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Plant-derived natural active products have attracted increasing attention for use in flavors and perfumes. These compounds also have applications in insect pest control because of their environment-friendly properties. Holy basil (Ocimum sanctum), a famous herb used in Ayurveda in India, is a natural source of medical healing agents and insecticidal repellents. Despite the available genomic sequences and genome-wide bioinformatic analysis of terpene synthase genes, the functionality of the sesquiterpene genes involved in the unique fragrance and insecticidal activities of Holy basil are largely unknown. In this study, we systematically screened the sesquiterpenoid biosynthesis genes in this plant using a precursor-providing yeast system. The enzymes that synthesize β-caryophyllene and its close isomer α-humulene were successfully identified. The enzymatic product of OsaTPS07 was characterized by in vivo mining, in vitro reaction, and NMR detection. This product was revealed as (-)-eremophilene. We created a mutant yeast strain that can achieve a high-yield titer by adjusting the gene copy number and FPP precursor enhancement. An optimized two-stage fed-batch fermentation method achieved high biosynthetic capacity, with a titer of 34.6 g/L cyclic sesquiterpene bioproduction in a 15-L bioreactor. Further insect-repelling assays demonstrated that (-)-eremophilene repelled the insect pest, fall leafworm, suggesting the potential of (-)-eremophilene as an alternative to synthetic chemicals for agricultural pest control. This study highlights the potential of our microbial platform for the bulk mining of plant-derived ingredients and provides an impressive cornerstone for their industrial utilization.

Three new isopimaric acid diterpenoids, 6-oxoisopimaric acid (1), 6α-hydroxyisopimaric acid (2), and isopimara-7,9(11),15-trien-18-oic acid (4), together with two known isopimaric acid diterpenoids, isopimaric acid (3), and 8(14),15-isopimaradien-18-oic acid (5), were isolated from the bark of Cryptomeria japonica D. Don. Their structures were determined by analysis of spectroscopic data and comparison with the spectral data of known analogues. At the concentration of 50 μM, compounds 1–5 inhibited xanthine oxidase activity by 17.3, 16.5, 2.6, 30.5, and 24.5 %, respectively.

The objective of this study was to explore the effects and underlying intervention mechanisms of Angelica water extract (AWE) on abnormal uterine bleeding (AUB) based on serum metabolomics. Firstly, the concentration of main active substances in AWE was determined and the chemical components were identified by UPLC‐Q‐Exactive Orbitrap‐MS/MS. A drug‐induced abortion model was established by mifepristone and misoprostol. After administration AWE (2.16 g/kg) for 7 days, the coagulation function, serum hormone levels, H&E staining, and immunohistochemistry observation of uterus were detected. In addition, serum metabolites profiles were performed on ultra‐performance liquid chromatography–quadrupole–time‐of‐flight mass spectrometry (UPLC‐Q‐TOF‐MS). The contents of ferulic acid, senkyunolide A, and ligustilide in AWE were 0.7276, 0.0868, and 1.9908 mg/g, respectively. Twenty‐six compounds were identified in AWE. It was found that AWE was effective in regulation of coagulation function and promoting endometrial recovery. Meanwhile, the levels of E2, Pg, and HCG and the expression of ERα, Erβ, and PR were down‐regulated in AUB model and up‐regulated by the treatment of AWE. Twenty‐one potential biomarkers were eventually identified by multivariate statistical analysis. Study indicated that glycerophospholipid, sphingolipid, amino acids, retinol metabolism and primary bile acid biosynthesis were the main related metabolic pathways involved for the treatment of AUB by AWE. The results showed that AWE has potential therapeutic effect on AUB by altering the metabolic aberrations., Angelica water extract could reduce alleviate pathological injury and regulate the coagulation function to promote recovery of the endometrium. Angelica water extract could increase the levels of E2, Pg, and HCG and the expression of ERα, Erβ, and PR. Angelica water extract could regulate the metabolic pathways which include the glycerophospholipid metabolism, sphingolipid metabolism, glycine, serine and threonine metabolism, and retinol metabolism.

Parkinson's disease (PD) is one of the most common degenerative diseases of the human nervous system and has a wide range of serious impacts on human health and quality of life. Recently, research targeting high mobility group box 1 (HMGB1) in PD has emerged, and a variety of laboratory methods for inhibiting HMGB1 have achieved good results to a certain extent. However, given that HMGB1 undergoes a variety of intracellular modifications and three different forms of extracellular redox, the possible roles of these forms in PD are likely to be different. General inhibition of all forms of HMGB1 is obviously not ideal and has become one of the biggest obstacles in the clinical application of targeting HMGB1. In this review, pure mechanistic research of HMGB1 and in vivo research targeting HMGB1 were combined, the effects of HMGB1 on neurons and immune cell responses in PD are discussed in detail, and the problems that need to be focused on in the future are addressed.

Deslagging of Eichhornia crassipers and Pistia stratiotes biomass pellets was studied after dynamic or static addition of CaO, MgO or kaoline at 600 or 800 °C. Then the chemical element and crystal phase compositions of ashes were characterized by X-ray fluorescence and X-ray diffraction. The K and Cl concentrations declined by 10%–12% and 6%–8% respectively after the use of additives, and were both higher under dynamic addition than under static addition at the same temperature. The ashes contained some high-melting-point crystal phases (e.g. Ca3Si2O7, CaMgSi2O7), which can raise the melting temperature of ashes. By comparing the slagging indices between the two types of biomass ashes, we found the deslagging effect of static loading of additives was better than dynamic loading. The deslagging effect was optimized after the static loading of kaoline at 800 °C for both types of biomass. Under the condition that these research results are applied to thermal system of power plant, it can provide basis for solving slagging problem of biomass boiler.

Cyclin-dependent kinases (CDKs) are hyperactive in many cancers and have served as cancer therapeutic targets for decades. Palbociclib (Palb) is the first approved CDK4/6 inhibitor to treat hormone receptor (HR)-positive, HER2-negative advanced breast cancer. Acquired drug resistance is one obstacle of Palb be utilized in other cancer. CDK2 compensation of CDK4/6 loss is one of the causes that cancer cells are resistant to Palb. Hence, targeting multiple CDKs could be a novel strategy to prevent the drug resistance of cancer cells and expand the application of Palb in other cancer. In this study, we initially indicated Polyphyllin I (PPI) significantly inhibits non-small lung cancer cell (NSCLC) proliferation, promotes cell apoptosis in vitro and in vivo. Mechanistically, PPI can inhibit Rb through the p21/CDK2/Rb signaling pathway in NSCLC. A combination of PPI and Palb exerts a significant synergistic anti-cancer ability on NSCLC. Of note, PPI can reverse Palb drug resistance. Herein, we first time demonstrated PPI can disturb CDK2 function through upregulation of p21. The PPI effect on CDK2 provides a choice for a chemotherapeutic strategy for the elimination of NSCLC. Our study highlighted the clinical significance of simultaneously blocking of CDK2 and CDK4/6 for NSCLC treatment.

