Uterine sarcomas are rare tumors accounting for 3,4% of all uterine cancers. Even after radical hysterectomy, most patients relapse or present with distant metastases. The very limited clinical benefit of adjuvant cytotoxic treatments is reflected by high mortality rates, emphasizing the need for new treatment strategies. This review summarizes rising potential targets in four distinct subtypes of uterine sarcomas: leiomyosarcoma, low-grade and high-grade endometrial stromal sarcoma, and undifferentiated uterine sarcoma. Based on clinical reports, promising approaches for uterine leiomyosarcoma patients include inhibition of VEGF and mTOR signaling, preferably in combination with other targeted or cytotoxic compounds. Currently, the only targeted therapy approved in leiomyosarcoma patients is pazopanib, a multitargeted inhibitor blocking VEGFR, PDGFR, FGFR, and c-KIT. Additionally, preclinical evidence suggests effect of the inhibition of histone deacetylases, tyrosine kinase receptors, and the mitotic checkpoint protein aurora kinase A. In low-grade endometrial stromal sarcomas, antihormonal therapies including aromatase inhibitors and progestins have proven activity. Other potential targets are PDGFR, VEGFR, and histone deacetylases. In high-grade ESS that carry the YWHAE/FAM22A/B fusion gene, the generated 14-3-3 oncoprotein is a putative target, next to c-KIT and the Wnt pathway. The observation of heterogeneity within uterine sarcoma subtypes warrants a personalized treatment approach. Although uterine sarcomas only account for 3,4% of all uterine corpus malignancies, they entail a high mortality rate , 2. RepoNCT01533207). Another approach in advanced disease is trabectedin, a marine-derived drug that has shown minor first-line and second-line activity in LMS patients, but is currently not approved by the Food and Drug Administration (FDA) [Uterine leiomyosarcomas, arising from the myometrium, are generally high-grade tumors accounting for 60% of all uterine sarcomas . Due to on (FDA) , 12, 13.Uterine LMS show multiple and varied genetic aberrations and very complex, often aneuploid or polyploid, karyotypes , 15. ThiPDGF) and its receptor PDGFR have been implicated in the pathophysiology of multiple tumor types including gastrointestinal stromal tumor (GIST), glioblastoma, and dermatofibrosarcoma protuberans [β amplifications in uLMS [β [α, while no activating mutations have been found in the gene [Mutations in receptor tyrosine kinases (RTK), leading to aberrant pathway activation, have often been reported in cancer. Amplifications, mutations, and rearrangements of platelet-derived growth factor  were scored positive for VEGF [ VEGFR1 and VEGFR2 are frequently expressed in uLMS [ in vitro and in vivo uLMS models after treatment with vandetanib, a VEGFR2/epidermal growth factor receptor (EGFR) inhibitor [γ-secretase , reported prolonged stable disease in one of three LMS patients  [Althoughfor VEGF –32. In a in uLMS , 31. In  in uLMS . Remarka in uLMS . Furthernhibitor . Moreoveecified) . In contecified) . ERBB2/HER-2, belonging to the ERBB family of tyrosine kinase receptors, is of great clinical importance in breast cancer, where it is tackled by the monoclonal antibody trastuzumab [Overexpression ofstuzumab . It has stuzumab . Three sstuzumab –41. In tstuzumab –41. Henc EGFR. EGFR has been shown to be upregulated in uLMS when compared to normal controls [ in vitro as well as in an in vivo xenograft model [Another important cell growth regulator of the ERBB family iscontrols . One stucontrols . Two othcontrols , 31. A rcontrols . Interesft model .IGF2), which activates IGF 1/2 receptors (IGF1/2R), has been reported to be upregulated in uLMS [Insulin-like growth factor 2  and its tyrosine kinase receptor, neurotrophic tyrosine kinase receptor type 2 , have recently been reported to be upregulated in uLMS compared to LM and myometrium. Treatment of MES-SA cells, which are derived from uterine sarcoma, with the multikinase inhibitor K252a or the NTRK2 ectodomain suppressed proliferation and induced apoptosis. Moreover, in MES-SA-injected mouse models, administration of K252a resulted in smaller tumors, lower proliferation rates, and more apoptosis [Lastly, brain-derived neurotrophic factor (poptosis .PIK3/AKT/mTOR) pathway has been implicated in uLMS and may play a role in its etiology  and on the membrane (low expression in 80% and high expression in 20% of samples) [β-catenin pathway is targetable through many different pathway players with commercially available inhibitors, but this approach has not yet been tested in uLMS [Next to the mTOR pathway, in uLMS . The Wntcontrols . The autsamples) . Other gsamples) , 64. The in uLMS .ROR2), which is involved in noncanonical Wnt signaling TGF-β/BMP) signaling may play a role in uLMS  was found to be highly overexpressed. Of note, almost all genes with >9-fold increase in expression were involved in regulating chromosomal homeostasis and spindle assembly, suggesting that proteins involved in these functions could be useful therapeutic targets. Indeed, the single targeting of AURKA with siRNA or MK-5108 inhibited uLMS cell proliferation in vitro and decreased the number and size of tumor implants in vivo [In a recent genome-wide study of 12 uLMS, aurora kinase A ( in vivo . Similar in vivo . Of note in vivo .MDM2 is an oncogene that negatively regulates p53 function by three mechanisms: (1) targeting p53 for ubiquitin-based degradation, (2) blocking the p53 transcriptional activation domain, and (3) shuttling p53 from the nucleus to the cytoplasm [ in vitro in 40 cell lines. Synergy was observed upon simultaneous inhibition of MDM2 and MEK and/or PI3K. Interestingly, this effect was not dependent on the mutation status of genes in the PI3K pathway, and the highest inhibitory effect was noted when all three molecules were blocked [ytoplasm . Blockinytoplasm , 72, 73.ytoplasm , 75. MDMytoplasm –78. To t blocked .HDAC) inhibitors, as detected by a tremendous increase in apoptosis and decrease in cell proliferation [ in vitro [ in vivo [Furthermore, MDM2 inhibitors were found to greatly synergize with histone deacetylase  and/or progesterone receptors (PR), , 87–90. In uLMS, recurrent regions of loss often include tumor suppressor genes (TSG) such as phosphatase and tensin homolog (PTEN), tumor protein p53 (TP53), retinoblastoma 1 (RB1), and cyclin-dependent kinase inhibitor 2A (CDKN2A) , 92–94.CDKN2A encodes the p16 protein, which controls cell proliferation by inhibiting cell cycle progression. When p16 is absent, cyclin-dependent kinases bind to cyclins, enabling them to phosphorylate RB1. Upon phosphorylation, RB1 releases the transcription factor E2F, stimulating cell cycle progression [gression . In uLMSgression . RB1 gene is frequently deleted in uLMS [ RB1 gene is named after the corresponding cancer type hereditary retinoblastoma, where the gene is homozygously deleted. It was shown recently that retinoblastoma patients with the hereditary type (RB1 deletion) have an increased risk of uLMS; 3,2% of patients developed uLMS, which corresponds to an excess risk of 3,9/10000 women [Further, the in uLMS , 93. The00 women .PTEN is a negative regulator of the PI3K-AKT-mTOR pathway, and loss of PTEN is associated with increased pathway activity, as found in many cancers , and TP53 accelerated tumor progression in this mouse model. Further, the authors described downregulation of BRCA1 in 29% of uLMS, most likely due to promoter methylation [Another tumor suppressor gene, which is mutated in approximately 50% of cancers and frequently deleted in uLMS, is TP53 , 93, 101rphology . Additiohylation .Tumor cells that display loss of tumor suppressor function can be tackled by the inhibition of another protein that has become indispensable after loss of the first tumor suppressor, that is, the synthetic lethality principle . For exaLow-grade endometrial stromal sarcomas comprise about 20% of uterine sarcomas . They ar PDGF signaling pathway has proven interesting to explore in LGESS. Taking together seven studies, 71/141 (50%) of LGESS cases showed expression of PDGFR-α and 53/127 (42%) of LGESS were positive for PDGFR-β [α and PDGFR-β, designating this receptor as a potential therapeutic target in LGESS and warranting further research [ c-KIT expression in 52 cases and summarized previous studies, revealing only 16 c-KIT positive cases in 203 included LGESS (8% positive), suggesting c-KIT is not a valuable target in LGESS [From IHC studies, the PDGFR-β , 110–115 PDGFR-β , 117. Onin LGESS . The onlin LGESS , 117. EGFR expression has been detected in 35 cases (22%), with substantial differences between studies, varying from 0/39 to 14/20 positive cases [Further, in a total of 156 LGESS samples,ve cases , 119. Heve cases . No stud VEGF [γ-secretase inhibitor, reported partial response in a LGESS patient [Combining three studies, with a total of 22 LGESS patients, 60% of tumors expressed VEGF , 118. VE VEGF . A recen patient . This pr HDAC2 has been reported. Although treatment of the ESS-1 cell line with a HDAC inhibitor resulted in cell cycle arrest and cell differentiation in vitro, the results failed to translate into clinical response: in a phase II trial exploring the effect of the HDAC inhibitor panobinostat, no responses were observed in 3 LGESS patients [Histone deacetylases and the inhibition thereof are under investigation in many cancer types. In LGESS, overexpression ofpatients , 122. Onpatients , 123. Inpatients . One reppatients . Wnt/β-catenin pathway in endometrial stromal tumors. Taking together results from 4 independent studies, 75/121 LGESS show positive β-catenin-staining [β-catenin that allows cell cycle progression and contributes to cell proliferation, is only rarely detected in LGESS [Also little is known on thestaining , 125–127in LGESS , 128.Hormone receptors are expressed in 70–80% of LGESS, with proven therapeutic importance for over a decade [a decade , 129–132a decade –137. Thea decade . Of notea decade , 133. HoHigh-grade ESS (HGESS) arise from the endometrial stroma, but unlike LGESS, they show a high-grade round-cell morphology and are clinically more aggressive , 139. Th 14-3-3 oncoprotein [Recently, the translocation t has been described in HGESS , 141–145oprotein . PDGFR signaling may be involved in HGESS and UUS. Both PDGFR-α and PDGFR-β have been detected in 37% of 30 reported samples, warranting studies on its clinical relevance [Similar to LGESS,elevance , 115. ERBB2 in HGESS/UUS. In a single study by Amant et al., one of four (25%) cases showed amplification and overexpression in the primary and the recurrent tumor [ EGFR overexpression has been reported more frequently in HGESS/UUS. Taken together, 16/33 (48%) cases were EGFR-positive by IHC [ c-KIT overexpression [NCT01979393). Cabozantinib is a multikinase inhibitor targeting VEGFR2, c-Met, Ret, Kit, Flt-1/3/4, Tie2, and AXL, which has been approved for treatment of progressive metastatic medullary thyroid cancer [Only one group has studiednt tumor . Hence, e by IHC , 119. A e by IHC . Anotherpression . C-KIT wpression . Lastly,d cancer . In the Cyclin D1 expression has been detected exclusively in UES-U, whereas all reported UES-P cases were negative [ Wnt/β-catenin pathway (β-catenin has been demonstrated in 6/7 (85%) UES-U cases and only in 2/6 (33%) UES-P cases [β-catenin has been reported in 9/12 additional HGESS/UUS cases, but these were not further classified [β-catenin pathway are available, it may be valuable to test this approach in translocation-related HGESS cases.negative , 146. Cynegative . Hence,  pathway . Indeed,-P cases . Expressassified , 151. As sex hormone receptors. Kurihara et al. found that ER and PR are present in at least half of UES-U cases, while they were not detected in any UES-P cases [UES-U and UES-P also differ in the expression of-P cases . The pre-P cases . A summaIn this review, we summarize the latest reported aberrations with potential therapeutic applicability in uterine leiomyosarcomas, low-grade and high-grade endometrial stromal sarcomas, and undifferentiated uterine sarcomas.Among the scarce clinical reports on targeted treatments in uterine leiomyosarcoma patients, a promising approach involves tackling the PI3K/AKT/mTOR pathway. However, mTOR inhibition leads to activation of AKT through upstream receptor tyrosine kinase signaling, due to interruption of feedback inhibition . Therefoγ-secretase inhibitor or chemotherapeutics such as temozolomide [Another promising approach in uLMS is the interruption of VEGF signaling. Although most responses are published in case reports, sorafenib and aflibercept made it to a phase II trial, resulting in a minor response: stable disease was reached in 4/12 (33%) and 11/41 27%) uLMS patients, respectively , 37. So % uLMS pazolomide –36. in vitro, but combination regimens may be necessary to reach efficacy in vivo.In low-grade endometrial stromal sarcomas, particularly aromatase inhibitors and progestins have proven their effectiveness. Further, PDGF and VEGF signaling seem to be potential targets in LGESS, but at the moment only case reports have been published. HDAC inhibitors have been shown to be effectiveHigh-grade endometrial stromal sarcomas often carry the translocation t, giving rise to the fusion gene YWHAE/FAM22A/B . The resMoreover, Wnt pathway players have been detected specifically in this HGESS subgroup , 146. HeUndifferentiated uterine sarcomas appear to be underexplored, as no targets could be identified from literature. Although partly due to their rarity, the recent reclassification of the tumor subtypes by the World Health Organization also impedes the identification of the UUS subtype in older publications .In conclusion, combinations of treatments and multitargeted compounds such as pazopanib overall generate higher clinical benefit than single agents, partly by tackling escape routes that lead to resistance. While multiple promising targets were identified for uterine leiomyosarcoma and low/high-grade endometrial stromal sarcoma, the lack of studies on undifferentiated sarcomas warrants more multicentric studies on these rare tumors. Since randomized trials are scarce for uterine sarcoma patients, we support a personalized therapy approach. Preclinical studies testing the rational combination of existing inhibitors on cell lines and xenografts might pave the way to use these inhibitors in off-label use or repurposed drugs in uterine sarcoma patients, corresponding to their genetic profile.


