Indole-3-acetic acid ameliorates dextran sulfate sodium-induced colitis via the ERK signaling pathway.

Microbiota-derived catabolism of nutrients is closely related to ulcerative colitis (UC). The level of indole-3-acetic acid (IAA), a microbiota-dependent metabolite of tryptophan, was decreased significantly in the feces of UC patients. Thus supplementation with IAA could be a potential therapeutic method for ameliorating colitis. In this work, the protective effect of supplementation with IAA on dextran sulfate sodium (DSS)-induced colitis was evaluated, and the underlying mechanism was elucidated. The results indicated that the administration of IAA significantly relieved DSS-induced weight loss, reduced the disease activity index (DAI), restored colon length, alleviated intestinal injury, and improved the intestinal tight junction barrier. Furthermore, IAA inhibited intestinal inflammation by reducing the expression of proinflammatory cytokines and promoting the production of IL-10 and TGF-ß1. In addition, the ERK signaling pathway is an important mediator of various physiological processes including inflammatory responses and is closely associated with the expression of IL-10. Notably, IAA treatment induced the activation of extracellular signal-regulated kinase (ERK), which is involved in the progression of colitis, while the ERK inhibitor U0126 attenuated the beneficial effects of IAA. In summary, IAA could attenuate the clinical symptoms of colitis, and the ERK signaling pathway was involved in the underlying mechanism. Supplementation with IAA could be a potential option for preventing or ameliorating UC.

Tannic acid and carboxymethyl chitosan-based multi-functional double-layered hydrogel with pH-stimulated response behavior for smart real-time infection monitoring and wound treatment.

The dramatic increase of drug-resistant pathogenic bacteria has seriously effect on human health, appealing the needs of developing theranostic platforms with stimuli-responsive materials to realize the accurate bacterial diagnostics and therapeutics. Herein, a tannic acid and carboxymethyl chitosan-based multifunctional ZIF-90@i-PPOPs-phenol red double-layered hydrogel with stimuli-responsiveness and antibacterial activity was fabricated. The inner layer hydrogel (ZIF-90@i-PPOPs-based TFC hydrogels) was fabricated based on ZIF-90@i-PPOPs, integrate tannic acid and carboxymethyl chitosan linked by formylphenylboronic acid (FPBA), which exhibited outstanding injectable, biodegradability and antibacterial activity. The outer layer hydrogel (PR@PAM hydrogels) were constructed from polyacrylamide (PAM) and pH indicator phenol red, owning porous structure and excellent tissue adhesion. Due to the weakly acidic microenvironment within wound, the inner-layer hydrogel was stimulus-responsively decomposed, resulting in the accurate delivery of the positively charged ZIF-90@i-PPOPs to the lesion site to capture and kill bacteria by enhanced Zn2+ and ROS release. Meantime, the outer-layer hydrogel could real-timely monitor the pH changes to evaluate the wound recovery status. These double-layered hydrogels possessed precisely pH monitoring capacity, excellent antibacterial ability and negligible side effect to normal tissue in vivo, implying the high potential of the suggested hydrogels as theranostic platform for antibacterial treatment.

Isotope-Coded On-Tissue Derivatization for Quantitative Mass Spectrometry Imaging of Short-Chain Fatty Acids in Biological Tissues.

Short-chain fatty acids (SCFAs), as the main metabolites of gut microbiota, are recognized as crucial players in the host's inflammatory response and metabolic disease. Imaging the spatial distributions and calculating the accurate contents of SCFAs in the heterogeneous intestinal tissue are critical to reveal their biological functions. Here, we develop an isotope-coded on-tissue derivatization method combined with matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) to map the spatial expressions of SCFAs in the colon tissue based on pair-labeled N,N,N-trimethyl-2-(piperazin-1-yl)ethan-1-aminium iodide (TMPA) and D3-TMPA. A noticeable increase in the MALDI-MSI sensitivity of SCFAs was achieved after on-tissue derivatization, which enables the visualization of acetic acid, propionic acid, butyric acid, valeric acid, hexanoic acid, hydroxy acetic acid, and hydroxy propionic acid in the colon tissue. Moreover, the introduction of D3-TMPA-tagged SCFAs as internal standards can significantly reduce quantitation deviation from the matrix effects, ensuring the quantitative MALDI-MSI of SCFAs. We further used this method to characterize the spatial alterations of SCFAs in the colon tissues of mice with enterocolitis. The development of this strategy provides a reliable approach to image the spatial expressions of SCFAs in tissues and paves an insight way to study the roles of SCFAs in the gut microbiota and disease.

