Animal experiments are widely used in biomedical research for safety assessment, toxicological analysis, efficacy evaluation, and mechanism exploration. In recent years, the ethical review system has become more stringent, and awareness of animal welfare has continuously increased. To promote more efficient and cost-effective drug research and development, the United States passed the Food and Drug Administration (FDA) Modernization Act 2.0 in September 2022, which removed the federal mandate requiring animal testing in preclinical drug research. In April 2025, the FDA further proposed to adopt a series of "new alternative methods" in the research and development of drugs such as monoclonal antibodies, which included artificial intelligence computing models, organoid toxicity tests, and 3D micro-physiological systems, thereby gradually phasing out traditional animal experiment models. Among these cutting-edge technologies, 3D bioprinting models are a significant alternative and complement to animal models, owing to their high biomimetic properties, reproducibility, and scalability. This review provides a comprehensive overview of advancements and applications of 3D bioprinting technology in the fields of biomedical and pharmaceutical research. It starts by detailing the essential elements of 3D bioprinting, including the selection and functional design of biomaterials, along with an explanation of the principles and characteristics of various printing strategies, highlighting the advantages in constructing complex multicellular spatial structures, regulating microenvironments, and guiding cell fate. It then discusses the typical applications of 3D bioprinting in drug research and development,including high-throughput screening of drug efficacy by constructing disease models such as tumors, infectious diseases, and rare diseases, as well as conducting drug toxicology research by building organ-specific models such as those of liver and heart. Additionally,the review examines the role of 3D bioprinting in tissue engineering, discussing its contributions to the construction of functional tissues such as bone, cartilage, skin, and blood vessels, as well as the latest progress in regeneration and replacement. Furthermore, this review analyzes the complementary advantages of 3D bioprinting models and animal models in the research of disease progression, drug mechanisms, precision medicine, drug development, and tissue regeneration, and discusses the potential and challenges of their integration in improving model accuracy and physiological relevance. In conclusion, as a cutting-edge in vitro modeling and manufacturing technology, 3D bioprinting is gradually establishing a comprehensive application system covering disease modeling, drug screening, toxicity prediction, and tissue regeneration.

Animal experiments are widely used in biomedical research for safety assessment, toxicological analysis, efficacy evaluation, and mechanism exploration. In recent years, the ethical review system has become more stringent, and awareness of animal welfare has continuously increased. To promote more efficient and cost-effective drug research and development, the United States passed the Food and Drug Administration (FDA) Modernization Act 2.0 in September 2022, which removed the federal mandate requiring animal testing in preclinical drug research. In April 2025, the FDA further proposed to adopt a series of "new alternative methods" in the research and development of drugs such as monoclonal antibodies, which included artificial intelligence computing models, organoid toxicity tests, and 3D micro-physiological systems, thereby gradually phasing out traditional animal experiment models. Among these cutting-edge technologies, 3D bioprinting models are a significant alternative and complement to animal models, owing to their high biomimetic properties, reproducibility, and scalability. This review provides a comprehensive overview of advancements and applications of 3D bioprinting technology in the fields of biomedical and pharmaceutical research. It starts by detailing the essential elements of 3D bioprinting, including the selection and functional design of biomaterials, along with an explanation of the principles and characteristics of various printing strategies, highlighting the advantages in constructing complex multicellular spatial structures, regulating microenvironments, and guiding cell fate. It then discusses the typical applications of 3D bioprinting in drug research and development,including high-throughput screening of drug efficacy by constructing disease models such as tumors, infectious diseases, and rare diseases, as well as conducting drug toxicology research by building organ-specific models such as those of liver and heart. Additionally,the review examines the role of 3D bioprinting in tissue engineering, discussing its contributions to the construction of functional tissues such as bone, cartilage, skin, and blood vessels, as well as the latest progress in regeneration and replacement. Furthermore, this review analyzes the complementary advantages of 3D bioprinting models and animal models in the research of disease progression, drug mechanisms, precision medicine, drug development, and tissue regeneration, and discusses the potential and challenges of their integration in improving model accuracy and physiological relevance. In conclusion, as a cutting-edge in vitro modeling and manufacturing technology, 3D bioprinting is gradually establishing a comprehensive application system covering disease modeling, drug screening, toxicity prediction, and tissue regeneration.

