Higher temperatures due to climate change are causing greater sugar production in grapes and more alcoholic wines. The use of glucose oxidase (GOX) and catalase (CAT) in grape must is a biotechnological green strategy to produce reduced-alcohol wines. GOX and CAT were effectively co-immobilized by sol-gel entrapment in silica-calcium-alginate hydrogel capsules. The optimal co-immobilization conditions were achieved at a concentration of the colloidal silica, sodium silicate and sodium alginate of 7.38%, 0.49% and 1.51%, respectively, at pH 6.57. The formation of a porous silica-calcium-alginate structure was confirmed by environmental scanning electron microscopy and the elemental analysis of the hydrogel by X-ray spectroscopy. The immobilized GOX showed a Michaelis–Menten kinetic, while the immobilized CAT fits better to an allosteric model. Immobilization also conferred superior GOX activity at low pH and temperature. The capsules showed a good operational stability, as they could be reused for at least 8 cycles. A substantial reduction of 26.3 g/L of glucose was achieved with encapsulated enzymes, which corresponds to a decrease in potential alcoholic strength of must of about 1.5% vol. These results show that co-immobilized GOX and CAT in silica-calcium-alginate hydrogels is a promising strategy to produce reduced-alcohol wines. Global warming threatens to cause multiple complications due to the foreseeable increase in average temperatures in wine-growing areas and extreme meteorological  and biological (pests and diseases linked to fungi and/or bacteria) phenomena. These problems are already causing, with oscillations year after year, great uncertainty in the management of vineyards due to alterations in the vine and grape cycle that have repercussions on the normal and appropriate production of quality wines ,2. One o2. The amount of ethanol produced during fermentation is determined by the amount of sugar present in the must. High sugar and the resulting ethanol concentrations can lead to a number of microbiological, technological, sensory and financial problems [The phenology of the grapevine is driven by temperature. At higher average temperatures, a significant advance in the vegetative and reproductive cycles of the vine is triggered, anticipating the dates of budbreak, flowering and veraison. Thus, grape ripening (i) starts earlier, (ii) takes place at higher temperatures and (iii) the ripening season is lengthened to reach phenolic ripening. This leads to a higher sugar content and lower organic acid content in the grapes . As it iproblems . For exaproblems .This situation requires winegrowers to have technological alternatives according to the peculiarities of their crops and regions. Thus, on the one hand, adaptations in viticulture such as (i) modifications in plant material , (ii) vineyard management techniques  and (iii) site selection  are being tested to reduce the concentration of sugars in grapes . On the One of these strategies consists of a pre-fermentative treatment of the must with glucose oxidase (GOX) to reduce the concentration of glucose before the alcoholic fermentation, yielding wines with a lower alcoholic strength. At the same time, GOX has the potential to help balance the acidity of must with high pH due to gluconic acid production . Other p2O2) in the presence of oxygen (O2). It is common to use GOX in combination with catalase (CAT) to degrade H2O2 into O2 and H2O [2 also allows GOX to use it in a new glucose oxidation cycle. The low pH of the must (between 3.0 and 4.0) limits the glucose conversion by GOX. The relative enzyme activity of GOX from Aspergillus niger at pH 5.3 is 100%; however, it is drastically reduced by 50% at pH 3.3 [GOX is a flavoprotein that catalyzes the oxidation of glucose to gluconic acid and hydrogen peroxide  greater robustness and adaptability under process conditions, (ii) higher efficiency due to the reuse of the biocatalyst, (iii) the possibility of implementing continuous processes in the long term, (iv) better control of the catalysis by an easier separation of the enzyme from the must , (v) faster reaction rates and (vi) improved enzyme stabilization .Enzyme immobilization consists of the physical confinement of the enzyme in a region of space while preserving its enzyme activity. Several techniques have been developed for enzyme immobilization . EntrapmSeveral strategies have been explored to overcome the disadvantages of using only alginate gels and to improve their mechanical and chemical stability, increase their biocompatibility and control enzyme retention . One of To our knowledge, there are no studies focusing on the design and application of co-immobilized GOX and CAT to reduce glucose concentration in grape must, other than one applied on synthetic must with encapsulated GOX in alginate hollow beads . Our pap2 solution to obtain a hybrid interpenetrating polymer network of silica-calcium-alginate. We have reported that the hybrid gel prepared with this proposed coacervation process was stronger than the hybrid gel with a silica coating [In general terms, there are two strategies for the formation of hybrid gels of silica-alginate . The fir coating . Therefo2O2 degraded per g of capsules) determined in each type of capsule. Optimizing the immobilization conditions of an enzyme is critical since it is not possible to generalize its immobilization requirements . These rp ≤ 0.05). The analysis of variance determined that the selected quadratic model (Equation (1)) explained 95.9% of the variability of the data. A low coefficient of variation of the assays of the center point of the fractional design (runs 17–23) was obtained (8.35%), indicating a good precision of the experimental data  [At this optimal hydrogel composition, the porous structure of the gel could be ideal to (i) promote the transport of substrates and products, increasing the efficiency of the GOX catalysis and (ii) decrease the distance between GOX and CAT, increasing the degradation of Hperoxide , as wellperoxide , inducinperoxide . It has peroxide . It can 4.5–6.5) ,47.p ≤ 0.05) (2 (84.57%) was lower than the one observed for CG/cap though a similar value for the coefficient of variation of the center trials (9.31%) was achieved, also denoting a low experimental error. 2O2. As described for GOX, this effect could also be attributed to diffusion problems of substrate and/or unfavorable interactions between the enzymes and the immobilization support. Bower et al. [2O2 degraded per g of capsules and a predicted immobilization yield of 16.1%.Regarding CAT, the Pareto chart showed that only the concentration of colloidal silica was statistically significant ( ≤ 0.05) H. In ther et al.  reportedConsidering that the goal of this study was to optimize the conditions for the co-immobilization of GOX and CAT to maximize glucose consumption and that the variation of enzyme entrapment conditions resulted in greater changes for GC/cap than for HP/cap, we decided to perform the co-immobilization of both enzymes under the optimal conditions achieved for GOX.The morphological observations of calcium-alginate and silica-calcium-alginate capsules are shown in 2 in this capsule from colloidal silica and sodium silicate. The highest relative number of atoms was observed for the silicon atom (24.53%), indicating that the siliceous material was the main component of this capsule. The relative number of carbon atoms decreased from 44.94% in the calcium-alginate capsule to 15.72% in the silica-calcium-alginate capsule. On the contrary, the relative number of oxygen atoms increased from 33.47% to 51.39%, respectively. These changes in the relative number of carbon and oxygen atoms are due to the incorporation of colloidal silica and sodium silicate in the hydrogel of calcium alginate.The elemental analysis of calcium-alginate and silica-calcium-alginate capsules was carried out by X-ray spectroscopy. 2 = 0.99) (mK of the immobilized GOX (4.42 mM) was slightly higher than that of the free enzyme (3.88 mM), indicating that the hydrogel of silica-calcium-alginate partly limited the permeation rate of glucose through the porous capsule [mK. In contrast, the maxV of immobilized GOX (18.08 U) was smaller than that observed for free GOX (46.08 U), probably because of reduced glucose transport into the capsule. The values observed for mK and maxV suggest that the immobilization process partially reduced the GOX activity.To study the effect of co-immobilization on GOX kinetic, the initial reaction rates of free and immobilized GOX were measured at different glucose concentrations (1.0 to 27.0 mM) A. The ex = 0.99) B, sugges capsule . Rodrigu capsule  suggeste2O2 concentrations (3 to 16 mM)  of immobilized CAT was higher (2.60) than that of the free enzyme (2.24). Finally, a decrease in the maxV of CAT caused by the immobilization of the enzyme into the silica-calcium-alginate hydrogel was observed , probably due to the substrate diffusion problems into the capsule. The kinetic behavior of free and immobilized CAT was studied by evaluating the initial reaction rates of the enzyme at different Ho 16 mM) C. Free aTo determine the optimal pH values under oenological conditions for the co-immobilized GOX and CAT and compare them with the optimal pH of the free enzymes, the pH of the substrate solutions was changed between 3.0 and 4.0. Free GOX showed higher relative catalytic activity at pH values equal to or higher than 3.8 and decreased significantly in the lower pH range (3.0–3.4) , whereas2 was probably higher at lower temperatures since the solubility of this gas in an aqueous media increases as temperature decreases [2 production associated with this process, allowing the oxidative activity of GOX.Regarding the effect of temperature on GOX activity , the higecreases . The obsecreases , which aThe activity of the free CAT enzyme was quite similar over the range of temperatures tested. However, a higher activity for immobilized CAT was reached at 20 °C. In this case, the silica-calcium-alginate gel did not improve the CAT activity at low temperatures, unlike that observed for GOX at 10 °C.2O2, respectively, in 8 consecutive cycles of reusing the capsules. Although a higher relative activity of the immobilized GOX was obtained at 10 °C, we decided to evaluate its operational stability at the least favorable temperature of 25 °C. 2O2, which is a known GOX inhibitor [The success of any industrial enzyme application depends to a large extent on the maintenance of the enzyme activity during the operating time. The use of an immobilized enzyme with high operational stability reduces the cost of its application . The openhibitor ,58.Similar to our results, Ruiz et al. (2018)  reportedv/v) in the potential alcoholic strength of the must was achieved, estimating that 1.0% vol. (v/v) corresponds to a consumption of ~17 g/L of glucose by yeasts during alcoholic fermentation [The treatment of the must with the co-immobilized enzymes resulted in glucose consumption of 26.3 ± 0.2 g/L . Togetheentation . This reentation , which iAn effective immobilization support based on an interpenetrated polymer network of silica-sodium-alginate hydrogels was developed for the co-immobilization of GOX and CAT. This hydrogel showed a highly porous structure and enhanced GOX activity at low pH and temperatures. The control of the polymerization conditions was crucial for GOX and less so for CAT. The capsules showed a satisfactory behavior regarding their operational stability, as they could be reused for at least eight cycles. Moreover, the co-immobilization of GOX and CAT in silica-calcium-alginate hydrogels could be a very advantageous strategy to produce reduced-alcohol wines. A significantly decreased concentration of glucose in must was observed, which could reduce the excessive wine alcohol degree with a negative impact on its sensory quality and avoid the risk of stuck or sluggish fermentations. In addition, these immobilized enzymes could be very attractive for the biotechnological industry that demands new productive biocatalysts and new strategies to increase the useful life of enzymes. Further studies are required to test the technical feasibility of the silica-calcium-alginate immobilized GOX and CAT as a green strategy to reduce the alcoholic strength of the wine.