CURRENT MOLECULAR MEDICINE
·
2018
·Vol. 18
·期 6
·383-391
·
IF 2.254
Li Yue,Lin Yan,Liufu Chun,Tu Jiawei,Yang Chaopin,Chen ZhiYi
Objective: To study transfection efficiency of folate-modified chitosan (FACS) nanoparticles as a non-viral vector delivering pEGFP-C3plasmid (FA-CS/P) to 293T cells with or without the combination of ultrasound and microbubble. Method: pEGFP-C3 was used as a reporter gene and
FA-CS nanoparticles, which were prepared by complex coagulation method, were used as biological carriers. Transfection efficiency to 293T cells mediated by FA-CS/P nanoparticles, ultrasound (US) and microbubble (MB) was assessed by fluorescence microscopy and flow cytometry. Result: FA-CS/P
nanoparticles have a particle size of 355.1 nm and zeta potential of 10.4 mV. Significant green fluorescence could be observed in CS/P group, FA-CS/P group, US+MB/P group, US+FA-CS/P group, Liposome 2000 (L) group under an inverted fluorescence microscope, while for US+MB+FA-CS/P group, only
scattered fluorescence was observed. Result of flow cytometry showed that the transfection rate of US+MB+FA-CS/P group was (2.0±0.2)%, which was significantly lower than other groups (P<0.05). CCK-8 experiments showed that cell vitality of US+MB+FA-CS/P was groups (P<0.05). CCK-8
experiments showed that cell vitality of US+MB+FA-CS/P was (64.1±4.6)%, which was also lower than other groups (P<0.05). Conclusion: In this study, FA-CS was successfully synthesized. FA-CS could combine with pEGFP-C3 effectively forming nanoparticles with nanoparticle size,
well dispersion, high encapsulation efficiency and no significant toxicity to cells. The application of ultrasound increased the transfection rate of FA-CS/P. However, while being exposed to ultrasound and microbubble, the transfection rate of FA-CS/P decreased obviously, may indicate that
there was no synergistic effect for gene transfection by the combination of ultrasound, folate modified chitosan and microbubbles.
研究领域:
生物医学工程
·
分子生物学
✓ 已验证
DOI
PubMed
高影响因子
LANGMUIR
·
2020
·
IF 3.557
Yiqun Wang,Yuan Yao
In natural tissue, nanofibrils in extracellular matrix (ECM), such as collagen fibrils, direct cell migration through contacting guidance. The contacting nanofibers on cell-ECM interface are reorganized from curl fibers to straightened fibers. However, how these nanofibers regulate single cell migration remains obscure. To investigate this issue, we fabricated collagen/polymer based biomimetic nanofiber sheets of varying topography. And we selected tumorigenic cell KGN and nontumorigenic cell 293T for comparison. We found KGN showed higher sensitivity to the nanofiber alignment rather than the nontumorigenic cell 293T, in morphological change, trajectory adaptation, and velocity variation. We also found aligned nanofibers shaped both KGN and 293T into elongated spindle morphology. Comparatively, KGN had greater perimeter and lower roundness than 293T. To study the dynamics of single cell migration of KGN and 293T, we conducted trajectory tracking and siRNA validation on regulatory proteins. We found nanofibers of varying topography regulated the patterns of single cell migration differently. For KGN cell, β-catenin, Rac1, and Cdc42 participated in its directional migration, but it was impervious to vimentin. Comparatively, epithelial cell 293T involved vimentin in its directional migration.
