍生物的抗凝血活性。方法应用凝血仪测定肝素的部分活化凝血活酶时间(APTT)-肝素浓度的标准曲线,应用酶标仪测定肝素抗Ⅹa因子活性,制定其在405nm的吸光度-肝素浓度标准曲线,然后分别测定肝素衍生物的APTT和抗Ⅹa因子活性,根据标准曲线评价肝素衍生物的抗凝活性。结果化学修饰能够适当降低肝素衍生物的抗凝血活性,并且随着引入的有机基团的增多,肝素衍生物的抗凝血活性逐渐下降,但仍然保持一定的生物活性,且应用APTT评估肝素衍生物抗凝活性与应用抗Ⅹa因子活性评估肝素衍生物抗凝活性,测定结果基本一致。结论相比较肝素,这些化学修饰的肝素衍生物有较低的抗凝血活性,为安全应用这些肝素衍生物提供了更加广阔的前景。     

RAINBOW血凝仪应用时的质量控制

根据国家卫生部出台的停止使用DUKE法测定出血时间、玻片法和毛细血管法测定凝血时间，而由血浆凝血酶原时间(PT)、活化部分凝血酶时间(APTT)和血小板计数联合取代的政策，使原来并非常规检测的PT和APTT成为术前常规检测项目。为此我院引进了德国BE公司的RAINBOW四通道半自动血凝仪。本文主要就该仪器的使用性能及质量控制做如下分析。     

常用凝血指标PT、APTT检测在产前检查中的临床应用

目的通过对96例临产孕妇血浆凝血酶原时间(PT)和活化部分凝血酶原时间(APTT)测定,探讨妊娠晚期妇女凝血功能变化及其在产前检查中的临床意义。方法应用血凝仪对临产孕妇96例、健康非孕育龄妇女90例以凝固法进行检测。结果临产孕妇PT、APTT明显缩短,与正常对照组(非孕育龄妇女)比较差异均有统计学意义(P<0.05)。提示临产孕妇血液中凝血因子处于活化状态。结论临产孕妇的凝血指标较正常非孕育龄妇女有显著差异,机体血液呈高凝状态。凝血指标PT、APTT的产前检查,可早期发现凝血机制的改变,为孕妇的产前防治工作提供科学依据,对预防和治疗产中、产后异常出血和防止DIC发生,提高分娩安全,保护母婴健康具有重要意义。     

地龙抗凝血活性体外评价方法的建立

目的 建立一种基于全自动血凝分析仪测定纤维蛋白原-凝血酶时间的地龙及其相关制剂的抗凝血活性的体外评价方法。方法 以威廉环毛蚓提取物为研究对象,采用BCA蛋白测定法测定可溶性蛋白含量,以抗凝血活性为指标,建立纤维蛋白原-凝血酶时间法测定活性,以活性浓度与可溶性蛋白浓度的比值（比活）作为活性评价指标。结果 10批地龙提取物可溶性蛋白含量为（26.56±3.57）%,比活为（34.64±3.98）U·μg^-1。其比活大小与蚓激酶标准品（43.60 U·μg^-1）相当,且是市售蚯蚓酶（22.17 U·μg^-1）比活的1.5倍。结论 采用BCA蛋白测定法测定地龙提取物可溶性蛋白,方法简便、可靠;所建立的纤维蛋白原-凝血酶时间法,操作简便,人为影响因素小,结果客观准确,可用于实时测定,重复性和仪器精密度良好,线性范围较广,具有可行性,可用于地龙及其制剂活性的评价。     

CA550血凝仪性能评价

目的：对CA550血凝仪的性能进行评价。方法：以凝固法或发色底物法检测凝血酶原时间（PT）、活化部分凝血酶原时间（APTT），同时对它们的精密度、准确度、线性进行分析总结。结果：PT、APTT的批内变异系数（低、中、高）分别为0．6％、0．4％、0．8％、0．5％、1．7％、1．0％；PT、APTT的批间变异系数分别为3．7％，8．9％；PT、APTT线性测定的相关系数分别为0．956、-0．990；PT、APTT携带互污染率分别为-1．3％、-0．6％。结论：CA550血凝仪精密度、准确度，线性都达到了性能要求，因此可进行急诊、批量测定。     

阿藿烯对大鼠抗血栓活性作用研究

目的:研究阿藿烯对大鼠血栓形成和凝血功能的影响,评价其抗血栓活性。方法:将SD大鼠随机分为空白对照组、血塞通阳性药组(50mg/kg)、阿藿烯低剂量组(25mg/kg)、中剂量组(50mg/kg)、高剂量组(75mg/kg),各组动物连续灌胃5d,末次给药2h后建立下腔静脉结扎模型、FeCl3致动脉血栓模型、断尾流血时间测定模型测定阿藿烯对血栓形成和凝血的影响;并测定各组大鼠给药前及给药后2h血浆凝血酶原时间(PT)、活化部分凝血酶时间(APTT)。结果:阿藿烯能抑制动静脉血栓的形成,使血栓重量降低,且高剂量阿霍烯对动静脉血栓形成的抑制率均大于40%;同时有效延长大鼠断尾流血时间,并能使大鼠血浆APTT、PT明显延长(P<0.05),且都具有剂量依赖关系。结论:阿藿烯有抗血栓活性。     

