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Live MSCs Characterizer Displays Stemness and Differentiation Using Colorful LV-cp Biosensors

ACS SENSORS 2025年第2期10卷 825-834页

作者： Jiang, Qingyun Shao, Shuai Li, Na zhang, zhengyao Zhao, Luming zhang, Hangyu Liu, Bo Canc Hosp Dalian Univ Technol Shenyang 110042 Peoples R China Dalian Univ Technol Fac Med Liaoning Key Lab Integrated Circuit & Biomed Elect Dalian 116024 Peoples R China Dalian Univ Technol Sch Chem Engn Ocean & Life Sci Panjin 124221 Peoples R China

Mesenchymal stem cells (MSCs) have garnered significant attention in biomedical research due to their accessibility and remarkable differentiation potential. However, the lack of efficient and convenient living cell monitoring methods limits their widespread application in tissue engineering and stem cell therapy. Therefore, we present progress in the development of a novel series of fluorescent protein (FP) sensors based on turn-on fluorescent protein biosensors (Turn-on FPBs), termed the LV-cp biosensor system (novel live cell permuted fluorescent protein biosensors). Utilizing phage display technology to screen for affinity peptides specifically targeting MSCs and chondrocytes, the LV-cp were engineered by subcloning these peptides into permuted fluorescent proteins, thereby integrating the fluorescence activation mechanism with the affinity peptides and achieving highly accurate detection and identification of these two cell types using living cells as "fluorescence keys." This system provides a simplified, nontoxic method to replace traditional antibody kits, and strong fluorescence signals can be obtained through various fluorescence detection devices. In addition, the LV-cp biosensors enabled dynamic observation of MSCs differentiation into chondrocytes through changes in the cell fluorescence colors.

关键词： turn-on fluorescent protein biosensors on-invasiveness mesenchymal stem cells differentiation dynamicmonitoring

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Visualization of Apical-Basal Stress Polarity Regulating Directed Cell Migration with a FRET-Based Biosensor

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ACS SENSORS 2025年第2期10卷 671-677页

作者： Wang, Cunli Xu, Dengyue Shao, Shuai Han, Jinxin Li, Na zhang, zhengyao zhang, Hangyu Liu, Bo Dalian Univ Technol Canc Hosp Shenyang 110042 Peoples R China Dalian Univ Technol Fac Med Sch Biomed Engn Liaoning Key Lab Integrated Circuit & Biomed Elect Dalian 116024 Peoples R China Dalian Univ Technol Sch Life Sci & Med Panjin 116024 Peoples R China

Intracellular morphological apical-basal polarity, regulated by conserved polarity proteins, plays a crucial role in cell migration and metastasis. In this study, using a genetically encoded Forster resonance energy transfer (FRET) biosensor to visually present the spatiotemporal stress state between the lipid rafts on the membrane and the linked actin, we first provide the evidence for the existence of intrinsic apical-basal stress polarity in tumor cells and demonstrate that this polarity is a prerequisite for the formation of flow-induced front-back stress polarity. Interestingly, our study revealed that the front-back stress polarity disappeared upon the disruption of intrinsic apical-basal stress discrepancy, resulting in a large attenuated cell migration activity reduced from 76.2 +/- 3.5% to 10 +/- 4.9%. This unexpected discovery not only highlights the significance of apical-basal stress polarity as another factor influencing cell migration but also indicates that the disruption of apical-basal polarity in stress might be a novel therapeutic approach for tumor metastasis.

关键词： cell membrane cytoskeleton stress polarity FRET biosensor cell migration

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Microstructure and corrosion behaviour of Mg-1Sc alloy with different processing conditions

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MATERIALS SCIENCE AND TECHNOLOGY 2025年第5期41卷 320-329页

作者： zhang, Dan zhang, Cheng Wang, Jialin Yang, Qingshan Peng, Peng Long, Shuai Li, Xianzheng Tan, Bochuan zhang, zhengyao Dai, Qingwei Chongqing Univ Sci & Technol Sch Met & Mat Engn Chongqing 401331 Peoples R China Chongqing Res Inst China Coal Technol Engn Grp Chongqing Peoples R China

