牛磺鹅去氧胆酸钠的提取及与鹅去氧胆酸钠的药理作用比较

以500 mL鸡胆汁为基本原料,采用沉淀、过滤、减压蒸馏、真空干燥、抽提、重结晶法,制得了7.2 g结合型的牛磺鹅去氧胆酸钠(TCDCANa),以TLC及IR法确定其结构。并将其与鹅去氧胆酸钠(CDCANa)进行镇咳、平喘和祛痰作用对比实验。药理实验结果为:TCDCANa和CDCANa的镇咳作用减咳率分别为(29.1±5.6)%和(18.2±6.8)%;平喘作用解痉率分别为(79.9±22.8)%和(73.3±22.7)%,祛痰作用酚红排泌量分别为(1.601±0.391)μg/mL和(1.029±0.308)μg/mL,TCDCANa与CDCANa间存在显著性差异(P<0.01)。药理实验结果表明,TCDCANa具有明显的镇咳、平喘、祛痰和抗炎作用,其作用强于CDCANa。     

牛磺胆酸钠的制备及与胆酸的药效对比

以胆酸和牛磺酸为基本原料制备了牛磺胆酸钠 (TCANa) ,收率超过 5 9% ,w(TCANa) >92 % ,其IR和1HNMR与结构相符。并将牛磺胆酸钠与胆酸钠进行镇咳、祛痰和平喘对比实验。药理实验结果为 :牛磺胆酸钠和胆酸钠的镇咳作用减咳率分别为 (2 8 6± 6 7) %和 (18 2±6 8) % ;祛痰作用酚红排泌量分别为 (1 4 82± 0 4 0 1) μg/mL和 (0 95 9± 0 312 ) μg/mL ;平喘作用解痉率分别为 (71 2± 2 3 1) %和 (71 2± 2 3 1) % ,与生理盐水间存在着显著性差异 ,牛磺胆酸钠与胆酸钠间也存在显著性差异。药理实验表明牛磺胆酸钠具明显的镇咳、祛痰和平喘作用 ,其作用强于胆酸     

ELISA法间接检测血管内皮生长因子抑制剂的相对结合力

目的采用ELISA法间接检测血管内皮生长因子抑制剂(VEGF Trap)的相对结合力。方法以固定量的VEGF165与VEGF Trap结合,通过双抗体夹心ELISA法检测未结合的、游离的VEGF165,进而求得VEGFTrap的半数抑制浓度(IC50),计算其与反应体系中参比品的IC50之比,即为VEGFTrap的相对结合力。结果同一批次的3支VEGF Trap样品分别经3次测定,相对结合力分别为(100.86±9.43)%、(88.46±13.94)%和(82.66±9.60)%,均值为(90.66±12.60)%,符合该制品质量标准的要求。3次试验的变异系数分别为9.35%、15.76%和11.61%。结论 ELISA法精密性良好,结果客观,影响因素少,可用于VEGF Trap相对亲和力的常规检测。     

鸟嘌呤和腺嘌呤吸附和电氧化行为的电化学石英晶体微天平研究

基于压电电化学石英晶体微天平(EQCM)技术,研究了腺嘌呤(A)和鸟嘌呤(G)在裸金电极和多壁碳纳米管(MWCNTs)修饰金电极上的吸附和电氧化行为。结果表明,在金电极上,A的吸附量和氧化电流均比G大,而在MWCNTs/Au电极上,A和G可类似地吸附,但其氧化峰电位负移且氧化峰电流增大,表明MWCNTs对A和G的氧化具有催化作用。实验发现,A和G在裸金电极上氧化的电子转移数分别为5.4(RSD=±2.3%)和1.9(RSD=±1.3%),而在MWCNTs/Au电极上的氧化电子转移数分别为5.1(RSD=±1.8%)和1.5(RSD=±1.6%)。     

