OBJECTIVE To study the anti-fatigue effects of differently-cultivated Astragalus extract in a hypoxic environment of the plateau and explore the related mechanisms.METHODS Fifty-six male KM mice were randomly divided into the hypoxic swimming control(HSC)group,imitation wild Astragalus extract(IWA)430,860 and 1 720 mg·kg-1 groups,and cultivated Astragalus extract(CA)463,925 and 1 850 mg·kg-1 groups.The drug was administered by gavage once daily for 15 days,while body mass was monitored every three days.After 15 days of gavage,the mice were subjected to load swimming(5%body weight)in a hypobaric chamber(simulating a 4 000 m altitude),with exhaus-tive swimming time measured to identify the optimal dosage.Following randomization,fifty male KM mice were assigned to five groups:normoxic control(NC),hypoxic control(HC),HSC,IWA 860 and CA 925 mg·kg-1.All groups underwent daily gavage for 15 d before 90 min non-weight-bearing swimming was conducted in the HSC,IWA 860 and CA 925 mg·kg-1 groups within a hypobaric chamber,followed by immediate measurement of muscle strength.Hematoxylin-eosin(HE)staining was used to observe the histopathological changes in liver and gastrocnemius muscle tissues.Blood urea nitrogen(BUN),blood glucose(BG)and serum lactic acid(LA),glutathione(GSH),glutathione peroxidase(GSH-Px),malondialdehyde(MDA),superoxide dismutase(SOD),liver glycogen(LG)and muscle glycogen(MG)in livers and muscles,and total antioxidant capacity(T-AOC)and reactive oxygen species(ROS)in muscles were measured by commercial kits.Taurine and hypotaurine were measured by HPLC.Enzyme-linked immunosorbent assay(ELISA)was used for cysteine sulfenic acid decarboxylase(CSAD)measure-ment.Western blotting was used to detect protein expressions of phosphatidylinositol 3-kinase(PI3K),protein kinase B(Akt),nuclear factor E2-related factor 2(Nrf2),and heme oxygenase-1(HO-1)in skeletal muscles.RESULTS Compared with the HSC group,the swimming time was prolonged in IWA 463,IWA 860,CA 925 and CA 1 850 mg·kg-1 groups.Compared with the HSC group,the muscle strength of mice in the IWA 860 mg·kg-1 group and the CA 925 mg·kg-1 group was significantly increased,histo-pathological damage in the liver and gastrocnemius muscle was reduced,serum levels of LA and BUN were significantly decreased,levels of BG,LG and MG were significantly increased,levels of GSH,GSH-Px and SOD were significantly increased,contents of MDA were significantly decreased,expressions of CSAD were significantly increased in liver tissue,contents of GSH,T-AOC,taurine and hypotaurine were significantly increased,levels of ROS were significantly decreased,and protein expressions of PI3K,Akt,Nrf2,HO-1 were significantly upregulated in muscle tissues.CONCLUSION Under simulated high-altitude hypoxic conditions,extracts of Astragalus membranaceus cultivated by two methods consis-tently exhibit anti-fatigue effects.Its mechanisms may be mitigating oxidative stress,augmenting taurine and hypotaurine metabolic regulation,and activating PI3K/Akt and Nrf2/HO-1 signaling pathways.IWA has a better anti-fatigue effect than CA.

