Targeted inhibition of the HNF1A/SHH axis by triptolide overcomes paclitaxel resistance in non-small cell lung cancer.

Paclitaxel resistance is associated with a poor prognosis in non-small cell lung cancer (NSCLC) patients, and currently, there is no promising drug for paclitaxel resistance. In this study, we investigated the molecular mechanisms underlying the chemoresistance in human NSCLC-derived cell lines. We constructed paclitaxel-resistant NSCLC cell lines (A549/PR and H460/PR) by long-term exposure to paclitaxel. We found that triptolide, a diterpenoid epoxide isolated from the Chinese medicinal herb Tripterygium wilfordii Hook F, effectively enhanced the sensitivity of paclitaxel-resistant cells to paclitaxel by reducing ABCB1 expression in vivo and in vitro. Through high-throughput sequencing, we identified the SHH-initiated Hedgehog signaling pathway playing an important role in this process. We demonstrated that triptolide directly bound to HNF1A, one of the transcription factors of SHH, and inhibited HNF1A/SHH expression, ensuing in attenuation of Hedgehog signaling. In NSCLC tumor tissue microarrays and cancer network databases, we found a positive correlation between HNF1A and SHH expression. Our results illuminate a novel molecular mechanism through which triptolide targets and inhibits HNF1A, thereby impeding the activation of the Hedgehog signaling pathway and reducing the expression of ABCB1. This study suggests the potential clinical application of triptolide and provides promising prospects in targeting the HNF1A/SHH pathway as a therapeutic strategy for NSCLC patients with paclitaxel resistance. Schematic diagram showing that triptolide overcomes paclitaxel resistance by mediating inhibition of the HNF1A/SHH/ABCB1 axis.

Characteristics of CD4-1 gene and its immune responses against Aeromonas veronii infection by activating NF-κB signaling in Qihe crucian carp Carassius auratus.

CD4-1 found in bony fish contains four extracellular immunoglobulin (Ig)-like domains similar to that of mammalian CD4, which is crucial for the activation of CD4+ helper T-cell. However, there is limited knowledge regarding the molecular markers, immune functions and regulation mechanism of CD4-1 in teleosts due to their vast diversity. In this study, we cloned and characterized two isoforms of Qihe crucian carp CD4-1, designated as CaCD4-1.1 and CaCD4-1.2. We further explored their expression responses upon stimulation with Aeromonas veronii, and the regulation of their immune responses against A. veronii by NF-κB. The ORF of CaCD4-1.1 and CaCD4-1.2 cDNA encoded 477 and 466 amino acids, respectively. Both proteins contained seven conserved cysteine residues in the extracellular domain, and a CCC motif in their cytoplasm, respectively. However, CaCD4-1.1 exhibited a relatively limited similarity with CaCD4-1.2 in the ectodomain. The quantitative real-time polymerase chain reaction (qRT-PCR) analysis revealed that the mRNA expression of CaCD4-1.1 and CaCD4-1.2 exhibited differential constitutive expression across all examined tissues. Furthermore, the expression level of CD4-1.2 was higher than that of CD4-1.1 in the gills, head kidney, and spleen of Qihe crucian carp subjected to A. veronii challenge, while it was lower in the trunk kidney. Inhibition of NF-κB activity resulted in a decrease in the expression levels of CD4-1.1 and CD4-1.2 mRNA in the gill, while inducing an increase in expression levels in the spleen, in accordance with the observed ultrastructural changes in both organs. Interestingly, the impact of NF-κB on the mRNA expression level of CD4-1.1 appears to be stronger than that of CD4-1.2. Our results suggest that CaCD4-1.1 and CaCD4-1.2 could be expressed on T cells and antigen-sampling cells that exhibit similar characteristics to mammalian M cells, respectively, and differentially regulated by NF-κB in adaptive immune responses against bacterial infection. This research contributes to a better understanding of the crucial role of CD4-1 in the immune response of Qihe crucian carp and provide novel insights for the prevention and treatment of fish diseases in aquaculture.

The role of CcPTGS2a in immune response against Aeromonas hydrophila infection in common carp (Cyprinus carpio).

