Objective To construct CAR-T cells harboring distinct intracellular signaling domains, thereby addressing the core limitations of conventional CAR-T cells, including premature exhaustion, insufficient antitumor efficacy, and uncontrolled cytokine release syndrome caused by excessive intracellular activation signals. Methods The intracellular signaling domain CD3ζ of the conventional CAR construct was replaced with the CD3ε signaling domain containing only one immunoreceptor tyrosine-based activation motif （ITAM）, to generate a novel BCMA-E-CAR-T cell product. Flow cytometry was employed to assess the cytotoxicity against target cells, activation status, exhaustion profile, central memory phenotype differentiation, proliferative capacity, and cytokine secretion levels of the engineered CAR-T cells. Results BCMA-E-CAR-T cells exhibited significantly enhanced cytotoxicity against target cells, accompanied by reduced activation levels, alleviated exhaustion, diminished cytokine release, elevated differentiation toward a central memory phenotype, and stronger proliferative potential. Conclusion This study successfully constructed novel BCMA-E-CAR-T cells harboring CD3ε as the intracellular activation signaling domain, which exhibited significantly enhanced antitumor activity and improved therapeutic safety in tumor models.

Objective To investigate the role and mechanism of spermidine/spermine N1-acetyltransferase 1 （Sat1） in cisplatin （DDP）-induced injury of mouse cochlea-derived immortalized auditory epithelial cells （HEI-OC1）. Methods HEI-OC1 cells were divided into four groups： siNC, siSat1, siNC+DDP, and siSat1+DDP. Cell proliferation was measured by CCK-8 assay; intracellular reactive oxygen species （ROS） levels were detected using DCFH-DA fluorescent probe; mRNA expression levels of Sat1, catalase （CAT）, and glutathione peroxidase 1 （GPx1） were assessed by qPCR; protein expression of Sat1 and apoptosis-related proteins, including B-cell lymphoma-2 （Bcl-2）, Bcl2-associated X protein （Bax）, cysteinyl aspartate-specific proteinase-3 （Caspase-3）, and cleaved Caspase-3, were detected by Western blotting, and the ratios of cleaved Caspase-3/Caspase-3 and Bcl-2/Bax were calculated. Malondialdehyde （MDA） content and superoxide dismutase （SOD） activity were measured by ELISA to evaluate oxidative stress. Results Compared with the siNC group, the siSat1 group showed downregulated Sat1 mRNA and protein expression, decreased basal ROS and MDA levels, increased SOD activity and CAT and GPx1 mRNA expression, and reduced Bax and Cleaved Caspase-3 protein expression （all P < 0.05）. After DDP treatment, the siNC+DDP group exhibited decreased cell viability, elevated ROS and MDA levels, reduced SOD activity and CAT and GPx1 mRNA expression, and increased Bax and cleaved Caspase-3 protein expression; all these alterations were significantly reversed in the siSat1+DDP group （all P < 0.05）. Conclusion Sat1 plays a critical role in DDP-induced HEI-OC1 cell apoptosis by mediating oxidative stress.

Objective To explore the effect of long non-coding RNA Vof16 （lncRNA Vof16） on axonal regeneration after spinal cord injury （SCI） via the heterogeneous nuclear ribonucleoprotein U （HNRNPU）/complexin-1 （CPLX1） axis. Methods In vivo, adult SD rats were randomly allocated to 6 groups： Sham group, SCI group, SCI+sh-NC group, SCI+Vof16-KD group, SCI+oe-NC group and SCI+Vof16-OE group. Motor and sensory functions were evaluated with BBB scores and mechanical withdrawal thresholds. Differentially expressed lncRNAs were screened by RNA-seq and validated by qRT-PCR; spinal-cord levels of Vof16, HNRNPU and CPLX1 were quantified by qRT-PCR. In vitro, primary spinal cord neurons were isolated for oxygen glucose deprivation （OGD）, Vof16 over-expression （Vof16-OE）, HNRNPU over-expression （HNRNPU-OE） or knock-down （HNRNPU-KD）, and Vof16-OE+ HNRNPU-OE and Vof16-OE+ HNRNPU-KD groups were constructed.Cell proliferation, migration and microtubule stability （Ace-tubulin/Tyr-tubulin） were assessed with CCK-8, Transwell assay and immunofluorescence, respectively. The Vof16-HNRNPU-CPLX1 interaction was verified by RNA pull-down, RIP and CLIP assays. Results RNAseq results showed that lncRNA Vof16 was highly expressed in spinal cord tissue after SCI in rats. Over-expressing Vof16 in OGD neurons increased proliferation and migration, raised ace-tubulin and reduced tyr-tubulin （P < 0.05）. Mechanistically, Vof16 directly bound HNRNPU and prevented its degradation; HNRNPU in turn bound CPLX1 mRNA and enhanced its stability. HNRNPU knock-down abolished the pro-regenerative effects of Vof16. In vivo, BBB scores at 21st day post-injury were higher and mechanical withdrawal thresholds were lower in the SCI+Vof16-OE group than in the SCI+oe-NC group （P < 0.05）, accompanied by elevated spinal-cord expression of HNRNPU and CPLX1. Conclusion lncRNA Vof16 recruits HNRNPU to stabilize CPLX1 mRNA, thereby promoting axonal regeneration after SCI.

