1.Effect of Huayu Jiedu Prescription on Oxygen-glucose Deprivation-induced Injury in Brain Microvascular Endothelial Cells Based on PI3K/Akt/mTOR Autophagy Related Pathway
Xun PENG ; Yujia LI ; Dingxiang LI ; Yihui DENG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(9):111-121
ObjectiveTo investigate the effects of Huayu Jiedu prescription on brain microvascular endothelial cells (BMECs) after oxygen-glucose deprivation (OGD) injury and to explore its intervention mechanisms. MethodsThe cell counting kit-8 (CCK-8) assay was used to determine the optimal OGD duration and the effective concentration of Huayu Jiedu prescription-containing serum. Cells were randomly divided into the blank serum medium group (KBXQ), model group (OGD), HYXQ group (OGD + Huayu Jiedu prescription-containing serum), and 3-methyladenine (3-MA) group (OGD + 3-MA). Cell morphology was observed under an inverted microscope. The numbers of autophagosomes and autolysosomes in cells were observed by transmission electron microscopy. Cell viability was determined using the CCK-8 assay. Cell apoptosis rate was detected using the TdT-mediated dUTP nick-end labeling (TUNEL) assay. The expression levels of microtubule-associated protein 1 light chain 3 (LC3) and Occludin were detected by immunofluorescence. The permeability of the cell monolayer was also measured. Cells were further randomly divided into the KBXQ group, model group (OGD), HYXQ group, phosphatidylinositol-3 kinase (PI3K) inhibitor (LY294002) group (OGD + LY294002), and HYXQ + LY294002 group (OGD + Huayu Jiedu prescription-containing serum + LY294002). Western blot analysis was used to detect the expression levels of the autophagy-related key molecule yeast Atg6 homolog 1 (Beclin1), LC3Ⅱ/LC3Ⅰ, selective autophagy adaptor protein (p62), Occludin, phosphorylated (p)-PI3K, PI3K, phosphorylated protein kinase B (p-Akt), Akt, phosphorylated mammalian target of rapamycin (p-mTOR), and mTOR. ResultsOGD for 6 h was selected as the optimal modeling condition, and 5% was determined as the optimal volume fraction of Huayu Jiedu prescription-containing serum. Compared with the KBXQ group, the model group showed obvious cell damage under the inverted microscope, and transmission electron microscopy revealed markedly increased numbers of autophagosomes and autolysosomes. Cell viability was significantly decreased (P<0.01), apoptosis rate was significantly increased (P<0.01), LC3 fluorescence intensity was significantly increased (P<0.01), Occludin fluorescence intensity was significantly decreased (P<0.01), and monolayer permeability was significantly increased (P<0.01). Compared with the model group, cell damage in the HYXQ group and the 3-MA group was significantly improved, the numbers of autophagosomes and autolysosomes were markedly reduced, cell viability was significantly increased (P<0.01), apoptosis rate was significantly decreased (P<0.01), LC3 fluorescence intensity was significantly decreased (P<0.01), Occludin fluorescence intensity was significantly increased (P<0.01), and monolayer permeability was reduced (P<0.05). Western blot results showed that, compared with the KBXQ group, the model group exhibited significantly increased expression of Beclin1 and LC3Ⅱ/LC3Ⅰ (P<0.01), while the expression levels of p62, Occludin, p-PI3K/PI3K, p-Akt/Akt, and p-mTOR/mTOR were significantly decreased (P<0.01). Compared with the model group, the HYXQ group showed significantly decreased expression of Beclin1 and LC3Ⅱ/LC3Ⅰ (P<0.01) and significantly increased expression of p62, Occludin, p-PI3K/PI3K, p-Akt/Akt, and p-mTOR/mTOR (P<0.01). In the LY294002 group, Beclin1 and LC3Ⅱ/LC3Ⅰ expression were significantly increased (P<0.05, P<0.01), whereas the expression levels of p62, Occludin, p-PI3K/PI3K, p-Akt/Akt, and p-mTOR/mTOR were significantly decreased (P<0.01). Compared with the LY294002 group, the HYXQ + LY294002 group showed significantly decreased expression of Beclin1 and LC3Ⅱ/LC3Ⅰ (P<0.01) and significantly increased expression of p62, Occludin, p-PI3K/PI3K, p-Akt/Akt, and p-mTOR/mTOR (P<0.01). ConclusionHuayu Jiedu prescription has a protective effect on BMECs after OGD injury, which may be achieved by activating the PI3K/Akt/mTOR autophagy-related signaling pathway and inhibiting excessive autophagy, thereby protecting Occludin protein expression and endothelial barrier function.
