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Intracellular redox dyshomeostasis promoted by tumor microenvironment (TME) modulation has become an appealing therapeutic target for cancer management. Herein, a dual plasmonic Au/SF@Cu

Silk fibroin (SF), derived from Bombyx mori, is a category of fibrous protein with outstanding potential for applications in the biomedical and biotechnological fields. In spite of its many advantageous properties, the exploration of SF as a versatile nanodrug precursor for tumor therapy has still been restricted in recent years. Herein, a multifunctional SF-derived nanoplatform was facilely developed via encapsulating the photosensitizer chlorin e6 (Ce6) into MnO2-capped SF nanoparticles (NPs). SF@MnO2 nanocarriers were synthesized through a surface crystallization technique, using SF as a reductant and sacrificial template. Afterwards, Ce6 was covalently incorporated into the loose structure of the SF@MnO2 nanocarrier on the basis of adsorption to abundant peptide-binding sites. To modulate the tumor microenvironment (TME), SF@MnO2/Ce6 (SMC) NPs were capable of catalyzing the decomposition of H2O2 into O2, which can be converted into cytotoxic reactive oxygen species (ROS) during photodynamic therapy (PDT). Moreover, the MnO2 component was able to oxidize intracellular glutathione (GSH) into non-reducing glutathione disulfide (GSSG), and the consumption of GSH could significantly protect the local ROS from being reduced, which further augmented the therapeutic outcome of PDT. Via another angle, SMC NPs can produce strong hyperthermia under near-infrared (NIR) light activation, which was highly desirable for efficient photothermal therapy (PTT). Both in vitro and in vivo studies demonstrated the intense tumor inhibitory effects as a result of augmented PTT/PDT mediated by SMC NPs. We believe that this study may provide useful insights for employing SF-based nanocomposites for more medical applications in the near future.

BACKGROUND: Nowadays, biological tissue engineering is a growing field of research. Biocompatibility is a key indicator for measuring tissue engineering biomaterials, which is of great significance for the replacement and repair of damaged tissues. METHODS: In this study, using gelatin, carboxymethyl chitosan, and sodium alginate, a tissue engineering material scaffold that can carry cells was successfully prepared. The material was characterized by Fourier transforms infrared spectroscopy. In addition, the prepared scaffolds have physicochemical properties, such as swelling ratio, biodegradability. we observed the biocompatibility of the hydrogel to different adult stem cells (BMSCs and ADSCs) in vivo and in vitro. Adult stem cells were planted on gelatin-carboxymethyl chitosan-sodium alginate (Gel/SA/CMCS) hydrogels for 7 days in vitro, and the survival of stem cells in vitro was observed by live/died staining. Gel/SA/CMCS hydrogels loaded with stem cells were subcutaneously transplanted into nude mice for 14 days of in vivo culture observation. The survival of adult stem cells was observed by staining for stem cell surface markers (CD29, CD90) and Ki67. RESULTS: The scaffolds had a microporous structure with an appropriate pore size (about 80 μm). Live/died staining showed that adult stem cells could stably survive in Gel/SA/CMCS hydrogels for at least 7 days. After 14 days of culture in nude mice, Ki67 staining showed that the stem cells supported by Gel/SA/CMCS hydrogel still had high proliferation activity. CONCLUSION: Gel/SA/CMCSs hydrogel has a stable interpenetrating porous structure, suitable swelling performance and degradation rate, can promote and support the survival of adult stem cells in vivo and in vitro, and has good biocompatibility. Therefore, Gel/SA/CMCS hydrogel is a strong candidate for biological tissue engineering materials. GRAPHICAL ABSTRACT: [Image: see text]

Here, acidic tumor microenvironment (TME)-responsive nano-Bi

ObjectiveInfection is a major cause of death in patients with SLE. This study aimed to explore the infection rate in patients with SLE receiving a low dose of intravenous cyclophosphamide (IV-CYC).MethodsClinical parameters of 1022 patients with SLE from 24 hospitals in China were collected. Patients were divided into the short-interval and lower-dose (SILD, 400 mg every 2 weeks) IV-CYC group and the high-dose (HD, 500 mg/m2of body surface area every month) IV-CYC group. The clinical data and infection rate between the two groups were compared.ResultsCompared with HD IV-CYC, the infection rate of the SILD IV-CYC group was significantly lower (13.04% vs 22.27%, p=0.001). Respiratory tract infection (10.28% vs 15.23%, p=0.046) and skin/soft tissue infection (1.78% vs 4.3%, p=0.040) were significantly decreased in the SILD IV-CYC group. Moreover, infections occurred most likely in patients with SLE with leucopenia (OR 2.266, 95% CI 1.322 to 3.887, p=0.003), pulmonary arterial hypertension (OR 2.756, 95% CI 1.249 to 6.080, p=0.012) and >15 mg/day of glucocorticoid (OR 2.220, 95% CI 1.097 to 4.489, p=0.027).ConclusionsSILD IV-CYC showed a lower frequency of infection events than high-dose IV-CYC in patients with SLE.

