Subcellular Colocalization Assay of Host Factors with Viral Replication Complex in the dsRNA Reporter Nicotiana benthamiana.

As obligate pathogens, plant viruses co-opt several host factors for viral replication. Double-stranded RNA (dsRNA) plays important roles in plants, including eliciting innate immune responses and RNA interference. dsRNA also represents the genetic entities of a number of viruses and is a marker of infection by positive-sense single-stranded RNA viruses. Previous detection methods for RNA viruses basically relied on immunological methods, but later researchers discovered that the dsRNA-binding domain of the Flock house virus B2 protein is a perfect alternative to the J2 mAb for sensitive and rapid detection of long dsRNA in vitro and in vivo, and developed B2:GFP transgenic Nicotiana benthamiana line. This method describes in detail how to visualize host factors in the viral replication complex in time and space with the help of B2:GFP transgenic plants, exemplified by Turnip mosaic virus (TuMV), a representative virus member of the Potyviruses.

Proteomics Identified UDP-Glycosyltransferase Family Members as Pro-Viral Factors for Turnip Mosaic Virus Infection in Nicotiana benthamiana.

Viruses encounter numerous host factors that facilitate or suppress viral infection. Although some host factors manipulated by viruses were uncovered, we have limited knowledge of the pathways hijacked to promote viral replication and activate host defense responses. Turnip mosaic virus (TuMV) is one of the most prevalent viral pathogens in many regions of the world. Here, we employed an isobaric tag for relative and absolute quantitation (iTRAQ)-based proteomics approach to characterize cellular protein changes in the early stages of infection of Nicotiana benthamiana by wild type and replication-defective TuMV. A total of 225 differentially accumulated proteins (DAPs) were identified (182 increased and 43 decreased). Bioinformatics analysis showed that a few biological pathways were associated with TuMV infection. Four upregulated DAPs belonging to uridine diphosphate-glycosyltransferase (UGT) family members were validated by their mRNA expression profiles and their effects on TuMV infection. NbUGT91C1 or NbUGT74F1 knockdown impaired TuMV replication and increased reactive oxygen species production, whereas overexpression of either promoted TuMV replication. Overall, this comparative proteomics analysis delineates the cellular protein changes during early TuMV infection and provides new insights into the role of UGTs in the context of plant viral infection.

Efficient Toughening of DGEBA with a Bio-Based Protocatechuic Acid Derivative.

In this work, a bio-based epoxy resin, protocatechuic acid diester epoxy resin, (PDEP), was synthesized using protocatechuic acid. The structure and properties of PDEP have been characterized by 1H NMR, 13C NMR, and Fourier transform infrared. After different contents of PDEP were added to diglycidyl ether of bisphenol A (DGEBA), the modified epoxy resins were cured by 4,4'-diaminodiphenylmethane (DDM). With the addition of a flexible long-chain bio-based monomer to improve toughness, the impact strength was 50 kJ·m-2 with only 5.0 wt % PDEP; compared with pure DGEBA, the impact strength was 27 kJ·m-2. Further, an increase in T g should be confirmed from the mechanical cross-linking density and rigidity group content. The single T g proved the homogeneous phase structure of the PDEP-cured resin. Morphology exhibiting the ductile fracture of the cured resin was confirmed by scanning electron microscopy. Overall, this work demonstrates the utilization of renewable protocatechuic acid as an effective modifier for epoxy resin, which reflects its potential application.

Involvement of regulation of the excitation:inhibition functional balance in the mPFC in the antidepressant-anxiolytic effect of YL-IPA08, a novel TSPO ligand.

