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Equipment for screening and analysis of target substances in the complex system of traditional Chinese medicine based on cell membrane chromatography technology

Langchong HE ; Huaizhen HE ; Shengli HAN ; Tao ZHANG ; Cheng WANG ; Weina MA

Journal of Xi'an Jiaotong University(Medical Sciences).2024;45(3):351-359. doi:10.7652/jdyxb202403001

Traditional Chinese medicine(TCM)is extensively utilized for clinical disease prevention and treatment.However,due to the intricate nature of its material basis and the multiple factors involved in the preparation process,ensuring comprehensive quality control of TCM proves to be challenging.By instilling a clear understanding of its effective and harmful substances and implementing control over the content and limit of TCM during the preparation process,the controllability and repeatability of its quality can be guaranteed.Currently,China is facing a dearth of innovative technology for drug development,necessitating an increase in research and development efficiency,especially in the realm of high-throughput precision analytical equipment.The country has long relied on imported pharmaceutical analysis equipment with a particular efficiency in high-end intelligent analysis equipment.This is especially concerning considering the urgent requirement to establish a"pharmaceutical intelligent analysis system."This project,supported by the Major Instrument Development Project of the National Science and Technology Funds,employs cell membrane chromatography technology,complemented by biotechnology and artificial intelligence technology,to devise a two-dimensional cell membrane chromatography(2D/CMC)analyzer.The project has successfully conducted a demonstration application of the"2D/CMC-Traditional Chinese Medicine Pharmacodynamic Substance Analyzer"and the"2D/CMC-Traditional Chinese Medicine Injection Allergen Analyzer".These tools have enhanced the screening and discovery efficiency of TCM's effective substances and allergen components.Moreover,the equipment amalgamates qualitative and quantitative analysis,thereby serving as an effective analytical tool to enhance the quality and efficacy of traditional Chinese medicine.

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Opioid-induced fragile regulatory T cells contribute to NAc synaptic plasticity and withdrawal symptoms

Yin LOU ; Tianyou MA ; Yongsheng ZHU

Journal of Xi'an Jiaotong University(Medical Sciences).2024;45(3):360-365. doi:10.7652/jdyxb202403002

Decline of immunity is an epidemiological feature of opioid addicts.Recent work reveals a landscape of peripheral immune microenvironment in opioid addicts.Opioid addicts exhibit a significant expansion of fragile-like regulatory T cells(Tregs)and enhanced Treg-derived interferon-y(IFN-γ)expression.IFN-γ signaling reshapes synaptic morphology in nucleus accumbens(NAc)neurons,modulating subsequent withdrawal symptoms.Treg fragility transformation from WT Tregs is primarily due to opioid-induced global hypoxia during acute withdrawal period.Opioids increase the expression of neuron-derived C-C motif chemokine ligand 2(Ccl2)and disrupt blood-brain barrier(BBB)integrity through the downregulation of astrocyte-derived fatty-acid-binding protein 7(Fabp7),both of which trigger peripheral Treg infiltration into NAc.Recent studies suggest that subtle homeostatic changes in the peripheralimmune milieu may also contribute to modulating synapses that are responsible for addictive behaviors,which may lead to the development of new therapeutic strategies.

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Reliability,validity,and cut-off score of the Chinese version of the Suicidal Behaviors Questionnaire-Revised

Caizhi WU ; Xin BAI ; Xueyu PENG ; Lixia YU ; Zhihong REN

Journal of Xi'an Jiaotong University(Medical Sciences).2024;45(3):366-370. doi:10.7652/jdyxb202403003

Objective To examine the reliability and validity of the Suicidal Behaviors Questionnaire-Revised(SBQ-R)among Chinese undergraduates and to find out cut-off score for screening the risk of suicide in clinical undergraduates.Methods A total of 488 Chinese undergraduates participated in this study.Of them,366 participants were asked to complete the Chinese version of SBQ-R,Psychache Scale,and Satisfaction with Life Scale.And 45 undergraduates were re-tested after four weeks;122 undergraduates were tested by using the Suicidality Module Section of the Mini International Neuropsychiatric Interview and the Chinese version of SBQ-R.Results ① The results showed that The Chinese version of SBQ-R had one factor with good fit indices.② The internal consistency reliability,split-half reliability,and one-month retest reliability of the total questionnaire were 0.76,0.79,and 0.93,respectively.③ The total score of the SBQ-R in Chinese was positively correlated with psychache but negatively correlated with satisfaction of life,indicating that the scale has good criterion-related validity.④ Receiver operating characteristics(ROC)curve evaluation suggested that the cut-off score was 9 in terms of identifying the clinical undergraduates at high risk of suicide.Conclusion SBQ-R is a reliable,valid,and practical instrument to measure the risk of suicide among Chinese undergraduates.SBQ-R＞9 is the best cut-off value for screening for high suicide risk in China.