Objective:Paragangliomas （PGLs） are chromaffin cell tumors originating from paraganglia and are classified as neuroendocrine neoplasms.They predominantly occur along the distribution area of the paraganglia, commonly occurring between the ages of 20 and 40, with a slight male predominance.They are most frequently found in the axial regions from the skull base to the pelvic cavity. Paragangliomas in the head and neck region typically lack endocrine functionality and primarily manifest through local mass effects. However, clinical signs and symptoms alone cannot reliably distinguish between metastatic and non-metastatic cases. Clinically apparent metastatic paragangliomas are relatively rare. Herein, we present a case of a paraganglioma located in the region of the jugular foramen with liver, bone, and lymph node metastases, and discuss the treatment and prognosis of head and neck paragangliomas.
Humans ; Head and Neck Neoplasms/pathology* ; Jugular Foramina/pathology* ; Liver Neoplasms/secondary* ; Lymphatic Metastasis ; Paraganglioma/pathology*

Deactivation of the mitochondrial pyruvate dehydrogenase complex (PDC) is important for the metabolic switching of cancer cell from oxidative phosphorylation to aerobic glycolysis. Studies examining PDC activity regulation have mainly focused on the phosphorylation of pyruvate dehydrogenase (E1), leaving other post-translational modifications largely unexplored. Here, we demonstrate that the acetylation of Lys 488 of pyruvate dehydrogenase complex component X (PDHX) commonly occurs in hepatocellular carcinoma, disrupting PDC assembly and contributing to lactate-driven epigenetic control of gene expression. PDHX, an E3-binding protein in the PDC, is acetylated by the p300 at Lys 488, impeding the interaction between PDHX and dihydrolipoyl transacetylase (E2), thereby disrupting PDC assembly to inhibit its activation. PDC disruption results in the conversion of most glucose to lactate, contributing to the aerobic glycolysis and H3K56 lactylation-mediated gene expression, facilitating tumor progression. These findings highlight a previously unrecognized role of PDHX acetylation in regulating PDC assembly and activity, linking PDHX Lys 488 acetylation and histone lactylation during hepatocellular carcinoma progression and providing a potential biomarker and therapeutic target for further development.
Humans ; Acetylation ; Carcinoma, Hepatocellular/genetics* ; Liver Neoplasms/genetics* ; Pyruvate Dehydrogenase Complex/genetics* ; Gene Expression Regulation, Neoplastic ; Animals ; Mice ; Cell Line, Tumor ; Protein Processing, Post-Translational ; Histones/metabolism* ; Disease Progression

BACKGROUND:Circular RNAs(circRNAs)play important roles in a variety of diseases or tumors,and recent findings have revealed that circRNAs are abnormally expressed in knee osteoarthritis and are involved in disease progression through microRNA/mRNA regulation. OBJECTIVE:To investigate the effect of circRNA WD repeat containing protein 1(circ-BRWD1)/miR-488-3p/DNA methyltransferase 3A(DNMT3A)on the proliferation and apoptosis of chondrocytes in knee osteoarthritis. METHODS:Real-time fluorescence quantitative PCR was performed to detect the expression of circ-BRWD1,miR-488-3p,DNMT3A in knee osteoarthritis chondrocytes.Cells were divided into si-NC group,si-circ-BRWD1 group,vector group,circ-BRWD1 group,si-circ-BRWD1+anti-miR-NC group,si-circ-BRWD1+anti-miR-488-3p group,miR-NC group,miR-488-3p group,anti-miR-NC group,anti-miR-488-3p group,miR-488-3p+vector group,miR 488-3p+DNMT3A group.Real-time fluorescence quantitative PCR was used to detect circ-BRWD1,miR-488-3p,DNMT3A expression,MTT and flow cytometry assay were used to detect cell proliferation and apoptosis.Western blot assay was used to detect DNMT3A and proliferation/apoptosis-related protein expression.Dual luciferase reporter assay was used to Dual luciferase reporter assay to detect the targeting relationship of circ-BRWD1 with miR-488-3p and miR-488-3p with DNMT3A. RESULTS AND CONCLUSION:circ-BRWD1 and DNMT3A were highly expressed and miR-488-3p was lowly expressed in knee osteoarthritis chondrocytes compared with normal chondrocytes.Knockdown of circ-BRWD1 or overexpression of miR-488-3p inhibited proliferation and induced apoptosis in knee osteoarthritis chondrocytes.circ-BRWD1 targeted negative regulation of miR-488-3p and inhibition of miR-488-3p reversed the effect of circ-BRWD1 knockdown on chondrocyte proliferation and apoptosis in knee osteoarthritis.miR-488-3p targeted negative regulation of DNMT3A and upregulation of DNMT3A reversed the effect of miR-488-3p overexpression on chondrocyte proliferation and apoptosis in knee osteoarthritis.circ-BRWD1 could regulate the expression of DNMT3A by regulating miR-488-3p.To conclude,knockdown of circ-BRWD1 inhibits chondrocyte proliferation and induces apoptosis in knee osteoarthritis,and the mechanism of action may be related to the regulation of miR-488-3p/DNMT3A axis.

