1.Exploring Mechanism of Hei Xiaoyaosan Regulating PI3K/Akt Pathway to Improve Learning and Memory Ability of Insomnia Rats with Liver Depression Syndrome Based on Transcriptomics
Jiamin LIU ; Yale WANG ; Hai HUANG ; Yue LI ; Xin FAN ; Pengpeng LIANG ; Shizhao ZHANG ; Mei YAN ; Guiyun LI ; Hongyan WU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(16):114-125
ObjectiveBased on transcriptomics, to explore the mechanism of Hei Xiaoyaosan regulating the phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) signaling pathway to improve the learning and memory ability of insomnia rats with liver depression syndrome. MethodsSixty 8-week-old male SD rats were randomly divided into the blank group, model group, eszopiclone group (0.09 mg·kg-1), and low, medium, and high dose groups of Hei Xiaoyaosan (3.82, 7.65, 15.30 g·kg-1), with ten rats in each group. Except for the blank group, the other groups were induced insomnia rat model with liver depression by chronic restraint, tail clamping stimulation and intraperitoneal injection of p-chlorophenylalanine (PCPA). Each treatment group received intragastric administration according to the specified dosage, once a day for 14 consecutive days. The pentobarbital sodium cooperative sleep test, open field test, and Morris water maze test were used to test the sleep quality, depressive-like behavior, and learning and memory abilities of rats. Additionally, enzyme-linked immunosorbent assay (ELISA) was used to detect the contents of 5-hydroxytryptamine (5-HT), γ-aminobutyric acid (GABA), brain-derived neurotrophic factor (BDNF), glial cell line-derived neurotrophic factor (GDNF) and nitric oxide (NO) in hippocampus. Hematoxylin-eosin (HE) staining was performed to observe pathological changes of the hippocampal tissue, while terminal deoxynucleotidyl transferase deoxyuridine triphosphate (dUTP) nick end labeling (TUNEL) was used to evaluate apoptosis of hippocampal neurons. Transcriptomic sequencing technology was employed to identify differentially expressed genes in hippocampus between the model group and the blank group, as well as between the medium-dose group of Hei Xiaoyaosan and the model group. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis were performed on the intersecting genes. Subsequently, the enriched key genes and signaling pathways were analyzed and verified. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was utilized to assess the mRNA expression levels of phosphatase and tensin homolog (PTEN), B-cell lymphoma-2 (Bcl-2)-like protein 11 (BCL2L11), and mitogen-activated protein kinase 1 (MAPK1) in hippocampus, and Western blot was employed to evaluate the protein expressions of PI3K, phosphorylation (p)-PI3K, Akt, p-Akt, Bcl-2, Bcl-2-associated X protein (Bax), and cleaved Caspase-3 in the same tissue. ResultsCompared with the blank group, the model group exhibited a reduction in body weight, an increase in sleep latency, and a decrease in sleep duration (P<0.01). Additionally, rats showed obvious depression-like behavior, and their learning and memory abilities decreased. Furthermore, the contents of 5-HT, GABA, NO, BDNF and GDNF in hippocampus decreased (P<0.01). Histological examination revealed a disorganized cell arrangement in the CA1 region of the hippocampus, characterized by irregular cell shapes, a reduced cell count, deeply stained and pyknotic nuclei, increased vacuolar degeneration, and an elevated apoptosis rate (P<0.01). Compared with the model group, the body weight of the high and medium dose groups of Hei Xiaoyaosan increased, the sleep latency shortened and the sleep time prolonged (P<0.05, P<0.01). Additionally, depression-like behavior and learning and memory abilities of rats were significantly improved, the levels of 5-HT, GABA, NO, BDNF and GDNF in the hippocampus increased (P<0.05, P<0.01). These interventions also ameliorated pathological damage in the hippocampal CA1 area and reduced the apoptosis of hippocampal neurons (P<0.01). Transcriptomic sequencing results indicated that Hei Xiaoyaosan might exert a therapeutic effect by regulating PI3K/Akt pathway through key mRNAs such as PTEN, BCL2L11, and MAPK1. The roles of these key mRNAs and proteins within PI3K/Akt pathway were further validated. In comparison to the blank group, the expression levels of PTEN, BCL2L11 and MAPK1 mRNA in the hippocampus of rats in the model group were increased (P<0.01), while the protein expression levels of p-PI3K, p-Akt and Bcl-2 were decreased (P<0.01), and the protein expression levels of PTEN, Bax and cleaved Caspase-3 were increased (P<0.01). Compared with the model group, the high-dose and medium-dose groups of Hei Xiaoyaosan could down-regulate the expressions of PTEN, BCL2L11 and MAPK1 mRNAs (P<0.01), up-regulate the expressions of p-PI3K, p-Akt and Bcl-2 proteins (P<0.01), and down-regulate the protein expressions of PTEN, Bax and cleaved Caspase-3 (P<0.05, P<0.01). ConclusionHei Xiaoyaosan may regulate PI3K/Akt signaling pathway by down-regulating expressions of key genes such as PTEN, BCL2L11 and MAPK1, and thus improve the learning and memory abilities of insomnia rats with liver depression syndrome.