® Mono 10.000 BG from A. niger, 10 KU/g) and catalase , were kindly provided by Novozymes . LUDOX® HS-40 colloidal silica (420816) and sodium silicate (338443) were purchased from Sigma-Aldrich . Sodium alginate (A3249), β-D-glucose (A1422) and H2O2   were purchased from Panreac Applichem . The rest of the chemicals were analytical quality grade and purchased from Panreac, S.A. .Glucose oxidase  contains a matrix of wheat flour that could interfere with the immobilization process. Therefore, a flour-free enzyme extract was prepared by dissolving 7.5 g of Gluzyme® Mono in 25.0 mL of 0.1 M citrate buffer at pH 6.6 with stirring at 225 rpm  for 30 min at 30 °C. The resulting solution was centrifuged at 2320× g for 15 min  and the supernatant was used as source of GOX (Solution 1). CAT was taken directly from the commercial enzyme preparation (Solution 2). The commercial GOX  g of glucose consumed per g of capsule (GC/cap) to assess the activity of the immobilized GOX and (ii) g of H2O2 degraded per g of capsule (HP/cap) to evaluate the activity of the immobilized CAT. On the one hand, colloidal silica and sodium silicate were dissolved together in distilled water at room temperature and the appropriate pH was adjusted with 1 N HCl (Solution 3). On the other hand, sodium alginate was dissolved at 40 °C in distilled water (pH previously adjusted with 1N HCl) (Solution 4). Solutions 3 and 4 were then mixed and kept under stirring at 1200 rpm for 3 min. 0.1 mL of Solution 1 (GOX) and 0.012 mL of Solution 2 (CAT) were added per mL of this homogenous mixture achieving a concentration of 117 U of GOX and 23 kU of CAT per mL of the final mixture. Stirring was maintained at 1200 rpm for 3 min until a complete homogenization. This final mixture was dripped with a 10 mL syringe  (at a height of 20 cm) into a 0.2 M CaCl2O2  dissolved in phosphate buffer  with 15 capsules for 2 min at 25 °C under agitation at 150 rpm. The initial and final H2O2 concentrations were measured by absorption spectroscopy at 240 nm [GC/cap was estimated by incubating 5 mL of β-D-glucose (20 mM) dissolved in sodium acetate buffer  with 15 capsules (~0.40 g) for 90 min at 25 °C under agitation at 150 rpm. The initial and final β-D-glucose concentrations were measured with an enzyme kit . HP/cap was estimated by incubating 10 mL of Ht 240 nm . 2O2, respectively, per min using the aforementioned assay conditions. The immobilization yield (in %) was calculated as ratio between the units of GOX or CAT activity immobilized to the units of GOX or CAT used initially for the immobilization.One unit of GOX or CAT activity was expressed as the amount of free or immobilized enzyme required to transform 1 mM of glucose or HWith the data of each assay for each dependent response variable, the optimized conditions were established using Statgraphics Centurion  fixing a second-order model for the independent variables with a significance level (α) of 0.05 and 15 coefficients as shown in Equation (1) .(1)y=βoThe structure of the capsules was analyzed by environmental scanning electron microscopy in a Quanta 200FEG ESEM . For this purpose, silica-calcium-alginate and calcium-alginate capsules with co-immobilized GOX and CAT were left to dry before analysis. These samples were examined in low vacuum (LV) mode with a large field electron detector. Additionally, the composition of the capsules was evaluated by environmental scanning electron microscopy coupled with energy-dispersive X-ray spectroscopy.2O2 in the range of 3 to 16 mM dissolved in phosphate buffer . Each assay was performed at 25 °C for 30 s. All assays were carried out under agitation at 150 rpm, in triplicate, and using 15 capsules for co-immobilized GOX and CAT, and the concentration of free enzymes corresponding to 15 capsules. mK is the Michaelis–Menten constant, The kinetic parameters for both the immobilized and the free GOX were calculated using 5 mL of β-D-glucose at different concentrations in the range of 1.0 to 27.0 mM dissolved in sodium acetate buffer . Each assay was performed at 25 °C for 5 min. The kinetic parameters for the immobilized and the free CAT were calculated using 10 mL of Htion (2) :(2)1VO=Kh is the hill slope. When h = 1.0, this equation is identical to the standard Michaelis–Menten equation. When it is bigger than 1.0, the curve is sigmoidal due to positive cooperativity. The calculations were performed with GRAPHPAD Prism 9 .The kinetic data for CAT showed a positive cooperative sigmoidal allosteric behavior of the enzyme. 2O2  dissolved in phosphate buffer (0.07 M), respectively, at six different pH values in the oenological range of 3.0 to 4.0. The effect of temperature was studied using four different temperature values in the range of 10 to 25 °C found in a winery. The other experimental conditions employed for the determination of GC/cap and HP/cap were used. For the free GOX and CAT, the concentration of free enzyme corresponding to 15 capsules was assayed. All pH and temperature assays were carried out in triplicate.The effect of pH on both free and co-immobilized GOX and CAT was assayed using β-D-glucose (20 mM) solution dissolved in sodium acetate buffer (0.1 M) or H2O2  in phosphate buffer  in 8 consecutive cycles of 90 min or 2 min each, respectively, with repeated use of the capsules. Fifteen capsules were used and washed after each cycle with 30 mL of the corresponding buffer for 30 s before starting the next cycle. Values were expressed as relative percentage of β-D-glucose for GOX and relative percentage of H2O2 for CAT. All assays were performed at 25 °C, shaking at 150 rpm, in triplicate.The operational stability of the co-immobilized GOX and CAT was determined by quantifying the glucose consumed in 5 mL of β-D-glucose (20 mM) in sodium acetate buffer  or 10 mL of Hg for 5 min, were treated with 15 capsules and maintained in an orbital shaking at 150 rpm for 48 h at 15 °C. A control must without capsules was maintained at the same conditions. After the treatment, the concentration of glucose and pH were measured in musts in triplicated. Three 50-mL Falcon tubes with 5 mL of grape must (191.9 g/L of glucose) at pH 3.8, centrifuged at 2320× p ≤ 0.05 using the Stratigraphic 19 Centurion statistical package .The analysis of the experimental design and variance of the experimental data were performed at a


A series of epoxy resins containing various trifluoromethyl groups were synthesized and thermally cured with diaminodiphenylmethane (DDM) and aminophenyl sulfone (DDS). All epoxy resins exhibited excellent thermal stability with the glass transition temperatures of above 128 °C and 5% weight loss temperatures of above 300 °C. DDS-cured epoxy resins possessed higher thermal stability than that of DDM-cured epoxy resins, while DDM-cured epoxy resins showed better mechanical, dielectric, and hydrophobic properties. Additionally, DDM-cured epoxy resins with different locations and numbers of trifluoromethyl groups showed flexural strength in the range of 95.55~152.36 MPa, flexural modulus in the range of 1.71~2.65 GPa, dielectric constant in the range of 2.55~3.05, and water absorption in the range of 0.49~0.95%. These results indicate that the incorporation of trifluoromethyl pendant groups into epoxy resins can be a valid strategy to improve the dielectric and hydrophobic performance. Epoxy resins have been widely applied in coatings, adhesives, advanced composites, and electronic materials ,2,3,4,5 It has been demonstrated that the modification on the backbone or side chains of polymers by introducing trifluoromethyl moieties has become one of the most efficient strategies to endow the materials with low dielectric constant, enhanced mechanical and thermal properties, as well as hydrophobic performance ,16,17. TIn this study, in order to figure out the relationship between the structure of fluorinated epoxy resins and their comprehensive properties as electronic packaging materials, a series of novel epoxy resins containing different positions and amounts of trifluoromethyl groups (FER) in the side chains were synthesized and then cured with aromatic amine 4, 4′-diaminodiphenylmethane (DDM) and 4, 4′-diaminophenyl sulfone (DDS). The curing kinetics of FER was studied by the iso-thermal DSC method. The thermal stability and mechanical and dielectric properties, together with water absorption, of fluorinated epoxy resins were investigated and compared in detail. The results show that the molecular structure of FER and the curing agent played a vital role in determining the performance and hydrophobic characteristics of cured epoxy resins. Moreover, this work provides a strategy to obtain alternative epoxy resins with a low dielectric constant and good moisture resistance by adjusting the position and amount of trifluoromethyl groups in the side chains. 3,5-Bis(trifluoromethyl)aniline (≥97.0%), 3-aminobenzotrifluoride (≥99.0%), 4-aminobenzotrifluoride (≥99.0%), and DDS (≥99.0%) were purchased from Energy Chemical, Shanghai, China. p-Benzoquinone (≥97.0%) was obtained from Aladdin chemistry Co., Ltd, Shanghai, China. Sodium nitrite, hydrochloric acid (37%), sodium hydrogen carbonate, toluene, epichlorohydrin, sodium hydroxide, acetone, and ethanol were all analytical pure and obtained from Beijing chemical company, Beijing, China. Zinc powder, tetrabutylammonium bromide, potassium acid phthalate, DDM, methyl red, and phenolphthalein were all analytical pure and purchased from Sinopharm Chemical Reagent Co., Ltd., Shanghai, China. All reagents were used as received without further purification.m-FPQ, p-FPQ, and d-FPQ containing various locations and numbers of trifluoromethyl substituents on the phenylhydroquinones were synthesized through diazo reaction and followed by a Meerwein arylation reaction, respectively. Aniline containing trifluoromethyl groups aniline, 0.1 mol), hydrochloric acid (33 mL), and deionized water (33 mL) were added into a beaker equipped with a mechanical stirrer and a thermometer. After cooling to 2 °C, sodium nitrite aqueous solution  was added dropwise into the beaker, and the temperature of the system was kept between 0 and 3 °C. The mixture was stirred for 30 min, and a transparent solution was obtained from filtration. Then the solution was added dropwise into a mixture of p-benzoquinone (0.08 mol), sodium hydrogen carbonate (0.03 mol), and deionized water (100 mL). The mixture was stirred at 8~10 °C for 4 h. After filtration, the precipitate was washed with deionized water. The obtained yellow solid was dried at room temperature.As shown in The above yellow solid (0.05 mol), zinc powder (0.15 mol), and deionized water (60 mL) were added into a three-neck flask equipped with a mechanical stirrer, a dropping funnel, and a condenser. The mixture was heated to 90 °C, and then hydrochloric acid (35 mL) was added dropwise into the flask. The reaction mixture was allowed to reflux for 4 h and then filtered. The precipitate was recrystallized from toluene and dried in a vacuum oven for 24 h.m-FPQ: yield: 80%, melting point: 88 °C. 1H-NMR  δ 9.04 , 8.90 , 7.92–7.73 , 7.72–7.53 , 6.79 , 6.72 , 6.65 . Elem. Anal. Calcd. for C13H9F3O2: C, 61.42%; H, 3.57%; O, 12.59%. Found C, 61.51%; H, 3.44%; O, 12.72%. p-FPQ: yield: 80%, melting point: 111 °C. 1H-NMR  δ 9.01 , 8.88 , 7.74 , 6.79 , 6.72 , 6.65 . Elem. Anal. Calcd. for C13H9F3O2: C, 61.42%; H, 3.57%; O, 12.59%. Found C, 61.34%; H, 3.56%; O, 12.60%.d-FPQ: yield: 75%, melting point: 112 °C. 1H-NMR  δ 9.15 , 8.85 , 8.08 , 7.92 , 6.73 , 6.61 . Elem. Anal. Calcd. for C14H8F6O2: C, 52.19%; H, 2.50%; O, 9.93%. Found C, 52.22%; H, 2.43%; O, 10.02%.m-FER, p-FER, and d-FER were prepared by a similar procedure. Taking m-FER as an example, the procedure is described as the following. A mixture of m-FPQ (0.1 mol), epichlorohydrin (157 mL), and tetrabutylammonium bromide (1.93 g) was mixed in a three-neck flask at 90 °C for 6 h. The excess epichlorohydrin was removed at a reduced pressure. Then, toluene (160 mL) and sodium hydroxide  were added to the mixture, and the reaction was kept at 90 °C for another 3 h. Then, the mixture was washed with deionized water at room temperature until the water was neutral. The toluene and water were distilled under reduced pressure. The product was dried in the oven at 80 °C for 24 h.According to the location and number of trifluoromethyl substituents on the FERs, m-FER: yield: 85%, liquid at room temperature, epoxy value: 0.52. 