Current Biotechnology

Rational Design Strategies for Advanced Hydrogel-based Sensing Platforms
Yuan SU, Weiwei JIAO, Longjiao ZHU, Wentao XU
2026, 16(2): 215-224. DOI: 10.19586/j.2095-2341.2026.0017
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Sensing technology serves as a cornerstone for health and environmental monitoring， yet its advancement is constrained by the inadequate biocompatibility and mechanical compatibility of traditional interface materials. Hydrogels， as a class of hydrophilic three-dimensional network soft materials， offer an ideal platform to address this bottleneck owing to their biomimetic features and tunable physicochemical properties. For the first time， this review systematically summarized and elaborated on eight rational design paradigms for hydrogel-based sensors， including functional nucleic acid programming， enzymatic catalysis and biomolecular recognition， synthetic supramolecular modules， nanocomposite integration， physical confinement effects， stimulus-responsive polymer systems， microneedle-based sensing， and flexible wearable device integration. These paradigms clarified the central role of hydrogels as either active responsive units or passive optimization matrices across different strategies. The work aims to provide a systematic rational design framework for relevant research， facilitating a paradigm shift in the field from empirical exploration toward rational construction.

Research Progress on Micro-needle Technology and its Mediated Nucleic Acid Delivery in Insects
Shuangshuang YUAN, Sufang ZHANG
2026, 16(2): 225-232. DOI: 10.19586/j.2095-2341.2025.0185
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Micro-needle technology， as an advanced minimally invasive drug delivery platform， has been extensively applied in biomedical research for the transdermal delivery of pharmaceuticals， vaccines， and biological macromolecules. In recent years， modern molecular techniques represented by clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 （CRISPR/Cas9） gene editing and RNA interference （RNAi） have provided revolutionary tools for the research on green prevention and control of agricultural pests. Conventional microinjection methods are cumbersome to operate， low in throughput， and prone to cause damage to insect eggs； while the feeding method is limited by multiple physiological and enzymatic degradation barriers in the insect digestive tract， resulting in difficulty in effective uptake of exogenous dsRNA into the blood circulation， with low and unstable delivery efficiency. In view of this， introducing the mature micro-needle technology （whose core is a micron-scale needle tip array） from the medical field into insect molecular research provides an innovative idea to solve this delivery dilemma. This review systematically explored the application potential of micro-needle technology in insect molecular delivery for the first time， focused on elaborating the types of micro-needles suitable for insect systems， key biocompatible materials and their properties， and deeply analyzed the main opportunities and challenges faced by this interdisciplinary application. The purpose is to provide a new technical perspective and theoretical reference for the development of the next generation of efficient and precise molecular pest control strategies.

Research Progress on the Production Status and Bioactive Substances of Dictyophora
Jize LEI, Gang LIU, Wei JIANG
2026, 16(2): 233-241. DOI: 10.19586/j.2095-2341.2025.0138
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Dictyophora， as a rare edible fungus with a long history and high nutritional value， has been highly respected and favoured by people since ancient times. This paper summarized the research progress on the biological basis， industrial development status， growth requirements and cultivation patterns， nutritional components and bioactive functions of Dictyophora. It was expected to provide theoretical basis and technical support for the sustainable development of the Dictyophora industry and promote the efficient use of Dictyophora resources and industrial upgrading.

Research Progress on Lignocellulose Degraded by Fungi and Their Enzymes
Shan TANG, Siyu LI, Nan DING, Wanyu YU, Hongliang LIU, Xinyue WANG, Shichen HUANG, Minjie FU
2026, 16(2): 242-253. DOI: 10.19586/j.2095-2341.2025.0178
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As the most abundant renewable biomass resource on earth， lignocellulose's efficient conversion is a key approach to addressing energy transition and the "carbon neutrality" goal. However， its inherently complex and dense natural structure poses a severe technical bottleneck due to its recalcitrance to degradation. This review summarized the research progress on the species of fungi involved in lignocellulose degradation and their related enzyme systems. It focused on analyzing the classification characteristics， degradation mechanisms， and extracellular enzyme system compositions of major degrading fungi， such as white-rot fungi， brown-rot fungi， and soft-rot fungi. The catalytic properties and synergistic mechanisms of lignin-degrading enzyme systems （laccase， peroxidase， etc.）， hemicellulases， and cellulases （endoglucanase， exoglucanase， β-glucosidase） were systematically elaborated. The breakthrough role of the newly discovered lytic polysaccharide monooxygenase （LPMO） in improving cellulose degradation efficiency and its synergistic effect with glycoside hydrolases were specifically discussed. Finally， the application prospects and challenges of fungal enzyme systems in fields such as biomass energy and the papermaking industry were prospected， aiming to provide a theoretical reference for the efficient bioconversion of lignocellulosic resources.

