Objective To construct and evaluate a mouse model of renal fibrosis(RF)combined with Qi deficiency and dampness stasis,and investigate the changes in protein and metabolic pathways using multiomics.Methods Twenty-four C57BL/6J mice were divided randomly into normal(N),model(M),and RF and syndrome combined groups(BZ)(n=8/group).The experiment lasted 6 weeks.A mouse model of RF with Qi deficiency and dampness stasis was established by "cyclosporine A+high-fat diet+swimming exhaustion+constant temperature and humidity".The model was evaluated by detecting general signs,renal function,tongue RGB(red,green,blue)values,hemorheology indexes,blood lipids,and inflammation and oxidation indexes,combined with hematoxylin and eosin,Masson,periodic acid-Schiff,and Oil red O staining,terminal deoxynucleotidyl transferase dUTP nick end labeling apoptosis,and transforming growth factor-β immunofluorescence analysis of renal tissue.Differential proteins and metabolites were screened by renal proteomics combined with serum metabolomics and subjected to pathway enrichment analysis.Results Body mass of mice in the BZ group began to decline at week 3(P＜0.05)and decreased significantly at week 4(P＜0.01),while food and water consumption decreased,the fur became messy and less glossy,mood and activity decreased,and stools became watery.Serum creatinine,blood urea nitrogen,urine albumin-creatinine ratio,and N-acetyl-beta-glucosaminidase(NAG)were significantly higher in the BZ group compared with those in the N group(P＜0.05,P＜0.01),and serum creatinine and NAG levels were significantly different compared with those in the M group.The R value of tongue images was significantly lower in the BZ group compared with that in the N group(P＜0.01),while the B value was significantly higher(P＜0.05).The viscosity of the whole blood multi-shear rate and the hematocrit were higher in the BZ group compared with those in the N and M groups,and the platelet volume was higher than in the N group(P＜0.05,P＜0.01).Total cholesterol,low-density lipoprotein cholesterol,C-reactive protein,interleukin-6,and malondialdehyde levels were significantly increased in the BZ group compared with those in the N and M groups(P＜0.01),and superoxide dismutase activity was significantly decreased compared with that in the N group(P＜0.05).Renal tubule vacuolation,inflammatory cell infiltration,glomerular basement membrane thickening,collagen fiber hyperplasia,and lipid accumulation were evident,and renal cell apoptosis and transforming growth factor-β deposition were increased in the BZ group.There were 299 differential proteins in the BZ and N groups,including 180 up-regulated and 119 down-regulated proteins,and 323 differential metabolites,including 205 up-regulated and 118 down-regulated.Primary bile acid biosynthesis,taurine and hypotaurine metabolism,and biosynthesis of unsaturated fatty acids were co-enriched in differential proteins and differential metabolites,involving three differential proteins and nine differential metabolites.Among these,docosapentaenoic acid(22n-3),eicosapentaenoic acid,taurine,3-sulfinoalanine,taurocholic acid,Acnat1,Acnat2,and Hsd17b12 showed high prediction accuracy.Conclusions We successfully constructed an RF animal model of Qi deficiency and dampness stasis using the "cyclosporine A+high-fat diet+exhaustion of swimming+constant temperature and humidity" method.Biosynthesis of unsaturated fatty acids and taurine and hypotaurine metabolism may play important roles in this RF mouse model of Qi deficiency and dampness stasis.

