SoftMatter2016, 12, 658), suggesta non-monotonic temperature response. These theories propose thatthe “free” (dissolved) polymers will mediate attractivedepletion interactions at room temperature, with a concomitant clustering/flocculationat a sufficiently high polymer concentration. At high temperatures,where the solvent is poorer, this is predicted to be replaced by attractivebridging interactions, again resulting in particle condensation. Interestinglyenough, our theoretical framework, based on classical density functionaltheory, predicts an intermediate temperature regime where the polymer-mediatedinteractions are repulsive! This obviously impliesa homogeneous dispersion in this regime. These qualitative predictionshave been experimentally tested and confirmed in this work, whereflocs of particles start to form at room temperature for a high enoughpolymer dosage. At temperatures near 45 °C, the flocs redisperse,and we obtain a homogeneous sample. However, samples at about 75 °Cwill again display clusters and eventually phase separation. Usingresults from these studies, we have been able to fine-tune parametersof our coarse-grained theoretical model, resulting in predictionsof temperature-dependent stability that display semiquantitative accuracy.A crucial aspect is that under “intermediate” conditions,where the polymers neither adsorb nor desorb at the particle surfaces,the polymer-mediated equilibrium interaction is repulsive.In this work, wehave synthesized polystyrene particles that carryshort end-grafted polyethylene glycol (PEG) chains. We then addeddissolved 100 kDa PEG polymers and monitored potential flocculationby confocal microscopy. Qualitative predictions, based on previoustheoretical developments in this field (Xie, F.; et al.  Thismay facilitate a controlled stability.15 Crystallization of colloidal particles has also been investigatedin many soft matter studies, and the coexistence between “gas”and ”liquid” phases can often be regulated by polymerproperties.17 Previous work19 has established thermoresponsive aggregation, or cluster formation,in aqueous dispersions containing polystyrene (PS) particles, carryinga layer of grafted short polyethylene glycol (PEG) chains. A temperatureincrease from an ambient value, at which the dispersion remained stericallystabilized, caused a reduced solubility of the polymer grafts, whicheventually resulted in gelation, aggregation, or cluster formation.An interesting result in the work by Shay et al.19 was that gelation occurred at temperatures far below thelower critical solution temperature (LCST) of an aqueous PEG solution.Colloidal dispersionshave been the subject of many studies, withfocus ranging from stability and drug delivery, to self-assembly,patch formation, and so on.24 In this case, the origin of the net attraction is an osmotic pressuredifference as polymers are depleted from the configurationally restrictedinterparticle regions. Adsorbing polymers can also mediate attractiveinteractions, but in these cases, the underlying mechanism is insteadattractive polymer bridges connecting the particles. The length scaleof forces generated by polymer addition can be varied by the degreeof polymerization, the concentration, and the polymer structure.17The addition of non-adsorbing polymers to particle dispersionscan also generate aggregation.22 reported interesting re-entrant flocculationbehavior of colloidalparticles in an aqueous dispersion containing polymers. Several differentpolymer and particle combinations were investigated, but the onesdisplaying the re-entrant behavior had one common property: a solvent(water) that becomes poorer (for the polymer) as the temperature wasincreased. In those systems, a crystalline colloidal phase at lowtemperatures melts to a homogeneous fluid upon increasing the temperature.A further temperature increase, however, eventually gives rise toa flocculated phase. We have recently developed a model for such systems.25 In this model, monomers are assumed to be ineither of two classes of states , labeled A and B, where B is more solvophobicthan A. On the other hand, the degeneracy of the B class exceeds that of A; that is, the populationof solvophobic monomers increases with temperature. This can leadto an LCST.28 Adding solvophobic colloidal particles to such asolution generates a system that displays the same qualitative temperatureresponse as was observed by Feng et al. That is, at low temperatures,type A monomers predominate and one observes depletioninteractions, whereas polymer bridging dominates at higher temperaturesdue to the attraction between type B monomers and thecolloidal surface. Interestingly, the intermediate temperature regimeis characterized by a polymer-mediated potential of mean force (PMF)between colloidal particles, which is repulsive.This is a crucial and general theoretical observation, which was verifiedfor systems for which one can obtain exact solutions. As surfaces are gradually modified from being nonadsorbingto adsorbing, there is an intermediate regime where polymer-mediatedinteractions indeed are repulsive. In other words, interactions betweencolloidal particles, mediated by neutral nongrafted polymers, generallyfollow the trend: attraction → repulsion → attractionas the surface affinity toward monomers proceeds from repulsive toattractive.29In a recent publication, Feng et al.29 we here aimfor a sort of theory–experiment loop. That is, armed with thetheoretical progress described above, we can make qualitative predictionsfrom which we design an experimental system. The measured temperatureresponse of the latter will then permit the construction of a moretargeted (semiquantitative) theoretical model by the adjustment ofa few parameters. Specifically, we will perform measurements on PSparticles carrying short (2 kDa) PEG chains grafted onto their surfaces.These are dispersed in an aqueous dispersion that also contains dissolved(“free”) PEG polymers, having a molecular weight of100 kDa. The A/B state description of PEGmonomer rotational isomers does seem to capture the essential physicsunderlying the presence of an LCST in aqueous solution. However, whilethe model is elegant, it is also somewhat complicated, and it mightbe desirable to simplify the model to one with fewer parameters, atthe expense of a less detailed treatment of separate entropy/energycontributions. In this work, we have chosen to describe the effectivedegree of hydrophobicity of the PEG monomers, via the strength ofa simple Lennard-Jones (L-J) potential, acting between all monomers.The L-J potential acting between the monomers will then be regardedas an interaction free energy (PMF) in this model.While we have not made attempts to establish quantitative relationsbetween this L-J potential and a specific A/B model, the qualitative predictions by the latter do suggest moreattractive L-J interactions at elevated temperatures even comparedto thermal energy (kT). Using the simpler L-J description,we do sacrifice the mechanistic origin of the increased attraction(between monomers as well as between a monomer and a hydrophobic surface)at elevated temperatures.Inspired by our recentstudies,Guided by the general theoreticalpredictions described above,we anticipate depletion-generated flocculation at room temperature,where water is a good solvent, at sufficiently high concentrationsof dissolved (free) PEG. On the other hand, at high temperatures,PEG monomers become hydrophobic and thus attract. There is also anexpected affinity between these monomers and the hydrophobic barePS surface. Hence, bridging attraction should lead to flocculationat high temperatures. Finally, our theoretical considerations suggestthat there is an intermediate temperature regime where dissolved PEGchains mediate repulsive interactions, which should facilitate a stablehomogeneous dispersion.Confocal laser scanning microscopy (CLSM)and dynamic light scattering(DLS) will be used to visualize the structural properties of our dispersions.We believe that our suggested combination of experiments and modelingwill guide us toward a better understanding of interactions and temperaturedependence in polymer/particle dispersions.18 Using a dispersionpolymerization method, styrene monomers in the presence of an initiatorand PEG, with a molecular weight of 2 kDa, propagate to form PS particles,which then become surface grafted with short PEG. The presence of2 kDa PEG provides stabilization to the dispersion at room temperaturethrough steric repulsive forces between the grafted layers. In otherwords, the grafted PEG chains act as a stabilizing agent. Subsequentdialysis was performed to purify the dispersions.PS core–shell particles,grafted with PEG, were prepared according to the protocol describedin our previous work.PEG, withan average molecular weight of 100 kDa, was supplied by Sigma-Aldrich.Deionized Milli-Q water was used for all of the synthesis protocols.I, as a function of the wave vector, q, as well asthe lag time, τ, can be measured. A useful dynamic quantityis the autocorrelation function, g2, definedasq is given byn is the refractive index of the solvent. For a monodispersesystem, the function g1 can be definedas31c is the characteristicrelaxation time and γ is the Kohlrausch exponent. Note that D is the Stokes–Einsteindiffusion constant. Thehydrodynamic radius, Rh, can be then calculatedasDLS measurements were madeusing a Malvern Zetasizer equipped with a HeNe laser, with a wavelengthof λ = 633 nm, and a detector positioned at a scattering angleof 173°. A source of light shines through a dilute sample ofparticles. The intensity and fluctuation of the scattered light aremeasured and used to study the dynamic properties of the colloidaldispersions. The intensity of the scattered light, CLSM was used forimaging and analysis of cluster formations in the samples. CLSM snapshotswere monitored on a Leica TCS SP5 scanning microscope, operated inthe inverted mode, and equipped with a HeNe laser, λ = 543 nm.Using two cover glasses, separated by a 120 μm spacer, and a100×/1.4 NA oil immersion objective, imaging of samples was performedby operating the microscope in a bright-field mode.2.We recall that the experimental system is composed of PS particlescarrying grafted 45-mers (2 kDa PEG) and dissolved 2274-mers (100kDa PEG). In our earlier work, where these PEG-grafted PS particleswere synthesized, the grafting density, γ, was measured as γ= 0.355/nm32 The polymer-mediatedinteractions are short-ranged compared to the particle size, so theDA is expected to be nearly exact for these systems. These conditionsalso ensure that many-body interactions are negligible; that is, thesystem can be accurately described by pairwise additive PMFs actingbetween the colloidal particles. From CLSM images, we estimate anaverage particle diameter of 2300 Å.We shall adopt a flat surface geometryto work out free energiesper unit area and then the Derjaguin approximation (DA) to establisha PMF between the particles.33 utilized a hard-sphere descriptionof the monomers. The optimized diameter was found to be 2.65 Å.Neighboring monomers in a chain were connected by rotationally flexiblebonds, but with a fixed bond length of b = 4 Å.Since we are aiming for a temperature-dependent model, we have useda hard-sphere plus attractive L-J description of the