Polymers constitute a group of materials having a wide-ranging impact on modern pharmaceutical technology. Polymeric components provide the foundation for the advancement of novel drug delivery platforms, inter alia orodispersible films. Orodispersible films are thin, polymeric scraps intended to dissolve quickly when put on the tongue, allowing them to be easily swallowed without the necessity of drinking water, thus eliminating the risk of choking, which is of great importance in the case of pediatric and geriatric patients. Polymers are essential excipients in designing orodispersible films, as they constitute the backbone of these drug dosage form. The type of polymer is of significant importance in obtaining the formulation of the desired quality. The polymers employed to produce orodispersible films must meet particular requirements due to their oral administration and have to provide adequate surface texture, film thickness, mechanical attributes, tensile and folding strength as well as relevant disintegration time and drug release to obtain the final product characterized by optimal pharmaceutical features. A variety of natural and synthetic polymers currently utilized in manufacturing of orodispersible films might be used alone or in a blend. The goal of the present manuscript was to present a review about polymers utilized in designing oral-dissolving films. Among the many different routes of drug application (the Food and Drug Administration (FDA) distinguishes about 100 of them), the most preferred and the least invasive is the oral route ,3,4,5,6.2 [Orodispersible films (ODFs) represent an innovative system of oral administration of medicines in the form of a thin, polymeric, oral matrix having a thickness in the range from 12 to 100 µm and surface of 2 to 8 cm2 ,6,7,9,112 . Oral fi2 ,9,14,15.w/w) affected the ropiness of ODFs. In vivo organoleptic investigations showed that films made of hypromellose (Pharmacoat 606) and plasticized with polyethylene glycol or a mixture of glycerol and polyethylene glycol were of unsavory taste, while ODFs made of glycerol alone had a more favorable taste than films with propylene glycol. Glycerol ensures improved plasticity of films formed utilizing polyvinyl alcohol, while diethylene glycol might be utilized for ODFs composed of either HPMC or PVA [Plasticizers are an important group of excipients used in the development of ODFs, usually adding up to 20% by weight of the dry polymeric powder. Examples of plasticizers are as follows: glycerol, propylene glycol, mannitol, sorbitol, diethyl phthalate, tributyl and triethyl citrate, macrogol, or esters of citric acid ,14,15,16C or PVA .Examples of surfactants that improve spreading of the casted solution and wetting by saliva in the mouth, utilized in manufacturing of ODFs, are polysorbate, poloxamer 407, sodium lauryl sulfate, and examples of sweetening/taste-masking agents are flavored essences, sorbitol, mannitol, glucose, aspartame, fructose, sucralose, cyclamate, sucrose, maltose, oleoresins, ribose, thaumatin I and II. As dryness in the oral cavity delays the rapid dissolution of ODFs, it is vital to add salivatory secretion stimulants in order to increase the rate of saliva production. For this purpose, mainly organic acids are used: malic, ascorbic, tartaric, or lactic acid ,9,15,16.Polymers are considered as excipients, but they have grown to be a key element in designing of the films, being responsible for a number of their properties. Thus, comprehension of polymers’ physicochemical characteristics and rheology is essential to optimize their utility in ODFs’ preparation. It is necessary to consider a choice of the suitable ones to produce the film, ensuring the most advantageous therapeutic result ,9,15,16;The viscosity of the polymer solution is crucial for ODFs’ production and drug release from the designed formulation. The polymeric materials that create a thick gel layer of high viscosity upon wetting will result in slower release compared to polymers of low or intermediate viscosity. On the other hand, solutions having an insufficient viscosity are found to be difficult to cast and obtained films are difficult to handle due to air bubble entrapment. Various viscosity limits are reported in the literature, but these cannot be used as overall recommendations as the required viscosity level is highly dependent on the manufacturing method and equipment used. It is essential to keep in mind that viscosity depends not merely on the quantity of polymer used, but also on its type; therefore, the individual limits should be assessed for each polymer or combination of polymers. A considerable difficulty in the designing of ODFs is to predetermine the optimum viscosity range of the casting compounds. Furthermore, their viscosity, however highly influential, is not the single parameter that affects the behavior of the mass in ODFs’ development, as other factors, associated with the structure of the polymer chains, the interactions of the ingredients, the nature of the solvent or the surface tension, are also influential. Further variations in the characteristics of the casting mass are caused by the presence of the drug. In a suspension-type system formulated by insoluble drugs, further difficulties occur in relation to adequate viscosity adjustment so that sedimentation of particles in the film-casting process does not proceed too rapidly. The viscosity of the casting solution also needs to be chosen depending on the height of the casting gap, with the principle that the thicker the layer of solution applied, the higher viscosity is demanded ,19,20.Primarily, water-soluble polymers  are employed in formulation of ODFs to obtain a thin matrix characterized by rapid disintegration time, good mechanical properties, and organoleptic sensations. Commonly utilized ones include hydrophilic cellulose derivatives (hydroxypropyl methylcellulose (HPMC), hydroxypropyl cellulose (HPC), polyvinyl alcohol (PVA), polyvinylpyrrolidone (PVP), or polyethylene oxide (PEO)). Recently, the polymers of natural origin, chiefly polysaccharides, have received significant interest in the pharmaceutical industry on account of being safe, biodegradable, and biocompatible. In this regard, either maltodextrin, chitosan, or pullulan is employed. Hydrophilic materials are the leading preference for preparing the ODFs, so that the drug dosage form can dissolve easily and softly in the mouth. Hydrophobic polymers such as methacrylic acid copolymers or polyvinyl acetate (PVAc) are also used, although less frequently ,18,19,206H10O5)n, consisting of linear chains formed by β-D-glucose units connected by β-1,4-glycosidic bonds CH3 is a hydrophilic group devoted to the hydration rate, while the methoxy group is considerably more hydrophobic. This feature is considered to be essential for dissolution of drugs and their release profile from ODF. In general, HPMC grades with high viscosity and methoxyl content contribute to obtain more robust, rigid, and stretchable films. Higher viscosity is connected with greater branching and/or higher MW related to physical entanglement due to longer chains. This effect might raise the input strength necessary to break the native chain interactions, resulting in higher tensile strength of the film. In contrast, higher degrees of methoxyl substitution may result in an entrenching effect on HPMC chains due to their larger size relative to the primary hydroxyl groups, which may also lead to high tensile strength. However, it should be emphasized that the degree of methoxyl substitution does not considerably influence the Young’s modulus . Regarding the thermal properties of HPMC films, the polarity of the polymer chains resulting from the methoxyl content impacts the Tg. HPMC varieties with less methoxyl groups are characterized by lower polarity, which reduces the free area between polymer chains. The growing distance between the polymer chains results in intensified secondary interactions between the chains, which enhances the energy needed to translocate the chains [HPMC consists of partially O-methylated and O-(2-hydroxypropylated) cellulose, and is accessible in a number of types, differing in molecular weight and number of substituent groups in the anhydroglucose units. The mean amount of methoxy and hydroxypropoxy groups bonded to the ring is expressed as the DS and significantly affects the properties of the oral film, especially in relation to the release of a drug and mechanical features. The hydroxypropoxy group —OCHe chains ,19,23.3) and the latter two relate to the approximate percentage of a more hydroxypropoxy group  and USP define various types of HPMC substituents using a four-digit number, HPMC 1828, HPMC 2208, HPMC 2906, and HPMC 2910, in which the initial two digits relate to the approximate percentage of a more methoxy group  anw/w) at 20 °C and marked as C (multiplier of 100) or M (multiplier of 1000). The last suffix is needed to verify the grade of product, such as premium (P), low viscosity (LV), controlled release (CR), granular (G), direct compression (DC), surface treated (S), or food grade (FG). The polymeric material is traded under various brand names, for instance, Metolose and Pharmacoat , Methocel , and Benecel  [Such variations in the ratios of methoxy and hydroxypropoxy substituents and molecular weight influence their features such as organic thermal gelation temperature in aqueous solution, swelling, solubility, drug release rate, and diffusion. Practically, basic hypromelloses are labelled as type 2208—K, 2910—E. A suffix is additionally given to designate the viscosity of the polymer solution, which corresponds to the molecular weight of the HPMC. It is measured in a 2% aqueous solution (erlands) ,24,25,263. Pure HPMC film features a smooth, homogeneous, and uniform matrix. The structure of the HPMC film is greatly affected by the quality of the bonds in the film matrix, which are carried out throughout the drying process [Basically, type E rather than K is utilized in preparation of the ODFs. Especially, grades E3, E5, and E15 are widely utilized, particularly with regard to their low viscosity and optimal Tg, respectively 160 °C, 170 °C, and 175 °C. The major distinction among these grades is the length of the polymer chain, which is related to the increase in MW of HPMC (E3 < E5 < E15 < E50). The maximum puncture strength increases with MW E3 < E5 < E15 < E50. However, it is stated that E3 and E5 may result in thin, brittle, and non-peelable films. Such features might be improved with increasing concentration of the polymers, but the films are still described as tacky; hence, a blend of various HPMC grades is favored, especially those with higher MW. For instance, E15 has beneficial film-former properties . However, HPMC–maltodextrin mixtures, with higher maltodextrin content, produce thin, fast disintegrating, and palatable films, characterized by an appropriate degree of sweetness. The temperature of degradation of HPMC is in the range from 200 °C to 250 °C (depending on side substitutions of the polymer chain). Considering these characteristics, including high Tg and low temperature of polymer degradation, it should be highlighted that most of the available HPMC grades are not stable during heat treatment ,23,27. H process ,30,31.w/w), compositions containing AP05:AP40:PVP in a ratio of 5:2.5:7.5 or 4:3.5:7.5 were not viscous enough. The viscosities of the HPMC and PVA/PVP solutions chosen for subsequent investigation were 1.9 and 0.76 Pa∙s, respectively. However, such solutions did not enable the preparation of placebo films (deprived of a drug) with uniform thickness and smooth surface from a 1000-µm gap, but using a smaller one (800 µm and less) resulted in proper film formation. On the other hand, the higher viscosity of the solutions was disadvantageous in terms of a venting problem and less uniform mass distribution on the plate [w/v) was chosen to prepare everolimus ODFs plasticized with propylene glycol, PEG400, or glycerol [w/w)-based ODFs’ films containing prednisolone encapsulated in spray-dried microparticles made of Eudragit E PO with glycerol or PEG 200 as plasticizer. Films containing glycerol at 3.125% or 3.75% concentration were brittle and fragile, while formulations containing PEG 200 were found to be more flexible and resistant. However, ODFs containing PEG 3.75% were too viscous and ductile, so a formulation of 12.5% HPMC and 3.125% PEG was selected as optimal [w/v) were used as the film-forming agents to fabricate phenytoin-loaded ODFs and they were evaluated in terms of extrudability and printability by a syringe extrusion by 3D printing method. Glycerin and propylene glycol were utilized as plasticizers. The diameters of the printed fibers were found to decrease with increasing concentrations of HPMC E5 and HPMC E15. However, it was also observed that the inclusion of the drug affected the viscosity and printability of the prepared dispersion. The formulations prepared from HPMC E15  with phenytoin were not extruded smoothly through the nozzle because of its high viscosity and fast drying of the printing dispersion, while the formulations of HPMC E5  and type E15  were effectively extruded through the nozzle. The consistency factor scores of HPMC E5 were found to be lower compared to those made of HPMC E15 at the same concentration, suggesting that HPMC E15 shows non-Newtonian pseudoplastic behavior as well as optimal extrudability properties through the extrusion nozzle. The viscosities of 10, 12.5, 15, and 20% (w/v) of HPMC E5 were 0.80 ± 0.04, 1.70 ± 0.03, 3.02 ± 0.18, and 9.84 ± 0.08 Pas and the viscosities of 10 and 12.5% (w/v) of HPMC E15 were 8.10 ± 0.51 and 16.85 ± 0.76 Pas, respectively. It was evaluated that higher viscosity value was observed when the drug was incorporated into the formulations. In the case of E5 20% and E15 10% formulations, after phenytoin was incorporated, the viscosities were determined to be 13.39 ± 0.74 and 12.17 ± 0.47 Pas, respectively. Furthermore, the findings demonstrated that, for a lower-molecular-weight polymer (HPMC E5), the higher polymer concentration is necessary to obtain a similar viscosity as the high-molecular-weight polymer (HPMC E15). Then, printed ODFs characterized by optimal mechanical properties and the fastest disintegration time were selected to evaluate their drug content and dissolution profiles. The results showed that ODFs based on HPMC E15 demonstrated superior properties when compared to E5 films. Fast disintegration time in less than 5 s and rapid dissolution (up to 80% of drug was released within 10 min) were demonstrated. In addition, a formulation made of HPMC E15 exhibited drug content uniformity, good mechanical properties, flexibility with low puncture strength, low Young’s modulus, and high elongation, which enables easy handling and application [The literature provides a number of experiments that led to the creation of films based on HPMC. The following polymers were compared in the research to obtain ODFs containing loratadine hydrochloride by solvent-casting method: HPMC (Pharmacoat 606) in concentration of 15% and combination of PVA (7.5%) and PVP (7.5%). All formulations were plasticized with 3% polyoxyethylen glycol (PEG). It has been established that 15% solution of HPMC and a 15% solution of a mixture of PVA05, PVA40, and PVP in a ratio of 2.5:5.0:7.5 are optimal for incorporation of the drug. In the initial trials it was determined that films with high PVP content tended to be too brittle and a higher PVA ratio resulted in both too brittle and too flexible formulations. The mixture of PVP and PVA in the selected proportion led to an appropriate flexibility of the films supplied by PVA and a reduction in disintegration time because of the content of PVP. It was also significant that at the tested polymer concentration in the casting solution, i.e., 15%  polymer concentration, had the following composition: F1, HPMC E3; F2, HPMC E5; and F3, HPMC E5 (26%) along with NaCMC (4%). It was assessed that all ODFs dissolved quickly and were readily disintegrated, owing to the hydrophilic nature of polymeric materials used. However, changes in the drug release profiles were noted, which were probably associated with variations in the polymer type: A significant increase was observed for the F3 formulation, which released 93% of the drug under 8 min. The formulation F3 dispersed quickly in the medium and homogenic distribution of drug particles was observed. An addition of NaCMC greatly influenced the percentage of released dose, as the combination of HPMC and NaCMC increased the hydrophilic characteristics of ODFs. Additionally, the formulation F3 exhibited high roughness of the surface and porosity, which might be related to rapid drug-release behavior. Hydrogen-bonding interaction between drug and polymer were revealed by spectral analysis [HPMC of different grades  and NaCMC, all plasticized with PEG 600 and glycerol, were evaluated for the ability of creating ODFs with donepezil. The formulations (F1–F3), with 30%  in different combinations plasticized with glycerol 85%, were used to prepare bilayered ODFs containing enalapril maleate . It was Explored ODFs’ compositions based on cellulose derivatives are shown in Within all naturally occurring biopolymers, starch has been regarded as one of the most prospective polymers due to its widespread availability, biodegradability, and relatively low cost. It consists of two polymers, namely, amylose  and amylopectin  . A varieStarch granules consist of three particular regions, which are the amorphous region, the crystalline lamellae, and the amorphous growth ring. Considering the stable semi-crystalline framework of starch, the granules are not dissolved in water at room temperature. Nevertheless, it is subjected to restricted swelling, probably as a result of hydration and swelling of the amorphous regions. It is recognized that amylopectin, because of its highly branched and open structure, enables hydrogen-bonded solvent molecules to access its composition. Therefore, in water or any other solvent capable of forming hydrogen bonds, it undergoes more disruption than amylose, which features a highly packed structure. The interaction of starch granules with hot water is distinct from that of room temperature. Exemplary, when starch granules are heated in excess water above a critical temperature that depends on the type of starch, an irreversible change called gelatinization occurs. Applicability of starch is restricted due to its efficient barrier against low polarity compound and its poor mechanical strength. Plasticizers are generally required for oral films composed of starch to overcome film brittleness. Usually, employed plasticizers for starch films are glycerol and sorbitol. In general, low concentrations and high temperatures are needed to dissolve the starch. Process-related problems also concern the difficulties connected with solubility of native starch in water, owing to its large molecular size and strong hydrogen bonds. Therefore, to surmount this drawback and to enhance the performance of the ODFs, various starch derivatives have been engineered. Exemplary modified starches used in the formulating of ODFs include hydrolyzed starches such as pregelatinized starches, maltodextrins, or pullulan ,51,52.Pregelatinized starch is chemically and/or mechanically modified starch, commercially available in fully or partially pregelatinized form. The first one is easily soluble in cold water, while the second one has soluble (gelatinized) but also insoluble fractions. Pregelatinized starch features good film-forming properties, creating transparent, robust, and elastic films using a solution at a concentration of 15% to 20% in the solvent-casting process ,51. Lycow/w) created products with favorable peelability, whereas at the lower concentration of 10% (w/w), no film creation was noted. PG and polysorbate 80 were used as plasticizers. PG in concentration 2% (w/w) gave fragile ODFs with low folding strength; therefore, its concentration was raised to 4% (w/w), resulting in elastic matrices with optimal folding strength. In both trials, the concentration of polysorbate 80 was constant: 2% (w/w). Nevertheless, such composition did not allow the obtained films to disintegrate, so PG was substituted with sorbitol , which shortened the disintegration time. It was later noted that when combining with soya lecithin, polysorbate was needed at a lower concentration. Therefore, 0.1% (w/w) polysorbate 80 and 0.8% (w/w) soya lecithin were added to promote wetting and disintegration (which occurred within 30 s). Additionally, obtained films were smooth, transparent, and non-tacky, having a thickness of 0.2 mm. Pullulan created a well-peeled film, both at concentrations of 2% (w/w) and 5% (w/w) with an in vitro disintegration time of less than 30 s; hence, the lower pullulan concentration was selected for further studies. Sorbitol was chosen as a plasticizer, as it provided a satisfying folding endurance. Drug-loaded pullulan films were non-tacky, transparent, and disintegrated within 15 s. The 7% (w/w) HPMC E-5 with PG 1% (w/w) formed well-peelable, fast-disintegrating (10 s) films. In the case of Kollicaot IR, a concentration of 10% (w/w) was chosen, as ODFs made of 5% (w/w) softened and at 2% (w/w) the matrices were not formed. There was no need to plasticize the films, as Kollicoat IR is composed of 75% polyvinyl alcohol and 25% of PG; therefore, only a wetting agent, polysorbate (0.1%), was applied. Obtained ODFs were non-tacky, flexible, transparent, and disintegrated within 21 s [An interesting comparative study, including multiple polymers, was carried out by Kathpalia et. al. . The goahin 21 s .Maltodextrin is an unsweet, water-soluble polysaccharide composed of D-glucose units generally associated with an α (1→4) glycoside bond, combined in chains of variable length, with a dextrose equivalent (DE) below 20, produced by the partial hydrolysis of native starch . It is eThe unique feature about the films crafted from maltodextrins is that they are extremely thin and elegant. In addition, the use of maltodextrin together with microcrystalline cellulose leads to the formation of non-sticky films with a smooth surface ,18,51. MAureobasidium pullulans. It is a modified starch consisting of glucose units in maltotriose linked by α(14) glycosidic bonds, whereas further maltotriose units are linked by α(16) glycosidic bonds . For membranes containing HPMC, a greater fiber diameter and a uniform diameter distribution were noted in comparison with pectin and NaCas membranes. The highest viscosity values obtained for HPMC membranes could be ascribed to the existence of a hydroxyl group in their molecule, which enhances the water-binding capacity, resulting in fibers with larger diameters. The incorporation of HPMC and pectin led to the quick disintegration of the membranes in 5 s, while the other formulations were over 15 s. However, the membranes made of a pullulan/HPMC mixture were considered as the best candidate for ODF formulations due to their rapid disintegration in a simulated saliva environment in less than 5 s, allowing convenient administration, as well as the complete release of the drug within 30 s, making the drug easily absorbed in the mouth and throughout the gastrointestinal tract after being swallowed [Transparent, elastic, and smooth ODFs of great solubility and optimal mechanical strength are obtained from pullulan. An important feature is that pullulan films’ flexibility is accompanied by high tensile strength and stability over a broad temperature spectrum. However, as the substance is sourced from the enzymatic fermentation process of yeast, its low availability (due to complex acquisition) makes it a highly expensive material. Hence, it is typically