Prostaglandin-endoperoxide synthase 2 (PTGS2), a crucial enzyme in prostaglandin synthesis, catalyzes the conversion of arachidonic acid to prostaglandins and plays a significant role in the inflammatory response. This investigation aimed to determine the regulatory role of PTGS2a in the innate immune response to bacterial infection in fish. To achieve this objective, the CcPTGS2a gene was identified and characterized in common carp (Cyprinus carpio), and its function in immune defense was investigated. According to the sequence and structural analysis results, CcPTGS2a had an open reading frame of 1806 bp that encoded 602 amino acids. It was estimated that the protein's theoretical molecular weight was 69.0 kDa, and its isoelectric point was 8.10. The structure of CcPTGS2a was observed to be conserved, with an epidermal growth factor domain and a peroxidase domain present. Moreover, the amino acid sequence of CcPTGS2a exhibited significant homology with the amino acid sequences of several fish species. CcPTGS2a mRNA was detected in the healthy tissues of common carp, with higher expression in the head kidney, spleen, gills, and liver. Following the challenges with Aeromonas hydrophila and lipopolysaccharide, CcPTGS2a mRNA showed unique geographic and temporal expression patterns, with significant increases detected in the head kidney, gills, spleen, and liver. Additionally, the recombinant CcPTGS2a protein exhibited detectable bacterial binding to various bacteria. As determined by subcellular localization analysis, CcPTGS2a was predominantly localized in the nucleus and cytoplasm. Furthermore, it was discovered that the overexpression of CcPTGS2a stimulated the up-regulation of ferroptosis-related genes and inflammatory cytokine mRNA expression in fish and EPC (Epithelioma papulosum cyprinid) cells while concurrently reducing the bacterial load of A. hydrophila. In contrast, the interference of CcPTGS2a decreased the mRNA expression of ferroptosis-related genes and inflammatory cytokines in fish and EPC cells and increased the bacterial load of A. hydrophila. Notably, A. hydrophila stimulation resulted in the up-regulation of CcPTGS2a protein expression in EPC cells. These results suggested that CcPTGS2a was involved in the immune response to bacterial infections in common carp.

Two CcCCL19bs orchestrate an antibacterial immune response in Yellow River carp (Cyprinus carpio haematopterus).

Chemokines are a group of chemotactic cytokines with an essential role in homeostasis as well as immunity via specific G protein-coupled receptors and atypical receptors. In our study, two Yellow River carp (Cyprinus carpio haematopterus) CCL19b genes (CcCCL19bs), tentatively named CcCCL19b_a and CcCCL19b_b, were cloned. The open reading frames (ORFs) of CcCCL19b_a and CcCCL19b_b were both 333 bp that encoded a 12 kDa protein with 110 amino acid residues. CcCCL19bs contained a signal peptide and a SCY domain with four typical conserved cysteine residues. The two CcCCL19b proteins shared high similarities with each other in both secondary and three-dimensional structure. Phylogenetic analysis showed that CcCCL19bs and other CCL19bs from tetraploid cyprinid fish were clustered into one clade. CcCCL19bs were highly expressed in gill and intestine in healthy fish, and a significant up-regulation of gene expression after Aeromonas hydrophila infection and poly(I:C) stimulation was observed in gill, liver, and head kidney. Furthermore, chemotaxis and antibacterial activity of CcCCL19bs were studied. The results indicated that recombinant CcCCL19b_a and CcCCL19b_b protein (rCcCCL19b_a and rCcCCL19b_b) exhibited significant attraction to primary head kidney leukocytes (HKLs). Meanwhile, both of rCcCCL19bs could promote the proliferation of HKLs, and significantly up-regulate the expressions of IL-1ß, CCR7, and IL-6, and down-regulate the expression of IL-10 in primary HKLs. In vitro, rCcCCL19bs could bind and aggregate A. hydrophila and Staphylococcus aureus. The rCcCCL19bs exhibited significant antibacterial activity against A. hydrophila, but not S. aureus. Moreover, they inhibited the growth of A. hydrophila and S. aureus. In vivo, overexpression of CcCCL19bs contributed to the bacterial clearance. These studies suggested that CcCCL19bs orchestrate an antibacterial immune response.

Activation Mechanism of CcGSDMEb-1/2 and Regulation for Bacterial Clearance in Common Carp (Cyprinus carpio).

Gasdermin E (GSDME), to date, is considered the only direct executor of the pyroptosis process in teleost and plays an important role in innate immunity. In common carp (Cyprinus carpio), there contains two pairs of GSDME (GSDMEa/a-like and GSDMEb-1/2), and the pyroptotic function and regulation mechanism of GSDME still remain unclear. In this study, we identified two GSDMEb genes of common carp (CcGSDMEb-1/2), which contain a conserved N-terminal pore-forming domain, C-terminal autoinhibitory domain, and a flexible and pliable hinge region. We investigated the function and mechanism of CcGSDMEb-1/2 in association with inflammatory and apoptotic caspases in Epithelioma papulosum cyprinid cells and discovered that only CcCaspase-1b could cleave CcGSDMEb-1/2 through recognizing the sites 244FEVD247 and 244FEAD247 in the linker region, respectively. CcGSDMEb-1/2 exerted toxicity to human embryonic kidney 293T cells and bactericidal activity through its N-terminal domain. Interestingly, after i.p. infection by Aeromonas hydrophila, we found that CcGSDMEb-1/2 were upregulated in immune organs (head kidney and spleen) at the early stage of infection, but downregulated in mucosal immune tissues (gill and skin). After CcGSDMEb-1/2 were knocked down and overexpressed in vivo and in vitro, respectively, we found that CcGSDMEb-1/2 could govern the secretion of CcIL-1ß and regulate the bacterial clearance after A. hydrophila challenge. Taken together, in this study, it was demonstrated that the cleavage mode of CcGSDMEb-1/2 in common carp was obviously different from that in other species and played an important role in CcIL-1ß secretion and bacterial clearance.

