重组人胰岛素样生长因子-1分离纯化工艺的建立

目的：建立重组人胰岛素生长因子－1（rhIGF-1)大肠杆菌表达后的高效分离纯化工艺，以便获得高纯度的重组蛋白。方法：已构建好的表达载体转化大肠杆菌BL21，经IPTG诱导进行表达，将离心收集到的菌体用超声破壁，破壁菌液离心分离，收集上清液，进行阴离子交换层析，疏水层析和分子筛纯化，得到最终纯品，蛋白纯度用SDS－PAGE鉴定，ELISA法检测rhIGF-1含量，检测纯化后重组蛋白的生物学活性。结果：用大肠杆菌BL21表达rhIGF-1以可溶性形式存在胞浆中，经3步rhIGF-1分离纯化工艺，其工艺流程可获得具有生物学活性的高纯度rhIGF-1，为产业化及临床应用奠定基础。     

植物多糖纯化分离方法研究进展

多糖具有重要的生物活性,分离纯化过程是影响多糖活性的关键环节。对近年来植物多糖的分离纯化方法的研究进展进行了综述,分析其原理及其优缺点,以期为多糖的研究开发和工业化生产提供参考依据。     

植物多糖提取分离及药理作用的研究进展

对近年来植物多糖在提取、分离纯化和药理作用等方面的发展做一综述，为以后植物多糖的研究奠定了基础。     

多糖的提取分离及纯化研究

对多糖的认识备受关注,常用的提取方法有:溶剂提取法、酸提取法、碱提取法、酶解法、超滤、超声波、微波、超临界流体萃取;分离纯化技术有:沉淀法、柱层析.本文对以上种实验方法做一简单介绍.     

永川豆豉胰蛋白酶抑制剂的分离纯化及其降糖活性研究

目的从豆豉中提取胰蛋白酶抑制剂,并探讨其降血糖活性。方法以永川豆豉为材料,经脱脂,酸性溶液提取,硫酸铵沉淀得到胰蛋白酶抑制剂粗提物,再经Sephadex G-75凝胶层析得到纯化的胰蛋白酶抑制剂,以纯化的胰蛋白酶抑制剂给糖尿病小鼠灌胃并观察其降血糖活性。结果从豆豉里分离出的胰蛋白酶抑制剂对胰蛋白酶的抑制率为30%~40%;用抑制剂提取液给四氧嘧啶糖尿病模型小鼠连续灌胃4 d,发现其血糖值明显低于模型组;观察小鼠的胰组织病理切片,与糖尿病小鼠比较,灌胃后的小鼠胰组织明显得到修复。结论豆豉提取液有较明显的降糖作用。     

虎杖多糖的分离纯化及结构研究

目的分离纯化虎杖总多糖,并对其结构进行分析。方法采用水提醇沉法提取虎杖总多糖,联合DEAE-纤维素、Sephadex G-100柱色谱对虎杖总多糖进行分离纯化;采用高效凝胶渗透色谱法对均一多糖进行纯度鉴定和分子量测定;PMP柱前衍生-高效液相色谱法(HPLC)测定单糖组成;甲基化、气质联用(GC-MS)、红外光谱和核磁共振波谱对其结构进行分析。结果经分离纯化得到2个均一多糖PPⅠ、PPⅡ,其相对分子量分别为7780 Da、5720 Da。单糖组成实验表明PPⅠ、PPⅡ主要由葡萄糖构成。甲基化实验表明PPⅠ、PPⅡ均以1→4、1→3,4、1→4,6葡萄糖苷键存在,摩尔比分别为17∶1∶4、21∶1∶3。红外光谱及核磁波共振波谱对其结构分析证明PPⅠ、PPⅡ由α-型吡喃糖连接而成。结论 PPⅠ、PPⅡ是由α-(1,4)、α-(1→3,4)α-(1→4,6)吡喃葡萄糖苷键连接而成的不同结构、不同分子量的均一多糖。为进一步研究虎杖均一多糖的结构及活性提供依据。     