2.Clinical characteristics and changes and significance of TH17/Treg cells and lymphocyte subsets in adult secondary hemoph-agocytic syndrome
Yujia WANG ; Zhe PENG ; Chenlu WANG ; Xin LI ; Tao ZHANG
Chinese Journal of Clinical Laboratory Science 2024;42(9):659-663
Objective To investigate the clinical characteristics of adult secondary hemophagocytic syndrome(sHLH)and analyze the changes and significance of TH17/Treg cells and lymphocyte subsets,providing favorable references for clinical work.Methods 82 a-dult sHLH patients admitted to our hospital from April 2021 to March 2023 were selected.On the second day of admission,we observed their clinical characteristics and detected the levels of TH17/Treg cells(TH17,Treg,TH17/Treg)and lymphocyte subsets(CD3+,CD4+,CD8+,CD19+,CD16+CD56+).The patients were given the HLH-2004 treatment regimen and followed up for 6 months.Ac-cording to the treatment results,they were divided into the survival group(n=57)and fatality group(n=25).The levels of TH17/Treg cells and lymphocyte subsets on the second day of admission between the two groups were compared.The correlation between TH17/Treg cells and lymphocyte subsets and the value of TH17/Treg cells in evaluating the fatality of adult sHLH patients were ana-lyzed.Results All 82 adult sHLH patients had elevated serum beta-2 microglobulin and ferritin levels,and more than 90%of patients had clinical characteristics such as persistent irregular high fever,serous cavity effusion,decreased blood cells,and liver function dam-age.The levels of TH17,TH17/Treg,CD3+,and CD8+T cells on the second day of admission,1 month,2 months,and 3 months after treatment in the survival group were significantly lower than those in the fatality group(P<0.05),while the levels of Treg,CD4+,and CD16+CD56+lymphocytes were the opposite(P<0.05).The area under the reciever operating characteristics curve(AUCROC)of TH17/Treg cells combined with lymphocyte subsets in predicting the fatality of adult sHLH patients was 0.853(95%CI:0.758-0.922),which was higher than that of each individual indicator(P<0.05).Conclusion The clinical characteristics of adult sHLH are complex and diverse,with significant immune dysfunction of TH17/Treg and lymphocyte subsets.The TH17/Treg cells combined with lympho-cyte subsets have high efficacy in predicting the prognosis of adult sHLH patients.
