1.Treatment of Sepsis-induced Inflammatory Responses with Xijiao Dihuangtang by Modulation of PKM2-mediated One-carbon Metabolism Pathway
Qixiang YAN ; Yeyan ZHU ; Fan GE ; Qimeng SUN ; Leyao YE ; Fang TIAN ; Jun LU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(10):18-26
ObjectiveTo investigate the effects of Xijiao Dihuangtang (XJDHT) on mice with sepsis and cellular models of sepsis and explore its molecular mechanism in alleviating sepsis-induced inflammatory responses via regulating pyruvate kinase M2 (PKM2)-mediated one-carbon metabolism pathway. MethodsForty C57BL/6N mice were randomly divided into four groups: normal group, model group, low-dose XJDHT group (7.7 g·kg-1), and high-dose XJDHT group (15.4 g·kg-1). After one week of continuous gavage, sepsis was induced using cecal ligation and puncture (CLP) in groups except the normal group. 24 h after the surgery, mortality rates in all groups were recorded, and serum cytokines were measured by enzyme linked immunosorbent assay (ELISA). Lung histopathology was examined by hematoxylin-eosin (HE) staining. During the in vitro experiment, the human monocytic leukemia cell line (THP-1) was exposed to various concentrations of XJDHT and treated with lipopolysaccharide (LPS) at a final concentration of 2 mg·L-1 for 24 h. Cell apoptosis was detected using terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. Protein levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), B-cell lymphoma 2 (Bcl-2), and Bcl-2-associated X protein (Bax) were measured by Western blot. Transcriptome sequencing was performed to analyze differentially expressed genes in all groups and conduct gene ontology (GO) enrichment. Key genes in the one-carbon metabolism pathway, including pyruvate kinase M2 (PKM2), 5-methyltetrahydrofolate-homocysteine methyltransferase (MTR), and phosphoglycerate dehydrogenase (PHGDH), were verified by Western blot. A PKM2 inhibition model was established using shikonin for further protein expression analysis. ResultsAnimal experiments showed that compared with the normal group, the model group exhibited significantly elevated body temperature and lung pathology (P<0.01) and increased serum TNF-α and IL-1β levels (P<0.01). High-dose XJDHT reduced body temperature and lung tissue damage (P<0.01) and significantly decreased serum TNF-α and IL-1β levels (P<0.01). Low-dose XJDHT treatment showed no significant temperature change (P<0.01) but reduced serum TNF-α and IL-1β levels (P<0.01). Transcriptome sequencing and Western blot revealed significant differences in the expression of TNF-α, IL-1β, and one-carbon metabolism genes (PKM2, MTR, and PHGDH) (P<0.01). Cell experiments demonstrated that compared to the normal group, the model group showed elevated protein expressions of TNF-α and IL-1β in THP-1 cells (P<0.01), decreased Bcl-2/Bax ratio, and increased apoptosis (P<0.01). Transcriptome sequencing and Western blot revealed significant differences in the expression of TNF-α, IL-1β, and one-carbon metabolism genes (PKM2, MTR, and PHGDH) (P<0.01). Compared to the model group, high-dose XJDHT significantly increased Bax/Bcl-2 ratio and PHGDH protein expression (P<0.01) and effectively reduced cell apoptosis (P<0.01) while down-regulating protein expressions of TNF-α, IL-1β, PKM2, and MTR (P<0.01). Low-dose XJDHT moderately increased Bax/Bcl-2 ratio and PHGDH protein expression (P<0.05), reduced apoptosis (P<0.05), and decreased IL-1β and MTR protein levels (P<0.05, P<0.01), but there were no significant changes in TNF-α and PKM2 expression. After PKM2 inhibition by shikonin in THP-1 cells, the expression of protein related to one-carbon metabolism was detected. Compared with the blank group, the LPS-induced model group showed significantly upregulated PKM2 and MTR protein expression (P<0.01) and downregulated PHGDH expression (P<0.01). Compared with the model group, shikonin treatment significantly reduced PKM2 expression (P<0.05), increased PHGDH expression (P<0.01), and decreased MTR expression (P<0.05). ConclusionXJDHT can inhibit the release of inflammatory factors in sepsis, and its mechanism is related to the intervention of the PKM2-regulated one-carbon metabolism pathway in macrophages.