Research Progress of Aspergillus niger in Traditional Chinese Medicine, Food and Feed
Ziying YIN, Quan WANG, Zhe WANG, Chen SUN, Zhao WEI, Ziqian YUAN, Yubing FENG
2026, 16(2): 254-259. DOI: 10.19586/j.2095-2341.2025.0062
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Aspergillus niger is an important industrial microorganism with strong enzyme secretion ability and biotransformation characteristics， which has shown extensive potential in the fields of traditional Chinese medicine， food and feed. At present， the industrial application of Aspergillus niger existes multiple bottlenecks such as unclear enzyme production efficiency， fermentation processes requires optimization， and poor cross disciplinary adaptability， meanwhile faces challenges such as technical research and development barriers and limited public awareness. The review summarized the research progress and application status of Aspergillus niger in traditional Chinese medicine， food， and feed， and looked forward to future research directions， in order to provide theoretical basis for further research and application of Aspergillus niger in fermentation related fields.

Molecular Response Mechanism of Microbial Cells to Light-dark Alternation
Jianlei ZHANG, Fanli BU
2026, 16(2): 260-266. DOI: 10.19586/j.2095-2341.2026.0023
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As one of the most fundamental environmental rhythms on earth， light-dark alternation exerts a profound impact on the survival and functions of microorganisms in both natural and artificial environments. Microbes have evolved a series of sophisticated and complex response mechanisms to adapt to such periodic changes in energy and information input. This review systematically summarized the multi-dimensional response mechanisms of microorganisms at the metabolic， transcriptional， and protein levels under light-dark alternation conditions. At the metabolic level， microorganisms dynamically regulate central carbon metabolism， energy metabolism， and redox balance to optimize energy acquisition and substance synthesis. At the transcriptional level， with circadian clock gene networks and light-sensing signal transduction pathways as the core， they globally reprogram gene expression to achieve orderly regulation in the temporal dimension. At the protein level， they rapidly perform adaptive functions through strategies such as post-translational modification， protein stability regulation， and dynamic assembly of protein complexes. This review focused on the interaction and integration of the response mechanisms at these three levels， revealed their inherent temporal and network characteristics， and looked forward to their application prospects in fields such as synthetic biology， biomanufacturing， and environmental remediation.

Research Progress on the Separation, Analysis and Pharmacological Activities of Raffinose Family Oligosaccharides
Pan GUO, Jiaying YANG, Lifeng HAN, Zhifei FU
2026, 16(2): 267-276. DOI: 10.19586/j.2095-2341.2025.0137
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Raffinose family oligosaccharides （RFOs） are a class of functional oligosaccharides widely present in plant tissues， primarily including raffinose， stachyose， and verbascose. In recent years， RFOs have garnered sustained attention from both academia and industry due to their diverse physiological activities and broad application potential. Studies have shown that RFOs exhibit significant prebiotic properties， selectively promoting the proliferation of beneficial bacteria such as Bifidobacterium and Lactobacillus， regulating intestinal microecological balance， and demonstrating potential in modulating lipid metabolism， enhancing immune function， improving glucose metabolism， and protecting the skin barrier. However， challenges remain in the efficient extraction， structural elucidation， and mechanistic understanding of RFOs. This paper systematically reviewed the physicochemical properties， extraction and purification methods， analytical techniques， and research progress of RFOs in fields such as medicine， food， and cosmetics. Future research should focus on the development of green and efficient extraction technologies， in-depth exploration of action mechanisms， and expansion of cross-disciplinary applications to promote the high-value utilization and industrial development of RFOs.

Research Progress on the Active Ingredients and Pharmacological Effects of Fermented Soybean Paste
Honglin CHEN, Jinjin SHEN, Mengge LI, Yan WANG
2026, 16(2): 277-287. DOI: 10.19586/j.2095-2341.2025.0176
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Fermented soybean is a traditional Chinese medicinal herb with a wide range of applications. It was included in the first batch of medicinal and edible substances catalog in 2002 with high research value and application potential. Studies have found that fermented soybean is rich in organic compounds such as volatile substances， flavonoids， plant polysaccharides， amino acids， and inorganic trace elements， exhibiting pharmacological activities including improving Alzheimer's disease， reducing blood sugar and blood lipids， regulating vascular relaxation， anti osteoporosis， anti-tumor， antioxidant， antiviral， and antidepressant. The systematic review summarized the research progress on the active ingredients and pharmacological effects of fermented soybean， in order to provide a reference for the development and comprehensive resource utilization of fermented soybean.