1H-NMR  δ 7.44 , 7.23 , 7.09–6.98 , 6.96–6.84 , 4.32 , 4.24 , 3.37–3.28 , 3.22 , 2.79 , 2.71–2.58 . Elem. Anal. Calcd. for C19H17F3O4: C, 62.29%; H, 4.68%; O, 17.47%. Found C, 61.88%; H, 4.70%; O, 17.39%.p-FER: yield: 87%, melting point: 115 °C, epoxy value: 0.52. 1H-NMR  δ 7.78 , 7.15–7.07 , 7.04–6.94 , 4.33 , 3.85 , 3.36–3.32 , 3.25 , 2.86–2.81 , 2.81–2.74 , 2.71 , 2.62 . Elem. Anal. Calcd. for C19H17F3O4: C, 62.29%; H, 4.68%; O, 17.47%. Found C, 62.03%; H, 4.58%; O, 17.35%.d-FER: yield: 80%, melting point: 110 °C, epoxy value: 0.44. 1H-NMR  δ 8.24 , 8.07 , 7.15 , 7.05 , 4.35 , 3.94–3.85 , 3.34 , 3.22 , 2.80 , 2.73–2.56 . Elem. Anal. Calcd. for C20H16F6O4: C, 55.31%; H, 3.71%; O, 14.73%. Found C, 55.29%; H, 3.70%; O, 14.68%.Ti), the peak curing temperature (Tp), and the end-set curing temperature (Te) of epoxy resins are listed in Ti, Tp, and Te [Ti for 2 h, at Tp for 2 h, and then at Te for 1 h.The epoxy resins were cured with two kinds of curing agents, DDM and DDS, respectively. According to the epoxy values, the stoichiometric amounts of DDM and DDS were added to the epoxy mixtures with acetone as the solvent. Then, the mixtures were stirred for 30 min at room temperature. The solution was poured into a mould. The mould was dried in a vacuum oven at 40 °C for 12 h to remove the solvent. The onset curing temperature  at 500 MHz using deuterated dimethyl sulfoxide (DMSO-d6) as the solvent and tetramethylsilane (TMS) as the standard. The elemental contents of C, H, and O were determined using an elemental analyzer . The epoxy value was determined via the acetone-hydrochloride method, and methyl red was used as the indicator. The epoxy value was calculated as the mean of the three tests. The cure kinetics and glass transition temperature (Tg) of the cured epoxy resins were measured by using DSC  under nitrogen at a flow rate of 50 mL/min. Thermogravimetry analysis (TGA) was performed on the Perkin-Elmer Pyris1 thermogravimetric analyzer  from 80 to 800 °C at a heating rate of 10 °C/min under a nitrogen atmosphere. The flexural properties of samples were evaluated at room temperature on a universal tensile testing machine  at a speed of 1 mm/min. The dielectric constant and dielectric loss of epoxy resins were measured at room temperature by a 4292 precision impedance analyzer  in a range of frequencies from 1 kHz to 1 MHz. Water absorption was measured by immersing epoxy resins in water at 80 °C for 48 h, which was then wiped with tissue paper and weighted to determine the water absorption value. The water contact angle was measured using a contact angle goniometer  at room temperature.1H-NMR and elemental analysis, as shown in m-FER, p-FER, and d-FER were 0.52, 0.52, and 0.44, which were close to the theoretical epoxy values. All these results indicate that FERs were synthesized successfully.The structure of FPQs and FERs was confirmed by m-FER: liquid; p-FER: 115 °C; d-FER: 110 °C). This phenomenon was also observed from the DSC curves of DDS-cured m-FER and p-FER with one endothermic peak. However, there are two endothermic peaks (110 and 180 °C) in the DSC curves of DDS-cured d-FER. The corresponding temperatures of endothermic peaks are in accordance with the independent melting points of the immiscible mixture (d-FER and DDS). Furthermore, there is only one exothermic peak in the curves for all samples. Similar to the published work [Tp) in their DSC curves shows an increasing tendency as the heating rate increases. Ti, Tp, and Te of DDM and DDS-cured fluorinated epoxy resins. Compared to DDM-cured FERs, all the Ti, Tp, and Te of DDS-cured samples are much higher. It can be attributed to the electron-withdrawing sulfone in DDS, which results in the weaker reactivity of DDS and higher curing temperature. Moreover, all the Ti, Tp, and Te of DDS-cured d-FER samples are higher than those of other kinds of FERs. This is because the introduction of two trifluoromethyl substituents on the phenyl side chain decreases the reactivity between FERs and DDS. The investigation of curing kinetics can be helpful to produce high-performance thermosetting polymers. The curing behaviors of FERs were investigated by the iso-thermal DSC method using DDM and DDS as the curing agents at different heating rates  . The dynhed work , the temEa) of FERs was determined by the Kissinger method [R is the ideal gas constant, 8.314 J/(mol·K). In this study, the overall activation energy  versus 1/pT, as displayed in aE of DDM-cured FERs are in a range of 41.1~53.9 kJ/mol, while the calculated aE of DDS-cured FERs are in a range of 58.8~66.6 kJ/mol. The higher aE of the DDS system implies that the DDS is a curing agent with weaker reactivity, which requires higher curing temperatures. Furthermore, the nth order of the curing reaction can be calculated from Crane’s equation [Therefore, the equation : (2)d(lnn) of 0.92 ± 0.02, indicating the curing reaction mechanism between FERs and curing agents (DDM and DDS). The curing process was accomplished by the epoxy ring-opening reaction between the epoxy group and the primary amino group of curing agents.It was found that all these curing epoxy systems possessed a similar reaction order . The incorporation of two trifluoromethyl substitutions may result in higher electrostatic repulsion than intermolecular attraction, thus increasing the polymer chain spacing and then leading to a slightly lower Tg.The 0, Wt, and W∞ are the initial weight, the weight at a given temperature, and the final weight of the sample recorded by TGA As shown in Td5 and Td10), as presented in 3 substitutions on the phenyl side chain, the cured d-FER samples exhibited higher Td5 and Td10 values than those of m-FER and p-FER samples. Moreover, DDS-cured epoxy resins possessed better thermal stability than DDM-cured samples, owing to the strong inter-chain interactions deriving from sulfone moiety with high polarization. As a result, the thermal stability of epoxy resins can be enhanced by introducing trifluoromethyl groups and increasing the inter-chain interactions.The thermal stability of DDM and DDS-cured FERs was evaluated by TGA. As shown in d-FER samples cured by DDM and DDS exhibited moderate flexural strengths (127.00 and 118.85 MPa). This was possibly attributed to the rigid biphenyl scaffold in the chains and two polar −CF3 groups in the side chain.To determine the mechanical flexibility of these cured FERs for electronic packaging application, we carried out a 3-point bending test on their rectangular-shaped samples and then obtained the flexural stress and modulus ,30. The k materials which are applied in microelectronics [d-FER samples exhibited much lower water absorption values than those of the others. Among them, DDM-cured d-FER with two −CF3 substitutions on the phenyl side chain possessed the lowest water absorption of 0.49% due to the low surface energy and hydrophobic characteristic of trifluoromethyl groups [It is crucial to possess low water absorption for low-ctronics . The watl groups . This wal groups . The resd-FER-DDM showed the highest water contact angle of 108°, implying that the incorporation of trifluoromethyl substitutions makes the materials more hydrophobic and prevents the moisture from being absorbed into epoxy resin [In order to investigate the nature of water absorption, the water contact angle of these cured FERs was measured, and the results are shown in xy resin . 3 substitutions possess much low polarization and a higher free volume [d-FER samples present the lowest dielectric constant due to the incorporation of a higher number of trifluoromethyl substitutions. Among them, the dielectric constant and dielectric loss of d-FER-DDM are as low as 2.55 and 0.016 at 1 MHz, respectively. These results indicate that these cured FERs have great potential for applications in electronic packaging and microelectronics.For the microelectronic packaging application, the alternative epoxy resin must possess a low dielectric constant, as well as low dielectric loss, to reduce the delay of signal transmission ,33. Figue volume . The curd-FER with two −CF3 substitutions on the phenyl side chain exhibited a large water contact angle of 108°, a low water absorption of 0.49%, a low dielectric constant of 2.55, and dielectric loss of 0.016 at 1 MHz. In summary, the incorporation of trifluoromethyl groups into the side chain of epoxy resins can improve the hydrophobic characteristics, moisture resistance, and dielectric properties of cured epoxy resins. Combing their good thermal and mechanical properties, epoxy resins containing pendent trifluoromethyl groups, especially DDM-cured d-FER, show great prospects in matrix materials and applications in microelectronics.A series of epoxy resins with various locations and numbers of trifluoromethyl substitutions were synthesized via a two-step reaction and thermally cured with aromatic amine DDM and DDS. The performance of cured epoxy resins was affected by the molecule structure of epoxy resins and curing agents. The obtained epoxy resins exhibited high glass transition temperatures as well as low water absorption and dielectric constant. The cured FERs also displayed good mechanical properties and high thermal stability. Especially, DDM-cured


