This study aimed to explore the mechanisms and molecular targets of total flavones of Abelmoschus manihot(TFA) plus empagliflozin(EM) in attenuating diabetic tubulopathy(DT) by targeting mitochondrial homeostasis and pyroptosis-apoptosis-necroptosis(PANoptosis). In the in vivo study, the authors established the DT rat models through a combination of uninephrectomy, administration of streptozotocin via intraperitoneal injections, and exposure to a high-fat diet. Following modeling successfully, the DT rat models received either TFA, EM, TFA+EM, or saline(as a vehicle) by gavage for eight weeks, respectively. In the in vitro study, the authors subjected the NRK52E cells with or without knock-down Z-DNA binding protein 1(ZBP1) to a high-glucose(HG) environment and various treatments including TFA, EM, and TFA+EM. In the in vivo and in vitro studies, The authors investigated the relative characteristics of renal tubular injury and renal tubular epithelial cells damage induced by reactive oxygen species(ROS), analyzed the relative characteristics of renal tubular PANoptosis and ZBP1-mediatted PANoptosis in renal tubular epithelial cells, and compared the relative characteristics of the protein expression levels of marked molecules of mitochondrial fission in the kidneys and mitochondrial homeostasis in renal tubular epithelial cells, respectively. Furthermore, in the network pharmacology study, the authors predicted and screened targets of TFA and EM using HERB and SwissTargetPrediction databases; The screened chemical constituents and targets of TFA and EM were constructed the relative network using Cytoscape 3.7.2 network graphics software; The relative targets of DT were integrated using OMIM and GeneCards databases; The intersecting targets of TFA, EM, and DT were enriched and analyzed signaling pathways by Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG) software using DAVID database. In vivo study results showed that TFA+EM could improve renal tubular injury, the protein expression levels and characteristics of key signaling molecules in PANoptosis pathway in the kidneys, and the protein expression levels of marked molecules of mitochondrial fission in the kidneys. And that, the ameliorative effects in vivo of TFA+EM were both superior to TFA or EM. Network pharmacology study results showed that TFA+EM treated DT by regulating the PANoptosis signaling pathway. In vitro study results showed that TFA+EM could improve ROS-induced cell injury, ZBP1-mediatted PANoptosis, and mitochondrial homeostasis in renal tubular epithelial cells under a state of HG, including the protein expression levels of marked molecules of mitochondrial fission, mitochondrial ultrastructure, and membrane potential level. And that, the ameliorative effects in vitro of TFA+EM were both superior to TFA or EM. More importantly, using the NRK52E cells with knock-down ZBP1, the authors found that, indeed, ZBP1 was mediated PANoptosis in renal tubular epithelial cells as an upstream factor. In addition, TFA+EM could regulate the protein expression levels of marked signaling molecules of PANoptosis by targeting ZBP1. In summary, this study clarified that TFA+EM, different from TFA or EM, could attenuate DT with multiple targets by ameliorating mitochondrial homeostasis and inhibiting ZBP1-mediated PANoptosis. These findings provide the clear pharmacological evidence for the clinical treatment of DT with a novel strategy of TFA+EM, which is named "coordinated traditional Chinese and western medicine".
Animals ; Rats ; Mitochondria/metabolism* ; Benzhydryl Compounds/administration & dosage* ; Glucosides/administration & dosage* ; Abelmoschus/chemistry* ; Male ; Homeostasis/drug effects* ; Flavones/administration & dosage* ; Rats, Sprague-Dawley ; Diabetic Nephropathies/physiopathology* ; Drugs, Chinese Herbal/administration & dosage* ; DNA-Binding Proteins/genetics* ; Humans ; Apoptosis/drug effects*

