Objective:To observe the effect of the Yushen Yangluan decoction on ovarian reserve function in mice with diminished ovarian reserve (DOR) and to explore its mechanism of action based on the hypoxia-inducible factor (HIF)-1α/vascular endothelial growth factor A (VEGFA) signaling pathway.Methods:Mechanism analysis was conducted using artificial intelligence-based network pharmacology. Fifty 6-week-old female C57BL/6 mice were randomly divided into five groups according to body weight, with 10 mice in each group. The groups were as follows: control group, model group, low-dose Traditional Chinese Medicine (TCM) group, medium-dose TCM group, and high-dose TCM group. Hematoxylin-eosin staining was used to observe the number of antral follicles in the ovaries, enzyme-linked immunosorbent assay was employed to detect serum hormone levels, and TdT-mediated dUTP nick end labeling (TUNEL) was used to assess apoptosis in ovarian granulosa cells. Immunohistochemistry (IHC) was performed to detect ovarian expression of HIF-1α, and Western blotting was used to measure the protein expressional levels of HIF-1α, VEGFA, and B-cell lymphoma-2 (BCL2) in the ovaries.Results:After treating DOR model mice with the Yushen Yangluan decoction at low, medium, and high dosages, the ovarian antral follicle count of the mice in the TCM groups was all higher than that in the model group ( P=0.024, P<0.001, P<0.001), the follicle-simulating hormone level was lower than that in the model group ( P=0.003, P<0.001, P<0.001), and the serum anti-Müllerian hormone level in the medium-and high-dose TCM groups was higher than that in the model group ( P=0.001, P<0.001). Network pharmacology and molecular docking suggested that the formula may influence the HIF signaling pathway through core targets such as HIF-1α, BCL2 and VEGFA. TUNEL assays indicated that the apoptosis rate of ovarian granulosa cells in the low-, medium-, and high-dose TCM groups was significantly lower than that in the model group ( P=0.017, P=0.003, P<0.001). Immunohistochemistry results showed that the expression level of HIF-1α in the granulosa cells of the follicles in the low-, medium- and high-dose TCM groups was significantly higher than that in the model group ( P=0.008, P<0.001, P<0.001). Western blotting analysis indicated that the protein expression level of HIF-1α in medium-, and high-dose TCM groups was significantly higher than that in the model group ( P=0.004, P<0.001), and the protein expression levels of VEGFA and BCL2 in low-, medium-, and high-dose TCM groups were significantly higher than those in the model group ( P=0.003, P=0.002, P=0.001; P=0.029, P=0.007, P=0.001). Conclusion:The Yushen Yangluan decoction may exert its therapeutic effect by inhibiting apoptosis of ovarian granulosa cells through the HIF-1α/VEGFA signaling pathway.

Objective:To observe the effect of the Yushen Yangluan decoction on ovarian reserve function in mice with diminished ovarian reserve (DOR) and to explore its mechanism of action based on the hypoxia-inducible factor (HIF)-1α/vascular endothelial growth factor A (VEGFA) signaling pathway.Methods:Mechanism analysis was conducted using artificial intelligence-based network pharmacology. Fifty 6-week-old female C57BL/6 mice were randomly divided into five groups according to body weight, with 10 mice in each group. The groups were as follows: control group, model group, low-dose Traditional Chinese Medicine (TCM) group, medium-dose TCM group, and high-dose TCM group. Hematoxylin-eosin staining was used to observe the number of antral follicles in the ovaries, enzyme-linked immunosorbent assay was employed to detect serum hormone levels, and TdT-mediated dUTP nick end labeling (TUNEL) was used to assess apoptosis in ovarian granulosa cells. Immunohistochemistry (IHC) was performed to detect ovarian expression of HIF-1α, and Western blotting was used to measure the protein expressional levels of HIF-1α, VEGFA, and B-cell lymphoma-2 (BCL2) in the ovaries.Results:After treating DOR model mice with the Yushen Yangluan decoction at low, medium, and high dosages, the ovarian antral follicle count of the mice in the TCM groups was all higher than that in the model group ( P=0.024, P<0.001, P<0.001), the follicle-simulating hormone level was lower than that in the model group ( P=0.003, P<0.001, P<0.001), and the serum anti-Müllerian hormone level in the medium-and high-dose TCM groups was higher than that in the model group ( P=0.001, P<0.001). Network pharmacology and molecular docking suggested that the formula may influence the HIF signaling pathway through core targets such as HIF-1α, BCL2 and VEGFA. TUNEL assays indicated that the apoptosis rate of ovarian granulosa cells in the low-, medium-, and high-dose TCM groups was significantly lower than that in the model group ( P=0.017, P=0.003, P<0.001). Immunohistochemistry results showed that the expression level of HIF-1α in the granulosa cells of the follicles in the low-, medium- and high-dose TCM groups was significantly higher than that in the model group ( P=0.008, P<0.001, P<0.001). Western blotting analysis indicated that the protein expression level of HIF-1α in medium-, and high-dose TCM groups was significantly higher than that in the model group ( P=0.004, P<0.001), and the protein expression levels of VEGFA and BCL2 in low-, medium-, and high-dose TCM groups were significantly higher than those in the model group ( P=0.003, P=0.002, P=0.001; P=0.029, P=0.007, P=0.001). Conclusion:The Yushen Yangluan decoction may exert its therapeutic effect by inhibiting apoptosis of ovarian granulosa cells through the HIF-1α/VEGFA signaling pathway.