研究领域:
生物材料
·
细胞生物学
·
组织工程
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产品关联
DOI
PubMed
高影响因子
ACTA BIOCHIMICA ET BIOPHYSICA SINICA
·
2020
·Vol. 52
·期 5
·563-569
·
IF 2.836
Wang Lianqing,Lan Yongting,Du Yanqin,Xiang Xinxin,Tian Wenxiu,Yang Baoye,Li Tao,Zhai Qiaoli
Osteoblast differentiation is a key process in bone homeostasis. Mutations in plastin 3 have been reported to be responsible for X-linked osteoporosis. Plastin 3 and plastin 2 act synergistically to regulate osteoblast differentiation. However, the bone-related function of plastin 1, another family member of plastins, has not been assessed. In this study, we addressed the functional importance of plastin 1 in osteoblasts. We characterized the expression patterns of plastin 1 during osteoblast differentiation and revealed its important role in this process. In both HEK 293T and hFOB1.19 cells, plastin 1 was demonstrated to regulate intracellular Ca2+. Accordingly, we revealed that higher Ca2+ concentration promotes osteoblast differentiation. Finally, we found that plastin 1 may play a compensatory role in osteoporosis patients with plastin 3 deficiency. Together, our results indicate that plastin 1 promotes osteoblast differentiation by regulating intracellular Ca2+. Our work sheds new light on the role played by plastins in bone homeostasis.
研究领域:
骨骼生物学
·
细胞生理学
·
分子生物学
·
医学遗传学
✓ 已验证
DOI
PubMed
高影响因子
CHEMISTRY OF NATURAL COMPOUNDS
·
2025
·
IF 0.9
Wang Yong,Wang Wei,Liu Li-Na,Wei Dong-Lin,Liu Cong-Jun,Wei Shang-Hua,Gao Song,Yu Shu-Ping,Yao Ning,Wang Yu-Fei
A series of abietic acid derivatives modified with a triazole moiety at the C-18 position were synthesized using Huisgen 1,3-dipolar cycloaddition reaction of 18-azidoabietane with alkynes. The antitumor activity of the synthesized compounds was investigated through human cervical cancer cells (HeLa), human breast cancer cells (MCF-7) and human renal epithelial cells (HEK-293T). The outcome indicated that compounds 5e , 5f , 5j exhibited positive inhibitory activity against the tested tumor cell lines. In particular, the IC 50 value for the inhibitory activity of compound 5e against human breast cancer cells (MCF-7) was 11.2 μmol/L.
研究领域:
药物化学
·
有机合成
·
肿瘤药理学
·
天然产物衍生物
✓ 已验证
DOI
高影响因子
CHEMISTRY OF NATURAL COMPOUNDS
·
2024
·Vol. 60
·期 1
·92-97
·
IF 0.8
Wang Yong,Wang Wei,Liu Cong-Jun,Gao Song,Xiong Yong-Ge,Gao Tian-Zeng,Li Wei-Shi,Li Jing-Jing
Eight new stigmasterol derivatives possessing 1,2,3-triazoles moiety were synthesized by a 1,3-dipolar cycloaddition reaction. All the compounds were characterized by IR, NMR, and HR-MS. In addition, human cervical cancer cells (HeLa), human gastric cancer cells (HGC-27), and human renal epithelial cells (HEK-293T) were selected to study the antitumor activity of the synthesized compounds. The results demonstrated that compounds 3c and 3d exhibited better inhibitory activity against the tested tumor cell lines. The IC 50 value of the inhibitory activity on HeLa was 52.3 μmol·L –1 and 31.4 μmol·L –1 , and 40.6 μmol·L –1 and 44.6 μmol·L –1 for HGC-27, respectively.
研究领域:
有机合成
·
药物化学
·
药理学
·
癌症研究
·
细胞生物学
✓ 已验证
DOI
高影响因子
ANALYTICAL CHEMISTRY
·
2022
·
IF 8.008
Lancheng Wang,Mingjie Xu,Huimin Zhou,Kun Yan,Shiqi Duan,Dandan Xue,Youmei Wang,Bin Di,Chi Hu
The target of typical PCR analysis is restricted to nucleic acids. To this end, we report here a novel strategy to simultaneously detect genetic and metabolic markers using commercial PCR kits with cucurbit[8]urils (CB[8]) implemented to manipulate the activity of Taq DNA polymerase. CB[8] binds with the nonionic surfactants and displaces them from the polymerase surface, resulting in decreased enzyme activity. Meanwhile, the inhibited enzyme can be reversibly activated when spermine, the downstream metabolite of ornithine decarboxylase (ODC), is present in the sample, which competitively binds to CB[8] and recovers polymerase activity. CB[8] was implemented in conventional PCR kits not only to reduce false-positive results but also to extend the detection range of PCR technology. With this novel method to detect ODC in cell lysates containing both the nucleotides and intracellular metabolites, positive results were only observed in highly active HEK 293T cells, whereas silent cells treated with ODC inhibitor showed negative readouts, therefore providing a simple but elegant dual-modality PCR method for precision diagnosis.