The role played by total cholesterol (TC) in risk for subarachnoid hemorrhage (SAH) is unclear because studies report both high and low TC each as a risk factor. We performed a systematic review to clarify associations between lipid profile and SAH.2-statistics.Our literature search comprised Pubmed, Scopus, and Cochrane Library databases with no language, publication year, or study type limitations. The Preferred Reporting Items for Systematic reviews and Meta-analyses (PRISMA) checklist guided our reporting. Data forms adapted from the Critical Appraisal Skills Program (CASP), and Cochrane Collaboration guidelines provided a platform for risk-of-bias evaluation. We used a random effects model to calculate pooled estimates and assessed heterogeneity with I2 = 79.5%, I2 = 89.0%, and I2 = 84.3%) and considerable in prospective (I2 = 46.0%). We therefore focused on qualitative analysis and found that only two studies had a low risk of bias. According to these studies high TC increases risk for SAH in men, whereas the role of HDL remained unclear.Of the final 21 studies reviewed, 12 were prospective and 9 retrospective. All studies assessed TC, four assessed HDL, and none LDL in risk for SAH. Heterogeneity among all, retrospective, and Asian studies was high  of hypercholesterolemia may exceed the PARs of smoking and hypertension in men. Apart from diabetes and obesity, the risk-factor profile of SAH seems to resemble that of other cerebrovascular diseases, at least in men. Epidemiological studies of risk factors of subarachnoid hemorrhage (SAH) are challenging to conduct for a number of reasons, such as relatively young age at diagnosis, low overall incidence of disease, and high death-rate from the first bleed. The latter implies that outside-hospital deaths from SAH are comprehensively diagnosed only in countries with a high coverage of forensic autopsies for sudden deaths. The number of large, long-term, and population-based prospective studies on risk factors for SAH is limited.–3 AccordThe role of total cholesterol (TC) in risk for SAH is conflicting, since studies report both high–6 and loThe protocol of this study is available in PROSPERO  and follows the Preferred Reporting Items for Systematic reviews and Meta-analyses for Protocols Statement (PRISMA-P). The authors J.V.L. and M.K. did all steps of the protocol below; discussion with third-author J.K. resolved disagreements. We performed the literature search in two parts using Cochrane Library, Pubmed, and Scopus databases with no language limitations “We used the PRISMA checklist as a gui2-statistics. For two studies,[f (RR-1)/[pf (RR-1)+1], where pf is population fraction with hypercholesterolemia. Stata statistical software version 12.1 Stata Corp performed all the calculations.Based on the Cox proportional hazards model, we estimated sufficient sample size for prospective follow-up studies. We used a standard significance value of p<0.05, standard power value of P = 0.8, and incidence value of 20/100 000. For optimistic power analysis, we selected the hazard ratio (HR) 2.0 (95% CI 1.0–4.0) as effect size with a wide confidence interval and low correlation factor value of 0.1 between covariates. We used a random effects model to calculate pooled estimates and assessed heterogeneity with I studies,,22 we inIn brief, 21 studies,22–30 inCohort sizes and follow-up times in prospective studies differed considerably, and the number of SAH ranged from 55 to 4373 The repoEight prospective,22,29,30Of the 12 studies,,24,27–30Two population-based studies,,6 includOf the nine,23,25,26Three,5,25 stuNine studies,24,27–30Five,9,11,25 All,24,27–30All nine,23,25,26Five retrospective,5,9–11 aOf the 12,24,27–30Three,6,29 proOne prospective study reported that risk for SAH decreased with an HR of 0.6 per SD unit increase in HDL in the subgroup under age 50 when they adjusted for age, sex, smoking, and alcohol consumption, but not for hypertension .2] Anot Anot2] AOf the 12,24,27–30All nine,23,25,26Since small studies tend to combine SAH and intracranial hemorrhage in a group of hemorrhagic stroke, we excluded them from the analysis.Of the four,6,25,29 2 = 79.5%-89.0%), and considerable in prospective studies (I2 = 46.0%) “Figures A-D in In addition to evident methodological differences among studies “A meta-analysis limited to prospective studies with comparable wide TC groups, major limitations only in an analysis by sex and exclusion of certain SAH types,,24,27,30The meta-analysis including only retrospective studies with TC values measured in an acute phase,11,25 shSome relatively high-quality evidence,6 suggesInternal quality of SAH risk factor studies varies considerably, as most have significant methodological shortages and gaps in reporting. Five studies–6,25 repThe recent low-risk-of-bias study suggested that TC TC and LDL levels are well-documented risk factors for other cardiovascular diseases (CVDs), whereas HDL is generally considered a protective risk indicator. The roleProspective follow-up studies require a significant number approx. 270 000) of person-years and optimally over 35 SAHs for each sex per TC subgroup to reliably study associations between TC levels and risk for SAH. In addition, because roughly 20% of SAH patients die away from hospitals or in emergency rooms, and are 70 000 ofOne of the most important shortcomings in the prospective cohort studies reviewed is their lack of longitudinal follow-up data; changes in risk-factor profile during the study period may therefore distort and confound association analyses. For example, in prospective studies from Norway and FinlThe reliability of retrospective studies in finding new risk factors for SAH is generally limited, not only because all outside hospital deaths are excluded from analyses, but also because of the evident risk of measurement bias when risk factors are measured by interview or after SAH. This was also evident in our sensitivity analyses, which included only on admission TC measurements or measurements by interview. Studies which measured TC by interview suggested that low TC elevates SAH risk. As TC levels may decrease during and after SAH,,46 all sOur review has a few shortcomings. First, we may have missed some relevant publications due to indexing errors and limitations in the search strategy. Our search strategy, however, yielded 21 reviewed studies whereas the three earlier systematic reviews on this topic comprised only 7, 6,13] a a13] andElevated TC levels may elevate risk for SAH among men, and therefore lipid profile should perhaps be taken into account when assessing risk for SAH in men. No convincing evidence exist that low TC elevates risk for SAH. Apart from diabetes and obesity, the risk-factor profile of SAH resembles that of other CVDs, at least in men.S1 Cohorts(DOCX)Click here for additional data file.S1 Figures(DOCX)Click here for additional data file.S1 Methods(DOCX)Click here for additional data file.S1 PRISMA Checklist(DOC)Click here for additional data file.S1 Table(DOCX)Click here for additional data file.


