The obtained results indicated that size and especially polarity were negatively correlated with the diffusion capacity. Although some antifungal compounds, such as prochloraz and carbendazim, were completely degraded after a few days in peel and mesocarp, other compounds, such as p-aminobenzoic acid and kojic acid, showed high stability. When applying the antifungal compounds at the EC50 concentrations, it was observed that the compounds with high diffusion capacity showed higher in vivo antifungal activity against Alternaria alternata than compounds with low diffusion capacity. In contrast, there was no relationship between stability and in vivo efficacy. Collectively, the obtained results indicated that the diffusion capacity plays an important role in the efficacy of fungicides for the control of pear fruit diseases.Fungal pathogens can invade not only the fruit peel but also the outer part of the fruit mesocarp, limiting the efficacy of fungicides. In this study, the relationships between fungicide structure, diffusion capacity and in vivo efficacy were evaluated for the first time. The diffusion capacity from pear peel to mesocarp of 11 antifungal compounds, including  Pyrus spp.) are one of the most consumed fruits worldwide. The production of pears reached almost 24 million tons in 2019, with main production areas in China, Italy and the USA  × 100, where T is the diameter of the lesion length for the treatment sets, and C is the diameter of the lesion length for the control sets [A. alternata caused brown lesions on the fruit peel around the inoculation site that were observed with the naked eye. Twenty-five pears were used for each treatment condition. Experiments were repeated three times.The antifungal efficacies for the control of rol sets . A. altep ≤ 0.05. ANOVA and linear regression analyses were performed using the SPSS software (version 16.0).The obtained data were analyzed by the one-way ANOVA method and the significance levels were calculated by Tukey’s multiple range test. Differences between means were considered significant when p-aminobenzoic acid, carbendazim, difenoconazole, dipicolinic acid, flusilazole, gentamicin, kojic acid, prochloraz, quinolinic acid, thiophanate methyl and thiram were used in this study. Carbendazim, difenoconazole, flusilazole, prochloraz, thiophanate methyl and thiram have been commercialized for agricultural uses. The antifungal properties of p-aminobenzoic acid, dipicolinic acid, gentamicin, kojic acid and quinolinic acid against some plant pathogens have been examined; however, these compounds are only experimental and have never been commercialized for agricultural use. When selecting molecules for the purpose of this study, compounds with a variety of structural characteristics were prioritized to show how these structural differences influence diffusion capacity, stability and, thus, in vivo efficacy. The effects of the structural characteristics of fungicides on their in vitro antifungal activities have been thoroughly studied, and several structure/activity relationship studies can be found in the literature [Eleven fungicides, including terature ,40. Howeterature .p-aminobenzoic acid, carbendazim and quinolinic acid . It must be noted that these compounds showed some of the lowest volumes  among all studied compounds, suggesting that small compounds show higher diffusion capacities in pear peel compared to large compounds. Accordingly, gentamicin and flusilazole, which have large volumes , were only detected in the pear peel but not in the pear mesocarp, indicating that they are unable to penetrate the peel to reach the internal phases of the pear fruit. The minimum energy conformations of the studied fungicides are shown in As can be seen in p-aminobenzoic acid and quinolinic acid showed high diffusion capacities , whereas the diffusion capacities of dipicolinic acid and kojic acid were much lower . As shown in p-aminobenzoic acid and quinolinic acid  were lower than those calculated for dipicolinic acid and kojic acid , suggesting that the polarity of the compounds also affects the diffusion capacity, with nonpolar compounds showing higher diffusion capacities than polar compounds.When comparing compounds with small volumes, it can be observed that other factors affect diffusion. For example, p < 0.001) and size (p < 0.001) having significant effects and being negatively correlated with the diffusion capacity  nor polarity (p > 0.05) were significantly correlated to the stability of the compounds , followed by flusilazole (EC50 = 0.51 mM), thiram (EC50 = 0.52 mM), difenoconazole (EC50 = 0.79 mM), dipicolinic acid (EC50 = 4.5 mM), gentamicin (EC50 = 5.3 mM), thiophanate methyl (EC50 = 11.1 mM) and p-aminobenzoic acid (EC50 = 11.4 mM). On the other hand, carbendazim (EC50 = 20.1 mM), kojic acid (EC50 = 20.3 mM) and quinolinic acid (EC50 = 46.9 mM) showed the highest EC50 values . Although Vujanovic et al. reported that 4.16 mM thiram only inhibited the mycelial growth of an A. alternata (using a strain isolated from asparagus) by 25% [50 of thiram to A. alternata HN-5 was 0.52 mM, indicating that HN-5 is more sensitive than the other strain. As far as we know, this is the first study on the antifungal activities of dipicolinic acid, flusilazole, gentamicin, p-aminobenzoic acid, kojic acid and quinolinic acid to A. alternata.In agreement with the ECuritiana . Severalted here ,57. The recently ,59. In aersimmon . The EC5 1.13 mM , which a) by 25% , the EC5p-aminobenzoic acid, carbendazim and quinolinic acid strongly inhibited A. alternata symptoms ; however, the stabilities in peel (p > 0.05) and mesocarp (p > 0.05) did not significantly affect the antifungal efficacy. The regression equation was: antifungal efficacy = −0.001 + 1.941 × diffusion capacity (To confirm the correlations between diffusion/stability and in vivo antifungal activity, a linear regression analysis was carried out. The obtained results showed that diffusion influenced the in vivo activity by 66.2% (capacity B.Alternaria alternata in pear, it must be noted that the presence of toxic fungicides in the mesocarp poses a food safety hazard. Although the removal of the fungicides from the fruit peel can be done easily by peeling with a knife, the removal of the fungicides from the mesocarp seems a difficult task. Some of the studied fungicides are known to cause negative effects on human health. For example, carbendazim has been related to cancer, and is known to produce infertility, hepatocellular dysfunction and developmental toxicity in mammals [Although fungicides with high diffusion capacity allowed the control of  mammals . Difenoc mammals ,65,66, w mammals ,68.A. alternata was carried out just after the treatment. The stability must be involved in the in vivo efficacy when considering different inoculation time points, and the stable compounds may be more efficient than the unstable compounds for long periods.Our results suggested that the stability was not correlated with the in vivo antifungal activity; however, the inoculation of In summary, this study has revealed for the first time that size and polarity play an important role in the diffusion capacity of fungicides in pear fruit, with small and non-polar fungicides showing higher diffusion capacity than those with large size and high polarity. At the same time, the diffusion capacity was found to be positively correlated with the in vivo antifungal activity of the fungicides. This study provides new insights regarding the structural characteristics that are suitable for the control of fungal pathogens in pear fruit and will lead to a better understanding of the factors involved in the in vivo efficacy of fungicides.


Water contamination is one of the most urgent concerns confronting the world today. Heavy metal poisoning of aquatic systems has piqued the interest of various researchers due to the high toxicity and carcinogenic consequences it has on living organisms. Due to their exceptional attributes such as strong reactivity, huge surface area, and outstanding mechanical properties, nanomaterials are being produced and employed in water treatment. In this review, recent advances in the use of nanomaterials in nanoadsorptive membrane systems for wastewater treatment and heavy metal removal are extensively discussed. These materials include carbon-based nanostructures, metal nanoparticles, metal oxide nanoparticles, nanocomposites, and layered double hydroxide-based compounds. Furthermore, the relevant properties of the nanostructures and the implications on their performance for water treatment and contamination removal are highlighted. The hydrophilicity, pore size, skin thickness, porosity, and surface roughness of these nanostructures can help the water permeability of the nanoadsorptive membrane. Other properties such as surface charge modification and mechanical strength can improve the metal adsorption effectiveness of nanoadsorptive membranes during wastewater treatment. Various nanocomposite membrane fabrication techniques are also reviewed. This study is important because it gives important information on the roles of nanomaterials and nanostructures in heavy metal removal and wastewater treatment. Heavy metal discharge into wastewater from industry is a major environmental concern. The accumulation of these heavy metal contaminants in wastewater is particularly harmful to both humans and aquatic life; hence, the food chain process is negatively impacted ,2. Life However, scientific progress has expanded notice of water contaminant types and reawakened the necessity for comprehensive water treatment. Oxyanions (or oxoanions)  are formed by a number of redox-sensitive metalloids and metals, including titanium dioxide, iron oxides, manganese dioxide, aluminum oxides, and numerous oxide minerals ,15. Due In recent years, nanotechnology paired with known procedures has proved its usefulness in the water sector in eliminating pollutants and lowering toxicity and is emerging as a viable alternative to conventional wastewater treatment . NanomatCarbon-based nanomaterials, including carbon fibers, carbon nanotubes (CNT), activated carbons, graphene, graphene oxide, and other graphene-related materials are widely regarded as among the most effective adsorbents for extracting organic and inorganic contaminants from wastewater. Typically, these materials have a large specific surface area, strong chemical stability in acid/alkaline conditions, enhanced mechanical and thermal stability, and high porosity ,27. AlthMany research studies have shown that graphene oxide (GO) and reduced graphene oxide (rGO) have promising adsorption capabilities in the cleanup of heavy metal ions, organic pollutants, and pharmaceutical toxins ,32,33. GNanoparticles (NPs) derived from iron oxide are commonly employed as heterogeneous catalysts owing to their unique magnetic and physicochemical characteristics. Controlling the features of nanoparticles  and incorporating more nanoparticles into the system might increase the catalytic efficiency of iron oxide nanoparticles. It has been shown that iron oxide nanoparticles are effective adsorbents for removing heavy metals from water sources. For instance, nanohematite showed a good absorbent ability to remediate spiced tap water ionized metal ,37.2) nanoparticles. TiO2 nanoparticles showed high capability for Cu(II), Pb(II), Zn(II), Ni(II), and Cd(II) adsorption [Another type of metal-based nanoadsorbent is titanium dioxide , platinum (Pt), silver (Ag), and palladium (Pd) have high energies of ionization and as a consequence poor oxidation potential because of their short atomic size . Noble mHydrogels are three-dimensional, cross-linked, and elastic polymer networks with hydrophilic groups such as hydroxyl, amide, and carboxyl that expand in water and biological fluids ,43. Nume−1 for WU/W dry gel and 12.9 mg.g−1 for WU/W poly(amidoxime) in saltwater for six weeks [Using UV-polymerization, Jinxiang Gao et al. created a bio-inspired cellulose paper–poly(amidoxime) composite hydrogel. Due to its improved hydrophilicity, strong hydraulic/ionic