OBJECTIVE To study the anti-fatigue effects of differently-cultivated Astragalus extract in a hypoxic environment of the plateau and explore the related mechanisms.METHODS Fifty-six male KM mice were randomly divided into the hypoxic swimming control(HSC)group,imitation wild Astragalus extract(IWA)430,860 and 1 720 mg·kg-1 groups,and cultivated Astragalus extract(CA)463,925 and 1 850 mg·kg-1 groups.The drug was administered by gavage once daily for 15 days,while body mass was monitored every three days.After 15 days of gavage,the mice were subjected to load swimming(5%body weight)in a hypobaric chamber(simulating a 4 000 m altitude),with exhaus-tive swimming time measured to identify the optimal dosage.Following randomization,fifty male KM mice were assigned to five groups:normoxic control(NC),hypoxic control(HC),HSC,IWA 860 and CA 925 mg·kg-1.All groups underwent daily gavage for 15 d before 90 min non-weight-bearing swimming was conducted in the HSC,IWA 860 and CA 925 mg·kg-1 groups within a hypobaric chamber,followed by immediate measurement of muscle strength.Hematoxylin-eosin(HE)staining was used to observe the histopathological changes in liver and gastrocnemius muscle tissues.Blood urea nitrogen(BUN),blood glucose(BG)and serum lactic acid(LA),glutathione(GSH),glutathione peroxidase(GSH-Px),malondialdehyde(MDA),superoxide dismutase(SOD),liver glycogen(LG)and muscle glycogen(MG)in livers and muscles,and total antioxidant capacity(T-AOC)and reactive oxygen species(ROS)in muscles were measured by commercial kits.Taurine and hypotaurine were measured by HPLC.Enzyme-linked immunosorbent assay(ELISA)was used for cysteine sulfenic acid decarboxylase(CSAD)measure-ment.Western blotting was used to detect protein expressions of phosphatidylinositol 3-kinase(PI3K),protein kinase B(Akt),nuclear factor E2-related factor 2(Nrf2),and heme oxygenase-1(HO-1)in skeletal muscles.RESULTS Compared with the HSC group,the swimming time was prolonged in IWA 463,IWA 860,CA 925 and CA 1 850 mg·kg-1 groups.Compared with the HSC group,the muscle strength of mice in the IWA 860 mg·kg-1 group and the CA 925 mg·kg-1 group was significantly increased,histo-pathological damage in the liver and gastrocnemius muscle was reduced,serum levels of LA and BUN were significantly decreased,levels of BG,LG and MG were significantly increased,levels of GSH,GSH-Px and SOD were significantly increased,contents of MDA were significantly decreased,expressions of CSAD were significantly increased in liver tissue,contents of GSH,T-AOC,taurine and hypotaurine were significantly increased,levels of ROS were significantly decreased,and protein expressions of PI3K,Akt,Nrf2,HO-1 were significantly upregulated in muscle tissues.CONCLUSION Under simulated high-altitude hypoxic conditions,extracts of Astragalus membranaceus cultivated by two methods consis-tently exhibit anti-fatigue effects.Its mechanisms may be mitigating oxidative stress,augmenting taurine and hypotaurine metabolic regulation,and activating PI3K/Akt and Nrf2/HO-1 signaling pathways.IWA has a better anti-fatigue effect than CA.

Objective:To preliminarily investigate the effect of blue light on the biological activity of human skin keratinocytes, fibroblasts and melanocytes.Methods:Discarded foreskin tissues were collected from 10 healthy children aged from 3 to 12 years after circumcision surgery in the First Affiliated Hospital of Kunming Medical University from June 2021 to December 2021. After epidermis-dermis separation, selective culture was performed to isolate keratinocytes, fibroblasts, and melanocytes. According to the pre-experiment results, the above three types of cells were irradiated with 440 - 450 nm blue light at doses of 0, 5, 10, 20, 30, and 40 J/cm 2, and then continued to be cultured for 0, 6, 24, and 48 hours. Cell counting kit 8 (CCK8) assay was performed to evaluate cellular proliferative activity at each time point, enzyme-linked immunosorbent assay (ELISA) to detect levels of interleukin (IL) -18, IL-33, nerve growth factor (NGF), and granulocyte-macrophage colony-stimulating factor (GM-CSF) secreted by keratinocytes, as well as levels of IL-33 and keratinocyte growth factor (KGF) secreted by fibroblasts, NaOH lysis method to determine melanin synthesis rates in melanocytes, and Western blot analysis to determine the relative expression of tyrosinase (TYR), tyrosine-related protease 1 (TRP-1) and dopachrome isomerase (DCT) in melanocytes. Two-way analysis of variance was used to analyze group effects, time effects and interaction effects. Results:After irradiation with blue light, the cellular proliferative activity significantly differed among different doses of blue light irradiation groups and different time points in keratinocytes ( Ftime = 516.20, Fdose = 421.20, Finteraction = 25.05, all P < 0.003), fibroblasts ( Ftime = 129.30, Fdose = 477.80, Finteraction = 10.91, all P < 0.003), and melanocytes ( Ftime = 77.61, Fdose = 138.70, Finteraction = 3.50, all P < 0.003) ; immediately after irradiation, the proliferative activity of keratinocytes and fibroblasts was significantly lower in the 20 - 40 J/cm 2 blue light group than in the 0 J/cm 2 blue light group (all P < 0.003), and the proliferative activity of melanocytes was significantly higher in the 5 J/cm 2 blue light group than in the 0 J/cm 2 blue light group ( P < 0.003) ; the proliferative activity of the 3 types of cells showed decreasing trends with the increase of blue light irradiation doses and culture time. ELISA showed that the concentrations of IL-18, IL-33, NGF, and GM-CSF secreted by keratinocytes, as well as the concentrations of IL-33 and KGF secreted by fibroblasts, tended to increase with the increase of blue light irradiation doses and culture time. The melanin synthesis rates in melanocytes significantly differed among different doses of blue light irradiation groups and different time points ( Ftime = 833.50, Fdose = 249.40, Finteraction = 81.38, all P < 0.003) ; during 0 - 24 hours after blue light irradiation, the melanin synthesis rates tended to increase with the increase of blue light irradiation doses and time; during 24 - 48 hours, the melanin synthesis rates showed decreasing trends with the increase of blue light irradiation doses and culture time compared with that at 24 hours after irradiation; 24 hours after irradiation, the melanin synthesis rates were significantly higher in the 5, 10, 20, 30 and 40 J/cm 2 blue light groups (159.50% ± 10.88%, 218.76% ± 8.49%, 333.72% ± 7.72%, 393.29% ± 6.00%, 427.21% ± 8.39%, respectively) than in the 0 J/cm 2 blue light group (102.29% ± 6.57%, all P < 0.003). The relative expression of TYR ( Ftime = 67.94, Fdose = 28.99, Finteraction = 3.71, all P < 0.003), TRP-1 ( Ftime = 21.73, Fdose = 8.38, both P < 0.003) and DCT ( Ftime = 34.51, Fdose = 11.79, both P < 0.003) in melanocytes significantly differed among different doses of blue light irradiation groups and different time points, and tended to increase with the increase of blue light irradiation doses and culture time. Conclusion:Blue light irradiation at doses of 5 - 40 J/cm 2 could inhibit the proliferative activity of human skin keratinocytes, fibroblasts, and melanocytes, and the inhibitory effect tended to increase with the increase of blue light irradiation doses, except an enhancing effect on the proliferative activity of melanocytes observed immediately after irradiation with blue light at 5 J/cm 2; additionally, blue light irradiation at 5 - 40 J/cm 2 could enhance the expression of melanin synthesis-related enzymes in melanocytes, and increase the melanin synthesis rate in melanocytes over a short period of time.

OBJECTIVE To establish the fingerprints of c ultivated and wild Anemarrhena asphodeloides，and to identify their differential components. METHODS Using an evaporative light-scattering detector ， the high performance liquid chromatography combined with Similarity Evaluation System of TCM Chromatographic Fingerprint （2012 edition） were used to establish fingerprints of 14 batches of cultivated A. asphodeloides and 14 batches of wild medicinal materials ，and evaluate their similarity. The common peaks were identified by comparison with the chromatogram of the mixed control. At the same time ，the contents of components corresponding to common peaks in cultivated and wild A. asphodeloides were determined. The principal component analysis and orthogonal partial least squares discrimination analysis were adopted to identify differential components of them ，and compare the contents of them. RESULTS Among 28 batches of A. asphodeloides ，10 common peaks were found ，i.e. neomangiferin（peak 1），mangiferin（peak 2），isomangiferin（peak 3），timosaponin B Ⅱ（peak 7），timosaponin B Ⅲ（peak 8）， timosaponin Ⅰ（peak 9），timosaponin A Ⅲ（peak 10）. The similarities of fingerprints of samples with control fingerprint were no less than 0.963. The average total contents of seven components in cultivated and wild A. asphodeloides were 74.18 and 84.72 mg/g， respectively；there was statistical significance （P＜0.05）. The cultivated and wild A. asphodeloides could be divided into two categories. The differential components were neomangiferin ，mangiferin，timosaponin B Ⅱ and timosaponin A Ⅲ（VIP values were all higher than 1）. The content of neomangiferin in cultivated products was significantly higher than that in wild products （P＜ 0.05），and the contents of mangiferin ，timosaponin B Ⅱ and ti mosaponin A Ⅲ were significantly lower than those in wild products （P＜0.05）. CONCLUSIONS Fingerprint of A. asphodeloides is established ，and differential components of cultivated and wild A. asphodeloides are identified primarily.