Objective To construct and evaluate a mouse model of renal fibrosis(RF)combined with Qi deficiency and dampness stasis,and investigate the changes in protein and metabolic pathways using multiomics.Methods Twenty-four C57BL/6J mice were divided randomly into normal(N),model(M),and RF and syndrome combined groups(BZ)(n=8/group).The experiment lasted 6 weeks.A mouse model of RF with Qi deficiency and dampness stasis was established by "cyclosporine A+high-fat diet+swimming exhaustion+constant temperature and humidity".The model was evaluated by detecting general signs,renal function,tongue RGB(red,green,blue)values,hemorheology indexes,blood lipids,and inflammation and oxidation indexes,combined with hematoxylin and eosin,Masson,periodic acid-Schiff,and Oil red O staining,terminal deoxynucleotidyl transferase dUTP nick end labeling apoptosis,and transforming growth factor-β immunofluorescence analysis of renal tissue.Differential proteins and metabolites were screened by renal proteomics combined with serum metabolomics and subjected to pathway enrichment analysis.Results Body mass of mice in the BZ group began to decline at week 3(P＜0.05)and decreased significantly at week 4(P＜0.01),while food and water consumption decreased,the fur became messy and less glossy,mood and activity decreased,and stools became watery.Serum creatinine,blood urea nitrogen,urine albumin-creatinine ratio,and N-acetyl-beta-glucosaminidase(NAG)were significantly higher in the BZ group compared with those in the N group(P＜0.05,P＜0.01),and serum creatinine and NAG levels were significantly different compared with those in the M group.The R value of tongue images was significantly lower in the BZ group compared with that in the N group(P＜0.01),while the B value was significantly higher(P＜0.05).The viscosity of the whole blood multi-shear rate and the hematocrit were higher in the BZ group compared with those in the N and M groups,and the platelet volume was higher than in the N group(P＜0.05,P＜0.01).Total cholesterol,low-density lipoprotein cholesterol,C-reactive protein,interleukin-6,and malondialdehyde levels were significantly increased in the BZ group compared with those in the N and M groups(P＜0.01),and superoxide dismutase activity was significantly decreased compared with that in the N group(P＜0.05).Renal tubule vacuolation,inflammatory cell infiltration,glomerular basement membrane thickening,collagen fiber hyperplasia,and lipid accumulation were evident,and renal cell apoptosis and transforming growth factor-β deposition were increased in the BZ group.There were 299 differential proteins in the BZ and N groups,including 180 up-regulated and 119 down-regulated proteins,and 323 differential metabolites,including 205 up-regulated and 118 down-regulated.Primary bile acid biosynthesis,taurine and hypotaurine metabolism,and biosynthesis of unsaturated fatty acids were co-enriched in differential proteins and differential metabolites,involving three differential proteins and nine differential metabolites.Among these,docosapentaenoic acid(22n-3),eicosapentaenoic acid,taurine,3-sulfinoalanine,taurocholic acid,Acnat1,Acnat2,and Hsd17b12 showed high prediction accuracy.Conclusions We successfully constructed an RF animal model of Qi deficiency and dampness stasis using the "cyclosporine A+high-fat diet+exhaustion of swimming+constant temperature and humidity" method.Biosynthesis of unsaturated fatty acids and taurine and hypotaurine metabolism may play important roles in this RF mouse model of Qi deficiency and dampness stasis.

Objective To explore the factors influencing the blood concentration of voriconazole in elderly hospitalized patients and inform the probability of attaining the target concentration in clinical practice.Methods Patients aged ≥65 years who were hospitalized in the First Affiliated Hospital of Anhui Medical University from January 2022 to December 2023 and underwent voriconazole blood concentration monitoring were enrolled.Their voriconazole blood concentrations and clinical data were collected.The patients were grouped according to the target effective concentration 0.5-5.0 mg/L of voriconazole recommended by the Chinese Pharmacological Society guidelines.Multivariate logistic regression analysis was used to determine the factors affecting the rate of achieving the target concentration.Results The 202 enrolled patients included 139 males and 63 females.A total of 244 voriconazole blood concentrations were available.The median age of the patients was 74(range:65-95)years.Voriconazole blood concentration ranged from 0.08 to 13.38 mg/L.The average concentration was(4.10±2.45)mg/L.The target effective blood concentration of voriconazole was achieved in 65.35％(132/202)of the patients.Logistic regression results showed that the dosage regimen,body weight,and hypoproteinemia(albumin＜25 g/L)were the main factors affecting voriconazole blood concentration.Conclusions The dosing regimen,body weight,and hypoproteinemia are the main influencing factors of voriconazole blood concentration.Relevant factors should be fully considered in clinical medication to ensure the safety and effectiveness of voriconazole.