Disturbance in redox homeostasis always leads to oxidative damages to cellular components, which inhibits cancer cell proliferation and causes tumor regression. Therefore, synergistic effects arising from cellular redox imbalance together with other treatment modalities are worth further investigation. Herein, a metal-organic framework nanosystem (NMOF) based on coordination between Fe (III) and 4,4,4,4-(porphine-5,10,15,20-tetrayl) tetrakis (benzoic acid) (TCPP) was synthesized through a one-pot method. After surface capping of silk fibroin (SF) to form NMOF@SF nanoparticles (NPs), this nanoplatform can serve as an eligible nanocarrier to deliver tirapazamine (TPZ), a hypoxia-activated precursor. As-developed NS@TPZ (NST) NPs remained inactive in the normal tissue, whereas became highly active upon endocytosis by tumor cells via glutathione (GSH)-mediated reduction of Fe (III) into Fe (II), further enabling Fe (II)-mediated chemodynamic therapy (CDT). Upon optical laser irradiation, TCPP-mediated photodynamic therapy (PDT) coordinated with CDT to aggravate intracellular oxidative stress. Thus, such reactive oxygen species accumulation and GSH deprivation contributed to a deleterious redox dyshomeostasis. On the other hand, local deoxygenation caused by PDT can increase the cytotoxicity of released TPZ, which significantly improved the integral therapeutic effectiveness relying on the combined redox balance disruption and bioreductive chemotherapy. More importantly, severe immunogenic cell death can be triggered by the combinatorial treatment modalities and the presence of SF, which facilitated an almost complete tumor eradication in vivo. Taken together, this paradigm provides an insightful strategy for tumor-specific redox dyshomeostasis treatment synergized by deoxygenation-driven chemotherapy, which can remarkably enhance antitumor efficacy with negligible adverse effects. Statement of Significance : Recently, silk fibroin (SF) has been demonstrated to be effective in activating antitumor immune system through polarization tumor-associated macrophages into M1 subtype. However, engineering SF into multifunctional nanocomposites is seldom reported for combination tumor therapy. In another aspect, disruption of redox homeostasis becomes increasingly attractive for tumor suppression with high clinical-relevance. Herein, we established a newfashioned NMOF nanosystem, named as NST, for tumor-specific redox dyshomeostasis treatment synergized by deoxygenation-driven chemotherapy. This platform takes advantages of Fe2+/Fe3+ coupled Fenton-like reaction and GSH depletion, as well as TCPP-mediated photosensitization for admirable redox unbalancing, which further initiates hypoxia-relevant toxin of TPZ for chemotherapy. Finally, combinatorial treatments and the presence of SF could trigger ICD for rendering a complete tumor eradication in vivo.

The intracellular PI3K, MAPK, and mTOR network controls morphogenesis by regulating the distribution of cell surface receptors., The patterning of epithelial buds is determined by the underlying signaling network. Here, we study the cross-talk between phosphoinositide 3-kinase (PI3K) and Ras signaling during lacrimal gland budding morphogenesis. Our results show that PI3K is activated by both the p85-mediated insulin-like growth factor (IGF) and Ras-mediated fibroblast growth factor (FGF) signaling. On the other hand, PI3K also promotes extracellular signal–regulated kinase (ERK) signaling via a direct interaction with Ras. Both PI3K and ERK are upstream regulators of mammalian target of rapamycin (mTOR), and, together, they prevent expansion of epidermal growth factor (EGF) receptor expression from the lacrimal gland stalk to the bud region. We further show that this suppression of EGF signaling is necessary for induction of lacrimal gland buds. These results reveal that the interplay between PI3K, mitogen-activated protein kinase, and mTOR mediates the cross-talk among FGF, IGF, and EGF signaling in support of lacrimal gland development.

The therapeutic exploration of nano-zirconia semiconductor largely remains untouched in the field of fundamental science to date. Here, a robust nano-sonosensitizer of ZrO

Silk fibroin (SF), derived from Bombyx mori, is a category of fibrous protein with outstanding potential for applications in the biomedical and biotechnological fields. In spite of its many advantageous properties, the exploration of SF as a versatile nanodrug precursor for tumor therapy has still been restricted in recent years. Herein, a multifunctional SF-derived nanoplatform was facilely developed via encapsulating the photosensitizer chlorin e6 (Ce6) into MnO2-capped SF nanoparticles (NPs). SF@MnO2 nanocarriers were synthesized through a surface crystallization technique, using SF as a reductant and sacrificial template. Afterwards, Ce6 was covalently incorporated into the loose structure of the SF@MnO2 nanocarrier on the basis of adsorption to abundant peptide-binding sites. To modulate the tumor microenvironment (TME), SF@MnO2/Ce6 (SMC) NPs were capable of catalyzing the decomposition of H2O2 into O2, which can be converted into cytotoxic reactive oxygen species (ROS) during photodynamic therapy (PDT). Moreover, the MnO2 component was able to oxidize intracellular glutathione (GSH) into non-reducing glutathione disulfide (GSSG), and the consumption of GSH could significantly protect the local ROS from being reduced, which further augmented the therapeutic outcome of PDT. Via another angle, SMC NPs can produce strong hyperthermia under near-infrared (NIR) light activation, which was highly desirable for efficient photothermal therapy (PTT). Both in vitro and in vivo studies demonstrated the intense tumor inhibitory effects as a result of augmented PTT/PDT mediated by SMC NPs. We believe that this study may provide useful insights for employing SF-based nanocomposites for more medical applications in the near future.

Human endogenous retrovirus (HERVs), originating from exogenous retroviral infections of germ cells millions of years ago, have the potential for human diseases. Syncytin-1, an envelope protein encoded by the HERV W family, participates in the contexts of schizophrenia, multiple sclerosis, diabetes, and several types of cancers. Nevertheless, there is no report on the expression pattern and potential mechanism of Syncytin-1 in HCC. Here we found Syncytin-1 expression was up-regulated in HCC compared to adjacent non-tumorous tissues, especially in advanced HCC. Syncytin-1 was an independent risk factor to predict vascular invasion, metastasis, larger tumor size, and poor prognosis in HCC patients. Further analysis discovered that Syncytin-1 overexpression positively associated with HCC patients with serum HBsAg positive. Functional experiments in vitro and in vivo demonstrated that Syncytin-1 enhanced cell proliferation, metastasis, and tumorigenicity in HCC. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis suggested that the mitogen-activated protein kinase (MEK)/extracellular signal-regulated protein kinase (ERK) pathway was involved in HCC. Our clinical data indicated that the levels of phosphorylation MEK1/2 and ERK1/2 were increased in HCC comparing with adjacent non-tumorous tissues. It showed the linear correlation between Syncytin-1 expression and upregulated MEK1/2 and ERK1/2 phosphorylation levels in HCC. Furthermore, Syncytin-1 activated MEK/ERK pathway in HCC cells. In-depth research showed that the inflammation-activated MEK/ERK pathway was essential in Syncytin-1 promoted hepatocarcinogenesis. Syncytin-1 suppressed doxorubicin-induced apoptosis via MEK/ERK cascade. In conclusion, Syncytin-1 promoted HCC progression and doxorubicin resistance via the inflammation-activated MEK/ERK pathway. Our findings revealed that Syncytin-1 was a potential prognostic biomarker and therapeutic target for HCC.

