Preoperative recovery sleep ameliorates postoperative cognitive dysfunction aggravated by sleep fragmentation in aged mice by enhancing EEG delta-wave activity and LFP theta oscillation in hippocampal CA1.

Sleep fragmentation (SF) is a common sleep problem experienced during the perioperative period by older adults, and is associated with postoperative cognitive dysfunction (POCD). Increasing evidence indicates that delta-wave activity during non-rapid eye movement (NREM) sleep is involved in sleep-dependent memory consolidation and that hippocampal theta oscillations are related to spatial exploratory memory. Recovery sleep (RS), a self-regulated state of sleep homeostasis, enhances delta-wave power and memory performance in sleep-deprived older mice. However, it remains unclear whether RS therapy has a positive effect on cognitive changes following SF in older mouse models. Therefore, this study aimed to explore whether preoperative RS can alleviate cognitive deficits in aged mice with SF. A model of preoperative 24-h SF combined with exploratory laparotomy-induced POCD was established in 18-month-old mice. Aged mice were treated with preoperative 6-h RS following SF and postoperative 6-h RS following surgery, respectively. The changes in hippocampus-dependent cognitive function were investigated using behavioral tests, electroencephalography (EEG), local field potential (LFP), magnetic resonance imaging, and neuromorphology. Mice that underwent 24-h SF combined with surgery exhibited severe spatial memory impairment; impaired cognitive performance could be alleviated by preoperative RS treatment. In addition, preoperative RS increased NREM sleep; enhanced EEG delta-wave activity and LFP theta oscillation in the hippocampal CA1; and improved hippocampal perfusion, microstructural integrity, and neuronal damage. Taken together, these results provide evidence that preoperative RS may ameliorate the severity of POCD aggravated by SF by enhancing delta slow-wave activity and hippocampal theta oscillation, and by ameliorating the reduction in regional cerebral blood flow and white matter microstructure integrity in the hippocampus.

A novel prognostic model of breast cancer based on cuproptosis-related lncRNAs.

OBJECTIVE: Breast cancer (BC) is a deadly form of malignancy responsible for the death of a large number of women every year. Cuproptosis is a newly discovered form of cell death that may have implications for the prognosis of BC. Long non-coding RNAs (lncRNAs) have been shown to be involved in the progression and development of BC. Here within, a novel model capable of predicting the prognosis of patients with BC was established based on cuproptosis-related lncRNAs. METHODS: Data of breast cancer patients was downloaded, including clinical information from The Cancer Genome Atlas (TCGA) database and lncRNAs related to cuproptosis were isolated. In total, nine lncRNAs related to copper death were obtained by Cox regression model based on Least Absolute Shrinkage and Selector Operation (LASSO) algorithm for model construction. The model was verified by overall survival (OS), progression-free survival (PFS) and receiver operating characteristic (ROC) curve. The differences in immune function, tumor mutation burden (TMB) and tumor immune dysfunction and exclusion (TIDE) between patients with different risk scores were analyzed. RESULTS: Based on cuproptosis-related lncRNAs, a prognostic model for predicting BC was constructed. Each patient was assigned a risk score based on our model formula. We found that patients with higher risk scores had significantly lower OS and PFS, increased TMB, and higher sensitivity to immunotherapy. CONCLUSIONS: The model established in this study based on cuproptosis-related lncRNAs may be capable of improving the OS of patients with BC.

The locus coeruleus-dorsal hippocampal CA1 pathway is involved in depression-induced perioperative neurocognitive disorders in adult mice.

BACKGROUND: Patients undergoing surgical anesthesia increasingly suffer from preoperative depression. Clinical studies have shown that depression is a risk factor for perioperative neurocognitive disorders (PNDs) in elder patients. However, the underlying mechanism, especially at the neural circuit level, remains poorly understood. METHODS: Right carotid artery separation under sevoflurane and chronic social defeat stress (CSDS) in adult mice were used to establish surgical anesthesia and chronic depression models. Cognitive function was assessed by the Y maze and novel object recognition tests. A chemogenetic approach was used to modulate the locus coeruleus-dorsal hippocampal CA1 (LC-dCA1) circuit. Hippocampal synaptic alterations were evaluated by Golgi staining and whole-cell patch clamp recording. RESULTS: We found that CSDS induced synaptic impairments in dorsal hippocampal CA1 pyramidal neurons and cognitive deficits in adult mice after surgery under sevoflurane. Chemogenetic activation of the LC-dCA1 pathway significantly alleviated the CSDS-induced synaptic impairments and cognitive dysfunction. On the contrary, inhibition of this pathway could mimic CSDS-induced deficits. Furthermore, we showed that dopamine played an important role in CSDS-induced PNDs in adult mice after surgery/sevoflurane. CONCLUSION: Overall, our results have demonstrated a vital role for the LC-dCA1 pathway in CSDS-induced PNDs in adult mice undergoing surgery with sevoflurane anesthesia.