Objective To explore the factors influencing the blood concentration of voriconazole in elderly hospitalized patients and inform the probability of attaining the target concentration in clinical practice.Methods Patients aged ≥65 years who were hospitalized in the First Affiliated Hospital of Anhui Medical University from January 2022 to December 2023 and underwent voriconazole blood concentration monitoring were enrolled.Their voriconazole blood concentrations and clinical data were collected.The patients were grouped according to the target effective concentration 0.5-5.0 mg/L of voriconazole recommended by the Chinese Pharmacological Society guidelines.Multivariate logistic regression analysis was used to determine the factors affecting the rate of achieving the target concentration.Results The 202 enrolled patients included 139 males and 63 females.A total of 244 voriconazole blood concentrations were available.The median age of the patients was 74(range:65-95)years.Voriconazole blood concentration ranged from 0.08 to 13.38 mg/L.The average concentration was(4.10±2.45)mg/L.The target effective blood concentration of voriconazole was achieved in 65.35％(132/202)of the patients.Logistic regression results showed that the dosage regimen,body weight,and hypoproteinemia(albumin＜25 g/L)were the main factors affecting voriconazole blood concentration.Conclusions The dosing regimen,body weight,and hypoproteinemia are the main influencing factors of voriconazole blood concentration.Relevant factors should be fully considered in clinical medication to ensure the safety and effectiveness of voriconazole.

Background: Nasopharyngeal carcinoma (NPC) is characterized by high programmed death-ligand 1 (PD-L1) expression and abundant infiltration of non-malignant lymphocytes, which renders patients potentially suitable candidates for immune checkpoint blockade therapies. Palate, lung, and nasal epithelium clone (PLUNC) inhibit the growth of NPC cells and enhance cellular apoptosis and differentiation. Currently, the relationship between PLUNC (as a tumor-suppressor) and PD-L1 in NPC is unclear. Methods: We collected clinical samples of NPC to verify the relationship between PLUNC and PD-L1. PLUNC plasmid was transfected into NPC cells, and the variation of PD-L1 was verified by western blot and immunofluorescence. In NPC cells, we verified the relationship of PD-L1, activating transcription factor 3 (ATF3), and β-catenin by western blot and immunofluorescence. Later, we further verified that PLUNC regulates PD-L1 through β-catenin. Finally, the effect of PLUNC on β-catenin was verified by co-immunoprecipitation (Co-IP). Results: We found that PLUNC expression was lower in NPC tissues than in paracancer tissues. PD-L1 expression was opposite to that of PLUNC. Western blot and immunofluorescence showed that β-catenin could upregulate ATF3 and PD-L1, while PLUNC could downregulate ATF3/PD-L1 by inhibiting the expression of β-catenin. PLUNC inhibits the entry of β-catenin into the nucleus. Co-IP experiments demonstrated that PLUNC inhibited the interaction of DEAD-box helicase 17 (DDX17) and β-catenin. Conclusions PLUNC downregulates the expression of PD-L1 by inhibiting the interaction of DDX17/β-catenin in NPC.

Background: Nasopharyngeal carcinoma (NPC) is characterized by high programmed death-ligand 1 (PD-L1) expression and abundant infiltration of non-malignant lymphocytes, which renders patients potentially suitable candidates for immune checkpoint blockade therapies. Palate, lung, and nasal epithelium clone (PLUNC) inhibit the growth of NPC cells and enhance cellular apoptosis and differentiation. Currently, the relationship between PLUNC (as a tumor-suppressor) and PD-L1 in NPC is unclear. Methods: We collected clinical samples of NPC to verify the relationship between PLUNC and PD-L1. PLUNC plasmid was transfected into NPC cells, and the variation of PD-L1 was verified by western blot and immunofluorescence. In NPC cells, we verified the relationship of PD-L1, activating transcription factor 3 (ATF3), and β-catenin by western blot and immunofluorescence. Later, we further verified that PLUNC regulates PD-L1 through β-catenin. Finally, the effect of PLUNC on β-catenin was verified by co-immunoprecipitation (Co-IP). Results: We found that PLUNC expression was lower in NPC tissues than in paracancer tissues. PD-L1 expression was opposite to that of PLUNC. Western blot and immunofluorescence showed that β-catenin could upregulate ATF3 and PD-L1, while PLUNC could downregulate ATF3/PD-L1 by inhibiting the expression of β-catenin. PLUNC inhibits the entry of β-catenin into the nucleus. Co-IP experiments demonstrated that PLUNC inhibited the interaction of DEAD-box helicase 17 (DDX17) and β-catenin. Conclusions PLUNC downregulates the expression of PD-L1 by inhibiting the interaction of DDX17/β-catenin in NPC.

