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Rotundic acid (RA), an ursane-type pentacyclic triterpene acid isolated from the dried barks of Ilex rotunda Thunb. (Aquifoliaceae), possesses diverse bioactivities. To further study its pharmacokinetics, a simple and sensitive liquid chromatography with triple quadrupole mass spectrometry (LC-QqQ-MS/MS) method was developed and validated to quantify RA concentration in rat plasma and tissue using etofesalamide as an internal standard (IS). Plasma and tissue samples were subjected to one-step protein precipitation. Chromatographic separation was achieved on a ZORBAX Eclipse XDB-C18 column (4.6 mm × 50 mm, 5 μm) under gradient conditions with eluents of methanol:acetonitrile (1:1, V/V) and 5 mM ammonium formate:methanol (9:1, V/V) at 0.5 mL/min. Multiple reaction monitoring transitions were performed at m/z 487.30 → 437.30 for RA and m/z 256.10 → 227.10 for IS in the negative mode. The developed LC-QqQ-MS/MS method exhibited good linearity (2–500 ng/mL) and was fully validated in accordance with U.S. Food and Drug Administration bioanalytical guidelines. Dose proportionality and bioavailability in rats were determined by comparing pharmacokinetic data after single oral (10, 20, and 40 mg/kg) and intravenous (10 mg/kg) administration of RA. Tissue distribution was studied following oral administration at 20 mg/kg. The results showed that the absolute bioavailability of RA after administration at different doses ranged from 16.1% to 19.4%. RA was rapidly absorbed in a dose-dependent manner and highly distributed in the liver. In conclusion, this study is the first to systematically elucidate the absorption and distribution characteristics of RA in rats, which could provide additional information for further development and evaluation of RA in drug metabolism and pharmacokinetic studies.

Objective Osteosarcoma is one of the most common types of bone sarcoma with a poor prognosis. However, identifying the predictive factors that contribute to the response to neoadjuvant chemotherapy remains a significant challenge. Methods A public data series (GSE87437) was downloaded to identify differentially expressed genes (DEGs) and differentially expressed lncRNAs (DElncRNAs) between osteosarcoma patients that do and do not respond to preoperative chemotherapy. Subsequently, functional analysis of the transcriptome expression profile, regulatory networks of DEGs and DElncRNAs, competing endogenous RNAs (ceRNA) and protein-protein interaction networks were performed. Furthermore, the function, pathway, and survival analysis of hub genes was performed and drug and disease relationship prediction of DElncRNA was carried out. Results A total of 626 DEGs, 26 DElncRNAs, and 18 hub genes were identified. However, only one gene and two lncRNAs were found to be suitable as candidate gene and lncRNAs respectively. Conclusion The DEGs, hub genes, candidate gene, and candidate lncRNAs screened out in this context were considered as potential biomarkers for the response to neoadjuvant chemotherapy of osteosarcoma.

Structures of BbKI, a recombinant Kunitz-type serine protease inhibitor fromBauhinia bauhinioides, complexed with human kallikrein 4 (KLK4) were determined at medium-to-high resolution in four crystal forms (space groupsP3121,P6522,P21andP61). Although the fold of the protein was virtually identical in all of the crystals, some significant differences were observed in the conformation of Arg64 of BbKI, the residue that occupies the S1 pocket in KLK4. Whereas this residue exhibited two orientations in the highest resolution structure (P3121), making either a canonical trypsin-like interaction with Asp189 of KLK4 or an alternate interaction, only a single alternate orientation was observed in the other three structures. A neighboring disulfide, Cys191–Cys220, was partially or fully broken in all KLK4 structures. Four variants of BbKI in which Arg64 was replaced by Met, Phe, Ala and Asp were expressed and crystallized, and their structures were determined in complex with KLK4. Structures of the Phe and Met variants complexed with bovine trypsin and of the Phe variant complexed with α-chymotrypsin were also determined. Although the inhibitory potency of these variant forms of BbKI was lowered by up to four orders of magnitude, only small changes were seen in the vicinity of the mutated residues. Therefore, a totality of subtle differences in KLK4–BbKI interactions within the fully extended interface in the structures of these variants might be responsible for the observed effect. Screening of the BbKI variants against a panel of serine proteases revealed an altered pattern of inhibitory specificity, which was shifted towards that of chymotrypsin-like proteases for the hydrophobic Phe and Met P1 substitutions. This work reports the first structures of plant Kunitz inhibitors with S1-family serine proteases other than trypsin, as well as new insights into the specificity of inhibition of medically relevant kallikreins.

The forest tree family Aceraceae is widespread in the northern hemisphere and it has ecological and economic importance. However, the phylogenetic relationships and classifications within the family are still controversial due to transitional intraspecific morphological characteristics and introgression hybridization among species. In this study, we determined the evolutionary relationships and molecular evolution of Aceraceae based on plastid phylogenomics and two nuclear gene variations. Phylogenetic analysis based on the plastid genomes suggested that Aceraceae species can be divided into two larger sub-clades corresponding to the two genera Acer and Dipteronia. Conjoint analysis of the plastid and nuclear gene sequences supported the classification with two genera in the family. Molecular dating showed that the two genera diverged 60.2 million years ago, which is generally consistently with previously reported results. Divergence hotspots and positively selected genes identified in the plastid genomes could be useful genetic resources in Aceraceae.

Background: Competing endogenous RNA (ceRNA) networks play a pivotal role in tumor diagnosis and progression. Numerous studies have explored the functional landscape and prognostic significance of ceRNA interaction within differentiated tumor cells. Objective: We propose a new perspective by exploring ceRNA networks in the process of glioblastoma stem cell (GSC) differentiation. Methods: In this study, expression profiles of lncRNAs and mRNAs were compared between GSCs and differentiated glioblastoma cells. Using a comprehensive computational method, miRNAmediated and GSC differentiation-associated ceRNA crosstalk between lncRNAs and mRNAs was identified. A ceRNA network was then established to select potential candidates that regulate GSC differentiation. Results: Based on the specific ceRNA network related to GSC differentiation, we identified lnc MYOSLID: 11 as a ceRNA that regulated the expression of the downstream gene PXN by competitively binding with hsa-miR-149-3p. After Kaplan-Meier (KM) survival analysis, the expression of PXN gene (PPXN = 0.0015) and lnc MYOSLID: 11 (PMYOSLID: 11=0.041) showed significant correlation with glioblastoma in 160 patients from TCGA. Conclusion: This result sheds light on a potential way of studying the ceRNA network, which can provide clues for developing new diagnostic methods and finding therapeutic targets for clinical treatment of glioblastoma.