In this meta-analysis study, we compared the oncological and functional outcomes of intrafascial radical prostatectomy (IFRP) with non-intrafascial radical prostatectomy (NIFRP) in the treatment of patients with low risk localized prostate cancer (PCa). Relevant articles were identified by searching PubMed, EMBASE, Cochrane Library, Ovid, and the ISI Web of Knowledge databases. A total of 2096 patients were included from 7 eligible studies. Results of the pooled data showed that the oncological outcomes including gleason score, positive surgical margin and biochemical free survival rates were similar between the two groups. IFRP was superior to NIFRP with lower postoperative complication rates (RR 0.57, 95% CI 0.38, 0.85, p = 0.006), higher continence rates at 3 months post-operation (RR: 1.14; 95% CI, 1.04, 1.26; p = 0.006), and higher potency rates at 6 months (RR: 1.53; 95% CI, 1.07, 2.18; p = 0.02) and 12 months post-operation (RR: 1.38; 95% CI, 1.11, 1.73; p = 0.005). Additionally, there was a tendency towards higher potency rate in patients ≤65 years old compared with patients >65 years old after IFRP. Overall, these findings suggest that IFRP in young patients with low risk localized PCa had less postoperative complications, shortened time to return to continence and improved potency rate without compromising complete tumor control.

Here, acidic tumor microenvironment (TME)-responsive nano-Bi2 Se3 @MnCaP, as a near-infrared-II (NIR-II) biowindow-triggered free radical generator for hypoxia-irrelevant phototherapy, is elaborately developed by biomimetic mineralization of MnCaP onto 2, 2'-azobis[2-(2-imidazolin-2-yl)propane] dihydrochloride (AIPH)-loaded mesoporous nano-Bi2 Se3 to form Bi2 Se3 /AIPH@MnCaP (BAM). Surface mineral of MnCaP can be degraded under mild acidity, leading to the release of both Mn2+ and AIPH. The leached Mn2+ not only facilitates chemodynamic therapy (CDT) via hydroxyl radicals (• OH) from Mn2+ -mediated Fenton-like reaction but also acts as contrast agent for magnetic resonance imaging. In another aspect, the splendid photothermal conversion capacity of BAM enables a rapid hyperthermia generation under NIR-II laser irradiation for photothermal therapy (PTT). Simultaneously, the local thermal shock can induce the disintegration of AIPH to generate alkyl radicals (• R) for thermodynamic therapy (TDT) and accelerate Fenton-like reaction rate to augment CDT efficacy. The strong synergistic effects from cooperative CDT/PTT/TDT are applied to 4T1 tumor suppression with minimal side effects. Importantly, the combination therapy can effectively trigger immunogenetic cell death and enhance antitumor immunity for systemic tumor eradication. Collectively, this proof-of-concept study demonstrates a more efficacious and safer strategy for oxygenation-independent phototherapy, which holds a good potential for clinical translation in cancer management.

Background Rheumatic disease is caused by immune abnormality,which results in the immune response of the body to autoantigens and causes tissue damage.The immune system eliminates or inhibits the growth of tumor cells through various ways.When the immune function abnormality leads to the escape of immune surveillance of tumor cells,it can lead to tumorigenesis.The risk of malignant tumors is higher than ordinary people in systemic lupus erythematosus (SLE) patients, which were mainly hematological tumors,especially non-Hodgkin lymphoma.The risk of Hodgkin lymphoma, leukemia and multiple myeloma was also increased, which was associated with BAFF changes,APRIL signal transduction, increased IL-6 activity and EB virus infection.Increased risk of malignant tumors in SLE patients,including lung cancer,hepatobiliary cancer,thyroid cancer, vulvar/vaginal malignancies and cervical dysplasia,was also reported,which was associated with SLE disease activity in specific organs,inflammatory stimulation,oxidative damage,changes in viral clearance and/or specific drug use. Methods Twenty SLE patients with malignant tumors admitted to the General Hospital of Ningxia Medical University from October 2008 to October 2018 were selected.Their sex,age,time of diagnosis of rheumatism and tumors, types of tumors, clinical stages and prognosis were summarized and analyzed. Results Malignant tumors occurred in 20 (0.50%) of 3970 SLE patients,the average age was 54.43±7.95.There were 4 cases of lung cancer,3 cases of non-Hodgkin lymphoma and liver cancer,2 cases of thyroid cancer,uterine cancer and skin cancer,Multiple myeloma, gastric cancer, pancreatic cancer and nasopharyngeal carcinoma were found in 1 case respectively.The median course of malignant tumors in SLE patients was 3.42 years.79.41%of the patients were diagnosed before tumors,16.35% of the cases occurred at the same time,And only 4.24% of tumors were diagnosed before SLE.Phase IV was the most common clinical stage in the diagnosis of tumors,accounting for 59.74%.In SLE patients with tumors, the survival time ranged from 1 month to 39 months, with a median survival time of 9.2 months. Conclusions Most malignant tumors are found after SLE diagnosis, with a median course of 3.42 years,and occur more frequently between the ages of 50 and 60.SLE may be associated with various types of malignant tumors, Lung cancer, non-Hodgkin’s lymphoma and Liver cancer were the most common tumors,and the clinical staging of tumors is late and the prognosis is poor,with the median survival time is 9.2 months. Funding Source(s): None