Background: Nasopharyngeal carcinoma (NPC) is characterized by high programmed death-ligand 1 (PD-L1) expression and abundant infiltration of non-malignant lymphocytes, which renders patients potentially suitable candidates for immune checkpoint blockade therapies. Palate, lung, and nasal epithelium clone (PLUNC) inhibit the growth of NPC cells and enhance cellular apoptosis and differentiation. Currently, the relationship between PLUNC (as a tumor-suppressor) and PD-L1 in NPC is unclear. Methods: We collected clinical samples of NPC to verify the relationship between PLUNC and PD-L1. PLUNC plasmid was transfected into NPC cells, and the variation of PD-L1 was verified by western blot and immunofluorescence. In NPC cells, we verified the relationship of PD-L1, activating transcription factor 3 (ATF3), and β-catenin by western blot and immunofluorescence. Later, we further verified that PLUNC regulates PD-L1 through β-catenin. Finally, the effect of PLUNC on β-catenin was verified by co-immunoprecipitation (Co-IP). Results: We found that PLUNC expression was lower in NPC tissues than in paracancer tissues. PD-L1 expression was opposite to that of PLUNC. Western blot and immunofluorescence showed that β-catenin could upregulate ATF3 and PD-L1, while PLUNC could downregulate ATF3/PD-L1 by inhibiting the expression of β-catenin. PLUNC inhibits the entry of β-catenin into the nucleus. Co-IP experiments demonstrated that PLUNC inhibited the interaction of DEAD-box helicase 17 (DDX17) and β-catenin. Conclusions PLUNC downregulates the expression of PD-L1 by inhibiting the interaction of DDX17/β-catenin in NPC.

OBJECTIVE: Benzylisoquinoline alkaloids (BIAs) have pharmacological functions and clinical use. BIAs are mainly distributed in plant species across the order Ranunculales and the genus Phellodendron from Sapindales. The BIA biosynthesis has been intensively investigated in Ranunculales species. However, the accumulation mechanism of BIAs in Phellodendron is largely unknown. The aim of this study is to unravel the biosynthetic pathways of BIAs in Phellodendron amurens. METHODS: The transcriptome and metabolome data from 18 different tissues of P. amurense were meticulously sequenced and subsequently subjected to a thorough analysis. Weighted gene co-expression network analysis (WGCNA), a powerful systems biology approach that facilitates the construction and subsequent analysis of co-expression networks, was utilized to identify candidate genes involved in BIAs biosynthesis. Following this, recombinant plasmids containing candidate genes were expressed in Escherichia coli, a widely used prokaryotic expression system. The purpose of this genetic engineering endeavor was to express the candidate genes within the bacteria, thereby enabling the assessment of the resultant enzyme activity. RESULTS: The synonymous substitutions per synonymous site for paralogs indicated that at least one whole genome duplication event has occurred. The potential BIA biosynthetic pathway of P. amurense was proposed, and two PR10/Bet v1 members, 14 CYP450s, and 33 methyltransferases were selected as related to BIA biosynthesis. One PR10/Bet v1 was identified as norcoclaurine synthase, which could catalyze dopamine and 4-hydroxyphenylacetaldehyde into (S)-norcoclaurine. CONCLUSION Our studies provide important insights into the biosynthesis and evolution of BIAs in non-Ranunculales species.