Microbial enzymes are crucial for material biotransformation during the composting process. In this study, we investigated the effects of adding bamboo charcoal (BC) (i.e., at 5%, 10%, and 20% corresponding to BC5, BC10, and BC20, respectively) on the enzyme activity levels during chicken manure composting. The results showed that BC10 could increase the cellulose and urease activities by 56% and 96%, respectively. The bacterial community structure in BC10 differed from those in the other treatments, and Luteivirga, Lactobacillus, Paenalcaligenes, Ulvibacter, Bacillus, Facklamia, Pelagibacterium, Sporosarcina, Cellvibrio, and Corynebacterium had the most important roles in composting. Compared with other treatments, BC10 significantly enhanced the average rates of degradation of carbohydrates (D-xylose (40%) and α-D-lactose (44%)) and amino acids (L-arginine (16%), L-asparagine (14%), and L-threonine (52%)). We also explored the associations among the bacterial community and their metabolic functions with the changes in the activities of enzymes. Network analysis demonstrated that BC10 altered the co-occurrence patterns of the bacterial communities, where Ulvibacter and class Bacilli were the keystone bacterial taxa with high capacities for degrading carbon source, and they were related to increases in the activities of cellulase and urease, respectively. The results obtained in this study may help to further enhance the efficiency of composting.

Epidemiological studies suggest that men exhibit a higher mortality rate to COVID-19 than women, yet the underlying biology is largely unknown. Here, we seek to delineate sex differences in the gene expression of viral entry proteins ACE2 and TMPRSS2, and host transcriptional responses to SARS-CoV-2 through large-scale analysis of genomic and clinical data. We first compiled 220,000 human gene expression profiles from three databases and completed the meta-information through machine learning and manual annotation. Large scale analysis of these profiles indicated that male samples show higher expression levels of ACE2 and TMPRSS2 than female samples, especially in the older group (>60 years) and in the kidney. Subsequent analysis of 6,031 COVID-19 patients at Mount Sinai Health System revealed that men have significantly higher creatinine levels, an indicator of impaired kidney function. Further analysis of 782 COVID-19 patient gene expression profiles taken from upper airway and blood suggested men and women present distinct expression changes. Computational deconvolution analysis of these profiles revealed male COVID-19 patients have enriched kidney-specific mesangial cells in blood compared to healthy patients. Together, this study suggests biological differences in the kidney between sexes may contribute to sex disparity in COVID-19.

Fagopyrum dibotrys (F. dibotrys) (D.Don) H.Hara is a well-known edible herbal medicine in Asian countries. It has been widely used for the treatment of lung diseases, swelling, etc., and is also an important part of many Chinese medicine prescriptions. At present, more than 100 compounds have been isolated and identified from F. dibotrys, and these compounds can be primarily divided into flavonoids, phenols, terpenes, steroids, and fatty acids. Flavonoids and phenolic compounds are considered to be the main active ingredients of F. dibotrys. Previous pharmacological studies have shown that F. dibotrys possesses anti-inflammatory, anti-cancer, anti-oxidant, anti-bacterial, and anti-diabetic activities. Additional studies on functional genes have led to a better understanding of the metabolic pathways and regulatory factors related with the flavonoid active ingredients in F. dibotrys. In this paper, we systemically reviewed the research advances on the phytochemistry and pharmacology of F. dibotrys, as well as the functional genes related to the synthesis of active ingredients, aiming to promote the development and utilization of F. dibotrys.

Streptococcus suis is an important zoonotic pathogen. An increase in multi-drug-resistant strains has led to poor performance of traditional antibiotic therapies. Thus, alternative antibacterial agents are urgently needed. In this study, we identified a recombined and expressed lysin PlyARI derived from the novel serotype S. suis (Chz) prophage PhiARI0460-1. The recombinant PlyARI at a concentration of 10 µg/mL showed high bacteriolytic activity against 30 S. suis isolates. The minimum inhibitory concentration (MIC) of PlyARI against S. suis was found to be as low as 2 µg/mL, and the lytic efficiency could be maintained between the range of pH 4 and 12. Additionally, in a mouse infection model, a dose of 0.5 mg of PlyARI protected 10 out of 10 mice that were challenged with highly virulent S. suis strain HA9801. Furthermore, the binding specificity of PlyARI was evaluated by constructing a green fluorescent protein (GFP-ARIb), where GFP was fused with the PlyARI-SH3b (cell wall-binding domain, CBD), revealing a high affinity to S. suis, Staphylococcus aureus, and Streptococcus equi along with exhibiting a medium affinity to Streptococcus pneumoniae as well as Streptococcus agalactiae. Overall, our findings indicated that PlyARI may be an alternative antibacterial agent that was useful in treating and possibly the prevention of Streptococcal infections.

Photocatalysis is regarded as one of the most promising technologies to remove organic contaminants. At present, most of the covalent organic frameworks (COFs) used as photocatalysts are connected by imine or borate bonds, which have relatively low stability and relatively poor π-delocalization. Herein, we report, for the first time, vinylene-linked COFs constructed by various diacetylene and triazine moieties for photocatalytic degradation of organic contaminants and disinfection of bacteria. The pioneering introduction of diacetylene moieties not only enhances conjugated π-electrons delocalization but also optimizes the electronic band structures that significantly improve photocatalytic activity. Therefore, the vinylene-bridged COFs have excellent photocatalytic activity with ultrahigh stability and great π-electron delocalization, thus exhibiting ultrafast photocatalytic degradation efficiency for phenol and norfloxacin (96%, within 15 min). Our work provides a strong basis for the rational regulation of the chemical structure of COFs to enhance their photocatalytic activity, thus broadening the application of COFs in photocatalysis.

Polymyxin B and colistin (polymyxin E) are increasingly used as the last line of therapy for infections caused by multidrug-resistant (MDR) gram-negative pathogens. However, nephrotoxicity is still...

Jie Wang,1,2 Yun Wu,3 Md Nazim Uddin,2,4 Rong Chen,5 Jian-Ping Hao5 1Department of Pharmacy, First Affiliated Hospital of Xinjiang Medical University, Urumqi, 830011, People’s Republic of China; 2School of Basic Medicine and Clinical Pharmacy, China Pharmaceutical University, Nanjing, 211198, People’s Republic of China; 3Department of General Medicine, First Affiliated Hospital of Xinjiang Medical University, Urumqi, 830011, People’s Republic of China; 4Bangladesh Council of Scientific and Industrial Research (BCSIR), Dhaka, 1205, Bangladesh; 5Department of Hematology, First Affiliated Hospital of Xinjiang Medical University, Urumqi, 830011, People’s Republic of ChinaCorrespondence: Jian-Ping Hao No. 137 Liyushan S. Road, Urumqi, Xinjiang, People’s Republic of ChinaTel +86 13579876416Email 13579876416@163.comIntroduction: Essential thrombocytosis (ET) is a group of myeloproliferative neoplasms characterized by abnormal proliferation of platelet and megakaryocytes. Research on potential key genes and novel regulatory markers in essential thrombocythemia (ET) is still limited.Methods: Downloading array profiles from the Gene Expression Omnibus database, we identified the differentially expressed genes (DEGs) through comprehensive bioinformatic analysis. GO, and REACTOME pathway enrichment analysis was used to predict the potential functions of DEGs. Besides, constructing a protein–protein interaction (PPI) network through the STRING database, we validated the expression level of hub genes in an independent cohort of ET, and the transcription factors (TFs) were detected in the regulatory networks of TFs and DEGs. And the candidate drugs that are targeting hub genes were identified using the DGIdb database.Results: We identified 63 overlap DEGs that included 21 common up-regulated and 42 common down-regulated genes from two datasets. Functional enrichment analysis shows that the DEGs are mainly enriched in the immune system and inflammatory processes. Through PPI network analysis, ACTB, PTPRC, ACTR2, FYB, STAT1, ETS1, IL7R, IKZF1, FGL2, and CTSS were selected as hub genes. Interestingly, we found that the dysregulated hub genes are also aberrantly expressed in a bone marrow cohort of ET. Moreover, we found that the expression of CTSS, FGL2, IKZF1, STAT1, FYB, ACTR2, PTPRC, and ACTB genes were significantly under-expressed in ET (P< 0.05), which is consistent with our bioinformatics analysis. The ROC curve analysis also shows that these hub genes have good diagnostic value. Besides, we identified 4 TFs (SPI1, IRF4, SRF, and AR) as master transcriptional regulators that were associated with regulating the DEGs in ET. Cyclophosphamide, prednisone, fluorouracil, ruxolitinib, and lenalidomide were predicted as potential candidate drugs for the treatment of ET.Discussion: These dysregulated genes and predicted key regulators had a significant relationship with the occurrence of ET with affecting the immune system and inflammation of the processes. Some of the immunomodulatory drugs have potential value by targeting ACTB, PTPRC, IL7R, and IKZF1 genes in the treatment of ET.Keywords: essential thrombocythemia, hub genes, regulatory markers, candidate drugs, bioinformatics analysis

