OBJECTIVE To screen and quantitatively analyze the quality marker （Q-Marker） associated with anti-renal fibrosis in Shenqi shenshuai mixture （SQSS）， so as to provide references for the analysis of pharmacodynamic substances and new drug transformation of SQSS. METHODS The chemical components of SQSS were characterized by untargeted metabolomics. Combined with network pharmacology， gene expression omnibus （GEO） and connectivity map （CMAP） databases， the anti-renal fibrosis Q-Markers of SQSS were screened. HPLC-MS/MS was applied to determine the contents of Q-Markers in 10 batches of SQSS. RESULTS A total of 1 319 compounds were identified from SQSS via untargeted metabolomics，and 84 active ingredients with anti-renal fibrosis activity were further screened out， such as formononetin. Nine anti-renal fibrosis Q-Markers were obtained by network pharmacology and GEO database， including berberine， quercetin， honokiol， nicotinamide， daidzein， coumarin， kaempferol， formononetin and amygdalin. The quantitative results showed that the average contents of the above components （excluding amygdalin） in 10 batches of SQSS were 2.523， 1.942， 26.848， 1.415， 0.692， 0.171， 0.374， 7.401 μg/mL， respectively. CONCLUSIONS Nine anti-renal fibrosis Q-Markers of SQSS were screened in this study， and the contents of eight among them were determined. The results can provide a basis for elucidating the pharmacodynamic material basis and promoting the new drug transformation of SQSS.

OBJECTIVE To screen and quantitatively analyze the quality marker （Q-Marker） associated with anti-renal fibrosis in Shenqi shenshuai mixture （SQSS）， so as to provide references for the analysis of pharmacodynamic substances and new drug transformation of SQSS. METHODS The chemical components of SQSS were characterized by untargeted metabolomics. Combined with network pharmacology， gene expression omnibus （GEO） and connectivity map （CMAP） databases， the anti-renal fibrosis Q-Markers of SQSS were screened. HPLC-MS/MS was applied to determine the contents of Q-Markers in 10 batches of SQSS. RESULTS A total of 1 319 compounds were identified from SQSS via untargeted metabolomics，and 84 active ingredients with anti-renal fibrosis activity were further screened out， such as formononetin. Nine anti-renal fibrosis Q-Markers were obtained by network pharmacology and GEO database， including berberine， quercetin， honokiol， nicotinamide， daidzein， coumarin， kaempferol， formononetin and amygdalin. The quantitative results showed that the average contents of the above components （excluding amygdalin） in 10 batches of SQSS were 2.523， 1.942， 26.848， 1.415， 0.692， 0.171， 0.374， 7.401 μg/mL， respectively. CONCLUSIONS Nine anti-renal fibrosis Q-Markers of SQSS were screened in this study， and the contents of eight among them were determined. The results can provide a basis for elucidating the pharmacodynamic material basis and promoting the new drug transformation of SQSS.

