1.Research Progress of Metal-Organic Frameworks-Aptasensors for Detection of Contaminants in Food and Medicine Homology Substances
Xing GUO ; Jin-Ju TIAN ; Xiao-Zhen TANG ; Xiao-Yue WANG ; Na SONG ; Jin-E WANG ; Chao ZHU
Chinese Journal of Analytical Chemistry 2025;53(4):547-560
In recent years,the market share of food and medicine homology substances has continued to grow,and various types of contamination issues have become the focus of attention both inside and outside the industry.The contamination not only affects the original medicinal quality,but also leads to the accumulation of toxic substances in the human body,causing acute and chronic severe hazards such as vomiting,poisoning and cancer.Therefore,the development of biosensors that can conveniently,accurately and sensitively detect various pollutants in food and medicine homology substances has become a research hotspot.Aptasensors based on metal-organic frameworks(MOFs)with advantages such as strong specificity,rapid response and simple operation,have been widely used in detection of various pollutants.This review focused on the research progress of aptasensors based on MOFs for detection of food and medicine homology contamination in the past few years,and provided a detailed comparison and analysis for detection of chemical pollutants(such as pesticide residues,heavy metal residues,mycotoxins,etc.)and microbial contamination in food and medicine homology substances.Besides,the development trend and possible challenges of MOFs aptasensors in detection of food and medicine homology substances in the future were discussed,which was anticipated to provide a reference for the development of new MOFs aptasensors.
2.Screen of Disulfidptosis-related Colorectal Cancer Diagnostic and Therapeutic Target:Integrated Single-cell and Bulk RNA Sequencing Data
Yang YANG ; Yi-Xuan MA ; Xin-Yue FAN ; Wen-Xue ZHAO ; Yi-Ming QI ; Ning GAO ; Ju-Mei ZHAO ; Juan DU
Chinese Journal of Biochemistry and Molecular Biology 2025;41(10):1529-1540
Inflammatory response,immunosuppression,and drug sensitivity have been reported to have a significant correlation with the disulfidptosis levels in cancer patients.However,the value of disulfidpto-sis in colorectal cancer therapy remains unclear.Therefore,we classified the CRC cells into different cell types using single-cell sequencing data and cell-specific markers and analyzed their relationship with the cell disulfidptosis level.We found that the high disulfidptosis regions were concentrated in epithelial-like CRC cells.Further exploration using the disulfidptosis and programmed cell death 1 inhibitor therapy treated differential expression genes indicated that CRC patients with high disulfidptosis levels exhibited a lower risk profile and increased sensitivity to immunotherapy.By using the spatial transcriptomic analy-sis,we found that ubiquinol-cytochrome c reductase core protein 1(UQCRC1),a disulfidptosis-related gene,is highly expressed in epithelial-like CRC cells and co-localized with immune-infiltrated tumor re-gions.Additional bioinformatic analyses and experimental validation further confirmed that UQCRC1 was downregulated in CRC tissues.Overexpression of UQCRC1 suppressed CRC cell proliferation and migra-tion.These findings indicate that UQCRC1 is a potential target for CRC diagnosis and treatment.