Severe COVID-19 disease caused by SARS-CoV-2 is frequently accompanied by dysfunction of the lungs and extrapulmonary organs. However, the organotropism of SARS-CoV-2 and the port of virus entry for systemic dissemination remain largely unknown. We profiled 26 COVID-19 autopsy cases from four cohorts in Wuhan, China, and determined the systemic distribution of SARS-CoV-2. SARS-CoV-2 was detected in the lungs and multiple extrapulmonary organs of critically ill COVID-19 patients up to 67 days after symptom onset. Based on organotropism and pathological features of the patients, COVID-19 was divided into viral intrapulmonary and systemic subtypes. In patients with systemic viral distribution, SARS-CoV-2 was detected in monocytes, macrophages, and vascular endothelia at blood–air barrier, blood–testis barrier, and filtration barrier. Critically ill patients with long disease duration showed decreased pulmonary cell proliferation, reduced viral RNA, and marked fibrosis in the lungs. Permanent SARS-CoV-2 presence and tissue injuries in the lungs and extrapulmonary organs suggest direct viral invasion as a mechanism of pathogenicity in critically ill patients. SARS-CoV-2 may hijack monocytes, macrophages, and vascular endothelia at physiological barriers as the ports of entry for systemic dissemination. Our study thus delineates systemic pathological features of SARS-CoV-2 infection, which sheds light on the development of novel COVID-19 treatment.

Rapid methanogenesis recovery of an acidified anaerobic digestion (AD) system is a challenging issue in engineering application, due to the unfavorably thermodynamics of volatile fatty acids (VFA) syntrophic oxidation process with hydrogen as electron transfer mediator. To breakthrough this bottleneck, in this study, we developed the strategy with biochar as an additive for rapid methanogenesis recovery of a severely acidified AD system is elucidated. First, with VFA as substrate, it was found that biochar addition shortened lag time and promoted maximum methane production rate, confirming the role of biochar to enhance VFA syntrophic oxidation. Moreover, the addition of biochar to the acidified sludge significantly decreased hydrogen partial pressure, which thermodynamically stimulated VFA oxidation by reducing Gibbs free energy. Microbial community analysis delivered that biochar addition largely altered the dominant microbes. The enrichment of electro-active Syntrophomonas and Methanosarcina suggested the role of biochar as potential redox-active mediator to stimulate potential direct interspecies electron transfer between syntrophic microorganisms, and inhibit hydrogen-based syntrophic pathway simultaneously. Further stable operation confirms that biochar is a promising additive for the rapid methanogenic recovery of acidified AD systems.

Skeletal muscle differentiation is a highly coordinated process that involves many cellular signaling pathways and microRNAs (miRNAs). A group of muscle-specific miRNAs has been reported to promote myogenesis by suppressing key signaling pathways for cell growth. However, the functional role and regulatory mechanism of most non-muscle-specific miRNAs with stage-specific changes during differentiation are largely unclear. Here, we describe the functional characterization of miR-101a/b, a pair of non-muscle-specific miRNAs that show the largest change among a group of transiently upregulated miRNAs during myogenesis in C2C12 cells. The overexpression of miR-101a/b inhibits myoblast differentiation by suppressing the p38/MAPK, Interferon Gamma, and Wnt pathways and enhancing the C/EBP pathway. Mef2a, a key protein in the p38/MAPK pathway, was identified as a direct target of miR-101a/b. Interestingly, we found that the long non-coding RNA (lncRNA) Malat1, which promotes muscle differentiation, interacts with miR-101a/b, and this interaction competes with Mef2a mRNA to relieve the inhibition of the p38/MAPK pathway during myogenesis. These results uncovered a “braking” role in differentiation of transiently upregulated miRNAs and provided new insights into the competing endogenous RNA (ceRNA) regulatory mechanism in myoblast differentiation and myogenesis.

The aim of the present study is to explored the relationship between ADIPO signalling pathway and T2DM, to provide clues for further study of the pathogenesis of T2DM and to determine the possible drug targets. This study employed a case-control study design. Twenty-three single nucleotide polymorphisms (SNPs) of 13 genes in the selected ADIPO signalling pathway were genotyped by SNPscanTM kit. All statistical analysis was performed by SPSS 25.0, PLINK 1.07, R 2.14.2, Haploview 4.2, SNPstats, and other statistical software packages. In the association analysis based on a single SNPs, rs1044471 had statistical significance in the overdominant model without adjusting covariates. Rs1042531 had statistical significance in the overdominant model. Rs12718444 had statistical significance in the recessive model. There was a linkage disequilibrium between the loci within 9 genes, and the two loci in RXRA gene did not form blocks. Four kernel functions were used for SNPs set analysis based on ADIPO signalling pathway showed that there was no statistical significance whether covariates were added or not, P>0.05.According to our research results, it is found that some single nucleotide polymorphisms (ADIPOR2 rs1044471, PCK1 rs1042531, GLUT1 rs12718444) in the adiponectin signalling pathway may be associated with T2DM

Brassica rapa is one of the most important leafy vegetable crops with large cultivated area in China. To increase the availability of DNA markers in B. rapa, we developed insertion-deletion (InDel) markers utilizing high-resolution melting (HRM) curve analysis. We designed primers for 252 InDels (≥ 3 bp) evenly distributed in the genome and tested gene polymorphisms with eight accessions. In total, 208 markers were specifically amplified, and 148 InDels with polymorphism were genotyped successfully using HRM. We further analyzed the correlation with InDel size, GC number, and predicted the difference in Tm values (∆Tm) using 208 markers with specific amplification. We found that the success rate of InDel markers was correlated with the GC number of InDel and the predicted-∆Tm, but not clearly correlated with the length of InDel. When the GC number within InDel was ≥ 8, the successful rate exceeded 90.0%. When the predicted-∆Tm reached 0.5 °C, the success rate was greater than 90.0%, and when it was ≥ 0.6 °C, the rate climbed to 100.0%, indicating their role as the optimal parameter for successful development of an applicable InDel marker. The polymorphic InDel markers can be easily genotyped using HRM. They are of great value in genetic analysis, construction of linkage map, and molecular marker-assisted selection in B. rapa.

