Mucosa-associated lymphoid tissue (MALT) lymphoma is the most common subtype of marginal zone lymphomas. These B-cell neoplasms may arise from many organs and usually have an indolent behavior. Recurrent chromosomal translocations and cytogenetic alterations are well characterized, some of them being associated to specific sites. Through next-generation sequencing technologies, the mutational landscape of MALT lymphomas has been explored and available data to date show that there are considerable variations in the incidence and spectrum of mutations among MALT lymphoma of different sites. Interestingly, most of these mutations affect several common pathways and some of them are potentially targetable. Gene expression profile and epigenetic studies have also added new information, potentially useful for diagnosis and treatment. This article provides a comprehensive review of the genetic landscape in MALT lymphomas.TNFAIP3, CREBBP, KMT2C, TET2, SPEN, KMT2D, LRP1B, PRDM1, EP300, TNFRSF14, NOTCH1/NOTCH2, and B2M, but many other genes may be involved. Similar to chromosomal translocations, certain mutations are enriched in specific lymphoma types. In the same line, variation in immunoglobulin gene usage is recognized among MALT lymphoma of different anatomic locations. In the last decade, several studies have analyzed the role of microRNA, transcriptomics and epigenetic alterations, further improving our knowledge about the pathogenic mechanisms in MALT lymphoma development. All these advances open the possibility of targeted directed treatment and push forward the concept of precision medicine in MALT lymphomas. Mucosa-associated lymphoid tissue (MALT) lymphomas are a diverse group of lymphoid neoplasms with B-cell origin, occurring in adult patients and usually having an indolent clinical behavior. These lymphomas may arise in different anatomic locations, sharing many clinicopathological characteristics, but also having substantial variances in the aetiology and genetic alterations. Chromosomal translocations are recurrent in MALT lymphomas with different prevalence among different sites, being the 4 most common: t, t, t, and t. Several chromosomal numerical abnormalities have also been described, but probably represent secondary genetic events. The mutational landscape of MALT lymphomas is wide, and the most frequent mutations are:  Helicobacter pylori (H. pylori) recently described in several studies [Mucosa-associated lymphoid tissue (MALT) lymphoma was first described in 1983 by Isaacson and Wright in the stomach but may arise from other mucosal tissues , MALT ly studies .MALT lymphomas mainly occur in adults, with a median age about 60 years. Men and women are affected equally, although there is site specific female predominance in the parotid gland and breast ,8. The mH. pylori infection, satisfying Koch’s postulates for an etiologic agent [Helicobacter heilmannii in the stomach [Chlamydia psittaci (C. psittaci) in the ocular adnexa [Borrelia burgdorferi in the skin [Campylobacter jejuni in immunoproliferative small intestine disease [Achromobacter xylosoxidans in the lung [Gastric MALT lymphoma is linked with chronic ic agent . Albeit  stomach , Chlamydr adnexa ,13, Borrthe skin ,15, Camp disease  and, Achthe lung . Autoimmthe lung . There athe lung .MALT lymphomas are considered indolent neoplasms with 5-year relative survival rate of 93.8%, higher than that observed in SMZL (85.3%) and NMZL (82.8%) . Among MHigh throughput genome-wide methodologies, the complete sequencing of the human genome and recent developments in next generation sequencing (NGS) have allowed for unprecedented insights into the genomic alterations that underlie oncogenesis, tumor biology, and survival. Since more than 95% of patients with hematologic malignancies have adequate tissue for genomic profiling, this represents an excellent opportunity to improve diagnosis and classification, to look for prognostic markers and also for detection of pharmacologically tractable targets . This reA functional B-cell receptor (BCR) is essential for the biology of B-cells. MALT lymphoma cells almost always express surface immunoglobulin (Ig) M and its BCR signalling is functional. This is supported by their proliferative responses to mitogens  and the MALT lymphomas have highly altered variable heavy chain immunoglobulin (IGHV) and variable light chain immunoglobulin (IGLV) genes, which are consistent with germinal center (GC) or post-GC origin ,23,24. AThe variation in Ig gene usage among MALT lymphoma of different anatomic locations is presumably the result of adaptive response and clonal selection induced by their varied aetiologies, resulting in different antigen exposures . The Ig In MALT lymphomas of the salivary gland (SGMZL), there is a clear skewed usage of IGHV1-69/J4 (55%) or IGHV3-7/J3 (15%) rearrangements, and this together with other less frequent (IGHV4-59/J2(J5) and IGHV3-30/JH4) rearrangements indicates that most salivary gland MALT lymphomas express BCR that potentially bears RF activities ,33.C. psittaci infection. C. psittaci-negative cases have a much greater prevalence of IGHV4-34 usage than those C. psittaci-positive [TNFAIP3 inactivation by deletion or mutation was significantly higher in MALT lymphomas with IGHV4-34 rearrangement (54%), particularly in those of OAMZL (70%), than in those using other IGHV genes (20%). The concurrence of IGHV4-34 rearrangement and TNFAIP3 inactivation points toward a cooperative relationship of between these two events in OAMZL lymphomagenesis [IGHV4-34 (18%) is the most often usage in ocular adnexal MALT lymphoma (OAMZL), followed by IGHV3-23 (12–17%), IGHV3-30 (10–14%), and IGHV3–7 (9%) ,37,38,39positive , raisingH. pylori antibiotic treatment and without the t of IGHV3-30 or IGHV3-23. MALT lymphomas of the lung and skin have also found to have biased usage of IGHV3 or IGHV4 and IGHVH1-69 or IGHVH4-59, respectively [MALT lymphomas from other sites have also biased usage of IG genes, albeit those coding for autoantibodies are still overrepresented. Gastric MALT lymphomas (GMZL) have a biased use of IGHV4-34, IGHV3-7 and IGHV1-69 genes ,44,45,46ectively . Moreoveectively ,48. HoweThe identification of cytogenetic abnormalities constitutes an important tool for establishing the diagnosis, monitoring the clinical course, and assessing the prognosis of patients with B-cell lymphomas. Cytogenetic analysis in MALT lymphomas was initially hampered by the facts that biopsies are usually small (most taken by endoscopic procedures) and then rarely subjected to conventional cytogenetic analysis and also, because its proliferation in vitro is often poor. Then, during several years, data were limited in comparison with other types of indolent B-cell lymphomas. However, the application of FISH (fluorescence in-situ hybridization), SKY  and high-resolution technologies such as array based comparative genomic hybridization (array-CGH) have improved our knowledge of chromosomal abnormalities in MALT lymphomas. In the last two decades, a variety of chromosomal structural and numerical alterations have been described in MALT lymphomas . ChromosThe most common recurrent translocation is t which juxtaposes the N-terminal region API2 gene, containing 3 BIR domains with inhibitor caspase activity, and the C-terminal region of MALT1 gene, containing an intact caspase p20-like domain ,57,58,59t is most frequent in MALT lymphomas of the lung (40%) and stomach (20–25%). This translocation is also found in the intestinal  and secondary forms (57%), but rare in colorectal cases), the ocular adnexa (10%), and it is uncommon or not present in MALT lymphomas of the thyroid, salivary gland, and skin ,70,71,72H. pylori antibiotic treatment, but only in 3% of those that respond to H. pylori eradication [In the MALT lymphomas of the stomach, t is found in 47% and 68% of cases with stage IE and stage IIE or above, respectively, which do not respond to dication ,85,86,87dication . Additiodication . Howeverdication  or in codication  or bendadication . For thedication ,94. Patients with t had more frequent monoclonal gammopathy, in particular of IgM subtype (31% v 8%), some of which developed class switch ,95. The MALT1 gene under the transcriptional control of the IgG enhancer, then MALT1 expression is deregulated fostering NF-kB activation [IGH/MALT1 show not only over-expression of MALT1, but also BCL10 accumulation in cytoplasm. This finding suggests that MALT1 may immobilize BCL10 in cytoplasm through their interaction [The t is the second most common among all balanced translocations in MALT lymphoma and brings the tivation ,102,103.tivation ,106,107.eraction . This reeraction ,110. Otheraction ,112,113.eraction ,115.BCL10 gene under the regulatory control of the IGH gene, resulting in deregulated expression [BCL10-IGH. Over-expression of BCL10 and formation of CBM signalosome via oligomers uniting the CARD domain results in NF-κB activation [The t juxtaposes the pression . In normpression . Howevertivation .H. pylori eradication [t is particularly associated with MALT lymphomas, albeit it is infrequent (1–2% of MALT lymphomas). This translocation may be seen in MALT lymphoma of the lung (9%) and stomach (4%), but it is rare in other sites: ocular adnexa, salivary gland, thyroid and skin ,67. The dication ,85.IGH and FOXP1, is found in approximately 10% of MALT lymphomas, mainly from cases arising in the thyroid, ocular adnexa and skin. MALT lymphomas from the stomach and the lung, NMZL and SMZL are typically negative. Most MALT patients carrying t also harbor additional genetic abnormalities, such as trisomy 3 [t, with rearrangement of risomy 3 .FOXP1-IGH [CNN3-IGH, t/ODZ2-IGH, t/JMJD2C-IGH [GPR34-IGH [IGH gene locus, (or with the kappa light chain gene) and cause their over-expression. For instance, FOXP1 over-expression though inhibition of apoptosis and plasma cell differentiation may contribute to the pathogenesis of MALT lymphomas [FOXP1-IGH is found in 7–56% of thyroid MALT lymphoma cases, [FOXP1-IGH may be useful for the differential diagnosis between primary MALT lymphoma of the thyroid and other thyroid disorders [GPR34 mutation [Several novel chromosome translocations in occasional cases of MALT lymphoma have been described: t/OXP1-IGH ,119,120,JD2C-IGH , tPR34-IGH ,123 and PR34-IGH ,124. Theymphomas ,126. Thea cases, ,127,128,mutation ,123.Beyond translocations, a spectrum of chromosomal numerical abnormalities has been described