Tongxinluo capsule in combination with conventional therapies for stable angina pectoris: A meta-analysis and systematic review.

BACKGROUND: This study aimed to assess the efficacy and safety of Tongxinluo capsule (TXLC) in combination with conventional therapies for treating stable angina pectoris (SAP) through a comprehensive meta-analysis and systematic review. METHODS: We conducted a systematic search of the China National Knowledge Infrastructure, Wanfang, VIP, PubMed, Embase, and CENTRAL databases for randomized controlled trials investigating the use of TXLC as adjuvant therapy for SAP published up to June 2023. The Cochrane Handbook was used to evaluate the risk of bias. Meta-analysis was performed using Review Manager 5.4.1, and publication bias was assessed using Begg test and Egger test in the Stata SE 12.0 software. GRADEpro was used to assess the quality of the evidence. RESULTS: This meta-analysis included 26 randomized controlled trials with a total of 2352 patients. TXLC co-administration demonstrated significant reduction in angina attack frequency (mean difference (MD) -0.91, 95% confidence interval (CI) -0.97 to -0.84, P < .00001) and duration (MD -1.71, 95% CI -2.24 to -1.19, P < .00001), decreased use of nitroglycerin tablets (MD -6.28, 95% CI -7.16 to -5.41, P < .00001), lowered C-reactive protein (MD -1.19, 95% CI -1.35 to -1.03, P < .00001) and low-density lipoprotein cholesterol levels (MD -0.68, 95% CI -0.86 to -0.51, P < .00001). TXLC co-administration did not increase gastrointestinal reactions (RR 1.17, 95% CI 0.38 to 3.57, P = .78). The Begg test and Egger test results indicated no publication bias. The evidence quality was rated as very low for frequency of angina attack, duration of angina attack, and nitroglycerin usage, and low for C-reactive protein, low-density lipoprotein cholesterol levels, and gastrointestinal reaction events. CONCLUSION: This meta-analysis supports TXLC as a beneficial adjunct treatment for SAP.

End binding-3 inhibitor activates regenerative program in age-related macular degeneration.

Wet age-related macular degeneration (AMD), characterized by leaky neovessels emanating from the choroid, is a main cause of blindness. As current treatments for wet AMD require regular intravitreal injections of anti-vascular endothelial growth factor (VEGF) biologics, there is a need for the development of less invasive treatments. Here, we designed an allosteric inhibitor of end binding-3 (EB3) protein, termed EBIN, which reduces the effects of environmental stresses on endothelial cells by limiting pathological calcium signaling. Delivery of EBIN via eye drops in mouse and non-human primate (NHP) models of wet AMD prevents both neovascular leakage and choroidal neovascularization. EBIN reverses the epigenetic changes induced by environmental stresses, allowing an activation of a regenerative program within metabolic-active endothelial cells comprising choroidal neovascularization (CNV) lesions. These results suggest the therapeutic potential of EBIN in preventing the degenerative processes underlying wet AMD.

Construction of palladium porphyrins and triptycene photo-activated nanomaterial for enhanced colorimetric detection and inactivation of bacteria.

