Hydrogenases, as a class of highly efficient and reversible biological catalysts, can catalyze the reduction of protons to molecular hydrogen, thus demonstrating great potential in a wide range of fields such as renewable energy production and green chemistry. Despite their significant potential, the large-scale industrial application of hydrogenases has long been constrained by several inherent limitations, including high sensitivity to molecular oxygen, the challenges in the in vitro reconstitution and maturation of their catalytic centers, and the inefficiency and instability of the natural electron transfer pathways. To overcome these limitations and enhance the catalytic performance of hydrogenases, researchers have developed various strategies, among which enzyme molecular engineering, photo-driven modification, and enzyme immobilization techniques are the most common exploration directions. Particularly, enzyme immobilization technology is widely used to improve the reusability of hydrogenases, but traditional immobilization methods often come with disadvantages in practical applications, such as complex multi-step procedures and insufficient biocompatibility of the immobilization materials. In recent years, bioencapsulation technology has emerged as a promising alternative strategy to enhance the catalytic performance of hydrogenases. This method utilizes biologically derived encapsulation materials to construct physically confined and precisely defined chemical microenvironments around the enzyme molecules, offering simpler self-assembly processes and superior biocompatibility. With these biomimetic constructs, bioencapsulation technology not only provides better oxygen tolerance but also helps to create a local microenvironment conducive to sustained catalytic function. This article systematically reviews the latest research progress of two main bioencapsulation strategies for hydrogenases: one is the encapsulation technology based on protein-based nanocages; the other is the engineering strategy for whole-cell hydrogenase expression. In the nanocage-based systems, this article focuses on the structural and functional characteristics of virus-like capsids and carboxysome protein shells, which serve as efficient enzyme encapsulation scaffolds, not only providing a stable physical barrier to prevent oxygen diffusion but also enabling high-density enzyme loading, thereby promoting substrate channeling effects and electron transfer kinetics. This article also discusses whole-cell encapsulation systems, which achieve hydrogenase compartmentalization within engineered cellular structures or by using external natural polysaccharide-based encapsulation matrices to wrap whole-cell catalysts. Bioencapsulation strategies can bring multiple synergistic benefits: they can effectively protect hydrogenases from oxygen-mediated inactivation, significantly delay the decline of catalytic activity over time, and enhance the hydrogen production rate by increasing the local concentration of active enzyme molecules and optimizing the electron transfer efficiency from redox partners to the catalytic center.Despite the significant progress made, several technical challenges remain to be addressed. The main obstacles include limited enzyme loading and encapsulation efficiency, insufficient long-term stability of encapsulation materials under operating conditions, and the need to improve the matching of the photo-biological interface in systems integrating light-harvesting components with enzymatic catalysis. Future efforts can focus on the integration of multiple technological approaches, such as using computer-aided protein design to optimize encapsulation structures, developing engineered electron transfer pathways to enhance catalytic conversion efficiency, and designing composite multifunctional materials with both structural stability and functional adaptability. These directions collectively aim to achieve efficient, stable, and scalable hydrogen production applications of bioencapsulated hydrogenase systems.