Characterization of hepcidin gene and protection of recombinant hepcidin supplemented in feed against Aeromonas hydrophila infection in Yellow River carp (Cyprinus carpio haematopterus).

Hepcidin is a small peptide of defensins with antibacterial activity, and plays an important role in innate immunity against pathogenic microorganisms, which can also participate in the regulation of iron metabolism. The hepcidin gene in Yellow River carp (Cyprinus carpio haematopterus) (CcHep) was cloned and identified. The total length of CcHep cDNA was 480 bp, containing an open reading frame (ORF) that encoded 91 amino acids (aa), which contained a 24-aa signal peptide, a 42-aa propeptide, and a 25-aa mature peptide. The mature peptide had a typical RX (K/R) R motif and eight conserved cysteine residues forming four pairs of disulfide bonds. Homology and phylogenetic tree analysis showed that CcHep had the closest relationship with that of crucian carp. The expression levels of hepcidin mRNA in healthy and Aeromonas hydrophila stimulated fish were measured by real-time fluorescence quantitative PCR. The results showed that CcHep mRNA was expressed in different tissues of healthy fish with the highest relative expression level in liver, followed by kidney and intestine, and the lowest expression level was observed in heart. The hepcidin gene was extremely significantly up-regulated in head kidney, intestine, liver, skin, spleen, and gill at 6 h and 12 h after A. hydrophila infection. Furthermore, the immunoregulation effect of dietary recombinant protein was evaluated. The recombinant hepcidin protein (rCcHep) was successfully expressed by Pichia pastoris X-33 and showed strong antibacterial activity against A. hydrophila, Escherichia coli, Vibrio anguillarum and Bacillus subtilis in vitro. In order to evaluate the preventive effect of rCcHep, fish were fed with basal diet or diet supplemented with different doses of rCcHep, and then challenged with A. hydrophila. The results showed that immune genes were up-regulated to varying degrees, and feed additive groups exhibited a significantly improved up-regulation expressions of Lysozyme, Toll-like receptor 5 (TLR 5), Major histocompatibility complex classⅡ (MHCⅡ), while inhibited up-regulation expressions of Interleukin 1ß (IL-1ß), Interleukin 8 (IL-8), and Tumor necrosis factor α (TNF-α) in liver and spleen compared to the control. Meanwhile, the relative immune protection rate in 120 mg/kg feed additive group was 28%, and the bacterial clearance rate in tissues of this group was higher than that of the control. Collectively, these results indicated that rCcHep had antibacterial activity and showed an immune protection effect against A. hydrophila, and could be considered as a dietary supplement to apply in aquaculture.

Supportive Care Needs Trajectories in Patients With Advanced Non-Small-Cell Lung Cancer Receiving Chemotherapy: A Longitudinal Study.

BACKGROUND: The supportive care needs trajectories in patients with advanced non-small-cell lung cancer (NSCLC) during chemotherapy and the related factors have yet to be explored or addressed in the literature. PURPOSE: This study was designed to investigate supportive care needs trajectories in patients with advanced NSCLC receiving chemotherapy and the association between the sociodemographic and disease characteristics of these patients over the four cycles of chemotherapy. METHODS: For this longitudinal study, 95 patients with advanced NSCLC were recruited using convenience sampling at a medical center in Taiwan. The supportive care needs of the participants were assessed in each of the four chemotherapy cycles using the Needs Evaluation Questionnaire-Chinese version (NEQ-C) with 23 dichotomous items on the day before and the seventh day after the end of each cycle. Group-based trajectory modeling was applied to identify the classes of supportive care needs trajectories, whereas chi-square tests were used to examine the factors related to these classes. RESULTS: Seventy-one participants completed all eight questionnaire sessions across the four cycles. The mean NEQ-C scores for these participants ranged between 14.4 and 14.6. Three classes of supportive care needs trajectories (low, moderate, and high) were identified for the entire NEQ-C and for each domain. Marital status was found to be associated with the classes of trajectories related to supportive care and assistance/care needs, spouse as the primary caregiver was found to be associated with the classes of trajectories related to information needs, and educational level was found to be associated with the classes of trajectories related to psychoemotional support needs. CONCLUSIONS: The results of this study indicate that marital status and spouse as primary caregiver relate significantly to supportive care needs trajectories in patients with advanced NSCLC during chemotherapy. Healthcare professionals should provide continuous, tailored supportive care interventions that address the needs of patients and their spouses/partners.