TSPO, an 18 kDa translocator protein, has received increased attention due to its antidepressant-anxiolytic effects. The balance between glutamatergic and GABAergic (E: I) in the medial prefrontal cortex (mPFC) is crucial for antidepressant-anxiolytic effects. However, no evidence is available to clarify the relationship between TSPO and E:I balance. In the present study, we used the TSPO global-knockout (KO) and TSPO wild-type (WT) mice to assess the effects of TSPO on antidepressant-anxiolytic effects of YL-IPA08 (a novel TSPO ligand) and the underlying neurobiological mechanism. Additionally, a multichannel electrophysiological technique was used to explore the effects of YL-IPA08 on pyramidal neurons and interneurons in mPFC. Open field test (OFT) and elevated plus maze (EPM) test revealed that a single dose of YL-IPA08 (0.3 mg/kg, i.p.) exhibited significant anxiolytic actions in WT mice except in KO mice. In only WT mice, significant antidepressant effects were observed in tail suspension test (TST) and forced swim test (FST). The multichannel electrophysiological technique demonstrated that YL-IPA08 significantly increased the firing rates of pyramidal neurons and decreased those of interneurons. Further studies illustrated that the firing rates of glutamatergic might be antagonized by PK11195 (a classic TSPO antagonist). Our results suggest that YL-IPA08 might regulate the E:I balance in mPFC, mediated by TSPO. In summary, TSPO regulates E:I functional balance in mPFC, play a critical role in antidepressant-anxiolytic effects of YL-IPA08, and provide a potential target site for the development of antidepressant and anxiolytic drugs.

New prediction method of horizontal principal stress of deep tight reservoirs.

As the tight reservoir of Lucaogou Formation in Jimsar sag is characterized as large burial depth and poor physical properties, hydraulic fracturing technology is greatly needed to increase the production. Firstly, based on elastic theory, the borehole displacement formula under stress is derived, then the borehole quasi-ellipse characteristic model is proposed, on this basis, the in-situ stress prediction model based on borehole deformation is constructed. Secondly, the force analysis of diamond bit during drilling is performed, and the continuous prediction formula of elastic modulus based on weight on bit (WOB) and torque parameters is derived. Combined with these two steps, a new model of continuous in-situ stress prediction based on borehole deformation and drilling parameters is established. Compared with the values measured by the acoustic emission method in the laboratory, the deviation between the predicted in-situ stress values and those measured by the acoustic emission method is less than 4%, which meets the accuracy requirements of the fracturing in the oil field. The prediction results show that the minimum horizontal principal stress of Lucaogou Formation is 51.1-62.7 MPa, and the maximum horizontal principal stress is 58.9-69.1 MPa, respectively. The maximum horizontal principal stress direction is NW-SE direction with an angle of about 159°. The results show that the fracture direction in the study area is in accordance with the present in-situ stress direction, which is beneficial to the secondary reconstruction of natural fractures and keeps good opening property of fractures. This study provides a theoretical basis for in-situ stress prediction and compressibility evaluation of tight reservoir.

Experimental investigation of the mechanical properties of mud shale under water-bearing conditions and its applications.

Mud shale, used in drilling engineering, is prone to hydration and expansion, resulting in creep deformation that leads to wellbore shrinkage and pipe sticking incidents. Studying the creep characteristics of mud shale is significant for designing a reasonable well structure and determining the lower limit of drilling fluid density. The influence of moisture content on rock strength and creep mechanical properties were studied using water absorption, uniaxial compression, and creep tests. Test results show that with an increase in the moisture content, the mud shale was damaged and softened; moreover, the elastic modulus decreased with increase in moisture content. Under the same load level, the instantaneous strain increased with increasing moisture content. Under different loading stresses, the creep of the rock had nonlinear characteristics, which could be divided into three different creep stages: attenuation, second, and accelerated creep. A new improved creep model based on the Nishihara model was established to describe the accelerated creep characteristics of mud shale under different moisture contents. The ageing degradation and water-bearing weakening effects were introduced. The Levenberg-Marquardt nonlinear least-squares method was applied to invert the creep parameters. The results show that the simulated creep curves, generated using the new creep model, conform to the experimental ones. The relationship between the drilling fluid density and wellbore shrinkage ratio can be defined using this model; it provides a reference for reasonably determining the drilling fluid density.

Palladium-Catalyzed Asymmetric Sequential Hydroamination of 1,3-Enynes: Enantioselective Syntheses of Chiral Imidazolidinones.

