JOURNAL OF MOLECULAR LIQUIDS
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2023
·
IF 6
Yu wang,Liwen Zhuang,Yang wang,Chuanrun Li,Yichun Dong,Zhigang Lei,Yanyan Guo
The solubility and solvation mechanism of the Biopharmaceutical Classification System class II insoluble drug, ketoconazole (KCZ), were investigated in eight different types of imidazolium-based ionic liquids (ILs). An in vitro cytotoxicity test was conducted on Human immortalized keratinocytes cells to evaluate the potential toxicity of these ILs. The results showed no cytotoxicity for any of the imidazolium-based ILs tested. The solubility of KCZ was then examined in the eight imidazolium-based ILs at temperatures ranging from 298.15 K to 338.15 K. The findings revealed that the solubility of KCZ followed a specific order among the imidazolium-based ILs tested. The imidazolium-based IL [BMIM][Tf 2 N] exhibited the highest solubility of KCZ, followed by [HMIM][PF 6 ], [EMIM][Tf 2 N], [BMIM][TfO], [BMIM][PF 6 ], [EMIM][TfO], [BMIM][BF 4 ], and [EMIM][BF 4 ] in decreasing order. To understand the interaction mechanism between the imidazolium-based ILs and KCZ, the cations, anions and KCZ were analyzed using σ -profiles computed from both the COSMO-RS model and quantum chemical calculations. The results indicated that KCZ had strong hydrogen bond (HB) acceptors, particularly the oxygen atom of the carbonyl group and the nitrogen atom of the imidazole ring. These HB acceptors contributed to the formation of stronger HBs with the hydrogen atoms of cations compared to the oxygen atoms of the anions. The trend of HB strength followed [Emim] + > [Bmim] + > [Hmim] + > [Tf 2 N] − > [TfO] − > [PF 6 ] − > [BF 4 ] − . Based on these findings, it can be concluded that [BMIM][Tf 2 N], which has a strong HB donor cation and a strong HB acceptor anion, displayed the most effective dissolution of KCZ.
研究领域:
药学
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化学工程
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理论化学
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生物药剂学
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DOI
高影响因子
JOURNAL OF CELLULAR PHYSIOLOGY
·
2018
·Vol. 233
·期 12
·9763-9776
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IF 3.923
Xing Xie,Yuan Gan,Maoda Pang,Guoqing Shao,Lei Zhang,Beibei Liu,Qi Xu,Haiyan Wang,Yanyan Feng,Yanfei Yu,Rong Chen,Meng Wu,Zhenzhen Zhang,Lizhong Hua,Qiyan Xiong,Maojun Liu,Zhixin Feng
Primary porcine bronchial epithelial cells (PBECs) are an ideal model to study the molecular and pathogenic mechanisms of various porcine respiratory pathogens. However, the short lifespan of primary PBECs greatly limit their application. Here, we isolated and cultured primary PBECs and established immortalized PBECs by transfecting primary PBECs with the pEGFP-hTERT recombinant plasmid containing human telomerase reverse transcriptase (hTERT). Immortalized PBECs (hTERT-PBECs) retained the morphological and functional features of primary PBECs as indicated by cytokeratin 18 expression, telomerase activity assay, proliferation assays, karyotype analysis, and quantitative reverse-transcriptase polymerase chain reaction. Compared to primary PBECs, hTERT-PBECs had higher telomerase activity, extended replicative lifespan, and displayed enhanced proliferative activity. Moreover, this cell line is not transformed in vitro and does not exhibit a malignant phenotype in vivo, suggesting that it can be safely used in further studies. Besides, hTERT-PBECs were susceptible to swine influenza virus of H3N2 subtype and porcine circovirus type 2. In conclusion, the immortalized hTERT-PBECs represent a valuable in vitro model, which can be widely used in the study of porcine respiratory pathogenic infections.