Research Progress on Traditional Chinese Medicine Hippocampus and Strategic Considerations for the Modernization
Wenxuan LIU, Bowen HAN, Wenlong MA, Jiahao ZHAO, Hongshu SUI, Yinqing LI
2026, 16(2): 288-298. DOI: 10.19586/j.2095-2341.2025.0119
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Hippocampus， a valuable traditional marine Chinese medicinal herb in China， possesses therapeutic effects such as warming the kidney and tonifying yang， resolving masses， and reducing swelling. Studies have shown that Hippocampus contains various bioactive components including amino acids， peptides， steroids， fatty acids， and trace elements， and exhibits pharmacological activities such as sex hormone-like effects， antioxidant， anti-tumor， and anti-fatigue properties， thereby having wide applications in clinical treatment and healthcare fields. Due to overfishing and ecological destruction， wild Hippocampus resources have been depleted. All species are listed in the National Key Protected Wildlife List and Appendix Ⅱ of Convention on International Trade in Endangered Species of Wild Fauna and Flora （CITES）， with their commercial trade strictly restricted. Artificial cultivation has become the key to addressing the resource dilemma； however， most maturely cultivated species have not yet been included in the Chinese Pharmacopoeia. The Pharmacopoeia lacks sufficient standards for species identification and quality control of Hippocampus， leading to chaotic market origins and circulation of counterfeit products， which restricts the legitimate utilization of cultivated varieties. This review systematically summarized the current status of Hippocampus resources， their chemical components， pharmacological effects， quality standards， and progress in product transformation， analyzed the potential and bottlenecks of non-Pharmacopoeia cultivated varieties， and proposed targeted development strategies to promote the sustainable utilization of Hippocampus resources and the modernization of traditional Chinese medicine.

Research Progress on Effects of Medicinal and Food Homologous Substances on Skin Health
Ge YAN, Kunlun HUANG, Tao TONG
2026, 16(2): 299-308. DOI: 10.19586/j.2095-2341.2025.0191
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With the intensification of population aging， medicinal and food homologous substances， which possess both dietary and medicinal values， have garnered increasing attention in the field of skin health. Current studies have reported the positive effects of various medicinal and food homologous substances in alleviating skin inflammation， promoting wound healing， removing spots and whitening， delaying skin photoaging and natural aging， as well as improving acne. This article systematically reviewed the current research status of medicinal and food homologous substances in improving skin health， and analyzed the active components， efficacy， and molecular mechanisms of representative substances such as licorice， astragalus， poria， and kudzu root. Furthermore， it discussed and prospected future research directions in terms of mechanistic insights， clinical translation potential， and systematic efficacy evaluation. The aim of this paper is to provide a theoretical reference for further research and product development of medicinal and food homologous substances in the field of skin health.

Cytochrome P450 Enzyme System in Drug Development: Advances in Research on Regulation of Hepatic Drug Metabolism and Clinical Applications
Yumeng WANG, Pengxiang ZHAO, Xujuan ZHANG, Zheng DANG, Han LI, Ziyi LIU, Jiateng WANG, Sixue YU, Xuemei MA
2026, 16(2): 309-317. DOI: 10.19586/j.2095-2341.2025.0126
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The liver， as the central organ of human metabolism and detoxification， maintains its functional integrity through a complex network of enzyme systems. Within this network， the cytochrome P450 （CYP450） enzyme system assumes a pivotal role by catalyzing phase Ⅰ drug metabolism， thereby mediating the detoxification or toxic biotransformation of exogenous compounds. Modulation of CYP450 activity exerts a profound impact on drug efficacy and drug safety. This enzyme system also plays critical therapeutic and pathophysiological roles in the development and progression of diverse diseases， including liver disorders， cardiovascular diseases， and cancer. In-depth investigations into CYP450 functions and their disease-associated alterations provide a fundamental basis for developing novel therapeutic strategies. The article centered on elucidating the clinical applications of CYP450 inhibitors and inducers， while summarizing the critical significance of targeted CYP450 regulation in optimizing drug metabolism， enhancing therapeutic efficacy， reducing adverse reactions， and promoting personalized medicine， aiming to offer new avenues for precision treatment of liver diseases and expand their applications.

Mitochondrial Alterations in Hepatocellular Carcinoma and Advances in Targeted Therapeutics
Xueqi HAN, Peixian GAO, Xujuan ZHANG, Pengxiang ZHAO, Mengyu LIU
2026, 16(2): 318-326. DOI: 10.19586/j.2095-2341.2025.0183
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Hepatocellular carcinoma （HCC） is a malignancy associated with high global incidence and mortality rates， whose development and progression are closely linked to metabolic dysregulation. As central hubs for cellular energy metabolism and signaling， mitochondria undergo extensive and complex remodeling in HCC， profoundly influencing tumor initiation， progression， and therapeutic response. However， the specific mechanisms underlying these mitochondrial changes and their exact roles in HCC pathogenesis remain incompletely elucidated， which has， to a certain extent， impeded the development and clinical application of mitochondria-targeted therapeutics for HCC. This review systematically summarized mitochondrial alterations in HCC from three key dimensions： metabolic reprogramming， oxidative stress and mitochondrial DNA （mtDNA） damage， and dysregulation of mitochondrial dynamics. It further elucidated the mechanisms by which these alterations drive HCC development and treatment response. Additionally， the review consolidated and prospected the current progress on mitochondria-targeted therapeutic strategies for HCC， aiming provide theoretical foundation and novel insights for future basic research and clinical interventions in the treatment of HCC.