The microstructure and corrosion resistance of Mg-1Sc alloys in different processing conditions were investigated. The composition was examined by Inductively Coupled Plasma Atomic Emission Spectrometer, the microstructure was analyzed by metallographic microscope. The corrosion behaviour of the alloy was investigated by hydrogen evolution tests and electrochemical tests. The results showed that as-cast and rolled Mg-1Sc possess better corrosion resistance compared with extruded Mg-1Sc alloys, which is due to the fact that the extrusion flow lines and residual stresses caused by machining in extruded Mg-1Sc alloys can lead to easier corrosion. In addition, rolled Mg-1Sc alloy shows better corrosion resistance than as-cast alloy, which is attributed to the finer grain sizes and a denser corrosion product film.

关键词： Mg-Sc processing conditions microstructure corrosion behaviour electrochemistry

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MB based RT-qPCR increase the clinical application of cfEBV DNA for NPC in non-endemic area of China

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SCIENTIFIC REPORTS 2025年第1期15卷 1-12页

作者： He, Qiao Tuo, Yi Zhou, Yi Yan, Yue Liu, Xin Zhao, Dan Wang, Qiuju Luo, Hao zhang, zhengyao Meng, Fanping Ying, Binwu Wang, Dongsheng Yang, Mu Huang, Yecai Univ Elect Sci & Technol China Sichuan Canc Hosp & Inst Sichuan Clin Res Ctr Canc Sichuan Canc CtrDept Clin LabSch Med Chengdu 610000 Peoples R China Sichuan Univ West China Hosp Dept Lab Med Chengdu 610041 Peoples R China Chengdu Univ Tradit Chinese Med Coll Med Technol Chengdu 611137 Peoples R China Chong Qing Univ Dept Clin Lab Three Gorges Hosp Chongqing Peoples R China Univ Elect Sci & Technol China Sichuan Canc Hosp & Inst Sichuan Clin Res Ctr Canc Sichuan Canc CtrDept Expt Res Chengdu 610000 Peoples R China Univ Elect Sci & Technol China Dept Radiat Oncol Radiat Oncol Key Lab Sichuan Prov Sichuan Clin Res Ctr CancSichuan Canc Hosp & Inst Chengdu 610000 Peoples R China Univ Elect Sci & Technol China Sichuan Canc Hosp & Inst Sichuan Canc Ctr Dept Clin LabSichuan Clin Res Ctr Canc Chengdu 610000 Sichuan Peoples R China

To compare the performance of magnetic bead (MB) and the concentrated precipitation (CP) based RT-qPCR to qualify cell free EBV DNA (cfEBV DNA) for nasopharyngeal carcinoma (NPC) in non-endemic area of China. From January 2014 to June 2024, a retrospective analysis of 2 cohort studies on cfEBV DNA in NPC patients was conducted to assess the diagnostic value, positive detection rate and clinical application. cfEBV DNA detection with CP based RT-qPCR in cohort 1 and MB based RT-qPCR method in cohort 2. The MB based RT-qPCR for the quantitative measurement of cfEBV DNA load was higher than the CP based RT-qPCR in the same plasma samples from NPC patients (P < 0.001). CP based RT-qPCR measured cfEBV DNA in 1405 NPC and 244 healthy controls in cohort 1 with 40.8% sensitivity (AUC = 0.704, 95% CI: 0.676-0.731). In cohort 2(683 naive NPC and 303 controls), cfEBV DNA had a sensitivity of 75.84% (AUC = 0.879, 95% CI: 0.86-0.90). There were no significant differences in TNM stage among NPC between the two cohorts (P > 0.05). The MB method considerably increased the positive detection rate of cfEBV DNA in NPCs at stages III-IV, T2-T4, N1-N3, and M0 (P < 0.001). At the end of treatment, 97.51% of patients had no detectable EBV and just 2.49% had detectable cfEBV DNA. Those who received <= 2 or >= 3 cycles of NAC had a median t(1/2) clearance rate of 9.8 days and 12.6 days, respectively. MB based RT-qPCR increased the quantity of cfEBV DNA. MB based RT-qPCR demonstrated superior sensitivity and positive detection rates for cfEBV DNA. cfEBV DNA can be more positively noticed, with a higher diagnostic value and a broader variety of clinical applications among NPC in non-endemic areas.