研究领域:
细胞生物学
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病毒学
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分子生物学
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DOI
PubMed
高影响因子
Stem Cell Research & Therapy
·
2025
·Vol. 16
·期 1
·1-21
·
IF 7.3
Liao Guolong,Long Canling,Guo Rui,Wang Sushan,Han Ruijuan,Yang Jianhua,Liu Jia
Background Interstitial cystitis/bladder pain syndrome (IC/BPS) is a chronic condition characterized by debilitating pelvic pain. Mesenchymal stem cell-derived extracellular vesicles (MSC-EVs) are recognized as pivotal mediators of MSCs’ paracrine activity and represent a novel therapeutic approach for IC/BPS. However, their efficacy is hindered by the inherent variability of primary MSCs (pMSCs) from different donors and their susceptibility to senescence during culture expansion.Methods To overcome these challenges, we developed conditionally immortalized mesenchymal stem cells (iMSCs) using a doxycycline-regulated simian virus 40 Large T expression system in pMSCs. The conditioned proliferation capacity of iMSCs was evaluated using cell counting and immunocytochemistry. The expression of surface markers on iMSCs was assessed by flow cytometry. The osteogenic and adipogenic differentiation capabilities of iMSCs at different population doubling (PD) numbers were analyzed by qPCR, alizarin red staining, and oil red O staining. The EVs secreted by iMSCs were characterized using Western blot, scanning electron microscopy, and particle size analysis. In vitro and in vivo bladder inflammation models were used to evaluate the therapeutic effects of EVs on IC/BPS.Results These iMSCs exhibited precisely controlled proliferation, maintained surface marker expression and differentiation capacities, comparable to pMSCs up to PD 40. The characteristics of iMSC-EVs are equivalent to those of pMSCs. Furthermore, in vitro cellular experiments demonstrate that iMSC-EVs provide protective effects against LPS/ATP-induced damage in SV-HUC-1 cells. Additionally, administration of iMSC-EVs significantly enhanced tissue healing and anti-inflammatory capabilities in an IC/BPS animal model.Conclusion sIn summary, this approach produced a reliable source of functional MSCs and EVs, with iMSC-EVs demonstrating robust immunomodulatory properties and promoting tissue healing in IC/BPS. This method represents a promising alternative to pMSCs for IC/BPS therapy.
研究领域:
干细胞生物学
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泌尿学
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再生医学
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分子治疗
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细胞工程
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炎症研究
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产品关联
DOI
PubMed
高影响因子
ACTA HISTOCHEMICA
·
2020
·
IF 2.107
Xiaolong Sun,Xu Wang,Zhenyu Zhao,Jing Chen,Cheng Li,Gang Zhao
As one of the most frequent diabetic complications , diabetic foot ulcer (DFU) can cause limb ischemia or even amputation. Paeoniflorin (PF) has been reported to possess many kinds of biological functions, such as antioxidant and anti-inflammatory effects. However, the role of PF in DFU remains unknown. In this study, streptozotocin (STZ)-induced diabetic rat models and high glucose (HG)-treated Human immortalized keratinocytes (HaCaT) cells were established. Histological analysis, immunohistochemistry , Electrophoretic mobility shift assay , MTT assay , TUNEL assay , oxidative stress analysis, ELISA assay and western blot were used to investigate the role and underlying mechanisms of PF on healing in DFU. Our results showed that the STZ-induced diabetic rats had delayed wound healing compared with the normal rats, exhibited by intense oxidative DNA damage, low vascular endothelial growth factor (VEGF) and transforming growth factor β1 (TGF-β1) expression, as well as increased apoptosis . PF treatment activated the expression of nuclear factor-E2-related factor 2 (Nrf2) and improved wound healing in DFU rats. Our in vitro experiments confirmed that PF accelerated wound healing through the Nrf2 pathway under hyperglycemic conditions, with alleviated oxidative stress, increased cell proliferation and migration, decreased apoptosis, and increased the expression of VEGF and TGF-β1. Our study demonstrates the therapeutic benefits of PF in diabetic wound healing, which provides a reference for future clinical trials using PF in DFU treatment.