OBJECTIVE To investigate α1-adrenergic receptors(α1-AR)distribution in mouse tissues and its function on dexmetomidine(DMED)induced toxic effects.METHODS ① Real-time fluorescence quantita-tive PCR was used to detect the relative expression of α1A-AR,α1B-AR,α1D-AR,α2A-AR,α2B-AR and α2C-AR mRNAs in the heart,apical potion of heart,lungs,apical potion of lung,liver,kidneys,abdominal aorta,prefrontal cortex,hippocampus,striatum,brainstem,thalamus,olfactory bulb,and the rest of the brain tissues of the mouse.The relative expression of mRNA were analyzed.② C57BL/6J mice were pretreated with α2 adrenergic receptor antagonist atipamezole ATI(0.005,0.010,0.020,0.025,0.040,0.050 mg·kg-1,im),orα1-adrenoceptor antagonist prazosin(1 mg·kg-1,im)for 15 min,and then DMED(0.20 mg·kg-1,iv)was given to observe the rate of the loss of righting reflex and the immobilization time in mice.③ C57BL/6J mice were treated with DMED(16.38,20.48,25.60,32.00,40.00,and 50.00 mg·kg-1,iv)to observe the lethality of the mice in 24 h.The dose-effect relationship curves of the lethality rate and the half lethal-dose(LD50)were detected.ATI(1,2,4,and 8 mg·kg-1,im)or prazosin(1 mg·kg-1,im)were pretreated 15 min followed by the administration of DMED(25.60 mg·kg-1,iv).The lethality of the mice were recorded for 24 h.HE staining to observe the lung tissue damage in the mice.RESULTS ① The mRNA expression levels of three α1-AR subtype were higher than those of α2-AR subtype.α2A-AR and α2C-AR were highly expressed in the central nervous system.α2B-AR was highly expressed in the brainstem and peripheral tissues.The mRNA expres-sion levels of α1-AR subtypes were higher than those of α2-AR subtypes in heart,apical potion of heart or lung(P＜0.05).② ATI(0.005 to 0.05 mg·kg-1,im)dose dependently antagonized the loss of righting reflex and decreased the immobilization time induced by DMED(0.20 mg·kg-1,iv).In contrast,prazosin(1 mg·kg-1,im)had no effect on the loss of righting reflex induced by DMED(0.20 mg·kg-1,iv).③ The LD50 of DMED in mice was 26.734 mg·kg-1(iv)with a 95％Cl of 23.606-30.000 mg·kg-1.DMED(25.6 mg·kg-1)was selected for subsequent toxicity.ATI(1,2,4,and 8 mg·kg-1,im)did not antagonize the lethality induced by DMED(25.6 mg·kg-1,iv).The high dose of ATI resulted in elevated death rate and accelerated mortality induced by DMED(25.6 mg·kg-1,iv)in the mice.However,prazosin(1 mg·kg-1,im)reduced the lethality of DMED(25.6 mg·kg-1,iv)(P＜0.01).After administration of DMED(25.6 mg·kg-1),the mice lungs showed significant congestion.HE staining of lung tissues revealed obvious vascular hemorrhage,alveolar rupture,and erythrocyte spillage.Prazosin(1 mg·kg-1,im)effectively attenuated the tissue damage in the lungs,but ATI(1 mg·kg-1,im)aggravated the pulmonary hemorrhage.CONCLU-SIONS In cardiopulmonary tissues,the high expression levels of α1 adrenoceptor overactivation,might related with the lethality induced by DMED.

OBJECTIVE To investigate α1-adrenergic receptors(α1-AR)distribution in mouse tissues and its function on dexmetomidine(DMED)induced toxic effects.METHODS ① Real-time fluorescence quantita-tive PCR was used to detect the relative expression of α1A-AR,α1B-AR,α1D-AR,α2A-AR,α2B-AR and α2C-AR mRNAs in the heart,apical potion of heart,lungs,apical potion of lung,liver,kidneys,abdominal aorta,prefrontal cortex,hippocampus,striatum,brainstem,thalamus,olfactory bulb,and the rest of the brain tissues of the mouse.The relative expression of mRNA were analyzed.② C57BL/6J mice were pretreated with α2 adrenergic receptor antagonist atipamezole ATI(0.005,0.010,0.020,0.025,0.040,0.050 mg·kg-1,im),orα1-adrenoceptor antagonist prazosin(1 mg·kg-1,im)for 15 min,and then DMED(0.20 mg·kg-1,iv)was given to observe the rate of the loss of righting reflex and the immobilization time in mice.③ C57BL/6J mice were treated with DMED(16.38,20.48,25.60,32.00,40.00,and 50.00 mg·kg-1,iv)to observe the lethality of the mice in 24 h.The dose-effect relationship curves of the lethality rate and the half lethal-dose(LD50)were detected.ATI(1,2,4,and 8 mg·kg-1,im)or prazosin(1 mg·kg-1,im)were pretreated 15 min followed by the administration of DMED(25.60 mg·kg-1,iv).The lethality of the mice were recorded for 24 h.HE staining to observe the lung tissue damage in the mice.RESULTS ① The mRNA expression levels of three α1-AR subtype were higher than those of α2-AR subtype.α2A-AR and α2C-AR were highly expressed in the central nervous system.α2B-AR was highly expressed in the brainstem and peripheral tissues.The mRNA expres-sion levels of α1-AR subtypes were higher than those of α2-AR subtypes in heart,apical potion of heart or lung(P＜0.05).② ATI(0.005 to 0.05 mg·kg-1,im)dose dependently antagonized the loss of righting reflex and decreased the immobilization time induced by DMED(0.20 mg·kg-1,iv).In contrast,prazosin(1 mg·kg-1,im)had no effect on the loss of righting reflex induced by DMED(0.20 mg·kg-1,iv).③ The LD50 of DMED in mice was 26.734 mg·kg-1(iv)with a 95％Cl of 23.606-30.000 mg·kg-1.DMED(25.6 mg·kg-1)was selected for subsequent toxicity.ATI(1,2,4,and 8 mg·kg-1,im)did not antagonize the lethality induced by DMED(25.6 mg·kg-1,iv).The high dose of ATI resulted in elevated death rate and accelerated mortality induced by DMED(25.6 mg·kg-1,iv)in the mice.However,prazosin(1 mg·kg-1,im)reduced the lethality of DMED(25.6 mg·kg-1,iv)(P＜0.01).After administration of DMED(25.6 mg·kg-1),the mice lungs showed significant congestion.HE staining of lung tissues revealed obvious vascular hemorrhage,alveolar rupture,and erythrocyte spillage.Prazosin(1 mg·kg-1,im)effectively attenuated the tissue damage in the lungs,but ATI(1 mg·kg-1,im)aggravated the pulmonary hemorrhage.CONCLU-SIONS In cardiopulmonary tissues,the high expression levels of α1 adrenoceptor overactivation,might related with the lethality induced by DMED.

