Current Biotechnology

Biological Breeding Science and Technology Innovation for Seed Industry Development and Prospects
Zhenhua XU, Shiwei GAO, Dawei GAO, Yanming YU, Haiying LIU, Hongtao WU, Shuli ZHANG, Zhongyi SUN, Xin WANG, Ping YAN
2025, 15(4): 557-564. DOI: 10.19586/j.2095-2341.2025.0036
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In recent years， China has achieved significant progress in the application of biological breeding technologies. However， there remains considerable scope for breakthroughs in basic theoretical research， core technology development， and market cultivation of major varieties. Biological breeding is a core driver of innovation in the modern bio-breeding industry， which innovates breeding techniques， methods， and products， thereby enhancing the yield， quality， and adaptability of crops. These improvements aim to meet the increasing global demand for food and address environmental challenges. Developed countries and international seed industry giants have integrated biotechnology with big data， artificial intelligence， and other cutting-edge technologies to propel the seed industry into the "Seed Industry 4.0" era， and the breeding mode is shifting from traditional "hybrid selection" to "intelligent selection and breeding". Based on this， the article reviewed the development history of biological breeding technology， the current status of the domestic and international biological breeding industry， summarized the innovation and application of key technologies in biological breeding， discussed the problems and challenges facing the industrialization of biological breeding， and looked forward to future development trends， in order to provide reference for enhancing China's biological breeding innovation capabilities and seed industry development.

Current Status of Research Application and Safety Regulation of Plant Bioreactors
Jiangtao YANG, Yaohui HUANG, Zhixing WANG, Xujing WANG, Yue JIAO
2025, 15(4): 565-572. DOI: 10.19586/j.2095-2341.2024.0202
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Plant-derived bioreactors which produce pharmaceutical and industrial bioproducts utilizing transgenic technology represent one of the fastest-growing categories of biotechnological products in research and application. Plants are increasingly employed as biofactories to produce high-quality biopharmaceuticals， including antibodies， vaccines， therapeutic proteins， hormones， and cytokines， along with bioproducts for cosmetics， food， and the chemical industry. These products serve as medicines and biomaterials in pharmaceutical and industrial sectors. This article explored the significant potential of transgenic plants as production platforms， reviewed recent research advances in plant-derived bioproducts， and highlighted their multifunctional applications across diverse fields such as medicine， industry， and agriculture. It also examined the safety evaluation frameworks and regulatory policies governing such plants in regions including the United States and the European Union. This review aimed to provide insights for enhancing safety assessment and regulation of medicinal and industrial transgenic plants in China.

Research Progress on the Enzymatic Properties of Common Enzyme Preparations in Soybean Meal Enzyme Hydrolysis Processes
Chuang LIU, Jianshuang MA, Yingguo BAI, Huoqing HUANG, Tao TU, Huimin YU, Ning LIU, Yuan WANG
2025, 15(4): 573-586. DOI: 10.19586/j.2095-2341.2025.0055
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Soybean meal serves as an important plant protein source widely utilized in feed and food industries. However， the presence of anti-nutritional factors （such as antigenic protein and non-starch polysaccharides） limits its nutritional value. Enzymatic hydrolysis has emerged as a novel in vitro pretreatment method to enhance the nutritional quality of soybean meal， with commonly used enzymes including proteases， cellulases， and galactosidases. The review systematically summarzied the types of anti nutritional factor in soybean meal， different enzyme preparations and their mechanisms of action， and the research process on the application of enzyme preparations， aiming to provide theoretical foundations and technical references for enzyme selection in soybean meal enzymatic hydrolysis processes.

Research Advances on Chemical Constituents, Pharmacological Effects, Extraction Analytical and Processing Techniques of Star Anise
Qin WEI, Danna HUANG, Lu CHEN, Cong WANG
2025, 15(4): 587-596. DOI: 10.19586/j.2095-2341.2025.0025
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Star anise （Illicium verum Hook. f.） belongs to the Magnoliaceae family， a medicinal-food homologous plant endemic to China， has current research primarily focused on the development and utilization of its chemical components and pharmacological effects. However， challenges remain in the systematic identification of its chemical composition， insufficient quantitative data on pharmacological effects， and the absence of standardized protocols for processing and extraction technologies. The review systematically sorted out the main constituents， including volatile oils， phenolic acids， flavonoids， and sesquiterpene lactones， analysed the pharmacological activities such as antibacterial， analgesic， anti-inflammatory， and antioxidant effects and summarized the main points of extraction techiniques， analysis and fresh material processing technology. It provides strong support for improving the research methods and technological standards of star anise， improving the efficiency of resource utilization and industrial application.

