Analysis of the improved mechanism of Rhodobacter sphaeroides VK-2-3 coenzyme Q10 by reverse metabolic engineering.

Coenzyme Q10 (CoQ10) is an essential medicinal ingredient. In this study, we obtained a high-yielding mutant strain of CoQ10, VK-2-3, by subjecting R. sphaeroides V-0 (V-0) to a 12C6+ heavy ion beam and high-voltage prick electric field treatment. To investigate the mutation mechanism, the complete genomes of VK-2-3 and V-0 were sequenced. Collinearity analysis revealed that the nicotinamide adenine dinucleotide-dependent dehydrogenase (NAD) gene underwent rearrangement in the VK-2-3 genome. The NAD gene was overexpressed and silenced in V-0, and this construct was named RS.NAD and RS.ΔNAD. The results showed that the titers of CoQ10 in the RS.NAD and RS.ΔNAD increased and decreased by 16.00 and 33.92%, respectively, compared to those in V-0, and these differences were significant. Our results revealed the mechanism by which the VK-2-3 CoQ10 yield increases through reverse metabolic engineering, providing insights for genetic breeding and mechanistic analysis.

Silkworm Cocoon: Dual Functions as a Traditional Chinese Medicine and the Raw Material of Promising Biocompatible Carriers.

The silkworm cocoon (SC), both as a traditional Chinese medicine and as the raw material for biocompatible carriers, has been extensively used in the medical and biomedical fields. This review elaborates on the multiple functions of SC, with an in-depth analysis of its chemical composition, biological activities, as well as its applications in modern medicine. The primary chemical components of SC include silk fibroin (SF), silk sericin (SS), and other flavonoid-like bioactive compounds demonstrating various biological effects. These include hypoglycemic, cardioprotective, hypolipidemic, anti-inflammatory, antioxidant, and antimicrobial actions, which highlight its potential therapeutic benefits. Furthermore, the review explores the applications of silk-derived materials in drug delivery systems, tissue engineering, regenerative medicine, and in vitro diagnostics. It also highlights the progression of SC from laboratory research to clinical trials, emphasizing the safety and efficacy of SC-based materials across multiple medical domains. Moreover, we discuss the market products developed from silk proteins, illustrating the transition from traditional uses to contemporary medical applications. This review provides support in understanding the current research status of SC and the further development and application of its derived products.

Interannual variations in ozone pollution with a dipole structure over Eastern China associated with springtime thermal forcing over the Tibetan Plateau.

The Tibetan Plateau (TP) is essential in modulating climate change in downstream Eastern China (EC). As a meteorology-sensitive pollutant, changes in ozone (O3) in connection with the TP have received limited attention. In this study, using climate analysis of the China High Air Pollutants O3 product and ERA5 reanalysis data of meteorology for 1980-2020, the effect of springtime TP thermal forcing on the warm season (April-September) O3 pollution over EC was investigated. The strong TP thermal effect significantly modulates the interannual variations in O3 pollution with a dipole pattern over EC, inducing more O3 pollution in northern EC regions and alleviating O3 pollution in the southern regions. In northern (southern) EC, strong TP thermal forcing triggers a significant anomalous high (low) pressure center accompanied by anticyclonic (cyclonic) anomalies, resulting in decreased (increased) total cloud cover, increased (reduced) surface downward solar radiation and air temperature, which are conducive to the anomalous increase (decrease) in surface O3 concentrations. Moreover, the key sources of springtime thermal forcing over the TP influence the major O3 pollution regions over southern and northern EC with an inverse pattern, depending on their locations and orientations to the large topography of the TP. This research reveals an important driving factor for the dipole interannual variation in O3 pollution over EC, providing a new prospect for the effect of the TP on atmospheric environmental change.

Inversion of 137Cs emissions following the fukushima accident with adaptive release recovery for temporal absences of observations.

