Resveratrol, an Inhibitor Binding to VEGF, Restores the Pathology of Abnormal Angiogenesis in Retinopathy of Prematurity (ROP) in Mice: Application by Intravitreal and Topical Instillation.

Retinopathy of prematurity (ROP) is a severe eye disease leading to blindness. Abnormal vessel formation is the pathological hallmark of neovascular ROP. In forming vessels, vascular endothelial growth factor (VEGF) is an important stimulator. The current anti-ROP therapy has focused on bevacizumab, a monoclonal antibody against VEGF, and pazopanib, a tyrosine kinase inhibitor on the VEGF receptor (VEGFR). Several lines of evidence have proposed that natural compounds may be more effective and safer for anti-VEGF function. Resveratrol, a common natural compound, binds to VEGF and blocks its interaction with VEGFR, thereafter suppressing angiogenesis. Here, we evaluate the efficacy of intravitreal injection, or topical instillation (eye drops), of resveratrol into the eyes of mice suffering from oxygen-induced retinopathy, i.e., developing ROP. The treatment of resveratrol significantly relieved the degree of vascular distortion, permeability and hyperplasia; the efficacy could be revealed by both methods of resveratrol application. In parallel, the treatments of resveratrol inhibited the retinal expressions of VEGF, VEGFR and CD31. Moreover, the applied resveratrol significantly relieved the damage caused by oxygen radicals through upregulating the level of superoxide dismutase (SOD) and downregulating the level of malondialdehyde (MDA) in the retina. Taken together, the potential therapeutic benefit of resveratrol in pro-angiogenic diseases, including retinopathy, can be considered.

[Determination of Nucleosides and HPLC Fingerprints of Cordyceps].

OBJECTIVE: To establish the HPLC fingerprints method of Cordyceps and to determine the contents of uridine, inosine, guanosine and adenosine. METHODS: The HPLC separation was performed on a Grace Prevail C18 column( 150 mm x 4.6 mm, 5 µm) in a gradient elution mode with a mixture consisting of water and methanol at a flow rate of 1.0 mL/min, the detection wavelength was set at 260 nm, the column temperature was 25 degrees C. RESULTS: The contents of four nucleosides were determined in Cordyceps from different habitats, and the HPLC fingerprint of Cordyceps was set up with 13 common peaks. Among of them, uridine, inosine, guanosine and adenosine were identified. The similarities of ten fingerprints were greater than 0.95 with good separation of each chromatographic peak, and met the requirement of the fingerprints. There were similar results in cluster analysis and principal component analysis of the major nucleosides and the fingerprints of 10 batches of Cordyceps. The results of sample classification in principal component analysis showed a good similarity with cluster analysis. CONCLUSION: This method showed the information of chemical composition in Cordyceps, with good repeatability and similarity between samples, indicating that the stable chemical distribution and proportion of the major nucleosides in the medical materials. Fingerprints, principal component analysis and cluster analysis, which are applied to identify the different sources of Cordyceps, provide an experimental basis for establishing the characteristics evaluation methodology of medicinal materials.

[Determination of morusin in cortex mori].

OBJECTIVE: To determine the contents of morusin in Sangbaipi (Cortex Mori) of different commercial grades and species and obtained from different regions. METHOD: Contents of morusin were determined by HPLC on an Alltima C18 column (4.6 mm x 250 mm, 5 microm) with acetonitrile-water (58:42) as mobile phase. UV detection was conducted at 270 nm. RESULT: The amounts of morusin were found to vary among samples of Cortex Mori bearing a yellow-brown outer layer and those without the yellow-brown cork layer. The morusin contents were also related to the thickness of Cortex Mori. CONCLUSION: The method was simple, reproducible and reliable. It can be applied to the quality assessment of Cortex Mori.

[Determination of heavy metals in four traditional Chinese medicines by ICP-MS].

OBJECTIVE: To establish a ICP-MS method for the determination of heavy metals, including As, Hg, Pb, Cd, in four traditional Chinese medicines. METHOD: The samples were digested by closed-versel microwave. The four heavy metals were directly analyzed by ICP-MS. Select internal standard element in for the method by which the analyse signal drife is corrected by the signal of another element (internal standard elements) added to both the standard solution and sample. RESULT: For all of the analyzed heary methals, the correlative coefficient of the calibration curves was over 0.999 2. The recovery rates of the procedure were 97.5%-108.0%, and its RSD was lower than 11.6%. CONCLUSION: This method was convenient, quick-acquired, accurate and highly sensitive. The method can be used for the quality control of trace elements in traditional Chinese medicines and for the contents determination of traditional Chinese medicines from different habitats and species.

[The fingerprint of Ezhu by GC-MS].

AIM: To study the fingerprint of Ezhu by GC-MS. METHODS: GC-MS analysis was performed for 18 samples of three species of Curcuma used as Ezhu. TIC profiles were evaluated by "Computer Aided Similarity Evaluation System" (MATLAB5.3 based, Ver. 1.240, developed by Research Center for Modernization of Chinese Medicine, Central South University). The characteristic peaks in chromatograms were identified by comparing mass data with literatures. Hierarchical clustering analysis was performed by SPSS based on the relative peak area (RPA) of identified peak to germacrone in 18 samples. RESULTS: Resemblance values of 18 samples of Ezhu were pretty low. The mutual mode fingerprint plots of Ezhu were failed to develop. However, 18 samples were divided into two main clusters based on hierarchical clustering analysis, Curcuma wenyujin cluster and Curcuma phaeocaulis cluster, but the samples of Curcuma kwangsiensis were dispersive. Therefore, based on hierarchical clustering analysis, two mutual mode fingerprint plots of Curcuma wenyujin and Curcuma phaeocaulis were developed. But that of Curcuma kwangsiensis was failed because of low resemblance among samples. CONCLUSION: The mutual mode fingerprint is the basis for quality control of Chinese materia medica from multi-origins. Development of GC-MS fingerprint of Ezhu was failed, which indicates that the chemical components in different species of herbs used as one Chinese materia medica may be significantly different. The relationship of chemical components and pharmacological activities should be further studied so as to elucidate the rationality of Chinese materia medica from multi-origins.