in MALT lymphomas. In the first cytogenetic study in MALT lymphomas from the Isaacson’s group, numerical abnormalities of chromosomes 3 and 7 were found in a series of 23 MALT lymphomas . CurrentThe most common numerical alterations found in MALT lymphomas are trisomy of chromosome 3 or 18, although the frequencies at which these trisomies occur vary markedly with the primary site of disease ,131. In BCL6 [FOXP1 [CCR4 [BIRC3-MALT1 rearrangement [H. pylori eradication treatment in one study [The genetic mechanisms by which trisomy 3 contributes to lymphomagenesis are not fully clarified. However, the biological effects of chromosomal trisomies are likely to be explained by an enhanced gene dosage effect resulting from larger copy numbers of genes crucial to lymphoma development. Several candidate genes, such as the protooncogene BCL6 , the tra6 [FOXP1  or the cP1 [CCR4 , all fouangement . Of notene study .Trisomy 18 has an approximate frequency around 20% ,147. ItsOther trisomies, such as trisomies 7, 12 and others, have been observed non-randomly but less frequently than trisomy 3 or 18. It is worth mentioning that different from other lymphoma types, copy-neutral LOH did not appear to be a common event in MZL .C-MYC are reported to have rapid disease progression [TP53 have also been described in a considerable proportion of gastric MALT lymphomas [Improvements in the cytogenetic tools have allowed the identification of lesions with relevant role in the lymphomagenesis of MALT lymphomas. Structural aberrations of chromosome 1 frequently involved the chromosomal regions 1p22, 1p34 and 1q21 ,148 and gression . C-MYC agression . Alteratymphomas ,150 and ymphomas .TNFAIP3 (A20) was detected in MALT lymphomas of the ocular adnexa (19%), thyroid (11%), salivary gland (8%), and liver (0.5%), but not in the lung, stomach, and skin [TNFAIP3 inactivation is associated with reduced lymphoma-free survival [TNFAIP3 inactivation alone is not sufficient for malignant transformation but nonetheless may represent a promising future therapeutic target. In addition, deletion of CD274 (PD-L1) are frequently found in MALT lymphomas of the thyroid, together with mutation in up to 68% of cases (see later) [Other genomic imbalances have also been found using array-CGH ,151,152.and skin ,155,156.survival ,157. TNFsurvival . Howevere later) .Although mutational profiling of hematologic neoplasms currently predominates in the area of myeloid neoplasms, there is a growing wealth of literature describing common and highly characteristic genetic alterations in lymphomas. Certain mutations are enriched in specific lymphoma types, and new discoveries continue to emerge at a rapid pace. TNFAIP3 , CREBBP (22%), KMT2C (19%), TET2 and SPEN , KMT2D, LRP1B, and PRDM1 , EP300 , TNFRSF14 , NOTCH1/NOTCH2  and B2M  [GPR34 (1–19%) and CCR6 (1–8%) [KMT2C, KMT2D, CREBBP and EP300 tended to coexist [The mutational landscape of MALT lymphomas is wide and, in the last decade, has been characterized using high-throughput technologies such as whole exome sequencing, whole genome sequencing and customized NGS panels. The data available to date show frequent lesions affecting chromatin remodeling and transcription regulation, BCR and NF-κB signalling, NOTCH pathways and immune surveillance. Not surprisingly, MALT lymphomas show considerable variations in the incidence and spectrum of genetic mutations among different sites, similar to what happens with translocations. Overall, the most frequent mutations are: eletion) . Other m6 (1–8%) . Similar coexist   inactivation by mutation and/or deletion is frequent in MALT lymphomas, in particular in those arising in the ocular adnexa (29–54%) [TNFAIP3 mutations are deleterious changes (frameshift indels and nonsense mutations), resulting in a truncated protein. Interestingly, TNFAIP3 mutations rarely co-existed with mutations with other genes, supporting their important pathogenetic role in these lymphomas [TNFAIP3 truncation mutants show a substantial impairment in repression of NF-kB activation. TNFAIP3 is a key regulator of inflammation signaling pathways as well as a negative regulator of the NF-κB pathway, which inhibits NF-κB activity triggered by signaling from a variety of surface receptors [TNFAIP3 inactivation can increase the activation of the canonical NF-κB pathway triggered by signalling from several receptors including BCR, TLR and TNFR1 due to loss of negative regulation on several signalling molecules  downstream of their receptors [TRAF3 inactivation promotes the activation of the non-canonical NF-κB pathway due to impaired control on NIK degradation [TRAF3 and TNFAIP3 were mutually exclusive [TNFAIP3 and MYD88 mutations, MALT lymphoma shows rare or no mutations in other members of NF-kB pathway such as CD79A, CD79B, CARD11, BIRC3, and TNFRSF11A, which are frequently seen in other B-cell lymphomas with constitutive NF-kB activation. As previously mentioned, (29–54%) ,167,168,(29–54%) , salivar(29–54%)  and thyr(29–54%) , among oymphomas . TNFAIP3eceptors . Thus, Teceptors ,173,174.radation . Thus, ixclusive . Beyond NOTCH1 and NOTCH2, but mainly in MALT lymphomas of the ocular adnexa (8–10%) [NOTCH1/2 product lacking TAD or PEST domains, which governs protein stability and degradation [NOTCH1 mutations were enriched in patients who failed to H. pylori eradication [NOTCH1/2, genetic lesions in additional NOTCH pathway modulators or members, including SPEN (17%) and DELTEX1, have been identified [NOTCH stability and activity [NOTCH signalling regulates multiple aspects of lymphoid development and function. Although NOTCH inhibits the earliest stage of B lymphopoiesis, it is nevertheless necessary for the development and maintenance of marginal zone lymphocytes ,178,179. (8–10%)  and dura (8–10%) . These mradation . NOTCH1/radation ,185,186,dication . In addientified . These mactivity .TBL1XR1, TET2, MLL2/KMT2D and CREBBP [TBL1XR1 mutations are found in MALT lymphoma of the salivary gland (24%) [H. pylori-resistant gastric MALT lymphoma (16%) [TBL1XR1 mutations were only seen in association with NOTCH2 mutations , which might indicate a co-operative role for these mutations in lymphomagenesis [TBL1XR1 mutations are missense changes affecting regions or residues critical for interaction with NCoR, and may increase TBL1XR1 binding to NCoR and facilitate its degradation, consequently promoting NF-κB and JUN target gene expression [TET2 mutations are frequently found in MALT lymphoma of the thyroid (62%) [MLL2/KMT2D mutations have been described in several B-cell lymphomas, and the frequency in MALT lymphomas is variable among studies (5–25%) [CREBBP mutations are also frequent observed (4–22%) [Several chromatin remodeling and transcriptional regulators are recurrently mutated in MALT lymphoma of different sites, including d CREBBP ,161,176.nd (24%) , ocular nd (24%) ,188, andagenesis . The majpression ,159. TETid (62%)  and will (5–25%) ,160,161. (4–22%) ,188,189.TNFRSF14 are frequently seen in those arising in the thyroid (46–52%) [TNFRSF14 mutations are deleterious changes such as nonsense, frameshift alterations, thus most likely inactivating or impairing the protein function. Thereby, TNFRSF14 inactivation may enable malignant B-cells to gain more T-cell help.Genetic abnormalities that regulate B and T cell interactions have also been described in MALT lymphomas. Mutations in (46–52%) ,159 but (46–52%) ,145,159.As previously discussed, mutations in MALT lymphoma can vary widely depending on the site of origin and, therefore, characteristic locations have been the subject of specific genetic studies. TNFAIP3 (30%), MYD88 (5–20%), and BCL10 (6–25%), whereas other genes were rarely or not mutated in the cases analyzed [KMT2D (22%), NOTCH1 (8%), NOTCH2 (8%), TNIP1 (5%), NFKBIA (4%), and in other genes at lower frequencies [TNFAIP3 mutations (36%), and also recurrent mutations in MYD88 (7%), TBL1XR1 (6%), TNFRSF14 (5%) and TET2 (4%), besides a few less frequently mutated genes [CREBBP (9–17%) [LRP1B (6%) [JAK3 (11%) that were located at sites known to alter the protein function and leading to activation of the JAK/STAT signaling pathway by causing a gain of function of JAK3 [JAK3 mutation had shorter progression free survival, but not reduced overall survival [JAK3 mutations in OAML opens the option for potential precision therapeutics by targeting the JAK/STAT pathway with JAK inhibitors. Other newly identified genes recurrently mutated (5–10%) included members of the collagen family (COL12A1 and COL1A2) and DOCK8 which is involved in RhoGTPase signaling [OAMZL have been the focus of mutation investigations by several groups. Initial studies found recurrent mutations in analyzed ,167,190 quencies . Using Wed genes . A furth (9–17%) ,168 and P1B (6%) . Interes of JAK3 . Patientsurvival . Identifignaling . TBL1XR1 = 24%, GPR34 = 19%) [GPR34 mutation is almost exclusive of this site and mutations in GPR34 and CCR6, both members of the GPCR family, are mutually exclusive [In SGMZL, G-protein coupled receptors have been found mutated at high frequency (4 = 19%) . Interesxclusive .TET2 (86%), and also mutations in CD274 (53%), TNFRSF14 (53%), and TNFAIP3 (30%) [TNFRSF14 and TET2 had a significantly higher variant allele frequency than TNFAIP3, suggesting that TNFAIP3 mutations may occur later than TET2 and TNFRSF14 changes. In patients carrying TET2 mutations, 46% of them had two mutations and also had a significantly higher number of somatic variants compared with those without. Most TET2 mutations are deleterious alterations, inactivating or impairing its dioxygenase activity, hence its role in DNA demethylation, consequently affecting a wide spectrum of gene expression [TET2 inactivation may deregulate the expression of transcriptional factors indispensable for B-cell function, and thus potentially cooperate with receptor signalling, including those by the enhanced T-helper cell signals, indirectly triggered by PD-L1/TNFRSF14 inactivation in malignant B-cells [In MALT lymphomas of the thyroid, Wu et al., have found frequent deleterious mutations of P3 (30%) . Both TNpression ,191. In  B-cells .H. pylori- negative gastric MALT lymphoma. In addition to reconfirming that the MALT1 translocation is the most frequent genetic alteration /BIRC3-MALT1) and IGH translocation was further seen in 40% of MALT1-negative cases, mutations in NF-kB signaling pathways were detected in 40% of cases . The NF-kB pathway mutations were mutually exclusive