Application Progress of Biosensors Based on Field-effect Transistors in Nucleic Acid Detection
Yue SHI, Yao HAN, Hao LI, Yansong SUN
2025, 15(4): 597-605. DOI: 10.19586/j.2095-2341.2024.0204
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The field-effect transistor （FET） biosensor is a type of sensor based on the principle of detecting biomolecules or biomarkers. Due to the signal amplification characteristics of its electric field effect， it enables the detection of biomolecular interactions at extremely low concentrations， further enabling biomolecular recognition. In the field of nucleic acid detection， this approach offers the unique advantage of eliminating the need for nucleic acid amplification， resulting in shorter detection times. Additionally， FET biosensors also excel in nucleic acid detection performance， portability， and cost-effectiveness. Overall， FET biosensors can achieve ultra-rapid and ultra-sensitive targeted identification of viral genes. Combined with microfluidic technology， nanomaterials， or flexible electronic materials， they hold significant promise for application in food safety， clinical diagnosis， genotyping， biosafety， and other fields. To facilitate the better application of FET sensors in biological detection， this review summarized the current applications of FET biosensors in nucleic acid detection and provided a classification and discussion based on the biological recognition layer， aiming to offer insights for future research in this area.

Research Progress of Immunomagnetic Separation Technology in Detection of Circulating Tumor Cell
Lulu ZHAO, Tian HONG, Yiran HAO, Erning CHEN, Jingwen LI, Meihong DU
2025, 15(4): 606-614. DOI: 10.19586/j.2095-2341.2025.0030
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Tumor invasion and metastasis are the primary causes of death in cancer patients. Circulating tumor cell （CTC）， as the seed cells of tumor metastasis， carries complete cellular biological information and hold significant importance in early diagnosis， prognosis assessment， and personalized treatment monitoring of tumors. However， due to the extremely low number of CTC in the blood， developing efficient enrichment and separation methods are crucial for the precise analysis of CTC. Immunomagnetic separation technology， with its advantages of high specificity and efficient enrichment， can effectively and specifically isolate and enrich target substances， providing a powerful tool for biomedical research and clinical diagnosis. As the cross-disciplinary integration and development of materials science， biotechnology， and electronic engineering technology， the application value of immunomagnetic separation technology in cutting-edge analytical areas has been enhanced， expecting to achieve significant breakthroughs in the field of CTC detection. This article reviewed the design of CTC immunomagnetic material， the CTC immunomagnetic separation and capture platform， and the combination of immunomagnetic separation technology with microfluidics， discussed the challenges and optimization strategies of immunomagnetic separation technology in CTC detection， in order to provide a reference for promoting the deeper application of CTC detection technology.

Research Progress on the Cytotoxicity and Immune Effects of Carbon-based Nanomaterials
Yimiao ZHANG, Yuqin BIAN, Xinbo LIU, Jiahe PANG, Tongxuan SUN, Qiazheng DU, Wenhao XU, Tianze YIN, Hongshu SUI
2025, 15(4): 615-621. DOI: 10.19586/j.2095-2341.2025.0024
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Carbon-based nanomaterials have demonstrated extensive application prospects in fields including biomedicine， energy storage， and electronic devices， owing to their unique physicochemical properties. In recent years， significant advancements have been achieved in toxicological studies of these materials， but the mechanisms through which structural variations influence their cytotoxicity and immune effects remain not yet fully elucidated. This review systematically synthesized existing literature， meticulously comparing the performance of diverse carbon nanomaterials in both in vitro cellular assays and in vivo animal models， sorted out the structure-toxicity-immune effect relationships of carbon nanomaterials， and summarized the current research landscapes， development trends and research needs of toxicology research on carbon nanomaterials， in order to provid theoretical basis for their safety design.