Objective:To explore the dynamic change trajectory and influencing factors of postoperative fatigue (POF) in patients with early digestive tract cancer undergoing endoscopic submucosal dissection (ESD), so as to provide theoretical basis for individualized management of such patients.Methods:Using cross-sectional survey and convenient sampling method, the patients who underwent ESD for early cancer of digestive tract in the endoscopic center of the Second Affiliated Hospital of Zhejiang University School of Medicine from January to June 2024 were selected as the research objects. The questionnaires were conducted at 30 min, 24 h, 3 days, 5 days, 7 days after ESD with Christensen's postoperative fatigue score. Spearman test was used to analyze the correlation between pain, insomnia and POF. The latent variable growth model was used to identify the potential categories of POF trajectory, and the influencing factors were analyzed by Logistic regression.Results:A total of 232 patients were finally induded, with ages of 19-94(59.53 ± 13.29) years, including 120 males and 112 femalss. The POF level of patients with early cancer of digestive tract showed a downward trend one week after operation. Three postoperative fatigue trajectories were fitted, 9.05% in moderate or severe fatigue low-speed smooth descent group (C1 group); 32.76% in moderate fatigue first decreased quickly and then slowly group (C2 group) and 58.19% in mild fatigue continuous decline group (C3 group). The result of Logistic regression analysis showed that, compared with C1 group, people without religious beliefs were morelikely to enter C2 group [ β = 1.572, OR = 4.818(1.033 - 22.465), P<0.05]; compared with C3 group, patients with high pain level and severe insomnia degree were likely to enter C1 [ β = 2.621, 0.663, OR were 13.754(2.692 - 70.283) and 1.942(1.429 - 2.638), both P<0.05] and C2 [ β = 2.010, 0.491, OR were 7.464(1.890 - 29.482) and 1.634(1.348 - 1.982), both P<0.05] group. Conclusions:There were three potential types of POF in patients with early digestive tract cancer ESD. Medical staff should pay attention to patients with severe insomnia, intense pain and religious beliefs, and give staged fatigue assessment and individualized intervention, so as to reduce the level of POF and promote rapid recovery.

Hydrogen-rich water (HRW) shows excellent antibacterial, anti-inflammatory, antioxidant, and wound-healing properties and plays a positive role in the treatment of various diseases, such as brain injury, kidney injury, and periodontitis. Current studies found that HRW can inhibit periodontopathogenic biofilm formation, inhibit oral connective tissue and bone tissue destruction, and show anti-inflammatory and antioxidant properties in periodontitis. Additionally, HRW can alleviate periodontal tissue damage by inhibiting oxidative stress and up-regulating the expression of antioxidant enzymes, such as glutathione peroxidase and superoxide dismutase. HRW exerts anti-inflammatory effects by regulating the nuclear factor erythroid 2-related factor 2 and mitogen-activated protein kinase pathways, which are closely associated with inflammation. Additionally, HRW inhibits the expression of inflammatory cytokines, such as interleukins, inhibits the growth and proliferation of bacterial plaque biofilms, and down-regulates glycosyltransferases and glucan-binding proteins to prevent bacterial adhesion and subsequent development of periodontitis. Furthermore, HRW has a positive effect on the expression of various cell growth factors, α-smooth muscle actin, and type I collagen, which promotes wound healing. Current clinical studies have demonstrated the biological safety of HRW (to a certain extent) and reported no adverse reactions. However, most studies on HRW in oral diseases are preclinical in vivo and in vitro studies. Therefore, further clinical studies are required to validate the therapeutic significance and optimal therapeutic regimen of HRW in human periodontitis. This article aims to review the therapeutic role and the underlying mechanisms of HRW in periodontitis.

