Enzymatic functionalization of decellularized tilapia skin scaffolds with enhanced skin regeneration.

The decellularized tilapia skin (dTS) has gained significant attention as a promising material for tissue regeneration due to its ability to provide unique structural and functional components that support cell growth, adhesion, and proliferation. However, the clinical application of dTS is limited by its low mechanical strength and rapid biodegradability. Herein, we prepare a novel RGD (arginine-glycine-aspartic acid) functionalized dTS scaffold (dTS/RGD) by using transglutaminase (TGase) crosslinking. The developed dTS/RGD scaffold possesses excellent properties, including a medium porosity of ∼59.2%, a suitable degradation rate of approximately 80% over a period of two weeks, and appropriate mechanical strength with a maximum tensile stress of ∼46.36 MPa which is much higher than that of dTS (∼32.23 MPa). These properties make the dTS/RGD scaffold ideal for promoting cell adhesion and proliferation, thereby accelerating skin wound healing in a full-thickness skin defect model. Such an enzymatic cross-linking strategy provides a favorable microenvironment for wound healing and holds great potential for application in skin regeneration engineering.

SERS determination of sodium saccharin content on the tipping paper of cigarettes using AgNP substrates prepared with a USB-power supply device.

A novel magnetic agitating heater powered by a USB port has been developed and applied to synthesize silver colloid substrate for surface-enhanced Raman scattering (SERS) detection of sodium saccharin content on the tipping paper of cigarettes. The heater device allows the convenient synthesis of the Ag colloid, and the reaction can be completed on-site in 15 min under mild conditions. The on-site synthesis of SERS substrate effectively avoided the need for storage and concerns regarding the poor stability and short lifespan of colloid substrates. The results demonstrated that the substrate obtained with the device could achieve SERS detection of Rhodamine 6G (R6G) at as low as 10-9 mol L-1 while maintaining a stable intensity with a relative standard deviation (RSD) of 5.52% (n = 5). Using the prepared substrate at the optimal conditions, the limit of detection of sodium saccharin (SS) was 1 mg L-1. By introducing an internal standard KSCN, a linear relationship was observed between the relative intensity at 708 cm-1 and the concentration of the SS in a range of 20-100 mg L-1 (R2 = 0.98). With the developed method, SS was directly extracted from the cigarette paper by immersing it in water, and the extracted solution was subsequently detected. The quantitative spike-recoveries were in the range of 95.5-116.7%, with RSD between 2.3-12.6%. The whole detection procedure including the on-site substrate preparation, took only about 30 min. This work opens new avenues for colloidal synthesis, and the detection method of SS on the cigarette paper also holds great promise in food safety applications.

Disassembling ability of lipopeptide promotes the antibacterial activity.

Lipopeptides have become one of the most potent antibacterial agents, however, there is so far no consensus about the link between their physic-chemical properties and biological activity, in particular their inherent aggregation propensity and antibacterial potency. To this end, we here de novo design a series of lipopeptides (CnH(2n-1)O-(VVKK)2V-NH2), in which an alkyl chain is covalently attached onto the N-terminus of a short cationic peptide sequence with an alternating pattern of hydrophobic VV (Val) and positively charged KK (Lys) motifs. By varying the alkyl chain length (ortho-octanoic acid (C8), lauric acid (C12), and palmitic acid (C16)), the lipopeptides show distinct physicochemical properties and self-assembly behaviors, which have great effect on their antibacterial activities. C8H15O-(VVKK)2V-NH2, which contains the lowest hydrophobicity and surface activity has the lowest antibacterial activity. C12H23O-(VVKK)2V-NH2 and C16H31O-(VVKK)2V-NH2 both have high hydrophobicity and surface activity, and self-assembled into long nanofibers. However, the nanofibers formed by C12H23O-(VVKK)2V-NH2 disassembled by dilution, resulting in its high antibacterial activity via bacterial membrane disruption. Comparatively, the nanofibers formed by C16H31O-(VVKK)2V-NH2 were very stable, which can closely attach on bacterial surface but not permeate bacterial membrane, leading to its low antibacterial activity. Thus, the stability other than the morphologies of lipopeptides' nanostructures contribute to their antibacterial ability. Importantly, this study enhances our understanding of the antibacterial mechanisms of self-assembling lipopeptides that will be helpful in exploring their biomedical applications.