中药蜈蚣多糖的分离纯化及性质研究

目的:从中药蜈蚣体内提取多糖组分,并对其部分性质进行分析.方法:从中药蜈蚣体内提取粗多糖,经DEAE-52离子交换柱层析分离获得3个糖组分;凝胶过滤法测组分Ⅰ的分子量;MTT法检测多糖组分Ⅰ对两种肿瘤细胞的抑制作用.结果:组分Ⅰ的分子量为33.1 kD;该多糖组分对Hela细胞的抑制作用明显,当糖浓度为3.13 μg/mL时,抑制率达60.8%,但对Eca 109细胞无明显抑制作用.结论:蜈蚣多糖组分Ⅰ对不同的肿瘤细胞具有特异性.     

土鳖虫多肽的分离纯化及溶栓活性研究

目的 对土鳖虫多肽进行分离纯化,并对其溶栓活性进行研究.方法 采用仿生酶解工艺制备土鳖虫复合多肽,以溶栓活性为指标,采用Sephadex G-25、SP Sephadex C-25凝胶柱和RP-HPLC C18半制备色谱柱对复合多肽进行分离纯化,并测定目标多肽组分(Peak b)的溶栓活性、肽含量和分子量.结果 目标多肽组分的溶栓活性为814 430 U·mg-1,肽含量为95.7％,分子量分布在3 211 ～3 547.结论 利用仿生酶解制备的土鳖虫多肽,经分离纯化后,得到一具有较强溶栓活性的多肽组分.     

中药有效成分的分离纯化技术研究概况

文章总结了中药有效成分分离纯化的目的和方法。重点阐述了超速离心法、凝胶层析法、膜分离技术、大孔吸附树脂分离纯化、高速逆流色谱技术、分子蒸馏技术等应用概况。     

白藜芦醇分离纯化及检测技术研究进展

本文主要针对近年来有关白藜芦醇提取、纯化和定量检测技术的研究进展进行了综述。其中粗提技术包括有机溶剂浸提、碱提法、酶法、超临界CO2萃取、微波以及超声波辅助提取等;纯化技术有柱层析、高速逆流色谱、大孔吸附树脂等;检测技术有高效液相色谱、毛细管电泳、飞行时间质谱、气相色谱、色谱联用技术等。     

苦瓜子中植物胰岛素的凝胶柱与RP-HPLC法分离

采用有机酸、醇提取,Sephadex G-50凝胶柱层析、反相高效液相色谱(RP-HPLC)技术分离,从苦瓜子中分离出降糖多肽PA,根据SDS-PAGE电泳和氨基酸组成分析,判定PA为植物胰岛素.     

金芪降糖片联合胰岛素治疗胰岛素抵抗患者低血糖发作临床研究

目的：探讨金芪降糖片联合胰岛素治疗胰岛素抵抗患者低血糖发作的临床疗效。方法：对2011年1月至2011年12月间收治的100例胰岛素抵抗的糖尿病患者进行回顾性分析,按所采用治疗方法分为金芪降糖片联合胰岛素泵组（联合组）与单纯胰岛素泵组（单药组）,每组各50例。结果：联合组患者的HbAlC、FINS、胰岛素分泌指数、2hPBG、MAGE及治疗结束时胰岛素用量（胰岛素／体质量）分别是（5．7±0．7）％、（1．6±0．3）mlU·L^-1、（0．27±0．04）mmol·L^-1、（8．6±1．9）mmol·L^-1、（2．8±0．6）mmol·L^-1、（0．14±0．03）u·kg^-1,均小于单药纽（6．3±1．1）％、（2．1±0．6）mlU·L^-1、（0．31±0．05）mmol·L^-1、（9．4±2．1）mmol·L^-1、（3．1±0．8）mmol·L^-1、（0．17±0．05）u·kg^-1,与单药组比较,差异均有统计学意义（P〈0．05）;FBG与MBG在联合组中虽与单药组差异无统计学意义（P〉0．05）,但与治疗前比较有所改善。治疗期间,联合组患者共发作低血糖4例,单药组出现13例,两组比较,差异有统计学意义（P〈0．05）。结论：对于胰岛素抵抗的糖尿病患者,联合运用金芪降糖片可以减少胰岛素用量,有效改善血糖的波动,减少低血糖的发作。     