Advances on Mechanisms of Pyroptosis and Ferroptosis in End-stage Renal Disease and Nutritional Interventions
Qian YANG, Wei QIN
2026, 16(2): 327-337. DOI: 10.19586/j.2095-2341.2025.0189
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Chronic end-stage renal disease （ESRD） represents a major challenge in modern nephrology. Its progression is closely associated with programmed cell death （PCD）， among which pyroptosis and ferroptosis play core regulatory roles. These PCD processes are regulated not only by genetic and pathological signals but also significantly dependent on nutritional status and metabolic energy supply. Although studies on the “nutrition-PCD-ESRD” axis have been increasing， relevant evidence remains fragmented and mechanistic understanding has not been unified. Accordingly， this review systematically summarized the molecular mechanisms of pyroptosis and ferroptosis in patients with ESRD and maintenance hemodialysis （MHD）. Pyroptosis contributes to irreversible kidney injury through a three-stage pathway： initiation， amplification， and propagation. Ferroptosis is driven by the core axis of iron overload， antioxidant imbalance， and lipid peroxidation amplification. Both pathways are key drivers of irreversible renal damage. This review further focused on the intervention effects of nutrients including zinc， vitamin D， ω-3 polyunsaturated fatty acids， selenium， vitamin E， L-carnitine， and dietary fiber by targeting key nodes of the above PCD pathways. It confirmed that nutritional interventions can delay disease progression through multiple pathways， such as inhibiting inflammasome activation， restoring antioxidant defense， and regulating iron metabolism. Meanwhile， current challenges in precision nutritional therapy were identified， including high patient heterogeneity， insufficient evidence strength， and limitations of single interventions. Future research directions were proposed， such as establishing a PCD-signature-based precise stratification system， developing multi-target synergistic intervention strategies， and strengthening supporting technologies. This review provided mechanistic evidence and translational ideas for nutritional intervention in ESRD/MHD patients， and offered a reference for basic research and clinical strategies.

Research Progress on Reciprocal Interactions Between Obstructive Sleep Apnea and Gut Microbiota
Jing QI, Xiuyun LYU
2026, 16(2): 338-347. DOI: 10.19586/j.2095-2341.2025.0162
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Obstructive sleep apnea （OSA） is a common chronic sleep-related breathing disorder， characterized primarily by intermittent hypoxia and sleep fragmentation resulting from the recurrent collapse of the upper airway during sleep. Studies indicated that disturbances of the gut microbiota represent a crucial intermediary mechanism in the multi-system pathological damage and development of comorbidities induced by OSA. Changes in the gut microbiota under OSA conditions， including abnormalities in α/β diversity， enrichment of anaerobic bacteria， reduction in short-chain fatty acid-producing bacteria， as well as alterations in metabolic profiles such as bile acids， amino acids， and fatty acids， were systematically elucidated. Furthermore， the key driving role of gut microbiota and their metabolites in OSA-related comorbidities， such as hypertension and atherosclerosis， was extensively investigated. Concurrently， research progress on intervention strategies targeting the gut microbiome was comprehensively integrated. These findings provide new perspectives and a foundation for mechanistic research and clinical management of OSA.

Research Progress on the Prevention and Treatment of Cardiotoxicity Caused by Chemotherapy Drugs
Yan CHEN, Zhida LIU
2026, 16(2): 348-355. DOI: 10.19586/j.2095-2341.2026.0008
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With the continuous innovation and development of anti-tumor treatment methods， the prognosis of cancer patients has been significantly improved. However， the problem of cardiotoxicity induced by chemotherapeutic drugs has become increasingly prominent. This toxic reaction not only severely limits the choice of treatment regimens for cancer patients but also exerts an important impact on their quality of life and long-term prognosis. This article systematically summarized the clinical manifestations and action mechanisms of cardiotoxicity associated with common chemotherapeutic drugs， reviewed the research progress in prevention and treatment at home and abroad， and sorted out the existing common and new prevention and treatment strategies， in order to provide theoretical reference and practical basis for the clinical management of chemotherapy-induced cardiotoxicity.

GeoCas9-mediated sgRNA-free Short Double-stranded DNA Rapid Single-stranded Method and its Performance Characterization
Xinran LI, Chenyi WANG, Xin QI, Yaning GUO, Fukai LI, Jian ZHOU, Mengrui YANG, Liang LI, Kai LI
2026, 16(2): 356-364. DOI: 10.19586/j.2095-2341.2026.0028
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Efficient conversion of short double?stranded DNA （dsDNA） into single?stranded DNA is a critical pretreatment step in molecular diagnostics， genome editing， and DNA nanostructure construction. However， existing methods mainly rely on thermal or chemical denaturation strategies， which suffer from limitations such as complex operation， poor reversibility， and potential damage to DNA integrity. Herein， we reported a novel rapid in vitro strategy for the single?stranding of short dsDNA based on the CRISPR?associated protein GeoCas9. Results showed that GeoCas9 can bind and unwind short dsDNA substrates with lengths ranging from 20 to 60 bp in the absence of sgRNA guidance， enabling a non?cleaving， reversible， and highly efficient single?stranding process under ambient conditions. We systematically evaluated the effects of protein concentration， protein?to?substrate molar ratio， reaction time， and dsDNA length on the single?stranding efficiency， and determined the optimal reaction parameters： a final GeoCas9 concentration of 50 nmol·L^-1 and a protein?to?dsDNA molar ratio of 1∶1 （dsDNA concentration 50 nmol·L^-1）. Under optimized conditions， this method achieves rapid and stable duplex unwinding in a mild environment while maintaining DNA structural integrity. This study expands the functional understanding of Cas9 proteins and provides a simple， efficient， and well?controllable technical route for precise nucleic acid manipulation and downstream bioanalysis.