from MALT1, albeit not IGH translocation, in total occurring in 86% of cases [H. pylori- negative gastric MALT lymphomas harbor activating mutations that affect both canonical and non-canonical NF-kB pathways [Kiesewetter et al., have recently investigated the genetic characterization of of cases . Then, Hpathways .Other studies analyzing the mutational landscape of MALT lymphomas at several sites have been conducted and the list of involved genes keeps growing .MicroRNAs are small non-coding RNAs that modulate gene expression on a post-transcriptional level, playing critical roles in cellular proliferation, apoptosis and differentiation ,195. MicH. pylori-associated gastric MALT lymphoma, Craig et al., identified miR-203 as one of the most strongly downregulated miRNAs in comparison with normal lymphoid tissue, suggesting that might be a key player in the transformation from chronic gastritis to MALT lymphoma. In addition, since ABL1 is the target of miR-203, treatment with imatinib and dasatinib (BCR/ABL inhibitors) prevented tumor cell growth [C-MYC [In l growth . Zhang eh [C-MYC .H. pylori infection, may be a potential predictor of resistance to H. pylori eradication therapy, independently of API2/MALT1 fusion gene [H. pylori eradication [miR-142-5p and miR-155 have also been found overexpressed in gastric MALT lymphoma . miR-155ion gene . Inhibitdication ,202.Our group found through analyses of expression of 384 miRNAs that gastric MALT lymphomas are characterized by specific miRNA expression profile, different to that seen in chronic gastritis and reactive lymphoid tissue. In addition, 17 differentially miRNAs were expressed between patients carrying or not t. Overexpression of miR-142-3p and miR-155 and downregulation of miR-203 were detected in gastric MALT lymphomas in comparison with chronic gastritis. These finding suggest that expression levels of these miRNAs might be useful for the differential diagnosis between chronic gastritis and gastric MALT lymphoma [In conjunctival MALT lymphoma, some upregulated miRNAs, such as miR-150/155, as well as downregulated miRNAs, such as miR-184, miR-200a/b/c, and miR-205, have also been identified. Dysregulation of the miR-200 family might be involved in the pathogenesis and progression of the lymphoma . The miRAll of these miRNAs may be important in the pathomechanisms of MALT lymphoma. However, further studies are needed to clarify their role in the pathogenesis and also its utility in the clinical practice.Understanding the transcriptome, the complete set of transcripts in a cell and their quantity, is crucial for interpreting the functional elements of the genome and revealing the molecular constituents of cells and tissues, and also for understanding development and disease . IntegraMALT1 translocations, having overexpression of NF-KB pathway genes and enrichment for chemokine signaling pathways . Another subset displayed increased plasma cells and a prominent plasma cell gene signature [MALT1 translocations with high expression of MALT1 and RARA, samples with plasmocytic differentiation having high FKBP11 expression, and cases with high RGS13 expression tending to have trisomy 3 and reactive follicles [The gene expression profile of MALT lymphomas of the lung was analyzed by Chng et al., who described a prominent T-cell signature and a marginal zone/memory B-cell profile . Gene exignature . In addiollicles . CCR4 (3ollicles .FOXP1 (3p14.1) is overexpressed in a few MALT lymphomas due to its juxtaposition to the immunoglobulin heavy chain gene promoter after chromosomal translocation [In 33 MZL cases (18 MALT lymphomas), gains affecting chromosomes 3q and 18 seemed to influence B-cell receptor signaling pathways, cell cycle, Wnt signaling, and apoptosis, whereas genes associated with 3p and 18p gains formed transcripts involved in chemokine and cytokine signaling pathway, ubiquitin proteasome pathway, Ras signaling, and in tight junction regulation . FOXP1  are characterized by an enhanced expression of NF-kB target genes, mainly and ICOS . The expfference .BIRC3-MALT1 fusion was analysed by Cascione et al. [TNFSF12-TNFSF13 (TWE-PRIL), TNFRSF17 (BCMA), LTB (lymphotoxin β), and CXCR3 were among the most upregulated genes in this subset of patients.Recently, the transcriptome of MALT lymphomas of the lung harbouring e et al. , identifH. pylori-positive gastric MALT lymphoma cells, Zou et al., have found 15 pathways differentially enriched, including the Wnt, the mTOR, the NOD-like receptor and the Hippo signalling pathways. By deregulating these pathways, H. pylori might influence gastric lymphomagenesis through modulating the proliferation of cells, inducing autophagy and fostering inflammatory responses and epithelial mesenchymal transition. By proteomic analysis, they also identified 116 differentially expressed proteins, most of them previously associated with cancer that can be used as biomarkers for lymphoma diagnosis or as potential therapeutic targets [In  targets .H. pylori-infected gastritis and gastric cancer or MALT lymphoma [H. pylori infection but also affect the risk of MALT lymphoma [Zhang et al., conducted a study to identify transcriptomic biomarkers in lymphoma . In gastlymphoma . In addilymphoma .TET2, KMT2D, CREBBP, TBL1X1, KMT2C, MT2C, EP300, among others [Epigenetic alterations such as disturbances of DNA methylation and histone modification are common in B-cell lymphomas, contributing to lymphomagenesis . Alteratg others ,169,176 TET2 mutations were very common with frequencies up to 85.5% [EZH2, EED and SUZ12) targets, genes bearing histone H3 dimethylation at K4 (H3K4me2) and trimethylation at K27 (H3K27me3). TET2 mutations have been also described (in percentages below 10%) in the salivary gland, ocular adnexa, stomach, and lung (among others). Since TET2 mutations are associated with higher response rate to demethylating agents in myeloid leukemias, it is reasonable to investigate the role of demethylating agents in patients with TET2 mutated MALT lymphomas.In thyroid MALT lymphomas, as previously mentioned, to 85.5% ,159. MutKMT2D with frequencies varying from 14% to 22% [Helicobacter pylori-negative gastric MALT lymphoma, KMT2D (17%) and CREBBP (14%) are the two most epigenetic regulator gene mutations [CREBBP are found in 4–22% of MALT lymphomas [CREBBP gene encodes a lysine acetyltransferase (KAT) protein that activates gene expression through acetylation of histone H3 lysine 18 (H3K18Ac), histone H3 lysine 27 (H3K27Ac), and other residues. KAT domain mutation of CREBBP inhibit its catalytic activity and leads to a dominant-repressive effect by preventing the participation of redundant acetyltransferases in transactivation complexes, leading to loss of antagonism to BCL6-mediated gene repression and reduced expression of antigen presentation and interferon signaling genes in germinal-center lymphomas [CREBBP mutant cells [CREBBP-mutant MALT lymphomas, alone or in combination with PD1/PD-L1 blockade (to prevent interferon-induced adaptive immune suppression).In OAMZL, the most commonly reported epigenetic mutated gene is % to 22% ,161,168,utations . In factymphomas ,189. Theymphomas . By usinnt cells . This suTP15 and TP73 genes was detected more frequently in gastric MALT lymphomas than in gastric or nodal DLBCL [DAP-k (72.2%), GSTP1 (50%), MGMT (27.2%) and TP73 (9%) have been found in MALT lymphomas [DAPK and INK4aTP16 was frequently observed at its initial presentation, but not at tumor progression [While classical mutations in genes are region-limited, epimutations often occur early in cancer development and have a genome-wide impact. CpG island hypermethylation of al DLBCL . Hypermeymphomas . Inactivymphomas . In MALTymphomas . The samL [DAP-k .2%, GSTPFurthermore, aberrant methylation has also been implicated in tumor progression. Dugge et al., investigated the DNA methylation changes associated with progression of gastric MALT lymphoma through genome-wide DNA methylation profiling and observed that 7698 CpG loci associated with 2419 genes were significantly differentially methylated during gastric MALT lymphoma progression . Among tSince epigenetic changes are potentially reversible, epigenetic therapy, both alone or in combination with established treatment regimens, has a promising future, leading to potentially less toxic and more effective approaches that standard chemotherapy.MYD88 mutations in distinguishing LPL from MZL or the presence of NOTCH2 and TNFAIP3 mutations in favoring a diagnosis of MZL over LPL [Davis et al., have explored the clinical and practical diagnostic utility of a targeted NGS within mature lymphoid neoplasms (MLN) . They stover LPL .EZH2 mutation and who have received at least 2 prior systemic therapies and also for relapsed or refractory follicular lymphoma who have no suitable alternative treatment options [Pillonel et al., reported a 3-year experience with a 68-gene lymphoma NGS panel applied to 80 cases . The ana options . This prCurrently, NGS analysis is not generally applied to patients as first-line investigational approach, but rather applicable to further define diagnosis in highly selected otherwise equivocal instances and/or to identify mutations that will guide treatment decisions in cases in which standard treatment options are scarce or suboptimal .MALT lymphomas are a diverse group of lymphoid neoplasms that exhibit a wide range of genetic features depending on the site of origin. Despite the fact that MALT lymphoma of different anatomic locations shares many common clinicopathological characteristics, there are substantial variances in the aetiology, Ig gene usage, and acquired genetic alterations. Then, it is crucial to better characterize the somatic mutation profile of MALT lymphomas and unravel their molecular oncogenic complexity applying the latest NGS platforms.Excitingly, the majority of the genetic changes affects NF-KB signal pathway-related genes, resulting in constitutive NF-kB pathway activation. At the same time, since targeted gene sequencing panels have been recently applied to MALT lymphomas, we have learned that other genes are also frequently affected such as those involved in chromatin remodeling, BCR/TLR and NOTCH pathways. The growing knowledge of its genetic landscape, methylation, and transcriptome features raises the possibility of applying new agents to treat a subset of MALT lymphoma patients and also push forward the concept of precision medicine in this type of lymphoma.