A novel Gram-negative, motile, aerobic rod-shaped bacterium designated BGMRC 2031T was isolated from mangrove sediment collected from Guangxi Province, China. Optimal growth occurred at 28 °C and pH 7.0–8.0 in the presence of 1% (w/v) NaCl. Alignment based on 16S rRNA gene sequences indicated that strain BGMRC 2031T is most closely related to Sodalis praecaptivus HS1T (95.6%, sequence similarity), followed by Biostraticola tofi DSM 19580T (95.5%), Sodalis glossinidius DSM 16929T (95.4%), and Brenneria goodwinii FRB141T (94.9%) sequence similarity. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain BGMRC 2031T formed a distinct branch in a robust cluster and revealed that strain BGMRC 2031T, genera Biostraticola and Sodalis, formed a novel family-level clade in the order Enterobacterales. The novel strain showed an average nucleotide similarity of 74.7%, 74.2%, and 73.1% for S. praecaptivus HS1T, S. glossinidius DSM 16929T, and B. tofi DSM 19580T, respectively. The genomes of the BGMRC 2031T shared the presence of a riboflavin synthesis gene cluster. The menaquinones of strain BGMRC 2031T were MK-8 and Q-8, which were similar to those of genus Biostraticola. The major fatty acids (> 10%) were C16:0 (19.9%), summed feature 2 (iso-C16:1 and/or C14:0 3-OH, 18.10%), summed feature 3 (C16:1ω7c and/or C16:1ω6c, 15.3%), C12:0 (13.9%), C17:0 cyclo (11.4%), and C14:0 (10.4%). The main polar lipids were phosphatidyl methylethanolamine, phosphatidyl glycerol, diphosphatidyl glycerol, phosphatidyl inositol, one unidentified phospholipid, and one unknown polar lipid. The G+C content of strain BGMRC 2031T was 55.4%. Strain BGMRC 2031T could extend the mean lifespan and maximum lifespan of Caenorhabditis elegans by 4.5% and 12.5%, respectively. Overall, the results of this study indicate that BGMRC 2031T is a novel species in a new genus, for which the name Bruguierivorax albus gen. nov. sp. nov. is proposed, and the type of strain is designated as BGMRC 2031T (= NBRC 111907T = KCTC 52119T). In addition, a novel family, Bruguierivoracaceae fam. nov., is proposed to accommodate the genera Bruguierivorax, Biostraticola, and Sodalis.

Illihenin A (1), a novel sesquiterpenoid, was isolated from the roots of Illicium henryi. The structure was determined by spectroscopic analyses, ECD calculation, and single-crystal X-ray diffraction. Compound 1 represents a class of novel 5/7/6 tricyclic sesquiterpenoids featuring a rare cage-like tricyclo[6.2.2.01,5]dodecane core. A plausible biosynthetic pathway of 1 by rearrangement of allo-cedrane is proposed. Additionally, 1 showed potent antiviral activity against coxsackievirus B3 with an IC50 value of 2.87 μM.

Objective: Based on the GEO database, the differential genes of sinusoidal endothelial cells in cirrhotic rats were analyzed. Methods: In the GEO database, the differential gene expressions of liver sinusoidal endothelial cells in cirrhotic rats were obtained. The screening was performed according to P < 0.01 and differential multiple factor ≥ 4. The obtained genes were input into the DAVID database for enrichment analysis of genes and pathways. The GeneMania and string databases were then used for protein interaction analysis. Results: The GSE1843 dataset was obtained in the GEO database, and three pathways significantly associated with cirrhosis and 13 differential genes enriched in three pathways were screened. Text mining revealed that 11 differential genes were directly associated with cirrhosis. The other two were indirectly linked by other genes. The screened genes and known gene formation networks were discovered by the GeneMania tool. Conclusion: CDH2 and COL1A1 may be important target genes for cirrhosis.

Upon sensing cytosolic DNA, the enzyme cGAS induces innate immune responses that underpin anti-microbial defenses and certain autoimmune diseases. Missense mutations of PRKDC encoding the DNA-dependent protein kinase (DNA-PK) catalytic subunit (DNA-PKcs) are associated with autoimmune diseases, yet how DNA-PK deficiency leads to increased immune responses remains poorly understood. In this study, we report that DNA-PK phosphorylates cGAS and suppresses its enzymatic activity. DNA-PK deficiency reduces cGAS phosphorylation and promotes antiviral innate immune responses, thereby potently restricting viral replication. Moreover, cells isolated from DNA-PKcs-deficient mice or patients carrying PRKDC missense mutations exhibit an inflammatory gene expression signature. This study provides a rational explanation for the autoimmunity of patients with missense mutations of PRKDC, and suggests that cGAS-mediated immune signaling is a potential target for therapeutic interventions., The enzyme cGAS induces innate immune responses upon recognition of cytosolic DNA. Here, using in vitro and in vivo models, the authors identify DNA-PK as a negative regulator of cGAS signalling.

The survival benefit of sorafenib, the most used drug for advanced hepatocellular carcinoma (HCC), is unsatisfactory due to the development of adaptive resistance. Exploring the mechanisms underlying sorafenib resistance is important to develop sensitizing strategy. Sphingomyelin synthase (SMS) plays a critical role in sphingolipid metabolism which is involved in oncogenesis and drug resistance. SMS1 and SMS2 levels in HCC cells in response to prolonged chemotherapy were analyzed using ELISA. mRNA and protein levels of SMS in HCC and adjacent normal tissues were analyzed by ELISA and real-time PCR. The roles of SMS and its downstream targets were investigated using cellular and biochemical assays and mass spectrometry. SMS1, but not SMS2, was upregulated in HCC in response to sorafenib treatment, although HCC displayed similar RNA and protein level of SMS1 compared to adjacent normal liver tissues. Overexpression of SMS1 promoted HCC growth and migration, and alleviated sorafenib’s toxicity. SMS1 inhibition via genetic and pharmacological approaches consistently resulted in inhibition of growth and migration, and apoptosis induction in sorafenib-resistance HCC cells. SMS1 inhibition also augmented the efficacy of sorafenib in sensitive HCC cells. SMS1 inhibition disrupted sphingolipid metabolism via accumulating ceramide and decreasing sphingomyelin, inducing mitochondrial dysfunction and oxidative stress, and decreasing Ras activity in resistant cells. Overexpression of constitutively active Ras reversed the inhibitory effects of SMS1 inhibition. Although SMS1 overexpression did not affect Ras expression and activity, Pearson correlation coefficient analysis of SMS1 and Ras expression demonstrated that there was positive correlation between SMS1 and RAS (NRAS, R = 0.55, p

Geological and climatic events are considered to profoundly affect the evolution and lineage divergence of plant species. However, the evolutionary histories of tree species that have responded to past geological and climate oscillations in central China's mountainous areas remain mostly unknown. In this study, we assessed the evolutionary history of the endangered and relict tree species Dipteronia sinensis in the Qinling Mountains (QM) and adjacent areas in East Asia based on variations in the complete chloroplast genomes (cpDNA) and reduced‐genomic scale single nucleotide polymorphisms (SNPs). Population structure and phylogenetic analysis based on the cpDNA variations suggested that D. sinensis could be divided into two intraspecific genetic lineages in the eastern and western sides of the QM (EQM and WQM, respectively) in East Asia. Molecular dating suggested that the intraspecific divergence of D. sinensis occurred approximately 39.2 million years ago during the later Paleogene. It was significantly correlated with the orogeny of the QM, where the formation of this significant geographic barrier in the region may have led to the divergence of independent lineages. Bayesian clustering and demographic analysis showed that intraspecific gene flow was restricted between the EQM and WQM lineages. Isolation‐with‐migration analysis indicated that the two genetic lineages experienced significant demographic expansions after the Pleistocene ice ages. However, the genetic admixture was determined in some populations between the two lineages by the large scale of SNP variations due to DNA incompatibility, the large significant population size, and rapid gene flow of nuclear DNA markers. Our results suggest that two different conservation and management units should be constructed for D. sinensis in the EQM and WQM areas. These findings provide novel insights into the unprecedented effects of tectonic changes and climatic oscillations on lineage divergence and plant population evolution in the QM and adjacent areas in East Asia., This novel study describes the unprecedented details of geological and climatic changes profoundly shaped the lineage divergence and population evolution of Dipteronia sinensis plant species in the Qinling Mountains in central China. The readers will take help to understand the mechanism of lineage divergence due to the mountain's orogenesis and repeated climatic changes.

