The 95% ethanol extract from bulbs of Narcissus tazetta var. chinensis(BNTC) was eluted with 30%, 60%, and pure methanol on D-101 macroporous resin. The elution fractions were isolated and purified by silica gel column chromatography, thin layer chromatography, D-101 macroporous resin, semi-preparative high performance liquid chromatography(HPLC), and HPLC. The purified compounds were identified using one-dimensional and two-dimensional spectroscopy, high-resolution mass spectrometry, and other techniques. A total of 15 compounds were isolated and identified as 5-(4-hydroxy-3-methoxyphenyl)-3-(4-hydroxyphenyl)-N-methyl-3,6-dihydropyridine-2(1H)-one(1), 3,5-di(hydroxyphenyl)-N-methyl-3,6-dihydropyridine-2(1H)-one(2), protocatechualdehyde(3), protocatechuic acid(4), 3,4-dihydroxyacetophenone(5), syringic acid(6), vanillic acid(7), p-hydroxybenzoic acid(8),(2S)-4'-hydroxy-7-methoxyflavan(9), 2,4,6-trimethoxyacetophenone(10), N-trans-ferulic acid p-hydroxyphenylethylamine(11), N-cis-p-coumaroyltyramine(12), N-trans-p-coumaroyltyramine(13), piscidic acid(14), 5-hydroxymethylfurfural(15). Compounds 1 and 2 are new compounds with similar structure that have not been reported yet, named narcissus A and narcissus B. Compounds 8-13 were isolated and identified from the genus Narcissus for the first time, and compounds 14 and 15 were isolated from BNTC for the first time. Compounds 1 and 2 inhibited the release of NO from RAW264.7 cells induced by lipopolysaccharide(LPS)(P<0.001), with compound 1 having an IC_(50) value of(72.76±2.97) μmol·L~(-1) and compound 2 having an IC_(50) value of(63.59±0.96) μmol·L~(-1).
Mice ; Animals ; Narcissus/chemistry* ; Drugs, Chinese Herbal/isolation & purification* ; Plant Roots/chemistry* ; Chromatography, High Pressure Liquid ; Macrophages/immunology* ; RAW 264.7 Cells

OBJECTIVE: To investigate the effects and mechanisms of hydroxysafflor yellow A (HSYA) on replicative senescence in human umbilical cord mesenchymal stem cells (hUC-MSCs). METHODS: hUC-MSCs were cultured to construct a replicative senescence model through continuous amplification in vitro. Cells at passage 2 served as the control group, while cells at passage 10 were designated as the senescence group. The senescent cells were cultured in a culture medium containing HSYA. Cell viability was detected by the CCK-8 assay, and cell confluence was analyzed using the Incucyte S3 live-cell analysis system. The optimal concentration and time point were determined and utilized for subsequent experiments. Senescent cells were pretreated with 0.01 mg/ml HSYA, and the proportion of senescence-associated β-galactosidase (SA-β-gal) positive cells was detected to assess the senescence state. The relative telomere length was detected by qPCR. Reactive oxygen species (ROS) levels were measured using the fluorescent probe DCFH-DA. Mitochondrial membrane potential was assessed by JC-1 staining. The expression of p53, p16, p21, OCT4, and SOX2 genes was detected by qPCR. The expression of p16, p53, OCT4, and SOX2 proteins was analyzed by Western blot. RESULTS: HSYA significantly decreased the SA-β-gal positive staining rate, inhibited telomere attrition, reduced the ROS accumulation, increased mitochondrial membrane potential in senescent cells. Additionally, HSYA downregulated the expression of p53 and p16, and upregulated the expression of OCT4. HSYA decreased p16 protein level and increased OCT4 and SOX2 protein levels. CONCLUSION HSYA may ameliorate replicative senescence in hUC-MSCs by modulating the p53 and p16 signaling pathways and suppressing oxidative stress.
Humans ; Mesenchymal Stem Cells/drug effects* ; Cellular Senescence/drug effects* ; Chalcone/pharmacology* ; Oxidative Stress/drug effects* ; Quinones/pharmacology* ; Umbilical Cord/cytology* ; Reactive Oxygen Species/metabolism* ; Cells, Cultured ; Cyclin-Dependent Kinase Inhibitor p16/metabolism* ; Tumor Suppressor Protein p53/metabolism* ; Membrane Potential, Mitochondrial ; Cell Proliferation