3.Effects of High Temperature on Antioxidant,Carbon Metabolism Physiological Indicators and Transcriptome of Mytilus coruscus
Zhi YANG ; Yuan-Yue ZHANG ; Jian-Xin WANG ; Zhi LIAO ; Qun-Ju HU ; Qiao-Mei YANG ; Xiao-Lin ZHANG ; Mei-Hua FAN
Chinese Journal of Biochemistry and Molecular Biology 2025;41(3):446-459
The ocean plays a critical role in the global carbon cycle,and base on the"dual carbon"goals,ocean carbon sinks have received widespread attention.Shellfish aquaculture is one of the most important sources of carbon sinks in fisheries,which has an important impact on the offshore carbon cy-cle.As global temperature rises and ocean acidification intensifies,the capacity of the ocean to absorb CO2 will change.However,the effects of high temperature on the physiology and transcriptome related to carbon metabolism in Mytilus coruscus are not clear enough.This study investigated the effects of high temperatures on the total carbon content,carbon metabolism,antioxidant-related enzyme activities,and the transcriptome of Mytilus coruscus.The results showed that high temperature significantly inhibited the activities of hexokinase and pyruvate kinase,increased carbonic anhydrase activity(P<0.05),de-creased the ATP content of digestive glands(P<0.05),and affected glycolysis and the tricarboxylic acid cycle,leading to a significant decrease in the mussel's ability to sequester carbon.High temperature re-sulted in significant(P<0.05)increases in the levels of reactive oxygen species and malondialdehyde,and enhanced the activities of superoxide dismutase and catalase.Observations by transmission electron microscopy showed that high temperatures damaged the subcellular structure of the digestive gland in Mytilus coruscus,resulting in the shrinkage of the nucleolus,swelling of the endoplasmic reticulum,and a significant reduction in the mitochondrial cristae.Comparative transcriptomic analysis showed that the upregulated DEGs were mainly enriched in protein processing in the endoplasmic reticulum,antigen pro-cessing and presentation,and MAPK signaling pathway.The downregulated DEGs were mainly enriched in necroptosis,DNA replication,and the NF-kappa B signaling pathway.In antioxidant-related DEGs,the upregulated DEGs include vitamin K epoxide reductase,peroxidases,heat shock protein 105 kD,heat shock protein 70 kD,and superoxide dismutase;The downregulated DEGs mainly included NADPH oxidase,glutathione reductase,cytochrome b-245,cytochrome P450,and quinone reductase.The up-regulated genes enriched in the carbon metabolism pathway included chitinase,phosphatidylinositol 4,5-bisphosphate 3-kinase,phosphoenolpyruvate carboxykinase,galactokinase,and inositol trisphosphate 3-kinase.The downregulated genes included aldose-1-epimerase,carbonic anhydrase,galactose mutaro-tase,acyl-CoA synthetase,alcohol dehydrogenase,and hexokinase.In conclusion,high temperature has an inhibitory effect on the activities of enzymes and the expression of genes related to carbon metabolism in Mytilus coruscus.The results of this study are intended to provide a scientific basis for the healthy de-velopment of mussel aquaculture and the assessment of carbon sinks.
4.Effect and mechanism of LINC00839 on the malignant biological behavior of endometrial cancer cells
Yuan-yuan SHI ; Fen TIAN ; Wei-yue ZHOU ; Mei-yan LI ; Jian-cai MA ; Ju-rong WANG
Journal of Regional Anatomy and Operative Surgery 2025;34(1):21-27
Objective To investigate the effect of LINC00839 on the malignant biological behavior of endometrial cancer cells by regulating the miR-625-5p/cytoplasmic polyadenylation element binding protein 4 (CPEB4) axis. Methods The cancer tissues and adjacent tissues of 46 patients with endometrial cancer were obtained,endometrial cancer cells of HEC-1B,Ishikawa and RL95-2 and human endometrial epithelial cells (HEEC) were cultured in vitro,and the expression levels of LINC00839,miR-625-5p and CPEB4 in tissues and cells were detected. HEC-1B cells were divided into the HEC-1B group (conventional culture),sh-Ctrl group (transfected with sh-Ctrl),sh-LINC00839 group (transfected with shRNA-LINC00839),anti-miR-625-5p group (transfected with shRNA-LINC00839 and miR-625-5p inhibitor),and anti-NC group (transfected with shRNA-LINC00839 and inhibitor-NC). The proliferation,apoptosis,migration and invasion abilities of HEC-1B cells in each group were compared,and the expressions of CPEB4 and epithelial-mesenchymal transition (EMT) related proteins were determined. The targeting