Hydrogenases, as a class of highly efficient and reversible biological catalysts, can catalyze the reduction of protons to molecular hydrogen, thus demonstrating great potential in a wide range of fields such as renewable energy production and green chemistry. Despite their significant potential, the large-scale industrial application of hydrogenases has long been constrained by several inherent limitations, including high sensitivity to molecular oxygen, the challenges in the in vitro reconstitution and maturation of their catalytic centers, and the inefficiency and instability of the natural electron transfer pathways. To overcome these limitations and enhance the catalytic performance of hydrogenases, researchers have developed various strategies, among which enzyme molecular engineering, photo-driven modification, and enzyme immobilization techniques are the most common exploration directions. Particularly, enzyme immobilization technology is widely used to improve the reusability of hydrogenases, but traditional immobilization methods often come with disadvantages in practical applications, such as complex multi-step procedures and insufficient biocompatibility of the immobilization materials. In recent years, bioencapsulation technology has emerged as a promising alternative strategy to enhance the catalytic performance of hydrogenases. This method utilizes biologically derived encapsulation materials to construct physically confined and precisely defined chemical microenvironments around the enzyme molecules, offering simpler self-assembly processes and superior biocompatibility. With these biomimetic constructs, bioencapsulation technology not only provides better oxygen tolerance but also helps to create a local microenvironment conducive to sustained catalytic function. This article systematically reviews the latest research progress of two main bioencapsulation strategies for hydrogenases: one is the encapsulation technology based on protein-based nanocages; the other is the engineering strategy for whole-cell hydrogenase expression. In the nanocage-based systems, this article focuses on the structural and functional characteristics of virus-like capsids and carboxysome protein shells, which serve as efficient enzyme encapsulation scaffolds, not only providing a stable physical barrier to prevent oxygen diffusion but also enabling high-density enzyme loading, thereby promoting substrate channeling effects and electron transfer kinetics. This article also discusses whole-cell encapsulation systems, which achieve hydrogenase compartmentalization within engineered cellular structures or by using external natural polysaccharide-based encapsulation matrices to wrap whole-cell catalysts. Bioencapsulation strategies can bring multiple synergistic benefits: they can effectively protect hydrogenases from oxygen-mediated inactivation, significantly delay the decline of catalytic activity over time, and enhance the hydrogen production rate by increasing the local concentration of active enzyme molecules and optimizing the electron transfer efficiency from redox partners to the catalytic center.Despite the significant progress made, several technical challenges remain to be addressed. The main obstacles include limited enzyme loading and encapsulation efficiency, insufficient long-term stability of encapsulation materials under operating conditions, and the need to improve the matching of the photo-biological interface in systems integrating light-harvesting components with enzymatic catalysis. Future efforts can focus on the integration of multiple technological approaches, such as using computer-aided protein design to optimize encapsulation structures, developing engineered electron transfer pathways to enhance catalytic conversion efficiency, and designing composite multifunctional materials with both structural stability and functional adaptability. These directions collectively aim to achieve efficient, stable, and scalable hydrogen production applications of bioencapsulated hydrogenase systems.

Exercise, as a non-pharmacological intervention, holds a pivotal role in metabolic regulation, neuroplasticity, and immune homeostasis maintenance. However, human exercise studies are constrained by ethical limitations in tissue sampling, especially for key organs such as muscles and the brain. Meanwhile, rodent models like mice exhibit physiological differences in exercise patterns and metabolic rates from human. Despite these challenges, approximately 70% of human and mouse genes are conserved, providing a molecular basis for cross-species comparisons. This paper leverages the GEPREP database, which integrates human and mouse exercise transcriptomic data from multiple platforms, to conduct a comprehensive cross-species analysis of exercise-induced gene expression patterns. We employ a stringent data standardization process, including the conversion of orthologous genes and the filtering of low-expressing genes, to ensure the accuracy and reliability of the analysis. A mixed-effects model is utilized to assess differential gene expression across multiple cohorts, identifying genes that are significantly upregulated or downregulated in response to exercise. The analysis reveals a complex pattern of gene expression, with a significant number of genes showing conserved responses between humans and mice, particularly in acute aerobic exercise, where genes such as ATF3, PPARGC1A, and ANKRD1 are commonly upregulated. These genes are implicated in muscle stress response, metabolic regulation, and muscle adaptation, highlighting the shared molecular pathways activated by exercise across species. However, the study also uncovers substantial species-specific differences in gene expression, especially in chronic aerobic exercise, where the number of divergently regulated genes increases. These differences suggest that while some fundamental biological processes are conserved, the specific regulatory mechanisms and gene expression patterns can vary significantly between humans and mice. Functional enrichment analysis further reveals that conserved genes are involved in muscle development, inflammation regulation, and energy metabolism, while species-specific genes are associated with ion transport, extracellular matrix (ECM) organization, and muscle contraction, indicating the multifaceted impact of exercise on skeletal muscle function. The findings emphasize the importance of considering species-specific differences when interpreting results from animal models and translating them to human health applications. The study highlights the need for a more nuanced understanding of the molecular underpinnings of exercise-induced adaptations and underscores the value of cross-species comparative analyses in uncovering the evolutionary and functional basis of these responses. Future research should focus on integrating multi-omics data and expanding the analysis to include other tissues to provide a more comprehensive view of the systemic effects of exercise. Additionally, the development of species-specific gene editing models and the validation of key genes in exercise physiology will further enhance our understanding of the evolutionary logic behind exercise interventions. This study not only provides valuable insights into the molecular mechanisms of exercise-induced adaptations but also underscores the necessity of validating findings from animal models in human cohorts to ensure the reliability and applicability of translational research in exercise science. By addressing these aspects, the study aims to bridge the gap between basic research and clinical applications, ultimately contributing to the development of personalized exercise prescriptions and interventions that can effectively promote health and prevent diseases.

