Examination of the correlation between grades, chemicalcharacteristics and anti-fatigue effect of Astragali Radix / 药学学报

To provide a basis for the establishment of the commodity grade of Astragali Radix (AR), we compared the chemical components and the anti-fatigue effect of different grades of AR. The components of primary metabolites were analyzed by 1H NMR and the contents of five flavonoids were determined by HPLC-UV with different grades of AR. Fatigue efficacy of different grades of AR was compared. All the procedures were approved by the Laboratory Animal Ethics Committee of the Shanxi University. The results showed that the content of water soluble extracts (WSE) of the Grade Ⅱ AR was the lowest, and 21 compounds were identified through 1H NMR spectrum. There are 3 components showing a higher content in the Grade-top AR, and 7 components were higher in the Grade-Ⅳ, and 7 other components were higher in the Grade-Ⅱ AR. Total flavonoid content was the highest in Grade-Ⅱ but it was the lowest in the Grade-Ⅳ. Pharmacodynamic results showed that AR could significantly enhance the exhaustion time of rats and improve the biochemical indexes of serum and gastrocnemius muscle, and the best anti-fatigue effect was observed with Grade -Ⅱ AR. Therefore, chemical composition and efficacy index were used to evaluate the quality of different grades of AR, and the quality evaluation approach was established based on chemical and pharmacological effects to provide a scientific basis for the development of AR. The study may provide useful information for construction of the quality grade standard of AR.

Optimization of determination methods of astragaloside Ⅳ and analysis of contents of astragaloside Ⅳ in different origin and grade / 中国中药杂志

A rapid and accurate method for determination of astragaloside Ⅳ was established,which was further applied to determine the contents of astragaloside Ⅳ in 87 batches of different origin and different grade of Astragali Radix. The ROC curve was used to analyze the contents of astragaloside Ⅳ in different origin. Simultaneous contents of astragaloside Ⅳ in different grade were compared with chemometrics. HPLC-ELSD method was used to determine the contents of astragaloside Ⅳ. A Vensil MP C18 column( 4. 6 mm×250 mm,5 μm) was used with acetonitrile-water( 32 ∶68) as the mobile phase at a flow rateof 1 m L·min-1. The column temperature was 25 ℃ with ELSD parameters as follows: gas flow rate was 2. 5 L·min-1,the drift tube heating temperature was set to 105 ℃,and the gain value was 4. 0. The optimized method avoided the problem that the consumable quality unstable and the recovery rate was not high. The contents determined by the optimized method were higher than the pharmacopoeia method,with less time and high recovery rate. The ROC curve analysis showed that there was no significant difference of contents of astragaloside Ⅳ between the top grade of Shanxi wild-simulated Astragali Radix top and the first grade of Gansu cultivated Astragali Radix. The contents of astragaloside Ⅳ in the second,third and fourth grade of Shanxi wild-simulated Astragali Radix was significantly higher than those of produced from Gansu.There was a significant negative correlation between the contents of astragaloside Ⅳ and grade in Shanxi Astragali Radix. While there was no correlation for Gansu Astragali Radix. This study provided the basis for the quality grade standard of Astragali Radix.

Determination of Content Limit of Isoflavone Glucoside of Hengshan Wild-simulated Astragali Radix by ROC Curve Method / 中国实验方剂学杂志

Objective: To explore the receiuer operating characteristic(ROC) curve mathematical statistics method to establish the content limit of isoflavones glucoside contents in Astragali Radix,in order to distinguish the difference in isoflavones glucoside content between Hengshan wild-simulated Astragali Radix in Shanxi and cultivated Astragali Radix in Gansu. Method: A total of 225 samples of 45 batches of wild-simulated Astragali Radix in Hengshan and 210 samples of 42 batches of cultivated Astragali Radix in Gansu were selected as research objects,and isoflavones glucoside contents of these samples were determined by using the method in the edition 2015 Chinese Pharmacopoeia. Data was statistically analyzed, and the ROC curve was used to determine the critical value of wild-simulated Astragali Radix in Hengshan and transplanted Astragali Radix in Gansu. Result: The average contents of isoflavones glucoside in imitative wild Astragali Radix in Henshan and cultivated Astragali Radix in Gansu were 0.107%and 0.039%respectively,with statistically significant differences(PConclusion: ROC curve can be used to distinguish the critical value of the wild-simulated Astragali Radix in Hengshan and cultivated Astragali Radix in Gansu. The results reflected the regularity of "high quality and good price" in the market. The research provided a theoretical basis for the establishment of standard of high-quality Chinese herbal medicines.

