Limd1 gene undergoes mutation only at a low rate in NSCLC (0.712%). We have further identified LIMD1-associated molecular signatures in NSCLC, including its natural antisense long non-coding RNA LIMD1-AS1 and a pool of membrane trafficking regulators. We have also identified a subgroup of tumor-infiltrating lymphocytes, especially neutrophils, whose tumor infiltration levels significantly correlate with LIMD1 level in both LUAD and LUSC. However, a significant correlation of LIMD1 with a subset of immune regulatory molecules, such as IL6R and TAP1, was only found in LUAD. Regarding the clinical outcomes, LIMD1 expression level only significantly correlates with the survival of LUAD (p<0.01) but not with that of LUSC (p>0.1) patients. These findings indicate that LIMD1 plays a survival role in LUAD patients at least by acting as an immune regulatory protein. To further understand the mechanisms underlying the tumor-suppressing function of LIMD1 in NSCLC, we show that LIMD1 downregulation remarkably correlates with the deregulation of multiple pathways that play decisive roles in the oncogenesis of NSCLC, especially those mediated by EGFR, KRAS, PIK3CA, Keap1, and p63, in both LUAD and LUSC, and those mediated by p53 and CDKN2A only in LUAD. This study has disclosed that LIMD1 can serve as a survival prognostic marker for LUAD patients and provides mechanistic insights into the interaction of LIMD1 with NSCLC, which provide valuable information for clinical applications.Non-small-cell lung carcinoma (NSCLC) is the major type of lung cancer, which is among the leading causes of cancer-related deaths worldwide. LIMD1 was previously identified as a tumor suppressor in lung cancer, but their detailed interaction in this setting remains unclear. In this study, we have carried out multiple genome-wide bioinformatic analyses for a comprehensive understanding of LIMD1 in NSCLC, using various online algorithm platforms that have been built for mega databases derived from both clinical and cell line samples. Our results indicate that LIMD1 expression level is significantly downregulated at both mRNA and protein levels in both lung adenocarcinoma (LUAD) and lung squamous cell carcinoma (LUSC), with a considerable contribution from its promoter methylation rather than its gene mutations. The  Lung cancer is among the leading causes of cancer-related deaths in the world. There are nearly 230 thousand of new lung cancer patients and more than 135 thousand of deaths in 2020 in the USA (American Cancer Society). The 5-year relative survival rate of lung cancer from 1995 to 2001 was 15.7%. Non-small-cell lung carcinoma (NSCLC) is the major histological type of lung cancer that contributes to approximately 85% of all lung cancers. Like small-cell lung carcinoma (SCLC), NSCLC is originated from epithelial cells, and includes two main subtypes: lung adenocarcinoma (LUAD) and lung squamous cell carcinoma (LUSC). NSCLC patients are relatively insensitive to chemotherapy and radiation therapy compared with SCLC patients, and innovative therapeutic strategies are desired.Limd1 gene is located at chromosome 3p21.3 , and molecular and immune signatures in this setting. More importantly, we found that LIMD1 significantly correlates with the survival of LUAD but not that of LUSC patients.We employed different online algorithms for metadata analysis, including Oncomine, Genotype-Tissue Expression (GTEx), Gene Expression Atlas, ProteinAtlas, ProteomicsDB, Tumor Immune Estimation Resource (TIMER v2) , 16, GenAll analyses were carried out using the default settings of the corresponding algorithms if not otherwise indicated, with the detailed dataset information and guidelines available on each algorithm portal. p<0.05 is considered statistically significant and >0.05 is non-significant (n.s), and p<0.01 is considered statistically very significant.Limd1 gene produces four alternatively spliced variants (Limd1 transcription (data not shown). Consistent with these results, our recent studies have shown that LIMD1 is induced by NFκB and IRF4 downstream of the signaling pathway triggered by the EBV principal oncoprotein LMP1 in EBV-transformed LCLs, suggesting that LIMD1 could serve as an oncogenic biomarker in association with NFκB and IRF4 in certain hematological malignancies , functional annotation of the mammalian genome (FANTOM v5), and GTEx projects. The gnancies , 12.Regarding cell-specific expression, analysis in the ProteinAtlas portal shows that the transcript abundance of LIMD1 is mainly enriched in the cells originated from lymphoid, brain, and lung , uterine corpus endometrial carcinoma (UCEC), and uterine carcinosarcoma (UCS), and upregulated in lymphoid neoplasm diffuse large B-cell lymphoma (DLBC), breast cancer (BRCA), glioblastoma multiforme (GBM), acute myeloid leukemia (LAML), and brain lower-grade glioma (LGG) 2. In agLimd1 gene promoter is highly methylated in a region spanning nucleotides 45559963 to 45678725 in both LUAD and LUSC, exemplified by the probe cg15765353 that covers the two nucleotides 45559963-4 (p=4e-7 for LUAD and p=9.1e-11 for LUSC) , but undergoes higher rates in DLBC (2/37), UCEC (16/531), and cholangiocarcinoma  , with LUSC (>75%) being higher than LUAD (~50%)  were detected in 33 out of 226 lung cancer samples (14.6%), with most being missense substitution (57.58%) (Limd1 has a rate of 0.712% of mutations in NSCLC patients (43/6040) (not shown). ChimerDB and TumorFusions portals indicate that, among these detected Limd1 mutation types, Limd1-Lars2 gene fusion occurs in both LUAD and LUAC tumors and also in STAD, with Tier 1 in-frame fusion in LUAD and Tier 2 out-of-frame fusion in LUSC  , TCGA luTogether, these findings support the claim that the deregulation of LIMD1 expression in different cancers mainly results from epigenetic reprogramming rather than its mutation, with its downregulation in NSCLC being partially attributed to promoter methylation and arm-level deletion. In addition to these epigenetic mechanisms, miRNAs likely regulate LIMD1 expression in different cancer contexts, with miR-127-5p, miR-147a, and miR-130b-3p that potentially target LIMD1 in LUAD, as analyzed in DriverDBv3 , in the Oncomine portal. Results show that LIMD1 downregulation occurs at the early stage of cancer development in both LUAD and LUSC, and there are no significant differences between every two stages  to profile LIMD1-associated molecular signatures in lung cancer, in Oncomine, GEPIA2, and UALCAN portals, using the Pearson correlation test. These analyses identified a panel of common genes that correlate with LIMD1 (PCC>0.5) at the mRNA level in both LUAD and LUSC, and representative results from the GEPIA2 portal are shown in LIMD1-AS1 perfectly correlates with LIMD1 not only in lung cancer, but also in all other types of cancers in the TCGA dataset . RBSN beAdditionally, we identified LUAD- and LUSC-specific genes that correlate with LIMD1 at the transcriptional level in GEPIA2 and UALCAN portals (Tumor-infiltrating lymphocytes (TILs), an important component of the tumor microenvironment (TME), can recognize and kill cancer cells, and are an ideal candidate for cellular immunotherapy for various cancers –46. EspeWe, therefore, sought to assess the correlation of LIMD1 expression with different subtypes of TILs in NSCLC, including CD4 T cells, CD8 T cells, B cells, NK cells, dendritic cells (DCs), macrophages, myeloid-derived suppressor cells (MDSCs), mast cells, neutrophil, eosinophils, cancer-associated fibroblasts, endothelial cells, common lymphoid and myeloid progenitors, and hematopoietic stem cells, using Spearman correlation test in TIMER2 portal. We set the cutoff values of the Spearman coefficient to R<-0.33 (negative correlation) or R>0.33 (positive correlation), and p<0.05 was considered significant. Within these cutoff criteria, we found that LIMD1 expression levels positively correlate with the infiltration of neutrophils, CD4+ T cells, endothelial cells, and mast cells in both LUAD and LUSC . FurtherLimd1 gene promoter methylation levels significantly correlate with the frequencies of activated tumor-infiltrating CD4+ and CD8+ T cells and B cells (R<-0.30 or R>0.30 and p<0.05), with greater correlation coefficients in LUAD compared with LUSC  at their expression levels. Within the cutoff values of R<-0.30 or R>0.30 and p<0.05, we identified LIMD1 mRNA abundance significantly correlates with IL6R and TNFSF15, and negatively correlates with TNFSF4 and TNFSF18. LIMD1 also negatively correlates with the MHC molecular TAP1  rates of cancer patients was analyzed in different portals, including TIMER2, TISIDB, GEPIA2, and DriverDB portals with TCGA dataset, and Kaplan Meier plotter (KMPlot) portal with TCGA, EGA, and GEO datasets. We found that the abundance of LIMD1 is significantly associated with overall survival (OS) of HPV-positive HNSC, KICH, KIRC, LGG, LUAD, and READ, and representative results from TIMER2 are shown in Regarding the prognostic implications of LIMD1 in NSCLC patients, similar results were obtained from TIMER2, TISIDB, DriverDBv3, and KMPlot portals showing that the LIMD1 expression levels positively correlate with the overall survival (OS) of LUAD but not that of LUSC patients   , VHL 4,,4, 5, TRFinally, we analyzed the possible pathways with the involvement of LIMD1 in KEGG and Uniport Figure 7In this algorithm-based study, we show that: 1) LIMD1 is significantly deregulated in several malignancies, including its downregulation in NSCLC Figure 2Overexpression of LIMD1 was shown to be a hallmark of ABC type of DLBC . Our GEOcis   –62. In lis-NATs) . Apart fLIMD1 has both cytoplasmic and nuclear roles, due to its ability to shuttle between the cytoplasmic and nuclear compartments. It is known to be engaged in the regulation of HIF1α, Wnt and Hippo, and cytoskeletal pathways in the cytoplasmic compartment , 14, 50,LIMD1 is heavily phosphorylated (PhosphoSitePlus.org), such as Y21, Y179, S233, S272, S277, S316, S421, S424, Y527 in humans, with a few sites experimentally validated that render its different functions . Cell-cyBesides these mechanisms, disclosing other potential unknown mechanisms underneath context-dependent LIMD1 deregulation and its promo-oncogenic and anti-oncogenic roles in cancers will be our future priority of study. Importantly, our recently published findings have shown that LIMD1 is associated with DNA damage response and protects B lymphoma cells from DNA damage-induced cell death, which represents a novel mechanism accounting for its oncogenic properties in this setting , 64. Of In summary, this meta-analysis indicates that LIMD1, as a tumor suppressor in lung cancer, is significantly downregulated in NSCLC, and could serve as a prognosis marker for LUAD but not for LUSC. This study represents a breath of cancer patient sample databases, and discloses various potential mechanisms underneath the interaction of LIMD1 with NSCLC, for example, site-specific phosphorylation at S233, S272, and S277 of LIMD1 and LIMD1-mediated immune infiltration in this setting. These underlying molecular mechanisms of LIMD1-mediated anti-oncogenesis in NSCLC merit further experimental validation and elaboration, which will be our priority in the following pursuits.The original contributions presented in the study are included in the article/Conceptualization: LW, SN. Data analysis: SN. Funding acquisition: LW, SN. Methodology: LW, MH, SN. Writing and editing: LW, SN. All authors contributed to the article and approved the submitted version.This work was supported by NIH (CA252986 and DE027314) and ASH, and in part by the NIH grant C06RR0306551.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.