Autophagy and Apoptosis: Interactions and Their Role in Disease
Lina ZHU, Zhiling SONG
2025, 15(4): 622-626. DOI: 10.19586/j.2095-2341.2025.0028
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Autophagy and apoptosis are the key physiological processes to maintain cell homeostasis， and their interaction has important pathological significance in tumors， neurodegenerative diseases and cardiovascular diseases. The molecular mechanism of the two dynamic regulatory networks has not been fully analyzed in the existing studies， and the strategies targeting the regulation of autophagy and apoptosis balance in disease treatment are still lacking in accuracy. Studies have shown that through a bidirectional regulatory mechanism and targeting key hub proteins can synergically inhibit apoptosis and activate autophagy. The review summarized the interaction mechanism between artophagy and apoptosis， as well as the research progress in tumors， neurodegenerative diseases， and heart disease， in order to provide a theoretical framework for analyzing cell death regulatory networks and lay an important foundation for developing precise therapeutic strategies based on the balance regulation of autophagy and apoptosis.

Research Progress on Mitochondria Maintaining Skeletal Muscle Homeostasis
Peixian GAO, Xueqi HAN, Jinyi ZHAO, Pengxiang ZHAO, Baihui ZHENG, Mengyu LIU
2025, 15(4): 627-635. DOI: 10.19586/j.2095-2341.2025.0007
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Skeletal muscle homeostasis is essential for muscle health， and its dysregulation can lead to skeletal muscle atrophy and reduce quality of life. It has become a recognized global health problem. Studies have shown that mitochondria plays an important role in maintaining skeletal muscle homeostasis and health， and mitochondrial dysfunction can lead to skeletal muscle atrophy. However， its molecular mechanism is complex and has not been fully elucidated， which hinders the development of therapeutic drugs for skeletal muscle atrophy. This article reviewed the molecular mechanisms of mitochondria maintaining muscle homeostasis， expounded in detail the key roles of mitochondrial biogenesis， mitochondrial dynamics， mitochondrial autophagy and other processes in skeletal muscle health， discussed the impact of mitochondrial dysfunction on skeletal muscle structure and function， and further summarized the methods of targeted regulation of mitochondrial function to treat skeletal muscle atrophy. It is helpful to understand the role of mitochondria in skeletal muscle as a whole， and provide theoretical basis and inspiration for the subsequent development of mitochondria-targeted drugs for the treatment of skeletal muscle atrophy.

Research Progress of Cancer-associated Fibroblasts in Breast Cancer
Xiaoyi ZHAI, Haiyue ZHANG, Wenjia GUO, Xiaogang DONG
2025, 15(4): 636-644. DOI: 10.19586/j.2095-2341.2024.0200
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Cancer-associated fibroblasts （CAFs） are an integral component of the tumor microenvironment. Recent studies have established a close association between CAFs and the development， malignant progression， and poor prognosis of breast cancer. However， a comprehensive understanding of the relationship between CAFs and breast cancer cell growth， invasion， metastasis， and prognosis remains incomplete. In breast cancer， CAFs are regulated by tumor cells. Conversely， CAFs significantly influence tumor behavior by secreting cytokines and exosomes， which modulate the immune microenvironment， remodel the extracellular matrix， and suppress immune cell function. These actions collectively promote tumor initiation， migration， invasion， and drug resistance. This review summarized the role of CAFs in breast cancer cell growth， invasion， and metastasis， as well as their association with patient prognosis. This synthesis aimed to deepen the understanding of breast cancer pathogenesis and provide a theoretical basis for its diagnosis， treatment， and prognostic assessment.

Research Progress in Spatial Transcriptomics Technology for Liver Disease Research
Lingfei WAN, Wenting PAN, Yuting YONG, Yuanshuai LI, yue ZHAO, Xinlong YAN
2025, 15(4): 645-654. DOI: 10.19586/j.2095-2341.2025.0017
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The occurrence and development of liver diseases are finely regulated by various cell types and their spatial organization patterns. Spatial transcriptomics （ST） can achieve spatial localization of gene expression at the tissue slice level and has become an important technique for analyzing liver tissue to reveal dynamic changes in the disease microenvironment. The review summarized the research progress of ST technology in various liver diseases， such as alcoholic fatty liver disease， liver fibrosis， and hepatocellular carcinoma， sorted out the applications in revealing tissue spatial heterogeneity， intercellular interactions， and dynamic changes， and analyzed the current bottlenecks and future development directions of the technology. The aim is to provide data foundation and theoretical support for biomarker discovery， targeted therapeutic design，and personalized treatment strategies for early diagnosis and intervention of liver diseases.