Purpose: Low Molecular Weight Cyclin E (LMW-E) are the oncogenic forms of cyclin E and were originally discovered in breast cancer (BC). LMW-E are generated from N-terminal cleavage of the 50 KDa, full-length cyclin E1 (FL-cycE). Our laboratory has established that LMW-E expression (i) correlates with poorer survival in BC patients; (ii) increases in frequency as BC progress from ductal carcinoma in situ to invasive ductal carcinoma; (iii) drives spontaneous and metastatic BC in murine transgenic models; (iii) causes human mammary epithelial cells (hMECs) to transform in vitro and form tumors in vivo. Elucidating the critical oncogenic functions and essential downstream factors of LMW-E is a current void in the field and may have important therapeutic implications. Experimental Design: Immunohistochemistry (IHC) analysis were performed using tumor specimens (n=725) from breast cancer patients diagnosed with Stage 1 or 2 disease to determine the level of LMW-E. Micro-dissected tumor DNA from these tissues were analyzed for copy number variations (CNVs) using Molecular Inversion Probe (MIP) based arrays. CCNE1(encoding Cyclin E1) knock-out 76NE6 and 76NF2V hMEC lines were generated by CRISPR, followed by transfection of lentivirus vector expressing doxycycline inducible EGFP fused LMW-E or FL-cycE. Time-lapse live cell imaging was performed to monitor the cell growth and phenotypes after induced expression of LWM-E or FL-cycE. DNA replication and replication stress were examined by BrdU labeling and immunofluorescent (IF) assays using antibodies against BrdU and Replication Protein A (RPA). IF for gamma-H2AX and western blotting for phosphor-RPA32, ATR-CHK1 and ATM-CHK2 pathways were performed to determine the DNA damage and responses. Results: Analysis of CNVs for the 725 tumors stratified by the cyclin E phenotypes reveal that the frequency of the CNVs (gains & losses) were the most significant in patients whose tumors expressed LMW-E and were predictive of poor prognosis independent of BC subtypes. DNA damage signals including gamma-H2AX foci and phosphor-CHK1 were similarly enhanced in hMECs induced for FL-cycE or LMW-E over-expression. Different effects of FL-cycE and LMW-E on DNA replication and replication stress were observed. Induced FL-cycE overexpression attenuated increased RPA foci and RPA phosphorylation, inhibits DNA replication and cell growth, and ultimately lead to cell death. LMW-E overexpression facilitated DNA replication and cell proliferation with damaged DNA, resulting in multi-nuclei and micro-nuclei formation in daughter cells. Conclusion: LMW-E expression positively correlates with genomic instability in BC samples. LMW-E facilitates hMECs to by-pass replication stress induced tumor barrier and survive with damaged DNA, resulting in abnormal nuclei formation, an oncogenic phenotype. Citation Format: Mi Li, Kwang Huei Low, Tuyen Bui, Kelly K Hunt, Khandan Keyomarsi. Low molecular weight cyclin E facilitates replication stress tolerance in breast cancer development [abstract]. In: Proceedings of the 2019 San Antonio Breast Cancer Symposium; 2019 Dec 10-14; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2020;80(4 Suppl):Abstract nr P2-05-04.

A combination of thermochemical pretreatment and biological digestion technologies is usually required to overcome lignocellulosic recalcitrance and accomplish effective biomass deconstruction. This study is aimed at understanding switchgrass breakdown by hydrothermal, dilute acid, dilute alkali, and co-solvent enhanced lignocellulosic fractionation (CELF) pretreatments followed by application of traditional fungal enzymatic hydrolysis (EH) and Clostridium thermocellum consolidated bioprocessing (CBP) to the resulting solids. Unpretreated and pretreated switchgrass and their EH and CBP residues were characterized by a suite of analytical techniques to understand structural changes that occurred during deconstruction. CELF pretreated solids showed the highest accessibility and digestibility by both EH and CBP followed by dilute alkali and then dilute acid/hydrothermal pretreated solids. Lignin removal from biomass had a more positive impact on substrate accessibility and digestibility than did xylan removal, while xyloglucan removal by pretreatment appeared essential for cellulose digestion by fungal enzymes. The extent of CBP digestion of cellulose and non-cellulosic glycans was larger than that by EH. Unlike dilute alkali pretreatment, cellulose crystallinity increased for acid-based pretreatments in the following order: hydrothermal, dilute acid, and CELF. Acid-based pretreatments also substantially reduced cellulose degree of polymerization. All thermochemical and biological digestion approaches increased syringyl to guaiacyl lignin (S/G) ratio and reduced β-O-4 lignin interunit linkage and hydroxycinnamates content from levels in unpretreated switchgrass. The substantial increase in S/G ratio after hydrothermal and dilute alkali preatreatments suggested that high temperatures or alkali removed a large portion of G lignin from switchgrass.

Human embryo lung cellular protein interacting with severe acute respiratory syndrome-coronavirus nonstructural protein-10 (HEPIS) is expressed at varying levels in multiple organs and breast cancer cell lines. However, its expression and function in breast cancer cells has yet to be studied. Therefore, RNA in situ hybridization was used to detect the expression of HEPIS in breast cancer and cancer-adjacent normal breast tissue. HEPIS was expressed at lower levels in breast cancer compared with that in adjacent normal tissue. Subcellular localization of HEPIS was mainly found in the cytoplasm of HeLa cells. Cell Counting Kit-8 and 5-ethynyl-2'-deoxyuridine cell proliferation assays were used to investigate the role of HEPIS in cancer cell proliferation. Ectopic expression of HEPIS in MCF-7 cells was found to significantly inhibit cell proliferation. In contrast, knockdown of HEPIS by RNA interference exhibited the opposite effect. Furthermore, a dual-luciferase reporter assay was performed and HEPIS overexpression specifically inhibited the activity of the NF-κB reporter gene. Results of the gene chip assay revealed that 2,231 genes were differentially expressed in HEPIS-overexpressing cells. Results of the Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses indicated that these genes were enriched in the 'mitogen-activated protein kinase signaling pathway', 'JAK-STAT signaling pathway' and 'focal adhesion'. Reverse transcription-quantitative PCR was used to confirm the expression levels of the differentially expressed genes interleukin 2 receptor subunit α (IL2RA), interferon α and β receptor subunit 2 (IFNAR2) and IFα8 (IFNA8). In conclusion, the results of the present study indicated that HEPIS may function as a potential repressor of breast cancer.

Background: Egg cannibalism is common in nature. In China, Arma custos (Hemiptera: Asopinae) has been widely used as a natural enemy to control agricultural and forestry pests. A previous study showed that adult A. custos devour their eggs. However, no research has investigated the interaction between A. custos cannibalism and egg development. Clarifying the mechanisms involved in egg cannibalism by A. custos improves our understanding of the evolutionary relationships to enable more efficient mass rearing and biological control systems.Results: Virgin females showed a lower egg cannibalism inclination than gravid females. Both virgin and mated females showed a higher egg cannibalism inclination than virgin and mated males. The first and second instar nymphs did not devour eggs. The third, fourth, and fifth instar nymphs devoured eggs. Younger eggs were more readily eaten than older eggs. Neither A. custos nymphs nor female adults consumed all the available eggs, allowing an emergence ratio of >70%.Conclusion:Arma custos females exhibit a higher tendency for egg cannibalism than males. Egg cannibalism varies not only with the developmental stage of the eggs and nymphs but also with sex and reproductive status of A. custos females. These findings help us to better understand the evolutionary relationships in egg cannibalism by A. custos and contribute to the efficient mass rearing and realization of A. custos in biological control systems.

Osteosarcoma is one of the most common primary malignancies in bones and is characterized by high metastatic rates. Circulating tumor cells (CTCs) derived from solid tumors can give rise to metastatic lesions, increasing the risk of death in patients with cancer. Here, we used bioinformatics tools to compare the gene expression between CTCs and metastatic lesions in osteosarcoma to identify novel molecular mechanisms underlying osteosarcoma metastasis. We identified TRAIP as a key differentially expressed gene with prognostic significance in osteosarcoma. We demonstrated that TRAIP regulated the proliferation and invasion of osteosarcoma cells. In addition, we found that TRAIP promoted KANK1 polyubiquitination and subsequent degradation, downregulating IGFBP3 and activating the AKT pathway in osteosarcoma cells. These results support the critical role of the TRAIP/KANK1/IGFBP3/AKT signaling axis in osteosarcoma progression and suggest that TRAIP may represent a promising therapeutic target for osteosarcoma.