Analysis of Gene Diversity in GH10 and GH11 Families in the Rumen of Domestic Sheep from the Pamir Plateau in Xinjiang
Zhulin TAO, Yutao WANG
2026, 16(2): 365-371. DOI: 10.19586/j.2095-2341.2025.0097
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By analyzing the diversity of GH10 and GH11 genes in the rumen of domestic sheep from the Pamir Plateau in Xinjiang， this study provides new genetic resources for elucidating the enzymatic properties of GH10 and GH11 family xylanases， while also laying a foundation for understanding the structure of rumen xylanases. This study collected rumen contents from six domestic sheep on the Pamir Plateau in Xinjiang. Total genomic DNA was extracted， and universal primers specific to each family were used for amplification. High-throughput sequencing was performed， and bioinformatics methods were applied to analyze the sequencing results. Following quality control， operational taxonomic unit （OTU） clustering analysis identified that the GH10 gene family contained a total of 2 040 operational taxonomic units （OTUs）， with 18 OTUs showing relative abundance greater than 0.01； the GH11 gene family contained a total of 825 OTUs， with 14 OTUs having a relative abundance greater than 0.01. NCBI homology comparisons revealed that GH10 sequences were classified into 4 phyla， 5 families and 6 genera， with 97.79% of counts remaining unclassified， while GH11 sequences were assigned to 4 phyla， 8 families and 10 genera， with 69.94% of counts unclassified. Alpha diversity analysis demonstrated that GH10 exhibited higher diversity， evenness and richness indices compared to GH11. The rumen of domestic sheep in the Xinjiang Pamir Plateau harbors abundant GH10 and GH11 family xylanases， which exhibit certain specificity and the GH10 gene family displays higher diversity than the GH11 gene family.

Investigation on the Alleviative Effects of Hylocereus undatus on Intestinal Damage in Drosophila melanogaster
Ju HAN, Jiapeng CHEN, Qi ZHOU, Hongrui CHEN, Dachang WU
2026, 16(2): 372-379. DOI: 10.19586/j.2095-2341.2025.0144
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The study aims to investigate the alleviating effect of Hylocereus undatus on the intestinal damage in fruit flies， and explore the possible mechanism， with the goal of providing theoretical basis for the development and application of H. undatus. An acute intestinal injury model of wild-type Drosophila melanogaster was induced by dextran sulfate sodium salt （DSS）. The effects of feeding normal medium and medium containing different concentrations （0.5%， 1%， 5%） of H. undatus on the survival indicators （locomotor ability， activity and survival rate）， antioxidant indicators （superoxide dismutase， catalase， reduced glutathione， malondialdehyde）， and intestinal injury conditions （smurf assay， lactate dehydrogenase， Ca²⁺ content determination） of Drosophila melanogaster were analyzed. The results showed that the acute intestinal injury model of Drosophila melanogaster was successfully constructed by DSS. After feeding with medium containing H. undatus， the survival rate， locomotor ability and activity of Drosophila melanogaster were significantly improved， the activities of superoxide dismutase， catalase and lactate dehydrogenase and the content of reduced glutathione increased， while the contents of Ca²⁺ and malondialdehyde decreased， and the intestinal injury was significantly alleviated. Moreover， the high concentration group （containing 5% H. undatus） had the most significant effect. H. undatus has an alleviating effect on the intestinal injury induced by DSS in Drosophila melanogaster， which may be related to the repair of intestinal cell damage and the enhancement of antioxidant capacity by H. undatus.

