Establishment of an Evaluation Method for Gene Silencing by Serial Pulmonary Administration of siRNA and pDNA Powders: Naked siRNA Inhalation Powder Suppresses Luciferase Gene Expression in the Lung

In order to evaluate the in vivo effect of inhaled formulations, it is a gold standard to create a lung metastasis model by intravenously injecting cancer cells into an animal. Because the cancer grows from the blood vessel side, there is a possibility of underestimating the effect of an inhaled formulation administered to the lung epithelium side. In addition, the metastasis model has disadvantages in terms of preparation time and expense. The present study aimed to establish a new method to evaluate the effect of an inhaled small interfering RNA (siRNA) formulation that is more correct, more rapid, and less expensive. We investigated whether siRNA can suppress gene expression of plasmid DNA (pDNA) by serial pulmonary administration of siRNA and pDNA powders prepared by spray-freeze-drying. We revealed that formulations of dry siRNA powder significantly suppressed gene expression of pDNA powder compared with a control group with no siRNA. Naked siRNA inhalation powder with no vector showed the suppression of gene expression equivalent to that of an siRNA-polyethyleneimine complex without damaging tissues. These results show that the present method is suitable for evaluating the gene-silencing effect of inhaled siRNA powders.     

粉防己碱和氯丙嗪对人胚肺成纤维细胞胶原和透明质酸合成的影响(英文)

目的：研究粉防己碱(Tet)和氯丙嗪(Chl)对人胚肺成纤维细胞胶原与透明质酸(HA)合成的影响,为应用钙拮抗剂防治器官纤维化提供依据.方法:采用[~3H]脯氨酸掺入和放射免疫法分别测定胶原与HA合成.结果:Tet 5—80 μmol L~(-1)和Chl 10—40 μmol L~(-1)均以浓度依赖方式抑制胶原与HA合成.Tet 5—20μmol L~(-1))对细胞无明显毒性却显著抑制胶原与HA合成(P<0.01).结论:Tet在低浓度(5—20μmol L~(-1))时对细胞无明显毒性作用而显著抑制成纤维细胞胶原与HA合成,可望成为治疗器官纤维化的有效药物.     

Efficiency and durability of hyaluronic acid of different particle sizes as an injectable material for VF augmentation

Conclusion: The results of the present investigation suggest that modification of HA could improve efficiency and durability in augmentation laryngoplasty. Objectives: Injection laryngoplasty (IL) is one of the most suitable options for treatment of glottic insufficiency, which is caused by vocal fold (VF) paralysis, atrophy, or scarring. Hyaluronic acid (HA) is a widely used material for VF injection. This study was intended to evaluate the durability and efficiency of HA of different particle sizes for VF augmentation. Methods: Three types of HA, Restylane®, monophasic low-viscosity, and unequal particle-sized middle-viscosity HA were injected into the left VF of three groups with eight rabbits each. Results: After 6 and 10 weeks, the injected site was evaluated endoscopically, histologically, radiologically, and functionally. None of the 24 rabbits showed any signs of respiratory distress. Computed tomography (CT) images and endoscopic evaluation revealed sufficient augmented volume of the injected VF in all treated groups 6 weeks after the injection. Histological data at week 10 showed that unequal particle-sized HA did not migrate from its original injection site, while other HAs migrated to the periphery of the arytenoid cartilage. Videokymographic analysis showed more favorable vibrations of unequal particle-sized HA injected VF mucosa 10 weeks post-injection, compared to the other treatment groups.     

透明质酸的瘤学分布与p-CLP示踪学研究

目的:确定透明质酸(HA)瘤内分布,为肺肿瘤的早期诊断提供依据。方法:免疫组化定位肿瘤内HA的分布,亲和层析法从仔猪肋软骨提取猪软骨链蛋白(p-CLP),125I标记p-CLP进行体内分布代谢示踪学研究。结果:HA免疫显色提示肿瘤细胞外基质HA有较高表达,放射示踪实验示1～7h内实验组单位质量的放射性计数百分比(13.46%～29.14%)明显高于对照组(6.4%～13.70%),差异有统计学意义,P<0.01。结论:p-CLP在肺肿瘤内浓集,125I标记p-CLP体内代谢示踪能够为肺肿瘤等疾病的早期诊断提供实验依据。     

透明质酸修饰的尿酸酶多囊脂质体的体外特性及药效学过程

目的 研究透明质酸修饰的尿酸酶(UC)多囊脂质体(uricase in hyaluronic acid-uricase multivesicular liposomes,UHMVLs)的体外特性及其在大鼠体内的药效学.方法 采用复乳法制备UHMVLs并测定包封率及理化特性.取12只健康雄性SD大鼠,模型组采用次黄嘌呤和氧嗪酸钾建立高尿酸血症大鼠模型(n=3),UHMVLs组(n=3)和UC组(n=3)分别于建模后尾静脉注射UHMVLs和游离UC,并以正常组(n=3)为对照,测定大鼠血清中尿酸水平.结果 UHMVLs的平均包封率为(62.48±3.87)％ (n=3).UHMVLs和UC最适温度均为40℃,最适pH值分别为8.0和8.5.在同一温度(20～70℃)和pH(6.5～9.5)条件下,UHMVLs中UC的活性均高于游离UC(P＜0.05).除1、36、48 h外,其余各时间点UHMVLs降低高尿酸血症大鼠模型血清中尿酸水平的效果均较游离UC更显著(P＜0.05).结论 在相同条件下,UHMVLs不仅能提高UC的活性,还可以增强UC的稳定性;UHMVLs在大鼠体内降低血尿酸水平的能力优于游离UC.     