Objective The intestinal microbiota is related to the primary site of colorectal cancer （CRC）. To explore the differences in the intestinal microbiome among different primary sites of colorectal cancer, this study conducted a meta-analysis using comprehensive genomic data. Methods Genomic data were collected from three studies and grouped according to three sites： right-sided colon cancer （RCC）, left-sided colon cancer （LCC）, and rectal cancer （RC）. Firstly, confounding factors were identified through two-factor variance analysis and comparison of microbial diversity. Subsequently, a microbial co-occurrence network was constructed. A random forest algorithm was used to build a prediction model for the location of the primary tumor. Finally, tumor location and confounding factors were incorporated into the Maaslin2 （microbiome multivariable associations with linear models） model and linear discriminant analysis （LDA） was used to identify differentially expressed species. Results The sources of research and BMI are the main confounding factors. There is no significant difference in the α/β diversity of intestinal microbiota among different tumor sites. The Actinobacteria phylum, Firmicutes phylum, and Proteobacteria phylum all play key linking roles in the three microbial networks, and the Bacteroidetes phylum is more significant in the microbial network of RCC. There are both common key bacterial groups and their own unique key bacterial groups in the three networks. The random forest classification model performs well in predicting RC, but performs poorly in distinguishing RCC from LCC. By comparing CC with RC, and comparing RCC with LCC, some of the differential bacterial groups identified by Maaslin2 and LDA have been reported to be related to molecular and immune characteristics of CRC location. Conclusion This study elucidated the relationship between gut microbiome and CRC location and confirmed that RCC, LCC, and RC had different enrich patterns of microbiota.

Objective To investigate the role of sialic acid-bound immunoglobulin-like lectin 15 （Siglec15） in the occurrence and development of hepatocellular carcinoma （HCC） and its underlying mechanism. Methods Online databases were utilized to analyze the expression pattern of Siglec15 in HCC, followed by validation in HCC cell lines. HCC cell models with Siglec15 overexpression or knockdown were established to evaluate the regulatory effects of Siglec15 on the proliferation and migration of HCC cells. A subcutaneous tumor xenograft model in nude mice was constructed to observe the impact of Siglec15 overexpression on tumor growth in vivo. Transcriptome sequencing combined with KEGG pathway enrichment analysis was performed, and the related signaling pathways were further verified by qRT-PCR and Western blotting. Moreover, the expression levels of key molecules in the PI3K/AKT signaling pathway （p-PI3K, p-AKT） as well as proteins associated with cell cycle regulation and apoptosis （P53, Caspase3, Bax, Bcl-2） were detected. Results Compared with that in the adjacent normal liver tissues, Siglec15 was significantly upregulated both in the HCC tissues and cell lines （P<0.05）. Knockdown of Siglec15 markedly suppressed the proliferation and migration capacities of the HCC cell line SMMC-7721 （P<0.05）. The nude mouse xenograft experiment demonstrated that Siglec15 overexpression promoted tumor growth in vivo. Transcriptome sequencing identified 4 765 differentially expressed genes, among which the expression alterations of core moleculars involved in the PI3K-AKT pathway （PI3K, AKT, Bcl-2, CCNA2, P53, etc.） were highly consistent with the observed cellular phenotypes. Validation experiments confirmed that Siglec15 overexpression upregulated the mRNA expression of PI3K, AKT, Bcl-2 and CCNA2 while downregulating that of P53, Caspase3 and Bax. Meanwhile, Siglec15 overexpression significantly increased the phosphorylation levels of PI3K and AKT proteins, with concomitant modulation of the expression of downstream apoptosis and cell cycle-related proteins. Conversely, the Siglec15 knockdown group exhibited the opposite trends. Conclusion Siglec15 promotes the proliferation and migration of HCC cells by activating the PI3K/AKT signaling pathway, primarily via post-translational phosphorylation modification, which in turn regulates the expression of downstream apoptosis- and cell cycle-associated proteins. Therefore, Siglec15 may serve as a potential therapeutic target for HCC.