conductivity, and broad-spectrum antibacterial activity, this hydrogel had a very efficient uranium capture capacity of up to 6.21 mg.gix weeks .Liu et al. reported that massive uranium recovery from seawater is a vital project to meet the long-term need for worldwide nuclear power, but it is still very difficult to produce high-efficiency adsorbents with good mechanical properties. In this study, a novel nanoclay–poly(amidoxime) (NC–PAO) double-network (DN) composite hydrogel adsorbent with high-efficiency, high strength, and rapid uranium extraction capacity was investigated .2, carbon nanotubes, Ag, etc.) for the elimination of organic/inorganic contaminants from wastewater [Nano-sponges are characterized by the presence of cavities and mesh-like/colloidal structures containing solid nanomaterials. They are suitable for capturing various materials ,48. Morestewater ,50. Nanostewater . Additiostewater ,52.Mohamed (2019) showed that a ternary α-Fe2O3/GO/WO3 nanocomposite enhances the photocatalytic degradation of crystal violet (CV) and methylene blue (MB) contaminants in water ,53.Additionally, the removal of fluoroquinolones  from wastewater was examined using a nickel ferrite nanocomposite modified with L-cysteine-linked 3-glycidyloxypropyltrimethoxysilane. The carboxyl groups of the L-cysteine molecule acted as the removing target moiety for fluoroquinolones .Magnetic oxide nanoparticles and hybrid carbon nanomaterials outperformed unmodified materials in terms of adsorption efficiency , which w2+ and Fe3+ [43− and Cr2O72−) and anionic organic dyes (such as methylene blue and Congo red). In another investigation, an Fe3O4/GO/LDH adsorbent was produced to remove Pd(II) and 2,4-dichlorophenoxyacetic acid from an aqueous system [3O4/LDH adsorbents [Layered double hydroxides (LDHs) are classed as anionic two-dimensional clays with distinctive layered structures and the chemical configuration of the inorganic layers exhibiting intriguing potential for dispersing many active sites ,59,60. Mand Fe3+ . They als system . In anotsorbents . SummaryWater is one of the most significant natural resources on earth, as it is essential for the survival of all living organisms and the development of humankind. In combination with industrialization and urbanization, water demand is expanding quickly, and scarcity has arisen as a serious financial constraint. Moreover, polluted water, namely, heavy metal pollution in water, has become a major environmental problem on a global scale. Electroplating, mining chemical plants, metallurgy, domestic wastewater, and agriculture can introduce heavy metals into water. Heavy metals such as zinc, lead, mercury, copper, and other elements may build up in the food chain and constitute a serious health danger to people. Heavy metals can, for example, harm kidneys, the central nervous system, the lungs, and other organs. Consequently, eliminating heavy metal ions from water is vital and has received much attention. Accordingly, nanomaterials have now been shown to be a viable option for eliminating heavy metals from wastewater . Heavy mRecently, nanotechnology has been incorporated with the majority of conventional methods to increase the rate of heavy metal removal from water bodies. In the last several decades, nanomaterials have attracted interest as a way to create cost-effective, ecologically acceptable, and extraordinarily effective methods for removing organic and inorganic toxins from the environment. Most nanomaterials have properties that make them ideal for water treatment or purification. Nanomaterials have a high sorption capacity and selectivity, allowing them to efficiently remove metal ions ,80.In recent years, carbon-based nanostructures including carbon nanotubes (CNTs) and graphene and its derivatives, such as graphene oxide (GO), have been utilized extensively for the separation of heavy metals. This is because of the non-toxic nature of these nanostructures as well as their increased adsorption capacity ,82. Thes2-hybridized graphite. When graphene is oxidized and exfoliated, graphene oxide (GO), a single-layer graphene with a high concentration of oxygen, is produced. GO’s usage in heavy metal adsorption has gained a great deal of attention due to its exceptional features, including an unusually high surface area and enormous amounts of oxygen-containing groups with considerable affinity for heavy metals, such as –COOH, –OH, and C=O groups. In addition, GO is known for its high hydrophilicity, which implies that it is very soluble in water [Due to their remarkable properties, graphene-based nanomaterials have become some of the most studied substances in recent years. Graphene is a hexagonally organized, single sheet of spin water .2+ and could be recycled at least four times [Following the collapse of a tailings dam in the Brazilian state of Minas Gerais, Lebron et al. used a modified version of the Hummers method to produce GO, and they tested the method’s capacity for removing heavy metals from a sample of natural surface water. The results revealed that Al, Cu, and Mn had the highest clearance rates of all metals, at 90.2%. Despite this, prolonged water consumption of aluminum would represent a moderate danger. The findings support GO’s ability to adsorb heavy metals, and its efficacy was confirmed even in intricate media . The porur times . Heavy mur times .Kadhim et al. modified the polyethersulfone (PES) membrane with graphene oxide nanoparticles (GO-NPs) to create mixed matrix membranes (MMMs) . This st2/g, making them an attractive choice for the adsorption-based removal of metal ions [In addition to mesopores, the diameter of CNTs spans from 1 to several nm, and their specific surface area is between 150 and 1500 mtal ions ,89. MWCNtal ions .Wang et al. synthesized a novel form of recyclable adsorbent by oxidizing an enhancer and modifying it with magnetic nanoparticles . In addi2+, Pb2+, Cd2+, and Sn2+, by employing a unique metal–carbon-based composite such ZnFe2O4-carbon nanotubes (CNT). The carbon nanotubes were ground with ZnFe2O4 to produce a black homogenized solid ZnFe2O4-CNT sorbent. The results indicated that the following parameters are optimal for efficiently recovering the investigated metal ions from aqueous solution at a dosage of 20 mg/100 mL with pH 5 for the Hg2+ and Pb2+ ions, pH 6 and an adsorbent dose of 50 mg for the Cd2+ and Sn2+ ions with a contact duration of 15 min, and a dosage of 20 mg/100 mL with pH 5 for the Hg2+ and Pb2+ ions, and the study indicated that linking CNTs with ZnFe2O4 improved heavy metal ion adsorption by 25 percent [Adam et al. (2021) created a simple and environmentally compatible material for eliminating heavy metal ions, including Hg percent .For the removal of heavy metal pollutants from wastewater, porous carbon compounds are regarded as valuable sorbents. These materials are typically mesoporous and macroporous, with limited micropore volume, and they facilitate the adsorption of heavy metals from the liquid phase. Because porous carbon materials are carbonaceous substances with a good degree of porosity and a large particle surface area, they have been a popular adsorbent for many years ,94,95. TMetal nanoparticles, such as silver and gold nanoparticles, have been used for years to remove heavy metal ions from wastewater. Silver, gold, and iron metal nanoparticles are the most extensively utilized adsorbent materials due to their huge surface area, outstanding optical and chemical properties, high absorptivity, and the ability to adjust their surfaces using various stabilizing and capping agents.2+, Hg2+, Cr2+, and so on [2+, Cd2+, and Pb2+ ions was 88.3%, 81.1%, and 72.6%, respectively. Furthermore, even after 4 months of storage, the walnut husk extract-produced silver nanoparticles remained stable, efficient, and thermally resistant. Overall, silver nanoparticles effectively removed heavy metals from petroleum effluent; however, the amount of heavy metal ions removed varied depending on the silver nanoparticle concentration, temperature, time, pH, and heavy metal ion dose [2+ and Pb2+ ion removal from aqueous solution using freshwater algal extract-synthesized silver nanoparticles. At an adsorbent dose of 0.5 g/L, the maximum adsorption capacity for lead was 23.98 mg/g, while the maximum adsorption capacity for cadmium was 22.47 mg/g. The higher the R2 number, the better the experimental data matches the Langmuir isotherm. The kinetics studies revealed that the Pb2+ adhesion to silver nanoparticle surfaces was due to the electrostatic attraction, whereas Cd2+ adhesion to silver nanoparticles surfaces was due to physical adsorption. According to rate kinetic calculations, the adsorption of Cd2+ and Pb2+ on silver nanoparticles followed pseudo-first-order kinetics, (R2 > 0.8) and pseudo-second-order kinetics (R2 > 0.9), respectively. This was determined by comparing the values of the rate constants. Finally, the findings suggest that silver nanoparticles might be applied as adsorbents in wastewater treatment to eliminate heavy metal ions in an effective manner [Based on many findings, metal nanoparticles may eliminate heavy metal ions from wastewater. There are several findings in the literature of silver nanoparticles interacting with pollutants including Cdnd so on ,98. In tnd so on ,100. Usind so on ,102. Itue manner .2+. To begin, the 15T bases were developed with the purpose of detecting Hg2+ by the coordination of T–Hg2+–T, which resulted in a detection limit of 9.6 × 10−9. Simply increasing the number of A bases just on one or either sides of the 15-mer sequences was all that was required to lengthen or shorten the 15-mer DNA sequence to generate the 25-mer and 59-mer sequences, respectively. The findings showed that decreasing the length of the DNA chain did not improve the detection sensitivity. Therefore, the acquired sensitivity towards the Hg2+ increased in the order of 15-mer < 25-mer < 59-mer aptamer. The apparent increase was attributed to the following factor: in the presence of Hg2+, T–Hg2+–T coordination ended up causing T base sequences to fall off the surfaces of the AuNPs, while additional A base sequences remained adsorbed on the surface of the AuNPs, which resulted in morphologically evident variations in grown AuNPs, allowing visible solution color changes [Piper auritum extract was employed as a reducing agent for AuNPs, and the composite was produced by ultrasonification. The ability of Mn2+ adsorption was tested using Dtm–AuNPs composites through varying the concentration of the composite from 1.0 mg/L−1 to 20 g.L−1. The adsorption procedure, which was aided by sonication, attained 98% adsorption effectiveness in 240 min. Furthermore, linear and non-linear pseudo-second-order models agreed well with the experimental data. Overall, the research demonstrated considerable Mn2+ adsorption using the Dtm–AuNPs composite, which is a novel, low-cost, and environmentally acceptable material for Mn2+ adsorption [Gold nanoparticles are commonly employed in the treatment of wastewater to detect and remove heavy metal ions . Gold na changes . Ruíz-Basorption .Verbascum thapus leaf extracts serving as reducing and stabilizing agents. When the generated nanoparticles were investigated for their capacity to reduce Cr(VI) ions, it was discovered that at a dosage of 1 g/L with a contact time of 30 min, they were able to completely eliminate the Cr(VI) [Iron nanoparticles have been extensively employed as adsorbents to eliminate heavy metals from wastewater due to their strong redox and adsorption abilities. It has been shown that the use of iron nanoparticles may effectively minimize the potential leachability of toxic substances via the process of in situ immobilization. In the beginning, materials made of iron were used in reactive subsurface permeability shields in order to extract chlorinated chemicals from groundwater. The development of nanotechnology led to the substitution of conventional ferromagnetic materials and large-scale iron particles with iron nanoparticles that have great activity. The findings of the experiments demonstrated that