Heterologous Expression and Characterization of a Novel Chitinase Derived from Talaromyces flavus
Yang WANG, Songni YU, Yahua LI, Hongquan XIAN
2026, 16(2): 380-387. DOI: 10.19586/j.2095-2341.2025.0100
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Talaromyces flavus is a mycoparasitic fungus， and the chitinases it producs play an important role in the mycoparasitic process， showing high application value in biological control of plant diseases and chitin degradation. In this study， intronless chitinase genes were cloned from T. flavus using PCR technology， expressed in the Pichia pastoris system， and the characteristics of chitinases were analyzed. The results showed that the ORF sizes of chitinase genes 04446 and 06441 were 1 032 and 1 029 bp， respectively. The P. pastoris expression vectors pPIC9K/04446 and pPIC9K/06441 were successfully constructed， and highly expressed yeast engineering strains GS115/04446-45 and GS115/06441-57 were obtained； the purified protein Tfch04446 expressed by 04446 had a molecular weight of 36.8 kD， and the protein Tfch06441 expressed by 06441 had a molecular weight of 37.2 kD. The optimal reaction temperature of chitinase Tfch04446 was 45 ℃， while that of chitinase Tfch06441 was 50 ℃. The optimal reaction pH was 6 for both Tfch04446 and Tfch06441. For Tfch04446， the K_m value was 2.27 mg·mL^-1 and the maximum reaction rate （V_max） was 0.085 mg·mL^-1·min^-1； for Tfch06441， the K_m value was 1.52 mg·mL^-1 and the V_max was 0.067 mg·mL^-1·min^-1. Regarding metal ions， Ca²⁺ and Ba²⁺ exerted a promoting effect on the activity of chitinase Tfch04446， while NH₄⁺ showed a certain degree of inhibitory effect. For chitinase Tfch06441， Ba²⁺ had a promoting effect on its activity， whereas Na⁺， K⁺， Ca²⁺ and NH₄⁺ all exhibited inhibitory effects on it. The research results laid a foundation for the development and utilization of T. flavus and its chitinases.

Transfer of the pSLT Plasmid and its Impact on Intestinal Cell Apoptosis
Tian HONG, Lulu ZHAO, Yifan DU, Jingwen LI, Erning CHEN, Meihong DU
2026, 16(2): 388-394. DOI: 10.19586/j.2095-2341.2025.0090
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The pSLT plasmid is a specific virulence plasmid primarily found in Salmonella enterica serovar Typhimurium strains. This study aimed to explore the feasibility of horizontal gene transfer （HGT） of the Salmonella pSLT virulence plasmid and investigate its effects on cell necrosis and apoptosis. The Kanamycin resistance gene derived from plasmid pKD46 was integrated into the Salmonella pSLT plasmid using λRed homologous recombination technology. Salmonella enterica serovar Typhimurium （ATCC 14028） served as the donor strain， and Escherichia coli （ATCC 25922） as the recipient strain. Conjugation was performed using the M9 minimal medium plate mating method. The obtained transconjugants were cultured in Kanamycin-supplemented MacConkey broth to verify acid and gas production. Single colonies were isolated， followed by confirmatory PCR and sequencing for the inserted Kana gene. Human colon carcinoma cells （HCT116）， representing intestinal cells， were infected with transconjugants at a multiplicity of infection （MOI） of 100. Cell necrosis and apoptosis were assessed using flow cytometry （Annexin V/PI staining）. The results showed that single colonies grew on nutrient agar plates containing Kanamycin， confirming successful recombination and insertion of the Kana gene fragment into pSLT （creating pSLT^Kana）. Transconjugants exhibited acid and gas production in Kanamycin-supplemented MacConkey broth， demonstrating the successful conjugative transfer of the pSLT^Kana plasmid to Escherichia coli. Compared to the control group， the ATCC 25922 （wild-type E. coli） treatment group did not cause a significant change （P>0.05） in the proportion of Annexin V/PI double-positive cells （indicating late apoptosis/necrosis）. In contrast， both the ATCC 25922 pSLT^Kana （E. coli harboring pSLT） and ATCC 14028 （Salmonella with native pSLT） treatment groups showed a significant reduction （P<0.01） in the proportion of Annexin V/PI double-positive cells. By constructing the ATCC 14028 pSLT^Kana recombinant strain， this study confirmed the feasibility of HGT of the pSLT virulence plasmid from Salmonella to E. coli. The ATCC 25922 pSLT^Kana strain exhibited a potential inhibitory effect on necrosis and apoptosis in intestinal cells. These findings provide crucial scientific data and insights for clinical treatment strategies and public health safety oversight.

Mechanism of SIRT1 in Regulating Learning and Memory via Modulating Neurotransmitter Metabolic Homeostasis
Yanan QIAO, Yubing ZENG, Xuejia HE, Fan LIU, Shan WANG
2026, 16(2): 395-411. DOI: 10.19586/j.2095-2341.2025.0154
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SIRT1， an NAD⁺-dependent deacetylase， plays a pivotal role in maintaining neuronal function and metabolic homeostasis. However， its molecular mechanisms in regulating neurotransmitter metabolism and cognitive impairment remain unclear. In this study， we established a Sirt1-deficient mouse model （Sirt1^+/- ） and performed transcriptomic and metabolomic analyses on hippocampus to systematically investigate the impact of SIRT1 loss on cerebral neurotransmitter metabolic networks and cognitive function. Phenotypic observations revealed that Sirt1^+/- mice exhibited significant deficits in spatial memory and learning abilities. Metabolomic and transcriptomic profiling demonstrated that SIRT1 deficiency led to metabolic dysregulation of key neurotransmitters， including glutamate and γ-aminobutyric acid （GABA） in the hippocampus， accompanied by marked upregulation of neurotransmitter metabolism-related genes like Gad1 and Gad2. Integrated multi-omics analysis further confirmed that SIRT1 deficiency suppressed neuroactive ligand-receptor interactions and cAMP signaling pathways， resulting in disrupted neurotransmitter homeostasis and subsequent cognitive dysfunction. In conclusion， this study elucidates a novel mechanism by which SIRT1 maintains cognitive function through coordinated reprogramming of neurotransmitter metabolism and energy homeostasis， providing potential therapeutic targets for neurodegenerative disorders.