Magnolia officinalis inhibited human UBC cell proliferation, survival, cancer stemness, migration, and invasion, through downregulation of EZH2 expression level, along with the reductions of MMP9, CD44, Sox2 and the induction of tumor suppressor miR-143. Either EZH2 overexpression or miR-143 inhibition could partially reverse honokiol-induced cell growth arrest and impaired clonogenicity. Importantly, it was first revealed that EZH2 could directly bind to the transcriptional regulatory region of miR-143 and repress its expression. Furthermore, honokiol treatment on T24 tumor xenografts confirmed its anticancer effects in vivo, including suppression tumor growth and tumor stemness, accompanied by the dysregulation of EZH2 and miR-143 expressions. Our data suggest a promising therapeutic option to develop drugs targeting EZH2/miR-143 axis, such as honokiol, for bladder cancer treatment.The oncoprotein EZH2, as a histone H3K27 methyltransferase, is frequently overexpressed in various cancer types. However, the mechanisms underlying its role in urinary bladder cancer (UBC) cells have not yet fully understood. Herein, we reported that honokiol, a biologically active biphenolic compound isolated from the  Urinary bladder cancer (UBC) is one of the most common urogenital malignant tumors, with an estimated 74,690 new cases and 15,580 deaths occurring in USA during 2014 , 4.Recent studies revealed that a small population of cancer stem cells (CSCs), which is enriched after therapy, may account for chemotherapy failure -7. TheseHistone modification through polycomb repressive complexes (PRCs) plays an essential role for normal and malignant cell stemness maintenance , 9. As aRecent studies revealed that microRNAs (miRNAs) as a new class of regulators for tumor development. miRNAs belong to the noncoding RNAs with approximately 22 nucleotides, functioning as negative regulators for gene expression at the post-transcriptional level. Previous studies indicated that miR-101 enhanced cytostatic drug sensitivity of liver cancer through targeting EZH2 gene , 15. In Magnolia officinalis. It has been long to be used in traditional Chinese medicine, showing multiple biological activities, including anticancer, anti-inflammation, and neuroprotective properties  = (length [mm]) × (width [mm])2/2. At the endpoint, tumors were weighed and processed for further histological analysis.2.5 × 105 μm-thick paraffin sections were for IHC staining. The primary antibodies against CD44  and Ki67  were incubated overnight at 4°C, respectively. Secondary antibody conjugated with horseradish peroxidase  was then incubated for 1 h at room temperature. The slides were developed with the NovaRED substrate kit  and counterstained by hematoxylin. A score was assigned to represent the estimated percentage of positively stained carcinoma cells as follows: 0: none, 1: ≤50%, 2: 50-75%, and 3: ≥75%. An intensity score was assigned to represent the average estimated intensity of staining in positive carcinoma cells as follows: 0: none, 1: weak, 2: intermediate, 3: strong. The proportion score and intensity score were multiplied to obtain a total score ranging from 0 to 9.t-test .Each experiment was repeated three times. The mean values with standard deviations from the triplicates were plotted. The significant difference between control and experimental groups was analyzed using


The Notch signaling pathway plays a significant role in embryonic cell fate determination and adult tissue homeostasis. Various studies have demonstrated the deep involvement of Notch signaling in the development of the pancreas and the lateral inhibition of Notch signaling in pancreatic progenitor differentiation and maintenance. The targeted inactivation of the Notch pathway components promotes premature differentiation of the endocrine pancreas. However, there is still the contrary opinion that Notch signaling specifies the endocrine lineage. Here, we review the current knowledge of the Notch signaling pathway in pancreatic development and its crosstalk with the Wingless and INT-1 (Wnt) and fibroblast growth factor (FGF) pathways. Hes1 gene by a complex consisting of the Notch intracellular domain (NICD), Rbp-Jκ family of nuclear proteins , Su(H) (Drosophila), and Lag-1 (Caenorhabditis elegans)), and the Mastermind co-activator. However, non-canonical Notch signaling broadly encompasses several modes of Notch activity that do not go through the Rbp-J and activation of the Hes/Hey genes [Notch signaling is an evolutionarily conserved pathway for cell-cell communication and cell-fate determination during embryonic development and tissue homeostasis ,2; it iney genes ,6. Here,ey genes ,8,9. Abney genes ,11,12,13Ngn3), and the formation of insulin-producing β-cells is significantly enhanced by induction of pro-endocrine factors or inhibition of Notch processing. However, recently, some researchers have disagreed with the notion that the Notch pathway is an inhibitor of endocrine cell differentiation. They have proposed that the Notch pathway specifies the pancreatic progenitors differentiating towards endocrine lineage [etc. [Over the past two decades, numerous reports have revealed the pivotal role of Notch signaling in pancreatic specification, cell proliferation, differentiation and plasticity ,14,15,16 lineage ,22 or th lineage ,24,25. S lineage . Studies lineage ,28. Durige [etc. ,30,31. TThe mammalian pancreas is derived from two independent ventral and dorsal buds and experiences three stages of transition ,33. In mPdx1 gene becomes progressively confined to endocrine β-cells, where it plays an essential role in the transcriptional activation of the insulin gene [Ngn3-null mice fail to generate pancreatic endocrine cells and lose the expression of islet transcription factors, including islet-1 (Isl1), NK2 homeobox 2 (Nkx2.2), paired box 4 (Pax4), paired box 6 (Pax6) and NeuroD1, which are all important for endocrine cell differentiation [cyclin D1 in Ngn3+ endocrine progenitors [Ptf1a mutant zebrafish model has found that the down-regulation of Ptf1a induces acinar-to-endocrine fate conversion [et al. [Lineage-specific transcription factors control the differentiation of pancreatic progenitor cells towards a specific type ,41. Panclin gene . The prolin gene ,44  and pyruvate dehydrogenase kinase (Pdk1) genes. miR-19b and miR-18a have been shown to directly act on NeuroD1 and Ptf1a 3′ UTR, respectively [Ngn3 transcript [Growing evidence suggests that miRNAs play an important role in the embryonic development and physiological function of pancreas. Examples of these miRNAs include miR-7, miR-375, the miR17-92 cluster, miR-26 and miR-15 et al. . Dicer-nget Hes1 . miR-7 iget Hes1 . miR-375get Hes1 . We haves1 [Yap1 . miR-375ectively ,61. Thesanscript .The development of the pancreas depends on the spatio-temporal expression of a number of transcription factors and associated signaling pathways ,64. StudHes) and Hairy/enhancer of spit related with YRPW motif (Hey) families, nuclear factor-κB (NF-κB), vascular growth factor receptor (VEGF), etc. [Hes genes, which encode bHLH transcriptional repressors, play crucial roles in the fate choice and differentiation of stem cells, e.g., in the inner ear, neurons and pancreas development [Ngn3 by binding to the proximal promoter and specifically blocking promoter activity [Notch genes encode a single-pass transmembrane receptor family of molecules, including Notch1–4 in mammals . Notch pF), etc. ,71  competes with Maml to repress the Notch1 target genes [+ cells, and the ectopic expression of Mfng is sufficient to induce chick endodermal cells to differentiate towards endocrine fate [Mfng in mice has no effect on pancreatic development, cell differentiation or function [Several Notch-related regulators have also been found to function in normal organism development and tumorigenesis. In the left-right asymmetric development of et genes . Mind boet genes . It is ret genes ,80. In net genes . DEAD  [Notch1 can block the expression of endocrine genes and prevent endocrine differentiation [Hes1 and intracellular domain of Notch (Notch-IC) in mouse dorsal pancreatic buds at E10.5 represses endocrine and exocrine differentiation [Notch activation in zebrafish embryos can inhibit acinar and β-cell differentiation [Notch was overexpressed in the E9.5 or E11.5 mouse embryonic pancreas, the differentiation of Hes1+ cells was blocked; however, ectopic Notch activation at E15.5 does not perturb exocrine differentiation [During the secondary transition, Notch regulates endocrine differentiation via a lateral inhibition mechanism. The lateral inhibition model proposes that the onset of Ngn3 expression initiates endocrine differentiation and activates the Notch ligand Delta. Subsequently, Delta binds with Notch receptors in neighboring cells to initiate the Notch signaling cascade and release the NICD. The activated NICD enters the nucleus to activate the target gene  of Ngn3 ,76. Ulti of Ngn3 . In Dll1ne cells . In zebrntiation . The los3+ cells . Furtherdiabetes ,102. Hesne cells ,103. A sls [Ngn3  or the ignaling) . Conversntiation . The lenntiation . Ectopicntiation . When Nontiation . The susntiation .Dll1 mutant mice show a dramatic decrease in their number of Ngn3+ cells, which suggests that Dll1 is required for the continuous formation of Ngn3+ endocrine precursors [et al. [Ngn3 gene activator Sox9, which may provide further evidence that Notch initiates the endocrine lineage. Some current gene-targeting studies focused on the Notch signaling components found that inactivated Notch signaling promotes the differentiation or transdifferentiation of MPCs, CACs or adult duct cells to the acinar lineage, which suggests that Notch may specify the endocrine lineage. Zebrafish embryos that were injected with RNA encoding a dominant-negative Suppressor of Hairless showed accelerated exocrine cells compared with the controls [Psen1) and presenilin2 (Psen2) drives Ngn3-expressing cells to differentiate into acinar cells [Mib1 in the endoderm causes a loss of endocrine cells and an increase in exocrine cells in the proximal domain, which suggested that Notch signalling is required to prompt MPCs to adopt an endocrine fate [Rbp-J deletion, mouse CACs undergo a rapid conversion to acinar cells [Hes1 depletion but is suppressed by NICD induction in adult mouse Sox9-expressing cells [Furthermore, recent studies have implied that the Notch-mediated regulation of endocrine differentiation is more complex than the classical lateral inhibition model suggests. For example, E9.5 ecursors . Shih et [et al. , demonstcontrols . Similarar cells . Once cuar cells . When Noar cells . The inaine fate . Upon Rbar cells . In adding cells .et al. [et al. [The causal role of Notch signaling in the patterning of MPCs into endocrine cells has been evaluated in some studies in different aspects. Genetic interactions have suggested that γ-secretase and Notch2 act in a non-canonical mechanism to sequester Rbp-J away from Ptf1a, which secures mouse Ngn3-positive progenitor cells to the endocrine fate . Afelik et al. , found tet al. ,26,76. A [et al. , proposeThe Notch signaling pathway has been found to maintain proliferation and prevent the precocious differentiation of pancreatic progenitor cells ,110. TheP57 gene, which leads to cell cycle arrest, early differentiation and the depletion of the progenitor pool [Sox9-deficient progenitors exhibit reduced proliferation and a low level of Hes1 [Sox9 resulted in a decrease in pGSK3β, nuclear β-catenin and the Wnt signaling target gene cyclin D1 [CreERT2Sox9 mice [The transcription factor network is crucial for the maintenance and expansion of MPCs. Pdx1 and Ptf1a can form autoregulatory loops and feed-forward loop to retain and expand the progenitor cells during early pancreatic development . The inator pool . Moreovetor pool ,45,118.  of Hes1 . In isolyclin D1 ,25. NotcHes1, Hey1, and Hey2 [in vitro [et al. [Because the components of the Notch signaling pathway are not normally expressed in terminally differentiated pancreatic cells, most mature cells lose their responsiveness to Notch signaling. However, in-depth studies have indicated that Notch signaling is involved in controlling the plasticity of adult, terminally differentiated pancreatic cells. Importantly, the dedifferentiation and transdifferentiation of terminally differentiated pancreatic cells are associated with Notch pathway reactivation during regeneration following pancreatitis ,99,119, and Hey2 . Adult pand Hey2 . When diand Hey2 . When Heand Hey2 . Notch sin vitro ,125. Bae [et al. , found tWnt7b leads to pancreatic hypoplasia because of the reduced proliferation of pancreatic progenitor cells [β-catenin before E11.5 in mouse embryos prevents the proper differentiation and expansion of early pancreatic progenitor cells [β-catenin in mouse embryos at E18.5 causes the gross enlargement of the exocrine pancreas, which results in a dramatic increase in pancreas organ size [β-catenin knockout mice, the pancreas almost completely lacks acinar cells [Notch and Wnt signaling are key pathways that control the expansion and differentiation of stem/progenitor cells during embryogenesis, tissue formation and maintenance in adult homeostasis . The Wntor cells . The ector cells . Ectopicgan size . Conversar cells . Meanwhiar cells ,132. Conar cells . Howeverar cells ,131. Likar cells ,134. Thear cells .Drosophila and Xenopus [Drosophila [et al. [Xenopus epidermal development , which regulates pancreatic epithelial growth and differentiation by suppressing Notch signaling in mice [FGF10 is similar to that of the mice with Notch overexpression described above. Thus, Notch signaling is a critical downstream effector of FGF pathway-induced embryonic pancreatic epithelial proliferation . Initially, the Notch pathway enhances the neural precursor proliferation and represses their differentiation ,161. At


Paenibacillus strain P1XP2, identified genes for enzymes with the potential for degrading complex food wastes, a property consistent with the ecological habitat of the isolate.The analysis of the ~5.8-Mb draft genome sequence of a moderately thermophilic, heterotrophic, facultative anaerobic bacterium,  Paenibacillus strain P1XP2  is a heterotrophic, moderately thermophilic , facultative anaerobic, and fermentative bacterium isolated on 0.1% tryptic soy broth (TSB) from a sample collected from Baku Baku, a commercial bioreactor degrading food waste  to mid-late log growth phase, the cells were centrifuged, and the DNA from the cell pellet was purified using a modification of Marmur’s method . A large number of genes involved in carbohydrate degradation (639), including genes for chitinase, α-amylase, pullulanase, cyclodextrinase, glucosidase, galactosidase, glucuroxylanase, and arabinogalactan endo-1,4-beta-galactosidase, together with a slightly smaller number of genes (339) involved in protein and amino acid degradation and synthesis, were identified in the genome following RAST analysis. The presence of xylanase and amylase in strain P1XP2 was supported by activity screening using X-xyl and starch agar plate assays  has the accession number JRNV01000000 and consists of sequences JRNV01000001 through JRNV01000186. The version described in this paper is version JHCA00000000.1.The


Each cell of the immune system shows a characteristic expression of cell surface molecules depending on the grade of activation or suppression. We examined the cell surface profile on polymorphonuclear cells (PMN) of polytraumatized patients and compared them with those of healthy controls and furthermore with patients with severe sepsis or septic shock. Moreover, we wanted to find out in the polytraumatized patients whether there are associations of PMN surface marker patterns with severity of injury, of disease and of organ dysfunctions.In a prospective observational study, cell surface expression of CD16, CD88, CD64, CD66b, CD11b, CD95, CD33, CD39, IL-17RA, TLR- 2, TLR-4 and HLA-DR on PMN was determined using flow cytometry. Thirteen healthy volunteers, eight patients with severe sepsis and septic shock, and longitudinally, on admission (0 hours), and 4, 12, 24, 48, 120 and 240 hours thereafter, eight polytraumatized patients were monitored. ISS, SAPS3 and SOFA score reflect severity of injury, of disease and of organ dysfunctions.+ expression and CD33+/CD66b− expression were significantly lower and CD33/CD66b+ higher than in healthy controls. Especially, the lowest values of CD88+ occurred in patients with polytrauma developing septic complications.In polytraumatized patients (24 hours measurement), but also in septic patients, CD88+ was lower. Considering the post-traumatic course, polytraumatized patients with better prognostic evaluation in SOFA andSAPS3 initially expressed higher levels of TLR-2. Polytrauma patients could be divided into two subgroups, one with and the other without septic complications. The septic subgroup presented predominantly higher values in SOFA, SAPS3 and ISS scoring.Furthermore, the polytraumatized patients revealed higher levels of CD66b, CD11b and CD16, whereas CD16Cell surface molecules on PMN of polytraumatized patients differ from those of healthy volunteers. Besides, distinct expression patterns resemble those of patients with sepsis. Patents with higher ISS values go along with higher scores in SOFA and SAPS3, and are at risk to develop septic complications.


