Small molecule agonist of mitochondrial fusion repairs mitochondrial dysfunction.

Membrane dynamics are important to the integrity and function of mitochondria. Defective mitochondrial fusion underlies the pathogenesis of multiple diseases. The ability to target fusion highlights the potential to fight life-threatening conditions. Here we report a small molecule agonist, S89, that specifically promotes mitochondrial fusion by targeting endogenous MFN1. S89 interacts directly with a loop region in the helix bundle 2 domain of MFN1 to stimulate GTP hydrolysis and vesicle fusion. GTP loading or competition by S89 dislodges the loop from the GTPase domain and unlocks the molecule. S89 restores mitochondrial and cellular defects caused by mitochondrial DNA mutations, oxidative stress inducer paraquat, ferroptosis inducer RSL3 or CMT2A-causing mutations by boosting endogenous MFN1. Strikingly, S89 effectively eliminates ischemia/reperfusion (I/R)-induced mitochondrial damage and protects mouse heart from I/R injury. These results reveal the priming mechanism for MFNs and provide a therapeutic strategy for mitochondrial diseases when additional mitochondrial fusion is beneficial.

MODILM: towards better complex diseases classification using a novel multi-omics data integration learning model.

BACKGROUND: Accurately classifying complex diseases is crucial for diagnosis and personalized treatment. Integrating multi-omics data has been demonstrated to enhance the accuracy of analyzing and classifying complex diseases. This can be attributed to the highly correlated nature of the data with various diseases, as well as the comprehensive and complementary information it provides. However, integrating multi-omics data for complex diseases is challenged by data characteristics such as high imbalance, scale variation, heterogeneity, and noise interference. These challenges further emphasize the importance of developing effective methods for multi-omics data integration. RESULTS: We proposed a novel multi-omics data learning model called MODILM, which integrates multiple omics data to improve the classification accuracy of complex diseases by obtaining more significant and complementary information from different single-omics data. Our approach includes four key steps: 1) constructing a similarity network for each omics data using the cosine similarity measure, 2) leveraging Graph Attention Networks to learn sample-specific and intra-association features from similarity networks for single-omics data, 3) using Multilayer Perceptron networks to map learned features to a new feature space, thereby strengthening and extracting high-level omics-specific features, and 4) fusing these high-level features using a View Correlation Discovery Network to learn cross-omics features in the label space, which results in unique class-level distinctiveness for complex diseases. To demonstrate the effectiveness of MODILM, we conducted experiments on six benchmark datasets consisting of miRNA expression, mRNA, and DNA methylation data. Our results show that MODILM outperforms state-of-the-art methods, effectively improving the accuracy of complex disease classification. CONCLUSIONS: Our MODILM provides a more competitive way to extract and integrate important and complementary information from multiple omics data, providing a very promising tool for supporting decision-making for clinical diagnosis.

First report of anthracnose caused by Colletotrichum siamense on Illicium verum in China.

