Increased gene dosage of RFWD2 causes autistic-like behaviors and aberrant synaptic formation and function in mice.

Autism spectrum disorder (ASD) is a neurodevelopmental disorder characterized by impaired social interactions, communication deficits and repetitive behaviors. A study of autistic human subjects has identified RFWD2 as a susceptibility gene for autism, and autistic patients have 3 copies of the RFWD2 gene. The role of RFWD2 as an E3 ligase in neuronal functions, and its contribution to the pathophysiology of ASD, remain unknown. We generated RFWD2 knockin mice to model the human autistic condition of high gene dosage of RFWD2. We found that heterozygous knockin (Rfwd2+/-) male mice exhibited the core symptoms of autism. Rfwd2+/- male mice showed deficits in social interaction and communication, increased repetitive and anxiety-like behavior, and spatial memory deficits, whereas Rfwd2+/- female mice showed subtle deficits in social communication and spatial memory but were normal in anxiety-like, repetitive, and social behaviors. These autistic-like behaviors in males were accompanied by a reduction in dendritic spine density and abnormal synaptic function on layer II/III pyramidal neurons in the prelimbic area of the medial prefrontal cortex (mPFC), as well as decreased expression of synaptic proteins. Impaired social behaviors in Rfwd2+/- male mice were rescued by the expression of ETV5, one of the major substrates of RFWD2, in the mPFC. These findings indicate an important role of RFWD2 in the pathogenesis of autism.

Ternary XBe4H5- (X = Si, Ge, Sn, Pb) Clusters: Planar Tetracoordinate Si/Ge/Sn/Pb Species with 18 Valence Electrons.

As one of the important probes of chemical bonding, planar tetracoordinate carbon (ptC) compounds have been receiving much attention. Compared with ptC clusters, the heavier planar tetracoordinate silicon, germanium, tin, lead (ptSi/Ge/Sn/Pb) systems are scarcer and more exotic. The 18-valence-electron (ve)-counting is one important guide, though not the only rule, for the design of planar tetra-, penta-coordinate carbon and silicon clusters. The 18ve ptSi/Ge system is very scarce and needs to be expanded. Based on the isoelectronic principle and bonding similarity between the Al atom and the BeH unit, inspired by the previously reported ptSi global minimum (GM) SiAl42-, a series of ternary 18 ve XBe4H5- (X = Si, Ge, Sn, Pb) clusters were predicted with the ptSi/Ge/Sn/Pb centers. Extensive density functional theory (DFT) global minimum searches and high-level CCSD(T) calculations performed herein indicated that these ptSi/Ge/Sn/Pb XBe4H5- (X = Si, Ge, Sn, Pb) clusters were all true GMs on their potential energy surfaces. These GMs of XBe4H5- (X = Si, Ge, Sn, Pb) species possessed the beautiful fan-shaped structures: XBe4 unit can be stabilized by three peripheries bridging H and two terminal H atoms. It should be noted that XBe4H5- (X = Si, Ge, Sn, Pb) were the first ternary 18 ve ptSi/Ge/Sn/Pb species. The natural bond orbital (NBO), canonical molecular orbitals (CMOs) and adaptive natural densitpartitioning (AdNDP) analyses indicated that 18ve are ideal for these ptX clusters: delocalized one π and three σ bonds for the XBe4 core, three Be-H-Be 3c-2e and two Be-H σ bonds for the periphery. Additionally, 2π plus 6σ double aromaticity was found to be crucial for the stability of the ptX XBe4H5- (X = Si, Ge, Sn, Pb) clusters. The simulated photoelectron spectra of XBe4H5- (X = Si, Ge, Sn, Pb) clusters will provide theoretical basis for further experimental characterization.

Development and validation of a nomogram for predicting survival in patients with acute pancreatitis.

