Objective To elucidate the molecular mechanism of sirtuin-3 (SIRT3) in regulating mitochondrial biogenesis in human renal tubular epithelial cells. Methods Cells were stimulated with different concentrations of H₂O₂ and divided into four groups: control (NC), 50 μmol/L H₂O₂, 110 μmol/L H₂O₂ and 150 μmol/L H₂O₂. SIRT3 protein expression was then measured. SIRT3 was knocked down with siRNA, and cells were further assigned to five groups: control (NC), negative-control siRNA (NCsi), SIRT3-siRNA (siSIRT3), NCsi+H₂O₂, and siSIRT3+H₂O₂. After 24 h, cellular adenosine triphosphate (ATP) and mitochondrial superoxide anion (O₂•⁻) levels were determined, together with mitochondrial expression of SIRT3, peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1α), nuclear respiratory factor 1 (NRF1), mitochondrial transcription factor A (TFAM), superoxide dismutase 2 (SOD2), acetylated-SOD2 and adenosine monophosphate activated protein kinase α1 (AMPKα1). Results The 110 and 150 μmol/L H₂O₂ decreased SIRT3 protein (both P<0.05). ATP and mitochondrial O₂•⁻ did not differ between NC and NCsi groups (both P>0.05). Compared to the NCsi group, the siSIRT3 group exhibited elevated O₂•⁻ level, decreased SIRT3 protein and increased expression levels of SOD2 and acetylated SOD2 protein (all P<0.05). Compared to the NCsi group, the NCsi+H₂O₂ group exhibited decreased cellular ATP levels, elevated mitochondrial O₂•⁻ levels, and reduced protein expression levels of SIRT3, SOD2, TFAM, AMPKα1, PGC-1α and NRF1 (all P<0.05). Compared with the siSIRT3 group, the siSIRT3+H₂O₂ group showed a decrease in cellular ATP levels, an increase in mitochondrial O₂•⁻ levels, a decrease in SIRT3, SOD2, TFAM, AMPKα1, PGC-1α and NRF1 protein expression levels and a decrease in acetylated SOD2 protein expression levels (all P<0.05). Compared with the NCsi+H₂O₂ group, the siSIRT3+H₂O₂ group showed a decrease in cellular ATP levels, an increase in mitochondrial O₂•⁻ levels, a decrease in SIRT3, AMPKα1, PGC-1α and NRF1, TFAM protein expression levels, and an increase in SOD2 and acetylated SOD2 protein expression levels (all P<0.05). Conclusions SIRT3 promotes mitochondrial biogenesis in tubular epithelial cells via the AMPK/PGC-1α/NRF1/TFAM axis, representing a key mechanism through which SIRT3 ameliorates oxidative stress-induced mitochondrial dysfunction.

To introduce the general thinking, guidelines, work objectives and elaboration process of the general technical requirements adopted by volume Ⅳ of the Chinese Pharmacopoeia 2025 Edition, and to summarize and figure out the main characteristics on dosage forms, physico-chemical testing, microbial and biological testing, reference standards and guidelines The newly revised general chapters of pharmacopoeia give full play to the normative and guiding role of the Chinese Pharmacopoeia standard, track the frontier dynamics of international drug regulatory science and the elaboration of monographs, expand the application of state-of-the-art technologies, and steadily promote the harmonization and unification with the ICH guidelines； further enhance the overall capacity of TCM quality control, actively implement the 3 R principles on animal experiments, and practice the concept of environmental-friendly； replace and/or reduce the use of toxic and hazardous reagents, strengthen the requirements of drug safety control This paper aims to provide a full-view perspective for the comprehensive, correct understanding and accurate implementation of general technical requirements included in the Chinese Pharmacopoeia 2025 Edition.