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Relationship between college students'perceived stress and their suicidal ideation:multiple mediating effect of resilience and depression

Caizhi WU ; Na MA ; Tingting HUANG ; Afang LU ; Zhihong REN ; Fenghui YUAN

Journal of Xi'an Jiaotong University(Medical Sciences).2024;45(3):371-375. doi:10.7652/jdyxb202403004

Objective To explore the relationship between perceived stress and suicidal ideation as well as the mediating effect of resilience and depression.Methods A sample of 875 college students was recruited for the study to complete a set of self-report measures of Chinese Perceived Stress Scale(CPSS),the Resiliency Scale of University Students,Suicidal Ideation Questionnaire,and Beck Depression Inventory(BDI).Results ①College students'perceived stress and depression had significantly positive correlation with their suicidal ideation,while the students'resilience had a significantly negative correlation with their perceived stress and depression.②Perceived stress did not affect suicidal ideation of the college students directly,but indirectly affected their suicidal ideation through three significant mediating pathways:the separate mediating effects of(a)resilience and(b)depression,and the serial mediating effect of(c)resilience and depression.Conclusion Resilience and depression exert a chain mediating effect between perceived stress and suicidal ideation of college students.

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Impact of emotions on cancer risk:a Mendelian randomization study

Qin DU ; Yuxuan ZHU ; Yize GUO ; Nanxin XU ; Di LIU

Journal of Xi'an Jiaotong University(Medical Sciences).2024;45(3):376-382. doi:10.7652/jdyxb202403005

Objective To investigate the effects of emotions(subjective well-being,depressed effect,worry,and guilt)on cancer(colorectal cancer,hepatic cancer,thyroid cancer,lung cancer,and breast cancer).Methods Two-sample bi-directional Mendelian randomization(MR)method was adopted.All data were based on summary data from genome-wide association studies(GWAS).Inverse variance weighting(IVW)was used to generate the main results,and weighted median(WM)and MR-Egger methods were employed to calculate supplementary results.The outcome measure was odds ratio(OR),and sensitivity analysis was conducted.Results For depressed effect,a significant association with lung cancer(OR=1.005,95％CI:1.001-1.009,P=0.015)was found.For worry,a significant association with breast cancer(OR=1.199,95％CI:1.011-1.423,P=0.038)was observed.For guilt,a significant association with thyroid cancer(OR=2.083,95％CI:1.080-4.017,P=0.029)was identified.After removing all potentially pleiotropic SNPs detected by MR PRESSO,the association between worry and breast cancer showed no statistical difference(P=0.064),while the association between worry and colorectal cancer remained significant(OR=0.739,95％CI:0.571-0.956,P=0.021).No causal relationship was found between cancer and emotions.Conclusion There is a causal relationship between depression and increased lung cancer incidence,guilt and increased thyroid cancer incidence,as well as anxiety and decreased colorectal cancer incidence.

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Effect of fisetin against venous thrombosis in rats and its mechanism

Lihui LONG ; Shuang WEI ; Qing LIU ; Yang YAO ; Juanni DONG ; Yuanyuan CHANG ; Enhui WEN

Journal of Xi'an Jiaotong University(Medical Sciences).2024;45(3):383-387. doi:10.7652/jdyxb202403006

Objective To analyze the effect of fisetin against venous thrombosis in rats.Methods Seventy SD rats were randomly divided into the following groups:sham-operation group,model group,fisetin 45 mg/kg,15 mg/kg,5 mg/kg groups,and aspirin group(47 mg/kg).The corresponding medication was administered by gavage once a day consecutively(the sham-operation group and the model group were given 0.5％carboxymethyl cellulose sodium solution with 10 mL/kg,respectively)for 7 consecutive days.One hour after the last administration,the rats were anesthetized,the lower part of the intersection of inferior vena cava and left renal vein was ligated with silk thread(no ligation in the sham-operation group),and the abdominal wall was sutured.Two hours later,the abdominal cavity was reopened,the other venous branches 1.5 cm away from the ligation site were closed with the artery clamp,and blood was collected from the abdominal aorta.The anticoagulant ratio of 3.8％sodium citrate∶whole blood was 1∶9.The venous thrombus 1 cm down from the ligation point of the intersection of inferior vena cava and left renal vein was cut and the thrombus was separated.The residual blood was dried with filter paper,weighed and recorded.Plasma was taken after anticoagulant blood centrifugation.The levels of plasma antithrombin-Ⅲ(AT-Ⅲ),protease C(PC),plasminogen(PLG),and plasminogen activator inhibitor(PAI-1)were detected by ELISA kits.Results Compared with the model group,the weight of thrombus in fisetin 45 mg/kg group and aspirin 47 mg/kg group decreased(P＜0.01).The content of AT-Ⅲ in three fisetin groups increased(all P＜0.05).The content of PC in fisetin 45 mg/kg increased(P＜0.05).The content of PLG and PAI-1 in fisetin 45 mg/kg group decreased(both P＜0.05).Conclusion Fisetin has the effect against venous thrombosis in vivo,and the effect is related to the upregulation of AT-Ⅲ and PC and the downregulation of PLG and PAI-1.