the reaction rate constant of iron nanoparticles was higher than that of conventional iron powder. Accordingly, it has been shown that iron nanoparticles may perform the functions of both reductants and catalysts in order to eliminate heavy metal pollutants . Recentle Cr(VI) . Hence, 2, MgO, ZnO, and CeO2 are extremely promising for the removal of heavy metal pollution from water bodies due to their large surface area and their ability to remove heavy metal ions, even at low concentrations.Nanoparticles of metal oxide have a high capacity for heavy metal ion adsorption. Nanoparticles of metal oxides such as CuO, TiO2+ and Fe3+ was evaluated by altering the physical parameters, including metal ions and adsorbent concentration, equilibrium contact duration, and aqueous solution pH. According to the findings, the adsorption capacity rose as the pH of the aqueous solution increased, with pH 6 being the optimal pH for adsorption of both metals. The kinetic analyses demonstrated that the adsorption of both metal ions on CuO NPs exhibited a pseudo-second-order behavior, and the equilibrium adsorption data suggested that the Langmuir isotherm model best represented Cd2+, while the Freundlich isotherm model best represented Fe3+. According to the findings, CuO NPs have the potential to act as an adsorbent in the process of removing heavy metal ions from the aqueous phase [2+, Ni2+, and Cd2+ ions was investigated. For Pb2+, Ni2+, and Cd2+, the optimum absorption capacity of each CuO NP was estimated as 88, 54, and 15 mg.g−1, respectively. Furthermore, Freundlich and pseudo-second-order models provided the best interpretation of the experimental results, and adsorption equilibrium was achieved in 1 h. These results suggest that CuO NPs could be a suitable nanoadsorbent for detoxifying water polluted with heavy metals [Copper is a naturally occurring and ubiquitous element in the environment. Its concentration in the Earth’s mantle is approximately 60 g/ton. Copper oxide nanoparticles, also known as CuO NPs, have been shown to be good adsorbent materials due to their low cost, ease of production, smaller size, greater surface area, abundant supply of their precursor material, and absence of toxicity. In wastewater treatment, CuO NPs are widely employed as an adsorbent to extract heavy metals, dyes, medicinal compounds, and other contaminants . The higus phase . Hassan y metals .2, among other metal oxide nanoparticles, have found widespread application for their ability to eliminate heavy metal ions. TiO2 nanoparticles are more promising than other metal oxide nanoparticles owing to their distinct physical–chemical characteristics  ions from aqueous solution. However, the metal ion concentration, amine type, and solution pH all had an influence on removal efficiency [2 nanoparticles are its complicated fabrication process and difficulty in separation when used in slurry suspension [TiOeristics , which iectively . TiO2 natal ions . Moreoveficiency ,117. Thespension .2+ ions from a copper chloride solution (10 ppm), while commercially available MgO had a removal capacity of 15% [2+ ions and Escherichia coli bacteria. The findings showed that 100 mg/L of MgO nanoparticles eliminated 50 mg/L Cd2+ ions while also inactivating 7-log bacterial cells. Furthermore, MgO nanoparticles were found to have increased bacterial inactivation in the presence of Cd2+ ions. This might be attributable to MgO’s direct contact with the bacterial cell membrane, which caused membrane degradation and the entry of Cd2+ ions into the cell body, resulting in intercellular component leakage. This research suggests that MgO nanoparticles might be a suitable material for wastewater treatment polluted by heavy metal ions and bacteria [3O4 nanocomposites. The adsorbent was found to have superparamagnetic characteristics with a surface area of 75 m2/g. According to the adsorption investigation, the maximum phosphate adsorption efficiency of 98.9% was attained after 20 min of contact time at 25 °C, pH 9, with a dosage of 5 g/L MgO/Fe3O4 nanocomposite and a metal ion concentration of 1 mg/L. This was accomplished using a concentration of metal ions and a dosage of 5 g/L MgO/Fe3O4 nanocomposite. The adsorption process was exothermic and spontaneous, as evidenced by the thermodynamic characteristics, and the pseudo-second-order kinetic model suited the experimental results well [3+, Cu2+, and Zn2+ ions. The maximal absorption values were found to be 41.36, 13.76, and 7.23 ions/nm2 for Cr3+, Cu2+, and Zn2+, respectively. Furthermore, when the pH of the metal ions was elevated from 7 to 11, the adsorption rate increased. These findings suggest the MgO- and NiO-embedded silica-based nanocomposite may be a candidate material for eliminating heavy metal pollutants from water bodies [As adsorbents for heavy metal ions, magnesium oxide offers several benefits, which include an exceptional adsorption capacity, low cost, low toxicity, good availability, and an eco-friendly behavior . Metal iy of 15% . Cai et bacteria . MgO nanlts well . Abuhatar bodies .6+, Ag+, Cu2+, Mn2+, Pb2+, Ni2+, and Cd2+ was investigated by Le et al. It was shown that the ZnO nanoparticles generated via solid precipitation had a rod-like structure, with an average length of 497 nm and a diameter of 75 nm, respectively. According to the findings, ZnO nanoparticles were very effective in removing Cu2+, Ag+, and Pb2+ ions, with an efficiency of removal that was more than 85%. However, the removal effectiveness of Cr6+, Mn2+, Cd2+, and Ni2+ ions was shown to be less than 15%. In addition, it has been stated that the process for heavy metal ion removal was found to be reduction for Cu2+, Ag+, and Cr6+ ions; oxidation for Pb2+ and Mn2+ ions; and adsorption under UV irradiation for Cd2+ and Ni2+ ions. All of these pathways were discovered to be responsible for heavy metal ion separation or removal [2/g, these nanoparticles were put through a series of tests to see how well they removed Cr3+ metal ions. In adsorption experiments, ZnO nanoparticles were shown to have a greater affinity for Cr3+ ions, with a maximum adsorption of 88.5 mg/g for 1 g of adsorbent in 1 L of water. Moreover, the pseudo-second-order and Langmuir isotherm models best describe the adsorption experiment data. As a direct consequence of this, the ZnO nanoparticles that were generated have the potential to be useful material for the filtration of dental effluent [ZnO nanostructures possess distinguished physicochemical and biological properties that make them distinctive from ZnO in bulk form . The key removal . Gu et aeffluent .2 nanoparticles have also been studied in terms of specific surface area, indicating that nanoparticles are potent adsorbents with size-dependent properties [2 nanoparticles as a function of the starting concentrations of both metal ions and nanoparticles as well as pH was created. The study discovered that CeO2 nanoparticles had the highest absorption for Cd2+ at 93 mg/g, Pb2+ at 128 mg/g, and Cr6+ at 34.4 mg/g. Furthermore, the findings showed that the sorption capacity of CeO2 nanoparticles was approximately equivalent in both single and multiple systems, and that the initial metal ion and nanoparticle concentrations were significantly predictive of all three metals’ sorption capacities. The sorption capacity of cadmium and chromium was shown to be affected by pH but not lead. This research paves the way for the novel use of CeO2 nanoparticles in the capacity of an adsorbent for complex waterways that are contaminated with heavy metals. In situ growth and self-assembly were employed to assess the capacity of CeO2 nanoparticles to remove Cd2+, Pb2+, and Ce6+ from an aqueous solution when combined with reduced graphene oxide. The in situ-grown CeO2/rGO nanocomposite had the highest lead adsorption rate (95.75 mg/g), whereas the self-assembled nanomaterial had the highest cadmium adsorption rate (31.26 mg/g), according to the data. The kinetic experiments found that the Langmuir model matched the lead removal best, whereas the Freundlich and Langmuir models fit the cadmium removal best. Although CeO2/rGO showed strong Cd2+ and Pb2+ adsorption capacities, only a minor concentration of Cr6+ was removed [2/rGO nanocomposite produced via in situ growth could be a potential material for wastewater treatment.Cerium oxide is a non-toxic rare-earth metal oxide that has found widespread application in a variety of fields, including photocatalysis, UV blocking, sensor technology, and the treatment of wastewater . CeO2 naoperties . Contrer removed . Accordi3O4/polydopamine-grafted L-cysteine (Fe3O4/PDA/L-Cys) core shell composite and investigated its efficiency in removing Pb2+ ions from aqueous solution. At pH ranges between 2.8 and 6.0, the Pb2+ removal effectiveness of Fe3O4/PDA/L-Cys was evaluated. The research demonstrated that adsorption increased as pH rose. At a concentration of 1.5 g/L, the maximum Pb2+ ion adsorption was reported at a pH of 6. The kinetic analyses revealed that the adsorption conformed well to the pseudo-second-order and that the Fe3O4/PDA/L-Cys removal was spontaneous and endothermic [2O3/CeO2/Fe3O4 nanofibers and studied their efficiency in removing fluoride ions. The produced La2O3/CeO2/Fe3O4 nanofibers demonstrated good defluoridation, with a maximum absorption of 229 mg/g recorded. The findings of thermodynamic analysis demonstrated that the defluoridation process was spontaneous and endothermic. Kinetic investigations demonstrated that the adsorption mechanism was well-described by a pseudo-second-order model. The prepared La2O3/CeO2/Fe3O4 nanofibers were good at regenerating, which suggests that La2O3/CeO2/Fe3O4 nanofibers are stable and promising options for defluoridating wastewater [The application of iron oxide nanoparticles for the elimination of toxic metals from wastewater is more attractive due to their unique properties, such as their magnetic properties, huge surface area, and tiny size. Because iron oxide nanoparticles are magnetic, it is simple to extract adsorbent materials from the solution for easy reusability . Yang etothermic . In anotothermic . Likewisstewater .Saccharum ravannae, Saccharum officinarum, and Oryza sativa leaf extracts. The ability of biomass-derived silica nanoparticles to remove Cu2+ and Pb2+ from wastewater was investigated further. In addition, the Freundlich isotherm model and pseudo-second-order kinetics were employed to characterize the adsorption process [Materials made from biomass are widely available, cheap, and have very little to no economic value. Despite all of these factors, they are more prevalent and pose several handling and disposal challenges . Multipu process . Chen et process . NanomatA wide range of processes, such as reverse osmosis, coagulation, membrane filtration, chemical reduction, electrolytic removal, ion exchange, flocculation, precipitation, phytoremediation, photocatalytic degradation, electrodialysis, and adsorption, have been utilized in the treatment of wastewater . Due to In the 1980s, membrane adsorbent technology was created by combining membranes with adsorbents that bind molecules through physical or chemical interactions . BroadlySarah Glass et al. developed the post-modification of polyacrylonitrile (PAN) membranes with primary, secondary, tertiary, and quaternary amines using different modification reactions. In comparison to other membrane materials, PAN is chemically easier to modify, since it is carrying a nitrile group. Several modification reactions of nitrile-carrying substances such as PAN are known. Nevertheless, these modification reactions were usually not applied on PAN ultrafiltration (UF) membranes, which require heterogeneous reactions in order to preserve the membrane structure .Adsorbents such as carbon-based nanomaterials, zeolites, polymers, agro-waste, ceramics, metal oxides, composites, and hybrid materials have been widely used for the adsorptive removal of a wide range of pollutants from contaminated water. This procedure is referred to as adsorption. Adsorption refers to the physical and chemical interaction that happens between an