BACKGROUND:Metformin is currently considered the first-line medication for the treatment of type 2 diabetes.Metformin may delay the progression of osteoarthritis,but its specific mechanism of action remains unclear.OBJECTIVE:To evaluate the therapeutic effects and the related action mechanisms of metformin on osteoarthritis in rats.METHODS:(1)Network pharmacology:Potential common targets for metformin,osteoarthritis,and ferroptosis were screened using the CTD,SwissTargetPrediction,GeneCards,and OMIM databases.After importing the targets into the STRING database,protein-protein interaction analysis was conducted to identify the key targets for metformin,osteoarthritis,and ferroptosis.(2)Molecular docking:P53 and its downstream factor SLC7A11 protein structures in PDB format were downloaded from the PDB database.The 2D structure of metformin was converted to a 3D structure,and molecular docking of metformin with the proteins was performed using Discovery Studio 2019 Client.(3)In vivo experiments:Thirty male SD rats were randomly divided into three groups(n=10).The blank group did not receive surgery.The osteoarthritis model was established using the modified Hulth method for the model and metformin groups.One day after the surgery,rats in the metformin group were gavaged with metformin 200 mg/kg per day,while the blank and model groups were gavaged with physiological saline.Treatment continued for 4 weeks.Hematoxylin-eosin staining and Safranin O-fast green staining were used to observe the pathological morphology and structure of the knee cartilage,and Mankin scoring was performed.ELISA was used to measure the levels of tumor necrosis factor-α and interleukin-6 in the serum.The microplate method was used to measure serum ferroptosis-related indicators,including glutathione,malondialdehyde,and Fe2+.Immunofluorescence staining,western blot assay,and real-time qPCR were used to detect the protein and mRNA expression of P53,SLC7A11,glutathione peroxidase 4,proteoglycans,and matrix metalloproteinase 13 in the cartilage tissue of the rats.RESULTS AND CONCLUSION:(1)A total of 96 intersecting targets among metformin,osteoarthritis,and ferroptosis were identified.After protein-protein interaction analysis,77 potential targets were found.Further screening identified the core targets as TP53,AKT1,JUN,interleukin-6,MYC,interleukin-1β,and tumor necrosis factor-α,among others.(2)Docking analysis results showed that metformin bound strongly and stably with P53 and its downstream factor SLC7A11.(3)In the model group,the knee cartilage surface was irregular,with cartilage tissue defects and reduced chondrocyte numbers.Compared to the model group,the knee cartilage structure damage in the metformin group was significantly improved,with a smoother cartilage surface and increased chondrocyte numbers.The Mankin score in the model group was significantly higher than that in the blank group,while the Mankin score in the intervention group was significantly lower than that in the model group.(4)Compared with the model group,the metformin group had significantly lower levels of tumor necrosis factor-α,interleukin-6,malondialdehyde,and Fe2+,and significantly higher glutathione levels.(5)Compared to the model group,the metformin group had significantly increased protein and mRNA expression of SLC7A11,glutathione peroxidase 4,and proteoglycans,and significantly decreased protein and mRNA expression of P53 and matrix metalloproteinase 13 in their cartilage tissue.(6)The results indicate that metformin can effectively improve cartilage damage in osteoarthritis rats and alleviate chondrocyte ferroptosis by inhibiting the aberrantly activated P53/SLC7A11/glutathione peroxidase 4 signaling pathway.This improvement in chondrocyte iron metabolism and lipid peroxidation response further reduces cartilage matrix degradation and prevents further cartilage damage and inflammatory response.

