小鼠扩张囊胚颗粒法玻璃化冷冻保存技术研究

在２５℃室温下,用ＥＦＳ３０玻璃化溶液,对小鼠扩张囊胚进行颗粒法玻璃化冷冻保存。结果,胚胎在ＥＦＳ３０溶液中平衡１ｍｉｎ后滴入液氮中,解冻后的胚胎发育率达１００％,囊胚孵化率达７６％。而在１０％乙二醇溶液中将胚胎预处理５ｍｉｎ,再移入ＥＦＳ３０中平衡１ｍｉｎ后冷冻保存的胚胎,解冻后的发育率和囊胚孵化率分别为９８％和６３％。上述２组的胚胎发育率与对照组无明显差异,而囊胚发育率均明显低于对照组（Ｐ＜０．０１）。上述２组胚胎解冻后植入假孕小鼠子宫后的受体妊娠率分别为９０％和８１％,产出率分别为７１％和５８％,与对照组（６１％）相比均无显著性差异（Ｐ＞０．０５）。     

Influence of MCC II fraction and storage conditions on pellet properties

Microcrystalline cellulose II (MCC II) – a polymorph of commonly used MCC I – was introduced as new pelletization aid in wet-extrusion/spheronization leading to fast disintegrating pellets. Previous investigations suggested that pellet properties were influenced by the fraction of MCC II. Furthermore, it is unknown whether the storage conditions can affect the disintegration behavior. Therefore, the effects of MCC II fraction and the storage conditions on several pellet properties were investigated.MCC II-based pellets were prepared of pure MCC II or binary mixtures containing 10–50% (steps of 10%) MCC II as pelletization aid and theophylline, chloramphenicol or lactose. The pellets were characterized by their aspect ratio, equivalent diameter, water content, tensile strength, porosity as well as shrinking, and disintegration behavior and drug release according to their MCC II fraction. Furthermore, the pellets were stored at different relative humidities (0–97%rh), and the influence on their disintegration and drug release was investigated.With increasing MCC II fraction, the pellets became lager in size, decreased their porosity, and required higher water contents for spheronization. Moreover, the disintegration time increased and the disintegration itself was incomplete. Furthermore, the storage conditions had an impact on the disintegration properties of MCC II-based pellets. The disintegrating was affected irreversibly after storage at high humidity (80–97%rh) resulting in a slow drug release. Therefore, MCC II-based pellets need to be stored below 80%rh to secure a fast disintegration.A better knowledge of the properties of MCC II-based pellets was obtained providing a basis for a successful manufacturing and adequate storage of MCC II-based pellets prepared by extrusion/spheronization.     

Construction of a Tissue‐Engineered Annulus Fibrosus

The intervertebral disc is composed of load‐bearing fibrocartilage that may be subjected to compressive forces up to 10 times the body weight. The multilaminated outer layer, the annulus fibrosus (AF), is vulnerable to damage and its regenerative potential is limited, sometimes leading to nuclear herniation. Scaffold‐based tissue engineering of AF using stem cell technology has enabled the development of bi‐laminate constructs after 10 weeks of culture. It is difficult to know if these constructs are limited by the differentiation state of the stem cells or the culture system. In this study, we have characterized an expandable scaffold‐free neoconstruct using autologous AF cells. The construct was prepared from pellet cultures derived from monolayer cultures of AF cells from mature pigs that became embedded in their own extracellular matrix. The pellet cultures were incubated for 24 h in a standardized conical tube and then carefully transferred intact to a culture flask and incubated for 21 days to allow continued matrix synthesis. Cell viability was maintained above 90% throughout the culture period. The engineered scaffold‐free construct was compared with the native AF tissue by characterization of gene expression of representative markers, histological architecture, and biochemical composition. The morphological and biochemical characteristics of the cultured disc construct are very similar to that of native AF. The cell number per gram of construct was equal to that of native AF. Expression of aggrecan was elevated in the engineered construct compared with RNA extracted from the AF. The glycosaminoglycan content in the engineered construct showed no significant difference to that from native construct. These data indicate that scaffold‐free tissue constructs prepared from AF cells using a pellet‐culture format may be useful for in vitro expansion for transplantation into damaged discs.     

不同处方复方丹参黏附微丸黏附性能的评价

目的考察不同处方复方丹参黏附微丸的黏附性能,从而筛选出黏附性能较为合适的处方。方法以壳聚糖、羟丙基甲基纤维素K100M(HPMCK100M)、卡波姆为黏附材料,制备不同处方的复方丹参黏附微丸,并对其黏附性能进行测定。体外黏附性能的测定采用了最小剥离力法即测定黏附微丸与猪小肠的剥离力,组织残留法即测定微丸在猪小肠的滞留数。体内黏附性能的测定采用了直接灌胃法,通过测定微丸在小肠各部位的滞留数目,计算其黏附指数。结果组织残留法试验结果在相同时间内黏附材料占比40%的微丸残留数目较黏附材料占比30%的微丸多,最小剥离力法试验结果表明黏附材料占比40%的微丸其最小剥离力较大,同组织残留法结果一致。体内黏附试验显示黏附材料占比40%的微丸其黏附指数较小,但在胃中有一定的滞留。结论当黏附材料总占比一致时,各处方之间黏附性能并没有显著性差异,黏附材料增加时其黏附性能也会增加,但会增加在胃中的滞留时间,可能会破坏方中的有效成分。     