Background: Glioblastoma (GBM) is the most common and aggressive primary intracranial tumor of the central nervous system, and the prognosis of GBM remains a challenge using the standard methods of treatment—TMZ, radiation, and surgical resection. Traditional Chinese medicine (TCM) is a helpful complementary and alternative medicine. However, there are relatively few studies on TCM for GBM.Purpose: We aimed to find the connection between TCM and anti-GBM.Study design: Network pharmacology and inflammatory microenvironment strategy were used to predict Siraitia grosvenorii (Luo Han Guo) target for treating glioblastoma.Methods: We mainly used network pharmacology and bioinformatics.Results: CCL5 was significantly highly expressed in GBM with poor prognostics. Uni-cox and randomForest were used to determine that CCL5 was especially a biomarker in GBM. CCL5 was also the target for SG and TMZ. The active ingredient of Luo Han Guo — squalene and CCL5 —showed high binding efficiency. CCL5, a chemotactic ligand, was enriched and positively correlated in eosinophils. CCL5 was also the target of Luo Han Guo, and its effective active integrate compound –— squalene — might act on CCL5.Conclusion: SG might be a new complementary therapy of the same medicine and food, working on the target CCL5 and playing an anti-GBM effect. CCL5 might affect the immune microenvironment of GBM. Glioblastoma (GBM) is the most common and aggressive primary intracranial tumor of the central nervous system . Most ofSalvia miltiorrhiza can inhibit the proliferation of U87 cells, induce apoptosis, and enhance the efficacy of TMZ  were used for the treatment of various cancers , such asy of TMZ . Lycium  the BBB . Magnolo the BBB . HoweverSiraitia grosvenori (SG) is a perennial herbaceous plant of the Cucurbitaceae family with huge resource reserves and native to southern China, also known as monk fruit and Luo Han Guo, which is a medicinal food homologous species granted by the China Food and Drug Administration with significant clinical effects  for GBM  text mining, we selected the most effective clinical drug in the treatment of GBM. Based on the TCMSP (http://bioinformatics.psb.ugent.be/webtools/Venn/) was drawn for visualizing the SG-TMZ interacting targets. Gliomas-related targets were predicted by OMIM . Differentially expressed GBMs (DE-GBMs) were computed by limma  network, scores >0.70 were considered to have high confidence . FunctioHazard ratios (HR) were applied using univariable Cox (uni-cox) regression analysis . We then detected the key SG-TMZ-GBMs by “survival” and “survminer” package . Random https://www.proteinatlas.org/ENSG00000271503-CCL5/pathology/glioma#).The microenvironment cell population-counter method was chosen to evaluate the association between CCL5 and immune cell populations . We usedPubMed text mining showed 2121 literature reports on the treatment of GBM with TMZ . ThroughWe obtained 12 chemical compositions in SG by TCMSP and TCMID, and gained the compound structure by PubChem, and predicted the 474 target genes of SG by the Swiss Target Prediction and PharmMapper. A total of 113 SG-TMZ targets were found by taking the intersection . We furthelicobacter pylori infection,” Vascular Endothelial Growth Factor A (VEGFA), Rac Family Small GTPase 1 (RAC1), Protein kinase C Beta (PRKCB), and AKT Serine/Threonine kinase 1 (AKT1) were enriched in Vascular Endothelial Growth Factor (VEGF) signaling pathway (We imported 42 SG-TMZ-GBMs into the STRING database to construct a protein-protein interaction (PPI) network, the primary connection in the network which might have pharmacological effects in GBM. In addition, the four targets, including Protection of Telomeres 1 (POT1), Adenylosuccinate Lyase (ADSL), FosB Proto-Oncogene, AP-1 Transcription Factor Subunit (FOSB), and Calcium Activated Nucleotidase 1 (CANT1), did not interact with other targets . Tumor P pathway ; pathway pathway . The GSE pathway .Uni-cox analysis revealed that 9 SG-TMZ-GBMs were determined as the significant survival-related risk genes. CCL5 was the most significant gene  . We founThe microenvironment cell population-counter method evaluated the association between CCL5 and 10 immune cell populations from transcriptomic data. A strong correlation between CCL5 and CD8 T cells, T cells, B lineage, and fibroblasts were seen . Then weSolanum nigrum L. (Panax ginseng, licorice, Lycium barbarum, Salvia miltiorrhiza bunge, Coptis rhizoma, and Sophora flavescens (GBM is the most frequent and the least treatable type of brain tumor, and the prognosis of GBM remains a challenge using the standard methods of treatment—TMZ, radiation, and surgical resection . TMZ is igrum L. , Panax gavescens . TCM is avescens . The antavescens .We tried to find a new TCM complementary method to treat GBM and hope that through combining Chinese and Western medicine, TMZ resistance could be reversed and anti-tumor therapeutic effects could be achieved. Luo Han Guo is a TCM with the same medicine and food. The multiple compounds in Luo Han Guo not only act on the same target protein, but a single compound also acts on various target proteins and multiple pathways, which reflects the “multiple components, multiple targets, and multiple pathways” of Luo Han Guo’s synergistic effect. Luo Han Guo may work with ar-turmerone, methyl palmitate, lauric acid, beta-sitosterol, gamma-aminobutyric acid, coumarin, mogroside V, and squalene. GO functional enrichment analyses reflected that most of the active ingredients in SG might target nerve cells.Through network pharmacology and bioinformatics analysis, we found that the CCL5 molecule is a potential target of SG, TMZ, and GBM, maybe the key to the clinical development of TMZ resistance . CCL5-CCCompared to genetically distinct syngeneic GBM models, the difference in mouse GBM models was eosinophils, reported in GBM . Eosinop


Conioselinum anthriscoides “Chuanxiong”  and Cyperus rotundus L.  (CRCR), is a classic prescription for treating migraine. This study aimed to reveal the potential mechanisms of CRCR extract against migraine using integrated analysis of metabolomics and network pharmacology. Behavioral changes in the nitroglycerin rat migraine model were determined from von Frey withdrawal response. Untargeted serum metabolomics was used to identify the differentially expressed metabolites and metabolic pathways. The differentially expressed metabolites were analyzed to obtain the corresponding targets by a compound–reaction–enzyme–gene network. Network pharmacology was used to construct a compound–target–pathway network. The common targets of metabolomics and network pharmacology were further analyzed. Metabolomics analysis identified 96 differentially expressed metabolites and 77 corresponding targets. Network pharmacology analysis identified 201 potential targets for CRCR against migraine. By intersecting 77 targets with 201 targets, monoamine oxidase A (MAO-A), monoamine oxidase B (MAO-B), and catechol-O-methyltransferase (COMT) were identified as the common targets, and MAO-A, MAO-B, and COMT were involved in the tyrosine metabolism pathway. Further experiments demonstrated that the contents of MAO-A and COMT were significantly increased in serum and brainstem tissue of the migraine rats. CRCR extract significantly decreased the contents of MAO-A and COMT, while no significant difference was found in MAO-B. Metabolomics analysis indicated that the contents of 3,4-dihydroxyphenylacetate (DOPAC) and 3-(4-hydroxyphenyl)pyruvate (HPP) were significantly increased in the migraine rats, and CRCR extract caused significant decreases in DOPAC and HPP. Interestingly, DOPAC and HPP were two differentially expressed metabolites involved in the tyrosine metabolism pathway. Correlation analysis showed that DOPAC and HPP were highly positively correlated with MAO-A and COMT. Taken together, two key differentially expressed metabolites (DOPAC and HPP), two key targets (MAO-A and COMT), and one relevant metabolic pathway (tyrosine metabolism) showed great importance in the treatment of migraine. This research could provide a new understanding of the potential mechanism of CRCR against migraine. More attentions should be paid into the tyrosine metabolism pathway in future studies.Migraine is a common neurological disorder that manifests as recurrent attacks of unilateral and throbbing headache.  Migraine is a chronic paroxysmal neurological disorder characterized by multiphase attacks of head pain and a myriad of neurological symptoms . The senConioselinum anthriscoides “Chuanxiong”  (known as Chuanxiong rhizoma in Chinese) is used to treat migraine and cerebral ischemic stroke. Cyperus rotundus L.  (known as Cyperi rhizoma in Chinese) is usually used to treat depression, epilepsy, and dysmenorrhea. Chuanxiong rhizoma and Cyperi rhizoma (abbreviated as CRCR), in a weight ratio of 1:2, is a classic prescription for treating migraine dating back to the Danxi Xinfa treatise in the Yuan Dynasty (AD 1347) of China. CRCR has the traditional clinical efficacy in activating blood circulation, dispelling wind, and relieving pain. Modern studies discovered analgesic, anti-inflammatory, antioxidant, anticerebral ischemia, and neuroprotective activities in CRCR , 3-n-butylphthalide (CAS 6066-49-5), senkyunolide A (CAS 63038-10-8), Z-ligustilide (CAS 81944-09-4), Z-3-butylidenephthalide (CAS 72917-31-8), cyperotundone (CAS 3466-15-7), nookatone (CAS 4674-50-4), and α-cyperone (CAS 473-08-5) were purchased from PUSH Bio-technology . All these chemicals were with a purity of greater than 98%. Nitroglycerin injections (NTGs) were obtained from Beijing Yimin Pharmaceutical Co., Ltd. . Sumatriptan succinate tablets were purchased from Tianjin Huajin Pharmaceutical Co., Ltd. . Rat 5-HT, CGRP, NO, and NOS ELISA kits were purchased from Nanjing Jiancheng Bioengineering Institute . Rat MAO-A, MAO-B, and COMT ELISA kits were purchased from Shanghai Enzyme-Linked Biotechnology Co., Ltd. . HPLC-grade methanol and acetonitrile were obtained from Fisher Scientific Inc. . Ultrapure water was produced by a Milli-Q Reagent Water System .Botanical drugs of Chuanxiong rhizoma (Batch No. 2001021) and Cyperi rhizoma (Batch No. D1908009) were obtained from Sichuan Neautus Traditional Chinese Co., Ltd. . They were identified by Dr. Li Ma  according to the 2020 edition of Chinese Pharmacopoeia. Voucher specimens were deposited in the Beijing Key Lab of Traditional Chinese Medicine Collateral Disease Theory Research of Capital Medical University.Chuanxiong rhizoma (62.5 g) and 125.0 g of Cyperi rhizoma were powdered, and then extracted by heating reflux with six volumes of 70% ethanol for 1 h and repeated twice. After filtered, the extraction solution was concentrated at 50°C using a vacuum rotary evaporator  and