Activation of NLRP3 inflammasome in a rat model of cerebral small vessel disease.

Cerebral small vessel disease (CSVD) is increasingly being recognized as a leading contributor to cognitive impairment in the elderly. However, there is a lack of effective preventative or therapeutic options for CSVD. In this exploratory study, we investigated the interplay between neuroinflammation and CSVD pathogenesis as well as the cognitive performance, focusing on NLRP3 signaling as a new therapeutic target. Spontaneously hypertensive stroke-prone (SHRSP) rats served as a CSVD model. We found that SHRSP rats showed decline in learning and memory abilities using morris water maze test. Activated NLRP3 signaling and an increased expression of the downstream pro-inflammatory factors, including IL (interleukin)-6 and tumor necrosis factor α were determined. We also observed a remarkable increase in the production of pyroptosis executive protein gasdermin D, and elevated astrocytic and microglial activation. In addition, we identify several neuropathological hallmarks of CSVD, including blood-brain barrier breakdown, white matter damage, and endothelial dysfunction. These results were in correlation with the activation of NLRP3 inflammasome. Thus, our findings reveal that the NLRP3-mediated inflammatory pathway could play a central role in the pathogenesis of CSVD, presenting a novel target for potential CSVD treatment.

Multi-stage nuclear transcriptomic insights of morphogenesis and biparental role changes in Lentinula edodes.

Based on six offspring with different mitochondrial (M) and parental nuclear (N) genotypes, the multi-stage morphological characteristics and nuclear transcriptomes of Lentinula edodes were compared to investigate morphogenesis mechanisms during cultivation, the key reason for cultivar resistance to genotype changes, and regulation related to biparental role changes. Six offspring had specific transcriptomic data and morphological characteristics that were mainly regulated by the two parental nuclei, followed by the cytoplasm, at different growth stages. Importing a wild N genotype easily leads to failure or instability of fruiting; however, importing wild M genotypes may improve cultivars. Major facilitator superfamily (MFS) transporter genes encoding specific metabolites in spawns may play crucial roles in fruiting body formation. Pellets from submerged cultivation and spawns from sawdust substrate cultivation showed different carbon metabolic pathways, especially in secondary metabolism, degradation of lignin, cellulose and hemicellulose, and plasma membrane transport (mainly MFS). When the stage of small young pileus (SYP) was formed on the surface of the bag, the spawns inside were mainly involved in nutrient accumulation. Just broken pileus (JBP) showed a different expression of plasma membrane transporter genes related to intracellular material transport compared to SYP and showed different ribosomal proteins and cytochrome P450 functioning in protein biosynthesis and metabolism than near spreading pileus (NSP). Biparental roles mainly regulate offspring metabolism, growth, and morphogenesis by differentially expressing specific genes during different vegetative growth stages. Additionally, some genes encoding glycine-rich RNA-binding proteins, F-box, and folliculin-interacting protein repeat-containing proteins may be related to multi-stage morphogenesis. KEY POINTS: • Replacement of nuclear genotype is not suitable for cultivar breeding of L. edodes. • Some genes show a biparental role-divergent expression at mycelial growth stage. • Transcriptomic changes of some sawdust substrate cultivation stages have been elucidated.

Serum Neurofilament Light Chain in Wilson's Disease: A Promising Indicator but Unparallel to Real-Time Treatment Response.

BACKGROUND: Wilson's disease (WD) currently lacks a promising indicator that could reflect neurological impairment and monitor treatment outcome. We aimed to investigate whether serum neurofilament light chain (sNfL) functions as a candidate for disease assessment and treatment monitoring of WD. METHODS: We assessed preclinical and manifested WD patients' sNfL levels compared to controls and analyzed the differences between patients with various clinical symptoms. We then explored the correlation between clinical scales and sNfL levels. And repeated measurements were performed in 34 patients before and after treatment. RESULTS: WD patients with neurological involvement had significantly higher sNfL levels than both hepatic patients and controls. Positive correlations were found between Unified Wilson's Disease Rating Scale scores and sNfL and between semiquantitative magnetic resonance imaging scales and sNfL levels in WD patients. However, in the treatment follow-up analysis, the trend of sNfL before and after treatment disaccorded with clinical response. CONCLUSION: These findings suggest that sNfL levels can be an ideal indicator for the severity of neurological involvement but fail to evaluate change in disease condition after treatment. © 2022 International Parkinson and Movement Disorder Society.