Background Gastric cancer is one of the Lynch syndrome (LS)-associated malignancies. Previous studies have suggested that LS patients with gastric cancer also had chronic atrophic gastritis in the background mucosa, but further histologic characterization was not attempted. This study aims to understand the histologic features of background chronic gastritis in LS patients with gastric adenocarcinoma. Methods Eleven LS-associated gastric cancer cases were collected from five institutions. Demographics and clinical features were retrieved by review of medical charts. Pathological material was reviewed for tumor location and histologic type. In addition, non-neoplastic gastric mucosa was assessed for inflammation (chronic and active), atrophy, intestinal metaplasia (IM) in the antrum and body, as well as pyloric gland metaplasia and enterochromaffin-like (ECL) cell hyperplasia in the body. Results Eleven LS patients with gastric cancer (four male and seven female) with a mean age of 63 years (range: 23 - 83) were included. Ten (90.9%) had personal cancer histories; however none of the patients had family history of gastric cancer. Eight (72.7%) patients underwent gastrectomy and three had endoscopic resection. Nine (81.8%) patients had tumor in the fundus and/or body and two had tumor present in the antrum. Seven (63.6%) cases were intestinal type or mixed type carcinoma, and the remaining four were signet ring cell carcinoma. Eight (of 11, 72.7%) patients had chronic gastritis, five (45.4%) had atrophy, and four (36.3%) had intestinal metaplasia. Four of five patients with both antrum and body mucosa available for evaluation (80%), demonstrated body-predominant chronic gastritis. Four patients had germline MLH1 alterations and all of these patients had chronic gastritis, including one Helicobacter pylori (H. pylori) gastritis and three H. pylori-negative gastritis. Conclusions None of LS patients with gastric cancer in our cohort had a family history of gastric cancer. Gastric adenocarcinomas in LS patients were primarily located in the fundus and/or body. Two-thirds of these tumors were of intestinal type and had a background chronic, H. pylori-negative gastritis. These results support a chronic atrophic gastritis with intestinal metaplasia-dysplasia-carcinoma sequence in LS-related gastric tumorigenesis, particularly in MLH1-mutated LS patients.

Single-stage partial nitration and anammox (PN/A) is achieved using a submerged dynamic membrane (DM-SBR) in this study. The DM-SBR was stably operated for 200 days, and the nitrogen removal efficiency (NRE) was sustained at 70.3 ± 7.2% at a nitrogen loading rate (NLR) ranging from 0.1 to 0.3kgN/m3/d with a hydraulic retention time (HRT) of 24 h. When the NLR was 0.2 kgN/m3/d, the NRE achieved was high as 80% with a low concentration of dissolved oxygen (DO) of 0.13 mg/L. In addition, the specific activity of anammox bacteria (AnAOB) and ammonia-oxidizing bacteria (AOB) reached was 2.72 and 16.80 gN/gVSS/d, respectively. The dynamic membrane (DM) intercepted the biomass due to the lamellar, intact, dense biofilm self-generated on the surface of the supporting material, which had an effluent turbidity of 10 NTU. The enriched anammox functional bacteria were Candidatus Jettenia, and the relative abundance was 11.06%. The AOB-like functional bacteria consisted primarily of Nitrosomonas, with a relative abundance of 2.76%, which ensured the nitrogen removal process reaction. This study provides a novel reactor configuration of the single-stage PN/A process in the view of practical applications.

Purpose: Whole-exome (WES) and RNA sequencing (RNA-seq) are key components of cancer immunogenomic analyses. To evaluate the consistency of tumor WES and RNA-seq profiling platforms across different centers, the Cancer Immune Monitoring and Analysis Centers (CIMAC) and the Cancer Immunologic Data Commons (CIDC) conducted a systematic harmonization study. Experimental Design: DNA and RNA were centrally extracted from fresh frozen and formalin-fixed paraffin-embedded non–small cell lung carcinoma tumors and distributed to three centers for WES and RNA-seq profiling. In addition, two 10-plex HapMap cell line pools with known mutations were used to evaluate the accuracy of the WES platforms. Results: The WES platforms achieved high precision (> 0.98) and recall (> 0.87) on the HapMap pools when evaluated on loci using > 50× common coverage. Nonsynonymous mutations clustered by tumor sample, achieving an index of specific agreement above 0.67 among replicates, centers, and sample processing. A DV200 > 24% for RNA, as a putative presequencing RNA quality control (QC) metric, was found to be a reliable threshold for generating consistent expression readouts in RNA-seq and NanoString data. MedTIN > 30 was likewise assessed as a reliable RNA-seq QC metric, above which samples from the same tumor across replicates, centers, and sample processing runs could be robustly clustered and HLA typing, immune infiltration, and immune repertoire inference could be performed. Conclusions: The CIMAC collaborating laboratory platforms effectively generated consistent WES and RNA-seq data and enable robust cross-trial comparisons and meta-analyses of highly complex immuno-oncology biomarker data across the NCI CIMAC-CIDC Network.

Wen Cao,1 Cong Zhang,1 Rong Chen,2,3 Qianqian Wu,1 Renhao Xu,2,3 Lan Zhang,1 Xiangjian Zhang1– 3 1Department of Neurology, Second Hospital of Hebei Medical University, Shijiazhuang, Hebei 050000, People’s Republic of China; 2Hebei Collaborative Innovation Center for Cardio-Cerebrovascular Disease, Shijiazhuang, Hebei 050000, People’s Republic of China; 3Hebei Vascular Homeostasis Key Laboratory for Neurology, Shijiazhuang, Hebei 050000, People’s Republic of ChinaCorrespondence: Xiangjian ZhangDepartment of Neurology, Second Hospital of Hebei Medical University, 215 Hepingxi Road, Shijiazhuang, Hebei 050000, People’s Republic of ChinaTel +86 15803210578Fax +86 31166002822Email zhang666xj@sina.comPurpose: Strokes are devastating as there are no current therapies to prevent long-term neurological deficits. Previous studies reported that cerebroprotein hydrolysate (CH) plays a role in neuronal protection in acute phase after ischemic stroke, while the long-term effects of CH upon brain plasticity and neurological outcomes after stroke are still uncertain. To address these gaps, we assessed the effect of a new cerebroprotein hydrolysate, CH1, on long-term gray and white matter integrity as well as axonal plasticity in the late phase after ischemic stroke and the potential mechanisms.Methods: Adult male mice were subjected to permanent distal middle cerebral artery occlusion (dMCAO), followed by daily intraperitoneal injection of CH1 for 14 days. Motor function was measured weekly through behavioral neurological evaluations. Gray matter intensity and white matter intensity were examined by immunofluorescence staining. The sonic hedgehog (Shh) inhibitor cyclopamine (CYC) was injected to determine the involvement of the Shh pathway in the therapeutic effects of CH1.Results: We found that intraperitoneal delivery of CH1, compared to vehicle administration, significantly improved long-term neurological outcomes at various times and promoted neuronal viability at 14 days but not at 28 days after stroke. Importantly, CH1 mitigated stroke-induced white matter injury and facilitated axonal plasticity in the late stage after stroke.Conclusion: These results unveil a previously unappreciated role for CH in the repair of white matter and brain plasticity after stroke.Keywords: ischemic stroke, cerebroprotein hydrolysate, axonal plasticity, white matter integrity, Shh pathway