How multiple receptor tyrosine kinases coordinate cell fate determination is yet to be elucidated. We show here that the receptor for platelet-derived growth factor (PDGF) signaling recruits the p85 subunit of Phosphoinositide 3-kinase (PI3K) to regulate mammalian lens development. Activation of PI3K signaling not only prevents B-cell lymphoma 2 (BCL2)-Associated X (Bax)- and BCL2 Antagonist/Killer (Bak)-mediated apoptosis but also promotes Notch signaling to prevent premature cell differentiation. Reducing PI3K activity destabilizes the Notch intracellular domain, while the constitutive activation of Notch reverses the PI3K deficiency phenotype. In contrast, fibroblast growth factor receptors (FGFRs) recruit Fibroblast Growth Factor Receptor Substrate 2 (Frs2) and Rous sarcoma oncogene (Src) Homology Phosphatase 2 (Shp2) to activate Mitogen-Activated Protein Kinase (MAPK) signaling, which induces the Notch ligand Jagged 1 (Jag1) and promotes cell differentiation. Inactivation of Shp2 restored the proper timing of differentiation in the p85 mutant lens, demonstrating the antagonistic interaction between FGF-induced MAPK and PDGF-induced PI3K signaling. By selective activation of PI3K and MAPK, PDGF and FGF cooperate with and oppose each other to balance progenitor cell maintenance and differentiation., Author summary A central aim in understanding cell signaling is to decode the cellular logic that underlies the functional specificity of growth factors. Although these factors are known to activate a common set of intracellular pathways, they nevertheless play specific roles in development and physiology. Using lens development in mice as a model, we show that fibroblast growth factor (FGF) and platelet-derived growth factor (PDGF) antagonize each other through their intrinsic biases toward distinct downstream targets. While FGF primarily induces the Ras–Mitogen-Activated Protein Kinase (MAPK) axis to promote lens cell differentiation, PDGF preferentially stimulates Phosphoinositide 3-kinase (PI3K) to enhance Notch signaling, which is necessary for maintaining the lens progenitor cell pool. By revealing the intricate interactions between PDGF, FGF, and Notch, we present a paradigm for how signaling crosstalk enables balanced growth and differentiation in multicellular organisms.

Neat crosslinked polymeric ionic liquid (PIL) sorbent coatings for solid-phase microextraction (SPME) compatible with high-performance liquid chromatography (HPLC) are reported for the first time. Six structurally different PILs were crosslinked to nitinol supports and applied for the determination of select pharmaceutical drugs, phenolics, and insecticides. Sampling conditions including sample solution pH, extraction time, desorption solvent, desorption time, and desorption solvent volume were optimized using design of experiment (DOE). The developed PIL sorbent coatings were stable when performing extractions under acidic pH and remained intact in various organic desorption solvents (i.e., methanol, acetonitrile, acetone). The PIL-based sorbent coating polymerized from the IL monomer 1-vinyl-3-(10-hydroxydecyl) imidazolium chloride [VC10OHIM][Cl] and IL crosslinker 1,12-di(3-vinylbenzylimidazolium) dodecane dichloride [(VBIM)2C12] 2[Cl] exhibited superior extraction performance compared to the other studied PILs. The extraction efficiency of pharmaceutical drugs and phenolics increased when the film thickness of the PIL-based sorbent coating was increased while many insecticides were largely unaffected. Satisfactory analytical performance was obtained with limits of detection (LODs) ranging from 0.2 to 2 μg L(-1) for the target analytes. The accuracy of the analytical method was examined by studying the relative recovery of analytes in real water samples, including tap water and lake water, with recoveries varying from 50.2% to 115.9% and from 48.8% to 116.6%, respectively.

The use of highly robust polymeric ionic liquid (PIL) sorbent coatings combined with direct immersion (DI) and headspace (HS) modes in a single solid-phase microextraction (SPME) procedure coupled to gas chromatography (GC) is reported. Target analytes with a wide range of volatilities including polycyclic aromatic hydrocarbons, organochlorine pesticides, and phthalate esters were studied. Three PIL-based sorbent coatings were evaluated and the highest extraction efficiency for most analytes was obtained using the PIL coating consisting of the 1-vinylbenzyl-3-hexadecylimidazolium bis[(trifluoromethyl)sulfonyl]imide [VBC16IM][NTf2] IL monomer and the [(VBIM)2C12]2[NTf2] IL crosslinker. The combined DI-HS mode allowed for a compromise extraction condition among the different classes of analytes compared to the use of DI and HS modes separately. An extraction temperature of 40 °C and an extraction time of 50 min with 50% of this time in HS mode was optimized by application of a central composite design. Satisfactory analytical performance was achieved with limits of detection varying from 0.003 to 0.15 μg L−1 and calibration curve correlation coefficients ranging from 0.980 to 0.999. Analyte relative recoveries from tap and lake water ranged from 52.3 to 116.8% with relative standard deviations ranging from 0.3 to 19.3%.

Objective To assess the possibility of ureter tissue engineering using vessel extracellular matrix (VECM) and differentiated urine-derived stem cells (USCs) in a rabbit model. Methods VECM was prepared by a modified technique. USCs were isolated from human urine samples and cultured with an induction medium for the differentiation of the cells into urothelium and smooth muscle phenotypes. For contractile phenotype conversion, the induced smooth muscle cells were transfected with the miR-199a-5p plasmid. The differentiated cells were seeded onto VECM and cultured under dynamic conditions in vitro for 2 weeks. The graft was tubularized and wrapped by two layers of the omentum of a rabbit for vascularization. Then, the maturated graft was used for ureter reconstruction in vivo. Results VECM has microporous structures that allow cell infiltration and exhibit adequate biocompatibility with seeding cells. USCs were isolated and identified by flow cytometry. After induction, the urothelium phenotype gene was confirmed at mRNA and protein levels. With the combined induction by TGF-β1 and miR-199a-5p, the differentiated cells can express the smooth muscle phenotype gene and convert to the contractile phenotype. After seeding cells onto VECM, the induced urothelium cells formed a single epithelial layer, and the induced smooth muscle cells formed a few cell layers during dynamic culture. After 3 weeks of omental maturation, tubular graft was vascularized. At 2 months post ureter reconstruction, histological evaluation showed a clearly layered structure of ureter with multilayered urothelium over the organized smooth muscle tissue. Conclusion By seeding differentiated USCs onto VECM, a tissue-engineered graft could form multilayered urothelium and organized smooth muscle tissue after ureteral reconstruction in vivo. Statement of significance Cell-based tissue engineering offers an alternative technique for urinary tract reconstruction. In this work, we describe a novel strategy for ureter tissue engineering. We modified the techniques of vessel extracellular matrix (VECM) preparation and used a dynamic culture system for seeding cells onto VECM. We found that VECM had the trait of containing VEGF and exhibited blood vessel formation potential. Urine-derived stem cells (USCs) could be differentiated into urothelial cells and functional contractile phenotype smooth muscle cells in vitro. By seeding differentiated USCs onto VECM, a tissue-engineered graft could form multilayered urothelium and organized smooth muscle tissue after ureteral reconstruction in vivo. This strategy might be applied in clinical research for the treatment of long-segment ureteral defect.