Chronic cerebral hypoperfusion leads to white matter injury (WMI), which plays a significant role in contributing to vascular cognitive impairment. While 13-docosenamide is a type of fatty acid amide, it remains unclear whether it has therapeutic effects on chronic cerebral hypoperfusion. In this study, we conducted bilateral common carotid artery stenosis (BCAS) surgery to simulate chronic cerebral hypoperfusion-induced WMI and cognitive impairment. Our findings showed that 13-docosenamide alleviates WMI and cognitive impairment in BCAS mice. Mechanistically, 13-docosenamide specifically binds to cannabinoid receptor 1 (CNR1) in oligodendrocyte precursor cells (OPCs). This interaction results in an upregulation of ubiquitin-specific peptidase 33 (USP33)-mediated CNR1 deubiquitination, subsequently increasing CNR1 protein expression, activating the phosphorylation of the AKT/mTOR pathway, and promoting the differentiation of OPCs. In conclusion, our study suggests that 13-docosenamide can ameliorate chronic cerebral hypoperfusion-induced WMI and cognitive impairment by enhancing OPC differentiation and could serve as a potential therapeutic drug.
Animals ; Oligodendrocyte Precursor Cells/metabolism* ; Mice ; Cell Differentiation/drug effects* ; Male ; Receptor, Cannabinoid, CB1/metabolism* ; Mice, Inbred C57BL ; Ubiquitin Thiolesterase/metabolism* ; Ubiquitination/drug effects* ; Carotid Stenosis/complications* ; Cognitive Dysfunction/drug therapy*

Objective To observe the clinical efficacy of acupuncture combined with acupoint catgut embedding therapy on back-shu and front-mu points and external application on shenque(RN8)point for premature ovarian failure(POF).Methods A total of 62 patients with POF were randomly divided into the observation group and the control group,with 31 patients in each group.The observation group was treated with acupuncture combined with acupoint catgut embedding therapy on back-shu and front-mu points and external application on shenque point,and the control group was treated with hormone replacement therapy.After three months of treatment,the clinical efficacy of the two groups was evaluated,and the changes in the traditional Chinese medicine(TCM)syndrome scores,as well as the ovarian volume,number of antral follicle,and antral follicle diameter of the patients in the two groups before and after treatment were observed.The changes of serum follicle stimulating hormone(FSH),luteinizing hormone(LH)and estradiol(E2)levels before and after treatment were compared between the two groups.Results(1)The total effective rate was 93.55%(29/31)in the observation group and 80.64%(25/31)in the control group.The efficacy of the observation group was significantly superior to that of the control group,and the difference was statistically significant(P＜0.05).(2)After treatment,the serum FSH,LH and E2 levels of patients in the two groups were significantly improved(P＜0.05),and the improvement in the observation group was significantly superior to that in the control group,with statistically significant differences(P＜0.05).(3)After treatment,ovarian volume,number of antral follicle,and antral follicle diameter were significantly improved in the two groups(P＜0.05),and the improvement in the observation group was significantly superior to that in the control group,and the difference was statistically significant(P＜0.05).Conclusion The treatment of POF with acupuncture combined with acupoint catgut embedding therapy on back-shu and front-mu points and external application on shenque point can significantly improve the clinical symptoms of the patients,conducive to the recovery of ovarian function,and significantly improve the sex hormone levels of the patients,with precise clinical efficacy.

Objective To explore the potential category characteristics of self-management behavior in patients with lung cancer based on potential profile analysis,and to analyze the characteristic differences and influencing factors of self-management behavior in patients with different categories of lung cancer.Methods A total of 260 patients with lung cancer who had completed the main treatment program and were about to enter the follow-up period in a tertiary A general hospital in Zhejiang Province from July 2022 to May 2023 were selected by convenience sampling method as the investigation subjects.General Information Questionnaire,Lung Cancer Survivor Self-management Behavior Assessment Scale,Strategies Used by People to Promote Health,Hospital Anxiety and Depression Scale and Social Support Rating Scale were used for investigation.Potential profile analysis was used to explore the potential categories of self-management behavior in lung cancer patients and analyze its influencing factors.Results 252 patients were finally included.The results of potential profile analysis showed that lung cancer patients'self-management behavior could be divided into 3 potential categories,namely low self-management behavior-low emotion management group(11.90％),medium self-management behavior-low resource management group(45.24％),and high self-management behavior-low hope management group(42.86％).Logistic regression analysis showed that education level,previous surgery,tumor stage,anxiety level,depression level,self-efficacy level and social support level were the influencing factors of lung cancer patients'self-management behavior(P＜0.05).Conclusion The self-management behavior of lung cancer patients is at a moderate level,and there are obvious classification characteristics.It is suggested that medical staff should carry out personalized intervention measures according to the characteristics of self-management behavior of patients of various categories,so as to improve the level of self-management behavior of lung cancer survivors.