In the face of increasing bacterial resistance, design of high-performing and dual-functional nanomaterials to satisfy the requirements for both detecting and eradicating bacteria is of immense importance, but still remains a great challenge. Herein, a hierarchically three-dimensional (3D) porous organic frameworks (PdPPOPHBTT) was rationally designed and fabricated for the first time to realize ideal simultaneous detection and eradication of bacteria. PdPPOPHBTT covalently integrated palladium 5,10,15,20-tetrakis-(4'-bromophenyl) porphyrin (PdTBrPP, an excellent photosensitizer) with 2,3,6,7,12,13-hexabromotriptycene (HBTT, a 3D building module). The resulting material had outstanding NIR absorption, narrow bad gap and robust singlet oxygen (1O2) production capacity, which is responsible for the sensitive detection and effective removal of bacteria. We successfully realized the colorimetric detection of S. aureus and the efficient removal of S. aureus and E. coli. The first-principles calculations found at the highly activated 1O2 derived from the 3D conjugated periodic structures and ample palladium adsorption site in PdPPOPHBTT. The bacterial infection wound model revealed that PdPPOPHBTT possesses good disinfection ability and negligible side effect to normal tissue in vivo. This finding provides an innovative strategy for designing individual porous organic polymer (POPs) with multi-function and also broaden the applications of POPs as powerful nonantibiotic type of antimicrobials.

An Igh distal enhancer modulates antigen receptor diversity by determining locus conformation.

The mouse Igh locus is organized into a developmentally regulated topologically associated domain (TAD) that is divided into subTADs. Here we identify a series of distal VH enhancers (EVHs) that collaborate to configure the locus. EVHs engage in a network of long-range interactions that interconnect the subTADs and the recombination center at the DHJH gene cluster. Deletion of EVH1 reduces V gene rearrangement in its vicinity and alters discrete chromatin loops and higher order locus conformation. Reduction in the rearrangement of the VH11 gene used in anti-PtC responses is a likely cause of the observed reduced splenic B1 B cell compartment. EVH1 appears to block long-range loop extrusion that in turn contributes to locus contraction and determines the proximity of distant VH genes to the recombination center. EVH1 is a critical architectural and regulatory element that coordinates chromatin conformational states that favor V(D)J rearrangement.

Baicalein alleviates non-alcoholic fatty liver disease in mice by ameliorating intestinal barrier dysfunction.

Non-alcoholic fatty liver disease (NAFLD) is the main cause of chronic liver disease, and its pathological development is closely related to the gut-liver axis. The intestinal barrier, an important component of the gut-liver axis, can prevent gut microbes and endotoxins from entering the liver. Intestinal barrier function is impaired in patients with NAFLD. Baicalein, which is the main flavonoid in Scutellariae Radix, can improve NAFLD. However, whether baicalein alleviates NAFLD by ameliorating intestinal barrier dysfunction remains unclear. In this study, a methionine-choline deficient (MCD) diet-induced NAFLD mouse model is used. The effects of baicalein on lipid accumulation, inflammation and the intestinal barrier in MCD-fed mice were evaluated by detecting blood lipid levels, lipid accumulation, liver pathological changes, inflammatory factors, inflammatory signaling pathways, the three main short-chain fatty acids (acetate, propionate and butyrate), intestinal permeability and intestinal tight junction protein expression. Compared with the MCD-only group, baicalein intake decreased the serum and liver lipid levels. Moreover, the accumulation of lipid droplets and steatosis in the liver were also alleviated; all these results demonstrated that baicalein could alleviate NAFLD. Meanwhile, the levels of inflammatory cytokines decreased in the baicalein group. Further investigation of the mucosal permeability to 4 kDa fluorescein isothiocyanate-dextran, concentrations of short-chain fatty acids in feces, and the expression of intestinal zonula occluden 1 and claudin-1 indicated that a baicalein diet could decrease the intestinal permeability caused by a MCD diet. Moreover, the protein levels of p-NF-κB p65 and the ratio of p-NF-κB p65/NF-κB p65 increased, and IκB-α and PPARα decreased in NAFLD mice, while the administration of baicalein could alleviate these changes. The above results indicated that the mechanism of baicalein in the alleviation of NAFLD lies in the regulation of the intestinal barrier.

A metalloporphyrin and hydantoin functionalized nanozyme with synergistically enhanced bacterial inhibition.

