Objective To establish a method for the Determination of Danshensu content in Tongmai Capsules. Methods The content of Danshensu was determined by HPLC on Kromasil C18 Column( 4.6? 250 mm, 5 ? m) . The mobile phase was acetonitri1- 1.2 % water solution of acetic acid (9∶ 91) and the detection wavelength was 280 nm. The theoretical plates should over 3000 according to Danshensu. Results Sodium Danshensu showed a good linearity in the range of 0.288～ 1.152 ? g, r=0.9994. The average recovery was 97.62 % , and RSD was 1.10 % (n=5). Conclusion This method is effective and can be used for the quality control of Danshensu in Tongmai Capsules.

Objective To establish a method for the determination of hesperidin content in Qiansheng Baizhusan Jiawei Granule (QBJG). Methods The content of hesperidin was determined by HPLC. Results Hesperidin showed a good linearity in the range of 0. 22-1.1ug(r = 0. 9998). The average recovery was 103. 0 % and RSD was 3. 12 %. Conclusion The methods are accurate and can be used for the quality control of QBJG.

Objective: To establish the quality standard for Guangxiao Liniao Capsules (Herba Verbenae, Herba Leonuri, Rhizoma Alismatis, Radix Linderae, etc.). Methods: Herba Verbenae, Herba Leonuri, Rhizoma Alismatis, Radix Linderae were identified by TLC, and the content of stachydrine hydrochloride was determined by TLC scanning. Results: Herba Verbenae, Herba Leonuri, Rhizoma Alismatis, Radix Linderae could be identified by TLC. Stachydrine hydrochloride showed a good linear relationship at a range of 4?g～20?g, r = 0.9975. The average recovery was 97.9%, and RSD was 1.20%. Conclusion: The methods are accurate and can be used for the quality control of Guangxiao Liniao Capsules

AIM: To establish the quality standards for Fukang Capsules(Cortex Phellodendri Chinensis,Cortex Ailanthi,Fructus Schisandrae Chinensis,etc.). METHODS: Cortex Ailanthi,Fructus Schisandrae,Poria were identified by TLC,and the content of berberine hydrochloride was determined by TLC-scanning. RESULTS: Cortex Ailanthi,Fructus Schisandrae,Poria could be identified by TLC.Berberine hydrochloride showed a good linear relationship at a range of 25.32 ng-354.48 ng,r=0.993 28.The average recovery was 100.3%,and RSD was 2.04%. CONCLUSION: The method is accurate and can be used for the quality control of Fukang Capsules.

Objective: To establish a method for determinating icarin content in Naoling Granule. Methods: Water-soluble ethyl-acetate-extracting polyamide column purification was adopted. The content of icarin was determined by HPLC. C18 column was used, mobile phase was methanol:0.1mol/mL sodium phosphate dibasic (6:5) and detection wavelength was at 270nm. Results: Icarin has a good linearity in the range of 0.24～1.2?g ( r=0.9998). The average recovery was 98.23 %and RSD was 1.2 %(N=5). Conclusion: The method was simple with good reproducibility and can be used for the content determination of icarin in Naoling Granule.

Objective: To establish a method for the determination of gastrodine in Tianma Formula Granule. Methods: HPLC was used with LiChrospherR 100 column, MeOH-phosphate solution(contain KH2PO4 and Na2HPO4 each 0.1mol/L)-water(1.5:3:95.5) as mobile phase and detection wavelength at 270nm. Results: Gastrodine showed a good linearity in the range of 1.638～14.742?g. The average recovery was 98.17 %and RSD was 1.39 %(n=5). Conclusion: This method is simple, accurate and with good reproducibility, and can be used for the quality control of Tianma Formula Granule.