研究领域:
药理学
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糖尿病
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分子生物学
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细胞生物学
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伤口愈合
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DOI
PubMed
高影响因子
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
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2015
·Vol. 99
·期 21
·9135-9146
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IF 3.337
Xie Xing,Pang Maoda,Liang Shan,Yu Lei,Zhao Yanbing,Ma Ke,Kalhoro Dildar Hussain,Lu Chengping,Liu Yongjie
Dogs are susceptible to infectious diseases that occur primarily in the respiratory tract. The airway epithelium acts as a first line of defense and is constantly exposed to microorganisms present in the environment. Respiratory epithelial cells have recently gained wide use as a cell model for studying the pathogenesis of human, murine or swine respiratory pathogen infections. However, studies of the pathogenic mechanisms of canine pathogens have been hindered by the lack of reliable respiratory cell lines. Here, we cultured primary canine bronchiolar epithelial cells (CBECs), whose characteristics were confirmed by their expression of the epithelial cell-specific marker cytokeratin 18, and have provided protocols for their isolation and ex vivo expansion. Further, we established immortalized CBECs containing the human telomerase reverse transcriptase (hTERT) gene via transfection of primary CBECs with the recombinant plasmid pEGFP-hTERT. Immortalized bronchiolar epithelial cells (hTERT-CBECs) retain the morphological and functional features of primary CBECs, as indicated by reverse transcriptase polymerase chain reaction, proliferation assays, karyotype analysis, telomerase activity assay, and Western blotting, which demonstrate that hTERT-CBECs have higher telomerase activity, an extended proliferative lifespan, and a diploid complement of chromosomes, even after Passage 50. Moreover, this cell line is not transformed, as evaluated using soft agar assays and tumorigenicity analysis in nude mice, and can therefore be safely used in future studies. The isolation and establishment of stable hTERT-CBECs is of great importance for use as an in vitro model for mechanistic studies of canine pathogenic infections.
研究领域:
兽医学
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细胞生物学
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分子生物学
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微生物学
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DOI
PubMed
高影响因子
Parasites & Vectors
·
2024
·Vol. 17
·期 1
·1-17
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IF 3
Chen Huifang,Li Juan,Pan Xiaoting,Hu Zhichao,Cai Jianfeng,Xia Zijie,Qi Nanshan,Liao Shenquan,Spritzer Zachary,Bai Yinshan,Sun Mingfei
Background The gastrointestinal epithelium plays an important role in directing recognition by the immune system, and epithelial cells provide the host's front line of defense against microorganisms. However, it is difficult to cultivate avian intestinal epithelial cells in vitro for lengthy periods, and the lack of available cell lines limits the research on avian intestinal diseases and nutritional regulation. Chicken coccidiosis is a serious intestinal disease that causes significant economic losses in the poultry industry. In vitro, some cell line models are beneficial for the development of Eimeria species; however, only partial reproduction can be achieved. Therefore, we sought to develop a new model with both the natural host and epithelial cell phenotypes.Methods In this study, we use the SV40 large T antigen (SV40T) gene to generate an immortalized cell line. Single-cell screening technology was used to sort positive cell clusters with epithelial characteristics for passage. Polymerase chain reaction (PCR) identification, immunofluorescence detection, and bulk RNA sequencing analysis and validation were used to check the expression of epithelial cell markers and characterize the avian intestinal epithelial cell line (AIEC). AIECs were infected with sporozoites, and their ability to support the in vitro endogenous development of Eimeria tenella was assessed.Results This novel AIEC consistently expressed intestinal epithelial markers. Transcriptome assays revealed the upregulation of genes associated with proliferation and downregulation of genes associated with apoptosis. We sought to compare E. tenella infection between an existing fibroblast cell line (DF-1) and several passages of AIEC and found that the invasion efficiency was significantly increased relative to that of chicken fibroblast cell lines.Conclusion sAn AIEC will serve as a better in vitro research model, especially in the study of Eimeria species development and the mechanisms of parasite–host interactions. Using AIEC helps us understand the involvement of intestinal epithelial cells in the digestive tract and the immune defense of the chickens, which will contribute to the epithelial innate defense against microbial infection in the gastrointestinal tract.Graphical Abstract