An elaborate design of multimodal antibacterial agents has been revealed to be a promising strategy to address bacterial resistance, originating from the abuse of antibiotics. In this work, we have developed a positively charged and porous material, FePPOPHydantoin, as a disinfectant via introducing 1,3-dibromo-5,5-dimethylhydantoin (Hydantoin) and porphyrin iron units into a polymer framework. The extended π conjugated networks of FePPOPHydantoin endowed the material with strong near-infrared (NIR) absorption, high density of surface catalytic active centers, superior stability, and reproducibility. FePPOPHydantoin exhibits high peroxidase mimetic and photo-Fenton activity, which can catalyze the biologically allowable maximum concentrations of hydrogen peroxide (100 µM) to produce a vast amount of hydroxyl radicals. Simultaneously, the effective electrostatic interaction between the positively charged FePPOPHydantoin and the negatively charged bacteria facilitates the binding of FePPOPHydantoin on the bacterial membrane, restricting bacteria within the destruction range of hydroxyl radicals and thus making the bacteria more vulnerable. Finally, further close contact between bacteria and Hydantoin units in FePPOPHydantoin gave the material an antibacterial efficiency of over 99.999%. Compared with chemical therapy, photo-Fenton therapy, or peroxidase catalytic therapy alone, FePPOPHydantoin had a noteworthy multi-amplified antibacterial efficiency. Furthermore, FePPOPHydantoin exhibited good biocompatibility and negligible cytotoxicity. The in vivo antibacterial therapy on the Staphylococcus aureus (S. aureus) infected mouse wound model clearly proved the effectiveness of FePPOPHydantoin for fighting bacterial infections. This work highlights opportunities for the design of nanozymes with enhanced bacteriostatic activity, providing a new avenue for the construction of novel antibiotics.

Indole acetylated high-amylose maize starch: Synthesis, characterization and application for amelioration of colitis.

Tryptophan metabolites such as indole-3-acetic acid (IAA) are critical for gut health, through their binding to the aryl hydrocarbon receptor (AhR), and may be useful for treatment of gastrointestinal diseases. Delivery of IAA to the colon is necessary, and one strategy is use of esterified starches which get digested in the colon by gut microbes. High amylose maize starch (HAMS) resists digestion in the upper gastrointestinal tract and is fermented by gut microbiota to release short-chain fatty acids (SCFAs), which are also beneficial to intestinal homeostasis. IAA esterified to HAMS (HAMSIAA) was synthesized with different degrees of substitution (DSs) by controlling the ratio of IAA vs HAMS. Successful incorporation of indole acetyl group was verified by NMR and FTIR spectra. XRD revealed that the crystalline type of HAMSIAA changed from B to V-type. SEM showed the destroyed surface of the starch granules. HAMSIAA with DS ~ 0.3 effectively increased IAA in the colon, to levels unachievable by oral IAA delivery. HAMSIAA increased pathways downstream of AhR activation, including CYP1A1 mRNA expression and IL-22 protein levels, and greatly improved DSS-induced colitis. HAMSIAA could serve as an ideal means for colon-targeted delivery of IAA and a promising nutraceutical for amelioration of inflammatory conditions.

Establishment of Quality Evaluation Method for Yinqiao Powder: A Herbal Formula against COVID-19 in China.

Yinqiao powder, with significant anti-inflammatory and antiviral effects, is a classical formula for the treatment of febrile diseases in China. During the SARS period in 2003, Yinqiao powder showed a good antipyretic effect. It also plays a major role in the treatment for COVID-19 in China. Although there are many studies on the chemical compositions and pharmacological effects of Yinqiao powder, there are few studies on the quality standard system of it. In our study, a systematic quality evaluation method of Yinqiao powder combining HPLC fingerprint with quantitative analysis of multi-components by single marker (QAMS) based on network pharmacology and UPLC-Q-Exactive-Orbitrap-MS was established for the first time. In the UPLC-Q-Exactive-Orbitrap-MS experiment, a total of 53 compounds were identified in the extract solution of Yinqiao powder. In addition, 33 blood components were characterized, 23 of which were prototypes. The results of network pharmacology analysis showed that Yinqiao powder may inhibit inflammatory responses by suppressing IL-6, CXCL2, TNFα, NF-κB, etc., in the treatment of COVID-19. The HPLC fingerprint analysis of Yinqiao powder was conducted at 237 nm and 29 characteristic peaks were matched, 11 of which were identified. Forsythoside A was selected as the internal standard reference and double-wavelength (237 nm and 327 nm) was established in QAMS experiment. The repeatability was well under different conditions, and the results measured by QAMS were consisted with that of the external standard method (ESM), indicating that the QAMS method was reliable and accurate. The quality evaluation method of Yinqiao powder would be helpful to evaluate the intrinsic quality of Yinqiao powder more comprehensively, which is conducive to improve the quality standard of Yinqiao powder and provide a beneficial guarantee for the clinical treatment of COVID-19.