Simple Summary Triple-negative breast cancer (TNBC) is a subtype of invasive breast cancer with an aggressive phenotype that has decreased survival compared with other types of breast cancers, due in part to the lack of biomarker driven targeted therapies. Here, we show that breast cancer patients whose tumors show high levels of cyclin E expression have a higher prevalence of BRCA1/2 alterations and have the worst clinical outcomes. In vitro and in vivo studies revealed that combination therapies with poly (ADP-ribose) polymerase (PARP) and Wee1 kinase inhibitors in TNBC cells with either BRCA1 mutations or high levels of cyclin E results in synergistic cell death due to induction of replicative stress and downregulation of DNA repair. These studies suggest that by preselecting patients whose tumors have high cyclin E levels or harbor mutations in BRCA1, only those cases with the highest replicative stress properties will be subjected to combination treatment and likely result in synergistic activity of the two agents. Abstract The identification of biomarker-driven targeted therapies for patients with triple negative breast cancer (TNBC) remains a major clinical challenge, due to a lack of specific targets. Here, we show that cyclin E, a major regulator of G1 to S transition, is deregulated in TNBC and is associated with mutations in DNA repair genes (e.g., BRCA1/2). Breast cancers with high levels of cyclin E not only have a higher prevalence of BRCA1/2 mutations, but also are associated with the worst outcomes. Using several in vitro and in vivo model systems, we show that TNBCs that harbor either mutations in BRCA1/2 or overexpression of cyclin E are very sensitive to the growth inhibitory effects of AZD-1775 (Wee 1 kinase inhibitor) when used in combination with MK-4837 (PARP inhibitor). Combination treatment of TNBC cell lines with these two agents results in synergistic cell killing due to induction of replicative stress, downregulation of DNA repair and cytokinesis failure that results in increased apoptosis. These findings highlight the potential clinical application of using cyclin E and BRCA mutations as biomarkers to select only those patients with the highest replicative stress properties that may benefit from combination treatment with Wee 1 kinase and PARP inhibitors.

Background and objective Post-traumatic stress disorder (PTSD) is a chronic mental disorder caused by mental or psychological trauma after sudden events of a catastrophic or threatening nature. Synaptic plasticity is the core mechanism of PTSD and the main point of treatment of this disease. Methods Male Sprague Dawley rats were randomly divided into blank control (Ctrl), SPS (single-prolonged stress) model, SPS&S model (SPS and foot electric shock), SPS+EA (SPS plus electroacupuncture), and SPS&S+EA groups. Tranquilize Mind and Regulate Kidney (TMRK) electroacupuncture method was performed in each rat in the SPS+EA and SPS&S+EA groups, the treatment lasted for 20 minutes per day, simultaneously for 3 consecutive weeks. Behavioral evaluations, molecular tests, electron microscopy, electrophysiological testing were conducted following the treatment. Results First, electro-acupuncture can significantly improve the PTSD-like symptoms. Second, electro-acupuncture can up-regulate the long-term potentiation (LTP) in hippocampus, repair the synaptic morphology and improve BDNF levels in amygdala and hippocampus. Third, electroacupuncture can significantly up-regulate SYN, GAP43, and PSD95 protein levels and mRNA expression in amygdala and hippocampus. Conclusions The effect of TMRK electro-acupuncture method on the regression of fear memory of PTSD rats may be through its repair of synaptic plasticity in amygdala and hippocampus.

Notopterygium incisum C. C. Ting ex H. T. Chang (Apiaceae) is an endangered perennial herb in China. The lack of transcriptomic and genomic resources for N. incisum greatly hinders studies of its population genetics and conservation. In this study, we employed RNA-seq technology to characterize transcriptomes for the flowers, leaves, and stems of this endangered herb. A total of 56 million clean reads were assembled into 120,716 unigenes with an N50 length of 850 bp. Among these unigenes, 70,245 (58.19%) were successfully annotated and 65,965 (54.64%) were identified as coding sequences based on their similarities with sequences in public databases. We identified 21 unigenes that had significant relationships with cold tolerance in N. incisum according to gene ontology (GO) annotation analysis. In addition, 13,149 simple sequence repeats (SSRs) and 85,681 single nucleotide polymorphisms were detected as potential molecular genetic markers. Ninety-six primer pairs of SSRs were randomly selected to validate their amplification efficiency and polymorphism. Nineteen SSR loci exhibited polymorphism in three natural populations of N. incisum. These results provide valuable resources to facilitate future functional genomics and conservation genetics studies of N. incisum. Keywords: Endangered species, EST-SSR marker, Notopterygium incisum, Single nucleotide polymorphism, Transcriptome

Cell wall recalcitrance is the major obstacle for plant biomass conversion to biofuels. In this study, we functionally characterized a serine hydroxymethyltransferase (SHMT) from Populus and evaluated its potential for developing lignocellulosic feedstocks. SHMT is an enzyme that plays an important role in cellular one-carbon pathways. However, little is known about its function in plant cell wall-related processes. Among nine SHMT genes in the Populus genome, PtSHMT2 was highly expressed in the developing xylem and was co-expressed with secondary cell wall biosynthetic genes. In Populus transgenic plants overexpressing PdSHMT2, the biomass yield and sugar (glucose and xylose) release were increased whereas the lignin content was decreased. Transcriptomics and metabolomics analyses revealed that genes and metabolites related to secondary cell wall biosynthesis were affected by PdSHMT2 overexpression. Based on the transcription factor binding sites of differentially expressed genes in PdSHMT2 overexpression lines, a total of 27 hub transcription factors were identified. We proposed a regulatory model of action of PdSHMT2 with transcriptional master switches of secondary cell wall biosynthesis. Collectively, these results suggest that PdSHMT2 is a promising candidate for genetic engineering to improve feedstock characteristics to enhance biofuel conversion and reduce the cost of lignocellulosic biofuel production.

With the rapid development of information technology and biomedical engineering, people can get more and more information. At the same time, they begin to study how to apply the advanced technology in biomedical information. The main research of this paper is to optimize the machine learning method by particle swarm optimization (PSO) and apply it in the classification of biomedical data. In order to improve the performance of the classification model, we compared the different inertia weight strategies and mutation strategies and their combinations with PSO, and obtained the best inertia weight strategy without mutation, the best mutation strategy without inertia weight and the best combination of the two. Then, we used the three PSO algorithms to optimize the parameters of support vector machine in the classification of biomedical data. We found that the PSO algorithm with the combination of inertia weight and mutation strategy and the inertia weight strategy that we proposed could improve the classification accuracy. This study has an important reference value for the prediction of clinical diseases.

Background: Ewing sarcoma (ES) is one of the most common types of round cell mesenchymal neoplasms related to children and young adults. It is characterized by chromosomal translocations. However, that does not completely explain the ES proliferation and development. Methods: Several public data series (GSE45544, GSE68591, and GSE68776) were used to identify differentially expressed genes (DEGs) between ES and normal tissue. Furthermore, functional enrichment analysis and protein-protein interaction network of DEGs were performed. The regulatory network of DEGs and hub genes, survival analysis of hub genes was visualized. In addition, functional predictions of the candidate gene were analyzed. Results: A total of 142 DEGs 10 hub genes were identified. While out of them, only one candidate gene FGF7 was found to be suitable as a candidate gene. Conclusions: In conclusion, the DEGs, hub genes, and their regulatory molecules screened out in this research could contribute to comprehend the mechanisms in ES and provide potential research molecular for further study.