2.Analysis of clinicopathological features, therapeutic efficacy, prognosis and mutation in 11 patients with cardiac diffuse large B-cell lymphoma
Na ZHAO ; Qing SHI ; Shu CHENG ; Li WANG ; Pengpeng XU ; Weili ZHAO
Journal of Leukemia & Lymphoma 2024;33(2):97-103
Objective:To investigate the clinicopathological features, mutation, therapeutic efficacy and the factors influencing the prognosis of patients with cardiac diffuse large B-cell lymphoma (DLBCL).Methods:A retrospective case series study was performed. The clinical data of 11 cardiac DLBCL patients in Ruijin Hospital of Shanghai Jiao Tong University School of Medicine from January 2016 to October 2020 were retrospectively analyzed. NovaSeq sequencing platform was used to detect gene mutations in 5 patients, and bioinformatics analysis of sequencing data was conducted through public database to identify the mutation sites of pathogenic genes. Kaplan-Meier method was used to analyze the progression-free survival (PFS) and the overall survival (OS). Univariate Cox proportional risk model was used to analyze the influencing factors of prognosis.Results:Among 11 patients with cardiac DLBCL, 5 were male and 6 were female. The age [ M ( Q1, Q3)] was 61 years (45 years, 70 years). All 11 patients were non-germinal center B-cell (non-GCB) type. There were 2 primary cases and 9 secondary cases; 9 cases with Ann Arbor stage of Ⅲ-Ⅳ, 10 cases with increased lactate dehydrogenase (LDH) and 9 cases with international prognostic index (IPI) score equal to or higher than 3 scores. Among 11 patients, 9 cases received a first-line treatment based on the R-CHOP (rituximab, cyclophosphamide, epirubicin/doxorubicin hydrochloride liposomes, vincristine and prednisone) regimen, of which 8 patients achieved complete remission (CR), and 1 patient achieved stable disease (SD); 1 patient received IR2 (ibrutinib + rituximab + lenalidomide) treatment regimen and achieved SD, and 1 patient received supportive treatment only and achieved progression of the disease. The follow-up time was 39.9 months (25.6 months, 57.3 months). The 3-year PFS rate and 3-year OS rate of 11 patients was 54.5%, 77.9 %, respectively. Univariate Cox regression analysis showed that gender, B symptoms, Ann Arbor stage, LDH level, number of extranodal lesions, IPI score were not correlated with PFS and OS of patients (all P > 0.05). Among 5 cases undergoing gene detection, KMT2D mutations and PIM1 mutations were detected in 2 cases,respectively. Interestingly, KMT2D mutations were only found in secondary cardiac DLBCL patients (2/3), while PIM1 mutations were only detected in primary cardiac DLBCL patients (2/2). Conclusions:Most cardiac DLBCL patients are non-GCB type and have advanced clinical stage, while may benefit from R-CHOP treatment regimen. PIM1 and KMT2D are the commonly mutated genes in cardiac DLBCL.