pH and Proton Sensor GPR65 Determine Susceptibility to Atopic Dermatitis.

pH sensing by GPR65 regulates various inflammatory conditions, but its role in skin remains unknown. In this study, we performed a phenome-wide association study and report that the T allele of GPR65-intronic single-nucleotide polymorphism rs8005161, which reduces GPR65 signaling, showed a significant association with atopic dermatitis, in addition to inflammatory bowel diseases and asthma, as previously reported. Consistent with this genetic association in humans, we show that deficiency of GPR65 in mice resulted in markedly exacerbated disease in the MC903 experimental model of atopic dermatitis. Deficiency of GPR65 also increased neutrophil migration in vitro. Moreover, GPR65 deficiency in mice resulted in higher expression of the inflammatory cytokine TNF-α by T cells. In humans, CD4+ T cells from rs8005161 heterozygous individuals expressed higher levels of TNF-α after PMA/ionomycin stimulation, particularly under pH 6 conditions. pH sensing by GPR65 appears to be important for regulating the pathogenesis of atopic dermatitis.

Promising Therapeutic Candidate for Myocardial Ischemia/Reperfusion Injury: What Are the Possible Mechanisms and Roles of Phytochemicals?

Percutaneous coronary intervention (PCI) is one of the most effective reperfusion strategies for acute myocardial infarction (AMI) despite myocardial ischemia/reperfusion (I/R) injury, causing one of the causes of most cardiomyocyte injuries and deaths. The pathological processes of myocardial I/R injury include apoptosis, autophagy, and irreversible cell death caused by calcium overload, oxidative stress, and inflammation. Eventually, myocardial I/R injury causes a spike of further cardiomyocyte injury that contributes to final infarct size (IS) and bound with hospitalization of heart failure as well as all-cause mortality within the following 12 months. Therefore, the addition of adjuvant intervention to improve myocardial salvage and cardiac function calls for further investigation. Phytochemicals are non-nutritive bioactive secondary compounds abundantly found in Chinese herbal medicine. Great effort has been put into phytochemicals because they are often in line with the expectations to improve myocardial I/R injury without compromising the clinical efficacy or to even produce synergy. We summarized the previous efforts, briefly outlined the mechanism of myocardial I/R injury, and focused on exploring the cardioprotective effects and potential mechanisms of all phytochemical types that have been investigated under myocardial I/R injury. Phytochemicals deserve to be utilized as promising therapeutic candidates for further development and research on combating myocardial I/R injury. Nevertheless, more studies are needed to provide a better understanding of the mechanism of myocardial I/R injury treatment using phytochemicals and possible side effects associated with this approach.

NMR resonance assignment and structure prediction of the C-terminal domain of the microtubule end-binding protein 3.

End-binding proteins (EBs) associate with the growing microtubule plus ends to regulate microtubule dynamics as well as the interaction with intracellular structures. EB3 contributes to pathological vascular leakage through interacting with the inositol 1,4,5-trisphosphate receptor 3 (IP3R3), a calcium channel located at the endoplasmic reticulum membrane. The C-terminal domain of EB3 (residues 200-281) is functionally important for this interaction because it contains the effector binding sites, a prerequisite for EB3 activity and specificity. Structural data for this domain is limited. Here, we report the backbone chemical shift assignments for the human EB3 C-terminal domain and computationally explore its EB3 conformations. Backbone assignments, along with computational models, will allow future investigation of EB3 structural dynamics, interactions with effectors, and will facilitate the development of novel EB3 inhibitors.

