Objective:To investigate whether intermittent fasting alleviates insulin resistance in a polycystic ovary syndrome(PCOS) mouse model through the regulation of autophagy.Methods:Fifty 3-week-old female C57BL/6J mice were randomly assigned into the following groups using a random number table: normal control(NC) group( n=10), maintained on a standard chow diet; high-fat diet(HFD) group( n=10) fed a diet with 60% of calories derived from fat; and PCOS model group( n=30), established by combining a HFD with dehydroepiandrosterone(DHEA) administration. Successful modeling was confirmed by disrupted estrous cycles, hyperandrogenism, and polycystic ovarian morphology. The PCOS model mice were further divided into three groups: PCOS group( n=9), PCOS with intermittent fasting group(PCOS+ IF, n=9), and PCOS with intermittent fasting plus the autophagy inhibitor 3-methyladenine(3-MA) group(PCOS+ IF+ 3-MA, n=9). Autophagy levels were assessed by detecting markers LC3 and p62 and observing autophagosomes via transmission electron microscopy. Glucose tolerance test(GTT) and insulin tolerance test(ITT) were performed, and the area under the curve(AUC) was calculated to evaluate insulin resistance. Western blotting was used to detect phosphorylation levels of phosphatidylinositol 3-kinase(PI3K), protein kinase B(Akt), mammalian target of rapamycin(mTOR), and p70S6 kiase(p70S6K). Results:Compared with the NC group, the PCOS model group showed absent estrous cycles, significantly elevated serum testosterone, sex hormone binding globulin, and luteinizing hormone(LH) levels( P<0.001), and polycystic ovarian changes on hematoxylin-eosin staining, confirming successful model establishment. Immunohistochemistry, transmission electron microscopy, and Western blotting demonstrated that autophagy levels were increased in the PCOS+ IF group compared with the PCOS group, while 3-MA administration reduced the intermittent fasting - induced autophagy. The AUC values for both GTT and ITT were significantly lower in the PCOS+ IF group than those in the PCOS group( P<0.001, P=0.003), but increased in the PCOS+ IF+ 3-MA group compared to the PCOS+ IF group( P<0.001, P=0.020). Western blotting analysis showed that phosphorylation levels of PI3K, Akt, mTOR, and p70S6K were significantly decreased in the PCOS+ IF group compared with the PCOS group( P=0.002, P=0.001, P=0.001, and P<0.001, respectively), and increased in the PCOS+ IF+ 3-MA group compared with the PCOS+ IF group( P=0.021, P=0.041, P=0.047, and P=0.024, respectively). Conclusions:Intermittent fasting alleviates insulin resistance in a PCOS mouse model through inhibitiing PI3K/Akt/mTOR signaling pathway and promoting autophagy.

Objective:To investigate whether intermittent fasting alleviates insulin resistance in a polycystic ovary syndrome(PCOS) mouse model through the regulation of autophagy.Methods:Fifty 3-week-old female C57BL/6J mice were randomly assigned into the following groups using a random number table: normal control(NC) group( n=10), maintained on a standard chow diet; high-fat diet(HFD) group( n=10) fed a diet with 60% of calories derived from fat; and PCOS model group( n=30), established by combining a HFD with dehydroepiandrosterone(DHEA) administration. Successful modeling was confirmed by disrupted estrous cycles, hyperandrogenism, and polycystic ovarian morphology. The PCOS model mice were further divided into three groups: PCOS group( n=9), PCOS with intermittent fasting group(PCOS+ IF, n=9), and PCOS with intermittent fasting plus the autophagy inhibitor 3-methyladenine(3-MA) group(PCOS+ IF+ 3-MA, n=9). Autophagy levels were assessed by detecting markers LC3 and p62 and observing autophagosomes via transmission electron microscopy. Glucose tolerance test(GTT) and insulin tolerance test(ITT) were performed, and the area under the curve(AUC) was calculated to evaluate insulin resistance. Western blotting was used to detect phosphorylation levels of phosphatidylinositol 3-kinase(PI3K), protein kinase B(Akt), mammalian target of rapamycin(mTOR), and p70S6 kiase(p70S6K). Results:Compared with the NC group, the PCOS model group showed absent estrous cycles, significantly elevated serum testosterone, sex hormone binding globulin, and luteinizing hormone(LH) levels( P<0.001), and polycystic ovarian changes on hematoxylin-eosin staining, confirming successful model establishment. Immunohistochemistry, transmission electron microscopy, and Western blotting demonstrated that autophagy levels were increased in the PCOS+ IF group compared with the PCOS group, while 3-MA administration reduced the intermittent fasting - induced autophagy. The AUC values for both GTT and ITT were significantly lower in the PCOS+ IF group than those in the PCOS group( P<0.001, P=0.003), but increased in the PCOS+ IF+ 3-MA group compared to the PCOS+ IF group( P<0.001, P=0.020). Western blotting analysis showed that phosphorylation levels of PI3K, Akt, mTOR, and p70S6K were significantly decreased in the PCOS+ IF group compared with the PCOS group( P=0.002, P=0.001, P=0.001, and P<0.001, respectively), and increased in the PCOS+ IF+ 3-MA group compared with the PCOS+ IF group( P=0.021, P=0.041, P=0.047, and P=0.024, respectively). Conclusions:Intermittent fasting alleviates insulin resistance in a PCOS mouse model through inhibitiing PI3K/Akt/mTOR signaling pathway and promoting autophagy.