Acinetobacter baumannii has emerged as a dangerous nosocomial pathogen, particularly for severely ill patients in intensive care units and patients with hematologic malignancies. Here, we present the complete genome sequence of a multidrug-resistant A. baumannii isolate, recovered from a Mexican hospital and classified as sequence type 422 according to the multilocus sequence typing Pasteur scheme. Acinetobacter baumannii is a Gram-negative bacterium that has emerged as a dangerous pathogen worldwide, causing a variety of nosocomial infections, especially in severely ill patients in intensive care units and patients with hematologic malignancies on chemotherapy  was constructed and sequenced, using one SMRT cell with a P6 polymerase and C4 chemistry combination (P6-C4) with a 180-min movie. The SMRT cells produced 551,632,314 post-filter polymerase reads with a mean read length of 10,718 bp. Subreads were assembled de novo using the PacBio RS hierarchical genome assembly process (HGAP) protocol version 3 in SMRT analysis version 2.3 (Pacific Biosciences)  and two plasmids: pAba3207a  and pAba3207b . The chromosome has six rRNA operons, 74 tRNAs, and 3,674 coding sequences (CDSs) and embraces most of the antibiotic resistance genes, conferring resistance to aminoglycosides, beta-lactams, carbapenems (blaOXA-65), cephalosporins, fosmidomycin, fusaric acid, phenicols, and tetracyclines. The smallest plasmid has 19 CDSs, including one carbapenem resistance gene (blaOXA-58), whereas pAba3207b has 97 CDSs.The A. baumannii in Mexico, not only at some loci but at the genome level. However, to get a more comprehensive picture, comparative analyses of many more isolates are warranted.This and a previous report  are the CP015364, CP015365, and CP015366.The genome sequence of isolate 3207 was deposited in GenBank under the accession numbers


Genome replication in flavivirus requires (−) strand RNA synthesis, (+) strand RNA synthesis, and 5′-RNA capping and methylation. To carry out viral genome replication, flavivirus assembles a replication complex, consisting of both viral and host proteins, on the cytoplasmic side of the endoplasmic reticulum (ER) membrane. Two major components of the replication complex are the viral non-structural (NS) proteins NS3 and NS5. Together they possess all the enzymatic activities required for genome replication, yet how these activities are coordinated during genome replication is not clear. We provide an overview of the flaviviral genome replication process, the membrane-bound replication complex, and recent crystal structures of full-length NS5. We propose a model of how NS3 and NS5 coordinate their activities in the individual steps of (−) RNA synthesis, (+) RNA synthesis, and 5′-RNA capping and methylation. Flaviviridae, which also includes Hepacivirus and Pestivirus. The Flavivirus genus includes over 70 viruses, many of which cause arboviral diseases in humans, such as dengue (DENV), West Nile (WNV), tick-borne encephalitis (TBEV), and yellow fever virus. The 11 kb flaviviral RNA genome consists of a 5′-cap, a 5′-untranslated region (5′-UTR), a single open reading frame (ORF), and a 3′-UTR. The 5′- and 3′-UTRs contain conserved RNA secondary structures that are important for viral replication, including sequences that mediate long range 5′- and 3′-RNA interactions [Flaviviruses are positive (+) sense RNA viruses belonging to the family ractions ,5,6,7,8.ractions . Among Nractions ,13,14. Tractions ,17,18,19ractions ,21. NS5,ractions . The NS5ractions ,24. Littractions ,27,28,29ractions ,31,32,33ractions ,35. Moreractions ,37.m7GpppNm-RNA) at its 5′ end. RNA synthesis in flavivirus is semi-conservative and asymmetric. The (+) sense RNA, which is the same polarity as the viral genome, is predominantly formed over the (−) sense RNA; single-stranded (ss) RNA found in flavivirus-infected cells is (+) sense RNA, and (−) strand RNA is only detected in the dsRNA form [3H-UTP or 32P-labeled guanosine-5'-triphosphate (GTP)), three radiolabeled RNA products were identified: a double-stranded replicative form (RF) RNA, single-stranded genome length RNA, and the slowest migrating, replicative intermediate (RI) [The flavivirus replication complex carries out RNA synthesis, RNA capping and RNA methylation steps to produce the genome with a type 1 cap structure (RNA form . When DEate (RI) ,39,40. Rate (RI) ,39. BaseThe (+) strand progeny RNA is subsequently capped at its 5′ end and methylated to form a type 1 cap B. The caGenome replication in flavivirus is carried out by a membrane-bound viral replication complex consisting of viral NS proteins, viral RNA and unidentified host proteins . Membranin vitro flaviviral replication system, which can use the viral genome to synthesize the methylated, capped RNA product, is currently unavailable.Although the exact composition of the replication complex is not known, all flaviviral NS proteins have been shown to be a component of the replication complex. Using immunoprecipitation, yeast two-hybrid, and fluorescence resonance energy transfer (FRET) assays, interactions among NS proteins have been identified ,46,47,48in vivo fluorescent measurements, and yeast two-hybrid studies [At the heart of the flaviviral replication complex are NS3 and NS5, which together account for all enzymatic activities required to amplify the RNA genome and to attach a type 1 cap to its 5′ end . Other v studies ,47,57. T studies ,59,60.The physical linkage between the MTase and RdRp domains within NS5 suggests that viral genome replication and 5′-RNA capping may be coordinated between the two domains. Several genetic studies show that mutations in the MTase domain impact RNA replication by the RdRp domain, suggesting that the RdRp and MTase interact during viral genome replication ,62,63,64260Gly-Thr-Arg262 (GTR) motif, N-terminal to the inter-domain linker, is proposed to act as a hinge that allows movement of the MTase and RdRp domains relative to one another [The recently reported structures of full-length NS5 from JEV and DENV offer details of specific interactions between the MTase and RdRp domains and suggest a structural mechanism by which their activities may be coordinated within the flavivirus replication complex ,69. We h another ,69,70. AThe multifunctional flavivirus NS5 carries out several reactions. These include synthesis of (−) strand RNA, synthesis of (+) strand RNA, addition of a guanine cap to the 5′ end of (+) strand RNA, and two methylation reactions to form a type 1 cap structure at the 5′ end of the nascent RNA. Consequently, NS5 interacts with other viral proteins and several different forms of viral RNA, including both single- and double-stranded, (+) and (−) strand, capped and uncapped, and methylated and unmethylated forms of capped RNA. Thus, NS5 likely needs to adopt multiple conformations during different stages of genome replication and processing, as suggested by the conformational variation observed for the structures of full-length NS5 from JEV and DENV. Below, we summarize viral replication steps in terms of coordination of NS3 and NS5 functions.(−) strand RNA synthesis: (−) sense RNA synthesis is carried out by NS5 polymerase using the viral (+) RNA genome as a template. The function of NS3 does not seem to be required for (−) strand RNA synthesis, since NS5 (either the full-length protein or the polymerase domain) alone is capable of synthesizing RNA using viral subgenomic RNA. The (+) sense RNA genome contains cyclization sequences at the 5′- and 3′-UTR, and the cyclized genome serves as the template for (−) strand RNA synthesis ,7,8,71. (+) strand RNA synthesis: The viral replication complex uses the (−) strand RNA in the dsRNA intermediate as the template to synthesize multiple copies of (+) sense RNA. Both NS5 and NS3 activities are required for (+) RNA synthesis . NS3 hel5′-RNA capping and methylation of (+) sense RNA: The (+) sense RNA will be capped and methylated to form a type 1 cap at the 5′ end B. NS3 heCoordination of (+) RNA synthesis and 5′-RNA capping: The 5′-RNA capping and methylation steps are likely coupled with (+) sense RNA synthesis, but during which step of RNA synthesis 5′-RNA capping and methylation occur is not clear. Capping of 5′-RNA could occur on the nascent (+) strand RNA while it is being synthesized by the NS5 RdRp domain (co-transcriptional model in The identification and visualization of subcellular membrane structures used by flavivirus as replication factories have provided insight into how flavivirus may orchestrate viral replication and virion assembly in infected cells. Additionally, crystal structures of the full-length NS5 have provided details of intra- and inter-molecular interactions that stabilize different forms of NS5. These forms may interact with RNA and protein components of the replication complex differently and thus serve as a platform to promote different steps during genome replication. Future studies will be geared toward understanding how these membrane vesicles are related to the assembly and function of the viral replication complex at the molecular level. In particular, how flaviviral and host proteins establish such elaborate membrane vesicles for replication, how flaviviral NS proteins are assembled in the membrane-bound viral replication complex, and how the viral replication complex carries out individual steps of RNA replication are of great interest.