mixed with alternative polymers that are freely available and less expensive. In general, 50% to 80% of pullulan may be substituted with starch or modified starch without losing its desired performance as a favorable film-former excipient. Sodium alginate or carboxymethylcellulose (CMC) might be employed as well, due to their biocompatibility. Chiefly, the formation of hydrogen bonds between -COO alginate and CMC groups with –OH of pullulan can synergistically improve the material properties of the resulting films. Moreover, pullulan–HPMC blends, with HPMC content above 50%, were also reported and the final polymeric matrix showed better thermal and mechanical properties. The mechanical behavior of pullulan matrices is influenced by the temperature at which it was produced. ODFs formulated at low temperatures are stiffer and more flexible than those made at higher temperatures, which are brittle and do not exhibit pronounced plastic deformation. ODFs made of pullulan are also characterized by a fast disintegration time ,51,60,61wallowed .Sodium alginate and chitosan  are usedAbelmoschus esculentus L., a plant extensively farmed in tropical, subtropical, and temperate regions throughout the world. Okra extracts contain acidic polysaccharides consisting of various sugars including galactose, rhamnose, galacturonic acid, galactose, glucose, and glucuronic acid. Moringa gum is obtained from Moringa oleifera . It is a polyuronide composed of arabinose, galactose, and glucuronic acid in the ratio of 10:7:2, with rhamnose present in trace amounts. Okra and moringa gums have been tested for film-forming properties after blending with pullulan or HPMC to prepare ODFs with citalopram. It was noted that only okra biopolymer might be used in combination with HPMC or pullulan for producing appropriate formulations. Obtained ODFs were smooth, apart from those produced with HPMC and moringa gum combination, which were slightly rough in texture. The disintegration time was less than 30 s and the homogeneity of the drug content was 97.89–102.05% for all formulations. Additionally, the films made of okra gum and HPMC were characterized by superior mechanical properties compared to okra and pullulan blend or moringa gum and HPMC mixture [An interesting example of novel biopolymers are okra and moringa gum. Natural gums are chemically inert, environmentally friendly, non-toxic, biodegradable, biocompatible, and non-irritating. They also have the advantage of being easy to access and they are cheaper than synthetically modified gum. Okra is typically obtained from  mixture ,69.The polymer is non-toxic (authorized by the FDA for human consumption and considered as GRAS) with high water solubility . The matw/w) with variable content of glycerol by solvent casting technique. PVA (15%) with 5% glycerol content showedthe most relevant results. Obtained films were smooth with uniform surface morphology, possessed good tensile strength and disintegration time (ranged from 7.7 to 28 s) and released more than 50% of drug within first 2 min [w/w) to prepare ODF with phenytoin by solvent casting method. From the obtained formulations, these consisting of 1% (w/w) PVA, 0.04% (w/w) sodium starch glycolate and PEG 400, glycerin and water as cosolvents led to development of promising products. The physical appearance (transparent and colorless ODF with a smooth surface), mechanical strength, drug content and disintegration time were found to be optimal [PVA-based ODFs plasticized with PEG and glycerin containing salbutamol sulphate were obtained by solvent casting technique. ODFs were transparent with smooth surface, characterized by optimal mechanical properties and demonstrated an immediate release of 90% within the first 2 min. The dissolution profile and physico-mechanical features such as appearance, surface of pH, softness and elasticity were accessed as good . PVA as st 2 min . The aim optimal .Polyvinyl pyrrolidone is a non-ionic, synthetic polymer composed of linear 1-vinyl-2-pyrrolidinone groups  obtainedODFs with PVP as a polymeric backbone were created with enrofloxacin by solvent-casting method, exhibiting good uniformity and mechanical properties. Their surface pH was 6.8, indicating that these formulations would not cause irritation to the oral mucosa. Average time for the film to disintegrate into completely fine particles was about 1.26 min .Polyethylene oxide is an example of further synthetically derived polymeric material , which f® VA 64) and poloxamer types 407 and 188 on 3D printer. All of the obtained ODFs were characterized by a uniform drug content, optimal strength, and rapid disintegration time and exhibited good printability. The formulation containing PEO, PVP, and poloxamer 188 indicated the fastest dissolution time and immediate dissolution profiles [Olanzapine ODFs were developed from PEO as film-forming polymer with the addition of PVP (Kollidonprofiles .w/w) of film weight, was achieved [Most commercially available and laboratory-created oral films are usually made of hydrophilic film-forming polymers. By definition, hydrophilic components possess a greater affinity for water, in comparison to hydrophobic substances. In this regard, the physical, chemical, and microbiological stability of hydrophilic matrices can be severely affected by moisture if they are not protected by appropriate packaging. When directly exposed to regular environmental humidity, these hydrophilic structures can become sticky when the relative humidity is high or brittle when the relative humidity is very low. In either case, it leads to adverse texture and appearance of the obtained product. At the molecular level, small or less perfect crystalline polymer structures can be lost, contributing to lower Tg. In contrast, hydrophobic polymers are known to absorb less water even at high moisture content at room temperature. The hydrophobic nature appears to be incompatible with rapid disintegration in the case of ODFs, so the presence of a channeling agent  by promoting fluid penetration can provide rapid disintegration ,78. Howeachieved . An examachieved .ODFs continue to be a current concern for technologists and researchers, who concentrate on their continuous development. The pharmaceutical market is increasingly abundant with the availability of a variety of ODFs. The instances of commercially available ODFs and excipients utilized in their manufacturing are given in Polymers are regarded as essential excipients in the pharmaceutical technology, having a significant influence on the advancement of ODFs. Concerning the properties of the polymeric material, ODFs disintegrate or dissolve spontaneously in the mouth, which results in the formation of a solution, soft paste, or suspension without the necessity for fluids’ intake. When designing ODFs, attention should be paid to the proper selection of polymers. The overall purpose is to implement cost-efficient, biocompatible, versatile, non-toxic polymers. Among different types of polymers, their blends are usually used so that they can mutually complement their properties, as polymeric materials of lower molecular weight disintegrate faster, while using higher molecular weight polymers provides ODFs with improved mechanical features. In this regard, it is essential to ensure the right balance between the mechanical characteristics and the disintegration time demanded for ODFs. The multitude of polymeric materials’ types, different polymer classes, and various polymer-to-polymer ratios result in an explosive amount of different formulations and a broad spectrum of ODFs’ specifications. Consequently, it is extremely important to have a thorough comprehension of the system under development to prevent obtaining dosage forms with undesirable properties. This manuscript summarizes the role of polymers alone or in combination and their impact on the designed formulation and provides insight into their functionality in the process of ODFs’ manufacturing.


Burundi has experienced an increase in malaria cases since 2000, reaching 843,000 cases per million inhabitants in 2019, a more than twofold increase compared to the early 2000s. Burundi thus contrasts the decreasing number of cases in many other African countries. To evaluate the impact of malaria control on this increase, data on interventions from 2000 to 2019 were compiled. Over this period, the number of health facilities increased threefold, and the number of tests 20-fold. The test positivity rate remained stable at around 50–60% in most years. Artemisinin-based combination therapy was introduced in 2003, initially using artesunate–amodiaquine and changed to artemether–lumefantrine in 2019/2020. Mass distribution campaigns of insecticide-treated bed nets were conducted, and indoor residual spraying and intermittent preventive treatment in pregnancy introduced. Thus, the increase in cases was not the result of faltering control activities. Increased testing was likely a key contributor to higher case numbers. Despite the increase in testing, the test positivity rate remined high, indicating that current case numbers might still underestimate the true burden. In 1990–1991, prior to any major interventions, mortality due to malaria in infants below 1 year was 38 per 1000, and 8 per 1000 in children aged 1–4 years [Burundi is a small 27,834 km7,834 km2Since 2000, a pronounced increase in cases has been observed, resulting in epidemic transmission starting in 2017. Over 9 million cases among the population of 11.4 million were recorded in 2019 . The patData on health care and services are collected daily at the community level, in health centres, and hospitals. Every functional health facility in Burundi has the duty to transmit data on the standard national health information systems (HIS) tools . Until 2Possible poor quality and lack of completeness of data is a limitation of this report and hampers surveillance in Burundi. According to the National Health Information System Data Quality Improvement Strategic Plan 2019–2023, in 2018, the overall completeness of health facility reports was 92%. Only one health district completed < 80% of reports. None of the monthly reports from the health zones had zero/missing values . Over thP. falciparum cases increased threefold from 3,272,608 to 9,416,989. Thus, the cases per million population increased from 513,000 to 843,000. The number of confirmed deaths per year per million increased from 60 to 110 in 2000–2004 to 300–600 in more recent years  were introduced, and by 2018 were used for over 75% of malaria diagnosis. Across all years, test positivity rates by RDT and microscopy were nearly identical , suggesting similar diagnostic sensitivity of the two methods. Different brands of RDT were used in Burundi, as funders made their own choices under the condition that the RDT is prequalified by the WHO. All health providers at public and private health centres are trained in RDT and microscopy use. The national malaria control program organizes quarterly formative supervision on laboratory test quality assurance targeting the private and public sector.From 2003 to 2019, artesunate-amodiaquine (ASAQ) was used as first-line treatment. Artemisinin therapeutic efficacy trials have been conducted in Burundi three times. In 2002 the treatment response rate for ASAQ and artemether–lumefantrine (AL) was > 95% and > 99% respectively . ArtesunDistribution of long-lasting insecticidal nets (LLINs) started in 2003 through social marketing campaigns. Since 2005, distribution of LLINs were integrated with other routine health centre-based services . LLINs mass distributions campaigns in Burundi were initiated for the first time in 2009, targeting the most vulnerable people (children under 5 and pregnant women). Distribution was integrated to measles vaccination campaigns; 1.6 million LLINs were distributed among the 8.4 million inhabitants. Further mass distribution campaigns