Pd-catalyzed sequential hydroamination of readily available 1,3-enynes is reported. The redox-neutral process provides an efficient route to synthesize a broad scope of imidazolidinones, thiadiazolidines, and imidazolidines. Asymmetric sequential hydroamination generates a series of synthetically valuable, enantioenriched imidazolidinones. Mechanistic studies revealed that the transformation occurred via an intermolecular enyne hydroamination pathway to give an allene intermediate. Subsequent intramolecular hydroamination of the allene intermediate proceeded under the Curtin-Hammett principle to provide enantioenriched imidazolidinone products.

Green preparation of porous hierarchical TiO2(B)/anatase phase junction for effective photocatalytic degradation of antibiotics.

In this study, porous hierarchical bronze/anatase phase junction TiO2 assembled by ultrathin two-dimensional nanosheets was prepared by a novel, green and simple deep eutectic solvent-regulated strategy. Due to its structural features, the TiO2 sample exhibited enhanced photocatalytic activities for multiple kinds of antibiotics, including ofloxacin, ciprofloxacin and chloramphenicol.

Study on discriminant method of rock type for porous carbonate reservoirs based on Bayesian theory.

Rock typing is an extremely critical step in the estimation of carbonate reservoir quality and reserves in the Middle East. In order to recognize the rock types of carbonate reservoirs in the Mishrif Formation better, classify the reservoirs accurately, and establish the permeability model in line with the study area precisely, it is necessary to study the recognition method conforming to the actual situation of the study area. The practice shows that the current recognition methods based on capillary pressure curve, flow unit and NMR logging data can effectively distinguish rock types, but a large number of accurate experimental data are required, which can only be applied in a few cored well, however, cannot be applied in the whole oil field. In this study, based on core, thin section, logging data, the sedimentary characteristics of carbonate reservoir, logging response of four rock types as well as porosity and permeability characteristics of Mishrif Formation in W are comprehensively studied. Based on Bayesian stepwise discriminant theory in multivariate statistics, the Bayesian discrimination model based on conventional logging data is established. The examining results showed that, compared with the description of logging and coring, the accuracy of Bayesian discriminant model and cross confirmation rate have achieved more than 80% for the original sample. Reliability verification showed that the matching degree of the rock type recognized in the non-cored well with the core and mud logging was as high as 90%, which matched the depositional environment of the entire region. The study results confirm the validity and generalizability of the Bayesian method to identify and predict rock types, which can be applied to the entire Middle East region to solve the problem of the lack of core data to accurately evaluate the quality of non-cored wells and accurately predict production, meeting the needs of actual reservoir evaluation and production development in the Middle East.

Male renal functions are associated with serum short- and medium-chain chlorinated paraffins in residents from Jinan, China.

BACKGROUND: Chlorinated paraffins (CPs) are contaminants ubiquitously detected in environmental samples, and reports addressing CPs in human samples are expanding. While CP exposure was suggested to impair kidney function by in vivo/in vitro experiments, epidemiological evidence is lacking. OBJECTIVE: To examine the associations between serum total short-chain CP and medium-chain CP concentrations (∑SCCPs and ∑MCCPs) with human kidney function. METHODS: The study samples were obtained from 387 participants living in Jinan, North China. We quantified ∑SCCPs and ∑MCCPs in serum samples and evaluated the kidney function of included subjects by estimated glomerular filtration rate (eGFR). The associations between serum ∑SCCPs, ∑MCCPs and eGFR were estimated using multivariable linear regression and logistic regression. The possible gender-dependent effects were studied by stratified analysis. RESULTS: After adjusting for age, education, smoking status, drinking status, body mass index (BMI), family history of chronic kidney disease (CKD), fasting serum glucose, systolic blood pressure and diastolic blood pressure, higher concentrations of serum ∑SCCPs and ∑MCCPs were associated with higher male eGFR (ß = 3.13 mL/min/1.73 m2 per one ln-unit increase of serum ∑SCCPs, 95%CI: 1.72, 4.54, p = 0.016; ß = 3.52 mL/min/1.73 m2 per one ln-unit increase of serum ∑MCCPs, 95%CI: 1.89, 5.17, p = 0.011). Associations between serum ∑SCCPs, ∑MCCPs and female eGFR were null. Comparing higher (above the median serum CP levels) vs. lower exposure groups, serum ∑SCCPs and ∑MCCPs were associated with an elevated risk of glomerular hyperfiltration (GH, eGFR ≥ 135 mL/min/1.73 m2), which was associated with glomerular damage and represented as an early stage of chronic kidney disease (OR = 2.98; 95% CI: 1.24, 4.71 for SCCPs; OR = 3.25; 95% CI: 1.20, 5.29 for MCCPs). CONCLUSIONS: Our study suggests that male serum ∑SCCPs and ∑MCCPs are associated with an increased risk of GH, indicating early-stage kidney impairment.