Aniseed (Illicium verum) is a woody spice tree that has been grown in China for a long time. Anthracnose is an important disease of aniseed, which can cause severe leaf drop. In Sep. 2020, severe anthracnose was observed in Shanglin (23°35'5"N, 108°19'51"E), Nanning, Guangxi in China, and the incidence was 85%. The symptoms at the early stage were small, round and watery, then became larger and gradually turned brown. The acervuli would appear at the later stage, and contain many conidia. Leaves with disease were randomly sampled from 10 plants, and were cut into small rectangular pieces of 0.5×1 cm, and disinfected with 75% alcohol 1 min, with 0.1% HgCl2 3 min. After washing with sterile water 3 times, they were placed onto potato dextrose agar (PDA) medium and incubated at 25°C for 5 days. The average colony growth rate was 11.85 mm/d in 7 days. The colony was white or light gray in the initial stage, with dense aerial mycelium, and the central mycelium of the colony was dark grey in the later stage. Conidia were colorless, single spore, smooth, cylindrical, both ends obtuse, with an average size of 14.95 ± 0.97 µm × 5.46 ± 0.44 µm (n = 100). The conidial appressorium was oval or club-shaped, brown, margin intact, with an average size of 7.83 ± 1.21 µm × 5.82 ± 0.58µm (n = 50). Three strains GXNN02, GXNN03 and GXNN05 were selected for further study. Primer pairs T1/ßt2b, ACT512/ACT783, GDF/GDR, CHS1-79F / CHS1-354R and ITS1/ITS4 (Weir et al. 2012) were used to amplify tubulin (TUB), actin (ACT), 3-phosphate glyceraldehyde dehydrogenase (GAPDH), chitinase (CHS1) and the internal transcribed spacers of rDNA (ITS) respectively. BLASTn searches showed that the TUB (ON619861-63 ), ACT (ON619852-54), GAPDH (ON619855-57), CHS1 (ON619858-60) and ITS (ON573028-30) sequences had the highest similarity to Colletotrichum siamense with up to 99% (699/702, 676/679, 699/702) identity for TUB (JX010404.1); 99% (281/282, 253/254, 249/250) identity for ACT (JX009518.1); 99% (275/277, 275/277, 239/241) identity for GAPDH (JX009924.1); 99% (296/299, 296/299, 259/262) identity for CHS1 (JX009865.1); up to 99% (527/530, 485/487, 527/530) identity for ITS (JX010171.1) of ex-type ICMP 18578. A ML tree was constructed by combining 5 sequenced loci, and three isolates clustered in the C. siamense clade with 94% bootstrap support. Therefore, combined with the morphological characteristics, the pathogens were identified as C. siamense. In a pathogenicity test, these three isolates were tested on 9 healthy aniseed seedlings with at least 10 leaves, and 3 seedlings as control. The leaves were surface disinfected with 75% alcohol, and then wiped with sterilized water three times. Holes were made near the edge of the leaves and were sprayed with conidial solution (6×106 spores/mL) in test groups, and use sterilized water as control. Then the leaves were sealed inside a plastic bag for 48 h to retain moisture. Brown spot and black acervuli, similar to the symptoms in the field, were observed on the leaves in test groups within 10-15 days. No symptoms were observed on the negative control leaves. The pathogens were reisolated from the treated infected leaves and were identified as C. siamense, thus fulfilling Koch's postulates. The pathogenicity test was confirmed by repeating in triplicate. The isolation frequency of C. siamense in our samples was 82.50%. To our knowledge, this is the first report of C. siamense in China. Further research on the occurrence of the disease will help prevent the spread of the disease.

First report of leaf blight on Eucalyptus cloeziana caused by Coniella quercicola in China.

Eucalypt species are among the most important for timber production worldwide. Eucalyptus cloeziana is increasingly culticated due to its desirable structural properties. Leaf blight is one of the most devastating diseases of E. cloeziana in China. In May 2019, leaf blight samples were collected from E. cloeziana in Chongzuo, Guangxi, China (22°20'37.70"N, 107°49'29.29"E). Lesions began at the leaf margin and extended to 1/4-3/4 of the total leaf surface area. Lesions (26.76±12.64 mm diameter) were round, yellow, and withered in appearance, and sometimes many black, round pycnidia were observed. Leaves with blight were collected randomly from 10 E. cloeziana plants. Tissue blocks (3 mm×3 mm) were sampled from diseased and healthy leaf portions, then surface disinfected with 75% ethanol for 20 s and 0.1% HgCl2 for 3 min. After washing with sterile water three times, dry tissue blocks were placed on potato dextrose agar (PDA) medium and incubated at 28°C for 5 days. Hyphae were milky white or whitish, and sparse. The colonies had petal-shaped edges and the conidiophores were clustered, branched and transparent. Spore-forming cells were solitary and smooth; conidia were smooth, fusiform or oblong, transparent, blunt-based, mostly erect, and 16.54±2.19 × 3.38±0.77 µm (n=100 in each isolate) in size. Three representative isolates (AB-6, AB-9, AB-16) were selected for further study. For molecular identification, the internal transcribed spacer (ITS) region of rDNA, translation elongation factor 1-α (TEF1), and large subunit ribosomal RNA (LSU) were amplified with primers ITS1/ITS4 (White et al. 1990), EF1-983F/EF1-1567R (Rehner and Buckleyet al. 2005), and LR0R/LR5 (Vilgalys and Hesteret al. 1990), respectively. BLASTn searches showed that the ITS (OM280456, ON026088-89), TEF1 (ON055278-80) and LSU (OM281346, ON026097-98) sequences had the highest similarity to Coniella quercicola strains with: 99% (600/605, 600/605, 600/604) identity for ITS (MH859478.1); 98% (326/333, 327/334, 325/332) identity for TEF1 (KX833698.1); 99% (870/872, 833/834, 830/831) identity for LSU (MH871258.1) of ex-type CBS 904.69. A Neighbor-Joining phylogenetic tree was constructed by combining 3 sequenced loci. Three isolates clustered in the C. quercicola clade with 100% bootstrap support. Thus, based on morphological (Maas et al. 1979; Wang and Lin et al. 2004) and molecular characteristics, the pathogen was identified as C. quercicola. In a pathogenicity test, 20 healthy E. cloeziana seedlings with at least 5 leaves were divided into 4 groups: groups 1-3 were used to inoculate three isolates respectively, and the fourth group acted as control. After surface disinfection with 75% ethanol and wiping with sterile water, tiny wounds were maked made by inoculation needle on each leaf. Fungal culture plugsblocks cut from 3 isolates were placed on wounds in groups 1-3 respectively,. withWarter- agar blockplugs served as control in group 4. The leaves were covered with wet cotton and sealed in airtight bags to retain moisture at room temperature with natural light. After 3 days, light brown lesions were observed in groups 1-3, with no symptoms present in the control group. The pathogenicity test was confirmed by repeating in triplicate and fungi re-isolated from symptomatic leaves were identified as C. quercicola. To our knowledge, this is the first report of leaf blight on E. cloeziana caused by C. quercicola in China. This study increases our understanding of E. cloeziana leaf blight and future research may allow the development of targeted prevention methods for more effective disease controls.