Aim To investigate how the long non-cod-ing RNA uc.48+(lncRNA uc.48+)affected calci-tonin gene-related peptide(CGRP)in the trigeminal ganglion(TG)of rats with trigeminal neuralgia(TN)and its potential mechanism.Methods Chronic con-striction injury of the infraorbital nerve(CCI-ION)in rats was used to create the animal model for trigeminal neuralgia.After modeling,uc.48+siRNA was injec-ted locally via the infraorbital foramen to knock down lncRNA uc.48+,and uc.48+plasmid was transfect-ed into normal rats to over-express lncRNA uc.48+.The face mechanical pain threshold(MWT)of each group was measured by behavioral test,and the content and changes of CGRP in rat TG were observed using qPCR and protein blotting.The change in serum in-flammatory cytokine 1L-1β was determined using ELISA.Results The MWT in TN rats treated with the uc.48+siRNA increased significantly,but the protein and mRNA levels of CGRP in TG decreased significantly(P＜0.01),and the level of 1L-1β de-creased as well(P＜0.01).In addition,the MWT of normal rats transfected with uc.48+plasmid was sig-nificantly diminished,and the mRNA and protein lev-els of CGRP in TG were markedly elevated(P＜0.01),as were the levels of 1L-1β(P＜0.01),compared to normal rats.Conclusions Knocking out uc.48+in TN rats reduces pain,while overexpressing uc.48+exacerbates pain transmission in trigeminal neuralgia.The mechanism by which uc.48+small in-terference inhibits trigeminal neural pathology pain may be through decreasing CGRP expression in TG of rats with TN,therefore ameliorating mechanical pain sensi-tivity.

ObjectiveTo explore the effect of Anmeidan （AMD） on the learning and memory ability of sleep-deprived rats and the mechanism. MethodA total of 50 SD rats were randomized into control group， model group， low-dose AMD group （4.55 g·kg^-1·d^-1）， high-dose AMD group （18.18 g·kg^-1·d^-1）， and estazolam group （0.09 mg·kg^-1·d^-1）. Insomnia was induced in rats with the self-made sleep deprivation box （21 days）. The learning and memory ability of rats was measured by Morris water maze. Immunofluorescence method was employed to detect the number of cells expressing N-myc downstream-regulated gene 2 （NDRG2） and glial fibrillary acidic protein （GFAP） in hippocampus of rats， real-time fluorescent quantitative polymerase chain reaction （Real-time PCR） to determine the mRNA expression of hippocampal NDRG2， glial glutamate transporter-1 （GLT-1）， and GluNR2A and GluNR2B N-methyl-D-aspartate （NMDA） receptor subunits， and Western blot to examine the protein expression of NDRG2 and GLT-1 in hippocampus. ResultCompared with control group， the model group showed increase in the latency to reach the platform and total swimming distance， significant decrease in the total distance moved in the target quadrant， time in target quadrant， and times of crossing the platform （P<0.01）， rise in the number of cells expressing NDRG2 and GFAP in the hippocampal CA1 region （P<0.01） and the mRNA level of NDRG2 and GluNR2B， reduction in the mRNA level of GLT-1 and GluNR2A， elevation in NDRG2 protein expression （P<0.01）， and decrease in GLT-1 protein expression （P<0.01）. In contrast to the model group， low-dose and high-dose AMD improved the learning and memory levels of sleep-deprived rats （P<0.01）， reduced the number of cells expressing NDRG2 and GFAP （P<0.01）， significantly decreased the mRNA expression of NDRG2 and GluNR2B， increased the mRNA expression of GLT-1 and GluNR2A， reduced NDRG2 protein level （P<0.05， P<0.01）， and raised GLT-1 protein level （P<0.01）. ConclusionAMD can improve the learning and memory ability of sleep-deprived rats. The mechanism is the likelihood that it regulates astrocyte activity， thereby affecting the neurotransmitter level and synaptic plasticity in the brain.