Aberrant Wnt signaling activation is frequently observed in many cancers. The mutation acquisition of Wnt signaling leads to tumorigenesis, whereas the inhibition of Wnt signaling robustly suppresses tumor development in various in vivo models. Based on the excellent preclinical effect of targeting Wnt signaling, over the past 40 years, numerous Wnt-targeted therapies have been investigated for cancer treatment. However, Wnt signaling-targeting drugs are still not clinically available. A major obstacle to Wnt targeting is the concomitant side effects during treatment due to the pleiotropic role of Wnt signaling in development, tissue homeostasis, and stem cells. Additionally, the complexity of the Wnt signaling cascades across different cancer contexts hinders the development of optimized targeted therapies. Although the therapeutic targeting of Wnt signaling remains challenging, alternative strategies have been continuously developed alongside technological advances. In this review, we give an overview of current Wnt targeting strategies and discuss recent promising trials that have the potential to be clinically realized based on their mechanism of action. Furthermore, we highlight new waves of Wnt targeting that combine recently developed technologies such as PROTAC/molecular glue, antibody–drug conjugates (ADC), and anti-sense oligonucleotides (ASO), which may provide us with new opportunities to target ‘undruggable’ Wnt signaling. Over the four decades since its discovery, Wnt-mediated signaling has been extensively studied, revealing its evolutionarily conserved roles in regulating diverse cellular processes, including embryonic development, tissue homeostasis, and cell fate determination [In the early 1980s, Wnt signaling was first discovered when or virus . Throughmination .Wnt signaling consists of Wnt ligands, frizzled receptors (FZD family), co-receptors, β-catenin destruction complexes, β-catenin/transcriptional partners, and other modulating components . Wnt is Herein, we briefly describe Wnt signaling transduction, the understanding of which plays a key role in the development of targeting strategies .So far, 19 secreted Wnt ligands and more than 18 Wnt receptors/co-receptors have been identified in the mammalian system . DiffereThe β-catenin-dependent Wnt signaling pathway is more elucidated and established compared to the β-catenin-independent pathway . β-caten2+ , which releases Ca2+ from the intracellular stores. The raised levels of Ca2+ induce the activation of protein kinase C, protein kinase II, and calcineurin [In β-catenin-independent signaling, the intracellular downstream signaling of the Wnt/receptor interactions is mediated not by β-catenin but by various cellular signaling modules, such as RAC1-JNK, RHOA-ROCK, and PLC-IP3-Ca2+ . Well-chignaling . In Wnt/ignaling . Consequignaling . The Wntcineurin ,24. AlthAPC, ZNRF3, CTNNB1 (encodes β-catenin), AXIN1, and RSPOs [Aberrant Wnt signaling has been implicated in the tumorigenesis of various cancers . For exand RSPOs  , the loss-of-mutation of observed ,27. The ut 5~10% . The mutl tumors . In addipatients . Mutatioigenesis .APC mutations are detected [APC mutation is sufficient to initiate gastric tumors in in vivo models [RNF43, another Wnt signaling component, were observed in 4.3% of the MSS (microsatellite stable) subtype (~20% of GC patients) and 54.6% of the MSI (microsatellite instable) subtype (~80% of patients) and were considered key driver mutations [Wnt1 and Wnt6 has been observed in gastric cancer [Wnt1 or a combination of prostaglandin pathways induces preneoplastic lesions or invasive gastric cancer, respectively, in mouse models [CDH1 mutation (encodes E-cadherin) and Helicobacter pylori infections [In gastric cancer (GC), 13~22% detected ,27. APC utations ,30. In ac cancer ,32. The e models . Indeed,fections ,35,36. TCTNNB1, encoding β-catenin, ~10% of AXIN1, and ~3% of AXIN2, are mutated [CTNNB1, the most frequent mutation in HCC, does not induce a tumor, the loss-of-APC or Axin1 mutation, both less common in HCC, are able to do so [In hepatocellular carcinoma (HCC), almost 50% of mutations in the Wnt signaling pathway are activation mutations . Approxi mutated ,38. Addi mutated . These gto do so ,41.In addition to the oncogenic role of Wnt signaling, many genetic alterations in the Wnt signaling of specific types of cancers do not drive tumorigenesis but play a tumor-promoting role during cancer progression ,42. AddiCTNNB1 and APC genes is not common [APC and CHD8, which inhibits transcription mediated by CTNNB1, is related to the relapse of SCLC [In lung cancer, the mutation of t common ,46. Howet common ,47. In s of SCLC . In non- of SCLC . A recen of SCLC . MoreoveRNF43 and <1% mutations in APC and CTNNB1 [Pancreatic ductal acinar cell carcinoma (PDAC) exhibits approximately 4~10% mutations in d CTNNB1 . Althougd CTNNB1 ,50. Inded CTNNB1 ,51.Since aberrant Wnt activation initiates or promotes tumorigenesis, Wnt-targeting strategies in cancer therapy are directed toward the downregulation or restoration of overactivated Wnt signaling. Current Wnt targeting strategies can be classified into four categories according to the target location in Wnt signaling: targeting Wnt ligands, targeting Wnt receptors, targeting destruction complex, and targeting β-catenin/transcriptional factors. Extracellular Wnt ligands and receptors are good targets for specific antibodies. However, intracellular signaling components, such as the destruction complex and β-catenin/transcriptional factors, cannot be targeted with antibodies. Only small chemicals and peptides are available to target those. Thus, the enzymatic domains of Wnt signaling components are considered key targets of intracellular components.In this section, we describe current Wnt targeting strategies as well as recent updates, with meaningful clinical trials involving each strategy .Targeting cancer-specific Wnt ligands in certain cancers may be an excellent way to increase the specificity of cancer treatment. However, Wnt ligands often act redundantly, and which Wnt ligands are essential is not fully understood in each cancer context . AdditioIndeed, the PORCN inhibitor LGK974  prevents the secretion of Wnt by inhibiting the palmitoylation of Wnt and shows dramatic suppression of lung tumorigenesis and metastatic CRC progression ,54. In mSoluble Wnt modulators could also be a good target for Wnt-targeted therapy. Soluble FZD-related proteins (SFRP) restrain the Wnt signaling pathway. SFRP has a cysteine-rich domain (CRD) homologous to FZD-CRD. This domain acts as an SFRP to bind competitively to Wnt ligands in order to inhibit Wnt signaling ,130. V3NThe targeting of another soluble Wnt modulator, dickkopf-related protein family (DKK), has proved to be more promising in clinical trials. DKKs are composed of five types, of which dickkopf-1, DKK1, binds to LRP5/6 and was initially considered to inhibit canonical Wnt signaling ,19,131. Targeting Wnt receptors is challenging in cancers harboring mutations downstream of Wnt signaling components. However, cell surface receptors are relatively easy to develop antibodies for, making them attractive drug targets. Additionally, antibodies targeting cancer-specific Wnt receptors could give more specificity with fewer side effects . FurtherA fair number of FZD-targeting antibodies are currently undergoing clinical trials . A monocNaturally secreted R-spondins induce the accumulation of FZD receptors and enhance Wnt signaling. Thus, inhibiting natural or cancer-specific RSPOs could be a therapeutic target for Wnt signaling. OMP-131R10 (Rosmantuzumab) is a monoclonal antibody targeting R-spondin 3 (RSPO3) that has now completed a phase 1 clinical trial in advanced solid or relapsed tumors (NCT02482441)  Table 1.The β-catenin destruction complex, consisting of APC, CK1, AXIN, and GSK3β, plays a vital role in controlling the concentration of cytosol and nuclear β-catenin . Loss-ofCK1 and GSK3β phosphorylate β-catenin, which subsequently induces the ubiquitination and proteasomal degradation of β-catenin . Thus, aSulindac is an FDA-approved non-steroidal anti-inflammatory drug . Later, Another way to enhance the destruction of complex-mediated β-catenin degradation is to stabilize the AXIN. Tankyrase belongs to the poly ADP-ribose polymerase family and induces the proteasomal degradation of AXIN ,147. Thu+-ATPase (v-ATPase) mediates the vesicular acidification of lysosomes. This acidification of lysosomes is required for the lysosomal degradation of Wnt receptors/co-receptors and APC, which hyperactivates Wnt/β-catenin signaling [Hignaling ,149. Thuignaling . A substAPC mutations, enhancing the destruction complex’s function is impossible. In addition, enhancing destruction complexes via v-ATPase inhibitors can lead to a broad range of potential side effects induced by pan-lysosomal inhibition.Targeting the intracellular protein complex is quite challenging. Moreover, since most Wnt-associated cancers carry β-catenin acts as a central transcriptional activator by forming a complex with its transcriptional partner, TCF/LEF. β-catenin also recruits co-activators such as cAMP response element binding protein (CBP) and p300 ,21. The ICG-001 and its analog PRI-724 interfere with the interaction between β-catenin and CBP ,81. IndeTheoretically, interfering with the binding of β-catenin to its transcriptional partner might be an excellent strategy. It would also be ideal if we could specifically inhibit the mutant beta-catenin. However, it is tremendously difficult to understand the specific interactions between proteins. Moreover, finding chemicals that prevent such interactions is another obstacle. Although there are many limitations, several small-molecule compounds are undergoing pre- or clinical tests , and newDespite ongoing clinical trials, devastating adverse effects on tissue homeostasis and regeneration occur while targeting Wnt signaling. Therefore, Wnt signaling-targeted therapy holds excellent promise but carries high risks in cancer treatment . In thisAPC or CTNNB1 mutations downstream of Wnt ligands and receptors. Similarly, targeting intracellular destruction complexes is also not very beneficial for these Wnt mutant cancers.As has been mentioned elsewhere, targeting Wnt ligands and receptors against antibodies can give specificity to cancer therapy and cause fewer side effects compared to chemical drugs . HoweverAlthough targeting mutant or nuclear β-catenin is theoretically ideal, finding small chemical inhibitors that prevent β-catenin interactions is extremely challenging.Though many limitations exist in Wnt targeting fields, new trials are continuously ongoing. Moreover, clinical trials are underway for promising Wnt signaling-targeted drugs, such as DKK1 and FZD-targeting antibodies. Thus, hope still exists to develop the first Wnt-targeting drug. Furthermore, new advanced technologies are being developed to turn ‘undruggable’ proteins into ‘druggable’ proteins. Here, we introduce a new wave of Wnt-targeting strategies that makes use of the latest cutting-edge technologies.PROteolysis TArgeting Chimera (PROTAC) is a bifunctional molecule composed of an E3 ligase, a protein of interest (POI), and a linker capable of linking two ligands .The E3 ligase ubiquitinates POIs that interact with PROTAC, and the target protein is eventually degraded by the proteasome . PROTAC −/− organoids. Additionally, xStAx-VHL inhibits the growth of CRC xenografts and APC Min/+-driven intestinal tumors [In Wnt-targeted cancer therapy using PROTAC, β-catenin is an attractive intracellular target. xStAx-VHL is a PROTAC-targeting β-catenin consisting of xStAx and VHLL  Figure . xStAx il tumors .Since PROTAC does not work based on equilibrium occupancy, the working doses of PROTAC are very low, i.e., nanomolar concentrations . Thus, PA small molecule that induces or stabilizes the neo-interaction between two different proteins is called a ‘molecular glue.’ Some molecular glues, such as PROTAC, promote targets’ degradation by inducing new interactions between E3 ligase and target proteins. Additionally, molecular glue could complement PROTACs with far more advantages in terms of molecular weight than PROTACs . In thisRecently, various protein degradation technologies, such as PROTAB (proteolytic-targeting antibody), AUTOTAC (AUTOphagy-TArgeting Chimera), and LYTAC (lysosome-targeting chimeras), have been developed ,164. TheIndeed, a recent study showed a new targeting strategy, PROTAB, that uses Wnt signaling components. PROTAB is an antibody that induces the proteolysis of extracellular target receptors by linking to transmembrane E3 ligase ZNRF3, a negative regulator of Wnt signaling . A ZNRF3An antibody–drug conjugate (ADC) consists of a monoclonal antibody (mAB), a cytotoxic drug (payload or warhead), and a chemical linker . ADCs inSeptuximab vedotin (F7-ADC) is an ADC for targeting FZD7  Figure . It compPTK7, involving both Wnt and VEGF signaling, is associated with cancer drive and repression . In the LGR5-targeted ADCs, LGR5–MC-vc-PAB–MMAE and LGR5–NMS818, were recently developed. These LGR5-targeted ADCs target the LGR5-expressing population of tumor-initiating cells or cancer stem cells (CSCs)  Figure . These cCurrently, there are still difficulties in applying ADCs to cancer treatment. Most approved ADCs have side effects such as hematotoxicity . MoreoveOligonucleotides are emerging therapeutics that use small nucleotides (15–30 bp) with chemical modifications mainly to regulate the target gene’s transcription . These smiRNAs and siRNAs consist of short sequences to target RNA through partial base pairing . In prosAntisense oligonucleotides (ASOs) are single-stranded DNA analogs that consist of 16–22 bases . ASO binSeveral ASOs that target the Wnt pathway indirectly or directly are undergoing preclinical studies. LNA-modified ASO is reported as an indirect targeting ASO for the Wnt pathway . The lonβ-catenin is directly targeted by 2’-O-methoxyethyl chimeric ASO. Treatment with this β-catenin-targeted ASO showed a specific decrease in β-catenin expression in liver and white adipose tissue in high-fat-fed C57BL/6 mice  Figure . These dDespite the contribution of Wnt signaling to tumorigenesis being obvious and Wnt signaling inhibition having shown significant effects in preclinical models, no Wnt signaling-targeted drugs have proved clinically successful in cancer or other diseases ,6. The mIn addition to these efforts, combining the newest targeting strategies, such as PROTAC/molecular glue, ASOs, and ADCs, was recently introduced to Wnt-targeted therapy. Adopting these technologies and accumulating knowledge on mechanisms of action from past and current Wnt-targeting trials have allowed for a new wave of Wnt signaling-targeting therapies.These new targeting technologies could enhance the specificity of drugs. PROTACs eliminate their targets; they do not inhibit them. ADC combines the advantages of antibodies and cytotoxic drugs, ameliorating conventional side effects. The design of ASOs is straightforward and assigns specificity toward targets. These three methods can help improve the limitations of existing targeting strategies. Additionally, numerous combinations of these new techniques are possible.ASOs were first suggested in the early 2000s. However, the initial ASO system had limitations: poor cellular delivery, not being sufficiently stable under degradation by nuclease, and off-target effects. Additionally, some modified ASOs show high toxicity . OvercomIn conclusion, before the advent of third-generation ASOs and FDA-approved ASOs, it was assumed that ASOs could not be used for treatment. Twenty years ago, before the success of imatinib, the first FDA-approved kinase drug , people


Equine obesity is a significant health and welfare concern. The proportion of domestic horse populations that are overweight are as high as 45%. As the primary decision-makers for their horses’ care, owners are theoretically ideally placed to identify whether their horses are appropriately conditioned, however, research in other countries has shown that many owners are unable to accurately judge their horse’s body condition. In this study, through the comparison of body condition scoring (BCS) performed by an expert and the horse owners and interviews with owners, we aimed to identify the proportion of horses that were overweight or obese, to assess the accuracy of horse owners' BCS assessment both prior to and after receiving information and instructions on body condition scoring, and to identify common themes amongst owners’ views regarding BCS assessment and the Henneke BCS system.P < 0.001; ICC = .502, P < 0.001) and after  receiving information and instructions on scoring. Three quarters of the owners who took part in the study did not use any method of monitoring their horse’s body condition. Thematic analysis of owner responses was varied, with the most common theme being an awareness of the need to monitor or make changes to their horse’s condition with responses in this theme split between owners who felt in control and those who did not. Owner feedback on the utility and useability of the scorning system was that it was useful however parts are too technical or need improvement.Forty-five percent of the horses in this study were overweight or obese. The agreement between the owners and an equine veterinarian regarding the horses’ BCS was fair to good both prior to (κ = .311, Equine obesity is a significant problem in this population in Ireland. Horse owners’ ability to accurately judge their horse’s condition does not improve with provision of instructions on body condition scoring. These results combined with owners’ feedback on the Henneke BCS system indicate that it is not a tool that can be reliably used by owners.The online version contains supplementary material available at 10.1186/s13620-023-00237-w. Obesity is considered a significant health and welfare issue for horses , 2. DataAs the primary decision-maker for their horses, horse owners are theoretically ideally placed to identify health issues and seek veterinary advice early. With regards to body condition, identification of potential problems  in a timely manner, as necessary for adequate welfare , may be There are two equine body condition scoring systems commonly used; the 9-point Henneke system  and the In this study, we aimed to ascertain whether owners in Ireland can accurately judge their horses’ body condition, the effect on this ability of owners receiving instructions on scoring, and the proportions of horses who are underweight, appropriately conditioned, overweight and obese.This study was exempted from full ethical review by UCD’s Human Research Ethics  and Animal Research Ethics  Committees.A post was shared on the UCD Veterinary Hospital’s Facebook page asking for owners from counties Dublin, Kildare, Meath and Wicklow to sign up on a first-come first-served basis to a study on owners’ perceptions and decision making regarding their horse’s body condition, management, feeding, supplement use etc. Where an owner had more than one horse, they were asked to choose with which one they would like to take part.Upon arrival, the assessment process and what it would involve for both the owner and horse were explained to the owner and the owner was informed that they could cease participation at any point. Verbal consent was obtained prior to the questionnaire and clinical exams starting. One of the researchers interviewed the owner using a set questionnaire whilst the other conducted a basic examination of the horse, including assessing body condition using the Henneke body condition scoring scale. The latter researcher (referred to hereafter as the “expert”) assessing body condition was an experienced equine vet; this person was the expert for all horse assessments in this study. The exam and the expert scoring were conducted out of sight and hearing of the owner and of the researcher conducting the questionnaire portion of the visit.th January 2019 by University of Kentucky College of Agriculture, Food & Environment. https://thehorse.com/164978/body-condition-scoring-horses-step-by-step/ (accessed 24/06/2021). This article was used because it: was present in the first page of results when searched using the search term “body condition scoring horses” and therefore a page that would possibly be found by owners searching for information; included the scoring chart; was concise; and, explained the process and what to look for with regards to fat build-up in the six different body areas.During the visit, owners were provided with a two-page information sheet on the Henneke body condition scoring scale and an explanation of how to apply it to their horse . Intraclass correlations  were used to determine the absolute level of agreement between the expert and the owners. Cohen’s linear weighted kappa was used to assess the level of agreement taking the extent of differences between the expert and owner into account. Both tests were used for comparisons between the expert and owner scores both before the owners had received information and instructions on scoring and afterwards. Chi-square tests were used to assess association between BCS (Body Condition Score) and variables including management details, exercise routines and weight and condition monitoring. Owner responses given in addition to their answers to the open-ended survey questions were recorded in writing and entered into a master data sheet with the responses to the closed-ended questions. Responses to questions regarding monitoring and judgment of their horse’s body condition were analysed. Initial codes were identified after familiarization with the responses. An inductive approach , wherebyth June 2021 and Sunday 11th July 2021.The first 60 owners, or groups of owners where a single owner was acting on behalf of a group, who contacted the author in response to the Facebook post and whose horses were located in any of the four counties specified were included in the study. They were visited on dates between Monday 28The horses and ponies included in the study were of the following breeds/types: 10 cobs; nine Connemara ponies; seven Thoroughbreds; seven sport horses; four Irish Draught horses; one sport pony; one each of Selle Francais, Dutch Warmblood, English Warmblood, Holsteiner, New Forest Pony, Shetland and Welsh Cob; crosses including one Irish Draught cross, one Thoroughbred cross, one Irish Draught x Thoroughbred, one Holsteiner x ISH and one mixed native pony; and 10 with unknown, uncertain or multiple (more than two) breed heritage.Ages ranged from three years old to one pony in its 30s (exact age unknown). Heights ranged from eight and a half hands high to 18 hands high. There were 20 mares, 39 geldings and one stallion.Thirty-eight were kept on livery (including two DIY livery); 19 were kept privately; two were kept on livery yards but belonged to the yard owner or manager; and one was kept at a premises belonging to a friend of the owner. The number of horse-owner combinations surveyed within the same yard ranged from two to eight.The number of horses judged by the expert to be at each of the nine body condition levels can be seen in Fig. No significant associations between BCS and management details, exercise routine or weight and condition monitoring were found.Fifty-three (88.33%) horses were clinically normal on the day of examination. One was found to have enlarged sub-mandibular lymph nodes and six had increased digital pulse(s). Seven had a history of laminitis and eight had a history of colic; of those, one had a history of both. In addition, one animal was described as being pre-laminitic and one as “a bit colicky once”. Of those animals with a history of colic and/or laminitis, five had other health issues (confirmed and suspected). Of the horses and ponies with no history of laminitis and/or colic, 23 horses had other health or behavioural issues. The confirmed and suspected health issues reported, other than laminitis and colic, included ulcers, Pituitary Pars Intermedia Dysfunction, respiratory problems, lumps, musculoskeletal problems (including arthritis), skin conditions (including sarcoids), shivers and accidental injuries however these were not explored further in this study.Of the 60 horses and ponies included, 49 were reported as having up-to-date flu and tetanus vaccinations. With regards to de-worming, one was reported as being de-wormed once a year; 26 twice a year; six three times a year; 10 four times a year; and one five times a year. Of the remainder, 10 were de-wormed based on FEC results; one recently de-wormed twice in quick succession due to the horse looking “ribby” and one based on consistency of the droppings; three were unspecified; and one was unknown to the owner. Fifty-one owners reported their horse being seen by an Equine Dental Technician at least once a year and 46 reported their horse being seen previously at least once by at least one allied health practitioner  in their