OBJECTIVE To establish a visualized loop-mediated isothermal amplification(LAMP)for rapid detec-tion of Klebsiella pneumoniae and evaluate the sensitivity and specificity of the detection method.METHODS To-tally4 specific LAMP primers were designed by targeting to rcsA gene of the K.pneumoniae.The reaction condi-tions(temperature,time)and the parameters of the reaction system were optimized,the reaction was made visu-alized by using chimeric fluorescent dye SYBR Green Ⅰ.The optimal reactions conditions and reaction systems were determined based on the results of visualization and agarose gel electrophoresis.The optimized conditions and systems were used to test the sensitivity by diluting DNA template on a 10-fold gradient,meanwhile,the specifici-ty of the method was evaluated by detecting K.pneumoniae and other ten species of common bacteria and fungi.RESULTS The optimal reaction temperature was 63 ℃ after the optimization,with the reaction time 35 min;the concentration of buffer solution in the reaction system was determined as 0.8×,with the concentration of magne-sium ion 8 mmol/L,the concentration of dNTPs 1.4 mmol/L,the ratio of internal to external primers 6∶1,the concentration of Bst DNA polymerase 0.32 U/μl,the concentration of betaine 0.75 mmol/L.For the test of sensi-tivity,the method could detect the template with the concentration of 1.5 ng/μl.For the test of specificity,only the detection of K.pneumoniae could display positive visualized result and positive electrophoretic band.CONCLUSIONS The visualized LAMP with high specificity and sensitivity for rapid detection of K.pneumoniae is successfully established,facilitating the observation of the detection result without the use of precise instruments.It is suitable for the detection in grass-root laboratories after successful preliminary application in clinical detection.

Objective:To evaluate the cumulative live birth rate (CLBR) and cost-effectiveness of fixed versus adjusted follicle-stimulation hormone (FSH) dosages in infertile women with different ovarian responses during their first assisted reproductive technology (ART) cycle.Methods:A retrospective real-world cohort study was conducted on 5 419 infertile women who underwent their first ART treatment at the Department of Reproductive Medicine of the First Affiliated Hospital of Nanjing Medical University between January 2013 and December 2017. All patients received an individualized starting dosage of gonadotropin. Based on whether FSH dosages were adjusted during controlled ovarian stimulation (COS), patients were divided into fixed-dosage group ( n=2 061) and adjusted-dosage group ( n=3 358). Clinical outcomes and FSH cost-effectiveness were compared between the two groups across different ovarian response groups, with CLBR as the primary outcome. Propensity score matching (PSM) and multivariable logistic regression were used to adjust for potential confounders. Results:FSH dosage adjustments were found in 62.0% (3 358/5 419) of cycles during COS. After PSM, baseline characteristics were comparable between the two groups (all P>0.05). After adjusting for confounders using multivariable logistic regression, FSH dosage adjustment was not significantly associated with CLBR ( OR=1.06, 95% CI: 0.94-1.20, P=0.332). Compared with the adjusted-dosage group, the fixed-dosage group showed no significant differences in CLBR in poor-, normal-, and high-responder groups (all P>0.05). The incidence of ovarian hyperstimulation syndrome (OHSS) did not differ significantly between the two groups ( P>0.05). In poor-, normal-, and high-responder groups, the total FSH dosages in the fixed-dose group [1 350 (375, 1 825) U, 1 200 (375, 1 500) U and 525 (375, 1 128) U, respectively] were significantly lower than those in the adjusted-dose group [1 875 (1 425, 2 294) U, P=0.001; 1 425 (450, 1 875) U, P<0.001; 600 (375, 1 425) U, P=0.020]. Similarly, average FSH costs in different ovarian response groups in the fixed-dosage group [4 725.0 (1 312.5, 6 387.5) yuan, 4 200.0 (1 312.5, 5 250.0) yuan and 1 837.5 (1 312.5, 3 947.3) yuan, respectively] were significantly lower than those in the adjusted-dosage group [6 562.5 (4 987.5, 8 028.1) yuan, P=0.001; 4 987.5 (1 575.0, 6 562.5) yuan, P<0.001; 2 100.0 (1 312.5, 4 987.5) yuan, P=0.020]. For normal-responders, the FSH cost per high-quality embryo in the fixed-dosage group [1 365.0 (875.0, 2 537.5) yuan] was significantly lower than that in the adjusted-dosage group [2 056.3 (1 268.8, 3 412.5) yuan, P<0.001]. Conclusion:FSH dosage adjustment during COS is not associated with CLBR or the incidence of OHSS. However, the fixed-dose group exhibited lower total FSH dosages and costs across different ovarian response populations. In the context of ART being covered by medical insurance, fixed FSH dosage may represent a more cost-effective ovarian stimulation protocol.