关键词： RT-qPCR MB CP cfEBV DNA NPC

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Integrated Device for Cancer Nucleic Acid Biomarker Detection at Body Temperature

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MICROMACHINES 2025年第2期16卷 192-192页

作者： Chen, Chang Wu, Bin Li, Xuesong Jin, Yuhang zhang, Hangyu Liu, Bo zhang, zhengyao Li, Na Dalian Univ Technol Cent Hosp Dalian 116033 Peoples R China Dalian Univ Technol Fac Med Dalian 116024 Peoples R China China Certificat & Inspect Grp Liaoning Co Ltd Dalian 116001 Peoples R China Dalian Univ Technol Liaoning Key Lab Integrated Circuit & Biomed Elect Dalian 116024 Peoples R China Dalian Univ Technol Sch Chem Engn Ocean & Life Sci Panjin 124221 Peoples R China

The quantitative analysis of nucleic acid markers is extensively utilized in cancer detection. However, it faces significant challenges, such as the need for specialized detection devices and the inherent complexity of testing procedures. To address these issues, this study proposes a simplified, rapid, and user-friendly platform for cancer nucleic acid marker detection. We firstly designed a polydimethylsiloxane (PDMS) device for the isothermal amplification reaction of nucleic acid biomarkers based on reverse-transcription recombinase-aided amplification (RT-RAA) technology. Specifically, three potential cancer nucleic acid biomarkers, carcinoembryonic antigen (CEA), prostate-specific antigen (PSA), and prostate cancer antigen 3 (PCA3) were amplified from human serum or urine samples in the PDMS device at body temperature. The reaction chamber was directly integrated with nucleic acid test strips labeled with colloidal gold nanoparticles, allowing for the visual observation of the detection results for the amplification products. The optimal reaction conditions, such as pH, reaction time, antibody, and streptavidin concentration, were defined after a series of optimization studies. The findings demonstrated that the optimal RT-RAA reaction time was 20 min, the primary antibodies were labeled with colloidal gold to the greatest extent at pH 8.5, and the optimal concentrations of secondary antibody and streptavidin were 1.0 mg/mL and 0.5 mg/mL, respectively. Furthermore, this novel detection approach could not only exhibit excellent sensitivity and specificity but also show high accuracy for the analysis of nucleic acid biomarkers in both clinical serum and urine samples. Therefore, the simplified and more convenient operation platform provides a new insight for the semi-quantitative analysis of cancer nucleic acid biomarkers and the rapid screening of early cancer, thereby offering a promising alternative to oncological point-of-care testing (POCT) diagnostics.

关键词： integrated device early cancer detection nucleic acid biomarkers RT-RAA colloidal gold test strip

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Impact of RIM-BPs in neuronal vesicles release

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BRAIN RESEARCH BULLETIN 2021年 170卷 129-136页

作者： Gao, Tianyu zhang, zhengyao Yang, Yunong zhang, Hangyu Li, Na Liu, Bo Dalian Univ Technol Sch Biomed Engn Liaoning Key Lab Integrated Circuit & Biomed Elec Dalian 116024 Peoples R China Dalian Univ Technol Sch Life & Pharmaceut Sci Panjin Campus Panjin 124221 Peoples R China

Accurate signal transmission between neurons is accomplished by vesicle release with high spatiotemporal resolution in the central nervous system. The vesicle release occurs mainly in the active zone (AZ), a unique area on the presynaptic membrane. Many structural proteins expressed in the AZ connect with other proteins nearby. They can also regulate the precise release of vesicles through protein-protein interactions. RIM-binding proteins (RIM-BPs) are one of the essential proteins in the AZ. This review summarizes the structures and functions of three subtypes of RIM-BPs, including the interaction between RIM-BPs and other proteins such as Bassoon and voltage-gated calcium channel, their significance in stabilizing the AZ structure in the presynaptic region and collecting ion channels, and ultimately regulating the fusion and release of neuronal vesicles.

