1.Dipsacus asper Treats Alzheimer's Disease in Caenorhabditis elegans by Regulating PPARα/TFEB Pathway
Mengmeng WANG ; Jianping ZHAO ; Limin WU ; Shuang CHU ; Yanli HUANG ; Zhenghao CUI ; Yiran SUN ; Pan WANG ; Hui WANG ; Zhenqiang ZHANG ; Zhishen XIE
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(5):104-114
ObjectiveTo investigate the anti-Alzheimer's disease (AD) effect of Dipsacus asper(DA) in the Caenorhabditis elegans model, and decipher the underlying mechanism via the peroxisome proliferator-activated receptor α (PPARα)/transcription factor EB (TFEB) pathway. MethodsFirst, transgenic AD C. elegans individuals were assigned into the blank control, model, positive control (WY14643, 20 µmol·L-1), and low-, medium-, and high-dose (100, 200, and 400 mg·L-1, respectively) DA groups. The amyloid β-42 (Aβ42) formation in the muscle cells, the paralysis time, and the deposition of amyloid β-protein (Aβ) in the head were detected. The lysosomal autophagy in the BV2 cell model was examined by Rluc-LC3wt/G120A. The expression levels of lysosomal autophagy-related proteins LC3Ⅱ, LC3I, LAMP2, and TFEB were detected by Western blot. Real-time quantitative polymerase chain reaction (Real-time PCR) was employed to determine the mRNA levels of autophagy-related genes beclin1 and Atg5 and lysosome-related genes LAMP2 and CLN2 downstream of PPARα/TFEB. A reporter gene assay was used to detect the transcriptional activities of PPARα and TFEB. Immunofluorescence was used to detect the fluorescence intensity of PPARα, and the active components of the ethanol extract of DA were identified by UPLC-MS. RCSB PDB, Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP), and Autodock were used to analyze the binding between the active components and PPARα-ligand-binding domain (LBD). ResultsCompared with the model group, the positive control group and 200 and 400 mg·L-1 DA groups showed prolonged paralysis time (P<0.05), and all the treatment groups showed decreased Aβ deposition in the head (P<0.01). DA within the concentration range of 50-500 mg·L-1 did not affect the viability of BV2 cells. In addition, DA enhanced the autophagy flux (P<0.05), up-regulated the mRNA levels of beclin1, Atg5, LAMP2, and CLN2 (P<0.05, P<0.01), promoted the nuclear translocation of TFEB (P<0.05), increased LAMP2 expression and autophagy flux (P<0.05, P<0.01), and enhanced the transcriptional activities of PPARα and TFEB (P<0.01). The positive control group and 200 and 400 mg·L-1 DA groups showed enhanced fluorescence intensity of PPARα in the BV2 nucleus (P<0.01). UPLC-MS detected nine known compounds of DA, from which 8 active components of DA were screened out. The docking results suggested that a variety of components in DA could bind to PPARα-LBD and form stable hydrogen bonds. ConclusionDA may reduce the pathological changes in AD by regulating the PPARα-TFEB pathway.