福林-酚法测定蜂毒口服结肠定位释药微丸含量

目的建立天然蜂毒多肽口服结肠定位释药微丸的含量测定方法。方法采用福林 酚试剂法。结果蜂毒多肽质量浓度在 2 5～ 2 5 0mg/L内线性关系良好 ,相关系数为 0 9994。测定微丸样品的平均回收率为 1 0 0 5 % ,平均相对标准差为 1 1 % (n =9)。结论此方法可以满足制剂样品含量分析的要求     

蜂毒诱发大鼠结肠炎疼痛实验模型的建立

目的建立蜂毒诱发大鼠结肠炎疼痛实验模型,用于镇痛药的评价及相关机理研究。方法利用蜂毒结肠粘膜下注射诱发大鼠结肠炎疼痛,动态定量评价大鼠的疼痛行为学反应,并进行组织学检查,同时与甲醛相比较。结果每只大鼠结肠粘膜下注射蜂毒0.075,0.15和0.3mg可剂量依赖性地诱发大鼠持续性内脏痛,与5%甲醛(0.1mL)相比,蜂毒0.3mg致痛反应效应相当,第二相炎症反应明显,且持续期长;局部组织损伤轻。阿片类镇痛药吗啡、解热镇痛药阿司匹林和新型环氧合酶2抑制剂NS-398均能抑制蜂毒诱发的大鼠结肠炎疼痛。结论蜂毒诱发大鼠结肠炎疼痛模型反应指标客观、实验重复性好,是一种较理想的内脏疼痛实验模型,可用于镇痛药的评价及机理研究。     

蜂毒多肽空间稳定免疫脂质体的制备及体外对肿瘤细胞的选择性

蜂毒多肽在肿瘤治疗中的应用引起研究者的极大兴趣。本研究使用大豆磷脂、胆固醇、羧酸化PEG-胆固醇制备了蜂毒多肽空间稳定脂质体，并将二硫键稳定抗人肝癌单链抗体联结PEG-胆固醇末端。使用酶联免疫法考察了蜂毒多肽空间稳定免疫脂质体的活性。蜂毒多肽空间稳定免疫脂质体有较高的肿瘤细胞选择性。体外实验证明，其对SMMC-7721细胞的杀伤能力远强于蜂毒多肽空间脂质体，而对Hela细胞的杀伤能力与蜂毒多肽空间脂质体无区别。蜂毒多肽空间稳定免疫脂质体对肿瘤细胞的选择性，可使其成为一种有效的靶向制剂。     

中华眼镜蛇毒组分F的体外抗血管生成活性研究

目的研究中华眼镜蛇毒组分F对血管内皮细胞黏附功能的抑制作用和体外的抗血管生成活性。方法MTT快速测定法测定组分F对原代培养的血管内皮细胞对纤维蛋白原(Fg)、纤连蛋白(Fn)、Matrigel的黏附作用的影响;采用平面拟毛细血管生成培养法,观察组分F对血管生成的抑制效应。结果组分F可抑制血管内皮细胞的黏附功能,对细胞黏附Fn、Fg和Matrigel 3种物质的作用从1.2μg/ml浓度起即有抑制效应(P<0.05),且均呈浓度依赖性,组分F对细胞黏附Fg和Matrigel的抑制作用较其对Fn的作用强(P<0.05);组分F可抑制内皮细胞在培养基质中生成血管样网状结构的反应,并且随浓度的不同抑制程度有明显不同。结论中华眼镜蛇毒组分F具有体外抗血管生成活性。     

蜂毒的主要成分及药理作用的研究进展

蜂毒是由蜜蜂毒腺分泌的活性物质,其中很多活性成分具有抗炎、抗肿瘤、镇痛、降压的作用。近年来,国内外很多科学家对蜂毒的活性成分及其基因结构、药理作用机制、分子生物学等方面进行了研究,并且取得了重要的研究进展。本文总结了蜂毒的主要活性成分以及主要的药理作用,为蜂毒的临床应用提供理论基础。     

Combined antitumor effects of bee venom and cisplatin on human cervical and laryngeal carcinoma cells and their drug resistant sublines