3.Clinicopathologic characteristics,gene mutation profile and prognostic analysis of thyroid diffuse large B-cell lymphoma
Zhishan DU ; Yue WANG ; Ziyang SHI ; Qing SHI ; Hongmei YI ; Lei DONG ; Li WANG ; Shu CHENG ; Pengpeng XU ; Weili ZHAO
Journal of Shanghai Jiaotong University(Medical Science) 2024;44(1):64-71
Objective·To analyze the clinicopathologic characteristics,gene mutation profile,and prognostic factors of thyroid diffuse large B-cell lymphoma(DLBCL).Methods·From November 2003 to December 2021,a total of 66 patients with thyroid DLBCL[23 cases(34.8%)with primary thyroid DLBCL,and 43 cases(65.2%)with secondary thyroid DLBCL]admitted to Ruijin Hospital,Shanghai Jiao Tong University School of Medicine were retrospectively analyzed for their clinicopathological data,survival and prognostic factors.Gene mutation profiles were evaluated by targeted sequencing(55 lymphoma-related genes)in 40 patients.Results·Compared to primary thyroid DLBCL,secondary thyroid DLBCL had advanced ratio of Ann Arbor stage Ⅲ?Ⅳ(P=0.000),elevated serum lactate dehydrogenase(LDH)(P=0.043),number of affected extranodal involvement≥2(P=0.000),non-germinal center B cell(non-GCB)(P=0.030),BCL-2/MYC double expression(DE)(P=0.026),and international prognostic index(IPI)3?5-scores(P=0.000).The proportion of patients who underwent thyroid surgery(P=0.012)was lower than that of patients with primary thyroid DLBCL.The complete remission(CR)rate in primary thyroid DLBCL patients was higher than that in secondary thyroid DLBCL patients(P=0.039).Fifty-five patients(83%)received rituximab combined with cyclophosphamide,doxorubicin,vincristine,and prednisone(R-CHOP)-based first-line regimen.The estimated 5-year progression free survival(PFS)rate of primary thyroid DLBCL patients was 95.0%,higher than the 49.7%of the secondary patients(P=0.010).Univariate analysis showed that Ann Arbor Ⅲ?Ⅳ(HR=4.411,95%CI 1.373?14.170),elevated LDH(HR=5.500,95%CI 1.519?19.911),non-GCB(HR= 5.291,95%CI 1.667?16.788),and DE(HR=6.178,95%CI 1.813?21.058)were adverse prognostic factors of PFS in patients with thyroid DLBCL.Ann Arbor Ⅲ?Ⅳ(HR=7.088,95%CI 0.827?60.717),elevated LDH(HR=6.982,95%CI 0.809?60.266),and DE(HR=18.079,95%CI 1.837?177.923)were adverse prognostic factors of overall survival(OS).Multivariate analysis showed that Ann Arbor Ⅲ?Ⅳ(HR=4.693,95%CI 1.218?18.081)and elevated LDH(HR=5.058,95%CI 1.166?21.941)were independent adverse prognostic factors of PFS in patients with thyroid DLBCL.Targeted sequencing data showed mutation frequency>20%in TET2(n=14,35%),KMT2D(n=13,32%),TP53(n=11,28%),GNA13(n=10,25%),KMT2C(n=9,22%),and TP53 were adverse prognostic factors of PFS in patients with thyroid DLBCL(P=0.000).Conclusion·Patients with primary thyroid DLBCL have better PFS and OS than those with secondary thyroid DLBCL.Ann Arbor Ⅲ?Ⅳ,elevated LDH,non-GCB,and DE(MYC and BCL2)are adverse prognostic factors in thyroid DLBCL.TET2,KMT2D,TP53,GNA13,and KMT2C are commonly highly mutated genes in thyroid DLBCL,and the prognosis of patients with TP53 mutations is poor.
4.Effect of culture time on immune-related membrane proteins of mouse dendritic cells and their exosomes
Li XIAO ; Shumin LUO ; Fang XU ; Pengpeng LU ; Enhong XING ; Weihua LI
The Journal of Practical Medicine 2024;40(7):941-947
Objective This study aims to investigate the impact of cultivation time on dendritic cells(DCs)and their derived exosomes′ expression of immune-related membrane proteins(CD80,MHC-Ⅰ,MHC-Ⅱ)and provides experimental evidence for future research.Methods Mouse bone marrow cells were induced to differentiate into DCs using GM-CSF and IL-4,followed by maturation stimulation withTNF-α.Exosomes were extracted using ultracentrifugation.Western blot and Amnis image flow cytometry were used to identify exosomes derived from mouse DCs.Amnis image flow cytometry was used to detect the expression of immune-related proteins CD80,CD11c,MHC-Ⅰ,and MHC-Ⅱ in mouse DCs and their exosomes.Results After 5 days of in vitro cultivation,more than 50%of dendritic cells expressed CD80,CD11c,MHC-Ⅰ,and MHC-Ⅱ,reaching the highest level on day 13.The positivity rates were as follows:CD80(97.29±0.63)%,CD11c(92.31±1.18)%,MHC-Ⅰ(97.91±0.49)%,and MHC-Ⅱ(97.91±0.49)%.The differences were statistically significant(P<0.001).The expression gradually decreased after day 13,but approximately 80%of DC cells still expressed MHC-Ⅰ and MHC-Ⅱ immune molecules on day 30.The expression levels of CD80,CD11c,and MHC-Ⅱ on the exosome membrane were highest on day 5 and then decreased overall with prolonged cultivation time,except for MHC-Ⅰ molecules.The differences were statistically significant(P<0.01).Conclusions In vitro-cultured mouse dendritic cells express high levels of immune-related membrane proteins and can be stably maintained for a long time under suitable culture conditions.The secreted exosomes also carry abundant immune-related membrane proteins,but no significant correlation was found between the immune-related proteins on the dendritic cell surface and the exosome membrane surface.