The association of liver function biomarkers with internal exposure of short- and medium-chain chlorinated paraffins in residents from Jinan, China.

Chlorinated paraffins (CPs) are pervasive environmental pollutants which have been reported to be hepatotoxic by laboratory cell and animal studies. However, the related epidemiological reports on their hepatotoxic effects to humans are sparse. In this study, we evaluated the associations between six liver enzymes and serum short-chain CP (SCCP) or medium-chain CP (MCCP) concentrations of 197 residents in Jinan, China. Serum S/MCCPs were detected by quadrupole time-of-flight high-resolution mass spectrometry coupled with atmospheric pressure chemical ionization source (APCI-QTOF-HRMS), and quantified by pattern deconvolution method. The associations between total serum S/MCCP concentrations (ΣS/MCCPs) and continuous liver enzyme levels were assessed by linear regression. Odds ratios (ORs) for the effects of serum ΣS/MCCPs concentrations on liver function biomarkers dichotomized by clinical reference intervals were predicted by logistic regression, either treating ΣS/MCCPs as continuous or categorical dependents. After multivariable adjustment, linear regression results illustrated that 1-ln unit increase in serum ΣSCCPs was negatively associated with male PA levels [-6.08, 95% confidence interval (CI): -11.90, -3.25, p < 0.05], positively associated with male TB levels (1.80, 95% CI: 0.28, 3.31, p < 0.05), and positively associated with female AST levels (1.39, 95% CI: 0.07, 2.70, p < 0.05). One-ln unit increase in serum ΣMCCPs was negatively associated male PA levels (-7.56, 95% CI: -17.15, -4.03, p < 0.05). Logistic regression results suggested that male serum ΣSCCPs were associated with increased prevalence of abnormal PA (OR = 1.47 per 1 ln-unit increase, CI = 1.18, 1.82) and TB (OR = 1.75, 95% CI = 1.12, 2.76) levels, and male serum ΣMCCPs were significantly associated with increased prevalence of abnormal PA (OR = 1.43, 95% CI = 1.03, 1.97) levels. In addition, male participants with concentrations above the median ΣS/MCCPs were associated with increased risk for abnormal PA levels [SCCPs, 2.11-fold (95% CI = 1.15, 3.87); MCCPs, 1.94-fold (95% CI = 1.24, 3.03)]. Male participants with concentrations above the median ΣSCCPs were also associated with increased risk for abnormal TB levels (OR = 1.75, 95% CI = 1.12, 2.76). Conclusively, our results revealed that CP internal exposure was associated with disturbed liver biomarker levels, suggesting the hepatotoxicity of both SCCPs and MCCPs to humans.

The Faster-Onset Antidepressant Effects of Hypidone Hydrochloride (YL-0919) in Monkeys Subjected to Chronic Unpredictable Stress.