Potential of myricetin to restore the immune balance in dextran sulfate sodium-induced acute murine ulcerative colitis.

OBJECTIVES: Myricetin is a bioactive compound in many edible plants with anti-inflammatory and anticarcinogenic activity. The current study aimed to determine the protective effects and mechanism of myricetin against ulcerative colitis (UC). METHODS: Myricetin was orally administered at doses of 40 and 80 mg/kg to C57BL/6 mice with UC induced using dextran sulfate sodium. The disease-associated index and colon length were determined at the end of the experiment, the proportion of Treg, Th1 and Th17 was analysed by cytometry, and cytokines were detected using ELISA. KEY FINDINGS: Myricetin (80 mg/kg) ameliorated the severity of inflammation in acute UC and significantly improved the condition. Myricetin (80 mg/kg) elevated the levels of IL-10 and transforming growth factor ß. In addition, the proportion of regulatory T cells significantly increased in mice in the myricetin treatment group. CONCLUSIONS: Taking together, these results suggest that myricetin exhibits significant protective effects against UC and it could be used as a potential treatment for UC.

Amentoflavone protects the hematopoietic system of mice against γ-irradiation.

Some flavonoids have been shown to exhibit good antioxidant activity and protect mice from damage induced by radiation. Amentoflavone (AMF), a biflavonoid derived from the traditional herb-Selaginella tamariscina, has been reported to have antioxidant properties. The protective effects and mechanism of action of AMF against radiation injury remain unknown. In this study, male C57BL/6 mice were subjected to total-body 60Co γ-irradiation at 7.5 or 3.0 Gy. The survival rate and mean survival time were evaluated to determine the radioprotective effect of AMF. Number of peripheral blood cells, frequency of colony forming unit-granulocytes, monocytes and micronuclei were measured to assess the protective effects of AMF on the hematopoietic system. Levels of superoxide dismutase and glutathione, and pathological changes in the bone marrow were determined. Additionally, next-generation sequencing technology was used to explore potential targets of AMF. We observed that AMF markedly extends average survival time, reduces injury to the hematopoietic system and promotes its recovery. Furthermore, treatment with AMF significantly attenuated radiation-induced oxidative stress. In addition, AMF had a significant effect on gene tumor necrosis factor alpha-induced protein 2. Together, the results of this study suggest that AMF is a potential protective agent against radiation injury.

Berberine Protects Mice Against Dextran Sulfate Sodium-Induced Colitis by Activating mTORC1 Pathway.

Berberine is a plant alkaloid that can be extracted from many Chinese herbs. It has been reported that berberine could protect mice from ulcerative colitis, but the mechanism remains unclear. The current study's aim was to determine the potential mechanism by which berberine exhibits its anti-inflammatory function. Mice with colitis induced by dextran sulfate sodium (DSS) were administered with berberine at 50 mg/kg by gavage. Berberine significantly increased the proportion of regulatory T cells (Treg cells). The targeted metabolomics analysis was then performed to find that glutamine and glutamate metabolism played an important role in the process of regulating immune response. mTORC1 pathway was reported to closely relate with glutamine metabolism. As a result, the relative expression levels of downstream effector genes of mTORC were further determined, and data obtained showed that berberine could significantly increase the relative expression levels of S6K1 and 4EBP1. In addition, rapamycin was used to inhibit mTORC1 signaling, and it was found that colon length, disease associated index (DAI), and proportion of Treg cells of mice in the rapamycin-DSS group were not different from those of mice in the rapamycin/berberine-DSS group. Together, these results suggest that berberine exhibits significant protective effects against DSS colitis by activating the mTORC1 pathway to increase the proportion of Treg cells.

Sodium ferulate protects against influenza virus infection by activation of the TLR7/9-MyD88-IRF7 signaling pathway and inhibition of the NF-κB signaling pathway.