研究领域:
分子生物学
·
生物技术
·
分析化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ENVIRONMENTAL POLLUTION
·
2019
·
IF 5.714
Pan-Pan Jia,Tai Sun,Muhammad Junaid,Li Yang,Yan-Bo Ma,Zhi-Song Cui,Da-Peng Wei,Hao-Fei Shi,De-Sheng Pei
Graphene family nanomaterials (GFNs) have attracted significant attention due to their unique characteristics and applications in the fields of biomedicine and nanotechnology. However, previous studies highlighted the in vitro and in vivo toxicity of GFNs with size and oxidation state differences are still elusive. Therefore, we prepared graphene (G) and graphene oxide (GO) of three different sizes (S-small, M-medium, and L-large), and characterized them using multiple surface-sensitive analytical techniques. In vitro assays using HEK 293T cells revealed that the small and large sizes of G and GO significantly reduced the cell viability and increased DNA damage, accompanying with activated reactive oxygen species (ROS) generation and induced various expressions of associated critical genetic markers. Moreover, the bacterial assays highlighted that G and GO caused strong acute toxicity on Tox2 bacteria. Effects of G were higher than GO and showed size dependent effect: L > M > S, while the medium size of GO induced mild genetic toxicity on RecA bacteria. In vivo assays revealed that exposure to G and GO caused the developmental toxicity, induced ROS generation, and activated related pathways (specifically GO) in zebrafish. Taken together, G showed stronger ability to decrease the survival rate and induce the acute toxicity, while GO showed obvious toxicity in terms of DNA damages, ROS generation, and abnormal gene expressions. Our findings highlighted that G and GO differentially induced toxicity based on their varying physical characteristics, especially sizes and oxidation state, and exposure concentrations and sensitivity of the employed in vitro and in vivo models. In short, this study provided deep insights on the negative effects of GFNs exposure.
研究领域:
纳米技术
·
生物医学
·
毒理学
·
细胞生物学
·
环境科学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
PEDIATRIC RESEARCH
·
2023
·
IF 3.6
Wang Qi,Shi Xulai,Li Ping-Ping,Gao Li,Zhou Yueyuan,Li Luyao,Ye Hao,Fu Xiaoqin,Li Peijun
Background Although previous studies show that microRNAs (miRNAs) can potentially be used as diagnostic markers for epilepsy, there are very few analyses of pediatric epilepsy patients. Methods miRNA profiles using miRNA-seq was performed on plasma samples from 14 pediatric epileptic patients and 14 healthy children. miRNA miR-27a-3p that were significantly changed between two groups were further evaluated. The potential target genes of miR-27a-3p were screened through unbiased mRNA-seq and further validated using Western blot and immunohistochemistry in HEK-293T cells and in the brains of mice with epilepsy induced by lithium chloride–pilocarpine. Results We found 82 upregulated and 76 downregulated miRNAs in the plasma from pediatric patients compared with controls ( p < 0.01), of which miR-27a-3p exhibited a very low p value ( p < 0.0001) and validated in additional plasma samples. Two genes, GOLM1 and LIMK1, whose mRNA levels were decreased ( p < 0.001) with the increase of miR-27a-3p were further validated in both HEK-293T cells and in epileptic mice. Conclusions MiR-27a-3p exhibits potential as a diagnostic and therapeutic marker for epilepsy. We postulate that additional studies on the downstream targets of miR-27a-3p will unravel its roles in epileptogenesis or disease progression. Impact
A total of 158 differentially expressed miRNAs were detected in plasma between epileptic and control children.
Plasma miR-27a-3p was one of the miRNAs with a low p value.
GOLM1 and LIMK1 were validated as downstream target genes of miR-27a-3p.
miR-27a-3p has potential as a diagnostic and therapeutic marker for epilepsy.