5.Study on the effect of addition or subtraction of Astragalus membranaceus and Draba nemorosa on CHF and changes of cardiac color ultrasound parameters
Lepin GE ; Meise LIN ; Zuchao JIANG ; Pengpeng CHEN ; Longmeng CAO ; Yan LI
China Modern Doctor 2024;62(12):67-72
Objective To analyze the efficacy of Astragalus membranaceus and Draba nemorosa in the treatment of chronic heart failure(CHF),the change of cardiac color ultrasound parameters and its predictive value for adverse end events.Methods A total of 92 patients with Qi-deficiency and blood-stasis combined with Shuiyin CHF treated in Wenzhou Hospital of Traditional Chinese Medicine from September 2019 to September 2021 were randomly selected and divided into two groups by random number table method,with 46 cases in each group.Patients in control group were received conventional Western medicine treatment,and patients in observation group were received the addition and subtraction treatment of Yiqi Huoxue Lishui prescription based on Astragalus membranaceus and Draba nemorosa.The clinical efficacy,TCM syndrome score,heart color ultrasound parameters and the incidence of adverse endpoint events within 6 months were compared between the two groups.Cardiac color doppler ultrasound parameters were compared between the occurrence group and the non-occurrence group,and receiver operating characteristic curve was drawn to analyze the predictive value of cardiac color Doppler ultrasound parameters for adverse end events.Results The total clinical effective rate of the observation group(95.65%)was higher than that of the control group(71.74%)(P<0.05).After treatment,palpitation,weakness,shortness of breath,lethargy and lip purple score in observation group were lower than those in control group(P<0.05).The left ventricular end-diastolic diameter(LVEDD)and left arial diameters(LAD)of the observation group were lower than those of the control group after treatment(P<0.05).After treatment,the left ventricular ejection fraction(LVEF)of the observation group was higher than that of the control group(P<0.05);The incidence of adverse endpoint events within 6 months in the observation group(2.17%)was lower than that in the control group(19.57%)(P<0.05).The LVEDD and LAD in the adverse end point event group were higher than those in the without adverse end point event group(P<0.05),and the LVEF was lower than that in the without adverse end point event group(P<0.05).The area under the curve(AUC)of LVEDD,LAD and LVEF combined to predict the occurrence of adverse endpoint events was 0.812,and the 95%CI confidence interval was 0.734-0.968;The sensitivity of combined detection(93.42%)was higher than that of single detection(71.52%,75.11%,79.62%)(P<0.05).The specificity of combined detection(82.27%)compared with that of single detection(70.19%,73.07%,77.28%)(P>0.05).Conclusion Supplementing qi and promoting blood circulation and rehydrating water based on Astragalus membranaceus and Draba nemorosa can effectively improve the cardiac function of CHF patients,alleviate symptoms such as palpitations and fatigue,and reduce the incidence of adverse endpoint events.In addition,the combined detection of cardiac ultrasound parameters can improve the prediction efficiency of adverse endpoint events,which has certain clinical value.