Influenza A virus (IAV) is highly contagious and causes considerable mortality worldwide. TLR3, 7, 8 and 9 recognize viral nucleic acids and rapidly trigger different signaling cascades that contribute to the production of interferons (IFNs) to antiviral defense. Therefore, a host immune response induced by the activation of these receptors can be used as a new antiviral strategy. In this study, the protective effect of sodium ferulate (SF) is investigated on mice infected with influenza virus A/FM/1/47(H1N1). SF improved survival and mitigated weight loss in infected mice. SF inhibited influenza virus replication by activating TLR7 and TLR9, which resulted in the promotion of IRF7 translocation into the nucleus and the production of typeⅠIFNs. Moreover, SF inhibited the NF-κB pathway by preventing p65 translocation from the cytoplasm to the nucleus. These findings demonstrate that SF plays a critical role in protection against IAV infection by activation of the TLR7/9-MyD88-IRF7 signaling pathway and inhibition of the NF-κB signaling pathway.

Antisense Oligonucleotides Targeting Raf-1 Block Japanese Encephalitis Virus In Vitro and In Vivo.

Japanese encephalitis virus (JEV) infections represent a major health concern in Southeast Asia since no effective treatments are available. Recently, several reports have demonstrated that inhibition of certain host cell proteins prevents viral infection. Raf-1 kinase is a central component of many signaling pathways involved in normal cell growth and oncogenic transformation, and Ras/Raf/ERK signaling activation has been observed during viral infections (including JEV infection). In this study, Raf-1 was confirmed to be upregulated by JEV infection, which suggested that Raf-1 might be important for JEV infection and might be a target for novel anti-JEV drugs. To determine the role of Raf-1 during the JEV infection process, antisense oligonucleotides (ASODNs) were used to downregulate Raf-1 expression in JEV-infected baby hamster kidney (BHK-21) cells and African green monkey kidney (Vero) cells. From five ASODNs candidates tested, Raf-1-1 (Raf-1 antisense) significantly downregulated Raf-1 protein expression levels, significantly inhibited cytopathic effect (CPE) in cultured cells, and reduced JEV RNA levels in cell medium without affecting cell viability. Furthermore, it also demonstrated that ASODN Raf-1-1 possessed therapeutic effects by using a lethal JEV infection mouse model. In conclusion, data presented in this report demonstrated that ASODN Raf-1-1 could suppress Raf-1 protein and that Raf-1 inhibition suppressed JEV replication in vitro and in vivo. These data provided evidence for targeting Raf-1 in the development of novel anti-JEV therapies. In addition, Raf-1-1 represents potential drugs that can be adapted for treating JEV infections.

Up-Regulated Expression of SPRY4-IT1 Predicts Poor Prognosis in Colorectal Cancer.

BACKGROUND Long non-coding RNA SPRY4 intronic transcript 1 (lncRNA SPRY4-IT1) has been reported to be associated with the progression of several cancers, but its expression level in colorectal cancer (CRC) has rarely been reported. The purpose of this study was to estimate the clinical significance of SPRY4-IT1 in CRC. MATERIAL AND METHODS The relative expression levels of SPRY4-IT1 were detected by quantitative real-time polymerase chain reaction (qRT-PCR) in diseased tissues and the adjacent normal tissues of 106 CRC patients. Chi-square method was used to evaluate the association between SPRY4-IT1 expression and the clinical features. Additionally, we assessed the overall survival at different expression levels of SPRY4-IT1 using Kaplan-Meier method. The prognostic significance of SPRY4-IT1 was estimated by Cox regression analysis. RESULTS Up-regulated level of SPRY4-IT1 was detected in pathologic tissues of CRC patients compared with adjacent normal tissues (P=0.000). The relative expression of SPRY4-IT1 was associated with the tumor size, the depth of invasion, lymph node invasion, distant invasion, and tumor stage (P<0.05). Patients with high expression of SPRY4-IT1 had poor overall survival compared with those with high level (39.3 vs. 49.3 months, log-rank test, P=0.016). Cox regression analysis showed that SPRY4-IT1 could act as an independent prognostic factor in CRC (HR=2.341, 95% CI=1.136-4.826, P=0.021). CONCLUSIONS SPRY4-IT1 might be associated with tumorigenesis and progression of CRC, and it may be a promising biomarker for prognosis in patients with CRC.