Objective:To investigate the association of serum fibroblast growth factor 23(FGF23) level with insulin resistance(IR) and sex hormone levels in patients with polycystic ovary syndrome(PCOS).Methods:A retrospective study was performed in eighty-seven patients with PCOS, fifty-seven patients with simple IR, and sixty-one healthy women who were admitted to Affiliated Hospital of Zunyi Medical University during October 2021 and November 2022. According to the homeostasis model assessment-IR index, all subjects were divided into normal control group( n=61), IR group( n=57), PCOS without IR group(PCOS group, n=15), and PCOS+ IR group( n=72). The levels of serum FGF23, adiponectin, and sex hormones in all groups were compared, and their correlations with glucose and lipid metabolism indicators were analyzed. Results:The FGF23 level was significantly elevated in the IR group, while markedly reduced in the PCOS group and PCOS+ IR group, with the PCOS group showing a significantly lower concentration. The adiponectin levels were significantly decreased in the IR group, PCOS group, and PCOS+ IR group(all P<0.05). The correlation analysis showed that FGF23 level was positively correlated with adiponectin and sex hormone binding globulin, and negatively correlated with luteinizing hormone, luteinizing hormone/follicle stimulating hormone, and free testosterone index(all P<0.05). Logistic regression results indicated that both FGF23 and adiponectin could be used as good indicators for the diagnosis of PCOS and PCOS with IR(all P<0.05). Conclusion:FGF23 is closely related to IR and androgen as well, and under certain conditions, it can reflect the severity of IR and hyperandrogenemia in PCOS patients. The cutoff value of FGF23 obtained in this study can provide a good reference for the diagnosis of PCOS diseases.

Angelicae Sinensis Radix (Danggui) and Ligusticum Chuanxiong Rhizoma (Chuan Xiong) herb-pair (DC) have been frequently used in Traditional Chinese medicine (TCM) prescriptions for hundreds of years to prevent vascular diseases and alleviate pain. However, the mechanism of DC herb-pair in the prevention of liver fibrosis development was still unclear. In the present study, the effects and mechanisms of DC herb-pair on liver fibrosis were examined using network pharmacology and mouse fibrotic model. Based on the network pharmacological analysis of 13 bioactive ingredients found in DC, a total of 46 targets and 71 pathways related to anti-fibrosis effects were obtained, which was associated with mitogen-activated protein kinase (MAPK) signal pathway, hepatic inflammation and fibrotic response. Furthermore, this hypothesis was verified using carbon tetrachloride (CCl