Molecular characteristics of interleukin (IL)-17A/F3 and its immune response on the pathogen and functional regulation on cytokines in common carp Cyprinus carpio L.

Fish interleukin (IL)-17A/F is homologous with mammalian IL-17A and IL-17F, which plays a key role in regulating inflammatory responses and autoimmune diseases. In fish, IL-17A/F1, 2, and 3 have been identified and described. However, IL-17A/F3 has received little attention in fish. In this study, a homolog of IL-17A/F3 was identified in common carp (Cyprinus carpio L.), which was termed as Cc_IL-17A/F3. The deduced amino acid sequence of Cc_IL-17A/F3 has four conserved cysteine residues, which could form two intrachain disulfide bonds. Homology comparison showed that the Cc_IL-17A/F3 was in the range of 31.7-71.9% of sequence similarity with these of other fishes. The Cc_IL-17A/F3 gene was constitutively expressed in various tissues, with higher expression levels in the skin and gills. After common carp were infected by Aeromonas. hydrophila, the mRNA expression levels of Cc_IL-17A/F3 were significantly up-regulated in the spleen, head kidney, gills, and intestine. Based on the indirect immunofluorescence assay, Cc_IL-17A/F3 proteins were found to be obviously increased in the intestine and spleen upon A. hydrophila infection at 24 h post-infection. The recombinant protein rCc_IL-17A/F3 could enhance the gene expression levels of pro-inflammatory cytokines (IL-1ß, IL-6, IFN-γ, and TNF-α) as well as chemokines (CXCL8 and CXCL20) in primary head kidney leukocytes. In vivo and in vitro experiments have similar stimulatory effects. When Cc_IL-17A/F3 was overexpressed in common carp, the expressions of pro-inflammatory cytokines and chemokines were significantly up-regulated in head kidney and spleen. In summary, the results derived from the present study suggested that the Cc_IL-17A/F3 plays an important role in defending against bacterial infections, and probably participates in mucosal immunity of the host.

Interleukin (IL)-22 in common carp (Cyprinus carpio L.): Immune modulation, antibacterial defense, and activation of the JAK-STAT signaling pathway.

Interleukin (IL)-22 is an IL-10 family cytokine secreted by CD4+ T cells and plays an important role in regulating inflammation and infection elimination. IL-22 homologues have been reported in the teleost, but the functions of IL-22 are still unclear. In this study, we identified two duplicated IL-22 genes in common carp (Cyprinus carpio L.), termed Cc_IL-22A and Cc_IL-22B. Sequence analysis showed that Cc_IL-22A and Cc_IL-22B had four conserved cysteine residues, which could form two intra-chain disulfide bridges. The Cc_IL-22A and Cc_IL-22B were constitutively expressed in various tissues, with the highest expression in the gill. The mRNA expression levels of Cc_IL-22A and Cc_IL-22B were significantly up-regulated in the gill, intestine, head kidney, and spleen of common carp challenged with Aeromonas. hydrophila. In vivo study showed that the expression levels of pro-inflammatory cytokines were significantly up-regulated in the head kidney and spleen when Cc_IL-22A or Cc_IL-22B were over-expressed. Furthermore, the over-expression of Cc_IL-22A and Cc_IL-22B indicated a protective effect on tissues, with only lymphocytic infiltration observed in comparison to the control and pcN3 groups, without obvious change in tissue morphology. Similar stimulatory effects of rIL-22A and rIL-22B were observed in vitro. When HKLs were stimulated with rIL-22A or rIL-22B, the expression levels of critical signaling molecules in the Janus kinase (JAK) and signal transducer and activator of transcription (STAT) pathway were significantly induced, including JAK1, JAK3, STAT1, and STAT3, as well as pro-inflammatory cytokines (IL-1ß and TNF-α). Together, these results suggest that Cc_IL-22A and Cc_IL-22B may regulate inflammatory responses through the JAK-STAT signaling pathway and have a significant impact on the immune defense of common carp against bacterial infection. Therefore, our study provides a new perspective on the functions of Cc_IL-22A and Cc_IL-22B in the immune defense mechanism of fish.

Significance of diagnosis and prognosis for appendiceal orifice inflammation in ulcerative colitis: a real-world study.