monomers; thatis, all monomers interact via ϕ(r), definedasr isthe separation between the monomers. Guided by the low-temperature(good solvent) simulation work, we have set σ = 2.65 Å.Our PMF calculations are based on classical statistical–mechanical density functional theory (DFT); that is, they do not involve quantummechanical calculations. We have utilized the so-called generalizedFlory dimer approximation of excluded volume effects.35 The full polymer DFT formulation is quite lengthy but rather standardby now. We provide a condensed description below. DFT is usually formulatedsuch that nonlocal excluded volume effects are (approximately) accountedfor at a monomeric length scale. However, for polymeric molecules,intrachain correlations are not appropriately managed. The influenceof other monomers via excluded volume and soft interactions is treatedin a “smeared-out” manner, which in our flat surfacegeometry implies an assumption that the densities only vary with thedistance (z) normal to the surfaces but is uniformalong the lateral directions (x and y). This tends to be an accurate approach at high monomer densities,such as within the grafted layers. For instance, an earlier work demonstratedexcellent agreement between simulated (b* ≡ b/σ ≈ 1.5) and calculated monomer density profiles29 for fully flexible polymers, provided that theDFT calculations utilized a somewhat increased bond length, b* = 2. This is the bond length we have adopted for allpolymers in this work. However, in a dilute solution of isolated chains,the mean-field error is more significant. This is because the monomerdensity is locally high within “polymer occupied” regionsand vanishingly small between these regions. This means that a mean-fieldapproximation of a uniform monomer concentration can be rather poor.As a consequence, the predicted polymer “size” (Rg) of our “DFT polymers”is similar to that of ideal chains, which is more compact than theyshould be. This is because the local monomer density in reality tendsto be higher than the corresponding mean-field (“smeared-out”)average. An unfortunate, and well-known, consequence is that the predictedrange of polymer-mediated interactions is underestimated. One wayto correct for this is to increase the bond length. We have, however,chosen to retain the bond length and instead introduce a bond anglepotential between next-nearest neighbors in the ungrafted (dissolved)chains. We thus introduce a potential, EB, defined assi denotes the bondvector between monomers i and i +1 , and α is the strength of the bending potential.Thus, as α increases, the polymers will tend to “stretchout”. The value of α was adjusted so that the low densitymean-field prediction matches the one obtained from simulations inthe good solvent limit for our target 2274-mers. Specifically, wesimulated an isolated 2274-mer using the previously validated modelin the good solvent limit33  and calculated the average degreeof gyration, Rg = ⟨Rg2⟩1/2. Then we simulated an isolated 2274-mer with point-like monomers that were connected with bonds of length b = 5.3 Å (b* = 2) but where next-nearestneighbors were effectively repelled, as described by Rg from the good solventsimulation was reproduced. This procedure resulted in a stiffnessstrength of α = 5.A previously establishedsimulation model for PEG in water at roomtemperatureT* = kT/ϵ, corresponding to our threeexperimentally investigated temperatures: 20, 45, and 75 °C (roughly).We emphasize that one must keep in mind the “inverted”relation; that is, a high “real” temperaturecorresponds to a high fraction of hydrophobic B typemonomers which attract each other, which our model is captured bya low reduced temperature. It turns out that thetheta temperature (UCT for infinitely long polymers) is TC* = kTC/ϵ = 7.65 for our polymer model, wherethe subscript “C” stands for critical. Since we, inthe experiments, are investigating the regime of 20–75 °C(293–348 K), that is, below the LCST, we must in our modelfocus on reduced temperatures above 7.65. One wayto establish model temperatures that “correspond” toexperimental ones  is to borrow results fromprevious efforts, arriving at the approximate temperature dependenceof an “effective” Flory–Huggins (FH) parameter,χeff. The parameter is denoted “effective”as it is temperature-dependent, in contrast to the corresponding parameterin standard FH theory, and thus incorporates entropic effects. Thistype of FH approach is very much in the same spirit as our currentmodeling of chains composed of L-J monomers, with temperature-dependentinteractions. From Figure 11 in the work by Dormidontova,28 we can make crude estimates of χeff for our three target temperatures. In a commonly adopted simplifiedapproach, the FH “energy” of mixing depends on temperature,Δϵmix(T), which of courseimplies that this should be regarded as a free energy parameter. Borrowingconcepts from FH theory, we note that χeff is proportionalto Δϵmix(T)/T. This means that if we assume that our L-J “energy”(free energy) parameters, ϵ(T), scale withtemperature in the same manner as Δϵmix(T), we haveeff(T) to our reduced temperatures:eff(TC) = 0.5, and we recall thatour mean-field theory predicts TC* = 7.65. Hence, we arrive at thefollowing mapping between experimental temperatures and reduced temperaturesin our L-J model, where the values of χeff(T) are estimated from Figure 11 in ref (eff(293 K) ≈ 0.425 = > T* ≈ 9.020 °C: χeff(318 K) ≈ 0.46 = > T* ≈ 8.345 °C: χeff(348 K) ≈ 0.49 = > T* ≈ 7.875 °C: χExtensions of FH theory have been proposed wherethe interactionparameter varies with composition, but these will not be consideredin this work. In order to complete our model, we need to address theinteraction between a PEG monomer and the bare PS surface. We shalldenote this interaction by vPS(z) and assume that it only varies with the separation, z, between the monomers and the PS surface, which is flatin our model. From our previous studies,29 including our A/B model, we anticipatea short-ranged hydrophobic attraction between the PEG monomers andthe bare PS surface. We also expect this attraction to grow strongeras we increase temperature , as the fractionof B monomers increases. We have chosen to model thisattraction by a short-ranged truncated and shifted harmonic potential,acting on all monomers, with a minimum at the (infinitely hard) PSsurface:zc is setequal to one bond length, b, ensuring a short-rangedinteraction. A thus regulates the strength of theattraction. Note that the relevant strength at a given temperatureis measured as A/(kT), that is,how strong the attraction is relative to thermal energy. Thus, witha fixed value of A, we will in practice obtain astronger attraction at “75 °C” (T* = 7.8) than at “20 °C” (T*= 9.0), as A is larger compared to kT in the former case (7.8 < 9.0). This makes sense given our underlyingPEG model, where we anticipate a higher fraction of hydrophobic monomersat higher temperatures.We still need some way to estimatesuitable model values for theenergy L-J parameter ϵ or, equivalently, suitable reduced temperatures, z = 0 and z = h. The total potential, VPS from these surfaces is found by superposition: VPS = vPS(z) + vPS(h – z).Our model slit geometry is confinedby two “PS surfaces”,located at A. Fortunately, our experimentalobservations, in combination with DFT predictions, allows us to arriveat a rather specific value of this quantity. As we shall see, thiswill be further corroborated by agreements between independent predictionsand experimental results for a range of different conditions. We will,however, postpone the estimation of A until the experimentalresults have been reported (below).Armedwith these assumptions and definitions, we need a way toestimate the amplitude 39 Note that there are two different kinds of polymers in our system:the end-grafted chains, which are short with orientationally flexiblebonds, and the dissolved, long, semiflexible polymers.Here, we give a briefaccount of the polymer DFT that was used to calculate interactionfree energies. We refer to earlier work for details.R = {r1,.....,rM} denote the monomer coordinates of a “free”(dissolved) M-mer chain . Furthermore, we will by N(R) denote a multipoint polymer density, from which the densities ofseparate monomer sites, ni, can be evaluated:r) is the Diracdelta function. We also introduce an analogous notation for the graftedchains, which are identified by an index “g”.We let Mg-mers). The chemical potential is calculated as a derivativeof the bulk free energy at the chosen bulk polymer density. Thesepolymers are also under the influence of an external field, VPS, that originates from our planar PS surfacesat z = 0 and z = h. The grand potential, Ω, can be written asFex[{ni(r)}{ni(g)(r)}], originating from all monomer–monomer interactions, can be expressed in terms ofthe site densities, ni(r) (dissolved chains) and ni(g)(r) (surface-tethered chains). These interactions aregenerally decomposed into separate excluded volume (hard core) andlong-ranged attractive parts. In this work, we have managed the formerby utilizing the generalized Flory dimer approximation,35 while the latter is expressed by the attractive part of the L-Jinteraction. Neighboring monomers along the chain are connected bya bonding potential, Φ(b)(R), that isdefined so that the bond distance is fixed at a constant value b:(b)(Rg) (wherethe number of bondsis limited to Mg – 1 = 44).We note that our model comprises a polymer fluidcomposed of semiflexibledissolved chains recall 6 in chemx, y, leaving only z-dependent quantities. We also note that, for our system,the external potential originates from the PS surfaces, modeled asplanar walls, with which the monomers interact; see g[n1(g)(r)] is a Lagrange multiplier, acting on the grafted monomer,which we define as “monomer 1”. Ψg[n1(g)(r)] is adjusted so that the desired grafting densityis achieved. Monomer 1 is in all grafted chains bonded to one of thesurfaces but is allowed to move within one bond length. We formulatethese conditions asg resemblesthe Donnan potential in ionic solutions, which is regulatedin a similar manner to achieve electroneutrality.39In our planar slit geometry, things are simplified by the factthat we can integrate out, in a mean-field manner, all dependenciesupon 39 This may seem like a formidabletask, but very efficient computational approaches have been developed39 that are based on recursive relations. Calculations at a given separationand for a given choice of μ (and thus bulk density) are initiatedfrom an estimate of the equilibrium density profiles for monomersbelonging to free and grafted chains. An initial educated guess ofΨg[n1(g)(r)] is also required.The grand potential minimization leads to integral equations thatin turn generates new predictions for the monomer density profiles,as well as Ψg[n1(g)(r)]. These integralequations are solved recursively so that all possible conformationsof dissolved and grafted chains are accounted for, subject to a z-dependent Boltzmann weight. This weight has a mean-fieldcharacter, although excluded volume correlations, with a range ofabout a monomer diameter, are approximately taken into account. Thebond angle potential, EB, also introducescorrelations between next-nearest neighbors along the chain. In principle,these newly predicted profiles could serve