then dried in vacuum for the collection of dry extract. The yield of the dry extract was 20.71%.Eight main components were selected to be accurately quantified in the CRCR extract by HPLC according to our previous method . The conMale Sprague–Dawley rats (200 ± 20 g) were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. . Rats were housed in a temperature- and humidity-controlled environment. Rats were divided into four groups: control, NTG, sumatriptan, and CRCR groups. Rats were treated orally with saline at a dose of 10 ml/kg in the control and NTG groups, sumatriptan succinate tablets at a dose of 5.83 mg/kg in the sumatriptan group, and CRCR extract at a dose of 6.6 g/kg in the CRCR group for five consecutive days. After the last oral administration, rats were subcutaneously injected with 10 mg/kg of NTG to induce migraine attack. Two hours after NTG injection, the mechanical sensitivity thresholds were assessed by von Frey tests. Subsequently, rats were anesthetized by isoflurane inhalation. Blood was collected from the abdominal aorta and centrifuged at 4°C at 3,000 rpm for 15 min to obtain serum samples. Rats were sacrificed by cervical dislocation, and brainstem was separated on ice, and stored at −80°C until further analysis.Cutaneous allodynia is a typical symptom of migraine, with a crucial role in directing optimal treatment for migraine attacks . Cutaneov/v). After vortex mixing for 30 s and sonication for 30 min at 5°C, a sample was kept at −20°C for 30 min to improve protein precipitation. The sample was centrifuged at 13,000 ×g for 15 min at 4°C. The supernatant was evaporated at 37°C under a gentle nitrogen stream and then redissolved in 100 µl of ACN:H2O . After vortex mixing for 30 s and sonication for 5 min at 5°C, the sample was centrifuged at 13,000 ×g for 10 min at 4°C, and 2 µl of supernatant was subjected to UHPLC-Q-extractive MS analysis. For method validation, a pooled quality control (QC) sample was prepared by mixing an equal volume (20 µl) of each serum sample in parallel as above and inserted at regular intervals into the sequence.An aliquot of 100 µl of thawed serum sample was deproteinized with 400 µl of MeOH:ACN  and acetonitrile/isopropanol/water , both containing 0.1% formic acid. The flow rate was 0.4 ml/min. The elution gradient program was used as follows: 0–3.5 min, 100%–75.5% A; 3.5–5.0 min, 75.5–35% A; 5.0–5.5 min, 35%–0% A; 5.5–7.4 min, 0%–0% A; 7.4–7.6 min, 0%–48.5% A; 7.6–7.8 min, 48.5%–100% A; 7.8–10.0 min, 100%–100% A. The MS parameters were used as follows: spray voltage, ±3.5 KV; sheath gas flow rate, 50 arb; auxiliary gas flow rate, 13 arb; capillary temperature, 325°C; heater temperature, 425°C. The MS scan range was operated with the mass m/z 70–1,050 Da and with a 0.2-s accumulation time.Metabolomics analysis was performed by using a UHPLC system coupled with a Q Exactive hybrid quadrupole Orbitrap mass spectrometer . Chromatographic separation was performed by using an ACQUITY UPLC HSS T3 column  at 40°C. The mobile phase consisted of acetonitrile/water , METLIN (http://metlin.scripps.edu) and MycompoundID (http://www.mycompoundid.org). Metabolite enrichment analysis and pathway analysis were performed by Metaboanalyst 5.0 . The identified differential metabolites were used to construct a compound–reaction–enzyme–gene network in MetScape plugin in Cytoscape 3.7.1 software , retention time, and peak intensity. Multivariate statistical analysis was used to identify the differentially expressed metabolites by SIMCA-P 13.0 software . The differentially expressed metabolites were then identified based on the exact molecular weight and MS/MS spectrum based on online databases, e.g., Human Metabolome Database  and were manually replenished based on literature. Oral bioavailability (OB) and drug likeness (DL) were used to identify bioactive molecules. If a compound meets the criteria of OB ≥30% and DL ≥0.18, it will be chosen as a candidate molecule for further analysis. The SMILE structure of this molecule was imported into the Swiss Target Prediction database (http://www.swisstargetprediction.ch/) to obtain the corresponding targets. Furthermore, migraine-related gene targets were acquired from GeneCards (https://www.genecards.org/), Online Mendelian Inheritance in Man , DisGeNet (https://www.disgenet.org/), Therapeutic Target Database , and Drugbank (https://go.drugbank.com/).Compounds were retrieved from the Traditional Chinese Medicine Systems Pharmacology Database (https://www.uniprot.org/). Thus, a total of 1,801 migraine-related gene targets were retained by intersecting these targets from five different databases. The potential targets responsible for CRCR against migraine were acquired by intersecting CRCR-related gene targets with migraine-related gene targets. A protein–protein interaction network was constructed by STRING 11.0 (https://string-db.org/). The Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed by ClueGO plugin in Cytoscape 3.7.1 software.Migraine-related gene targets  were obtained from GeneCards database at first, and then 2,041 migraine-related gene targets were retained based on the median of the relevance score values. Migraine-related gene targets  were obtained from OMIM, DisGeNet, and TTD databases, respectively. From Drugbank database, 244 migraine-related protein targets were acquired at first, and 185 corresponding gene targets were obtained using Uniprot database .p-value of <0.05 was considered to be statistically significant. Pearson correlation analysis was performed using SPSS 17.0 software .Data were expressed as mean ± SD. Statistical analysis was performed by one-way ANOVA followed by multiple comparisons using GraphPad Prism 7.0 software . A p < 0.0001), and CRCR significantly elevated the mechanical threshold (p < 0.0001). p < 0.0001, p < 0.01, p < 0.0001, p < 0.0001). With the treatment of CRCR, 5-HT was significantly increased (p < 0.0001), CGRP and NOS were significantly decreased , which indicated CRCR had a good antimigraine effect on NTG-induced migraine rats.As shown in Pareto scaling and mean-centering pretreatment, a partial least squares discriminate analysis (PLS-DA) model was applied to identify the clustering property. As shown in R2 and Q2 values after 200 response permutation tests, indicating that the PLS-DA model was reliable with a low risk of overfitting.The metabolic profiles of serum samples were acquired by the UHPLC-Q-extractive MS data. After R2X = 0.487, R2Y = 0.992, Q2 = 0.647, R2X = 0.359, R2Y = 0.953, Q2 = 0.697, p < 0.05) were combined as the critical indicators to identify differentially expressed metabolites. A total of 1,897 metabolites were identified between the control and NTG groups, including 901 metabolites in the positive ion mode  was applied to analyze the overall metabolic distinction. A clear grouping trend between control and model samples could be observed in the positive ion mode  were acquired from GeneCards, OMIM, DisGeNet, TTD, and Drugbank databases. After intersecting 605 targets with 1,801 targets, 201 targets were identified as the potential targets for CRCR against migraine. A protein–protein interaction network was constructed by the Cytoscape software. nd so on . The topnd so on . FunctioO-methyltransferase (COMT). As shown in The identified differentially expressed metabolites were imported into Metscape to obtain a compound–reaction–enzyme-–gene network, which visualized the interaction among metabolites, pathways, enzymes, and genes. As shown in MAO-A, MAO-B, and COMT were identified as the common targets based on metabolomics and network pharmacology analysis. The contents of MAO-A, MAO-B, and COMT in rat serum and brainstem tissue were analyzed by ELISA. As shown in p < 0.0001, p < 0.0001, p < 0.05, p < 0.01), while the contents of phenylacetaldehyde, L-serine, L-tryptophan, and indole-3-acetamide were significantly decreased in the migraine rats . Treatment with CRCR caused significant decreases in the contents of DOPAC and HPP . However, the contents of phenylacetaldehyde, L-serine, L-glutamate, L-tryptophan, L-citrulline, and indole-3-acetamide were not statistically different between the NTG group and CRCR group. Thus, it can be speculated that DOPAC and HPP were the key endogenous metabolites for CRCR treating migraine.Correlation analysis was conducted between the peak areas of key endogenous metabolites (DOPAC and HPP) and the contents of key targets . The values of Pearson correlation coefficient are shown in The pathological mechanisms of migraine are very complicated, which have been associated with neurogenic inflammation, central sensitization, trigeminovascular damage, oxidative stress, and metabolic abnormalities . Recent 1-42-induced Alzheimer’s disease model mice with inhibitory effect against Aβ accumulation .Accumulating evidence has demonstrated that network pharmacology is an attractive tool to predict the interactions between multiple components and multiple targets in the botanical drugs . In receThe differentially expressed metabolites found in metabolomics were further analyzed to obtain the corresponding targets by a compound–reaction–enzyme–gene network. By intersecting 77 targets in the compound–reaction–enzyme–gene network with 201 targets in the protein–protein interaction network, MAO-A, MAO-B, and COMT were identified as the common targets. Interestingly, MAO-A, MAO-B, and COMT are the essential targets involved in the tyrosine metabolism pathway . A numbeIn the present study, DOPAC and HPP, the key metabolites involved in the tyrosine metabolism, were significantly increased in the migraine rats. MAO-A and COMT, the key enzymes involved in the tyrosine metabolism, were also significantly increased in the migraine rats. CRCR extract sharply reduced the contents of DOPAC and HPP, as well as MAO-A and COMT. Correlation analysis showed that DOPAC and HPP were highly positively correlated with MAO-A and COMT in the tyrosine metabolism pathway. Accordingly, it can be supposed that two key differentially expressed metabolites (DOPAC and HPP), two key targets (MAO-A and COMT), and one key metabolic pathway (tyrosine metabolism) were closely involved in the treatment of migraine by CRCR extract.The present study explored the potential mechanisms of CRCR for migraine treatment using integrated analysis of metabolomics and network pharmacology. The integrated analysis revealed that two key differentially expressed metabolites (DOPAC and HPP), two key targets (MAO-A and COMT), and one relevant metabolic pathway (tyrosine metabolism) showed great importance in CRCR treating migraine. This research could provide a new understanding of the potential mechanism of CRCR against migraine. More attention should be paid into the tyrosine metabolism pathway in future studies.