研究领域:
神经科学
·
分子生物学
·
儿科
·
生物标志物研究
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Small
·
2015
·Vol. 11
·期 47
·6338-6346
·
IF 8.368
Jing Tan,Chuanyu Sun,Ke Xu,Changchun Wang,Jia Guo
5-Aminolevulinic acid (ALA) is a widely used photodynamic therapy (PDT) prodrug in the clinic. It can be metalized to the photosensitizer PpIX, which produces toxic singlet oxygen to kill cancer cells upon visible light irradiation. Herein, a core/shell-structured vehicle is designed to comprise magnetite colloidal supraparticles (MCSPs) as cores and ALA-Zn II coordination polymers as shells (Fe 3 O 4 @ALA-Zn II ) for target pro-photosensitizer delivery. The coordination polymers with 2D layered structures are locally deposited on the MCSPs by the complexation of the ALA and Zn II ions, and are readily controlled by varying the feed precursors and reaction temperatures. The maximum conjugated ALA amount is up to 17%. The Fe 3 O 4 @ALA-Zn II microspheres exhibit pH-sensitive release of ALA in acidic environment and rapid magnetic responsiveness. Cytotoxicity results demonstrate that Fe 3 O 4 @ALA-Zn II shows a significant inhibitory effect to T24 cells and is nontoxic to 293T normal cells as exposed to the 630 nm visible light for a very short time, which may due to the selective accumulation of ALA-induced PpIX in T24 cancer cells. Compared to the ALA used alone, the coordination polymer form is more efficient because of the bioactivity of incorporated Zn ions despite underlying the same apoptosis mechanism as ALA agent.
研究领域:
材料科学
·
纳米技术
·
药学
·
肿瘤学
·
生物医学工程
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ChemNanoMat
·
2022
·Vol. 8
·期 3
·e202100453
·
IF 3.82
Qing Chang,Chenghao Liu,Zhiquan Xie,Qingfeng Shu,Yijun Xie,Qianqian Su,Xiaoyong Deng
Graphical An ultrasmall mesoporous silica delivery system (∼8 nm) with PEG and PEI dual-modification (FMSNs-PEG@PEI) was developed for loading negatively charged siRNA through electrostatic interaction. FMSNs-PEG@PEI can efficiently deliver siRNA into 293T cells and protect siRNA from being degraded. In addition, when GAPDH related gene was delivered by FMSNS-PEG@PEI, we observed a lower expression in GAPDH gene knockdown cells. This study offers new possibilities for advanced gene delivery research and gene therapy by using ultra-small mesoporous silica nanoparticles with minor cellular toxicity. Small interfering RNA (siRNA) for gene therapy has attracted great attention. However, due to easy degradation and inaccessibility to the cells, the gene silencing efficiency is not always satisfactory. To overcome this hurdle, developing an excellent nanocarrier to deliver siRNA into cancer cells is crucial. Fluorescent mesoporous silica nanoparticles are considered to be a superb nanocarrier for siRNA delivery systems because of its negligible toxicity and good biocompatibility. Herein, we developed an efficient fluorescent ultrasmall mesoporous silica nanocarrier for siRNA loading and delivery. The ultrasmall nanocarriers with polyethylenimine coating have a uniform size of about 10 nm. We find that the amino groups on polyethylenimine can protect siRNA from being degraded. Based on the fluorescence observation, we show that polyethylenimine-coated mesoporous silica nanocarrier loaded with siRNA can be uptake by 293T cells. Importantly, we also prove that siRNA can be delivered into 293T cells by our nanocarrier and subsequently inhibit the expression of related genes. The study provides insights into the development of nonviral fluorescence nanocarriers for siRNA delivery, which have important implications for targeted cancer therapy.