Two new dimeric abietane-type diterpenoids, 12-hydroxyabieta-8,11,13-trien-7-α-yl 7-oxoabieta-8,11,13-trien-12-yl peroxide (trivial name japonicinol C, 1) and 12-hydroxyabieta-8,11,13-trien-7-α-yl 7-oxoabieta-5,8,11,13-tetraen-12-yl peroxide (trivial name japonicinol D, 3), together with two known dimeric abietane-type diterpenoids, obtusanol A (2) and 7-α-(2-butoxyethoxy)-12-hydroxyabieta-6-yl 6,7-dehydroabieta-8,11,13-trien-12-yl ether (4), were isolated from the methanol extract of the bark of Cryptomeria japonica. Their structures were established by mean of spectroscopic analysis and comparison of NMR data with those of known analogues. At a concentration of 50 μM, compounds 1–4 inhibited xanthine oxidase activity by 29.8, 39.3, 27.1, and 14.7 %, respectively. In addition, compounds 1 and 4 also showed 13.9 and 20.5 % inhibition toward angiotensin-converting enzyme (ACE), respectively.

In this paper, water hyacinth and Pistia stratiotes pellet fuel are the research objects, and the flue gas emission characteristics of aquatic biomass compact fuels burning in the O2/CO2 atmosphere...

Wei-Hua Xiao,1 Li-Ping Yao,2 Min Li,2 Min Wang,1 Liang Wu,2 Mao-Fen Jiang,1 Hai-Fen Ma,1 Jun-Qiang Li,1 Guo-Rong Chen2 1Department of Pathology, Ningbo Beilun People’s Hospital, Ningbo 315800, People’s Republic of China; 2Department of Pathology, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, People’s Republic of ChinaCorrespondence: Wei-Hua XiaoDepartment of Pathology, Ningbo Beilun People’s Hospital, Ningbo 315800, People’s Republic of ChinaTel +86 0574 8610 0266Fax +86 0574 8677 6335Email xiaoweihua_dr66@163.comGuo-Rong ChenDepartment of Pathology, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, People’s Republic of ChinaTel/Fax +86 0577 8668 9908Email chengr_dr3366@163.comObjective: The aim of this study was to investigate the effect of the tumor-associated macrophage-m2-cancer cell complex (TAM-M2-CC) on the heterostructural modification of lung adenocarcinoma.Methods: The expression of CD163+/CD68+ in macrophages in the microenvironment of 161 cases of lung adenocarcinoma was identified by dual immunohistochemistry, and the association between a TAM-M2-CC and its growth, as well as the histological changes in lung adenocarcinoma cells, was assessed.Results: The morphological change of lung adenocarcinoma was related to the number of m2 phenotypes of the macrophages in the microenvironment of lung adenocarcinoma. TAM-M2-CCs were involved in the process of cancer cell recognition, association, and reconstruction.Conclusion: The microenvironment of lung adenocarcinoma can affect the phenotypic distinction of macrophages, and the polarization recruitment, zombification, and formation of a TAM-M2-CC, which can also affect the local differentiation of lung adenocarcinoma to a certain extent. The applicable pathogenesis needs to be verified and studied further.Keywords: tumor-related macrophage-M2-cancer cell complex, TAM-M2-CC, heterostructural modification, microenvironment of lung adenocarcinoma, phenotypic differentiation, local differentiation

Phenylalanine hydroxylase (PAH) deficiency leads to phenylalanine accumulation and results in phenylketonuria (PKU). Phenylketonuria can contribute to severe inability such as mental impairment. Early diagnosis and dietary intervention can have beneficial effects on maintaining normal neural and cognitive function in patients with PKU. However, a long-term low phenylalanine diet may put children at risk of malnutrition. A food supplement was therefore used for children with PKU under dietician supervision according to dietary reference intakes (DRIs). In this cross-sectional study, we enrolled patients with PKU and age-matched controls to compare their anthropometry data [weight, height, body mass index (BMI), and body composition using bioelectrical impedance analysis (BIA)], and correlated it with their dietary intake based on 24-h dietary recall. For continuous parameters, the data were expressed as median ± standard deviation (SD), and the Mann–Whitney U test was used to test the difference among the groups. Correlation by natural proteins, body fat, and fat-free mass were evaluated using the Pearson correlation coefficient. Twenty-two participants diagnosed with PKU (ages 8–27 years; mean 15.23 ± 5.23) and a control group of 22 non-PKU participants (ages 8–39 years; mean 19.73 ± 10.6) were recruited for this study. Between the two groups of participants, no significant difference was found in height, weight, BMI, muscle mass, or fat mass. The percentage of natural protein has no effect on body composition. We found a significant positive correlation between the total protein intake percentage of DRIs and muscle mass (r = 0.491, p = 0.020) and a significant negative correlation in the total protein intake percentage of DRIs and fat mass (r = -0.475, p = 0.025) in participants with PKU. There were no significant differences in body composition and nutrition intake between patients with PKU (under metabolic control) and healthy subjects. Thus, giving proper nutrition treatment may have beneficial effects on body growth and nutrition status in patients with PKU in Taiwan.

Polysaccharide was derived from Pueraria lobata (PPL) which was considered as one of the traditional Chinese medicinal and edible herbs. In the present study, PPL was administered in equal doses (12.5 mg/kg) to both normal mice and antibiotic-associated diarrhea (AAD) mice for two weeks, and was evaluated in terms of body weight, organ indices, gut structure, gut microbiota and short chain fatty acids. The results showed that normal mice treated with PPL not only reduced the isovaleric acid concentration (P 0.05), but also significantly increased the abundance of beneficial bacteria, involving Oscillospira and Anaerotruncus (P 0.05). In addition, PPL could relieve colonic pathological changes and gut microbiota dysbiosis caused by AAD. It indicated that PPL was a potential functional food ingredient by modulating gut microbiota.

Nelumbo nucifera leaves extracts (NLE) has been suggested to provide antioxidant, anti-obesity and anticancer effects. However, the research on the anti-melanogenetic effect of NLE was little. Here, we reported that NLE and Gallic acid (GA, major ingredients of NLE) exhibit the hypopigmentary effect in B16F1 cells. NLE and GA showed potent inhibitory effects on tyrosinase, microphthalmia-associated transcription factor (MITF), and tyrosinase-related protein-1 (TRP-1) protein production. The phosphorylation of intracellular protein kinase A (PKA) and cAMP response element-binding protein (CREB) were also decreased, revealing potent anti-melanogenic effects of NLE and GA. However, NLE showed the better effect than GA on reducing melanin formation, implying the importance of the synergism of polyphenolic compounds of NLE. Furthermore, NLE inhibited skin melanogenesis and epidermal hyperplasia of guinea pigs caused by irradiation through attenuating ERK and CREB activation. NLE reduced skin melanogenesis and epidermal hyperplasia induced by UVB in guinea pigs through downregulation ERK and CREB pathways, and the following decreasing of MITF, tyrosinase and TRP-1 expression. These results demonstrated the potential which NLE as an ingredient of hypopigmentary cosmetics.