relationships between LINC00839 and miR-625-5p,and miR-625-5p and CPEB4 were analyzed. Results The mRNA expression of LINC00839 and CPEB4,as well as the positive expression rate of CPEB4 protein in endometrial cancer tissues were higher than those in adjacent tissues,and the expression of miR-625-5p was lower than that in adjacent tissues (P<0.05). Compared with HEEC cells,the expression of LINC00839 and the mRNA and protein expression of CPEB4 in Ishikawa,RL95-2 and HEC-1B cells increased (P<0.05),and the expression of miR-625-5p decreased (P<0.05). Compared with the sh-LINC00839 group and anti-NC group,the protein expression of N-cadherin,Vimentin,and CPEB4,24-hour absorbance,and migration and invasion cell numbers of HEC-1B cells in the HEC-1B group,sh-Ctrl group and anti-miR-625-5p group increased (P<0.05),while the expression of E-cadherin protein and cell apoptosis rate decreased (P<0.05). There were binding sites between LINC00839 and miR-625-5p,miR-625-5p and CPEB4,with targeting regulatory relationships. Conclusion LINC00839 is related to the malignant biological behavior of endometrial cancer cells,interference with LINC00839 expression can inhibit the proliferation,invasion and migration,and promote apoptosis of HEC-1B cells,and its mechanism may be achieved by regulating the miR-625-5p/CPEB4 axis.
5.Effects of High Temperature on Antioxidant,Carbon Metabolism Physiological Indicators and Transcriptome of Mytilus coruscus
Zhi YANG ; Yuan-Yue ZHANG ; Jian-Xin WANG ; Zhi LIAO ; Qun-Ju HU ; Qiao-Mei YANG ; Xiao-Lin ZHANG ; Mei-Hua FAN
Chinese Journal of Biochemistry and Molecular Biology 2025;41(3):446-459
The ocean plays a critical role in the global carbon cycle,and base on the"dual carbon"goals,ocean carbon sinks have received widespread attention.Shellfish aquaculture is one of the most important sources of carbon sinks in fisheries,which has an important impact on the offshore carbon cy-cle.As global temperature rises and ocean acidification intensifies,the capacity of the ocean to absorb CO2 will change.However,the effects of high temperature on the physiology and transcriptome related to carbon metabolism in Mytilus coruscus are not clear enough.This study investigated the effects of high temperatures on the total carbon content,carbon metabolism,antioxidant-related enzyme activities,and the transcriptome of Mytilus coruscus.The results showed that high temperature significantly inhibited the activities of hexokinase and pyruvate kinase,increased carbonic anhydrase activity(P<0.05),de-creased the ATP content of digestive glands(P<0.05),and affected glycolysis and the tricarboxylic acid cycle,leading to a significant decrease in the mussel's ability to sequester carbon.High temperature re-sulted in significant(P<0.05)increases in the levels of reactive oxygen species and malondialdehyde,and enhanced the activities of superoxide dismutase and catalase.Observations by transmission electron microscopy showed that high temperatures damaged the subcellular structure of the digestive gland in Mytilus coruscus,resulting in the shrinkage of the nucleolus,swelling of the endoplasmic reticulum,and a significant reduction in the mitochondrial cristae.Comparative transcriptomic analysis showed that the upregulated DEGs were mainly enriched in protein processing in the endoplasmic reticulum,antigen pro-cessing and presentation,and MAPK signaling pathway.The downregulated DEGs were mainly enriched in necroptosis,DNA replication,and the NF-kappa B signaling pathway.In antioxidant-related DEGs,the upregulated DEGs include vitamin K epoxide reductase,peroxidases,heat shock protein 105 kD,heat shock protein 70 kD,and superoxide dismutase;The downregulated DEGs mainly included NADPH oxidase,glutathione reductase,cytochrome b-245,cytochrome P450,and quinone reductase.The up-regulated genes enriched in the carbon metabolism pathway included chitinase,phosphatidylinositol 4,5-bisphosphate 3-kinase,phosphoenolpyruvate carboxykinase,galactokinase,and inositol trisphosphate 3-kinase.The downregulated genes included aldose-1-epimerase,carbonic anhydrase,galactose mutaro-tase,acyl-CoA synthetase,alcohol dehydrogenase,and hexokinase.In conclusion,high temperature has an inhibitory effect on the activities of enzymes and the expression of genes related to carbon metabolism in Mytilus coruscus.The results of this study are intended to provide a scientific basis for the healthy de-velopment of mussel aquaculture and the assessment of carbon sinks.
