载As2O3 pH敏感钙砷复合物脂质体的制备及体外评价

目的 制备pH敏感释药的As₂O₃脂质体，并进行体外评价。方法 采用薄膜分散法制备含钙离子脂质体，然后用离子沉淀法孵育制备钙砷复合物脂质体（CaAs-LP）。测定CaAs-LP的粒径、Zeta电位及多分散系数（PDI）；透射电子显微镜观察脂质体的形态；电感耦合等离子体发射光谱仪测定纳米药物的载药量与包封率；透析袋法考察其体外释药特性。噻唑蓝（MTT）法考察未载药脂质体及CaAs-LP对人源性乳腺癌MCF-7细胞、人源性脑胶质瘤U87细胞和人源性肝癌HepG2细胞的毒性；共聚焦显微镜考察U87细胞对CaAs-LP的摄取效率。结果 制备的CaAs-LP呈规整类球型，粒径约为（117.16±1.94）nm，包封率和载药量分别为（74.31±2.11）%、（8.31±0.13）%。体外释放研究表明，CaAs-LP具有明显的缓释以及pH响应释药特征。未载药的脂质体在MCF-7、U87、HepG2和L02细胞中的生物相容性良好；CaAs-LP抑制肿瘤细胞生长的作用较原药有所上升，半数抑制浓度（IC₅₀）值分别为11.91、4.90、19.41、27.59 μmol/L。细胞摄取研究表明肝癌细胞对脂质体具有良好的摄取。结论 CaAs-LP具备显著的缓释以及pH响应释药的特性，在肿瘤治疗方面具有较好的应用前景。     

基于UPLC-LTQ-Orbitrap高分辨质谱的葛根芩连汤的化学成分分析

目的基于超高效液相色谱-离子阱-静电场轨道阱质谱(UPLC-LTQ-Orbitrap-MS)的分析方法鉴定葛根芩连汤中化学成分。方法采用色谱柱Dikma Endeavorsil C18(100 mm×2.1 mm,1.8μm),流动相为0.1%甲酸水(A)-乙腈(B),体积流量0.3 mL/min,梯度洗脱。质谱采用ESI源,正负离子模式分别采集一级、二级质谱数据。结果通过对照品指认、软件预测分析,结合文献报道,从葛根芩连汤中鉴定出67个化合物,包括黄酮类36个、生物碱类12个、三萜类及三萜皂苷类4个及其他15个。结论利用UPLC-LTQ-Orbitrap-MS系统阐明葛根芩连汤中化学成分,并初步归纳其各类主要化学成分的质谱裂解特点,为葛根芩根芩连汤的质量控制和作用机制研究提供了参考依据。     