HPFG: semi-supervised medical image segmentation framework based on hybrid pseudo-label and feature-guiding.

Semi-supervised learning methods have been attracting much attention in medical image segmentation due to the lack of high-quality annotation. To cope with the noise problem of pseudo-label in semi-supervised medical image segmentation and the limitations of contrastive learning applications, we propose a semi-supervised medical image segmentation framework, HPFG, based on hybrid pseudo-label and feature-guiding, which consists of a hybrid pseudo-label strategy and two different feature-guiding modules. The hybrid pseudo-label strategy uses the CutMix operation and an auxiliary network to enable the labeled images to guide the unlabeled images to generate high-quality pseudo-label and reduce the impact of pseudo-label noise. In addition, a feature-guiding encoder module based on feature-level contrastive learning is designed to guide the encoder to mine useful local and global image features, thus effectively enhancing the feature extraction capability of the model. At the same time, a feature-guiding decoder module based on adaptive class-level contrastive learning is designed to guide the decoder in better extracting class information, achieving intra-class affinity and inter-class separation, and effectively alleviating the class imbalance problem in medical datasets. Extensive experimental results show that the segmentation performance of the HPFG framework proposed in this paper outperforms existing semi-supervised medical image segmentation methods on three public datasets: ACDC, LIDC, and ISIC. Code is available at https://github.com/fakerlove1/HPFG .

Coupling effect of reaction conditions on the catalytic activity of Cu-Mn composite oxide catalysts for toluene.

Volatile organic compounds (VOCs) are one of the major components of air pollution. Catalytic combustion is a promising technology for the treatment of VOCs and at its center is the preparation of efficient and cheap catalysts. In this study, by loading copper (Cu) and manganese (Mn) on Santa Barbara Amorphous-15 (SBA-15) molecular sieve, the Cux-Mny/SBA-15 (x = 1, 2; y = 1, 2) composite metal oxide catalyst was prepared using the equal volume impregnation method. Their performance in the toluene catalytic combustion reaction was investigated by adjusting the molar ratio (x : y), and the loading of Cu and Mn. The results of the Brunner-Emmett-Teller (BET), X-ray diffraction (XRD), scanning electron microscopy (SEM), and X-ray photoelectron spectroscopy (XPS) analyses show that the CuMnO spinel phase can be detected in the Cu-Mn composite metal oxide catalyst doped with a low concentration of Cu. The overall rod-like structure of the fibrous network structure provides a large specific surface area, and the particle crystallinity is low and the dispersion is good. Due to the synergistic effect of Cu and Mn, the greater the amount of Mn3+ and adsorbed oxygen species (Oads) that are available, and the higher the turnover frequency (TOF) value, the better and more superior catalytic performance and excellent stability is obtained, when compared with the single-component oxides used in toluene catalytic combustion. After a continuous catalytic reaction for 12 h, the toluene conversion rate remained above 95%. The coupling effect of the catalytic reaction temperature and concentration of oxygen on the catalytic combustion of toluene was also studied. At a low reaction temperature (<250 °C), the increase of the concentration of oxygen played a superior role in promoting the conversion of toluene. The kinetic analysis of the toluene catalytic combustion process showed that the catalytic combustion of toluene by Cu-Mn/SBA-15 followed both the Mars-Van Krevelen (MVK) and Langmuir-Hinshelwood (L-H) reaction mechanisms. With the increase of the Oads amount caused by the decrease of the Cu ratio, the proportion of the L-H reaction mechanism increases.

Analysis of Magnetic Resonance Imaging Perfusion Parameters for the Identification of Spinal Metastatic Tumors with Rich Blood Supply.