A cucurbitane triterpenoid, 7β-butoxy-3β-hydroxy-25-methoxycucurbita-5,23(E)-dien-19-al (1), was isolated from the fruit pulp of Momordica charantia. Structure determination of the new compound was accomplished by spectroscopic analyses including 1D and 2D NMR (1H, 13C, COSY, HMQC, HMBC, and NOESY) and EI-MS and comparison with the data of known analogues.

Herein we develop a unique differentiated-uptake strategy capable of efficient and high-purity isolation of genuine drug-resistant (DR) cells from three types of drug-surviving cancer cells, which include paclitaxel-surviving human ovarian OVCAR-3 cancer cells and human lung carcinoma A549/Taxol cells, and doxorubicin-surviving human immortalized myelogenous leukemia K562/ADR cells. By using this strategy which relies on fluorescent glycan nanoparticle (FGNP)-based fluorescence-activated cell sorting (FACS) assays, two subpopulations with distinct fluorescences existing in drug-surviving OVCAR-3 cells were separated, and we found that the lower fluorescence (LF) subpopulation consisted of DR cells, while the higher fluorescence (HF) subpopulation was comprised of non-DR cells. Besides, the DR cells and their progenies were found distinct in their increased expression of drug-resistant genes. More intriguingly, by using the FGNP-based FACS assay to detect DR/non-DR phenotypes, we found that the DR phenotype had a potential to differentiate into the non-DR progeny, which demonstrates the differentiation feature of stem-like cancer cells. Further research disclosed that the assay can quantitatively detect the degree of drug resistance in DR cells, as well as the reversal of drug resistance that are tackled by various therapeutic methods. The strategy thus paves the way to develop theranostic approaches associated with chemotherapy-resistance and cancer stemness.

Ralstonia solanacearum species complex is the most destructive and economically important bacterial pathogen of many plant species around the world. It is a particular constraint to the production of the solanaceous vegetables such as tomato, chilli pepper and African eggplant in West Africa. Its broad host range, ability to survive in the soil for long periods and ability to sustain latent infection, make it difficult to control. Identification, characterization and mapping the distribution of the causal Ralstonia spp. strains are necessary to help design effective control strategies. Wilted tomato, African eggplant and pepper plant samples were collected from fields in different regions of Mali. The causal bacterial strains were isolated from the samples and all were identified as Ralstonia pseudosolanacearum. Multiplex PCR using four phylotype specific primer pairs identified the presence of phylotype I strains distributed across all the vegetable production areas whereas phylotype III strains were limited to areas of Sikasso, Koulikoro and Segou to the south and east of Bamako city. Phylogenetic analysis of part of the conserved endoglucanase virulence gene sequences revealed four sequevars with most phylotype I strains identified as sequevars 46 and 31 with only a few as sequevar 14 and one sequevar 18. The phylotype III strains were all in the sequevar 23–48 group. Pathogenicity testing of a selected subset of the strains isolated in Mali showed them all to be pathogenic to the susceptible tomato cultivar Roma. To our knowledge, this is the first study on the molecular diversity within the Ralstonia spp. complex in Mali. This information is important for developing wilt management strategies including breeding for resistance and specifying quarantine restrictions.

Joubert syndrome is characterized by unique malformation of the cerebellar vermis. More than thirty Joubert syndrome genes have been identified, including ARL13B. However, its role in cerebellar development remains unexplored. We found that knockdown or knockout of arl13b impaired balance and locomotion in zebrafish larvae. Granule cells were selectively reduced in the corpus cerebelli, a structure homologous to the mammalian vermis. Purkinje cell progenitors were also selectively disturbed dorsomedially. The expression of atoh1 and ptf1, proneural genes of granule and Purkinje cells, respectively, were selectively down-regulated along the dorsal midline of the cerebellum. Moreover, wnt1, which is transiently expressed early in cerebellar development, was selectively reduced. Intriguingly, activating Wnt signaling partially rescued the granule cell defects in arl13b mutants. These findings suggested that Arl13b is necessary for the early development of cerebellar granule and Purkinje cells. The arl13b-deficient zebrafish can serve as a model organism for studying Joubert syndrome.

Astrocytoma is the most common type of primary brain tumor. The risk factors for astrocytoma are poorly understood; however, germline genetic variants account for 25% of the risk of developing gliomas. In this study, we assessed the risk of astrocytoma associated with variants in AGT, known by its role in angiogenesis, TP53, a well-known tumor suppressor and the DNA repair gene MGMT in a Mexican population. A case-control study was performed in 49 adult Mexican patients with grade II-IV astrocytoma. Sequencing of exons and untranslated regions of AGT, MGMT, and TP53 from was carried in an Ion Torrent platform. Individuals with Mexican Ancestry from the 1000 Genomes Project were used as controls. Variants found in our cohort were then assessed in a The Cancer Genome Atlas astrocytoma pan-ethnic validation cohort. Variants rs1926723 located in AGT (OR 2.74, 1.40-5.36 95% CI), rs7896488 in MGMT (OR 3.43, 1.17-10.10 95% CI), and rs4968187 in TP53 (OR 2.48, 1.26-4.88 95% CI) were significantly associated with the risk of astrocytoma after multiple-testing correction. This is the first study where the AGT rs1926723 variant, TP53 rs4968187, and MGMT rs7896488 were found to be associated with the risk of developing an astrocytoma.

The 1,3-enyne moiety is commonly found in cyclohexanoid natural products produced by endophytic and plant pathogenic fungi. Asperpentyn (1) is a 1,3-enyne-containing cyclohexanoid terpenoid isolated from Aspergillus and Pestalotiopsis. The genetic basis and biochemical mechanism of 1,3-enyne biosynthesis in 1, and other natural products containing this motif, has remained enigmatic despite their potential ecological roles. Identified here is the biosynthetic gene cluster and characterization of two crucial enzymes in the biosynthesis of 1. A P450 monooxygenase that has a dual function, to first catalyze dehydrogenation of the prenyl chain to generate a cis-diene intermediate and then serve as an acetylenase to yield an alkyne moiety, and thus the 1,3-enyne, was discovered. A UbiA prenyltransferase was also characterized and it is unusual in that it favors transferring a five-carbon prenyl chain, rather than a polyprenyl chain, to a p-hydroxybenzoic acid acceptor.

Colorectal cancer (CRC) is a common malignancy. Many reports have implicated aberrant mitochondrial activity in the progression of CRC, with particular emphasis on the dysregulation of redox signaling and oxidative stress. In this study, we focused on manganese superoxide dismutase (MnSOD/SOD2), a key antioxidant enzyme, which maintains intracellular redox homeostasis. Current literature presents conflicting mechanisms for how SOD2 influences tumorigenesis and tumor progression. Here, we explored the role of SOD2 in CRC specifically. We found high levels of SOD2 expression in CRC tissues. We carried out a series of experiments to determine whether knockdown of SOD2 expression in CRC cell lines would reverse features of tumorigenesis. We found that reduced SOD2 expression decreased cell proliferation, migration, and invasion activity in CRC cells. Results from an additional series of experiments on mitochondrial function implicated a dual role for SOD2 in promoting CRC progression. First, proper level of SOD2 helped CRC cells maintain mitochondrial function by disposal of superoxide (O2 .- ). Second, over-expression of SOD2 induced H2 O2 -mediated tumorigenesis by upregulating AMPK and glycolysis. Our results indicate that SOD2 may promote the occurrence and development of CRC by regulating the energy metabolism mediated by AMPK signaling pathways.