研究领域:
兽医学
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细胞生物学
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分子生物学
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免疫学
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寄生虫学
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产品关联
DOI
PubMed
高影响因子
Biomaterials Advances
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2022
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IF 0
Shuwen Sun,Chuanbo Ding,Xinglong Liu,Yingchun Zhao,Jinping Zhang,Qiteng Ding,Yiwen Zhang,Yue Zhang,Mingqian Hao,Yinan Zheng,Wencong Liu,Min Yang
In modern clinical applications, wound healing remains a considerable challenge. Excessive inflammatory response is associated with delayed wound healing. In this study, we prepared composite nanofibrous membranes by mixing the Chinese herbal extract puerarin (PUE) with natural silk protein (SF) and synthetic polymer polyvinylpyrrolidone (PVP) using electrostatic spinning technique, and conducted a series of studies on the structural and biological properties of the fibrous membranes. The results showed that the loading of PUE increased the diameter, porosity and hydrophilicity of nanofibers, which were more favorable for cell adhesion and proliferation. ABTS radical scavenging assay also showed that the loading of PUE enhanced the antioxidant properties of the fibrous membranes. In addition, SF/PVP/PUE nanofibers are non-toxic and can be used as wound dressings. In vitro experiments showed that SF/PVP/PUE nanofibers could effectively alleviate lipopolysaccharide (LPS)-induced inflammation in Immortalized human keratinocytes (HaCaT) cells and down-regulate pro-inflammatory cytokine expression in cells. In vivo studies further showed that the SF/PVP/PUE nanofibers could effectively accelerate wound repair. The mechanism is that SF/PVP/PUE nanofibers can inhibit the activation and transduction of toll-like receptor 4/myeloid differentiation factor88/nuclear factor kappa B (TLR4/MyD88/NF-κB) and phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT) signaling pathways, thereby reducing the inflammatory response and achieving wound healing.
研究领域:
生物医学工程
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药学
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材料科学
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细胞生物学
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伤口愈合
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产品关联
DOI
PubMed
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
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2024
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IF 7.7
Haotian Deng,Xiong Zhang,Shuxian Su,Yuying Liu,Lishu Cui,Jianhao Zhao,Jianhua Rong
A novel poly(2-hydroxyethyl methacrylate- co -methacrylated hyaluronan- β -cyclodextrin) [p(HEMA- co -mHA- β -CD)] hydrogel was developed as a potential contact lens for ophthalmic disease. The hydrogel was synthesized from the copolymerization of 2-hydroxyethyl methacrylate (HEMA) monomer and mHA- β -CD as a hydrophilic macromolecular crosslinker. By adjusting the methacrylate substitution degree in hyaluronan (20–29 %) and the mHA- β -CD content (5–11 %), transparent p(HEMA-co-mHA- β -CD) hydrogels were achieved. p(HEMA- co -m 20 HA- β -CD) hydrogels exhibited an enhanced tensile modulus (from 0.35 to 0.88 MPa) with a decreased elongation at break (from 255 % to 108 %), meanwhile they showed increased hydrophilicity with a decreased water contact angle (from 83.4° to 48.6°) and an increased equilibrium water content (from 38.2 % to 46.4 %). Increasing the mHA- β -CD content resulted in a higher encapsulation and cumulative release of hydrophilic levofloxacin hydrochloride or hydrophobic puerarin, due to the improved hydrophilicity and the formation of β -CD/drug inclusion complexes. Compared with pHEMA hydrogel, p(HEMA-co-m 20 HA- β -CD) hydrogels better inhibited the deposition of lysozyme and bovine serum albumin, and the bacterial adhesion against S. aureus and E. coli . The hydrogels were stable at physiological conditions and non-toxic to immortalized human keratinocytes. With good mechanical properties, tear protein deposition resistance, antibacterial activity, and sustained drug delivery capabilities, p(HEMA-co-m 20 HA- β -CD) hydrogels were identified as a promising contact lens material for eye diseases.