OBJECTIVE: Ulcerative colitis (UC) is a chronic inflammatory disease of the large intestine. At present, the significance of appendiceal orifice inflammation (AOI) in UC prognosis is still controversial. This prospective observational study investigated the importance of AOI in UC diagnosis and prognosis. Additionally, it compared the therapeutic efficacy of treatments in UC patients with or without AOI. DESIGN: This study was a prospective, observational, single-centre, real-world study. Patients with AOI were included in the observation group, and patients without AOI were assigned to the control group. All patients were followed up for 1 year; the disease remission and treatment efficacy were re-examined by colonoscopy. In addition, the clinical, endoscopic and pathological features were collected before and after the treatment. RESULTS: Patients with endoscopic diffuse inflammatory changes in the distal colorectum accompanied by AOI had a higher positive UC diagnosis rate than those without (96.5% vs 78.0%). Also, AOI had a specificity of 95.2% and a sensitivity of 28.3% for UC diagnosis. However, no difference in the modified Mayo score (p=0.881) or Baron grading was observed between the control and observation groups, indicating that AOI does not affect the treatment outcome of UC patients. CONCLUSION: In this study, the observation of AOI improved the UC diagnostic accuracy in patients with diffuse lesions in the distal colorectum. Furthermore, the presence of AOI does not affect the treatment efficacies of UC. TRIAL REGISTRATION NUMBER: ChiCTR1800017753.

Molecular characterizations, immune modulation, and antibacterial activity of interleukin-17A/F1a and interleukin-17A/F1b in common carp Cyprinus carpio.

Interleukin-17 (IL-17), as a pro-inflammatory cytokine family, mediates different pro-inflammatory mediators in various cell types (e.g., epithelial cells, macrophages, endothelial cells, and fibroblasts), which play an important role in defending against pathogens. The IL-17A/F1 genes have recently been reported in fish. However, the functions of these genes are still unclear. In this study, we identified two duplicated IL-17A/F1 genes in common carp (Cyprinus carpio L.), namely, CcIL-17A/F1a and CcIL-17A/F1b. Sequence analysis showed that CcIL-17A/F1a and CcIL-17A/F1b proteins had four conserved cysteine residues, which could form two intra-chain disulfide bridges. Homology comparison displayed that the deduced amino acid sequences of CcIL-17A/F1a and CcIL-17A/F1b shared 31.1%-77.3% and 32.5%-75.7% of sequence similarity to IL-17A/F1 homologues from other fish species, respectively. The mRNA expression levels of CcIL-17A/F1a and CcIL-17A/F1b were obviously increased in gill and head-kidney of fish challenged with A. hydrophila. The recombinant protein rCcIL-17A/F1a and rCcIL-17A/F1b could enhance the expression levels of pro-inflammatory cytokines (IL-1ß, IL-6, IFN-γ, and TNF-α) and chemokines (CXCL8 and CXCL20). The 3 × Flag eukaryotic expression vectors to express protein rCcIL-17A/F1a (or rCcIL-17A/F1b) were constructed and intramuscularly injected in common carp. The rCcIL-17A/F1a (or rCcIL-17A/F1b) could be successfully expressed in vivo. Four immune-related genes, namely, CD4, CD8, TNF-α, and IgM, were also significantly induced to be expressed at higher mRNA levels compared with the control. The pretreatment with CcIL-17A/F1a or CcIL-17A/F1b could markedly increase the survival rate of common carp challenged with A. hydrophila. Our results demonstrated that CcIL-17A/F1a or CcIL-17A/F1b plays an important role in immune responses and immune defense against bacteria. CcIL-17A/F1a or CcIL-17A/F1b could also be potentially used as an immunopotentiator to prevent diseases in common carp.

Selenium ameliorates mercuric chloride-induced brain damage through activating BDNF/TrKB/PI3K/AKT and inhibiting NF-κB signaling pathways.

Mercuric chloride (HgCl2), a heavy metal compound, causes neurotoxicity of animals and humans. Selenium (Se) antagonizes heavy metal-induced organ damage with the properties of anti-oxidation and anti-inflammation. Nevertheless, the molecular mechanism underlying the protective effects of sodium selenite (Na2SeO3) against HgCl2-induced neurotoxicity remains obscure. Therefore, the present study aimed to explore the protective mechanism of Na2SeO3 on HgCl2-induced brain damage in chickens. Morphological observations showed that Na2SeO3 alleviated HgCl2-induced brain tissues damage. The results also showed that Na2SeO3 decreased the protein expression of S100 calcium binding protein B (S100B), and increased the levels of nerve growth factors (NGF), doublecortin domain containing 2 (DCDC2), as well as neurotransmitter to reverse HgCl2-induced brain dysfunction. Further, Na2SeO3 attenuated HgCl2-induced oxidative stress by decreasing the level of malondialdehyde (MDA) and increasing the activities of total superoxide dismutase (T-SOD), glutathione peroxidase (GSH-Px), and total antioxidant capacity (T-AOC). Mechanistically, Na2SeO3 activated the brain-derived neurotrophic factor (BDNF)/tropomyosin-related kinase receptor type B (TrKB)/phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) signaling pathway and suppressed the nuclear factor kappa B (NF-κB) signaling pathway to inhibit apoptosis and inflammation caused by HgCl2 exposure. In summary, Na2SeO3 ameliorated HgCl2-induced brain injury via inhibiting apoptosis and inflammation through activating BDNF/TrKB/PI3K/AKT and suppressing NF-κB pathways.