Renal toxicity is a serious side effect that hinders the use of cisplatin, a commonly used and effective chemotherapeutic agent. Meanwhile, quinacrine is an FDA approved drug that has been stated for its anti-inflammatory effect. Thus, we investigated the ameliorative effect of quinacrine against cisplatin-induced renal toxicity. Single intraperitoneal (i.p.) 10 mg/kg cisplatin administration induced renal injury in rats. Our results showed that 10 mg/kg/day quinacrine decreased the mortality rate of rats from 46.15% (cisplatin group) to 12.5%, and significantly decreased renal tissue fibrosis, relative kidney to body weight ratio, serum creatinine and urea levels compared with the cisplatin group. Indeed, quinacrine significantly decreased renal malondialdehyde concentration and increased renal total antioxidant capacity, compared with the cisplatin group. Furthermore, quinacrine caused significant upregulation of renal sirtuin-1 (SIRT-1) with significant downregulation of intercellular adhesion molecule-1 (ICAM-1) and tumor necrosis factor-α (TNF-α). Moreover, quinacrine significantly blocked cisplatin-induced apoptosis, which was made evident by downregulating renal apoptotic proteins (BAX and p53) and upregulating the renal anti-apoptotic protein BCL2, compared with the cisplatin group. In conclusion, this study demonstrates, for the first time, that quinacrine alleviates cisplatin-induced renal toxicity via upregulating SIRT-1, downregulating inflammatory markers (ICAM-1 and TNF-α), reducing oxidative stress, and inhibiting apoptosis. Cisplatin is an efficient chemotherapeutic agent in in the treatment of different types of cancers, including non-small cell lung carcinoma, cervical, ovarian, testicular and head and neck cancers ,2,3. Unf+ dependent histone deacetylase, has recently gained considerable attention, due to its differential regulation of inflammation, stress response and apoptosis [It has been shown that cisplatin induces renal injury via increasing oxidative stress levels, mitochondrial dysfunction, inflammation, DNA damage and apoptosis . Sirtuinpoptosis ,9. SIRT1poptosis ,11. Thuspoptosis .Quinacrine is an FDA-approved antimalarial drug and a well-known anti-inflammatory agent. It has also been reported to be beneficial in managing cancer, lupus erythematosus, and cutaneous sarcoidosis ,14,15. MCisplatin induced mortality in 46.2% of rats A along wCisplatin induced oxidative stress in renal tissues, as it significantly increased renal malondialdehyde (MDA) concentration E and sigCisplatin induced tubular dilation lined with flattened epithelium and vacuolar degeneration in the renal cortex A, and maCisplatin significantly decreased renal SIRT-1 concentration A and sigCisplatin caused apoptosis in renal tissue, which was indicated by the significant increase in immunostaining of apoptotic proteins BAX and p53 A,B, compNephrotoxicity is a serious side effect of cisplatin that limits its clinical use. This study aims to investigate the potential protective effect of quinacrine, an FDA-approved drug, against cisplatin-induced nephrotoxicity, in addition to studying the possible underlying molecular mechanism of quinacrine’s action.Single injection of cisplatin in rats induced nephrotoxicity, which was evident by the increase in mortality rate, relative kidney to body weight ratio, serum creatinine and urea concentrations, fibrosis formation and structural alterations of renal cortex and medulla. This was in agreement with El-Sherbiny et al. . MoreoveQuinacrine was also previously reported to have an antioxidant effect in ulcerative colitis , status As previously shown, inflammation plays a major role in the pathogenesis of cisplatin-induced renal injury ,29. ConsRecent studies revealed that SIRT-1 upregulation alleviates apoptosis in drug-induced injuries ,35,36. AIn this study, cisplatin led to a significant increase in expression levels of caspases, BAX and p53 with a significant decrease in expression levels of BCL2. These results suggest that SIRT-1 downregulation might play a regulatory role in cisplatin-induced nephrotoxicity. Consistently, Kim et al. (2019) stated that SIRT-1 activation attenuates cisplatin-induced cell apoptosis, probably through deacetylating p53 . QuinacrAltogether, cisplatin downregulated SIRT-1 levels, which might be—in part—due to induction of oxidative stress. SIRT-1 downregulation resulted in an increase in pro-inflammatory markers (ICAM-1 and TNF-α) in addition to upregulation of p53 and pro-apoptosis proteins (BAX and caspases). It also led to downregulation of the anti-apoptosis protein BCL2. Quinacrine reversed these effects, which efficiently protected against cisplatin-induced nephrotoxicity C.Twenty-nine male Sprague Dawley rats weighing (180–200 gm) were purchased from the Faculty of Pharmacy, Mansoura University, Egypt. Rats were allowed to be acclimated by keeping them under standard conditions of a 12 h dark and 12 hr light cycle, 23–27 °C temperature, and 48–52% humidity for a week before administration of the first dose. Rats were provided with the standard pellet diet and water ad libitum. The animal care and experiments described in this study complied with “Research Ethics Committee” Faculty of Medicine, Mansoura University, Mansoura, Egypt, which is in accordance with “Principles of Laboratory Animal Care”  in animal care and experiments.Cisplatin (232120) and quinacrine (Q3251) were purchased from Sigma-Aldrich . All other chemicals used in the study were of high purity and analytical grade.Cisplatin was dissolved in saline. Acute renal toxicity was induced by intraperitoneal injection of 10 mg/kg cisplatin, a single dose, at day 5 ,44.Normal group (8 rats): rats were injected with 0.2 mL saline intraperitoneally, daily for 10 days.Cisplatin group (13 rats): rats were intraperitoneally injected with 0.2 mL saline daily for 10 days, except at day 5, when they were injected with cisplatin single-dose intraperitoneally (10 mg/kg).Cisplatin + quinacrine (8 rats): rats were intraperitoneally injected with quinacrine (10 mg/kg/day) for 10 days ,21,42,45Rats were randomly divided into three groups:The mortality of the rats was daily recorded . At the Rats were sacrificed to dissect the kidneys. The kidneys were rinsed with cold phosphate buffer saline (PBS) pH:7.4, blotted dry with filter paper, and then weighed. A transverse cut of the right kidney was fixed in 10% phosphate-buffered formalin (PBF) for histopathological and immunohistochemical analyses. The left kidney was homogenized in cold PBS, centrifuged, and then the supernatant was aliquoted and kept at −80°C for further oxidative stress and ELISA analyses.Serum was used for the determination of creatinine (#10053) and urea (#10505) concentrations, according to Bartles et al.,  and FawcA supernatant of the kidney tissue homogenates was used for determination of the MDA (# MD 25 29) concentration, following the method of Kei et al.  and TAC A supernatant of the kidney tissue homogenates was used for determination of SIRT-1, ICAM-1, and TNF-α concentrations, according to the manufacturer’s instructions, using the following ELISA kits: CSB-EL021339RA, CSB-E04576r and CSB-E11987r, respectively . Briefly, standards and samples were added to a 96-well plate, covered with adhesive strip, and incubated for 2 h at 37 degrees. Liquids were removed from all wells without washing. Biotin-antibody (1×) was added to each well, covered with a new adhesive strip and incubated for 1 h at 37 °C. The antibody was removed and the wells were washed three times, using the provided washing buffer. HRP-avidin (1×) was added to each well, and the microtiter plate was covered with a new adhesive strip and incubated for 1 h at 37 °C. The solution was aspirated, and the wells were washed five times. TMB substrate was added to each well and incubated for 20 min at 37 °C in dark. STOP Solution was added to each well, and the optical density was measured, using a microplate reader at 450 nm.The formalin-fixed kidney tissues were embedded in paraffin. Sections were cut into 5 μm in thickness and stained with hematoxylin and eosin (H&E). Slides were used for renal morphological and structural alteration examination under light microscopy. Pictures were captured, using a digital camera. The slides were blindly examined.For % area of fibrosis, the sections were stained with Masson’s trichrome for examining the fibrosis formation and assessment of % area of fibrosis, as Masson’s trichrome stains fibrotic areas with blue and parenchymal cells with red.-3 , Caspase-8 , Caspase- 9 , BAX , BCL2  and p53  at a 1:100 dilution factor. The slides were rinsed three times with PBS, and incubated with appropriate secondary antibodies for 30 min at 25 °C. As a negative control, the primary antibody was replaced by normal rat serum. The specificity of the used antibodies was checked using No antibody control, secondary anti-mouse and secondary anti-rabbit controls are showing negative staining to confirm that the reported signal was antigen specific . The sections were treated with 3 % hydrogen peroxide and protein block, followed by overnight incubation at 4 °C with the following antibodies: TNF-α , Caspase-1 , Caspasespecific . For the® digital camera installed on an Olympus® microscope with 1/2 X photo adaptor, using 400× objective. The images were then analyzed on an Intel® Core I3® based computer, using VideoTest Morphology® software  with a specific built-in routine for area, % area measurement and object counting. Masson’s trichrome blue color stained areas, Caspase-1, -3, -8, and -9, and BCL2 and p53 brown color immunostained areas were blindly determined, using ImageJ software , taking six reads for each section at 400× magnification. Briefly, digital image analysis was performed by uniformly adjusting size of all images to 12.7 centimeters in width and 9 centimeters in length (300 dpi). In this case, a scale bar was set to (25 microns). All images were loaded into the (image J) program, where the 6 fields were investigated. Finally, the average area and % area for each group were calculated relative to the control normal group.The slides for Masson’s trichrome and immune-histochemical stainings were photographed, using an Olympusp ≤ 0.05 was considered statistically significant, with the following used symbols: * p < 0.05; ** p < 0.01; *** p < 0.001.All data are presented as mean ± standard error of the mean (S.E.M). Statistical analysis was performed via GraphPad Prism V 5.02 . The distribution of data was analyzed to check the parametric or non-parametric tests, which will follow. One-way ANOVA followed by Bartlett’s test (post-test) was used to determine the statistical significance between groups. The current study proposes, for the first time, that quinacrine ameliorated cisplatin-induced renal toxicity via upregulation of SIRT-1. Moreover, no previous studies, to our best knowledge, have reported the effect of quinacrine on SIRT-1 regulation. Furthermore, quinacrine significantly downregulated the pro-inflammatory proteins (ICAM-1 and TNF-α), restored the cisplatin-disrupted oxidative balance, suppressed the apoptotic mediators , and upregulated the anti-apoptotic protein BCL2. Although clinical studies are needed to establish the renoprotective effect of quinacrine against cisplatin-induced renal toxicity, this study provides evidence for a potential novel therapeutic use of quinacrine as a protective agent against cisplatin-induced renal injury.