DNA extraction represents a significant bottleneck in nucleic acid analysis. In this study, hydrophobic magnetic ionic liquids (MILs) were synthesized and employed as solvents for the rapid and efficient extraction of DNA from aqueous solution. The DNA-enriched microdroplets were manipulated by application of a magnetic field. The three MILs examined in this study exhibited unique DNA extraction capabilities when applied toward a variety of DNA samples and matrices. High extraction efficiencies were obtained for smaller single-stranded and double-stranded DNA using the benzyltrioctylammonium bromotrichloroferrate(III) ([(C8)3BnN(+)][FeCl3Br(-)]) MIL, while the dicationic 1,12-di(3-hexadecylbenzimidazolium)dodecane bis[(trifluoromethyl)sulfonyl]imide bromotrichloroferrate(III) ([(C16BnIM)2C12(2+)][NTf2(-), FeCl3Br(-)]) MIL produced higher extraction efficiencies for larger DNA molecules. The MIL-based method was also employed for the extraction of DNA from a complex matrix containing albumin, revealing a competitive extraction behavior for the trihexyl(tetradecyl)phosphonium tetrachloroferrate(III) ([P6,6,6,14(+)][FeCl4(-)]) MIL in contrast to the [(C8)3BnN(+)][FeCl3Br(-)] MIL, which resulted in significantly less coextraction of albumin. The MIL-DNA method was employed for the extraction of plasmid DNA from bacterial cell lysate. DNA of sufficient quality and quantity for polymerase chain reaction (PCR) amplification was recovered from the MIL extraction phase, demonstrating the feasibility of MIL-based DNA sample preparation prior to downstream analysis.

Magnetic ionic liquids (MILs) are a subclass of ionic liquids (ILs) containing paramagnetic components and are readily manipulated by an external magnetic field. Due to their hydrophilic nature, very few applications of MILs in aqueous systems have been reported. In this study, three general classes of hydrophobic MILs including monocationic, symmetrical/unsymmetrical dicationic, and symmetrical/unsymmetrical tricationic MILs were synthesized and characterized. By tuning the structure of the MIL, various physicochemical properties including water solubility, magnetic susceptibility, and melting point were regulated. MILs synthesized with the benzimidazolium cation were shown to exhibit lower water solubility (0.1% (w/v)) when compared to those containing imidazolium cations (0.25% (w/v)). By incorporating asymmetry into the cationic component of the MIL, the melting point of dicationic MILs was lowered while the effective magnetic moment (μeff) and hydrophobicity remained unchanged. Tricationic MILs paire...

The extraction performance of four polymeric ionic liquid (PIL)-based solid-phase microextraction (SPME) coatings has been studied and compared to that of commercial SPME coatings for the extraction of 16 volatile compounds in cheeses. The analytes include 2 free fatty acids, 2 aldehydes, 2 ketones and 10 phenols and were determined by headspace (HS)-SPME coupled to gas chromatography (GC) with flame-ionization detection (FID). The PIL-based coatings produced by UV co-polymerization were more efficient than PIL-based coatings produced by thermal AIBN polymerization. Partition coefficients of analytes between the sample and the coating (Kfs) were estimated for all PIL-based coatings and the commercial SPME fiber showing the best performance among the commercial fibers tested: carboxen-polydimethylsyloxane (CAR-PDMS). For the PIL-based fibers, the highest K(fs) value (1.96 ± 0.03) was obtained for eugenol. The normalized calibration slope, which takes into account the SPME coating thickness, was also used as a simpler approximate tool to compare the nature of the coating within the determinations, with results entirely comparable to those obtained with estimated K(fs) values. The PIL-based materials obtained by UV co-polymerization containing the 1-vinyl-3-hexylimidazolium chloride IL monomer and 1,12-di(3-vinylimiazolium)dodecane dibromide IL crosslinker exhibited the best performance in the extraction of the select analytes from cheeses. Despite a coating thickness of only 7 µm, this copolymeric sorbent coating was capable of quantitating analytes in HS-SPME in a 30 to 2000 µg L(-1) concentration range, with correlation coefficient (R) values higher than 0.9938, inter-day precision values (as relative standard deviation in %) varying from 6.1 to 20%, and detection limits down to 1.6 µg L(-1).

Urolithins were the metabolites of ellagic acid by intestinal flora in gastrointestinal tract. In previous research, it was found that urolithins could mainly inhibit prostate cancer and colon cancer cell growth. However, there is no report about bladder cancer therapy of urolithins. In this paper, three urolithin-type compounds (urolithin A, urolithin B, 8-OMe-urolithin A) and ellagic acid were evaluated for antiproliferative activity in vitro against human bladder cancer cell lines T24. The IC₅₀ values for T24 cell inhibition were 43.9, 35.2, 46.3 and 33.7 μM for urolithin A, urolithin B, 8-OMe-urolithin A and ellagic acid, respectively. After the administration of urolithins and ellagic acid, we found these compounds could increase mRNA and protein expression of Phospho-p38 MAPK, and decrease mRNA and protein expression of MEKK1 and Phospho-c-Jun in T24 cells. Caspase-3 was also activated and PPAR-γ protein expression increased in drug-induced apoptosis. And what's more, the antioxidant assay afforded by three urolithins and EA treatments were associated with decreases in the intracellular ROS and MDA levels, and increased SOD activity in H₂O₂-treated T24 cells. The results suggested that these compounds could inhibit cell proliferation by p38-MAPK and/or c-Jun medicated caspase-3 activation and reduce the oxidative stress status in bladder cancer.