Bioreductions catalyzed by alcohol dehydrogenases (ADHs) play an important role in the synthesis of chiral alcohols. However, the synthesis of ethyl (S)-4-chloro-3-hydroxybutyrate [(S)-CHBE], an im...

Thermo-sensitive genic male-sterile (TGMS) gene tms5 is most widely used in the two-line hybrid breeding system in China. To develop novel rice thermo-sensitive male sterile lines, we knocked out the TMS5 genes of six elite japonica and four indica rice varieties by using CRISPR/Cas9 gene editing technology. By analyzing the critical sterility-inducing temperature (CIST) of the newly TGMS lines, it was found that the CIST of japonica TGMS lines ZG75S, CYGS, YG0618S, ZG07S, T0361S, and 7679S were between 28°C and 32°C, the CIST of indica TGMS lines 2537S, 6150S and 6379S were between 24°C and 28°C, and the CIST of indica TGMS line 1109S was lower than 23.5°C. These results indicated that the CIST of tms5 mutant from different genetic background materials was different. The TGMS lines with lower CIST could be obtained by knocking out the TMS5 from different genetic background materials. A hybrid rice combination 1109S / 8048 had high quality and high yield. The yield of 1109S/8048 was 13.1% higher than that of Fengliangyou 4. The creation of the TGMS 1109S and the high-yield cross combination 1109S/8048 provides a new way for high-yield breeding.

Introduction Colorectal cancer (CRC) is one of the most common illnesses that seriously threatens human health; many papers have reported that microRNAs (miRNAs) are promising biomarkers for cancer detection. However, miRNAs have not been used in clinical practice even though they are superior to the currently used screening tools, such as the fecal occult blood test (FOBT) and carcinoembryonic antigen (CEA) measurement. Methods In this study, we focused on the usefulness of a panel of miRNAs and the combination of miRNAs with the FOBT and CEA measurement, the currently used general diagnosis methods, to improve the accuracy of CRC diagnosis. Results The results showed that the miRNA panel has great potential value as a diagnostic biomarker with high specificity and sensitivity, and further analysis demonstrated that the miRNA panel had higher sensitivity and specificity than the FOBT and CEA measurement, even when these methods were combined. More importantly, although the miRNA panel is superior to the FOBT and CEA measurement, it cannot replace them. Conclusions In this research, we investigated whether complementarity exists between the miRNA panel and the FOBT and CEA measurement for CRC diagnosis. Interestingly, the results indicated that the FOBT and CEA measurement could improve the positivity rate of the miRNA panel as a biomarker and vice versa.

The genus Cuneopsis Simpson, 1900 comprises seven valid species, and Cuneopsis celtiformis (Heude, 1874) is the type species of this genus. Previous phylogenetic studies using complete mitochondrial genomes showed that Cuneopsis was not monophyletic, but the result was hampered by incomplete species sampling and lack of the type species of this genus. In this study, we collected C. celtiformis from the type locality and determined its complete maternal mitochondrial genome. This mitogenome is 15,922 bp in length and contains 14 protein-coding genes (including one F-orf), two rRNA genes, 22 tRNA genes, and 1 putative control region. Our mitochondrial phylogenomic analysis confirms that currently recognized genus Cuneopsis is polyphyletic, and C. celtiformis is the closest to C. heudei with high maximum likelihood bootstrap support value. Comprehensive sampling of all Cuneopsis species is needed for phylogenetic analysis to erect new genera in future studies.

Objective To generate a comprehensive transcriptomic profile of cellular signifures and transcriptomes of human placenta in GDM by single-cell RNA-sequencing (scRNA-seq), and built a comprehensive cell atlas. We hope to reveal the molecular mechanism of pregnancy risk for GDM women. Design Scientific study. Setting University Hospital and laboratories. Population or Sample Pregnant women Methods 20 GDM women and 20 normal pregnant women were recruited in present study. ScRNA-seq were loaded on the Chromium Single Cell Controller Instrument (10×Genomics) to generate single cell gel beads in emulsions (GEMs). Results A total of 27,220 cells from placenta samples were obtained. First, with the cell-type-specific marker genes, we annotated 15 cell clusters into more than 9 different cell types. Second, beside the classcial markers, we also found some novel markers for distinguishing three kinds of trophoblast and subtypes, which cloud be confirmed by immunohistochemistry imaging. Third, we demonstrated the specific placental function in GDM by the bioinformatics analysis of differentially expressed genes, such as estrogen signaling pathway, natural killer cell mediated cytotoxicity down- regulated. Fourth, there are abundant ligand-receptor interactions between trophoblast and immune cells in the maternal fetal interface microenvironment, such as VEGFB-FLT1, MIF-EGFR, RPS19-C5AR1, SPP1-CD44These dysfunctional ligand-receptor interactions may play the important roles in the development of GDM. Conclusion This study provided the first cell-type-specific transcriptomic alterations GDM placenta from single cell level, exploded the cell identities and cell-type-specific marker genes in the human placenta. In addition, it demonstrated the features of placental function and cell interactions for GDM.

Canonical epigenetic modifications, which include histone modification, chromatin remodeling and DNA methylation, play key roles in numerous cellular processes. Epigenetics underlies how cells that posses DNA with similar sequences develop into different cell types with different functions in an organism. Earlier epigenetic research has primarily been focused at the chromatin level. However, the number of studies on epigenetic modifications of RNA, such as N1-methyladenosine, 2′-O-ribosemethylation, inosine, 5-methylcytidine, N6-methyladenosine (m6A) and pseudouridine, has seen an increase. Circular RNAs (circRNAs), a type of RNA species that lacks a 5′ cap or 3′ poly(A) tail, are abundantly expressed in acute myeloid leukemia (AML) and may regulate disease progression. circRNAs possess various functions, including microRNA sponging, gene transcription regulation and RNA-binding protein interaction. Furthermore, circRNAs are m6A methylated in other types of cancer, such as colorectal and hypopharyngeal squamous cell cancers. Therefore, the critical roles of circRNA epigenetic modifications, particularly m6A, and their possible involvement in AML are discussed in the present review. Epigenetic modification of circRNAs may become a diagnostic and therapeutic target for AML in the future.

This study explored the influence of biochar (BC) on anaerobic digestion (AD) of swine manure under various tetracycline (TC) pressures. It was found that both low (0.5 mg/L) and high (50 mg/L) TC pressures inhibited AD performance, while BC mitigated it in multi-facets. Under high TC pressure, BC accelerated syntrophic methanogenesis by boosting direct interspecies electron transfer pathway. The TC removal efficiencies were enhanced by 24.3-158.2% with BC assistance, which were attributed to the enhanced biological degradation rather than BC’s physiochemical adsorption. Moreover, BC possibly acted as a protective role to alleviate intensive extracellular polymeric substances secretion under TC pressures. Integrated microbial community, metabolic function predicting, and antibiotic resistance genes (ARG) analysis revealed that BC addition not only enriched Anaerolineceae, which likely responsible for the 24.2-41.9% higher level expression of organics metabolic pathways and xenobiotics biodegradation, but also reduced ARG abundance by controlling the potential ARG host (Firmicutes) proliferation.