点穴膏的主要药效学研究

点穴膏系中药复方外用抗癌经验方，研究结果表明，其对小鼠Ｓ１８０肉瘤、小鼠艾氏腹小癌实体及大鼠Ｗ２５６均有明显的抑制作用，并对醋酸刺激、热刺激以及电刺激的小鼠或大鼠有明显的镇痛作用。对豚鼠皮肤的刺激性属轻度刺激性，从而为点穴膏抗癌作用及治疗癌性疼痛提供了重要的药理学依据。     

黄芪葛根汤对实验性糖尿病及胰岛素抵抗的影响

目的:观察黄芪葛根汤对实验性糖尿病及胰岛素抵抗(IR)的影响。方法:建立链脲佐菌素(STZ)ip所致糖尿病小鼠模型,sc氢化可的松琥珀酸钠(HCSS)诱导的IR小鼠模型,以及采用给大鼠1次ip小剂量链脲佐菌素(STZ),并加饲高热量饮食(富含脂肪和蔗糖),制备2型糖尿病(T2DM)伴胰岛素抵抗(IR)大鼠模型,观察黄芪葛根汤对糖脂代谢以及IR的影响。结果:黄芪葛根汤降低糖尿病小鼠的空腹血糖(FBG),提高糖耐量(OGTT),改善小鼠对胰岛素的敏感性。黄芪葛根汤能显著降低2型糖尿病胰岛素抵抗大鼠的FBG,提高OGTT,降低高胰岛素水平,提高胰岛素耐量,增加胰岛素敏感指数(ISI)和降低IR指数(HOMA-IR)指数。同时,黄芪葛根汤还可调节血脂、下调血清中游离脂肪酸(FFA)的水平。结论:黄芪葛根汤通过改善糖脂代谢、提高胰岛素的敏感性,发挥其治疗糖尿病及胰岛素抵抗的作用。     

冬连胶囊对实验性糖尿病大鼠早期胰岛素抵抗的影响

Objective To investigate the effects of Donglian Capsule(the combination of Ophiopogonis Tuber Polysaccharide with Rhizoma Coptidis Alkaloids, OPRA for short, Component of Chinese medicine)on early insulin resistance in experimental diabetic rats. Methods The animal model of insulin resistance in 2 type diabetic was established by injecting low dose of STZ in 60 high fat-fed SD rats. After one week, the successful models were randomly divided into 4 groups: untreated, high and low OPRA-treated, mefformin-treated group, Meanwhile, a normal control group was designed. After four weeks, the differences of FBG, PG, INS and C-P were compared among the groups. Results The OPRA can significantly reduced the level of serum TC and FBG, increased the C-E ]Lightened the IRI(P<0.01). Conclusion The OPRA can reduce the level of serum TG and FBG, and decrease insulin resistance to protect the β-cell function.     

冬连胶囊对实验性糖尿病大鼠早期胰岛素抵抗的影响

Objective To investigate the effects of Donglian Capsule(the combination of Ophiopogonis Tuber Polysaccharide with Rhizoma Coptidis Alkaloids, OPRA for short, Component of Chinese medicine)on early insulin resistance in experimental diabetic rats. Methods The animal model of insulin resistance in 2 type diabetic was established by injecting low dose of STZ in 60 high fat-fed SD rats. After one week, the successful models were randomly divided into 4 groups: untreated, high and low OPRA-treated, mefformin-treated group, Meanwhile, a normal control group was designed. After four weeks, the differences of FBG, PG, INS and C-P were compared among the groups. Results The OPRA can significantly reduced the level of serum TC and FBG, increased the C-E ]Lightened the IRI(P<0.01). Conclusion The OPRA can reduce the level of serum TG and FBG, and decrease insulin resistance to protect the β-cell function.     