PURPOSE: To determine the reliability of dynamic magnetic resonance imaging (MRI) perfusion parameters for the evaluation of blood supply to spinal metastatic tumors. METHODS: A total of 36 patients with spinal metastasis who underwent dynamic contrast-enhanced magnetic resonance spinal perfusion imaging at Tianjin Hospital from December 2018 to December 2020 were reviewed. Subsequently, the patients underwent corresponding preoperative examination using digital subtraction angiography of the spine at the hospital and were divided into 2 groups accordingly. Differences in dynamic MRI perfusion parameters between the 2 groups were analyzed. RESULTS: There were statistically significant differences between the 2 groups in the quantitative dynamic contrast-enhanced MRI perfusion parameters vascular permeability and plasma volume, as well as semi-quantitative peak enhancement and blood flow ratio parameters. CONCLUSIONS: Dynamic MRI perfusion may distinguish spinal metastatic lesions with rich blood supply from those with poor blood supply and may help clinicians identify patients that can benefit from invasive spinal angiography and preoperative embolization. This technique may also provide guidance on decision taking for surgery basing on dynamic MRI perfusion parameters.

ALKBH5 ALLEVIATES HYPOXIA POSTCONDITIONING INJURY IN d -GALACTOSE-INDUCED SENESCENT CARDIOMYOCYTES BY REGULATING STAT3.

ABSTRACT: Ischemic postconditioning (I/Post) reduces I/R injury by activating endogenous cardioprotection mechanisms, such as the JAK/signal transducer and activator of transcription 3 (STAT3) and PI3K/Akt pathways, which offer a traditional approach to myocardial protection. According to a previous study, cardioprotection by I/Post may be lost in aged mice, and in our previous research, hypoxic postconditioning (H/Post) lacked a protective effect in senescent cardiomyocytes, which was associated with low expression of long noncoding RNA H19. The N6-methyladenosine (m 6 A) modification is a dynamic and reversible process that has been confirmed to play a role in cardiovascular diseases. However, the mechanisms of m 6 A modification in myocardial I/Post remain to be explored. Neonatal cardiomyocytes were isolated from 2-day-old Sprague-Dawley rats, and senescence was induced by d -galactose, followed by stimulation of hypoxia-reoxygenation and H/Post. Hypoxic injury was evaluated by cell viability and the Bcl-2/Bax protein ratio. Total m 6 A levels were measured using a colorimetric m 6 A RNA Methylation Quantification Kit, and the m 6 A modified and differentially expressed mRNA was determined by MeRIP (methylated RNA immunoprecipitation). We found that H/Post increased m 6 A methylation and decreased RNA mA demethylase alkB homolog 5 (ALKBH5) expression in aged cardiomyocytes. Furthermore, ALKBH5 knockdown exacerbated injury following H/Post in senescent cardiomyocytes. In addition, ALKBH5 regulated STAT3 expression by mediating its m 6 A modification and long noncoding RNA H19/miR-124-3p. ALKBH5 also alleviated the H/Post injury induced by the low expression of STAT3 in senescent cardiomyocytes.

Effect of Folic Acid Treatment for Patients with Traumatic Brain Injury (TBI)-Related Hospital Acquired Pneumonia (HAP): A Retrospective Cohort Study.

Hospital Acquired Pneumonia (HAP) is one of the most common complications and late causes of death in TBI patients. Targeted prevention and treatment of HAP are of great significance for improving the prognosis of TBI patients. In the previous clinical observation, we found that folic acid treatment for TBI patients has a good effect on preventing and treating HAP. We conducted this retrospective cohort study to demonstrate what we observed by selecting 293 TBI patients from two medical centers and analyzing their hospitalization data. The result showed that the incidence of HAP was significantly lower in TBI patients who received folic acid treatment (44.1% vs. 63.0%, p = 0.012). Multivariate logistic regression analysis showed that folic acid treatment was an independent protective factor for the occurrence of HAP in TBI patients (OR = 0.418, p = 0.031), especially in high-risk groups of HAP, such as the old (OR: 1.356 vs. 2.889), ICU (OR: 1.775 vs. 5.996) and severe TBI (OR: 0.975 vs. 5.424) patients. At the same time, cohort studies of HAP patients showed that folic acid also had a good effect on delaying the progression of HAP, such as reducing the chance of tracheotomy (26.1% vs. 50.8%, p = 0.041), and reduced the length of hospital stay (15 d vs. 19 d, p = 0.029) and ICU stay (5 d vs. 8 d, p = 0.046). Therefore, we believe that folic acid treatment in TBI patients has the potential for preventing and treating HAP, and it is worthy of further clinical research.