Objective To explore the effect of project-based learning on the training of sharp injury prevention skills in the pre-job training for nursing interns and its impact on the incidence of sharp injuries during internship.Methods Two classes(class A and B),majoring in nursing at Medical College of Nanchang Institute of Technology in March 2023 were selected as the research subjects by convenience sampling method.Coin flipping method was used to designate class A as the routine group and class B as the intervention group.The routine group received rou-tine pre-job training which mainly focused on retrospective intensive training on nursing operation skills.On this ba-sis,the intervention group integrated project-based learning as compensation education on sharp injury prevention skills.Kirkpatrick's four level training evaluation model was adopted to comprehensively evaluate the educational effectiveness at the four progressive levels of"reaction,learning,behavior,and outcome"at corresponding stages.Results 56 nursing interns were included in the class A routine group and class B intervention group,respectively.The course evaluation score(128.67±4.39 vs 117.28±6.55),needlestick protection knowledge cognition score(109.11±4.38 vs 96.44±6.72),safe injection behavior score(38.45±4.91 vs 32.30±5.62),occupational iden-tity score(58.02±8.55 vs 51.77±15.86),and job competency score(82.59±13.35 vs 75.61±15.09)of nursing interns in the intervention group were all higher than those in the routine group,differences were all statistically sig-nificant(all P＜0.05).The incidence of sharp injuries(19.64％ vs 57.14％)and the average frequency of occu-rrence(1.45 vs 2.13)in nursing interns in the intervention group were both lower than those in the control group.The case intervention rate(87.50％ vs 45.59％)and case reporting rate(93.75％vs 32.35％)after sharp injury were both higher than those in the routine group,and the differences were both statistically significant(both P＜0.05).Conclusion Introducing project-based learning in pre-job training for nursing interns can effectively improve their mastery of protection skills,reduce the incidence of sharp injuries during internships,and have important prac-tical value in cultivating their occupational protection abilities.

Objective:To investigate the effect of oleanolic acid(OA)on inflammatory injury of pancreatic acinar cells in-duced by lipopolysaccharide(LPS)by regulating stromal cell-derived factor-1(SDF-1)/CXC chemokine receptor 4(CXCR4)signal axis.Methods:Rat pancreatic acinar cells AR42J were treated with 5,10,20 and 40 μmol/L OA,and the cell viability was detected using CCK-8 method to determine the optimal dosage for subsequent experimental OA treatment;AR42J cells were randomly divided into control group(NC),LPS group(10 mg/L LPS),OA low-dose group(OA-L group,10 mg/L LPS+5 μmol/L OA),OA medium dose group(OA-M group,10 mg/L LPS+10 μmol/L OA),OA high-dose group(OA-H group,10 mg/L LPS+20 μmol/L OA),Rr-SDF-1 activator(SDF-1)group(10 mg/L LPS+20 ng/ml Rr-SDF-1)and OA-H+Rr-SDF-1 group(10 mg/L LPS+20 μmol/L OA+20 ng/ml Rr-SDF-1).CCK-8 method and EdU staining were applied to detect proliferation of AR42J cells;flow cytometry was applied to detect apoptosis of AR42J cells;activity of superoxide dismutase(SOD)and content of malondialdehyde(MDA)in cell supernatant were detected by spectrophotometry;ELISA was applied to detect levels of IL-6 and TNF-α in cell supernatant;Western blot was applied to detect expressions of CyclinD1,Bcl-2 associated X protein(Bax),SDF-1 and CXCR4 proteins in cells.Results:5,10 and 20 μmol/L OA were selected as the low,medium and high doses for subsequent treatment of AR42J cells;compared with NC group,A450 value,EdU positive cell rate,SOD activity,and CyclinD1 protein expression in LPS group decreased,while apoptosis rate,MDA content,IL-6,TNF-α levels,Bax,SDF-1 and CXCR4 protein expressions increased(P<0.05);compared with LPS group,A450 value,EdU positive cell rate,SOD activity,and CyclinD1 protein expression in OA-L group,OA-M group and OA-H group increased,while apoptosis rate,MDA content,IL-6,TNF-α levels,Bax,SDF-1 and CXCR4 protein expressions reduced,the change trends of corre-sponding indicators in Rr-SDF-1 group was opposite to the above(P<0.05);compared with OA-H group,A450 value,EdU positive cell rate,SOD activity,and CyclinD1 protein expression in OA-H+Rr-SDF-1 group decreased,while apoptosis rate,MDA content,IL-6,TNF-α levels,Bax,SDF-1 and CXCR4 protein expressions increased(P<0.05).Conclusion:OA may alleviate LPS induced inflam-matory damage to pancreatic acinar cells in rats by inhibiting the SDF-1/CXCR4 pathway.