冬连胶囊对实验性糖尿病大鼠早期胰岛素抵抗的影响

Objective To investigate the effects of Donglian Capsule(the combination of Ophiopogonis Tuber Polysaccharide with Rhizoma Coptidis Alkaloids, OPRA for short, Component of Chinese medicine)on early insulin resistance in experimental diabetic rats. Methods The animal model of insulin resistance in 2 type diabetic was established by injecting low dose of STZ in 60 high fat-fed SD rats. After one week, the successful models were randomly divided into 4 groups: untreated, high and low OPRA-treated, mefformin-treated group, Meanwhile, a normal control group was designed. After four weeks, the differences of FBG, PG, INS and C-P were compared among the groups. Results The OPRA can significantly reduced the level of serum TC and FBG, increased the C-E ]Lightened the IRI(P<0.01). Conclusion The OPRA can reduce the level of serum TG and FBG, and decrease insulin resistance to protect the β-cell function.     

冬连胶囊对实验性糖尿病大鼠早期胰岛素抵抗的影响

Objective To investigate the effects of Donglian Capsule(the combination of Ophiopogonis Tuber Polysaccharide with Rhizoma Coptidis Alkaloids, OPRA for short, Component of Chinese medicine)on early insulin resistance in experimental diabetic rats. Methods The animal model of insulin resistance in 2 type diabetic was established by injecting low dose of STZ in 60 high fat-fed SD rats. After one week, the successful models were randomly divided into 4 groups: untreated, high and low OPRA-treated, mefformin-treated group, Meanwhile, a normal control group was designed. After four weeks, the differences of FBG, PG, INS and C-P were compared among the groups. Results The OPRA can significantly reduced the level of serum TC and FBG, increased the C-E ]Lightened the IRI(P<0.01). Conclusion The OPRA can reduce the level of serum TG and FBG, and decrease insulin resistance to protect the β-cell function.     

冬连胶囊对实验性糖尿病大鼠早期胰岛素抵抗的影响

Objective To investigate the effects of Donglian Capsule(the combination of Ophiopogonis Tuber Polysaccharide with Rhizoma Coptidis Alkaloids, OPRA for short, Component of Chinese medicine)on early insulin resistance in experimental diabetic rats. Methods The animal model of insulin resistance in 2 type diabetic was established by injecting low dose of STZ in 60 high fat-fed SD rats. After one week, the successful models were randomly divided into 4 groups: untreated, high and low OPRA-treated, mefformin-treated group, Meanwhile, a normal control group was designed. After four weeks, the differences of FBG, PG, INS and C-P were compared among the groups. Results The OPRA can significantly reduced the level of serum TC and FBG, increased the C-E ]Lightened the IRI(P<0.01). Conclusion The OPRA can reduce the level of serum TG and FBG, and decrease insulin resistance to protect the β-cell function.     

冬连胶囊对实验性糖尿病大鼠早期胰岛素抵抗的影响

Objective To investigate the effects of Donglian Capsule(the combination of Ophiopogonis Tuber Polysaccharide with Rhizoma Coptidis Alkaloids, OPRA for short, Component of Chinese medicine)on early insulin resistance in experimental diabetic rats. Methods The animal model of insulin resistance in 2 type diabetic was established by injecting low dose of STZ in 60 high fat-fed SD rats. After one week, the successful models were randomly divided into 4 groups: untreated, high and low OPRA-treated, mefformin-treated group, Meanwhile, a normal control group was designed. After four weeks, the differences of FBG, PG, INS and C-P were compared among the groups. Results The OPRA can significantly reduced the level of serum TC and FBG, increased the C-E ]Lightened the IRI(P<0.01). Conclusion The OPRA can reduce the level of serum TG and FBG, and decrease insulin resistance to protect the β-cell function.