BACKGROUND: Acute pancreatitis (AP) is a complex and heterogeneous disease. We aimed to design and validate a prognostic nomogram for improving the prediction of short-term survival in patients with AP. METHODS: The clinical data of 632 patients with AP were obtained from the Medical Information Mart for Intensive Care (MIMIC)-IV database. The nomogram for the prediction of 30-day, 60-day and 90-day survival was developed by incorporating the risk factors identified by multivariate Cox analyses. RESULTS: Multivariate Cox proportional hazard model analysis showed that age (hazard ratio [HR]=1.06, 95% confidence interval [95% CI] 1.03-1.08, P<0.001), white blood cell count (HR=1.03, 95% CI 1.00-1.06, P=0.046), systolic blood pressure (HR=0.99, 95% CI 0.97-1.00, P=0.015), serum lactate level (HR=1.10, 95% CI 1.01-1.20, P=0.023), and Simplified Acute Physiology Score II (HR=1.04, 95% CI 1.02-1.06, P<0.001) were independent predictors of 90-day mortality in patients with AP. A prognostic nomogram model for 30-day, 60-day, and 90-day survival based on these variables was built. Receiver operating characteristic (ROC) curve analysis demonstrated that the nomogram had good accuracy for predicting 30-day, 60-day, and 90-day survival (area under the ROC curve: 0.796, 0.812, and 0.854, respectively; bootstrap-corrected C-index value: 0.782, 0.799, and 0.846, respectively). CONCLUSION: The nomogram-based prognostic model was able to accurately predict 30-day, 60-day, and 90-day survival outcomes and thus may be of value for risk stratification and clinical decision-making for critically ill patients with AP.

New Frontier in Antiviral Drugs for Disorders of the Respiratory System.

BACKGROUND: COVID-19 is still soaring, and the new delta COVID-19 variant is on the rise and spreading around the world. OBJECTIVE: We conducted a patent analysis to better understand the therapeutic strategy developed for antivirals available for the disorders of the respiratory system. MATERIALS AND METHODS: European granted patents filed from January 2002 to June 2021 were analyzed. We used a combination of International patent classification (IPC) "A61p31/12" and "A61p11/00" to search the relevant documents. RESULTS: Our study showed R&D of antiviral drugs for disorders of the respiratory system to be decreasing over the past 20 years. Chemical drugs showed various chemical structures. The development of chemical drugs or herbal medicines appeared to commence earlier than the biological products. Also, the results indicated that large global companies play a leading role in developing kinase inhibitors as chemical drugs. CONCLUSION: There are three strategies for developing antiviral drugs for the disorders of the respiratory system, including chemical drugs, herbal medicines or natural products, and biological products. Herbal medicines may provide a new insight and approach to developing antiviral drugs for disorders of the respiratory system. A combination of chemical drugs and natural products may be a promising therapeutic method for treating patients with COVID- 19.

Neutrophil-to-Lymphocyte Ratio (NLR) and Monocyte-to-Lymphocyte Ratio (MLR) Predict Clinical Outcome in Patients with Stage IIB Cervical Cancer.

INTRODUCTION: Stage IIB cervical cancer (CC) is an advanced stage CC with poor prognosis. Inflammatory response plays a crucial role in the development of CC, and systemic inflammatory indexes were related to the prognosis in several cancers. The objective of the study was to determine the prognostic value of platelet-to-lymphocyte ratio (PLR), neutrophil-to-lymphocyte ratio (NLR), monocyte-to-lymphocyte ratio (MLR), basophil-to-lymphocyte ratio (BLR), and systemic inflammation response index (SIRI) as inflammatory indexes in patients with stage IIB CC. MATERIALS AND METHODS: A retrospective study was performed in 260 patients with stage IIB CC. PLR, NLR, MLR, BLR, and SIRI were obtained from routine blood tests. Prognosis information of the patients was acquired from regular clinical follow-up. Recurrence and response to therapy were determined through electronic medical records (EMRs). Correlations of the inflammatory indexes with overall survival (OS), progression-free survival (PFS), recurrence, and response to therapy were analyzed using SPSS version 26.0 software. RESULTS: Receiver operating characteristic (ROC) curve analyses suggested that NLR, MLR, and SIRI had better predictive value than PLR as well as BLR in the prognosis and recurrence risk. Both univariate and multivariate survival analyses showed that higher NLR and MLR were significantly associated with shorter OS as well as PFS, whereas SIRI was not an independent predictive factor of PFS. Chi-square test results revealed that increased NLR was significantly correlated with higher recurrence rate (P=0.046), and increased MLR showed significant correlation with elevated recurrence risk (P=0.002). Univariate and binary logistic regression analyses for response to therapy indicated that elevated NLR was associated with decreased complete remission (CR) rate (P=0.031), and the P value lost statistical significance while being adjusted by tumor size (P=0.108). CONCLUSIONS: For patients with stage IIB CC, both NLR and MLR are independent prognostic factors as well as risk factors for recurrence; NLR serves as a potential marker for therapeutic response.

Development of a Real-Time TaqMan PCR Method for Absolute Quantification of the Biocontrol Agent Esteya vermicola.