adsorbate and an adsorbent. The adsorption process is affected by a variety of characteristics, including but not limited to temperature, medium pH, adsorbent–adsorbate interaction forces, concentration, the presence of foreign components, and others ,161.The adsorption process is based on mass transfer in which a material accumulates at the interfaces of two phases, such as the contact between liquid and solid, liquid and liquid, or gas and solid . This prSome nanoparticles can selectively remove a large variety of specific substances from polluted water . When coIt has been found that the mechanical strength of adsorbent nanocomposite membranes is an outward sign of their durability . ResearcAccording to Mukherjee, the mechanical properties of the polysulfone/graphene oxide nanocomposite membrane rose as the graphene oxide concentration rose from 0 to 0.1 percent by weight in the polysulfone membrane. The formation of a connection between two polysulfone polymer chains by graphene oxide nanoparticles resulted in greater contact between the nanocomposite membranes. However, when the graphene oxide nanoparticle loading climbed to 0.5% weight, the membrane’s mechanical strength decreased before the pore density in the membrane structure increased .2/Al2O3/SiO2 PVDF nanocomposite membrane) in tensile strength between control and nanocomposite membranes, the elongation at break for composite membranes decreased dramatically [Amir Razmjou et al. found that with minor differences , Cr (VI), and Cu (II) by 205.7 mg/g, 175.67 mg/g, and 183.38 mg/g respectively, at a pH of 7, 2, and 3, respectively .2 [Graphene oxides, on the other hand, are utilized to remove heavy metals by increasing the adsorption behavior, since they can be readily controlled by surface functionalization using various functional groups . The eli2 .In another research study, nickel iron oxide was added to polysulfone nanocomposite membranes, at which point the membrane’s zetapotential plummeted from 8.8 mV to −49 mV. In order to attain an equilibrium, hydrophilic NFO nanoparticles were distributed over the polymer–water interface during phase inversion. This resulted in negative surface charges of the adsorbent membranes .2, and –SO3H groups. These functional groups are needed for the attachment of metal ions; hence, they are eliminated upon contacting the membrane surface, regardless of whether the pore diameters are larger than the metal ion size. To properly define the removal mechanism of heavy metals via adsorptive membranes, it is essential to recognize that their surfaces are charged by ionization, and that the adsorbent may include charged ionic functional groups. For the removal of arsenic from water, Rowley et al. created polyethersulfone (PES) nanocomposite membranes utilizing surface-modified Fe3O4 nanoparticles (NPs). The PES membranes produced with A–Fe3O4 NPs exhibited effective adsorption capabilities with low concentrations of NPs , indicating a promising impact for industrial applications in the water treatment sectors. PES membranes with 3 wt.% A–Fe3O4 NPs had the greatest rejection rate of 76% and the highest observed arsenic equilibrium adsorption capacity of 14.6 mg/g [Through ion exchange or surface complexation, adsorptive membranes can be changed to incorporate reactive functional groups such as –COOH, –NH4.6 mg/g .In this review, we discuss different techniques that are employed to fabricate the different kinds of nanocomposite membranes. Firstly, the nanocomposite membranes are differentiated into four types: conventional nanocomposites, thin-film nanocomposites, thin-film composites with nanocomposite substrates, and surface-located nanocomposites and mixed matrix nanocomposite membranes (MMNMs) . MMNMs aUsually there are three approaches for the first step (homogenous solution synthesis of nanocomposite membranes). The polymer is added to the solvent initially, and then the nanofiller is added to the mixture in one method. In another, the nanofiller is added to solvent and the polymer to that mixture. In a third method, the polymer and nanomaterials are dissolved in solvent separately and combined to yield a homogenous mixture . Of thesThen the different fabrication techniques come into play in producing nanocomposite membranes.Phase inversion is the controlled de-mixing (liquid–liquid de-mixing) of homogeneous polymer solutions into two solid and liquid phases . In the Four types of nanofillers employed in the phase inversion procedure are as follows: (1) organic material; (2) inorganic material; (3) hybrid material containing two or more types of material; and (4) biomaterial. Hydrophilicity, permeability, porosity, pore interconnectivity, and antifouling abilities are all improved by utilizing diverse combinations of polymers and nanofillers.Polyanaline nanofibers were utilized as a nanofiller in the manufacture of nanocomposite membranes, increasing attributes such as membrane strength as well as hydrophilicity and pore size, which enhanced permeability through the polymer ,185.In one study, the combination of inorganic metals such as silver with polysulfone increased the membrane’s pore size, hydrophilicity, and antibacterial characteristics .3O4 nanoparticles were allowed to dissolve in N,N-dimethylacetamide to create a nanocomposite membrane (400 rpm for 24 h). Polyaniline (PANI)/Fe3O4 nanoparticles were initially dispersed in N,N-dimethylacetamide using ultrasonication before being added to a polyethersulfone solution due to their magnetic nature [Using a phase inversion method, specific proportions of polyethersulfone and polyaniline (PANI)/Fec nature .The instantaneous phase inversion between the nonsolvent and solvent in the coagulation bath results in the creation of a permeable membrane with macrovoid gaps, a thin top layer, and finger-like cavities. This is due to the vast quantities of heavy metal ions that flow through the membrane. As per the results of a number of research studies, this technique increases the number and size of pores, which consequently enhances the diffusion rate of nonsolvents ,189.The vast majority of microporous membranes that are used in microfiltration and ultrafiltration are created by the processes of solution casting (or an extraction method), dry stretching, and particle stretching ,191. In A porous membrane is produced by a variety of procedures. In the first phase of the process, a precursor film with a row-nucleated lamellar structure is produced by shear- and elongation-induced crystallization of the polymer. This polymer is chosen for its high molecular weight and even distribution of molecular weight. The second stage involves annealing the precursor film at temperatures close to the melting point of the resin. This will assist in eliminating crystalline phase defects and increasing lamellae thickness. Finally, the pores are extended at low and high temperatures in order to develop and expand them suitably .In one study, nanostructured stretched polytetrafluoroethylene (PTFE)–sulfonated polystyrene (PS) ion-exchange membranes were investigated for their suitable physical and chemical characteristics as well as their efficiency. To implant the PS in the stretched PTFE film, high-temperature polymerization of styrene, which was absorbed at 90 degrees Celsius from a solution consisting of styrene, divinylbenzene, and toluene, was utilized. According to the findings of the researchers, the membranes that were produced had a permeability and diffusion coefficient for methanol 1.5 times lower than those for water .In addition, the initial stage in the creation of high-density polyethylene (HDPE) hollow fiber membranes was conducted to stretch the precursors of melt-spun hollow fibers. Consequently, technologies for producing slit-shaped holes by cold and hot stretching have been developed effectively. It was observed that the take-up rate affected both the microporous structure and the crystalline structure of the precursors of the stretched membrane .Electrospinning is a unique fabrication method and a straightforward nanofiber production technology that may be used to generate nanofibrous nonwovens. It is a basic and simple process for producing nanofibers with the potential for cost-effective development on laboratory, pilot, and industrial scales. Moreover, electrospun nanofibrous materials offer distinct benefits when used as membranes due to their specific properties, which include small fiber diameters, high porosity, large specific surface area, flexibility in surface functionality, excellent pore size adjustability, and pore interconnectivity . These aElectrospinning is gaining popularity as a versatile and trustworthy technology for producing nanofibers for application in porous membrane production, such as water filtration and desalination. Electrospun membranes were used to separate polystyrene particles of 1, 5, and 10 m diameters. Electrospun membranes successfully removed more than 90 percent of the microparticles from solution. This study sets the way for future research on the use of nanofibers as screens in separation techniques for water pre-treatment prior to reverse osmosis or as pre-filters to reduce fouling and pollution before ultra or nanofiltration .In recent years, significant advancements have been achieved in relation to this topic, and the technology in question has been used in a diverse array of applications. To better regulate nanofiber shape, structure, surface functionality, and assembly techniques, researchers have concentrated on either better understanding of the fundamental principles of electrospinning or identifying optimum electrospinning settings for diverse polymers and biopolymers ,198,199.Nanoparticles or nanofillers included in the spinning fluid can enhance the properties of electrospun nanocomposite membranes . One stu2) particles were effectively embedded in nylon-6 spider-net-like nanofiber mats. The antifouling effect, mechanical strength, antibacterial protection, and UV protection were all enhanced by the introduction of a small amount of TiO2 NPs to the electrospun mats [In one study, tiny titanium dioxide . Using nylon 6/silver as a barrier against mposites .2/h at 0.4 bar working pressure [Hosseini et al. produced and employed montmorillonite (Mt) chitosan/poly (PVA) nanocomposite electrospun nanofibrous membranes (ENM) to treat colored wastewater. Mt with varied mass percentages  was included in the membrane structure, and its influence on the pore size, morphology, porosity, mechanical strength, and permeation capabilities of ENM was examined. The membranes were employed as affinity screens for dye removal through ultrafast penetrating adsorption. Adding Mt as a strengthening element increased the nanocomposite ENM’s resistance to compaction, as demonstrated by the findings. Young’s modulus rose from 0.89 MPa for chitosan/PVA ENM to 2.09 MPa for nanocomposite membranes containing 1% Mt. It was discovered that the optimal concentration of Mt for increasing both water flow and dye removal was 2.0 mass percent. Effectively resolving the trade-off between permeability and selectivity, the improved membrane repelled 95 percent of Basic Blue 41 (BB41) at a flux of 1765 L/mpressure .Despite the advances made in the use of nanotechnology-based approaches for water treatment, several obstacles and challenges still need research efforts and considerations to realize the promise of these technologies. A major limitation is the fouling of the nanoadsorptive membrane through agglomeration of extraneous particles in the membrane. This problem can be solved through the introduction of nanoparticles with anti-fouling characteristics, which can be produced by the processing, functionalization, and surface modification of these nanomaterials. Additionally, further approaches can be pursued to avoid the regrowth of microbes on the surface of membranes during water treatment processes. The comparatively poor adsorption capacity of the adsorptive systems is an additional issue requiring more investigation. The membranes’ adsorption capacity is affected by the limited number of adsorbents