2.Determination of sotagliflozin in rat plasma by UPLC-MS/MS method
Ge-xi CAO ; Xiao-xu ZHANG ; Yan-ru DENG ; Bin YAN ; Zhan-jun DONG
The Chinese Journal of Clinical Pharmacology 2025;41(2):230-234
Objective To establish a ultra-high performance liquid chromatography-tandem mass spectrometer(UPLC-MS/MS)method for determining the concentration of sotagliflozin in rat plasma and apply it to pharmacokinetic studies in rats.Methods Electrospray negative ion multi-reaction ion detection was used.Chromatographic column:EXT-C18(2.1 mm × 100.0 mm,2.7 μm);column temperature:45 ℃;mobile phase:5 mmol·L-1 ammonium acetate aqueous solution-acetonitrile;flow rate:0.35 mL·min-1;ion pairs:sotagliflozin m/z 483.3→315.1,dapagliflozin m/z 467.4→329.2;injection volume:6 μL,plasma samples were processed using methyl tert-butyl ether liquid-liquid extraction.Six male SD rats were administered a single oral dose of sogliflozin at 40 mg·kg-1,and detected the concentration of sogliflozin in plasma.Pharmacokinetic parameters were calculated using Drug And Statistics(DAS)2.1.1.Results Sotagliflozin showed good linearity within the range of 5-2 000 ng·mL-1,with intra-day and inter-day precision both less than 15%.The recovery rate,matrix effect,and stability were all within the specified range.Pharmacokinetic parameters:Cmax was(3 716.67±568.28)ng·mL-1,tmax was(1.00±0.32)h,t1/2 was(2.28±0.45)h,AUC0-t was(1.70 × 104±2 075.87)ng·mL-1·h.Conclusion This study established a method for determining the concentration of sotagliflozin in rat plasma,which is characterized by high sensitivity,rapid detection,and good repeatability.It is suitable for the determination of sotagliflozin concentration in plasma and pharmacokinetic studies.
3.LncRNA GS1-124K5.4 targeting regulation of PRDX6 on proliferation,migration and Invasion of lung squamous carcinoma cells
Yu-ning HU ; Yan-lei GE ; Ye JIN ; Jun-qing GAN ; Wei-nan YAO ; Ya-nan WU ; Xuan ZHENG ; Zi-qing LIU ; Xin SU ; Guo-gui SUN
Chinese Pharmacological Bulletin 2025;41(8):1531-1541
Aim To investigate the effect of long-chain non-coding RNA(lncRNA)GS1-124K5.4 targeting regulation of PRDX6 on proliferation,migration and in-vasion of lung squamous carcinoma(LUSC)cells and the underlying mechanism.Methods The expression level of lncRNA GS1-124K5.4 in lung cancer tissues and adjacent tissues of 60 patients with LUSC were de-termined by fluorescence in situ hybridization.The ex-pression level of lncRNA GS1-124K5.4 in human nor-mal lung cells and LUSC cells were determined by qRT-PCR.Two kinds of LUSC cells(NCI-H 1703,SK-MES-1)with highest expression level of lncRNA GS1-124K5.4 were selected for subsequent experi-ments.The distribution of lncRNA GS1-124K5.4 in cells was studied by fluorescence in situ hybridization and prokaryotic separation.The effect of knockdown of lncRNA GS1-124K5.4 on proliferation of NCI-H1703 and SK-MES-1 cells was studied by CCK-8 experiment and cell clone formation experiment;the effect of knockdown of lncRNA GS1-124K5.4 on migration of NCI-H1703 and SK-MES-1 cells was studied by cell scratch experiment and Transwell cell migration experi-ment;and the effect of knockdown of lncRNA GS1-124K5.4 on invasion of NCI-H1703 and SK-MES-1 cells was studied by Transwell invasion experiment.The protein to be bound by lncRNA GS1-124K5.4 was detected by RNA pull-down combined with mass spec-trometry and immune-precipitation.The effect of knockdown of lncRNA GS1-124K5.4 targeting PRDX6 on proliferation,migration and invasion of NCI-H1703 and SK-MES-1 cells was studied.Results(1)The fluorescence intensity of lncRNA GS1-124K5.4 in lung squamous cell carcinoma increased compared with that in adjacent tissues(P<0.05),and the expression of lncRNA GS1-124K5.4 was related with lymph node metastasis and clinical stage(P<0.05).(2)The ex-pression level of lncRNA GS1-124K5.4 in NCI-H1703,NCI-H520 and SK-MES-1 cells significantly increased(P<0.05).(3)The result of fluorescence in situ hybridization experiment and nucleoplasm sepa-ration experiment showed that lncRNA GS1-124K5.4 was mainly distributed in cell nucleus.(4)The prolif-eration,migration and invasion ability of NCI-H1703 and SK-MES-1 cells with knockdown of lncRNA GS1-124K5.4 significantly decreased(P<0.05).(5)PRDX6 protein to be bound to LncRNA GS1-124K5.4 was determined by RNA pull-down combined with mass spectrometry and immunoprecipitation.(6)The prolif-eration,migration and invasion ability of NCI-H1703 and SK-MES-1 cells with overexpression of lncRNA GS1-124K5.4 significantly increased(P<0.05);the proliferation,migration and invasion ability of NCI-H1703 and SK-MES-1 cells with knockdown of PRDX6 significantly decreased(P<0.05);the proliferation,migration and invasion ability of NCI-H1703 and SK-MES-1 cells with overexpression of lncRNAGS1-124K5.4 and knockdown of PRDX6 showed no signifi-cant change(P>0.05).Conclusions LncRNA GS1-124K5.4 is highly expressed in lung squamous cell carcinoma,and it may promote the proliferation,migration and invasion of lung squamous carcinoma cells by targeting the expression of PRDX6 protein.