One of the critical issues hindering the commercialization of lithium–sulfur (Li–S) batteries is the dissolution and migration of soluble polysulfides in electrolyte, which is called the ‘shuttle effect’. To address this issue, previous studies have focused on separators featuring specific chemical affinities or physical confinement by porous coating materials. However, there have been no studies on the complex effects of the simultaneous presence of the internal and interparticle spaces of porous materials in Li–S batteries. In this report, the stable Zr-based metal–organic frameworks (MOFs), UiO-66, have been used as a separator coating material to provide interparticle space via size-controlled MOF particles and thermodynamic internal space via amine functionality. The abundant interparticle space promoted mass transport, resulting in enhanced cycling performance. However, when amine functionalized UiO-66 was employed as the separator coating material, the initial specific capacity and capacity retention of Li–S batteries were superior to those materials based on the interparticle effect. Therefore, it is concluded that the thermodynamic interaction inside internal space is more important for preventing polysulfide migration than spatial condensation of the interparticle space. To go beyond conventional lithium-ion batteries. Lithium–sulfur (Li–S) batteries have received much attention as one of the most promising candidates for a next-generation secondary battery systems because of their high theoretical energy density  have been applied as the separator coating materials and host materials owing to their distinct advantages, such as, a well-ordered porous matrix, high surface area, tunable morphology and pore structure, and amenability to functionalization ,23,24,252  for internal space control. The particle sizes of these MOFs were controlled to be large , medium , and small  for meticulous interparticle space control. These MOFs are referred to as UiO-66-H(X) and UiO-66-NH2(X) where X indicates the size of the MOF (2(X) series consistently exhibited higher specific capacity than those using the UiO-66-H(X) series, which is attributed to the strong adsorption of amine groups in UiO-66-NH2(X) towards soluble polysulfides. Consequently, the internal space effect of MOFs was found to be much stronger than their interparticle space effects towards the blocking of polysulfide migration in electrolytes.In this study, the effects of interparticle space have been compared with those of the internal spaces of MOFs in MOF-coated separators. Specially designed MOFs, featuring controlled particle sizes and functional groups, were used as a coating for separators in Li–S batteries, and their cycling performances were compared to determine the effect of their interparticle and internal spaces on polysulfide migration in Li–S batteries. UiO-66 was used as a representative MOF because of its high physical and chemical stability. The UiO-66s were functionalized with H and NH the MOF . After tPrevious studies using MOFs as separator coating materials have mostly focused on improving the cycling stability of batteries by introducing functional groups in the internal spaces ,29. AlthN,N-dimethylformamide , 1,3-dioxolane , 1,2-dimethoxyethane , lithium sulfide (99.98%), polyvinylidene Fluoride (PVDF) (MTI corporation), super-P 45 carbon (IMERYS), N-methyl-2-pyrrolidone (NMP), and bis(trifluoromethane)sulfonimide lithium salt  were purchased from Sigma-Aldrich Chemicals, Burlington, MA, USA. Zirconium (IV) chloride , acetic acid , and sulfur powder  were purchased from Alfa Aesar, Tewksbury, MA, USA. Carbon nanotubes  were purchased from Nanostructured & Amorphous Materials Inc, Los Alamos, NM, USA.Terephthalic acid (98%), 2-aminoterephthalic acid (99%), triethylamine , 2BDC, 0.15 mmol) (for UiO-66-H) or 2-aminoterephthalic acid  (for UiO-66-NH2) in DMF (5 mL). Separately, ZrCl4 (0.15 mmol) was dissolved in DMF (5 mL) and acetic acid (0.69 mL). This solution served as the metal source. The ligand and metal salt solutions were combined in a 20 mL glass vial. For synthesizing three distinctly sized UiO-66s, different quantities of TEA were added to the solutions as follows: 0 μL (for large size), 30 μL (for medium size), and 60 μL . The vial was placed directly in an aluminum heating block at 120 °C for 16 h. After the reaction, the vial was removed and cooled to room temperature. The powder product was separated using a centrifuge (6000 rpm for 10 min) followed by washing with DMF and methanol. After washing, the product was dried in a vacuum oven.Ligand solutions were prepared by dissolving terephthalic acid (H2) nanoparticles were mixed with PVDF binder in NMP at a mass ratio of 9:1 (UiO-66s:PVDF). The mixture was ground via ball-milling to obtain a homogeneous slurry. The slurry was coated onto Celgard 2400  using the doctor blade method. The coated separator was dried overnight in a vacuum oven at 60 °C. The obtained UiO-66s separator had approximately 20 μm thickness of MOF layer and was cut into a disk of diameter 16 mm.The synthesized UiO-66-H . The mass loading of sulfur in the electrode was ca. 0.6–1.0 mg/cm2. Coin cells  were assembled with lithium metal disks (15.6 mm diameter) as anodes and S/CNT as cathodes. The electrolyte was composed of 1 M LiTFSI dissolved in DOL/DME mixed solution  with 0.2 M LiNO3. The configuration of the assembled cell is represented as S/CNT cathode (12 mm)/UiO-66s coated separator (16 mm)/electrolyte (17 μL)/bare PP separator (18 mm)/electrolyte (17 μL)/Li metal anode (15.6 mm). The UiO-66s-coated layer was facing to the cathode side.Sulfur was ground with carbon nanotubes (CNTs) at a mass ratio of 7:3 to a homogeneous colored powder. Then, the powder mixture was heated from room temperature to 155 °C (holding for 12 h) and then through 260 °C (holding for 0.5 h) in a tube furnace under an argon atmosphere to enable the infiltration of sulfur into CNTs. The S/CNT composites were mixed with carbon black (Super-P 45 and PVDF binder in NMP at a mass ratio of 7:2:1 (S/CNT:Super-P:PVDF). The mixture was ground via ball milling and coated onto aluminum foil using the doctor blade method. Thereafter, the electrode was dried in a vacuum oven at 60 °C, punched to a diameter of 12mm and kept in an argon-filled glove box (HPowder X-ray diffraction (PXRD) spectra were obtained using a Bruker instrument (D8 Advance) at 1600 W . The scanning speed was 0.4 s/step at 0.04° increments. Nitrogen adsorption analysis was performed on a BELSORP-max automatic volumetric gas adsorption analyzer and the sample was activated by evacuating at 120 °C for 24 h. The morphology of MOF particles and coated separators was evaluated using a field emission scanning electron microscope . Attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopy was performed using a Nicolet iS50 FTIR spectrometer . The UV-visible absorption spectra were recorded using a Shimadzu UV-2600 instrument with quartz cuvettes.+) under the current densities of 100 mA/g and 250 mA/g. All specific capacities in this work were calculated based on the mass of sulfur. EIS experiments were performed in a frequency range from 1 MHz to 10 mHz using a potentiostat  with an AC oscillation amplitude of 10 mV.The galvanostatic charge/discharge measurements were performed using a BTS4008 battery cycler  between 1.8 to 2.8 V . In addition, the chemical environments of the pores can be conveniently modified by replacing the H in terephthalate with targeted functional groups such as −NH2, −OH, and −COOH.Zirconium-based MOFs are both chemically and electrochemically stable in battery systems ,33. In t2BDC), ZrCl4, and acetic acid were dissolved in 20 mL DMF in a Teflon-lined vial. To this solution, different doses of TEA were added to obtain UiO-66s of varying particle sizes. TEA was used to deprotonate the carboxyl groups of organic linkers and ensure dominant nucleation to produce UiO-66s of small particle sizes. The larger amount of TEA used, the smaller the UiO-66 particle size. This mixture was sealed and placed at 120 °C for 16 h in an aluminum heating block. The heating block has a high heat transfer rate, which is advantageous for synthesizing uniform nano-sized MOFs. The white suspensions were collected and washed with DMF and methanol using a centrifuge. The obtained MOFs were then immersed in methanol for three repeated 24 h periods to activate their pores. The UiO-66-H(X) particles were separated and dried under vacuum overnight. The same synthetic procedure was followed to obtain UiO-66-NH2(X) particles except that the ligand reagent was changed to 2-aminoterephthalic acid. All products were characterized using PXRD, SEM, FT-IR, and N2 adsorption isotherms measurements to determine the intrinsic crystallinity, morphology, functionality, and permanent porosity of UiO-66s particles, respectively, which will be discussed in the following sections.The UiO-66s were prepared via a typical solvothermal synthesis that was modified from a reported method  to contr2(X) samples matched well with those simulated results from their crystal structures, indicating that they possess the same UiO-66 crystal structure (2 adsorption measurements as shown in 2(X) species were similar in micropore filling regions at low relative pressure. However, both the UiO-66-H(S) and UiO-66-NH2(S) showed hysteresis at high relative pressures (P/P0) above 0.6, which was different from the behaviors of UiO-66-H(L) and UiO-66-NH2(L). This is because UiO-66-H(S) and UiO-66-NH2(S) had small interparticle spaces corresponding to those of mesoscale pores (2(L) had large interparticle spaces that were beyond the scope of N2 adsorption measurements. SEM images depict the high uniformity of UiO-66-H(X) and UiO-66-NH2(X) structures along with octahedral geometry that is typical of UiO-66. As shown in 2(X) samples have large (L), medium (M), and small (S) particle sizes of ca. 400 nm, 100 nm, and 20 nm, respectively. Consistent with previous PXRD results, UiO-66-H(X) and UiO-66-NH2(X) samples show sharp edges in their octahedral shapes indicating high crystallinity. These results confirmed that the prepared crystals of UiO-66-H(X) and UiO-66-NH2(X) had the desired particle sizes and interparticle spaces.The PXRD patterns of both UiO-66-H(X) and UiO-66-NHtructure . As the le pores b. In con2Sn, 4 < n ≤ 8) are soluble in electrolyte, it is important to block them in the cathode side by the coating materials on the separator to further mitigate the shuttle effect in Li–S batteries. Absorption experiments of polysulfides were performed by immersing UiO-66-H(X) and UiO-66-NH2(X) samples in Li2S6  solution. The Li2S6 solution was prepared according to a previously reported method [2S6 solution faded instantly and became colorless. It was kept on observation for 10 h, while the UiO-66 powder turned from a white to yellow color  samples, the absorption ability was obviously affected by particle size: the absorbance at 320 nm decreased gradually with a decrease in particle size. On the other hand, as for the UiO-66-NH2(X) species, no significant differences in absorbance were observed with different particle sizes. Moreover, the absorbance of UiO-66-NH2(X) samples was much lower than that of UiO-66-H(X) samples. These results indicate that the amine functionality in UiO-66-NH2(X) imparts a strong affinity towards polysulfides, overwhelming the differences in their particle sizes.As polysulfides  samples showed identical FT-IR patterns regardless of their particle sizes. As shown in 2S6 immersion treatment, two new peaks appeared at 2985 and 2890 cm−1 for all UiO-66-H(X) samples, suggesting the absorption of Li2S6 (2(X) species showed identical FT-IR patterns regardless of their particle sizes and the presence of Li2S6 peaks appeared at 2989 and 2892 cm−1  samples, the peak corresponding to C–N stretching at 1254 cm−1 was shifted to 1258 cm−1. Additionally, the intensity of the peak at 3469 cm−1 that was assigned to N–H stretching increased after Li2S6 absorption treatment  samples could absorb polysulfides by their micropores like a molecular sieve. While the particle size has a distinct influence on the polysulfides absorption ability of UiO-66s, the internal space interactions derived from amine groups had much stronger effects. Therefore, the UiO-66-NH2(X) samples exhibited much more effective polysulfide