Research Progress on the Mechanisms of Mercury Toxicity and Novel Prevention and Control Strategies
Yujie WANG, Pengyi YANG, Yang LI, Yiyang LI, Zhongqiang LI, Hongshu SUI, Zengshuo MAN
2025, 15(4): 655-660. DOI: 10.19586/j.2095-2341.2024.0169
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Mercury， as a global pollutant， represents a major research focus on public health due to its toxicity mechanisms and the development of prevention and control strategies. It mediates multi-organ toxicity through diverse pathways， including oxidative stress， epigenetic modifications， and mitochondrial dysfunction， with specific detrimental effects notably documented in the nervous， reproductive， and cardiovascular systems. Research has demonstrated that mercury exposure induces neuronal microtubule depolymerization， testicular fibrosis， and vascular endothelial dysfunction， and exacerbates multi-organ damage by activating inflammatory pathways such as NF-κB. Regarding prevention and treatment strategies， nanoparticle-based drug delivery systems and cellular adsorption techniques have significantly improved the targeting efficiency of chelation therapy. Concurrently， traditional Chinese medicine （TCM） formulations exhibit unique advantages in mitigating treatment-associated side effects. This review summarized the toxicity of mercury to humans and its damage mechanisms across various organ systems， along with current control measures. Future research directions should prioritize integrating multi-omics technologies to elucidate the complex networks underlying mercury toxicity and developing precision interventions based on epigenetic regulation， thereby providing a stronger theoretical foundation for the prevention and treatment of mercury poisoning.

Research Progress on Neurodevelopmental Toxicity in Offspring of PFASs Exposure
Lixin XIONG, Yujie WANG, Zhongle LI, Enming ZHONG, Shouyang YU, Yuxin JIANG, Hongshu SUI, Ronghua LIN
2025, 15(4): 661-667. DOI: 10.19586/j.2095-2341.2025.0023
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Per and polyfluoroalkyl substances （PFASs）， a class of highly persistent environmental pollutants， have become a research hotspot in recent years due to their neurodevelopmental toxicity in offspring mediated by the placenta. This review systematically elaborated on the molecular mechanisms by which PFASs affect the neurodevelopment of offspring through multiple pathways， including interfering with thyroid hormone balance， the expression of neurotrophic factors， the function of neural cells， and epigenetic regulation. It also assessed the limitations of existing intervention strategie， aiming to provide a theoretical framework for revealing the transgenerational toxicity mechanism of PFASs and reference for formulating policies to prevent and control maternal and infant exposure.

The Effect of SP140 Protein on the Expression Regulation of Several Tumor or Infection Related Genes and Some Immune Diseases
Zihan DONG, Jinhui SHAO
2025, 15(4): 668-674. DOI: 10.19586/j.2095-2341.2024.0170
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Human SP140， a leukocyte-specific nuclear protein inhibited by GSK761， belongs to the speck-like protein （SP） family and is predominantly expressed in immune cells. Current research indicates that SP140 activates gene transcription， regulates gene expression， and plays a significant role in immune modulation. This review systematically summarized the fundamental molecular characteristics of SP140， its role in the regulation of gene expression， and its involvement in immune responses. Furthermore， it detailed the association of SP140 with numerous diseases， primarily immune-related disorders， including primary biliary cholangitis （formerly cirrhosis）， multiple sclerosis， Crohn's disease， chronic lymphocytic leukemia， acquired immune deficiency syndrome（AIDS）， in order to enhance the understanding of the pathogenesis of these diseases and potentially contributing to improvements in their diagnosis and treatment.

Physiological and Photosynthetic Responses of Brassica pekinensis Seedlings to Salt Stress Alleviation by Chitosan Oligosaccharide
Lifeng E, Yani LI, Yu XIE, Jianhua QUAN, Xiubin CHEN, Jun HUA, Xuemei ZHAO, Wenqin ZHAO
2025, 15(4): 675-682. DOI: 10.19586/j.2095-2341.2025.0066
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To investigate the regulatory effects of chitooligosaccharides （COS） on physiological characteristics of Brassica pekinensis seedlings under salt stress， the cultivar "Cold-tolerant Golden Empress" was used. Seedlings were foliar sprayed with 0 （CK）， 50， 100， 150， or 200 mg·L^-1 COS for 4 days and then subjected to salt stress with 150 mmol·L^-1 NaCl. The results showed that the 100 mg·L^-1 COS treatment significantly increased plant height and dry weight per plant under NaCl stress， increasing by 14.44% and 88.06%， respectively. It also significantly enhanced the activities of antioxidant enzymes peroxidase （POD） and catalase （CAT） by 16.41% and 33.77%， respectively， while significantly reducing malondialdehyde （MDA） content. Furthermore， the net photosynthetic rate （Pn）， transpiration rate （Tr）， and stomatal conductance （Gs） of the leaves were significantly increased by 28.26%， 41.67%， and 15.90%， respectively， whereas intercellular CO₂concentration （Ci） was significantly reduced. Proline content also increased significantly. These results indicated that 100 mg·L^-1 COS effectively alleviates salt stress-induced damage in Brassica pekinensis seedlings by enhancing antioxidant defense and photosynthetic efficiency， providing a theoretical basis and technical support for stress-resistant cultivation of highland summer vegetables.