ALKBH5 Affects Cisplatin Chemosensitivity in Non-small Cell Lung Cancer by Regulating CPNE1 m⁶A Methylation
Hongfei LIU, Wen ZHAO, Huiying YANG
2026, 16(2): 412-421. DOI: 10.19586/j.2095-2341.2025.0164
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This study aimed to investigate the effect and mechanism of the N6-methyladenosine （m⁶A） demethylase alkB homolog 5 （ALKBH5） on cisplatin （DDP） chemosensitivity in non-small cell lung cancer （NSCLC） by regulating the m⁶A methylation of CPNE1 （Copine 1） mRNA. The expression levels of ALKBH5 and CPNE1 in lung cancer cell lines were detected by qRT-PCR and Western blot. The sequence-based RNA adenosine methylation site predictor （SRAMP） was used to predict the m⁶A methylation sites of CPNE1， Me-RIP assay was performed to detect its methylation level， and RNA immunoprecipitation （RIP） assay was conducted to verify the binding between ALKBH5 and CPNE1 mRNA. Lung cancer H157 cells were transfected with OE-CPNE1， si-CPNE1， and si-ALKBH5， respectively. Cell viability was detected by CCK-8 assay to calculate the IC₅₀ value， colony formation assay was used to evaluate cell proliferation， and terminal deoxynucleotidyl transferase dUTP nick end labeling （TUNEL） staining was employed to detect cell apoptosis. The PI3K/AKT activator 740Y-P was added to si-CPNE1-transfected cells to analyze the role of CPNE1 in the PI3K/AKT pathway. The results showed that the expressions of ALKBH5 and CPNE1 in NSCLC cells were significantly higher than those in normal lung epithelial cells （P<0.05）. Knockdown of ALKBH5 decreased the expression of CPNE1 and increased its m⁶A methylation level （P<0.05）. The RIP assay confirmed a direct binding between ALKBH5 and CPNE1 mRNA. Inhibition of ALKBH5 or CPNE1 significantly enhanced the cytotoxic effect of cisplatin on lung cancer cells， which was manifested by decreased cell viability， inhibited proliferation， and increased apoptosis （P<0.05）. In addition， the PI3K/AKT activator 740Y-P could rescue the pathway inhibition and chemoresistance phenotype induced by CPNE1 knockdown. These findings indicated that ALKBH5 upregulated CPNE1 expression in an m⁶A-dependent manner and activated the PI3K/AKT pathway， thereby reducing the cisplatin chemosensitivity of NSCLC cells.

ZG16 Inhibits EMT in Hepatocellular Carcinoma via CTNNA3 and Negatively Regulates of PD-L1
Ce JIANG, Yi LIU, Yalei ZHAO, Haiyun ZHANG, Xiangrong CHANG
2026, 16(2): 422-431. DOI: 10.19586/j.2095-2341.2025.0152
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To investigate the effect of zymogen granule protein 16 （ZG16） on epithelial mesenchymal transition （EMT） in hepatocellular carcinoma cells and its potential association with the immune checkpoint molecule programmed death-ligand 1 （PD-L1）. The expression differences of ZG16 between hepatocellular carcinoma （HCC） and normal liver tissues， as well as its relationship with patient prognosis， were analyzed based on the GEO database and the KM-Plot website. Huh7 and HepG2 cells were transfected with the pcDNATM3.1-ZG16 overexpression plasmid to overexpress ZG16. Protein expression levels of ZG16， CTNNA3， E-cadherin， N-cadherin， Vimentin and PD-L1 were detected by Western blot. The localization and protein expression levels of ZG16 and CTNNA3 were examined using immunofluorescence. The interaction between ZG16 and CTNNA3 was detected by co-immunoprecipitation. Cell migration ability was assessed through scratch wound healing assays， and cell colony formation ability was evaluated using colony formation assays.Bioinformatic analysis revealed that ZG16 expression was significantly downregulated in HCC tissues， and higher ZG16 expression was associated with significantly improved patient survival （P<0.05）. In vitro experiments showed that compared to the control group， ZG16 overexpression significantly increased E-cadherin expression and decreased N-cadherin and Vimentin expression （P<0.05）， along with significantly reduced cell migration and colony formation abilities in Huh7 and HepG2 cells （P<0.05）. In HCC patients， ZG16 expression levels were negatively correlated with PD-L1， and ZG16 overexpression significantly reduced PD-L1 protein levels. Mechanistically， ZG16 overexpression significantly increased CTNNA3 protein levels （P<0.05）. Co-immunoprecipitation and co-localization immunofluorescence confirmed that ZG16 and CTNNA3 interact and co-localize in the cytoplasm. Functional rescue experiments demonstrated that， compared to Huh7 cells overexpressing ZG16 alone， simultaneous overexpression of ZG16 and knockdown of CTNNA3 promoted the epithelial-mesenchymal transition process and enhanced cell migration and colony formation abilities （P<0.05）. ZG16 upregulates CTNNA3 to inhibit the EMT process in hepatocellular carcinoma cells. Additionally， ZG16 negatively regulates PD-L1 protein expression. The research results can provide new molecular targets and theoretical basis for the clinical treatment of liver cancer.