as new guesses for thenext iteration. However, such an approach will in most cases leadto divergence, and a more conservative scheme is recommended whereonly a small fraction of the newly predicted profiles are mixed withthe previous profile. This procedure is sometimes referred to as Picarditerations. The iterations continue until self-consistency is achieved.The equilibrium monomer distribution is found byminimizing thegrand potential using iterative procedures until self-consistencyis obtained.eq/S, for a range of different separations, h, we arrive at a complete free energy curve, which is directlyrelated to the force, F, between two identical particlesof radius Rp via the DA:32F(h) points, followed by yet another integration,generates the final particle–particle PMF, W:By calculating the minimized grand potential per unit area, Ωeq/∂h, agrees with its discrete correspondence,−ΔΩeq/Δh. Suchtests are not provided here but are obviously available upon request.A useful and very sensitive test ofthe written DFT code is thatthe calculated analytical expression for the normal pressure, −∂ΩParticle characterization and thermalanalysis of dispersions werecarried out using DLS and CLSM analysis. We summarize the resultsin the following sections.Accordingto our DLS measurements,the zeta-potential of the PEGylated PS particles in the absence ofany added free PEG was about −3.6 mV, and the ionic strengthwas also quite low at about 0.06 mM. The DLS data furthermore impliedan average particle diameter of 1900 Å, which is slightly smallerthan the value obtained from CLSM analyses: 2300 Å.g2, of a solution containing particles at a concentration of0.05 wt %, as well as PEG with a molecular weight of 100 kDa and aconcentration of 6 mg/mL, was measured at room temperature. The resultsare displayed in 2/s after 48 h. This clearly indicates a transitionfrom fast singular particle movement to slow movements of clusters.Fits to DLSwas also employed to analyze the evolution of aggregates in dispersionswith added PEG. The autocorrelation function, Supporting Information).A brief note here, based on our observations, is that initial aggregatestended to be rather linear, with substantial branching being developedat later stages.In order to shed more light on particle evolution, CLSM was alsoemployed, which will be discussed in more detail below  was then added, arriving at polymer concentrationsin the range of 0.7 to 9 mg/mL. The samples were allowed to equilibrateat various temperatures (20–87 °C) and subsequently imagedby a CLSM. The equilibration time varied depending on both temperatureand concentration of added PEG. For instance, samples at low temperaturesequilibrated on a time scale of 1 day to 2 weeks, depending on thePEG concentration, whereas at higher temperatures, equilibration wasfast and the results were extracted within a short period of 1–3h. Within the intermediate temperature regime, the samples were equilibratedfor 2 weeks. CLSM snapshots are collected in 19 Second, if the attraction was caused by bridgingat room temperature, then it would become monotonically stronger with temperature, as the PEG monomers become progressively morehydrophobic. However, as we shall demonstrate below, there is an intermediatetemperature regime (above ambient) where redispersion to a homogeneoussample takes place, clearly suggesting less attractiveinteractions.At room temperature,the dissolved polymers generate attractive depletion forces. Thereare at least two strong arguments in favor of this conclusion. First,we note that if there was any significant attraction between PEG monomersat room temperature, one would expect gel formation at high particlevolume fraction, even in the absence of added polymer. However, thesedispersions will only form gel at higher temperatures.40Note that a simplifying aspect of these systemsis that the presenceof a grafted PEG layer will allow us to neglect Hamaker interactionsbetween the PS particles.Our findingsfrom CLSM studies, combined with diffusion analyses,were collected at various particle and polymer concentrations, andthe results are summarized in monomer concentrationwas then required to induce depletion flocculation at room temperature.For 2 kDa chains, this corresponds to polymer concentrations thatare several thousand times higher than when 100 kDa PEG was used.For these short polymers, we did not detect any obvious structuralfeatures of clusters formed, as macroscopic phase separation and furthersedimentation were quite rapid.We briefly mention that we also made some tests where the addedPEG chains were quite short, with molecular weights of 2 and 8 kDa.A 40–50 times higher Tr = 20 to T = 50, 73, and 78 °C,as illustrated by a heating ramp in Tr immediatelyafter polymer addition until they reach equilibrium or were heatedto higher T immediately after PEG addition and thenequilibrated before imaging. An additional experiment was performedon particles which aggregated at low Tr and were heated to a target temperature, T, afteraggregation. As shown in The measurementsthat we have reported so far were recorded at room temperature. However,in order to evaluate our theoretical predictions, we performed a thermalanalysis on the dispersions at temperatures ranging from A further temperatureincrease to about 70–80 °C leadsto a pronounced formation of branched clusters.only contain PEG-grafted PS particles. We expect that clusters mightform in such systems at high enough temperatures, as the PEG layersthen become hydrophobic. On the other hand, this is merely a qualitativeargument, and it is clearly of interest to establish quantitativeresults which we subsequently can compare (see below) with predictionsfrom our theoretical model. We have summarized our analyses for thesystems in We brieflyreport CLSM analyses on structures of dispersions that 29 has demonstratedthat in the intermediate regime, that is, for intermediatelyadsorbing surfaces, the polymer-mediated interaction is repulsive at equilibrium. These predictions were admittedly based on approximateDFT, but for the special case of ideal chains composed of bonded point-likemonomers, DFT results are exact, and the predictionswere shown to be valid also for those model systems. We provide furtheranalyses of such systems in the A, so that it leads to a net repulsive interaction (intermediate adsorption) at T* = 8.3, which we recall is our model representation ofthe 45 °C experimental system.Experimentally, we observe a redispersionof clusters and flocculated structures at roughly 45 °C. Thismay seem surprising, given the well-established mechanisms accordingto which equilibrium interactions mediated by dissolved polymers areattractive between nonadsorbing surfaces, due to depletion, but alsoattractive between adsorbing surfaces via bridging interactions. However,we reiterate that recent workA-scan, searching for a value that leads to a polymer-inducedrepulsion at T* = 8.3 (“45 °C”).In A can be quite dramatic! Across the parameter regime, A = 2.25 → 2.55, the model PS surface effectively changes frombeing nonadsorbing to adsorbing. This is illustrated in graph (b)of h = 200σ. In agreementwith results in ref . In other words, we use the observation of redispersionat our intermediate temperature as a way to measure the “proper”strength of the hydrophobic attraction. With A =2.4, the maximum strength of the surface potential, vPS(z = 0), is, at T*= 8.3, slightly larger than thermal energy: βvPS(z = 0) ≈ −1.16.Now we proceed with our s in ref , the polNow that we have setall model parameters,based on previous works on PEG+water solutions, and our observationsof redispersion of our PEG-grafted PS particles at 45 °C, weare ready to make theoretical predictions for PMFs and concomitantstructures at higher and lower temperatures (with 45 °C as ourreference).T* = 7.8) system. As we shall see below,this originates from interactions between the grafted layers, whichdisplays an attractive regime at this and higher  temperatures.Calculated polymer-mediated PMFs at our model representationof 20, 45, and 75 °C are collected in Using ourestablished model, we can of course also predict PMFsbetween our grafted particles in the absence of any added polymer.The polymer-induced interactions will then only be due to the tetheredchains. At low temperatures, this will lead to a monotonic repulsion,as the entropic cost of reducing the number of available configurationsas another surface approach will dominate the weak L-J attractionsbetween the monomers. However, at high temperatures, the latter willprovide an attractive regime.T* = 7.8 in our model L-Jsystem. Hence, if we wish to study a temperature regime very closeto this value, say, 70–78 °C, it makes sense to utilizea simple linear scaling around this value so that 70 °C correspondsto a model reduced temperature of T* = 7.8 ×348 K/343 K ≈ 7.91, while T* = 7.8 ×351 K/348 K ≈ 7.73 provides a model representation of 78 °C.Calculated PMFs at these temperatures are shown in T*= 7.8 curve in According to experiments, clusterswill form in samples free of added polymers, at 78 °C, but thisis indeed close to the “transition point”; that is,they will not form at temperatures substantially lower than this thresholdvalues. It is obviously of interest to see what our theoretical modelpredicts at similar temperatures. We have already established a temperaturemapping according to which 75 °C “corresponds to”a reduced temperature of 41 has demonstrated that, in the presenceof such a double-layer barrier, the width of the adhesive minimumhas a strong impact on the structure of the clusters. Specifically,a narrow attraction leads to more linear clusters, which is exactlywhat we observe in this work.There is another aspect worth noting. According to our CLSM analyses,the clusters that are formed at about 78 °C in the absence ofadded PEG tend to be more linear than when the dissolved polymersmediate the attraction. This is in agreement with the prediction aboveof a substantially more short-ranged attraction in samples free ofdissolved polymers. Note that we have not included an expected weakbut long-ranged electrostatic repulsions, given that DLS data suggesta weak particle charge and a low ionic strength. Recent simulationwork29 wepredicted that our experimental system would display a non-monotonictemperature dependence where flocculation at high and low temperatures(for a sufficiently high dosage of added polymer) is replaced by afully homogeneous dispersion at some intermediate temperature. Thisqualitative prediction was indeed confirmed by our experimental investigations.Using the experimental data, we have constructed a designated (specific)coarse-grained model for this particular system, whereby semiquantitativeagreements can be achieved using a minimum number of parameters. Weemphasize that all our measurements and calculations were performedat supracritical conditions, which experimentally corresponds to temperaturesconsiderably below the bulk LCST. Hence, we do not expect any polymerconfigurational transitions, such as a coil to globule transition.Basedon previous theoretical modeling,We hope that our work may pave the way for new options to regulatecolloidal stability, where polymer addition is combined with changesof solvent quality using (for instance) pH, salt, or temperature control.