Amyloid-like aggregates of short self-assembly peptide selectively induce melanoma cell apoptosis.

With the rising global incidence of melanoma, new anti-melanoma drugs with low-inducing drug resistance and high selectivity are in urgent need. Inspired by the physiological events in which fibrillar aggregates formed by amyloid proteins are toxic to normal tissues, we here rationally design a tyrosinase responsive peptide, I4K2Y* (Ac-IIIIKKDopa-NH2). Such peptide self-assembled into long nanofibers outside the cells, while it was catalyzed into amyloid-like aggregates by tyrosinase which was rich in melanoma cells. The newly formed aggregates concentrated around the nucleus of melanoma cells, blocking the exchange of biomolecules between the nucleus and cytoplasm and finally leading to cell apoptosis via the S phase arrest in cell cycle distribution and dysfunction of mitochondria. Furthermore, I4K2Y* effectively inhibited B16 melanoma growth in a mouse model but with minimal side effects. We believe that the strategy of combining the usage of toxic amyloid-like aggregates and in-situ enzymatic reactions by specific enzymes in tumor cells will bring profound implications for designing new anti-tumor drugs with high selectivity.

Genomic and Immunological Characterization of Pyroptosis in Lung Adenocarcinoma.

Pyroptosis is a programmed cell death that may either promote or hinder cancer growth under different circumstances. Pyroptosis-related genes (PRGs) could be a useful target for cancer therapy, and are uncommon in lung adenocarcinoma (LUAD). The expression profiles, mutation data and clinical information of LUAD patients were included in this study. A pyroptosis-related prognostic risk score (PPRS) model was constructed by performing Cox regression, weighted gene co-expression network analysis (WGCNA), and least absolute shrinkage and selection operator (LASSO) analysis to score LUAD patients. Somatic mutation and copy number variation (CNV), tumor immunity, and sensitivity to immunotherapy/chemotherapy were compared between different PPRS groups. Clinical parameters of LUAD were combined with PPRS to construct a decision tree and nomogram. Red module was highly positively correlated with pyroptosis. Seven genes (FCRLB, COTL1, GNG10, CASP4, DOK1, CCR2, and AQP8) were screened from the red module to construct a PPRS model. Significantly lower overall survival (OS), higher incidence of somatic mutation and CNV, elevated infiltration level of the immune cell together with increased probability of immune escape were observed in LUAD patients with higher PPRS, and were more sensitive to Cisplatin, Docetaxel, and Vinorelbine. We constructed a new PPRS model for patients with LUAD. The model might have clinical significance in the prediction of the prognosis of patients with LUAD and in the efficacy of chemotherapy and immunotherapy.

Metformin promotes anticancer activity of NK cells in a p38 MAPK dependent manner.

Metformin, a drug prescribed to treat type 2 diabetes, has been reported to possess antitumor activity via immunity activation. However, the influence of metformin on natural killer (NK) cells is not fully understood. Here, we investigated whether metformin exerts a potent anticancer effect by activating NK cells. The results showed that sustained exposure to metformin enhances the cytolytic activity of NK-92 cells. Moreover, this enhancement of cytotoxicity by metformin was also observed in NK cells from healthy peripheral blood and cancer patient ascites. Mechanistically, metformin induced activation of the JAK1/2/3/STAT5 and AKT/mTOR pathways in a p38 MAPK-dependent manner rather than an AMPK-dependent manner. In vivo experiments, metformin also improved cancer surveillance of NK cells in mouse models of lymphoma clearance and metastatic melanoma. Additionally, combination treatment with metformin and anti-PD-1 antibodies increased the therapy response rates of B16F10 melanoma. Moreover, metformin treatment increased NK cell and T cell infiltration in tumors. Therefore, these results provide a deeper understanding of metformin on the effector function of NK cells and will contribute to the development and applications of metformin in cancer treatment strategies.

MRTF-A-NF-κB/p65 axis-mediated PDL1 transcription and expression contributes to immune evasion of non-small-cell lung cancer via TGF-ß.