Given the limited monkey models of depression available to date, as well as the procedural complexity and time investments that they involve, the ability to test the efficacy and time course of antidepressants in monkey models is greatly restricted. The present study attempted to build a simple and feasible monkey model of depression with chronic unpredictable stress (CUS) and evaluate the antidepressant effect and onset time of fluoxetine hydrochloride (FLX) and the new drug hypidone hydrochloride (YL-0919), a potent and selective 5-HT reuptake inhibitor, 5-HT1A receptor partial agonist and 5-HT6 receptor full agonist. Female cynomolgus monkeys with low social status in their colonies were selected and subjected to CUS for 8 weeks by means of food and water deprivation, space restriction, loud noise, strobe light, and intimidation with fake snakes. Huddling, self-clasping, locomotion and environmental exploration were monitored to evaluate behavioral changes. In addition, the window-opening test was used to evaluate the exploratory interest of the monkeys. The present results revealed that CUS-exposed monkeys displayed significant depression-like behaviors, including significant decreases in exploratory interest, locomotion, and exploration as well as significant increases in huddling and self-clasping behavior and the level of fecal cortisol after 8 weeks of CUS. Treatment with FLX (2.4 mg/kg, i. g.) or YL-0919 (1.2 mg/kg, i. g.) markedly reversed the depression-like behaviors caused by CUS, producing significant antidepressant effects. YL-0919 (once daily for 9 days) had a faster-onset antidepressant effect, compared with FLX (once daily for 17 days). In summary, the present study first established a CUS model using female cynomolgus monkeys with low social status and then successfully evaluated the onset time of 5-HTergic antidepressants. The results suggested that monkeys exposed to CUS displayed significant depression-like behaviors, and both FLX and YL-0919 produced antidepressant effects in this model. Moreover, YL-0919 appeared to act faster than FLX. The present study provides a promising prospect for the evaluation of fast-onset antidepressant drugs based on a CUS monkey model.

Controllable fabrication of TiO2 anatase/rutile phase junctions by a designer solvent for promoted photocatalytic performance.

Herein, novel coin tree-like TiO2 moieties with lots of anatase-rutile phase junctions were constructed by a general, simple, and environmentally friendly strategy. The anatase/rutile ratios can be easily tuned by changing the ratios of the two H-bond donors. Owing to this featured shape, the TiO2 sample displays robust photocatalytic activity and better stability.

Serotonergic transmission is required for the anxiolytic-like behavioral effects of YL-IPA08, a selective ligand targeting TSPO.

Anxiety disorders are the most prevalent group of mental disorders globally, leading to considerable losses in health, functioning and increase of medical costs. Till now, the search for novel pharmacological treatments is driven by the growing medical need to improve on the effectiveness and the side effect profile of existing drugs. In central nervous system, the mitochondrially located translocator protein (18 kDa, TSPO) serves as the rate-limiting step for neurosteroidogenesis and influences GABAergic transmission. Since 5-HT is one of the most comprehensively studied neurotransmitter systems in the anxiety field, in the present study, we want to investigate whether 5-HT system is involved in the anxiolytic-like effects of YL-IPA08, a novel TSPO ligand designed and synthesized at our institute. Our data showed that YL-IPA08 could potentiate the 5-HTP-induced head-twitch response, and the anxiolytic-like effect of YL-IPA08 was abolished by pCPA or 5,7-DHT pretreatment in mice. Furthermore, we found that YL-IPA08 increased the extracellular levels of 5-HT in the rat ventral hippocampus in freely moving rat using the rapid and validated HPLC coupled with microdialysis. In addition, 5-HT level was positively correlated with the level of allopregnanolone. The above results suggest that 5-HT neurotransmission may play a critical role in the anxiolytic-like effects of YL-IPA08.

Short and medium-chain chlorinated paraffins in serum from residents aged from 50 to 84 in Jinan, China: Occurrence, composition and association with hematologic parameters.

Human exposure to chlorinated paraffins (CPs) has been expected and assessed by external pathways considering their pervasive environmental occurrence. However, the deficiency of external exposure assessment in characterizing human burden is unavoidable. In addition, the associations between health outcomes and CP contents in human biospecimen are rarely assessed. In this study, we reported the occurrence and homologue profiles of short-chain chlorinated paraffins (SCCPs) and medium-chain chlorinated paraffins (MCCPs) in serum samples from 145 residents aged from 50 to 84 in Jinan, Shandong Province of China using quadrupole time-of-flight high-resolution mass spectrometry coupled with atmospheric pressure chemical ionization source operated in negative ion mode (APCI-qTOF-HRMS). The associations between serum CP concentrations and hematologic parameters were further analyzed by linear regression. We identified high level of ∑SCCPs (median = 107 ng/g wet weight, ww; 13,800 ng/g lipid, lw), ∑MCCPs (median = 134 ng/g ww; 15,200 ng/g lw) and elevated ∑MCCPs/∑SCCPs (median = 1.12) in serum of the studied population. C13-CPs and C14-CPs were the most abundant SCCP and MCCP groups, respectively. While the predominant chlorine homologues among SCCPs and MCCPs were Cl7-8-CPs. ∑SCCPs, ∑MCCPs, ∑MCCPs/∑SCCPs and the homologue patterns presented no significant variance among age, sex and BMI groups. Further explorations suggested that perturbation of hematologic homeostasis could be induced by CP exposure in a sex-specific way, reflected by significant negative associations of serum SCCP and MCCP concentrations in lipid weight basis with sex-specific hematologic parameters. This study suffered from some limitations and should be interpreted with caution. However, the CP burdens of residents in China and the subsequent health risks must be underscored.