OBJECTIVE To prepare patchouli oil enteric-coated dropping pills, evaluate its colon-targeted release behaviors and therapeutic potency against rat ulcerative colitis(UC). METHODS The single factor combined with response surface optimization method was used to screen matrix types and optimize preparation process parameters. Formula and thickness of Eudragit coating was selected based on dissolution tendency toward simulated intestinal fluids. Finally, colon targeting release behavior and the therapeutic effect of the preparation were assessed on the rat UC model induced by 2,4,6-trinitrobenzene sulfonic acid(TNBS). RESULTS The optimal prescription of patchouli oil dropping pills was patchouli oil∶PEG6000∶PEG8000 ratio of 1∶1∶1; and the optimal condition for preparing patchouli oil pills was keeping nozzle temperature at 9 ℃, and dropping pills at the speed of 33 drops·min−1, with dropping distance set at 6 cm; the optimal ratio of Eudragit L100∶Eudragit S100 was 3∶7 for preferential release in simulate intestinal fluid over simulated gastric fluid. Compared with free patchouli oil, patchouli oil enteric-coated dropping pills significantly alleviated the pathological symptoms such as weight loss, hematochezia and colon shortening in rats; the expression of pro-inflammatory cytokines IL-6, IL-1β, and IL-23 in serum was significantly down-regulated and the expression of anti-inflammatory cytokines IL-10 and TGF-β1 was significantly up-regulated. The mRNA expression of Mucin-1 and Mucin-2 in colon tissue was significantly up-regulated and the mRNA expression of inflammatory cytokines IL-6, IL-1β, and TNF-α was significantly down-regulated. CONCLUSION The patchouli oil enteric-coated dropping pills have colon-targeted release ability and improve the anti-inflammatory effect of drugs.

Objective:To evaluate the efficacy of electroacupuncture(EA)in enhancing the recovery of gastrointestinal function after laparoscopic cholecystectomy(LC). Methods:Randomized controlled trials(RCTs)of EA treatment in the postoperative period of patients undergoing LC were searched.Studies were obtained from Excerpta Medica Database(EMBASE),PubMed,Cochrane Library,Wanfang Academic Journal Full-text Database(Wanfang),China National Knowledge Infrastructure(CNKI),China Biology Medicine Disc(CBM),and Chongqing VIP Database(CQVIP)from inception to December 10th,2022.RevMan 5.4.1 was used to perform the meta-analysis.The Cochrane tool was used to assess the risk of bias.Mean difference(MD)and confidence interval(CI)were used for statistical descriptions. Results:A total of 7 studies were included in the meta-analysis.The meta-analysis found that the EA group had a shorter time to the first flatus[P<0.001,MD=-5.32,95%CI(-6.42,-4.21)],bowel movement recovery[P<0.001,MD=-6.22,95%CI(-8.11,-4.34)],and the first defecation(P<0.001,MD=-11.08,95%CI(-15.78,-6.39)]than the control group. Conclusion:EA treatments can promote the recovery of gastrointestinal function after LC.

Objective:To discuss the point-selection and point-grouping patterns and therapeutic application features in acupuncture-moxibustion treatment of acute gouty arthritis(AGA)based on complex networks and to provide references for treating AGA with acupuncture-moxibustion therapy. Methods:Articles related to acupuncture-moxibustion treatment of AGA were searched across the China National Knowledge Infrastructure(CNKI),Chongqing VIP Database(CQVIP),Wanfang Data Knowledge Service Platform(Wanfang),Chinese Biomedical Literature Service System(SinoMed),PubMed,Web of Science(WOS),and Excerpta Medica Database(EMBASE)from their inception till March 31,2023.An acupuncture-moxibustion prescription database was established after the articles were screened according to the inclusion and exclusion criteria.The association rule and complex network analyses of points were conducted using SPSS Modeler 18.1 and Gephi 0.9.7. Results:A total of 145 articles were collected,contributing 382 pieces of acupuncture-moxibustion prescriptions involving 104 points with a total frequency of 1 288.Ashi points contributed the highest frequency.The Spleen Meridian of Foot-Taiyin and the Stomach Meridian of Foot-Yangming were more commonly selected.Filiform-needle acupuncture and bloodletting therapy were more frequently used.The association rule analysis revealed that the highest degree of support belonged to"Ashi point-Zusanli(ST36)"and"Sanyinjiao(SP6)-Zusanli(ST36)",which reflected the rules of point combination of distal and proximal areas and point combination of the coupled meridians.The complex network analysis of the major points discovered a core point prescription mainly consisting of Ahi point,Zusanli(ST36),Sanyinjiao(SP6),Yinlingquan(SP9),and Taichong(LR3).Pattern differentiation and region differentiation were used in selecting adjunct points,stressing the improvements of patterns and joint-related symptoms. Conclusion:Acupuncture-moxibustion treatment of AGA follows the principle of combining major points with adjunct points selected based on pattern or region differentiation;the selection of major points focuses on regulating the deficient Zang-Fu organs,and the selection of adjunct points emphasizes improving patterns and symptoms.The specificity of therapeutic effects is also stressed.