6.National bloodstream infection bacterial resistance surveillance report (2022) : Gram-negative bacteria
Zhiying LIU ; Yunbo CHEN ; Jinru JI ; Chaoqun YING ; Qing YANG ; Haishen KONG ; Haifeng MAO ; Hui DING ; Pengpeng TIAN ; Jiangqin SONG ; Yongyun LIU ; Jiliang WANG ; Yan JIN ; Yuanyuan DAI ; Yizheng ZHOU ; Yan GENG ; Fenghong CHEN ; Lu WANG ; Yanyan LI ; Dan LIU ; Peng ZHANG ; Junmin CAO ; Xiaoyan LI ; Dijing SONG ; Xinhua QIANG ; Yanhong LI ; Qiuying ZHANG ; Guolin LIAO ; Ying HUANG ; Baohua ZHANG ; Liang GUO ; Aiyun LI ; Haiquan KANG ; Donghong HUANG ; Sijin MAN ; Zhuo LI ; Youdong YIN ; Kunpeng LIANG ; Haixin DONG ; Donghua LIU ; Hongyun XU ; Yinqiao DONG ; Rong XU ; Lin ZHENG ; Shuyan HU ; Jian LI ; Qiang LIU ; Liang LUAN ; Jilu SHEN ; Lixia ZHANG ; Bo QUAN ; Xiaoping YAN ; Xiaoyan QI ; Dengyan QIAO ; Weiping LIU ; Xiusan XIA ; Ling MENG ; Jinhua LIANG ; Ping SHEN ; Yonghong XIAO
Chinese Journal of Clinical Infectious Diseases 2024;17(1):42-57
Objective:To report the results of national surveillance on the distribution and antimicrobial resistance profile of clinical Gram-negative bacteria isolates from bloodstream infections in China in 2022.Methods:The clinical isolates of Gram-negative bacteria from blood cultures in member hospitals of national bloodstream infection Bacterial Resistant Investigation Collaborative System(BRICS)were collected during January 2022 to December 2022. Antibiotic susceptibility tests were conducted by agar dilution or broth dilution methods recommended by Clinical and Laboratory Standards Institute(CLSI). WHONET 5.6 and SPSS 25.0 software were used to analyze the data.Results:During the study period,9 035 strains of Gram-negative bacteria were collected from 51 hospitals,of which 7 895(87.4%)were Enterobacteriaceae and 1 140(12.6%)were non-fermenting bacteria. The top 5 bacterial species were Escherichia coli( n=4 510,49.9%), Klebsiella pneumoniae( n=2 340,25.9%), Pseudomonas aeruginosa( n=534,5.9%), Acinetobacter baumannii complex( n=405,4.5%)and Enterobacter cloacae( n=327,3.6%). The ESBLs-producing rates in Escherichia coli, Klebsiella pneumoniae and Proteus spp. were 47.1%(2 095/4 452),21.0%(427/2 033)and 41.1%(58/141),respectively. The prevalence of carbapenem-resistant Escherichia coli(CREC)and carbapenem-resistant Klebsiella pneumoniae(CRKP)were 1.3%(58/4 510)and 13.1%(307/2 340);62.1%(36/58)and 9.8%(30/307)of CREC and CRKP were resistant to ceftazidime/avibactam combination,respectively. The prevalence of carbapenem-resistant Acinetobacter baumannii(CRAB)complex was 59.5%(241/405),while less than 5% of Acinetobacter baumannii complex was resistant to tigecycline and polymyxin B. The prevalence of carbapenem-resistant Pseudomonas aeruginosa(CRPA)was 18.4%(98/534). There were differences in the composition ratio of Gram-negative bacteria in bloodstream infections and the prevalence of main Gram-negative bacteria resistance among different regions,with statistically significant differences in the prevalence of CRKP and CRPA( χ2=20.489 and 20.252, P<0.001). The prevalence of CREC,CRKP,CRPA,CRAB,ESBLs-producing Escherichia coli and Klebsiella pneumoniae were higher in provinicial hospitals than those in municipal hospitals( χ2=11.953,81.183,10.404,5.915,12.415 and 6.459, P<0.01 or <0.05),while the prevalence of CRPA was higher in economically developed regions(per capita GDP ≥ 92 059 Yuan)than that in economically less-developed regions(per capita GDP <92 059 Yuan)( χ2=6.240, P=0.012). Conclusions:The proportion of Gram-negative bacteria in bloodstream infections shows an increasing trend,and Escherichia coli is ranked in the top,while the trend of CRKP decreases continuously with time. Decreasing trends are noted in ESBLs-producing Escherichia coli and Klebsiella pneumoniae. Low prevalence of carbapenem resistance in Escherichia coli and high prevalence in CRAB complex have been observed. The composition ratio and antibacterial spectrum of bloodstream infections in different regions of China are slightly different,and the proportion of main drug resistant bacteria in provincial hospitals is higher than those in municipal hospitals.