Objective To explore the effect of project-based learning on the training of sharp injury prevention skills in the pre-job training for nursing interns and its impact on the incidence of sharp injuries during internship.Methods Two classes(class A and B),majoring in nursing at Medical College of Nanchang Institute of Technology in March 2023 were selected as the research subjects by convenience sampling method.Coin flipping method was used to designate class A as the routine group and class B as the intervention group.The routine group received rou-tine pre-job training which mainly focused on retrospective intensive training on nursing operation skills.On this ba-sis,the intervention group integrated project-based learning as compensation education on sharp injury prevention skills.Kirkpatrick's four level training evaluation model was adopted to comprehensively evaluate the educational effectiveness at the four progressive levels of"reaction,learning,behavior,and outcome"at corresponding stages.Results 56 nursing interns were included in the class A routine group and class B intervention group,respectively.The course evaluation score(128.67±4.39 vs 117.28±6.55),needlestick protection knowledge cognition score(109.11±4.38 vs 96.44±6.72),safe injection behavior score(38.45±4.91 vs 32.30±5.62),occupational iden-tity score(58.02±8.55 vs 51.77±15.86),and job competency score(82.59±13.35 vs 75.61±15.09)of nursing interns in the intervention group were all higher than those in the routine group,differences were all statistically sig-nificant(all P＜0.05).The incidence of sharp injuries(19.64％ vs 57.14％)and the average frequency of occu-rrence(1.45 vs 2.13)in nursing interns in the intervention group were both lower than those in the control group.The case intervention rate(87.50％ vs 45.59％)and case reporting rate(93.75％vs 32.35％)after sharp injury were both higher than those in the routine group,and the differences were both statistically significant(both P＜0.05).Conclusion Introducing project-based learning in pre-job training for nursing interns can effectively improve their mastery of protection skills,reduce the incidence of sharp injuries during internships,and have important prac-tical value in cultivating their occupational protection abilities.

Objective:To investigate the effect of oleanolic acid(OA)on inflammatory injury of pancreatic acinar cells in-duced by lipopolysaccharide(LPS)by regulating stromal cell-derived factor-1(SDF-1)/CXC chemokine receptor 4(CXCR4)signal axis.Methods:Rat pancreatic acinar cells AR42J were treated with 5,10,20 and 40 μmol/L OA,and the cell viability was detected using CCK-8 method to determine the optimal dosage for subsequent experimental OA treatment;AR42J cells were randomly divided into control group(NC),LPS group(10 mg/L LPS),OA low-dose group(OA-L group,10 mg/L LPS+5 μmol/L OA),OA medium dose group(OA-M group,10 mg/L LPS+10 μmol/L OA),OA high-dose group(OA-H group,10 mg/L LPS+20 μmol/L OA),Rr-SDF-1 activator(SDF-1)group(10 mg/L LPS+20 ng/ml Rr-SDF-1)and OA-H+Rr-SDF-1 group(10 mg/L LPS+20 μmol/L OA+20 ng/ml Rr-SDF-1).CCK-8 method and EdU staining were applied to detect proliferation of AR42J cells;flow cytometry was applied to detect apoptosis of AR42J cells;activity of superoxide dismutase(SOD)and content of malondialdehyde(MDA)in cell supernatant were detected by spectrophotometry;ELISA was applied to detect levels of IL-6 and TNF-α in cell supernatant;Western blot was applied to detect expressions of CyclinD1,Bcl-2 associated X protein(Bax),SDF-1 and CXCR4 proteins in cells.Results:5,10 and 20 μmol/L OA were selected as the low,medium and high doses for subsequent treatment of AR42J cells;compared with NC group,A450 value,EdU positive cell rate,SOD activity,and CyclinD1 protein expression in LPS group decreased,while apoptosis rate,MDA content,IL-6,TNF-α levels,Bax,SDF-1 and CXCR4 protein expressions increased(P<0.05);compared with LPS group,A450 value,EdU positive cell rate,SOD activity,and CyclinD1 protein expression in OA-L group,OA-M group and OA-H group increased,while apoptosis rate,MDA content,IL-6,TNF-α levels,Bax,SDF-1 and CXCR4 protein expressions reduced,the change trends of corre-sponding indicators in Rr-SDF-1 group was opposite to the above(P<0.05);compared with OA-H group,A450 value,EdU positive cell rate,SOD activity,and CyclinD1 protein expression in OA-H+Rr-SDF-1 group decreased,while apoptosis rate,MDA content,IL-6,TNF-α levels,Bax,SDF-1 and CXCR4 protein expressions increased(P<0.05).Conclusion:OA may alleviate LPS induced inflam-matory damage to pancreatic acinar cells in rats by inhibiting the SDF-1/CXCR4 pathway.

