Current Biotechnology

Research Progress of Iron Signaling and its Role in Plant-pathogen Interaction
Wenxuan PU, Xi DAI, Jiani YUE, Xiuxia FU, Na SONG, Wei LI, Yu PENG
2025, 15(1): 1-10. DOI: 10.19586/j.2095-2341.2024.0121
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Iron is an essential trace element for plants and plant pathogens. Although the iron content is rich in soil， it couldn't be absorbed directly by plants due to the iron existing with the form of insoluble compounds. To supply growth and development， the plants， with long-term evolution process， have formed two iron absorption systems based on the reduction mechanism and chelation mechanism， and iron transport systems. Here， we reviewed the progress of the two strategies and molecular mechanisms of iron uptake in plants， the transport of iron in plant cells and the pathways in response to iron signaling in plants， the influence of iron on plant immune responses， the effect on pathogenicity of pathogens， and the functional mechanism of iron in plant-pathogen interaction. The aim of this review was providing reference for the study of iron signaling pathways in plant and the function of iron in plant-pathogen interaction，providing new ideas for crop cultivation.

An Overview on Fiber Improvement Transgenic Cotton Studies
Xiaoyuan WANG, Xi BAI, Jiansheng WANG, Hongzhi CUI
2025, 15(1): 11-18. DOI: 10.19586/j.2095-2341.2024.0095
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As a natural fiber， cotton is the most important raw material for the textile industry and one of the most important cash crops in the world. With the pursuit of high-quality cotton fiber， increasing yield and improving cotton fiber quality are important goals of cotton breeding in China. The discovery of the key factors or related functional genes regulating the development of cotton fiber is not only helpful to understand the mechanism of cotton fiber development， but also provides new ideas for cultivating new varieties of high-quality cotton fiber. In this paper， we summarized the important role of related factors in fiber development and regulation from five aspects： fiber elongation， cytoskeletal structural proteins， plant hormones， carbohydrate metabolism and lignin metabolism， as well as some advances in the research of fiber-improved transgenic cotton. We hope to provide reference for further studies of the molecular mechanisms regulating cotton fiber development and related research on improving cotton fiber quality.

Research Progress on Metabolites of Marine Streptomyces Under the Strategy of Activating Silencing Genes
Boxuan WANG, Jiaxin CHEN, Jiajun CAI, Kunxian LI, Jiachun LIN, Lingzhi TANG, Xuan HONG
2025, 15(1): 19-26. DOI: 10.19586/j.2095-2341.2024.0141
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Marine Streptomyces are the source of the valuable lead compounds with medicinal biological activity due to their complex morphological differentiation cycle， which often accompanied by complex physiological changes. But most of the biosynthetic gene clusters of secondary metabolites in marine Streptomyces are “silent”. The article reviewed the methods for predicting the expression potential of “silent genes” and summarized the means of activating “silent genes”， including changing culture conditions， chemically inducing gene cluster expression and biological mediation， in order to provide new avenues for the mining of secondary metabolites in marine Streptomyces.

The Role of the Global Regulatory Factor LaeA in Filamentous Fungi: Mechanisms and Applications
Xianchao MENG, Bo HU, Zhe NING, Yuanyuan SUN, Haobing YU, Xiaoyu LIU
2025, 15(1): 27-34. DOI: 10.19586/j.2095-2341.2024.0105
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Loss of aflR expression-A （LaeA） has been recognized as a crucial global regulatory factor in filamentous fungi， orchestrating the intricate balance of gene expression to govern either the developmental trajectory of strains or the synthesis and diversification of secondary metabolites. Recent advancements in bioinformatics analysis have unveiled numerous novel regulatory roles of LaeA. This review focused on elucidating the mechanisms and regulatory functions of LaeA， encompassing its structural-functional attributes and its modulation of fungal growth and secondary metabolite biosynthesis. Simultaneously， it explored the potential applications of LaeA in the fields such as drug development， agriculture， and industrial production， in order to provide reference and help in improving the yield and activity of secondary metabolites of filamentous fungi，and developing new marine drugs.