PD-L1 is abnormally regulated in many cancers and is critical for immune escape. Fully understanding the regulation of PD-L1 expression is vital for improving the clinical efficacy of relevant anticancer agents. TGF-ß plays an important role in the low reactivity of PD-1/PD-L1 antibody immunotherapy. However, it is not very clear whether and how TGF-ß affects PD-L1 expression. In the present study, we show that TGF-ß upregulates the expression of the transcriptional coactivator MRTF-A in non-small-cell lung cancer cells, which subsequently interacts with NF-κB/p65 rather than SRF to facilitate the binding of NF-κB/p65 to the PDL1 promoter, thereby activating the transcription and expression of PD-L1. This leads to the immune escape of NSCLC cells. This process is dependent on the activation of the TGF-ß signaling pathway. In vivo, inhibition of MRTF-A effectively suppresses the growth of lung tumor syngrafts with enrichment of NK and T cells in tumor tissue. Our study defines a new signaling pathway that regulates the transcription and expression of PD-L1 upon TGF-ß treatment, which may have a significant impact on research into the application of immunotherapy in treating lung cancer.

PGK1-coupled HSP90 stabilizes GSK3ß expression to regulate the stemness of breast cancer stem cells.

OBJECTIVE: Glycogen synthase kinase-3ß (GSK3ß) has been recognized as a suppressor of Wnt/ß-catenin signaling, which is critical for the stemness maintenance of breast cancer stem cells. However, the regulatory mechanisms of GSK3ß protein expression remain elusive. METHODS: Co-immunoprecipitation and mass spectral assays were performed to identify molecules binding to GSK3ß, and to characterize the interactions of GSK3ß, heat shock protein 90 (Hsp90), and co-chaperones. The role of PGK1 in Hsp90 chaperoning GSK3ß was evaluated by constructing 293T cells stably expressing different domains/mutants of Hsp90α, and by performing a series of binding assays with bacterially purified proteins and clinical specimens. The influences of Hsp90 inhibitors on breast cancer stem cell stemness were investigated by Western blot and mammosphere formation assays. RESULTS: We showed that GSK3ß was a client protein of Hsp90. Hsp90, which did not directly bind to GSK3ß, interacted with phosphoglycerate kinase 1 via its C-terminal domain, thereby facilitating the binding of GSK3ß to Hsp90. GSK3ß-bound PGK1 interacted with Hsp90 in the "closed" conformation and stabilized GSK3ß expression in an Hsp90 activity-dependent manner. The Hsp90 inhibitor, 17-AAG, rather than HDN-1, disrupted the interaction between Hsp90 and PGK1, and reduced GSK3ß expression, resulting in significantly reduced inhibition of ß-catenin expression, to maintain the stemness of breast cancer stem cells. CONCLUSIONS: Our findings identified a novel regulatory mechanism of GSK3ß expression involving metabolic enzyme PGK1-coupled Hsp90, and highlighted the potential for more effective cancer treatment by selecting Hsp90 inhibitors that do not affect PGK1-regulated GSK3ß expression.

Synergistic combination therapy of lung cancer using lipid-layered cisplatin and oridonin co-encapsulated nanoparticles.

Lung cancer treatment using cisplatin (DDP) in combination with other drugs are effective for the treatment of non-small cell lung cancer (NSCLC). The aim of this study was to prepare a layer-by-layer nanoparticles (NPs) for the co-loading of DDP and oridonin (ORI) and to evaluate the antitumor activity of the system in vitro and in vivo. Novel DDP and ORI co-loaded layer-by-layer NPs (D/O-NPs) were constructed. The mean diameter, surface change stability and drug release behavior of NPs were evaluated. In vitro cytotoxicity of D/O-NPs was investigated against DDP resistant human lung cancer cell line (A549/DDP cells), and in vivo anti-tumor efficiency of D/O-NPs was tested on mice bearing A549/DDP cells xenografts. D/O-NPs have a diameter of 139.6 ± 4.4 nm, a zeta potential value of +13.8 ± 1.6 mV. D/O-NPs could significantly enhance in vitro cell toxicity and in vivo antitumor effect against A549/DDP cells and lung cancer animal model compared to the single drug loaded NPs and free drugs. The results demonstrated that the D/O-NPs could be used as a promising lung cancer treatment system.