AIM To study the neoflavonoids from Dalbergia cochinchinensis Pierre ex Laness and their anti-hypoxia/reoxygenation injury activities on H9c2 myocardial cells.METHODS The 70%ethanol extract from D.cochinchinensis was isolated and purified by silica gel,Sephadex LH-20 and reverse-preparative HPLC,then the structures of obtained compounds were identified by physicochemical properties and spectral data.The CCK-8 method was used to detect their activities on H9c2 cells and protective effects on hypoxia-reoxygenation injury of H9c2 cells,and their structure-activity relationship was analyzed.RESULTS Twelve compounds were isolated and identified as latifolin(1),5-O-methyllatifolin(2),mimosifoliol(3),5-O-methydalbergiphenol(4),dalbergiphenol(5),cearoin(6),2,4-dihydroxy-5-methoxy-benzophenone(7),2-hydroxy-4,5-dimethoxybenzophenone(8),melannoin(9),2,2′,5-trihydroxy-4-methoxybenzophenone(10),dalbergin(11),4-methoxydalbergione(12).The dalbergiphenols and dalbergins had little toxicity to H9c2 cells,and dalbergiphenols had strong activity against hypoxia-reoxygenation injury of H9c2 cells.CONCLUSION Compound 8 is a new natural product.Compounds 4,9 are isolated from this plant for the first time.Dalbergiphenols may be the main neoflavonoids against hypoxia-reoxygenation injury of H9c2 cells.

A ratiometric fluorescent probe([Eu(Hpfdap)3(CTMR-Phen-N)])was designed and synthesized for detection of singlet oxygen(1O2)with dimethyl anthracyl-containing β-diketone compound 1,1,1,2-pentafluoro-5-(9',10'-dimethyl-2'-anthracene)-3,5-pentanedione(Hpfdap)as recognition ligand and 5-amino-1,10-phenanthroline conjugated with Rhodamine as auxiliary ligand(CTMR-Phen-N).The results of spectral analysis showed that the europium(Ⅲ)complex had almost no fluorescence due to the presence of anthracene ring in the probe.After reaction with 1O2,the anthracene ring was oxidized to endoperoxide,and the fluorescence of the europium(Ⅲ)complex at 614 nm was restored,followed by the increase of quantum yield from 1.60%to 25.16%.The fluorescence intensity of Rhodamine at 575 nm remained unchanged before and after the reaction with 1O2.Therefore,the ratio of fluorescence intensity at 614 nm and 575 nm could be used to measure 1O2 by the developed rationmetric fluorescence method.The probe showed good selectivity for 1O2,and the detection limit(S/N=3)was as low as 3 μmol/L.The fluorescence imaging results demonstrated that the probe had low cytotoxicity and good membrane permeability,and could be applied to fluorescence imaging of 1O2 in Hela cells.

Objective:To investigate impacts of usnic acid(UA)on malignant behavior of gastric cancer cells by regulating the chemokine(C-C motif)ligand 2(CCL2)-CCL2 receptor(CCR2)signal axis.Methods:SGC-7901 cells,a well growing human gastric cancer cell line,were treated with different concentrations of UA,which were grouped into low concentration(UA-L)group(62.5 μmol/L UA),medium concentration(UA-M)group(125 μmol/L UA)and high concentration(UA-H)group(250 μmol/L UA);meantime,the cells were transfected with CCL2 overexpression vector(pc DNA3.1 CCL2),empty vector(pc DNA3.1),silenced CCL2(si CCL2)and negative control(si control),and SGC-7901 cells were treated with 250 μmol/L UA,labeled as UA-H+pc DNA3.1 CCL2 group,UA-H+pc DNA3.1 group,UA-H+si control group and UA-H+si CCL2 group,another untreated SGC-7901 cells were taken as the control group.Flow cytometry,MTT and qRT-PCR were applied to detect cell apoptosis,proliferation,and expres-sion levels of CCL2 and CCR2 mRNA;Western blot was applied to detect expression levels of PD-L1,apoptotic protein(Bax),proli-ferative protein(CyclinD1,CCL2,CCR2)and immune escape related protein(B7H1);after co-culturing with CD8+T cells isolated and cultured in vitro,ELISA was applied to detect levels of IL-4,IFN-γ and IL-10 in the supernatant.Gastric cancer cells in each group were co-cultured with activated peripheral blood mononuclear cell(PBMC)1∶1 for 72 hours,and the sensitivity of gastric can-cer cells in each group to T-cell-mediated killing was compared.Results:Compared with control group,cell proliferation rate,IL-10 level,CyclinD1,PD-L1,CCL2,CCR2 and B7H1 protein and mRNA expressions,cell counts after co-culturing with activated PBMC 1∶1 for 72 hours in UA-L group,UA-M group and UA-H group were obviously reduced,while apoptosis rate,IL-4 and IFN-γ levels,Bax protein expression were obviously increased(P<0.05);compared with UA-H+pc DNA3.1 group,cell proliferation rate,IL-10 level,CyclinD1,PD-L1,CCL2,CCR2 protein and mRNA expressions,cell counts after co-culturing with activated PBMC 1∶1 for 72 hours in UA-H+pc DNA3.1 CCL2 group were obviously increased,while apoptosis rate,IL-4 and IFN-γ levels,and Bax protein ex-pression were obviously reduced(P<0.05);compared with UA-H+si control group,cell proliferation rate,IL-10 level,CyclinD1,PD-L1,CCL2,CCR2 and B7H1 protein and mRNA expressions,cell counts after co-culturing with activated PBMC 1∶1 for 72 hours in UA-H+si CCL2 group were obviously reduced,while apoptosis rate,IL-4 and IFN-γ levels and Bax protein expression were obviously increased(P<0.05).Conclusion:UA can inhibit gastric cancer cells proliferation,immune escape,and induce apoptosis,which may be related to the inhibition of the CCL2-CCR2 signaling axis.

