保肾通络方含药血清调节线粒体自噬对高糖诱导下足细胞氧化损伤的影响

目的 观察保肾通络方含药血清对高糖培养下足细胞氧化损伤的影响,并对保肾通络方的作用机制进行探讨。方法 以体外培养的肾小球足细胞为研究对象,给予高浓度葡萄糖刺激,采用CCK-8法分析足细胞活力以确定最佳模型及给药浓度,分为正常对照组、高糖组、保肾通络方组,采用鬼笔环肽进行细胞骨架染色,Western blotting法检测足细胞裂孔膜蛋白nephrin,自噬相关蛋白5(autophagy-related protein 5,ATG5)、ATG7、微管相关蛋白1轻链3(microtubule-associated protein 1 light chain 3,LC3)的表达,流式细胞术检测足细胞活性氧(reactive oxygen species,ROS)及线粒体膜电位,免疫荧光共标LC3B及线粒体,观察各组足细胞内线粒体自噬泡的生成情况。结果 30 mmol/L高糖干预48 h可显著降低足细胞活力(P＜0.05),保肾通络方含药血清可使高糖诱导的足细胞活力明显提升(P＜0.01);与正常对照组相比,高糖组足细胞骨架褶皱呈多边形,应力纤维束被破坏,ROS生成显著增多(P＜0.01),足细胞nephrin、ATG5、ATG7表达显著降低,LC3Ⅱ/LC3Ⅰ比值降低(P＜0.05),LC3B与线粒体共标减少,线粒体膜电位下降(P＜0.01);与高糖组相比,保肾通络方组足细胞骨架改善,ROS生成减少(P＜0.01),nephrin、ATG5、ATG7表达显著增加(P＜0.05),LC3Ⅱ/LC3Ⅰ比值上升(P＜0.05),LC3B与线粒体荧光共标增多,线粒体膜电位升高(P＜0.01)。结论 保肾通络方含药血清能够减轻高糖诱导的足细胞氧化损伤,其作用可能与改善足细胞线粒体自噬有关。     

六种人工晶状体计算公式预测三焦点人工晶状体屈光度准确性的比较

目的：评估Barrett Universal II计算公式预测三焦点人工晶状体（IOL,AT Lisa tri.839MP）屈光度的准确性,并与其他5种IOL计算公式（Haigis、HofferQ、SRK/T、SRKII和Holladay1）进行对比。方法：前瞻性临床研究。选择2016年12月至2018年8月于武汉爱尔眼科医院行飞秒激光辅助超声乳化白内障吸除联合三焦点IOL植入术的患者。术前使用Lenstar LS 900进行光学生物测量,并通过该仪器内置的其中6种IOL计算公式预测术后屈光度,IOL优化常数取自ULIB网站。随访至术后3个月行主觉验光,比较各IOL计算公式平均绝对屈光误差（MAE）的差异,并对各公式绝对屈光误差中值 （MedAE）、绝对屈光误差最大值（MaxAE）以及术后屈光误差在±0.5 D、±1.0 D和±2.0 D范围内患眼的百分比进行评估比较。数据采用Kruskal-Wallis H检验进行分析。结果：本研究最终纳入患者 55例（78眼）。根据各公式MAE值由小到大排序,依次为Barrett universal II（0.332 D）、SRK/T（0.420 D）、 Haigis（0.480 D）、Holladay1（0.607 D）,HofferQ（0.626 D）以及SRKII（0.822 D）。各公式MAE差异具有统计学意义（P<0.001）。Barrett Universal II公式中MaxAE小于其他IOL计算公式。术后屈光误差在 ±0.5 D、±1.0 D以及±2.0 D范围内患眼所占百分比,Barrett Universal II公式最高,分别为73%、94.8% 以及100%。结论：Barrett Universal II、SRK/T、Haigis公式能更准确地预测患者术后实际屈光结果。因此,对于植入三焦点IOL的患者我们推荐使用Barrett Universal II、SRK/T或者Haigis公式。     

鲜地黄配方颗粒成型工艺优选

目的:探讨鲜地黄配方颗粒的成型工艺.方法:以制粒粒度、堆密度、流动性、溶化性、吸湿性为指标,考察不同辅料种类及辅料用量;以软材性状、制粒难易程度、成品性状为指标,考察不同体积分数乙醇,优选适宜润湿剂.结果:最佳制粒工艺为干燥粉与麦芽糊精按1:1比例混匀,以85％乙醇为润湿剂制软材,1号药筛挤压制粒,60℃干燥,整粒.成品颗粒的临界相对湿度60％.结论:该优选工艺方法简便易行,成品率高,为鲜地黄配方颗粒大生产提供实验依据.     