A Parallel Classification Model for Marine Mammal Sounds Based on Multi-Dimensional Feature Extraction and Data Augmentation.

Due to the poor visibility of the deep-sea environment, acoustic signals are often collected and analyzed to explore the behavior of marine species. With the progress of underwater signal-acquisition technology, the amount of acoustic data obtained from the ocean has exceeded the limit that human can process manually, so designing efficient marine-mammal classification algorithms has become a research hotspot. In this paper, we design a classification model based on a multi-channel parallel structure, which can process multi-dimensional acoustic features extracted from audio samples, and fuse the prediction results of different channels through a trainable full connection layer. It uses transfer learning to obtain faster convergence speed, and introduces data augmentation to improve the classification accuracy. The k-fold cross-validation method was used to segment the data set to comprehensively evaluate the prediction accuracy and robustness of the model. The evaluation results showed that the model can achieve a mean accuracy of 95.21% while maintaining a standard deviation of 0.65%. There was excellent consistency in performance over multiple tests.

Improved Self-Organizing Map-Based Unsupervised Learning Algorithm for Sitting Posture Recognition System.

As the intensity of work increases, many of us sit for long hours while working in the office. It is not easy to sit properly at work all the time and sitting for a long time with wrong postures may cause a series of health problems as time goes by. In addition, monitoring the sitting posture of patients with spinal disease would be beneficial for their recovery. Accordingly, this paper designs and implements a sitting posture recognition system from a flexible array pressure sensor, which is used to acquire pressure distribution map of sitting hips in a real-time manner. Moreover, an improved self-organizing map-based classification algorithm for six kinds of sitting posture recognition is proposed to identify whether the current sitting posture is appropriate. The extensive experimental results verify that the performance of ISOM-based sitting posture recognition algorithm (ISOM-SPR) in short outperforms that of four kinds of traditional algorithms including decision tree-based (DT), K-means-based (KM), back propagation neural network-based (BP), self-organizing map-based (SOM) sitting posture recognition algorithms. Finally, it is proven that the proposed system based on ISOM-SPR algorithm has good robustness and high accuracy.

Thermodynamic Evaluation and Sensitivity Analysis of a Novel Compressed Air Energy Storage System Incorporated with a Coal-Fired Power Plant.

A novel compressed air energy storage (CAES) system has been developed, which is innovatively integrated with a coal-fired power plant based on its feedwater heating system. In the hybrid design, the compression heat of the CAES system is transferred to the feedwater of the coal power plant, and the compressed air before the expanders is heated by the feedwater taken from the coal power plant. Furthermore, the exhaust air of the expanders is employed to warm partial feedwater of the coal power plant. Via the suggested integration, the thermal energy storage equipment for a regular CAES system can be eliminated and the performance of the CAES system can be improved. Based on a 350 MW supercritical coal power plant, the proposed concept was thermodynamically evaluated, and the results indicate that the round-trip efficiency and exergy efficiency of the new CAES system can reach 64.08% and 70.01%, respectively. Besides, a sensitivity analysis was conducted to examine the effects of ambient temperature, air storage pressure, expander inlet temperature, and coal power load on the performance of the CAES system. The above work proves that the novel design is efficient under various conditions, providing important insights into the development of CAES technology.

Whole exome sequencing reveals two novel compound heterozygous mutations in TWNK as a cause of the hepatocerebral form of mitochondrial DNA depletion syndrome: a case report.

