RESUMEN
ObjectiveTo investigate the effect of betulinic acid （BA） on apoptosis and autophagy of human colorectal cancer SW620 cells and the regulatory role of phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt）/mammalian target of rapamycin （mTOR） signaling pathway. MethodCell viability was detected by methyl thiazolyl tetrazolium （MTT） colorimetry to determine the optimal administration time and dosage for subsequent experiments. Four groups were designed， including blank group and low-， medium-， and high-dose BA groups. Hematoxylin-eosin （HE） staining was conducted for the observation of SW620 cell morphology， and annexin-V/propidium iodide double staining for the determination of apoptosis rate in SW620 cells. Hoechst33258 staining and MDC staining were used for the observation of apoptosis and autophagy， respectively. Western blotting was employed to determine the protein levels of B-cell lymphoma/leukemia-2（Bcl-2）-associated X protein （Bax）， aspartate proteolytic enzyme-9 （Caspase-9）， activated aspartate proteolytic enzyme-3 （cleaved Caspase-3）， microtubule-associated protein 1 light chain 3 （LC3）， the mammalian homolog of yeast Atg6 （Beclin-1）， p62， phosphorylated PI3K （p-PI3K）， phosphorylated Akt （p-Akt）， and phosphorylated mTOR （p-mTOR） in SW620 cells. ResultBA inhibited the activity of SW620， HT29， and HCT116 cells in a concentration- and time-dependent manner. The cells treated with BA for 48 h had lower viability than those treated for 24 h （P<0.05， P<0.01）. The half maximal inhibitory concentration (IC50） value of BA at the time point of 48 h was also lower than that at the time point of 24 h （P<0.01）， and that for SW620 cells was the minimum. BA induced the apoptosis in a concentration-dependent manner and increased the autophagosomes. Compared with the blank group， BA increased the apoptosis rate （P<0.01）， up-regulated the protein levels of Bax， Caspase-9， cleaved Caspase-3， and LC3 Ⅱ （P<0.05， P<0.01）， and down-regulated the protein levels of p62， p-Akt， p-PI3K， and p-mTOR （P<0.01）. Additionally， medium- and high-dose BA up-regulated the protein level of beclin-1 （P<0.01）. ConclusionBA may inhibit the activity of SW620 cells by hindering the PI3K/Akt/mTOR signaling pathway to induce cell apoptosis and autophagy.
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This paper analyzes the registration regulations and guidelines of Traditional Chinese Medicine (TCM) products in South Africa, summarizes the registration path of TCM products in South Africa, analyzes the Common Technical Document (CTD) data required for registration from the aspects of quality control, safety and effectiveness, and discusses the registration strategy of TCM products in South Africa. This paper suggests that the strategies could include effective management and control of the registration risk of TCM and steady promotion of the products; TCM enterprises can first register low-risk TCM products, then open the South African TCM market, and promote the registration of high-risk TCM products after accumulating some experience; TCM enterprises need to have the awareness of promoting the inheritance, innovation and development of TCM, increase the investment in clinical trials of TCM products, and supplement the clinical effectiveness and safety data of TCM products; Strengthen the quality control of TCM and build an international brand of TCM.
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Ghana attaches great importance to the use of traditional medicine. Over the years, Ghana has successively promulgated laws and policies on traditional medicine, which has laid a legal foundation for the research, registration and sales of its herbal products. This article briefly describes the regulation system of herbal and food supplements in Ghana, and sorts out the registration path of Traditional Chinese Medicine (TCM) products as herbal and food supplements. We also briefly analyze the registration information. We believe that TCM products can open the Ghanaian market as food supplements, so as to promote the registration of herbal medicines. TCM enterprise should controlling TCM registration risks, and strengthening the cooperation between the Chinese and Ghana governments. TCM enterprise can leveraging on the advantages of TCM theory and experience, and strengthening cooperation with Ghana's traditional medicine. This article provide advices for the registration and listing of TCM products in Ghana, expanding the market of TCM in Ghana which finally expands to West Africa and the entire African countries.