Autophagy Delays Apoptotic Cell Death Induced by Siniperca chuatsi Rhabdovirus in Epithelioma Papulosum Cyprinid Cells.

Autophagy and apoptosis are two key cell fate determination pathways, which play vital roles in the interaction between viruses and host cells. Previous research had confirmed that one strain of fish rhabdoviruses, Siniperca chuatsi rhabdovirus (SCRV), could induce apoptosis and autophagy after infection. In the current study, we continued to analyze the interaction of autophagy and apoptosis in SCRV-infected EPC cell lines after treatment with different autophagy or apoptosis inhibitors. We found that SCRV infection could activate the mitochondrial apoptotic pathway by the detection of the activities of the caspase-3 and caspase-9 and by flow cytometry analysis in JC-1-stained cells, respectively. Furthermore, no significant autophagy-related factors were disturbed in SCRV-infected cell after apoptosis inhibitor Z-VAD-FMK treatment, while autophagy inducer rapamycin could obviously delay the occurrence of CPE and cell death. Meanwhile, rapamycin was able to reduce the proportion of apoptotic cells. Besides that, rapamycin could disturb the expression of p62 and LC3B-II, and the transcription level of SCRV nucleoprotein mRNA. The progeny virus titers did not show a big difference between the rapamycin treatment or without it. Collectively, our data preliminarily confirmed that SCRV-activated autophagy could delay apoptosis in EPC cells and may not affect virus production. Further study may need to focus on the crosstalk regulation and its roles on the SCRV infection.

Near infrared light fluorescence imaging-guided biomimetic nanoparticles of extracellular vesicles deliver indocyanine green and paclitaxel for hyperthermia combined with chemotherapy against glioma.

BACKGROUND: We investigated the therapeutic effect of targeting extracellular vesicles (EVs) loaded with indocyanine green (ICG) and paclitaxel (PTX) on glioma. METHODS: Raw264.7 cells were harvested to extract EVs for the preparation of ICG/PTX@RGE-EV by electroporation and click chemistry. We evaluated the success of modifying Neuropilin-1 targeting peptide (RGE) on the EV membrane of ICG/PTX@RGE-EV using super-resolution fluorescence microscopy and flow cytometry. Spectrophotometry and high performance liquid chromatography (HPLC) were implemented for qualitative and quantitative analysis of the ICG and PTX loaded in EVs. Photothermal properties of the vesicles were evaluated by exposing to 808-nm laser light. Western blot analysis, cell counting kit 8 (CCK-8), Calcein Acetoxymethyl Ester/propidium iodide (Calcein-AM/PI) staining, and flow cytometry were utilized for assessing effects of vesicle treatment on cellular behaviors. A nude mouse model bearing glioma was established to test the targeting ability and anti-tumor action of ICG/PTX@RGE-EV in vivo. RESULTS: Under exposure to 808-nm laser light, ICG/PTX@RGE-EV showed good photothermal properties and promotion of PTX release from EVs. ICG/PTX@RGE-EV effectively targeted U251 cells, with activation of the Caspase-3 pathway and elevated apoptosis in U251 cells through chemotherapy combined with hyperthermia. The anti-tumor function of ICG/PTX@RGE-EV was confirmed in the glioma mice via increased accumulation of PTX in the ICG/PTX@RGE-EV group and an increased median survival of 48 days in the ICG/PTX@RGE-EV group as compared to 25 days in the PBS group. CONCLUSION: ICG/PTX@RGE-EV might actively target glioma to repress tumor growth by accelerating glioma cell apoptosis through combined chemotherapy-hyperthermia.

Glioma exosomal microRNA-148a-3p promotes tumor angiogenesis through activating the EGFR/MAPK signaling pathway via inhibiting ERRFI1.