Objective To investigate the regulatory effect and potential mechanism of lentiviral vector-mediated microRNA-195 （miR-195） downregulation on osteogenic and angiogenic differentiation of bone marrow mesenchymal stem cells （BMMSCs）. Methods Human BMMSCs were divided into three groups： blank control group, inhibitor-negative control （NC） group （transfected with lentiviral NC sequence）, and miR-195-inhibitor group （transfected with lentiviral miR-195 inhibitor sequence）. The expression and localization of miR-195 were detected by RNA fluorescence in situ hybridization. Cell proliferation was measured by CCK-8 assay. Osteogenic differentiation was evaluated by alkaline phosphatase （ALP） staining, alizarin red staining, and Runt-related transcription factor 2 （Runx2） immunofluorescence. Tube formation assay was performed for angiogenesis assessment. The mRNA levels of vascular endothelial growth factor （VEGF）, ALP, Runx2, VEGF receptor 2 （VEGFR2）, angiopoietin-1 （Ang-1）, and Wnt/β-catenin pathway-related molecules （Wnt3a, Wnt10b, β-catenin, Osterix） were determined by qRT-PCR. The protein expression of the above pathway and osteogenesis-related molecules was detected by Western blotting. Results Compared with the control groups, the miR-195-inhibitor group showed significantly decreased miR-195 expression （all P<0.05）, and markedly increased cell proliferation, ALP activity （4.15±0.52 vs 1.00±0.15）, alizarin red positive rate （64.67%±7.78% vs 31.86%±2.24%）, Runx2 fluorescence intensity （852.38±122.70 vs 121.31±12.16 RFU）, and tubular structure length （218.60±24.75 vs 49.80±5.25 μm/field）. The mRNA levels of VEGF, ALP, Runx2, VEGFR2, Ang-1, Wnt3a, Wnt10b, β-catenin, and Osterix, as well as the protein levels of Wnt3a, Wnt10b, β-catenin, and Osterix were all significantly upregulated （all P<0.05）. There were no significant differences in the above indices between the inhibitor-NC group and the blank control group （P>0.05）. Conclusion Lentiviral-mediated miR-195 downregulation synergistically promotes osteogenic and angiogenic differentiation of BMMSCs via activating the Wnt/β-catenin signaling pathway.

Objective To investigate the molecular mechanism through which resveratrol （RES） modulates the progression of diabetic foot by targeting SRC proto-oncogene, non-receptor tyrosine kinase （SRC）/fibroblast growth factor receptor 1 （FGFR1） axis. Methods Human dermal fibroblasts （HDF） were allocated into NG （normal glucose）, HG （high glucose）, and HG+RES groups. Cell proliferative viability was measured by CCK-8 assay, migration by Transwell assay, and apoptosis by TUNEL staining. Bioinformatic analyses were performed to predict potential targets of RES in diabetic foot and to explore the associated signaling pathways. HDF cells were further divided into NG, HG, HG+si-SRC, HG+oe-SRC, and HG+oe-SRC+si-FGFR1 groups to determine how alterations in SRC and FGFR1 expression affect proliferative viability, migratory capacity, and apoptosis. A diabetic foot animal model was established and treated with RES; α-lipoic acid served as the positive control to evaluate the therapeutic potential of RES. Results Bioinformatic screening identified SRC as one of the intersecting targets of RES and diabetic foot, and as a key molecule in endocrine resistance pathway. Compared with the NG group, HG-treated HDF cells exhibited up-regulated SRC expression, reduced proliferative viability and migratory capacity, and increased apoptosis; these effects were reversed by RES intervention （P<0.05）. Relative to the HG group, SRC knockdown enhanced cell viability and migration while decreasing apoptosis, whereas SRC overexpression further suppressed proliferative viability and migration and promoted apoptosis; simultaneous FGFR1 knockdown partially reversed these latter effects （P<0.05）. Animal experiments confirmed that both RES and α-lipoic acid exert therapeutic efficacy in the diabetic foot model. Conclusion Resveratrol promotes the improvement of diabetic foot by inhibiting the SRC/FGFR1 axis.