CircUCP2 promotes the tumor progression of non-small cell lung cancer through the miR-149/UCP2 pathway.

Non-small cell lung cancer (NSCLC) is a highly lethal cancer, and better treatments are urgently needed. Many studies have implicated circular RNAs (circRNAs) in the progression of multiple malignant tumors. Nonetheless, the functions of circRNAs in NSCLC remain unclear. To study new targets for the treatment of NSCLC, circRNA expression profiling was performed on NSCLC tissues and para-carcinoma nonmalignant tissues. RNA was isolated and used for circRNA sequencing. Biological studies were performed in vitro and in vivo to determine the functions of circRNAs in NSCLC, including their functions in cell proliferation and migration. How circRNAs function in NSCLC was explored to clarify the underlying regulatory mechanisms. We found that circUCP2 was upregulated in NSCLC tissues compared with neighboring nonmalignant tissues. circUCP2 promoted the proliferation and metastasis of NSCLC cells. circUCP2 promoted NSCLC progression by sponging miR-149 and upregulating UCP2. The circUCP2/miR-149/UCP2 axis accelerates the progression of NSCLC, and circUCP2 may therefore be a novel diagnostic biomarker for the progression of NSCLC.

Passive margins in accreting Archaean archipelagos signal continental stability promoting early atmospheric oxygen rise.

Significant changes in tectonic style and climate occurred from the late Archaean to early Proterozoic when continental growth and emergence provided opportunities for photosynthetic life to proliferate by the initiation of the Great Oxidation Event (GOE). In this study, we report a Neoarchaean passive-margin-type sequence (2560-2500 million years ago) from the Precambrian basement of China that formed in an accretionary orogen. Tectonostratigraphic and detrital zircon analysis reveal that thermal subsidence on the backside of a recently amalgamated oceanic archipelago created a quiet, shallow water environment, marked by deposition of carbonates, shales, and shallow water sediments, likely hosts to early photosynthetic microbes. Distinct from the traditional understanding of passive margins generated by continental rifting, post-collisional subsidence of archipelago margins represents a novel stable niche, signalling initial continental maturity and foreshadowing great changes at the Archaean-Proterozoic boundary.

Alpine-style nappes thrust over ancient North China continental margin demonstrate large Archean horizontal plate motions.

Whether modern-style plate tectonics operated on early Earth is debated due to a paucity of definitive records of large-scale plate convergence, subduction, and collision in the Archean geological record. Archean Alpine-style sub-horizontal fold/thrust nappes in the Precambrian basement of China contain a Mariana-type subduction-initiation sequence of mid-ocean ridge basalt blocks in a 1600-kilometer-long mélange belt, overthrusting picritic-boninitic and island-arc tholeiite bearing nappes, in turn emplaced over a passive margin capping an ancient Archean continental fragment. Picrite-boninite and tholeiite units are 2698 ± 30 million years old marking the age of subduction initiation, with nappes emplaced over the passive margin at 2520 million years ago. Here, we show the life cycle of the subduction zone and ocean spanned circa 178 million years; conservative plate velocities of 2 centimeters per year yield a lateral transport distance of subducted oceanic crust of 3560 kilometers, providing direct positive evidence for horizontal plate tectonics in the Archean.