Caffeine and aspirin have been suggested to be involved in neurologic diseases, such as schizophrenia, and previous data have revealed that abnormal expression of HERV-W elements may be an important factor in the etiopathogenesis of those diseases. In this article, we reported that caffeine and aspirin contributed to the expression of HERV-W env and gag in Human SH-SY5Y neuroblastoma cells. Semi-quantitative RT-PCR and quantitative Real-time PCR were used to detect the mRNA of HERV-W env and gag in cells exposed to caffeine or aspirin. Western blotting was used to detect the protein of HERV-W env. Luciferase activity assay was employed to detect the activity of HERV-W env promoter. It was found that both caffeine and aspirin could increase the expression of HERV-W env and gag in human SH-SY5Y neuroblastoma cells. Caffeine could activate the HERV-W env promoter, while aspirin could not. With previous studies we can conjecture that HERVs might play a bridging role between environmental factors, such as drugs and neurologic diseases.

Sample preparation is often considered to be a bottleneck in most analytical methods. Solid-phase microextraction (SPME) consolidates sampling and sample preparation into one step. The unique properties of ionic liquids (ILs) and polymeric ILs (PILs) have been exploited as sorbent coatings in SPME. Due to their tunable structures, these materials exhibit unique selectivity for various analytes. This article discusses the overall progression of IL and PIL-based sorbent coatings in SPME. The structural engineering and design of selective sorbent coatings is discussed to demonstrate the versatility of these materials in numerous applications.

Ionic liquids (ILs) are an attractive class of non-molecular solvents whose versatility allows them to be incorporated into various chromatographic and microextraction methods.

Polysaccharides extracted from Lycium barbarum exhibit antioxidant properties. We hypothesized that these polysaccharides resist oxidative stress-induced neuronal damage following cavernous nerve injury. In this study, rat models were intragastrically administered Lycium barbarum polysaccharides for 2 weeks at 1, 7, and 14 days after cavernous nerve injury. Serum superoxide dismutase and glutathione peroxidase activities significantly increased at 1 and 2 weeks post-injury. Serum malondialdehyde levels decreased at 2 and 4 weeks. At 12 weeks, peak intracavernous pressure, the number of myelinated axons and nicotinamide adenine dinucleotide phosphate-diaphorase-positive nerve fibers, levels of phospho-endothelial nitric oxide synthase protein and 3-nitrotyrosine were higher in rats administered at 1 day post-injury compared with rats administered at 7 and 14 days post-injury. These findings suggest that application of Lycium barbarum polysaccharides following cavernous nerve crush injury effectively promotes nerve regeneration and erectile functional recovery. This neuroregenerative effect was most effective in rats orally administered Lycium barbarum polysaccharides at 1 day after cavernous nerve crush injury.

Three hydrophobic magnetic ionic liquids (MILs) containing the tetrachloromanganate(II) (MnCl4(2-)) anion, namely, aliquat tetrachloromanganate(II) ([Aliquat(+)]2[MnCl4(2-)]), methyltrioctylammonium [MnCl4(2-)] ([N1,8,8,8(+)]2[MnCl4(2-)]), and trihexyltetradecylphosphonium [MnCl4(2-)] ([P6,6,6,14(+)]2[MnCl4(2-)]) were employed as extraction solvents in DLLME coupled to high-performance liquid chromatography (HPLC) employing UV detection. The MILs were developed with the features of magnetic susceptibility to permit rapid retrieval of the extraction solvent, hydrophobicity to allow for phase separation from water, and mobile phase compatibility with reversed phase HPLC. Additionally, the MILs were customized to minimize hydrolysis of the anionic component in aqueous media as well as reduce absorbance when subjected to HPLC. The three MILs were applied for the extraction of pharmaceutical drugs, phenolics, insecticides, and polycyclic aromatic hydrocarbons. The disperser solvent type, disperser solvent volume, mass of MIL, extraction time, the pH of the sample solution, and salt concentration were studied in order to achieve optimal extraction efficiency for each MIL. The [P6,6,6,14(+)]2[MnCl4(2-)] MIL exhibited the best extraction efficiencies for most of the target analytes compared to the other MILs. Good linearity was obtained using this MIL with correlation coefficients (R) varying from 0.997 to 0.999. The limits of detection (LODs) of all analytes ranged from 0.25 to 1.00μgL(-1). The relative recovery was studied in lake water and river water. The relative recovery in lake water varied from 53.8% to 114.7% at a spiked concentration of 20μgL(-1) (5μgL(-1) for phenanthrene) and from 52.1% to 106.7% at 150μgL(-1) (37.5μgL(-1) for phenanthrene). In river water, the relative recovery varied from 44.6% to 110.7% at a spiked concentration of 20μgL(-1) (5μgL(-1) for phenanthrene) and 42.9% to 83.6% at 150μgL(-1) (37.5μgL(-1) for phenanthrene).

Objective This study aimed to investigate the effect of Lycium barbarum polysaccharides (LBP) on erectile function recovery in a rat model of cavernous nerve (CN) crush injury. Methods Rats were randomly divided into five groups: sham group, injured control group, and three experimental groups (group A, B, C, with an oral administration of LBP at 1, 7, and 14 day respectively after CN crush injury for 2 weeks). Malondialdehyde (MDA), super oxide dismutase (SOD) and glutathione peroxidase (GPX) were evaluated at 1, 2, 4, and 12 week. Intracavernous pressure (ICP) and mean arterial pressure (MAP) were assessed at 12 week. Myelinated axons of CN and Nitric Oxide Synthase (NOS) containing nerve fibres were evaluated. Endothelial NOS (eNOS) protein expression and cyclic guanosine monophosphate (cGMP) level were measured. Results SOD and GPX activities in experimental group A were significantly higher than those in other four groups at 1 and 2 week. MDA levels in experimental group A were significantly lower than those in other four groups at 2 and 4 week. The peak ICP and peak ICP/MAP ratio in experimental group A were significantly higher than those in injured control group and other two experimental groups at 12 week. The number of myelinated axons, NADPH-diaphorase-positive nerve fibres, phospho-eNOS protein expression and cGMP level were in concordance with the results of peak ICP and peak ICP/MAP ratio. Conclusions The application of LBP in the first 2 weeks after CN crush injury promotes nerve regeneration and erectile function recovery via an antioxidative mechanism.