BACKGROUND: Although Mitochondrial DNA depletion syndrome (MDS) can be classified into three forms: myopathic, encephalomyopathic and hepatocerebral form, it is difficult to identify its form due to its clinical heterogeneity. Therefore, it is very important to conduct molecular genetic analysis on suspected patients. This study presented a male 38 weeks and 5 days infant with liver cytolysis and leukodystrophy. CASE PRESENTATION: A male infant proband was admitted to the department of NICU for feeding intolerance, irregular rhythm of respiration, hypoglycemia, lactic acidosis, liver cytolysis and neurological abnormalities. He was onset of mild jaundice with leukodystrophy and high lactate and phenylderivatives for urine organic acids on the 7th day. Whole exome sequencing (WES) and Sanger sequencing were performed to screen and confirm the suspicious pathogenic mutations. The results revealed this proband carried two compound heterozygous mutations in TWNK: c.1186 C > T / p.Pro396Ser and c.1844 G > C / p.Gly615Ala inherited by an autosomal recessive form from his parents, of which protein conservative analysis and structural modeling supported the pathogenicity of the two mutations. Unfortunately, the conditions described above were not improved until he was discharged from the hospital on the 23rd day and died at 4 months of age. CONCLUSIONS: In this study, we investigated a Chinese family with the hepatocerebral form of MDS and conducted WES and Sanger sequencing to explore the causative mutations for this proband born from non-consanguineous and healthy parents. We identified two novel TWNK c.1186 C > T/ c.1844 G > C compound heterozygous mutations which were probably the disease-causing mutations of hepatocerebral form of MDS and described the clinical manifestations of the proband, which expanded the phenotypic spectrum of MDS caused by variants in TWNK. This study also emphasized WES technology can provide the genetic diagnosis of Mendelian genetic disease.

Metabonomics analysis of kidneys in rats administered with chronic low-dose cadmium by ultra-performance liquid chromatography-mass spectrometry.

This study aimed to investigate the nephrotoxicity in rats administered with chronic low-dose cadmium (Cd) by ultra-performance liquid chromatography-mass spectrometry. A total of 40 male Sprague-Dawley rats were randomly assigned to four groups, namely: control; low-dose (0.13 mg/kg·body weight [bw]); middle-dose (0.80 mg/kg·bw); and high-dose (4.89 mg/kg·bw). The rats received CdCl2 daily via drinking water for 24 weeks. Rat kidneys were collected for metabonomics analysis. Principal components analysis and partial least-squares discriminant analysis were used to investigate the metabonomics profile changes in the kidney samples and to screen the potential biomarkers. Ten metabolites were identified in the positive and negative ion modes. Compared with the control group, the intensities of tetranor 12-HETE, uric acid, hypoxanthine, phenylacetylglycine, guanidinosuccinic acid and xanthosine significantly increased (P < 0.01), and those of imidazolelactic acid, lactose 6-phosphate, l-urobilinogen and arachidonic acid significantly decreased (P < 0.01) in the high-dose group. Results showed that exposure to Cd in rats induced oxidative stress to the kidneys and disrupted amino acid metabolism, fatty acid metabolism and energy metabolism.

Construction of a genetic linkage map of Lentinula edodes based on SSR, SRAP and TRAP markers.

Genetic mapping is a basic tool for eukaryotic genomic research. It allows the localization of genes or quantitative trait loci (QTLs) and map-based cloning. In this study, we constructed a linkage map based on DNA samples from a commercial strain L808, including two parental monokaryons and 93 single spore isolates considered with segregating to 1:1:1:1 at four mating types (A1B1, A1B2, A2B1 and A2B2). Using Simple Sequence Repeats (SSR), Sequence Related Amplified Polymorphism (SRAP), Target Region Amplified Polymorphism (TRAP) molecular markers, 182 molecular markers and two mating factors were located on 11 linkage groups (LGs). The total length of the map was 948.083 centimorgan (cM), with an average marker interval distance of 4.817 cM. Only two gaps spanning more than 20 cM was observed. The probability of 20 cM, 10 cM, 5 cM genetic distance cover one marker was 99.68%, 94.36%, 76.43% in our genetic linkage map, respectively. This is the first linkage map of Lentinula edodes using SSR markers, which provides essential information for quantitative trait analyses and improvement of genome assembly.

