Objective To establish a human luminal breast cancer stem cell(BCSC)model and investigate the inhibitory effects of astragaloside Ⅳ(AS-Ⅳ)on BCSC growth.Methods MCF-7 breast cancer cells were cultured in stem cell-specific medium to induce BCSC formation.The BCSCs were then divided into a blank control group and an AS-Ⅳ treatment group,both groups were given PBS or AS-Ⅳ treatment.Morphological changes were observed after intervention.The therapeutic efficacy of AS-Ⅳ was evaluated using 3D spheroid formation and cell viability assays.Transcriptomic profiling and gene expression analysis were performed to elucidate the underlying mechanisms.Results Compared with the MCF7 breast cancer cells,MCF7 breast cancer stem cell mammospheres exhibited accelerated growth(P＜0.01)and significantly increased expression of the stemness marker ALDH1A1(P＜0.01).Further comparison with the blank control group revealed that astragaloside Ⅳ(AS-Ⅳ)treatment significantly inhibited MCF7 breast cancer stem cell proliferation(P＜0.001)and slowed mammosphere growth(P＜0.01).Transcriptomic analysis demonstrated that differentially expressed genes(DEGs)induced by stem cell modeling and AS-Ⅳ intervention were enriched in the cellular senescence signaling pathway.AS-Ⅳ intervention substantially increased the number of SA-β-gal-positive cells(P＜0.01).RT-PCR analysis confirmed that AS-Ⅳsignificantly upregulated mRNA expression of IL-1α(P＜0.01),P21(P＜0.001),and P53(P＜0.05)in MCF7 breast cancer stem cells.Conclusion Astragaloside Ⅳ suppresses the growth of human luminal breast cancer stem cells by inducing cellular senescence.

Objective To evaluate the effect of exosomes from different sources on the growth, metastasis and immune microenvironment of hepatocellular carcinoma ( HCC ) in a mouse model of orthotopic transplanted hepatoma so as to provide new insight into the clinical immunotherapy of hepatocellular carcinoma. Methods The serum-derived exosomes were obtained by ultracentrifugation from Hepa1-6 cells and 3-week orthotopically tumor-bearing HCC mice. The morphology and size of exosomes were identified by transmission electron microscopy. One-week tumor-bearing HCC C57BL/6 mice were randomly divided into 3 groups ( 3 mice for each group ) , and respectively treated with tail vein injection of 120 μl PBS(control group), 120 μl Hepa1-6 cell-derived exosomes (1μg/μl, TEXcel group), and serum-derived exosomes (1μg/μl) (TEXserum group). The treatments were conduced once a week for 3 consecutive weeks. The mice were sacrificed on the 4th day after the treatments, and the liver tissue and lung tissues were dissected. The volumes of the liver cancer tissues were measured. The expression of PD-L1 protein and CD3+and FoxP3+T lymphocytes infiltration in the liver cancer tissues were respectively detected by Western Blot and immunohistochemical staining. The lung metastasis of the liver cancer was detected by hematoxylin-eosin staining. Results The diameters of the Hepa1-6 cell- and serum-derived exosomes both were about 100 nm, and were uniform vesicles with complete membrane structure. Compared with the control group, Hepa1-6 cell-derived exosomes had a certain inhibitory effect on the growth and lung metastasis of HCC, while there was no significant difference for the TEXserum group. Western Blot results showed that compared with the control group, PD-L1 protein expression was significantly up-regulated in both TEXcel group and TEXserum group. Immunohistochemical staining showed that compared with the control group, the infiltration number of Foxp3+-labeled regulatory T cells (Treg) had no significant changes in TEXcel group and the TEXserum group, and the differences were no statistically significant (all P>0.05). However, the infiltration number of CD3+-labeled T lymphocytes was significantly reduced in the two groups, and the differences were statistically significant (all P<0.05). Conclusion The Hepa1-6 cell-derived exosomes have a certain inhibitory effect on the growth and lung metastasis of HCC, and have no obvious regulation effects on the immune microenvironment of HCC. The serum-derived exosomes from orthotopically tumor-bearing HCC mice can promote the growth and lung metastasis of HCC and immunosuppress the microenvironment. The Hepa1-6 cell-derived exosomes are expected to be used for immunotherapy studies of liver cancer.

Objective To establish the gas chromatographic(GC) fingerprint of oleic acid of Whitmania pigra Whitman for its quality control. Methods Ten batches of Whitmania pigra from different sources and processed by different methods were analyzed with Agilent 6890N gas chromatography detector on DB-WAX(30 mm × 0.32 mm × 0.25μm)column at the vaporizing temperature of 270℃, column temperature of 130℃and flame ionization detector (FID) temperature of 280℃. We used a software of Similarity Evaluation System for Chromatographic Fingerprint of TCM(Version of 2004A) published by Chinese Pharmacopoeia Commission to calculate GC similarity. Results Oleic acid content of Whitmania pigra processed by different methods had significant differences (F2,7 = 7.350, P = 0.019). The oleic acid content of samples dried after washing with clean water significantly differed from that of the samples processed by alumen or the slices dried naturally(P = 0.021, P= 0.009). The similarity of the fingerprints was in the range of 0.458 - 0.998. The similarity of samples from Lipu of Guangxi Province was the lowest. Conclusion The fingerprints of most of the samples have very high similarity. The established GC fingerprints can be used to effectively identify the qualified or inferior Whitmania pigra products, which will provide some reference for the quality control of Whitmania pigra.

Objective To explore the effect of α-zearalanol (α-ZAL) on β-amyloid (Aβ) induced mice and the mechanism. Methods The model was induced by intracerebroventricular injection of Aβ25-35. The mice were divided randomly into sham group, model group, estradiol benzoate (EB) group (Aβ+EB) as a positive control and α-ZAL (Aβ+α-ZAL) group. Morris water maze was used to evaluate the learning and memory ability. The levels of antioxidant enzymes and nitric oxide system in the brain tissue were detected with spectrophotometric and sotopic method. Results The escape latency was longer in the model group than in the control group (P<0.01), and was shorter in the EB group and α-ZAL group than in the model group (P<0.05). Compared with the control group, the level of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) decreased, and the level of malonaldehyde (MDA), constitutive nitric oxide synthase (cNOS), inducible nitrous oxide synthesis (iNOS), and nitric oxide (NO) increased in the model group (P<0.05); compared with the model group, the level of SOD and GSH-Px increased, and the level of MDA, cNOS, iNOS and NO decreased in the EB group and α-ZAL group (P<0.05), except the level of SOD and cNOS in hippocampus in α-ZAL group (P>0.05). There was no significant difference between EB group and α-ZAL group (P>0.05). Conclusion α-ZAL could improve the cognitive behavior in Aβ25- 35 induced mice by increasing the antioxidant activities and decreasing the lipid peroxidation.

Traditional Chinese medicine (TCM) have unique efficacy and advantages for treating diseases of central nervous system. In recent years, some researches focused on neural stem cells and neurogenesis. The researches about TCM for influencing neural stem cells biological characteristics and common neurological diseases were reviewed.