Application and Prospect of CRISPR/Cas9 Gene Editing Technology in Disease Treatment
Yiyang LI, Zhizheng ZHOU, Shufei WANG, Boya LIU, Yufei LIU, Xiaoyan LI, Hongshu SUI, Dongwei LIU
2025, 15(1): 35-42. DOI: 10.19586/j.2095-2341.2024.0148
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The clustered regularly interspaced short palindromic repeats associated proteins （CRISPR) system is an acquired immune system of prokaryotes. The CRISPR/Cas9 system， developed based on the bacterial immune system CRISPR， is changing biology and basic medical research， and is one of the most efficient， simplest and cost-effective gene editing and modification technologies available. However， there is currently a lack of strategies for effectively delivering CRISPR systems to diseased cells in vivo， and non-viral vectors with target recognition capabilities may be the focus of future research， with pathological and physiological changes caused by disease onset promising as identifying factors for targeted delivery or gene editing targets. This article provided an overview of existing gene editing tools and the advantages of the CRISPR/Cas9 system， summarized the application of CRISPR/Cas9 in the field of therapy， and discussed the problems and challenges encountered in CRISPR/Cas9-mediated therapy， in order to promote the advancement of CRISPR/Cas9 therapeutical technology and provide new perspectives for treating other complex diseases.

Research Progress of Antibody Drug Screening Technology
Sijia JI, Xiaoxue ZHANG, Lili ZHAI, Jing DONG, Yanyi LI, Liang CHANG
2025, 15(1): 43-49. DOI: 10.19586/j.2095-2341.2024.0149
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Antibody drugs have gone through three stages： polyclonal antibody， monoclonal antibody and genetically engineered antibody. With the continuous deepening of human research on antibody drugs， a variety of antibody screening technologies have emerged， such as polyclonal antibody technology， hybridoma antibody technology， antibody library display technology and transgenic mouse technology. Antibody drugs have a very large development prospect. The article summarized the characteristics of various antibody drug screening technologies， and discussed the challenges faced by antibody drug research and development， in order to provide reference for the subsequent development of antibody technology.

Research and Application of Protein Purification Technology Based on Different Ligands
Xiaomin PANG, Yanyi LI
2025, 15(1): 50-57. DOI: 10.19586/j.2095-2341.2024.0153
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As the material basis of life， proteins are the main carriers of life activities. For downstream processes， various impurities in complex protein samples can be removed more safely， economically， and efficiently for the continuous development of new ligand purification technologies. The research status， innovative applications and development prospects of medium materials with different ligands （including hydrophobic ligands， ion exchange ligands， mixed-mode ligands and short peptide biomimetic ligands） and protein separation technologies （including polymer grafting technology and continuous column chromatography technology） were summarized， which with the hope of obtaining effective resources in the protein purification process and providing a reference for the development of downstream purification technologies.

Research Progress on Measuring Technology of Bio-enzyme Activity
Xiaoni HOU, Mingdong LIU, Hao LYU, Deping YE, Lixia MA, Lihua ZHOU
2025, 15(1): 58-66. DOI: 10.19586/j.2095-2341.2024.0158
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Biological enzymes play significant roles in the agriculture， industrial production， medicine and environment protection. Activity is the main performance parameter of bio-enzymes， and directly affects the quality and application of biological enzymes. With the development of science and technology， the detection methods of biological enzyme activity have also produced many new technologies and new methods. This paper reviewed the mainstream test techniques and methods of biological enzyme activity at home and abroad， including spectroscopy， electrochemical， chromatography， mass spectrometry， surface plasmon resonance， surface enhanced Raman spectroscopy. We also introduced the basic principles， applicable conditions and advantages and disadvantages of different methods. Finally， some suggestions were put forward for the accuracy and reliability of the test results of biological enzyme activity. The paper was expected to provide guidance on biological enzyme activity detection， method selection， quality control and product application.