关键词： RIM-binding proteins Synapse Active zone Vesicle release Voltage-gated calcium channel

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Comparative proteomics analysis of human osteosarcoma by 2D DIGE with MALDI-TOF/TOF MS

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JOURNAL OF BONE ONCOLOGY 2016年第4期5卷 147-152页

作者： Wang, Guoxiang zhang, zhengyao Yang, Maoguang Xu, Bo Gao, Qi Yang, Xiaoyu Jilin Univ Dept Orthoped Hosp 2 ZiQiang St Changchun 130041 Peoples R China Jilin Univ Dept Orthoped Hosp 1 XinMin St Changchun 130041 Peoples R China Jilin Univ Sch Pharm Dept Pharmaceut Chem FuJi Rd Changchun 130041 Peoples R China ChangChun Normal Univ Dept Biol Elementary Educ Coll ChangJi Rd Changchun 130041 Peoples R China Dalian Univ Technol Sch Life Sci & Med DaGong Rd Panjin 124221 Peoples R China ChangChun Cent Hosp Dept Med Adm RenMin St Changchun 130041 Peoples R China Jilin Univ Dept Endocrinol Hosp 2 ZiQiang St Changchun 130041 Peoples R China

Osteosarcoma (OS) is the most common primary malignant tumor of bone and the third most common cancer in childhood and adolescence. However, controversy concerning the ideal combination of chemotherapy agents ensued throughout the last quarter of the 20th century because of conflicting and often nonrandomized data. Collaborative efforts to increase understanding of the biology of osteosarcoma and the use of preclinical models to test novel protein targets will be critical to identify the path toward improving outcomes for patients. We attempted to identify potential protein markers or therapy targets of osteosarcoma and give a glance at tumorigenesis of osteosarcoma. A sensitive and accurate method was employed in comparative proteomic analysis between benign tumor and osteosarcoma. Tumor tissues obtained by open biopsy before induction chemotherapy were investigated With 2D DIGE and MALDI-TOF/TOF MS, 22 differentially expressed proteins were identified after database searching, including 8 up-regulated and 14 down-regulated proteins. We also validated the expression levels of interesting proteins(have higher Ratios(tumor/normal)) by Western blotting assay. Annotating by bioinformatic tools, we found structural and signal transduction associated proteins were in large percentage among altered level proteins. In particular, some low abundant proteins involving translation and transcription, such as EEF2(Elongation Factor 2), LUM Lumican 23 kDa Protein) and GTF2A2(Transcription Initiation Factor Iia Gamma Chain.), were firstly reported by our study comparing to previous observations. Our findings suggest that these differential proteins may be potential biomarkers for diagnosis or molecules for understanding of osteosarcoma tumorigenesis, coming with biologic, preclinical, and clinical trial efforts being described to improve outcomes for patients.

关键词： Proteomics Osteosarcoma Benign Tumor MALDI-TOF/TOF MS

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The Pathological Factors Involved in Current In Vitro Atherosclerotic Models

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TISSUE ENGINEERING PART B-REVIEWS 2024年第5期30卷 530-544页

作者： Bao, Yuxin zhang, Hangyu Wang, Danbo Yan, Peishi Shao, Shuai zhang, zhengyao Liu, Bo Li, Na Dalian Univ Technol Fac Med Sch Biomed Engn Dalian Peoples R China Dalian Univ Technol Canc Hosp Shenyang Peoples R China Dalian Univ Technol Dept Cardiol Cent Hosp Dalian Peoples R China Dalian Univ Technol Sch Life & Pharmaceut Sci Panjin Peoples R China Dalian Univ Technol Fac Med Sch Basic Med Sci Dalian Peoples R China Dalian Univ Technol Liaoning Key Lab Integrated Circuit & Biomed Elect Dalian Peoples R China Dalian Univ Technol Fac Med Sch Basic Med Sci NO2Linggong Rd Dalian 116024 Peoples R China