The signs and symptoms of MGD were evaluated by ocular surface disease index (OSDI), tear film breakup time (TBUT), and meibomian gland yield secretion score (MGYSS). The clinical evaluators and tear samples were analyzed at baseline and at each IPL treatment session. Concentrations of (chemokine ligand) CXCL1, (C-C motif chemokine) CCL11, (tumor necrosis factor) TNF-α, (interferon) IFN-γ, (interleukin) IL-2, IL-6 and  TIMP-1were measured by Quantibody Human Dry Eye Disease Array1. OSDI significantly decreased after IPL treatment compared with baseline. TBUT and MGYSS increased consecutively during treatment. CXCL1, CCL11, TNF-α, IFN-γ, IL-2, IL-6 presented significantly decrease and TIMP-1 showed significantly increase from the pretreatment baseline. The changed concentrations of TNF-α, IFN-γ, IL-2, TIMP-1 correlated with TBUT, the changed values of CXCL1, TNF-α, IFN-γ, CCL11, IL-2, IL-6, TIMP-1 correlated with MGYSS, and the changed concentrations of CXCL1, IFN-γ, CCL11, IL-2, IL-6 correlated with TIMP-1. The data supported IPL treatment could significantly relieve both signs and symptoms of MGD. The therapeutic effect of IPL treatment may originate from regulation of inflammatory cytokines including CXCL1, TNF-α, IFN-γ, CCL11, IL-2, IL-6, and TIMP-1.Meibomian gland dysfunction (MGD) has become a prevalent ocular surface disorder. Its pathogenesis is regarded as a self-perpetuating inflammatory vicious circle. Intense Pulsed Light (IPL) treatment was recently applied to improve the meibomian gland function and reduce symptoms of MGD. However, studies investigating the change of specific inflammatory cytokines during IPL treatment remained sparse. To further figure out how IPL treatment modulates the inflammatory cytokines in tears of MGD, we therefore performed a cross-sectional study and enrolled 32 patients from March 2019 to December 2020. The patients received 3 sessions of IPL treatment (10 to 16 J/cm Dry eye disease (DED) represents a multifactorial disease of ocular surface characterized by a loss of homeostasis of tear film . BecauseTreatments for MGD include topical antibiotic and anti-inflammatory agents, tear substitutes, warm compress and meibomian glands massage . HoweverDemodex [Recently, intense pulsed light (IPL) has been applied as a novel therapeutic measure for MGD. IPL treatment could ameliorate symptoms and improve the condition of the tear film with relatively long-term effectiveness –11. DespDemodex , decreasDemodex , 18 playHowever, studies investigating the change of specific inflammatory cytokines during IPL treatment remain sparse. To further figure out how IPL treatment interferes the inflammatory vicious circle of MGD, we analyzed tear cytokines including (chemokine ligand) CXCL1, (C-C motif chemokine) CCL11, (tumor necrosis factor) TNF-α, (interferon) IFN-γ, (interleukin) IL-2, IL-6 and  TIMP-1. In addition, the clinical correlations with these cytokine levels were also analyzed.Patients were enrolled from the outpatient department of The Hospital of Ningxia Hui Autonomous Region from March 2019 to December 2020. The cross-sectional study adhered to the tenets of the Declaration of Helsinki and was approved by the ethics committee of the hospital. Written informed consent was obtained from all patients. Authors had access to information that could identify individual participants during data collection. Patients over 18 years of age and met diagnostic criteria of MGD , (8) wit2 depending on the Fitzpatric skin type [Patients received 3 sessions of IPL treatment with the M22 system  at 4-week interval. The treatment was performed by a same doctor. The pulse intensity raged from 10 to 16 J/cmkin type , pulse wThe clinical evaluations were performed at baseline and at each IPL treatment session. All measurements were conducted prior to IPL treatment. Subjective symptoms were estimated by the ocular surface disease index (OSDI). Tear film break-up time (TBUT) was facilitated by observing with a blue filter after applying fluorescein-strip into the inferior conjunctival sac. TBUT was measured 3 times for each patient and average was calculated. Meibomian gland yield secretion score (MGYSS) was used to scored meibum quality  . The scoTear samples were collected before any other operations. The samples were collected after instillation of 60μL of phosphate-buffered saline into the inferior conjunctival sac. The patients moved eyeballs to mix the tear fluid content before sampling . The samP < 0.05 was statistically significant.Data were expressed as mean ± standard error of the mean (SEM). The data of tear cytokines were also transformed into ratios by using the baseline value as the reference. Shapiro–Wilk test with SPSS 26.0 for Windows Software  was used to test the normality of data. A Paired Samples T Test was used to compare the baseline and posttreatment cytokine concentration after each IPL treatment session. Correlations between the tear cytokines and clinical parameters, and between their changed values after IPL treatment were analyzed by Spearman correlation coefficient, respectively. 32 eyes of 32 patients  were included into the study, the visual acuity and intraocular pressure of patients kept stable during IPL treatment. The mean (±SD) age of all the patients was 54.13±8.74 years . OSDI decreased after IPL treatment compared with baseline, which were of statistically difference. TBUT and MGYSS increased consecutively during treatment. The clinical evaluators were listed in P = 0.07). The concentration of tear cytokines mentioned above was showed in In the study we used a highly sensitive antibody microarray approach for the quantification of cytokines in tears of patients with MGD. The concentrations of CXCL1, TNF-α, IFN-γ, CCL11, IL-2, IL-6 presented significantly decrease and TIMP-1 showed significantly increase from the pretreatment baseline. CCL11 slightly raised after the second IPL treatment, however, the difference was not statistically significant . The changed concentrations of CXCL1, TNF-α, IFN-γ, CCL11, IL-2, IL-6 after the entire IPL treatment correlated well with the increase of MGYSS . The correlations between the changed concentrations of CXCL1, IFN-γ, CCL11, IL-2, IL-6 after the entire IPL treatment and decrease of OSDI showed statistically significance . After the whole IPL treatment, the correlation analysis between the changed values of TIMP-1 and the changed values of TBUT, MGYSS and OSDI also showed statistically significance .A correlation between the improvement of TBUT and decrease of TNF-α, IFN-γ, IL-2 after the entire IPL treatment was observed  activation and limit fibrosis , 44. SevIn our study, IL-6, IFN-γ and TNF-α decreased significantly following IPL treatment in MGD patients, and the decrease of these cytokines was similarly correlated strongly with the improvement of TBUT, MGYSS and OSDI. These findings consistent with that of previous studies, demonstrated that IPL treatment enabled to significantly downregulated inflammation which is the core mechanism of MGD. The possible reason may be that IPL selectively ablates the superficial vessels located in eyelid margin, prevents the continued secreting of cytokines. In addition, the thermal interaction of IPL may also make the meibum less viscous, thus facilitate meibomian gland expression and unclogging of the gland. And last, the natural tendency of the eyelids to lose rigidity and elasticity may be reversed by the photomodulation of IPL, which assisted in returning eyelids to normal position and accomplishing complete blinks . The proInterestingly, we observed significant elevation of TIMP-1 in the current study. More importantly, the improvement of TBUT, MGYSS and OSDI was significantly correlated with the increase of TIMP-1. TIMP-1 is a natural collagenase inhibitor of MMP9, and has recently emerged as a decisive factor in several human pathologies . MMP-9 oThere are several limitations to our study. First, the study design was observational case series and lack of a control group. Second, the sample size was small and the duration of follow-up was limited to 4 weeks after final IPL treatment. Third, the patients received IPL treatment immediately followed by meibomian glands expression, which may help to clear clogged meibomian glands duct. However, previous studies have confirmed the improvement of dry eye symptoms and signs after IPL treatment alone. Furthermore, randomized controlled studies with a larger number of patients and longer follow-up period are necessary to assess the long-term effectiveness and safety of IPL treatment. Meanwhile, more exploratory studies are required to elucidate the function and potential mechanism of different inflammatory cytokines in tears of MGD.In conclusion, the study supported the efficacy of IPL treatment in relieving both signs and symptoms of MGD. The therapeutic effect of IPL treatment may originate from regulation of inflammatory cytokines including CXCL1, TNF-α, IFN-γ, CCL11, IL-2, IL-6 and TIMP-1.