Comprehensive analyses of PBRM1 in multiple cancer types and its association with clinical response to immunotherapy and immune infiltrates.

BACKGROUND: The prognostic value of polybromo 1 (PBRM1) gene mutations in clear cell renal carcinoma (CCRCC) with anti-programmed death-ligand 1 (PD-L1) therapy remains controversial, and few studies have reported the impact of PBRM1 mutations in other cancer types. METHODS: The patient information was obtained from cBioPortal and the Tumor Immune Estimation Resource (TIMER) databases. Mann-Whitney U test were used for correlation analysis. For survival analyses, Kaplan-Meier survival curves were used and compared using the log-rank test. Cox's regression model was used to perform univariable and multivariable analyses. RESULTS: Our study, for the first time, performed comprehensive analyses of PBRM1 mutation frequency, PBRM1 expression, relationship of PBRM1 mutations with clinical benefit from immunotherapy, and PBRM1 expression with immune infiltrates in diverse cancer types. The results showed that the expression of PBRM1 was significantly lower in diverse cancer types compared with normal tissues. Based on multivariable analysis, PBRM1 mutations trended towards worse clinical outcomes from anti-PD-L1 in CCRCC, lung adenocarcinoma (LUAD), bladder urothelial carcinoma (BLCA), and skin cutaneous melanoma (SKCM), and a significant association was observed in LUAD and BLCA. PBRM1 mutations were associated with higher TMB in diverse cancer types and significant associations were observed in LUAD and BLCA. The expression of PBRM1 was found to positively correlate with immune infiltrates in diverse cancer types. CONCLUSIONS: Our findings suggested caution in starting immunotherapy alone in PBRM1 mutant patients. Further studies are needed to improve treatment for PBRM1 mutant patients.

Rapid Hemostasis Resulting from the Synergism of Self-Assembling Short Peptide and O-Carboxymethyl Chitosan.

The development of novel hemostatic agents with distinct modes of action from traditional ones remains a formidable challenge. Self-assembling peptide hydrogels have emerged as a new hemostatic material, not only because of their inherent biocompatibility and biodegradability but also their designability. Especially, rational molecular design can make peptides and their hydrogelation responsive to biological cues. In this study, we demonstrated that transglutaminase-catalyzed reactions not only occurred among designed short peptide I3QGK molecules but also between the peptide and a natural polysaccharide O-carboxymethyl chitosan. Because Factor XIII in the blood can rapidly convert into activated transglutaminase (Factor XIIIa) upon bleeding, these enzymatic reactions, together with the electrostatic attraction between the two hemostatic agents, induced a strong synergetic effect in promoting hydrogelation, blood coagulation, and platelet adhesion, eventually leading to rapid hemostasis. The study presents a promising strategy for developing alternative hemostatic materials and methods.

LncRNA SLC7A11-AS1 Contributes to Lung Cancer Progression Through Facilitating TRAIP Expression by Inhibiting miR-4775.

PURPOSE: Long non-coding RNAs (lncRNAs) are important regulators of lung cancer. This article introduced a novel lncRNA, SLC7A11-AS1, whose effects on lung cancer development have been explored. METHODS: Lung cancer tissues and normal tissues of 47 patients were collected. Bronchial epithelial cell line (BEAS-2B) and lung cancer cell lines (H520, H596, A549 and H1299) were cultured. H1299 and A549 cells were transfected with siSLC7A11-AS1 or siNC. The proliferation, migration and invasion of H1299 and A549 cells were detected by CCK-8 assay and Transwell experiment. Caspase-3 activity in H1299 and A549 cells was researched using caspase-3 activity detection kit. Dual-luciferase reporter gene assay and RNA pull-down assay were performed to explore the relationship between SLC7A11-AS1 and miR-4775. SLC7A11-AS1, miR-4775 and TRAIP mRNA expressions in tissues/cells were detected by qRT-PCR. RESULTS: The up-regulated SLC7A11-AS1 in lung cancer patients was associated with metastasis and advanced tumor stage (P < 0.05). SLC7A11-AS1 was significantly up-regulated in lung cancer cells (P < 0.05). Silencing of SLC7A11-AS1 prominently inhibited H1299 and A549 cells proliferation, migration and invasion in vitro (P < 0.05). SLC7A11-AS1 acted as a sponge to inhibit miR-4775 expression in H1299 and A549 cells. Meanwhile, TRAIP expression in H1299 and A549 cells was directly and negatively regulated by miR-4775. Inhibition of miR-4775 or overexpression of TRAIP in H1299 and A549 cells remarkably reversed the reduced proliferation, migration and invasion induced by SLC7A11-AS1 silencing (P < 0.05). CONCLUSION: SLC7A11-AS1 promoted lung cancer development by enhancing TRAIP expression via suppressing miR-4775.