羌活汤离子导入联合关节松动术治疗膝骨关节炎寒湿痹阻证临床研究

目的评价羌活汤离子导入联合关节松动术治疗膝骨关节炎(knee osteoarthritis,KOA)寒湿痹阻证的疗效。方法将本院2017年10月-2018年10月符合入选标准的98例KOA患者,按随机数字表法分为2组,每组49例。2组均给予关节松动术治疗,在此基础上,对照组口服硫酸氨基葡萄糖胶囊,观察组经离子导入羌活汤。2组均治疗6周。分别于治疗前后从关节疼痛、关节得温痛减、晨僵、腰身重痛方面行寒湿痹阻证症状评分,采用西安大略和麦克马斯特大学骨关节炎指数(The Osteoarthritis Index at the Universities of Western Ontario and McMaster,WOMAC)评估膝关节功能障碍程度,采用ELISA法检测血清Ⅰ型胶原C端肽(typeⅠcollagen C-terminal peptide,CTX-Ⅰ)、CTX-Ⅱ、抗酒石酸酸性磷酸酶5b(tartrate-resistant acid phosphatase isoform 5b,TRACP-5b)及前列腺素E2(prostaglandin E2,PGE2)、P物质(substance P,SP)、多巴胺(dopamine,DA)、5-羟色胺(5-hydroxytryptamine,5-HT)水平,评价临床疗效。结果观察组总有效率为93.9%(46/49)、对照组为77.6%(38/49),2组比较差异有统计学意义(χ^2=4.083,P=0.043)。治疗后,观察组WOMAC指数中关节疼痛、关节僵硬、日常活动受限评分低于对照组(t值分别8.430、9.986、12.776,P<0.01);症状评分中关节疼痛、关节得温痛减、晨僵、腰身重痛评分低于对照组(t值分别为8.825、9.043、7.230、7.034,P<0.01)。治疗后,观察组血清CTX-Ⅰ[(404.99±43.35)μg/L比(458.69±48.61)μg/L,t=21.005]、CTX-Ⅱ[(414.99±43.39)μg/L比(484.06±50.77)μg/L,t=18.991]、TRACP-5b[(2.98±0.35)U/L比(5.67±0.61)U/L,t=9.043]水平均低于对照组(P<0.01);血清PGE2[(167.81±18.79)μg/L比(252.61±27.34)μg/L,t=26.389]、SP[(143.67±15.92)μg/L比(179.55±19.53)μg/L,t=25.416]、DA[(9.15±1.15)μg/L比(13.17±1.81)μg/L,t=10.445]、5-HT[(615.08±63.95)μg/L比(712.69±72.88)μg/L,t=31.004]水平均低于对照组(P<0.01)。结论羌活汤离子导入联合关节松动术可改善KOA患者骨代谢水平,降低疼痛介质分泌,改善临床症状,提高疗效。     

Characterization of Hybrid Materials Prepared by Sol-Gel Method for Biomedical Implementations. A Critical Review

The interaction between tissues and biomaterials (BM) has the purpose of improving and replacing anatomical parts of the human body, avoiding the occurrence of adverse reactions in the host organism. Unfortunately, the early failure of implants cannot be currently avoided, since neither a good mixture of mechanical and chemical characteristics of materials nor their biocompatibility has been yet achieved. Bioactive glasses are recognized to be a fine class of bioactive substances for good repair and replacement. BM interact with living bones through the formation of a hydroxyapatite surface layer that is analogous to bones. Bioglasses’ composition noticeably affects their biological properties, as does the synthesis method, with the best one being the versatile sol-gel technique, which includes the change of scheme from a ‘sol’ fluid into a ‘gel’. This process is widely used to prepare many materials for biomedical implants (e.g., hip and knee prostheses, heart valves, and ceramic, glassy and hybrid materials to serve as carriers for drug release). Nanoparticles prepared by the sol-gel method are interesting systems for biomedical implementations, and particularly useful for cancer therapy. This review provides many examples concerning the synthesis and characterization of the above-mentioned materials either taken from literature and from recently prepared zirconia/polyethylene glycol (PEG) hybrids, and the corresponding results are extensively discussed.     

The effects of metal ion PCR inhibitors on results obtained with the Quantifiler® Human DNA Quantification Kit