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Effects of low temperature plasma-activated medium on proliferation and angiogenic capacity of vascular endothelial cells

Wang YUAN ; Xiangni WANG ; Jinren LIU ; Xiying WANG ; Jiajia LU ; Zhirou HE ; Yulin XU ; Xingmin SHI

Journal of Xi'an Jiaotong University(Medical Sciences).2024;45(3):388-395. doi:10.7652/jdyxb202403007

Objective To explore the plasma-activated medium(PAM)produced by low temperature plasma(LTP)on the proliferation and angiogenesis of human umbilical vein endothelial cells(HUVECs)so as to provide theoretical basis for the future use of PAM to promote wound healing and inhibit tumor angiogenesis.Methods HUVECs were selected as the in vitro research model.The PAM-containing medium after LTP treatment for different time points(0 s,15 s,30 s,45 s,60 s,and 75 s)was used for intervention.The influence of PAM on HUVECs viability was assessed using the MTT assay and cell cycle analysis.The effects of PAM on angiogenesis were examined through angiogenesis experiments.Intracellular levels of reactive oxygen species(ROS)were measured using fluorescence probes.A melanoma mouse model was established,and CD31 expression was detected by immunohistochemistry.Results As the treatment time increased,the intracellular levels of ROS also elevated.PAM derived from LTP exhibited a bidirectional effect on angiogenesis in HUVECs.Compared to the control group(0 s),low-dose treatments(15 s and 30 s)enhanced HUVECs viability,while high-dose treatments(45 s,60 s,and 75 s)significantly decreased cell viability(P＜0.05).The proportion of HUVECs in the S phase was significantly increased in the PAM-15 s and PAM-30 s groups,but markedly decreased in the PAM-45 s,PAM-60 s,and PAM-75 s groups,with statistically significant differences(P＜0.05).The HUVECs tube formation ability was enhanced in the 15 s and 30 s PAM groups,but diminished in the PAM-45 s,PAM-60 s,and PAM-75 s groups,characterized by the decreased numbers of vascular nodes,intersections,meshes,and branching points(P＜0.05).After PAM treatment in the melanoma mouse model,the control group exhibited widespread distribution of CD31 in tumor tissue,while the PAM-5 min and PAM-10 min groups displayed reduced distribution of CD31.Conclusion Short-term exposure to PAM enhances HUVECs proliferation and angiogenesis,whereas prolonged exposure suppresses cell viability and inhibits angiogenesis.

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Extracellular matrix stiffness induces CCL5 synthesis to enhance immunotherapy response in non-small cell lung cancer

Tian ZHANG ; Yaru YANG ; Enyong ZHANG ; Hui GUO ; Mengjie LIU

Journal of Xi'an Jiaotong University(Medical Sciences).2024;45(3):396-404. doi:10.7652/jdyxb202403008

Objective To investigate the mechanism of C-C motif chemokine ligand 5(CCL5)modulation by extracellular matrix stiffness in non-small cell lung cancer(NSCLC)and to determine the effect of CCL5 on the immunotherapy response of NSCLC.Methods The correlation between extracellular matrix stiffness and CCL5 expression in NSCLC was analyzed with the TCGA database.Polyacrylamide hydrogels with different stiffness were designed according to the extracellular matrix stiffness of NSCLC,and H1299 cells responding to the mechanical loading of hydrogel matrix stiffness were subjected to transcriptome sequencing.High matrix stiffness was verified to promote the expression of CCL5 by using sequence.Results High extracellular matrix stiffness upregulated CCL5 expression,and interferon-γ mediated signaling pathway might be involved in the process.NSCLC patients with high CCL5 expression had a greater abundance of cytotoxic T-cells in tumor tissue and reacted favorably to anti-programmed cell death protein 1 treatment.Conclusion Increased extracellular matrix stiffness promotes CCL5 synthesis,and CCL5 enhances immunotherapy responsiveness in NSCLC by increasing cytotoxic T cell infiltration in tumor tissue.