Rapid anti-PTSD-like activity of the TSPO agonist YL-IPA08: Emphasis on brain GABA, neurosteroids and HPA axis function.

There is a serious need for fast-acting drugs to treat post-traumatic stress disorder (PTSD). Our previous studies revealed that YL-IPA08, a novel small-molecule TSPO agonist, exerted significant anti-PTSD effects in various animal models. However, the onset time of YL-IPA08 and its underlying mechanisms remain unclear. In the present study, we first investigated the time course of YL-IPA08 compared to selective serotonin reuptake inhibitors (SSRIs) in the well-known time-dependent sensitization model of PTSD. YL-IPA08 required only 2-4 days of treatment to take effect in behavioural models of PTSD, whereas sertraline required 7-8 days. Furthermore, the mechanism study revealed that YL-IPA08 elicited anti-PTSD-like effects associated with increased GABA levels and allopregnanolone efflux in the hippocampus and prefrontal cortex and increased corticosterone levels in the serum after only 5 days of treatment, whereas sertraline required 9 days. Our results demonstrate that YL-IPA08 can exert fast-onset anti-PTSD-like effects, and its mechanisms may be related to the increased GABA levels, allopregnanolone efflux and the hypothalamic-pituitary-adrenal (HPA) axis function.

CircRNAs and its relationship with gastric cancer.

Circular RNAs (circRNAs), as a type of tissue specific RNA with more stable structure than linear RNAs, was poorly understood on its correlation with gastric cancer (GC). In this review, we outline the synthesis and characteristics of circRNAs and generalize their categories and functions. Through comprehensive analysis of the reported results, we find that circRNAs not only participate in the regulation of gastric cancer (GC) cell biological behaviors, such as proliferation, invasion, migration and epithelial mesenchymal transition (EMT), but also are related to the clinicopathological features of GC such as tumor differentiation, TNM stage and metastasis, etc. According to the present screening and verification results, circRNAs are suggested to be used as biomarkers for the early diagnosis and prognosis prediction of GC, and those circRNAs involved in the genesis and development of GC have the potential as novel targets for the individualized treatment of GC.

Four novel polymorphisms in long non-coding RNA HOTTIP are associated with the risk and prognosis of colorectal cancer.

BACKGROUND: The role of long non-coding RNA (lncRNA) HOXA transcript at the distal tip (HOTTIP) as an oncogene in varieties of human cancer including colorectal cancer (CRC) has been extensively researched. The expression and function of lncRNAs could be affected by single nucleotide polymorphisms (SNPs), which are associated with cancer susceptibility and prognosis. However, no investigation has focused on the association between HOTTIP SNPs and CRC. The aim of the present study was to explore the association of polymorphisms in the lncRNA HOTTIP gene with CRC risk and prognosis. METHODS: A total of 1848 subjects were enrolled in our study, including 884 CRC cases and 964 controls. Genotyping for five HOTTIP tagSNPs (rs3807598, rs17501292, rs2067087, rs17427960, and rs78248039) was performed by applying Kompetitive allele specific PCR (KASP). RESULTS: The results showed three SNPs (rs3807598, rs2067087, and rs17427960) were associated with enhanced CRC risk both in overall and stratified analysis. One polymorphism, rs17501292, could improve the overall survival (OS) of CRC patients in the tumor of ulcerative/invasive-type subgroup. CONCLUSION: These findings suggest HOTTIP SNPs could potentially be predictive biomarkers for CRC risk and prognosis. The present study provides clues for further exploration of novel lncRNA-based genetic biomarkers to predict CRC susceptibility as well as clinical outcome.