Objective To compare the effects of two crushing methods on dissolution rate and decocting rate of water-soluble protein in Plastrum Testudinis.Methods The dissolution rate of water-soluble protein was compared in Plastrum Testudinis processed by the traditional crushing method(passing 100-eyes screen,fine powder)and ultra-fine crushing method(passing 300-eye screen,ultra-fine powder);besides,orthogonal design was applied to study the process technical condition of decocting rate in Plastrum Testudinis.Results Water-soluble protein in fine powder of Plastrum Testudinis was hardly detected while the dissolution rate of water-soluble protein in ultra-fine powder was 51.2 %in 20 minutes and up to 70.5 %at 2 hours.Three influencing factors of fineness,decocting frequencies and decocting time were measured with orthogonal test.The results of variance analysis showed that fineness significantly influenced the decocting rate(P .05).Conclusion The ultra-fine powder technique is propitious to the increase of dissolution rate and decocting rate of water-soluble protein in Plastrum Testudinis.

Objective:To investigate the effect of miR-27b-3p on radiation resistance of breast cancer cells.Methods:The relative expression levels of miR-27b-3p in normal tissues and breast cancer tissues were analyzed through GEO database and verified by the qRT-PCR assay. Cloning formation, immunofluorescence, and EDU assay were used to assess the functions of miR-27b-3p on radioresistance of breast cancer cells. The luciferase reporter assay was used to verify whether miR-27b-3p directly targeted PLK2 mRNA. A rescue experiment was performed to identify the influence of PLK2 overexpression on miR-27b-3p regulated radioresistance.Results:The expression level of miR-27b-3p in both breast cancer tissues ( t=2.99, P<0.01) and breast cancer cells ( t=21.21, 32.88, P<0.05) was significantly higher than those in normal breast tissues and cells, especially, it was elevated in radioresistant MCF-7R cells ( t=25.63, P<0.05). Overexpression of miR-27b-3p enhanced the cloning efficiency of MCF-7 cells ( t=10.32, P<0.05), and had a protective effect on the proliferation of irradiated MCF-7 cells ( t=8.77, 8.26, 8.03, P<0.05). But interference of miR-27b-3p reduced the cloning efficiency and proliferation of MCF-7R cells ( t=40.00, P<0.05) after irradiation with different doses( t=8.54, 8.32, 8.23, P<0.05). Moreover, PLK2 was verified to be a direct target of miR-27b-3p, and overexpression of PLK2 inhibited miR-27b-3p-mediated radioresistance of breast cancer cells (MCF-7: t=9.66, P<0.05; MCF-7R: t=6.42, P<0.05). Conclusions:miR-27b-3p contributes to the radioresistance of breast cancer cells by targeting PLK2.

Cryoablation (CRA) and microwave ablation (MWA) are two main local treatments for hepatocellular carcinoma (HCC). However, which one is more curative and suitable for combining with immunotherapy is still controversial. Herein, CRA induced higher tumoral PD-L1 expression and more T cells infiltration, but less PD-L1^highCD11b⁺ myeloid cells infiltration than MWA in HCC. Furthermore, CRA had better curative effect than MWA for anti-PD-L1 combination therapy in mouse models. Mechanistically, anti-PD-L1 antibody facilitated infiltration of CD8⁺ T cells by enhancing the secretion of CXCL9 from cDC1 cells after CRA therapy. On the other hand, anti-PD-L1 antibody promoted the infiltration of NK cells to eliminate PD-L1^highCD11b⁺ myeloid cells by antibody-dependent cell-mediated cytotoxicity (ADCC) effect after CRA therapy. Both aspects relieved the immunosuppressive microenvironment after CRA therapy. Notably, the wild-type PD-L1 Avelumab (Bavencio), compared to the mutant PD-L1 atezolizumab (Tecentriq), was better at inducing the ADCC effect to target PD-L1^highCD11b⁺ myeloid cells. Collectively, our study uncovered the novel insights that CRA showed superior curative effect than MWA in combining with anti-PD-L1 antibody by strengthening CTL/NK cell immune responses, which provided a strong rationale for combining CRA and PD-L1 blockade in the clinical treatment for HCC.