6.Qualitative study on risk perception of additional injury during motor rehabilitation in stroke patients with hemiplegia
Yue JU ; Qing WANG ; Jiachen ZHANG ; Sunling CONG ; Yun MAO ; Hui XU
Chinese Journal of Modern Nursing 2025;31(23):3097-3103
Objective:To explore the characteristics and influencing factors of risk perception related to additional injury during motor rehabilitation in stroke patients with hemiplegia, and to provide a foundation for the development of assessment tools and prevention strategies.Methods:A purposive sampling strategy combined with maximum variation sampling was employed to select 15 stroke patients with hemiplegia who were hospitalized in the Department of Neurology and Department of Rehabilitation at Northern Jiangsu People's Hospital between January and April 2024. Face-to-face, semi-structured in-depth interviews were conducted. Data were analyzed and thematically extracted using Colaizzi seven-step method.Results:Each interview lasted between 25 and 47 minutes, totaling 547 minutes of interview time and approximately 92 000 words of transcription. A total of 3 major themes were identified: characteristics of risk perception regarding additional injury during motor rehabilitation, multidimensional influencing factors of risk perception, and perceived severity of additional injury.Conclusions:Stroke patients with hemiplegia exhibit diverse and dynamic risk perception characteristics regarding additional injury during motor rehabilitation, which are influenced by multiple interrelated factors. A generally low and inaccurate level of risk perception was observed. Medical staff should pay greater attention to improving the accuracy of patients' risk perception, enhancing risk assessment mechanisms, and developing targeted prevention strategies to help patients avoid additional injuries and maximize the benefits of motor rehabilitation.
7.Usefulness of intraoperative choledochoscopy in laparoscopic subtotal cholecystectomy for severe cholecystitis
Rui-Hui ZHANG ; Xiang-Nan WANG ; Yue-Feng MA ; Xue-Qian TANG ; Mei-Ju LIN ; Li-Jun SHI ; Jing-Yi LI ; Hong-Wei ZHANG
Annals of Hepato-Biliary-Pancreatic Surgery 2025;29(2):192-198
Laparoscopic subtotal cholecystectomy (LSC) has been a safe and viable alternative to conversion to laparotomy in cases of severe cholecystitis. The objective of this study is to determine the utility of intraoperative choledochoscopy in LSC for the exploration of the gallbladder, cyst duct, and subsequent stone clearance of the cystic duct in cases of severe cholecystitis. A total of 72 patients diagnosed with severe cholecystitis received choledochoscopy-assisted laparoscopic subtotal cholecystectomy (CALSC). A choledochoscopy was performed to explore the gallbladder cavity and/or cystic duct, and to extract stones using a range of techniques. The clinical records, including the operative records and outcomes, were subjected to analysis. No LSC was converted to open surgery, and no bile duct or vascular injuries were sustained. All stones within the cystic duct were removed by a combination of techniques, including high-frequency needle knife electrotomy, basket, and electrohydraulic lithotripsy. A follow-up examination revealed the absence of residual bile duct stones, with the exception of one common bile duct stone, which was extracted via endoscopic retrograde cholangiopancreatography. In certain special cases, CALSC may prove to be an efficacious treatment for the management of severe cholecystitis. This technique allows for optimal comprehension of the situation within the gallbladder cavity and cystic duct, facilitating the removal of stones from the cystic duct and reducing the residue of the non-functional gallbladder remnant.