Objective To investigate the clinical significance of the expression of special AT-rich sequence-binding protein 2 （SATB2）, cytokeratin 20 （CK20）, and mucin 2 （MUC2） in colorectal cancer （CRC） tissues and their value in prognostic evaluation. Methods A total of 330 CRC patients who underwent radical resection at Hubei Provincial Hospital of Integrated Traditional Chinese and Western Medicine from March 2020 to March 2022 were retrospectively analyzed. After excluding 15 patients lost to follow-up, 315 patients were included in the survival analysis. CRC tissues and paired adjacent normal tissues were collected. Immunohistochemistry was used to detect the expression of the three proteins, and patients were grouped accordingly. The relationships between protein expression and clinicopathological features were analyzed. Kaplan-Meier survival analysis and Cox regression models were used to evaluate prognostic factors. Results The positive expression rates of SATB2 and MUC2 in CRC tissues were lower than those in adjacent tissues, while CK20 expression was higher （P<0.05）. Patients with tumor diameter <4 cm, TNM stage Ⅰ~Ⅱ, and no lymph node metastasis showed higher positive rates of SATB2 and MUC2 and lower CK20 positivity （all P<0.05）. Univariate and multivariate Cox regression analyses indicated that tumor diameter ≥4 cm, TNM stage III-IV, lymph node metastasis, and CK20 positivity were risk factors for poor prognosis, whereas SATB2 and MUC2 positivity was protective factors （all P<0.05）. Survival analysis showed that patients positive for SATB2 and MUC2 had lower 3-year mortality, while those positive for CK20 had higher mortality （P<0.05）. Conclusion The down-regulation of SATB2 and MUC2 expression in CRC tissue is closely related to tumor progression, and their positive expression suggests a better prognosis. The positive expression of CK20 may be related to poor prognosis. The expression status of SATB2, CK20 and MUC2 can be used as potential supplementary markers to judge the prognosis risk of CRC patients.

Objective To investigate the relationship between serum interleukin-17 （IL-17）, cathelicidin antimicrobial peptide LL-37 levels and carotid atherosclerosis in patients with moderate-to-severe plaque psoriasis. Methods A total of 168 patients with moderate-to-severe plaque psoriasis admitted between June 2023 and December 2024 were selected and serum IL-17 and LL-37 levels were measured.According to cartid intima-media thickness（IMT）, patients were divided into atherosclerosis group （IMT ≥1.5 mm, n=56） and non-atherosclerosis group （IMT <1.5 mm, n=112）. Clinical data and serum IL-17 and LL-37 levels were compared between the two groups. We used Pearson correlation analysis to evaluate the association between serum IL-17, LL-37 levels and IMT. Additionally, logistic regression analysis was applied to determine the factors affecting carotid atherosclerosis. Additionally, receiver operating characteristic （ROC） curve analysis was conducted to evaluate the diagnostic efficacy of serum biomarkers for carotid atherosclerosis. Results The Psoriasis Area and Severity Index score and levels of low-density lipoprotein cholesterol （LDLC）, C-reactive protein, uric acid, IL-17 and LL-37 in serum of the atherosclerosis group were higher than those of the non-atherosclerosis group （P<0.05）. Serum IL-17 and LL-37 showed significant positive correlations with carotid IMT （P<0.05）. Elevated IL-17 （OR=1.255）, elevated LL-37 （OR=1.014）, and elevated LDLC （OR=13.278） were risk factors for carotid atherosclerosis （P<0.05）. The areas under the curve （AUC） of IL-17, LL-37, LDLC for diagnosing carotid atherosclerosis were 0.861, 0.848, 0.702, respectively. The AUCs for IL-17 and LL-37 in diagnosing carotid atherosclerosis were higher than those for LDLC （P<0.05）, and there was no statistically significant difference in the AUCs between IL-17 and LL-37 （P>0.05）. Conclusion Elevated serum IL-17 and LL-37 levels are associated with carotid atherosclerosis in patients with moderate-to-severe plaque psoriasis. The diagnostic efficacy of IL-17 and LL-37 as diagnostic markers for carotid atherosclerosis is superior to that of LDLC.

