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）。     

柴平左金汤治疗胆汁反流性胃炎58例

[目的]观察柴平左金汤治疗胆汁反流性胃炎疗效。[方法]将116例随机分为两组。治疗组58例采用自拟柴平左金汤,对照组58例应用雷尼替丁、吗丁啉口服。治疗60d疗效判定。[结果]治疗组痊愈26例,显效16例,有效12例,无效4例,总有效率93.10%。对照组痊愈5例,显效10例,有效26例,无效17例,总有效率70.69%。治疗组疗效优于对照组(P<0.01)。[结论]柴平左金汤治疗胆汁反流性胃炎具有较好疗效。     

LC-MS/MS同时测定左金丸和香连丸中6种生物碱的含量

目的:建立左金丸(黄连、吴茱萸)和香连丸(黄连(吴茱萸制)、木香)中6种生物碱含量的LC-MS/MS测定方法。方法:色谱条件:Phenomenex Luna C18柱(5μm,250 mm×4.60 mm);乙腈-水(4 mmol/L乙酸铵和0.08%甲酸)为流动相,梯度洗脱;质谱条件:采用电喷雾离子源(ESI),正离子检测,MRM模式。结果:左金丸中药根碱、黄连碱、巴马汀、小檗碱、吴茱萸碱和吴茱萸次碱的平均回收率分别为101.0%,101.3%、101.2%、100.2%、99.6%、99.7%,RSD分别为1.1%、1.8%、0.98%、1.0%、1.3%、1.3%,香连丸中药根碱、黄连碱、巴马汀、小檗碱和吴茱萸碱的平均回收率分别为101.6%、99.8%、101.9%、101.0%、101.3%,RSD分别为1.0%、1.9%、0.75%、1.1%、1.9%。结论:本方法可同时测定左金丸和香连丸中药根碱、黄连碱、巴马汀、小檗碱、吴茱萸碱和吴茱萸次碱的含量,具有快速、简便等特点。     

左金丸抗幽门螺杆菌感染的分子网络调控机制研究

目的基于网络药理学和生物信息学方法探究左金丸抗幽门螺杆菌(Hp)感染的分子网络调控机制。方法通过中药系统药理学数据库和分析平台(TCMSP)检索左金丸化学成分,筛选并预测其入血活性成分和作用靶点,检索疾病数据库中与Hp感染相关的作用靶点,构建左金丸成分靶点与疾病靶点的交互网络,获得左金丸抗Hp的特征性基因;通过DAVID6.8数据库对上述特征性基因进行GO功能富集和KEGG通路富集分析;利用Cytohubba筛选出左金丸抗Hp感染的关键靶点。结果通过TCMSP筛选、预测得到左金丸32个入血活性成分和197个作用靶点。GO分析共得到199条富集结果,其中生物过程包含炎症反应、RNA信号转录、信号传导等152条,细胞组分包含细胞核、细胞外基质、蛋白复合物等19条,分子功能包含细胞因子活性、DNA结合、ATP结合等28条。KEGG分析结果显示,Jak-STAT信号通路、T细胞受体信号通路、细胞周期信号通路、Wnt信号通路等65条通路与左金丸抗Hp感染密切相关。经Cytohubba筛选得到CXCL8、IL10、IL4、VEGFA、MMP9等10个关键基因。结论左金丸抗Hp感染具有多成分、多靶点、多通路协同作用的特点,可为其活性成分研究和抗Hp药效及机制研究提供依据。     

5-氨基水杨酸pH依赖型控释微丸的制备及质量评价

目的:制备5-氨基水杨酸(美沙拉嗪)pH依赖型控释微丸,考察影响载药丸芯制备和微丸控释的因素.方法:以圆整度、粒度分布和收率、脆碎度和流动性为指标,采用正交设计法筛选丸芯的处方和工艺以及包衣的增重和老化条件.结果:最佳处方工艺为载药量80％、粘合剂用量40mL/100 g、挤出速度30 Hz、滚圆速度30 r·min-1;包衣增重8％,老化条件为60℃、24 h.结论:制备了适合流化床包衣的美沙拉嗪载药丸芯,以尤特奇水分散体包衣得到了pH依赖型控释微丸.     

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。     

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.     

左金方对幽门螺旋杆菌感染的人GES-1细胞增殖与凋亡的作用研究

目的研究左金方对幽门螺旋杆菌引起人正常胃黏膜上皮(GES-1)细胞增殖与凋亡的影响。方法幽门螺旋杆菌以不同的感染复数(1∶1、50∶1、100∶l、200∶1、300∶1)感染GES-1细胞,于12、24、48 h后,CCK-8检测细胞的增殖活性;以不同质量浓度(0.5、1.0、2.0、4.0μg/m L)左金方作用幽门螺旋杆菌感染的GES-1细胞12、24、48 h后,收集细胞,CCK-8检测细胞的增殖活性,流式细胞仪和Western blotting检测细胞的凋亡率;Hochest染色检测凋亡细胞的形态学改变。结果幽门螺旋杆菌感染GES-1细胞后,感染复数越大,侵染时间越长,抑制细胞生长和促进细胞凋亡的作用越强,幽门螺旋杆菌以100∶1的感染复数作用人GES-1细胞12、24、48 h后,细胞存活率分别降低到(80.57±1.21)%、(70.04±3.21)%、(67.74±2.91)%,细胞的凋亡率分别增加到(23.74±1.71)%、(53.60±1.87)%、(70.67±2.87)%;与1.0μg/m L左金方共培养24 h后,细胞存活率上升到(97.67±1.04)%,细胞凋亡率降低到(31.04±1.02)%,与相应模型组对比,差异显著(P0.01);Westernblotting结果表明,左金方各处理组细胞Caspase-3蛋白水平表达均明显降低;Hochest荧光染色,从形态学的角度进一步验证了这一结果。结论幽门螺旋杆菌感染人GES-1细胞后,可以抑制细胞增殖,诱导细胞凋亡,左金方可以降低幽门螺旋杆菌引起的细胞损伤,保护GES-1细胞。     

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。