Serological Investigation and Genetic Characteristics of Pseudorabies Virus in Hunan Province of China From 2016 to 2020.

Pseudorabies (PR), caused by variant pseudorabies virus (PRV), is an economically important viral disease in China. Recently, PRV infection in humans has also received attention worldwide. To investigate the PRV infection in Hunan province, China, we collected a total of 18,138 serum specimens from 808 PRV-vaccinated pig farms cross this region during 2016-2020, and we detected the presence of PRV glycoprotein B (gB) and gE-specific antibodies. The enzyme-linked immunosorbent assay (ELISA) results revealed that 80.47% (14,596/18,138, 95 CI 79.9-81.0) and 23.55% (4,271/18,138, 95 CI 22.9-24.2) of serum samples were positive for PRV gB and gE-specific antibodies, respectively. Further analysis indicated that the seroprevalence of wild PRV infection was associated with the season and breeding scale (p < 0.01). In addition, five PRV strains were isolated from PRV-positive samples in Vero cells and the virus titers varied from 106.5 to 107.51 TCID50/0.1 ml. The phylogenetic analysis revealed that one isolate was a classical strain of PRV genotype II, and four other isolates belonged to the variants of genotype II. Collectively, the data indicate that the prevalence of PRV remains high in pigs in Hunan province, and the variant PRV strains are the major genotypes affecting the development of the pig industry.

Successful captive breeding of a Malayan pangolin population to the third filial generation.

Pangolins are threatened placental mammals distributed in Africa and Asia. Many efforts have been undertaken in the last century to maintain pangolins in captivity, but only a few of them succeeded in maintaining and keeping this species in a controlled environment. This study reports the first systematic breeding of the Critically Endangered Malayan pangolin (Manis javanica) in captivity. Our captive breeding approach successfully improved the reproductive rate for both wild and captive-born female pangolins. From 2016 to 2020, we had 33 wild pangolins and produced 49 captive-born offspring spanning three filial generations. The female offspring further bred 18 offspring, of which 14 (78%) were conceived during the first time of cohabitation with males, and four offspring were conceived during the second cohabitation event, suggesting that they may practice copulation-induced ovulation. We observed that captive-born female pangolins could reach sexual maturity at 7-9 months (n = 4), and male pangolins could mate and successfully fertilise females at nine months age (n = 1). We also observed a female pangolin conceiving on the eighth day after parturition (the fifth day after the death of its pup). Our captive pangolins had a female-biased sex ratio of 1:0.5 at birth, unlike other known captive-born mammals. Also, captive-born pangolins were generally more viable after successful weaning and had a similar gestation length (~185 days) to wild pangolins. Most importantly, we report the first self-sustaining captive population of Malayan pangolins, and this species has an efficient reproduction strategy. These advances provide more comprehensive information for people to understand pangolins, and have implications for conserving endangered Malayan pangolins and providing scientific guidance to the management of other pangolin species.

shRNA targeting PLK1 inhibits the proliferation and invasion of nasopharyngeal carcinoma cells.

BACKGROUND: Polo-like kinase 1 (PLK1) is a serine/threonine protein kinase, which has been studied as a potential gene therapy target for many years. PLK1 is overexpressed in a variety of tumors, and its expression often negatively correlated with patient prognosis. However, the role of PLK1 in nasopharyngeal carcinoma (NPC) is rarely studied. METHODS: Two recombinant vector plasmids were transfected into CNE2 cell lines by liposome transfection, CNE2/PLK1 shRNA target PLK1 mRNA, as well as a non-targeting control plasmid, CNE2/NC shRNA. Meanwhile, non-transfected cells (CNE2) were also used as controls. Real-time quantitative PCR (qRT-PCR) and Western blotting were performed to detect the transfection effect. The effects of the downregulation of PLK1 on cell biological behavior was evaluated in vitro by using CCK8, Transwell, colony-forming and flow-cytometry assays. RESULTS: PLK1 mRNA and protein were significantly inhibited in CNE2/PLK1 shRNA cells. Compared to control groups, the CNE2/PLK1 shRNA cells showed slower cell growth and a significantly decreased cell-cloning rate. Both migration and invasion were significantly inhibited in experimental cells. The proportions of G2-phase and apoptotic cells within the experimental group were significantly increased. CONCLUSIONS: Our results indicate that specific interference of PLK1 gene expression can significantly inhibit the proliferation and invasion of NPC (CNE2) cells.