Tumor-infiltrating regulatory T cells are positively correlated with angiogenic status in renal cell carcinoma / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Immune cells within a tumor microenvironment have shown modulatory effects on tumor angiogenic activity. Renal cell carcinoma (RCC) is a hypervascular tumor that reportedly increases the frequency of regulatory T cells (Tregs) in tumor tissues. This study investigated the correlation between Tregs infiltration and angiogenic status in RCC.</p><p><b>METHODS</b>Thirty-six patients with RCC were enrolled in the present study, and twenty age-matched healthy donors were included as the control. Tregs were defined as CD4(+)CD25(high)CD127(low/-) T cells. The frequency of Tregs in peripheral blood and tumor infiltrating lymphocytes (TILs) were determined by flow cytometry. The expression of vascular endothelial growth factor (VEGF) in surgical resection specimens were measured with a commercial enzyme-linked immunosorbent assay (ELISA) kit. Microvessel density (MVD) was calculated on slides stained with CD34 antibody. Spearman's rank correlation was performed to evaluate the correlation between the frequencies of Tregs in TILs and VEGF values, as well as between frequencies of Tregs and MVD determinations.</p><p><b>RESULTS</b>Compared to healthy controls, the frequency of peripheral blood Tregs was significantly increased in patients with RCC (P < 0.05). The percentage of tumor-infiltrating Tregs was higher than that of peripheral blood Tregs in patients with RCC (P < 0.01). In addition, the frequency of tumor-infiltrating Tregs was shown to significantly correlate with the pathological stage (P < 0.05) and nuclear grade (P < 0.01). Importantly, a significant positive correlation was observed between the frequency of tumor-infiltrating Tregs and VEGF protein expression (r = 0.51, P < 0.05), as well as between frequencies of Tregs and MVD score (r = 0.39, P < 0.05).</p><p><b>CONCLUSIONS</b>These observations suggest that the high pro-angiogenic status of RCC may be associated with the accumulation of Tregs in the local microenvironment. Angiogenesis networks may be connected with immune tolerance units and cooperate with each other to facilitate tumor growth and progression.</p>

An analysis for the phenotype and genotype of autosomal dominant polycystic kidney disease from two Chinese families / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To scan for mutations of polycystic kidney disease 1 gene (PKD1) in Chinese population in order to find some features about Chinese patients and a better approach to detect mutations.</p><p><b>METHODS</b>Twenty-five PKD-affected individuals from twenty-one unrelated genealogies and sixteen controls participated in the study. Thirty-five blood samples and six tissues were obtained after receiving informed consent and were in accordance with institutional ethical guidelines. Genomic DNA was isolated from peripheral blood using standard procedures. PCR amplification of genomic DNA was performed to generate the aimed fragments. Amplified fragments were analyzed by denaturing gradient gel electrophoresis (DGGE). A GC clamp was attached to the 5' primer. After that, the abnormal fragments were sequenced on freshly amplified specific PCR products with the dideoxynucleotide chain termination method. Sequencing was performed for all samples to evaluate DGGE.</p><p><b>RESULTS</b>Aimed fragments of exons 44 and 45 were amplified. DGGE detected eleven abnormal PCR fragments. Two novel mutations were identified by sequencing, included one nonsense mutation (C12217T) and one frameshift (12431delCT). In addition, one polymorphism (A50747C) was identified. The mutation detection rate is 8% in our study.</p><p><b>CONCLUSION</b>Two novel pathogenic mutations were detected, including one nonsense mutation (C12217T) and one frameshift (12431delCT).</p>