Temporal absences in observation records lead to release losses during the source term inversions of atmospheric radionuclide emissions. Consequently, objectively-estimated source terms for the Fukushima accident contain fewer release details and present large discrepancies when compared with the expert-judged one. This paper describes an objective method that can adaptively recover the missing releases caused by the temporal absences of observations. The proposed method assumes that the accident releases of radionuclides are piecewise-constant and comprise both peaks and constant releases. The missing releases are adaptively recovered as either peaks or constant releases by minimizing the total variation of the estimated source term. The proposed method is applied to the Fukushima accident and evaluated against regional airborne and deposited 137Cs observations. The results demonstrate that this method effectively recovers the missing releases, producing a source term that matches the timing of both on-site gamma dose rate peaks and accident events. The retrieved source term improves the simulation of air concentrations and reproduces most of the deposition patterns. This is the first time that an objective method has independently reproduced the details in the expert-judged one for the Fukushima accident.

Empagliflozin Ameliorates Diabetic Cardiomyopathy via Attenuating Oxidative Stress and Improving Mitochondrial Function.

Diabetic cardiomyopathy (DCM) is considered to be a critical contributor to the development of heart failure. Empagliflozin (EMPA), a sodium-glucose cotransporter 2 inhibitor, has been shown to prevent cardiovascular events and reduce the incidence of heart failure in randomized clinical trials. However, the mechanism of how EMPA prevents DCM is poorly understood. To study the potential mechanisms involved in the therapeutic effects of EMPA, we assessed the protective effects of EMPA on myocardial injury in type 2 diabetic db/db mice and H9C2 cardiomyocytes. 9-10-week-old male db/db mice were treated with EMPA (10 mg/kg) via oral gavage daily for 20 weeks. Afterward, cardiac function of treated mice was evaluated by echocardiography, and pathological changes in heart tissues were determined by histopathological examination and western blot assay. EMPA markedly reduced blood glucose levels, improved insulin tolerance, and enhanced insulin sensitivity of db/db mice. In addition, EMPA significantly prevented cardiac dysfunction, inhibited cardiac hypertrophy and fibrosis, and reduced glycogen deposition in heart tissues. Furthermore, EMPA improved diabetes-induced oxidative stress and mitochondrial dysfunction in both heart tissues of db/db mice and palmitate exposed H9C2 cells. EMPA significantly increased the expression of nuclear factor erythroid 2-related factor 2 (Nrf2) and its downstream genetic targets in cardiac tissue of type 2 diabetic db/db mice and H9C2 cells. EMPA also downregulated the expression of mitochondrial fission-related proteins and upregulated the expression of mitochondrial fusion-related proteins. Collectively, these findings indicate that EMPA may prevent DCM via attenuating oxidative stress and improving mitochondrial function in heart tissue.

Analysis of Gene Co-Expression Network to Identify the Role of CD8 + T Cell Infiltration-Related Biomarkers in High-Grade Glioma.

BACKGROUND: High-grade glioma is a type of heterogeneous lethal brain tumor most common in adults. At present, immune checkpoint inhibitors (ICIs) are being considered for first-line therapeutics for malignant GBM. Nonetheless, molecular markers for malignant GBM are unavailable at present. As a result, it is important to explore molecular markers related to immunity for GBM. MATERIALS AND METHODS: The present study adopted a deconvolution algorithm for quantifying immunocyte composition and measuring gene expression, and used weighted gene co-expression network analysis (WGCNA) to analyze GBM expression data obtained from Gene Expression Omnibus (GEO), Chinese Glioma Genome Atlas (CGGA), and the Cancer Genome Atlas Glioblastoma Multiforme (TCGA-GBM) databases. Thereafter, key CD8+ T cell infiltration-related genes and modules were identified, and database analysis was conducted to verify the therapeutic and immune features of the selected genes. RESULTS: From this study, CD8+ T cell-related modules were identified. By using consistent clustering analysis, two panels of genes (red and green) with the highest correlation with CD8+ T cells infiltration were used to construct high-, low-expression groups, silent and/or mixed group of T cell infiltrations. In the high and low CD8+ T cell infiltration groups, a total of 535 differential genes were obtained, of which ten genes (RPS5, RPS6, FAU, RPS19, RPS23, RPS15A, RPS29, RPS14, RPS16, RPS27A) were identified through protein-protein interactions and co-expression network analysis. Post Cox regression and Kaplan-Meier (K-M) survival analysis, RPS5, RPS6, and RPS16 were selected as candidate prognostic biomarkers related to CD8+ T cells. CONCLUSION: The three associated genes RPS5, RPS6, and RPS16 were markedly related to degree of T cell infiltration and immune-related activated. We identified their potential biomarkers and therapeutic targets associated with the extent of CD8+ T cell infiltration in GBM.