Pollution Status and Ecological Risk Assessment of Organophosphate Insecticides Along the Inflow Routes to the Taihu Lake Basin
Xin GENG, Weiwei FENG, Guanghua MAO, Yao CHEN, Xiangyang WU
2025, 15(4): 683-692. DOI: 10.19586/j.2095-2341.2025.0022
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To study the pollution status and ecological safety of organophosphate insecticides （OPs） along the inflow routes to Taihu Lake Basin， water， suspended particulates （SPM） and sediment samples were collected from the watershed during the abundant and dry seasons， and the GC method was used to detect the types and contents of OPs， analyze the seasonal distribution patterns， partition coefficients， and potential sources of contamination， and then assess the ecological risks. Phoxim （PHO）， dichlorvos （DDVP）， dimethoate （DMT） and fenitrothion （MEP） were detected in water， SPM and sediment samples. In the water samples， the concentrations of PHO， DMT and DDVP were relatively high and they accounted for a larger proportion， while MEP was relatively less. The K_d1 and K_d2 of Σ₄OPs were 0.44~1.32 L·g^-1 and 2.97~6.88 L·g^-1 respectively during the abundant water period， and 0.34~1.40 L·g^-1 and 2.85~7.43 L·g^-1 respectively during the depleted water period， which indicated that the distributions of the OPs were relatively stable in the water samples， SPM samples and the sediments. To sum up， the water along the the Taihu Lake basin has been polluted by organophosphorus pesticides. The discharge of industrial and agricultural wastewater may be the main source of OPs along the Taihu Lake basin， and the two OPs pollutants DMT and MEP have potential ecological risks to the main rivers along the Tongji River in Zhenjiang City.

Study on the Mechanism of Bubble Generation and Inhibition Method During Digital PCR Amplification Process
Yanyan JIA, Luyang DUANMU
2025, 15(4): 693-701. DOI: 10.19586/j.2095-2341.2025.0010
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The emergence of digital PCR （dPCR） technology has significantly promoted the development of nucleic acid detection. However， the problems of droplets moving， breaking and generating bubbles in microfluidic chips have limited the development of digital PCR technology and it is of great significance to solve the existing problems. The article constructed a device and method for droplet PCR in microfluidic chips based on constant pressure regulation analyed the existence principle of bubbles， and the bubbles in the chip were simulated and verified. The results showed that adjusting the pressure in the chip through the air source device could achieve bubble-free generation of droplets and PCR amplification. The quantitative detection of human epidermal growth factor receptor （EGFR） exon 18 gene was carried out by using the device and the chip. Within 3 minutes， 20 μL of the sample was distributed into about 50 000 water-in-oil droplets， which were tightly arranged without bubbles in the chip. The results showed that the experimental detection signal had a good linear relationship with DNA concentration in the range of 10¹~10⁵ copies·μL^-1 （R²=0.999）. Compared with current research on bubble removal， dPCR technology avoids the fragmentation and fusion of micro-droplets and can standard the results， which has broad application prospects in nucleic acid quantitative detection.