time owned by the owner.n = 43, 71.66%), 12 were provided with haylage, 29 with hay, and two with soaked hay. Forty-eight horses and ponies were given balancer and/or feed, and 37 received at least one feed supplement.Just under 50% of the horses and ponies included were living out at the time of their visits with the remainder stabled and turned out for a portion of the day or else stabled during the day and turned out overnight. Of the animals provided with forage  to hard  work.Six owners reported using a weight tape to monitor their horse’s weight whilst two owners reported using a specific body condition scoring system to monitor condition. One yard had a weighing scale and three of the eight owners surveyed at that yard reported using the scale to monitor their horse’s weight; four owners monitored condition by taking regular photos. Despite the majority (45) of owners reporting not using a specific method of monitoring weight or condition, of those, 37 said that they would note changes in their horse’s condition in specific areas Fig. .Fig. 2NuThe number of horses indicated by their owner on the numerical rating scale to be at each of the nine body condition levels, and the number of horses judged by them to be at each of the nine body condition levels after receiving information and instruction on the Henneke scoring system can be seen in Fig. P < 0.001)  mind.” Owner 14 expressed that “[horse] is so happy with field grass” while Owner 40 said they had considered using a grazing muzzle for weight but that it “seems cruel.”Two themes were identified within owners’ feedback regarding the body condition scoring system. The first of these was that the system was useful for assessing or monitoring body condition. Within this theme, owners expressed feedback that the scale, or parts of it, were useable or useful or that it included body areas they hadn’t considered assessing. There was also an element of the use of the system making the owners re-think their previous judgement; Owner 27 said that using the system made them look at their horse differently and Owner 14 that it made them “think twice” regarding their horse’s condition.The second theme identified was that the scale could be improved. Within this theme, some owners were critical of specific parts of the scale, highlighted that it in their opinion it wasn’t applicable to all breeds, types, ages or sexes or suggested specific ways of improving the useability of the scale including the addition of illustrations or photos. Other owners felt that the scoring system is too technical or difficult to use, either in whole or in part, with some owners who expressed the “too technical” opinion stating that they were unfamiliar with some body parts which were being referenced ; others expressed concern that there is no difference between descriptive terms such as “spongy” and “soft”, or that the different scores were too close together. Owner 60 stated that the scale needs to be “more idiot-proof.”The expert scores and initial owner judgements from the three yards with the highest numbers of participating horse-owner combinations were examined Table .Table 3CIn Yard A, three quarters (75%) of horses surveyed were in appropriate condition , with 37.5% being in optimal condition . Owner accuracy in Yard A was 50% . None of the owners in Yard A underestimated their horse’s condition however half overestimated by one point. Yard A participants’ horses consisted of mostly of high-level competition horses however two (one pony) were not competing and one of these was not being ridden at the time due to a diagnosis of gastric ulcers. Yard A had a weighing scale on site although not all owners reported using it, with some reporting using a weight tape or assessing visually or by how the horse feels when riding. All horses received different types and amounts of feed. The owners of horses 7 and 8 reported being aware of their horses’ need to lose weight. For those in work, exercise intensity ranged from 1 easy and 2–3 medium sessions per week to 1–2 easy, 4–5 medium and the occasional hard work session per week. Turnout for most horses was all day but was reduced for horses who required it for weight or other reasons. Participants in Yard A expressed comments within the “awareness and feels in control,” “weight balanced against other factors” and “awareness but doesn’t feel in control” themes, with the first of those themes being most prevalent among the owners in this group.In Yard B, all horses were in appropriate condition, although none were in optimal condition. Owner accuracy was 40%. None of the participants in Yard B underestimated their horse’s condition however 60% overestimated by one or two points. Yard B participants’ horses were used for dressage, showing and showjumping if competing, although to a lower level than those in Yard A. Two owners in Yard B reported taking photos to monitor condition. All horses received different types, or combinations of types, and amounts of feed. Exercise intensity ranged from 2 easy and 2 medium sessions to 1–2 easy and 4–5 medium sessions per week Turnout arrangements differed between horses, ranging from no turnout to living out full time in summer. Participants in Yard B expressed comments within the “awareness and feels in control” and “seeks expert opinion” themes, with the first of those themes being more prevalent among the owners in this group.In Yard C, two thirds of horses were over-conditioned, with one third being appropriately conditioned; none were in optimal condition. Owner accuracy in Yard C was 12.5%. Half of the owners in Yard C underestimated their horse’s condition by two or three points, with all believing their horses were in optimal condition, whilst a third underestimated their horse’s condition by one or two points. The horses and ponies in Yard C were used for pleasure riding with one being retired. None of Yard C’s owners reported using any specific means of monitoring their horses’ condition despite one mentioning that the yard owner uses a BCS scale. Most of the horses in Yard C were either fed a coarse mix or received no hard feed. Exercise intensity for the non-retired horses ranged from 1–2 easy and 1–2 medium sessions to 5 medium sessions per week. Two of the horses were stabled with the others living out. Participants in Yard C expressed comments within the “awareness but doesn’t feel in control,” “awareness and feels in control” and “seeks expert opinion” themes, with the first of those themes being most prevalent among the owners in this group.n = 27) of horses were overweight or obese. Owners’ ability to accurately judge their horse’s condition was fair, with owners of overweight horses underestimating their condition and owners of underweight horses overestimating their condition. Providing owners with a descriptive scoring scale and instructions as how to carry out body condition scoring had very little effect on the accuracy of their judgements. Owners’ actions and feelings regarding monitoring and managing their horses’ condition were varied. Only a quarter of owners used some method of regularly monitoring their horse’s condition. Owners’ feedback on the utility and useability of the body condition scoring scale were mixed with an overall trend towards the concept of the system and specific parts of it being useful but that parts were too technical.In this study, 45%  of monitoring condition, although the majority did state that they would notice weight gain or loss in specific areas. What is notable is that the body area mentioned most by owners in this study was the belly, which is not assessed in either of the two BCS systems commonly used; however belly girth measurement has been found to be a reliable indicator of weight and weight loss , 25. It Similarly to a recent study of horse owners in Italy , agreemeBody condition scoring may not be accurately useable by a significant proportion of horse owners, since agreement between the expert’s scores and the owners’ judgements was still classed as fair agreement despite the use of the scoring scale. A similar lack of change in accuracy has been seen in dog owners where the use of a BCS chart did not improve the accuracy of their judgement of their dogs’ condition, particularly that of overweight dogs . This faThat a proportion of owners are aware of issues or potential issues with their horses’ body condition and that this awareness is accompanied by a feeling of control is a positive finding. There remain owners who have similar awareness but do not feel in control. Some of these view condition as an aspect of the horse that they cannot influence, for example an older horse being underweight because of its age or a pony being overweight because of its breed. The potentially obesogenic environment in which an owner keeps their horse   and poteThe most challenging theme with regards to changing owner perceptions of overweight horses and ponies is likely that of not considering condition or weight to be important or as important as other factors. Previous research has identified that owners can perceive weight-loss measures, such as less food and/or more exercise, as negatives in terms of welfare .Our findings would suggest that Henneke system is not a reliably accessible body condition scoring tool for owners to use. The lack of significant improvement in the accuracy of owners’ judgement of their horses’ condition after receiving information on and instructions as to the use of the Henneke BCS system supports this conclusion. It is possible though, given the problems that appear to be common in the use of subjective scales such as BCS scales by horse and pet owners , 6, 26 tThe identification of potential yard cultures was a notable outcome of this study. Previous research , 14 has There were some limitations to this study. The actual BCS of each horse was determined by a single equine vet; it may have been better to have had multiple equine vets score each horse and use a consensus score. Certain criteria, such as exercise and amount fed, may have been under- or over-estimated by the owners in giving their answers. For owners with more than one horse, given the inclusion of the clinical exam, they may have been more likely to choose one with a history of poorer health or that they had particular concerns regarding. While the self-selecting nature of the participants ensured no bias from the researchers, it did result in far fewer underweight horses included than overweight horses. Finally, it is possible that training from the expert in how to perform the body condition scoring would have elicited better results in the post-instruction scoring however the researchers elected not to do this as it is not a feasible real-world solution.The results of this study, both qualitative and quantitative, strongly suggest that the use of body condition scoring by owners does not elicit accurate assessments of their horses’ condition. Further research, into alternative methods of equine body condition assessment by owners is warranted. Rendle et al. (2018) suggest that “[i]n the future, a combination of objective measurements may prove to be easier and more robust to apply than the BCS system for owners” however they state that more work would be required for validation, which is still the case. The high proportion of horses and ponies that were identified as overweight or obese, and the health consequences that brings, makes this a matter of some urgency. In the meantime, responsibility for identifying overweight horses, making sure owners are aware of the risks and enabling them in making changes, perhaps using the specific resources mentioned above, to facilitate weight loss remains with equine veterinarians, nurses and other professionals.Additional file 1.Additional file 2.