Aspergillus, Penicillium, Cladosporium, and Fusarium were subjected to compound isolation and antiviral studies, which led to an illustration of the strong antiviral activity of a variety of marine fungi-derived compounds. The present review strives to summarize all available knowledge on active compounds isolated from marine fungi with antiviral activity.Biodiversity of the marine world is only partially subjected to detailed scientific scrutiny in comparison to terrestrial life. Life in the marine world depends heavily on marine fungi scavenging the oceans of lifeless plants and animals and entering them into the nutrient cycle by. Approximately 150 to 200 new compounds, including alkaloids, sesquiterpenes, polyketides, and aromatic compounds, are identified from marine fungi annually. In recent years, numerous investigations demonstrated the tremendous potential of marine fungi as a promising source to develop new antivirals against different important viruses, including herpes simplex viruses, the human immunodeficiency virus, and the influenza virus. Various genera of marine fungi such as  The marine world contains approximately one half of all species ,2. The vAspergilus and Fusarium solani contributed to the mortality of the Caribbean sea-fan and the infections of different marine crustaceans, respectively [Lagenidium callinectes [Historically, the pivotal role of fungi in different aspects of human life is very pronounced and this is true even in the marine world . Marine ectively ,15. In alinectes .Turgidosculum ulvae can only be grown in the thallus of Blidingia minima, a green algae [Penicillium and Aspergillus species in the marine environment are isolated from sponges [Despite the pathogenicity of certain marine fungi species, mutualistic interactions are the dominant types of relationship found in marine fungi . The lifen algae . Moreove sponges . Isolati sponges .As scientific interest has been sparked in marine microorganisms, fungi and their metabolites have begun to be recognized for their potent biological activities in the past few decades. Some of these metabolites give marine fungi the superiority to adapt to extreme habitats, compete for substrates, and ward off threats . Moreoveetc. or mimicking natural products were much higher. This highlights the potential role of natural products in the further establishment of new antiviral agents [Stachybotrys sp. RF-7260 by Taishi and colleagues; it had promising antiviral activity against influenza A virus (H1N1) [Viruses encompass a prodigious group of microorganisms causing assorted infectious diseases. Recent scientific studies triumphantly reported new antiviral agents, which generally inhibit the virus replication cycle through affecting the important host cell factor(s) for virus replication and/or viral elements . Despitel agents . The focs (H1N1) . Until 2s (H1N1) . HoweverPicornaviridae family, has emerged as a critical non-polio neurotropic enterovirus. EV71 provokes acute neurological disease in children, which may result in cardiopulmonary failure and death. However, this non-enveloped, positive (+) strand RNA virus is a major cause of hand, foot, and mouth disease [After the virtual eradication of poliovirus, enterovirus 71 (EV-71), a member of the  disease . During  disease . Despite disease .1), grisephenone A (2), and 3,6,8-Trihydroxy-1-methylxanthone (3), which are new sesquiterpenoid and xanthone derivatives isolated from the cultures of sponge-derived fungus Stachybotry sp. HH1 ZDDS1F1-2, have been evaluated against in vitro replication of EV-71. The so-called compounds showed inhibitory activities against in vitro replication of EV71 with IC50 values of 30.1, 50.0 and 40.3 μM [The antiviral activity of three compounds, stachybogrisephenone B —namely, genital herpes—that also can be transmitted to neonates during delivery through the infected mother’s birth canal. Despite numerous reports of natural products with inhibitory effects against HSV, commercial drugs are generally limited to synthetic antiviral agents with a natural product pharmacophore such as acyclovir, penciclovir, and famciclovir [HSV-1 and HSV-2 are important and prevalent human pathogens from ciclovir ,59.4–8), a series of lipophilic linear peptides that were isolated from the genus Scytalidium during saline fermentation [in vitro antiviral activity against HSV-1 and HSV-2 in a time- and dose-dependent manner. This activity was suggested to be through viral membrane destabilization. These fungal metabolites exert direct virucidal activity against extracellular HSV particles as an applicable approach to the inhibition of HSV transmission [Halovirs A–E , arisugacin A (10), isobutyrolactone II (11), and aspernolide A (12), with IC50 values against HSV-1of 33.38, 12.76, 62.08, and 68.16 μM, respectively. These compounds derived from a marine fungus, Aspergillus terreus SCSGAF0162, which was isolated from the South China Sea  gorgonian corals Echinogorgia aurantiaca, under solid-state fermentation of rice.Nong and colleagues (2014)  reported13), is one of the antiviral naphthalenone byproducts from the ethyl acetate (EtOAc) extract of the culture broth of fungal strain 222 belonging to the Ascomycota, which was obtained on driftwood gathered from the Greifswalder Bodden shore, Baltic Sea, Germany. To evaluate the antiviral activity of balticolid against influenza A virus and HSV-I, the non-cytotoxic concentrations of the compound were examined on in vitro replication of both viruses. Balticolid showed potent inhibitory effects against HSV-I with IC50 = 0.45 μM. However, there was no significant antiviral activity against in vitro replication of the influenza A virus [A new 12-membered macrolide, balticolid  [HIV causing acquired immunodeficiency syndrome (AIDS) is the major contributing factor for the elevation of infectious disease mortality all around the world . This me agents) . However agents) ,63.14) and its new enantiomeric homologue, phomasetin (15), isolated from the marine fungi Fusarium heterosporum and Phoma sp., respectively, exhibited in vitro inhibitory activity against the integrase enzyme of HIV-1, which is an important enzyme for the HIV-1 replication cycle. Further characterization of the active compound from a marine fungal species, Xylaria sp., yielded integric acid (16), which structurally is similar to equisetin (14) as a novel and unique class of integrase inhibitors [impeded 3́end activity and strand transferal reactions, both with IC50 = 10 μM.Equisetin  strand RNA virus and has several subtypes named based on the H (hemagglutinin) and N (neuraminidase) antigens. Birds are the main host for influenza A virus; however, transmission to domestic poultry may infect some mammals and provoke dangerous human influenza pandemics [Every few years, emergence of a novel influenza virus with fatal pathogenicity attracts global attention to the marked numbers of afflicted patients and the high rate of death; still human beings are confounded by virus recombination . Reformuandemics . Respiraandemics .17) was isolated from Stachybotrys sp. RF-7260 by the solid-state fermentation method. This terpenoid elicited an IC50 = 0.003 μM against influenza A virus (H1N1), which was comparable with other antiviral agents such as zanamivir and amantadine [in vivo study by oral administration of stachyflin and its derivatives with a solution in PEG [Stachyflin , with potential antiviral property against H1N1 strain of influenza virus. The resultant compounds, oxoglyantrypine (18), norquinadoline A (19), deoxynortryptoquivaline (20), deoxytryptoquivaline (21), tryptoquivaline (22), and quinadoline B (23) revealed noteworthy antiviral activities against H1N1 strain with IC50 values of 85, 82, 87, 85, 89, and 82 μM, respectively [Cladosporium sphaerospermum 2005-01-E3, obtained from residues in the Pacific Ocean. The novel compound cladosin C (24) showed moderate antiviral activity against influenza A (H1N1) virus with an IC50 = 276 μM.In light of investigations for discovering anti-influenza compounds, an effective mangrove-derived fungal strain, ectively . In a laectively  evaluateAspergillus sydowii, which was isolated from an unknown marine sponge gathered from the Xisha Islands of China. The derived compounds, (Z)-5-(Hydroxymenthyl)-2-(6′)-methylhept-2′-en-2′-yl)-phenol (25), diorcinol (26) and cordyol C (27), exhibited minor antiviral activity against the influenza (H3N2) virus with IC50 values of 57.4, 66.5, and 78.5 μM, respectively [Aspergillus terreus (OUCMDZ-1925 strain) by Zhu and colleagues [28) presented prominent antiviral activity against influenza A (H1N1) virus with an IC50 = 87.1 μM.In addition, a recent study identified new bioactive natural compounds from a sponge-associated fungi, the ZSDS1-F6 strain of ectively . Furtherlleagues  ,of whic29), a cyclic tetrapeptide compound with antiviral properties that derived from the fungal strain of A. terreus SCSGAF0162, was isolated from the tissue of the gorgonian Echinogorgia aurantiaca, collected from Sanya, Hainan Province, China. 29 displayed suppressive impacts on the influenza virus strains A/WSN/33 (H1N1) and A/Hong Kong/8/68 (H3N2) with IC50 values of 15 and 8.1 μM, respectively [A.terreus Gwq-48, which was derived from a mangrove rhizosphere soil sample in the coast of Fujian state. The analysis of its chemical ingredients resulted in the segregation of one novel aspulvinone, isoaspulvinone E (30), aspulvinone E (31), and pulvic acid (32), which exhibited notable anti-influenza A (H1N1) virus activities, with IC50 values of 101.23, 192.05, and 94.42 μM, respectively [Asperterrestide A (ectively . FurtherEmericella sp. (HK-ZJ), isolated from the internal rind of a mangrove plant Aegiceras corniculatum in the district of HaiKou, China [33 & 34), emeriphenolicins A and D, aspernidine A and B, austin, austinol, dehydroaustin, and acetoxydehydroaustin, for their in vitro effect against replication of H1N1 in MDCK cells, just two compounds, 33 and 34, displayed average suppressive effects with IC50 values of 201.1 and 296.62 μM, respectively.An investigation with the goal of producing an anti-influenza virus resulted in the isolation of a fungus verified as u, China . FollowiPenicillium purpurogenum, was derived from the regional red soil by the Lujiang River from Jianshui, Yunnan, China. Subsequently, the antiviral activity was detected for compounds purpurquinones B and C (35 & 36), purpuresters A (37), and TAN-931 (38) against H1N1, with IC50 values of 61.3, 64.0, 85.3, and 58.6 μM, respectively [Penicilliumchrysogenum PJX-17, two novel compounds, sorbicatechols A and B (39 & 40), were properly isolated from a sorbicillin component and a styrene moiety by endo- and exo-Diels—Alder cycloadditions, and both of them showed anti-H1N1 activity, with IC50 values of 85 and 113 μM, respectively.A recent study on novel drugs against the influenza virus from extremophiles, an acid-tolerant fungal strain JS03-21, determined as ectively , on the Arteriviridae family with a complex epidemiological profile due to the persistence of the virus in tissue even several months after the acute stage. This small, enveloped, positive (+) strand RNA virus infects pigs and causes respiratory illness and a major failure in the reproduction of sows. An RNA genome of about 15 kilobases in size is responsible for the coding of two principal polyproteins, namely 1a and 1ab [PRRSV is a member of the  and 1ab . PRRSV c and 1ab . Assorte and 1ab . Despite and 1ab .41) and alterporriol Q (42) were isolated from the marine-derived fungus ZJ-2008003, obtained from Sarcophyton sp., a soft coral in the South China Sea. Molecular and morphological characterizations identified the isolated species as an Alternaria sp. The compounds 41 and 42 revealed antiviral activities against PRRSV with IC50 values of 65 and 39 μM, respectively [A recent investigation by Zheng and colleagues  reportedectively .Poxviridae, MCV has a unique mode of replication in the human epidermis: itprecipitates a common skin infection, especially in children, through augmentation of epidermal cell mitosis and disruption of cell differentiation. However, similar to other species in this family, this linear, double-stranded DNA virus is characterized by lateral bodies, a core, an envelope, and a surface membrane. The viral genome and replication afflicts several cellular pathways, including cell death, cell cycle, inflammation, and innate immunity. To escape from immune surveillance, the MCV virus does not pass