靶向高迁移率族蛋白A1基因RNA干扰真核表达载体的构建

目的构建针对人高迁移率族蛋白A1(HMGA1)基因的RNA干扰真核表达载体,为研究HMGA1基因在肿瘤细胞中的作用奠定实验基础。方法设计合成特异性针对人HMGA1基因的寡核苷酸序列,梯度退火后,与pU6mRFP载体连接,构建重组载体,转染人肝癌细胞SMMC-7721。通过激光共聚焦显微镜观察红色荧光,估测转染效率,RT-PCR法检测转染细胞HMGA1mRNA水平,流式细胞仪检测细胞凋亡和细胞周期。结果DNA测序证实,成功构建了特异性HMGA1siRNA真核表达载体,转染后72h,转染效率为40%左右。所构建的载体能够特异性沉默转染细胞HMGA1基因的表达。转染细胞HMGA1基因沉默后,细胞凋亡率(27·86%±2·44%)明显高于空载体对照组和SMMC-7721对照组(分别为2·82%±2·39%和2·04%±0·70%),G0-G1期细胞百分数(77·73%±1·78%)明显高于空载体对照组和SMMC-7721对照组(分别为42·19%±3·28%和39·23%±3·63%),G2-S期细胞百分数(22·27%±1·78%)明显低于空载体对照组和SMMC-7721对照组(分别为57·81%±3·28%和60·77%±3·63%)。结论成功构建了HMGA1的RNA干扰真核表达载体。     

AoGDW肽抑制血小板聚集的活性及特异性

目的研究精氨酸-甘氨酸-天门冬氨酸(RGD)肽衍生物——ω-氨基辛酸-甘氨酸-天门冬氨酸-色氨酸(AoG-DW)抗血小板聚集的活性及特异性。方法采用比浊法测定AoGDW肽抑制人血小板聚集的活性,体外细胞脱黏附试验及MTT法检测AoGDW对人脐带静脉内皮细胞(HUVEC)的脱黏附作用。结果AoGDW和RGDS抑制人血小板聚集的IC50分别为(4.71±2.51)μmol/L和(61.82±8.08)μmol/L,AoGDW使HUVEC产生脱黏附的IC50为(16.50±0.68)mmol/L。结论AoGDW具有高活性的抗血小板聚集作用,并保留了其特异性,有可能避免或减少毒副作用的发生。对于抗血小板药物的临床应用将具有重要的现实意义。     

YKL-40转染对前列腺癌LNcap细胞增殖及侵袭活性的影响

目的探讨外源性YKL-40基因转染对前列腺癌LNcap细胞增殖、侵袭、迁移及黏附活性的影响。方法 RT-PCR法检测前列腺癌细胞LNcap、PC-3、DU-145 YKL-40基因内源性表达情况;用pcDNA3.1-YKL-40质粒转染LNcap细胞,分别经MTT法、Boyden小室法及黏附试验检测转染前后细胞增殖、侵袭、迁移及黏附活性,并进行体外药敏试验。结果仅DU-145细胞可内源性表达YKL-40基因;pcDNA3.1-YKL-40转染的LNcap细胞在第2～5天的A490值均显著高于对照组(P<0.05),其侵袭(75.11±4.40)和迁移穿膜细胞数(133.00±5.07)及黏附率(107.57%)均显著高于对照组(P<0.05),对5-FU、顺铂和依托泊苷的IC50值分别为(31.15±0.43)、(4.15±0.13)和(55.22±0.57)μmol/L,均显著高于对照组(P<0.05)。结论 YKL-40基因能促进LNcap细胞增殖,提高细胞侵袭、迁移及黏附活性,并使其对5-FU、顺铂和依托泊苷具有一定的耐药性。     

狂犬病病毒核蛋白的原核表达及其免疫原性

目的原核表达、纯化狂犬病病毒(Rabies virus,RV)核蛋白(Nucleoprotein,NP),并检测其免疫原性。方法 RT-PCR扩增RV CVS-11株NP全长基因序列,克隆至原核表达载体pET-30a(+),构建重组原核表达质粒pET-30a-N,转化E.coliBL21(DE3),IPTG诱导表达,表达的重组蛋白经Ni2+柱亲和层析纯化后,进行Western blot分析,并分别以A(lOH)3和CpG1826作为佐剂经腹腔免疫BALB/c小鼠,ELISA法检测小鼠的血清特异性抗体水平。结果重组原核表达质粒pET-30a-N经双酶切和测序证实构建正确;表达的重组蛋白相对分子质量约51 800,表达量约占菌体总蛋白的35%,主要以包涵体形式存在;纯化的重组蛋白纯度达90%以上,可与抗RV小鼠血清特异性结合,分别以A(lOH)3和CpG1826作为佐剂免疫小鼠的血清特异性抗体滴度对数的GMT值(3.81±0.22和3.68±0.20)明显高于阴性对照组(1.25±0.22),且差异均有统计学意义(P<0.01)。结论已成功表达并纯化了RV NP,其免疫原性良好,为进一步研究其功能奠定了基础。     