Analysis of Molecular Characteristics of Megakaryocytes Between Embryonic Aorta-gonad-mesonephros and Adult Bone Marrow
Yifei CAI, Yezi MA, Meijuan XIA, Cuicui LIU, Hongtao WANG, Jiaxi ZHOU
2025, 15(4): 702-710. DOI: 10.19586/j.2095-2341.2025.0009
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Megakaryocytes are a type of cell primarily present in adult bone marrow， with the main function of producing platelets. In addition， adult bone marrow megakaryocytes also play a role in maintaining the homeostasis of hematopoietic stem cells （HSCs）. Recent studies have revealed that megakaryocytes also exist in the aorta-gonad-mesonephros （AGM） region and are involved in regulating the generation and maturation of hematopoietic stem cell precursors （pre-HSCs）. The AGM region is the earliest site for HSC production， while the bone marrow is the main site for HSC colonization and hematopoiesis in adulthood. However， it remains unclear whether megakaryocytes regulate HSC development and function through distinct molecular mechanisms at these two sites. Based on recently reported single-cell transcriptome data of megakaryocytes from the AGM region in mouse embryos and from adult bone marrow， we performed a comparative analysis of their overall molecular characteristics and the molecular features associated with regulating HSC biological processes. The results showed significant differences in molecular characteristics between AGM region megakaryocytes and adult bone marrow megakaryocytes in mice. Specifically， AGM region megakaryocytes highly expressed molecular features related to cell proliferation， circulatory system development， stem cell development， and differentiation； in contrast， adult bone marrow megakaryocytes highly expressed molecular features associated with immune response， response to external stimuli， hemostasis and coagulation， and cell communication. Furthermore， AGM region megakaryocytes and adult bone marrow megakaryocytes highly expressed different HSC regulation-related molecules. These findings suggested that megakaryocytes in the AGM region and adult bone marrow may regulate HSC development and function through distinct functional molecules， providing a theoretical basis for investigating the functional differences between embryonic and adult megakaryocytes and their underlying molecular mechanisms.

Mechanism of miR-93 Regulating Immune-mediated Traumatic Brain Injury Through Activation of PI3K/AKT Pathway
Xiaoyu JIANG, Yi CHEN, Jingjing NI
2025, 15(4): 711-719. DOI: 10.19586/j.2095-2341.2025.0027
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This study aimed to investigate the mechanism by which miR-93 promotes traumatic brain injury （TBI） repair through regulating macrophage M2 polarization and neurovascular tissue regeneration via the PI3K/AKT signaling pathway.Female C57BL/6 mice were randomly assigned to four groups： sham-operated （Sham）， TBI model （TBI）， TBI + miR-93 inhibitor （TBI+inhibitor）， and TBI + miR-93 agonist （TBI+agonist）. Relative mRNA expression levels of M1 macrophage marker CD80 and M2 macrophage marker CD206 were assessed by quantitative real-time PCR （qPCR）. Concentrations of inflammatory cytokines TNF-α， IL-12， TGF-β， and IL-13 were measured using enzyme-linked immunosorbent assay （ELISA）. Immunofluorescence staining for vascular endothelial growth factor （VEGF） was performed to evaluate angiogenesis post-TBI. Western blotting was used to determine the relative protein expression levels of phosphorylated PI3K （p-PI3K） and phosphorylated AKT （p-AKT）.Compared to the TBI group， the TBI+agonist group exhibited enhanced M2 macrophage polarization， reduced expression of pro-inflammatory cytokines TNF-α and IL-12， and increased expression of anti-inflammatory cytokines TGF-β and IL-13. Furthermore， this group showed a lower modified Neurological Severity Score （mNSS）， indicative of improved neurological function， enhanced VEGF expression reflecting active angiogenesis， and elevated levels of p-PI3K and p-AKT proteins.These results suggested that miR-93 expression is compensatorily upregulated following TBI. miR-93 promotes neurovascular repair after injury by modulating macrophage polarization towards the M2 phenotype， likely via activation of the PI3K/AKT signaling pathway.