took place in 2011, 2014, 2017, and 2019 . Deltamethrin-based LLINs (Yorkool®) were distributed in all the remaining districts.All LLINs distributed until 2018 were permethrin-based LLINs (Olyset Net®), alphacypermethrin-based LLINs , and deltamethrin-based LLINs . In 2019, pyrethroid-PBO LLINs were distributed in 4 health districts with identified The LLIN coverage after the 2014 mass-distribution campaign was estimated at 97%, and after the December 2019 campaign at 96.7%. However, the durability monitoring early findings demonstrated that the LLINs survivorship in the households was 63% for the standard LLINs and 88% for the PBO LLINs 8 months after the mass-distribution campaign [The campaigns in 2011 and 2017 might have resulted in a reduction in case numbers. From 2011 to 2012, the number of tests increased from 340,000 to 439,000 per million, yet case numbers decreased from 380,000 to 290,000 per million. Test positivity rate in 2012 was the lowest over the period studied at 34.5%. From 2017 to 2018, the number of tests decreased from 810,000 to 684,000 per million. Case numbers declined from 759,000 to 522,000 per million, resulting in a moderate decrease of the test positivity rate from 63 to 51%. After the 2014 campaign, the number of tests decreased, but the number of cases increased moderately . According to the UNHCR, from 2015 to 2017, over 420,000 Burundians have left the country, and a further 55,000 were displaced within the country . FurtherResistance has been documented against all available antimalarial drugs, and is a constant threat to malaria control. Emergence of indigenous resistance to the first-line drug AL has been documented in Equatorial Guinea , and muthrp2 deletions, resulting in false-negative tests. Deletions have been reported in multiple countries in Africa, reaching very high frequency at the Horn of Africa [hrp2 deletion is a high-priority task of the WHO [The use of RDTs is threatened by f Africa . Molecul the WHO , and is  the WHO .kdr mutations increased from 1 to 86% [The success of LLIN distribution campaigns and IRS, which was recently scaled up in Burundi, crucially depends on killing of mosquitos by the insecticides used. Growing levels of insecticide resistance are a thread throughout Africa . In resp1 to 86% . New stuBurundi did well in containing the spread of SARS-CoV-2 in the first months of the pandemic. Border closures were the most important intervention. Only 818 confirmed cases were reported until December 31st, 2020. Since then, cases numbers have increased to over 4700 as of May 31, 2021. The pandemic did not interrupt malaria control interventions, i.e. IRS and LLIN distribution. Vector control and case management interventions were adapted as per WHO guidelines to mitigate the impact of covid-19 on delivery of HIV, tuberculosis, and malaria control services . The panThe steadily increasing number of malaria cases in Burundi since 2000 was observed despite a large increase in the number of health facilities, number of tests conducted, number of bed nets distributed, changes in treatment guidelines, and more recently introduction of IRS and IPTp. Despite a 20-fold increase in testing, the test positivity rate remained stable at around 50–60%. Given low testing numbers in the early phase of the study period, case numbers were likely underestimated. The consistently high positivity rate indicates that still many cases are missed.hrp2 deletion is recommended.In order to reduce the burden of malaria in Burundi, existing control activities need to be scaled up. New interventions that are currently tested in different settings might aide malaria control in the future, e.g. novel vector control tools such as larviciding , or acti


The aim of this study was to predict the progression-free survival (PFS) in patients with small cell lung cancer (SCLC) by radiomic signature from the contrast-enhanced computed tomography (CT).A total of 186 cases with pathological confirmed small cell lung cancer were retrospectively assembled. First, 1,218 radiomic features were automatically extracted from tumor region of interests (ROIs) on the lung window and mediastinal window, respectively. Then, the prognostic and robust features were selected by machine learning methods, such as (1) univariate analysis based on a Cox proportional hazard (CPH) model, (2) redundancy removing using the variance inflation factor (VIF), and (3) multivariate importance analysis based on random survival forests (RSF). Finally, PFS predictive models were established based on RSF, and their performances were evaluated using the concordance index (C-index) and the cumulative/dynamic area under the curve (C/D AUC).In total, 11 radiomic features  were finally selected, and the predictive model constructed from them achieved a C-index of 0.7531 and a mean C/D AUC of 0.8487 on the independent test set, better than the predictions by single clinical features , and single radiomic features computed in lung window  or mediastinal window .The radiomic features computed from tumor ROIs on both lung window and mediastinal window can predict the PFS for patients with SCLC by a high accuracy, which could be used as a useful tool to support the personalized clinical decision for the diagnosis and patient management of patients with SCLC. Small cell lung cancer (SCLC) is an aggressive pulmonary neuroendocrine tumor that causes about 250,000 deaths worldwide yearly , 2. SCLCIn clinical practice, CT imaging is widely used for the clinical diagnosis of SCLC. However, CT-based SCLC diagnosis greatly depends on the experience and knowledge of radiologists. They can provide some qualitative analysis for SCLC prognosis by their experience, but the quantitative analysis of SCLC prognosis is generally lacked . In the Recently, radiomics analysis is increasingly popular in tumor field, which is used for the diagnosis, stage, histology, prognosis, and treatment response assessment of various tumors , 13. By We retrospectively search the database from November 2012 to May 2019 under an active institutional review board, and written informed consents were acquired. The inclusion criteria were as follows: (1) patients pathologically proved SCLC by biopsy or operation. (2) Patients underwent chest CT scans. (3) Patients received standard Etoposide–Cisplatin combination chemotherapy. The exclusion criteria were as follows: (1) patients with treatment history before baseline CT scans; (2) patients lack of stage before treatment; (3) patients losing contact or die before progression; (4) patients without progression until deadline; (5) patients without contrast enhanced CT imaging; and (6) insufficient CT imaging. Finally, 186 cases were included in this study and divided into a training set (130 cases) and an independent test set (56 cases). The clinical information, such as age, gender, and stage were recorded.All patients underwent contrast-enhanced chest CT on 64-slice multidetector row CT scanner  with the following acquisition and reconstruction parameters: tube voltage of 120 kV; tube current of intelligent mAs; section thickness of 5 mm, and reconstruction interval of 5 mm. The contrast enhanced CT was administered intravenously with an amount of 60–70 ml of iohexol  followed by a saline flush of 20 ml, by using a power injector  at a flow rate of 2.5–3.0 ml/s.http://www.itksnap.org/pmwiki/pmwiki.php). Each tumor was segmented two times, first on the mediastinal window for consistency and then on the lung window, respectively. The lung window level (1600 and −500 HU) and the mediastinal window level (300 and 40 HU) were used during the tumor segmentation. The vessels and air regions were carefully excluded from the segmented tumor regions.A radiologist with 10 years of experience segmented each tumor region manually slice-by-slice on the axial CT images using the ITK-SNAP software  model . FeatureF is the unselected feature set, c(•) is the C-index of the CPH model which can be calculated by Equation 4. Second, to remove redundancy from image features, the variance inflation factor (VIF) was applied to quantify the collinearity between features  for multivariate analysis to further simplify the features . An RSF In above procedures, statsmodels , scikit-Based on the selected radiomic features and three clinical features , the random survival forests can be used to establish their corresponding prognostic model. For clinical features, selected lung window features and mediastinal window features, three different prognostic models can be generated, respectively. By combining the two classes of selected radiomic features into a feature set and simplifying them according to the Pearson's correlation , the cort-test and χ2 test were performed for continuous and categorical variables, respectively. The correlation matrix of selected features was calculated based on Pearson's correlation coefficient and illustrated in a heatmap. The predictive capacity of radiomic signature was evaluated using Kaplan–Meier analysis  curve (AUC) in survival data . All statistical analyses were two-sided with the statistical significance level of 0.05, and performed with statsmodels, scikit-learn, and scikit-survival packages in Python 3.6.Where, ωp = 0.334–0.771). In addition, through the univariate survival analysis, stage (p = 0.007) and sex (p = 0.039) were statistically relevant to the survival, and there was no significant difference regarding to age (p = 0.698).A total of 186 cases were enrolled and divided into a training set and an independent test set. As shown in During the follow-up, different types of progression were observed. Among 186 cases, there were 85 (45.7%) cases of primary progression, 23 (12.4%) cases of original metastases progression, 13 (7.0%) cases of primary progression and newly metastases, 11 (5.9%) cases of both the primary and metastasis progression, 16 (8.6%) cases of newly brain metastases, 14 (7.5%) cases of newly bone metastases, 8 (4.3%) cases of newly lung metastases, 6 (3.2%) cases of newly liver metastases, 1 (0.5%) case of cardiac metastases, 1 (0.5%) cases of pancreas metastases, 3 (1.6%) cases of adrenal metastases, and 5 (2.7%) cases of newly multiple metastases.In total, 1,218 radiomic features were extracted from CT images on the lung window and mediastinal window, respectively, and the feature selection was performed independently on both windows. After the univariate analysis, 724 and 895 features were remained, respectively. Then, with an iterative collinearity elimination based on VIF, a mass of redundant features was removed, resulting in 17 and 16 remained features for the lung window and mediastinal window, respectively. Finally, an RSF-based multivariate analysis was performed on the remaining features, and 6 features for the lung window  and 6 features for the mediastinal window  were selected as radiomic signature, as shown in Based on 3 clinical, 6 lung window, and 6 mediastinal window features, three basic prognostic models can be established, respectively, namely, Model_C3, Model_L6, and Model_M6. For these basic models, the optimal risk cut-off was determined using log-rank test on the training set, and patients in the training set and test set were stratified into high-risk and low-risk groups using the same cut-off, respectively. In By combining the lung window and mediastinal window radiomic features into a feature set, the combined model based on all 12 radiomic features (Model_L6+M6) was established. Additionally, to simplify radiomic features and reduce the risk of overfitting, we removed features with the higher correlation between two feature sets . Then, 11 remained radiomic features were used to generate a simplified model (Model_L5+M6). The overall prognostic ability of each prognostic model was evaluated with C-index and illustrated in The C/D AUC can evaluate the model performance from time level on the test set. The C/D AUC_90 evaluates how