Arabidopsis. Specifically, NAC102 localizes in chloroplast nucleoids. Yeast two-hybrid assay and co-immunoprecipitation assay suggested that NAC102 interacts with chloroplast RNA polymerases. Furthermore, overexpression of NAC102 in chloroplasts leads to reduced chloroplast gene expression and chlorophyll content, indicating that NAC102 functions as a repressor in chloroplasts. Our study not only revealed that transcription factors are new regulators of chloroplast gene expression, but also discovered that transcription factors can function in chloroplasts in addition to the canonical organelle nucleus.The chloroplast is a semi-autonomous organelle with its own genome. The expression of chloroplast genes depends on both chloroplasts and the nucleus. Although many nucleus-encoded proteins have been shown to localize in chloroplasts and are essential for chloroplast gene expression, it is not clear whether transcription factors can regulate gene expression in chloroplasts. Here we report that the transcription factor NAC102 localizes in both chloroplasts and nucleus in  One of the major differences between plants and animals is that plants contain chloroplasts, which carry out photosynthesis, produce various metabolites, and sense external cues. Chloroplasts are believed to derive from cyanobacterium in an endosymbiotic event. During evolution, most bacteria genes were transferred to the host nucleus and only a small portion of genes were retained on the own genome ,2. TherepsbA, psbB, and rbcL, is preferentially dependent on PEP, the expression of the housekeeping genes, including ribosomal RNAs and the core subunits of the PEP, is dependent on NEP [rpoTp and rpoTmp, in Arabidopsis. PEP is a bacteria-type RNA polymerase that consists of four core subunits , encoded by rpoA, rpoB, rpoC1, and rpoC2 in the chloroplast genome, and a promoter-recognizing sigma factor (σ) encoded by the nuclear genes [As a semi-autonomous organelle, the gene expression in chloroplasts is controlled by both chloroplasts and the nucleus . For exat on NEP ,8,9. NEPar genes ,11. Furtar genes ,13,14.In addition to PAPs, many other nucleus-encoded proteins have been reported to regulate chloroplast gene expression at the transcriptional level or the post-transcriptional level  ,15,16. Anac102 mutant is sensitive to excessive light [In plants, the NAC  family is one of the largest plant-specific transcription factors and plays important roles in plant development and stress responses ,18. Prevve light ,20,21. HArabidopsis thaliana used in this study were in the Columbia (Col-0) background. The nac102 mutant (SALK_030702) was obtained from ABRC. The transgenic lines were generated by the floral dip method [Arabidopsis was grown on half-strength (1/2) Murashige and Skoog (MS) medium or in soil under long-day conditions (16 h of light and 8 h of dark) at 22 ± 2 °C.All p method . ArabidoΔN43-eYFP was generated by using NAC102-eYFP as a template. To generate NAC102N43-eYFP, NAC102N60-eYFP, and NAC102N80-eYFP, the 43, 60 and 80 aa N-terminal parts of NAC102 were cloned and fused to eYFP, respectively. To generate NAC102-eYFPNES, the coding sequence of nuclear export sequence (NES) (CLPPLERLTLD) [TRXz. All fusion fragments were inserted into pFGC5941 at the NcoI and XbaI sites. For Y2H assays, the coding sequence of NAC102 was inserted into pGBKT7 at the NdeI and PstI sites. The coding sequences of the subunits of PEP and NEP were inserted into pGADT7 at the NdeI and PstI sites. All recombination reactions were performed using a Lightening Cloning kit . The primers used for vector constructions are listed in the For localization analysis of NAC102, the coding sequence of enhanced yellow fluorescent protein (eYFP) was fused to the C-terminus of the NAC102 fragment (NAC102-eYFP) using fusion PCR. The NAC102LERLTLD)  was furtArabidopsis protoplasts, the plasmids were transfected into protoplasts as described previously [For transient expression in eviously  and incuImages were captured using a confocal laser scanning microscope . The excitation wavelengths for DAPI, CFP, eYFP, and chlorophyll autofluorescence were 405 nm, 448 nm, 488 nm, and 514 nm, respectively. The emission filters were substrate for DAPI, 448–514 nm for CFP and eYFP, and 500–530 nm for chlorophyll autofluorescence.The pGADT7 vectors were transformed into yeast strain AH109, while the pGBKT7 vectors were transformed into yeast strain Y187. After mating AH109 with Y187, the yeasts were plated on DDO media. The colonies were resuspended and diluted using distilled water, and were then plated on DDO or QDO media, respectively. The photos were taken after incubation at 28 °C for 2 d on DDO media or 3 d on QDO media.g for 5 min, the resulting supernatants were incubated with 40 µL GFP-Trap beads  for 1 h. After washing three times with washing buffer , the beads were incubated with 40 µL 2× SDS-PAGE buffer for 10 min at 98 °C. The eluted proteins were subjected to Western blotting using anti-GFP , anti-rpoA , anti-rpoB , anti-rpoC1 , anti-rpoC2 , anti-rpoTp , and anti-rpoTmp .The 8-day-old seedlings were ground in liquid nitrogen and incubated with IP buffer  for 30 min. After centrifugation at 12,000× Arabidopsis seedlings using TRIZOL reagent . The reverse-transcription reaction was performed using HiScript II Q RT SuperMix  according to the manufacturer’s instructions. The qPCR assays were performed on the CFX96 Touch Real-Time PCR Detection System  using ChamQ Universal SYBR qPCR Master Mix . The primers used for qPCR analyses are listed in Total RNA was extracted from 8-day-old Chlorophyll contents were measured as described previously . The 3rdArabidopsis thaliana, there are 105 NAC transcription factors [AT5G63790) is predicted to localize in both chloroplast and nucleus according to TAIR . To our knowledge, no transcription factors have been reported to localize in the chloroplast. Therefore, we decided to study NAC102 further. NAC102 has two alternative transcripts, AT5G63790.1 and AT5G63790.2. As shown in AT5G63790.1, AT5G63790.2 encodes a protein with an additional 10 amino acids at the N-terminus. To confirm the subcellular localization of NAC102, both NAC102 variants were fused with enhanced yellow fluorescent protein (eYFP) and were transiently expressed in Arabidopsis protoplasts. The CFP-tagged E2Fa transcription factor (E2Fa-CFP) was used as the nucleus marker and the chlorophyll fluorescence was used as the chloroplast marker. As shown in AT5G63790.1. To further confirm its localization, we generated the transgenic Arabidopsis expressing NAC102-eYFP (AT5G63790.1) driven by the 35S promoter (35S:NAC102-eYFP). Consistently, the eYFP fluorescence was detected in both chloroplast and nucleus  in Arabidopsis protoplasts. In support of the hypothesis, the NAC102ΔN43-eYFP was only detected in the nucleus  as a bait. The coding sequences of the subunits of PEP  and NEP (rpoTp and rpoTmp) were fused to the GAL4 activation domain (AD) as prey. Compared with the AD control, the expression of PEP and NEP subunits promoted yeast growth on the selection media (QDO + 3-AT), indicating that NAC102 interacts with PEP and NEP subunits , which localized in chloroplasts, but not in the nucleus , which triggered cell death. Although NAC102 functions as a repressor, other transcription factors in chloroplast may function as activators. Therefore, our study suggests that it is possible to regulate chloroplast gene expression using transcription factors, which may represent a new strategy to manipulate chloroplasts.Our data suggested that NAC102 represses chloroplast gene expression. When NAC102 was overexpressed in chloroplasts, both chloroplast gene expression and the chlorophyll contents were reduced . Howevernac102 . One post stress . Althoug


Atherosclerosis is considered the major cause of cardiovascular and cerebrovascular diseases, which are the leading causes of death worldwide. Excessive nitric oxide production and inflammation result in dysfunctional vascular endothelial cells, which are critically involved in the initiation and progression of atherosclerosis. The present study aimed to identify a bioactive compound from Jerusalem artichoke leaves with anti-inflammatory activity that might prevent atherosclerosis. We isolated bioactive heliangin that inhibited NO production in LPS-induced macrophage-like RAW 264.7 cells. Heliangin suppressed ICAM-1, VCAM-1, E-selectin, and MCP-1 expression, as well as NF-κB and IκBα phosphorylation, in vascular endothelial cells stimulated with TNF-α. These results suggested that heliangin suppresses inflammation by inhibiting excessive NO production in macrophages and the expression of the factors leading to the development of atherosclerosis via the NF-κB signaling pathway in vascular endothelial cells. Therefore, heliangin in Jerusalem artichoke leaves could function in the prevention of atherosclerosis that is associated with heart attacks and strokes. Cardiovascular diseases (CVDs) are the most prevalent cause of death, accounting for ~31% of all deaths worldwide ,3,4. ArtJerusalem artichoke is a sunflower native to North America, and its tubers are consumed worldwide . They coThe inflammation of VECs causes arteriosclerosis. The expression of intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion protein 1 (VCAM-1), and E-selectin increases in VECs during the initiation and progression of arteriosclerosis. Chemoattractant protein-1 (MCP-1) promotes the migration of monocytes and macrophages into VECs. The cell adhesion molecules L-, P-, and E-selectin are expressed in leukocytes, platelets, and endothelial cells, respectively, during the early phase of inflammation ,17,18,192 incubator + Δ−0.3 mmu. The NMR and mass spectrometry data further substantiated that fraction 6 was heliangin. Heliangin has been identified in Bejaranoa semistriata, Viguiera eriophora, and Viguiera puruana [Nitric oxide is a mediator and regulator of inflammatory responses . We prevand COSY . The cheand COSY  on 13C-  puruana ,32. We assessed the ability of purified 2, 5, 10, 25, and 50 µM heliangin extracted from Jerusalem artichoke leaves to inhibit NO production. The results showed that 50 µM heliangin was slightly cytotoxic to RAW 264.7 cells, whereas a low concentration was not b. Furthe2 levels in macrophages cells lysate via the MAPK and NF-κB pathway [The bioactivity and cytotoxicity of heliangin have been investigated in Ehrlich ascites tumor cells, as well as in human oral epidermal carcinoma, cervical epithelial cells, and liver carcinoma (hepa59T/VGH) ,33. Appr pathway . Our fin pathway  did not ICAM-1, VCAM-1, E-selectin, and MCP-1 are biomarkers of endothelial dysfunction in atherosclerosis ,41,42. WAtherosclerosis causes vascular disease, which is the leading cause of mortality worldwide. Therefore, we screened compounds from Jerusalem artichoke leaves for compounds with anti-inflammatory activity that might prevent atherosclerosis. We isolated a compound with potential anti-inflammatory activities using open column chromatography and purified it by C18 HPLC. We then confirmed that the compound was heliangin by NMR and mass spectrometry. We showed that heliangin inhibited NO production in RAW 264.7 cells induced by LPS and suppressed gene expression of ICAM-1, VCAM-1, E-selectin, and MCP-1 in VECs induced by TNF-α. Heliangin also suppressed NF-κB and IκBα phosphorylation induced by TNF-α in VECs. Therefore, heliangin appeared to suppress ICAM-1, VCAM-1, E-selectin, and MCP-1 induction by TNF-α in VECs via the NF-κB signaling pathway. These results suggest that heliangin derived from Jerusalem artichoke leaves could play a role in the prevention of inflammatory in macrophage and VECs, which might help to prevent atherosclerosis.