3.Effect of Zuogui Jiangtang Tongmai Prescription on Inflammatory Injury of Human Umbilical Vein Endothelial Cells Induced by High Glucose and LPS Based on GPR43/β-arrestin-2/IκBα/NF-κB Pathway
Lanyu PENG ; Jingxin YAO ; Yujia LI ; Dingxiang LI ; Xun LIU ; Yihui DENG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(3):64-74
ObjectiveTo investigate the effects and mechanism of Zuogui Jiangtang Tongmai prescription (ZJTP) on human umbilical vein endothelial cells (HUVECs) damaged by high glucose combined with lipopolysaccharide (LPS). MethodThe survival rate of cells was determined by cell counting kit-8 (CCK-8), and the level of tumor necrosis factor-α (TNF-α) was determined by enzyme-linked immunosorbent assay (ELISA) to determine the optimal injury concentration and action time of LPS, as well as the optimal action concentration of ZJTP drug-containing serum. HUVECs were divided into a blank control group, a model group, a ZJTP drug-containing serum group, and an SCFA mixed liquid group. ELISA was used to detect the level of endothelin-1 (ET-1), nitric oxide (NO), interleukin-1β (IL-1β), interleukin-6 (IL-6), and TNF-α. Western blot was performed to detect the protein expression of G protein-coupled receptor43 (GPR43), β-suppressor protein-2 (β-arrestin-2), nuclear factor-κB suppressor α (IκBα), and nuclear factor κB p65 (NF-κB p65). The nucleation of NF-κB p65 was observed by immunofluorescence staining (IF). The role of GPR43 in the regulation of inflammatory injury was observed by means of small interfering ribonucleic acid (siRNA). The cells after intervention were divided into an empty carrier group, a ZJTP drug-containing serum group, a Si-GPR43 group, and a Si-GPR43 + ZJTP drug-containing serum group. The content of IL-1β, IL-6, and TNF-α was detected by ELISA. The protein expression of pathways was detected by Western blot. IF was used to observe the nucleation of NF-κB p65. ResultThe optimal molding condition was 1 mg·L-1 LPS for 24 h. The optimal drug intervention condition was 5% ZJTP drug-containing serum for 24 h. Compared with the blank control group, the content of ET-1 in the model group was significantly increased, and the content of NO was significantly decreased (P<0.01). The levels of inflammatory factors were significantly increased (P<0.01). The expressions of GPR43 and IκBα were significantly decreased, while the protein expressions of β-arrestin-2 and NF-κB p65 were significantly increased (P<0.01). NF-κB p65 protein was transferred from the extranuclear to the intranuclear (P<0.01). Compared with the model group, the content of ET-1 in the ZJTP drug-containing serum group was decreased, and the content of NO was increased (P<0.05). The levels of inflammatory factors decreased (P<0.05). The protein expressions of GPR43 and IκBα were increased, while the expressions of β-arrestin-2 and NF-κB p65 were decreased (P<0.05). The amount of NF-κB p65 transferred from the intranuclear to the extranuclear decreased (P<0.01). The mechanism study showed that compared with the Si-GPR43 group, the content of IL-1β, IL-6, and TNF-α were significantly decreased after treatment with ZJTP drug-containing serum (P<0.01). The protein expressions of GPR43 and IκBα were significantly increased (P<0.01), while the protein expressions of β-arrestin-2 and NF-κB p65 were significantly decreased (P<0.01). The amount of NF-κB p65 transferred from the extranuclear to the intranuclear decreased (P<0.01). ConclusionZJTP has a protective effect on HUVECs with high glucose and LPS-induced inflammatory injury, which may be related to the regulation of GPR43/β-arrestin-2/IκBα/NF-κB pathway.
4.Validity and reliability of the Chinese Social Anxiety Questionnaire for Adults
Yuxi WANG ; Yinyin ZANG ; Yujia PENG
Chinese Mental Health Journal 2024;38(8):730-736
Objective:To test the validity and reliability of the Chinese Social Anxiety Questionnaire for A-dults(CSAQ-A).Methods:Totally 555 participants[aged 18 to 60 years,average age(29±8)years old]were recruited to complete the CSAQ-A.The Liebowitz Social Anxiety Scale(LSAS),Brief Fear of Negative Evaluation Scale(BFNE),Penn State Worry Questionnaire(PSWQ),State-Trait Anxiety Inventory-Trait(STAI-T),and Beck Depression Scale-Second Edition(BDI-Ⅱ)were conducted to test the criterion validity.After 4 weeks,143 subjects were retested.Results:Confirmatory factor analysis demonstrated that the five-dimensional structure of the CSAQ-A exhibited excellent fit indices(x2/df=3.06,CFI=0.92,TLI=0.91,RMSEA=0.06,SRMR=0.05).The CSAQ-A scores were positively correlated with the scores of LSAS,BFNE,PSWQ,STAI-T and BDI-Ⅱ(ICC=0.40-0.67,Ps<0.001).The Cronbach α coefficient of the CSAQ-A scale was 0.96,and the test-retest reliability was 0.80.Conclusion:The Chinese Social Anxiety Questionnaire for Adults has good validity and reliability and could effectively evaluate social anxiety in the Chinese adult population.