异黄酮衍生物的合成及其活性研究

以间苯二酚和对甲氧基苯乙酸为原料,在三氟化硼乙醚、N,N-二甲基甲酰胺和甲基磺酰氯作用下,采用超声波辐射技术"一锅法"合成了7-羟基-4′-甲氧基异黄酮(Ⅰ),收率为84.0%。然后,将其分别与苯甲酰氯、乙酰氯、苯氧乙酰氯、肉桂酰氯和水杨酰氯成酯得到5个异黄酮衍生物(Ⅱa～e),收率分别为57.1%、60.6%、59.4%、61.1%和50.4%。利用IR、1HNMR、13CNMR和MS对Ⅱa～e的结构进行表征。体外抗骨质疏松活性实验表明,5种化合物对体外培养的乳鼠颅盖骨成骨细胞的生长有较好的促进作用。     

用自动系统凯氏定氮仪测定生物制品中蛋白质含量

应用自动系统凯氏定氮仪测定人血白蛋白和人血丙种球蛋白的蛋白含量 ,重复性测定X % (n =10 )±SD分别为 2 0 45 %± 0 18%和 10 2 5 %± 0 12 % ;用硫酸铵、氨基乙酸和标准蛋白质控液作回收率试验R % (n =10 )±SD分别为 10 0 .2 4%± 0 5 7% ,99.89%± 0 43%和 10 0 5 0 %± 0 75 % ;用该法测定蛋白质含量与手工定氮法比较有良好相关性 ,总氮 (TN)相关系数r(n =2 6 )为 0 9999;非蛋白氮 (NPN)r(n =19)为 0 9946 ;蛋白氮 (PN)r(n =19)为 0 .9999。测定同批标准人血白蛋白 ,结果X % (n =2 0 )±SD为 5 5 3%± 0 14% ,与其参考标准值 5 5 4%± 0 12 %非常接近。因此 ,用该仪器法测定生物制品中的蛋白质含量准确可靠 ,且快速。     

Corrosion inhibition of carbon steel in artificial seawater by caprylate

The corrosion inhibition efficiency of sodium caprylate (SC) in protecting carbon steel (CS) in aqueous 3.5 wt.% NaCl was studied. SC was found to enable good corrosion protection of CS, yielding a maximum corrosion inhibition efficiency of 80 ± 2%. The inhibition efficiency was shown to be directly proportional to the SC surface coverage, characterized by a two-step adsorption isotherm. The adsorption of SC on CS in the lower SC concentration region was found to follow the Langmuir adsorption isotherm and the first-order reversible adsorption kinetics. The corresponding Gibbs free energy of SC adsorption was determined to be −23.8 kJ · mol⁻¹, indicating a highly spontaneous adsorption process. The scanning electron microscopy imaging of the SC-inhibited CS showed that the surface exhibited good corrosion stability during a 72 h exposure to the test medium.     

8种中药提取物在膏霜类化妆品中的抑菌效能研究

采用纤维素酶法提取8种中药(丁香、鱼腥草、虎杖、黄连、大黄、细辛、甘草和黄芩),并采用滤纸片法和试管二倍稀释法研究8种中药提取物的抑菌效能。根据其抑菌作用进行复配研究,参考美国个人护理品委员会(PCPC)的抑菌效能测试方法对膏霜配方进行微生物挑战试验。结果表明,在8种中药提取物中,黄连提取物对金黄色葡萄球菌、铜绿假单胞菌和大肠埃希氏菌的抑制效果最好,其抑菌圈直径分别为(12.4±0.4),(11.4±0.6)和(10.7±0.2)mm,最低抑菌浓度(MIC)分别为31.25,31.25和15.625 g/L。当黄连和黄芩提取物在膏霜配方中的质量分数分别为0.8%和0.6%时,可较强地抑制细菌和真菌的生长,可通过微生物挑战试验。     

Synthesis of quaternary piperazine methacrylate homopolymers and their antibiofilm and anti-quorum sensing effects on pathogenic bacteria