Evaluation of the Preventive and Reparative Effects of N-PS on Skin Barrier Impairment
Xian CHENG, Zhulan ZHANG, Zhouqin CHEN, Xiaojing GAO, Huangjian YANG, Lingbin YAN, Yunyang LIAN
2026, 16(2): 432-438. DOI: 10.19586/j.2095-2341.2025.0160
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This study aimed to investigate the preventive and reparative effects of Nocardia rubra cell wall skeleton polysaccharide components （N-PS） on skin barrier impairment. The antioxidant activity of N-PS was evaluated， together with its effects on phagocytic activity of RAW 264.7 cells， melanin synthesis and tyrosinase activity in B16 mouse melanoma cells， as well as the reparative effect on HaCaT cells injured by H₂O₂. Within the concentration range of 0.25~4.00 mg·mL^-1， N-PS showed strong DPPH free radical scavenging activity， with a maximum scavenging rate of 40.0%. Stimulation with 200 μg·mL^-1 N-PS increased the proliferation rate of RAW 264.7 cells by 2.56%， enhanced the phagocytic activity by 26.94%， and elevated the macrophage-mediated innate immune response by 65.88%. Meanwhile， N-PS reduced the proliferation of B16 mouse melanoma cells by 36.89%， inhibited melanin synthesis by 39.05%， and decreased total cellular tyrosinase activity by 38.86%. N-PS exhibited significant protective effects on H₂O₂-damaged HaCaT cells， and the cell viability increased from 65.62% in the blank control group to 97.79% after pretreatment with 800 μg·mL^-1 N-PS. In summary， N-PS showed a maximum DPPH scavenging activity of 40.0% at 0.25~4.00 mg·mL^-1， and 200 μg·mL^-1 N-PS exerted favorable preventive and reparative effects on skin barrier impairment.

Based on Network Pharmacology and Mendelian Randomization to Investigate the Mechanism of Hyssopus cuspidatus Boriss. in the Treatment of Asthma
Mingchi LI, Zengfang SHI, Guoshuai ZHANG, Qi YAN, Jiale WANG, Xinke ZHANG, Linfang HUANG
2026, 16(2): 439-451. DOI: 10.19586/j.2095-2341.2025.0155
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This study aims to systematically investigate the active components， target genes， and molecular mechanisms of Hyssopus cuspidatus Boriss. in the treatment of asthma， thereby providing a scientific basis for its clinical application. A comprehensive metabolomic analysis of Hyssopus cuspidatus Boriss. was conducted using dual-platform UPLC-MS/MS and GC-MS to identify its active components. Potential therapeutic targets were screened using databases such as TCMSP， PubChem， and SwissTargetPrediction. Asthma-related targets were retrieved from GeneCards， OMIM， and TTD databases. A protein-protein interaction （PPI） network was constructed， followed by Gene Ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） enrichment analyses. Molecular docking was performed to validate the binding affinity between active components and core targets. Finally， a two-sample Mendelian randomization （MR） analysis was employed to verify the causal relationship between core targets and asthma. The results showed that a total of 79 active components and 410 potential targets were identified in Hyssopus cuspidatus Boriss. Intersection with asthma-related targets yielded 76 common targets. PPI network analysis revealed AKT1， STAT3， EGFR， PTGS2， and SRC as core targets. GO and KEGG enrichment analyses indicated significant enrichment in processes such as the PI3K/AKT signaling pathway， inflammatory response， and apoptosis. Molecular docking results demonstrated strong binding affinities between flavonoid components （e.g.， luteolin and isorhamnetin） and core targets. Mendelian randomization analysis further confirmed a significant causal relationship between AKT1 and asthma. Hyssopus cuspidatus Boriss. might exert therapeutic effects on asthma through multiple flavonoid active components targeting core genes such as AKT1， regulating signaling pathways like PI3K/AKT， thereby suppressing airway inflammation and alleviating airway hyperresponsiveness. AKT1 is identified as a key target with potential clinical translational value.

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