浓烟吸入诱发大鼠透明质酸升高的研究

目的　探讨大鼠烟雾吸入后透明质酸增多与大鼠肺吸入性损伤的关系。方法　４０ 只ＳＤ 大鼠随机分成５ 组：正常对照组和烟雾吸入后不同时间观察组。分别测定血清、支气管肺泡灌洗液和肺组织中的透明质酸含量,支气管肺泡灌洗液中的炎性细胞比例及蛋白含量。结果　烟雾吸入后,血清和肺组织中的透明质酸含量升高,１２ ｈ 达到高峰［ 血清：１２ ｈ 组（４７０．４±９７ ．５） μｇ／Ｌ,对照组（１８６．４ ±３１．５） μｇ／Ｌ;肺组织：１２ ｈ 组（６９７ ．５±１１５．１）μｇ／Ｌ,对照组（２０２ ．６ ±３９．４） μｇ／Ｌ］ 。而支气管肺泡灌洗液的各时间观察组升高不显著。支气管肺泡灌洗液蛋白含量６ ｈ 达高峰;细胞数在１２ ｈ 最多,主要是中性粒细胞。结论　烟雾吸入后支气管内炎性细胞以中性粒细胞为主。肺内透明质酸含量高于血液。透明质酸似可作为烟雾吸入性肺损伤的病理指征。     

不同相对分子质量透明质酸对还原型谷胱甘肽透皮吸收的影响

研究不同相对分子质量透明质酸(HA)对还原型谷胱甘肽(GSH)在SD大鼠离体皮肤中透皮吸收和储留能力的影响.采用Franz扩散池法考察了不同相对分子质量HA对GSH的体外经皮渗透量及在不同层次皮肤中储留量的影响;利用分子对接AutoDock研究了 GSH与HA的相互作用;采用全反射傅里叶红外变换光谱(ATR-FTIR)...     

茵芍散对慢性乙肝湿热蕴脾证患者血清HA、IV-C水平的影响

观察茵芍散对慢性乙型肝炎湿热蕴脾证患者血清 HA、IV-C水平的影响。选择 64例慢性乙型肝炎湿热蕴脾证患者 ,随机分为 2组 ,分别在治疗前及治疗 1个月后观察各组上述指标变化。结果显示 :治疗组血清HA、IV-C含量明显降低 ,症候疗效明显改善 ,与对照组比较差异有显著性 (P<0 .0 5 )。说明茵芍散能改善血液循环 ,具有抗肝纤维化作用     

乙型肝炎肝硬化患者血清透明质酸与HBV-DNA的相关性研究

目的探讨乙肝肝硬化患者血清透明质酸与HBV-DNA水平的相关性。方法收集144例乙肝肝硬化患者,其中女性42例,男性102例,年龄（54．42±11．53）岁,Childpugh A级42例,ChildpughB级40例,Childpugh C级62例,采用放射免疫法检测血清透明质酸,荧光定量PCR检测血清HBV-DNA水平,对血清透明质酸、血清HBV-DNA水平进行统计并分析二者之间的关系。结果①随着乙肝肝硬化Child分级的加重,患者的血清透明质酸水平也增高,不同分级之间的差异有统计学意义（P〈0．01）。②不同Childpugh分级患者的血清HBV-DNA水平无显著的统计学意义（P〉O．05）。③不同Childpugh分级患者的血清HBV-DNA和透明质酸水平无显著相关性（P〉0．05）。结论随着患者的Childpugh分级升高,HA水平显著增高,说明HA是反映肝硬化程度的敏感指标;乙肝肝硬化患者HBV-DNA与肝硬化程度无显著相关性,血清透明质酸与HBV-DNA水平之间也无显著相关性,因此抗病毒和抗纤维化治疗对乙肝肝硬化患者同等重要。     

Instrumental,clinical and subjective evaluation of the efficacy of a cosmetic treatment for home use

Background: In the last few years, there has been increasing demand for aesthetic procedures to improve the effects of skin aging.

Aim: To evaluate the anti-aging efficacy, tolerability and skin changes induced by the topical products containing hyaluronic acid, N-acetyl glucosamine and gamma-amino butyric acid through instrumental techniques, clinical and subjective evaluation.

Patients/Method: Twenty female enrolled applied a day and night cream after applying a serum, once week applied a mask, for 2 months. A clinical assessment of smoothness, expression wrinkles, fine wrinkles and measurements of the parameters using Reveal Imager, X-Rite, Corneometer, Dermalab, Moisture Meter EpiD were taken at day 0, 15, 30 and 60 of study period. A final assessment questionnaire was submitted.

Results: The products were accepted by all the volunteers. The hydration (Corneometer: T0 49.17 vs T60 61.11, average variation 24.28%) (Moisture Meter EpiD: T0 45.73 vs T60 61.10, average variation 33.60%), elasticity (Dermalab: T0 56.06 vs T60 62.78, average variation 11.98%) and lightening of the skin (X-Rite: T0 60.23 vs T60 63.36, average variation 5.26%) improved. All changes were statistically significant (p < 0.05).

Conclusion: The efficacy of the topical products is confirmed by subjective, clinical and instrumental assessment. This should be a routine approach in dermatologic practice.