Age-dependent Electroencephalogram Characteristics During Different Levels of Anesthetic Depth.

Objective The monitoring of anesthetic depth based on electroencephalogram derivation is not currently adjusted for age. Here we analyze the influence of age factors on electroencephalogram characteristics. Methods Frontal electroencephalogram recordings were obtained from 80 adults during routine clinical anesthesia. The characteristics of electroencephalogram with age and anesthesia were observed during four kinds of anesthesia. Results The slow wave power, δ power, Bispectral Index (BIS) and approximate entropy can be used to distinguish different states of anesthesia (P < 0.05). In the deep and very deep anesthesia states, δ power decreased with age (P < 0.0001). In the very deep anesthesia state, θ power decreased with age (P < 0.05). In the deep and very deep anesthesia states, α power decreased with age (P = 0.0002). In the light and deep anesthesia states, ß power decreased with age (P = 0.003). In the deep anesthesia state, γ power decreased with age (P = 0.002). In the very deep anesthesia state, permutation entropy increased significantly with age (P = 0.0001). In the very deep anesthesia state, BIS value increased with age (P = 0.006). The slow wave power, approximate entropy, and sample entropy did not show age-dependent changes. Conclusions The influence of age should be considered when using BIS and δ power to monitor the depth of anesthesia, while the influence of age should not be considered when using slow wave power and approximate entropy to monitor the depth of anesthesia.

DPBET: A dual-path lung nodules segmentation model based on boundary enhancement and hybrid transformer.

Accurate segmentation of lung nodules is an important basis for the subsequent differentiation of benign and malignant pathological types, which is conducive to early detection of lung cancer. Due to the local feature extraction characteristics of convolution and the limited receptive field of continuous down-sampling, the existing deep convolutional neural networks (CNN) for lung nodules segmentation cause the loss of information of lesion boundaries and locations. To address this issue, a dual-path lung nodules segmentation model based on boundary enhancement and hybrid transformer (DPBET) is proposed in this paper. The model consists of a global path, an edge path, and a feature aggregation module. In the global path, a de-redundant transformer module with explicit guidance is proposed, called Cascade-Axial-Prune Transformer (CAP-Trans). It is combined with CNN to form a hybrid architecture to generate a global representation of the target lesion. In the edge path, an edge detection operator is introduced to construct a lung nodule edge enhancement dataset, which improves the dataset utilization while providing more prior knowledge of the target lesion boundar. In addition, the Down-Attention Sample (DASample) as a basic encoding block is designed to effectively perceive local features of different ranges and scales in the down-sampling process of lung nodule feature extraction. Finally, a feature aggregation module is designed to fuse the outputs of the two paths to get the final segmentation result. Our DPBET can delineate the boundaries of various types of pulmonary nodules, with an average DSC of 89.86% and an average Sensitivity of 90.50% on the public dataset LIDC-IDRI. Compared with the state-of-the-art approaches, a substantial improvement has been achieved. The experimental results demonstrate that DPBET can use edge enhancement to promote the global-edge consistency relationship, and the network architecture is effective in lung nodule segmentation.

Co-combustion of high alkali coal with municipal sludge: Thermal behaviour, kinetic analysis, and micro characteristic.