Forensic DNA samples may include the presence of PCR inhibitors, even after extraction and purification. Studies have demonstrated that metal ions, co-purified at specific concentrations, inhibit DNA amplifications. Metal ions are endogenous to sample types, such as bone, and can be introduced from environmental sources. In order to examine the effect of metal ions as PCR inhibitors during quantitative real-time PCR, 2800 M DNA was treated with 0.0025–18.750 mM concentrations of aluminum, calcium, copper, iron, nickel, and lead. DNA samples, both untreated and metal-treated, were quantified using the Quantifiler^® Human DNA Quantification Kit. Quantification cycle (Cq) values for the Quantifiler^® Human DNA and internal PCR control (IPC) assays were measured and the estimated concentrations of human DNA were obtained. Comparisons were conducted between metal-treated and control DNA samples to determine the accuracy of the quantification estimates and to test the efficacy of the IPC inhibition detection. This kit is most resistant to the presence of calcium as compared to all metals tested; the maximum concentration tested does not affect the amplification of the IPC or quantification of the sample. This kit is most sensitive to the presence of aluminum; concentrations greater than 0.0750 mM negatively affected the quantification, although the IPC assay accurately assessed the presence of PCR inhibition. The Quantifiler^® Human DNA Quantification Kit accurately quantifies human DNA in the presence of 0.5000 mM copper, iron, nickel, and lead; however, the IPC does not indicate the presence of PCR inhibition at this concentration of these metals. Unexpectedly, estimates of DNA quantity in samples treated with 18.750 mM copper yielded values in excess of the actual concentration of DNA in the samples; fluorescence spectroscopy experiments indicated this increase was not a direct interaction between the copper metal and 6-FAM dye used to label the probe that targets human DNA in the Quantifiler^® kit. Evidence of inhibition was observed for the human-specific assay at a lower metal concentration than detected by the IPC, for all metals examined except calcium. These results strongly suggest that determination of a “true negative” sample should not be based solely on the failure of the IPC to indicate the presence of a PCR inhibitor and indicate that amplification of all samples should be attempted, regardless of the quantification results.     

Bone resorption and serum levels of vitamin D metabolites in the hypophosphataemic rat

The supplementation of a low phosphate diet with vitamin D has been shown to result in an increase in bone resorption in the hypophosphataemic rat. The aim of the present study was to determine if administration of vitamin D to rats fed a vitamin D and phosphate-depleted diet would result in an increase in the circulatory levels of the active vitamin D metabolite 1,25(OH)₂D₃ and an associated increase in bone resorption. Three groups of weanling Sprague-Dawley rats were used. The first group consisted of control animals on a normal laboratory stock diet and the second and third groups were experimental animals receiving a vitamin D-and phosphate-deficient diet with the third group receiving vitamin D supplementation. All animals were housed in the dark. After 30 days on the diet the experimental animals received 0.1 mmol NaH₂PO₄ by intraperitoneal injection. Blood was sampled at zero, 3, 6, 18 and 48 h post-injection and analysed for the vitamin D metabolites 25(OH)D₃ and 1,25(OH)₂D₃, calcium and inorganic phosphate (Pi). The serum analyses revealed that the level of 25(OH)D₃ in the hypophosphataemic animals was significantly lower than that of the control animals. However the 1,25(OH)₃D₃ level was initially significantly higher, then dropped to the control level at 18 h post-intraperitoneal injection of phosphate. Further, the serum levels of 25(OH)D₃ and 1,25(OH)₂D₃, calcium and Pi in the hypophosphataemic animals supplemented with vitamin D were significantly higher than those of the vitamin D-deficient animals. Also the vitamin D-supplemented animals exhibited significantly greater levels of bone resorption. These results therefore, are consistent with a role of 1,25(OH)₂D₃ in bone resorption in hypophosphataemic rats.     

无机抗菌剂对自酸蚀处理剂全身毒性和细胞毒性影响的研究

目的测试无机抗菌剂的添加对新型实验自酸蚀处理剂（ESP）全身毒性和细胞毒性的影响。方法将6种无机抗菌剂分别加入ESP中，进行经口途径大鼠短期全身毒性实验。并采用受试材料与NIH3T3细胞直接接触培养的方法，观测其对细胞增殖、活力和形态的影响。结果抗菌剂添加比为5.0%时，为期2周的大鼠全身毒性实验中未发现临床毒性体征，各组间大鼠体重相对增长率无显著差异。经含质量浓度为0.5%不同抗菌剂的处理剂处理固化后的各实验组细胞活力比率和细胞增殖率与ESP空白对照组相比无显著差异。仅样本覆盖区及样本边缘邻近区域细胞可观察到毒性表现，远离样本覆盖区的细胞密度未受明显影响。结论以适当的质量浓度添加受试的无机抗菌剂对受试ESP的全身毒性和细胞毒性无显著影响。