Alpinumisoflavone radiosensitizes esophageal squamous cell carcinoma through inducing apoptosis and cell cycle arrest.

Radiotherapy remains a mainstream treatment for patients with unresectable and locally advanced esophageal squamous cell carcinoma (ESCC). However, intrinsic radioresistance of ESCC tumors has largely compromised the efficacy of radiotherapy. The following study investigates the potential radiosensitizing effect of alpinumisoflavone (AIF) and explores its underlying mechanisms in ESCC. Briefly, our results showed that AIF could significantly increase radiosensitivity of ESCC cells both in vitro and in vivo, by increasing the effect of AIF on irradiation-induced DNA damage, apoptosis and cell cycle arrest. Mechanically, AIF aggravated irradiation-induced ROS generation in ESCC cells, which occurred via suppressing the expression of nuclear transcription factor Nrf2 and Nrf2-driven antioxidant molecule NQO-1 and HO-1. Collectively, we concluded that AIF functions as a potent radiosensitizer in human ESCC.

miR-107 regulates cisplatin chemosensitivity of A549 non small cell lung cancer cell line by targeting cyclin dependent kinase 8.

Previous studies demonstrated that the acquired drug resistance of non-small cell lung cancer (NSCLC) was related to deregulation of miRNAs. However, the effects of miR-107 and the mechanism through which miR-107 affects the cisplatin chemoresistance in NSCLC have not been reported. TaqMan RT-PCR or Western blot assay was performed to detect the expression of mature miR-107 and cyclin dependent kinase 8 (CDK8) protein. The viabilities of treated cells were analyzed using MTT assay. We found that the expression level of miR-107 in A549 cells was significantly lower than that in normal human bronchial epithelial cells (0.45 ± 0.26 vs. 1.00 ± 0.29, P = 0.032). The MTT assay showed that the A549 cells transfected with miR-107 mimics were significantly more sensitive to the therapy of cisplatin than control cells. A549 cells transfected with miR-107 mimics showed a decreased CDK8 protein expression. Downregulation of CDK8 expression by siRNAs, A549 cells became more sensitive to the therapy of cisplatin. In addition, the enhanced growth-inhibitory effect by the miR-107 mimic transfection was enhanced after the addition of CDK8 siRNA. In conclusion, the present study provides the first evidence that miR-107 plays a key role in cisplatin resistance by targeting the CDK8 protein in NSCLC cell lines, suggesting that miR-107 can be used to predict a patient's response to chemotherapy as well as serve as a novel potential maker for NSCLC therapy.

Video-assisted thoracoscopic surgery for treatment of early- stage non-small cell lung cancer.

OBJECTIVES: To evaluate the safety, efficacy, and invasiveness of lobectomy by video-assisted thoracoscopic surgery (VATS) in the treatment of stage I/II non-small cell lung cancer (NSCLC). METHODS: A total of 148 patients presenting with Stage I or II NSCLC were enrolled into our study, comprising 71 who underwent VATS and 77 patients undergoing conventional thoracotomic lobectomy, in combination with systematic lymph node resection. RESULTS: It was found that VATS was superior to conventional thoracotomy in terms of the duration of surgery, intraoperative blood loss, frequency of the need to administer postoperative analgesia, thoracic intubation indwelling time, post-operative hospital stay, and survival rate (P<0.05). We saw no obvious difference in the number of resected lymph nodes with either approach. CONCLUSIONS: VATS lobectomy is a safe and reliable surgical approach for the treatment of Stage I/II NSCLC, characterized by significantly minimal invasiveness, rapid post-operative recovery, and markedly lower loss of blood.