In the present study, we investigated the possible combined anticancer ability of bee venom (BV) and cisplatin towards two pairs of tumour cell lines: parental cervical carcinoma HeLa cells and their cisplatin‐resistant HeLa CK subline, as well as laryngeal carcinoma HEp‐2 cells and their cisplatin‐resistant CK2 subline. Additionally, we identified several peptides of BV in the BV sample used in the course of the study and determined the exact concentration of MEL. BV applied alone in concentrations of 30 to 60 µg ml^–1 displayed dose‐dependent cytotoxicity against all cell lines tested. Cisplatin‐resistant cervical carcinoma cells were more sensitive to BV than their parental cell lines (IC₅₀ values were 52.50 µg ml^–1 for HeLa vs. 47.64 µg ml^–1 for HeLa CK cells), whereas opposite results were obtained for cisplatin‐resistant laryngeal carcinoma cells (IC₅₀ values were 51.98 µg ml^–1 for HEp‐2 vs. > 60.00 µg ml^–1 for CK2 cells). Treatment with BV alone induced a necrotic type of cell death, as shown by characteristic morphological features, fast staining with ethidium‐bromide and a lack of cleavage of apoptotic marker poly (ADP‐ribose) polymerase (PARP) on Western blot. Combined treatment of BV and cisplatin induced an additive and/or weak synergistic effect towards tested cell lines, suggesting that BV could enhance the killing effect of selected cells when combined with cisplatin. Therefore, a greater anticancer effect could be triggered if BV was used in the course of chemotherapy. Our results suggest that combined treatment with BV could be useful from the point of minimizing the cisplatin concentration during chemotherapy, consequently reducing and/or postponing the development of cisplatin resistance. Copyright © 2013 John Wiley & Sons, Ltd.     

Inhibition of mammary carcinoma cell proliferation in vitro and tumor growth in vivo by bee venom.

The possible tumor growth- and metastasis-inhibiting effects of bee venom in mice and in tumor cell cultures were studied. The tumor was a transplantable mammary carcinoma (MCa) of CBA mouse. Intravenous administration of bee venom to mice significantly reduced the number of metastases in the lung. However, subcutaneous administration of bee venom did not reduce the number of lung metastases, indicating that the antitumor effect of the venom could be highly dependent on the route of injection as well as close contact between the components of the venom and the tumor cells, as was shown by in vitro studies on MCa cells. We also observed variations in immunological parameter induced by bee venom. We proposed that bee venom has an indirect mechanism of tumor growth inhibition and promotion of tumor rejection that is based on stimulation of the local cellular immune responses in lymph nodes. Apoptosis, necrosis, and lysis of tumor cells are other possible mechanisms by which bee venom inhibits tumor growth.     

Pharmacological properties and therapeutic potential of honey bee venom

Honey bee venom (BV) is a valuable product, and has a wide range of biological effects, and its use is rapidly increasing in apitherapy. Therefore, the current study, we reviewed the existing knowledge about BV composition and its numerous pharmacological properties for future research and use. Honey bee venom or apitoxin is produced in the venom gland in the honey bee abdomen. Adult bees use it as a primary colony defense mechanism. It is composed of many biologically active substances including peptides, enzymes, amines, amino acids, phospholipids, minerals, carbohydrates as well as some volatile components. Melittin and phospholipase A₂ are the most important components of BV, having anti-cancer, antimicrobial, anti-inflammatory, anti-arthritis, anti-nociceptive and other curative potentials. Therefore, in medicine, BV has been used for centuries against different diseases like arthritis, rheumatism, back pain, and various inflammatory infections. Nowadays, BV or its components separately, are used for the treatment of various diseases in different countries as a natural medicine with limited side effects. Consequently, scientists as well as several pharmaceutical companies are trying to get a new understanding about BV, its substances and its activity for more effective use of this natural remedy in modern medicine.     

Characteristics of Adverse Events in Bee Venom Therapy Reported in South Korea: A Survey Study

This study was aimed at investigating Korean patients’ experience with bee venom therapy (BVT) and providing evidence to enhance BVT safety. Thus, an anonymous online survey was conducted between August 22 and 28, 2018. Five hundred respondents who underwent pharmacopuncture (PA) within one year were surveyed (sample error: 95 ± 4.38%). Of these, 32 respondents were excluded and 468 were evaluated. Of the 468, 61 reported experiencing adverse events after PA. The adverse event rate was higher in the BV-PA(Bee venom-Pharmacopuncture) group than in the non-A group; however, intergroup differences were insignificant. There were no significant differences in mild symptom intensity between the BV-PA and non-BV-PA groups (p = 0.572). However, there was a significant intergroup difference in severe symptom intensity (p < 0.001). Additionally, the BV-PA and non-BV-PA groups did not significantly differ in their level of satisfaction either overall or in terms of effectiveness and safety (p = 0.414, p = 0.339, and p = 0.675, respectively). Furthermore, the BV-PA and non-BV-PA groups did not differ regarding intent to re-treat (p = 0.722). Severe adverse events such as anaphylactic shock were not reported; however, BVT practitioners should be cautious when applying it.