7.A multi-dimensional analysis of pollen broadcasting concerns in Chinese population: a large-scale multi-center cross-sectional survey
Chiyu XU ; Yanshu ZHANG ; Ning LUAN ; Xiangyi LIU ; Dayang QIN ; Hongmin WANG ; Xuping XIAO ; Shuihong ZHOU ; Jie ZHANG ; Ping ZHANG ; Yuqing BAI ; Pengpeng WANG ; Yan QI ; Zhongwu SUN ; Zhuang LIU ; Luo BA ; Wenchao WANG ; Xing LU ; Min WANG ; Rui GUO ; Deyi SUN ; Liyuan TAO ; Li ZHU
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2024;59(1):2-11
Objective:To investigate the concern about pollen broadcasting in Chinese population from multiple dimensions and to understand the information about allergic rhinitis (AR) in China by analyzing related factors.Methods:From March 1 to September 30, 2022, a large-scale multi-center cross-sectional survey was conducted based on the Questionnaire Star platform in 21 Chinese hospitals. A total of 7 056 subjects from 7 regions in China: Northeast, North, East, Central, South, Southwest, and Northwest China were included. Basic characteristics (including social demographic characteristics and disease characteristics of AR patients), concern about pollen broadcasting, the willingness of pollen-induced AR (PiAR) patients to receive pollen broadcasting, and the treatment satisfaction rate of AR patients were collected. The chi-square test, multivariate linear regression model, and Logistic regression analysis were used to analyze the concern about pollen broadcasting in the Chinese population and related factors from multiple dimensions.Results:Among 7 056 subjects, 23.02% were concerned about pollen broadcasting. Among 3 176 self-reported AR and 1 019 PiAR patients, 25.60% and 39.16% were concerned about pollen broadcasting, respectively, which was higher than that of non-AR or non-PiAR subjects ( χ2 value was 21.74 and 175.11, respectively, both P<0.001). Among AR patients, the proportion of spring and autumn allergen-positive patients concerned about pollen broadcasting was higher than that in perennial allergen-positive patients ( χ2 value was 20.90 and 19.51, respectively, both P<0.001). The proportion of AR patients with asthma, sinusitis, allergic conjunctivitis, and cardiovascular and cerebrovascular diseases was higher than those without complications ( χ2 value was 50.83, 21.97, 56.78, 7.62, respectively, all P<0.05). The proportion of AR patients in North China who could find pollen broadcasting locally was 31.01%, significantly higher than those in other regions (all P<0.05). Multivariate linear regression model analysis showed that among PiAR patients, those with higher per capita household income and higher AR disease cognition levels had been concerned about pollen broadcasting in the past, and those complicated with allergic conjunctivitis had stronger intention to receive pollen broadcasting (B value was 0.24, 0.13, 0.66, 0.47, respectively, all P<0.05). The higher the disease cognition level of PiAR patients, the stronger their willingness to actively participate in treatment ( R2=0.72, P<0.001). Only 18.89% of AR patients felt satisfied with the treatment effect. Logistic regression analysis showed that in AR patients, the treatment satisfaction rate was significantly higher among those concerned about pollen broadcasting compared to those who were not ( OR=1.83, P<0.001). Conclusions:Currently, the dissemination of pollen broadcasting in China is hindered by various factors such as disease cognition level. The treatment satisfaction among AR patients remains unsatisfactory.
