1.Simultaneous determination of the contents of 7 components in the Embelia laeta (L.) Mez with HPLC
Yi ZHANG ; Sha HAN ; Maochun HUANG ; Mushui XIE ; Huanxin ZHONG ; Zhiqiang GONG
International Journal of Traditional Chinese Medicine 2025;47(12):1739-1743
Objective:To establish an HPLC method to simultaneously determine the contents of 7 components, including Cardol triene, Cardol diene, (8'Z,11'Z,14'Z)-5-(Heptadeca-8',11',14'-trienyl)benzene-1,3-diol, Alkylresorcinol B, 5-(8'Z,11'Z-heptadecadienyl)-1,3-benzenediol, Adipostatin A, and 5-(11'Z-heptadecenyl)-resorcinol in Embelia laeta (L.) Mez. Methods:Chromatographic column was Arcus EP-C18(4.6 mm×250 mm, 5 μm); mobile phase was methanol-water (90∶10); column temperature was 25 ℃; flow rate was 1 ml/min; detection wavelength was 220 nm; the injection volume was 10 μl.Results:The 7 compounds in Embelia laeta (L.) Mez showed good linear relationships, which sample recovery rate ranges from 97.69%- 100.62%. The average contents of the 7 components from 9 origins were 3.099 9, 6.246 3, 9.942 8, 4.093 7, 2.180 3, 0.960 2, 1.855 9 mg/g. Conclusion:The established HPLC method for simultaneous determination of the contents of 7 components in Embelia laeta (L.) Mez is feasible, which can provide references for further development and utilization of Embelia laeta (L.) Mez.
2.Effect of different intervention plans on the expression of adhesion related cytokines and clinical outcomes after uterine adhesions separation surgery
Fen ZHAO ; Huanxin ZHONG ; Lifang YOU ; Yi LU ; Hongyin CUI
Chinese Journal of Endocrine Surgery 2024;18(5):739-743
Objective:To observe the effects of different intervention schemes on the expression of adhesion-related cytokines, menstrual recovery and clinical outcome of patients after transcervical resection of adhesion (TCRA) .Methods:180 patients received TCRA in our hospital from Feb. 2022 to Feb. 2023 were divided into group A, group B and group C according to different post-operative intervention programs, with 60 patients in each group. Patients in group A were treated with artificial cycle of estrogen and progesterone after surgery. On this basis, patients in group B were placed with a uterine birth control ring, and patients in group C were injected with sodium hyaluronate gel into the uterine cavity. The grade of uterine cavity adhesion, improvement rate of menstruation and pregnancy outcome at 2 months after operation and pregnancy outcome within 1 year after surgery were compared between the three groups at 2 months after operation. The relative mRNA expression of endometrial tissue transforming growth factor β1 (TGF-β1), platelet-derived growth factor BB (PDGF-BB), tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) and basic fibroblast growth factor (bFGF) at uterine cavity adhesion in each group were detected and compared.Results:At 2 months after surgery, the uterine adhesion rates in group A, group B, and group C were 43.33%, 15.00%, and 11.67%, respectively. There was no significant difference in the uterine adhesion rates between group B and group C ( P>0.05), but they were significantly lower than those in group A ( P<0.05) ; Meanwhile, the degree of intrauterine adhesions in group B and group C was significantly milder than that in group A ( P<0.05). The menstrual improvement rates of group A, group B, and group C at 2 months after surgery were 76.67%, 93.33%, and 96.67%, respectively. There was no significant difference between group B and group C ( P>0.05), but they were all significantly higher than group A ( P<0.05). At 2 months post surgery, the relative expression levels of TGF-β1, PDGF-BB, TIMP-1, and bFGF mRNA in the endometrial tissue at the site of uterine adhesions in group A were 0.77±0.26, 0.58±0.27, 0.54±0.15, and 0.62±0.14, respectively. In group B, they were 0.37±0.16, 0.37±0.14, 0.26±0.11, and 0.29±0.10, respectively. In group C, they were 0.32±0.16, 0.21±0.09, 0.27±0.08, and 0.34±0.18, respectively. The relative expression levels of cytokines in each group were significantly lower than during surgery ( P<0.05). There was no significant difference in the relative expression levels of various cytokines mRNA between group B and group C at 2 months after surgery ( P>0.05), but both were significantly lower than group A ( P<0.05). The pregnancy success rates within 1 year after surgery in group A, group B, and group C were 40.00%, 55.00%, and 58.33%, respectively. The pregnancy success rate in group C was significantly higher than that in group A ( P<0.05) . Conclusions:The application of metauterine contraceptive ring or sodium hyaluronate gel on the basis of estrogen and progesterone treatment after TCRA can effectively prevent postoperative re-adhesion of patients with intrauterine adhesions, improve clinical symptoms, and reduce the expression level of adhesion cytokines. The effects of the two schemes are equivalent.
3.N 6-methyladenosine-dependent pri-miR-17-92 mature activates AKT/mTOR pathway to promote endometrial cancer progression
Xiaoyan WANG ; Hongyin CUI ; Qingwen XIE ; Xiaoqian ZHOU ; Huanxin ZHONG
Chinese Journal of Endocrine Surgery 2022;16(6):698-702
Objective:To explore the role of N 6-methyladenosine (m6A) and its regulator METTL3 in the non-coding RNA of endometrial cancer.Methods:The expression levels of m6A and METTL3 were quantified in 20 paired carcinoma and adjacent clinical tissue samples from patients at from Jul. 2016 to Dec. 2020. HEC-1-A cell lines were constructed with METTL3 overexpression and knockdown. Western blot was used to detect the phosphorylation levels of key molecules in METTL3 and Akt/mTOR. The quantitative detection of mRNA levels were used qRT-PCR. The binding level of m6A to its receptor DGCR8 was determined by RNA immunoprecipitation.Results:The results of the m6A RNA methylation quantification kit showed that m6A (1.0±0.15) vs (1.7±0.34) ( P<0.01) and METTL3 levels were elevated in endometrial cancer cells, and METTL3 (1.0±0.13) vs (2.5±0.45) ( P<0.05) levels were elevated in endometrial cancer cells. Western blot and qRT-PCR detection of miR-17-92 cell clusters overexpressing METTL3, METTL3 overexpression significantly increased m6A modification on pri-miR-17-92 ( P<0.05) . Phosphorylation levels of AKT/mTOR pathway-related proteins were upregulated. In addition, RIP test results indicated that the binding of DGCR8 to pri-miR-17-92 was significantly facilitated. Conclusion:METTL3 modification of m6A facilitates the processing of pri-miR-1792 into the miR-17-92 clusters via m6A/DGCR8-dependent mechanism, which in turn activated the AKT/mTOR pathway.

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