LMNA gene exon 2, variant c448A > C (p.Thr150pro), heterozygous form, AD).Emery–Dreifuss muscular dystrophy (EDMD) is a rare genetic disease that affects the musculoskeletal system, including the heart, causing rhythm disorders and cardiomyopathy, sometimes requiring an implantable cardioverter-defibrillator (ICD) or heart transplantation due to severe heart damage. The case described herein concerns a 16-year-old girl, with grade II obesity, without other known pathological antecedents or cardiac pathology diagnosis given an annual history of cardiological investigations. She was admitted to the Infectious Diseases Department with SARS-CoV-2 virus infection. The anamnesis showed that the cardiological investigations performed in the past were completed due to the medical history antecedents of her sister, who had been diagnosed with dilated cardiomyopathy, having undergone the placement of an ICD and a heart transplant. Numerous investigations were performed during hospitalization, which revealed high levels of high-sensitive cardiac troponin I (hs-cTnI), creatine kinase (CK) and N-terminal pro b-type natriuretic peptide (NT-proBNP). Dynamic electrocardiographic evaluations showed ventricular extrasystoles, without clinical manifestations. The patient presented stage 2 arterial hypertension (AHT) during hospitalization. A cardiac ultrasound was also performed, which revealed suspected mild subacute viral myocarditis with cardiomyopathy, and antihypertensive medication was initiated. A heart MRI was performed, and the patient was diagnosed with dilated cardiomyopathy, refuting the suspicion of viral subacute myocarditis. After discharge, as the patient developed gait disorders with an impossible heel strike upon walking and limitation of the extension of the arms and ankles, was hospitalized in the Neurology Department. Electrocardiograms (ECGs) were dynamically performed, and because the rhythm disorders persisted, the patient was transferred to the Cardiology Department. On Holter monitoring, non-sustained ventricular tachycardia (NSVT) was detected, so antiarrhythmic treatment was initiated, and placement of an ICD was subsequently decided and was diagnosed with EDMD. Genetic tests were also performed, and a mutation of the lamin A/C gene was detected ( SARS-CoV-2 virus infection may cause viral myocarditis, with the literature claiming that the SARS-CoV-2 virus is a trigger for myocarditis, but there are no clinical data to support the link between the SARS-CoV-2 virus and the unfavorable evolution of the Emery–Dreifuss syndrome (EDMD) ,2.EMD) and lamin A/C (LMNA) genes, and can be autosomal dominant or recessive [LMNA gene [EMD and negative LMNA, studies also incriminate the FHLA1 gene, which is associated with the EDMD6 phenotype, which is X-linked and includes hypertrophic cardiomyopathy [Emery–Dreifuss syndrome is a rare genetic disease, caused by mutations of emerin , which is an X-linked recessive disease [ disease .LMNA mutations are associated with autosomal dominant and autosomal recessive Emery–Dreifuss muscular dystrophy, Hutchinson–Gilford progeria syndrome limb girdle muscular dystrophy 1B, dilated cardiomyopathy with conduction defects, Charcot–Marie–Tooth axonal neuropathy type 2, familial partial lipodystrophy, mandibuloacral dysplasia, and atypical forms of Werner syndrome [syndrome ,5,6.LMNA mutations, as follows: 23 of them cause autosomal dominant Emery–Dreifuss muscular dystrophy (EDMD2), 8 of them cause dilated cardiomyopathy and 1 mutation causes autosomal recessive Emery Dreifuss (EDMD3) [There are 41 known  (EDMD3) ,8.LMNA-related dilated cardiomyopathy is characterized by left ventricular enlargement, reduced systolic function and arrhythmias, having unfavorable prognosis, with risk of sudden death [en death .LMNA, this being a rare condition, with estimated incidence of 3:1,000,000 [EDMD is characterized by a clinical triad including muscle–joint illness, muscle weakness and cardiomyopathy with rhythm disorders due to a structural or functional defect of the genes encoding the proteins of the nuclear envelope, e.g., ,000,000 ,10.This clinical case is of a 16-year-old female patient with unknown personal pathological history, with a cardiac hereditary history. Her sister was diagnosed with dilated cardiomyopathy, requiring ICD and heart transplant. The patient was hospitalized for SARS-CoV-2 virus infection, with mild COVID-19 symptoms including rhinorrhea and a dry cough.2 99% at room temperature, rhythmic heart sounds, without retrosternal pain, tachycardia, or tachypnea. The patient presented stage 2 hypertension , a heart rate of 80 beats per minute, normal ECG, without signs of meningeal irritation, and the Glasgow Coma Scale score was 15/15.At hospitalization, the clinical examination revealed a good general condition, with a temperature of 37.4 °C, SaO-Elevated values: high-sensitive cardiac troponin I ; creatine kinase ; N-terminal probrain natriuretic peptide ; D-dimers ; C-reactive protein ; aspartate aminotransferase ; alanine transaminase , lactate dehydrogenase ;-9 cells/L), lymphocytes (2.37 × 109 cells/L), neutrophils 6.2 × 109 cells/L, platelets 360 × 109 cells/L, creatinine 0.56 mg/dL, sodium 135 mmol/L, potassium 4.2 mmol/L.Normal values: leukocytes  was 55–60%, the left ventricle (LV) dimensions were 53/35 mm, with hypokinesia in the lower wall, a suspicion of viral subacute myocarditis and cardiomyopathy. Thus, conversion enzyme inhibitor treatment and heart MRI were performed.During hospitalization, an antiviral treatment with lopinavir/ritonavir (400/100 mg q12h × 3 days) was initiated, and the patient was also prescribed anti-inflammatory medication (paracetamol 500 mg po bd × 7 days). An anticoagulant with prophylactic enoxaparin sodium (enoxaparin sodium 0.4 mL qd × 8 days), an antihypertensive with ACE inhibitors (Ramipril 2.5 mg po qd × 3 days) and a hepatoprotective drug (Silymarin 150 mg po td × 8 days) were also introduced. The antiviral treatment was stopped at the indication of the cardiologist after 3 days of administration.The heart MRI performed immediately after discharge refutes the suspicion of viral subacute myocarditis and confirms the diagnosis of dilated cardiomyopathy .Early gadolinium enhancement had a normal appearance, without evidence of intracavitary thrombosis or microvascular obstruction.The late gadolinium enhancement revealed a small contrast outlet with mid-myocardial topography at the level of the basal segments of the inter-ventricular septum, respectively, and a small focal contrast outlet at the lower point of insertion in the right ventricle on the left ventricle.T2 mapping: T2 value 49–50 ms .T1 mapping: T1 values were slightly increased, without highlighting the expansion of the extracellular compartment. .2—indexed value was probably underestimated due to the patient’s weight), with systolic function around the lower limit of the normal value (LVEF 58%).The LV was slightly dilated , in exon 2 of the LMNA gene was identified, confirming the diagnosis of Emery-Dreifuss Muscular Dystrophy.One month after discharge, the patient presented musculoskeletal system disorders, with gait disorders , Achilles’ tendon retractions and limitation of the extension of the arms and ankles but with present symmetrical deep tendon reflexes. Therefore, she was admitted to the Neurology Department and genetic tests were performed. The heterozygous mutation c.448A > C  block, non-sustained ventricular tachycardia (NSVT) and isolated and systematized monomorphic extra-systolic ventricular arrhythmia in episodes of ventricular bigeminy were present throughout the recording .Cardiac ultrasound revealed situs solitus, levocardia, normal atrio-ventricular connections, and normal pulmonary venous return (PVR), intact interatrial septum (IAS) and interventricular septum (IVS). A mild LV contractile dysfunction was observed, with LVEF of 53% and 0-I scale mitral regurgitation.The heart ultrasound values were the following: interventricular septum (IVS) = 8 mm, left ventricle posterior wall thickness (LVPWs) = 9 mm, right ventricular end-diastolic volume (RVEDV) = 54 mm, left ventricular end-diastolic volume (LVEDV) = 39 mm, left atrium (LA) = 34 mm, right ventricle (RV) = 25 mm, right atrium (RA) = 36 mm, and left ventricular ejection fraction (LVEF) = 53%.Thus, antiarrhythmic treatment with amiodarone was initiated during the treatment; the PR interval was prolonged, so it was decided to transfer the patient to the Institute of Cardiovascular Diseases and Transplantation in Târgu Mureş, a center specializing in ICD placement in children. In the absence of specific guidelines, it was decided by the multidisciplinary team that the benefits outweighed the risks, and the patient was scheduled for the implantation of a bicameral cardiac defibrillator.After ICD placement, ECG was performed.Studies show that chronic diseases, including muscular dystrophies, are risk factors for the development of severe cases of SARS-CoV2 virus infection . The patEMD expresses a variant of EDMD with a severe phenotype, and the missense variant expresses a normal or low level of emerin and supports a milder phenotype. The pathogenic variant LMNA gene, of the homozygous or compound heterozygous pathogenic form, causes a more severe form of the disease, as in this clinical case [The literature has not shown that there is an increased risk of rapidly unfavorable evolution of Emery–Dreifuss syndrome. Studies show that the severity of the disease depends on the genes expressed—the absence of emerin in patients with genetic defects of cal case .EMD and FHL1 pathogenic variants for asymptomatic patients because heterozygous women are at risk of developing heart disorders and muscular dystrophy syndromes [LMNA pathogenic variant. The studies have revealed that each child has a 50% chance of inheriting the pathogenic variant from one of their parents with AD-EDMD [Molecular detection can be determined in yndromes . Emery–D AD-EDMD ,9,10.It cannot be detected by the genetic variants if the disease is AR or AD . In thisEMD or FHL1; a heterozygous pathogenic variant in LMNA; or biallelic pathogenic variants in LMNA identified by molecular genetic testing [The clinical symptoms for a diagnosis of EDMD are: limb muscle wasting and/or weakness and elbow or neck/spine joint contractures and a hemizygous pathogenic variant in  testing .Molecular genetic tests may include a combination of gene-targeted testing (multgene panel or single gene testing) and comprehensive genomic testing , depending on the phenotype.As the phenotype of EDMD is broad, individuals with the distinctive features described in the Case Presentation above are likely to be diagnosed using gene-targeted testing, whereas those with atypical features in whom the diagnosis of EDMD has not been considered are more likely to be diagnosed using genomic testing ,12,13.The evolution of the disease was rapidly progressive, with an unfavorable prognosis: the patient, evaluated cardiologically annually, did not present musculoskeletal or cardiological disorders prior to SARS-CoV2 virus infection. However, a close correlation between COVID-19 infection and the onset or the manifestation of the latent disease cannot be excluded or confirmed.In this case, multidisciplinarity was very important, with the patient being diagnosed with Emery–Dreifuss syndrome about 3 months after SARS-CoV2 virus infection, and the severe evolution of the disease required ICD implants at about 4 months after SARS-CoV2 virus infection.