BACKGROUND: Glioma is the most frequent and lethal primary brain malignancy. Amounting evidence has highlighted the importance of exosomal microRNAs (miRNAs or miRs) in this malignancy. This study aimed to investigate the regulatory role of exosomal miR-148a-3p in glioma. METHODS: Bioinformatics analysis was firstly used to predict the target genes of miR-148a-3p. Exosomes were then extracted from normal human astrocytes and glioma cells. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was applied to determine the expression patterns of miR-148a-3p and ERBB receptor feedback inhibitor 1 (ERRFI1). Dual-luciferase reporter gene assay was applied to verify the direct binding between miR-148a-3p and ERRFI1. Cell counting kit-8 and tube formation assays were further conducted to assess the proliferation and angiogenic properties of human umbilical vein endothelial cells (HUVECs) in the co-culture system with exosomes. Lastly, glioma tumor models were established in BALB/c nude mice to study the role of exosomal miR-148a-3p in vivo. RESULTS: miR-148a-3p was highly expressed, while ERRFI1 was poorly expressed in glioma. miR-148a-3p was found to be enriched in glioma cells-derived exosomes and could be transferred to HUVECs via exosomes to promote their proliferation and angiogenesis. ERRFI1 was identified as a target gene of miR-148a-3p. In addition, miR-148a-3p activated the epidermal growth factor receptor (EGFR)/mitogen-activated protein kinase (MAPK) signaling pathway by inhibiting ERRFI1. In the co-culture system, our data demonstrated that glioma cells-derived exosomal miR-148a-3p down-regulated ERRFI1 and activated the EGFR/MAPK signaling pathway, so as to promote cell proliferation and angiogenesis. In vivo experimentation further demonstrated that this mechanism was responsible for the promotive role of exosomal miR-148a-3p in tumorigenesis and angiogenesis. CONCLUSION: Taken together, glioma-derived exosomal miR-148a-3p promoted tumor angiogenesis through activation of the EGFR/MAPK signaling pathway by ERRFI1 inhibition.

Identification of a protective epitope in Japanese encephalitis virus NS1 protein.

Japanese encephalitis virus (JEV) is one of the most important culex transmitted-flaviviruses, which can cause encephalitis in humans. Although non-structural protein 1 (NS1) of JEV does not stimulate neutralizing antibodies, this protein can provide high immunoprotection in vivo. The protective epitopes and the protective mechanism of NS1 still remain unclear. In this study, we generated five different monoclonal antibodies (mAbs) targeting the NS1 protein of JEV. In vitro experiments revealed that none of these five antibodies neutralized the JEV infection. In mouse protection studies, one of these mAbs, designated 2B8, provided a therapeutic effect against JEV lethal challenge (70% survival rate). Using peptide mapping analysis, we found that mAb 2B8 reacted with the epitope 225PETHTLWGD233 in the NS1 protein, in which any mutations among amino acid residues T228, H229, L231 or W232 could cause binding failure of 2B8 to the NS1 protein. Furthermore, mice immunized with KLH-polypeptide (225PETHTLWGD233) showed reduced mortality following JEV challenge. Collectively, we found a new protective epitope in the JEV NS1 protein. These results may facilitate the development of therapeutic agent and subunit-based vaccines based on the NS1 protein.

Emotional distress and quality of life during folinic acid, fluorouracil, and oxaliplatin in colorectal cancer patients with and without chemotherapy-induced peripheral neuropathy: A cross-sectional study.

When the 5-fluorouracil, leucovorin, and oxaliplatin (FOLFOX) chemotherapy regimen is used to treat colorectal cancer (CRC), chemotherapy-induced peripheral neuropathy (CIPN) caused by oxaliplatin can substantially affect quality of life (QOL) in the CRC patients. This study compared emotional distress and QOL during FOLFOX in CRC patients with and without CIPN symptoms.This cross-sectional, descriptive, and comparative study recruited 68 CRC patients receiving FOLFOX at a local teaching hospital and at a medical center in southern Taiwan. Self-reported structured questionnaires (oxaliplatin-associated neuropathy questionnaire, profile of mood states short form, and the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire, Core 30, version 3.0) were used for 1-time data collection. The Chi-square test, Fisher exact test, and Mann-Whitney U test were used to analyze data, and a P-value < .05 was considered statistically significant.The CIPN group had 45 (66.2%) patients, and the non-CIPN group had 23 (33.8%) patients. The 5 most common symptoms were coldness-related burning sensation or discomfort in the upper limbs, numbness in the upper limbs, tingling in the upper limbs, impairment of vision, and discomfort in the throat. The CIPN group had more females (P = .013), a more advanced stage of CRC (P = .04) and a higher chemotherapy dosage (P = .006). The 2 groups did not significantly differ in anxiety (P = .065) or depression (P = .135). Compared to the non-CIPN group, the CIPN group had significantly lower functioning (P = .001) and global health status (P < .001) and significantly more symptoms (P < .001).The CIPN group had significantly lower QOL compared to the non-CIPN group. However, the CIPN group did not have lower emotional distress compared to the non-CIPN group. The results of this study demonstrate the need for in-service courses specifically designed to train health professionals in assessing and managing CIPN symptoms to improve QOL in CRC patients receiving FOLFOX.