自制益智方渗透泵片的体外释放度考察

目的:考察激光打孔的孔径与释放条件对自制益智方渗透泵片体外释放的影响。方法:处方量主药与辅料混合均匀,70%乙醇制粒、压片、包衣,衣膜增重7%,熟化,激光打孔器双面打孔,制备孔径为0.2、0.4、0.6、0.8、1.0mm激光打孔片剂。考察不同测定方法(转篮法、桨法)、不同孔径、不同转速、不同渗透压条件下释放度,并比较4种有效成分(人参皂苷Rg1、人参皂苷Rb1、人参皂苷R1、桅子苷)释放度,探索其释放规律。结果:转篮法测定该渗透泵片,释放度重复性较好;激光打孔孔径为0.2-0.6mm时,近恒速释放;在50r/min转速条件下,0.4mm孔径片中4种成分12h累积释放度均达到了85%以上;释药度随着释放介质渗透压的升高而减少;4种检测成分间释放度相似因子均大于50;0.4mm孔径,转篮法50r/min条件下,10h释放较符合零级释放方程y=9.1034t+4.5482(r=0.9915)。结论:转篮法适于测定益智渗透泵片的释放度,激光打孔孔径对释放度有一定影响,衣膜内外渗透压是片剂释放动力,4种有效成分释放度相似,该制剂的释放符合零级释放方程。     

芍药甘草汤及其拆方对H2O2诱导损伤小肠上皮细胞的保护作用

目的:研究芍药甘草汤及其拆方对H2O2诱导损伤大鼠小肠上皮细胞的保护作用。方法:实验设空白对照组,H2O2模型组,白芍(Paeoniae Radix,PR)组,甘草(Licorice,L)组,白芍甘草不同比例组。药物组在药物作用24 h后,与模型组一起加入200μmol.L-1的H2O2作用2 h,空白对照组正常培养,用MTT方法检测细胞活性,Annexin V/PI双染检测细胞凋亡。RT-qPCR检测Caspase-3和Bax mRNA表达。结果:与空白对照组比较,在H2O2损伤后,模型组细胞MTT检测吸光度(A)明显降低,存活细胞明显减少,凋亡增高,Caspase-3表达明显增高,Bax mRNA无明显变化;与H2O2模型组比较,PR 200μg.mL-1,PR 100μg.mL-1,L 100μg.mL-1,L50μg.mL-1,PR 200μg.mL-1/L100μg.mL-1,PR 100μg.mL-1,100μg.mL-1,PR100μg.mL-1,50 mg.L-1组A明显增高,细胞凋亡明显减轻,Caspase-3表达明显降低。结论:白芍,甘草单用,以及白芍与甘草配伍均对H2O2诱导损伤的小肠上皮细胞有保护作用,其机制可能与抑制Caspase-3 mRNA的表达相关。     

Synergistic interaction between Astragali Radix and Rehmanniae Radix in a Chinese herbal formula to promote diabetic wound healing

Ethnopharmacological relevance

Astragali Radix (AR) and Rehmanniae Radix (RR) are two traditional Chinese medicines widely used in China for treating diabetes mellitus and its complications, such as diabetic foot ulcer.

Aim of study

In our previous study, a herbal formula NF3 comprising AR and RR in the ratio of 2:1 was found effective in enhancing diabetic wound healing in rats through the actions of tissue regeneration, angiogenesis promotion and inflammation inhibition. The aims of the present study were to investigate the herb–herb interaction (or the possible synergistic effect) between AR and RR in NF3 to promote diabetic wound healing and to identify the principal herb in the formula by evaluating the potencies of individual AR and RR in different mechanistic studies.

Materials and methods

A chemically induced diabetic foot ulcer rat model was used to examine the wound healing effect of NF3 and its individual herbs AR and RR. For mechanistic studies, murine macrophage cell (RAW 264.7) inflammation, human fibroblast (Hs27) proliferation and human endothelial cell (HMEC-1) migration assays were adopted to investigate the anti-inflammatory, granulation formation and angiogenesis-promoting activities of the herbal extracts, respectively.

Results

In the foot ulcer animal model, neither AR nor RR at clinical relevant dose (0.98 g/kg) promoted diabetic wound healing. However, when they were used in combination as NF3, synergistic interaction was demonstrated, of which NF3 could significantly reduce the wound area of rats when compared to water group (p < 0.01). For anti-inflammation and granulation formation, AR was more effective than RR in inhibiting lipopolysaccharide (LPS)-induced nitric oxide production from RAW 264.7 cells and promoting Hs27 fibroblast proliferation. In the aspect of angiogenesis promotion, only NF3 promoted cell migration of HMEC-1 cells.