Echinoside Inhibits IL-1β-induced Chondrocytes Iron Death Through Nrf2/HO-1 Pathway
Chuancai LIANG, Bo QIU
2025, 15(4): 720-725. DOI: 10.19586/j.2095-2341.2025.0043
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To investigate the protective mechanism of echinacoside （ECH） in interleukin-1β （IL-1β）-induced chondrocyte iron death. Wistar mammary rat chondrocytes were extracted， and the experiment was divided into 5 groups： control group， IL-1β group， ECH+IL-1β group， ECH+IL-1β+Brusatol （Nrf2 inhibitor） group and ECH+IL-1β+ZnPP （HO-1 inhibitor） group. Quantitative PCR （qPCR） and Western Blot were used to detect mRNA and protein expression nuclear factor E2-related factor 2 （Nrf2）， heme oxygenase-1 （HO-1）， esteroyl coenzyme A synthase long-chain family member 4 （ACSL4） and glutathione peroxidase 4 （GPX4）， respectively. The results showed that the mRNA and protein expression of Nrf2， HO-1 and GPX4 were significantly reduced in the IL-1β group， and the mRNA and protein expression of ACSL4 were significantly enhanced compared with the control group. The mRNA and protein expression of cartilage Nrf2， HO-1 and GPX4 were significantly decreased and that of ACSL4 was significantly enhanced by the addition of the Nrf2 inhibitor Brusatol or the HO-1 inhibitor ZnPP in the ECH group. The results of Fe²⁺ Assay Kit showed that the chondrocyte Fe²⁺ content was significantly higher in the IL-1β group compared to the control group. The chondrocyte Fe²⁺ content was significantly lower in the ECH group （ECH+IL-1β） compared to the IL-1β group， while it was significantly higher in the ECH group after the addition of the Nrf2 inhibitor Brusatol or the HO-1 inhibitor ZnPP. The results suggest that echinoside can significantly improve IL-1β-induced chondrocyte iron death through Nrf2/HO-1 pathway.

Role and Mechanisms of DBNDD1 in Colorectal Cancer Development
Xiaoya LIU, Shuomin ZHANG, Peng ZHENG, Rui MA, Chaojun ZHANG
2025, 15(4): 726-734. DOI: 10.19586/j.2095-2341.2025.0033
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The malignant progression of colorectal cancer （CRC） is closely associated with abnormal activation of critical oncogenes. Analysis of the cancer genome atlas （TCGA） omics data revealed that DBNDD1 is significantly upregulated in CRC tissues， with its overexpression demonstrating a significant correlation with shorter overall patient survival. This study aimed to investigate the expression characteristics， biological functions， and clinical significance of DBNDD1 in CRC. The research integrated omics analysis with experimental validation， evaluating the prognostic relevance of DBNDD1 expression through the TCGA database， assessing migration， invasion capabilities （Transwell assay， wound healing assay）， and apoptosis levels （flow cytometry） in HCT116 cells following DBNDD1 knockdown using siRNA， and monitoring tumor growth and apoptosis changes （Ki-67 immunohistochemistry， TUNEL assay） in nude mouse xenograft models. Gene set enrichment analysis （GSEA） was further employed to explore associated signaling pathways and immune microenvironment regulatory mechanisms. The experimental results demonstrated that DBNDD1 knockdown suppressed migration and invasion capabilities of HCT116 cells while promoting apoptosis. Mechanistic analysis revealed significant enrichment of NF-κB and TNF signaling pathways in DBNDD1-overexpressing samples， along with close associations with immunosuppressive microenvironment features， including increased infiltration of M2 macrophages and regulatory T cells. These findings provide novel research directions for the development of clinical molecular biomarkers and targeted therapeutic strategies in colorectal cancer.

Construction of a Breast Cancer Prognostic Model Based on Nicotine Metabolism Gene Signatures
Ziyi ZHANG-HUANG, Lisha HUANG, Yanqi LI, Chenlu XIONG, Ying YU, Fei XIE
2025, 15(4): 735-742. DOI: 10.19586/j.2095-2341.2025.0013
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Breast cancer， as the most prevalent malignancy among women globally， exhibits significant prognostic heterogeneity. Transcriptomic data from breast cancer patients in the cancer genome atlas （TCGA） database were analyzed through differential expression and correlation analyses to identify nicotine metabolism-related differentially expressed genes. A prognostic model was constructed using univariate Cox regression and Lasso-Cox regression analyses， and its performance was evaluated using Kaplan-Meier survival curves， ROC curves， calibration curves， and clinical factor-integrated nomograms. The GSE42568 dataset from the gene expression omnibus （GEO） was employed as an external validation cohort for model verification， and further condueted immune infiltration and drug sensitivity assessments. The results showed that the model was composed of 10 nicotine metabolism-related genes， which could be validated as an independent prognostic factor for breast cancer. Immune infiltration and drug sensitivity analyses revealed significant differences in immune cell infiltration and drug responsiveness between high- and low-risk groups. This risk prognostic model can independently predict breast cancer outcomes and provide novel molecular markers and potential therapeutic targets for prognostic evaluation and personalized treatment of breast cancer.

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