the basement membrane of the epidermis. MCV skin diseases can be comfortably recognized, and a combination of topical treatment and local physical therapy can efficaciously obliterate the virus infection [As a member of the nfection .43), isolated from a marine fungus, Fusarium sp. [50 = 124 μM) was induced through suppression of topoisomerase-mediated DNA-binding, DNA relaxation, and formation of covalent complex [Sansalvamide A -21-epi-aflaquinolone B (44) exhibited exceptional anti-RSV activity with an IC50 value of 42 nM, roughly 500 times more potent than ribavirin, the positive control used in this study, with IC50 = 20 μM [50 = 6.6 μM).Screening of an extract isolated from  = 20 μM . In addiAs a plant virus, TMV markedly debilitates the horticulture and agriculture industries . This ro1 from Penicillium oxalicum exhibited a strong inhibitory activity against the replication of TMV [H)-one (45) and methyl 4-hydroxyphenylacetate (47) possessed strong suppressive activity against TMV with EC50 values of 399.57 μM and 829.15 μM, respectively; while, compared to those, the new compound 2-(4-hydroxybenzoyl) quinazolin-4(3H)-one (46) revealed average suppressive activity against TMV (EC50 was not tested) [Neosartorya fischeri strain 1008F1. The bioactive assays signifying those two compounds, AGI-B4 (48) and 3,4-dihydroxybenzoic acid (49), showed strong inhibitory impact on the replication of tobacco mosaic virus (TMV), with IC50 values of 260 and 630 μM, respectively.Recent surveys reported that the extract of strain 0312Fn of TMV ,56. Bioa tested) . Prior tin vitro studies [in vivo and even pharmacological studies for some of the abovementioned effective compounds seems to be crucial.The increasing rate of viral resistance to antiviral drugs and drug toxicity is becoming a challenging problem in antiviral therapy. There are numerous reports on the resistance of different viruses to approved antiviral drugs ,82,83. H studies ,84,85. TAspergillus sp., Penicillium sp., Cladosporium sp., Stachybotrys sp., and Neosartorya sp.; they are summarized in 13, a newly derived strain of Ascomycete, revealed marked inhibitory activity against HSV. Furthermore, 17 prompted a potent anti-influenza virus activity by showing a very low IC50 value (0.003 µM) and also oral administration of 17 in an in vivo study in PEG confirmed its significant antiviral activity. Furthermore, 44 exhibited a notable potency to elicit substantial antiviral activity against RSV. Nonetheless, the majority of the investigations were limited to basic screening and no mechanism of action was established for active compounds. It is pivotal for further research to characterize and determine the virus or host factors, which were targeted by antiviral compounds.Organisms inhabiting the marine environment provide a diversity of bioactive compounds, which are exclusive as the aqueous habitat demands molecules with particular and vigorous biological compounds. Many scholars are devoted to investigating marine organisms to determine the development of potential biomolecules into therapeutic drugs and numerous compounds from marine fungi have been shown to possess notable antiviral activities. On the other hand, there are many important animal and human viruses yet to be studied, since for most viral diseases there has not been any effective therapeutic treatment available thus far. Therefore, infectious viruses with widespread prevalence, including EV71, HSV, HIV, MCV, and RSV, were used to examine the antiviral potential of the isolated compounds. Moreover, viruses responsible for important plant and animal infections, namely TMV and PRRS, were also employed in several studies. All together, the results showed quite noticeable cytotoxic effects against the respective viruses. From the preceding statement, we presented various compounds isolated from different marine fungi genera of which the most important ones exploited for their antiviral potential were in vivo and clinical studies are other aspects that should be exploited. The variety of the natural products from marine fungi evidently determines the potential for assigning some selected compounds to in vivo and probably clinical trials for forthcoming progress of anti-infective drugs. One of the considerable upcoming challenges will be the extensive production of these compounds to meet the demand for clinical trials and drug development. Several investigators believe that a specific form of combined genetic and metabolic engineering will be the potential resolution for commercial manufacture of these compounds [To develop antiviral drugs derived from marine fungi, ompounds . It is h


Parkinson's disease (PD)—classically characterized by severe loss of dopaminergic neurons in the substantia nigra pars compacta—has a caudal-rostral progression, beginning in the dorsal motor vagal nucleus and, in a less extent, in the olfactory system, progressing to the midbrain and eventually to the basal forebrain and the neocortex. About 90% of the cases are idiopathic. To study the molecular mechanisms involved in idiopathic PD we conducted a comparative study of transcriptional interaction networks in the dorsal motor vagal nucleus (VA), locus coeruleus (LC), and substantia nigra (SN) of idiopathic PD in Braak stages 4-5 (PD) and disease-free controls (CT) using postmortem samples. Gene coexpression networks (GCNs) for each brain region (patients and controls) were obtained to identify highly connected relevant genes (hubs) and densely interconnected gene sets (modules). GCN analyses showed differences in topology and module composition between CT and PD networks for each anatomic region. In CT networks, VA, LC, and SN hub modules are predominantly associated with neuroprotection and homeostasis in the ageing brain, whereas in the patient's group, for the three brain regions, hub modules are mostly related to stress response and neuron survival/degeneration mechanisms. GloThis scenario started to change with the introduction of statistical and computational tools for analyzing gene-gene interaction networks and the comparative analysis of gene expression and interactome . EdwardsThe relevant genes to the pathogenesis of complex diseases, like idiopathic PD, are those with a high number of gene-gene links in transcriptional interaction networks, which do not necessary show what a high differential expression , 16. The3) were homogenized with Tissue Rupter , and total RNA was extracted from the homogenates using the RNeasy Lipid Tissue Kit . RNA quality was assessed on the Agilent BioAnalyzer 2100 . RNA integrity number (RIN) values were all within the acceptable range (6-7) for microarray assays using brain bank samples [P < 0.005 or P < 0.01, resp.) was used. All microarray data were deposited in GEO public database (http://www.ncbi.nlm.nih.gov/geo) under accession number GE43490. Transcriptional interaction network for differentially expressed GO annotated genes was constructed based on Pearson's correlation, using software R. Data analysis and visualization were achieved through Cytoscape software 2.8.0 (http://www.cytoscape.org/).Postmortem samples of dorsal motor vagal nucleus (VA), locus coeruleus (LC), and substantia nigra (SN) from controls and PD subjects  were obt samples . To dete samples  we selecThe comparative analysis of VA, LC, and SN transcriptomic profiles for patient (PD) versus control (CT) groups revealed 234, 183, and 326 differentially expressed GO annotated genes, respectively. All genes were upregulated in PD groups. Transcriptional interaction networks for each anatomic region  for both groups were obtained and analyzed. In each of these six networks, the genes with the higher number of gene-gene links were considered hubs , 23 and k for nondirected networks was calculated by k = 2L/N, where L stands for the number of edges and N for the number of nodes [k values were 6.5 for VA-CT and 7.25 for VA-PD. A total of 178 genes and 646 gene-gene links (threshold 0.94) for VA-PD network and 206 genes and 670 gene-gene links (threshold 0.95) for VA-CT network were obtained, respectively . CPNE2 and HRC are genes involved in brain Ca2+ metabolism and functions: HRC, or histidine-rich calcium binding protein, regulates Ca2+ homeostasis [ CPNE2 acts as a Ca2+ sensor in postsynaptic events [ SOX10 and ZEB2 play a role in myelination processes: SOX10 codes for a transcription factor acting in regulating myelination in oligodendrocytes [ ZEB2, which codes for a Smad-interacting protein, acts in myelination of the central nervous system [The first clusters wereeostasis , whereasc events . SOX10 adrocytes , and ZEBs system  and regus system . S100A4, PGM3, and FLYWCH1. S100A4 codes for a Ca2+-binding protein were involved in neuroprotection, rescuing neurons via the Janus kinase/STAT pathway and, partially, via interleukin-10 receptor [ FLYWCH1 codes a FLYWCH-type zinc finger 1 chromatin modulator protein. Cellular proteins that harbor the FLYWCH domain are predominantly involved in transcriptional regulation [2+ homeostasis, myelination, and neuron differentiation.The second module harbored the genesreceptor , and proreceptor . PGM3 coreceptor  and causreceptor . FLYWCH1gulation . Altoget ADAM15, which codes for a disintegrin metalloprotease, has been implicated in both the process of neuronal hypoxic injury [ SHARPIN codes for a ubiquitin-binding and ubiquitin-like-domain-containing protein, which is an important component of the linear ubiquitin chain assembly complex (LUBAC) that modulates activation of NF-κB signaling pathway, thus controlling cell survival and apoptosis [ GNL3 codes for a nucleolar protein which stabilizes MDM2  in the nucleoplasm [ UBTF codes for a protein playing critical roles in ribosomal RNA transcription and chromatin remodeling, which takes part in the compensatory response to proteotoxic stress in neurons [ ARS2 participates in maintaining neuronal stem cell identity via direct transcriptional activation of Sox2 [ PLA2G6, aliase PARK14, codes for a phospholipase A2, group 6 which hydrolases membrane phospholipids and may contribute, via lipid peroxidation, to CNS injury and disorders, such as Parkinson's disease [The VA-PD network  had, comc injury  and the c injury . SHARPINpoptosis –38. GNL3leoplasm  and promleoplasm . UBTF co neurons . ARS2 pa of Sox2 . Finally disease . Not sur disease .k values were 7.86 and 10.89 for LC-CT  and 164 genes and 645 gene-gene links (threshold 0.90) were obtained for LC-PD network and LC-CT network, respectively. or LC-CT  and LC-Por LC-CT .  Table  NUDT13, a common hub of both LC-CT and LC-PD networks, facilitates the elimination of oxidized forms of NAD(P)H and CoA cofactors from peroxisomes, mitochondria, and the cytoplasm [ SEPP1 codes for a brain antioxidant selenoprotein secreted by astrocytes and taken up by neurons via the apolipoprotein E receptor 2 [ GRM3, the metabotropic glutamate receptor 3 gene, and UGT8, UDP-glucuronosyltransferase 8, are closely interconnected hubs of the LC-CT network  and may cause neurodegenerative diseases depending on the length of polyQ expansions [In the LC-PD network all highly linked hubs are clustered in a single central module . Inside ic unity  and ATXNpansions , modulatpansions , 80. Trapansions .The remaining four genes in this set also exert relevant functions in neuroprotection and brain homeostasis, some of them possibly linked to repairing cellular injuries in PD, as discussed below. MED30, which codes for the mediator of polymerase II transcription subunit 30, participates in oxidative phosphorylation and mitochondrial integrity [ TOB2 regulates mRNA deacetylation, potentiates NGF-induced differentiation, and protects neurons from apoptosis [ SFRS18  codes for a serine-arginine rich protein which regulates protein sumoylation [α-synuclein aggregation and toxicity [ ZNFR3, a gene coding for a cell surface transmembrane E3 ubiquitin ligase zinc and ring finger 3, promotes Wnt receptor turnover [β-catenin signaling and antioxidant/anti-inflammatory pathways leads to the decline of subventricular zone (SVZ) plasticity with age and the limited nigrostriatal dopaminergic self-repair in PD [ ZNFR3 in PD would be necessary.The genentegrity . TOB2 repoptosis , 60. SFRoylation , a procetoxicity . Finallyturnover . This geturnover , (ii) returnover , and  and 199 genes and 682 gene-gene links (threshold 0.97) were obtained for SN-PD network and SN-CT network, respectively.  