To (i) determine whether sustained disease activity states, as measured by Bath Ankylosing Spondylitis Disease Activity Index (BASDAI) and Ankylosing Spondylitis Disease Activity Score (ASDAS), impact function, and (ii) evaluate characteristics predicting sustained low functional impairment in a prospective axial spondyloarthritis (axSpA) cohort.Biologic Treatment Registry Across Canada (BioTRAC) was a multi-center, prospective registry that collected real-world data on axSpA patients receiving infliximab or golimumab between 2006 and 2017. Generalized estimating equations (GEE) were used to test baseline characteristics, treatment, and duration (at 6 and 12 months vs. only at 6 or 12 months vs. neither) of low BASDAI (< 3), ASDAS-inactive disease (ID)(< 1.3), and ASDAS-low disease activity (LDA) in predicting sustained low Bath Ankylosing Spondylitis Functional Index (BASFI)(< 3) between 12 and 18 months. The adjusted impact of achieving low disease state at 6 and/or 12 months on BASFI at 18 months was analyzed by generalized linear models.p < 0.001), low BASDAI only at 6 or 12 months (p = 0.001), and baseline BASFI (p < 0.001) were the only predictors of sustained low BASFI. Sustained ASDAS-ID (p = 0.040) and ASDAS-LDA (p < 0.001) were also predictors when forced into the model. Similar results were obtained when evaluating the BASFI score at 18 months.Eight hundred ten patients were enrolled. 33.7%, 13.4%, and 24.7% achieved sustained low BASDAI, ASDAS-ID, and ASDAS-LDA, respectively. In univariable GEE of baseline variables, age and baseline BASDAI, BASFI, and ASDAS significantly predicted sustained low BASFI. In multivariable GEE, sustained low BASDAI (Sustained BASDAI < 3 may be a valid and feasible target for a treat-to-target strategy in axSpA having function as treatment goal.The online version contains supplementary material available at 10.1186/s13075-023-03055-1. Axial spondyloarthritis (axSpA), comprising ankylosing spondylitis (AS) and non-radiographic axSpA, is a chronic inflammatory rheumatic disease affecting the axial skeleton , 2. AlthCurrent international recommendations for the management of axSpA call for the use of non-steroidal anti-inflammatory drugs (NSAIDs) as a first-line treatment followed by anti-tumor necrosis factor (TNF) agents and, more recently, interleukin (IL)-17A inhibitors , 8, 9, wThere is also no consensus as to which outcome(s) should be monitored/targeted in a T2T strategy, though there is agreement that the key domain is disease activity , 14. TheThere is also the question whether maintenance of a low disease activity state over a prolonged period in axSpA is desirable, as documented for rheumatoid arthritis, where sustained low disease activity over at least 6 months is associated with improved functional and structural damage outcomes .The objectives of the current study were to determine whether and to what degree sustained (≥ 6 months) low disease activity states, as measured by BASDAI < 3 and ASDAS (< 1.3 and < 2.1), impact function and which of these states was most impactful. We also aimed to explore patient and disease characteristics as predictors of sustained low functional impairment, focusing on a comparison of sustained low ASDAS, both ASDAS-ID and ASDAS-LDA, versus low BASDAI.The Biologic Treatment Registry Across Canada   was a muIn order to be eligible for BioTRAC, patients needed to have a rheumatologist diagnosis of axSpA, be either bio-naïve (2006) or with ≤ 1 prior biologic agent exposure (2006–2017) and be eligible to receive either infliximab or golimumab as per the Canadian Product Monographs , 22. No The following patient-reported, clinical, and laboratory outcomes were collected, per routine care, in the BioTRAC registry at baseline and every six months thereafter: AS Disease Activity Score (ASDAS), Bath AS disease activity index (BASDAI), Bath AS functional index (BASFI), health assessment questionnaire (HAQ), patient global assessment (PtGA), physician global assessment (MDGA), back pain, enthesitis  , dactyliand 12 months, at only 6 or 12 months, or at neither time; (ii) ASDAS-inactive disease (ID) (< 1.3) at 6 and 12 months, at only 6 or 12 months, or at neither time; and (iii) ASDAS low disease activity (LDA) (< 2.1) at 6 and 12 months, at only 6 or 12 months, or at neither time.For the purposes of this analysis, the main endpoints of interest were (i) BASFI at 18 months of treatment and (ii) low BASFI (< 3) at 12 and 18 months. The main exposures of interest were achievement of (i) low BASDAI (< 3) at 6 p < 0.1) in the univariable analysis were entered in multivariable GEE models; stepwise backward variable selection was used to derive a parsimonious model. The impact of achieving sustained low BASDAI or ASDAS-ID at 6 and/or 12 months on the BASFI score at 18 months were analyzed using generalized linear models adjusted for age, gender, baseline disease duration, and baseline BASFI score. An additional model was built where the interaction of normal CRP (< 5 mg/L) with low BASDAI at 6 and/or 12 months was included as a predictor of BASFI at 18 months. For both the GEE and generalized linear models mentioned above, a secondary analysis was run in which ASDAS-ID was replaced with ASDAS-LDA.Baseline characteristics and disease parameters were summarized using descriptive statistics, namely the mean and standard deviation for continuous variables and counts and proportions for categorical variables. Univariable generalized estimating equations (GEE) were used to test the impact of baseline age, disease duration, gender, smoking, enrollment period, prior biologic experience, anti-TNF agent, non-biologic disease-modifying anti-rheumatic drug (nbDMARD) use, NSAID use, steroid use, presence of enthesitis and dactylitis, CRP, ASDAS, BASDAI, and BASFI and follow-up  parameters on achievement of sustained low BASFI at 12 and 18 months. Parameters showing a statistical trend .A total of 810 patients were included, with a mean  age of 45.9 (SD 12.7) years and disease duration of 7.4 (10.1) years , ASDAS score , BASDAI score , and BASFI score  were identified as negative predictors of sustained low BASFI (< 3) between 12 and 18 months. In contrast, treatment with golimumab  and achievement of low BASDAI, ASDAS-ID, and ASDAS-LDA at 6 and/or 12 months were positive predictors of sustained low BASFI (< 3) between 12 and 18 months  or at either 6 or 12 months (but no sustained), were the only significant predictors of low BASFI (< 3) between 12 and 18 months , baseline BASFI score , and low BASDAI, both sustained  and at either 6 or 12 months , were strong predictors of BASFI score at 18 months, while sustained ASDAS-ID was a weaker predictor (Table P = 0.046) by which sustained normal CRP was identified as an independent predictor of improved function among patients with sustained low BASDAI . No other significant interactions were observed.Inclusion in the model of the interaction term low BASDAI by normal CRP (< 5 mg/L) at 6 and 12 months, showed a significant interaction  concurrently. Nonetheless, given that calculation of BASDAI does not include acute phase reactants and, thus, is more feasible in routine clinical care, this is an important finding that may have a significant impact on the application of T2T in daily practice. Achievement of sustained normal CRP among patients in sustained low BASDAI was found to be associated with further improvement in function, suggesting the importance of further lowering inflammation in these patients. This incremental benefit of CRP over and above low BASDAI may also be true for patients not achieving low BASDAI; however, this was not possible to evaluate due to the low number of patients that were not in sustained low BASDAI but were in sustained normal CRP.Various cut-offs for defining a low disease activity state according to the BASDAI have been proposed such as < 2, < 3, and < 4. Similar cut-offs have been proposed for what constitutes an acceptable functional state. Some cut-offs have been validated against an external reference, this being most commonly the patient acceptable symptom state (PASS) question which asks patients whether they would globally consider their condition acceptable if they were to remain in their present state for the next month or the rest of their life , 29, 30.To our knowledge, this is one of the first studies directly comparing clinical measures of disease activity to determine which measures could be used for target monitoring in a T2T strategy in axSpA. In a previous pragmatic prospective, one-year trial, intensified treatment strategy aiming to achieve ASDAS < 2.1 was not associated with a significant benefit in terms of achieving a ≥ 30% improvement on the Assessment of SpondyloArthritis International Society (ASAS) Health Index (HI), the primary endpoint, compared to standard of care ; furtherOur study has certain limitations. The study was not designed to use a T2T protocol, even though T2T may have been used in routine care by some rheumatologists, or to be sufficiently powered to identify predictors with small to moderate effects which may have resulted in the non-detection of certain associations. Moreover, analysis of sustained endpoint achievement is limited in the context of a 4-visit, 18-month study. The analysis also does not include measures of Ankylosing Spondylitis Quality of Life (ASQoL) or ASAS HI. Furthermore, patient loss to follow-up may have resulted in selection bias, and radiographic progression was not evaluated. In addition, although the relative value of monitoring BASDAI and ASDAS was evaluated, a comprehensive assessment of other composite indices was not conducted; additional future studies will be required to evaluate the usefulness of other targets. Finally, although disease duration at baseline was not identified as a significant predictor of function, the findings may not be generalizable to patients with early disease.Aiming for sustained low BASDAI (< 3) may be a valid and feasible T2T treatment strategy for routine care in axSpA. Further validation of these real-world findings is required to aid in achieving consensus on which outcome should be monitored in a T2T strategy.Additional file 1:Supplementary Table 1. Predictors of Sustained Low BASFI (<3) Between 12 and 18 Months Using Non-Responder Imputation . BASFI=Bath Ankylosing Spondylitis Functional Index; OR=Odds Ratio; CI=Confidence Interval; M=Month; BASDAI= Bath Ankylosing Spondylitis Disease Activity Index; ASDAS=Ankylosing Spondylitis Disease Activity Score; ID=Inactive Disease; LDA=Low Disease Activity. Bold P-values denote statistical significance. Multivariable Analysis 1: primary analysis; Multivariable Analysis 2: secondary analysis including ASDAS-LDA instead of ASDAS-ID.