8.Study on the stability of PBMCs recovered from leukocyte-depleted filter residues
Ju LIN ; Zhiqiang XIANG ; Dongfen DU ; Fang YUAN ; Miaoyu WANG ; Yue WU ; Kaiyu HUANG ; Lieyong SANG
Chinese Journal of Blood Transfusion 2025;38(5):723-733
Objective: To identify an optimal back-flush solution for leukocyte-depleted filters that maximizes peripheral blood mononuclear cell (PBMC) recovery with high viability, long-term storage stability, and sterility of the harvested residues, thereby providing a clinically translatable strategy. Methods: Three sterile bag-packaged solutions—Saline, Solvent, and Hanks' balanced salt solution (HBSS)—were used to back-flush randomly assigned leukocyte-depleted filters. Nucleated cell recovery rate and viability of the harvested residues were compared. The optimal solution identified was applied to an expanded sample set. PBMC viability and yield were evaluated after 1h vs 48h storage of the residues. PBMCs isolated from the residues were cryopreserved in liquid nitrogen for 1 month, followed by post-thaw comparisons of viability and T-cell expansion capacity. Results: The Solvent group achieved the highest and most consistent nucleated cell recovery rate. Post-flush recovery rate from filters after 400 mL whole blood processing was (21.3±1.6)% for the Solvent group, significantly higher than Saline group (19.2±6.3)% and HBSS group (11.2±5.0)%, with residues from all groups maintaining viability >90%. No biologically significant difference in residue viability was observed between 48h vs 1h storage groups (93.3±2.3)% vs (95.7±1.8)%). PBMC recovery rates from residues showed no statistical difference between 48h vs 1h storage groups [(48.2%±9.5%)vs (40.41%±8.35%), P>0.05], with (17.7±2.6)×10
cells. After 1-month cryopreservation and 10-day expansion, PBMCs isolated from 48-hour-stored residues retained (91.2±3.2)% viability and achieved a (61.9±15.9)-fold expansion. Conclusion: The bag-packaged Solvent, as a back-flush solution, enables sterile acquisition of leukocyte-depleted filter residues through closed-system tubing connections. These residues maintained PBMC viability and recovery rates after 48h storage at 2℃-8℃, with post-cryopreservation (1-month liquid nitrogen) viability and expansion capacity remaining stable. This protocol complies with blood bank regulatory criteria, addresses the concerns about the infectious window period in cell therapy raw materials, and provides a clinically translatable strategy for PBMC-based applications.
9.Functional characterization of double-negative T cells isolated from leukoreduction filter residues.
Zhiqiang XIANG ; Yue WU ; Kaiyu HUANG ; Fuqiang WU ; Ju LIN ; Lieyong SANG ; Liming YANG
Journal of Zhejiang University. Medical sciences 2025;():1-9
OBJECTIVES:
To characterize the biological properties of double-negative T (DNT) cells isolated from leukoreduction filter residues.
METHODS:
Leukoreduction filters containing residues from 400 mL whole blood units (n=6) were collected from a blood center. Filters were back-flushed with normal saline, and the eluate was concentrated to obtain leukoreduction filter residues. Leukocytes in the residues were counted by dual-fluorescence staining. DNT cells were then isolated from the residues using antibody-mediated adsorption and density gradient centrifugation. Both cryopreserved and fresh unstimulated DNT cells derived from the residues were subjected to in vitro culture. Following culture, cells were assessed for expansion fold, viability, immunophenotype, differentiation status, and cytotoxicity against target cells using dual-fluorescence staining and flow cytometry, with comparisons made to DNT cells derived from whole blood.