Blending sludge rich in protein and aliphatic hydrocarbons into the high alkali coal (HAC) has been demonstrated to reduce the ash melting temperature of the HAC/sludge mixture, thereby increasing the effectiveness and efficiency of liquid slagging. However, whether the incorporation of sludge can affect the combustion stability of the original coal-fired boiler is still debatable. As the combustion stability of the fuel can directly affect the operational safety of the boiler, it is of great practical value for exploring the effect of sludge incorporation on the combustion performance of HAC. In this work, the thermal behaviour and microscopic properties of individual HAC, municipal sludge (MS) and HAC/MS mixtures were tested using a Thermogravimetric analyser (TGA) and a Fourier transform infrared (FTIR) spectrometer, respectively. The exothermic, thermodynamic and functional group evolution patterns during the combustion of these samples were also evaluated. Ignition temperatures (Ti) of the HAC/MS mixtures were relatively lower than that of individual HAC, and decreased with the increase in sludge mass ratio (SMR). The synergistic effect of the co-combustion of HAC and MS resulted in a slightly higher total heat release during the combustion of MS10HAC90 (i.e., the mass percentage of MS and HAC is 1:9) than HAC alone, however, the total heat release of the blend decreased progressively with increasing SMR. The experimental values of the average Eα for all four mixtures were lower than the theoretical values, indicating that the addition of MS lowered the reaction energy barriers of the mixtures. Consumption rates of the principal groups in samples during the oxidation and combustion all tended to increase progressively with increasing SMR. There are three major synergistic effects existing during co-combustion of HAC and MS: (1) the reaction of free radicals with benzene molecules; (2) the interaction of free radicals; and (3) the catalytic effect of alkali and alkaline earth metals. These findings can provide theoretical guidance for the determination of key parameters (mixing ratio) for the blending of HAC and MS, and can fill the research gap in terms of microscopic reactivity and synergistic effects during the co-combustion of HAC and MS.

[Effects of Different Aeration Treatments on Bacterial Diversity, Metabolic Activity, and Function in Constructed Wetlands].

Bacteria play a key role in the removal of pollutants and nutrients in constructed wetlands. DNA and RNA high-throughput sequencing was used to investigate the diversity, metabolic activity, and function of bacteria in aquaculture wastewater and in constructed wetlands treated by different aeration levels. The results revealed that:① a total of 4042 operational taxonomic units (OTUs) were detected in aquaculture wastewater and constructed wetland treatment groups. α-Proteobacteria, γ-Proteobacteria, and Bacteroidia were the most diverse groups, and the constructed wetlands aeration treatment increased the bacterial diversity to a variable extent; ② α-Proteobacteria, γ-Proteobacteria, Bacteroidia, and Actinobacteria were the dominant groups both in the DNA and RNA sequencing results, and the metabolic activities of these four groups were significantly affected by the concentration of total nitrogen (TN) and nitrate nitrogen (NO3--N) in our study. ③ According to the FAPROTAX database, 56 bacterial functional groups were detected in our study, mainly including:chemoheterotrophy, aerobic chemoheterotrophy, fermentation, intracellular parasites, dark hydrogen oxidation, phototrophy, photoheterotrophy, and nitrate reduction. Functions related to the nitrogen cycle were observed in the results of function annotation, suggesting the important role of bacterial communities in the removal of nitrogen nutrients in constructed wetlands. These results will improve the understanding of bacterial community structures and functions during nutrient removal in aerated constructed wetlands.

Development and validation of a novel risk assessment model to estimate the probability of pulmonary embolism in postoperative patients.

Pulmonary embolism (PE) is a leading cause of mortality in postoperative patients. Numerous PE prevention clinical practice guidelines are available but not consistently implemented. This study aimed to develop and validate a novel risk assessment model to assess the risk of PE in postoperative patients. Patients who underwent Grade IV surgery between September 2012 and January 2020 (n = 26,536) at the Affiliated Dongyang Hospital of Wenzhou Medical University were enrolled in our study. PE was confirmed by an identified filling defect in the pulmonary artery system in CT pulmonary angiography. The PE incidence was evaluated before discharge. All preoperative data containing clinical and laboratory variables were extracted for each participant. A novel risk assessment model (RAM) for PE was developed with multivariate regression analysis. The discrimination ability of the RAM was evaluated by the area under the receiver operating characteristic curve, and model calibration was assessed by the Hosmer-Lemeshow statistic. We included 53 clinical and laboratory variables in this study. Among them, 296 postoperative patients developed PE before discharge, and the incidence rate was 1.04%. The distribution of variables between the training group and the validation group was balanced. After using multivariate stepwise regression, only variable age (OR 1.070 [1.054-1.087], P < 0.001), drinking (OR 0.477 [0.304-0.749], P = 0.001), malignant tumor (OR 2.552 [1.745-3.731], P < 0.001), anticoagulant (OR 3.719 [2.281-6.062], P < 0.001), lymphocyte percentage (OR 2.773 [2.342-3.285], P < 0.001), neutrophil percentage (OR 10.703 [8.337-13.739], P < 0.001), red blood cell (OR 1.872 [1.384-2.532], P < 0.001), total bilirubin (OR 1.038 [1.012-1.064], P < 0.001), direct bilirubin (OR 0.850 [0.779-0.928], P < 0.001), prothrombin time (OR 0.768 [0.636-0.926], P < 0.001) and fibrinogen (OR 0.772 [0.651-0.915], P < 0.001) were selected and significantly associated with PE. The final model included four variables: neutrophil percentage, age, malignant tumor and lymphocyte percentage. The AUC of the model was 0.949 (95% CI 0.932-0.966). The risk prediction model still showed good calibration, with reasonable agreement between the observed and predicted PE outcomes in the validation set (AUC 0.958). The information on sensitivity, specificity and predictive values according to cutoff points of the score in the training set suggested a threshold of 0.012 as the optimal cutoff value to define high-risk individuals. We developed a new approach to select hazard factors for PE in postoperative patients. This tool provided a consistent, accurate, and effective method for risk assessment. This finding may help decision-makers weigh the risk of PE and appropriately select PE prevention strategies.