Background The purpose of the present study was to assess the differentiation potential of adipose-derived stem cells (ASCs) into smooth muscle cells (SMCs) and their potential for promoting regeneration of smooth muscle for ureteral tissue engineering. Methods ASCs were isolated, proliferated, and identified in vitro. SMC differentiation was induced using SMC induction medium. Gene expression was evaluated by quantitative polymerase chain reaction, immunofluorescence, and Western blotting. Vessel extracellular matrix was obtained by a decellularization process. The induced cells were seeded onto vessel extracellular matrix for ureter reconstitution. Grafts were obtained for evolutionary histologic studies. Renal function and ureteral patency was evaluated by intravenous urography at 16 wk. Results Flow cytometry demonstrated that the ASCs expressed CD90, but did not express CD45 or CD34. After 6 wk of induction, upregulation of α-smooth muscle actin expression was determined by quantitative polymerase chain reaction, and smooth muscle myosin heavy chain expression was confirmed by immunofluorescence and Western blotting in the induced cells. Vessel extracellular matrix exhibited a nontoxic and bioactive effect on the induced cells. Histologically, stratified urothelium and organized muscle bundles were observed in the grafts at 16 wk. Intravenous urography demonstrated no ureteral stricture or hydroureteronephrosis. Conclusions These results have demonstrated that ASCs can be differentiated into SMCs and this potential promoted smooth muscle regeneration for ureteral tissue engineering.

A high-throughput method for the production of solid-phase microextraction (SPME) sorbent coatings via ultraviolet (UV) photoinitiated copolymerization of ionic liquid (IL) monomers on a fused silica support is described. The copolymerization of monocationic and dicationic IL cross-linkers was performed "on-fiber" using UV initiated free radical polymerization. Mixtures composed of various amounts of the IL cross-linker, UV initiator (DAROCUR 1173), and IL monomer were dip-coated onto an etched and derivatized fused silica support and placed in a high-capacity UV reactor. The method requires no organic dispersive solvent and is much more rapid compared to traditional methods in which polymeric ionic liquid (PIL) sorbent coatings are prepared by 2,2'-azobis(2-methylpropionitrile) (AIBN)-initiated polymerization. Two ionic liquid-based cross-linkers, namely, 1,8-di (3-vinylimidazolium) octane dibromide ([(VIM)(2)C(8)] 2[Br]) and 1,12-di (3-vinylimidazolium) dodecane dibromide ([(VIM)(2)C(12)] 2[Br]), were copolymerized with an ionic liquid monomer, 1-vinyl-3-hexylimidazolium chloride ([VHIM][Cl]), to produce polar cross-linked PIL-based SPME sorbent coatings. The cross-linking and immobilization of these coatings make them particularly applicable in direct immersion SPME. The coatings were applied in the extraction of polar analytes, including alcohols, aldehydes, and esters, from aqueous solutions using headspace and direct immersion SPME gas chromatography mass spectrometry (GC/MS). Compared to linear PIL-based sorbent coatings containing the same anions, the cross-linked PIL-based coatings exhibited higher thermal stability and lower bleed, making them ideal for GC/MS. Recovery experiments were performed in deionized, well, and river water. The structural integrity of the sorbent coatings, as well as their analytical precision, was not diminished after 90 extractions from complex samples using headspace and direct immersion SPME.

We assessed the ability of muscle-derived stem cells (MDSC) to differentiate into smooth muscle cells (SMC) and their potential to promote the regeneration of smooth muscle with a vessel extracellular matrix (VECM) for tissue engineering of the ureter. MDSC were isolated, proliferated, and identified by flow cytometry. SMC phenotype differentiation was induced with a smooth muscle induction medium. Gene expression was evaluated by real-time quantitative polymerase chain reaction (PCR) and Western blot studies. The VECM was obtained by a decellularization process, and cytotoxic effects were evaluated by exposing the induced cells to a VECM extract. The induced cells were seeded onto VECM in vitro for 1 week, and then the compound grafts were used for ureter reconstitution in vivo. The grafts were obtained for histological studies at 2, 4, 8, and 16 weeks post-operation. Intravenous urography was used to evaluate renal function and ureteral patency. Flow cytometry demonstrated that the MDSC expressed Sca-1 and desmin, but did not express CD45. After induction, SMC phenotype gene expression was confirmed in the induced cells by real-time quantitative PCR and Western blot studies. VECM exhibited a nontoxic effect on the induced cells in vitro. At 16 weeks post-operation, a histological evaluation showed that multilayered urothelium and organized muscle fiber bundles had formed in the grafts. Intravenous urography demonstrated no evidence of ureteral stricture or hydroureteronephrosis. These results demonstrate that MDSC can be induced into SMC and that this was useful for promoting regeneration of smooth muscles for ureter tissue engineering.

Three structurally different ionic liquids (ILs), namely 1-butyl-3-methylimidazolium chloride ([BMIM][Cl]), 1-(6-hydroxyethyl)-3-methylimidazolium chloride ([HeOHMIM][Cl]) and 1-benzyl-3-(2-hydroxyethyl)imidazolium bromide ([BeEOHIM][Br]), were applied as extraction solvents using in situ dispersive liquid-liquid microextraction (in situ DLLME) for the preconcentration of two microcystin variants, microcystin-RR (MC-RR) and microcystin-LR (MC-LR) from aqueous samples. Extraction parameters including sample solution pH, ratio of IL to metathesis reagent, sample volume, IL quantity, and salt concentration were optimized to achieve the best extraction efficiency. The [BeEOHIM][Br] IL, which contains both an aromatic moiety and a hydroxyl group within its chemical structure, exhibited superior extraction efficiency compared to the other two ILs. The analytical performance of the [BeEOHIM][Br] IL as an extraction solvent for in situ DLLME of microcystins was investigated using HPLC-UV and HPLC-MS. The limits of detection (LODs) for MC-RR and MC-LR were 0.7μgL(-1) using UV detection with a linear range from 1 to 50μgL(-1). The separation method was successfully adapted for ESI-MS/SIM detection, wherein the LODs for MC-RR and MC-LR were greatly improved to 0.005 and 0.003μgL(-1), respectively. The accuracy of the method was demonstrated by examining the relative recovery using tap water and river water and produced recoveries ranging from 45.0 to 109.7% and from 46.3 to 103.2%, respectively.