Robust and Cooperative Localization for Underwater Sensor Networks in the Existence of Malicious Anchors.

Precise and robust localization in three-dimensional underwater sensor networks is still an important research problem. This problem is particularly challenging if there are some malicious anchors among ordinary anchor nodes that will broadcast their locations falsely and deliberately. In this paper, we study how to self-localize large teams of underwater sensor nodes under the condition that some malicious anchor nodes mixed with ordinary anchors. Due to malicious characteristic of some deliberate anchor nodes, an iterative and cooperative 3D-localization algorithm for underwater sensor networks in the existence of malicious anchors is proposed in this paper. The proposed robust localization algorithm takes advantage of distributed reputation voting method within 1-Hop neighboring reference nodes to detect and eliminate malicious anchor nodes. Moreover, one kind of Minimum Mean Squared Error estimation based iterative localization method is applied to determine accurate location information. Additionally, we analyze and prove that our localization algorithm would have a bounded error when the number of malicious anchors is smaller than a certain threshold. Extensive simulation results are provided to demonstrate performance improvements comparing to traditional Minimum Mean Squared Error and Attack Resistant Minimum Mean Squared Error based localization methods in terms of localization accuracy and coverage ratio.

Overexpression of the saccharopine dehydrogenase gene improves lysine biosynthesis in Flammulina velutipes.

Saccharopine dehydrogenase (EC 1.5.1.7) regulates the last step of fungal lysine biosynthesis. The gene (Fvsdh) encoding saccharopine dehydrogenase was identified and cloned from the whole genome of Flammulina velutipes. The genomic DNA of Fvsdh is 1257 bp, comprising three introns and four exons. The full-length complementary DNA of Fvsdh comprises 1107 bp with a deduced amino acid sequence of 368 residues. A 1,000-bp promoter sequence containing the TATA box, CAAT box, and several putative cis-acting elements was also identified. The results of tissue expression analysis showed that the expression level of the Fvsdh gene was higher in the pileus than in the stipe whether in the elongation or maturation stage. Further research showed that the lysine contents were 3.03 and 2.95 mg/g in maturation-pileus and elongation-pileus, respectively. In contrast, the lysine contents were 2.49 and 2.07 mg/g in elongation-stipe and maturation-stipe, respectively. To study the function of Fvsdh, we overexpressed Fvsdh in F. velutipes and found that Fvsdh gene expression was increased from 1.1- to 3-fold in randomly selected transgenic strains. The lysine contents were also increased from 1.12- to 1.3-fold in these five transformants, except for strain T3, in which the lysine contents were the same as the control. These results indicate that the expression of the Fvsdh gene can affect the lysine content of F. velutipes.

Metabonomics analysis of serum from rats given long-term and low-level cadmium by ultra-performance liquid chromatography-mass spectrometry.

1. This study evaluated the toxicity of chronic exposure to low-level cadmium (Cd) in rats using ultra-performance liquid chromatography-mass spectrometry (UPLC-MS). Forty male Sprague-Dawley rats were randomly assigned to four groups, namely, the control group, low-dose group (0.13 mg/kg·bw), middle-dose group (0.8 mg/kg·bw) and high-dose group (4.89 mg/kg·bw). The rats continuously received CdCl2 via drinking water for 24 weeks. Serum samples were collected for metabonomics analysis. The data generated from the UPLC-MS was analysed using principal components analysis (PCA) and partial least-squares discriminant analysis (PLS-DA). PLS-DA model with satisfactory explanatory and predictive ability is capable of discriminating the treatment groups from the control group. 2. Finally, the 10 metabolites were identified and showed significant changes in some treatment groups compared with that in the control group (p < 0.0167 or p < 0.003). Exposure to Cd resulted in increased intensities of lysophosphatidic acid (P-16:0e/0:0), glycocholic acid, bicyclo-prostaglandin E2, lithocholyltaurine, sulfolithocholylglycine, lysophosphatidylethanolamine (20:5/0:0) and lysophosphatidylcholine (20:0), as well as decreased intensities of 3-indolepropionic acid, phosphatidylcholine (18:4/18:0) and 15S-hydroxyeicosatrienoic acid in rat serum. 3. Results suggest that exposure to Cd can cause disturbances in the lipid metabolism, amino acid metabolism, nervous system, antioxidant defence system, liver and kidney function.