absorption than UiO-66-H(X) sampled in all size ranges.In conjunction with the UV-vis and FT-IR results, UiO-66-H(X) and UiO-66-NH2(X) coated separators (diameter: 16 mm) exhibited a uniform distribution of MOF particles. The interparticle spaces for each separator are represented by the black regions in the contrast images (2(X) ones. Moreover, as the void space decreased, the coated layer became dense when the particle size was small, which is consistent with the N2 adsorption analysis of UiO-66-H(X) and UiO-66-NH2(X)  b. It was2(X)) coated on PP membrane), a lithium metal disk as the anode and 1 M LiTFSI-DOL/DME  with 0.2 M LiNO3 as the electrolyte. All cells were tested under the current density of 250 mA g−1 in the voltage range of 1.8–2.8 V (vs. Li/Li+) after the activation step at 100 mA g−1 for the first cycle (2(X) cells. During discharging, elemental sulfur was reduced to soluble Li2Sn (4 < n ≤ 8) and finally insoluble Li2S2/Li2S with two main voltage plateaus at 2.3 V and 2.1 V (vs. Li/Li+) [2S2/Li2S were oxidized back to S8 via the soluble polysulfides intermediates. These behaviors showed that both UiO-66-H(X) and UiO-66-NH2(X) had no electrochemical influence on the typical charge/discharge redox process in the Li–S battery. Nevertheless, cells with UiO-66-NH2(X) separators exhibited higher specific capacities than those with UiO-66-H(X) ones, indicating the enhanced utilization of sulfur because of the strong adsorption ability of amine groups towards polysulfides as discussed from UV-vis results in The fabricated MOF-coated separators were employed in Li–S batteries to investigate the internal and interparticle space effects on the polysulfide migration. The CR2032 type coin cells were assembled with S/CNT as the cathode  and UiO-66-NHst cycle . As show Li/Li+) ; during −1 and final: 271.20 mAh g−1) than UiO-66-H(M) and UiO-66-H(S) with capacity retention values of 42.2%  and 29.2% , respectively, for the 350th cycle. The cross point of capacity fading appeared after 75 cycles depending on the particle size. Smaller particles are coated on the PP separators less reliably than larger particles having larger interparticle spaces. We also summarized the previously reported UiO-66 coated separators in Li–S batteries in 2. In this work, we further studied the effects of internal and interparticle space on polysulfide migration via size control and amine group modification, which could fill the gap between the microstructure of UiO-66s and their electrochemical application. To understand the effect of particle size on the cyclability of UiO-66-H(X) cells, EIS were conducted for investigating the reaction governing the conversion of soluble polysulfide to insoluble Li2S over three different frequency regions: high-frequency region, middle-frequency region, and low-frequency region. In the high-frequency region , the series ohmic resistances (Rs) of the electrolyte, soluble polysulfide, and electrodes dominate over their diffusion. In the middle-frequency region, charge transfer resistance (Rct) reveals electronic resistance at the solid-liquid interfaces. In the low-frequency region, Warburg impedance (Zw) is observed, that is, diffusion dominates over the ohmic resistance. Therefore, each frequency region provides insight into the properties of the resistance and the diffusion as a function of the degree of interparticle spaces. In the Nyquist plot , UiO-66-H(M), and UiO-66-H(S) cells were estimated to be 6.14 Ω and 56.3 Ω, 2.474 Ω and 43.21 Ω, and 2.585 Ω and 38.06 Ω, respectively. The Rs and Rct values of UiO-66-H(S) are smaller than those of UiO-66-H(M) and UiO-66-H(L) (s and Rct of UiO-66-H(S).Although there was no noticeable polarization difference among the UiO-66-H(X) species, significant differences were observed in the long-term cycling performances c. UiO-66ist plot d, Rs, Rcit model . From th-66-H(L) , indicatw values exhibit the opposite tendency of becoming reduced with increasing interparticle space fractions. In the low-frequency region, Zw values were strongly related to diffusion via MOF particles within the separator, indicating that the diffusion of soluble polysulfide can be regulated by controlling the balance between internal and interparticle spaces. The Zw values of UiO-66-H(L), UiO-66-H(M), and UiO-66-H(S) are 14.76 Ω s−1/2, 16.28 Ω s−1/2, and 21.35 Ω s−1/2, respectively. Smaller Zw values signify faster diffusion of ions, such as polysulfide and lithium ion. Thus, these results imply that diffusion depends on the interparticle spaces, although UiO-66-H(S) exhibited better adsorption of Li2S6 than UiO-66-H(M) and UiO-66-H(L). The Nyquist plots of the 350th cycles were fitted to reaffirm the diffusion tendency of UiO-66-H(L) using a modified equivalent circuit model  was 6.906 Ω s−1/2, which is much smaller than that of UiO-66-H(M) (9.265 Ω s−1/2) and UiO-66-H(S) (29.93 Ω s−1/2)  and UiO-66-H(M) decreased after 350 cycles, implying that the diffusion was accelerated by the interparticle spaces after repeated charging and discharging. As for UiO-66-H(S) cells, the Zw value increased conspicuously after 350 cycles, suggesting that the insufficient interparticle spaces hindered diffusion during cycling because of restricted mass transport channels. Therefore, densely packed particles did not effectively prevent the shuttle effect of polysulfides due to the limited diffusion into the micropores of UiO-66-H(X). The passing soluble Li2Sn (4 < n ≤ 8) resulted in the irreversible formation of insoluble Li2S2/Li2S on lithium anode as shown in the capacity degradation. On the contrary, large interparticle spaces of UiO-66-H(L) promoted ion diffusion despite an increase in electric resistance, leading to good capacity retention after the 350th cycle.On the other hand, Zit model . After 3Ω s−1/2) . The Zw 2(X) and UiO-66-H(X) were compared. As demonstrated in 2(X) cells are much higher than those of UiO-66-H(X) ones. Moreover, there is no significant capacity difference among the UiO-66-NH2(X) species. This was because the amine functional groups had a significantly larger preventive influence on the shuttle effect of soluble polysulfides than the interparticle space effects. In the initial reduction step of elemental sulfur, soluble polysulfide intermediates were trapped on the coated separators. The EIS fitting parameter of UiO-66-H(X) demonstrated that the existence of sufficient interparticle spaces improved the diffusion of polysulfide intermediates. Additionally, the amine group strengthened the Van der Waals force between the trapped polysulfides and UiO-66-NH2(X) via the unpaired-electrons of nitrogen, thus making them more polar. According to the literature on nitrogen effect, the incorporation of nitrogen enhanced ion mobility by providing a thermodynamically favored route for diffusion [2S6 was carried out (2S6 solutions containing UiO-66-NH2(X) coated separators especially UiO-66-NH2(L) are brighter than those containing UiO-66-H(X) ones. The internal space effect originating from the enhanced absorption of the amine group is strong enough to suppress polysulfide migration through separators regardless of the interparticle space effects, that is, the difference in void spaces. This result also proved that the effect of internal space on the chemical affinity surpassed the effect of interparticle space arising from particle size control.Subsequently, the internal space effects in UiO-66-NHiffusion . To veriried out . The col2(X) separators were much improved by the amine groups regardless of the difference in interparticle spaces. While many studies have been conducted on the performance improvement of separators using MOFs with different structures, pore sizes, metal clusters, and organic ligands, in this study, we determined that the internal space effect is more critical to mitigating polysulfide migration than interparticle space effects when other factors that could potentially cause side effects were carefully controlled. We expect that the findings in this study will contribute to the optimized design and manufacturing of MOF-modified separators in other battery systems such as sodium–sulfur batteries.Through this study, we discovered the key factors in preventing polysulfides migration in Li–S batteries. By using UiO-66s as coating materials in which the pore structure and framework are equally controlled, separators were designed with three different UiO-66 particle sizes  by controlling the balance between their internal and interparticle spaces. Furthermore, amine functional groups were introduced into UiO-66s to enhance the absorption ability towards polysulfides. We found that Li–S cells with UiO-66-H(x) separators exhibited distinct cycling performances, which greatly depends on the particle sizes and interparticle space of UiO-66-H(X). However, this interparticle space effect became inconspicuous when the internal space caused by amine groups was the dominated effect on the cyclability of Li–S cells. Therefore, the specific capacity and cycling performance of Li–S cells with UiO-66-NH


Chronic rhinosinusitis (CRS) is a complex condition brought on for many reasons, and its prevalence is rising gradually around the world. Xanthii Fructus (XF) has been used in the treatment of CRS for decades and is effective. The chemical and pharmacological profiles of XF, on the other hand, are still unknown and need to be clarified. The potential mechanisms of XF in CRS treatment were investigated using a network pharmacology approach in this study. OB and DL were in charge of screening the bioactive components in XF and drug-likeness. TCMSP and PubChem databases were used to identify prospective XF proteins, whereas GeneCards and the DisGeNET database were used to identify potential CRS genes. An interactive network of XF and CRS is built using the STRING database based on common goals identified by the online tool Venny. Cytoscape was used to visualize the topological characteristics of nodes, while the biological function pathways were identified by GO Knowledge Base, KEGG. There were 26 bioactive components and 115 potential targets in XF that bind to CRS or are considered therapeutically relevant. Five significant signaling pathways have been found for CRS by the pathway analysis including the HIF-1 signaling pathway, TNF signaling pathway, Toll-like receptor signaling pathway, NOD-like receptor signaling pathway, and PI3K-Akt signaling pathway. We simultaneously confirmed that the PI3K-Akt pathway promotes the development of CRS. Finally, this study took a holistic approach to the pharmacological actions and molecular mechanisms of XF in the treatment of CRS. TNF, INS, CCL2, CXCL8, IL-10, VEGFA, and IL-6 have all been identified as potential targets for anti-inflammatory and immune-boosting effects. This network pharmacology prediction could be useful in manifesting the molecular mechanisms of the Chinese herbal compound XF for CRS. Chronic rhinosinusitis (CRS) is an inflammation of the nasal and sinus, which has a complex etiology including immunological and epithelial barrier components affected by the microbiota, environment, and genetic factors, while the course of CRS is more than 12 weeks , while 1Xanthium sibiricum Patrin is called Xanthii Fructus (XF). It' has been used as an herbal medicine for thousands of years to treat quite a few diseases, such as nasal blockage, sinusitis with nasal discharge, nasal discharge, urticaria with itching, and rhinitis with muscle spasms caused by wind and moisture [People are progressively pursuing health, while the notion of healthy living changed dramatically. Traditional Chinese medicine's (TCM)' distinctive effects not only evoked the interest of domestic intellectuals but also had stirred up the interest of the worldwide community . The frumoisture . Huang emoisture  found thmoisture  discovermoisture  discoverIn this article, the studies related to XF for the treatment of CRS were carried out by the network pharmacology approach. The target proteins were selected among the ingredients of XF and the treatment of CRS, and the core was built by compound-gene-disease (C-G-D) network between XF and CRS. Using overlapping genes, protein-protein interaction (PPI) networks were created. Hub genes were chosen from Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. The goal of this research was to find a possible therapeutic target for