Objective This study aimed to elucidate the correlation between serum levels of circ_UTRN and circ_ZFP644 and disease severity in patients with acute pancreatitis （AP）. Methods A cohort of 188 AP patients admitted between October 2020 and August 2025 was enrolled as the AP group, while 90 healthy individuals undergoing routine physical examinations served as the control group. Serum expression levels of circ_UTRN and circ_ZFP644 were quantified using quantitative real-time polymerase chain reaction. Based on clinical severity criteria, AP patients were stratified into mild, moderately severe, and severe subgroups for comparative analysis, and into good prognosis group and poor prognosis group according to the prognosis. The correlations between the expression levels of serum circ_UTRN and circ_ZFP644 in AP patients and the bedside index for severity in acute pancreatitis （BISAP） score and the acute physiology and chronic health evaluation Ⅱ （APACHE Ⅱ） score, the independent factors influencing the prognosis of AP patients, the predictive capacity of serum circ_UTRN and circ_ZFP644 expression levels for prognosis were analyzed. Results The expression levels of serum circ_UTRN and circ_ZFP644 in the AP group were lower than those in the control group, with statistically significant differences （P<0.05）. The expression levels of circ_UTRN and circ_ZFP644 in the serum of the AP group were as follows： mild group > moderately severe group > severe group（P<0.05）. The expression levels of serum circ_UTRN and circ_ZFP644 in AP patients were negatively correlated with the BISAP score and the APACHE II score （P<0.05）. The expression levels of serum circ_UTRN and circ_ZFP644 in AP patients with poor prognosis were lower than those in patients with good prognosis, while the BISAP score, APACHE II score, C-reactive protein level, and procalcitonin level were higher than those in patients with good prognosis, with statistically significant differences （P<0.05）. Serum circ_UTRN and circ_ZFP644 were protective factors for a poor prognosis in AP, while the BISAP score and APACHE II score were risk factors for a poor prognosis in AP （P<0.05）. The area under the curve for the combined prediction of AP prognosis by serum circ_UTRN and circ_ZFP644 was higher than that of either indicator alone（P<0.05）. Conclusion The low expression levels of serum circ_UTRN and circ_ZFP644 are significantly associated with disease progression and poor prognosis in AP patients, the combined detection has predictive value for the prognosis of AP.

Objective To screen extracellular matrix （ECM） proteins associated with neointimal hyperplasia （NH） in arteriovenous graft （AVG） fistula using bioinformatics methods, explore the underlying molecular mechanisms, and provide novel molecular targets and drug candidates for clinical treatment. Methods The dataset GSE97377 was obtained from the GEO database. Transcriptomic data on day 14 after AVG modeling served as the training set, and data on day 5 served as the validation set. Differentially expressed genes （DEGs） were identified using the limma package. ECM proteins were extracted from the Uniprot and HPA databases, and intersection with DEGs was performed to obtain ECM protein-related DEGs （EP-DEGs）. GO and KEGG pathway analyses were conducted on EP-DEGs. A protein-protein interaction network was constructed to screen for hub genes. Functional analysis of EP-DEGs was performed using the GeneMANIA database. Drug prediction and molecular docking were carried out using the DSigDB and CB-DOCK2 databases, respectively. Results A total of 79 EP-DEGs associated with NH development after AVG fistula were identified, among which IL1A, CXCL12, CD163, and SPP1 were identified as key hub genes. GO and KEGG analyses indicated that these genes were primarily involved in inflammatory response and ECM remodeling. The expression levels of CXCL12, CD163, and SPP1 were consistent in the venous vasculature on both day 5 and day 14 after AVG modeling. GeneMANIA analysis suggested that CXCL12 is a key regulator of inflammatory factors. Drug prediction analysis revealed that simvastatin, felodipine, kaempferol, and vorinostat may exert therapeutic effects by targeting the hub genes. Molecular docking analysis demonstrated strong interactions between the target proteins and candidate drugs. Conclusion This study systematically identified ECM proteins associated with NH after AVG fistula and predicted multiple potential molecular targets and candidate drugs, providing a theoretical basis for understanding the molecular mechanisms of NH and developing novel therapeutic strategies.

Post-translational modifications serve as key regulatory mechanisms in functional proteomics, extensively involved in modulating protein activity, subcellular localization, and interactions. Among these, lysine crotonylation is a novel form of modification discovered in recent years, occurring on both histones and non-histone proteins. It participates in critical biological processes such as gene transcription, cell cycle progression, and DNA damage response. This review systematically summarizes the regulatory roles and molecular mechanisms of lysine crotonylation in cardiovascular diseases, respiratory diseases, neurological disorders, as well as kidney diseases, diabetes, autoimmune diseases, and reproductive endocrine-related conditions, aiming to provide a new theoretical foundation and research perspectives for targeted therapeutic strategies of related diseases.