研究领域:
纳米技术
·
生物医学工程
·
分子生物学
·
基因治疗
✓ 已验证
产品关联
DOI
高影响因子
Biomaterials Research
·
2023
·Vol. 27
·期 1
·1-12
·
IF 11.3
Xu Xiaolin,Guan Wencai,Yu Xiaolei,Xu Guoxiong,Wang Chenglong
Background Protocell refers to the basic unit of life and synthetic molecular assembly with cell structure and function. The protocells have great applications in the field of biomedical technology. Simulating the morphology and function of cells is the key to the preparation of protocells. However, some organic solvents used in the preparation process of protocells would damage the function of the bioactive substance. Perfluorocarbon, which has no toxic effect on bioactive substances, is an ideal solvent for protocell preparation. However, perfluorocarbon cannot be emulsified with water because of its inertia. Methods Spheroids can be formed in nature even without emulsification, since liquid can reshape the morphology of the solid phase through the scouring action, even if there is no stable interface between the two phases. Inspired by the formation of natural spheroids such as pebbles, we developed non-interfacial self-assembly (NISA) of microdroplets as a step toward synthetic protocells, in which the inert perfluorocarbon was utilized to reshape the hydrogel through the scouring action. Results The synthetic protocells were successfully obtained by using NISA-based protocell techniques, with the morphology very similar to native cells. Then we simulated the cell transcription process in the synthetic protocell and used the protocell as an mRNA carrier to transfect 293T cells. The results showed that protocells delivered mRNAs, and successfully expressed proteins in 293T cells. Further, we used the NISA method to fabricate an artificial cell by extracting and reassembling the membrane, proteins, and genomes of ovarian cancer cells. The results showed that the recombination of tumor cells was successfully achieved with similar morphology as tumor cells. In addition, the synthetic protocell prepared by the NISA method was used to reverse cancer chemoresistance by restoring cellular calcium homeostasis, which verified the application value of the synthetic protocell as a drug carrier. Conclusion This synthetic protocell fabricated by the NISA method simulates the occurrence and development process of primitive life, which has great potential application value in mRNA vaccine, cancer immunotherapy, and drug delivery. Graphical Abstract
研究领域:
生物技术
·
细胞生物学
·
分子生物学
·
药学
·
癌症研究
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Journal of Pharmaceutical Analysis
·
2024
·
IF 6.1
Huanhuan Xue,Peijie Li,Jing Guo,Tinggui Chen,Shifei Li,Liwei Zhang
Ulcerative colitis (UC) is an idiopathic, relapsing, and etiologically complicated chronic inflammatory bowel disease. Despite substantial progress in the management of UC, the outcomes of mucosal barrier repair are unsatisfactory. In this study, phillygenin (PHI) treatment alleviated the symptoms of chronic colitis in mice, including body weight loss, severe disease activity index scores, colon shortening, splenomegaly, oxidative stress, and inflammatory response. In particular, PHI treatment ameliorated the tight junction proteins (TJs) reduction, fibrosis, apoptosis, and intestinal stem cell activity, indicating that PHI exerted beneficial effects on the intestinal mucosal barrier in mice with chronic colitis. In the NCM460 cells damage model, dextran sulfate sodium triggered the sequential induction of TJs reduction, fibrosis, and apoptosis. Takeda G protein-coupled receptor-5 (TGR5) dysfunction mediated NCM460 cell injury. Moreover, PHI treatment enhanced TJs and suppressed fibrosis and apoptosis to maintain NCM460 cell function, depending on TGR5 activation. PHI promoted TGR5 activation and elevated intracellular cyclic adenosine monophosphate levels in HEK 293T cells transfected with TGR5 expression plasmids. Cellular thermal shift assay and molecular docking studies confirmed that PHI directly binds to TGR5, indicating that PHI is an agonist of TGR5. The process of PERK-eIF2α pathway-mediated endoplasmic reticulum Ca 2+ release was involved in NCM460 cell injury as well, which was associated with TGR5 dysfunction. When NCM460 cells were pretreated with PHI, the PERK-eIF2α pathway and elevated Ca 2+ levels were blocked. In conclusion, our study demonstrated a novel mechanism that PHI inhibited the PERK-eIF2α-Ca 2+ pathway through TGR5 activation to against DSS-induced TJs reduction, fibrosis, and apoptosis.
研究领域:
胃肠病学
·
药理学
·
细胞生物学
·
炎症性肠病
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）