8.Exploring Pharmacodynamic Material Basis and Mechanism of Hei Xiaoyaosan in Improving Learning and Memory Ability of Insomnia Rats Based on UPLC-Q-TOF-MS and Network Pharmacology
Jiamin LIU ; Hai HUANG ; Pengpeng LIANG ; Yale WANG ; Guiyun LI ; Hongyan WU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(21):19-30
ObjectiveBased on ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS), network pharmacology and pharmacodynamics, to investigate the pharmacodynamic material basis and mechanism of Hei Xiaoyaosan in improving learning and memory ability of insomnia rats. MethodUPLC-Q-TOF-MS was used to characterize the chemical constituents of Hei Xiaoyaosan. Network pharmacology was applied to construct the network of active ingredients-intersecting targets-pathways, and molecular docking was performed on key ingredients and core targets. Sixty 8-week-old male SD rats were selected and randomly divided into blank group, model group, Hei Xiaoyaosan low, medium, and high dose groups(3.82, 7.65, 15.30 g·kg-1), and zolpidem tartrate group(0.5 mg·kg-1), with 10 rats in each group. Except for the blank group, the insomnia model was induced by intraperitoneal injection of p-chlorophenylalanine(PCPA) for 4 consecutive days. Rats in each dosing group were administered the corresponding dose by gavage, once a day for 14 consecutive days. Morris water maze test was utilized to assess the learning and memory ability of rats, transmission electron microscopy was employed to examine the ultrastructure of hippocampal synapses, enzyme-linked immunosorbent assay(ELISA) was conducted to analyze the levels of 5-hydroxytryptamine(5-HT), interleukin-6(IL-6), and tumor necrosis factor-α(TNF-α) in hippocampal tissues, and Western blot was performed to detect the expression levels of tumor suppressor protein p53(TP53), rat sarcoma virus(RAS), epidermal growth factor receptor(EGFR), cyclic adenosine monophosphate(cAMP)-response element binding protein(CREB) binding protein(CREBBP), glycogen synthase kinase-3β(GSK-3β), protein kinase B1(Akt1), nitric oxide synthase 1(NOS1), phosphorylated(p)-Akt1, and p-GSK-3β in hippocampal tissues. Additionally, real-time fluorescence quantitative polymerase chain reaction(Real-time PCR) was used to assess the mRNA expression levels of TP53, RAS, EGFR, CREBBP, GSK-3β, Akt1 and NOS1. ResultA total of 176 components were identified in Hei Xiaoyaosan, mainly flavonoids, triterpene saponins, phenylpropanoids and other compounds. Network pharmacological analysis revealed that TP53, V-Ha-Ras Harvey Rat sarcoma viral oncogene homolog(HRAS), neuroblastoma sarcoma viral oncogene homolog(NRAS), EGFR, CREBBP, GSK-3β, Akt1 and NOS1 were the key targets of Hei Xiaoyaosan in treating insomnia. The core targets were predominantly associated with cAMP, RAS, Ras-associated protein 1(Rap1), advanced glycation end products(AGE)/receptor for AGE(RAGE), and EGFR signaling pathways, and the key active ingredients of Hei Xiaoyaosan in treating insomnia were 8-shogaol, ligustilide F, 6-gingerol, levistilide A and senkyunolide E. Animal experiment results demonstrated that Hei Xiaoyaosan medium and high dose groups significantly increased body weight, shortened sleep latency and prolonged sleep duration in insomnia rats(P<0.01), significantly decreased escape latency and increased platform crossing frequency(P<0.01), and improved the pathological changes of hippocampal synaptic ultrastructure. Meanwhile, the two groups could significantly elevate 5-HT level, Akt1 mRNA expression, Akt1 and p-Akt1 protein expression(P<0.01), reduce inflammatory factor levels(P<0.01), and down-regulate protein expression levels of TP53, RAS, NOS1, EGFR, CREBBP, GSK-3β and p-GSK-3β(P<0.01), as well as mRNA expression levels of TP53, RAS, NOS1, EGFR, CREBBP and GSK-3β in hippocampal tissues(P<0.01). ConclusionThis study determined that the five key active ingredients(8-shogaol, ligustilide F, 6-gingerol, levistilide A and senkyunolide E) in Hei Xiaoyaosan may improve the learning and memory ability of insomnia rats by regulating signaling pathways such as cAMP, RAS, and EGFR, providing an important reference for its mechanism research and clinical application.