Objective To evaluate the diagnostic performance of the minimum-cost-path-based CT angiography-derived fractional flow reserve(MCP-FFR)and the deep learning-driven CT angiography-derived fractional flow reserve(DeepCL-FFR),and to particularly explore the potential value of the DeepCL algorithm in improving diagnostic accuracy within the"gray zone."Methods A retrospective analysis was conducted on 151 coronary vessels from 109 patients with coronary artery disease,who were hospitalized at the General Hospital of the People's Liberation Army between January 2020 and June 2021.Pearson correlation and Bland-Altman plots were employed to assess the correlation and agreement of the two CT-FFR methods with invasive FFR.A CT-FFR range of 0.70-0.80 was defined as the diagnostic"gray zone."The accuracy,sensitivity,specificity,positive predictive value,and negative predictive value for detecting hemodynamic abnormalities were calculated and analyzed.The DeLong test was used to compare the areas under the receiver operating characteristic curves(AUC)between the two CT-FFR calculation methods.Results Both CT-FFR methods exhibited a positive correlation with invasive FFR(MCP-FFR:r=0.75,P＜0.001;DeepCL-FFR:r=0.86,P＜0.001)and showed good agreement(MCP-FFR:mean difference=0.010,P=0.351;DeepCL-FFR:mean difference=-0.003,P=0.772).Both DeepCL-FFR(AUC 0.97,95％CI 0.94-0.99)and MCP-FFR(AUC 0.92,95％CI 0.88-0.97)demonstrated favorable diagnostic performance for detecting hemodynamic abnormalities(P=0.122).In the"gray zone"for hemodynamic abnormality,the diagnostic accuracy of MCP-FFR was 68.8％,whereas DeepCL-FFR increased it to 89.7％.DeepCL-FFR also exhibited superior diagnostic performance(AUC 0.89,95％CI 0.73-0.99)within the"gray zone,"which was significantly higher than that of MCP-FFR(AUC 0.71,95％CI 0.54-0.87)(P＜0.001).Conclusions The deep learning-driven coronary centerline extraction algorithm,DeepCL,demonstrates superior diagnostic performance in CT-FFR for detecting hemodynamic abnormalities,particularly by significantly improving diagnostic accuracy in the"gray zone."

Aim To investigate the pharmacological mechanism of Ebselenin(Ebselen,EbSe)in the treat-ment of Escherichia coli(E.coli)infection,which had no significant inhibitory effect on Gram-negative bacte-ria,based on previous studies.Methods After EbSe intervention in E.coli infected Raw264.7 cells,the via-bility of Raw264.7 cells was determined by CCK-8 method,the morphology and structure of Raw264.7 cells were observed by electron microscope,and the in-tracellular bacterial load of Raw264.7 cells was calcu-lated by coated plate method.Polarization status of peritoneal macrophages,Raw264.7 intracellular NO and ROS content and intracellular HO-1 expression in Raw264.7 and E.coli acutely infected mice after E.co-li infection by flow cytometry.qPCR was used to detect the expression of related mRNAs in Raw264.7 cells.qPCR was used to detect the intracellular GSH content in Raw264.7 cells by spectrophotometric assay,and the state of cytoskeletal proteins was observed by immuno-fluorescence.Western blot assay was performed to de-tect the intracellular Txnrd1 expression level.Results Microtiter method,CCK-8,and electron microscopy observations showed that EbSe had no effect on the growth of E.coli and Raw264.7 cells in vitro.The re-sults of smear plate counting showed that EbSe reduced the intracellular bacterial load of Raw264.7 in the in-fected group.Flow cytometry results showed that EbSe upregulated the number of M2-type macrophages.The EbSe-treated infected group had reduced intracellular NO and ROS levels and increased GSH levels.The qPCR results showed that the expression of IL-6,IL-1β,and iNOS was decreased,and the expression of HO-1,Txnrd1,and Glut1 was increased in DHB4-in-fected Raw264.7 cells after EbSe treatment.Cytoskel-etal staining showed that the morphology of the EbSe-treated infected cells was similar to that of oxPAPC-in-duced cells.Western blot results showed the expres-sion of Txnrd1 protein in EbSe-treated infected cells in-creased.Conclusion EbSe exerts anti-E.coli acute infection effect by regulating macrophage polarization and inhibiting macrophage excessive inflammatory state.

Aim To investigate the expression of or-ganic anion transporting polypeptide 4C1(Oatp4c1)-P-glycoprotein(P-gp)in the kidneys of obese mice in-duced by high-fat diet(HFD),and its impact on the pharmacokinetic changes of digoxin.Methods C57BL/6 mice were randomly divided into the Chow group and the HFD group.Body weight and blood glu-cose were recorded weekly.After successful model es-tablishment,digoxin was intraperitoneally injected,and blood was collected at different time points.Part of the blood samples was used for LC-MS/MS detection,and the other part was used for the detection of other bio-chemical indicators.After 16 weeks,the organs were removed and weighed.HE and immunohistochemical staining was used to observe the renal pathology and the expression of Villin,a marker of proximal tubules.Western blot and qPCR were combined to detect the expression of Villin,Oatp4c1 and P-gp.Results In the HFD group,body weight and blood glucose in-creased significantly.The blood concentration of digox-in rose,the area under the curve increased,and the half-life was prolonged.The proximal tubular epithelial cells shed,and the protein expression of Villin,Oatp4c1 and P-gp decreased significantly.Conclu-sions The down-regulation of Oatp4c1-P-gp expres-sion in the kidneys of HFD mice leads to an increase in the blood concentration of digoxin and a decrease in re-nal clearance.