Mechanism of Lignin Content in Root System Affecting Salt Tolerance in Maize
Qingyun ZHANG, Lei MA, Hua XU, Lian JIN, Junjie ZOU, Baobao WANG, Quanjia CHEN, Miaoyun XU
2025, 15(1): 67-77. DOI: 10.19586/j.2095-2341.2024.0107
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Soil salinity is a major constraint on global agricultural production， posing a severe threat to agriculture sustainable development and food security. Maize （Zea mays L.） is one of the three major crops in China， while saline-alkali land represents a crucial reserve of arable land resources. Lignin， as a principal structural component of plant cell walls， makes it significant to study the accumulation of lignin in maize and the thickening of cell walls in response to high salinity. This study selected salt-tolerant maize inbred lines （Zhongke4M， Zheng58） and salt-sensitive maize inbred lines （PH4CV， Chang7-2） as subjects. Using a water control and a treatment of 200 mmol·L^-1 NaCl， the morphological changes and cytological characteristics of maize roots under different salt concentrations were analyzed. The differences in enzyme activity， lignin content， and gene expression were also examined. Toluidine blue staining results indicated that the reduction in the area of the root cortex and endodermis under salt stress was significantly lower in the salt-tolerant inbred lines Zhongke4M and Zheng58 compared to the salt-sensitive inbred lines PH4CV and Chang7-2. Furthermore， fluorescence microscopy observations showed that the degree of lignification in the salt-tolerant inbred lines remained stable or increased under salt stress， whereas the salt-sensitive inbred lines exhibited a decrease in lignification. The results demonstrated that the lignin content in the salt-tolerant inbred lines Zhongke4M and Zheng58 remained stable under salt stress， while it significantly decreased in the salt-sensitive inbred lines. Enzyme activity analysis revealed that under salt stress， the activity of phenylalanine ammonia-lyase （PAL） and cinnamate-4-hydroxylase （CAD） decreased in the salt-sensitive inbred lines， while the activity of cinnamate-4-hydroxylase （C4H） increased in the salt-tolerant inbred lines. RNA-seq analysis identified three genes related to lignin biosynthesis， whose expression levels varied among different maize varieties. This study provided new insights into the mechanisms by which maize responds to salt stress through the regulation of lignin accumulation and cell wall structure， contributing to the understanding of maize salt tolerance mechanisms.

Comparison of Endogenous Genes in Maize Based on Digital PCR
Xin QI, Xinran LI, Yaning GUO, Dan WANG, Kai LI, Qiong WU, Liang LI
2025, 15(1): 78-85. DOI: 10.19586/j.2095-2341.2023.0165
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With the increasing variety and planting area of transgenic maize， the demand for qualitative and quantitative detection of transgenic maize-related products is increasingly urgent. The selection of stable genetic and specific endogenous genes is of great significance for the accuracy and consistency of transgenic maize identification results. In this study， digital PCR technology was used to screen and compare nine endogenous genes in maize， including hmg， adh1-1， adh1-2， ivrI-1， ivrI-2， zSSIIb-1， zSSIIb-2， zein-1 and zein-2. By optimizing the annealing temperature of the above nine kinds of endogenous genes， the optimal annealing temperature for each gene was found. Taking transgenic maize MON810 as the research object， the above endogenous genes were used to determine the actual samples. The results showed that adh1-1 and adh1-2 could be used for copy number detection of actual samples， which would provide reference for endogenous gene selection and related quantitative detection of transgenic maize in the future， and provid a strong technical support for the management of transgenic crops in China.

Establishment of Detection Method Based on TaqMan Real-time Fluorescence Quantitative PCR Technology for OsWx-edited Rice
Guang HU, Zhi WANG, Wei FU, Yuting SHI, Shanshan CHEN, Liang LUO, Shuang WEI
2025, 15(1): 86-92. DOI: 10.19586/j.2095-2341.2024.0154
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Gene editing technology currently has become an important molecular tool， which can selectively modify genome sequences and has wide applications in gene function analysis and crop genetic breeding. Due to the rapid development of gene editing technology， gene editing techniques can perform almost seamless editing on receptor organisms， which increases the difficulty of detecting the gene editing product. Therefore， the establishment of detection methods for gene editing products is conducive to strengthening market supervision of the gene editing product. This study was based on TaqMan real-time fluorescence quantitative PCR （TaqMan-qPCR） technology for detection to design specific primers and probes for the editing site of OsWx gene by developed with CRISPR/Cas9. In this study， the detection system had good specificity and sensitivity， and could effectively distinguish OsWx single base mutants from wild-type rice.