The prevalence of type 2 diabetes mellitus (T2DM) is increasing each year and has become one of the most prominent health concerns worldwide. Patients with T2DM are prone to infectious diseases, and urinary tract infections are also widespread. Despite a comprehensive understanding of urinary tract infection (UTI), there is a lack of research regarding primary prevention strategies for asymptomatic bacteriuria (ASB).To clarify the incidence and risk factors of asymptomatic urinary tract infection in patients with T2DM by meta-analysis to provide evidence for preventing UTI. Help patients, their families, and caregivers to identify the risk factors of patients in time and intervene to reduce the incidence of ASB in patients with T2DM. Fill in the gaps in existing research.Meta-analyses were conducted in line with PRISMA guidelines.Eleven databases were systematically searched for articles about ASB in T2DM, and the retrieval time was selected from the establishment of the database to February 5, 2023. Literature screening, quality evaluation, and meta-analysis were independently performed by two researchers according to the inclusion and exclusion criteria, and a meta-analysis was performed using Stata 17.0.P < 0.001). After controlling for confounding variables, the following risk factors were associated with ASB in patients with T2DM: age , I2 = 75.5%, P < 0.001), female sex , I2 = 79.3%, P = 0.002), duration of type 2 diabetes , I2 = 80.7%, P < 0.001), HbA1c , I2 = 62.6,%. P < 0.001), hypertension , I2 = 0%, <0.001), hyperlipidemia , I2 = 0%, P < 0.001), Neuropathy , I2 = 0%, P < 0.001), proteinuria , I2 = 62.7%, P < 0.001).Fourteen articles were included, including cohort and case–control studies. A meta-analysis of 4044 patients with T2DM was included. The incidence of ASB in patients with T2DM was 23.7%; The overall prevalence of ASB in T2DM is 23.7%. Age, female sex, course of T2DM, HbA1C, hypertension, hyperlipidemia, neuropathy, and proteinuria were identified as related risk factors for ASB in T2DM. These findings can provide a robust theoretical basis for preventing and managing ASB in T2DM. Type 2 diabetes mellitus (T2DM) is a metabolic disease characterized by hyperglycemia caused by insulin resistance or insufficient insulin secretion. Clinical features include polydipsia, polyuria, and weight loss , 2. LongASB patients have no symptoms or signs of urinary tract infection but are positive after repeated urine bacterial cultures; in these patients, the number of colonies after two consecutive cultures reaches 105/ml . CurrentEleven databases, including PubMed, Web of Science, Cochrane, Embase, Ovid, Scoups, Chinese full-text journal database, VIP database, Wan Fang database, The National Medical Journal of China, and Sinomed database, were queried, and Endnote X9 software was used to screen the standard literature. Stata 17.0 software was used to perform a meta-analysis of the included literature. The retrieval period was from the establishment of the database to February 5, 2023. The retrieval strategy adopted a combination of subject and free words. For data retrieval from Chinese databases, we used the keywords “asymptomatic bacteriuria,” “occult bacteriuria,” “bacteriuria,” “pyuria,” “Leukocyturia,” “asymptomatic urinary tract infection,” “occult urinary tract infection,” “asymptomatic bladder infection,” “occult bladder infection,” “asymptomatic ureteral infection,” “occult ureteral infection” and “type 2 diabetes mellitus.” The retrieval strategy for the English database is shown in the Patients: (1) Type 2 diabetes mellitus with asymptomatic bacteriuria diagnosed definitively; (2) age ≥18 years; and (1) Case-control study or cohort study. (2) The language of the literature is Chinese or English.(1) Abnormal urinary system structure; (2) Document types such as conference reports, case reports, and reviews; (3) inability to obtain complete literature, repeated publications, or incorrect statistical methods; (4) Newcastle Ottawa Scale Score (NOS) < 6.After importing articles into Endnote X9, the titles and abstracts of the articles were independently read and screened by two researchers (MengQiao Dai and Shan Hua) who had systematically studied evidence-based courses. The remaining documents were independently read and screened according to the exclusion criteria. The two researchers independently extracted data from the literature that met the inclusion criteria, including author information, publication year, risk factors, literature type, sample size, age, prevalence rate, and related risk factors. While selecting articles and extracting information, if two researchers disagreed, a third researcher with the same qualification was consulted to reach a consensus to ensure the quality of the included articles.Two researchers independently evaluated the quality of the articles and scored them using the NOS scale. The NOS score includes three large blocks, with eight items to evaluate case-control or cohort studies, including the selection, comparability, exposure, evaluation, and results of the research population. The total score was 9, and articles with a score of less than 6 were excluded. If two researchers disagreed on the quality evaluation of an article, a third researcher with the same qualification was consulted to reach a consensus.P ≥ 0.1 and I2 ≤ 50% suggested that the study was homogeneous, the fixed effect model was adopted. Studies were considered heterogeneous if the heterogeneity was P < 0.1 and I2 > 50%. The random effects model was used to determine the source of heterogeneity through sensitivity analysis or subgroup analysis. If the heterogeneity was still large, a random-effect model combined with a descriptive analysis was used to express the results. According to the Cochrane website recommendations, a funnel chart was used for publication bias analysis when a single risk factor was present in more than ten articles.Stata 17.0  was used to analyze the data. The odds ratio (OR) was used as the effect index for counting data, the weighted mean difference (WMD) was used as the effect index for measuring data. The interval estimation was expressed as a 95% confidence interval. If the heterogeneity test showed A total of 1668 articles were retrieved; 314 duplicates were removed using Endnote X9 software, and the remaining 1354 articles were used for further analysis. After two researchers read the titles and abstracts of the articles, 1307 articles were excluded, leaving 47 documents. After reading the specific contents of the literature according to the exclusion and inclusion criteria, 14 studies were included in the meta-analysis. A flow diagram of the search and selection of studies is shown in Fig. Fourteen articles –27 were Literature quality evaluation was scored using the NOS scale, with 9 being the highest score and greater than or equal to 6 being a good-quality cohort study or case-control study. In the scoring results, All case-control studies meet the requirements of “Adequate definition of cases,” “Representativeness of the cases,” The conditions of “ascent of exposure,” and “same method of ascent for cases and controls” are not met by some cohort studies. Please refer to Table P < 0.001) Fig. A meta-analysis identified 910 patients with T2DM complicated with ASB and 3494 patients without asymptomatic bacteriuria. The incidence of ASB in T2DM was 23.7%  Fig. The 14 articles  includedI2 = 79.3%, P = 0.002)  Fig. . Owing tI2 = 80.7%, P < 0.001) Fig. I2 = 84%, P < 0.001), and the primary source of heterogeneity was foreign articles  Fig. The included 14 articles  includedI2 = 62.6%, P < 0.001)  Fig. . We cond01) Fig. . The funI2 = 84.9%, P = 0.359)  Fig. .Fig. 12MI2 = 75.3%, P = 0.803)  Fig. .Fig. 13MI2 = 0%, P < 0.001)  Fig. .Fig. 14MI2 = 0%, P < 0.001)  Fig. .Fig. 15MI2 = 0%, P < 0.001)  Fig. .Fig. 16MI2 = 62.7%, P < 0.001)  Fig. .Fig. 17MFourteen studies included in this meta-analysis showed that the incidence of ASB in patients with T2DM was 23.7%, higher than that reported by Guide  and ZhanAge was a risk factor for ASB. The decline in self-function in such patients, coupled with diabetes, leads to an imbalance in the internal environment of patients . The metFor patients with these characteristics, clinicians should observe the symptoms related to urine and the urinary system. For female patients with T2DM, awareness of self-cleaning should be widely popularized, and behaviors such as urination immediately after sexual activity, changing and cleaning underwear on time, improving immunity, and using immune enhancers appropriately, should be encouraged.The course of the disease and HbA1c are risk factors for ASB in patients with T2DM. Studies have shown that the possibility of ASB infection increases by 1.9 times every 10 years in patients with diabetes . Long-teDiabetic nephropathy (DN) is one of the leading causes of end-stage renal disease worldwide . GlomeruFor patients with diabetes, symptom management is critical so that patients’ blood sugar can reach an ideal level, effectively preventing the emergence of acute and chronic complications of diabetes. The management of DN is not limited to drugs and other treatments; lifestyle interventions are also important measures to promote health. Healthy eating habits, weight control, proper exercise, smoking cessation, and good sleep habits can effectively delay the progression of kidney disease in patients with diabetes.+ efflux or generation of reactive oxygen species, which are considered classical activators of the NLRP3 inflammasome during hypertension -like receptor family pyrin domain containing 3) activation and inflammasome assembly is triggered through either ATP-induced Krtension . Asymptortension , 54. Thertension –57. In prtension . CaregivCross-sectional studies on related topics were not included in this study, resulting in a small number of documents eventually being included; most of the people included in the studies were from developing countries. Therefore, the final meta-analysis results only represent the main risk factors for ASB in some patients with T2DM, and there may be some bias. Due to the differences in the characteristics of research objects, the number of samples, and the methods and instruments used to determine related indicators among different studies, the results of the meta-analysis are relatively heterogeneous. The exact reasons behind the reduced heterogeneity of results remain unclear. Therefore, it is suggested that more high-quality, multi-center, and large-sample original research should be conducted in the relevant direction to provide more targeted measures and generate more scientific evidence-based recommendations for patients with T2DM.There is no evidence that ASB treatment can reduce the incidence of UTI. Therefore, it is vital to pursue prevention as the primary strategy in patients with T2DM . This stAppendix