Application of Ausubel cognitive assimilation theory in teaching/learning medical biochemistry and molecular biology.

"Didn't understand or not fully understand" is the most common situation that students experience when they learn medical biochemistry and molecular biology (MBMB). Therefore, the primary task of teaching MBMB is to help students overcome the difficulties in "conceptual understanding." The core of the solution to these problems lies in the acquisition and application of cognitive skills. The key to applying Ausubel cognitive assimilation theory (A's CAT) is to expand the knowledge capacity of students' cognitive structure through the purposeful "production" and employment of advanced organizers, with which students can replace "rote learning" with "meaningful learning" and ultimately develop an interest in MBMB courses. The results showed that the application of A's CAT did not only encourage the students to use cognitive skills to learn and understand MBMB knowledge and overcome the barrier to learning that results from the characteristics of the MBMB course, but also expanded and deepened the MBMB course contents. As a branch of learning theory, A's CAT provides practical methods for the several hypotheses of learning theory. Among these, the advanced organizer strategy realizes the organization and reorganization of new knowledge; the progressive differentiation strategy reduces the difficulty of complex and abstract knowledge; the implementation of the advanced organizer strategy and integrative reconciliation strategy forms the concept map. Taken together, the application of A's CAT includes cognitive strategy and metacognitive strategy when the cognitive process is conducted in a meaningful way. More importantly, application of A's CAT guides students to participate in active learning and builds "scaffolding" for the teaching/learning MBMB of student-centered goals.

[Urban Aerosol Hygroscopicity During Haze Weather].

The hygroscopicity of aerosols has an important influence on atmospheric visibility and is one of the main causes of haze pollution. Based on observations of the aerosol hygroscopic growth factor (GF), water soluble inorganic ions, and organic carbon/elemental carbon (OC/EC) data during haze weather from April 17 to May 21, in 2014, the hygroscopic properties of aerosols and corresponding effects on haze in Nanjing were analyzed. The results showed that the distribution of GF was bimodal and varied from 1.12 to 1.64. With the increase of particle size, the average hygroscopic growth factor (GFmean) changed less and the standard deviation of wettability (σ) increased gradually; meanwhile, the degree of external mixing of chemical components increased gradually. The hygroscopicity of aerosol particles in the day was better than that at night, but the mixing degree was weaker than that at night; in non-haze weather, the hygroscopicity of aerosol particles was stronger and the degree of external mixing was higher, while the hygroscopicity and mixing degree of haze particles showed opposite trends. With the increase of haze levels, the hygroscopicity of aerosol particles grew weaker and the degree of external mixing decreased further. Relative humidity can have a significant impact on the chemical components of aerosols and their hygroscopic capacity. Under a low humidity background, the main chemical components of aerosols included NH4+, NO3-, SO42-, OC, and EC, and the content of OC/EC in aerosols during haze days was more abundant; in haze weather with low relative humidity, abundant organic matter was the main reason for the decrease of the moisture absorption capacity of small-scale aerosols. The level of relative humidity in the haze weather was also an important factor affecting the hygroscopic capacity of aerosols. The contents of (NH4)2SO4, OC, and insoluble substances in aerosols were the highest, followed by NH4NO3. The contents of these chemical components showed obvious diurnal variation characteristics, which resulted in significant diurnal variation of the hygroscopicity of the aerosols. κchem calculated by the chemical composition and κmean acquired by observations using H-TDMA showed good consistency, and the correlation coefficient was 0.8903. In haze weather, the correlation between them was further enhanced. Therefore, the major chemical components of aerosols could be used to predict the hygroscopic properties of aerosols.