Esteya vermicola has been used as an effective biocontrol agent for the management of the pinewood nematode, Bursaphelenchus xylophilus. Tools for monitoring the colonization and parasitism patterns of E. vermicola are required for the development of highly effective biocontrol strategies. Because the TaqMan PCR technique is effective for quantification of species in environmental samples, a real-time PCR-based methodology was developed for absolute quantification of E. vermicola via internal standard addition and extrapolation of DNA quantity to hyphal length. Primers and a probe for the 28S ribosomal RNA gene of E. vermicola were designed, and nested TaqMan real-time PCR-based quantification was performed. In addition, internal standard-based yield measurement was correlated to the absolute quantity of target genomic DNA. Moreover, an extrapolation curve obtained by optical microscopy and image analysis of the mycelia was constructed for the measurement of fungal hyphal length. The absolute quantification method developed in the present study provides a sensitive and accurate technique to quantify fungal density in either wood or other substrate samples and can be used as an effective tool for future studies of biocontrol agents.

In vivo infection of Bursaphelenchus xylophilus by the fungus Esteya vermicola.

BACKGROUND: As the causal agent of pine wilt disease, Bursaphelenchus xylophilus, is a serious pathogen of forest pine trees. Esteya vermicola is a nematophagous fungus of B. xylophilus and exhibits great potential as a biological control agent. However, the in vivo infection mechanism of E. vermicola on B. xylophilus is unclear. Experiments were conducted to study the colonization of host plant and infection of B. xylophilus by E. vermicola inside pine tree xylem. RESULTS: A green fluorescent protein (GFP)-tagged E. vermicola transformant was constructed as a biomarker to study the in vivo colonization and infection of B. xylophilus in pine trees. The in vitro infection of B. xylophilus by E. vermicola was observed through GFP expression. The bacilloid conidia produced by trophic hyphae in the body of the nematode are described. Additionally, the monitoring of in vivo colonization by GFP-tagged E. vermicola showed the germination and hyphal extension of this fungus after inoculation. Moreover, B. xylophilus infected by this biocontrol agent were extracted from healthy seedlings and observed in the xylem of trees that were wilting due to pine wilt disease. CONCLUSION: Evidence of fungal colonization and infection of B. xylophilus by E. vermicola is provided to improve our understanding of the in vivo infection mechanisms used by this nematophagous fungus against B. xylophilus. The infection of B. xylophilus by E. vermicola was inferred to begin with the implantation of propagules, and this inference will require future investigation. The colonization of Esteya vermicola in host pine tree xylem and the in vivo infection of pinewood nematode by E. vermicola were investigated using the green fluorescence protein transformant. © 2020 Society of Chemical Industry.

Salicylic acid in Populus tomentosa is a remote signalling molecule induced by Botryosphaeria dothidea infection.

The salicylic acid (SA) plays a critical role during the establishment of systemic acquired resistance (SAR) in uninfected plant tissues after localised exposure to a pathogen. Here, we studied SA in Populus tomentosa infected by the plant pathogen Botryosphaeria dothidea. The accumulation of SA and methyl salicylate (MeSA) occurred in chronological order in P. tomentosa. The SA and MeSA contents were greater at infected than uninfected sites. Additionally, a gene expression analysis indicated that SA might be accumulated by phenylalanine ammonialyase (PAL) and converted to MeSA by SA carboxyl methyltransferase (SAMT), while MeSA might convert to SA by SA-binding protein 2 (SABP2). The expressions of SAMT at infected sites and SABP2 at uninfected sites, respectively, were significantly up-regulated. Thus, SA might be converted to MeSA at infected sites and transported as a signalling molecule to uninfected sites, where it is converted to SA for SAR. Moreover, the expressions of pathogenesis-related genes PR-1, PR-2 and PR-5 in P. tomentosa were up-regulated by the B. dothidea infection. Our study determined that variations in SA and MeSA contents occur at infected and uninfected sites in poplar after pathogen infection and contributed to the remote signals for poplar SAR.

Glycogen Synthase Kinase-3 Modulates Hyperosmotic-Induced Urea Transporter A1 Relocation in the Inner Medullary Collecting Duct Cells.