that can be included into the membrane matrix. To determine the practicality of deploying adsorptive membranes on an industrial level, exhaustive cost–benefit evaluations should be done. Moreover, the technological requirements of using adsorptive membranes on the industrial scale still need further investigation.In addition, the safety and nontoxicity of the adsorbent materials introduced to the membrane is a critical problem to be solved. Toxic fillers are used in water purification, resulting in contaminated water. There should be no danger to individuals or to the environment associated with any water treatment method used. Multiple tests and experiments can be used to determine the quality of water generated—whether it is acceptable for human consumption or for discharge into the aquatic environment. Moreover, many of the published articles highlighted the effectiveness of the materials and adsorbent systems in removing heavy metals and other pollutants from water and the environment, but they do not sufficiently address the reusability of the contaminants that have been removed from the environment. The idea of reusing the heavy metals and other pollutants removed from industry waste streams should be investigated further.Furthermore, the development of novel nanomaterials with better functions for adsorptive membranes warrants further research. Even with the evaluation of many materials in the laboratory, there are still drawbacks in finding new materials that could be used in mixed matrix membranes for better adsorption of heavy metals. The high cost of producing these materials is also a significant factor to consider. The possibility of finding and developing novel nanomaterials with affordable cost and superior characteristics that can break into the market for the commercial scale-up of adsorptive membranes should also be researched. Finally, the future outlook in developing nanotechnology-based water treatment solutions is promising, and the advances in the application of these technologies are touted to deliver reliable answers to water treatment and environmental cleanup issues.In summary, the use of nanotechnology in many contexts has led to advancements in engineering and environmental research, as well as to the development of new nanotechnology-based technologies. Extremely harmful to individuals and the environment are the high levels of undesired or unrecognized pollutants that make wastewater treatment a global priority. Emerging nanoscale technology has driven the advanced use of new and low-cost techniques, such as nanoadsorptive membrane processes that are suitable for filtration and adsorption procedures for the removal of environmental toxins. In recent years, improvements in membrane qualities such as thermal properties, selectivity, hydrophilicity, and permeability have been made. This can be advanced by the production of more low-cost and efficient polymeric nanocomposite membranes. In addition, it has been established that the incorporation of nanoparticles into the polymer matrix improves the chemical, mechanical, and thermal characteristics. This review focuses on the introduction of several nanomaterials into membranes in diverse water treatment applications.


Cholangiocarcinoma (CCA) is the malignancy originating from the biliary epithelium, including intrahepatic (iCCA), perihilar (pCCA), and distal (dCCA) CCA. The prognosis of CCA is very poor, and the biomarkers of different CCA subsets should be investigated separately. Holliday junction recognition protein (HJURP) is a key component of the pre-nucleosomal complex, which is responsible for normal mitosis. The ectopic expression of HJURP has been reported in several cancers, but not CCA.In our study, we investigated the expression of HJURP in 127 CCA patients which were composed of 32 iCCAs, 71 pCCAs, and 24 dCCAs with immunohistochemistry and divided these patients into subgroups with a low or high expression of HJURP. With chi-square test and univariate and multivariate analyses, we evaluated the clinical relevance and prognostic significance of HJURP in iCCAs, pCCAs, and dCCAs.HJURP was ectopically upregulated in CCAs compared with the para-tumor tissues based on TCGA and other mRNA-seq databases. A high expression of HJURP was correlated with low overall survival rates of iCCA and pCCA, but not in dCCA. Moreover, HJURP was an independent prognostic biomarker in both iCCA and pCCA. Patients with high HJURP were more likely to suffer CCA-related death after operation.HJURP was an independent prognostic biomarker in both iCCA and pCCA, but not in dCCA. Our results provide more evidence of the molecular features of different CCA subsets and suggest that patients with high HJURP are more high-risk, which can guide more precision follow-up and treatment of CCA. Cholangiocarcinoma (CCA) is the malignancy originating from the biliary epithelium and has molecular features of cholangiocyte differentiation , 2. CCA via its highly conserved Scm3 domain  is a unique histone H3 variant responsible for organizing functional centromeres , 9. As a3 domain –13. The 3 domain . As for 3 domain –16. As a3 domain –19. HoweIn our study, we investigated the expression of HJURP in 127 CCA patients comprising 32 iCCAs, 71 pCCAs, and 24 dCCAs with immunohistochemistry (IHC) and divided these patients into subgroups with a low or high expression of HJURP. With chi-square test and univariate and multivariate analyses, we evaluated the clinical correlation and prognostic significance of HJURP in iCCAs, pCCAs, and dCCAs.Our cohort was composed of 127 consecutive CCA patients who underwent radical surgery and received regular follow-up. The cohort consisted of 67 male patients and 60 female patients. All the specimens were obtained with the prior consent of patients, and the study was approved with the Ethics Committee of Qilu Hospital of Shandong University and the Second Hospital Affiliated to Shandong First Medical University. The tumors were classified and staged according to the 8th edition of the AJCC/UICC TNM classification system.Tissue microarray (TMA) was used for IHC detection. Hematoxylin and eosin (HE) staining was performed to confirm the diagnosis and design the expected area for TMA histological features of all samples. Core biopsies 1.0 mm in diameter were taken from each sample and arranged into TMA slides.For IHC, TMA slides were first de-paraffined and rehydrated and then incubated in sodium citrate buffer (pH = 6.0), which was boiled by a microwave for optimal antigen retrieval. After that, TMA slides were immersed in hydrogen peroxide for 15 min to minimize the endogenous peroxidase activity. After incubation in 5% bovine serum albumin for 30 min to inhibit the unspecific binding, the TMA was incubated in a primary antibody  at 1:50 overnight at 4°C and then in a secondary antibody  at room temperature for 1 h. DAB was finally applied for the visualization of antigen.The tumor area was selected by a senior pathologist, and the IHC results of the tumor area were semi-qualified by the IHC score, which was the product of stained cells and staining intensity. The stained cells were classified as <25%, 25%–50%, 50%–75%, and ≥75%, representing scores 1, 2, 3, and 4, respectively. The staining intensity was stratified as negative, weak, moderate, and strong staining, representing scores 0, 1, 2, and 3, respectively. Therefore, the final IHC score was set from 0 to 12. The cutoff was defined with the receiver operating characteristic (ROC) curve and applied to divide the cohort into high- or low-HJURP subgroup.P value < 0.05 was considered statistically significant.Statistical significance was analyzed with software SPSS 22.0. The correlation between HJURP and clinicopathological factors was assessed by the chi-square test. The Kaplan–Meier method was applied to plot the survival curves, and the log-rank test was administrated to generate the statistical significance of different subgroups. The independent prognostic significance of HJURP and other clinicopathological factors was estimated with multivariate analysis by the Cox proportional hazards regression model. From 2014 December to 2019 May, a retrospective CCA cohort was established, consisting of 127 CCA patients who underwent radical surgery. This cohort was composed of 32 iCCAs, 71 pCCAs, and 24 dCCAs (P = 0.114) and histological grade (P = 0.059) in iCCA, tumor size (P = 0.076), and N stage (P = 0.089) in pCCA as well as tumor size in dCCA (P = 0.058) approached to be statistically significant.Several clinicopathological factors including the gender and age of patients, tumor size, histological grade, tumor infiltration (T stage), lymphatic invasion (N stage), and TNM stage were enrolled to investigate the potential HJURP-associated factors in CCA (P = 0.184), which may be a result of that the sample size of the dCCA cohort was not that large (n = 24) and pCCA . The odds of CCA-related death for patients with high HJURP was 5.23- and 3.25-fold higher than those for patients with low HJURP. In dCCA, the independent prognostic significance of HJURP was not notable (P = 0.157). In addition, large tumor size (P = 0.012) and advanced T stage (P = 0.033) were also defined as independent factors suggesting poor outcome in iCCA. In pCCA and dCCA, no other clinicopathological factors were identified as independent prognostic factors in the Cox-regression hazard model.All the above clinicopathological factors were selected into the Cox-regression hazard model for multivariate analysis (The centromere is a conserved eukaryotic protein machinery required in precision segregation of the parental genome into two daughter cells during mitosis. HJURP is a key member of the CENP-A pre-nucleosomal complex, essential for the deposition of CENP-A at the centromeres. Accordingly, HJURP should be mainly expressed in the nucleus; however, we mainly observed HJURP in cell cytoplasm. We also noticed this inconsistency in HJURP sub-localization in previous studies. In hepatocellular carcinoma, prostate cancer, ovarian cancer, and breast cancer, IHC showed that HJURP was mainly expressed in cell cytoplasm , 21–24; Emerging molecular differences between iCCA, pCCA, and dCCA were revealed, thanks to the new high-throughput techniques such as whole-genome sequencing and transcriptome sequencing. There are several molecular classifications of CCA, especially iCCA, based on the genetic alterations , 26. HowCCA is featured with the poor prognosis and limited treatment options . To idenIn this study, we did not further investigate the molecular mechanism of HJURP-involved CCA prognosis and the role of HJURP in CCA progression. The molecular mechanism of how HJURP participated in cancer progression had various explanations. Previous studied reported that several key proteins and signaling pathways were involved in HJURP-associated cancer progression. The involved proteins include CDKN1A, SPHK1, p21, and p53, and the signaling pathways contain GSK3β/JNK signaling, MAPK/ERK1/2 signaling, AKT/GSK3β signaling pathways, and so on , 21, 22.In summary, we detected the expression of HJURP in 127 CCA patients and demonstrated that HJURP was an independent prognostic biomarker in both iCCA and pCCA, but not in dCCA. Our results can provide more evidence of the molecular characters of different CCA subsets and suggest that patients with high HJURP are high-risk, which can guide more precision follow-up and treatment of CCA.Publicly available datasets were analyzed in this study. This data can be found here: GSE33327,GSE132305.This study was reviewed and approved by the study was approved with the Ethics Committee of Qilu Hospital of Shandong University and the Second Hospital Affiliated to Shandong First Medical University. The patients/participants provided their written informed consent to participate in this study.YY, JY, ZL, and WC performed the experiments, YY and PL collected the specimens and analyzed the data, and PL wrote the paper. All authors contributed to the article and approved the submitted version.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.