CBFB as a center, a gene coding for the beta subunit of a core-binding transcription factor belonging to the PEBP2/CBF transcription factor family which controls the transcriptional regulation of neurotrophin receptors, some ion channels, and neuropeptides, playing important roles in neuron development [In the control group network (SN-CT), the highly linked hubs were clustered in two modules . One modelopment , 92. The BCKDHB and SIRT1, were closely linked  and play important neuroprotective roles in the aging brain and PD [ SIRT1 deacetylates heat shock factor 1 (HSF1), increasing the transcription of molecular chaperones such as heat shock protein 70. SIRT1 interacts with peroxisome proliferator-activated receptor-gamma coactivator-1α (PGC-1α) to reduce oxidative stress and increase the viability of dopaminergic neurons [ SIRT1 may also regulate autophagy and mitophagy, which may diminish α-synuclein toxicity in PD [ SORT1 (sortilin1), a common hub of SN-CT and SN-PD networks. Sortilin is a member of the family of vacuolar protein sorting 10 protein domain receptors. It is a coreceptor in cell death and neurodegeneration processes mediated by proneurotrophins (proNT) [ SIRT1 expression is altered in the aging brain and aged neurons are more sensitive to proNT-induced killing than young ones [Two hubs,y linked  and playeostasis , 90. SIRn and PD . SIRT1 d neurons . Moreovety in PD . Interes (proNT) , 102 and (proNT) . It is wung ones . Consequ ZFP112, a gene coding for an unknown zinc-finger protein, and SHC4 and TMEM123, both having relevant roles in neuronal homeostasis. SHC4 codes for a Src homology and collagen (Shc) protein that interacts with tropomyosin receptor kinase B, trkB [ TMEM123 codes for transmembrane protein 123, a cell surface receptor mediating oncolytic cell death [Three other hubs are B, trkB , the hig B, trkB . TMEM123ll death . Oncosisll death , and it ll death . Altoget BCKDHB and SORT1 (previously discussed), are common to both SN-CT and SN-PD networks. The most connected hub in SN-PD network  and neuronal cell adhesion molecule (NrCAM) in the molecular assembly of the nodes of Ranvier in the peripheral nervous system [ GLDN expressed in SN cells could have a role in these processes. Furthermore, diffusion tensor imaging studies showed significant regional (substantia nigra) and global white matter deterioration in PD [Conversely, the SN-DP network showed a significant change in the modular structure with all the highly connected hubs clustered in a single module . Two of  network  was CLDNcompacta  and invocompacta . In factcompacta . The secs system , 95. Sins system , it is ron in PD . ARID4B, MBTD1, and HNRNPA3, have been shown to participate in biological processes associated with neurodegeneration. ARID4B is a chromatin remodeling gene coding for a protein associated with the mSIN3A histone deacetylase (HDAC) complex [ MBTD1 is a member of the Polycomb gene family [ HNRNPA3 codes for a shuttling RNA transporter found in the neuronal RNA granules and P-bodies [ SIRT1, a SN-CT hub, exerts its neuroprotective action by inhibiting the ARID4B/mSINA3/HDAC transcriptional repression activity [ SIRT1 activity gets apparently attenuated in the SN-PD network.The last three SN-PD hubs, complex , which p complex . MBTD1 ie family , and itse family , thus coe family . In PD, e family . HNRNPA3P-bodies . These sP-bodies , 116. Heactivity . This SIThe SN-PD transcriptional network profile, with a predominance of hubs linked to neurodegenerative processes , would be expected to be found in the SN of patients in Braak stages 4-5 , 118–120α-synuclein molecules [Solid evidences now exist that PD has a caudal-rostral progression, being initiated in the dorsal motor vagal nucleus and/or olfactory bulb, progressing to the midbrain and eventually to the basal forebrain and the neocortex , 5, 6 tholecules , 8, 121.olecules , 5, 6, solecules , 121. Itolecules .This report shows that transcriptional interaction network analysis, an effective methodology for dealing with a large set of genomic data , 19, 20,2+ homeostasis, myelination, and neuroprotection in the aging brain, whereas, in VA-PD, the relevant hubs are mostly related to compensatory responses to proteotoxic stress. Interestingly, one of these hubs is PARK14 , a gene causing autosome recessive early-onset PD when mutated [SFRS18, ZFNR3, and FAM5B, for instance). Finally, the SN-CT main hubs include genes that are critical for neuroprotection and homeostasis in the aging brain, such as SIRT1 and BCKDHB. Conversely, the SN-PD network displays a very different landscape: six out seven hubs are associated with neurodegenerative processes. These results are compatible with the caudo-rostral model of PD progression and point out to the usefulness of GCNs approach for (i) investigating the molecular mechanisms underlying idiopathic PD and (ii) identifying novel therapeutic targets based on the concept of interventions aimed to restore altered regulatory network structures [In VA-CT, the main hubs are associated with Ca mutated . The LC-ructures , 20, 122


Cancer is one of the most life-threatening diseases characterized by uncontrolled growth and spread of malignant cells. The tumor suppressor p53 is the master regulator of tumor cell growth and proliferation. In response to various stress signals, p53 can be activated and transcriptionally induces a myriad of target genes, including both protein-encoding and non-coding genes, controlling cell cycle progression, DNA repair, senescence, apoptosis, autophagy and metabolism of tumor cells. However, around 50% of human cancers harbor mutant p53 and, in the majority of the remaining cancers, p53 is inactivated through multiple mechanisms. Herein, we review the recent progress in understanding the molecular basis of p53 signaling, particularly the newly identified ribosomal stress—p53 pathway, and the development of chemotherapeutics via activating wild-type p53 or restoring mutant p53 functions in cancer. A full understanding of p53 regulation will aid the development of effective cancer treatments. This could be the reason why the mutant, but not wild-type, p53 was firstly cloned from cancer cells. To date, tens of thousands of studies have demonstrated that p53 expands its tumor suppressive functions to most, if not all, aspects of cancer development. p53 is a transcriptional factor that regulates a large number of target genes involved in cell cycle arrest, apoptosis, senescence, autophagy and metabolism [N- and C-termini of p53 [N-terminal transactivation domain [p53 gene rescues the lethal phenotype of Mdm2 knock-out mice [Tumorigenesis is promoted by deregulation or mutation of genes involved in cell division control. The tumor suppressor p53 is the most prominent and extensively studied example due to its dominating effect on tumor cell growth and proliferation ,2,3,4,5.pathways ,7,8. Althe 1980s . Baker e [et al.  found thtabolism   has emerged and been gradually appreciated as an essential cellular pressure that can induce p53 activation and resultant tumor growth inhibition ,45. Riboin vivo study has gracefully validated the physiological significance of RPs in ribosomal stress-triggered p53 activation. In this study, Macias et al. [C305F which was defective in binding to RPL5 and RPL11, and found that p53 activation in response to ribosomal stress was markedly impaired, whereas p53 response to DNA damage was normal, in these mutant mice [Although the association of RPL5 and MDM2 was initially identified in 1994 , the phys et al. ,67 generant mice .et al. [Ganoderma lucidum [1 cell cycle arrest and suppress proliferation of lung cancer cells by triggering ribosomal stress and RPS7-MDM2 binding [Growing evidence has demonstrated the central role of the RP-MDM2-p53 pathway in controlling cancer development. For instance, Sasaki et al.  found thet al. . Anotheret al. . In addiet al. . ARF canet al. . Ling Zh lucidum  can indu binding . The aboInterestingly, post-translational modifications (PTM) of RPs have been found to be essential for MDM2-p53 regulation. Under normal conditions, the NEDD8-mediated RPL11 NEDDylation promotes its nucleolar localization, whereas ribosomal stress can induce de-NEDDylation of RPL11 which subsequently relocalizes to the nucleoplasm leading to p53 activation ,73. In cTaken together, this gradually appreciated RP-MDM2-p53 pathway is subjected to multiple regulations, including RP-associating cofactors and PTMs, and has been playing an increasingly essential role in controlling tumorigenesis.etc. [It has been well documented that most p53 functions are mediated by transcriptional activation of its target genes in response to various stress signals , althougetc. ,29,81. IPUMA, p53AIP1 and NOXA, also have high-affinity p53 REs [p53AIP1, and Ser-46 to Alanine mutation that prevents phosphorylation of this site abolishes p53-mediated activation of p53AIP1, but not the cell cycle gene p21 [p21, but fails to induce the expression of the apoptotic target genes, PUMA and BAX [What accounts for p53 selective activation of its target genes dictating diverse cell fates is an intriguing question. A number of studies have demonstrated factors that regulate target gene selection by p53 . Several p53 REs . Hence, gene p21 . Another and BAX ,39. Addi and BAX . The ass and BAX ,89. AltoAside from the aforementioned protein-encoding genes, p53 can also induce non-coding RNA genes, including microRNA (miRNA) and long non-coding RNA genes. miRNAs are small noncoding RNAs, 9–22 nucleotides in length, which usually bind to the 3' UTR of target mRNAs and post-transcriptionally silence their expression. Principally, the generation of a functional miRNA includes two steps, transcription and maturation. RNA Pol II or RNA Pol III-mediated transcription produces the miRNA precursor, pri-miRNA, which undergoes further processing to generate mature miRNA. Only the mature miRNA is employed to form the RNA-induced silencing complex (RISC) to inhibit the target gene expression . InteresWhen talking about wild-type p53, it is impossible not to mention mutant p53. Most cancer-associated inactivation of p53 results from missense mutations, single base-pair substitutions leading to the translation of a different amino acid that may either annihilate the wild-type p53 function or endow the mutant p53 with new functions. A seminal study has recently found that the elevated expression of the DNA cytosine deaminase APOBEC3B in breast cancer may contribute to the mutagenesis of p53 . BecauseAs the essential role of p53 is killing tumor cells, numerous strategies have been developed to activate p53 or restore mutant p53 function for cancer therapy. Although some traditional genotoxic chemotherapeutic drugs, such as Adriamycin and platinum-based drugs, activate the p53 pathway, they also cause systemic toxicity and induce multidrug resistance. It is therefore critically important that non-genotoxic chemotherapeutics be developed to specifically target p53. Because of the central role of MDM2 in modulating p53 activity, much attention has been devoted to developing MDM2 inhibitors that can specifically release p53 from MDM2 repression. The ground-breaking work in this field is the development of Nutlin, a small molecule disrupting MDM2-p53 interaction without causing genotoxicity . It has One of the important negative regulators, the SIRT1 deacetylase, has been shown to maintain the deacetylated status of p53, and thus promote its MDM2-mediated degradation ,128. It MET . While PRIMA-1MET can induce cell death independently of p53 [in vitro and in vivo by covalently binding to mutant p53 and mediating modification of its thiol groups, and has completed a Phase I clinical trial [R175 mutant into a wild-type structure by chelating the zinc ion and changing the redox state, and therefore rescues transcriptional function of p53R175 mutant resulting in tumor suppression [As mutations in p53 occur in up to 50% of human cancers, how to restore mutant p53 protein to a transcriptionally functional conformation becomes a hot topic and promising strategy for the treatment of cancers harboring mutant p53. One of the most successful compounds is PRIMA-1y of p53 ,132,133,al trial ,136,137.al trial ,139. Theal trial ,139. Recpression .There are certainly many other compounds, in addition to those mentioned above, targeting p53 or the mutant p53 pathway ; however3KR was shown to partially lose its transcriptional activity leading to abrogated cell cycle arrest, apoptosis and senescence upon DNA damage while retaining tumor suppressive function by inducing a range of target genes involved in metabolic regulation [As one of the most important tumor suppressors, p53 can be activated in response to multiple stress signals leading to transcriptional activation of a large group of target genes and consequently suppression of tumorigenesis principally through inducing cell cycle arrest, senescence, apoptosis and metabolic changes. Therefore, the p53 pathway has become an ideal target for the development of chemotherapeutics for cancer. While the achievements in dissecting the p53 signaling pathway opens a promising avenue which might lead to our final success in the battle against cancer, these seminal studies also raise a great many questions. For example, acetylation of several lysine residues, such as Lys-120 and Lys-164, are critical for transcriptional activity of p53. To assess whether full activity of p53 is essential for its tumor suppressive ability, an animal model with triple lysine to arginine mutations of Lys-117, Lys-161 and Lys-162  was generated . In the gulation . Anothergulation ,145. The