The Impact of Enzyme Treatment on Yeast Fermentation of Tobacco
Xiaomin LIU, Ting LU, Yong LI, Meng WANG, Baokun ZHU, Wei ZHANG
2025, 15(1): 93-101. DOI: 10.19586/j.2095-2341.2024.0093
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The research aimed to study the influence of different enzyme preparations on fermentation of tobacco. In this experiment， tobacco produced in Yunnan Province were used as materials. Different enzyme preparations were applied to enzymatically treat tobacco， followed by fermentation using wine yeast. The aim was to prepare tobacco products with richer and fuller flavor. The effects of different enzyme treatments on the conventional chemical composition， volatile flavor substances， and sensory quality of tobacco were analyzed. The results showed that， there was no significant change in moisture content and nitrogen content of tobacco after enzymatic treatment， with nitrogen content ranging from approximately 0.98% to 1.18%. The group treated with 100 U·g^-1 flavor protease exhibited the highest content of soluble total sugars （12.11%） and reducing sugars （5.93%）. The total amount of volatile flavor substances in all groups of tobacco increased， with the highest being 180.029 μg·g^-1. Moreover， the content of characteristic flavor substances in each group of tobacco， such as neophytadiene， phenylethanol， solanone， and megastigmatrienone， also increased. The sensory quality of tobacco improved after enzymatic treatment. Comprehensive analysis revealed that enzymatic treatment using 70 U·g^-1 flavor protease and 50 U·g^-1 α-amylase resulted in a more balanced chemical composition， abundant aroma components， and better sensory indicators of tobacco. Therefore， enzymatic fermentation can improve the quality of tobacco leaves， providing a basis for further commercial development of tobacco leaves.

Determination and Analysis of Nutritional Components, Active Substances and Heavy Metal Content in Artemisia indica
Ningjie LI, Bingnian JING, Wei WANG, Yuqing LIU, Xiaoyang XIE, Genlai DONG, Xuefang WANG, Lei WEI
2025, 15(1): 102-109. DOI: 10.19586/j.2095-2341.2024.0124
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Artemisia indica is used as a food ingredient in the folk， but there is a lack of systematic measurement and analysis of its nutritional components， active substances and heavy metal content. In this study， national standards and literature methods were used to determine the nutritional components， mineral elements， vitamins， amino acid composition， active substances and heavy metal content. The results showed that the descending order of nutritional components in Artemisia indica was： protein （31.62%）， carbohydrate （29.86%）， fiber （22.57%）， total sugar （20.94%）， ash （8.76%） and fat （7.42%）. It was rich in VK₁， VB₁， VB₂， VB₁₂ and folic acid， especially the content of VB₂ was as high as 1 mg·100 g^-1. There were many mineral elements such as Na， K， Ca， Zn， Mg， Mn， Fe， Cu and Se， of which K content was the highest， up to 28 747.71 mg·kg^-1. In terms of amino acid composition， the protein was rich in various amino acids， including 8 kinds of essential amino acids. The content of essential amino acids accounted for 40.13% of the total amino acids， and the functional amino acids accounted for 56.7%. The main active ingredients were triterpenoids and polysaccharides， with contents of 5.38% and 2.91%， respectively. The contents of Hg， Pb， Cr and As were all below the limits of national food safety standards except Cd. Therefore， Artemisia indica is a food resource with high protein， high fiber， low fat， rich mineral elements types， high content B-group vitamins， and active substances such as triterpenoids and polysaccharides. This results will provide a theoretical basis for further research and development of Artemisia indica.