Furthermore, the expression levels of the TIP60, P300, PRDM9, KMT5B, and KMT5C genes encoding PTM regulators were up-regulated in 17M compared to 3M female mice (p < 0.05). These findings indicate that altered related regulatory enzymes and PTMs are associated with physiological ovarian aging in mice, which is expected to provide useful insights for the delay of ovarian aging and the diagnosis and treatment of female infertility.Post-translational modifications (PTMs) have been confirmed to be involved in multiple female reproductive events, but their role in physiological ovarian aging is far from elucidated. In this study, mice aged 3, 12 or 17 months  were selected as physiological ovarian aging models. The expression of female reproductive function-related genes, the global profiles of PTMs, and the level of histone modifications and related regulatory enzymes were examined during physiological ovarian aging in the mice by quantitative real-time PCR and western blot, respectively. The results showed that the global protein expression of Kbhb (lysineβ-hydroxybutyryllysine), Khib (lysine 2-hydroxyisobutyryllysine), Kglu (lysineglutaryllysine), Kmal , Ksucc (lysinesuccinyllysine), Kcr (lysinecrotonyllysine), Kbu (lysinebutyryllysine), Kpr (lysinepropionyllysine), SUMO1 (SUMO1 modification), ub (ubiquitination), P-Typ (phosphorylation), and 3-nitro-Tyr (nitro-tyrosine) increased significantly as mice aged. Moreover, the modification level of Kme2 (lysinedi-methyllysine) and Kac (lysineacetyllysine) was the highest in the 3M mice and the lowest in 12M mice. In addition, only trimethylation of histone lysine was up-regulated progressively and significantly with increasing age ( Ovarian aging is a complex biological process influenced by the interaction and gradual accumulation of internal and external factors ,2, whichDrosophila was significantly shortened when the H3K4 demethyase-retinol-binding protein 2 (RBP 2) was deficient [Post-translational modification (PTM) is an important functional regulation pattern in vivo, including acetylation, methylation, ubiquitination, glycosylation, lipidylation and phosphorylation . Currenteficient . TherefoIn this study, the level of histone modifications and the global profiles of PTMs  were examined during physiological ovarian aging in mice. Furthermore, we screened for differentially expressed PTM-related regulatory enzymes to evaluate whether gene expression and PTMs are associated with physiological ovarian aging.p < 0.05, p < 0.01, and p < 0.001). Moreover, the corpus luteum level was highest in 17M mice (p < 0.001). The mature follicles showed the most significant difference in 12M and 17M mice (p < 0.001). The coefficient of ovary weight decreased between 3M and17M .To confirm follicle depletion in the 3M, 12M, and 17M mice, we performed histological analysis of the ovaries. As seen in n = 10). The results showed that both of the relative mRNA expressions of AMH and MVH decreased significantly as mice aged . On the contrary, the aging marker genes p16 and p53 showed a significant increase among 12M and 17M mice compared with the 3M mice . These data suggest that the ovarian function of 3M, 12M, and 17M mice gradually decreases with age, which may reflect the different functional states of the ovary in the physiological aging process (ovarian function exuberant stage-ovarian function decline stage-ovarian function failure stage).We used RT-PCR assays to detect AMH and MVH expression in the ovaries of 3M, 12M, and 17M mice (n = 10). The results showed that the Kbhb (lysineβ-hydroxybutyryllysine), Khib (lysine 2-hydroxyisobutyryllysine), Kglu (lysineglutaryllysine), Kmal , Ksucc (lysinesuccinyllysine), Kcr (lysinecrotonyllysine), Kbu (lysinebutyryllysine), Kpr (lysinepropionyllysine), SUMO1 (SUMO1 modification), ub (ubiquitination), P-Typ (phosphorylation), and 3-nitro-Tyr (nitro-tyrosine) increased significantly as mice aged, respectively. Meanwhile, the Kme2 (lysinedi-methyllysine) and Kac (lysineacetyllysine) were highest in the 3M mice and lowest in 12M mice. However, there was no effect on the Kme3 (lysinetri--methyllysine), Klac (lysinelactyllysine), Kbz (lysinebenzoyllysine), or O-GlcNA (O-GlcNAcylation) .We used 18 pan-antibodies to test the changes of PTMs in the global protein during the ovarian aging of mice (n = 10). In this study, the levels of 15 PTMs, including Kme2, Kme3, Kac, Klac, Kbhb, Khib, Kglu, Kmal, Ksucc, Kcr, Kbz, Kbu, Kpr, ubiquitin, and O-GlcNA, were evaluated by western blot using a pan-antibody during ovarian aging (p < 0.001), H4 ubiquitination was obviously higher in the 12M and 17M mice than 3M (p < 0.001), whereas the Kpr and O-GlcNA level in 17M was significantly decreased compared with the level in 3M mice (p < 0.01).To identify whether PTMs of histones are involved in regulating gene expression in ovarian aging, we extracted the histones from the ovaries of 3M, 12M, and 17M mice . Furthermore, the results show that the increase in PTM level was positively correlated with the up-regulation of PTMs, regulating enzymes during ovarian aging in mice.The main functions of histones and global profiles of PTMs are regulated by serial transferases and demethylases. Given the above results, we next assessed whether this is related to the changes in expression of their regulatory enzymes. As shown in Ovarian aging is mainly affected by complex phased gene expression and transcription factor regulatory networks ,7,8,9,10In the present study, we measured PTMs in the ovaries of 3M, 12M, and 17M mice during the physiological aging process. The results showed that the modification level of PTMs and the expression levels of regulatory enzyme genes showed clear changes in the ovarian hypofunction group (12M) and the ovarian failure group (17M) compared with the adolescent group (3M), suggesting that changes in PTMs are associated with physiological senescence of the ovary. Most importantly, we first used 18 types of PTM pan-antibodies to detect changed PTMs during the global protein level in the physiological ovarian aging process. The results showed that the modification levels of three classical PTMs , nine novel PTMs , and the expression of regulatory enzymes, including p300 and TIP60, clearly increased. The level of glycolysis decreases in the p300 gene knockout group in HCT116 cells, which reveals that loss of p300 causes a decrease in 2-hydroxyisobutyrylation in glycolytic enzymes, thereby inhibiting glycolysis . HoweverHistones are the protein components of the nucleosome, which form the basic architecture of eukaryotic chromatin. The N terminus of histones H2A, H2B, H3, and H4 can undergo many types of post-translational modification, including methylation, phosphorylation, acetylation, and ubiquitination . HistoneMeanwhile, our sequencing analysis showed that the level of histone glycosylation and propionylation only decreased significantly in the 17M group, and no obvious change was observed in the 12M group. A previous study demonstrated that histone glycosylation is strongly associated with the metabolism of amino acids, fatty acids, and energy , while lIn conclusion, this study demonstrates that female reproductive function-related genes and the modification of Kme3, ubiqutin, Kpr, and O-GlcNA, as well as global levels of PTMs, including metabolism-related modification, phosphorylation, and nitro-tyrosine, are altered during physiological aging in the mouse ovaries, indicating the diverse and complicated regulation of gene expression and PTMs in age-induced female reproductive dysfunction. In addition, we found that KMT5C also shows a significant increase in the ovaries of mice treated with cyclophosphamide (120 mg/kg) and busulfan (12 mg/kg) at 2M, which were selected as pathological ovarian aging models. Together, these results suggested that KMT5C and Kme3 as-mediated epigenetic pathways may play a key role in the ovarian aging process, and their specific role is worthy of further study and exploration.Three-month-old, twelve-month-old, and seventeen-month-old KM female mice were housed at the Center of Experimental Animals, Nanchang University. The weights of the mice and ovaries were measured and recorded on the last day of treatment. All mice were housed in 12-h dark and 12-h light environments with a suitable temperature and an adequate supply of food and water. All of the mice were purchased from the Experimental Animal Science and Technology Center of Jiangxi University of Traditional Chinese Medicine ; GAPDH was used as the reference gene  and cDNA was synthesized using a PrimeScript RT reagent Kit with gDNA Eraser , according to the manufacturer’s instructions. The mean values were used for the determination of the mRNA expression levels using the comparative ΔCq (ΔCt) method, according to the equation 2nce gene . All priv/v) NP-40). In addition, PR-619 (50 μM) was used during the extraction of ubiquitin. Equal amounts (30 μg) of protein were separated by 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and then electrophoretically transferred onto the PVDF membranes  and processed per the antibody manufacturer’s instructions. After incubating with the primary antibodies against PTMs, the filter was visualized using anECL detection kit . Analysis of PTMs in the different stages of female mice was conducted by quantifying the grey value of target bands detected by the corresponding antibodies normalized to those detected by GAPDH for anti-pan-PTM . Histones were extracted from the different stages of mice using NETN Lysis Buffer , 0.5 mM EDTA, and 0.5%  containing PMSF  and cocktail, the cell lysates were precleaned with protein G agarose  at 4 °C for 1 h, after which the supernatant was incubated with the indicated antibodies and protein G agarose beads at 4 °C overnight. On the second day, immunocomplexes combined with beads were washed with lysis buffer, followed by western blot.p < 0.05 was considered significant; p < 0.01 and p < 0.001 were considered extremely significant.All analyses were performed using GraphPad Prism 5.01 . The statistical comparisons among different groups were expressed as the mean ± standard error (SEM) and the data from the experiments were analyzed by ANOVA. The threshold of


L375 did not significantly cleave mRNAs containing an unmethylated 5’GpppN cap, indicating that this enzyme specifically hydrolyzes methylated-capped transcripts. A point mutation in the L375 Nudix motif completely eliminated cap hydrolysis, showing that decapping activity is dependent on this motif. Addition of uncapped RNA significantly reduced L375 decapping activity, suggesting that L375 may recognize its substrate through interaction with the RNA body.The giant Mimivirus is a member of the nucleocytoplasmic large DNA viruses (NCLDV), a group of diverse viruses that contain double-stranded DNA (dsDNA) genomes that replicate primarily in eukaryotic hosts. Two members of the NCLDV, Vaccinia Virus (VACV) and African Swine Fever Virus (ASFV), both synthesize Nudix enzymes that have been shown to decap mRNA, a process thought to accelerate viral and host mRNA turnover and promote the shutoff of host protein synthesis. Mimivirus encodes two Nudix enzymes in its genome, denoted as L375 and L534. Importantly, L375 exhibits sequence similarity to ASFV-DP and eukaryotic Dcp2, two Nudix enzymes shown to possess mRNA decapping activity. In this work, we demonstrate that recombinant Mimivirus L375 cleaves the 5’ m Acanthamoeba species, possesses a 1.2 Mb genome encoding over 900 proteins  GTP, and 0.1 mM S-adenosylmethionine . The mutated L375(E258Q) protein was expressed in Escherichia coli and then purified by affinity chromatography concurrently with the wild-type recombinant proteins , as well as their presence in some of the non-living viruses. Three NCLDV Nudix hydrolases have been characterized to date, VACV D9, VACV D10, and ASFV-DP. Each of these enzymes have been shown to possess intrinsic mRNA decapping activity in vitro –15. Impoin vitro ,21–26.7GDP; a reaction that is dependent on an intact Nudix box. Like L375, VACV D9 was shown to preferentially hydrolyze methylated rather than unmethylated mRNA caps [Here we show that the Mimivirus Nudix enzyme L375 hydrolyzed the mRNA cap to release mRNA caps . In contRNA caps ,20. EukaRNA caps –25,27,32Similar to the viral and eukaryotic mRNA decapping enzymes characterized to date, uncapped competitor RNA reduced L375 decapping activity, suggesting that L375 also uses the RNA moiety for substrate selection ,25,27,33in vivo experiments confirmed that ASFV interacts with viral and host mRNAs during infection, a property dependent on the N-terminus of the protein, rather than the Nudix motif [The strong affinity of ASFV-DP for RNA was confirmed through an electrophoretic mobility shift assay, in which the migration of uncapped RNA was impeded with the addition of ASFV-DP . Subsequix motif . ASFV-DPix motif . For exaix motif ,35.7GTP, followed by m7GDP, whereas the effect of m7GpppG was negligible (m7GTP>m7GDP>m7GpppG). In comparison, the decapping activity of both VACV D9 and D10 was diminished by all three methylated cap derivatives, with the most striking reduction also observed with m7GTP [As observed for VACV D9 and D10 (but not for ASFV-DP), addition of methylated cap derivatives reduced L375 decapping activity, suggesting that L375 may also recognize the cap structure in addition to the RNA body ,14. Specth m7GTP ,14. Inteth m7GTP ,14.in vitro, suggesting that for ASFV-DP, the RNA body is critical during substrate identification [In contrast, ASFV-DP was not inhibited by addition of methylated cap derivatives, nor could it cleave free methylated cap analog fication ,20. Likefication –25,27. Hin vivo studies characterizing VACV D9, D10, and ASFV-DP. Increased expression of VACV D9, VACV D10, or ASFV-DP resulted in enhanced mRNA turnover of capped viral and host transcripts, further validating that these enzymes decap mRNA to elicit subsequent mRNA degradation [The purpose of mRNA decapping during viral infection has been revealed through radation ,21. Interadation ,35.in vitro and in vivo, suggesting that these two enzymes work together synergistically [In complementary genetic studies in VACV, deletion or inactivation of D10 resulted in persistence of viral and host mRNAs, a delay in the shutoff of host protein synthesis, and a modest reduction of virulence in animal hosts, whereas deletion of D9 did not produce any noticeable defects –19,36. Wstically . The substically . These rstically . More restically .Like VACV and ASFV, Mimivirus encodes its own mRNA capping enzyme and directs a sequential cascade of viral gene expression consisting of early, intermediate, and late phases ,40. DecaAcanthamoeba species, and therefore does not require L375 mRNA decapping activity to evade the complex vertebrate immune systems. However, Mimivirus could still use selective mRNA decapping and degradation of specific host mRNAs to promote a robust Acanthamoeba infection. Future in vivo studies will help elucidate the role of L375 mRNA decapping and decay during Mimivirus infection.In contrast to the animal hosts of VACV and ASFV, Mimivirus primarily infects unicellular S1 Raw images(PDF)Click here for additional data file.