At present, China's traditional Chinese medicine(TCM) industry is developing rapidly with the support of modern science and technology. While promoting economic development and improving national health, it has brought multiple environmental problems. Under the "dual carbon" goals, the ecological fine manufacturing of TCM may become one of the breakthroughs for the TCM industry to practice low-carbon economy. From the perspective of low-carbon economy and considering the current situation of TCM pharmaceutical manufacturing, this paper analyzes the problems and shortcomings of the TCM pharmaceutical industry. In view of the key factors influencing the quality of TCM preparations under the ecological fine manufacturing mode, this paper proposes a practical and feasible selection plan for the ecological fine manufacturing mode of TCM, aiming to provide research ideas and a theoretical basis for the TCM industry in helping to achieve the goals of carbon peaking and carbon neutrality.
Drugs, Chinese Herbal/economics* ; Medicine, Chinese Traditional ; Carbon/analysis* ; China ; Drug Industry/economics*

Objective:To investigate the clinical characteristics and interventions associated with drug-induced anaphylaxis in the emergency infusion room.Methods:Bases on the adverse drug reaction database from the emergency medicine center of the First Affiliated Hospital of Nanjing Medical University, clinical data of patients who experienced drug-induced anaphylaxis in the emergency infusion room between November 2019 and November 2023 were collected, including gender, age, history of previous adverse drug reactions, allergy history, Charlson comorbidity index, medication details, information related to drug-induced anaphylaxis (onset time, clinical manifestations), interventions, outcomes, and follow-up. The clinical characteristics and interventions in these patients were analyzed.Results:During the study period, a total of 398 772 patients in the emergency infusion room in our hospital received intravenous infusion of drugs. Of them, 625 cases developed adverse drug reactions (ADRs) and 75 cases developed drug-induced anaphylaxis, accounting for 0.02% (75/398 772) of the total infusion patients and 12.0% (75/625) of all ADR cases. Of the 75 patients with anaphylaxis, 30 cases (40%) were classified as grade Ⅱ, and 45 cases (60%) as grade Ⅲ, with no grade Ⅳ cases. The most common drugs involved in 75 cases of anaphylaxis were anti-infective drugs (41 cases, 54.7%). Drug-induced anaphylaxis exhibited diverse clinical manifestations, with cardiovascular symptoms being the most common, primarily varying degrees of transient hypotension (67 cases, 89.3%), followed by systemic and neurological symptoms, including profuse sweating (31 cases, 41.3%) and dizziness (28 cases, 37.3%). All 75 patients with anaphylaxis were treated with measures such as discontinuation of medication, replacement of infusion sets, rapid assessment of circulation and respiration, and monitoring of vital signs, of which 65 (86.7%) received rapid intravenous infusion for volume expansion, 6 (8.0%) received intravenous injection of glucocorticoids, 3 (4.0%) received intramuscular injection of 0.5 mg epinephrine, and 2 (2.7%) received antihistamines. All 75 patients showed improvement in symptoms, and no sequelae or deaths were found.Conclusions:In the emergency infusion room, the severity of anaphylaxis is mainly grade Ⅱ and Ⅲ with a good prognosis after timely intervention. The treatment measures mainly focus on rapid intravenous infusion for volume expansion, and the use of epinephrine is relatively low.