4.Surveillance and early warning index system for schistosomiasis in the middle and lower reaches of the Yangtze River basin
Sanhong JIANG ; Yibiao ZHOU ; Shizhu LI ; Dandan LIN ; Qingwu JIANG ; Liyong WEN ; Shengming LI ; Fei HU ; Benjiao HU ; Jie ZHOU ; Chunli CAO ; Jing XU ; Jianwen XIE ; Changming WU ; Xiaolan YAN ; Weimin XU ; Jun GE ; Guanghui REN ; Xiaoli LIU
Chinese Journal of Endemiology 2025;44(4):259-264
Under the current situation of "low prevalence and low infection" of schistosomiasis in China, and to provide a basis for achieving the goal of eliminating schistosomiasis by 2030 proposed by the Healthy China Action (2019 - 2030) as scheduled, the Hunan Provincial Corps Hospital of the Chinese People's Armed Police Force established a schistosomiasis monitoring and early warning index system based on the previous studies on schistosomiasis early warning index system and the recent literature analysis, combined with the current potential risk factors affecting the transmission and prevalence of schistosomiasis, and organized two rounds of expert consultation and carried out project promotion meetings. The experts reached a consensus on the comprehensiveness and practicability of the index system, aiming to lay a solid foundation for construction of China's schistosomiasis prevention and control early warning system.
5.Mechanism of action of long non-coding RNA AC087388.1 targeting PABPC1 on esophageal squamous cell carcinoma
Han ZHONG ; Yan-lei GE ; Jun-qing GAN ; Ye JIN ; Xuan ZHENG ; Zi-qing LIU ; Guo-gui SUN
Chinese Pharmacological Bulletin 2025;41(5):926-935
Aim To explore the roles and mechanisms of long non-coding RNA(lncRNA)AC087388.1 and poly(A)binding protein cytoplasmic 1(PABPC1)in esophageal squamous cell carcinoma(ESCC).Meth-ods The expression level of AC087388.1 in ESCC tissues and cells was detected by real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)and fluorescence in situ hybridization experiments and its clinical relevance was analyzed.Cell counting kit-8(CCK-8),clone formation,scratch and Transwell inva-sion assays were used to detect the effects of knock-down of AC087388.1 on the cell viability,prolifera-tion,migratory,and invasion of ESCC cells respectively ability,and sub-localization in cells.RNA pull down and Western blot experiments were employed to verify the interaction between AC087388.1 and PABPC1 in ESCC cells.Salvage experiments were performed to detect the effect of AC087388.1 targeting PABPC1 on ESCC cells.Results AC087388.1 was highly ex-pressed in ESCC tissues and cells and positively corre-lated with clinical stage of ESCC patients,mainly local-ized in cytoplasm.Knockdown AC087388.1 inhibited ESCC cell viability,proliferation,migration and inva-sionability.PABPC1 was selected from the results of RNA Pull Down-MS experiments for subsequent experi-ments,and AC087388.1 was verified to bind to PAB-PC1 by RNA Pull Down and Western blot experiments.Overexpression of AC087388.1 was verified by rescue experiment to reverse the effects of knockdown of PAB-PC1 on ESCC cell viability,proliferation,migration and invasion.Conclusions High expression of AC087388.1 correlates with clinical stage and may be a risk factor for ESCC progression.AC087388.1 pro-motes the cell viability,proliferation,migration and in-vasive ability of ESCC cells by targeting PABPC1,which may be a novel biomarker for the diagnosis and treatment of ESCC.