In this work, poly(glycidyl methacrylate) (PGMA, G) homopolymers with different molecular weights (5600–21,000 gmol⁻¹) were synthesized by atom transfer radical polymerization (ATRP) method. Epoxy groups of PGMA homopolymers were converted to hydroxyl and 1-methyl piperazin residues by reacting with 1-methyl piperazine via ring-opening reaction. The obtained poly(2-hydroxy-3-methyl piperazinepropyl methacrylate) (PHMPPMA, G-P) homopolymers (from 9450 to 35,900 gmol⁻¹) were reacted with alkyl halides of different carbon chain lengths (methyl-, ethyl-, propyl-, butyl-, pentyl-iodides, hexyl bromide) and benzyl chloride (arylhalide) to form quaternary PHMPPMA homopolymers (G-QP). MIC values varied from 0.125 mg/ml to 5 mg/ml. It was found that homopolymer (1G-6QP) produced by the modification of PHMPPMA with hexyl iodide with low molecular weight had highest quorum-sensing inhibition zone ranging from 14.5 ± 0.5 mm (MIC) and 10.0 ± 0.5 mm (MIC/4) as well as best violacein inhibition 61.0 ± 2.2% (MIC) to 17.3 ± 0.6% (MIC/8). Homopolymer (1G-BzQP) produced by the modification of PHMPPMA with benzyl chloride with the lowest molecular weight had best biofilm inhibition ranging from 77.83 ± 1.60% (MIC) to 18.23 ± 0.87% (MIC/8) on S. aureus, 75.70 ± 5.20% (MIC) to 14.25 ± 1.10% (MIC/8) on E. coli and from 86.16 ± 0.83% (MIC) to 12.41 ± 0.76% (MIC/8) on C. albicans. The synthesized homopolymers can be used to combat microbial resistance and severity of infections.     

Effects of N‐trimethyl chitosan chloride as an absorption enhancer on properties of insulin liquid suppository in vitro and in vivo

N‐trimethyl chitosan chloride (TMC), with degree of quaternization (DQ) of 42.77% (TMC40) and 63.03% (TMC60), respectively, were synthesized and used as absorption enhancers in an insulin rectal liquid suppository from poloxamers. The effects of the TMCs on the liquid suppository were investigated. Compared with sodium salicylate (10%), which increased the gelation temperature and decreased the gel strength and bioadhensive force of the liquid suppository, both TMCs increased the three indices due to hydrogen bonding between the amino groups of TMCs and hydroxyl groups of poloxamers. The higher the concentration of TMCs was, the higher the three indices were. At the same concentration, the enhancing effect of TMC60 was higher than that of TMC40. Compared with those of 10% sodium salicylate, the enhancing effects of TMC40 with concentrations of 0.05% and 0.10% and TMC60 with concentration of 0.05% were weaker, while the other testing concentrations all showed better absorption enhancing ability with lower plasma glucose levels, AUC_{0–4 h} (the area below basal glucose level) and C_nadir (the plasma glucose levels at the nadir). Histological assessment was performed by observing irritation of the liquid suppository on the rectal tissues and observation rate of three types of gland changes in the rectal epithelium. Compared with the control (75.48 ± 16.76), TMC40 and TMC60 exhibited little change in observation rates of normal gland (72.10 ± 10.24 and 71.93 ± 9.88, respectively), while sodium salicylate showed significant lower observation rates of normal gland (51.28 ± 13.44). The insulin rectal liquid suppository with TMCs is more effective and safer than that with sodium salicylate. © 2005 Wiley Periodicals, Inc. J Appl Polym Sci 99: 1140–1146, 2006     