Smooth 3D Dubins Curves Based Mobile Data Gathering in Sparse Underwater Sensor Networks.

Sensory data collection is one of the most important concerns in underwater sensor networks (USNs). Because full connectivity cannot be guaranteed, mobile data gathering with autonomous underwater vehicles (AUVs) is widely used in sparse three-dimensional (3D) USNs to solve energy-imbalance problems between different sensor nodes. AUVs with relatively abundant energy and storage can collect sensory data from one sensor node to transmit to another node, so as to avoid energy-intensive multi-hop transmission. As a result, movement control strategy and data collecting path planning for AUVs are very crucial for the performance of data acquisition. This paper proposes a smooth 3D Dubins curves based mobile data gathering mechanism to overcome the kinematic nonholonomic constraints of AUVs. The objective of our proposed method is to collect sensory data along smooth 3D Dubins paths, which are interpolated by continuous Bezier curves in the Z-axis from 2D Dubins curves. Extensive simulation results verify that the proposed method has a more efficient performance in terms of path smoothness and energy consumption; thus it is very suitable for mobile data collection in 3D underwater sensor networks.

What influenced the lesion patterns and hemodynamic characteristics in patients with internal carotid artery stenosis? A retrospective study.

OBJECTIVES: This study aimed to explore the dynamic changes of lesion patterns and hemodynamic characteristics in patients with internal carotid artery stenosis (ICAS). PATIENTS AND METHODS: Patients who had suffered an acute ischemic stroke in the distribution of ipsilateral ICAS were included. Computed tomography (CT) and transcranial doppler ultrasound (TCD) were conducted to evaluate the degree of ICAS and the hemodynamic characteristics of the intracranial and extracranial arteries. RESULT: A total of 424 patients were included in the study. With the aggravation of ICAS, blood velocity in ipsilateral ICA was increased, while blood flow in the ipsilateral middle cerebral artery (MCA) was decreased. In the same degree of ICAS, patients with opened communicating arteries showed relatively higher blood perfusion in MCA compared with those without communicating arteries. In the average stage of ICAS, small lesions (D=0-1.5cm), middle lesions (1.5cm3.0cm) commonly existed. The number of small and large lesions significantly increased when the blood flow of ipsilateral MCA decreased. In the same degree of stenosis, the number of small lesions and large lesions, and the total area of all lesions, evidently increased with the decrease of ipsilateral MCA blood velocity. CONCLUSION: Hypoperfusion is an independent risk factor for ischemic lesions in patients with ICAS. Whether or not the communicating arteries are open influences the blood flow of the intracranial arteries. TCD was a convenient and rapid tool to assess intracranial perfusion and vascular compensatory status.

Comparison of intestinal permeability and p-glycoprotein effects on the intestinal absorption of enantiomers of 2-(2-hydroxypropanamido) benzoic acid in rats.