Conclusions

AR plays a preeminent role in the anti-inflammatory and fibroblast-proliferating activities of NF3. The inclusion of RR, however, is crucial for NF3 to exert its overall wound-healing as well as the underlying angiogenesis-promoting effects. The results of present study justified the combined usage of AR and RR in the ratio of 2:1 as NF3 to treat diabetic foot ulcer and illustrated that AR is the principal herb in this herbal formula.     

中药配方颗粒的发展现状及国际化对策探讨

近年来,世界各国对天然药物日益重视,而中药配方颗粒既能在中医理论指导下随证加减,又有服用便捷、适宜工业生产等优点,成为推动中药"走出去"的有力助力。通过回顾中药配方颗粒的发展历程、分析相关数据和国内外发展形势,对中药配方颗粒的研究现状、市场情况及国际化发展面临的主要挑战进行探讨,提出利用政府间交往推进中药国际贸易、充分发挥海外资源平台作用、帮助监管部门建立符合中药配方颗粒特点和审批国政策的合理化国际审批方案、争取国际标准主导权的建议,为推动中药配方颗粒产业在世界范围内高质量发展提供参考。     

LASIK术后角膜前表面曲率的分析与预测

目的:比较Orbscan眼前节系统与经验公式所得准分子激光原位角膜磨镶术术后角膜前表面曲率的差异,评价经验公式预测的准确性.方法:对53例106眼在术前和术后1a进行Orbscan角膜曲率测量,用经验公式计算出术后角膜前表面曲率.结果:Orbscan和经验公式所得术后角膜前表面曲率分别为37.63±.38D,36.45±3.68D,差异有统计学意义(P＜0.001).在低度和中度近视组,两种方法所得术后角膜前表面曲率分别为39.18±2.57D,39.92±1.44D,差异无统计学意义(P=0.157);在高度近视组,两种方法所得术后角膜前表面曲率分别为37.10±2.07D,35.37±3.48D,差异有统计学意义(P＜0.001).结论:经验公式能准确预测低、中度近视患者LASIK术后的角膜前表面曲率.     

Determination of furfural and hydroxymethyl furfural from baby formula using dispersive liquid–liquid microextraction coupled with high performance liquid chromatography and method optimization by response surface methodology

A simple, rapid and efficient method was developed using dispersive liquid–liquid microextraction (DLLME) coupled with high performance liquid chromatography (HPLC) for extraction and determination of furfural (F) and hydroxymethyl furfural (HMF) in baby formula. The effects of different variables on the extraction efficiency such as the volume of extraction and disperser solvents, pH and salt effect were studied and optimized simultaneously using (RSM) based on central composite design (CCD). Under optimum conditions, a mixture of ethanol (disperser solvent) and 1-octanol (extraction solvent) was rapidly injected in to the sample solution (after adding 2 g salt and adjusting pH to 6.5). Limit of detection for F and HMF were 0.7 and 1.8 ng g⁻¹, respectively. The inter-day relative standard deviation (RSD%) were 4.9 for HMF and 3.9 for F and also inter-day RSD% were in the range of 5.2–8%. The results showed that DLLME-HPLC is a very fast, simple, sensitive and accurate analytical method for the determination of F and HMF in baby formulas. Finally, The ability of the proposed method to determine F and HMF in different baby formulas in Iran was studied and suitable result was obtained.     

截断逆挽方对慢加急性肝衰竭大鼠肝细胞进入S期关键蛋白的影响

目的:观察截断逆挽方对慢加急性肝衰竭(ACLF)大鼠肝脏细胞进入S期关键蛋白的影响。方法:将150只Wistar大鼠随机分成3组,正常组(NC)、模型组(MC)及截断逆挽方组(JD)。采用人血清白蛋白联合D-氨基半乳糖和脂多糖急性攻击制作ACLF模型。JD组大鼠在急性攻击24 h后连续灌胃,在第5、10、15天平行取材。运用HE染色法观察大鼠肝脏组织结构,Western blot法检测转录因子E2F1的表达量,RT-PCR检测CyclinA、E和Cdc25 mRNA表达量。结果:与模型组相比,JD组大鼠E2F1的表达量明显增高,并在第10天时差异最为显著(P<0.05),CyclinA、E、Cdc25表达量均有不同程度升高;与正常组相比,JD组大鼠10 d、15 d时CyclinA mRNA表达量差异显著,JD组大鼠在10 d、15 d时Cdc25和CyclinE mRNA表达量均差异显著(P<0.05)。结论:截断逆挽方可以通过调节E2F1介导的细胞再生信号通路上的关键蛋白E2F1、CyclinA、CyclinE、Cdc25的表达,从而促使细胞进入S期,加快DNA复制进程,提高肝细胞的增殖率,发挥治疗ACLF的作用。