Objective To explore the targets and mechanisms of Tongguanteng Injection in inhibiting the proliferation and migration of cervical cancer.Methods The biological activity of Tongguanteng Injection in inhibiting human cervical cancer SiHa cells was determined by MTT method.Detecting the effect of Tongguanteng Injection on SiHa cell migration through wound healing assay.Using network pharmacology to collect the key targets for treating cervical cancer,and perform molecular docking and enrichment analysis on the targets.Immunohistochemistry and Western blot were used to detect the key proteins to validate the network pharmacology predictions.Result Tongguanteng Injection significantly inhibited the proliferation and migration in a dose-dependent manner in human cervical cancer SiHa cells.Based on the main active ingredients of Marsdenia tenacissima,81 therapeutic targets for cervical cancer were obtained,which may treat cervical cancer by affecting key proteins such as FGF2,MAPK1,and MAPK3.Immunohistochemical results indicated that FGF2,MAPK1 and MAPK3 were expressed in cervical cancer tissues.The western bolt assays showed that Tongguanteng Injection could significantly reduce the FGF2 protein expression.Meanwhile,the MAPK1 and MAPK3 protein expressions were significantly increased.Conclusion Tongguanteng Injection may regulate the FGF2,MAPK1 and MAPK3,effectively impede the proliferation and migration of cervical cancer.

Objective:To explore the expression of bone marrow stromal antigen 2(BST2)in advanced non-small cell lung cancer(NSCLC)tissues and its impact on the prognosis of cisplatin chemotherapy.Methods:This study was a prospective study,127 advanced NSCLC patients who received treatment at Shangrao People's Hospital from January 2022 to December 2023 were prospective selected,all patients received cisplatin+gemcitabine chemotherapy regimen.They were divided into survival group(n=85)and death group(n=42)according to the prognosis.The relationship between expression of BST2 and clinicopathological characteristics was analyzed.The survival situation was analyzed by Kaplan-Meier method.Influencing factors of prognosis after cisplatin chemotherapy were discussed by univariate and multivariate COX regression analyses.Results:The positive expression rate of BST2 in cancer tissues(34.65％)in advanced NSCLC tissues was significantly higher than that in adjacent tissues(7.09％)(P＜0.05).Expression of BST2 was associated with tumor TNM staging,lymph node metastasis and differentiation degree(P＜0.05).The 1-year survival rate of patients with high expression of BST2(56.98％)was significantly lower than that of patients with low expression(87.80％)(P＜0.05).High expression of BST2 and TNM stage Ⅳ were independent prognostic risk factors for cisplatin chemotherapy in patients with advanced NSCLC(P＜0.05).Conclusion:The expressed of BST2 is highly in patients with advanced NSCLC tissues and closely related to cisplatin chemotherapy poor prognosis,suggesting its potential as a prognostic biomarker.

