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。     

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。     

Rapid immunological detection of copper ions using fluorescence immunochromatographic assay

One-step fluorescence immunochromatographic assay (FICA) has been developed using fluorescent microspheres-labelled monoclonal antibody (McAb) probe for the rapid detection of copper ions in herbal plants and soil samples were investigated in this study. The results can be read by the fluorescence reader according to the fitted standard curve equation. Under optimal conditions, detecting copper ions with FICA can be done within 15?min with a LOD (limit of detection) of 0.2?μg/L, and was very useful for the rapid onsite testing. A total of 31 solid samples were pre-treated by microwave digestion combined with cation exchange solid phase extraction and prepared as test samples for FICA and ICP-AES. The two methods showed good relativity with correlation coefficients of 0.924 and 0.969 in herbal plants and soil samples for copper detection, respectively. We hope that our research can help promote the daily monitoring of heavy metal pollution effectively.     

Combined effect of zinc ions and cationic antibacterial agents on intraoral volatile sulphur compounds (VSC)

OBJECTIVE: Volatile sulphur compounds (VSC) are major components of oral malodour. As both zinc ions and cationic antibacterial agents inhibit the formation of oral VSC, this study aimed to determine whether these agents combined have synergistic anti-VSC actions. METHODS: Baseline oral VSC measurements of mouth air from 10 volunteers following cysteine rinsing (6mM, pH 7.2) were obtained using gas chromatography (GC). Subjects rinsed for 1 min with 10ml of the test solutions, 0.3% zinc acetate (Zn), 0.025% chlorhexidine (CHX), 0.025% cetyl pyridinium (CPC), and the combinations Zn+CHX and Zn+CPC. Cysteine rinses were repeated at 1h, 2h and 3h and VSC measurements recorded. Three subjects rinsed with the Zn+CHX combination and fasted for 9h, undergoing cysteine rinses and VSC measurements at 3h intervals. 10 microl of the test solutions were also added to 1ml aliquots of human whole saliva (n=8). Following incubation at 37 degrees C for 24h VSC levels in the saliva headspace were measured by GC. Inhibition of VSC formation and the fractional inhibitory index indicating synergy were calculated. RESULTS: Zn+CHX mouthrinse had a synergistic anti-VSC effect, and was effective for at least 9h. Zn+CPC mouthrinse was less effective. Both combinations showed a synergistic inhibiting effect in-vitro. CONCLUSIONS: Synergy between Zn and the antibacterial agents confirms different mechanisms of operation.     

An in vitro study of the pH of three calcium hydroxide dressing materials

Abstract –  Calcium hydroxide is widely used as a root canal dressing material because of its favorable alkalinizing effect. It has been suggested that the action comes from diffusion of hydroxyl ions through the apical foramen. The purpose of this in vitro study was to test the pH changes that occurred over a period of 30 days using a mixture of calcium hydroxide and distilled water and two commercial calcium hydroxide products in a simulated periapical environment. The materials were inserted in glass tubes closed at one end, which were placed in individual vials containing distilled water at a pH 7.4. Unfilled glass tubes were used as controls. Alkalinity changes of the medium were measured at 1 and 24 h and 15 and 30 days. The alkalinizing properties of all materials showed a rapid increase at 1 and 24 h followed by a continuous but more gradual increase from 15 to 30 days. The control tubes did not cause a change in pH of the medium, which remained at pH 7.4. At the end of the observation period, the alkalinizing properties of Calasept and Ultracal XS were significantly higher ( P  < 0.05) than the calcium hydroxide/distilled water paste.     

Antibacterial activity of silver inorganic agent YDA filler

YDA filler is an antibacterial agent that is currently in commercial dental use. In this study, we attempted to determine whether it exerts an antibacterial effect on human saliva bacteria, and to determine whether it can be used in dental materials. CFUs in 1 mL stimulated human saliva were examined using blood agar and mitis salivarius agar after immersion, with or without YDA filler. The antibacterial effect was compared with that of Ketac-Silver. Dental materials containing 5% wt YDA filler were prepared for in vitro testing on S. mutans and A. viscosus. Furthermore, we examined the in vitro cytotoxicity of experimental MMA resin containing YDA filler on HeLa cells. Human saliva bacteria and mutans streptococci showed reduced viability following exposure to YDA filler after 12 h. The concentration of silver ions released by YDA filler was below 1 ppm after 12 h. Two tested strains showed reduced viability following exposure to dental materials containing YDA filler. In another experiment, MMA resin containing YDA filler did not show cytotoxicity on HeLa cells after 24- and 48-h exposure. Thus, YDA filler may help in the development of antibacterial dental materials, such as composite resin, glass-ionomer or temporary cement.     

Alzheimer amyloid β-peptides exhibit ionophore-like properties in human erythrocytes

Abstract. There is growing evidence that the amyloid β-peptide (β₁_₄₀) is involved in the aetiology of Alzheimer's disease also implicating an altered calcium homeostasis of affected cells. Beta₁_₄₀ has been proposed to form calcium channels in synthetic bilayer membranes [1]. We wanted to investigate in the present study whether β₁-₄₀ (or fragments thereof) could act as ionophores in a biological membrane like the one in human erythrocytes. Incubation of the cells for 2h and 4h at 37°C together with 6μmolL^-1 of β₁-₄₀ or of fragments β₁_₂₈and β₂₅-₃₅, resulted in a significantly decreased energy charge qualitatively similar to the one obtained by a known calcium ionophore (A 23187, 0.05μmolL^-1). Moreover, β₁_₄₀ and its two fragments induced a significant alteration of ⁴⁵Ca permeability in human red blood cells of the same type as the one achieved by the calcium ionophore. The ionophoric action of β₁_₄₀ and its two fragments may lead to an increase of the intracellular calcium ion concentration, in turn resulting in enhanced Ca²⁺-ATPase activity and a decrease in energy charge. This may be valid also for neuronal plasma membranes and could, therefore, be a possible aetiological mechanism in Alzheimer's disease.