Research on the Synthesis of Cinnamyl Alcohol Glycosylated Derivatives Based on Immobilized Enzyme Method
Xinran CHENG, Weiyong ZONG, Wenfang DOU
2025, 15(1): 110-118. DOI: 10.19586/j.2095-2341.2024.0157
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Rosin is a phenylpropanoid glycoside compound with various biological activities. The traditional way of producing rosin is through plant extraction， but natural resources are limited. This article utilized the coupling catalysis of glycosyltransferase from Arabidopsis thaliana and sucrose synthase from soybean to immobilize two enzymes with His tags， converting cinnamyl alcohol into a complex and regenerating uridine diphosphate glucose （UDPG）. Two enzymes were fixed separately， and it was determined that 5 g of UGT73C5 crude enzyme and 6 g of GmSUS crude enzyme could be fixed with 2 g of resin. The optimal conditions for the immobilized coupling catalytic reaction were optimized， and the coupling immobilized enzyme was tested to be reproducible and stable for seven batches. Finally， a batch feeding strategy was adopted， and the yield of the complex plug reached 15.82 mmol·L^-1 with a conversion rate of 97.5%. After the reaction， high-performance liquid chromatography（HPLC）was performed， and it was inferred that the reaction product is rosin. The strategy of immobilizing enzymes will open up new prospects for the development of double enzyme pairs to form rosin， and reduce the cost of free enzymes in large-scale industrial applications.

Association Analysis Between EPAS1 Gene Polymorphism and Blood Physiological Indicators in Two Different Altitudes of Sheep in Xinjiang
Jiong DONG, Shifeng WANG, Kaidiriye YUSUPU·, Jianli ZHAN, Abudureheman TUERXUN·
2025, 15(1): 119-126. DOI: 10.19586/j.2095-2341.2024.0137
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To explore the relationship between EPAS1 gene polymorphisms and hemoglobin indices in hypoxia adaptation among local Ovis aries populations in Xinjiang at varying altitudes， exons 9 and 16 of the EPAS1 gene were PCR-amplified and sequenced in 79 Tashkurgan sheep （4 200 m altitude） and 74 Duolang sheep （1 200 m altitude）. 7 SNP loci were identified， gene frequencies and differences in hemoglobin indices were analyzed. The results revealed that the hemoglobin content in Tashkurgan sheep was significantly higher than in Duolang sheep， suggesting that Tashkurgan sheep adapt to high-altitude hypoxia by increasing hemoglobin levels. Additionally， exons 9 and 16 of the EPAS1 gene were relatively conserved， with amino acid-level variation being significantly lower than nucleotide-level variation. Duolang sheep exhibited greater genetic diversity at the same polymorphic loci， indicating a negative correlation between altitude and polymorphic information content. These findings provide insights into the molecular mechanisms underlying Tashkurgan sheep's adaptation to high-altitude hypoxia and changes in blood physiological indices.

Microbial Contamination and Antibiotic Resistance Characteristics in Raw Milk from Pastures in Northern Fujian Province
Shuanghui ZHOU
2025, 15(1): 127-134. DOI: 10.19586/j.2095-2341.2024.0164
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To investigate the microbial contamination and antibiotic resistance characteristics of pathogenic bacteria isolated from raw milk in pastures of northern Fujian Province. From December 2023 to June 2024， 46 bulk tank raw milk samples were collected from 14 pastures in northern Fujian. The samples were analyzed for aerobic plate count， coliform count， and screened for contamination by five pathogenic bacteria： Staphylococcus aureus， Salmonella spp.， Listeria monocytogenes， Cronobacter spp.， and Klebsiella spp. Additionally， antibiotic susceptibility tests were conducted on the isolated strains of Staphylococcus aureus， Klebsiella spp.， and Escherichia coli. The average total bacterial count in the 46 raw milk samples was 5 500 CFU·mL^-1， and the average coliform count was 39 CFU·mL^-1. The overall antibiotic resistance rate of the isolated pathogenic bacteria was 63.5%， with resistance rates of 38.3% for Staphylococcus aureus， 96.6% for Klebsiella spp.， and 84.6% for Escherichia coli. The highest resistance was observed to β-lactam antibiotics， with 93.2% of the pathogenic bacteria exhibiting multidrug resistance. The microbial contamination risk in the raw milk from northern Fujian pastures is low， meeting the “premium milk” standard. However， there is a significant issue with antibiotic resistance among the pathogenic bacteria， particularly to β-lactam antibiotics， with a high risk of multidrug resistance.