pH值和盐浓度对金属螯合亲和层析分离人胰高血糖素样肽-1融合蛋白的影响

目的探讨pH值和盐浓度对金属螯合亲和层析(Immobilized metal ion affinity chromatography,IMAC)分离含His-Tag标签的人胰高血糖素样肽-1(Glucagon-like peptide-1,GLP-1)融合蛋白的影响,并确定最佳洗脱条件。方法在0.50mol/L盐浓度条件下,分别取pH6.0、7.0、7.8和9.04种缓冲液进行咪唑梯度洗脱;在合适的pH值条件下,分别取氯化钠浓度0.25、0.50和0.75mol/L3种缓冲液进行咪唑梯度洗脱。分析在不同pH值及盐浓度条件下洗脱融合蛋白所需要的咪唑浓度及回收率。结果在所考察的pH值范围内,洗脱融合蛋白所需的咪唑浓度随洗脱液pH值的升高而逐渐降低,当洗脱液pH值为7.8时,融合蛋白的分离效果最佳,大部分杂蛋白被0～40mmol/L咪唑洗脱液去除且不含融合蛋白,而绝大部分融合蛋白在咪唑浓度达80～200mmol/L时被洗出,总回收率达(83.7±1.0)%,且比例较高;在此pH值条件下,氯化钠浓度达0.75mol/L,才会影响融合蛋白的回收率,氯化钠浓度为0.25或0.50mol/L的洗脱液洗出融合蛋白的效果相近;在pH7.8,含0.25mol/L氯化钠的洗脱体系下,用50mmol/L咪唑溶液洗脱杂蛋白,200mmol/L咪唑溶液洗脱融合蛋白,融合蛋白的回收率和比例分别可达(85.2±2.0)%和(80.5±1.0)%。结论在所考察的pH值和盐浓度范围内,提高洗脱液pH值和盐浓度均可降低洗脱融合蛋白需要的咪唑量,但也会降低融合蛋白的回收率。     

Laboratory and modelling study of the hygroscopic properties of two model humic acid aerosol particles

The hygroscopic properties of aerosol particles, composed of two different commercially available humic acids, have been investigated in the temperature range 293–295 K as a function of relative humidity using an electrodynamic balance. The measured mass growth factors of Sigma-Aldrich humic acid sodium salt (NaHA) and leonardite humic acid (LHA) at 90% relative humidity are 1.87±0.02 and 1.20±0.05, respectively. There is a general agreement between the results described in this study with previous TDMA results, which are undertaken with smaller particle ensembles and shorter aerosol–water equilibration times. The measured mass growth factors of Sigma-Aldrich humic acid sodium salt (NaHA) and leonardite humic acid (LHA) at 90% relative humidity are 1.87±0.05 and 1.20±0.05, respectively. The UNIFAC model was used to calculate the relationship between water activity and the concentrations of a modelled LHA molecule, and thus the hygroscopicity. The LHA molecule was modelled by both the Steelink monomer and the Northeastern–Temple–Birmingham humic acid monomer structural models.     

Intracellular polysaccharide and its antioxidant activity by Cordyceps militaris SU‐08

The extraction conditions of intracellular polysaccharide from Cordyceps militaris SU‐08 mycelia in submerged culture were investigated. Four parameters affecting the IPS extraction were determined by Plackett‐Burman (PB) tests and then optimized by response surface methodology (RSM). The optimum conditions of IPS extraction were predicted to be, ultrasonic treatment number 61.45, ultrasonic power 543.64 W, ethanol multiple 3.28, and extraction temperature 82.61°C, and the extraction rate of IPS was estimated at 9.11%. The actual value of IPS under these conditions was 9.19%. The in vitro antioxidant results showed that the inhibition effects of IPS at a dosage of 5 g L⁻¹ on hydroxyl, superoxide anion, and 1,1‐diphenyl‐2‐picrylhydrazyl (DPPH) radical were 51.89% ± 3.27%, 82.26% ± 5.03%, and 74.59% ± 4.53%, respectively, which were 32.98% ± 2.71%, 69.71% ± 4.24%, and 41.64% ± 3.28% higher than that of control, respectively. The reducing power of IPS was 0.79 ± 0.03 (absorbance at 700 nm), 11.27% ± 0.82% higher than that of control. The results provide a reference for large‐scale extraction of IPS by C. militaris SU‐08 in industrial fermentation and the IPS can be used as a potential antioxidant which enhances adaptive immune responses. © 2010 Wiley Periodicals, Inc. J Appl Polym Sci, 2011     

乙二醇水溶液腐蚀抑制剂的性能研究

研究了由WH-8缓释剂、钼酸钠、消泡剂SA、有机硅氧烷稳定剂等组成的乙二醇水溶液腐蚀抑制剂的缓蚀性能。结果表明,在33.3%乙二醇溶液中,温度为(88±2)℃,14 d,用量为1.0%时,该腐蚀抑制剂对焊锡、铝、黄铜、紫铜、钢、铁都可以起到优异的腐蚀抑制作用,达到乙二醇型冷却液标准SH 0521—1999的要求。