2.Low disease activity and remission status of systemic lupus erythematosus in a real-world study
Limin REN ; Chuchu ZHAO ; Yi ZHAO ; Huiqiong ZHOU ; Liyun ZHANG ; Youlian WANG ; Lingxun SHEN ; Wenqiang FAN ; Yang LI ; Xiaomei LI ; Jibo WANG ; Yongjing CHENG ; Jiajing PENG ; Xiaozhen ZHAO ; Miao SHAO ; Ru LI
Journal of Peking University(Health Sciences) 2024;56(2):273-278
Objective:To investigate the rates of low disease activity and clinical remission in patients with systemic lupus erythematosus(SLE)in a real-world setting,and to analyze the related factors of low disease activity and clinical remission.Methods:One thousand patients with SLE were enrolled from 11 teaching hospitals.Demographic,clinical and laboratory data,as well as treatment regimes were collec-ted by self-completed questionnaire.The rates of low disease activity and remission were calculated based on the lupus low disease activity state(LLDAS)and definitions of remission in SLE(DORIS).Charac-teristics of patients with LLDAS and DORIS were analyzed.Multivariate Logistic regression analysis was used to evaluate the related factors of LLDAS and DORIS remission.Results:20.7%of patients met the criteria of LLDAS,while 10.4%of patients achieved remission defined by DORIS.Patients who met LLDAS or DORIS remission had significantly higher proportion of patients with high income and longer disease duration,compared with non-remission group.Moreover,the rates of anemia,creatinine eleva-tion,increased erythrocyte sedimentation rate(ESR)and hypoalbuminemia was significantly lower in the LLDAS or DORIS group than in the non-remission group.Patients who received hydroxychloroquine for more than 12 months or immunosuppressant therapy for no less than 6 months earned higher rates of LLDAS and DORIS remission.The results of Logistic regression analysis showed that increased ESR,positive anti-dsDNA antibodies,low level of complement(C3 and C4),proteinuria,low household in-come were negatively related with LLDAS and DORIS remission.However,hydroxychloroquine usage for longer than 12 months were positively related with LLDAS and DORIS remission.Conclusion:LLDAS and DORIS remission of SLE patients remain to be improved.Treatment-to-target strategy and standar-dized application of hydroxychloroquine and immunosuppressants in SLE are recommended.
3.Discussion on the Prevention and Treatment of Urolithiasis by Integrated Traditional Chinese and Western Medicine based on “Shen-kidney Theory”
Bing SHI ; Fan ZHAO ; Lyuzhong XIE ; Chunhe ZHANG ; Bodong LYU ; Limin MA ; Guozheng QIN
Journal of Traditional Chinese Medicine 2024;65(6):572-576
The pathogenesis of urolithiasis is not yet clear, and there are obvious limitations in the prevention and treatment of urolithiasis by either Chinese or western medicine. The microscopic pathological changes of the kidney from anatomical perspective have a certain internal connection with viewpoint of “kidney storing insufficiency, and kidney deficiency as the root” in the traditional Chinese medicine (TCM) zang-fu (脏腑) theory. Accordingly, the prevention and treatment of urolithiasis by integrated traditional Chinese and western medicine based on “shen-kidney” theory has been proposed. It is believed that the prevention and treatment of urolithiasis can be divided into two stages, that is expelling stones and preventing stones. In terms of preventing stones from kideny, it is recommended to focus on the early pathological changes of the kidney; for preventing stones from shen, it is advised to prevent and treat urolithiasis from kidney deficiency. The treatment should be time-based and stage by stage. Adhering to the principle of “prevention before disease occurs, prevention is more important than treatment” aims to advance the intervention targets for the prevention and treatment of urolithiasis. Emphasizing on the simultaneous treatment of kidney disease and urolithiasis, it is critical to put focus on the development of calcium-containing crystalline nephropathy in the early stage of stone formation, as well as the fundamental pathogenesis of kidney deficiency in TCM. Shen-kidney theory aims to further promote the integration of traditional Chinese and western medicine in the prevention and treatment of urolithiasis, which may provide certain reference for solving the current dilemma of urolithiasis prevention and treatment.