Neurospora circadian clock, the core element, FREQUENCY (FRQ), binds with FRQ-interacting RNA helicase (FRH) and casein kinase 1 to form the FRQ–FRH complex (FFC) which represses its own expression by interacting with and promoting phosphorylation of its transcriptional activators White Collar-1 (WC-1) and WC-2 . Physical interaction between FFC and WCC is a prerequisite for the repressive phosphorylations, and although the motif on WCC needed for this interaction is known, the reciprocal recognition motif(s) on FRQ remains poorly defined. To address this, we assessed FFC–WCC in a series of frq segmental-deletion mutants, confirming that multiple dispersed regions on FRQ are necessary for its interaction with WCC. Biochemical analysis shows that interaction between FFC and WCC but not within FFC or WCC can be disrupted by high salt, suggesting that electrostatic forces drive the association of the two complexes. As a basic sequence on WC-1 was previously identified as a key motif for WCC–FFC assembly, our mutagenetic analysis targeted negatively charged residues of FRQ, leading to identification of three Asp/Glu clusters in FRQ that are indispensable for FFC–WCC formation. Surprisingly, in several frq Asp/Glu-to-Ala mutants that vastly diminish FFC–WCC interaction, the core clock still oscillates robustly with an essentially wildtype period, indicating that the interaction between the positive and negative elements in the feedback loop is required for the operation of the circadian clock but is not a determinant of the period length.In the negative feedback loop composing the  Neurospora crassa, a circadian model organism used for decades, the White Collar complex (WCC) derived from all cellular White Collar-1 (WC-1) and a fraction of WC-2 drives transcription of the central pacemaker gene, frequency (frq), through binding to either of two DNA elements in the frq promoter under contrasting illumination conditions: either the Clock box (C-box) in the dark or the proximal light-response element upon light exposure or in constant light   , 8, 9 anelicase)  to createlicase) , 11, 12 Neurospora, while much is becoming known about the means through which repression is achieved  and CRYPTOCHROMEs (CRYs) which form a negative element complex with CK1, analogous to the FFC, that inactivates CLOCK/BMAL1 by phosphorylation at key residues , 14. As achieved , 17, 18,Neurospora, WCC acts as a responsive hub coordinating the circadian clock that is obligate in constant darkness and light signaling  and its nearby DBD (defective in DNA binding) motif, and two Per-Arnt-Sim (PAS) domains for WC-2 interaction  period length; frqΔ691–732 displays a shortened period of 15.5 h; frqΔ107–148, frqΔ435–481, frqΔ604–646, frqΔ874–884, and frqΔ857–905 have prolonged rhythms to variable extents. The core clock became totally arrhythmic in frqΔ149–193, frqΔ194–199, frqΔ200–249, frqΔ250–310, frqΔ482–510,frqΔ511–558, frqΔ647–690, frqΔ733–774, frqΔ775–800, and frqΔ801–856. The data support the idea that multiple regions of FRQ take part in the rhythmicity control and period determination, as has been documented by a large body of literature.Three distinct regions on FRQ have been implicated in WCC interaction: amino acids 107 to 310, 435 to 558, and 631 to 905 . The loselements  might beeal-time . frqΔ564frq mutants whose circadian phenotypes were analyzed by the luciferase assay in frq mutants that were grown in the light and tested  in the d tested . When FRactivity  and 2. Fd length  and 2, sthmicity  and 2. TΔ801–856 , which md frq6B5 . This ald frq6B5 , presumascillate , consisteraction . InteresA). Similarly, in a separate experiment with epitope-tagged FRQ instead, FRQV5H6 as well as bound FRH were immunoprecipitated readily with V5 antibody regardless of salt titers, whereas WC-1 and WC-2 disappeared from the FFC when salt dosages rose . The data support that FFC and WCC associate with each other by virtue of electrostatic forces, which are subject to high salt disruptions in vitro, while WC-1 and WC-2 bind together presumably by hydrophobic effects via their PAS domains (A basic stretch (KKKRKRRK) near the ZnF DNA-binding domain of WC-1 was previously identified as required for the WCC–FFC organization , which sFig. 3 domains ; likewis domains , 33. Accfrq D/E-to-A mutants  relative to WT, though FRQ and FRH in these strains interact normally with each other  (A). FRQ expression in frqD773A, D774A is extraordinarily low (a faint band after IP) (B), explaining the arrhythmicity in the luciferase assay (A). WC-1 and WC-2 levels in FFC–WCC became greatly diminished in frqD149A, D150A, once again showing that the WCC–FFC abundance in the cell does not faithfully reflect the period length  is consistent with abolished rhythmicity (A). These data reinforce the necessity of incorporating FFC into WCC for eliciting phosphorylation-induced repression. It is noteworthy that unlike frqE835A in which FRQ protein accumulation was abolished , suggesting that the negative charge of E835 is the main contributor in promoting FRQ accumulation.FRQ was completely undetectable in frqE835A . To probne peak) A. FRQ exfter IP) B, explaise assay A. WC-1 ad length . The losA, D774A B is consthmicity A. These bolished , frqE835 with WT B, suggesfrq mutants each bearing abridged versions of FRQ, and these truncated FRQ segments together cover all the residues of the full-length FRQ twice: frq1–558 and frq559–989, frq1–310, frq311–628, and frq629–989 (A). As expected, frq1–310, frq311–628, and frq629–989 totally lost rhythmicity in the luciferase assay (A). The FRQ level in frq1–310 fell below our detection limit in Western blotting even after an enrichment by IP, while in other mutants, it appears to be comparable to that in WT (B). Although FRH complexed with FRQ as strongly in frq559–989 and frq629–989 as in WT, WC-1 and WC-2 in all of these mutants was undetectable above the background in Western blotting (B), indicating that preservation of individual D/E clusters is not sufficient for FFC–WCC formation.Certain residues on FRQ, including D773, D774, and E835, contribute significantly to the accumulation of the full-length protein  and 6. Wq629–989 A. As expse assay A. The FRat in WT B. Althoublotting B, indicafrqD660E and assayed it for C-box activity by luciferase analysis and WCC interaction by IP. To our surprise, frqD660E behaved as arrhythmically as frqD660A (A) and failed to rescue the loss of the FFC–WCC in frqD660A , are required for WCC association, but surprisingly, frqD664A, D667A, frqD862A, D866A, D867A, D869A, D870A, and frqD874A, D875A, E876A, E877A, E879A, E880A, E882A, E883A, D884A displayed vigorous circadian rhythms with a period length almost identical to WT. These data indicate that the stability of the interaction between the positive and negative elements does not control the pace of the oscillator. It remains elusive whether the FRQ’s D/E residues discovered in this study are located on the interface of the FRQ structure and contact the WCC directly or whether they contribute to the maintenance of FRQ’s tertiary structures that are needed in WCC recognition.In ruitment  and 2. Iruitment . Biochemruitment . In addiruitment . IP assaruitment B, are refrq mutants with a constant WCC-FRQ level showed varying period lengths . This has been suggested by experimental data showing that downregulation of WCC does not greatly perturb circadian rhythmicity beyond a slight ∼2 h period lengthening . Strong defects in FRQ phosphorylation have been noticed in several frq mutants from this study, including frqD773A, D774A (B), frqE760A, E763A, D773A, D774A , and frq311–628 (B), which suggest that CK1 does not function normally in phosphorylating FRQ in these frq mutants given that CK1 is a stable component of FFC and also the major kinase in mediating FRQ phosphorylation (See the paragraph of "FRQ functions mainly as…"). In summary, we confirmed that multiple regions of FRQ are required for FFC to bind WCC and identified important negatively charged residues of FRQ required for WCC interaction.FRQ phosphorylation has been extensively explored for decades as the primary regulatory mechanism governing the ra clock , 49, 50.ra clock  and 6B. A, D774A B, frqE76q311–628 B, which Neurospora strain 661-4a , which serves as WT in luciferase assays, contains the frq C-box promoter fused to the codon-optimized firefly luciferase  at the his-3 locus . To introduce deletions falling in aa 1 to 214 of FRQ, two PCR reactions were performed: one with a forward primer “frq segment 1F” (5′-GAACCAGAACGTAGCAGTGTG-3′) and a reverse primer “del.aa # to # R” bearing a deletion to FRQ and the other using a forward primer “del.aa # to # F” which is reverse and complementary to “del.aa # to # R” and a reverse primer “frq segment 1R” (5′-GACGATGACGACGAATCGTG-3′), and then, the two PCR products were cotransformed into yeast along with pCB05 digested with BstXI  and XhoI  to generate a circular construct. Likewise, for mutations falling in aa 215 to 437 of FRQ, primers “frq segment 2F” (5′-GTGAGTTGGAGGCAACGCTC-3′) and “frq segment 2R”  to NruI ; “frq segment 3F” (5′-GTCGCACTGGTAACAACACCTC-3′) and “frq segment 3R” (5′-CAGCACATGTTCAACTTCATCAC-3′) were designed for pCB05 digested with NruI  and FseI  (FRQ aa 438–675), and “frq segment 4F” (5′-CACCGATCTTTCAGGAGACCCTG-3′) and “frq segment 4R” (5′-CACTCAGGTC TCAATGGTGA TG-3′) pair with pCB05 digested with FseI  and MluI  (FRQ aa 676–989). If deleting regions or mutating residues involve two or more segments above, corresponding restriction enzymes and primers encompassing the regions were selected and combined for yeast recombination. All targeted mutations were validated by cycle sequencing with frq-specific primers at the Dartmouth Core facility. All these frq variants were targeted for homologous recombination at its native locus. Plasmids verified by Sanger sequencing were linearized with AseI  and SspI  and purified with the QIAquick PCR Purification Kit  for Neurospora transformation. Neurospora transformation via electroporation  was performed using an electroporator  as previously reported  with cOmplete, Mini, EDTA-free Protease Inhibitor Cocktail  was added to the powder, and the mixture of the Neurospora tissue powder and buffer was treated with cycles of vortexing for 10 s and resting on ice for another 10 s for a total of 2 min. For WB, equal amounts (15 μg) of centrifugation -cleared whole-cell lysate were loaded per lane in a commercial 3 to 8% 1.5-mm × 15-well Tris-Acetate SDS gel  with 1× NuPAGE Tris-Acetate SDS Running Buffer . Rabbit V5 antibody , mouse FLAG antibody , or rabbit HA  was diluted at 1:5000 as the primary antibodies (Protein lysates for Western blots (WBs) were prepared as previously described , 57. Liqtibodies . Custom tibodies , 58.Neurospora lysate was performed as previously described  containing the frq C-box fused to the codon-optimized firefly luciferase gene  at the his-3 locus.Luciferase assays were performed as previously described , 60. 96-Neurospora strains made in this study are available upon request. All data used to draw conclusions of the article have been presented within the figures.The The authors declare that they have no conflicts of interest with the contents of this article.