Hepatocellular carcinoma (HCC) is one of the deadliest cancers due to multifocal development and distant metastasis resulting from late diagnosis. Consequently, new approaches to HCC diagnosis and treatment are required to reduce mortality rates. A large body of evidence suggests that non-coding RNAs (ncRNAs) are important in cancer initiation and progression. Cancer cells release many of these ncRNAs into the blood or urine, enabling their use as a diagnostic tool. Circular RNAs (CircRNAs) are as a members of the ncRNAs that regulate cancer cell expansion, migration, metastasis, and chemoresistance through different mechanisms such as the Wnt/β-catenin Signaling pathway. The Wnt/β-catenin pathway plays prominent roles in several biological processes including organogenesis, stem cell regeneration, and cell survival. Aberrant signaling of both pathways mentioned above could affect the progression and metastasis of many cancers, including HCC. Based on several studies investigated in the current review, circRNAs have an effect on HCC formation and progression by sponging miRNAs and RNA-binding proteins (RBPs) and regulating the Wnt/β-catenin signaling pathway. Therefore, circRNAs/miRNAs or RBPs/Wnt/β-catenin signaling pathway could be considered promising prognostic and therapeutic targets in HCC. This trNon-coding RNAs (ncRNAs) are RNAs derived from the human genome but not translated into protein . NcRNAs The Wnt family includes a group of proteins that play important roles in a various cellular functions, such as organ development, stem cell renewal, and the cell viability . The WntSeveral studies have found that circRNAs have the potential to be used as biomarkers in tissue samples for cancers diagnosis, with noticeable clinical applications for predicting HCC prognosis. This could potentially lead to improved therapeutic strategies , 26. ForOur theory is that circRNAs could become a novel approach for detecting and treating HCC in the future. To support this idea, we conducted a review of recent studies that investigate the impact of circRNAs on HCC development, with a particular focus on how they affect miRNA regulation and the Wnt/β-catenin signaling pathway.2. Many studies have investigated the role of Wnt signaling in the tumorigenesis, with an focus on signaling that relies on β-catenin , and the calcineurin phosphatase. In addition to Ca2+-dependent Wnt STP, tumor cells have been found to have a novel non-canonical Wnt STP mediated by FYN tyrosine kinase and STAT3 : canonical (Wnt/β-catenin) and non-canonical (planar cell pole (PCP) and Wnt/calcium (Wnt/Ca2+)) , 30 as s-catenin . In the -catenin . β-caten-catenin . Recepto-catenin . Therefo-catenin . This caolipases . When thnd STAT3 . Wnt prond STAT3 .3A (the most abundant type), intronic circular RNA, Exon-intron circular RNA, Intragenic circular RNAs, and Antisense circular RNAs , Lens culinaris-agglutinin-reactive fraction of AFP (AFP-L3), protein induced by vitamin K absence orantagonist-II (PIVKA-II), vascular endothelial growth factor (VEGF), hypoxia-inducible factor (HIF), and others . On the 5CircRNAs have been demonstrated to modulate cell proliferation, migration, and apoptosis by regulating gene transcription or by activating or inactivating signaling pathways associated with cancer . The mos5.1in vitro and in vivo  has been shown to be overexpressed in breast cancer and glioma , 88. Liu5.5The expression of hsa_circ_0004018 was decreased in HCC compared with non-tumor tissue . Accordi5.6in vitro (Previous studies have indicated that circRNA zinc finger protein 292 (circZNF292) was upregulated in the hypoxic environment in the solid tumors, including HCC. CircZNF292 has proangiogenic function in the hypoxic condition of tumor environment in vitro –93. It hin vitro . It has in vitro , 96. Cirin vitro . Thus, cin vitro .5.7It has been revealed that hsa_circRNA_104348 expression increased in the HCC and correlated with the poor prognosis. Overexpression of hsa_circRNA_104348 induced proliferation, whereas inhibiting apoptosis of the HCC cells . Rhoteki5.8Sorafenib is a kinase inhibitor drug which is used in the advanced HCC . CircRNA5.9Circular RNA circZFR has been linked to the progression of HCC, breast cancer, papillary thyroid carcinoma, and bladder cancer –105. Cir5.10in vivo and in vitro could repress the progression, migration, and invasion of HCC cells. Interestingly, circβ-catenin is a translatable protein that has resulted in an isoform known as β-catenin-370aa. This isoform could attenuate glycogen synthase kinase 3β (GSK3β) function through prevention of GSK3β binding to fill-length β-catenin. GSK3β inhibits β-catenin activity by β-catenin degradation. Thus, circβ-catenin could enhance β-catenin level in the tumor microenvironment by preventing β-catenin destruction, inducing Wnt/β-catenin signaling pathway (CircRNA-0004194, also known as circβ-catenin, was overexpressed in the HCC. Liang et al., observed that circβ-catenin silencing  pathway , 107.5.11The studies have shown that the expression of circZKSCAN1 was decreased in the HCC cell lines in comparison to the non-tumor tissue . Previou5.12in vivo model, intratumoral injection of a miR-541-5p inhibitor reduced tumor size. As a result, the circMTO1/miR-541-5p/ZIC1 axis may have therapeutic potential for HCC by suppressing HCC cell growth, invasion, and metastasis through regulation of the Wnt/β-catenin signaling pathway (Previous studies were stated that circMTO1 play a role as a tumor suppressor  and inhi pathway . CircMTO pathway . Downreg pathway , 118 and pathway .6Clinicians have believed that treatment choices require early prognostic data. A growing body of evidence suggests that Wnt-related circRNAs could be valuable for prognosis . In seve7NcRNAs are play a promising role in regulating different types of cancers includes HCC through the regulation of gene expression and modulation of biological pathways. In this context, targeting ncRNAs, include circRNAs, represents a novel method in cancer treatment and control drug resistance with advantages compared to conventional therapeutic strategies, such as specificity and potential reduction in toxicity , 127. FoThe Wnt signaling pathway is required for several physiological processes, including cell differentiation, motility, expansion, and muscular tissue formation. Both circRNA and the Wnt signaling pathway play important roles in cancer initiation and development. Developing evidence suggests that the circRNA/Wnt axis regulates tumor advancement, invasion, and metastasis by modulating the expression of cancer-associated genes. CircRNAs that are involved in the Wnt pathway typically perform their function by sponging miRNAs, which can be used as diagnostic markers. Wnt-related circRNAs may potentially be useful as biomarkers in the therapy of HCC. To manage tumor growth, researchers are attempting to either upregulate tumor-promoting circRNAs or downregulate tumor-suppressor circRNAs. Further research is needed to confirm the interactions and related mechanisms between suppressive miRNAs and corresponding circRNAs involved in the Wnt pathway for the diagnosis, prognosis, and treatment of HCC.The authors confirm contributions to this paper as follows,AM, conceptualization, investigation, database searching, writing, and review manuscript. FZ, ST, HR, MM, YG, NH, SH, SG, RM, BG, and ZA, data collection, writing, draft preparation, and figure design. MA and HM contributed in the project administration, editing, validation, coordinated the study, and accurately evaluated the manuscript. All authors contributed to the article and approved the submitted version.


Dysregulated expression of specific non-coding RNAs (ncRNAs) has been strongly linked to tumorigenesis, cancer progression, and therapeutic resistance. These ncRNAs can act as either oncogenes or tumor suppressors, thereby serving as valuable diagnostic and prognostic markers. Numerous studies have implicated the participation of ncRNAs in the regulation of diverse signaling pathways, including the pivotal Wnt/β-catenin signaling pathway that is widely acknowledged for its pivotal role in embryogenesis, cellular proliferation, and tumor biology control. Recent emerging evidence has shed light on the capacity of ncRNAs to interact with key components of the Wnt/β-catenin signaling pathway, thereby modulating the expression of Wnt target genes in cancer cells. Notably, the activity of this pathway can reciprocally influence the expression levels of ncRNAs. However, comprehensive analysis investigating the specific ncRNAs associated with the Wnt/β-catenin signaling pathway and their intricate interactions in cancer remains elusive. Based on these noteworthy findings, this review aims to unravel the intricate associations between ncRNAs and the Wnt/β-catenin signaling pathway during cancer initiation, progression, and their potential implications for therapeutic interventions. Additionally, we provide a comprehensive overview of the characteristics of ncRNAs and the Wnt/β-catenin signaling pathway, accompanied by a thorough discussion of their functional roles in tumor biology. Targeting ncRNAs and molecules associated with the Wnt/β-catenin signaling pathway may emerge as a promising and effective therapeutic strategy in future cancer treatments. Cancer is a significant global public health issue, with a continuously increasing incidence and mortality rate. It imposes a tremendous burden on both individuals and societies worldwide . The devRecent studies have shown that non-coding RNAs (ncRNAs), as well as Wnt/β-catenin signal pathways, play an active role in the development and progression of cancer . ncRNAs This review aims to summarize the impact of abnormal expression of ncRNAs related to the Wnt/β-catenin signaling pathway on cancer initiation and development. It intends to elucidate the regulatory relationship between ncRNAs and the Wnt/β-catenin pathway. Additionally, it aims to provide an overview of the current application value and potential of ncRNAs in cancer diagnosis and treatment, thereby offering further insights for future research on the role of ncRNAs in tumor initiation and therapeutics. The ncRNAs data were retrieved through a comprehensive search on NCBI (PubMed), employing specific keywords, such as “ncRNA”, “Wnt/β-catenin”, and various types of cancer like “breast”, “lung”, “colorectal”, “prostate”, and “gastric cancer.” The search results were further filtered to encompass publications between the years 2019 and 2023.ncRNAs, a special class of RNA transcripts, do not encode proteins but exhibit high specificity and stability, making them crucial for assessing the occurrence and progression of cancer ,24. BaseBoth lncRNAs and circRNAs are over 200 nt in length; can be transcribed from the exon, the intron, the intergenic region of the gene, and the 5/3-untranslated region; and fold into complex secondary structures that facilitate their interactions with DNA, RNA, and proteins ,33,34,35The Wnt signaling pathway is an important pathway widely present in multicellular organisms. It plays a crucial role in biological development, tissue regeneration, and various diseases, particularly in the occurrence and progression of cancer ,42,43. F2+ signaling pathway, and the planar cell polarity (PCP) pathway [Upon activation of the Wnt signaling pathway, downstream signaling requires the further activation of DSH. DSH consists of three distinct structural domains: the amino-terminal DIX domain, the central PDZ domain, and the carboxy-terminal DEP domain. These three domains of DSH play a crucial switch-like role in the Wnt signaling pathway  and dete pathway ,46. EachCurrently, research on the Wnt pathway primarily focuses on the Wnt/β-catenin signaling pathway branch. This pathway can determine cell fate, cell proliferation, cell survival, and intercellular interactions . DysreguThe interplay between ncRNAs in regulating the Wnt/β-catenin signaling pathway is depicted in An increasing body of research has demonstrated the significant regulatory roles of ncRNAs in the occurrence and progression of various cancers through the modulation of the Wnt signaling pathway. The Wnt pathway is known to play crucial roles in cell differentiation, proliferation, and self-renewal in a variety of cells. Disturbances in the Wnt pathway have been closely associated with the development and progression of several cancers, including liver cancer, colon cancer, breast cancer, and prostate cancer. As a class of RNA molecules that do not encode proteins, ncRNAs have garnered extensive attention regarding their mechanisms and regulatory modes in cancer. The specific mechanisms of ncRNAs vary among different types of cancer. For example, in bladder urothelial carcinoma, miRNA-139-3p inhibits malignant progression by targeting KIF18B and inactivating the Wnt/β-catenin pathway . In contDespite