The purpose of this study is to evaluate the optimal dose of tirzepatide (TZP) for the treatment of type 2 diabetes mellitus (T2DM) by meta-analysis and trial sequential analysis (TSA).Clinical trials of TZP for T2DM were obtained by searching 8 databases with a time limit from database creation to May 2022. Mean differences (MD) and 95% confidence intervals (95%CI) were used for continuous variables, and relative risk (RR) and 95%CI were used for dichotomous variables.P < 0.00001), fasting serum glucose (FSG)  and weight , and TZP 15 mg significantly reduced HbA1c , FSG  and weight . Compared with TZP 10 mg, TZP 15 mg dramatically reduced HbA1c , FSG  and weight . The TSA indicated that the benefits observed in the current information set were conclusive, except for the FSG of “TZP 15 mg vs. TZP 10 mg”. In terms of safety endpoints, meta-analysis revealed that there was no significant difference in the serious adverse events (AEs), major adverse cardiovascular events-4 (MACE-4), cardiovascular death, hypertension, cancer and hypoglycemic of the three dose groups of TZP. Compared with TZP 5 mg, TZP 10 mg increased total adverse events  and gastrointestinal (GI) AEs , and TZP 15 mg increased total AEs . There were no significant differences in total AEs and GI AEs for TZP 15 mg compared to TZP 10 mg. The TSA demonstrated that the total AEs of “TZP 15 mg vs. TZP 5 mg” were conclusive.Compared with TZP 5 mg, meta-analysis showed that TZP 10 mg significantly reduced glycosylated hemoglobin type A1c (HbA1c) (MD −0.24, 95%CI −0.31~-0.17, TZP 15 mg >TZP 10 mg > TZP 5 mg in terms of lowering glycemia and reducing weight. TZP 5 mg > TZP 10 mg = TZP 15 mg in terms of safety. On this basis, we recommend TZP 5 mg as the first-choice dose for patients with T2DM to minimize AEs while reducing glycemia and weight. If patients cannot effectively control their glycemia after taking TZP 5 mg, it is recommended to take TZP 15 mg directly to achieve the best effect of glycemic reduction. However, most of the included studies have the background of basic medication, the independent efficacy and safety of different doses of TZP still need to be tested.Unique Identifier: CRD42022341966. Type 2 diabetes mellitus (T2DM) is a chronic progressive metabolic disease characterized by pancreatic β-cell dysfunction as well as insulin resistance . AccordiThe latest study indicates that tirzepatide (TZP) has a promising application as a novel hypoglycemic drug in the treatment of T2DM . TZP is This study strictly followed the systematic review and meta-analysis methodology of the Preferred Reporting Items for Systematic reviews and Meta-Analyses (PRISMA) .The databases of China National Knowledge Infrastructure (CNKI), China Biology Medicine (CBM), VIP, Wanfang, Embase, PubMed, the Cochrane Library, and Web of Science were searched for clinical studies of TZP for T2DM, all with a time limit from database creation to May 2022. English subject terms included tirzepatide, type 2 diabetes mellitus, and Chinese subject terms included tirzepatide, erxing tangniaobing (the Chinese name of type 2 diabetes mellitus). On the basis of the subject terms, the Chinese free words were expanded with CNKI and CBM databases, and the English free words were expanded with MeSH database. The subject terms and free terms were then combined for the search.The inclusion criteria are shown below. (1) Study design: Randomized controlled trial. (2) Participants: The basic diagnosis of type 2 diabetes mellitus was met . (3) IntThe exclusion criteria were as follows. (1) Studies such as reviews, animal experiments, and case reports. (2) Studies published repeatedly. (3) Studies published in abstract form. (4) Studies with incomplete or unclear data. .In the initial step, the base literature retrieved from each database was imported into Endnote X9, then duplicates were eliminated in turn, and the literature was eliminated based on the inclusion criteria after reading the title and abstract, and reviewing the full text to finalize the included literature. In the next step, the included literature was categorized and organized to extract basic characteristics such as authors, year, sample size, mean age, sex ratio, intervention, and duration of treatment, which were entered into a statistical table of information. In the final step, the risk of bias was assessed in accordance with the requested entries by using the Cochrane Risk of Bias Assessment Tool. All the above work was carried out independently by two investigators, and any disagreement was decided by a third investigator.I2 test and Q test. If I2 < 50% and P > 0.1, the heterogeneity was small and fixed-effects model analysis was carried out. Otherwise, random effects model analysis was used. Sensitivity analysis was used for indicators with significant heterogeneity, whereby the remaining studies were combined for analysis after excluding one study at a time. If there was no noticeable change in the continuous variables obtained from each combined analysis, the results were suggested to be robust.Meta-analysis was conducted using Revman5.3. And relative risk (RR) and 95% confidence intervals (95% CI) were employed as effect sizes for dichotomous variables. Mean differences (MD) and 95% CI were utilized as effect sizes for continuous type variables. Heterogeneity was performed by P > 0.1, there was no publication bias. GRADEpro3.6 software was utilized to evaluate the quality of evidence, and the method of evaluation was based on the GRADE evidence evaluation guidelines.TSA0.9.5.10Beta software was applied to perform the TSA. If the cumulative Z value crossed the required information size or TSA threshold, the original result was conclusive. Publication bias was evaluated using Stata15.0 software. If the funnel plot revealed that the scatter on both sides is essentially symmetric and the Harbord regression displayed A total of 312 studies were obtained from the search. The 156 duplicates were eliminated. One hundred and twenty eight studies were eliminated after reading the titles and abstracts. Twenty two articles studies were excluded after reading the full text. The reasons for full-text rejection were as follows: those published in abstract form, those published with duplicate data, and those with inconsistent outcome indicators. Six studies were finally included –29. As sA total of six clinical studies were included –29 with In the six included studies, Del 2021  and LudvP < 0.00001), and TSA showed that the cumulative Z value crossed the required information size in the second study (RIS = 529), which revealed that this benefit observed for the current information set were conclusive. Compared with TZP 5 mg, meta-analysis demonstrated that the TZP 15 mg could significantly reduce HbA1c by 0.37% , and TSA showed that the cumulative Z value crossed the required information size in the second study (RIS = 223), which revealed that this benefit observed for the current information set were conclusive. Compared with the TZP 10 mg, meta-analysis demonstrated that the TZP 15 mg was able to significantly reduce HbA1c by 0.12% , and TSA showed that the cumulative Z value crossed the required information size in the fourth study , which revealed that this benefit observed for the current information set were conclusive. The GRADE evaluation showed there were no serious risk of bias, inconsistency, indirectness and imprecision, and the quality of evidence for each of these indicators was high. As shown in Compared with TZP 5 mg, meta-analysis demonstrated that the TZP 10 mg could significantly reduce HbA1c by 0.24% , and TSA showed that the cumulative Z value crossed the required information size in the third study , which suggested that the benefits observed in the current information set were conclusive. Compared with TZP 5 mg, meta-analysis revealed that TZP 15 mg significantly reduced FSG by 8.52 mg/dl , and TSA showed that the cumulative Z value crossed the required information size in the second study (RIS = 525), which suggested that the benefits observed in the current information set were conclusive. Compared with TZP 10 mg, meta-analysis revealed that TZP 15 mg significantly reduced FSG by 2.73 mg/dl , and TSA showed that the cumulative Z value had not reached the required information size  or TSA boundary value, which suggested the benefits needed to be confirmed by more research. The GRADE evaluation displayed high quality of evidence for all indicators except for the FSG of “TZP 15 mg vs. TZP 10 mg”, which had moderate quality of evidence. As shown in Compared with TZP 5 mg, meta-analysis revealed that TZP 10 mg significantly reduced FSG by 5.82 mg/dl , and TSA showed that the cumulative Z value crossed the required information size in the second study (RIS = 447), which suggested that the benefits observed in the current information set were conclusive. Compared with TZP 5 mg, meta-analysis showed that TZP 15 mg significantly reduced weight by 4.63 kg , and TSA showed that the cumulative Z value crossed the TSA boundary value in the first study (RIS = 185), which suggested that the benefits observed in the