Objective:Observation of the therapeutic effect of Yugu Ju detergent combined with fusidic acid cream on the repair of wounds infected with Staphylococcus aureus,as well as its impact on serum inflammatory markers TNF-α,IL-6,IL-1β,oxidative stress markers MDA,ROS,SOD and Nrf2 levels.Methods:96 patients with skin defects and SA U infection caused by trauma admitted to the hospital from June 2021 to December 2023 were randomly divided into a control group and an observation group.Both groups were treated with debridement,while the control group was treated with external application of fusidic acid cream.The treatment group received external washing with Yugu Ju detergent in addition to the control group.One course of treatment lasted for 7 days,with three consecutive courses of treatment.Observe the wound healing rate,bacterial clearance rate,and changes in TNF-α,IL-6,IL-1β,SOD,MDA,ROS,and Nrf2 before and after treatment to evaluate clinical efficacy and safety.Results:The total clinical effective rate of the observation group after treatment was 93.75％(45/48),while that of the control group was 75％(36/48).The observation group was significantly better than the control group(P＜0.05);The wound healing rate and bacterial clearance rate of the observation group at each time point after treatment were higher than those of the control group(P＜0.05);After treatment,the Nrf2 and SOD values in the observation group were higher than those in the control group,while the TNF-α,IL-6,IL-1β,MDA,and ROS values were lower than those in the control group(P＜0.05).No adverse reactions occurred during the treatment of both groups.Conclusion:The combination of Yugu Ju detergent and Fusidic acid cream can inhibit SA U and promote the healing of infectious wounds,which may be related to the activation of Nrf2 expression to inhibit oxidative stress and inflammatory response.It is safe and effective,and worthy of promotion and application.

Aim To study the effects of whole herb of Prunella and its active components on the malignant progression of breast cancer and its mechanism.Meth-ods Breast cancer transplantation tumor model was constructed and randomly divided into the model group,low,medium and high dose group of whole herb of Prunella(0.1,0.2,0.4 g·mL-1 by gavage)and paclitaxel(10 mg·kg-1 by intraperitoneal injection),which was administered by gavage every day,and the tumor tissues were collected after 28 days of interven-tion.The weight,tumor volume and mass of nude mice in each group were detected,HE staining was used to observe the morphology of breast cancer tumor tissues,and immunohistochemical staining was used to observe the proliferation of cell-cycle regulatory protein-67(Ki-67)and cytokeratin 17(CK17)in breast cancer tumor tissues.The cellular experiments were performed by u-sing different concentrations of the ethyl acetate extract of the whole herb of Prunella in breast cancer MDA-MB-231 cells for 24 h.The proliferation of MDA-MB-231 cells and the effects on the cell cycle and apoptosis of MDA-MB-231 cells were detected by using the CCK-8 assay,the cell cycle flow and the apoptotic cell flow.Western blot was used to detect the effect of ethyl ace-tate extract of whole herb of Prunella on the expression of apoptosis-related proteins in breast cancer MDA-MB-231 cells.UPLCQ-TOF MS/MS was used to detect the chemical compositions of the ethyl acetate extract of Prunella whole herb.Results The whole herb of Pru-nella had no significant effect on the growth of nude mice(P＞0.05);it could significantly inhibit the growth of transplanted tumors in nude mice with human breast cancer(P＜0.05);the results of HE staining showed that the cells in the tissues appeared to be rela-tively sparse with the increase of the dose of Prunella and had different degrees of nuclear consolidation and deep staining of nuclei and the apoptosis of the tumor cells increased;the metastasis of tumor cells to the liv-er and lungs was inhibited,when compared with that in the model group.Compared with the model group,the low,medium and high groups of Prunella had no signif-icant effect on the liver index,while the spleen index was significantly reduced(P＜0.05);the expression of Ki-67 and CK17 was reduced.The ethyl acetate ex-tract of the whole herb of Prunella could inhibit the proliferation of MDA-MB-231 cells in breast cancer(P＜0.01);the results of flow cytometry showed that,with the increase of the concentration of the ethyl ace-tate extract of the whole herb of Prunella,the proportion of S-phase cells in the MDA-MB-231 cells significantly increased,and the proportion of G0/G1-phase cells sig-nificantly decreased,while the proportion of G2-phase cells did not change significantly(P＜0.01);Western blotting was not affected in the low,medium and high groups,and the spleen index significantly decreased(P＜0.05);the expression of Ki-67 and CK17 was re-duced;the results of Western blot showed that the eth-yl acetate extract of the whole herb of Prunella promo-ted the expression of Bax,cleaved caspase-3,cleaved caspase-9 proteins,and inhibited the expression of Bcl-2,caspase-3,caspase-9,cyclinA2,and CDK2 proteins(P＜0.05,P＜0.01).The acetic acid of the whole herb of Prunella ethyl ester extract identified a total of 51 compounds.Conclusions The whole herb of Pru-nella can inhibit the growth of breast cancer in nude mice transplanted with tumors,promote the apoptosis of tumor cells,inhibit the proliferation of breast cancer MDA-MB-231 cells,inhibit the metastasis of tumor cells to the liver and lungs,protect the liver and spleen,and reduce the expression of the value-added markers Ki-67 and CK17 in tumor tissues,and the ef-fective ingredient of the whole herb,the ethyl acetate extract,can induce apoptosis.The mechanism may be related to the down-regulation of cyclinsA2,CDK2,Bcl-2,caspase-3,caspase-9 and up-regulation of Bax,cleaved caspase-3,cleaved caspase-9 protein expres-sion.