AIM: Glycogen synthase kinase 3 (GSK3) regulates urine concentration by mediating the vasopressin-induced aquaporin 2 expression and water permeability, although it is unknown whether GSK3 also mediates the accumulation of the urea transporter A1 (UT-A1). The aim of this study is to investigate the effect of GSK3 on UT-A1 distribution. METHODS: Mouse inner medullary collecting duct 3 cells were transfected with UT-A1-GFP construct. The stable transfected cells were cultured under hypertonic conditions, treated with GSK3 inhibitor lithium chloride, GSK3 activator, lysosome or proteasome inhibitor. The expression levels of UT-A1, GSK3, and phospho-GSK3 were analyzed using western blot. The interaction between UT-A1 and the Golgi apparatus was examined using confocal immunofluorescence microscope. The UT-A1 trafficking was examined using the biotinylation of surface membranes. RESULTS: UT-A1 dissociated away from the Golgi apparatus and translocated to the plasma membrane under hypertonic-NaCl and NaCl plus urea stimulation. This movement was accompanied by the increased phosphorylation of GSK3 and its localization on the cellular membrane. Moreover, these results were duplicated by treating the cells with the GSK3 inhibitor, and by contrast, were partially reversed by the GSK3 activator. Treating cells with a lysosome or proteasome inhibitor failed to attenuate the effects of hypertonic stimulus, indicating that the loss of UT-A1 from the Golgi was not due to degradation. CONCLUSION: Our results suggest that GSK3 may in part modulate the hypertonic-induced intracellular UT-A1 redistribution and its accumulation on the plasma membrane, which may constitute another mechanism by which GSK3 modulates urine concentration.

[Pollution Characteristics and Health Risk Assessment of Polycyclic Aromatic Hydrocarbons in the Surface Soils of a Large Steel enterprise in the North of China].

16 priority control polycyclic aromatic hydrocarbons (PAHs) were analyzed by GC-MS in 22 surface soil (0-20 cm) samples collected from a large steel enterprise in the north of China. The concentrations of Σ16PAHs ranged from 22.0 µg·kg-1 to 20062.0 µg·kg-1. 4 to 5 aromatic rings were the dominant, typically fluorene(Flu) and pyrene(Pyr). Compared with related domestic research, PAH pollution in the steel enterprise reached medium level, with fifty percent of the soil points were at moderate and severe pollution levels, mainly in the coking and pellet plant area. The concentrations of 10 PAHs in 20 soil samples exceeded the Dutch target reference values. Compared with soil screening value of Beijing contaminated industrial sites, only part of the sample points exceeded the standard, typically benzo[a]anthracene(BaA) and benzo[a]pyrene(BaP). The source apportionment showed that soil PAHs mainly originated from combustion products of coal and other fossil fuels with only a small portion contributed by oil combustion and spill. The health risk assessment showed that the carcinogenic risks of benzo[a]pyrene(BaP), benzo[a]anthracene(BaA), dibenz(a,h)anthracene(DBA), benzo[b]fluoranthene(BbF), indeno[1,2,3-cd]pyrene(InP) exceeded the threshold of 1×10-6 under residential land condition, the carcinogenic risks of benzo[a]pyrene(BaP), benzo[a]anthracene(BaA), dibenz(a,h)anthracene(DBA) also exceeded the threshold of 1×10-6 under industrial land condition. The carcinogenic risk value of benzo[a]pyrene(BaP) was the biggest among the 16 PAHs. The soil PAHs in the steel enterprise already caused harm to human health and the soil restoration project must be carried out.

[Speciation and Risk Assessment of Heavy Metals in Surface Sediments from the Heavily Polluted Area of Xiaoqing River].

Th concentrations of Cu, As, Pb, Zn, Cr, Cd, Ni in interstitial water were analyzed by ICP-MS from the heavily polluted area of Xiaoqing River. A modified BCR's sequential extraction procedure was used to investigate the fraction of the heavy metals in the surface sediments. The aquatic toxicity of heavy metals in interstitial water was assessed by US Water Quality Criteria (CCC, CMC). Based on the speciation of the metals in the surface sediments, the ecological risk of heavy metals was assessed with methods of risk assessment code (RAC) and potential ecological risk index. The results showed that: The tested heavy metals would not pose chronic toxicity for aquatic ecosystem. The contents of heavy metals (Cu, Pb, Zn, Cr, Cd, Ni) in the surface sediments were higher than the background values of the local soils, indicating enrichment of heavy metals. Cu, As and Ni were mainly composed with residual fractions, Pb and Cr were mainly constituted of residual and oxidizable fractions, and mass fractions of Zn and Cd existed mainly in acid soluble and reducible factions. The contents of bio-available fractions of Cd, Zn, Cr and Pb in the sediments were higher those of residual fractions, indicating high potential for secondary release. Based on calculation of RAC method, Cu, Pb, Cr and As posed extremely low to low risk to the environment. Accordingly, Ni was of low to medium risk, Cd was of medium to high risk, and Zn was of medium to extremely high risk to the environment. The potential ecological risk of heavy metals in the sediments was in the descending order of Cd > Zn > Ni > As > Cu > Cr > Pb. Cd had high to very high potential ecological risk comparing with the other heavy metals. The potential ecological risk indexes (RI) of the heavy metals in the sediments were in the range of 136.83-264.83, and posed medium to high potential ecological risks for Xiaoqing River.