5.The relationship between emotional intelligence and emotional labor in medical staff: the mediating role of humanistic caring ability
Hao YANG ; Wentao PENG ; Suxia WANG ; Hong ZHANG ; Jihong PAN ; Hui ZHU ; Yujia GUAN ; Wanying LUO
Chinese Journal of Medical Education Research 2023;22(11):1739-1744
Objective:To understand the status quo of emotional intelligence, emotional labor, and humanistic caring ability of medical staff, and to clarify their internal relationship.Methods:Convenience sampling was used to select 713 medical staff from a grade A tertiary hospital in Chengdu, Sichuan Province, China. Emotional Intelligence Scale, Humanistic Caring Scale, and Emotional Labor Scale were used to measure the emotional intelligence, humanistic caring ability, and emotional labor of medical staff. SPSS 22.0 software was used to establish a database for statistical description and analysis. Process 3.2 software was used to analyze the mediating effect.Results:In humanistic caring ability, the average score of comprehension dimension was the highest (75.62±8.20) and the average score of patience dimension was the lowest (58.53±5.01). In emotional labor, the average score of the deep action dimension was the highest (23.39±3.85) and the average score of the surface action dimension was the lowest (17.73±3.18). In emotional intelligence, the average score of self-emotion evaluation dimension was the highest (21.76±3.30) and the average score of other-emotion evaluation dimension was the lowest (20.07±3.71). Positive correlations were found between humanistic caring ability and emotional intelligence, between humanistic caring ability and emotional labor, and between emotional intelligence and emotional labor ( P<0.01). Humanistic caring ability had a partial mediating effect between emotional intelligence and emotional labor. Humanistic caring ability had direct and indirect effects on emotional labor, and the effect sizes were 0.279 and 0.029, respectively. Conclusion:Emotional intelligence has a direct positive predictive effect on emotional labor, humanistic caring ability as an intermediary variable indirectly and positively predicts emotional labor. Humanistic caring ability plays a partial mediating role between emotional intelligence and emotional labor. Attention should be paid to the emotional labor of medical staff, and the emotional labor of medical staff should be improved through targeted training on emotional intelligence and humanistic caring ability. These efforts will improve the current situation and establish a harmonious doctor-patient relationship.
6.Gender differences in mortality following tanscatheter aortic valve replacement (TAVR): a single-centre retrospective analysis from China.
Qi LIU ; Yali WANG ; Yijian LI ; Tianyuan XIONG ; Fei CHEN ; Yuanweixiang OU ; Xi WANG ; Yijun YAO ; Kaiyu JIA ; Yujia LIANG ; Xin WEI ; Xi LI ; Yong PENG ; Jiafu WEI ; Sen HE ; Qiao LI ; Wei MENG ; Guo CHEN ; Wenxia ZHOU ; Mingxia ZHENG ; Xuan ZHOU ; Zhengang ZHAO ; Chen MAO ; Feng YUAN
Chinese Medical Journal 2023;136(20):2511-2513
7.Peri-procedural myocardial injury predicts poor short-term prognosis after TAVR: A single-center retrospective analysis from China.