4.Traditional Chinese Medicine Regulates Oxidative Stress-related Signaling Pathways to Prevent and Treat Bronchial Asthma: A Review
Xuyang ZHENG ; Zhenhui SHU ; Yi LI ; Ziyue QU ; Weilin LI ; Kexin MA ; Junsheng SHA ; Limin ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(18):260-269
Bronchial asthma (asthma for short) is a common clinical respiratory disease mainly characterized by chronic airway inflammation, with complicated pathogenesis and a long treatment cycle. It is lingering and difficult to be cured, and lack specific drugs. Oxidative stress is a new focus in the research on the pathogenesis of asthma and a potential key target for the treatment. Under physiological conditions, the oxidative and antioxidative systems in the body are in a dynamic balance, and the two antagonize each other to maintain normal life activities. In the case of asthma attack, oxidation products such as reactive oxygen species (ROS), malondialdehyde (MDA), and nitric oxide (NO) are produced excessively, while the content of antioxidants such as superoxide dismutase (SOD), catalase (CAT), and glutathione (GSH) is reduced. As a result, the oxidation exceeds the removal of oxidation products, which aggravates oxidative stress. In addition, the overproduction of ROS activates oxidative stress-related signaling pathways to produce pro-inflammatory factors, exacerbating inflammation, which leads to lung and airway tissue damage. In recent years, traditional Chinese medicine has garnering increasing attention because of the unique advantages in the treatment of asthma, especially in regulating redox balance, alleviating oxidative stress in asthma patients, and reducing inflammation. On the one hand, by inhibiting the mitogen-activated protein kinase (MAPK) and transcription factor (NF)-κB signaling pathways, traditional Chinese medicine can reduce the content of oxidation products and pro-inflammatory factors from the source. On the other hand, by activating the nuclear factor-erythroid 2-related factor 2 (NrF2) signaling pathway, traditional Chinese medicine can elevate the levels of antioxidant enzymes and enhance the antioxidant system to neutralize the excessive accumulation of oxidation products. Therefore, the adjustment of redox balance state by traditional Chinese medicine may be a new means and a new direction for the prevention and treatment of asthma in the future. This paper summarizes the oxidative stress-related pathways in the pathogenesis of asthma and reviews the latest research progress in the regulation of oxidative stress-related pathways by Chinese medicine extracts and prescriptions in the treatment of asthma, with a view to providing a fuller, more solid, and more scientific theoretical basis for the clinical and basic research on the prevention and treatment of asthma by traditional Chinese medicine.
5.Distribution of Nardosinone and Its Metabolites in Rats Analyzed by UPLC-Q-Exactive Orbitrap MS/MS
Lijuan SHI ; Xuelian ZHAO ; Yifan TIAN ; Limin LI ; Yuan LIU ; Xiaojiang ZHOU ; Yanfei HUANG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(22):187-195
ObjectiveUltra-performance liquid chromatography-quadrupole/electrostatic field orbitrap high resolution mass spectrometry(UPLC-Q-Exactive Orbitrap MS/MS) was used to investigate the metabolism and distribution of nardosinone in rats, then metabolic pathways were speculated. MethodRats were administered with 30 mg·kg-1 of nardosinone suspension by gavage for 3 consecutive days, and plasma, urine, feces, and tissues of heart, liver, spleen, lung, kidney, brain, stomach, and intestine were collected at predetermined time points. After treatment, the samples were processed for UPLC-Q-Exactive Orbitrap MS/MS, and the MS data were analyzed using Xcalibur 2.2 software. The metabolites were searched by comparing the base peak chromatogram and extracted ion chromatogram between the treated group and blank group, and based on the relative retention time(tR), quasi-molecular ion peak, precise molecular mass, and fragment ions of MS/MS, the elemental composition were searched using databases such as SciFinder and PubChem, as well as referring to relevant literature, the possible metabolites were identified and the metabolic pathways were inferred. ResultA total of 30 metabolites of nardosinone were identified, including 15, 19, 12, 7, 4, 11, 8, 13, 13, 8 and 12 metabolites in urine, feces, plasma, brain, heart, liver, spleen, lung, kidney, stomach and intestine, respectively. The main metabolic pathways of nardosinone in rats were hydroxylation, dehydroxylation, reduction, dehydrogenation, hydration, dehydration, carboxylation, glucuronidation, and dehydroxy-isopropyl. ConclusionNardosinone can be metabolized by phase Ⅰ and phase Ⅱ metabolism in rats, and the metabolites are widely distributed in the major organs. The results of this study can provide a basis for further research on the pharmacodynamic material basis, pharmacological mechanism and clinical application of nardosinone.