6.circ_0071653 targeted miR-197-3p regulates proliferation and metastasis of esophageal squamous cell carcinoma
Xuan ZHENG ; Yan-lei GE ; Jun-qing GAN ; Ye JIN ; Yi-shuang CUI ; Ya-nan WU ; Zi-qing LIU ; Guo-gui SUN
Chinese Pharmacological Bulletin 2025;41(5):888-898
Aim To investigate the effects of circ_0071653 targeting miR-197-3p on the proliferation and metastasis of esophageal squamous cell carcinoma(ES-CC)cells.Methods The circular structure of circ_0071653 was confirmed by Sanger sequencing and ribo-nuclease R tolerance experiments.Real-time quantita-tive polymerase chain reaction(RT-qPCR)and tissue fluorescence in situ hybridization assay were performed to detect the circ_0071653 expression levels and ana-lyze its clinical relevance.Cell fluorescence in situ hy-bridization and nuclear cytoplasmic separation assays were used to verify the subcellular localization of circ_0071653 and miR-197-3p.Bioinformatics analysis,dual luciferase reporter gene and RT-qPCR assays were conducted to validate the interactions between circ_0071653 and miR-197-3p.Moreover,the cell counting Kit-8(CCK-8),colony formation,scratch,Transwell invasion and subcutaneous tumor formation in nude mice assays were used to evaluate the effects of circ_0071653 and miR-197-3p on cell viability,prolifera-tion,migration,and invasion and in vivo tumorigenesi-sability.Results Circ_0071653 was a circular RNA,which showed high expression in ESCC cell lines and tissues.The expression of circ_0071653 was signifi-cantly correlated with lymph node metastasis and clini-cal stage of ESCC patients.Circ_0071653 and miR-197-3p were mainly localized in the cytoplasm.The databases predict that circ_0071653 had complementa-ry binding sites with miR-197-3p,and their binding were confirmed by dual luciferase reporter geneand RT-qPCR assays.Moreover,the activity,proliferation,migration,invasion and in vivo tumorigenesis abilities of ESCC cells were significantly reduced after knocking down circ_0071653,and this effect could be reversed by downregulating the expression of miR-197-3p.Con-clusions Circ_0071653 promotes the malignant pro-gression of ESCC through targeted regulation of miR-197-3p.
7.Expert consensus on the diagnosis and treatment of cemental tear.
Ye LIANG ; Hongrui LIU ; Chengjia XIE ; Yang YU ; Jinlong SHAO ; Chunxu LV ; Wenyan KANG ; Fuhua YAN ; Yaping PAN ; Faming CHEN ; Yan XU ; Zuomin WANG ; Yao SUN ; Ang LI ; Lili CHEN ; Qingxian LUAN ; Chuanjiang ZHAO ; Zhengguo CAO ; Yi LIU ; Jiang SUN ; Zhongchen SONG ; Lei ZHAO ; Li LIN ; Peihui DING ; Weilian SUN ; Jun WANG ; Jiang LIN ; Guangxun ZHU ; Qi ZHANG ; Lijun LUO ; Jiayin DENG ; Yihuai PAN ; Jin ZHAO ; Aimei SONG ; Hongmei GUO ; Jin ZHANG ; Pingping CUI ; Song GE ; Rui ZHANG ; Xiuyun REN ; Shengbin HUANG ; Xi WEI ; Lihong QIU ; Jing DENG ; Keqing PAN ; Dandan MA ; Hongyu ZHAO ; Dong CHEN ; Liangjun ZHONG ; Gang DING ; Wu CHEN ; Quanchen XU ; Xiaoyu SUN ; Lingqian DU ; Ling LI ; Yijia WANG ; Xiaoyuan LI ; Qiang CHEN ; Hui WANG ; Zheng ZHANG ; Mengmeng LIU ; Chengfei ZHANG ; Xuedong ZHOU ; Shaohua GE
International Journal of Oral Science 2025;17(1):61-61
Cemental tear is a rare and indetectable condition unless obvious clinical signs present with the involvement of surrounding periodontal and periapical tissues. Due to its clinical manifestations similar to common dental issues, such as vertical root fracture, primary endodontic diseases, and periodontal diseases, as well as the low awareness of cemental tear for clinicians, misdiagnosis often occurs. The critical principle for cemental tear treatment is to remove torn fragments, and overlooking fragments leads to futile therapy, which could deteriorate the conditions of the affected teeth. Therefore, accurate diagnosis and subsequent appropriate interventions are vital for managing cemental tear. Novel diagnostic tools, including cone-beam computed tomography (CBCT), microscopes, and enamel matrix derivatives, have improved early detection and management, enhancing tooth retention. The implementation of standardized diagnostic criteria and treatment protocols, combined with improved clinical awareness among dental professionals, serves to mitigate risks of diagnostic errors and suboptimal therapeutic interventions. This expert consensus reviewed the epidemiology, pathogenesis, potential predisposing factors, clinical manifestations, diagnosis, differential diagnosis, treatment, and prognosis of cemental tear, aiming to provide a clinical guideline and facilitate clinicians to have a better understanding of cemental tear.