Asthma is a common chronic respiratory disease in children, seriously affecting children's health and growth. This bioinformatics study aimed to identify potential RNA candidates closely associated with childhood asthma development within current gene databases.  GSE65204 and GSE19187 datasets were screened and downloaded from the NCBI GEO database. Differentially expressed long noncoding RNAs (DE-lncRNAs) and mRNAs (DE-mRNAs) were identified using the Bioconductor limma package in R, and these DE-mRNAs were used to perform biological process (BP) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses. Thereafter, weighted gene coexpression network analysis (WGCNA) was utilized to screen the modules directly related to childhood asthma, and a coexpression network of DE-lncRNAs and DE-mRNAs was built. Finally, principal component analysis (PCA) was performed.  In total, 7 DE-lncRNAs and 1060 DE-mRNAs, as well as 7 DE-lncRNAs and 1027 DE-mRNAs, were identified in GSE65204 and GSE19187, respectively. After comparison, 336 overlapping genes had the same trend of expression, including 2 overlapped DE-lncRNAs and 334 overlapped DE-mRNAs. These overlapped DE-mRNAs were enriched in 28 BP and 12 KEGG pathways. Eleven modules were obtained in GSE65204, and it was found that the purple, black, and yellow modules were significantly positively correlated with asthma development. Subsequently, a coexpression network including 63 DE-mRNAs and 2 DE-lncRNAs was built, and five KEGG pathways, containing 8 genes, were found to be directly associated with childhood asthma. The PCA further verified these results. LINC01559 and SNHG8 and mRNAs VWF, LAMB3, LAMA4, CAV1, ALDH1A3, SMOX, GNG4, and PPARG were identified as biomarkers associated with the progression of childhood asthma. LncRNAs  It is caused by a combination of genetic, environmental, and lifestyle factors and is characterized by inflammation, airway hyperresponsiveness, and variable airflow obstruction . Asthma d asthma . Howevers asthma . HoweverMEG3 was highly expressed in healthy tissues adjacent to gastric cancer tissues and that its overexpression could inhibit the proliferation and metastasis of gastric cancer cells through the p53 signaling pathway. Cao et al. [RMRP was significantly upregulated in bladder cancer tissues and was closely related to tumor size, lymph node metastasis, and survival time of patients. LncRNA RMRP can promote the proliferation, migration, and invasion of bladder cancer cell lines [Long noncoding RNAs (lncRNAs) are a class of noncoding RNAs with a length greater than 200 nt and are regulators of various biological processes, including chromatin structure changes, transcriptional activation/inhibition, intracellular trafficking, and microRNA (miRNA) chelation and translation efficiency regulation , 7. Altho et al.  reportedll lines . All thell lines  analyzedll lines . NeverthGAS5, HCP5, PART1, SNHG11, and SNHG5) to predict the prognosis of ovarian cancer. Therefore, this study aims to identify differentially expressed RNAs (DERs) closely related to asthma progression in children using WGCNA. These findings may provide novel diagnostic biomarkers and therapeutic targets for the development of childhood asthma.Weighted gene coexpression network analysis (WGCNA) has been successfully used to identify highly correlated gene clusters (coexpression modules) and to identify potential biomarkers or therapeutic targets significantly associated with clinical phenotypes , 16. Zhahttps://www.ncbi.nlm.nih.gov/) [On November 5, 2020, datasets that met the following criteria were screened in the NCBI GEO database (ih.gov/)  with theih.gov/)  and GSE1https://asia.ensembl.org/index.html), including annotation files, transcript IDs, and RefSeq IDs. The expression profiles in GSE65204 and GSE19187 were reannotated into mRNAs and lncRNAs.The detailed annotation information of the platforms was downloaded from Ensembl genome browser, release 96 (https://bioconductor.org/packages/release/bioc/html/limma.html) compiled in R 3.6.1 [2 fold change (FC) |>0.5. By comparing DERs between GSE65204 and GSE19187, the DERs with the same differential expression were chosen as the overlapped DERs (overlapped DE-lncRNAs and DE-mRNAs) related to asthma. These screened overlapped DE-mRNAs were utilized to perform biological process of Gene Ontology (GO BP) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses using DAVID version 6.8 (https://david.ncifcrf.gov/) [P value of <0.05 was used as the cutoff criterion for enrichment significance.The Bioconductor limma package  . A P valhttps://cran.r-project.org/web/packages/WGCNA/index.html) compiled in R [Pp value of the DE-mRNAs were calculated using a hypergeometric algorithm [P < 0.05 and fold enrichment >1 served as the thresholds for module filtering. The formula of the hypergeometric algorithm was (kNMn)=C (kM) × C ÷ C(nN), where N denotes all genes involved in WGCNA analysis, M denotes the number of genes in each module, n denotes the number of overlapping DE-mRNAs, and k represents the number of DE-mRNAs mapped to the corresponding module.The WGCNA package . Subsequently, a coexpression network was constructed, and Cytoscape 3.6.1 (https://www.cytoscape.org/) was used for visualization [P < 0.05).The Pearson correlation coefficient (PCC) between DE-mRNAs enriched in the modules and overlapped DE-lncRNAs from the earlier differential expression screen was calculated using the cor function in R (lization . Afterwahttps://ctd.mdibl.org/) [With “asthma” as the keyword, Comparative Toxicogenomics Database (2019 update) (bl.org/)  was utilhttps://CRAN.R-project.org/package=psych) compiled in R.PCA is a dimension reduction technique that transforms multiple variables into a few principal components that can reflect most of the information of the original variables . PCA wasSNHG8 and LINC01559) and 334 overlapped DE-mRNAs.After reannotation and processing, 135 lncRNAs and 11,808 mRNAs were obtained. Afterwards, DEGs between normal samples and asthma samples were identified. In total, there were 1067 DERs (7 DE-lncRNAs and 1060 DE-mRNAs) found in GSE65204 and 1034 DERs (7 DE-lncRNAs and 1027 DE-mRNAs) found in GSE19187 . By compThese 334 overlapped DE-mRNAs were used for GO BP and KEGG analyses. It was found that these DE-mRNAs were enriched in 28 biological processes, such as “negative regulation of endopeptidase activity,” “oxidation-reduction process,” “epidermis development,” “T-cell costimulation,” “O-glycan processing,” “protein N-linked glycosylation via asparagine,” “glutathione derivative biosynthetic process,” “extracellular matrix organization,” and “bone coagulation” . AdditioWGCNA was performed on all DERs in GSE65204 and GSE19187, using GSE65204 as the training set and GSE19187 as the validation set. To ensure that the gene expression levels in each dataset were comparable, we first analyzed the expression level and connectivity consistency of all gene expression values detected in these two datasets. It was found that the gene expression levels and network connections were significantly positively correlated between the training and validation sets, which indicated that the two datasets were indeed comparable . We subsZ-score higher than 5 , laminin subunit beta 3 (LAMB3), and laminin subunit alpha 4 (LAMA4) genes were involved in “ECM-receptor interaction,” “focal adhesion,” and “PI3K-Akt signaling pathway,” while LAMB3 and LAMA4 were also involved in “pathways in cancer.” In addition, caveolin-1 (CAV1) was involved in “focal adhesion,” and aldehyde dehydrogenase 1 family member A3 (ALDH1A3) and spermine oxidase (SMOX) were associated with “beta-alanine metabolism.” G protein subunit gamma 4 (GNG4) was related to the “PI3K-Akt signaling pathway” and “pathways in cancer.” Additionally, peroxisome proliferator-activated receptor gamma (PPARG) was involved in “pathways in cancer.” The coexpression network found that lncRNA LINC01559 may be coexpressed with VWF, LAMB3, CAV1, ALDH1A3, SMOX, and GNG4. lncRNA SNHG8 might coexpress with VWF, LAMB3, LAMA4, ALDH1A3, SMOX, and PPARG. This analysis suggests that these lncRNAs, mRNAs, and pathways may be closely associated with childhood asthma development.A total of 195 KEGG pathways directly related to asthma were obtained from the CTD database. After comparison with the DE-mRNA-enriched pathways in the coexpression network, the five enriched KEGG pathways (containing eight genes) were shown to be directly associated with asthma . It is cVWF, LAMB3, LAMA4, CAV1, ALDH1A3, SMOX, GNG4, and PPARG are closely associated with the occurrence and development of childhood asthma.PCA was performed on the eight genes involved in the five asthma-related pathways. Through the calculation and analysis of gene expression values in GSE65204 and GSE19187, a total of eight principal components (PCs) were obtained by fitting. Among them, the cumulative contribution rate of PC1, PC2, and PC3 was over 80%, which implied that these three PC factors contained significant information about the original variables . Thereafter, PC1, PC2, and PC3 were used to construct three-dimensional diagrams of the sample distribution and to build receiver operating characteristic (ROC) curves. As shown in Figures SNHG8 and LINC01559) and 334 overlapped DE-mRNAs had the same differential expression. After that, significantly stable modules related to asthma were identified by WGCNA, and a coexpression network was built with 2 DE-lncRNAs and 63 DE-mRNAs. After functional analysis of genes in the coexpression network, five KEGG pathways  contained 8 genes  which were found to be directly related to childhood asthma development and have potential for use as disease progression biomarkers.Asthma is a common chronic respiratory disease in children, seriously affecting children's health and growth . Since tSNHG8 and LINC01559 were found to be associated with the progression of childhood asthma. SNHG8, located on 4q26, regulates tumorigenesis and metastasis and controls the progression of multifarious diseases, such as hepatocellular carcinoma [LINC01559 was upregulated in gastric cancer tissues and could stimulate the PI3K-Akt signaling pathway to accelerate the progression of gastric cancer [SNHG8 and LINC01559 may participate in the occurrence and development of childhood asthma by regulating cell proliferation, migration, and PI3K-Akt signaling pathway.Previous studies have shown that lncRNAs play important roles in the development of pediatric airway diseases, including asthma , 30. Ourarcinoma , pancreaarcinoma , endometarcinoma , and gasarcinoma . Wang etc cancer . TherefoLINC01559 might coexpress with VWF, LAMB3, CAV1, ALDH1A3, SMOX, and GNG4, and SNHG8 might coexpress with VWF, LAMB3, LAMA4, ALDH1A3, SMOX, and PPARG, which were enriched in five KEGG pathways, including “ECM-receptor interaction,” “focal adhesion,” “beta-alanine metabolism,” “PI3K-Akt signaling pathway,” and “pathways in cancer.” PCA further verified that these eight genes are important in childhood asthma. VWF, a large plasma glycoprotein, plays a vital role in mediating the balance between blood clotting and bleeding [VWF was highly expressed in an IL-25-induced murine asthma model and was associated with bronchial mucosal vascular remodeling in asthma. LAMB3 and LAMA4 belong to the laminin subunit family and have been reported to be involved in the metastasis and invasion of some types of cancer [LAMB3 could mediate apoptosis, proliferation, migration, and invasion of pancreatic ductal adenocarcinoma cells through the PI3K/Akt signaling pathway. Another study found that high expression of LAMA4 may be a new marker of tumor invasion and metastasis in human hepatocellular carcinoma [CAV1 is a carcinogenic membrane protein associated with extracellular matrix tissue, cell migration, cholesterol