His-Purkinje system pacing (HPSP), including his-bundle pacing (HBP) and left bundle branch area pacing (LBBaP), imitates the natural conduction of the heart as an alternative to biventricular pacing (BVP) in cardiac resynchronization therapy (CRT). However, the feasibility and efficacy of HPSP were currently only evidenced by studies with a limited sample size, so this study aimed to provide a comprehensive assessment through a systematic review and meta-analysis.In order to compare the clinical outcomes associated with HPSP and BVP in patients for CRT, PubMed, EMBASE, Cochrane Library and Web of Science database were searched from inception to April 10, 2023. Clinical outcomes of interest including QRS duration (QRSd), left ventricular (LV) function and New York Heart Association (NYHA) functional classification, pacing threshold, echocardiographic and clinical response, hospitalization rate of HF and all-cause mortality were also extracted and summarized for meta-analysis.P < 0.001, I2 = 91%], greater LV functional improvement with increased left ventricular ejection fraction (LVEF) , decreased left ventricular end-diastolic dimension (LVEDD) , and more improved NYHA functional classification . In addition, HPSP was more likely to have higher echocardiographic , clinical  and super clinical  responses than BVP, and a lower hospitalization rate of HF , while presented no difference  in all-cause mortality compared with BVP. With threshold change taking into account, BVP was less stable than LBBaP , but had no difference with HBP .A total of 13 studies  involving 1,121 patients were finally included. The patients were followed up for 6–27 months. Compared with BVP, CRT patients treated by HPSP presented shorter QRSd [mean difference (MD): -26.23 ms, 95% confidence interval (CI): -34.54 to -17.92, The present findings suggested that HPSP was associated with greater improvement of cardiac function in patients with indication for CRT and was a potential alternative to BVP to achieve physiological pacing through native his-purkinje system.The online version contains supplementary material available at 10.1186/s12872-023-03307-7. Cardiac resynchronization therapy (CRT) can effectively improve the symptoms of patients with heart failure (HF) and cardiac electromechanical dyssynchrony. A large number of randomized clinical trials have demonstrated the efficiency of biventricular pacing (BVP) in improving left ventricular (LV) function as well as New York Heart Association (NYHA) functional classification, and reducing rehospitalization in patients with HF and cardiac conduction block –6. HowevSubsequently, new physiological pacing method, i.e., his-purkinje system pacing (HPSP), was developed gradually, which included his-bundle pacing (HBP) and left bundle branch pacing area (LBBaP). In 2000, Deshmukh et al. first reported the HBP activated intrinsic conduction system through the his-purkinje system . Then, LBVP still remains first-line therapy in view of the substantial evidence of efficacy and safety. However, HPSP is very likely to play a growingly important role in the future, and the current recommendations may need to be revised if future long-term evidence of safety and efficacy is published . To thisThis meta-analysis was conducted in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA)  during tRelated articles published from inception to April 10, 2023 were selected, and PubMed, EMBASE, Cochrane’s Library and Web of Science were searched using the following terms: (1) “left bundle branch area pacing” and “biventricular” OR “cardiac resynchronization therapy” OR “CRT”; (2) “His-bundle pacing” and “biventricular” OR “cardiac resynchronization therapy” OR “CRT”; and (3) “conduction system pacing” and “biventricular” OR “cardiac resynchronization therapy” OR “CRT”. Details on search strategies are shown in Case report, abstract, review, editorial, letter or study involving less than ten patients were excluded. Inclusion criteria were as follows: (1) randomized or non-randomized controlled trial (RCT) and observational study; (2) studies comparing LBBaP or HBP with BVP among patients with HF; and (3) studies investigating one of the following outcomes: QRS duration (QRSd), echocardiographic parameters [left ventricular ejection fraction (LVEF) and/or left ventricular end-diastolic dimension (LVEDD)], NYHA functional classification, echocardiographic response, clinical and super clinical CRT response rate, pacing threshold, hospitalization rate of HF and all-cause mortality.Echocardiographic response was defined as an LVEF improvement of at least 5% at the end of the follow-up, and clinical response was defined as an improvement in the patient symptom, such as decreased NYHA functional classification of at least one grade at the last follow-up , while sTwo investigators independently screened the titles and abstracts, full text of potential eligible articles according to the inclusion criteria. Any disagreement was resolved by discussing with the third investigator.Two independent investigators conducted the data extraction from the included studies, and disagreements were resolved by consensus and discussions with the third investigator. The following information was extracted from studies: general study information , study characteristics , participant characteristics  and efficacy indexes .Two independent investigators assessed the quality of the included RCT with the Cochrane’s Risk of Bias Tool . Qualityσ2▲ = σ2pre + σ2post—2ρσpreσpost) was used [pre represents the variance at baseline; σpost, the variance at the end of the follow-up; and ρ, the correlation coefficient for correlations before and after the intervention. In accordance with practice, this analysis was conducted with an estimated ρ value of 0.5.Mean difference (MD) of the difference between the last follow-up and baseline was used to measure the outcomes of continuous variables, while odds ratio (OR) was used for the categorized variables. Once the variance for the paired difference before and after the intervention was not reported, the equation of  and R software (version 4.1.2) were used for the statistical analysis. A 2-sided Initially, a total of 126 potentially eligible articles were retrieved from the databases  had LBBB, while in 8 studies [n = 584). Meanwhile, 92% (n = 972) of the population used β-blockers in 11 studies [n = 847) used Angiotensin-Converting Enzyme Inhibitors (ACEI)/ Angiotensin Receptor Blockers (ARB)/ Angiotensin Receptor-Neprilysin Inhibitor (ARNI) in 12 studies [Ten of the included articles , 34–38 w studies , 38, the studies , 32–38,  studies –38. All  studies , 27–38 sP < 0.001, I2 = 91%). Similarly, in patients with LBBB, BVP presented a less reduction in a mean of QRSd than HPSP  in 8 studies [P < 0.001, I2 = 94%) . Consistently, LVEF in HPSP improved to a greater degree  than BVP in LBBB patients from 8 studies [P = 0.004, I2 = 35%). Among patients with LBBB from 5 studies [P < 0.001, I2 = 0%) . In patients with LBBB, HPSP also improved significantly compared with BVP  in 6 studies [Pooled analysis using a random-effect model from 9 studies , 37, 38  studies , 37, 38  studies , 37, 38  studies , 35, 37, studies , 32, 37 P = 0.01, I2 = 57%), while no significant difference was observed between HBP and BVP . Similarly, in LBBB patients, BVP had a greater change  than LBBaP in 5 studies [P = 0.43, I2 = 0%) in 3 studies [The difference in threshold change was assessed using the random-effect model in 6 studies , 37, 38  studies , 35, 37, studies , 32, 34 P < 0.001, I2 = 0%) echocardiographic response in 7 studies [P < 0.001, I2 = 0%) echocardiographic response than HPSP in 6 studies [P = 0.01, I2 = 0%) response than BVP, and there was no difference  between HPSP and BVP in LBBB patients from 5 studies [P < 0.001, I2 = 0%) response compared with BVP in 6 studies [P < 0.001, I2 = 0%) than HPSP in 5 studies [Pooled analysis was carried out using the random-effect model, and the results suggested that compared with BVP, HPSP patients were likely to achieve higher  [P < 0.001, I2 = 0%) [P = 0.09, I2 = 0%) [P = 0.31, I2 = 0%) [The hospitalization rate of HF was higher in BVP than HPSP in 10 studies  , 35–38 aI2 = 0%) , 38  , 35–38 aI2 = 0%) , 38 . Dense fibrous tissue surrounding the penetrating portion usually yields high capture threshold at implantation, and unpredictable rise in threshold may lead to premature battery depletion and lead revision. Meanwhile, LBB area is a thick band like structure that contains abundant left bundle branch (LBB) and Purkinje fibers, and electrical signals conduct across the LV rapidly. Therefore, LBBaP is provided with a unique advantage of low threshold. In the present research, LBBaP had a slower increase of threshold than HBP and BVP, and the battery of LBBaP could theoretically last longer . BesidesA total of 10 studies , 35–38 rIn conclusion, HPSP is associated with a shorter QRSd and thus a lower electromechanical dyssynchrony compared with BVP, and there may be fewer adverse cardiac events. Nevertheless, there are still some patients who are not suitable for class I recommendations in the guideline or do not response to current CRTs. In order to benefit from CRT, patients need to be screened and stratified more carefully. Additionally, more optimized therapies should be further explored for the treatment of the above patients.The studies included had good quality, and were from different countries with different clinical characteristics of the population. Another advantage of this analysis was that the differences between the baseline and the endpoint were adopted instead of only using the final value, which decreased the risk of confounding bias. The limitation of this study was that the proportion of RCT was not adequate. The heterogeneity might be explained by the small sample size, various follow-up duration and inconsistent baseline levels.In conclusion, the HPSP could produce shorter QRSd, higher LVEF and improved NYHA functional classification compared with BVP. Otherwise, LBBaP had a more stable threshold than BVP. Overall, these findings suggested the potential of HPSP as a promising frontline strategy for CRT. Further high quality RCTs should be performed to evaluate the efficacy between HPSP and BVP.Additional file 1.