miR-346-3p promotes osteoclastogenesis via inhibiting TRAF3 gene.

MicroRNAs (miRNAs) modulate gene expression and regulate many physiological and pathological conditions. However, their modulation and effect in osteoclastogenesis remain unknown. In this study, we investigated the role of miR-346-3p in regulating the osteoclast differentiation from RAW264.7 cells. We used the miRNA microarray assay, miR-346-3p mimic transfection, tartrate resistant acid phosphatase (TRAP) staining, bone resorption assay, qRT-PCR, and western blot. Our results showed that the expression of miR-346-3p was significantly upregulated during osteoclast differentiation. Further, by transfecting cells with miR-346-3p mimic, we observed an increased number of TRAP-positive multinucleated cells, increased pit area caused by bone resorption, and enhanced expression of osteoclast-specific genes and proteins. Conversely, miR-346-3p inhibition attenuated the osteoclast differentiation and function. Software-mediated prediction and validation using luciferase reporter assay showed that TRAF3, a negative regulator of osteoclast differentiation, was inhibited by miR-346-3p overexpression. Our results showed that miR-346-3p directly targeted TRAF3 mRNA via binding to its 3'-UTR and inhibited the expression of TRAF3 protein. Taken together, our results revealed that miR-346-3p promotes the regulation of osteoclastogenesis by suppressing the TRAF3 gene. In conclusion, miR-346-3p could be a novel therapeutic target for bone loss-related pathogenesis.

Coupled effects of urbanization level and dam on microplastics in surface waters in a coastal watershed of Southeast China.

Investigating the heterogeneous distribution pattern of plastic pollution in river systems is useful for better understanding the transport pathways of plastics from land to sea. Therefore, we analyzed samples from 17 sites to evaluate the distribution pattern of microplastics (MPs) in Minjiang River Watershed. The MP concentrations ranged between 0.12 and 2.72 n/L, with higher microplastic concentrations located in urban areas with greater population and gross domestic product (GDP) densities. MP concentrations were positively correlated with population density, GDP density, and built-up land use percentage, suggesting that increasing urbanization level will lead to greater plastic pollution. MP concentrations spanning the upstream and downstream sides of the SK Hydropower Dam showed decreasing trends, suggesting that MP settling would occur behind the dam and Shuikou Reservoir might serve as a sink for MPs. This study provides useful insight for revealing the effects of urbanization and dams on spatial distribution of riverine MPs.

Characterization and role of PGK from Litopenaeus vannamei in WSSV infection.