To introduce the general thinking,guidelines,work objectives and elaboration process of the general technical requirements adopted by volume Ⅳ of the Chinese Pharmacopoeia 2025 Edition,and to summarize and figure out the main characteristics on dosage forms,physico-chemical testing,microbial and biological testing,ref-erence standards and guidelines.The newly revised general chapters of pharmacopoeia give full play to the norma-tive and guiding role of the Chinese Pharmacopoeia standard,track the frontier dynamics of international drug regu-latory science and the elaboration of monographs,expand the application of state-of-the-art technologies,and stead-ily promote the harmonization and unification with the ICH guidelines;further enhance the overall capacity of TCM quality control,actively implement the 3 R principles on animal experiments,and practice the concept of environ-mental-friendly;replace and/orreduce the use of toxic and hazardousreagents,strengthen the requirementsofdrug safety control.This paper aims to provide a full-view perspective for the comprehensive,correct understanding and accurate implementation of general technical requirements included in the Chinese Pharmacopoeia 2025 Edition.

Objective:To systematically analyze the revisions content and technological development trends of microbiological standards in the Chinese Pharmacopoeia(ChP)2025 Edition,and explore its novel requirements in risk-based pharmaceutical product lifecycle management.Methods:A comprehensive review was conducted on 26 microbiological-related standards to summarize the revision directions and scientific implications from perspectives including the revision overview,international harmonization of microbiological standards,risk-based quality man-agement system,and novel tools and methods with Chinese characteristics.Results:The ChP 2025 edition demon-strates three prominent features in microbiological-related standards:enhanced international harmonization,intro-duced emerging molecular biological technologies,and established a risk-based microbiological quality control sys-tem.Conclusion:The new edition of the Pharmacopoeia has systematically constructed a microbiological standard system,which significantly improves the scientificity,standardization and applicability of the standards,providing a crucial support for advancing the microbiological quality control in pharmaceutical industries of China.

Objective To analyze the global literature related to functional cure of hepatitis B from 2019 to 2023 by using bibliometric analysis methods,so as to help researchers understand the research hotspots and trends in this field.Methods The literature related to the topic of functional cure of hepatitis B included in the Science Citation Index Expanded(SCI-Expanded)of the Web of Science Core Collection from 2019 to 2023 was searched.By using VOSviewer and CiteSpace visual analysis tools,analyses were conducted from the perspectives of publication trends,international research cooperation networks,and keyword emergence,and were elaborated with the specific contents of the related literature to elucidate research hotspots and trends.Results A total of 600 eligible papers in this field were included.Keyword co-occurrence and thematic clustering suggested that the main research directions of functional cure were:serum biomarkers for prediction and monitoring of functional cure,functional cure and immunity,nucleoside analog discontinuation,interferon therapy,and long-term prognosis of functional cure.The research contents of the ESI highly cited original research papers were similar to the clustering of the above,but showed more attention on the novel agents for functional cure.The content of the keyword emergence map showed that hotspots of interest changed from virologic mechanisms and serum markers,to nucleoside analog discontinuation and interferon therapy,and finally to immunologic mechanisms and new drug.Conclusion The research hotspots and trends of functional cure of hepatitis B were focused on virological mechanism,serum markers,immunological mechanism,nucleoside analog discontinuation,interferon therapy,and long-term prognosis after cure.