MITA (also known as STING) is an ER-located adaptor protein, which mediates DNA-triggered innate immune response and is critically involved in autoimmune diseases and tumorigenesis. MITA is regulated by post-translational modifications, but how post-transcriptional mechanisms are involved in the regulation of MITA is still largely unknown. Here, we identified the RNA-binding protein LUC7L2 as a negative regulator of DNA virus-triggered innate immune response. LUC7L2-deficient mice exhibited resistance to lethal herpes simplex virus 1 (HSV-1) infection and reduced HSV-1 loads in the brain. Mechanistically, LUC7L2 directly bound to intron 3 of MITA precursor messenger RNA, inhibited its splicing and promoted its nonsense-mediated decay, leading to its downregulation at protein level. LUC7L2-deficient cells had markedly increased MITA level, leading to heightened innate antiviral response. Finally, LUC7L2 was induced following HSV-1 infection. Our findings reveal a feedback negative post-transcriptional regulatory mechanism for regulation of MITA-mediated innate immune response to viral and aberrant cellular DNA.

Lung cancer is a very aggressive cancer characterized with molecular heterogeneities in different subtypes, including lung adenocarcinoma and lung squamous cell carcinoma. However, few related molecular signatures have been established for the treatment of lung cancer subtypes. Polo‐like kinase (PLK) family is a crucial regulator during cell division. Aberrant genetic and epigenetic alteration of PLK members plays a controversial role among different cancers. In this study, we performed an analysis of transcriptional and protein expression to identify overexpressed PLK1/4 and under‐expressed PLK2/3 in lung cancer subtypes. We then analysed biological function of PLKs and related genes. Besides, we estimated a correlation of PLKs with patient's genders and TP53 mutation in lung cancer. Higher PLK1/4 expression was significantly associated with male patient and TP53 mutant status, separately. Moreover, we carried out a methylation profile analysis including methylation level, DNA methyltransferases correlation and survival analysis of global methylation. Global methylation survival analysis showed that prognostic value of PLK1/2/4 methylation remained the same significant trend between two lung cancer subtypes, whereas prognostic value of PLK3 methylation lacked consistency. Taken together, these results provided instructive insights into a comprehensive evaluation for advanced therapeutic strategy based on epigenetic evidences.

Exosomes have emerged as a new drug delivery system. In particular, exosomes derived from mesenchymal stem cells (MSCs) have been extensively studied because of their tumor-homing ability and yield advantages. Considering that MSC-derived exosomes are a double-edged sword in the development, metastasis, and invasion of tumors, engineered exosomes have broad potential use. In this review, we focused on the latest development in the treatment of tumors using engineered and nonengineered MSC-derived exosomes (MSC-EXs). Nonengineered MSC-EXs exert an antitumor effect on several well-studied tumors by affecting tumor growth, angiogenesis, metastasis, and invasion. Furthermore, engineered exosomes have promising research prospects as drug-carrying tools for the transport of miRNAs, small-molecule drugs, and proteins. Although exosomes lack uniform standards in terms of definition, separation, and purification, they still have great research value because of their unique advantages, such as high biocompatibility and low toxicity. Future studies on MSC-EXs should elucidate the mechanisms underlying their anticancer effect and the safety of their application.

Sulfonamides (SAs) are widespread in soils, and their dissipation behavior is important for their fate, risk assessment, and pollution control. In this work, we investigated the dissipation behavior of different SAs in a soil under aerobic condition, focusing on revealing the relationship between overall dissipation (without sterilization and in dark) and individual abiotic (sorption, hydrolysis)/biotic (with sterilization and in dark) factors and taxonomy/function of microbiomes. The results showed that dissipation of all SAs in the soil followed the pseudo-first-order kinetic model with dissipation time at 50% removal (DT50) of 2.16-15.27 days. Based on, experimentally, abiotic/biotic processes and, theoretically, partial least-squares modeling, a relationship between overall dissipation and individual abiotic/biotic factors was developed with microbial degradation as the dominant contributor. Metagenomic analysis showed that taxonomic genera like Bradyrhizobium/Sphingomonas/Methyloferula and functions like CAZy family GT51/GH23/GT2, eggNOG category S, KEGG pathway ko02024/ko02010, and KEGG ortholog K01999/K03088 are putatively involved in SA microbial degradation in soil. Spearman correlation suggests abundant genera being multifunctional. This study provides some new insights into SA dissipation and can be applied to other antibiotics/soils in the future.

The predatory insect Arma chinensis (Hemiptera: Pentatomidae) is widely distributed in China, where it is also used to control many agricultural and forest pests. The chemosensory genes expressed in its antennae play crucial roles in food-seeking and mating behaviors. To better understand the olfaction of A. chinensis antennae, we identified the genes related to food-seeking and mating. Sequencing of the antennal transcriptomes of full and hungry male and female A. chinensis revealed 38 odorant-binding proteins (OBPs), 1 chemosensory protein (CSP), 1 Niemann-Pick C2 protein (NPC2), 3 odorant receptors (ORs), 12 ionotropic receptors (IRs), 2 gustatory receptors (GRs), and 3 sensory neuron membrane proteins (SNMPs). These results were used to construct phylogenetic trees. A quantitative real-time PCR (qRT-PCR) analysis showed that the relative transcript levels of AchiGR1, AchiGR2, and AchiOBP28 were higher in female than in male antennae in both full and hungry insects, but that the expression of AchiOBP13 and AchiOBP16 was higher only in full A. chinensis females. Thus, the latter genes may encode proteins involved in oviposition selection behavior. AchiGRs (1 and 2), AchiIR6, and AchiOBPs (6-8, 12, 20-22, 28, and 34) were highly expressed only in the antennae of full males, indicating the participation of these genes in mate-searching or male pheromone recognition. The expression of AchiOBP31 in the antennae of starved males, AchiOBPs (15, 18, and 29) in the antennae of starved females, and AchiOBPs (3, 4, and 24) in the antennae of starved males and females suggested that these genes encode food-seeking functions. Our identification of chemosensory genes in A. chinensis antennae and their differential expression in full and hungry insects provides the basis for further functional studies on the chemoreception system of A. chinensis and the sex hormones of predatory insects.

Osteosarcoma is one of the most common types of bone sarcoma with a poor prognosis. However, genes involved in the mineral metabolism in the microenvironment of the bone affected by osteosarcoma are, to date, largely unknown. A public data series (GSE114237) was used to identify differentially expressed genes (DEGs) between osteosarcoma cells adhering to demineralized osseous surfaces and mineralized osseous surfaces. Functional enrichment analysis of DEGs and hub genes, protein-protein interaction network of DEGs and regulatory network (miRNA-mRNA network and transcription factor (TF)-mRNA network), survival analysis of hub genes was visualized. The prognostic hub genes were considered as candidate genes and their functional predictions were analyzed. A total of 207 DEGs were mainly enriched in extracellular space and thirteen hub genes were mainly enriched in the function of epithelial to mesenchymal transition. However, out of these, only one candidate gene was found to be suitable as a candidate gene. Besides that, 297 miRNAs and 349 TFs interacting with the hub genes were screened. In conclusion, the DEGs, hub genes, miRNAs and TFs screened out in this research could contribute to comprehend the latent mechanisms in osteosarcoma affected by matrix mineral and provide potential research molecular for further study.