Utilizing mixer torque rheometer in the prediction of optimal wet massing parameters for pellet formulation by extrusion/spheronization

In this study, we aimed to optimize theophylline pellet formulations using a two-factor three-level full-factorial design (3²) by monitoring the concentration of two pellet excipients, polyvinyl pyrrolidone K30 (PVP) binder solution (X1) and the hydrophilic excipient mannitol (X2). Their impact on pellet characteristics (responses) were evaluated. Increasing PVP concentration in the binder solution resulted in an increase in the wet mass torque value. The effect of mannitol, however, was antagonistic. Moreover, the pellet particle size was significantly influenced by the level of mannitol, PVP solution, and quadratic effect of mannitol. Mannitol significantly antagonized the pellet particle size. Furthermore, increased mannitol concentrations significantly enhanced drug dissolution rate from the pellets, whereas PVP concentration in the binder solution significantly reduced the drug dissolution rate. In conclusion, wet granulations can be controlled by monitoring the composition of the binder solution and pellet composition.     

Penetrating Facial Injury with an “Airsoft” Pellet: A Case Report

Airsoft is a recreational combat sport that originated in Japan in the 1970s and is currently increasing in popularity in the UK. Participants use air or electrically powered weapons to fire small plastic pellets at a controlled pressure. UK law strictly regulates the maximum muzzle velocity and the type of ammunition used in these weapons. A search of published papers found several reports of penetrating ocular injuries caused by Airsoft pellets, but no reports of penetrating injuries to other areas of the body. We report the case of a 25-year-old man who sustained a penetrating injury to the cheek after being shot with an Airsoft weapon.     

大黄(庶虫)虫丸对实验性子宫内膜异位症大鼠免疫功能的影响

目的:观察大黄(庶虫)虫丸对EMT大鼠免疫功能的影响.方法:采用大鼠子宫内膜异位症(EMT)动物模型,造模成功后按照异位内膜体积将大鼠随机分为大黄(庶虫)虫丸中高剂量组、大黄(庶虫)虫丸中低剂量组、西药组、模型组,并以假手术组为对照,灌胃治疗4周后检测指标.结果:①大黄(庶虫)虫丸中高剂量组、大黄(庶虫)虫丸中低剂量组、西药组对异位子宫内膜有明显抑制作用,与治疗前异位内膜体积(V1)相比,异位内膜体积(V2)明显缩小(P<0.01);②大黄(庶虫)虫丸中高剂量组能明显提高EMT大鼠胸腺、脾脏重量指数(P<0.01),降低血清肿瘤坏死因子-α(TNF-α)水平(P<0.01);③病理组织学分析显示:大黄(庶虫)虫丸中高剂量组能明显增加胸腺皮质厚度、脾小节大小及淋巴细胞数,明显降低异位内膜腺体体密度、提高腺腔值、降低淋巴结生发中心体密度(P<0.01).结论:大黄(庶虫)虫丸治疗子宫内膜异位症的部分机制可能改善子宫内膜异位症大鼠免疫功能有关.     

紫草止痒丸止痒抗炎作用的药效学研究

目的：观察紫草止痒丸的止痒、抗炎作用。方法：磷酸组织胺致痒法，二甲苯致小鼠耳廓肿胀法。结果：紫草止痒丸能明显提高豚鼠致痒阈及抑制二甲苯所致的小鼠耳廓肿胀。结论：紫草止痒丸具有止痒与抗炎作用。     

左金胃漂浮缓释片中小檗碱、巴马汀、吴茱萸碱和吴茱萸次碱的体外释放规律和机制研究

目的探讨左金胃漂浮缓释片体外多成分释放的规律和机制。方法采用转篮法以人工胃液为介质,以HPLC法测定左金胃漂浮缓释片中小檗碱、巴马汀、吴茱萸碱和吴茱萸次碱在8 h内的体外累积释放率,通过相似因子比较法、Higuchi方程,零级、一级释放方程等释放模型拟合法以及Peppas方程研究左金胃漂浮缓释片多成分释放的规律。结果左金胃漂浮缓释片中4种生物碱的平均释放曲线相互之间的相似因子均大于80%,均以Higuchi释放模型为最佳拟合模型,释放机制均为扩散协同骨架溶蚀作用。结论左金胃漂浮缓释片中生物碱类成分的释放具有均衡缓释性。     

龟鹿补肾丸和龟鹿补肾口服液对雄性生殖系统影响的研究

龟鹿补肾丸和龟鹿补肾液明显增加未成熟的小白鼠和去势大白鼠附性器官的重量;龟鹿补肾液能明显增强正常雄性和去势大白鼠的交配能力,随着剂量的加大,能提高未成熟大白鼠血清睾酮水平,而龟鹿补肾丸的影响不如龟鹿补肾液。