distribution, endocytosis, and signal transduction [CAV1 participated in focal adhesion, thus contributing to the pathogenesis of childhood asthma. GNG4 is one of the fourteen γ-subunit proteins of the G protein trimer complex [GNG4 associated with the PI3K-Akt signaling pathway to participate in rectal cancer through using a bioinformatics approach. PPARG is one of the three subtypes of peroxisome proliferator-activated receptors, and its activation has been reported to regulate the synthesis and release of immunomodulatory cytokines from various cell types [In addition, from the lncRNA-mRNA coexpression network, bleeding . A studybleeding  showed tf cancer , 39. A sf cancer  demonstrarcinoma . CAV1 issduction . In our  complex , and Liu complex  found thll types . A previll types .ALDH1A3 and SMOX were also essential for childhood asthma and were enriched in the “beta-alanine metabolism” pathway. ALDH1A3 is widely distributed in normal tissues and abnormally expressed in a variety of cancers [ALDH1A3 can serve as an activator of mesenchymal differentiation and may be a marker for predicting the survival rate of glioblastoma patients. SMOX, a member of the polyamine oxidases, catalyzes the oxidative degradation of polyamine spermidine to produce spermidine and is caused by a variety of stimuli, including bacterial infection and oxidative stress [SMOX in bronchial epithelial cells (BECs) of asthmatic lung samples, and SMOX knockdown resulted in asthma in naive mice, such as airway hyperresponsiveness, remodeling, and BEC apoptosis. Combined with our results, we speculate that lncRNAs LINC01559 and SNHG8 may be coexpressed with VWF, LAMB3, LAMA4, CAV1, ALDH1A3, SMOX, GNG4, and PPARG, and these lncRNAs and mRNAs may be direct biomarkers related to the occurrence and progression of childhood asthma. However, the relationships between lncRNAs and their target genes need to be further characterized, and use of these identified lncRNAs and mRNAs as biomarkers will require further validation in a clinical setting.Additionally,  cancers . Li et a cancers  indicatee stress . Jain ete stress  found loLINC01559 and SNHG8 may be coexpressed with VWF, LAMB3, LAMA4, CAV1, ALDH1A3, SMOX, GNG4, and PPARG. Additionally, these lncRNAs and mRNAs may be directly involved in the pathogenesis of childhood asthma through ECM-receptor interaction, focal adhesion, beta-alanine metabolism, PI3K-Akt signaling pathway, and pathways in cancer. The two lncRNAs and eight genes could explain potential, novel, pathological, and molecular mechanisms for childhood asthma and may serve as candidates for therapeutic strategies for this disease.In conclusion,


Non-human primates (NHP) are an important source of viruses that can spillover to humans and, after adaptation, spread through the host population. Whereas HIV-1 and HTLV-1 emerged as retroviral pathogens in humans, a unique class of retroviruses called foamy viruses (FV) with zoonotic potential are occasionally detected in bushmeat hunters or zookeepers. Various FVs are endemic in numerous mammalian natural hosts, such as primates, felines, bovines, and equines, and other animals, but not in humans. They are apathogenic, and significant differences exist between the viral life cycles of FV and other retroviruses. Importantly, FVs replicate in the presence of many well-defined retroviral restriction factors such as TRIM5α, BST2 (Tetherin), MX2, and APOBEC3 (A3). While the interaction of A3s with HIV-1 is well studied, the escape mechanisms of FVs from restriction by A3 is much less explored. Here we review the current knowledge of FV biology, host restriction factors, and FV–host interactions with an emphasis on the consequences of FV regulatory protein Bet binding to A3s and outline crucial open questions for future studies. Pan troglodytes schweinfurthii, whose natural habitat includes Kenya. Since the original HFV isolate came from a person who might have had contact with chimpanzees in Kenya, the virus was probably acquired as a zoonotic infection . For detailed reviews on foamy virus epidemiology and zoonotic infections see [The first description of a foamy virus (FV) was reported in 1954 . In 1971ions see ,7. HFV hions see ,11,12. Nions see ,16,17,18−8 base substitutions per site per year, making FV the slowest-evolving virus documented so far. These investigations moreover revealed highly congruent relationships, indicating virus-host co-evolution for at least 30–40 million years [FV genomes display high evolutionary conservation among all the species infected, and FV genetic variability within one infected animal is very low over time (<1% variation) . The phyon years ,21. The on years ,6,22,23.on years ,24.Macaca fascicularis) population and Wild red colobus monkeys in Tai national park, West Africa [SFVs are highly prevalent. In captive primate populations, infection rates ranging from 70% to 100% are reported in adult animals ,6,23, ant Africa ,25 as wet Africa . A recent Africa . Adult ct Africa . Hunterst Africa ,29,30,31t Africa ,33,34, at Africa ,36.Rhinolophus affinis) [FVs are also prevalent in non-simian hosts such as cats ,38, pumaaffinis) , and likaffinis) ,45,46,47affinis) ,49,50. Iaffinis) . These rDaubentonia madagascariensis, a strepsirrhine primate), the two-toed (Choloepus hoffmanni) and three-toed sloths (Bradypus pygmaeus) from South America, platyfish (Xiphophorus maculatus), and cod species [Latimeria chalumnae) [When retroviruses get access to gametocytes, the resulting endogenous provirus can be passaged in the germline along with the host cell genome, but in most cases carries inactivating mutations or deletions ,53. Endo species ,55,56, aalumnae) . Very realumnae) . Recent alumnae) ,60,61,62alumnae) .Foamy viruses have complex RNA genomes , and encapsidation of Pol is thought to be mediated by viral RNA bridging Gag and Pol molecules ,74,75,76In vitro, FVs have the capacity to infect most cell types of vertebrate origin from fish to humans [FVs are unable to establish a productive infection in G1/S growth-arrested or nondividing cells , requirio humans ,97,98. Ho humans ,100. Glyo humans ,103,104,o humans ,106. Wheo humans ,109,110.o humans .Like other viruses, FVs not only have to exploit various host machinery for their productive replication, but they have also to escape or counteract host antiviral responses, such as innate immune sensing and inhibition by cellular restriction factors. This area of research is particularly interesting because FV infections are apparently apathogenic in the hosts, but in vitro, FV infection triggers cytopathic effects and ultimately leads to cell death . HoweverInnate sensing of FV is not well characterized. Early studies suggested a lack of type I interferon (IFN-I) induction by simian and human FV infections in different cell lines ,116,117.Mus caroli, was found to inhibit FFV [Specific IFN-induced cellular gene products are able to restrict different retroviruses, including FVs ,141,142.ibit FFV ,144. Humibit FFV . A recenibit FFV . Interesibit FFV . Moreoveibit FFV  and humaAnother type of innate frontline defense against retroviruses is mediated by the members of the APOBEC3 (A3) cytidine deaminases family. Our earlier study reported the DNA editing of the FFV genome in A3-positive feline CRFK cells (non-permissive phenotype), which dramatically diminished the FFV titer . Two othA3, mA3 for instance) genes, humans express seven A3 proteins, and bats likely up to 18 distinct A3s from as many genes [The apolipoprotein B mRNA-editing enzyme, catalytic polypeptide-like 3  family of single-stranded (ss) DNA cytidine deaminases provides mammals with an innate immune barrier against retroviruses, retrotransposons, and other viral pathogens ,152,153.ny genes ,155,156.ny genes ,157,158.The domain structure of A3s consists of a conserved sequence of characteristic motifs (α1-β1-β2/2′-α2-β3-α3-β4-α4-β5-α5-α6) ,152. At gag gene that is initiated by an upstream CUG codon (p80)), can antagonize mA3 and certain human A3s’ restriction activity [During infection by lentiviruses (a group of retroviruses that includes HIV-1) the antiviral activity of the host-species’ A3 is counteracted by the viral Vif protein. Vif directly binds to A3 and recruits an E3 ubiquitin ligase complex for polyubiquitination and proteasomal degradation of A3 ,172,173.activity ,181,182.activity ). Along activity ,184,185.FVs encode two non-structural proteins, Tas and Bet, which are implicated in overcoming host innate immunity. Whereas the transcriptional activator function of Tas and its role in inhibiting RNA silencing pathways have been known for some time , a dedicThe extent of protein sequence homology of Bet is low among FVs ,48,89. Tab initio protein structure prediction [ab initio based on the full sequences with ROSETTA3 [Despite recent advances in ediction , templatediction . Howeverediction ,208,209,ediction ,211,212,ediction , or SWISediction ,215 yielROSETTA3  or the RROSETTA3  did not ROSETTA3 ,219,220 ROSETTA3  searchesα atom RMSD between the generated models and the corresponding ab initio modeled domain of PFV Bet. This consensus approach was also validated by TopScore [Therefore, we decided to predict domain boundaries within the sequence of the SFV-chimpanzee Bet protein sequence with TopDomain  Figure A and subTopScore  in that ASH1 mRNA transportation complex [For the N-terminal domain, the DALI web server  identifi complex  -mediated retroviral inhibition . FFV perComputational analysis of Bet sequences from BFV, EFV, SFV, FFV, PFV, and SloEFV identified six conserved motifs in the Bel2 domain of Bet . The entNotably, two earlier studies using the PFV-based viral vector system reported controversial data regarding the function of Bet as an A3G antagonist ,136. RusTo learn more about mechanistic aspects and functional consequences of Bet binding to A3s, our lab has conducted two independent studies involving A3C and A3G. Both have reported a direct physical interaction of Bet with A3C and A3G, independent of RNA, which traps the A3s in the cytoplasm rendering them unavailable for incorporation into progeny virions ,184. TheAdditionally, our recent study suggested that Bet can counteract A3B without decreasing the steady-state level of A3B . IntriguSeveral investigations identified Bet as a countermeasure to protect viruses from A3s by blocking packaging into progeny virions, a function reminiscent of lentiviral Vif proteins. The reverse transcription complex of FV is not well described, and it is unknown how the antiviral A3 proteins get co-packaged in such complexes. Additional work focusing on A3 interactions with different FV Gag, viral genomic RNA, and DNA is needed to confirm the role of A3 in producer and target cells using both authentic foamy-viral as well as lentiviral systems.There has been impressive progress in biochemical and structural understanding of A3 and HIV-1 Vif, and molecular studies of purified Bet proteins remain for future work. Likewise, A3 binding motifs of Bet were not comprehensively studied; while the physical interaction of Bet-A3 and Bet binding regions on A3s were demonstrated, we do not know much about the key regions of Bet involved in these interactions. Indeed, structural investigations on Bet will be important, but are complicated by its folding/stability issues . Whereas it was demonstrated that Bet impairs the cytosolic solubility of A3s and drags A3s into insoluble complexes in the cell, a direct/indirect function of Bet in shielding viral DNA during RT (late) from A3s in target cells has not been proposed. Additionally, nuclear-localized A3A, A3B, and A3C may impede Gag tethering, genome integration, and transcription of Tas/Bet or LTR-driven viral genes. These open questions and the discovery of currently unknown functions of Bet identification await future research.