Humans
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Dental Cementum/injuries*
;
Consensus
;
Diagnosis, Differential
;
Cone-Beam Computed Tomography
;
Tooth Fractures/therapy*
8.Expert consensus on the treatment of oral diseases in pregnant women and infants.
Jun ZHANG ; Chenchen ZHOU ; Liwei ZHENG ; Jun WANG ; Bin XIA ; Wei ZHAO ; Xi WEI ; Zhengwei HUANG ; Xu CHEN ; Shaohua GE ; Fuhua YAN ; Jian ZHOU ; Kun XUAN ; Li-An WU ; Zhengguo CAO ; Guohua YUAN ; Jin ZHAO ; Zhu CHEN ; Lei ZHANG ; Yong YOU ; Jing ZOU ; Weihua GUO
International Journal of Oral Science 2025;17(1):62-62
With the growing emphasis on maternal and child oral health, the significance of managing oral health across preconception, pregnancy, and infancy stages has become increasingly apparent. Oral health challenges extend beyond affecting maternal well-being, exerting profound influences on fetal and neonatal oral development as well as immune system maturation. This expert consensus paper, developed using a modified Delphi method, reviews current research and provides recommendations on maternal and child oral health management. It underscores the critical role of comprehensive oral assessments prior to conception, diligent oral health management throughout pregnancy, and meticulous oral hygiene practices during infancy. Effective strategies should be seamlessly integrated across the life course, encompassing preconception oral assessments, systematic dental care during pregnancy, and routine infant oral hygiene. Collaborative efforts among pediatric dentists, maternal and child health workers, and obstetricians are crucial to improving outcomes and fostering clinical research, contributing to evidence-based health management strategies.
Humans
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Pregnancy
;
Female
;
Infant
;
Consensus
;
Mouth Diseases/therapy*
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Pregnancy Complications/therapy*
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Oral Health
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Infant, Newborn
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Delphi Technique
;
Oral Hygiene
10.Cross-Sectional Analysis of the Relationship Between Bedtime Procrastination and Fear of Missing Out and the Effect of Exercise Intervention.
Jun-Ge PENG ; Meng-Ying ZHANG ; Jiang XIAO ; Kai-Xin LI ; Yue ZHAO ; Yan LI
Acta Academiae Medicinae Sinicae 2025;47(2):175-181
Objective To explore the relationship between bedtime procrastination and fear of missing out and the intervention effect of aerobic exercise on bedtime procrastination,so as to provide a theoretical basis and practical reference for remedying bedtime procrastination.Methods Totally 988 students were selected through random sampling and then surveyed with the bedtime procrastination scale and the fear of missing out scale.Correlation and regression analyses were performed to explore the relationship between bedtime procrastination and fear of missing out.A total of 36 students were recruited from the 988 students to participate in the exercise intervention and they were assigned into an exercise group and a control group by the random number table method,with 18 students in each group.The exercise group performed aerobic exercise for 12 weeks,while the control group maintained daily activities.The participants' scores on the bedtime procrastination scale and the fear of missing out scale were recorded before and after the intervention and compared.Results The fear of missing out was positively correlated with bedtime procrastination among college students(r=0.214,P<0.001),and it was an important predictive factor for bedtime procrastination(β=0.241,P<0.001).After the intervention,the scores of bedtime procrastination scale decreased in the exercise group(t=2.277,P=0.036),while there was no significant difference in the scores of the control group before and after intervention(t=-0.787,P=0.442).Conclusions A high level of fear of missing out indicates severe bedtime procrastination.And 12-week exercise intervention could remedy bedtime procrastination.
Humans
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Fear
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Exercise
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Male
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Female
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Procrastination
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Young Adult
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Cross-Sectional Studies
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Students/psychology*
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Adult
;
Surveys and Questionnaires
;
Adolescent

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