well these models can distinguish the patients progressing before and after 90 days, which is an important time for the prognosis of SCLC. As illustrated in Based on our survival models, a survival function can be generated and used to analyze the prognosis for each patient. Three typical cases with different survival time were illustrated in Two typical cases which are challenging and hard to predict the prognosis are illustrated in In this study, a PFS predictive model was proposed by integrating the radiomic features extracted from lung window and mediastinal window CT images in patients with SCLC. Our main findings indicated the contrast enhanced-mediastinal window radiomic features as an independent reliable prognostic factor. Our model effectively separated the groups of high risk and low risk, gave better predictive performance than the typical clinical visual analysis, and generate a survival function to analyze the prognosis for each patient. Our radiomics-based model offers more reliable PFS predictions which could support the personalized clinical decision-making.Radiomics can extract more quantitative information than bare eye to guide clinical decisions, usually from non-enhanced CT images. Unenhanced CT images reflect the tumor heterogeneity and microenvironment, demonstrating the prognostication and treatment response. Fried et al.  and GaneIn this study, we investigated the value of radiomic signature in predicting the prognostication and treatment response in patients with SCLC. Two cases with different prognosis are shown in Our radiomic models showed a good performance of C-index in predicting the PFS with conventional lung window features and enhanced mediastinal window features respectively, with a lowest C-index of clinical model. Furthermore, the predictive performance of radiomic features in an enhanced mediastinal window was superior to the lung window, indicating that enhanced mediastinal window features played an important role in predicting the PFS in patients with SCLC. Some previous studies showed that the predictive performance of quantitative texture features were similar between non-contrast and contrast-enhanced CT in diagnosing lung nodule , 44. AnoFor further verification, we analyzed the time-dependent cumulative/dynamic ROC curve and calculated the time-dependent AUC. The time-dependent cumulative/dynamic ROC curve analysis defined a marker value updated at each time point during the disease status individually, allowed to compare the marker's predictive ability and may give guidance for medical decisions . In our There were numerous texture features in CT images, which may provide different anatomical and biological information in tumor. Thus, the selection of texture features was meaningful and time-saving for model building. During feature extraction, original images were transformed to derived images with the wavelet filter and LoG filter to extract more radiomic features. After feature selection, the most important features for lung window and mediastinal window were selected respectively. For both lung window and mediastinal window, 50% (3/6) selected features were LoG-based, including the top-ranked features. These results illustrated that LoG-based features play a more important role in the survival analysis of SCLC. Actually, a LoG filter could enhance the edge information by emphasizing the areas of gray level change. The fine textures and coarse textures of the tumor were both taken into consideration by high and low sigma parameter. During the clinical diagnosis of SCLC based on CT images, the texture feature of the tumor edge is always an important indicator, which also proves the rationality of extracted radiomic features.There were several limitations in our study. First of all, due to the relatively low incidence of SCLC in Asian race, the population of this study is relatively small. Second, previous studies indicated that the suitable section thickness may be as thin as 1.25 or 2.5 mm. But the section thickness was 5 mm in this retrospective study, which may reduce the predictive accuracy of lung window. Third, smoking is an important risk factor in patients with SCLC especially in women, which did not calculate into the clinical features in our study.In summary, our study revealed that the textual features extracting from the contrast-enhanced mediastinal window were useful for predicting the PFS. The integration of textual features from the lung window and contrast-enhanced mediastinal window provided the more valuable information in survival prediction in comparison with the conventional visual assessment, which could be applied to support personalized clinical decision for the diagnosis and patient management in patients with SCLC.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.The studies involving human participants were reviewed and approved by Zhongshan Hospital of Fudan University. The patients/participants provided their written informed consent to participate in this study.XY and LW designed and conducted the study. NC, RL, XY, and LW contributed to the manuscript writing process. NC and RL prepared the first draft of the entire manuscript. NC performed tumor segmentation. RL, DS, and LW contributed to the radiomic feature extraction, feature selection, and data analysis. NC, MJ, YG, and JC contributed to data interpretation. All authors provided the final approval of the version submitted for publication.NC reports funding from the Medical and Health Foundation for Young Scientists of Fujian Province (Grant No. 2020QNA059).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Eichhornia crassipes leaves (LECs) in a packed-bed column was investigated by varying relevant operational parameters and assessment of mathematical models. Results showed that the zero-charge point of LECs was 2.37 and that optima pH and volumetric flux of the influent solution for AR27 biosorption were 2.0 and In this work, the biosorption behavior of acid red 27 (AR27) dye using  Textile wastewaters may contain between 1 and 2500 mg/L of toxic synthetic non-biodegradable dyes3, toxic heavy metals, salts, cleansers, and corrosive-alkaline chemical agents4. Large amounts of synthetic dyes are disposed in water bodies due to the absence or inefficacy of wastewater treatment methods3. These generate environmental and public health problems because synthetic dyes affect the aesthetic properties of water, as well as aquatic flora and fauna, and are also toxic, mutagenic, teratogenic, carcinogenic, and harmful to human health5.Dyes are widely used in the textile, food, pharmaceutical, plastic, cosmetics, photographic and paper industries, with the textile industry representing the largest consumer of dyes7, cosmetics, phenol-formaldehyde resins, and photographic images8. These industrial activities produce large amounts of wastewater contaminated with AR27. However, AR27 can be very hazardous to health, with effects that include respiratory problems, birth defects, allergies, tumors9, genotoxicity, cytotoxicity, embryotoxicity, mutagenicity, carcinogenicity10, and endocrine disruption9. This azo dye has been banned in the United States of America by the Food and Drug Administration (FDA) and its use is strictly controlled in many other countries12.Acid red 27 (AR27) is a highly water-soluble anionic azo dye that is used extensively in industries to color natural and synthetic textiles, and produce leather, paper, food, beverages, confectionery, pharmaceuticals13. Additionally, the complex chemical structure of AR27, which includes an azo group and conjugated benzene ring, renders it extremely recalcitrant to degradation, because of which it continues to persist in the environment14. Thus, it is crucial to remove AR27 from industrial effluents before their discharge into water bodies, to prevent it from having harmful effects on the environment and health.Furthermore, the discharge of AR27-containing effluents into water bodies reduces the penetration of sunlight, which affects the viability and photosynthetic process of aquatic plants, and deprives aquatic animals of the oxygen required for their vital functions16, biosorption18, photocatalytic degradation20, reverse osmosis, nanofiltration21, ion exchange8, chemical oxidation22, chemical precipitation, coagulation, and biodegradation21. Conventional biological methods for the treatment of dye-contaminated wastewaters are not effective for the removal of dyes21 and can generate secondary products, which are far more toxic than the dye itself24. Major drawbacks to advanced chemical oxidation, chemical coagulation and precipitation, and photocatalytic oxidation methods are their high cost, complexity, and the generation of hazardous wastes21. Ion-exchange technique uses very high-cost resins that are quickly exhausted and difficult to regenerate8. Membrane-based separations consume an elevated amount of energy and require high-cost investments21; in addition, membranes become fouled very easily, and have a limited lifespan8. Adsorption/biosorption appears to be a more viable and attractive technology for the removal of dyes from industrial effluents because of its unique properties, which include effectiveness, ease of operation, flexibility, adaptability, simplicity of design, cost-effectiveness, efficiency, efficacy, insensitivity to toxic pollutants, eco-friendliness21, and adsorbent/biosorbent regeneration18.Different approaches have been developed to achieve efficient removal of pollutants from industrial effluents, such as adsorption19, Eichhornia crassipes25, zeolitic imidazolate framework22, surfactant modified dull pink clay8, activated carbon web prepared from acrylic fibrous waste26, Pisum sativum peel, Arachis hypogaea shell27, and 28 as adsorbents/biosorbents. Batch systems are useful for the treatment of small volumes of effluents, and for determining the maximum dye adsorption/biosorption capacity of the adsorbent and effectiveness of the dye removal process in aqueous solutions30. However, the operational conditions for batch and continuous biosorption processes differ considerably; hence, the data collected in batch systems are commonly not applicable to industrial-scale continuous treatment systems31.AR27 adsorption/biosorption studies have only been conducted in batch systems using activated carbon prepared from coconut husk fiber, commercial activated carbon32. Furthermore, the experimental data obtained using laboratory packed-bed columns are useful for the design, scale-up, optimization, and operation of continuous wastewater treatment processes on a pilot and industrial scale30. In addition, the modeling of the dynamic behavior of packed-bed columns is essential for identifying the relevant factors and mechanisms of the biosorption process, predicting future changes under different operating conditions, and the successful design, scale-up, and optimization of the biosorption processes31.For the continuous treatment of industrial effluents on a large scale, it is preferable to use packed-bed column systems because of their high effectiveness, versatility, ease of use, low fixed and operating costs, and effluent qualityTo the best of our knowledge, no study has reported on the adsorption/biosorption of the toxic AR27 dye in continuous systems; therefore, there are no reports on the kinetic modeling of the AR27 adsorption/biosorption process in packed-bed column systems. This study is the first to address these research gaps.Eichhornia crassipes leaves (LECs) were reported to be among the most effective biosorbents available for the removal of AR27 in aqueous batch systems, because of their high capacity and rate of dye biosorption25. Characterization studies of AR27-unloaded LEC, AR27-loaded LEC, and AR27-desorbed LEC revealed that amide I and amide II functional groups, which are present in LEC proteins, are primarily responsible for the removal of AR27 from