current information set were conclusive. Compared with TZP 10 mg, meta-analysis showed that TZP 15 mg significantly reduced weight by 2.18 kg , and TSA showed that the cumulative Z value crosses the required information size in the second study (RIS = 290), which suggested that the benefits observed in the current information set were conclusive. Sensitivity analysis showed that the heterogeneity of both “TZP 10 mg vs. TZP 5 mg” and “TZP 15 mg vs. TZP 5 mg” was derived from Rosenstock2021. There was no significant change in the effect size after removing this study, and the differences were still statistically significant, suggesting that the results were robust. The GRADE evaluation revealed that the quality of evidence for all indicators was high, with the exception of weight for “TZP 15 mg vs. TZP 5 mg”, which had a moderate quality of evidence. As shown in Compared with TZP 5 mg, meta-analysis showed that TZP 10 mg significantly reduced weight by 2.47 kg  and GI AEs by 17% , while serious AEs , MACE-4 , cardiovascular death , hypertension , cancer , hypoglycaemia (PG < 70 mg/dl)  and hypoglycaemia (PG < 54 mg/dl)  were comparable. TSA showed these outcomes did not reach the RIS or TSA threshold, and the results need more research and demonstration. In comparison to TZP 5 mg, meta-analysis indicated that TZP 15 mg increased total AEs by 10% , while serious AEs , GI AEs , MACE-4 , cardiovascular death , hypertension , cancer , hypoglycaemia (PG < 70 mg/dl)  and hypoglycaemia (PG < 54 mg/dl)  were comparable. TSA showed that cumulative Z values of total AEs exceeded the required information size at the fourth study , suggesting that the risk observed with the current information size was conclusive. Meta-analysis demonstrated that total AEs , serious AEs , GI AEs , MACE-4 , cardiovascular death , hypertension , cancer , hypoglycaemia (PG < 70 mg/dl)  and hypoglycaemia (PG < 54 mg/dl)  were comparable for TZP 15 mg compared with TZP 10 mg. TSA showed that the results observed by the current information volume are not conclusive. The results of the meta-analysis, TSA, and quality of evidence are shown in Compared with TZP 5 mg, meta-analysis revealed that TZP 10 mg increased total AEs by 6%   .Currently, metformin is still the first-line drug used for the initial treatment of T2DM , and it TZP is a novel GLP-1R and GIPR agonist. Previous meta-analyses have shown that TZP is significantly more effective than GLP-1RA and insulin in lowering glycemia and reducing weight, and does not increase total AEs or cardiovascular risk , 53, proMeta-analysis indicated that HbA1c, FSG and weight were significantly lower in the high-dose group of TZP than those in the low-dose group. TSA demonstrated that there was conclusive evidence for all efficacy endpoints except for the FSG of “TZP15 mg vs. TZP10 mg.” This implies that within the 5–15 mg dose, the effect of TZP in lowering glycemia and reducing weight is dose-dependent, and TZP 15 mg can achieve the best efficacy benefit in patients with T2DM. It is important to note that because of the presence of basal medication in 5 of the included studies –29, the Being overweight or obese is a major risk factor for T2DM  and insuIn the safety analysis, the tables of AEs were relatively similar across the included studies. They accurately measured AEs such as hypoglycemia, hyperglycemia, hypertension, pancreatitis, and gallstones, but the incidence of these AEs was low and did not differ statistically. In terms of safety endpoints, meta-analysis showed no statistical difference in serious AEs, MACE-4, cardiovascular death, hypertension, cancer and hypoglycaemia between the three dose groups of TZP. This implies that serious AEs, cardiovascular AEs, and hypoglycemic events may not be related to the dose of TZP. Meta-analysis demonstrated no significant differences in total AEs, serious AEs, GI AEs, or hypoglycaemia between TZP 15 mg and TZP 10 mg, suggesting that the safety profiles of high-dose TZP and medium-dose TZP are comparable. Compared with TZP 5 mg, the total AEs is significantly increased in TZP 10 mg and TZP 15 mg, which means that low-dose TZP has better overall safety. It is important to emphasize that most of the included studies had basal medication and therefore the safety results are primarily based on the context of the combination and not on the independent effect of TZP. Interestingly, the results of this study showed a significant increase in GI AEs for TZP 10 mg compared to TZP 5 mg, while TZP 15 mg had comparable GI AEs. Sensitivity analysis displayed that the heterogeneity of “TZP 15 mg vs. TZP 5 mg” was derived from the study of Frias2018 . The hetAlthough this study strictly followed the PRISMA guidelines for systematic reviews and meta-analysis, it continues to have some limitations. The first aspect is that neither the Del2021  nor the Given the limitations of the current study, future studies will be further improved. Firstly, increase the total sample size and use a rigorous randomized controlled double-blind trial. This not only improves the precision of the study, but also reduces potential methodological heterogeneity and increases the credibility of the results. Secondly, open research centers in countries with a predominantly people of African descent. This could explore the effects of TZP in populations of African descent and combine previous studies to synthesize the benefits and risks when using TZP in different ethnic groups. Thirdly, broaden the scope of efficacy indicators. 2-h postprandial glucose, lipids, blood pressure, body mass index (BMI), and cognitive function could be included as outcome indicators, which would allow a more comprehensive exploration of the combined benefits of TZP. Fourthly, control for relevant variables and perform a stratified study. A more comprehensive stratified analysis could be performed based on baseline information such as age, disease course, BMI, HbA1c level, and medication background, and thus learn about the efficacy and safety of TZP in different contexts. Fifthly, research in depth the properties of GIPR and the hypoglycemic potential of its activators. It has been shown that GIP stimulates insulin release to exert hypoglycemic effects under hyperglycemic conditions , while sThere are also a number of TZP-related clinical studies currently in the planning or conducting stage. NCT04657003 is a multi-center clinical study. The study will be conducted in various countries and regions, including the United States, Brazil, Japan, India, Russia, Puerto Rico, and Taiwan, and it is planned to include a sample size of approximately 900 patients to explore the efficacy and safety of TZP for the treatment of patients with obese or overweight with T2DM when measured by weight, BMI, waist circumference, HbA1c, FSG, lipids, blood pressure, and fasting insulin. NCT04660643 is a multi-center clinical study involving more than 70 countries and regions with a similar study topic to NCT04657003 and is expected to be completed in May 2023. Interestingly, NCT05024032 will evaluate the efficacy and safety of TZP in non-T2DM patients who are obese or overweight with weight-related comorbidities, and this randomized, controlled, double-blind trial will be conducted in China and is expected to include a sample size of 210. NCT04184622 will study the role of TZP in weight maintenance or weight loss following a program of intensive lifestyle changes, and they plan to collect a sample size of 2539 in 170 countries and regions. The two studies focused on the weight-reducing effects of TZP, rather than the effect of hypoglycemia, and may help broaden the indications of TZP. We are looking forward to the publication of the data from these studies and expect that the findings will be of benefit to patients with T2DM and the populations with obesity.TZP 15 mg > TZP 10 mg > TZP 5 mg in terms of lowering glycemia and reducing weight. TZP 5 mg > TZP 10 mg = TZP 15 mg in terms of safety. On this basis, we recommend TZP 5 mg as the first-choice dose for patients with T2DM to minimize the risk of AEs while reducing glycemia and weight. If patients cannot effectively control their glycemia after taking TZP 5 mg, it is recommended to take TZP 15 mg directly to achieve the best effect of glycemic reduction. Nevertheless, the independent efficacy and safety at different doses of TZP remains to be tested, as most of the included studies have a background of basal medication.The original contributions presented in the study are included in the article/supplementary material, further inquiries can be directed to the corresponding author.YY conceived and designed the study. GH and XY participated in data processing and statistical analysis. YY, GH, and SY drafted the manuscript. MZ, WJ, XY, and SY contributed to data analysis and interpretation. YY, MZ, and WJ supervised the review of the study. All authors seriously revised and approved the final manuscript.This work was supported by National Natural Science Foundation of China (81973753) and Natural Science Foundation of Hunan Province (2018JJ3405).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.