Objective To explore the prescription rules of traditional Chinese medicine (TCM) in the treatment of diabetes periodontitis(DP) and the acting mechanisms of core drug combination. Methods Based on the relevant literature retrieved from the CNKI,Wanfang,VIP and Sinomed,a DP prescription database was established. Excel 2021,SPSS Modeler 18.0 and SPSS Statistics 26.0 were used to conduct the statistics of the frequency,efficacy classifications,properties,flavors,and meridian tropism of the included drugs. Association rule analysis and cluster analysis were performed to screen out the core drug combinations. The active components and action targets of core drug combinations were obtained through TCMSP and HERB. The DP related disease targets were predicted using GeneCards. The Venny platform was used to obtain the intersection of disease targets and drug targets. Key components were screened by Cytoscape to establish an "active component-target" network. Based on STRING platform data,PPI network was constructed by Cytoscape to screen core targets. GO functional annotation and KEGG signaling pathway enrichment analysis were carried out for the intersection targets by DAVID. AutoDockVina was applied for molecular docking between core targets and key components. Results A total of 36 articles were included,and 50 prescriptions involving 100 Chinese herbal medicines were extracted. Alismatis Rhizoma,Rehmanniae Radix Praeparata and Astragali Radix were the most common drugs. The most used drug category was deficiency-nourishing drugs. The properties of the herbs were mainly cold and warm,the major flavors were sweet and bitter,and the main meridian tropisms were kidney and liver. Six categories were classified by clustering analysis. Moutan Cortes-Corni Fructus-Rehmanniae Radix Praeparata was screened out as the core drug combination involving 18 active components,164 drug action targets and 104 intersection of DP targets and drug combination targets. Quercetin,stigmasterol,kaempferol,β-sitosterol,tetrahydroalstonine,and sitosterol were the key components,and AKT1,IL-6,TNF,IL-1B,PTGS2,JUN,TP53,ESR1,and MMP9 were the core targets. GO analysis revealed 3724 biological processes,228 cellular components and 404 molecular functions. KEGG analysis showed that DP was treated by the core drug combination through regulating 235 signaling pathways. Molecular docking results showed that there was a good affinity between the core target and the key component. Conclusion Tonifying deficiency is the main treatment methods of TCM for DP,accompanied by clearing heat and removing dampness,activating blood circulation and removing blood stasis,replenishing qi and nourishing yin. Core drug combination (Moutan Cortes-Corni Fructus-Rehmanniae Radix Praeparata) treats DP through multi-component,multi-target and multi-pathway,which provide a reference for clinical diagnosis and treatment.

Renal fibrosis， the final pathological outcome of end-stage chronic kidney diseases， is associated with inflammation， oxidative stress， epithelial-mesenchymal transdifferentiation （EMT）， and extracellular matrix deposition. It belongs to the categories of edema， ischuria， anuria and vomiting， and consumptive disease in traditional Chinese medicine （TCM）， with the key pathogenesis of Qi deficiency and blood stasis and the primary treatment principle of replenishing Qi and activating blood. Astragali Radix-Salviae Miltiorrhizae Radix et Rhizoma mainly contains astragalosides， polysaccharides， calycosin， salvianolic acid， and tanshinone， with the effect of tonifying Qi and activating blood. Studies have shown that this herb pair and its active components can delay the progress of renal fibrosis by regulating multiple signaling pathways. With consideration to the pathogenesis of Qi deficiency and blood stasis， this article reviews the research progress in the mitigation of renal fibrosis by Astragali Radix-Salviae Miltiorrhizae Radix et Rhizoma from the aspects of protecting glomerular filtration barrier， inhibiting EMT and mesangial cell proliferation， improving renal hemodynamics， and protecting renal function. Furthermore， the mechanisms were summarized. Specifically， Astragali Radix-Salviae Miltiorrhizae Radix et Rhizoma and its effective components can improve mitochondrial function and fatty acid metabolism， alleviate endoplasmic reticulum stress and autophagy disorders， and inhibit immune inflammation and oxidative stress by regulating nuclear factor E₂-related factor 2 （Nrf2）/PTEN-induced kinase 1 （Pink1）， Nrf2/antioxidant response element （ARE）， tumor necrosis factor-α （TNF-α）/nuclear transcription factor-κB （NF-κB）， miR-21/Smad7/transforming growth factor beta （TGF-β）， Wnt/β-catenin， long non-coding RNA-taurine up-regulated gene 1 （lncRNA-TUG1）/tumor necrosis factor receptor-associated factor 5 （TRAF5）， Ras-related C3 botulinum toxin substrate 1 （Rac1）/cell division cycle protein 42 （CDC42）， Ras homolog （Rho）/Rho-associated coiled-coil containing protein kinase （ROCK）， phosphatidylinositol-3-kinase （PI3K）/protein kinase B （Akt）， Janus kinase （JAK）/signal transducer and activator of transcription （STAT）， peroxisome proliferator-activated receptor α （PPARα）/peroxisome proliferator-activated receptor γ coactivator l alpha （PGC-1α）， and p38 mitogen-activated protein kinase （p38 MAPK）. This review aims to provide references for the relevant research， give play to the role of Astragali Radix-Salviae Miltiorrhizae Radix et Rhizoma， and provide guidance for the clinical treatment of renal fibrosis.