Objective:To explore the effect of adult growth hormone deficiency on cognitive function in adults.Methods:A total of 19 hypophyseal or craniopharyngioma patients who underwent surgical treatment and were diagnosed with adult growth hormone deficiency in Department of Endocrinolog, the Second Affiliated Hospital, School of Medicine, Zhejiang University from June 2017 to June 2018 were selected as the case group, and 19 normal people were included as the control group. All the members were assessed with the cognitive function scale and brain functional magnetic resonance examination, data between the groups were analyzed.Results:The body weight within a year of case group was significantly increased than that of the control group( P=0.017). Compared with the control group, the case group was relatively inattentive and had decreased memory(Time of stroop color words test-a, test-c, and trail-making test-A, P values were 0.009, 0.018, 0.020 respectively; Auditory word learning test N6, P=0.008). The executive function and language ability of the case group were weakened compared with the control group(Raven′s matrices score E1-E12, P=0.022; Time cost and the number of arrivals in 1 min of connection test B, P values were 0.023, 0.004; Symbol digit modalities test, P=0.037; The number of words spoken in 46-60 s and total number in 0-60 s of the case group was less than the control, P values were 0.030, 0.006). The general mental state of the case group was worse than the control group( P=0.018). The accuracy of the 2-back task of the case group was significantly lower and the activation signal of the left frontal lobe in the case group was significantly weaker( P<0.005). Conclusions:Adult growth hormone deficiency may increase obesity risk and have a detrimental influence on patients′ overall mental health, resulting in varying degrees of cognitive impairment. Working memory impairments associated with adult growth hormone deficiency may be a result of decreased frontal lobe brain activity.

ObjectiveTo investigate the effect of porcine large intestine-processed Dahuang （Radix et Rhizoma Rhei） on defecation in constipation model mice and the possible mechanism. MethodsFifty Kunming mice were randomized to blank group （n=10） and model group （n=40）. Loperamide suspension at the dose of 8 mg/（kg·d） was given by gavage for four consecutive days to establish a model of constipation. The 24 successfully modeled mice were randomly divided into model group， processed Dahuang group， lactulose group， raw Dahuang group， with six mice in each group. Moreover， six randomly selected mice were chosen as control group. Since the fifth day， 8 mg/（kg·d） of loperamide suspension by gavage was given to the model group， processed Dahuang group， raw Dahuang group， and lactulose group； two hours later， the processed and raw Dahuang groups were administered with 0.6 g/（kg·d） of processed and raw Dahuang suspension， respectively， while the lactulose group was given 0.6 g/（kg·d） of latulose suspension， and the blank group and the model group were given 0.2 ml/10 g of distilled water by gavage， all for four days. The general condition， body weight after the last gavage， number of fecal particles within six hours， fecal wet weight， fecal water content ratio， intestinal propulsion rate and colonic histology changes by HE staining of each group were detected. ResultsThe body weight of the mice in the raw Dahuang group was significantly lighter than that in the other groups （P＜0.05 or P＜0.01）. The number of fecal particles， fecal wet weight and intestinal propulsion rate of mice significantly decreased in the model group than in the blank group （P＜0.05 or P＜0.01）. Compared to those in the model group， the number of fecal particles and fecal wet weight in the processed Dahuang group， lactulose group and raw Dahuang group significantly increased， and the fecal water content ratio in the raw Dahuang group increased as well （P＜0.05 or P＜0.01）. Compared to those in the processed Dahuang group， the number of fecal particles and fecal wet weight in the raw Dahuang group decreased， while the fecal water content ratio increased （P＜0.05 or P＜0.01）， and the fecal water content ratio in the lactulose group increased significantly （P＜0.05）. The intestinal propulsion rate in the processed Dahuang group was higher than that in the model group， lactulose group and raw Dahuang group （P＜0.05 or P＜0.01）. Histopathological analysis showed that the colonic crypts and goblet cells in the blank group were normal and clear， and the colonic muscular layer was thicker. The colonic crypts of the mice in the model group were damaged， with reduced goblet cells to varying degrees and changed colonic muscularis. In the lactulose group and raw Dahuang group， part of the crypts were broken， and the goblet cells were damaged to varying degrees， while in the processed Dahuang group， still the colonic tissue structure of the mice was relatively clear， and the colonic crypts and goblet cells were relatively normal， with thickened muscular layer of the colon. ConclusionPorcine large intestine-processed Dahuang could improve defecation in constipation model mice， and reduce the drastic purgation function of raw Dahuang， for which the mechanism may be related to the protection of colon histopathological damage.