aqueous solutions by LECs13. Moreover, LECs can be reused in at least seven AR27 biosorption-desorption cycles in a batch system without resulting either in apparent physical change or damage to LECs or in a decrease in AR27 biosorption capacity and desorption efficiency or in loss of biosorbent9. Therefore, it is crucial to thoroughly investigate the ability of LECs to perform the continuous biosorption of AR27 in a packed-bed column system.Recently, In the present work, the particles and packed beds of LECs were characterized, and their potential for AR27 biosorption from aqueous solutions was evaluated using a continuous packed-bed column system. The influence of fundamental design parameters, such as the pH, AR27 concentration, volumetric flux of the influent AR27 solution, and LEC bed height on the performance of the packed-bed column used for AR27 biosorption was examined. In addition, a mathematical modeling of the AR27 biosorption dynamics in the LEC packed-bed column was performed, and the experimental data were compared with values predicted using different breakthrough curve models.Eichhornia crassipes plants were collected from Xochimilco canals, in the San Gregorio Atlapulco area , Mexico City, Mexico. The plant samples were identified by staff from the Herbarium (Dr. A. Novelo) of the National School of Biological Sciences, National Polytechnic Institute, Mexico City, Mexico (Collection ID: 2028).25 was recovered, and stored in screw-capped glass bottles for later use in biosorption experiments conducted in the packed-bed column reactor.The leaves were separated from the rest of the plant, washed with abundant quantities of tap water, followed by distilled water. The leaves were then cut into small pieces and oven-dried at A 2 g/L stock solution of AR27  were prepared using distilled water, and examined using a Zetasizer Nano-25 analyzer  at room temperature  method35.The LEC pore diameter and specific surface area were determined from nitrogen adsorption-desorption isotherms at 77 K using a Quantachrome ASiQwin gas sorption analyzer . The LEC particles were degassed with nitrogen at 373 K. The LEC specific surface area was evaluated using the Brunauer, Emmett and Teller (BET) multipoint method36.The porosity and bulk density values of the LEC packed bed were determined in triplicate, via the procedure described by AtarésContinuous AR27 biosorption experiments were performed in a 20-cm-high cylindrical glass column with an internal diameter of 1.3 cm  using silicone tubing. AR27 solutions with known concentrations and pHs were pumped up through the biosorbent bed at the desired flow rate, at room temperature , and an influent solution with a constant AR27 concentration of 50 mg/L, while ensuring that the volumetric flux of the AR27 solution was Likewise, four different volumetric fluxes, 37.67, 56.5, 75.34, and To investigate the effect of the inlet AR27 concentration on dye biosorption, experiments were performed using an influent solution with a volumetric flux of When the effect of the LEC bed height was examined, the volumetric flux of the AR27 solution was 13.A fraction collector  was used to collect effluent samples from the packed-bed column reactor, which were centrifuged at 3000 rpm for 10 min. Then, supernatants were analyzed using a Thermo Fisher Scientific Evolution 201 spectrophotometer at a wavelength of 520 nm to determine the AR27 concentrationControls were run without LEC biomass under the same operating conditions as those used in the continuous AR27 biosorption experiments, to detect possible dye losses due to AR27 adsorption on the glass, photolysis, and/or precipitation. The AR27 concentrations of the influent and effluent solutions did not show statistically significant differences in any of the LEC-free controls assayed, which indicates that the decrease in the AR27 concentration observed in the effluents from the continuous biosorption experiments with LEC biomass was solely attributable to the biosorbent.t). The specific biosorption capacity  of AR27  was calculated using Eq. 32:The continuous biosorption of AR27 on the LEC packed-bed column was studied in terms of the breakthrough curves, which were expressed by the relationship between the outlet AR27 concentration , modified Dose-Response, and logistic models were used in the present study to match the experimental data, and predict the dynamic behavior during the process of AR27 biosorption on LEC in the packed-bed column at different pHs, volumetric fluxes, AR27 concentrations of the influent solution, and LEC bed heights.30:The Thomas non-linear model has the following simplified form Eq. 30:3\docu39:The bed depth service time (BDST) model can be expressed in terms of Eq. 440:The non-linear form of the Yoon–Nelson model is expressed as follows Eq. 40:5\docu41:The modified dose-response model can be expressed in terms of Eq. 642:42.It has been claimed that the Bohart–Adams (variant of the BDST model), Thomas, and Yoon–Nelson models can be expressed in terms of the logistic model, which is expressed as follows Eq. 42:7\documentclass2pt{minimTo ensure result reproducibility and perform statistical analysis, continuous AR27 biosorption experiments were carried out in triplicate. The mean ± standard deviation values are reported in this work.GraphPad Prism version 8.0 software  was employed both to perform the statistical analysis of the AR27 biosorption data and determine the kinetic parameters of the packed-bed column models. The AR27 biosorption data were subjected to analysis of variance (ANOVA) and Tukey’s test with a 95% confidence level.The kinetic parameters of packed-bed column models were determined using non-linear regression analysis. The models that were best adjusted to the experimental data were selected, to ensure that the coefficient of determination . Likewise, the zeta potential became more negative as the pH of the solution increased from the zero-charge point pH to pH 10, which indicates that the net charge on the LEC surface became more negative as the solution pH increased. Therefore, the decrease observed in the AR27 biosorption capacity in solutions with pHs ranging from 3 to 6 was attributable to the fact that when the solution pH increased, the electrostatic repulsive forces between the negative charges on the anionic AR27 dye and negative charges on the LEC surface increasedIt is apparent from our results that the optimal pH for continuous AR27 biosorption was 2.0, since at this pH, the specific (67.25 mg/g) and volumetric  AR27 biosorption capacities were maximal, and the operating time of the packed-bed column was longer. Therefore, subsequent studies were carried out using an influent AR27 solution with a pH of 2.0.Figure Moringa oleifera seeds45, remazol reactive dyes onto Rhizopus arrhizus NCIM 99746, acid blue 25 onto NaOH-treated fallen Ficus racemosa leaves47, and reactive blue BF-5G onto malt bagasse48 in packed-bed columns.The above results clearly show that the service and saturation time were longer for the lowest volumetric flux tested and shorter for the highest volumetric flux of the influent AR27 solution assayed. Similar results were found previously for the biosorption of yellow dye tartrazine onto Similarly, the highest mass of removed AR27, and highest specific and volumetric biosorption capacities of AR27 were observed at volumetric fluxes of 37.67 44.The decrease in the service and saturation time, AR27 mass removed, and specific and volumetric biosorption capacity with an increase in the influent volumetric flux may be attributable to the fact that the contact time between the AR27 azo dye and the LEC biomass in the column decreases as the volumetric flux increases. Thus, the AR27 molecules do not have enough time to attach to the biosorption sites present on the LEC surface or diffuse into the LEC pores, and leave the packed-bed column at higher concentrations, before biosorption equilibrium can be attainedOur results show that at a volumetric flux of 56.5 Figure At influent AR27 concentrations of 30, 50, 75, 100, and 200 mg/L, the column effluents did not contain a detectable amount of AR27 dye during the first 60, 45, 30, 21 and 6 h of column operation, respectively, and all the dye initially present in the inlet solutions was biosorbed. In contrast, AR27 dye was found in the outflow effluent from the start of operation of the packed-bed column, when an inlet solution with an AR27 concentration of 400 mg/L was fed to the column. These results clearly show that the service time of the column increased with a decrease in the influent AR27 concentration.Similarly, the slope of the breakthrough curves increased as the concentration of influent AR27 increased, which resulted in a decrease in column saturation time with an increase in the influent AR27 concentration.32.It is evident that both the service time and saturation time of the packed-bed column were lower at higher influent AR27 concentrations. This can be explained by the fact that the higher the influent AR27 concentration, the greater the AR27 concentration gradient, and consequently, the greater the thermodynamic driving force for dye biosorption. Thus, the AR27 molecules are biosorbed more quickly onto the biosorbent when the dye concentration is higher; consequently, the service and saturation time periods are minimalEnteromorpha prolifera49 , Eucalyptus sheathiana bark50, and pyrolytic tire char30, crystal violet by Citrullus lanatus rind and Cyperus rotundus51, direct blue 71 by cross-linked chitosan-glutaraldehyde beads29, and reactive red 198 by activated carbon developed from walnut shells52.Qualitatively similar behavior was reported in previous packed-bed column studies on the biosorption of methylene blue by 25, and is considerably higher than those reported in a few studies on batch AR27 adsorption, including those using peanut hull53, de-oiled soya, bottom ash54, raw and 55, alumina reinforced polystyrene24, dull pink clay8, polyethyleneimine grafted chitosan beads14, activated carbon web produced from acrylic fibrous waste26, 28, and Arachis hypogaea shells27 as adsorbents.The maximum specific and volumetric biosorption capacities of AR27 were observed at inlet AR27 concentrations ranging from 50 to 400 mg/L  model) are the most commonly used mathematical models for the analysis of the dynamic behavior of the packed-bed column in the adsorbate-adsorbent system. It has been argued that all these models can be represented using the logistic model of population growth with two general parametersIn this work, the BDST, Thomas, modified dose-response, Yoon–Nelson, and logistic models were adjusted to the experimental breakthrough data to describe the dynamic performance of the packed-bed column reactor at various pHs, volumetric fluxes, AR27 concentrations and LEC bed heights.Tables 50.The rate constant The q values predicted by the Thomas model were higher at solution pHs of 1.5 and 2.0 and at volumetric fluxes of 37.67 and 56.5 30.Likewise, all LEC bed heights assayed showed q values not to manifest significant differences and nor did influent AR27 concentrations ranging between 50 and 400 mg/L. The Thomas model is apt for describing biosorption processes, where external and internal resistance to the mass transfer of solute from the liquid to the solid phase is extremely low, and likewise biosorption is not limited by the chemical reaction, but is controlled by mass transfer at the interface50. The 50. The volumetric biosorption capacity 51.The rate constant (58, Han et al.59, Afroze et al.50, and Chen et al.60. The Yoon–Nelson model assumes that the decline in probability for each adsorbate to be biosorbed is proportional to the probability of its biosorption and breakthrough on the biosorbent37.The 50% breakthrough time 37.The kinetic constants of the modified dose-response model (The kinetic constant a of the logistic model was higher at influent solution pHs of 