Objective:To measure the anatomical parameters of the simulated low tibial tunnel of posterior cruciate ligament (PCL) based on knee CT images so as to provide clinical reference for accurate location of the tunnel.Methods:The CT images of 201 healthy knee joints collected at Department of Orthopedics, The Second Hospital of Lanzhou University from June 2016 to September 2021 were used for simulation of the PCL low tibial tunnel. The anatomical parameters of the tibial tunnel were measured using the RadiAnt DICOM Viewer. The primary measures included the angle between tibial plateau and tibial tunnel (ATPT) and the perpendicular distances from the tibial tunnel entrance and exit point to the tibial plateau (L1 and L2). The secondary measures included the angle between tibial plateau and posterior slope (PSA), the angle between tibial anatomical axis and central line of tibial tunnel (ATAA), the angle between posterior tibial slope line and the central line of tibial tunnel (APST), the anterior and posterior diameter of tibial plateau (APD), the length of posterior tibial slope (LPTS), and the length of tibial tunnel (LTT). The measurement results were analyzed according to the body height (divided into 3 groups: a 1.00 to 1.60 m group, a 1.61 to 1.70 m group, and a ≥1.71 m group) and gender using the software IBM SPSS 26.Results:The primary measures: ATPT was 37.0°±4.5°, and L1 and L2 were respectively (57.8±7.4) mm and (34.5±3.3) mm. The secondary measures: PSA 128.1°±5.4°, ATAA 52.7°±4.1°, APST 89.1°±5.9°, APD was (32.9±2.6) mm, LPTS (20.5±2.4) mm, and LTT (40.9±5.7) mm. After grouping by gender, there was no significant difference in PSA between men and women ( P>0.05) while there were significant differences in the other indexes between men and women ( P<0.05). After grouping by body height, there was no significant difference in ATPT, PSA, APST or ATAA between the 3 groups (1.00 to 1.60 m group, 1.61 to 1.70 m group and ≥1.71 m group) ( P>0.05) while there were significant differences in L1, L2, APD, LPTS and LTT between the 3 groups ( P<0.05). Conclusions:Based on the knee CT images, the primary measures of PCL low tibial tunnel are as follows: the angle between tibial plateau and tibial tunnel is 37.0°±4.5°, and the perpendicular distances from the tibial tunnel entrance and exit point to the tibial plateau are (57.8±7.4) mm and (34.5±3.3) mm, respectively. Gender and body height are the important factors influencing the above measurement outcomes.

Objective::To observe the effect of Bimin decoction(BMD) on nuclear factor-kappa B(NF-κB) signaling pathway and aquaporin 5(AQP5) expression in allergic rhinitis (AR) rats with lung and spleen Qi deficiency syndrome(LSQDS), in order to study the mechanism in treating AR. Method::Fifty-six SD rats were randomly divided into seven groups: control group, AR group, LSQDS AR group, BMD low dose two weeks group and four weeks group, BMD high dose two weeks group and four weeks group. The control group did not intervened, the AR group established the AR disease model with ovalbumin (OVA) as the allergen, the other five groups established the LSDQS model with smoke and senna gavage, and also established the AR disease model with OVA sensitization at the same time as the AR group. After the model was established successfully, four BMD intervention groups were separately given low dose BMD (11.3 g·kg^-1) for 2 weeks and 4 weeks, and high dose BMD (22.6 g·kg^-1) for 2 weeks and 4 weeks. To observe the general situation of the rats, hematoxylin eosin (HE) staining was used to observe the pathological changes of the nasal mucosa, immunohistochemistry and Western blot were used to detect the expression levels of NF-κB and AQP5, real-time fluorescent quantitative polymerase chain reaction technique (Real-time PCR)was used to detect mRNA levels of interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6). Result::The typical AR symptoms were found in AR rats, the AR symptoms and lung and spleen Qi deficiency symptoms were found in AR rats with LSQDS at the same time, and the AR symptoms and lung and spleen Qi deficiency symptoms were significantly improved after the intervention of BMD. Compared with the control group, the typical histopathological changes of nasal mucosa were found in AR group and LSQDS AR group, with a higher behavioral score (P<0.05), and the expression of NF-κB and AQP5 protein increased (P<0.05), the expression of IL-1β, TNF-α, IL-6 and AQP5 mRNA increased (P<0.05). Compared with AR group, the pathological changes of nasal mucosa in LSQDS AR group were more serious, and the expression of NF-κB protein in nucleus increased (P<0.05), the expression of TNF-α and AQP5 mRNA increased (P<0.05). Compared with LSQDS AR group, the pathological changes of nasal mucosa in the groups which interfered by BMD were improved, and the expression of NF-κB and AQP5 protein decreased (P<0.05), the expression of IL-1β, TNF-α, IL-6 and AQP5 mRNA decreased (P<0.05). Compared with BMD low-dose two-week group, the expression of NF-κB protein in nucleus decreased (P<0.05) in BMD high dose four week group. Conclusion::Compared with AR group, the AR condition of the rats with LSQDS is more serious under the same allergen stimulation, BMD can treat AR and reduce the over secretion of glands, which may be related to inhibit the expression of AQP5 by inhibiting the NF-κB signaling pathway.