Objective To summarize the current status of outcome indicators in randomized controlled trials of TCM intervention in pediatric myocarditis,to explore the existing problems,and to provide a reference for the construction of a core set of indicators for the treatment of pediatric myocarditis with TCM.Methods Randomized controlled trial literature on the treatment of pediatric myocarditis with TCM was retrieved from CNKI,Wanfang Data,VIP,SinoMed,PubMed,Embase,Cochrane Library and Web of Science from the establishment of the databases to 16th,Nov.2024.The basic characteristics,diagnostic criteria,TCM evidence,interventions and outcome indicators of the studies were extracted.The risk of bias was assessed for the selected studies using the Cochrane Collaboration's risk of bias tool(RoB 2.0),and the outcome indicators were statistically analyzed using Excel 2019.Results Finally,totally of 250 articles were included in the literature.The outcome indicators were counted to obtain 187 outcome indicators with a cumulative total of 1 540 occurrences,which were categorized into six indicator domains:symptoms and signs,physicochemical testing,TCM symptoms/signs,clinical efficacy evaluation,safety evaluation and quality of life evaluation;among them,the physicochemical testing indicators(112 types,926 times)had the highest frequency of occurrences,followed by the clinical efficacy evaluation(13 types.340 times),and safety evaluation(24 types,193 times).Conclusion The overall quality of randomized controlled trials of TCM treatment of pediatric myocarditis is low,and there are problems with primary and secondary differentiation of outcome indicators and lack of TCM characteristics.There is an urgent need to improve the core set of endpoint indicators that highlight the characteristics of TCM in order to improve the quality of clinical research.

Objective:To explore the dynamic change trajectory and influencing factors of postoperative fatigue (POF) in patients with early digestive tract cancer undergoing endoscopic submucosal dissection (ESD), so as to provide theoretical basis for individualized management of such patients.Methods:Using cross-sectional survey and convenient sampling method, the patients who underwent ESD for early cancer of digestive tract in the endoscopic center of the Second Affiliated Hospital of Zhejiang University School of Medicine from January to June 2024 were selected as the research objects. The questionnaires were conducted at 30 min, 24 h, 3 days, 5 days, 7 days after ESD with Christensen's postoperative fatigue score. Spearman test was used to analyze the correlation between pain, insomnia and POF. The latent variable growth model was used to identify the potential categories of POF trajectory, and the influencing factors were analyzed by Logistic regression.Results:A total of 232 patients were finally induded, with ages of 19-94(59.53 ± 13.29) years, including 120 males and 112 femalss. The POF level of patients with early cancer of digestive tract showed a downward trend one week after operation. Three postoperative fatigue trajectories were fitted, 9.05% in moderate or severe fatigue low-speed smooth descent group (C1 group); 32.76% in moderate fatigue first decreased quickly and then slowly group (C2 group) and 58.19% in mild fatigue continuous decline group (C3 group). The result of Logistic regression analysis showed that, compared with C1 group, people without religious beliefs were morelikely to enter C2 group [ β = 1.572, OR = 4.818(1.033 - 22.465), P<0.05]; compared with C3 group, patients with high pain level and severe insomnia degree were likely to enter C1 [ β = 2.621, 0.663, OR were 13.754(2.692 - 70.283) and 1.942(1.429 - 2.638), both P<0.05] and C2 [ β = 2.010, 0.491, OR were 7.464(1.890 - 29.482) and 1.634(1.348 - 1.982), both P<0.05] group. Conclusions:There were three potential types of POF in patients with early digestive tract cancer ESD. Medical staff should pay attention to patients with severe insomnia, intense pain and religious beliefs, and give staged fatigue assessment and individualized intervention, so as to reduce the level of POF and promote rapid recovery.