Associations among knowledge, attitudes, and practices toward palliative care consultation service in healthcare staffs: A cross-sectional study.

BACKGROUND: The palliative care consultation service (PCCS) of the National Health Insurance payments has been promoted in Taiwan since 2011, although few studies have been conducted on healthcare staffs' knowledge, attitudes, and practices regarding PCCS in Taiwan; consequently, the main objective of this study was to explore any correlations regarding the above by cross-sectional design using convenience sampling. METHODS: A total of 210 healthcare staff members were enrolled from a regional hospital from June 1, 2018, to September 30, 2018. Questionnaire items on the Palliative Care Consultation Service Inventory (KAP-PCCSI) were used to measure healthcare staff's knowledge, attitudes, and practices of PCCS. The collected data were analyzed by using descriptive statistics, independent samples t-test, Pearson's correlation coefficient analysis, and multiple linear regression analysis. RESULTS: The results revealed that the mean scores for knowledge of and attitudes of KAP-PCCSI were 58.7 ± 8.9 (perfect score: 75) and 42.7 ± 4.7 (perfect score: 50) respectively, while the mean score for practices of KAP-PCCSI was 36.3 ± 8.1 (perfect score: 50); moreover, the healthcare staff's knowledge and attitudes were positively correlated with their practices (p < 0.01). The results also showed that knowledge, attitudes, experience of having a family member(s) or friend(s) passing away, and being a medical personnel constituted the major predictors of practices (p < 0.001). These factors explained 43.2% of the overall variance for practices of KAP-PCCSI. CONCLUSIONS: The findings can help healthcare staff understand factors influencing practices of KAP-PCCSI and can serve as a reference for the development of strategies for palliative care education and training while improving the care quality of patients undergoing such palliative care with terminal life considerations in the hospitals, thereby fulfilling the goal of achieving holistic care.

Current Status and Changes in Pain and Activities of Daily Living in Elderly Patients with Osteoarthritis Before and After Unilateral Total Knee Replacement Surgery.

Knee osteoarthritis (OA) is a very common disease in the elderly, and total knee replacement (TKR) surgery is currently considered the most effective treatment. A prospective, observational, repeated measures study was performed to explore the current status and changes in pain and activities of daily living (ADL) in 58 OA elderly patients undergoing unilateral TKR. The Wong⁻Baker Faces Pain Rating Scale (WBS) for pain and the self-reported Barthel Index for ADL were measured on the day before surgery, 48 hours after surgery, and the day before discharge. Moderate pain was reported before surgery. Pain significantly improved after surgery and before discharge. At all three time points, pain scores were significantly higher in patients who used assistive devices compared to those who did not. Partial independence in ADL was reported before surgery. The ADL scores reported were highest before surgery, and those reported after surgery were lowest. However, ADL scores gradually increased before discharge. ADL scores were higher in the subjects who lived in a detached, single-family homes compared to those who lived in bungalows at all three time points. The results could be used to screen for knee OA elderly patients at high-risk for pain or low ADL and to provide timely intervention strategies as soon as possible.

Reduced nucleic acid methylation impairs meiotic maturation and developmental potency of pig oocytes.

Oocyte meiosis is a complex process coordinated by multiple endocrinal and molecular circuits. Recently, N6-methyladenosine (m6A) epigenetic modification on RNA is revealed to be important for meiotic maturation. However, the molecular mechanism of how m6A modification exerts its effect on oocyte maturation is largely unknown. Here, we showed that endogenous m6A writers (Mettl3 and Wtap) and eraser (Fto) elevated their transcript levels during meiotic maturation of pig oocytes. From germinal vesicle (GV) to metaphase II (MII) stages, global m6A level significantly increased, and existed mostly in ooplasm. Methyl donor (betaine, 16 mM) treatment of porcine cumulus-oocyte complexes (COCs) during in vitro maturation (IVM) significantly boosted nucleic acid m6A level within oocytes, but unchanged meiotic process and oocyte subsequent development. By contrast, methylation inhibitor (cycloleucine, 20 mM) reduced nucleic acid m6A level, and significantly decreased the germinal vesicle breakdown (GVBD) rate, the extrusion rate of the first polar body, and the cleavage and blastocyst rates of parthenotes. In addition, in cycloleucine-treated oocytes Wtap increased but Lin28 decreased their abundances significantly, along with the higher incidence of spindle defects and chromosome misalignment. Furthermore, pT161-CDK1 protein level in pig oocytes was confirmed to be decreased after cycloleucine treatment for 24 h. Taken together, chemical induced reduction of nucleic acid m6A methylation during pig oocyte meiosis could impair meiotic maturation and subsequent development potency, possibly through down-regulating pluripotency marker Lin28 mRNA abundance and disturbing MPF-regulated chromosome/spindle organization.