Except remdesivir, no specific antivirals for SARS-CoV-2 infection are currently available. Here, we characterize two small-molecule-compounds, named GRL-1720 and 5h, containing an indoline and indole moiety, respectively, which target the SARS-CoV-2 main protease (Mpro). We use VeroE6 cell-based assays with RNA-qPCR, cytopathic assays, and immunocytochemistry and show both compounds to block the infectivity of SARS-CoV-2 with EC50 values of 15 ± 4 and 4.2 ± 0.7 μM for GRL-1720 and 5h, respectively. Remdesivir permitted viral breakthrough at high concentrations; however, compound 5h completely blocks SARS-CoV-2 infection in vitro without viral breakthrough or detectable cytotoxicity. Combination of 5h and remdesivir exhibits synergism against SARS-CoV-2. Additional X-ray structural analysis show that 5h forms a covalent bond with Mpro and makes polar interactions with multiple active site amino acid residues. The present data suggest that 5h might serve as a lead Mpro inhibitor for the development of therapeutics for SARS-CoV-2 infection., Here, using in vitro assays and structural analysis, the authors characterize the anti-SARS-CoV-2 properties of two small molcules, showing these to bind and target the virus main protease (Mpro), and to exhibit a synergistic antiviral effect when combined with remdesivir in vitro.

Background Zinc oxide nanoparticle (nZnO) and chemicals with different mode of action (MOA, i.e., narcotic and reactive) were frequently detected in the Yangtze River. Organisms are typically exposed to mixtures of nZnO and other chemicals rather than individual nZnO. Toxicity of nZnO is caused by the dissolution of Zn2+, which has been proved in the field of single toxicity. However, it is still unclear whether the released Zn2+ plays a critical role in the nZnO toxicity of nZnO–chemicals mixtures. In the present study, the binary mixture toxicity of nZnO/Zn2+ and chemicals with different MOA was investigated in acute (15 min) and chronic (12 h) toxicity test upon Vibrio fischeri (V. fischeri). The joint effects of nZnO and tested chemicals were explored. Moreover, two classic models, concentration addition (CA) and independent action (IA) were applied to predict the toxicity of mixtures. Results The difference of toxicity unit (TU) values between the mixtures of Zn2+–chemicals with those of nZnO–chemicals was not significant (P > 0.05), not only in acute toxicity test but also in chronic toxicity test. The antagonistic or additive effects for nZnO-chemicals can be observed in most mixtures, with the TU values ranging from 0.75 to 1.77 and 0.47 to 2.45 in acute toxicity test and chronic test, respectively. We also observed that the prediction accuracy of CA and IA models was not very well in the mixtures where the difference between the toxicity ratios of the components was small (less than about 10), with the mean absolute percentage error (MAPE) values ranging from 0.14 to 0.67 for CA model and 0.17–0.51 for IA model, respectively. Conclusion We found that the dissolved Zn2+ mainly accounted for the nZnO toxicity in the mixtures of nZnO–chemicals, and the joint effects of these mixtures were mostly antagonism and additivity. CA and IA models were unsuitable for predicting the mixture toxicity of nZnO–chemicals at their equitoxic ratios.

Post-traumatic stress disorder (PTSD) is a mental health condition triggered by a terrifying event, causing flashbacks, nightmares and severe anxiety. It develops in individuals who have experienced a shocking, scary, or dangerous event. Electroacupuncture is reported to be effective for the treatment of PTSD. The present study was carried out to investigate the protective effect of electroacupuncture in a rat model of PTSD, and the mechanism involved. Specific-pathogen-free male Sprague Dawley rats (n = 30) weighing 180 – 220 g (mean weight = 200 ± 20 g) were randomly assigned to three groups of ten rats each: control group, single-prolonged stress (SPS) group, and treatment group. The treatment group rats received electroacupuncture. Changes in PTSD-like behavior were assessed using locomotor activity, elevated plus-maze (EPM) and fear conditioning tests. The mRNA and protein expressions of brain-derived neurotrophic factor (BDNF) and tropomyosin receptor kinase B (TrkB) were determined using real-time quantitative polymerase chain reaction (qRT-PCR) and Western blotting, respectively. Co-immunoprecipitation (Co-IP) was used to measure BDNF and TrkB binding interaction, while chromatin immunoprecipitation (ChIP) was used to evaluate the binding between cyclic adenosine monophosphate (cAMP) response element-binding protein (CREB) and its target genes. Electroacupuncture significantly increased locomotor activity and exploratory behavior, but significantly reduced general fear and anxiety in SPS rats (p < 0.05). It also significantly upregulated the mRNA and protein expressions of BDNF and TrkB, and increased the binding of BDNF to its receptor TrkB (p < 0.05). Electroacupuncture significantly increased the binding of CREB to BDNF promoter region (p < 0.05). Electroacupuncture ameliorates PTSD in rats via a mechanism involving the BDNF-TrkB signaling pathway.

Summary Prefoldin (PFD) is a group II chaperonin that is ubiquitously present in the eukaryotic kingdom. Six subunits (PFD1‐6) form a jellyfish‐like heterohexameric PFD complex and function in protein folding and cytoskeleton organization. However, little is known about its function in plant cell wall‐related processes. Here, we report the functional characterization of a PFD gene from Populus deltoides, designated as PdPFD2.2. There are two copies of PFD2 in Populus, and PdPFD2.2 was ubiquitously expressed with high transcript abundance in the cambial region. PdPFD2.2 can physically interact with DELLA protein RGA1_8g, and its subcellular localization is affected by the interaction. In P. deltoides transgenic plants overexpressing PdPFD2.2, the lignin syringyl/guaiacyl ratio was increased, but cellulose content and crystallinity index were unchanged. In addition, the total released sugar (glucose and xylose) amounts were increased by 7.6% and 6.1%, respectively, in two transgenic lines. Transcriptomic and metabolomic analyses revealed that secondary metabolic pathways, including lignin and flavonoid biosynthesis, were affected by overexpressing PdPFD2.2. A total of eight hub transcription factors (TFs) were identified based on TF binding sites of differentially expressed genes in Populus transgenic plants overexpressing PdPFD2.2. In addition, several known cell wall‐related TFs, such as MYB3, MYB4, MYB7, TT8 and XND1, were affected by overexpression of PdPFD2.2. These results suggest that overexpression of PdPFD2.2 can reduce biomass recalcitrance and PdPFD2.2 is a promising target for genetic engineering to improve feedstock characteristics to enhance biofuel conversion and reduce the cost of lignocellulosic biofuel production.

A novel Gram-stain-negative, catalase- and oxidase-positive, motile, short rod-shaped bacterium designated BGMRC 6574T was isolated from stems of Aegiceras corniculatum collected from Hainan province, China. The strain grew at 25–37 °C (optimal at 28 °C), pH 5.0–10.0 (pH 7.0), and 3–8% (w/v) NaCl (3%). Based on the 16S rRNA phylogenetic analysis, the strain was closely related to Pararhizobium haloflavum MCCC 1K03228T (96.45% sequence similarity). The novel strain showed an average nucleotide identity value and a digital DNA–DNA hybridization of 72.62 and 27.1%, respectively, to P. haloflavum MCCC 1K03228T based on draft genome sequences. The G+C content of the genomic DNA was 64.7 mol%. The major respiratory quinone was Q-10. The strain possessed genes putatively encoding choline uptake and conversion to betaine gene clusters. The extract significantly delayed the lifespan of Caenorhabditis elegans compared to the control (P

Delignification of Broussonetia papyrifera during ethanol organosolv pretreatment was studied. Milled wood lignin (MWL), the ethanol organosolv lignin recovered at 5 min (PL5), 10 min (PL10), and 60 min (PL60) during the pretreatment, and the residual lignin (RL60) after the pretreatment were characterized and compared. GPC results demonstrated that the weight-average molecular weights (Mw) of the ethanol organosolv lignin fractions increased as the pretreatment time extended from 5 to 60 min. 31P NMR analysis revealed that the content of aliphatic OH decreased in the following order: RL60 < PL60 < PL5 < PL10 < MWL. HSQC NMR results suggested that the contents of syringyl, guaiacyl, p-hydroxybenzoate unit, phenylcoumaran, and resinol in the solubilized lignin fractions were significantly changed during the pretreatment. In addition, a significant portion of β-O-4 interunit linkages of lignin was cleaved during the pretreatment, leading to the formation of stilbene structures in lignin samples such as PL60.