The purpose of this study was to compare intestinal permeability between enantiomers of 2-(2-hydroxypropanamido) benzoic acid ((R)-/(S)-HPABA), a marine-derived antiinflammatory drug, using an in situ single-pass intestinal perfusion (SPIP) model in rats. Concentrations, isolated regions of small intestine, and p-glycoprotein (P-gp) inhibitor were performed to investigate their influences on the intestinal absorption of (R)-/(S)-HPABA. In addition, a molecular docking method was performed to illustrate our prediction. The absorption rate coefficients (Ka ) and permeability values (Peff ) of (R)-/(S)-HPABA were calculated. The permeability of (S)-HPABA was significantly (P < 0.01) higher than that of (R)-HPABA in jejunum, and ileum permeability of (R)-/(S)-HPABA appeared best in ileum; the investigated concentrations ranged from 20 to 80 µg/mL, Ka and Peff values of (R)-/(S)-HPABA increased linearly; in the presence of P-gp inhibitor (verapamil), Peff values of two enantiomers were increased significantly; and the effect of P-gp on absorption of (R)-HPABA is stronger than that of (S)-HPABA in ileum segment. Based on these results, carrier-mediated transport or passive transport combined with carrier-mediated transport seems to be the mechanism for intestinal absorption of (R)-/(S)-HPABA, and (R)-/(S)-HPABA may be recognized as the P-gp substrate. In addition, the intestinal permeability of (S)-HPABA is higher than that of (R)-HPABA.

Relationship between CHOP/GADD153 and unstable human carotid atherosclerotic plaque.

BACKGROUND AND AIMS: The signaling protein C/EBP homologous protein (CHOP) and corresponding growth-arrest-and-DNA-damage-inducible gene 153 (GADD153) is associated with endoplasmic reticulum stress (ERS), which can lead to apoptosis. Our study aims to elucidate the role of CHOP/GADD153 in unstable atherosclerotic (AS) plaque formation isolated from confounding factors such as diabetes mellitus, primary hyperlipidemia, autoimmune deficiencies/abnormalities, essential hypertension, chronic heart failure, chronic kidney disease, and smoking. MATERIAL AND METHODS: We collected carotid artery tissue samples from patients aged 50-80 years-old who received carotid endarterectomies (CEA) at our institution. We obtained fresh AS plaque samples during CEA and preserved the specimens immediately in the operating room with liquid nitrogen. Samples were categorized as stable or unstable AS plaques according to a six-stage histologic classification. CHOP/GADD153 expression was then examined with immunohistochemistry and reverse transcription polymerase chain reaction (RT-PCR). RESULTS: A total of 32 patients met our inclusion and exclusion criteria, with 24 (75.0%) classified as unstable lesions. The mean optical density ratio normalized to GAPDH for CHOP/GADD153 in stable and unstable groups was 0.357 ± 0.025 and 0.490 ± 0.027, respectively (p < .05). Positive immunostaining of CHOP/GADD153 was found in macrophages and smooth muscle cells of unstable AS plaques with a mean integrated optical density of 0.63 ± 0.03, compared to 0.17 ± 0.05 in the stable group (p < .05). CONCLUSIONS: In conclusion, we were able to show significant elevation of CHOP/GADD153 in unstable plaques independent of other confounding factors that induce ERS.

Task Assignment and Path Planning for Multiple Autonomous Underwater Vehicles Using 3D Dubins Curves †.

This paper investigates the task assignment and path planning problem for multiple AUVs in three dimensional (3D) underwater wireless sensor networks where nonholonomic motion constraints of underwater AUVs in 3D space are considered. The multi-target task assignment and path planning problem is modeled by the Multiple Traveling Sales Person (MTSP) problem and the Genetic Algorithm (GA) is used to solve the MTSP problem with Euclidean distance as the cost function and the Tour Hop Balance (THB) or Tour Length Balance (TLB) constraints as the stop criterion. The resulting tour sequences are mapped to 2D Dubins curves in the X - Y plane, and then interpolated linearly to obtain the Z coordinates. We demonstrate that the linear interpolation fails to achieve G 1 continuity in the 3D Dubins path for multiple targets. Therefore, the interpolated 3D Dubins curves are checked against the AUV dynamics constraint and the ones satisfying the constraint are accepted to finalize the 3D Dubins curve selection. Simulation results demonstrate that the integration of the 3D Dubins curve with the MTSP model is successful and effective for solving the 3D target assignment and path planning problem.

Low-pass whole-genome sequencing in clinical cytogenetics: a validated approach.