Qi LIU ; Kaiyu JIA ; Yijun YAO ; Yijian LI ; Tianyuan XIONG ; Fei CHEN ; Yuanweixiang OU ; Xi WANG ; Yujia LIANG ; Xi LI ; Yong PENG ; Jiafu WEI ; Sen HE ; Qiao LI ; Wei MENG ; Guo CHEN ; Wenxia ZHOU ; Mingxia ZHENG ; Xuan ZHOU ; Yuan FENG ; Mao CHEN
Chinese Medical Journal 2023;136(24):3013-3015
8.Effect of Zuogui Jiangtang Tongmai Prescription on Astrocyte Injury by AGEs Combined with Oxygen-glucose Deprivation Based on AMPK/mTOR/ULK1 Pathway Related to Autophagy
Yujia LI ; Dingxiang LI ; Xun PENG ; Yihui DENG
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(16):90-99
ObjectiveTo explore the effect and mechanism of Zuogui Jiangtang Tongmai prescription (ZGJTTMP) on astrocytes (ASs) injured by advanced glycation end products(AGEs) combined with oxygen-glucose deprivation (OGD). MethodCell counting kit-8 (CCK-8) was used to determine the optimal concentration of AGEs and the action time of OGD, and the optimal blood concentration of ZGJTTMP was selected for follow-up experiments. ASs were divided into normal group, model group (AGEs + OGD), ZGJTTMP group, an adenosine 5'-monophosphate-activated protein kinase (AMPK) inhibitor (Compound C) group, AMPK activator (AICAR) group, and combination group (ZGJTTMP + AICAR). The morphological changes in ASs in each group were observed under an inverted microscope. The cell survival rate in each group was detected by CCK-8. The content of interleukin-1β(IL-1β), interleukin-6(IL-6), and tumor necrosis factor-α(TNF-α) was detected by enzyme-linked immunosorbent assay (ELISA). The number of autophagosomes in each group was counted under an electron microscope. The expression of microtubule-associated protein light chain 3 (LC3) was observed by immunofluorescence. The protein expression of LC3, p62, p-AMPK, AMPK, p-mammalian target of rapamycin (mTOR), mTOR, p-UNC-51 like kinase 1 (ULK1), and ULK1 was detected by Western blot. ResultAccording to the results of cell survival rate, 200 mg·L-1 AGEs and OGD for 6 h were selected as the optimal modeling conditions for the model group, and 5% was selected as the optimal blood concentration of ZGJTTMP. Under the inverted microscope, the cells were severely damaged after modeling, but the cell injury in the ZGJTTMP group and the Compound C group was significantly improved. As revealed by ELISA results, the content of IL-1β, IL-6, and TNF-α in the model group increased (P<0.01), and the content of inflammatory factors in the ZGJTTMP group and the Compound C group decreased (P<0.01). Under the electron microscope, the number of autophagosomes in the model group increased significantly. The immunofluorescence results showed that the expression area of LC3 increased in the model group (P<0.01), and the ZGJTTMP group and the Compound C group showed decreased number of autophagosomes and reduced expression area of LC3 (P<0.01). As demonstrated by the results of Western blot, compared with the normal group, the model group showed increased expression of LC3Ⅱ/LC3Ⅰ and p-AMPK/AMPK (P<0.01) and decreased p62, p-mTOR/mTOR, and p-ULK1/ULK1 (P<0.01). Compared with the model group, the ZGJTTMP group and the Compound C group showed decreased expression of LC3Ⅱ/LC3Ⅰ and p-AMPK/AMPK (P<0.01) and increased p62, p-mTOR/mTOR, and p-ULK1/ULK1 (P<0.01). ConclusionZGJTTMP possesses a protective effect on ASs with inflammatory injury by AGEs combined with OGD, which may be achieved by inhibiting the activation of the AMPK/mTOR/ULK1 pathway related to autophagy, thus inhibiting the overexpression of autophagy.