Phosphoglycerate kinase (EC 2.7.2.3, PGK) catalyses the reversible transfer of a phosphate group from 1,3-diphosphoglyceric acid and ADP to produce 3-phosphoglyceric acid and ATP, which represents the initial production of ATP during glycolysis; therefore, PGK is a key enzyme in the energy metabolism. To study the role of PGK in the resistance to WSSV infection in shrimp, the full-length cDNA of the PGK gene (LvPGK) from Litopenaeus vannamei was obtained by using homology cloning and RACE amplification. The tissue distribution of LvPGK and its expression changes in the main immune tissues after WSSV stimulation were obtained by quantitative real-time PCR. Furthermore, RNA interference (RNAi) was used to study the role of LvPGK in shrimp defending against WSSV infection. The results showed that the full-length cDNA sequence of LvPGK was 1855 bp, contained a 1248 bp open reading frame (ORF) encoding 415 amino acids, and included a conserved PGK domain. LvPGK presented ubiquitous expression in most examined tissues, with the most predominant expression in the muscle and the weakest expression in the intestine. LvPGK transcripts could be induced in the hemocytes and hepatopancreas by injection with WSSV. Both the replication of WSSV and the shrimp cumulative mortality decreased significantly after LvPGK knockdown (P < 0.01). After challenging LvPGK RNAi shrimp with WSSV, the concentration of glucose in the hepatopancreas and muscle tissue did not show significant change; however, the content of pyruvate and lactate decreased significantly (P < 0.05). Moreover, significant decreases in the expression levels of crustin, ALF1, ALF2 and ALF3 were also detected. The results suggested that LvPGK might be involved in WSSV replication by increasing host aerobic and anaerobic metabolism.

Glycogen phosphorylase of shrimp (Litopenaeus vannamei): Structure, expression and anti-WSSV function.

Glycogen phosphorylase (GP, EC 2.4.1.1) catalyze the rate-limiting step in glycogenolysis in animals, forming glucose-1-phosphate from the terminal alpha-1,4-glycosidic bond. Therefore, GP plays a crucial role in carbohydrate metabolism. In the present study, the full-length cDNA sequence of GP (LvGP) was cloned from shrimp, Litopenaeus vannamei. The obtained 3242-bp LvGP cDNA sequence included a 5'-terminal untranslated region (UTR) of 48 bp, an open reading frame (ORF) of 2559 bp encoding a polypeptide of 852 amino acids (aa) and a 3'-UTR of 635 bp. The predicted LvGP protein sequence contained a typical phosphorylase domain (113-829 aa) and shared 72%-97% identities with GP from other species. Phylogenetic analysis revealed that LvGP showed the closest relationship with GP from Marsupenaeus japonicus. Tissue expression profiles showed that LvGP existed in most examined tissues, with the most predominant expression in the brain, followed by the muscles and stomach. LvGP transcripts in hepatopancreas and hemocytes were up regulated after the WSSV challenge. Furthermore, the role of LvGP in shrimp defending against WSSV infection was investigated by RNA interference (RNAi). Our findings showed that WSSV proliferation and shrimp accumulative mortality increased significantly after LvGP RNAi (P < 0.01). The glycogen, glucose, and pyruvate content decreased in GP RNAi shrimp after WSSV injection, however, the lactate and ATP concentration enhanced. Moreover, lectin and anti-lipopolysaccharide factor2 (ALF2) were induced in LvGP silencing shrimp after WSSV infection, whereas the expression levels of crustin, ALF1 and ALF3 decreased. Our results suggested that the LvGP might play a crucial role in shrimp defending against WSSV infection by regulating metabolism and affecting the anti-infectious gene expression.

Nematicidal amide alkaloids from the seeds of Clausena lansium.

Five new amide alkaloids (1-3, 5-6) were isolated from the seeds of Clausena lansium together with one new natural product (4) and four known analogues (7-10). The structures of the new amide alkaloids were elucidated based on a comprehensive spectroscopic data analysis including 1D and 2D NMR as well as HRESIMS, and by comparison with the literature. The bioactivity results showed that compound 8 expressed potent nematicidal activity against Panagrellus redivevus, with IC50 value of 0.12 mM, while compounds 3 and 5 presented moderate nematicidal activity with IC50 values of 2.75 and 3.93 mM, respectively (abamectin as the positive control with IC50 value of 1.05 mM).

Three new flavanoids from artificially induced dragon's blood of Dracaena cambodiana.

Three new flavanoids, (2R)-7,4'-dihydroxy-8-methylflavan (1), (2R)-7,4'-dihydroxy-6-methylflavan (2), and (3R)-7,3',4'-trihydroxyhomoisoflavan (3), together with seven known compounds (4-10), were isolated from artificially induced dragon's blood of Dracaena cambodiana, and their structures were determined based on HR-ESI-MS and extensive spectroscopic techniques (UV, IR, 1D-, and 2D-NMR). Compound 2 exhibited weak cytotoxicity against BEL-7402 cells line with the IC50 value of 39.2 µM. In addition, compound 3 showed significant acetylcholinesterase (AChE) inhibitory activity.