Cardiovascular disease stemmed from atherosclerosis (AS) is well recognized to be the predominant cause of global death. To comprehensively clarify the pathogenesis of AS, exploit effective drugs, as well as develop therapeutic solutions, various atherosclerotic models were constructed in vitro and widely utilized by the scientific community. Compared with animal models, the in vitro atherosclerotic models play a prominent role not only in the targeted research of single pathological factor related to AS in the human derived system, but also in the combined study on multipathological factors leading to AS, thereby contributing tremendously to the in-depth elucidation of atherosclerotic pathological process. In the current review, a variety of pathological factors incorporated into the existing atherosclerotic models in vitro are broadly elaborated, including the pathological mechanism, in vitro simulation approaches, and the desired improvement perspectives for reproducing each pathological factor. In addition, this review also summarizes the advantages and disadvantages of current atherosclerotic models as well as their potential functionality. Finally, the promising aspects for future atherosclerotic models in vitro with potential advances are also discussed.

关键词： atherosclerosis in vitro models pathological factors biochemical factors biomechanical factors

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Accurate detection of reactive oxygen species by tuning an elastic motif (GPGGA)4 in nanopores

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CHEMICAL COMMUNICATIONS 2023年第29期59卷 4368-4371页

作者： Wang, Cunli Yang, Yiming Shao, Shuai zhang, Hangyu Li, Na zhang, zhengyao Liu, Bo Dalian Univ Technol Sch Biomed Engn Liaoning Key Lab Integrated Circuit & Biomed Elect Lingshui Rd Dalian 116024 Peoples R China

We have developed a reactive oxygen species (ROS) sensor based on nanopores modified with GGGCEG(GPGGA)(4)CEG. The formation of an intramolecular disulfide bond oxidized by ROS leads to conformation changes in GGGCEG(GPGGA)(4)CEG, which then induces an obvious change in the size of the nanopores and a corresponding ionic current change. This work allows the accurate and dynamic monitoring of ROS through the combination of (GPGGA)(4) and nanopores.

关键词： Nanopores

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S100A4: a novel partner for heat shock protein 47 in antler stem cells and insight into the calcium ion-induced conformational changes

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JOURNAL OF BIOMOLECULAR STRUCTURE & DYNAMICS 2020年第7期38卷 2068-2079页

作者： Shang, Yudong zhang, zhengyao Ba, Hengxing Wang, Datao Qi, Xiaoyan Li, Chunyi Chinese Acad Agr Sci Inst Special Wild Econ Anim & Plants Changchun Jilin Peoples R China State Key Lab Mol Biol Special Econ Anim Changchun Jilin Peoples R China Dalian Univ Technol Sch Life Sci & Med Panjin Peoples R China

S100A4 is a multiple-function protein highly expressed in tumor or stem cells. We found S100A4 was a novel protein partner for heat shock protein 47 (HSP47) in deer antlerogenic periosteum cells (AP cells), indicating that S100A4 could bind with HSP47. S100A4 had both calcium-dependent and calcium-independent patterns (labeled as SCd and SCi, respectively) to execute different biological activities. Homology models of HSP47, SCd and SCi were constructed. HSP47:collagen model, HSP47:collagen I-V, HSP47:SCd and HSP47:SCi complexes were built using ZDOCK software. Together with free SCd and SCi, 200 ns molecular dynamic (MD) simulations were performed to analyze binding free energies and SCi/SCd conformational changes. The energetic results showed that SCi had the strongest affinity to HSP47, and followed by collagens. SCd had little interaction with HSP47. Decomposition energy results showed that collagen model interacted with HSP47 mainly though neutral amino acids. When SCi bound with HSP47, the majority of mediated amino acids were charged. These results indicated that SCi could compete with collagen on the binding site of HSP47. Root mean square fluctuation (RMSF) values and cross-correlation matrices of principal component analysis (PCA) were calculated to evaluate the SCi/SCd structural variation during MD simulation. Both HSP47 and Ca2+ could stabilize the conformation of SCi/SCd. The loops interacting with Ca(2+)s and linking the two EF-hand motifs were impacted particularly. The relative moving directions of alpha-helices in EF-hands were distinct by the binding effect of HSP47 and Ca2+. We found that SCi may regulate the differentiation of AP cells by disturbing the interaction between HSP47 and collagen.

关键词： HSP47 protein-protein docking molecular dynamics antler stem cell

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