9.Construction of undergraduate teaching method of standardized ultrasound skill operation video and evaluation of its teaching effect
Ying HE ; Jingyi ZHANG ; Yujia YANG ; Yue ZHANG ; Yulan PENG
Chinese Journal of Medical Education Research 2022;21(7):858-862
This study evaluated the standardized ultrasound operation skill video courseware recording and online teaching of undergraduate clinical internship course in the direction of medical imaging ultrasound technology in West China Clinical Medical College of Sichuan University. Through the systematic teaching plan preparation of ultrasound skill operation teaching video, script writing, post production, recording and implementation of teaching, the results showed that 116 students (81.69%) thought knowledge points of skill operation video course were reasonably divided, 128 ones (90.14%) thought the explanation was clear, 118 ones (83.10%) thought the video duration was appropriate, 121 ones (85.21%) thought the degree of difficulty was moderate, 119 ones (83.09%) thought the ultrasonic image was consistent with the anatomical image, and 124 ones (87.32%) thought it was helpful to improve the level of clinical operation skills. Standardized ultrasound skill operation video online teaching has the advantages of no time and place restrictions and good repeatability, which can exercise students' self-study ability, improve their learning enthusiasm, help students master operation skills more comprehensively, and is of great significance to the implementation and promotion of ultrasound clinical skill training video online teaching.
10.Metabolic outcomes of type 2 diabetes patients with different diabetic durations under the standardized metabolic disease management model
Jiaying YANG ; Yujia GONG ; Mengyu LAI ; Na LI ; Aifang ZHANG ; Liping GU ; Yufan WANG ; Yongde PENG
Chinese Journal of Endocrinology and Metabolism 2021;37(2):106-113
Objective:To explore the metabolic outcomes of type 2 diabetes patients with different durations after 1 year treatment under the standardized metabolic disease management model.Methods:(1)From September 2017 to September 2018, 345 type 2 diabetes patients in the Standardized Metabolic Management Center(MMC) of Shanghai General Hospital were recruited and included in this research. They were divided into newly-diagnosed type 2 diabetes(duration≤1 year) and long-term groups(duration>1 year). The general characteristics, blood pressures, glycemic levels, lipids levels, control rates and comprehensive compliance rates(blood glucose, pressure and lipids all reached targets) were compared at baseline between 2 groups.(2)All patients underwent one year standardized management, and metabolic indicators mentioned above and control rates at the time were compared as well.Results:(1) At baseline, compared with long-term group, patients in newly-diagnosed type 2 diabetes group were younger ( P<0.01), and 2 h blood glucose level after glucose loading were higher [(15.20±5.26 vs 13.68±4.94) mmol/L, P<0.01]. (2) After one year standardized management, body weight, blood pressure, glucose and lipids metabolism in all patients were significantly improved. Compared with patients in long-term group, newly-diagnosed type 2 diabetes patients achieved better glycemic level [fasting blood glucose(6.27±1.56 vs 7.63±2.08) mmol/L, P<0.01; glycated hemoglobin(6.33±0.96 vs 7.23±1.37) %, P<0.01] , and had higher HOMA-β [(74.01±56.45 vs 40.17±37.07) %, P<0.01]. The glycemic control, blood pressure and blood lipids control rates in both groups increased significantly in one year. Comprehensive compliance rate of the whole patients increased from 5.80% to 24.06%. The metabolism indexes of the newly-diagnosed type 2 diabetes group were better than those of the long-term group[comprehensive compliance rate: (24.73% vs 17.18%, P=0.087, glycemic control rate(84.62% vs 53.37%, P<0.01)]. Conclusion:Standardized metabolic disease management promoted the overall improvement in blood glucose, blood pressure, and lipids levels in type 2 diabetes patients, especially in terms of blood glucose and those of the newly-diagnosed type 2 diabetes. In the future, we should focus on the early diagnosis and treatment of type 2 diabetes, actively promote the MMC model and stress the integrated management of blood glucose, blood pressure, and blood lipid levels. We should pay more attention to the long-term patients, to improve their awareness and treatment compliance.

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