Bioinformatic analysis of differentially expressed genes and identification of key genes in EBV-transformed lymphoblasts.

Although the Epstein-Barr virus (EBV) is a well-known human oncogenic virus, its molecular mechanisms involved in the transformation of healthy human cells remain poorly understood. In this study, human lymphocytes were isolated from the peripheral blood of healthy adults, and lymphocytes were transformed in vitro by EBV. Agilent human whole genome microarrays were used to detect the differential gene expression profiles of EBV-transformed lymphoblasts and healthy peripheral blood lymphocytes (PBLs). By constructing the gene functional network of EBV-induced lymphocyte transformation, we screened out candidate key genes in this process and verified their expression levels by real-time quantitative polymerase chain reaction (RT-qPCR) and Western blot. In the EBV-transformed lymphoblasts, 2335 differentially expressed genes, including 1328 up-regulated and 1007 down-regulated, were screened out. Five candidate key genes, namely, PLK1, E2F1, PTPN11, BIRC5 and FYN were mainly screened out according to the results of LIMMA, String, Cytoscape software analysis. RT-qPCR and Western blot showed that PLK1, E2F1, PTPN11, BIRC5 genes had increased expression levels, and FYN gene was down-regulated in EBV-transformed lymphoblasts. Silencing of PLK1 gene in Raji cells could inhibit cell proliferation and invasion, and induce cell cycle arrest and apoptosis. In conclusion, PLK1, E2F1, PTPN11, BIRC5 and FYN are the candidate key molecules of EBV-transformed lymphocytes.

Structural insights of human mitofusin-2 into mitochondrial fusion and CMT2A onset.

Mitofusin-2 (MFN2) is a dynamin-like GTPase that plays a central role in regulating mitochondrial fusion and cell metabolism. Mutations in MFN2 cause the neurodegenerative disease Charcot-Marie-Tooth type 2A (CMT2A). The molecular basis underlying the physiological and pathological relevance of MFN2 is unclear. Here, we present crystal structures of truncated human MFN2 in different nucleotide-loading states. Unlike other dynamin superfamily members including MFN1, MFN2 forms sustained dimers even after GTP hydrolysis via the GTPase domain (G) interface, which accounts for its high membrane-tethering efficiency. The biochemical discrepancy between human MFN2 and MFN1 largely derives from a primate-only single amino acid variance. MFN2 and MFN1 can form heterodimers via the G interface in a nucleotide-dependent manner. CMT2A-related mutations, mapping to different functional zones of MFN2, lead to changes in GTP hydrolysis and homo/hetero-association ability. Our study provides fundamental insight into how mitofusins mediate mitochondrial fusion and the ways their disruptions cause disease.

Comparative Transcriptomics of Buzura suppressaria (Lepidoptera: Geometridae) Assembled De Novo Yield Insights Into Response After Buzura suppressaria Nuclear Polyhedrosis Virus Infection.

Buzura suppressaria Guenee (Lepidoptera: Geometridae) is a defoliator that seriously harms eucalyptus trees in South China. Buzura suppressaria nuclear polyhedrosis virus (BsNPV) is a baculovirus that infects B. suppressaria with high specificity and efficiency. Transcriptomes of B. suppressaria were sequenced before and after BsNPV infection using an Illumina-based platform to probe for differentially expressed genes (DEGs) of B. suppressaria after viral infection. On average, ∼57.4 million high-quality clean reads were generated and assembled de novo into 69,761 unigenes. The NCBI nonredundant protein, Swiss-Prot, Kyoto Encyclopedia of Genes and Genomes (KEGG), Gene ontology (GO), and Cluster of Orthologous Groups databases were used to annotate unigenes through NCBI BLAST; 33,575 unigenes (48.1%) were then mapped to at least one of these databases, and 4,366 unigenes (6.3%) were mapped to all databases. Differential expression analysis showed that 25,212 unigenes were upregulated and 22,880 unigenes were downregulated in at least one pairwise comparison. Control versus 48 h had more DEGs than other two pairwise comparisons in either the GO or KEGG database, because the number of regulated gene would increase as BsNPV infected more tissues and would decrease as more tissues were disabled. To ascertain B. suppressaria immune response to BsNPV infection, DEGs were annotated to the GO and KEGG databases. In total, 89 GO categories are related to immune response and 1,007 DEGs are annotated to these GO categories. Furthermore, 7 downregulated DEGs and 37 upregulated were obtained simultaneously in all three groups. These DEGs were considered to possess a central role throughout viral infection.