AIM:To investigate the role and possible mechanisms of disulfiram(DSF)in a rat model of heart failure with preserved ejection fraction(HFpEF)induced by high-fat diet(HFD)and nitric oxide blocker Nω-nitro-L-argi-nine methyl ester(L-NAME).METHODS:The HFpEF rat model was constructed using HFD and L-NAME.Sprague-Dawley rats were randomly divided into 3 groups:control group(fed with a normal diet and water),HFpEF group(fed with HFD and drinking water containing 0.5 g/L L-NAME),and DSF+HFpEF group(treated with DSF in addition to HFD and L-NAME).After 5 weeks,cardiac function of the rats was examined using echocardiography and exercise test.Myo-cardial pathological changes were detected using hematoxylin-eosin and wheat germ agglutinin staining,the degree of car-diac fibrosis was assessed using Masson staining,and apoptosis levels were observed using TUNEL staining.Western blot was performed to detect the expression of nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3),cleaved caspase-1,gasdermin D N-terminal fragment(GSDMD-N)in the myocardium,and serum level of N-terminal pro-brain natriuretic peptide(NT-proBNP),and interleukin(IL)-1β and IL-18 in the myocardium were detected by ELISA.RESULTS:Compared with control group,the rats in HFpEF group showed increased body weight,systolic blood pres-sure,diastolic blood pressure,E/E′ ratio,left ventricular anterior wall thickness at diastole and serum NT-proBNP level(P<0.05),and decreased E/A ratio and absolute value of global longitudinal strain(GLS;P<0.05).In contrast,the rats in DSF+HFpEF group showed decreased body weight,E/E′ ratio,diastolic blood pressure and serum NT-proBNP level(P<0.05),and increased E/A ratio and absolute value of GLS(P<0.05),with no significant changes in systolic blood pressure,left ventricular posterior wall thickness at diastole and left ventricular ejection fraction(P>0.05).The rats in HFpEF group had increased myocardial fibrosis area,cardiomyocyte cross-sectional area,and apoptotic rate compared with control group(P<0.05),while these indexes were reduced in DSF+HFpEF group(P<0.05).The results of Western blot and ELISA showed that the levels of NLRP3,cleaved caspase-1,GSDMD-N,IL-1β and IL-18 were increased in the myocardium of rats in HFpEF group compared with control group(P<0.05),but decreased in DSF+HFpEF group com-pared with HFpEF group(P<0.05).CONCLUSION:Disulfiram improves cardiac function and attenuates myocardial remodeling in HFpEF rats.The mechanism may be related to the modulation of NLRP3/caspase-1/GSDMD signaling path-way and the reduction of myocardial inflammatory response.

Objective:To develop the Fecal Microbiota Transplantation Knowledge, Attitude, and Practice Scale for Patients with Inflammatory Bowel Disease (IBD), and test its reliability and validity.Methods:Guided by the theory of knowledge, attitude, and practice, a preliminary draft of the scale was formed through literature review, Delphi expert consultation, and pre-survey. From May to August 2022, convenience sampling was used to select 200 IBD patients who visited the Gastroenterology Clinic of three ClassⅢ Grade A comprehensive hospitals in Jiangsu Province as the research subject for a questionnaire survey. The critical ratio method, correlation analysis method, internal consistency method, commonality and factor loadings were used for item analysis of the scale. Exploratory factor analysis, content validity index, and internal consistency reliability were applied to test the reliability and validity of the scale.Results:A total of 200 questionnaires were distributed, and 181 valid questionnaires were collected, with an effective response rate of 90.50% (181/200). The Fecal Microbiota Transplantation Knowledge, Attitude, and Practice Scale for Patients with IBD included three dimensions of knowledge, attitude and practice, with a total of 21 items. The content validity index at the scale level was 0.917, and the content validity index at the item level ranged from 0.833 to 1.000. Exploratory factor analysis extracted three common factors, with a cumulative variance contribution rate of 74.197%. The Cronbach's α coefficient of the total scale was 0.951, and the coefficients of each dimension were 0.914 to 0.942. The test-retest reliability coefficient of the total scale was 0.918, and the test-retest reliability coefficients of each dimension ranged from 0.737 to 0.833.Conclusions:The Fecal Microbiota Transplantation Knowledge, Attitude, and Practice Scale for Patients with IBD has good reliability and validity, which can help medical and nursing staff evaluate patients' understanding and acceptance of microbial transplantation, so as to provide a basis for personalized communication in shared decision making between doctors and patients.