To clarify the roles of biochar in the anaerobic co-digestion of waste activated sludge (WAS) and food waste (FW), batch tests were conducted coupled with thermodynamics, extracellular electron transfer flux and microbial community analysis. Compared with the control group, biochar significantly facilitated the co-digestion at three periods, but its sustainable facilitation was mainly in the syntrophic methanogenesis of volatile fatty acids (VFAs). The thermodynamic analysis confirmed that biochar could alleviate limitations imposed by high hydrogen partial pressure during interspecies hydrogen transfer (IHT), the thermodynamic windows was expanded 137% and 92% in the syntrophic methanogenesis of acetate and propionate, respectively. Meanwhile, due to the redox capacity of biochar (4.85 and 0.35 μmol e−/g biochar), the equivalent current of direct interspecies electron transfer (DIET) flux for syntrophic methanogenesis of acetate and propionate obtained were 1.0 × 10−4 A and 0.9 × 10−4 A, which were 108 times than that of IHT. It should be noticed that the functional microorganisms like Methanosarcina which could participate DIET were only enriched on the surface of biochar, the dominant Methanothermobacter in suspended sludge probably indicate IHT was still the main pathway for syntrophic methanogenesis. Nevertheless, the DIET triggered by the redox-active moieties on the surface of biochar and the enhanced IHT by alleviating thermodynamic restrictions, promoted the syntrophic methanogenesis synergistically.

Frameshift mutations are generally considered to be lethal because it could result in radical changes of the protein sequence behind. However, the protein of frameshift mutants of a type I toxin (ibsc) was found to be still toxic to bacteria, retaining the similar function as wild-type protein to arrest the cellular growth by impairing the membrane's integrity. Additionally, we have verified that this observation is not an individual event as the same phenomenon had been found in other toxins subsequently. After analyzing the coding sequence of these genes, we proposed a hypothesis to search this kind of hidden gene, through which a dihydrofolate reductase-encoding gene (dfrB3) was found out. Like the wild-type reductase, both +1 and –1 frame-shifted proteins of dfrB3 gene were also proved to catalyze the reduction of dihydrofolate to tetrahydrofolate by using NADPH.

Dysregulation of early neurodevelopment is implicated in macrocephaly/autism disorders. However, the mechanism underlying this dysregulation, particularly in human cells, remains poorly understood. Mutations in the small GTPase gene RAB39b are associated with X-linked macrocephaly, autism spectrum disorder (ASD), and intellectual disability. The in vivo roles of RAB39b in the brain remain unknown. We generated Rab39b knockout (KO) mice and found that they exhibited cortical neurogenesis impairment, macrocephaly, and hallmark ASD behaviors, which resembled patient phenotypes. We also produced mutant human cerebral organoids that were substantially enlarged due to the overproliferation and impaired differentiation of neural progenitor cells (NPCs), which resemble neurodevelopmental deficits in KO mice. Mechanistic studies reveal that RAB39b interacts with PI3K components and its deletion promotes PI3K–AKT–mTOR signaling in NPCs of mouse cortex and cerebral organoids. The mTOR activity is robustly enhanced in mutant outer radial glia cells (oRGs), a subtype of NPCs barely detectable in rodents but abundant in human brains. Inhibition of AKT signaling rescued enlarged organoid sizes and NPC overproliferation caused by RAB39b mutations. Therefore, RAB39b mutation promotes PI3K–AKT–mTOR activity and alters cortical neurogenesis, leading to macrocephaly and autistic-like behaviors. Our studies provide new insights into neurodevelopmental dysregulation and common pathways associated with ASD across species.

We report on a supramolecular sensor array using fluorogenic peptide probes and graphene oxide that can target glycoproteins on a viral caspid, facilitating the differentiation of ebola virus from marburg virus and receptor-extensive vesicular stomatitis virus using principal component analysis.

Angiogenesis enhances cancer metastasis and progression, however, the roles of transcription regulation in angiogenesis are not fully defined. ZNF322A is an oncogenic zinc-finger transcription factor. Here, we demonstrate a new mechanism of Kras mutation-driven ZNF322A transcriptional activation and elucidate the interplay between ZNF322A and its upstream transcriptional regulators and downstream transcriptional targets in promoting neo-angiogenesis. Methods: Luciferase activity, RT-qPCR and ChIP-qPCR assays were used to examine transcription regulation in cell models. In vitro and in vivo angiogenesis assays were conducted. Immunohistochemistry, Kaplan-Meier method and multivariate Cox regression assays were performed to examine the clinical correlation in tumor specimens from lung cancer patients. Results: We validated that Yin Yang 1 (YY1) upregulated ZNF322A expression through targeting its promoter in the context of Kras mutation. Reconstitution experiments by knocking down YY1 under KrasG13V activation decreased KrasG13V-promoted cancer cell migration, proliferation and ZNF322A promoter activity. Knockdown of YY1 or ZNF322A attenuated angiogenesis in vitro and in vivo. Notably, we validated that ZNF322A upregulated the expression of sonic hedgehog (Shh) gene which encodes a secreted factor that activates pro-angiogenic responses in endothelial cells. Clinically, ZNF322A protein expression positively correlated with Shh and CD31, an endothelial cell marker, in 133 lung cancer patient samples determined using immunohistochemistry analysis. Notably, patients with concordantly high expression of ZNF322A, Shh and CD31 correlated with poor prognosis. Conclusions: These findings highlight the mechanism by which dysregulation of Kras/YY1/ZNF322/Shh transcriptional axis enhances neo-angiogenesis and cancer progression in lung cancer. Therapeutic strategies that target Kras/YY1/ZNF322A/Shh signaling axis may provide new insight on targeted therapy for lung cancer patients.

Steric hindrance in the binding pocket of an alcohol dehydrogenase (ADH) has a great impact on its activity and stereoselectivity simultaneously. Due to the subtle structural difference between two bulky phenyl substituents, the asymmetric synthesis of diaryl alcohols by bioreduction of diaryl ketones is often hindered by the low activity and stereoselectivity of ADHs. To engineer an ADH with practical properties and to investigate the molecular mechanism behind the asymmetric biocatalysis of diaryl ketones, we engineered an ADH from Lactobacillus kefiri (LkADH) to asymmetrically catalyse the reduction of 4-chlorodiphenylketones (CPPK), which are not catalysed by the wild type (WT) enzyme. Mutants seq1–seq5 with gradually increased activity and stereoselectivity were obtained through iterative “shrinking mutagenesis.” The final mutant seq5 (Y190P/I144V/L199V/E145C/M206F) demonstrated the highest activity and excellent stereoselectivity of >99% ee. Molecular simulation analyses revealed that mutations may enhance the activity by eliminating steric hindrance, inducing a more open binding loop and constructing more noncovalent interactions. The pro-R pose of CPPK with a halogen bond formed a pre-reaction conformation more easily than the pro-S pose, resulting in the high ee of (R)-CPPO in seq5. Moreover, different halogen bonds formed due to the different positions of chlorine substituents, resulting in opposite substrate binding orientation and stereoselectivity. Therefore, the stereoselectivity of seq5 was inverted toward ortho- rather than para-chlorine substituted ketones. These results indicate that the stereocontrol element of LkADH was changed to recognise diaryl ketones after steric hindrance was eliminated. This study provides novel insights into the role of steric hindrance and noncovalent bonds in the determination of the activity and stereoselectivity of enzymes, and presents an approach producing key intermediates of chiral drugs with practical potential.