Psychiatry is often perceived “different” by other medical professionals as well as by the general population. This perception of “difference” may give rise to stigma toward both patients with psychiatric disorders and mental health professionals.The aim of this study was to investigate the attitudes of non-psychiatric and their perception of psychiatry and mental disorders.It was a cross-sectional, descriptive and analytical study, conducted among Tunisian undergraduate and graduate non-psychiatric physicians. Data were collected during September and October 2022, through an anonymous online questionnaire, spread throughout social media (Facebook), using the Google Forms® platform. We used the “Attitude toward Psychiatry-30” (ATP-30).A total of 168 participants completed the questionnaire. Among them, 81  were undergraduate and 87  were graduate doctors. Their mean age was 26,4± 4.4 years, with a sex-ratio (F/M) of 3.4.Among doctors, 79,2% had overall favorable attitudes toward psychiatry but only 38,2% among the undergraduate considered psychiatry as a potential career choice.Psychiatry was considered as an unscientific and imprecise specialty by 20,3%; while 35,7% considered it as the least exciting.The total score ATP-30 increased significantly with age (p= 0.023). It was significantly higher in those with psychiatric history (p=0.01).Our study showed a dissonance between favorable perception of psychiatry and the choice of psychiatry as a potential career. Therefore, it is crucial to identify factors that potentially account for this dissonance and enhance enthusiasm among undergraduate doctors as the shortage of psychiatrists may influence mental healthcare.None Declared


Urological tumors, such as prostate cancer, renal cell carcinoma, and bladder cancer, have shown a significant rise in prevalence in recent years and account for a significant proportion of malignant tumors. It has been established that metastasis to distant organs caused by urological tumors is the main cause of death, although the mechanisms underlying metastasis have not been fully elucidated. The fibronectin receptor integrin α5β1 reportedly plays an important role in distant metastasis and is closely related to tumor development. It is widely thought to be an important cancer mediator by interacting with different ligands, mediating tumor adhesion, invasion, and migration, and leading to immune escape. In this paper, we expound on the relationship and regulatory mechanisms of integrin α5β1 in these three cancers. In addition, the clinical applications of integrin α5β1 in these cancers, especially against treatment resistance, are discussed. Last but not least, the possibility of integrin α5β1 as a potential target for treatment is examined, with new ideas for future research being proposed. Cancer remains one of the major contributors to death, with an estimated 19.3 million new cases and nearly 10 million cancer case deaths worldwide in 2021. Among the most prevalent cancers, prostate cancer ranks first, bladder cancer fourth, and renal cell cancer sixth in incidence rates among men . During in vivo, mediating cell binding to the extracellular matrix and producing signals associated with various diseases, such as autoimmune responses, tissue and organ development, cardiovascular disease, and cancer  completely inhibited the growth and cell morphological alterations of prostate cancer PC-3 cells, confirming that integrins interact with the FNIII10 structural domain and play a key role in these processes  is a single-chain polypeptide that mediates endocrine, autocrine, and paracrine growth, thus acting as a potent growth factor. IGF-1 acts on cells by binding to its receptor, IGF-IR, a transmembrane protein with tyrosine kinase activity . The insr growth . This phr growth .Adhesion to fibronectin and its fragments, migration, and invasion of prostate cancer cells via integrin α5 is considered one of the mechanisms by which bone marrow localization is regulated by bone-derived mesenchymal stromal cells , 75, 76.Cytoskeletal organization and adhesion formation are essential for cell motility and structural support . IntegriLOX-like protein 2 (LOXL2) is a member of the lysyl oxidase family, consisting of Lox and four Lox-like proteins, with intracellular and extracellular functions. The secreted LOXL2 regulates integrin levels to promote tumor progression in renal clear cell carcinoma (ccRCC) by regulating lysyl oxidase LOXL2 status and its correlation with tumor staging . It was MicroRNAs are endogenous non-coding RNAs found in eukaryotes that have regulatory functions. miRNAs consist of 21–25 nucleotides, are extensively involved in the pathogenesis of human cancers, and can function as oncogenes and tumor suppressors . IntegriDuring clinical practice, drug resistance is commonly observed in the treatment of tumors, and studies have shown that integrin α5β1 is involved in this phenomenon. Juengel et al. found that integrin α5 was involved in the development of renal clear cell carcinoma resistant to mTOR inhibitors, characterized by quantitative changes in integrin α5 expression during drug resistance and coupled with altered molecular function of integrins, forcing renal clear cell carcinoma to shift from adhesion to migration . SimilarIntegrin α5β1 represents a promising therapeutic target for tumor angiogenesis and tumor cell expression. Although most studies involving α5β1 inhibitors are in the preclinical or pre-phase III clinical trial stages, many specific inhibitors have been successfully developed. These inhibitors hold huge potential for use against specific subgroups of aggressive tumors at this stage. Given that integrin α5β1 can enhance angiogenesis, the main function of these related inhibitors is anti-angiogenesis, with the development of specific antibodies and small peptides that target this pathway Table 2.in vitro and inhibiting prostate cancer growth, metastasis, and tumor recurrence. In phase I clinical trials, systemic PHSCN peptide monotherapy demonstrated good tolerability, with metastatic disease progression delayed by 4–14 months in one-third of the treated patients (P < 0.02), with significantly fewer hepatic tumor microvessels in the ATN-161 and ATN-161, 5-fluorouracil groups than in the control and 5-fluorouracil groups (P < 0.05). The combination of ATN-161 and 5-fluorouracil was more effective than either treatment alone, with a significantly increased apoptosis rate of tumor cells, inhibited proliferation of tumor cells (P < 0.03), and improved overall survival rate (P < 0.03) (ATN-161 (Ac-Pro-Her-Ser-Cys-Asn-NH2) is a small pentapeptide-containing cysteine that mainly binds to the arginine–glycine–aspartate (RGD) structural domain and acts as an inhibitor of integrin α5 . After tpatients . In anot < 0.03) . OverallVolociximab is a chimeric human α5β1 antibody that consists of a variable region of mouse antibodies, including a complementarity-determining region against the α5β1 integrin, combined with a constant region of human IgG4 heavy chain and kappa light chain. In preclinical models, volociximab has shown the ability to prevent neovascularization by inhibiting fibronectin binding, has similar affinity and activity to integrin α5, and has been shown to be safe, effective, and tolerable . MoreoveJoshi et al. prepared and analyzed a bispecific antibody (BsAbα5β1/αv) in prostate cancer that targets the degradation of both α5 and αv integrins. The results showed that this antibody was superior to monoclonal antibodies in eliminating prostate cancer cell adhesion, migration, and clonal survival .SJ749 is a non-peptide inhibitor of integrin α5 that inhibits angiogenesis by affecting endothelial cell adhesion and migration . It has JSM6427 is also a potent, highly specific inhibitor of integrin α5 . This drin vitro (Many blocking antibodies have been developed against the interaction between integrin α5 and fibronectin—for example, IIA1, a functionally blocking mouse antibody to integrin α5, can inhibit angiogenesis, cell adhesion, invasion, and tumor cell survival in vitro . MINT152in vitro  (Table Research on integrin α5β1 inhibitors in recent years has helped to demonstrate the specificity of integrins and the ability of these inhibitors to control multiple oncogenic pathways. The development of integrin inhibitor compounds is essential for developing new and more effective therapeutic options for treating urologic tumors. Most integrin α5β1 inhibitors are currently in phase 1 and phase 2 clinical studies, and it is essential to initiate phase 3 clinical trials as soon as possible . There aThis review describes the structure, function, and association of integrin α5β1 with prostate cancer, renal cell carcinoma, and bladder cancer as well as the clinical applications and research in these diseases. Integrin α5β1 is closely associated with cancer development, progression, and prognosis. Dysregulation of integrin α5β1 is strongly related to the development of urological tumors and may serve as an important indicator for evaluating invasion and migration. Additionally, the upregulation of integrin α5β1 can promote the development of drug resistance in cancer cells, which prompted the exploration of novel strategies for overcoming drug resistance in chemotherapy. The overexpression of integrin α5β1 has been documented in various tumors and reportedly contributes to tumor progression, making it a potential target for tumor imaging and an independent indicator of poor prognosis. Besides this, it has been extensively explored as a tumor suppressor in preclinical or clinical studies. Nonetheless, integrin α5β1 has not yet been established as a definitive molecular biomarker that can be applied to specific diseases, representing an area for future research. Therefore, research on integrin α5β1 can help to improve the diagnosis and treatment of this patient population.GuZ conceived the manuscript. XuZ searched publications and drafted the manuscript. HZ, HX, and CL edited the tables and figures. XiZ and JZ reviewed the manuscript and polished the grammar. All authors contributed to the article and approved the submitted version.