This study is intended to assess the prevalence of depression and anxiety in individuals who had recovered from COVID-19 and been discharged from hospital (RD hereafter) in Wuhan, China, and to explore the factors associated with these mental disorders.Patient Health Questionnaire, the Short Version of COVID-19 Stigma Scale, the Peace of Mind Scale, the Resilience Style Questionnaire, and the Perceived Social Support Questionnaire were used to collect relevant information of the participants. Descriptive analyses, Pearson correlation analysis, and logistic regression analysis were used to describe and analyze the data and to examine the factors associated with the mental health status of this population.Participants of this study were the RD who were infected at the beginning of the outbreak from 13 communities in Jianghan District of Wuhan City, Hubei Province, China by convenience sampling in mid-2021. The Generalized Anxiety Disorder Questionnaire, the In total, we recruited 1601 participants from 3059 COVID-19 patients, and 1541 participants completed the questionnaire survey, with a response rate of 96.25%. Finally, 1297 participants met the inclusion and exclusion criteria in this study, of whom 28.8% and 37.9% reported mild to severe levels of anxiety and depression symptoms. Perceived better mental health status during hospitalization, higher frequency of alcohol use per week, peace of mind, higher education level, and resilience were negatively associated with anxiety, while stigma and history of psychological or emotional counseling before infection was positively associated with anxiety. More severe clinical classification of COVID-19 and stigma  were both positively associated with depression, while perceived better mental health status during hospitalization , higher frequency of alcohol use per week , peace of mind , and social support  were negatively associated with depression.Tailored interventions on reducing stigma, enhancing mindfulness and social support should be taken into account to alleviate anxiety and depression among RD. As a major public health threat in the 21st century, the coronavirus disease 2019 (COVID-19) pandemic has caused tremendous mental health impact worldwide, including among those ever infected with the virus at the beginning of the outbreak .The uncertainties affected by the mass lockdowns and economic recession, and the lingering ‘long covid’ illness , along with the stigma they confronted from the surroundings, may cause mental disorders and even suicide among the RD population . The NatReports suggested that psychological problems among RD have multiple risk and protective factors. Discrimination and stigma due to the experience of infection were common among RD , 20. A sSince psychological distress is common among RD, it is of importance to identify the psychological symptoms and the associated factors among this population . HoweverThe cross-sectional study was carried out among former COVID-19 patients in Jianghan District  from June 10 to July 25, 2021. According to the electronic medical records of the Health Bureau of Jianghan District and inclusion criteria, a total of 3059 COVID-19 patients were eligible for the study and they were infected with the original SARS-Cov-2 strain and were diagnosed between December 10, 2019 and April 20, 2020. Among them, 1601 COVID-19 survivors were invited for a questionnaire survey on their mental health status when they were receiving clinical re-examination, and 1541 completed the survey and were included in the study, with a response rate of 96.25%. All investigators and support staff in this study were trained based on the same protocol and were required to have an educational background in medicine or public health. Those who had a history of COVID-19 infection, and ever have been hospitalized and discharged were invited to complete an online structured questionnaire from June to July, 2021. Digital informed consent was obtained from all individuals to ensure their voluntary participation. Self-administered electronic questionnaires and digital consent were sent to patients through Redcap, an online survey platform that is used for delivering online questionnaire. After reading the informed consent form and clicking the “I agree to participate in this study” button on the first page, participants could continue to fill in the electronic questionnaire. Ethics approval for the questionnaire survey was obtained from the Ethics Review Committee of the Institute of Pathogen Biology, Chinese Academy of Medical Sciences, Beijing, China (IPB-2020-22). The participants had to meet the following criteria: 1) over 18 years old; 2) had a history of hospitalization due to COVID-19 infection; 3) proficiency in Chinese; 4) able to independently cooperate with researchers to complete various scale assessments; 5) had a mobile communication equipment such as a mobile phone, and a WeChat account; 6) able to access the Internet with mobile equipment at any time; 7) had not received interventions for mental disorders within 1 month prior to enrollment or currently in the study. Those who met any of the below criteria were excluded: 1) had serious cognitive impairment; 2) had serious heart, brain, lung, kidney, liver, and other medical diseases or tumors; 3) difficult to cooperate with the questionnaire study. In total, 1297 participants were included in the final analysis based on the criterion above.Demographic characteristics, including age (“≤60” or >”60”), gender (“Man” or “Woman”), marital status(“Married” or “Unmarried/divorced /widowed”), income for 2020 (“<60000 yuan” or “≥60000 yuan”), dwelling state , education level (“Senior high school or below” or “Above senior high school”); and items on COVID-19 infection, such as family members’ infection, friends’ infection, length of hospital stay (“≤20 days” or “>20 days”), experience at ICU, clinical classification of COVID-19 , perceived mental health status during hospitalization , etc. were asked.The 7-item Generalized Anxiety Disorder Questionnaire (GAD-7) consists of 7 items that are rated on a 4-point Likert scaled from 0–3. It was developed for measuring the severity of generalized anxiety symptoms during the past two weeks . The scoPatient Health Questionnaire (PHQ-9) is a 9-item questionnaire that is used for screening and monitoring depression of varying degrees of severity during the past two weeks [The 9-item wo weeks . The itewo weeks , 31. In The Short Version of COVID-19 Stigma Scale is a 12-item scale that is employed for evaluating the perceived stigma of patients of COVID-19 during the past two weeks . Each itThe Peace of Mind Scale (PoM) comprises a total of 7 items rated on a 5-point scale ranging from 1  to 5  and is used for measuring the peace of mind during the past two weeks . Higher The Resilience Style Questionnaire (RSQ) consists of 16 items that are rated on a 5-point Likert scaled from 1 to 5. It is used to measure the level of individual’s resilience during the past two weeks . Higher The level of perceived social support of the participants was measured by two items including emotional support and material support during the past two weeks . The iteDescriptive analyses were performed to describe the participants’ demographic and clinical characteristics, the prevalence of psychological problems, and potential influencing factors. Pearson correlation analysis was employed to estimate the association between psychological variables. Univariate logistic regression analysis began with the full set of demographics to evaluate their associations with anxiety and depression. As suggested by previous research, the type I error rate could be relaxed in univariate logistic regression to avoid omitting possible influencing factors. Hence, statistically significant variables at the level of p≤0.10 in the univariate analysis as well as stigma, peace of mind, resilience, and social support were further entered into the multivariate logistic regression analysis . In the Among the 1297 participants, 47.3% aged above 60 years old; 56.6% were men; 85.2% were married; 62.1% had an annual family income of less than RMB 60000 in 2020; 12.2% lived alone; 29.0% were at an educational level of undergraduate or above; 46.5% had other family members infected with COVID-19; 29.1% had friends or relatives infected with COVID-19; 47.1% were hospitalized for over 20 days; 3.6% used to be treated at intensive care unit (ICU); 13.7% were in critical condition after infection; 56.7% perceived good psychological status during infection; 9.9% had a history of psychological or emotional counseling before infection; 12.4% were current smoker; 9.0% drank more than 2 times a week. The results are displayed in In this study, 28.8% and 37.9% of the participants reported mild to severe levels of anxiety and depression symptoms according to the cut-off values of GAD-7 and PHQ-9, as shown in The associations of anxiety, depression, stigma, resilience, peace of mind, and social support among the RD were all statistically significant at a 0.01 p-value, where anxiety and depression were all positively associated with stigma, and negatively associated with resilience, peace of mind, and social support. The results are illustrated in The factors associated with anxiety and depression of the participants are presented in Tables In multivariate analysis, we conducted the diagnostic evaluation of the model, the samples were all independent and the variables showed no multicollinearity. The multivariate analysis revealed that perceived better mental health status during hospitalization , higher frequency of alcohol use per week , and peace of mind  were negatively associated with anxiety, and a non-significant trend was observed in negative associations between anxiety and higher education level , and resilience . While stigma  was positively associated with anxiety, and non-significant trends were observed in positive associations between anxiety and history of psychological or emotional counseling before infection . Moreover, more severe clinical classification of COVID-19  and stigma  were both positively associated with depression, while perceived better mental health status during hospitalization , higher frequency of alcohol use per week , peace of mind , and social support  were negatively associated with depression.Mental disorders among RD are associated with their prognosis and may lead to more severe psychological sequelae without effective control . Hence, Based on the current study sample, we found that COVID-19 related stigma was positively associated with the occurrence of anxiety and depression among RD. Stigma has become a public health challenge during the COVID-19 pandemic and might cause severe mental illness in RD. Infected individuals who perceived being stigmatized could report guilt, self-isolation, self-depreciation, the experience of social exclusion, stereotyping, and insulation, hence they often suffer from psychiatric comorbidities, such as PTSD, anxiety, depression, and even suicidal ideation , 46. StiIn addition, the results of this study suggested that RD who had more severe clinical classification during hospitalization were more likely to develop depression. This finding is similar to the result of a multi-national cohort study in RD across six countries . LiteratBy contrast, we found that perceived better mental health status during hospitalization and social support were protective factors against mental disorders among RD. Consistent with previous researches, RD who could perceive good mental health status usually viewed negative life events more optimistically, and could adapt themselves better to new lives after discharge, thus reducing the possibility of having psychological problems , 50. HigAdditionally, findings of this study indicated that peace of mind and resilience were negatively related to mental disorders among RD. Previous research has suggested that peace of mind is associated with anxiety and depression . IndividAlthough previous systematic reviews indicated substantial harm of alcohol use on health, the current study found alcohol use could protect RD from both anxiety and depression . A crossMoreover, the findings of this study indicated that RD with a history of psychological or emotional counseling before infection might be at higher risk of anxiety. Evidence suggested that individuals with a history of psychological counseling might have experienced psychological distress before, and the distress was more likely to reoccur due to COVID-19 infection among RD , 79. ThiOverall, findings of this study suggested that the government, health authorities, as well as social media could consider alleviating mental disorders among RD from several dimensions, such as mobilizing society to stop stigmatizing, providing more support for RD, and implementing psychological interventions in RD.This study has several limitations. First, a drawback of the cross-sectional study is that we can’t make any statements regarding causality. As a consequence, findings on risk factors of adverse psychological outcomes should be accepted with caution. Second, our study was conducted more than 18 months after discharge of COVID-19 patients, which may lead to recall bias. Third, convenience sampling may decrease the representativeness of the population, and cause biased calculation of confidence interval in the logistic regression analysis. Fourth, the results may be biased due to various access to technology, connectivity, and skills to use mobile communication equipment. Fifth, we did not consider the heterogeneity between 13 communities of the jianghan district, which may cause bias. Sixth, in the categorization of some demographic variables, such as income and length of hospital stay, we set the cutoff point based on the data characteristics, which may be a little subjective. Seventh, in the evaluation of alcohol use, we only consider the drinking frequency without alcohol volumes, which may not accurate enough to assess alcohol use. Eighth, we recruited participants who had not received interventions for mental disorders within 1 month prior to enrollment without considering the bias caused by having received any type of psychological intervention two or more months prior to the survey. In addition, our study didn’t collect mental health information about patients who died of COVID-19, which may underestimate the rate of mental health problems in discharged patients infected with COVID-19.The prevalence of anxiety and depression in people who had recovered from COVID-19 and had been discharged from hospital in Wuhan, China were 28.8% and 37.9% in this study. We found that stigma caused by COVID-19 infection was positively associated with mental disorders, while social support, peace of mind, psychological resilience, and alcohol use were negatively associated with mental disorders like anxiety and depression in RD. More attention should be paid to RD with a history of psychological or emotional counseling before infection. Tailored interventions regarding reducing stigma, enhancing mindfulness level and social support should be taken into account to alleviate anxiety and depression among RD. Further research could focus on prospective studies or long-term follow-up to examine the causal relations between the factors identified in this study.S1 Data(XLSX)Click here for additional data file.