Aim To investigate the effect and mechanism of angiotensin Ⅱ(Ang Ⅱ)on ferroptosis in white adi-pocytes.Methods The 3T3-L1 preadipocytes were differentiated into white adipocytes by inducer stimulation.The experiment was divided into control group,Ang Ⅱ group,Ang Ⅱ+Fer-1(ferroptosis inhibitor)group and Ang Ⅱ+PFT-α(p53 inhibitor)group.Ang Ⅱ was used to treat cells.RT-qPCR and Western blot were used to detect the expression levels of ferroptosis factors and adipokines.JC-1 kit was used to detect mitochondrial membrane potential(MMP)level.Iron ion kit was used to detect intracellular iron content.Glutathione(GSH)kit was used to detect GSH content.Fer-1 and Ang Ⅱ were added to treat cells to detect the the changes of ferroptosis level.The expression of p53 and spermidine/spermine N1-acetyltransferase 1(SAT1)protein was detected.Subsequently,PFT-α and Ang Ⅱ were added to co-treat cells to detect the changes of p53 and SAT1 protein expression,and to observe the effect of inhibiting p53 expression on the expression levels of ferroptosis factors and adipokines.Results 3T3-L1 cells were successfully differentiated into white adipocytes by stimulator-induced differentiation.Ang Ⅱ induced ferroptosis in white adipocytes.RT-qPCR results showed that compared with control group,the mRNA expression of anti-ferroptosis factor glutathione peroxidase 4(GPX4),solute carrier family 7 member 11(SLC7A11)and iron regulatory protein 1(IRP-1)was down-regulated in Ang Ⅱ group,and the mRNA expression of pro-ferroptosis factor acyl-CoA synthetase of long-chain family member 4(ACSL4)was up-regulated.Western blot results showed that compared with control group,the protein expression of SLC7A11 and GPX4 was down-regulated in Ang Ⅱ group,and the protein expression of ACSL4 was up-regulated.Ang Ⅱ treatment increased the content of intracellular iron ions and decreased the levels of GSH and MMP.Compared with Ang Ⅱ group,the mRNA expression of IRP-1 and SLC7A11 was up-regulated in Ang Ⅱ+Fer-1 group.Ang Ⅱ induced changes in the expression profile of adipokines in white adipocytes.Western blot results showed that compared with control group,the protein ex-pression of pro-inflammatory adipokine leptin(LEP),resistin(RETN),interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)was up-regulated in Ang Ⅱ group,and the protein expression of anti-inflammatory adipokine adiponectin(AD-PN)and omentin 1(ITLN1)was down-regulated.In addition,Ang Ⅱ increased the protein expression of p53 and SAT1.Inhibition of p53 expression can improve the level of ferroptosis and adipokine expression in white adipocytes trea-ted with Ang Ⅱ.Western blot results showed that compared with Ang Ⅱ group,the protein expression of p53 and SAT1 was down-regulated in Ang Ⅱ+PFT-α group,the protein expression of SLC7A11 and GPX4 was up-regulated,and the protein expression of ACSL4 was down-regulated.The protein expression of ADPN was up-regulated in Ang Ⅱ+PFT-αgroup,and the protein expression of TNF-α,LEP and RETN was down-regulated.Conclusion Ang Ⅱ induces fer-roptosis in white adipocytes through activating the p53/SAT1 signaling pathway.

OBJECTIVE To establish a visualized loop-mediated isothermal amplification(LAMP)for rapid detec-tion of Klebsiella pneumoniae and evaluate the sensitivity and specificity of the detection method.METHODS To-tally4 specific LAMP primers were designed by targeting to rcsA gene of the K.pneumoniae.The reaction condi-tions(temperature,time)and the parameters of the reaction system were optimized,the reaction was made visu-alized by using chimeric fluorescent dye SYBR Green Ⅰ.The optimal reactions conditions and reaction systems were determined based on the results of visualization and agarose gel electrophoresis.The optimized conditions and systems were used to test the sensitivity by diluting DNA template on a 10-fold gradient,meanwhile,the specifici-ty of the method was evaluated by detecting K.pneumoniae and other ten species of common bacteria and fungi.RESULTS The optimal reaction temperature was 63 ℃ after the optimization,with the reaction time 35 min;the concentration of buffer solution in the reaction system was determined as 0.8×,with the concentration of magne-sium ion 8 mmol/L,the concentration of dNTPs 1.4 mmol/L,the ratio of internal to external primers 6∶1,the concentration of Bst DNA polymerase 0.32 U/μl,the concentration of betaine 0.75 mmol/L.For the test of sensi-tivity,the method could detect the template with the concentration of 1.5 ng/μl.For the test of specificity,only the detection of K.pneumoniae could display positive visualized result and positive electrophoretic band.CONCLUSIONS The visualized LAMP with high specificity and sensitivity for rapid detection of K.pneumoniae is successfully established,facilitating the observation of the detection result without the use of precise instruments.It is suitable for the detection in grass-root laboratories after successful preliminary application in clinical detection.