1.5 and 2.0 but decreased as solution pH increased from 2.0 to 6.0. The a constant also decreased, when both the volumetric flux and the influent concentration were increased, but increased as the bed height increased. No significant difference was observed in the values for the b constant of the logistic model within a pH range for the influent solution of 1.5 and 4.0, but increased as solution pH increased from 4.0 to 6.0, as well as when volumetric flux, influent concentration, and bed height were increased.The above results clearly indicate that lower influent solution pHs, lower-middle influent volumetric fluxes, middle-higher influent AR27 concentrations and higher bed heights would increase the mass of AR27 dye biosorbed on the LEC packed-bed.42.It was also found that the BDST, Thomas, and Yoon–Nelson models had the same fitting effect as the logistic model, and were therefore mathematically equivalent. Similarly, it was observed that these four models and the modified dose-response model exhibited values of entclass1pt{minimay Tables , 4, 5. IThe above results indicate that the Thomas, BDST, Yoon–Nelson, logistic, and modified dose-response models were able to represent the behavior of the breakthrough curves for AR27 biosorption by the LEC packed bed.45 also found that the Thomas, Adams-Bohart, and dose-response kinetic models were able to reproduce the experimental data of tartrazine biosorption in a packed-bed column. Experimental results and mathematical modeling suggest that internal and external diffusion are not the controlling steps in the AR27 biosorption by LEC but is controlled by the mass transfer at the interface.Reck et al. In the present study, the LEC particles were characterized in terms of average specific surface area, particle size, pore size, zeta potential, and zero-charge point. Likewise, the porosity and bulk density of LEC packed-bed were determined. The biosorptive effectiveness of sustainable and cost-effective LEC biomass in the biosorption of AR27 from aqueous solutions was assessed using a continuous up-flow packed-bed column reactor. The AR27 biosorption in the continuous biosorption system depends on the pH, volumetric flux, and AR27 concentration of the influent AR27 solution, as well as on the LEC bed height. The packed-bed biosorption system was found to perform better when operated with an influent solution pH of 2.0, a volumetric flux of inlet solution of 56.5


CAV1, PACSIN2, and CDC42, which are the target genes of lncRNAs in the same profile, suggesting the possible regulatory relationship between lncRNAs and mRNAs. The current study reveals the expression profiles and functional networks on messenger RNAs (mRNAs) and long non-coding RNAs (lncRNAs) in the liver of Ningxiang piglets across four developmental stages . Differentially expressed mRNAs (DEmRNAs) were upregulated at 30 days; however, most differentially expressed lncRNAs (DElncRNAs) were downregulated at 210 days. A complex interaction between mRNAs and lncRNAs was identified by Short Time-series Expression Miner (STEM) analysis and weighted gene co-expression network analysis (WGCNA), indicating that lncRNAs may be a critical regulatory element in mRNAs. STEM was used to identify significant temporal expression profiles and the genes associated with them and to compare the behavior of these genes across multiple conditions. WGCNA was used to study the biological networks based on pairwise correlations between variables. One particular mRNA profile 4 contained CAV1, PACSIN2, and CDC42 in the particular mRNA profile 4 were the target genes of lncRNAs in that profile, which shows the possible regulatory relationship between lncRNAs and mRNAs. The expression of these genes and lncRNAs in profile 4 was the highest at 30 days, and it is believed that these RNAs may play a critical role during the suckling period in order to meet the dietary requirements of piglets. In the lncRNA–mRNA co-expression network, the identified gene hubs and associated lncRNAs were shown to be involved in saccharide, lipid, and glucose metabolism, which may play an important role in the development and health of the liver. This result will lead to further investigation of liver lncRNA functions at various stages of development in Ningxiang pigs.Ningxiang pigs, a fat-type pig, are native to Ningxiang County in Hunan Province, with thousands of years of breeding history. This study aims to explore the expression profiles and functional networks on messenger RNAs (mRNAs) and long non-coding RNAs (lncRNAs) in the liver. Liver tissue of Ningxiang piglets was collected at 30, 90, 150, and 210 days after birth (four development stages), and the mRNA and lncRNA expression was profiled. Compared to mRNA and lncRNA expression profiles, most differentially expressed mRNAs (DEmRNAs) were upregulated at 30 days; however, most DElncRNAs were downregulated at 210 days. Via Short Time-series Expression Miner (STEM) analysis and weighted gene co-expression network analysis (WGCNA), a complex interaction between mRNAs and lncRNAs was identified, indicating that lncRNAs may be a critical regulatory element for mRNAs. One module of genes in particular (module profile 4) was related to fibril organization, vasculogenesis, GTPase activator activity, and regulation of kinase activity. The mRNAs and lncRNAs in module profile 4 had a similar pattern of expression, indicating that they have functional and regulatory relationships. Only  There are about 20,000 protein-coding genes in the human genome, accounting for less than 2% of the whole genome . Those sNingxiang pigs, a fat-type pig, are native to Ningxiang County in Hunan Province, with thousands of years of breeding history. In the long-term breeding process, Ningxiang pigs, one of the four famous pig species in China, are popular for their advantages, including tender and succulent meat, unique flavor, and high-quality unsaturated fatty acids, as compared to other local pig species. Ningxiang pig is an important national livestock resource and was approved by the Ministry of Agriculture in 2010 as a geographical representation of Chinese agricultural products . In the The liver, the primary digestive organ in mammals, is involved in the metabolism of amino acids, carbohydrates, lipids, and other substances by secreting bile, digestive enzymes, and hormones. The development and functions of the liver take place from birth. Through the development and maturation of all kinds of functions, the liver plays an essential role in digestion and metabolism ,11. ThisIn this research, we used high-throughput sequencing technology to systematically identify the lncRNAs and mRNAs of the liver at four developmental stages . Interestingly, 30 days after birth was a significant period for lipid biosynthesis in the liver. STEM and WGCNA showed that mRNAs and lncRNAs were enriched with lipid, saccharide, and protein metabolism, and other biological processes, and they may play an important role in liver development, health, and tissue repair. In the meantime, the potential regulatory relationship between lncRNAs and mRNAs has been further confirmed. The results from this study can provide references for further study of the function and mechanisms of lncRNAs in the liver and may also provide an important reference for the research of other Chinese pig breeds. The findings from this study provide molecular basis for subsequent research on Ningxiang pigs, such as the next lncRNA function study to provide candidate genes, and at the same time understand the development characteristic of Ningxiang pigs in actual production and provide an important reference for the research on nutrition and feeding management of Ningxiang pigs. A Ningxiang boar was mated with 4 Ningxiang sows to obtain half-sibling piglets. A total of 12 tails of half-sibling Ningxiang piglets were provided by the Ningxiang original breeding pig farm of Hunan Chuweixiang Agriculture and Animal Husbandry Co. Ltd. . Twelve healthy male full-sibling piglets with similar weights were randomly chosen to be slaughtered to collect liver samples at 4 development stages , and 3 tails were randomly chosen for each stage. All experimental pigs were fed, reared and managed in the same manner and condition from birth. During the liver samples collection, all the liver tissue samples were collected from the same site in each animal at each collection time-point. The collected liver samples were immediately placed in liquid nitrogen, and then transferred to a freezer at −80 °C for RNA extraction. All animal experiments in this study were approved by the Institutional Animal Care and Use Committee of Hunan Agricultural University, Changsha, Hunan Province, China under approval number 2013-06. Total RNA from liver was isolated by TRIzol Reagent , following the manufacturer’s procedure. The isolated RNAs were treated with RNase-free DNase to eliminate excess DNA. The quality of isolated RNAs was assessed by a Nanodrop 2000  and 1% agarose gel electrophoresis. The qualified RNAs were kept in a freezer at −80 °C until use. An RNA-seq transcriptome strand library was prepared with a TruSeqTM stranded total RNA Kit from Illumina  using 5 μg of total RNA. Ribosomal RNA (rRNA) depletion instead of poly(A) purification was performed by a Ribo-Zero Magnetic kit and then fragmented by fragmentation buffer. The first-stranded cDNA was synthesized with random hexamer primers. The RNA template was removed to synthesize a replacement strand, incorporating dUTP instead of dTTP to produce double-strand cDNA (dscDNA). AMPure XP beads were used to isolate the dscDNA from the second strand reaction mix. A single A nucleotide was added to the 3’ ends of these blunt fragments to prevent them from ligating to each other during the adaptor ligation reaction. Finally, multiple indexing adapters were ligated to the ends of the dscDNAs. Libraries were size selected for cDNA target fragments with 200–300 bp on 2% Low Range Ultra Agarose, followed by PCR amplified using Phusion DNA polymerase  for 15 PCR cycles. After quantification by TBS380, a paired-end RNA-seq sequencing library was sequenced with the Illumina HiSeqxten (2 × 150 bp read length). https://github.com/jstjohn/SeqPrep, accessed on 5 October 2020) and Sickle  was used to assess and trim the sequencing read quality. The linker sequence in the reads, and the read with no inserted fragments due to self-connection of the linker, were removed; low-quality  at the end of the 3’ end sequence was trimmed off; the quality value less than 10 in the remaining sequence was removed; the reads with more than 10% of N was removed; the adapter and sequence whose length is less than 20 bp after quality trimming was discarded. The base content distribution assessment is used to detect the presence or absence of AT and GC separation. We mapped the sequence by using TopHat2 [Since the original sequencing data will contain sequencing adapter sequences, low-quality reads, sequences with a higher N rate, and sequences with too short length will seriously affect the quality of subsequent analysis. To ensure the quality and accuracy of subsequent biological information analysis, the original sequencing data is first filtered to obtain high quality clean data. The SeqPrep  was used to measure gene abundance. The 30-day vs. 90-day, 30-day vs. 150-day, 30-day vs. 210-day, 90-day vs. 150-day, 90-day vs. 210-day, and 150-day vs. 210-day groups were undertaken to compare the differential expression. Empirical Analysis of Digital Gene Expression in R (edgeR) software in the R statistical package was used for the analysis of the differential expression. In addition, a functional enrichment analysis, including Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) was performed to determine which DEGs were significantly enriched in GO terms and metabolic pathways at a Bonferroni-corrected p-value ≤ 0.05 compared to the whole transcriptome