Objective To observe the effects of Huanglian ointment on wound healing of mice with full-thickness skin defect,and to explore the related mechanism.Methods Thirty male C57BL/6J mice were divided into Huanglian ointment group and vehicle group according to the random number table after round wounds of full-thickness skin defect with diameter of 7.5 mm were inflicted on the back of each mouse,with 15 mice in each group.Wounds of mice in Huanglian ointment group and vehicle group were treated with Huanglian ointment and vehicle respectively from post injury day (PID) 1 on,2 times each day.Five mice from each group were selected to observe wound changes on PID 0,3,7,10,and 14,and wound healing rates were calculated.Five mice out of the 10 mice that hadn't been used for general observation in each group were sacrificed on PID 3 and 7 respectively,and 5 mice after being used for general observation in each group were sacrificed on PID 14.Wound and skin tissue within 2 mm from the edge of wound was collected.Histologic scoring was conducted based on the histomorphological observation with HE staining.The expression of double positive cells of alpha smooth muscle actin (αt-SMA) and Ki-67 (myofibroblast) in tissue of wounds of mice was observed by immunofluorescence staining.Protein expressions of transforming growth factor beta (TGF-β) and collagen in tissue of wounds of mice were determined by enzyme-linked immunosorbent assay.Data were processed with analysis of variance for repeated measurement,analysis of variance of factorial design,t test of two independent samples,one-way analysis of variance,and Bonferronni test or correction.Results (1) Wounds of mice in two groups were red and swollen on PID 0,while they were neither red nor swollen with scabs on PID 3 and 7.On PID 10,woundsof mice in Huanglian ointment group contracted obviously,while the contracted wounds of mice in vehicle group were smaller than those in Huanglian ointment group.On PID 14,wounds of most mice in Huanglian ointment group were healed,while wounds of some mice in vehicle group failed to heal.Wound healing rates of mice in two groups were close on PID 3 and 7 (with t values respectively 0.64 and 1.90,P values above 0.05).Wound healing rates of mice in Huanglian ointment group on PID 10 and 14 were (76 ±7)％ and (93 ±5)％ respectively,significantly higher than those of vehicle group [(48 ± 9) ％ and (68 ± 11) ％,with t values respectively 7.44 and 3.89,P values below 0.01].Wound healing rates of mice in two groups on PID 7,10,and 14 were significantly higher than those on the previous time points of the same group (with P values below 0.01).(2) Histologic scores of wounds of mice in two groups were close on PID 3 (t =-0.76,P ＞0.05).Histologic scores of wounds of mice in Huanglian ointment group on PID 7 and 14 were (7.0 ± 1.6) and (11.6 ± 2.1) points respectively,significantly higher than those of vehicle group [(4.2 ± 1.3) and (7.2 ± 1.3) points,with t values respectively 1.96 and 2.50,P ＜ 0.05 or P ＜ 0.01].Histologic scores of wounds of mice in two groups on PID 7 and 14 were significantly higher than those on the previous time points of the same group (with P values below 0.01).(3) Percentages of double positive cells of α-SMA and Ki-67 in tissue of wounds of mice in Huanglian ointment group on PID 3 and 7 were (35:± 12)％ and (62 ± 10) ％ respectively,significantly higher than those of vehicle group [(17 ± 12) ％ and (34 ± 6) ％,with t values respectively-2.48 and-5.25,P ＜0.05 or P ＜0.01].The percentage of double positive cells of α-SMA and Ki-67 in tissue of wounds of mice in Huanglian ointment group on PID 14 was (25 ± 5) ％,significantly lower than that of vehicle group [(44 ± 17) ％,t =2.50,P ＜ 0.05].The percentage of double positive cells of α-SMA and Ki-67 in tissue of wounds of mice on PID 7 was significantly higher than that on PID 3 or 14 in Huanglian ointment group (with P values below 0.01).Percentages of double positive cells of α-SMA and Ki-67 in tissue of wounds of mice on PID 7 and 14 were significantly higher than those on the previous time points in vehicle group (with P values below 0.05).(4) Protein expressions of TGF-β in tissue of wounds of mice in Huanglian ointment group on PID 3 and 7 were (396 ± 45) and (722 ± 96) pg/mL respectively,significantly higher than those of vehicle group [(290 ± 42) and (382 ± 62) pg/mL,with t values respectively-8.17 and-6.65,P values below 0.01].Protein expressions of TGF-β in tissue of wounds of mice in two groups were close on PID 14 (t =1.60,P ＞ 0.05).The protein expression of TGF-β in tissue of wounds of mice in Huanglian ointment group on PID 7 was significantly higher than that on P1D 3 or 14 (with P values below 0.01).Protein expressions of TGF-β in tissue of wounds of mice in vehicle group on PID 7 and 14 were significantly higher than those on the previous time points (with P values below 0.05).Protein expressions of collagen in tissue of wounds of mice in two groups were close on PID 3 (t =1.99,P ＞ 0.05).Protein expressions of collagen in tissue of wounds of mice in Huanglian ointment on PID 7 and 14 were (47 ± 10) and (70 ± 14) ng/mL respectively,significantly higher than those of vehicle group [(34 ± 10) and (42 ± 12) ng/mL,with t values respectively 3.15 and 3.52,P ＜ 0.05 or P ＜ 0.01].Protein expressions of collagen in tissue of wounds of mice in two groups on PID 7 and 14 were significantly higher than those on the previous time points of the same group (P ＜ 0.05 or P ＜ 0.01).Conclusions Huanglian ointment can promote wound healing of full-thickness skin defect of mice through increasing production of myofibroblasts and protein expressions of TGF-β and collagen.

Bioassay, which construct the characteristics consistents with Chinese medical science, is the core mode and methods for the quality control of Chinese materia medica. Taking the bioassay of Radix Isatidis as an example, the contribution, status and application of bioassay in the quality control of Chinese materia medica were introduced in this article, and two key issue (the selection of reference and measurement methods) in the process of establishing bioassay were also explained. This article expects to provide a reference for the development and improvement of the bioassay of Chinese materia medica in a practical manipulation level.
Aconitum ; Materia Medica ; Medicine, Chinese Traditional ; utilization ; Quality Control