6.Study on the Distribution and Clinical Characteristics of TCM Syndromes in Infertility Patients with Autoimmune Thyroiditis
Xiaoru FANG ; Limin WU ; Qunying FANG ; Mengli WANG ; Shiyu ZHAO ; Zonghui QIAO ; Hongbing LUAN ; Zhaohui FANG
Chinese Journal of Information on Traditional Chinese Medicine 2024;31(1):164-170
Objective To investigate the distribution and clinical characteristics of TCM syndromes in infertility patients with autoimmune thyroiditis(AIT).Methods By referring to the relevant literature on the distribution of TCM syndromes of AIT and infertility in women of childbearing age,the TCM Syndromes Survey of Infertility Patients with Autoimmune Thyroiditis was formulated.256 cases of infertility patients with AIT who underwent in vitro fertilization-embryo transfer(IVF-ET)at the Reproductive Center of The First Affiliated Hospital of University of Science and Technology of China from June 2019 to December 2020 were retrospectively analyzed.The data of basic information,TCM syndromes,basic thyroid-stimulating hormone(TSH),thyroid peroxidase antibody(TPO-Ab)and thyroglobulin antibody(TG-Ab)were collected.By means of frequency analysis and systematic cluster analysis,the distribution regularity of TCM syndromes in infertility patients with AIT were concluded.Results After analysis,the following 5 common syndrome elements were obtained,namely,qi deficiency,liver depression,spleen deficiency,kidney deficiency and blood stasis.In addition,it was concluded that this disease was more common in complex syndromes.Through systematic clustering analysis,main TCM syndromes of this disease were obtained,which were qi deficiency and liver depression syndrome(69 cases,26.9%),spleen and kidney deficiency syndrome(45 cases,17.6%),spleen qi deficiency syndrome(38 cases,14.8%),qi deficiency and blood stasis syndrome(36 cases,14.1%),kidney deficiency and liver depression syndrome(32 cases,12.5%)and other syndrome types(36 cases,14.1%).The basic TSH level was higher in patients with qi deficiency and liver depression syndrome,spleen qi deficiency syndrome,qi deficiency and blood stasis syndrome than other syndrome types,with statistical significance(P<0.05).There was no significant difference in TPO-Ab and TG-Ab titers among different syndromes(P>0.05).Conclusion TCM syndromes of infertility patients with AIT can be clustered into qi deficiency and liver depression syndrome,spleen and kidney deficiency syndrome,spleen qi deficiency syndrome,qi deficiency and blood stasis syndrome,kidney deficiency and liver depression syndrome and other syndrome.The main element of syndrome is qi deficiency,and the pathological sites involved spleen,kidney and liver.Stasis blood is a main pathological product.It is required to pay close attention to the thyroid function in AIT patients with qi deficiency.
7.Clinical guidelines for the treatment of ankylosing spondylitis combined with lower cervical fracture in adults (version 2024)
Qingde WANG ; Yuan HE ; Bohua CHEN ; Tongwei CHU ; Jinpeng DU ; Jian DONG ; Haoyu FENG ; Shunwu FAN ; Shiqing FENG ; Yanzheng GAO ; Zhong GUAN ; Hua GUO ; Yong HAI ; Lijun HE ; Dianming JIANG ; Jianyuan JIANG ; Bin LIN ; Bin LIU ; Baoge LIU ; Chunde LI ; Fang LI ; Feng LI ; Guohua LYU ; Li LI ; Qi LIAO ; Weishi LI ; Xiaoguang LIU ; Hongjian LIU ; Yong LIU ; Zhongjun LIU ; Shibao LU ; Yong QIU ; Limin RONG ; Yong SHEN ; Huiyong SHEN ; Jun SHU ; Yueming SONG ; Tiansheng SUN ; Yan WANG ; Zhe WANG ; Zheng WANG ; Hong XIA ; Guoyong YIN ; Jinglong YAN ; Wen YUAN ; Zhaoming YE ; Jie ZHAO ; Jianguo ZHANG ; Yue ZHU ; Yingjie ZHOU ; Zhongmin ZHANG ; Wei MEI ; Dingjun HAO ; Baorong HE
Chinese Journal of Trauma 2024;40(2):97-106
Ankylosing spondylitis (AS) combined with lower cervical fracture is often categorized into unstable fracture, with a high incidence of neurological injury and a high rate of disability and morbidity. As factors such as shoulder occlusion may affect the accuracy of X-ray imaging diagnosis, it is often easily misdiagnosed at the primary diagnosis. Non-operative treatment has complications such as bone nonunion and the possibility of secondary neurological damage, while the timing, access and choice of surgical treatment are still controversial. Currently, there are no clinical practice guidelines for the treatment of AS combined with lower cervical fracture with or without dislocation. To this end, the Spinal Trauma Group of Orthopedics Branch of Chinese Medical Doctor Association organized experts to formulate Clinical guidelines for the treatment of ankylosing spondylitis combined with lower cervical fracture in adults ( version 2024) in accordance with the principles of evidence-based medicine, scientificity and practicality, in which 11 recommendations were put forward in terms of the diagnosis, imaging evaluation, typing and treatment, etc, to provide guidance for the diagnosis and treatment of AS combined with lower cervical fracture.