background. The GO functional enrichment and KEGG pathway analysis were carried out by Goatools and KOBAS. The adapter sequences were removed from the raw sequencing reads prior to alignment . The expression level of each transcript was determined based on fragments per kilobase of exon per million mapped reads (FRKM) for the identification of differential expression genes (DEGs) among the samples tested. RSEM . Model profiles with color (except white) indicate that the temporal trends of mRNAs and lncRNAs were statistically significant. Profiles that had the same color were grouped into the same cluster.The Short Time-series Expression Miner (STEM) clustering algorithm was used to classify protein coding genes and lncRNA expression profiles in order to explore the relationship between temporal gene expression patterns and the liver during the four stages of piglet development . The novThe mRNA–lncRNA co-expression networks were developed through the developmental phases of the weighted correlation network analysis (WGCNA) package of R software . After e−ΔΔCt method. Total RNA was extracted using an Animal Total RNA Kit  and treated with ribonuclease R to validate the identified lncRNAs in Ningxiang pig. cDNA was synthesized by reverse transcription using the Revert Aid First Strand cDNA Synthesis Kit . The transcription levels of MSTRG.1053.3, MSTRG.11451.1, MSTRG.19861.1, MSTRG.8339.1, and MSTRG.1054.2 were validated by Quanstudio 6 Flex . The forward and reverse primers for gene quantification are listed in After lncRNA and mRNA analysis of Ningxiang pig liver at 30, 90, 150, and 210 days after birth, 333,589,902, 328,613,762, 266,368,076, and 306,539,556 clean reads were obtained with more than 95.10% of Q30, respectively . These cAccording to the expression profiles of mRNAs, we found that the samples could be distinguished among the four development stages. Surprisingly, the 90-, 150-, and 210-day-old pigs had some similarities in the expression profiles, but the 30-day-old pigs showed the biggest differences from the other three age groups A. At 210ACSL3, CES1, CYP2C42, CYP4A24, and PLIN4 were associated with fatty acid metabolism, indicating that it is an important part of liver development   and 1158elopment . Enrichmelopment . The comCPT1B targeting  among the 13 module profiles, accounting for 80.98% of the total genes A–C. MED2n module A; TBPL1 t module B; and MOs module C. The tas module .Five lncRNAs—MSTRG.1053.3, MSTRG.11451.1, MSTRG.8339.1, MSTRG.10861.1, and MSTRG.1054.2—were randomly selected from the novel lncRNAs and quantified by RT-qPCR at four development stages . The results showed a concordance between the RNA-seq and RT-qPCR data, suggesting that the RNA-seq data was reliable .It has been widely recognized that lncRNAs play a significant role in tissue and organ development and metabolism regulation, and lncRNAs in the liver may influence digestion and metabolism. In this study, many differentially expressed mRNAs (DEmRNAs) and differential expressed lncRNAs (DElncRNAs) were identified. In the DEmRNAs study, the greatest differences occurred at 30 days compared to the other stages. This phenomenon suggests that the expression profiles at 30 days may be critical in liver development. Variations in the dietary structure during the suckling and weaning periods may affect the digestive function, which may be correlated with different expressions of mRNAs ,26. HoweACSL3, CES1, CYP2C42, CYP4A24, and PLIN4 were associated with fatty acid metabolism in common DEmRNAs. In the common DElncRNAs, MSTRG.31876.3 and MSTRG.33238.1 showed the highest expression levels at 150 days after birth. MSTRG.31876.3 was predicted to PCF11 and AGO1, which are primarily involved in transcriptional regulation; and MSTRG.33238.1 was predicted to several target genes: CDHR5, CYP4A24, HSD17B3, and TICAM1, among which CYP4A24 encoded for cytochrome p450. CYP4A24, also known as fatty acid omega-hydroxylase, regulates omega- and (omega-1)-hydroxylation of various fatty acids [TICAM1, encoding for TLR domain-containing adapter molecule 1, actively participates in innate immunity against invasive pathogens to protect the host [Venn diagrams were constructed to classify the common DEmRNAs and DElncRNAs along the four stages of development. ty acids ,28. TICAthe host . The finAPOA1, encodes for apolipoprotein A1 as part of the high-density lipoprotein (HDL) used to transport fat molecules back to the liver for excretion, had higher expression levels at 90, 150, and 210 days compared to 30 days after birth. APOA1 has been reported to be regulated by APOA1-AS, an antisense lncRNA, as a negative transcriptional regulator [APOA4, APOM, and ABCG were involved in the transport of lipoprotein particles and cholesterol. In addition, the KEGG pathway study showed that these genes were predominantly involved in lipid-related pathways, such as arachidonic acid metabolism, fatty acid biosynthesis, fatty acid metabolism, and PPAR signaling pathway. STEM analysis found that there was a dynamic and similar pattern of expression between mRNAs and lncRNAs during the development process in the liver. STEM analysis in this study revealed that mRNA profiles 21, 24, and 25 belonged to the same cluster, which was enriched for sterol and steroid biosynthesis and transport, triglyceride catabolism, lipoprotein particles, and cholesterol transport and esterification. In profile 21, Vezf1 of lncRNA in lncRNA profile 4 was found to encode the zinc finger transcription factor, which is transcribed in the endothelial cells during angiogenesis development and regulation, suggesting that Vezf1 may promote liver development and tissue repair [CPT1B, encoding carnitine palmitoyltransferase 1B, a member of the carnitine acetyltransferase family, functions in regulating long-chain fatty acid beta-oxidation by transporting long-chain fatty acyl-CoAs from cytoplasm to mitochondria [The genes in mRNA profile 4 were mainly enriched for fibril organization, vasculogenesis, GTPase activator activity, and regulation of kinase activity. The lncRNAs in profile 4 were primarily involved in long-chain fatty acids, membrane rafts, angiogenesis, and identical protein binding. Angiogenesis was also seen to have a key role in tumor growth and metastasis. A previous study reported that when the liver is damaged, regeneration of the liver is associated with angiogenesis, or is at least partially dependent on angiogenesis . Target e repair . In addichondria . The expCAV1, PACSIN2, and Cdc42 in a particular mRNA profile 4 were exactly the target genes of lncRNAs in lncRNA profile 4. A previous study found that CAV1 encoded for caveolin-1 protein, a scaffolding protein in caveolar membranes, which is primarily involved in endocytosis [CAV1 and PACSIN2 interact. PACSIN2 encodes the protein kinase C and casein kinase substrate mediating the shape formation of caveolae in plasma membranes, suggesting that PACSIN2 is a fundamental protein involved in cellular homeostasis and disease via caveolae regulation [CDC42, encoding a small GTPase-associated plasma membrane and identified as the Rho subfamily, participates in cellular responses, epithelial cell polarization processes [ocytosis , cell trocytosis , intraceocytosis , and lipocytosis , which mocytosis . Variousgulation . CDC42, rocesses , cell mirocesses , regulatMED24, ANO6, and ZC4H2 were identified as hub genes in the molecular function module. MED24, encoding mediator complex subunit 24 (TRAP100), a transcriptional coactivator complex involved in the induced transcription of almost all genes dependent on RNA polymerase II [TBPL1 was classified as a hub gene. TBPL1, encoding TATA box-binding protein-like 1, also known as TLF and TRF2, belongs to the TATA binding protein (TBP) paralog despite lacking the ability to bind TATA elements and interact with transcription factor II A and B, mediating the transcription of most ribosomal proteins to regulate gene transcription in development and differentiation. In the lncRNA–mRNA co-expression network, erase II , has severase II . In the MOGS, ACADSB, and DNAJC25 were the hub genes in the biological process module. MOGS encode mannosyl-oligosaccharide glucosidase isoform 1, which removes the distal α1,2-linked glucose residues from Glc(3)-Man(9)-GlcNAc(2) oligosaccharide precursor. MOGS is substantially involved in glucosidase activity, mannosyl-oligosaccharide glucosidase activity, oligosaccharide metabolic process, and protein N-linked glysosylation [MOGS mutation could lead to congenital glycosylation disorders (CDGs), which could lead to retarded development, low immunoglobulin levels, and abnormal liver function [ACADSB encodes a short/branched chain specific acyl-CoA dehydrogenase (SBCAD), known as 2-methylbutyryl-CoA dehydrogenase (MBD), which catalyzes the first step of L-2-methylate short acyl-CoA compounds in the mitochondria [ACADSB also functions in inisoleucine catabolism, fatty acid β-oxidation, and lipid metabolism [DNAJC25 encodes DNAJC homologous subfamily C member 25, known as HSP40 homolog, which binds to chaperon HSP70 by its J domain to facilitate ATP hydrolysis for protein folding, unfolding, translation, and degradation, and had particularly higher expression in the liver than adjacent normal tissues [sylation . A previfunction . ACADSB chondria . ACADSB tabolism . DNAJC25HSP70 has been reported to reduce liver injury and promote liver health during the weaning period [HSP40 may interact with HSP70 and they may function together in a variety of physiological processes, suggesting that HSP40/DNAJC25 could play an important role in liver health, particularly when under stress or injured. This obviously shows that the co-expression network of protein coding genes and lncRNAs is significantly involved in liver protection. The liver is important in maintaining the heath and growth of an animal. If the liver is injured, the homeostasis will be harmed. We can observe that liver protection is important during the early stage of development, which might be due to the incomplete development of the immune system. It is necessary to protect the liver in order to ensure the survival of piglets. During the development of piglets, the liver is often injured by many factors, including weaning stress and digestive tract diseases . Increasg period . HSP40 mCAV1, PACSIN2, and CDC42 in the particular mRNA profile 4 were only the target genes of lncRNAs in that profile, which suggests a possible regulatory relationship between lncRNAs and mRNAs. The expression of these genes and lncRNAs in profile 4 was the highest at 30 days, indicating that these RNAs may play a critical role during the suckling period to meet the dietary requirements of piglets. The findings of the current study lay a foundation for the study of lncRNAs in Ningxiang pigs and offer new insights for lncRNA functions in the liver at various stages of development. Research on the function of detected DEmRNAs and DElncRNAs should be carried out in order to gain deeper knowledge of the role and regulation of these DEmRNAs and DElncRNAs. A deeper understanding of these genes may help producers to manipulate or enhance the production of pigs.In conclusion, the present study reveals the expression profiles and functional networks on mRNAs and lncRNAs in the liver of Ningxiang piglets across four developmental stages . We found that most of the DEmRNAs were upregulated at 30 days, but most were downregulated at 210 days. A complex interaction between mRNAs and lncRNAs was identified, indicating that lncRNAs may be a critical regulatory element in mRNAs according to STEM and WGCNA analysis. mRNAs and lncRNAs have similar patterns of expression in module profile 4, indicating that they have functional and regulatory relationships. Module profile 4 is related to fibril organization, vasculogenesis, GTPase activator activity, and regulation of kinase activity. We must point out that