the need for further research to gain a deeper understanding of the mechanisms by which ncRNAs contribute to cancer, the study of ncRNAs and their regulation of the Wnt pathway hold great significance for cancer diagnosis, treatment, and the development of personalized medicine in the future.In the past few years, ceRNA has been detected in Wnt/β-Catenin signaling-pathway-related cancers, exhibiting increasingly important regulatory roles , particuBreast cancer (BC), as a common disease threatening women’s health, has been receiving much attention in terms of its treatment and prognosis . The WntAs a small RNA molecule, miRNA can regulate gene expression at the post-transcriptional level by interacting with mRNA targets. It was found that miR-296-3p downregulated SOX4 by targeting the Wnt/β-catenin signaling pathway to produce anti-tumor effects in TNBC , while mIn addition, ncRNAs also play an important role in the treatment and resistance of BC. Apatinib can inhibit BC development by blocking the Wnt/β-catenin signaling pathway through downregulation of lncRNA ROR . In BC cUltimately, the ncRNAs associated with the Wnt/β-catenin signaling pathway plays an important regulatory role in BC. These ncRNAs affect the proliferation, invasion, metastasis, and stem-cell properties of BC cells by directly or indirectly regulating Wnt/β-catenin signaling. Moreover, they are involved in the regulation of drug resistance and immune escape in BC cells. A deeper understanding of the mechanisms of these ncRNAs in BC will help to reveal the molecular basis of BC development and provide new strategies and targets for precision therapy and personalized medicine. However, many questions still need to be further investigated, including the interaction of ncRNAs with the Wnt/β-catenin signaling pathway, its regulatory effects in the tumor microenvironment, and its potential and limitations in clinical application. Through continuous intensive research, we hope to reveal more details about the occurrence and development of BC and provide more precise methods and strategies for its treatment and prevention. The study of Wnt/β-catenin signaling pathway in lung cancer (LC) is increasingly valued, and the role of the ncRNA involved in this tumor is gradually revealed. Lung cancer is a highly heterogeneous and lethal disease, and understanding the function and mechanism of the Wnt/β-catenin signaling-pathway-related ncRNAs in lung cancer is essential to revealing the molecular basis of its occurrence and development and for the development of new therapeutic strategies . MiR-590 is a type of miRNA that commonly exhibits abnormal expression in various tumors. Ma et al. found that this miRNA is downregulated in non-small-cell LC (NSCLC) and closely related to patient prognosis . OverexpAnother ncRNA related to the Wnt/β-catenin signaling pathway is lncRNA SNHG11 . Its expSeveral other ncRNAs have also been found to be intimately involved in the initiation and progression with LC ,95,96,97Thus, Wnt/β-catenin signaling activation favors lung cancer progression. Potential therapeutic strategies could be focused on targeting Wnt signaling or ncRNA as an upstream mediator. The limitations of the current experiments are their emphasis on oncogenic ncRNA and the increased effort required to identify the tumor-suppressor ncRNAs regulating Wnt signaling in lung cancer.Colorectal cancer (CRC) is a common cancer with a high incidence in both men and women . DespiteMiRNAs are a class of short-chain RNAs with a length of 20–24 nucleotides that have been found to be important regulators in various tumors. Studies have found that miRNA-621 directly targets LEF1 and inhibits Wnt/β-catenin signaling, thereby playing a role in the inhibition of metastasis in CRC . Other tOverexpression of SNHG15, located on chromosome 7p13, has been associated with low survival rates in many human malignancies, including CRC ,106. RecCancer pathogenesis is correlated with lncRNA expression, and several approaches have been proposed to target lncRNA for cancer therapy . One sucProstate cancer (PCa) is a prevalent malignant tumor and remains one of the primary threats to male health . AlthougAmong these ncRNAs, certain microRNAs exert inhibitory effects on the development of PCa through mechanisms such as targeting SLC39A7 and suppressing the Wnt/β-catenin signaling pathway. For example, miR-15a-3p inhibits the proliferation, invasion, and EMT of PCa cells by targeting SLC39A7 and suppressing the Wnt/β-catenin signaling pathway, providing novel therapeutic targets for PCa treatment . MoreoveLncRNAs are another important category of ncRNAs in PCa. LncRNA SNHG7 is highly expressed in PCa cells and acts as a sponge for miRNA-324-3p. Through sponge effects, upregulated SNHG7 positively regulates WNT2B, while downregulation of SNHG7 inhibits EMT in PCa, further suppressing cell proliferation, invasion, and metastasis ; lncRNA In recent studies, there has been a growing interest among researchers in exploring the issue of drug resistance in PCa; there is also research focused on the issue of drug resistance in PCa. Currently, cisplatin is widely used in the treatment of PCa, but cisplatin resistance remains a significant obstacle to cisplatin-based chemotherapy. Studies have found that upregulation of miR-425-5p sensitizes human PCa to cisplatin by targeting GSK3β and inactivating the Wnt/β-catenin signaling pathway, providing a new target for treating PCa, particularly cisplatin-resistant PCa .Immune escape is an important feature in tumor development, which refers to the certain degree of escape ability of tumor cells to attack the immune system, which causes the tumor to be unable to be effectively cleared by the immune system. To suppress PCa progression, anti-tumor immunity is activated, for which cytotoxic T cells are essential. However, lncRNA can induce PD-1 expression, prevent cytotoxic T-cell proliferation and mediate their apoptosis, leading to immune escape from PCa . TherefoGastric cancer (GC) is a prevalent malignancy, one of the most frequent cancers worldwide, and its incidence and fatality rates are still increasing ,131,132.Research has revealed upregulated expression of certain miRNAs, such as miR-150, miR-876-5p, miR-15a-3p, and miR-216a-3p ,136, whiAdditionally, ncRNAs are involved in regulating the Wnt/β-catenin pathway, which has a vital role to play in the development and evolution of GC. For example, circCNIH4 and lncRNA NNT-AS1 inhibited apoptosis in GC cells by inactivating the Wnt/β-catenin pathway, while circSmad4, circHIPK3, circAXIN1, and LINC0035 promoted GC advancement and suggested poor prognosis through activation of the Wnt/β-catenin pathway ,145,146.Although numerous studies have evaluated the underlying molecular mechanisms associated with Wnt/β-catenin-mediated GC and investigated a number of treatments that inhibit this oncogenic pathway, elimination of several drawbacks in therapies targeting Wnt/β-catenin, such as drug resistance or non-responsiveness to signaling requires further investigation. As described in this study, many ncRNAs, as dual-acting regulators, can directly or indirectly activate or inhibit the Wnt/β-catenin pathway, and the interaction between these ncRNA and Wnt/β-catenin signaling pathways seems to be a key mechanism in regulating GC. Thus, targeting the circRNA/lncRNA/miRNA/β-catenin axis may be a promising alternative to other targeted therapies based on Wnt/β-catenin signaling.Despite significant progress in understanding the mechanisms of action of ncRNAs for Wnt/β-catenin signaling-pathway-associated cancers, many therapeutic challenges remain ,153. TheThe Wnt/β-catenin signaling pathway plays a critical role in various types of cancers, including colorectal cancer, breast cancer, and lung cancer. ncRNAs have been extensively studied and shown to play a key role in regulating cellular biological processes. Here are some suggestions for ncRNA-based therapies targeting the Wnt/β-catenin signaling pathway in cancer:miRNA-based therapy: Introducing miRNAs that inhibit the activation of the Wnt/β-catenin signaling pathway can effectively suppress the growth and spread of related tumors. For example, miR-34a and miR-200 families have been shown to inhibit β-catenin expression, thereby suppressing the Wnt/β-catenin signaling pathway. Currently, in phase I clinical trials, researchers have successfully delivered synthetic miR-34a mimics to the liver using lipid nanoparticle delivery systems, precisely targeting cancer tissues and regulating the expression of various tumor-suppressor genes such as p53. Therefore, synthesizing miRNA mimics or utilizing delivery systems carrying these miRNAs could be a potential therapeutic strategy.in vitro transfection or in vivo delivery of RNAi molecules. The CRISPR-Cas9 editing system can induce DNA double-strand breaks on the circular structure of circRNAs using specific oligonucleotide sequences, leading to their degradation. Additionally, the CRISPR-Cas9 system can be used to genetically edit key regulatory genes to further modulate the activity of the Wnt/β-catenin signaling pathway.circRNA regulation: circRNAs have also been found to be associated with the Wnt/β-catenin signaling pathway in cancer. Through techniques such as bioinformatics analysis and high-throughput sequencing, key circRNAs relevant to the Wnt/β-catenin signaling pathway can be identified and validated. Their expression levels and functions in cancer can be experimentally validated. In the functional studies and mechanism elucidation phase, researchers can determine the specific functions and molecular mechanisms of these relevant circRNAs in tumor growth, proliferation, metastasis, and drug resistance. Once the functions and mechanisms of key circRNAs are confirmed, therapeutic strategies targeting circRNAs can be considered. RNA interference (RNAi) and the CRISPR-Cas9 editing system are commonly used techniques for circRNA targeting. RNAi can guide the degradation or inhibition of circRNAs by designing and synthesizing specific small RNA molecules, such as siRNAs or shRNAs. This can be achieved through lncRNA-targeted therapy: Many lncRNAs have been found to play important roles in controlling the Wnt/β-catenin signaling pathway. By designing appropriate off-target approaches, the functions of these lncRNAs can be selectively interfered with to regulate the activity of the Wnt/β-catenin signaling pathway. Specifically, targeted interventions using RNAi technology and the CRISPR-Cas9 editing system can selectively reduce or inhibit the expression of relevant lncRNAs. For example, an engineered nanoparticle platform delivering lncAFAP1-AS1 siRNA effectively reversed the radioresistance of TNBC. In addition to nanoparticle-based drug delivery systems, recent research has also proposed the therapeutic potential of using the CRISPR/Cas9 system to edit LncRNA expression in CRC patients, and relevant exploration is underway.Combination therapy: Combination therapy is a strategy that integrates ncRNA-based therapy with other conventional treatment modalities, including chemotherapy, targeted therapy, and immunotherapy, aiming to enhance the effectiveness of cancer treatment. By simultaneously targeting the Wnt/β-catenin signaling pathway and other oncogenic signaling pathways, combination therapy can comprehensively target cancer and offer several advantages. Firstly, combination therapy can overcome resistance issues. Some tumors develop resistance to chemotherapy drugs, but by utilizing ncRNA-based therapy, tumor sensitivity to drugs can be enhanced, effectively addressing drug resistance. Secondly, ncRNA-based therapy and other conventional treatment modalities typically inhibit tumor growth and spread through different mechanisms. Therefore, combining them can achieve synergistic effects on multiple targets, leading to more effective control of tumor development. Additionally, ncRNA-based therapy generally has lower toxicity and side effects, so combining it with conventional treatment methods can reduce the side effects associated with therapy and improve the quality of life for patients. Finally, combination therapy employs a comprehensive treatment approach, combining different treatment modalities that can act on tumors at different levels and time points. This integrated treatment strategy can comprehensively suppress tumors and improve the overall effectiveness of treatment.The diverse mechanisms by which ncRNAs regulate the Wnt/β-catenin signaling pathway represent potential therapeutic targets for cancer treatment. Further research is needed to identify specific ncRNAs and their potential mechanisms in different types of cancer, as well as to develop more effective delivery strategies. Overall, studying ncRNAs in Wnt/β-catenin signaling-pathway-associated cancers has great potential to improve cancer diagnosis and treatment.