XF in the treatment of CRS and to create the groundwork for further research into the pharmacological mechanism of XF see .https://tcmsp-e.com/) was used to find all of the chemicals in XF [The traditional Chinese medicine systems pharmacology (TCMSP) database (ls in XF . The mosls in XF . The rathttp://www.tcmspw.com/tcmsp.php) and the PubChem database (https://pubchem.ncbi.nlm.nih.gov/) [http://www.uniprot.org/) [Compounds influence biological activity at both the transcriptional and protein levels by binding to certain molecular targets. The chemical-target interaction was necessary for elucidating the mode of compound action, which was obtained from the TCMSP database (ih.gov/) . The Pubot.org/) . The tarhttps://www.genecards.org/) and DisGeNET database (http://www.disgenet.org/) using the keyword “chronic rhinosinusitis.” The GeneCards database contains detailed information on human genes, which have been annotated [http://bioinfogp.cnb.csic.es/tools/venny/), the intersection between the probable target genes of XF and CRS-related targets was determined, and these were considered potential targets in XF for the treatment of CRS, which was visualized with a Venn diagram.To discover CRS-related targets, we searched the GeneCards database (nnotated . The Disnnotated . With thhttps://cytoscape.org/) [To get the overlapping genes, we took the target genes of XF and the therapeutic targets of CRS from the above four databases. Following that, we combined the information from the component, genetic, and diseases while we construct C-G-D using the Cytoscape software  .https://string-db.org/, version 11.0) [For PPI network research, the common genes of XF and CRS have been uploaded to the STRING database. PPI network visualization was established on associated proteins with a confidence score ≥0.4 (on 11.0) . In the on 11.0) . The nethttps://david.ncifcrf.gov/), a collection of gene annotation and analysis resources [R chooses the top 10 BP, CC, and MF GO keywords, as well as the top 20 KEGG pathways, bubble diagrams for functional annotation clustering to structure enrichment analysis, based on a high count and p < 0.05 [GO Knowledge Base can help us learn about gene functions such as acellular components (CCs), biological processes (BPs), and molecular functions (MFs) , and KEGesources . The clup < 0.05 . Enrichmβ-actin were acquired from Abcam, while secondary antibodies labeled with horseradish peroxidase were purchased from Shanghai Biyuntian Co.Thirty healthy rabbits at the age of 3 months, weighing 1.6-2.5 kg, were purchased from Fengdong New City Experimental Animal Farm, Xi'an New District (Certificate No. (SCXK(Shan)2017-002). Animals were kept in the SPF animal laboratory at room temperature of 20-25°C, relative humidity of 45%-50%, normal light, and free access to food and water. The maker of XF granules was Guangdong Yifang Traditional Chinese Medicine Granules Co., Ltd., which was obtained from Hainan Provincial Hospital of Traditional Chinese Medicine. The experimental procedure adheres to applicable animal ethics guidelines. 2) Primary antibodies against PI3K, AKT, and Staphylococcus aureus was inoculated in the culture medium and placed in an incubator for 24 h. Colonies were collected and diluted with saline into a bacterial suspension of 1 ms unit and set aside.Bacteria were obtained from maxillary sinus puncture rinses of outpatients with CRS, and a single strain of Staphylococcus aureus suspension was injected, the surgical wound was disinfected with iodophor, and the periosteum and skin were sutured layer by layer. The surgical procedure was the same for the XF group. The normal control group did not require any treatment. The duration of modeling was 42 d. From the seventh week, the XF group was treated with 350 mg/(Kg.d) of gavage, and the rest of the groups were treated with an equal volume of saline, and the duration of treatment for each group was 14 d. Afterward, the nasal tissue was removed by execution.Thirty rabbits were divided into three groups at random: the control group, the model, and the XF group, with 10 rabbits in each group. A longitudinal incision was made along the midline of the nasal dorsum, and the subcutaneous tissue and periosteum of the anterior maxillary sinuses were separated bilaterally. The maxillary sinus puncture needle was used to drill a small hole of 1.5 mm in diameter in the depression of the round hole of the anterior maxillary sinus wall; meanwhile, a few cotton wool was placed in the sinus opening and sinus cavity. Then, 1 mL (3∗108(CFU)/mL) of J software after the Western blot bands were evaluated.Nasal tissues were extracted in each group, and extracted total protein was used to detect PI3K and AKT expression after grinding and milling. The concentration of each set of proteins was determined using the BCA technique, and the proteins were separated using SDS-PAGE electrophoresis. The proteins were transferred to the PVDF membrane, followed by 5% skim milk closure for 2 h. Anti-rabbit PI3K protein (1 : 1000) antibody and anti-rabbit AKT protein (1 : 2000) antibody were added, kept warm at 4 °C, and incubated overnight, the primary antibody was washed away by TBST, secondary antibody (1 : 5000) was added at room temperature 1 h, TBST washed off the secondary antibody and imaged with ECL chemiluminescent solution for 3 min, and the target bands/corresponding relative expression levels were calculated using Imaget-test with p < 0.05.Based on the characteristics of comparison, the statistical analyses were conducted with Excel 2016 and GraphPad Prism version 5.0 software, using an unpaired Based on the OB and DL values, the TCMSP database was searched to locate 26 active components. The active compounds were supplanted by numbers to facilitate future data analysis see .The TCMSP database and the PubChem database yielded 1603 targets among the 26 candidate components, after removing the overlapping target, and there were a total of 638 ingredient targets in XF. Since removing duplicates from the GeneCards and DisGeNET databases, 855 genes involved with CRS were discovered. After that, using the Venny 2.1, the intersection of 638 XF targets and 855 CRS-related targets was determined. As a consequence, Venn diagrams revealed 115 potential genes coupled to both CRS and XF see .Because Chinese medicinal formulae have multiple pharmacological impacts by interacting with various targets, network analysis is a useful method for studying the underlying mechanisms of Chinese medicinal formulae on complicated disorders. The construction C-G-D network of XF on CRS was erected using Cytoscape 3.6.0 see . The netIncreasing evidence suggests that diseases are caused by interactions among multiple targets rather than by a single gene. Potential genes were entered into the STRING database to build a PPI network to further elaborate the molecular mechanisms of XF's pharmacological actions on CRS. Proteins and protein-protein interactions are represented by the network's nodes and edges. After hiding disconnected nodes, the network was established with 115 nodes and 2041 edges in this study see . The aveThe above common targets were entered into DAVID for GO enrichment analysis to show the numerous biological functions of probable targets in CRS following XF therapy. The biological process (BP) was significantly enriched in the cellular response to interleukin-1 (GO:0071347), cellular response to organic cyclic compound (GO:0071407), lipopolysaccharide-mediated signaling pathway (GO:0031663), positive regulation of pri-miRNA transcription from RNA polymerase II promoter (GO:1902895), and cellular response to tumor necrosis (GO:0071407), cellular components (CCs) in membrane raft (GO:0045121), cytosol (GO:0005829), apical plasma membrane (GO:0016324), focal adhesion (GO:0005925), and cytoplasm (GO:0005737), molecular function (MF) in protein tyrosine kinase activity (GO:0004713), Ras guanyl nucleotide exchange factor activity (GO:0005088), phosphatidylinositol-4,5-bisphosphate 3-kinase activity (GO:0046934), transcription regulatory region DNA binding (GO:0044212), and transcription factor binding (GO:0008134) see .P values  levels and myeloperoxidase (MPO) activity. Furthermore, in a dose-dependent way, carrageenan-induced production of several pro-inflammatory cytokines, such as TNF, IL-1, IL-6, PGE2, and NO, was decreased in mice pretreated with β-patchoulene [β-sitosterol. β-sitosterol inhibited caspase-1 activation and decreased the expression of NLRP3, a key component of the NLRP3 inflammasome. NF–B was partially suppressed in macrophages [According to the literature, anatomical blockage of the stomatal complicated, impaired respiratory system, chronic sinusitis, microbes, biofilms, immune disorder, deficient epithelial defense, biological factors, and environmental factors are all factors that influence the prognosis of CRS . Linoleichoulene . Inflammrophages . By blocrophages . With alα is a kind of TNF. In CRS, TNF-α causes small proline-rich protein dysregulation. This may have resulted in a breakdown of the epithelial barrier. VEGFA can promote tissue remodeling and angiogenesis in CRS. CCL2 is a chemokine that plays a pivotal role in the inflammatory response. It acts as a chemoattractant for monocytes and macrophages and is involved in a variety of immune processes [Several genes implicated in the occurrence and development of CRS were found among some of the top 10 hub targets, and they play a critical role in the therapeutic value of XF for CRS treatment. In CRS, the pro-inflammatory effects of IL-6 trans-signaling have been connected to the transition from acute innate immune responses to adaptive chronic inflammatory reactions . IL-6 inrocesses . CXCL8 arocesses . By attarocesses . Rudack rocesses . In CRS,rocesses . INS canrocesses . Based oκB is discharged from its inhibitors by RIPK2, which activates a kinase complex. NF-κBIA is the most well-studied NF-κB inhibitor, and it works by sequestering NF-κB in the cytoplasm and blocking it from binding to DNA, therefore inhibiting NF–B-dependent transcription. Nod2 activation in CRS causes the breakdown of NF-κBIA and subsequent release of NF-κB, which translocates into the nucleus to transcribe pro-inflammatory genes [As shown in a KEGG pathway enrichment analysis, many signaling pathways were utilized in the care of CRS with XF in this study. XF-associated hub targets were found in the HIF-1 signaling pathway, TNF signaling pathway, Toll-like receptor signaling pathway, NOD-like receptor signaling pathway, and PI3K-Akt signaling pathway, which are all closely relevant to CRS. The HIF-1 signaling pathway is a multifunctional angiogenic regulator that promotes endothelial cell proliferation and migration, while also increasing vascular permeability , plasma ry genes . When cory genes , which cry genes . A largeStaphylococcus aureus and Streptococcus viridans) and Gram-negative enterobacteria have been found. Mucosal inflammation with neutrophil and monocyte infiltration, extracellular matrix reconstruction, cupped cell proliferation, and tissue necrosis are all symptoms of bacterial infection [In rhinology, CRS is a prevalent condition, with chronic sinusitis without nasal polyps (CRSsNP) accounting for roughly 60% of cases. CRSsNP typically begins as a bacterial infection that progresses to chronic inflammation if left untreated . In the nfection , 54. Fornfection  found thnfection . As a reThis study, however, had some limitations. For example, it was unclear whether the pathways studied in this study were downregulated or upregulated. At the same time, we focus on the animal model of CRSsNP and use this model to demonstrate that XF has a regulatory role in CRSsNP through the PI3K/AKT pathway. However, the pathogenesis of chronic rhinosinusitis with nasal polyps (CRSwNP) is somewhat different from that of CRSsNP, so this model cannot be used to show the same role in CRSwNP. In addition, the experimental validation of this study was mainly at the animal level and did not involve human specimens, and it remains to be studied whether XF plays the same role in the treatment of CRS in humans as it does in animal studies. The network pharmacology approach identified potential targets of XF in CRS treatment, but these are theoretical predictions for now, which need to be confirmed in clinical and cellular or animal studies.