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Effect of selinexor combined with imatinib on proliferation and apoptosis of K562/G01 cells

Xiaojing HAO ; Liangming MAO

Journal of Xi'an Jiaotong University(Medical Sciences).2024;45(3):405-410. doi:10.7652/jdyxb202403009

Objective To observe the effect of Selinexor(SEL)combined with Imatinib(IM)on the proliferation and apoptosis of Imatinib-resistant chronic myeloid leukemia K562/G01(KG)cells and explore the possible mechanisms.Methods K562 cells and KG cells were treated with SEL or IM respectively or in combination.Cells viability was examined by MTT assay.Apoptosis was assessed by flow cytometry.BCR-ABL mRNA was detected by RT-PCR.XPO1 was detected by Western blotting.Results IM and SEL both inhibited the proliferation of K562 cells and KG cells;half maximal inhibitory concentration(IC50)for 48 h was 0.16 μmol/L vs.6.48 μmol/L for IM and 132.0 nmol/L vs.275.9 nmol/L for SEL.Compared with SEL or IM alone,SEL combined with IM significantly inhibited the proliferation of KG cells(P＜0.05),induced KG cells apoptosis(P＜0.05),downregulated the levels of BCR-ABL mRNA(P＜0.05),and inhibited the expressions of XPO1 in KG cells(P＜0.05).Conclusion SEL combined with IM can synergistically inhibit the proliferation and induce apoptosis of KG cells,and then inhibit the expressions of BCR-ABL mRNA and XPO1 to exert an anti-leukemia effect.

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The molecular mechanism of GSDMB regulating the fate of intestinal epithelial cells

Jun YANG ; Zhengrui DENG ; Yi LI ; Sen WU ; Kun GUO ; Wenbin GONG

Journal of Xi'an Jiaotong University(Medical Sciences).2024;45(3):411-418. doi:10.7652/jdyxb202403010

Objective To explore the molecular mechanism of Gasdermin B(GSDMB)regulating the fate of intestinal epithelial cells.Methods The human GSDMB plasmid was overexpressed into two human intestinal epithelial cell lines(NCM460 and HT-29 cells)and human colon-derived organoids.Western blotting was used to confirm the efficiency of electroporation.Cell counting kit(CCK8),cell apoptosis,and cell cycle by flow cytometry were performed to analyze the effect of GSDMB overexpression on cell function.Transcriptome sequencing was used to analyze the downstream effector molecules of GSDMB.T test was used to compare the data between the two groups.Results The overexpression of GSDMB protein in the two intestinal epithelial cell lines was successfully reconstructed.The absorbance value(A)of human intestinal epithelial cells overexpressing GSDMB protein[NCM460 cells:(1.17±0.01),HT-29 cells:(0.96±0.06)]was significantly lower than that of blank control cells[NCM460 cells:(1.67±0.12),HT-29 cells:(1.24±0.07)](t=7.24 and 5.46,P＜0.05).The number of apoptotic cells in the GSDMB overexpression group[NCM460 cells:(12.03±1.55),HT-29 cells:(29.30±4.48)]was significantly higher than that in the blank group[NCM460 cells:(4.96±1.74),HT-29 cells:(6.95±3.42)](t=5.26 and 6.97,P＜0.05).Cell cycle analysis showed that the ratio of cells at G0/G1 phase in the GSDMB overexpression group[NCM460 cells:(47.98±5.28)％,HT-29 cells:(38.04±3.45)％]was significantly lower than that in the control group[NCM460 cells:(59.54±3.90)％,HT-29 cells:(63.81±1.76)％](t=3.05 and 11.53,P＜0.05).Transcriptome sequencing results showed that the dual specificity phosphatase 4 and 6(DUSP4 and DUSP6)genes were significantly upregulated after GSDMB protein expression.Fluorescence quantitative PCR results confirmed that the relative expression levels of DUSP4(2.45±0.15)and DUSP6(4.34±0.22)in intestinal epithelial cells transfected with GSDMB were significantly higher than those in the control group(1.06±0.05 and 1.01±0.02)(t=15.08 and 26.52,P＜0.05).After GSDMB-expressing NCM460 cells were treated with the DUSP inhibitor BCI,the BCI treatment group had a significantly increased expression level of p-ERK compared to the control group[(1.14±0.17)vs.(0.58±0.12)](t=5.42,P=0.002);the A value(1.84±0.07)and G0/G1 phase ratio(59.83±2.17)％in the BCI treatment group were significantly higher than those in the non-treatment group[(1.52±0.10)and(52.10±2.23)％],and the number of apoptosis in the BCI treated group(7.60±0.56)was significantly lower than that in the untreated group(12.57±1.00)(t=4.71,4.31,7.52,P＜0.05).TUNEL staining in human colon organoids showed a significant increase in apoptotic cells,and the relative expression level of DUSP6 protein(0.85±0.09)was significantly higher than that of the control group(0.21±0.04),accompanied by a decrease in p-ERK levels[(0.83±0.18)vs.(0.19±0.06)],with statistical significance(t=11.95,P＜0.001;t=6.56,P＜0.001).Conclusion GSDMB may inhibit cell proliferation,induce cell cycle arrest,and promote apoptosis by upregulating dual specificity phosphatase DUSP6-mediated ERK phosphorylation,thus affecting the fate of intestinal epithelial cells.

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