Construction and Expression of Lentiviral Vector for Overexpression of WIPI2 Gene
Yan LUO, Qi HONG, Leyi LIANG, Shiyue YU, Qiongxia CHEN, Xiaofang HAN, Hongyan ZHEN
2025, 15(1): 135-141. DOI: 10.19586/j.2095-2341.2024.0098
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WD repeat domain， phosphoinositide interacting 2 （WIPI2） is overexpressed in colorectal cancer （CRC）， suggesting that WIPI2 may be closely related to CRC progression， but the precise mechanism remain unclear. Therefore， the construction of a CRC cell model with stable overexpression of WIPI2 can provide a powerful experimental tool for in-depth study of the effect mechanism of WIPI2 in CRC and discovery of potential therapeutic targets for CRC. In this study， lentiviral vectors overexpressing WIPI2 were designed according to the human WIPI2 gene sequence and infected with CRC cells to evaluate the effect of the recombinant lentiviral vectors in enhancing the expression of WIPI2 protein. The sequence of the recombinant lentiviral vector was confirmed to be consistent with the target sequence of WIPI2 by gene sequencing， and the viral titer of the recombinant lentivirus was measured after viral packaging using 293T， and the titer of GL190-WIPI2 was 2.16×10⁸ TU·mL^-1； the expression level of WIPI2 protein in the overexpression of WIPI2 stably-transfected cell line was obviously up-regulated， and the lentiviral vector for overexpressing WIPI2 was successfully constructed and established. A lentiviral vector for WIPI2 overexpression was successfully constructed， and a stable transient CRC cell line overexpressing WIPI2 gene was established， which provides a cell model for studying the role of WIPI2 in CRC.

Pan Cancer Analysis Between ITGA7 Gene and Human Tumors
Sibin ZHEN, Yining CHE, Xiao WANG, Mian LI
2025, 15(1): 142-151. DOI: 10.19586/j.2095-2341.2024.0073
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This study aimed to explore the gene and protein expression levels of ITGA7 gene in human tumors and evaluate its clinical significance. The variation of ITGA7， its relationship with tumor prognosis， the correlation between gene expression and immune infiltration， and functional enrichment analysis were analyzed through various databases such as cBioPortal， GEPIA2， Timer2.0， STRING and Veen. It was found that increased expression of ITGA7 in tumor tissues such as liver cancer， renal clear cell carcinoma， and thyroid cancer was associated with poor prognosis. On the contrary， ITGA7 expression in tumor tissues such as endometrial cancer， prostate cancer， rectal cancer， breast cancer， and lung squamous cell carcinoma was found to be associated with poor prognosis. The expression of ITGA7 is low in pancreatic cancer， and its low expression is associated with poor prognosis of pancreatic cancer. The main mutation type of ITGA7 is missense mutation. In six types of tumors， including colon cancer， bladder cancer， and breast cancer， the expression of ITGA7 was positively correlated with the level of immune infiltration， while the expression in sarcomas was negatively correlated with the level of immune infiltration. Pan-cancer analysis showed that the expression of ITGA7 is related to the level of immune infiltration， gene mutations and tumor prognosis. This finding helps to deeply understand the potential role of ITGA7 in tumor occurrence and development from a clinical perspective， and provides a reference for clinical diagnosis and treatment.

Application of Recombinant Trypsin in Tissue Isolation of Primary Hamster Kidney Cells
Jian QIAO, Shuai ZENG, Yang ZHANG, Gelin XU
2025, 15(1): 152-157. DOI: 10.19586/j.2095-2341.2024.0074
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To explore the feasibility of using recombinant trypsin as a substitute for porcine derived trypsin in the preparation of attenuated Japanese encephalitis live vaccine， porcine derived trypsin， recombinant trypsin derived from Escherichia coli BL21 （DE3） expression system， and recombinant trypsin derived from Pichia pastoris （X-33） expression system were used to digest primary hamster kidney blocks at the same enzyme activity （total enzyme activity 315 200 U）， with the same digestion temperature and time in each group. The results showed that after digestion with three types of pancreatic enzymes， the cell dispersion effect and growth status were good， and there was no significant difference in the cell spreading rate on the third day of cultivation （P>0.05）. During the virus culture stage， there was no significant difference in virus titers among the three groups of virus supernatants （P>0.05）. The results showed that recombinant trypsin derived from E. coli BL21 （DE3） expression system and Pichia pastoris （X-33） expression system can replace porcine trypsin in the preparation of attenuated live Japanese encephalitis vaccine.