Citrus is a kind of common fruit and contains multiple beneficial nutrients for human beings. Flavonoids, as a class of plant secondary metabolites, exist in citrus fruits abundantly. Due to their broad range of pharmacological properties, citrus flavonoids have gained increased attention. Accumulative in vitro and in vivo studies indicate protective effects of polymethoxyflavones (PMFs) against the occurrence of cancer. PMFs inhibit carcinogenesis by mechanisms like blocking the metastasis cascade, inhibition of cancer cell mobility in circulatory systems, proapoptosis, and antiangiogenesis. This review systematically summarized anticarcinogenic effect of citrus flavonoids in cancer therapy, together with the underlying important molecular mechanisms, in purpose of further exploring more effective use of citrus peel flavonoids. The citrus flavonoids include a class of glycosides, namely, hesperidin and naringin, and another class of O-methylated aglycones of flavones such as nobiletin and tangeretin, which are relatively common two polymethoxylated flavones (PMFs) [In our daily diet, the average intake of flavonoids of every day ranges from 150 mg to 300 mg . The mais (PMFs) , and ants (PMFs) . Meanwhis (PMFs) , diabetes (PMFs) , and neus (PMFs) .In recent years, epidemiological studies have shown that there is a connection that flavonoid intake may reduce the risk of developing colon cancer , 12. MorCancer is the life threatening and dreadful disease characterized by the abnormal proliferation of cells that invade the adjacent tissues and cause the destruction of these tissues. It is the second leading cause of death all over the world. More than six million deaths each year occurring in the world are due to cancer. Several lines of evidence indicated that tumorigenesis in humans is a multistep process and these steps reflect genetic alterations that drive the progressive transformation of normal human cells into highly malignant derivatives . ConventIn the tumor microenvironment, from cancer cells initiation to promotion and eventually progression, compelling evidence indicates the potential activities of flavonoids in citrus peel cover inhibiting oncogenesis, proliferation, neovascularization, and metastasis and inducing apoptosis.  Citrus aurantium L. [Cip1/Waf1 and downregulating cdc2 and cyclin B1 [Cell cycle abnormalities are closely associated with cancer, and citrus peel flavonoids substantially influence on cell cycle arrest. Cell cycle is an important regulatory mechanism of cell growth, development, and differentiation. In mammals, the cell cycle comprises the G1, S, G2, and M phases. Cell cycle checkpoints keep the maintenance of genomic integrity by inhibiting damaged or incomplete DNA. G2/M checkpoint ensures that the cells do not initiate mitosis before repairing damaged DNA after replication. The cell cycle progression depends on a cascade of enzymes by sequential activation and inactivation of cyclin, cyclin-dependent kinases (CDKs), and cyclin-dependent kinase inhibitors (CDKIs) . Cdc2 inntium L. . Downregntium L. . It is antium L. , 23. In ntium L. . More reyclin B1 . Among tyclin B1 . The che2+ and mobilization of intracellular Ca2+ store, accompanied by activation of calpain and caspase-12 [ Citrus aurantium L. induced caspase-dependent apoptosis through Akt pathway by inhibiting expression of XIAP and Bcl-2 which are antiapoptotic proteins, providing the fact that they have anticarcinogenic activity on human leukemia cells U937 [γ) and suppressing the activation of NF-κB [50 values with increasing concentration of OH-PMFs were observed in different orange peel extracts [2+ were lower than those of nonhydroxylated PMFs [One of the most basic features of cancer cells is their ability to proliferate chronically. Apart from blocking cell cycle, flavonoids in citrus peel can also inhibit cell proliferation and promote apoptosis, especially in triple-negative (ER-/PR-/HER2-) breast cancer (TNBC). PMFs triggered influx of Caspase-12 . There alls U937 . In anotof NF-κB . Tangereof NF-κB . In HCT1of NF-κB . Noteworextracts . Subsequted PMFs . These aκB into the nucleus [Kip2 [In the research of mice, oral feeding of gold lotion (GL), a formulated product made from the peels of six citrus fruits, decreased the number of aberrant crypt foci (ACF) in mice colonic tissues . This co nucleus . Similar nucleus . Moreoveus [Kip2 . Data abψm dissipation, release of cytochrome C, and cleavage of caspase-9 after exposure to 5-ATAN in a time-dependent manner which indicated caspase-dependent intrinsic pathway were also required in the MCF-7 cells [Our latest research showed that, in MCF-7 human breast cancer cells, 5-acetyl-6,7,8,4′-tetramethylnortangeretin (5-ATAN), which replaces the methyl groups of tangeretin with acetyl groups at the C5 position of tangeretin, showed more powerful abilities than its parent compound. Then, we looked for evidence of 5-ATAN on apoptosis. Translocation of apoptosis-inducing factor (AIF) and phosphorylation of H2AX are commonly used for evaluating the impact of natural compounds-induced caspase-independent apoptosis pathway , 38. Our-7 cells . All theμM hesperidin together in treating with MCF-7 cells, they increased cytotoxic effect, modulated cell cycle, and induced apoptosis of MCF-7 cells [Traditional treatment of cancer has been facing a huge number of problems, in view of its complex molecular pathophysiology that varies according to each type. Several ways in the treatment of breast cancer have been developed that are surgery, chemotherapy, hormonal therapy, and radiation. Doxorubicin, a chemotherapeutic agent commonly used in breast cancer treatment, showed low effectivity, rendering its resistance and toxicity on normal tissues . An appr-7 cells . Meanwhi-7 cells . Cycloph-7 cells .α [Invasion and metastasis are a multistep process and are described as a series of discrete steps, usually called invasion—the metastatic cascade . It descα . Besidesα .Angiogenesis is a physiological process of forming new blood vessels from preexisting vessels, which involves the induction of new sprouts, coordinated and directed endothelial cell migration, proliferation, sprout fusion, and lumen formation . Similar VEGF-A gene encodes the ligand involved in neovascularization during the embryonic and neonatal development, homeostasis, and survival of endothelial cells, as well as physiological and pathological state of adult [As one of the angiogenesis inducers, vascular endothelial growth factor-A (VEGF) can be used as a marker of angiogenesis.of adult . When miof adult . In addiof adult .50 value being 10 lg (24.9 nmol) per egg [Through combined inhibition of multiple angiogenesis-related endothelial cells (EC) functions, nobiletin had been demonstrated to have concentration-dependent inhibitory effects on angiogenic differentiation induced by VEGF and FGF (fibroblast growth factors). In a chick embryo chorioallantoic membrane assay, nobiletin showed an antiangiogenic activity with the ID per egg . flt1, kdrl, and hras in zebrafish [With human umbilical vein endothelial cells (HUVECs) in vitro and zebrafish in vivo models, PMFs showed different degrees of potency of antiangiogenesis activity. Sinensetin, which showed the most potent antiangiogenesis activity and the lowest toxicity, inhibited angiogenesis by inducing cell cycle arrest in the G0/G1 phase in HUVEC culture and downregulating the mRNA expressions of angiogenesis genesebrafish . Nobiletebrafish , structuebrafish .Flavonoids also exert their chemopreventative effect via inhibition of certain phase I metabolizing enzymes, such as cytochrome P450 which metabolically activates a large number of procarcinogens triggering carcinogenesis. The chemopreventative effects of flavonoids are closely linked to their anticancer properties that involve the scavenging of reactive oxygen species (ROS) and growth promoting oxidants which are the major catalysts for tumor promotion. Tangeretin, a polymethoxylated flavone, can inhibit cancer cell proliferation by improving antioxidant properties such as decreasing the levels of lipid peroxide, enzymatic antioxidants SOD, CAT, and GPx, and nonenzymatic antioxidants such as GSH, vitamin C, and vitamin E in 7,12-dimethyl benz(a)anthracene (DMBA) induced mammary carcinoma in rats . OtherwiPharmacokinetics describes how the body affects a specific drug after administration through mechanisms of absorption and distribution, as well as the chemical changes of the substance in the body. At a practical level, a drug's bioavailability can be defined as the proportion of the drug that reaches its site of action. Poor absorption and extensive conjugative metabolisms greatly limit bioavailability of dietary flavonoids.The bioavailability is an overall effect of absorption, distribution, metabolism, and excretion and plays an important role in dictating cancer preventive efficacy of dietary components in humans. Bioavailability testing can be divided into in vitro and in vivo bioavailability. In vitro bioavailability test can be a good predictor of the latter one.Currently, human colon adenocarcinoma cell line caco-2 cell model is established to simulate the human intestinal absorption so as to test permeability and study absorption mechanism. The caco-2 data of 3′-hydroxy-5,6,7,4′-tetramethoxyflavone, 3,5,6,7,8,3′,4′-heptamethoxyflavone, and 3-hydroxy-5,6,7,8,3′,4′-hexamethoxyflavone showed superb permeability . Meanwhi50 value of 5-demethyltangeretin (5DT) was 78.9-fold lower than that of tangeretin. Since cancer cells can pump the cytotoxic agents out via overexpression of multidrug resistant efflux proteins, cells were treated with 5DT or tangeretin at the same concentration. HPLC analysis revealed that the intracellular levels of 5DT in NSCLC cells were 2.7–4.9-fold higher than those of TAN. This suggested that NSCLC cells may have better uptake efflux of 5DT compared with TAN. Additionally, molecular structure showed that 5DT had higher lipophilicity than tangeretin [Also, in NSCLC A549 cell line, 5-hydroxylated PMFs had much stronger inhibitory effects on cancer cells in comparison with their permethoxylated counterparts, for ICngeretin . High li50 of 0.12, 5.5, and 4.2 μM in SW620 cells, while 5-demethylnobiletin at 10 μM only caused 37% inhibition [Biotransformation of dietary components is crucial for their in vivo biological activities after oral ingestion because the process of drug metabolism notably influences drugs effects and toxicity. In the research of nobiletin metabolites, by comparing supercritical fluid chromatography (SFC) profiles of metabolite mixtures with the synthesized standard compounds, three major metabolites were proved to be 4′-demethylnobiletin, 3′-demethylnobiletin, and 3′,4′-didemethylnobilietin in mouse urine . Furtherhibition . Hence, Taken all together, a considerable number of well-established lines of evidence have confirmed that flavonoids in citrus peel exhibit a remarkable spectrum of efficacious biological activities, particularly in antitumorigenesis. Excellent permeability through membrane allows citrus flavonoids to possess great bioavailability which consequently attracts researchers to perform scientific studies for effective disease prevention and treatment. There are more modified flavonoids in citrus peel being investigated, which could offer help to improve dose-effect relationship greatly and advance the security and stability of compounds.