Emerging studies show that dysregulated miRNAs are implicated in tumorigenesis and progression of various cancers. MiRNA-320c, an important member of miRNA-320 family, was characterized as a new candidate miRNA that suppressed the development of colorectal cancer and bladder cancer. However, the function of miRNA-320c in human glioma remained unclear. Here, we found that miRNA-320c was significantly down-regulated in glioma tissues in contrast with normal brain tissues, being tightly related to clinical stage of glioma by qRT-PCR. Moreover, Kaplan-Meier analysis demonstrated that patients with low miRNA-320c expression had a shorter survival. Multivariate Cox regression analysis indicated that miRNA-320c could serve as an independent poor prognostic factor for patients with glioma. Functionally, overexpression of miRNA-320c could dramatically inhibit glioma cell proliferation, migration and invasion, as well as promote apoptosis. Further analysis indicated that overexpression of miRNA-320c dramatically led to the G0/G1 phase arrest and correspondingly decreased the percentage of S phase cells by suppressing the expression of G1/S transition key regulators, such as Cyclin D1 and CDK6. Additionally, up-regulation of miRNA-320c could significantly impair migration and invasion of glioma cells via reducing the expression of MMP2, MMP9, N-cadherin and Integrin β1. Collectively, our data revealed that miRNA-320c played a crucial role in the carcinoma processes of glioma and might serve as a new prognosis biomarker and therapeutic target of glioma.

Endophytic fungi strains (n = 81) were isolated from the leaves, barks, and fruits of Camellia oleifera from Hunan province (China) to delineate their species composition and potential as biological control agents of C. oleifera anthracnose. The fungi were identified by morphological and phylogenetic analyses. Fungal colonization rates of the leaves, barks, and fruits were 58.02, 27.16, and 14.81%, respectively. The isolates were identified as 14 genera, belonging to two subdivisions, Deuteromycotina and Ascomycotina; 87.65% of all isolates belonged to Deuteromycotina. The dominant species, occurring with a high relative frequency, were Pestalotiopsis sp. (14.81%), Penicillium sp. (14.81%), and Fusarium sp. (12.35%). The Simpson’s and Shannon’s diversity indices revealed the highest species diversity in the leaves, followed by the barks and fruits. The similarity index for the leaves versus barks comparison was the highest, indicating that the number of endophytic fungal species shared by the leaves and barks was higher than barks and fruits or leaves and fruits. Based on the results of dual culture experiments, only five strains exhibited antifungal activity against C. oleifera anthracnose pathogen, with isolate ty-64 (Oidium sp.) generating the broadest inhibition zones. Our results indicate that the endophytes associated with C. oleifera could be employed as natural agents controlling C. oleifera anthracnose.

Thirteen compounds were isolated from the lipid-soluble extracts of Illicium ternstroemioides A. C. Smith, including eleven previously undescribed prenylated C6–C3 compounds, a previously undescribed prenylated C6–C3 derivative-abscisic acid ester hybrid, and a known compound (4S)-illicinone I. Their structures and configurations were mainly elucidated by spectroscopic analyses, CD experiments and X-ray crystallography. (2S,4R,11S)-4-O-methyl-12-chloroillifunone C, (2S,4R,11R)-2,3-dihydro-4-O-methyl illioliganfunone D, and illiternfunol A were found to exhibit weak activity against Coxsackievirus B3, with IC50 values ranging from 27.8 to 33.3 μM. Illiternone B exhibited more potent activities against Coxsackievirus B3 and influenza virus A than did its geometric isomer illiternone A, with IC50 values of 7.7 μM and 2.5 μM, respectively. None of these compounds displayed cytotoxic activities.

A series of new 6-aryl-1-(3,4,5-trimethoxyphenyl)-1H-benzo[d]imidazoles as tubulin polymerization inhibitors targeting the colchicine-binding site were designed to restrict bioactive configuration of (Z,E)-vinylogous CA-4. All of the target compounds were synthesized and then evaluated for their in vitro antiproliferative activities. Among them, 2a exhibited the most potent activities against three cancer cell lines with IC50 values in the range of 0.037–0.20 μM. Further mechanism studies revealed that 2a inhibited tubulin polymerization, disrupted cell microtubule networks, arrested the cell cycle at G2/M phase, induced apoptosis and hindered cancer cell migration. Moreover, 2a displayed significant in vivo antitumor efficacy in 4T1-xenograft mice model with tumor growth inhibition rate of 52% at the dose of 2.5 mg/kg. Colchicine competition assay and the docking model of 2a in complex with tubulin showed that 2a acted at the colchicine-binding site.

Two series of 2,7-diaryl-pyrazolo[1,5-a]pyrimidines as tubulin polymerization inhibitors were designed to restrict bioactive configuration of (E,Z)-vinylogous CA-4. All of the target compounds were synthesized and then evaluated for their in vitro antiproliferative activities against three cancer cell lines (MCF-7, SGC-7901 and A549). Among them, 6d exhibited the most potent antiproliferative activity against the MCF-7 with IC50 value of 0.047 μM. Moreover, 6d significantly inhibited tubulin polymerization, disrupted microtubule networks, arrested cell cycle at G2/M phase, induced apoptosis and hindered cancer cell migration. Colchicine competition assay and molecular docking studies suggested that 6d could interact with tubulin by binding to the colchicine site.

Lignin S/G ratio has been investigated as an important factor in biomass recalcitrance to bioethanol production. Because of the complexity and variety of biomass, recalcitrance was also reportedly influenced by several other factors, such as total lignin content, degree of cellulose polymerization, etc. In addition, the effect of S/G ratio on biomass conversion is not uniform across plant species. Herein, 11 Populus trichocarpa natural variants grown under the same conditions with similar total lignin content were selected to minimize the effects of other factors. The lignin S/G ratio of the selected P. trichocarpa natural variants showed negative correlations with p-hydroxybenzoate (PB) and β–5 linkage contents, while it had positive ones with β-O-4 linkage, lignin molecular weight, and ethanol production. This study showed the importance of lignin S/G ratio as an independent recalcitrance factor that may aid future energy crop engineering and biomass conversion strategies.

Broussonetia papyrifera, known as paper mulberry, is a potential feed stock for bioethanol production because of its cellulose-rich composition. Lignin in dilute acid pretreated Broussonetia papyrifera was fractionated to three different fractions, and their physiochemical properties were determined by FT-IR, GPC and NMR analyses. Different structural characteristics were observed from each lignin fraction. Cellobiohydrolases I (CBH) adsorption to each lignin was understood by the lignin properties. The results showed that aliphatic hydroxyl groups in lignin showed positive correlations with the maximum binding ability of CBH onto lignin samples. Also, the contents of phenolic compounds such as p-hydroxyphenyl benzoate (PB), syringyl (S) and guaiacyl (G) units in the lignin influenced their CBH binding.