Objective To summarize the current status of outcome indicators in randomized controlled trials of TCM intervention in pediatric myocarditis,to explore the existing problems,and to provide a reference for the construction of a core set of indicators for the treatment of pediatric myocarditis with TCM.Methods Randomized controlled trial literature on the treatment of pediatric myocarditis with TCM was retrieved from CNKI,Wanfang Data,VIP,SinoMed,PubMed,Embase,Cochrane Library and Web of Science from the establishment of the databases to 16th,Nov.2024.The basic characteristics,diagnostic criteria,TCM evidence,interventions and outcome indicators of the studies were extracted.The risk of bias was assessed for the selected studies using the Cochrane Collaboration's risk of bias tool(RoB 2.0),and the outcome indicators were statistically analyzed using Excel 2019.Results Finally,totally of 250 articles were included in the literature.The outcome indicators were counted to obtain 187 outcome indicators with a cumulative total of 1 540 occurrences,which were categorized into six indicator domains:symptoms and signs,physicochemical testing,TCM symptoms/signs,clinical efficacy evaluation,safety evaluation and quality of life evaluation;among them,the physicochemical testing indicators(112 types,926 times)had the highest frequency of occurrences,followed by the clinical efficacy evaluation(13 types.340 times),and safety evaluation(24 types,193 times).Conclusion The overall quality of randomized controlled trials of TCM treatment of pediatric myocarditis is low,and there are problems with primary and secondary differentiation of outcome indicators and lack of TCM characteristics.There is an urgent need to improve the core set of endpoint indicators that highlight the characteristics of TCM in order to improve the quality of clinical research.

OBJECTIVE To establish the cells stably co-expressing α1A-adrenergic receptor(α1A-AR)and enhanced green fluorescent protein(EGFP)tagged nuclear factor of activated T cells 2(NFAT2)(EGFP-NFAT2)in U2OS cells.METHODS ① The pcDNA3.1-α1-AR-3×FLAG recombinant plasmid was transfected into U2OS-EGFP-NFAT2 cells.The transfected cells were selected by hygromycin B(Hygro-B,200 mg·L-1),and screened by EGFP-NFAT2 nuclear translocation assay after α1A-AR agonist norepinephrine(NE)treatment of 30 min.② The mRNA and protein expression levels of α1A-AR in the selected U2OS-EGFP-NFAT2-α1A-AR cells were examined by real-time quantitative PCR(RT-qPCR)and Western blotting.③ U2OS-EGFP-NFAT2-α1A-AR cells were treated with NE(10-8-10-5 mol·L-1)or dexmedetomidine(DMED,10-8.8-10-5 mol·L-1),respectively,for 30 min.EGFP-NFAT2 nuclear translo-cation was detected by high throughout screening assay.④ The U2OS-EGFP-NFAT2-α1A-AR cells were divided into the solvent control group,α1-AR antagonist naftopidill(1 μmol·L-1)group,NE(1 μmol·L-1)group and naftopidill+NE(co-incubation with naftopidill 1 μmol·L-1 and NE 1 μmol·L-1)group,α2-AR antagonist atipamezole(ATI,0.1 μmol·L-1)group,α 2-AR agonist DMED(0.1 μmol·L-1)group,and ATI+DMED(co-incubation with ATI 1 μmol·L-1 and DMED 0.1 μmol·L-1)group.The drug incubation time was 30 min.EGFP-NFAT2 nuclear translocation was abserved via a high throughout screening system to validate the α1A-AR function in U2OS-EGFP-NFAT2-α1A-AR cells.RESULTS ① There were 58 cell strains expressing α1A-AR in U2OS-EGFP-NFAT2 cells by EGFP-NFAT2 nuclear translocation assay.Among these cells,cells No 50 had the highest nuclear translocation function.The α1A-AR mRNA expression of cells No 50 in 5-20 generations were detected by RT-qPCR and were about 500-800 times that of U2OS-EGFP-NFAT2 cells.② The protein band of α1A-AR was also detected in cells No 50,but no band of α1A-AR was detected in U2OS-EGFP-NFAT2 cells by Western blotting.③ NE and DMED increased the relative translocation nuclear index in U2OS-EGFP-NFAT2-α1A-AR cells with ED50 5.94×10-7 and 6.15×10-8 mol·L-1,respectively.④ EGFP-NFAT2 nuclear translocation was significant in U2OS-EGFP-NFAT2-α1A-AR cells after NE addition compared with the solvent control or the naftopidill groups(P<0.01).The EGFP-NFAT2 nuclear translocation in the naftopidill+NE group was significantly decreased compared with the NE group(P<0.01).DMED significantly increased the EGFP-NFAT2 nuclear translocation compared with solvent control or the ATI groups(P<0.01).The EGFP-NFAT2 nuclear translocation in the ATI+DMED group was similar to that of the DMED group.The EGFP-NFAT2 nuclear translocation in the naftopidill+DMED group was decreased significantly compared with the DMED group(P<0.01).CONCLUSION U2OS-EGFP-NFAT2-α1A-AR cells stably co-exrepssing α1A-AR and EGFP-NFAT2 are established,which can be used for high throughout screening of biased chemicals and studies on the mechanism of α1A-AR.