8.Relationship between serum levels of TGF- β1, VEGF and tumor markers and local recurrence after breast-conserving surgery in patients with breast cancer
Shoushuai LI ; Ge ZHAO ; Limin TIAN ; Haijun ZHU
Chinese Journal of Endocrine Surgery 2024;18(5):619-623
Objective:To explore the relationship between serum levels of transforming growth factor- β1 (TGF- β1), vascular endothelial growth factor (VEGF) and tumor markers and local recurrence after breast-conserving surgery in patients with breast cancer. Methods:104 patients with breast cancer who underwent breast-conserving surgery in Xi’ an Central Hospital from Jan. 2020 to Dec. 2022 were selected and followed up for 1 year after surgery. According to the occurrence of local recurrence, they were divided into recurrence group ( n=16) and non-recurrence group ( n=88). The levels of serum TGF- β1, VEGF and tumor markers [carbohydrate antigen 125 (CA125), carcinoembryonic antigen (CEA) ] were compared between the two groups of patients. Univariate and multivariate Logistic regression analyses were used to analyze the influencing factors of local recurrence after breast-conserving surgery. Receiver operating characteristic (ROC) curve was applied to analyze the predictive value of TGF- β1, VEGF, CA125 and CEA on local recurrence after breast-conserving surgery. Results:The levels of VEGF, TGF- β1, CAl25 and CEA in recurrence group [ (358.83±38.00) ng/L, (849.90±74.19) U/mL, (18.34±1.61) ng/L and (40.20±5.64) ng/mL] were higher than those of (296.05±39.57) ng/L (742.85±79.96) ng/L, (14.97±1.66) U/mL and (32.79±4.72) ng/mL in non-recurrence group (all P<0.05). Lymph node metastasis, postoperative adjuvant therapy, tumor diameter and TNM staging were independent risk factors of local recurrence after breast-conserving surgery (all P<0.05). The areas under the ROC curves (AUCs) of serum VEGF, TGF- β1, CAl25 and CEA in predicting local recurrence after breast-conserving surgery were 0.847, 0.834, 0.925 and 0.935 respectively. The AUC of CEA was the largest, with sensitivity of 85.23% and specificity of 100% (all P<0.05) . Conclusions:The levels of TGF- β1, VEGF, CAl25 and CEA in patients with local recurrence after breast-conserving surgery are increased, and their levels are more effective in predicting local recurrence after breast-conserving surgery. Local recurrence after breast-conserving surgery is also affected by lymph node metastasis, postoperative adjuvant therapy, tumor diameter and TNM staging.