研究领域:
材料科学
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生物医学工程
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药学
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眼科
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产品关联
DOI
PubMed
高影响因子
Frontiers in Microbiology
·
2021
·
IF 5.64
Xie Xing,Hao Fei,Chen Rong,Wang Jingjing,Wei Yanna,Liu Jin,Wang Haiyan,Zhang Zhenzhen,Bai Yun,Shao Guoqing,Xiong Qiyan,Feng Zhixin
Mycoplasma hyopneumoniae (Mhp) is the main pathogen that causes enzootic pneumonia, a disease that has a significant impact on the pig industry worldwide. The pathogenesis of enzootic pneumonia, especially possible virulence factors of Mhp, has still not been fully elucidated. The transcriptomic and proteomic analyses of different Mhp strains reported in the literature have revealed differences in virulence, and differences in RNA transcription levels between high- and low-virulence strains initially indicated that nicotinamide adenine dinucleotide (NADH)-dependent flavin oxidoreductase (NFOR) was related to Mhp pathogenicity. Prokaryotic expression and purification of the NFOR protein from Mhp were performed, a rabbit-derived polyclonal antibody against NFOR was prepared, and multiple sequence alignment and evolutionary analyses of Mhp NFOR were performed. For the first time, it was found that the NFOR protein was conserved among all Mhp strains, and NFOR was localized to the cell surface and could adhere to immortalized porcine bronchial epithelial cells (hTERT-PBECs). Adhesion to hTERT-PBECs could be specifically inhibited by an anti-NFOR polyclonal antibody, and the rates of adhesion to both high- and low-virulence strains, 168 and 168L, significantly decreased by more than 40%. Moreover, Mhp NFOR not only recognized and interacted with host fibronectin and plasminogen but also induced cellular oxidative stress and apoptosis in hTERT-PBECs. The release of lactate dehydrogenase by hTERT-PBECs incubated with Mhp NFOR was significantly positively correlated with the virulence of Mhp. Overall, in addition to being a metabolic enzyme related to oxidative stress, NFOR may also function as a potential novel virulence factor of Mhp, thus contributing to the pathogenesis of Mhp; these findings provide new ideas and theoretical support for studying the pathogenic mechanisms of other mycoplasmas.
研究领域:
微生物学
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病理学
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兽医学
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分子生物学
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细胞生物学
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产品关联
DOI
PubMed
高影响因子
CARBOHYDRATE POLYMERS
·
2022
·
IF 10.723
Yuhui Sheng,Xingchen Sun,Jingtian Han,Wei Hong,Jie Feng,Shuyang Xie,Youjie Li,Fangwei Yan,Keke Li,Baocheng Tian
An N -acetylcysteine functionalized chitosan oligosaccharide-palmitic acid conjugate (NAC-COS-PA) with bioadhesive and permeation promoting properties was synthesized to enhance transocular drug delivery. Flurbiprofen (FB) loaded self-assembled NAC-COS-PA nanomicelles (NAC-COS-PA-FB) were prepared and the drug loading was 7.35 ± 0.32%. Human immortalized corneal epithelial (HCE-T) cell cytotoxicity and hen's egg test-chorioallantoic membrane assays confirmed that the conjugate had good biocompatibility. The transportation efficiency of coumarin-6 (C6) loaded nanomicelles in the HCE-T cell monolayer was approximately 1.97 times higher than that of free C6. Decreased intracellular Ca 2+ concentration and cell membrane potential , increased cell membrane fluidity , and reversible changes in the F-actin cytoskeleton are presumed to be responsible for the enhanced drug permeation. NAC-COS-PA exhibited strong binding capacity with mucin and rabbit eyeball. In vivo pharmacokinetics indicated that the area under the curve (AUC 0 – 6 h ) and the maximum concentration (C max ) of NAC-COS-PA-FB were approximately 1.92 and 2.44 times that of the FB solution, respectively. NAC-COS-PA-FB demonstrated the best in vivo anti-inflammatory efficacy compared to unfunctionalized nanomicelles (COS-PA-FB) and FB solution. Consequently, NAC-COS-PA appears to be a promising bioadhesive carrier for ophthalmic delivery.
研究领域:
药学
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药物递送系统
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眼科学
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生物技术
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药代动力学
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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）