1.Youguiwan Promotes Localization and Differentiation of Embryonic Stem Cells in Mouse Ovaries via Akt/mTOR/4EBP1/P70S6K Signaling Pathway
Xiang HU ; Mingya ZHU ; Qin ZHOU ; Jiayi SUN ; Zimo LI ; Xiao WANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(22):51-60
ObjectiveTo observe the effects of Youguiwan on the localization and differentiation of embryonic stem cells transplanted into ovaries,and to preliminarily explore whether its mechanism is related to the protein kinase B (Akt)/mammalian target of rapamycin (mTOR)/4E-binding protein 1 (4EBP1)/p70S6 kinase (P70S6K) signaling pathway. MethodsA total of 81 healthy adult female C57BL/6 mice that had received ovarian transplantation of GFP-labeled embryonic stem cells were randomly allocated into a control group,a Youguiwan group,and a positive control group. The control group received normal saline via gavage,the Youguiwan group was administered Youguiwan decoction at a clinical equivalent dose of 17.1 g·kg-1,and the positive control group received human menopausal gonadotropin (HMG) injections every 48 h. Ovarian tissue samples were collected on days 7,14,and 21 post-administration. Frozen sections were prepared to observe stem cell localization. The immunofluorescence assay was used to detect growth differentiation factor 9 (GDF9) and mTOR expression. real time polymerase chain reaction(Real-time PCR) was performed to measure the mRNA levels of follicle-stimulating hormone receptor (FSHR) and luteinizing hormone receptor (LHR). Western blotting was conducted to assess the expression and phosphorylation levels of Akt,mTOR,4EBP1,and P70S6K. ResultsFluorescence microscopy revealed that the transplanted embryonic stem cells were predominantly localized within the granular cell layer,the cell layer of the follicular structure,and the stroma. After 14 days of drug intervention,compared with the control group,the Youguiwan group showed upregulated mRNA levels of FSHR and LHR in the ovarian tissue (P<0.01),as well as elevated phosphorylation/protein ratios of Akt,mTOR,4EBP1,and P70S6K (P<0.05). After 21 days of drug intervention,compared with the control group,the Youguiwan group exhibited enhanced expression of GDF9 and mTOR in the ovarian tissue as detected by immunofluorescence (P<0.05),along with elevated mRNA levels of FSHR and LHR (P<0.01) and phosphorylation/protein ratios of Akt and 4EBP1 (P<0.05). ConclusionYouguiwan can promote the localization and differentiation of mouse embryonic stem cells within the ovary and enhance the expression of GDF9,FSHR,and LHR mRNA in the ovarian tissue by influencing the Akt/mTOR/4EBP1/P70S6K pathway.
2.THE INFLUENCE OF DIFFERENT CULTURALCONDITIONS TO THE OOSPORE FORMATION
Jiehua ZHU ; Zhiming ZHANG ; Mingya DING ; Yuqin LI ; Junyu YANG
Microbiology 1992;0(05):-
This paper dealt with the influences of different media, temperature, light and pH to the amount of oospore formation. The results showed that millet agar, bai yun beau agar and soybean agar provided the favorable condition for oospore formation, the number of oospores was 131.6~149.6 /cm2, but a few oospores was formation on frozen pea agar. The results also suggested that the optimal conditions for oospore formation were 18℃, pH7 and darkness. The oospores failed to be formed tinder fluorescent or black light.

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