9.Study on the Characteristic Spectra and Multi indicator Component Screening and Content Determination of BSLYT Substance Standards Based on UHPLC-MS/MS Method
Wei ZHAO ; Yongfan MA ; Zhongyi LIU ; Limin SONG ; Yanhua LIU ; Shoulin ZHANG ; Hongbo CHU
World Science and Technology-Modernization of Traditional Chinese Medicine 2024;26(3):704-720
Objective To determine the chemical constituents in BSLYT by ultra-high performance liquid chromatography-mass spectrometry(UHPLC-MS/MS),and to establish a high-performance liquid chromatographic fingerprint of the material basis of BSLYT and the content determination of its six main constituents,mononoside,loganin,oroxin A,oroxin B,baicalein and astilbin,providing a reference for the quality control.Methods The mass spectrometry data were used to establish the fingerprints.The content of the main components in the BSLYT samples was calculated by using the external method,and the discrepency between different batches of samples were analyzed by combining with chemometric methods.Results A total of 69 compounds were identified by mass spectrometry,and 13 compounds were identified after comparison with high-performance liquid chromatography(HPLC).The similarity of the baseline characterization profiles of the 15 batches of BSLYT substances was above 0.90,and a total of 27 common peaks were identified.Cluster analysis(CA)classified the substance benchmarks into 2 classes,S1,S2,S5,S8,S9,S13,and S15 were clustered into one class,and S3,S4,S6,S7,S10-S12,and S16 were clustered into one class.By combining PCA and OPLS-DA,the chemical components affecting the baseline classification of the substances were screened and attributed to wood butterfly,cornelian cherry and cocos nigra,respectively.The contents of six components were determined by MICS,which ranged from 0.31%-0.51%,0.12%-0.22%,0.09%-0.19%,0.09%-0.24%,0.07%-0.18%,and 0.08%-0.29%for mononoside,loganin,oroxin A,oroxin B,baicalein and astilbin respectively.Conclusion The fingerprint and multi-indicator content determination method established in this paper are accurate and stable,which provide a basis for the quality control of the material benchmark of Kidney and Pharynx Formula and its related preparations.
10.Changes in Drug Resistance and Cross-resistance Phenotype Analysis of Pseudomonas aeruginosa Clinical Isolates from 2016 to 2022 in Kunming
Zan LU ; Hongyan ZHAO ; Chunfu LI ; Daibi LI ; Yang HE ; Limin YIN
Journal of Modern Laboratory Medicine 2024;39(6):189-194
Objective To provide some evidence for early empirical treatment of infections caused by Pseudanonas aeruginosa(PA)and establish a certain foundation for further studies on its resistance mechanisms.Analyzed the resistance rate of PA and the phenotype characteristics for seven years at this hospital.Methods Using Whonet 5.6 software,the drug sensitivity data of PA isolated and reported from clinical specimens at Kunming First People's Hospital from 2016 to 2022 were analyzed annually.Imipenem,meropenem,ceftazidime,and cefepime were the main observation objects,statisticaly analyze the resistance of other antibiotics under different drug resistance combinations of the above-mentioned drugs Results A total of 1 920 strains of PA were detected in 7 years,with polymyxins exhibiting the lowest resistance rate at 1%.The average resistance rates for Amikacin,Gentamicin and Tobramycin were 6.4%,8.6%and 5.1%,respectively.The resistance rates of Quinolones,Piperacillin/Tazobactam,Ceftazidime,and Cefepime were similar,ranging from 10%~20%.Imipenem and Meropenem did not show better sensitivity,with resistance rates of 21.3%and 18.4%,respectively.The resistance rates of major antibiotics have fluctuated over the past 7 years,mostly increasing from 2017 to reaching a peak in 2018 and then falling back.And then stabilized with the domestic average level.1 393 strains were sensitive to Imipenem,and the sensitivity rate of Meropenem in this part was 96.2%.1 451 strains were sensitive to Imipenem,and 89%of them were sensitive to Meropenem at the same time.369 strains were both resistant to Imipenem and Meropenem,and 33.1%of them were sensitive to Ceftazidime.The more resistant to β-lactam,the lower the sensitivity rate to Amikacin and Ciprofloxacin.Conclusion The overall drug sensitivity of PA in this hospital is relatively good.Should adhere to strict management of antibiotic use.Aminoglycosides can be the preferred drugs for empirical treatment,followed by Quinolones,and Carbapenems and Ceftazidime are also candidates.PA has diverse resistance mechanisms to β-lactam antimicrobials,and carbapenem-resistant strains can be mediated by different mechanisms or jointly.

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