PURPOSE: Chromosomal microarray analysis is the gold standard for copy-number variant (CNV) detection in prenatal and postnatal diagnosis. We aimed to determine whether next-generation sequencing (NGS) technology could be an alternative method for CNV detection in routine clinical application. METHODS: Genome-wide CNV analysis (>50 kb) was performed on a multicenter group of 570 patients using a low-coverage whole-genome sequencing pipeline. These samples were referred for chromosomal analysis; CNVs (i.e., pathogenic CNVs, pCNVs) were classified according to the American College of Medical Genetics and Genomics guidelines. RESULTS: Overall, a total of 198 abortuses, 37 stillbirths, 149 prenatal, and 186 postnatal samples were tested. Our approach yielded results in 549 samples (96.3%). In addition to 119 subjects with aneuploidies, 103 pCNVs (74 losses and 29 gains) were identified in 82 samples, giving diagnostic yields of 53.2% (95% confidence interval: 45.8, 60.5), 14.7% (5.0, 31.1), 28.5% (21.1, 36.6), and 30.1% (23.6, 37.3) in each group, respectively. Mosaicism was observed at a level as low as 25%. CONCLUSIONS: Patients with chromosomal diseases or microdeletion/microduplication syndromes were diagnosed using a high-resolution genome-wide method. Our study revealed the potential of NGS to facilitate genetic diagnoses that were not evident in the prenatal and postnatal groups.Genet Med 18 9, 940-948.

Reciprocal expression of p-AMPKa and p-S6 is strongly associated with the prognosis of gastric cancer.

Activation of AMP-activated protein kinase (AMPK) suppressed mammalian target of rapamycin (mTOR) pathway, resulting in impaired cancer cell proliferation. Two cohorts (50 and 1072 cases) of patients with resected gastric adenocarcinoma were enrolled in the study. Immunohistochemical staining for p-AMPKa, p-ACC, p-mTOR, p-S6, and p-4EBP1 was performed on the 50-patient cohort. Tissue microarray blocks containing samples from 1072 patients of Chinese ethnicity were used for the immunohistochemical detection of p-AMPKa and p-S6 levels. p-AMPK and p-ACC were frequently inactivated in both cohorts of gastric cancer samples, while p-mTOR, p-S6, and p-4EBP1 were frequently activated in the small cohort of gastric cancer. However, only levels of p-AMPKa and p-S6 were associated with the overall survival of gastric cancer patients. In the larger 1072-patient cohort, downregulation of p-AMPKa and upregulation of p-S6 were associated with tumor progression and were independent predictors of survival after resection of primary gastric cancer. Therefore, reciprocal expression of p-AMPKa and p-S6 may be promising prognostic biomarkers in patients with gastric cancer.

Anesthetic constituents of Zanthoxylum bungeanum Maxim.: A pharmacokinetic study.

A sensitive and selective ultra high performance liquid chromatography with tandem mass spectrometry method was established and validated for the simultaneous determination of hydroxy-α-sanshool, hydroxy-ß-sanshool, and hydroxy-γ-sanshool in rat plasma after the subcutaneous and intravenous administration of an extract of the pericarp of Zanthoxylum bungeanum Maxim. Piperine was used as the internal standard. The analytes were extracted from rat plasma by liquid-liquid extraction with ethyl acetate and separated on a Thermo Hypersil GOLD C18 column (2.1 mm × 50 mm, 1.9 µm) with a gradient elution system at a flow rate of 0.4 mL/min. The mobile phase consisted of acetonitrile/0.05% formic acid in water and the total analysis time was 4 min. Positive electrospray ionization was performed using multiple reaction monitoring mode for the analytes. The calibration curves of the three analytes were linear over the tested concentration range. The intra- and interday precision was no more than 13.6%. Extraction recovery, matrix effect, and stability were satisfactory in rat plasma. The developed and validated method was suitable for the quantification of hydroxy-α-sanshool, hydroxy-ß-sanshool, and hydroxy-γ-sanshool and successfully applied to a pharmacokinetic study of these analytes after subcutaneous and intravenous administration to rats.