Ionic liquids (ILs) are a class of organic salts with unique properties and have been shown to be particularly useful for the development and improvement of sample preparation and detection methodologies. ILs and polymeric ionic liquids (PILs) have played an important role in the analysis of environmental samples. These compounds have been widely applied in various microextraction techniques due to their tunable analyte selectivity, low volatility, good thermal stability, as well as variable viscosity and solvent miscibility. The unique properties also make ILs and PILs an emerging class of stationary phases in gas chromatography (GC). IL/PIL-based stationary phases exhibit excellent applicability in multidimensional GC separations. This chapter will highlight the applications of ILs and PILs in a number of microextraction techniques in addition to their utilization in separation science with a particular emphasis on their use in environmental analysis.

Numerous studies have shown that human endogenous retrovirus W family (HERV-W) envelope gene (env) is related to various diseases but the underlying mechanism has remained poorly understood. Our previous study showed that there was abnormal expression of HERV-W env in sera of patients with schizophrenia. In this paper, we reported that overexpression of the HERV-W env elevated the levels of small conductance Ca(2+)-activated K(+) channel protein 3 (SK3) in human neuroblastoma cells. Using a luciferase reporter system and RNA interference method, we found that functional cAMP response element site was required for the expression of SK3 triggered by HERV-W env. In addition, it was also found that the SK3 channel was activated by HERV-W env. Further study indicated that cAMP response element-binding protein (CREB) was required for the activation of the SK3 channel. Thus, a novel signaling mechanism of how HERV-W env influences neuronal activity and contributes to mental illnesses such as schizophrenia was proposed.

Human endogenous retrovirus (HERV) accounts for ∼8% of the human genome. Recent studies have reported that multiple HERV genes and long terminal repeats (LTRs) are involved in human tumorigenesis. Here we demonstrated that HERV-W env (syncytin-1) was overexpressed in 75.6% (62/82) of urothelial cell carcinoma (UCC) tissues of the bladder compared with only 6.1% (5/82) of matched tumor-adjacent tissues (P0.001). Syncytin-1 overexpression increased proliferation and viability of immortalized human uroepithelial cells. Colony-formation experiments and in-vivo tumor xenografts suggested that syncytin-1 overexpression had oncogenic potential. Syncytin-1 3'-LTR mutations (142TC and 277AG) were present in 87.8% (72/82) of UCC tissues. Normal 3'-LTR was found in 12.2% (10/82) of UCC tissues compared with 95.1% (78/82) of matched tumor-adjacent tissues (P0.001). Interestingly, 3'-LTR mutations were significantly associated with syncytin-1 overexpression. Luciferase assay and expression analysis revealed that 3'-LTR mutations, especially the 142TC mutation, enhanced the syncytin-1 promoter activity and expression. In-silico analysis, electrophoretic mobility shift assays and chromatin immunoprecipitation assays demonstrated the binding of c-Myb to 3'-LTRs when the mutations occurred. This alternative interaction was found to be dependent on 142TC mutation. C-Myb activated syncytin-1 promoter activity and expression by binding to mutant 3'-LTRs. Taken together, these data indicate that syncytin-1 overexpression may be an indicator of UCC risk. The 3'-LTR mutations may upregulate syncytin-1 expression, enabling it to participate in UCC tumorigenesis and development by interacting with c-Myb.

Objective: Retrovirus has been suggested as one of agents involved in the development of schizophrenia. In the present study, we examined the role of the human endogenous retrovirus W family (HERV-W) env gene in the etiopathogenesis of recent-onset schizophrenia, using molecular and epidemiological approaches. Methods: Nested RT-PCR was used to detect the messenger RNA (mRNA) of the HERV-w env gene in plasmas. Quantitative real-time polymerase chain reaction (PCR) was employed to detect the viral reverse transcriptase activity in human sera. Human U251 glioma cells were used to study the potential role of the HERV-W env gene in the etiopathogenesis of recent-onset schizophrenia. Results: We identified genes with mRNA sequences homologous to HERV-W env gene from plasmas of 42 out of 118 individuals with recent-onset schizophrenia but not from any of 106 normal persons (P < .01, t test). Quantitative real-time PCR showed a significantly increase in the reverse transcriptase activity in the sera of patients (by 35.59%) compared with controls (by 2.83%) (P < .05, t test). Overexpression of HERV-w env in human U251 glioma cells upregulated brain-derived neurotrophic factor (BDNF), an important schizophrenia-associated gene, neurotrophic tyrosine kinase receptor type 2 (NTRK2, also called TrkB), and dopamine receptor D3 and increased the phosphorylation of cyclic adenosine monophosphate response element–binding (CREB) protein. BDNF promoter reporter gene assays showed that the HERV-W env triggers BDNF production in human U251 glioma cells. Using gene knockdown, we found that CREB is required for the expression of BDNF that is regulated by env. Conclusion: Our data revealed that the transcriptional activation of HERV is associated with the development of schizophrenia in some patients and indicated that HERV-W env regulates the expression of schizophrenia-associated genes. This report is the first to elucidate the signaling pathway responsible for the upregulation of HERV-W env–triggered BDNF. Our study provides new evidence for the involvement of HERV-W in the central nervous system, which will benefit the diagnosis and treatment of the devastating schizophrenia and related disorders.