RESULTS:
The leukocyte recovery rate achieved through reverse flushing of the leukocyte reduction filter was (41.9±14.7)%. Compared to whole blood, the DNT cell starting material obtained from filter residues showed no significant difference in total T-cell content (P>0.05). However, the viability and purity of the resulting DNT cell starting materials were significantly lower (both P<0.05). After 17 days of culture, DNT cells from filter residues and whole blood showed no significant differences in expansion fold, immunophenotype, differentiation status, or cytotoxicity toward target cells (all P>0.05). However, the viability of DNT cells from residues was significantly lower than that of whole blood-derived DNT cells [(86.0±4.2)% vs. (92.2±1.2)%, P<0.05]. After thawing (post 3 or 15 days of cryopreservation) and 17 days of culture, DNT cell starting materials from residues showed comparable immunophenotype, expansion fold, and differentiation status to their non-cryopreserved counterparts from the same source (all P>0.05). However, the viability of DNT cells cryopreserved for 3 days [92.4% (91.8%, 92.8%)] and the cytotoxicity against target cells of those cryopreserved for 15 days [91.3% (89.4%, 95.1%)] were significantly higher than those of non-cryopreserved DNT cells [87.8% (82.0%, 89.0%) and 70.9% (67.3%, 80.2%), respectively] (P<0.05).
CONCLUSIONS
DNT cells derived from leukoreduction filter residues exhibited highly comparable characteristics to those from whole blood in terms of expansion, purity, differentiation, and biological potency. Furthermore, their biological activity post-cryopreservation and revival remained largely similar to non-cryopreserved cells. These findings suggest that leukoreduction filter residues represent a promising alternative source of starting material for manufacturing off-the-shelf, allogeneic DNT cell therapeutics.
10.Serological and Molecular Biological Detection of RhD Variants.
Dao-Ju REN ; Chun-Yue CHEN ; Xiao-Wei LI ; Jun XIAO ; Xiao-Juan ZHANG ; Cui-Ying LI
Journal of Experimental Hematology 2025;33(2):498-503
OBJECTIVE:
To analyze the RHD genotyping and sequencing results of RhD serology negative samples in the clinic, and to further explore the laboratory methods for RhD detection, in order to provide a basis for clinical precision blood transfusion.
METHODS:
A total of 27 200 whole blood samples were screened for RhD blood group antigen using microcolumn gel card method.Serologic RhD-negative confirmation tests were performed on blood samples that were negative for RhD on initial screening using three different clonal strains of IgG anti-D reagents. The 10 exons of the RHD gene on chromosome 1 were also analyzed by PCR-SSP to determine RHD genotyping.When the PCR-SSP method did not yield definitive results, the RHD gene of the sample was analyzed by the third-generation sequencing.
RESULTS:
The results of the initial screening test by the microcolumn gel card method showed that 136 of the 27 200 samples were RhD-negative, of which 86 underwent RhD-negative confirmation testing and RHD genotyping, 88.37% (76/86 cases) of the RhD-negative confirmation test results were negative for the three anti-D reagents, and the results of RHD genotyping showed that 67.44% (58/86 cases) of the cases had a complete deletion of 10 exons, and the remaining 28 cases were RHD*711delC (1 case), RHD*D-CE(1-9)-D (1 case), RHD*D-CE(2-9-)D (2 cases), RHD*D-CE(3-9)-D (4 cases), RHD*DEL1 (c.1227G >A) mutation (16 cases), RHD*weak partial 15(845G >A) mutation (3 cases), and a mutation of c.165C >T base was found in 1 sample by three-generation sequencing.
CONCLUSION
RHD genotype testing of samples that are serologically negative for RhD antigen shows that some of the samples have RHD gene variants, not all of which are total deletions of RHD, suggesting that there are some limitations of the serologic method for RhD detection. Due to the polymorphism of the RHD gene structure, different RhD variants present different serologic features, which need to be further detected in combination with molecular biology testing, especially for the identification of Asian-type DELs, which is important for clinical precision blood transfusion.
Humans
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Rh-Hr Blood-Group System/genetics*
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Genotype
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Polymerase Chain Reaction
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Exons
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Blood Grouping and Crossmatching

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