The climate oscillations of the quaternary periods have profoundly affected the geographic distributions of current species. Acer davidii is a deciduous forest tree species mainly distributed in East Asia and China, playing a dominant role in the local forest ecosystem. In order to study the potential changes of geographic distributions of A. davidii in climate fluctuations, we collected the relate geographical distribution data and six climatic variables, using maximum entropy modelling to determine the species distribution. The results showed that the Areas Under Curve (AUC) values of the working characteristic curves of the subjects in the five historical periods were all greater than 0.93, suggesting that the results of maximum entropy modelling were accurate. The simulation of species distribution showed that the suitable area of A. davidii was mainly concentrated in central and northern China in contemporary times. From the Last Interglacial Age (LIG) to the Last Glacial Maximum (LGM), and then to the future (2050, 2070), the distribution area of this species experienced a decrease (LGM~Current, the high adaptability areas of central China became moderate) then an increase (Current~2050, the adaptation areas expanded to South Asia, Southeast Asia, and Siberia), and finally decreased (2050~2070, the suitable areas of South Asia, Southeast Asia, and Siberia shrank returning to China at latitude 25 °N). Compared to the LGM, the area of contemporary suitable area increased. Interestingly, the area of suitable growth range under future climatic conditions (2050) increased by half than before, and the suitable distribution area moved from Midwest China to Northeast China. This study on the change of species distribution can provide a typical case for the model study on the response of plants to climate change in the north temperate and subtropical zones of East Asia. Meanwhile, it can also give a basis for planting planning, species protection, and management.

Lignin offers structural support and protection for plant cell walls; however, it also contributes to biomass recalcitrance and the costs of biofuel production via the biological pathway. Organosolv and ammonia pretreatments have been developed to reduce biomass recalcitrance and improve sugar release performance during enzymatic hydrolysis. It is believed that lignin properties are related to its inhibition on enzymatic hydrolysis; therefore, understanding the characteristics of lignin is a key for effective biomass conversion to biofuels. In this study, an organosolv pretreatment using 60% ethanol with 1.25% H2SO4 significantly deconstructed poplar lignin and reduced its molecular weights due to the cleavage of lignin inter-unit linkages. The organosolv pretreatment increased the contents of phenolic OH units and the lignin residue showed a high cellulase maximum adsorption capacity. Ammonia pretreatment with 5% ammonium hydroxide was not as effective as organosolv pretreatment on lignin deconstruction. Organosolv lignin residue had lower lignin S/G ratio than the untreated one. Compared to the organosolv lignin residue and untreated lignin, ammonia lignin residue had a higher cellulase adsorption affinity. In addition, the effects of lignin on cellulose hydrolysis was investigated and the results suggested that the presence of lignin with cellulose substrates reduced cellulose hydrolysis, and its inhibitory effect was primarily determined by the lignin properties after each pretreatment. The organosolv pretreatment resulted in a slightly lower cellulase binding strength (249.7 mL g−1) on poplar lignin than that on untreated samples (261.1 mL g−1), while ammonia lignin residue showed a higher cellulase binding strength (402.8 mL g−1) and had more significant inhibition effect on cellulose hydrolysis. These results demonstrated that the binding strength significantly affected the lignin-derived inhibition on enzymatic hydrolysis of cellulose in the cellulose-lignin mixtures.

Morchella eohespera Beug, Voitk & O’Donnell is a typical black morel species. In this study, using the Nanopore sequencing platform, we characterized its whole mitochondrial (mt) genome sequence. Mt genome of M. eohespera is composed of circular DNA molecules of 243,963 bp, which encoded 102 protein-coding genes (PCGs), two ribosomal RNA genes (rRNA), and 31 transfer RNA (tRNA) genes. The base composition of M. eohespera mitogenome is as follows: A (30.40%), T (29.30%), G (20.8%), and C (19.5%). The phylogenetic analysis suggested that M. eohespera was closely related to the congeneric M. importuna.

Zanthoxylum armatum DC. (Rutaceae) is a shrub and/or tree species with the important medicinal and economic values. In this study, the plastid genome of Z. armatum was characterized by Illumina Hiseq 2500 sequencing platform. In total, the plastid genome is 158,557 bp in length, and comprises a large single copy region of 85,752 bp, a small single copy region of 17,605 bp, and two inverted repeat regions of 27,600 bp. The complete plastid genome contains 87 protein-coding genes, 37 tRNA genes, and 8 rRNA genes. Phylogenetic analysis suggested that Z. armatum and the congeneric Z. simulans clustered into an evolutionary clade with the high support.

Forsythia mira M. C. Chang (Oleaceae) is an endemic medicinal shrub in China. In this study, we first characterized its whole plastid genome sequence using the Illumina sequencing platform. The pla...

Anemone taipaiensis W. T. Wang is an endemic herb species in Shaanxi province (China). Here, we first characterized its whole plastid genome via pair-end sequencing method. The whole chloroplast genome was 156,659 bp in size, including a large single-copy (LSC) region of 78,439 bp, a small single-copy (SSC) region of 16,178 bp, and two repeat regions (IRs) of 31,021 bp. A total of 135 genes, including 91 protein-coding genes, 36 tRNA, and 8 rRNA genes were identified in A. taipaiensis. The phylogenetic analysis showed that A. taipaiensis have a close relationship with congeneric species A. trullifolia.

Background Osteosarcoma is a primary malignant tumor originating from mesenchymal tissue, with a poor distant metastasis prognosis. The molecular mechanisms of osteosarcoma metastasis are extremely complicated. Methods A public data series (GSE21257) was used to identify differentially expressed genes (DEGs) in osteosarcoma patients that did, or did not, develop metastases. Functional enrichment analysis, a protein-protein interaction network, and survival analysis of DEGs were performed. DEGs with a prognostic value were considered as candidate genes and their functional predictions, different expression in normal and malignant tissues, and immune infiltration were analyzed. Results The DEGs were mainly enriched in the immune response. Three candidate genes (ALOX5AP, CD74, and FCGR2A) were found, all of which were expressed at higher levels in lungs and lymph nodes than in matched cancer tissues and were probably expressed in the microenvironment. Conclusions Candidate genes can help us understand the molecular mechanisms underlying osteosarcoma metastasis and provide targets for future research.

Background The empiric therapies for H. pylori infection in clinical guidelines have been widely used. However, the cure rate is decreasing due to the increase of antibiotic resistance. The importance of antimicrobial susceptibility test (AST) has been documented in many consensuses. However, the effect of tailored therapy remains controversial because the AST is rarely offered in most areas. We compared the evolution of treatment efficacy among tailored therapy and some recommended empiric therapies through a trend survey from 1999 to 2018. Methods This retrospective survey was performed at 2 medical centers and 3 community hospitals in Taiwan. A total of 16,370 treatment naive or failure patients were recruited. Successful H. pylori eradication was defined as a negative 13C-UBT. The empiric first-line regimens include tailored therapy, clarithromycin-containing triple therapy (CLA-TT), sequential therapy (ST), bismuth-containing quadruple therapy (BQT), and high-dose dual therapy (HDDT). The empiric rescue regimens include tailored therapy, levofloxacin-containing triple therapy (LEV-TT), BQT, and HDDT. We divided the 20 years of follow-up time into 4-year periods for evaluating the trend of treatment efficacies. The E-test was performed to evaluate H. pylori resistance. For the tailored therapy, CLA-TT, LEV-TT, BQT, or HDDT was chosen according to the resistance pattern of each patient. Results The efficacies of tailored therapy, BQT, and HDDT maintain a stable and high efficacy in both first-line and rescue treatment during the study period. However, the efficacies of CLA-TT, ST, and LEV-TT are decreasing year by year. The eradication rate of tailored therapy is significantly higher than that of CLA-TT, ST, LEV-TT, and BQT in recent 4-year period. However, there is no significant difference between the efficacy of tailored therapy and HDDT. The prevalence of H. pylori resistance to CLA and LEV increased gradually. In contrast, the resistance rate to amoxicillin and tetracycline remained low. Conclusions Over the past 20 years, we found that the efficacy of tailored therapy remains relatively stable. Of the recommended empiric therapies, HDDT and BQT have stable therapeutic efficacies and are a good choice of empiric treatment currently.