Pancreatic Cancer Risk Stratification and Prognostic Prediction Model Based on Aging-related Gene Characteristics
Xinxiong LI, Weixuan HONG, Tianshun FENG, Junwei FANG, Hu ZHAO, Chunhong XIAO, Meiping WANG
2025, 15(1): 158-169. DOI: 10.19586/j.2095-2341.2024.0027
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In order to identify aging-related long noncoding RNAs （lncRNAs） with prognostic significance in patients with pancreatic adenocarcinoma （PAAD）， a prognostic prediction model was constructed based on aging-related genes （ARG）. The transcriptome information of PAAD and normal samples were obtained from The Cancer Genome Atlas Database （TCGA） and Genotypic Tissue Expression Database （GTEX）， and correlation analysis screened out lncRNAs with significant correlations among ARG. The target lncRNA was identified through differential analysis， univariate regression， Lasso regression and multivariate regression， and a PAAD prognostic risk model was constructed. A total of 1 109 aging-related lncRNAs were identified， and 9 lncRNAs were screened to construct a risk score prognostic model， including AC245041.2， AC244153.1， AC091057.1， MIR3142HG， AL137779.2， AC145207.5， TDRKH-AS1， AC068620.2 and AC127024.6. The receiver operating curve （ROC） curve and AUC （the area under the curve） reached 0.798. The patients were divided into high-risk group and low-risk group according to the median risk. Kaplan-Meier analysis showed a significant difference in overall survival （OS） between the two groups. Combining the risk score and clinicopathological characteristics， multivariate Cox regression analysis risk score （HR： 1.136， 95%CI： 1.090~1.183） and age （HR： 1.021， 95%CI： 1.000~1.042） were independent prognostic factors， and a nomogram for PAAD prognostic evaluation was constructed. Furthermore， immunological correlation analysis revealed the differences in immune infiltration and the expression of immune checkpoint genes in the high and low risk groups， and the predictive characteristics of the model were related to immune status. The study constructed a PAAD prognostic model based on 9 aging-related lncRNAs， which may help improve the prognosis management of PAAD patients.

Fermented Epimedium Improves Sperm Deficiency Model in Mice: Effects and Mechanisms
Changze LI, Shuai LIU, Tongxiang DIAO, Keqin ZHANG, Dingqi SUN, Hui ZHANG
2025, 15(1): 170-175. DOI: 10.19586/j.2095-2341.2024.0118
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The incidence of oligospermia and asthenozoospermia in China has been increasing year by year， but there is a lack of effective treatment methods. Therefore， the in-depth development of traditional Chinese medicine has become a research focus. Explored the improvement effect and potential mechanism of precise fermentation of Morinda officinalis by Lactobacillus fermentum Y5 on cyclophosphamide （CTX）?-induced oligoasthenospermia model in mice. Randomly divide the test mice into four groups： control group， model group， unfermented group， and fermented group. Use a sperm analyzer to detect the number and vitality of sperm in the mouse epididymis； Hematoxylin-eosin staining（HE） was performed on mouse testes， and the levels of reactive oxygen species （ROS） and malondialdehyde （MDA） were measured by ELISA. Western blot was used to detect the expression of Keap1/Nrf2/HO-1 signaling pathway proteins in mouse testes. The results showed that compared with the model group， the germ cells in the testes of mice in the fermented and unfermented groups were restored， and the number of sperm in the lumen increased； the sperm count and vitality of mice were significantly improved （P<0.01）； the levels of ROS and MDA in testicular tissue decreased （P<0.01）； Keap1 protein expression decreased， while Nrf2 and HO-1 protein expression increased （P<0.01）； compared with the unfermented group， the improvement effect of the fermented group was more significant （P<0.05）. The results showed that fermented Morinda officinalis can reduce oxidative stress damage and alleviate oligozoospermia in mice through the Keap1/Nrf2/HO-1 signaling pathway， and the improvement effect of precision fermented Morinda officinalis was better than that of unfermented Morinda officinalis.

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