Monogenic obesity is obesity caused by a mutation in a single gene, accounting for about 2% to 10% of obesity in children and adults. Since the identification of the leptin gene and its receptor, over 85 forms of monogenic obesity have been discovered, with the vast majority of pathogenic genes concentrated in the hypothalamic leptin-melanocortin signaling pathway, which is a key hub in the central regulation of appetite and energy balance. Mutations in the MC4R gene are the most common cause of monogenic obesity, accounting for more than a quarter of all cases. In recent years, with the advancement of whole-exome sequencing and large-scale cohort studies, new genes such as TUB, ADCY3, and ALMS1 have been identified, greatly expanding the genetic spectrum of monogenic obesity. In terms of clinical treatment, precision drugs based on genetic mechanisms have made breakthrough progress： the MC4R agonist setmelanotide has been approved for specific genetic subtypes such as POMC, PCSK1, LEPR deficiencies, and Bardet-Biedl syndrome, while the oral MC4R modulator CGX-926 has entered Phase I clinical trials. This article provides a systematic overview of the pathogenic gene spectrum, molecular mechanisms, genetic testing strategies, and targeted treatment progress in monogenic obesity, aiming to provide a theoretical basis for precise clinical diagnosis and treatment.

Porto-sinusoidal vascular disease （PSVD） is a recently redefined vascular liver disorder characterized by structural injury of the porto-sinusoidal microcirculation and consequent complications related to portal hypertension. With advances in histopathology, transcriptomics, genetic studies, and medical imaging technologies, the diagnostic approach to PSVD has evolved from a primarily exclusion-based clinical diagnosis toward a precision-oriented strategy integrating pathological assessment, molecular profiling, and non-invasive evaluation. This review summarizes recent progress in the pathophysiological and molecular mechanisms, histopathological features, clinical manifestations, molecular and non-invasive diagnostic clues, and therapeutic strategies of PSVD. Furthermore, we discuss future perspectives regarding the development of stratified diagnostic models and mechanism-directed therapeutic approaches for this disease.

Gene editing technology can precisely modify specific DNA segments in the genome of an organism, opening up new ways for disease treatment, gene function research, gene testing and drug development. Conventional CRISPR-Cas9 gene editing relies on DNA double-strand break（DSB）, whose repair processes are prone to inducing indel mutations, exhibit cytotoxicity, and demonstrate low efficiency in homology-directed repair, significantly limiting its applications in precision medicine. To overcome this limitation, a series of novel CRISPR-derived technologies independent of DSB have emerged. This review systematically summarizes the principles, development, and applications of three representative technologies： base editors, prime editors, and transposon-based editors, with a focus on analyzing how their mechanistic innovations circumvent DSB. It further explores their respective advantages and inherent limitations in terms of editing efficiency, precision, target scope, and delivery challenges. Finally, the review offers perspectives on future technological directions, highlighting that editor miniaturization, efficiency enhancement, and delivery tool innovation are key to advancing clinical translation in this field.

Objective To address the growing demand for cultivating high-level innovative talents under the "New Medicine" initiative, this study aimed to resolve existing challenges in the traditional teaching of Molecular Biology, such as outdated course content, limited diversity in teaching staff, and the disconnection between theory and practice, thereby exploring an effective pathway to shift the teaching focus from knowledge transmission to fostering innovation capability and scientific literacy. Methods A teaching reform centered on a "Module + 3X" framework was implemented in the Molecular Biology course for master's students at Air Force Medical University. With the flow of genetic information as the main thread, four modular teaching units were constructed： "The Central Dogma： Everlasting and Renewed," "Biotechnology： Rapidly Evolving," "Molecular Research： In-depth Exploration," and "Clinical Translation： Mutual Enhancement." This design enabled a stepwise progression from classical theories to cutting-edge advances. The "3X" component integrated thematic lectures, master classes, and case sharing by young scholars to promote multidisciplinary integration and strengthen the link between theory and practice. The effectiveness of the reform was systematically evaluated using customized questionnaires and formative assessments. Results Following the implementation of the reform, significant improvements were observed in course selection rate, course satisfaction, students' knowledge mastery, and their interest in scientific research. Formative assessment results indicated that the "Module + 3X" system effectively enhanced students' understanding of both classical theories and advances in molecular biology, while also strengthening their interdisciplinary thinking and practical skills. Conclusion The "Module + 3X" teaching system successfully facilitated the transformation of Molecular Biology instruction from knowledge delivery to the cultivation of innovation ability and scientific literacy, markedly improving teaching outcomes and the quality of postgraduate education.