Cytospora species from Populus and Salix in China with C. davidiana sp. nov.

Poplar and willow plantations have become widespread in China, in order to meet national economic and environmental needs. The emergence of several pathogens is enhanced by climatic change and associated human factors. Species of Cytospora are well-known pathogens on poplar and willow, and cause stem cankers and diebacks. In the present study, we conducted a survey of Cytospora species occurring on Populus spp. and Salix spp. in China. We used morphological examination and phylogenetic inferences, based on the DNA sequence data from the internal transcribed spacer regions (ITS1, 5.8S rDNA, and ITS2) and partial ß-tubulin gene, to identify six Cytospora species occurring on poplar and willow. Five of these species belonged to known taxa, viz. Cytospora chrysosperma (asexual state of Valsa sordida), Cytospora translucens (asexual state of Leucostoma translucens), Cytospora fugax (asexual state of Valsa salicina), Cytospora atrocirrhata, and Cytospora kantschavelii. Our study yielded a new species, Cytospora davidiana sp. nov., on poplar. The new species is characterized by typical torsellioid conidiomata. An additional Cytospora sp. 1, which formed a distinct clade in the phylogenetic inferences, remains unnamed; the paucity of available materials prevented phenotypical characterization.

Ad-endostatin treatment combined with low-dose irradiation in a murine lung cancer model.

Radiation therapy is a conventional strategy for treating advanced lung cancer yet is accompanied by serious side-effects. Its combination with other strategies, such as antiangiogenesis and gene therapy, has shown excellent prospects. As one of the potent endogenous vascular inhibitors, endostatin has been widely used in the antiangiogenic gene therapy of tumors. In the present study, LL/2 cells were infected with a recombinant adenovirus encoding endostatin (Ad-endostatin) to express endostatin. The results showed that LL/2 cells infected with the Ad-endostatin efficiently and longlastingly expressed endostatin. In order to further explore the role of Ad-endostatin combined with irradiation in the treatment of cancer, a murine lung cancer model was established and treated with Ad-endostatin combined with low-dose irradiation. The results showed that the combination treatment markedly inhibited tumor growth and metastasis, and prolonged the survival time of the tumor-bearing mice. Furthermore, this significant antitumor activity was associated with lower levels of microvessel density and anoxia factors in the Ad-Endo combined with irradiation group, and with an increased apoptotic index of tumor cells. In addition, no serious side-effects were noted in the combination group. Based on our findings, Ad-endostatin combined with low-dose irradiation may be a rational alternative treatment for lung cancer and other solid tumors.

Eryngiolide A, a cytotoxic macrocyclic diterpenoid with an unusual cyclododecane core skeleton produced by the edible mushroom Pleurotus eryngii.

Eryngiolide A (1), the first member of C20 diterpenoids with the skeleton deriving from a cyclododecane core fused with two γ-lactone units, was isolated from the solid culture of the edible mushroom Pleurotus eryngii. The structure of 1 was elucidated by extensive analysis of NMR spectra. Compound 1 exhibited moderate cytotoxicity against two human cancer lines in vitro.

AAV-mediated gene transfer of human pigment epithelium-derived factor inhibits Lewis lung carcinoma growth in mice.

Pigment epithelium-derived factor (PEDF) is the most potent inhibitor of angiogenesis in the mammalian eye, and mechanisms through which PEDF exerts its antitumour activity have recently been defined. The aim of our research was to evaluate the ability of adeno-associated virus (AAV) vector-mediated transfer of human PEDF to inhibit lewis lung carcinoma (LCC) cell growth. Intratumoural injection of AAV-PEDF caused significant reduction of the tumour volume and prolonged the survival time of mice bearing LLC cells, which were associated with decreased microvessel density and increased apoptosis in the tumours. AAV vectors represent a very promising tool for cancer gene therapy. No noticeable toxicity concerning AAV was detected as inferred from monitoring changes in animal body weight as well as basic organ structure and histological morphology, and by analyzing mouse liver and kidney function. Our findings indicate that AAV-mediated PEDF gene expression may offer an active approach to inhibit LLC growth and that treatment with AAV-PEDF may provide a promising therapeutic strategy in lung cancer treatment.