9.Analysis of the factors influencing the mental health status of anesthesiology residents during standardized training
Tao TAO ; Yan LIU ; Yuan LI ; Yang ZHANG ; Xueqin NIU ; Pengpeng WANG
Chinese Journal of Medical Education Research 2024;23(11):1572-1578
Objective:To assess the mental health status of anesthesiology residents and identify its influencing factors during standardized training, and to enhance the attention and understanding of standardized training management personnel regarding the mental health of residents.Methods:All on-duty residents in the anesthesiology departments of ten standardized anesthesiology training bases, including The Second Affiliated Hospital of Harbin Medical University, were selected as subjects. Self-designed questionnaires were distributed, which included a general information survey, the Self-Rating Anxiety Scale (SAS), the Self-Rating Depression Scale (SDS), the Fatigue Severity Scale (FSS), and the Self-Rating Scale of Sleep (SRSS), to investigate the psychological state and sources of stress. Based on the questionnaire results, residents from the Department of Anesthesiology at The Second Affiliated Hospital of Harbin Medical University were purposively and heterogeneously selected as interviewees. One-on-one semi-structured in-depth interviews were conducted to understand the major life events that triggered emotional or psychological issues and their connections with standardized training or the profession. This survey also aimed to understand the support that anesthesiology residents hoped to receive and their suggestions for the standardized training program. The chi-square test, t-test, and one-way analysis of variance were performed using Graphpad Prism 9.0. Interview content was coded and categorized using data analysis software NVivo12. The Colaizzi's seven-step method was used to analyze the data and extract interview themes. Results:A total of 157 valid questionnaires were recovered, with a response rate of 100.00%. The mean scores of SAS, SDS, FSS, and SRSS were (50.60±14.16), (56.31±12.90), (44.49±13.88), and (27.78±9.40), respectively, all meeting the diagnostic criteria for mild conditions. Of them, 16 (10.19%), 18 (11.46%), 47 (29.94%), and 19 (12.10%) residents met the diagnostic criteria for severe conditions. Compared with Grade 1 residents, Grade 3 residents showed increased scores in anxiety [(54.24±13.83) vs. (47.02±12.04), P=0.030] and depression [(59.66±12.74) vs. (51.53±10.76), P=0.005]. In total, 24 anesthesiology residents were selected for semi-structured interview and three themes were identified: psychological stress related to the training bases, psychological stress related to personal issues, and psychological stress related to career prospects. Conclusions:The anesthesiology residents during standardized training have high prevalence of psychological disorders, which are closely related to work overload, financial stress, occupational exposure, and personal factors. Psychological counseling and crisis intervention should be regarded as integral components of standardized training management, and attention is needed in monitoring the psychological fluctuations of anesthesiology residents and helping them to complete their rotations on time.
10.Clinical features analysis of 65 cases of multiple myeloma with extramedullary disease
Jichun SHEN ; Yuchen ZHANG ; Yang LI ; Qi DENG ; Pengpeng XU
Clinical Medicine of China 2024;40(3):206-211
Objective:To study the clinical features, treatment and prognosis of multiple myeloma (MM)complicated with extramedullary disease (EMD).Methods:A total of 65 patiens admitted to the Characteristic Medical Center and Tianjing First Central Hospital from January 2014 to May 2020 were analyzed retrospectively as the study subjects. The age ranged from 22 to 79 years, with a median age of 56(22,79) years, and the ratio of female to male was 27/38. They were respectively treated by chemotherapy, radiotherapy or autologous hematopoietic stem cell transplantation (AHSCT). Fluorescence in situ hybridization (FISH) was used in all 65 patients to detect t(4;14), t(14;16), 1q21, del(17p). The measurement data of non-normal distribution were expressed in M (Q1,Q3), and the survival curve was constructed by Kaplan-Meier method.Result:The increase of β2- microglobulin and lactate dehydrogenase, multiple osteolytic lesions were found in all patients. The most common rate of chromosomal abnormalities were10.8% for t(4;14), 16.9% for t(14;16), 18.5% for 1q21, 21.5% for del(17p). Both 1q21 and del(17p) were co-existed in 7.7% of patients. The effective rate of 65 patients treated with V-DRPACE regimen was 89.2%(58/65). Thirty-two patients who were in partial remission were treated with radiotherapy. The complete remission rate of radiotherapy was 18.8%(6/32). Twenty patients eligible for transplantation underwent AHSCT and 13 patients achieved complete remission. At the end of follow-up, the median overall survival(OS) of 65 patients was 44.8 months, and progression-free survival (PFS) was11 months.Conclusion:MM with EMD usually involve multiple sites, prone to multiple osteolytic lesions, increased β2- microglobulin and lactate dehydrogenase. The rate of cytogenetic abnormalities in such patients is significantly increased. The V-DRPACE regimen is effective in treating MM patients with extramedullary lesions and can benefit from AHSCT.

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