A Vision Based Detection Method for Narrow Butt Joints and a Robotic Seam Tracking System.

Automatic joint detection is of vital importance for the teaching of robots before welding and the seam tracking during welding. For narrow butt joints, the traditional structured light method may be ineffective, and many existing detection methods designed for narrow butt joints can only detect their 2D position. However, for butt joints with narrow gaps and 3D trajectories, their 3D position and orientation of the workpiece surface are required. In this paper, a vision based detection method for narrow butt joints is proposed. A crosshair laser is projected onto the workpiece surface and an auxiliary light source is used to illuminate the workpiece surface continuously. Then, images with an appropriate grayscale distribution are grabbed with the auto exposure function of the camera. The 3D position of the joint and the normal vector of the workpiece surface are calculated by the combination of the 2D and 3D information in the images. In addition, the detection method is applied in a robotic seam tracking system for GTAW (gas tungsten arc welding). Different filtering methods are used to smooth the detection results, and compared with the moving average method, the Kalman filter can reduce the dithering of the robot and improve the tracking accuracy significantly.

Targeting Smox Is Neuroprotective and Ameliorates Brain Inflammation in Cerebral Ischemia/Reperfusion Rats.

Spermine oxidase (Smox) is a member of the polyamine oxidases and has been demonstrated to be involved in ischemic brain damage. In this study, we found that Smox expression was increased in a rat middle cerebral artery occlusion (MCAO) model and in cultured primary neurons after oxygen-glucose deprivation and reoxygenation (OGD/R). Smox downregulation by the adeno-associated virus RNA interference system significantly reduced the MCAO-induced brain infarct volume and neurological deficits and decreased neuronal apoptosis and inflammatory reactions. In addition, significant microglial activation and increased IL-6 and TNF-α expression were observed in microglia treated with supernatant from neurons after OGD/R. However, a significant reduction in microglial activation as well as IL-6 and TNF-α expression was observed in microglia treated with supernatant from Smox downregulated neurons after OGD/R. Therefore, the results indicated that Smox is an important mediator of cerebral ischemia injury and may be a therapeutic target for cerebral ischemia patients.

[Characteristics of Key Size Spectrum of PM2.5 Affecting Winter Haze Pollution in Taiyuan].

PM2.5 is generally considered as a main pollutant causing the formation of haze. Based on meteorological parameters, aerosol distribution, and PM monitoring data in Taiyuan during November and December 2016, the characteristics of the key size spectrum of PM2.5 affecting haze were discussed. During the observation period, haze was frequent and serious. Heavy haze time accounts for 25.35% of the total haze time. Haze events occurred frequently when the relative humidity was greater than 80% and wind speed was less than 1.5 m·s-1, especially for severe haze. Mild and moderate level haze occurred frequently when the relative humidity was less than 80% and greater than 40% and when wind speed was less than 1.5 m·s-1. Slight haze mainly occurred when the relative humidity was 20%-40% and the wind speed was 1.25-2.55 m·s-1. The average mass concentration of PM2.5 was 209.45 µg·m-3, which was three times the level during non-haze events. With an increase in the haze level, the mass concentration of PM2.5 and the ratio of PM2.5/PM10 increased. PM1 was the key particle size affecting haze in the low humidity environment. PM0.5 was the key particle size that affects slight haze, mild haze, and moderate haze in the high humidity environment, while PM1 was the key particle size that affects heavy haze. The contribution of surface concentration to visibility decreased with high humidity, but the particle size increased by moisture absorption leading to an increase in the extinction efficiency factor, which compensated for the lack of surface concentration. The increase in the particle size parameter was an important factor for PM2.5 affecting the haze pollution with high humidity.

Surface design and preparation of multi-functional magnetic nanoparticles for cancer cell targeting, therapy, and imaging.

Recently, theranostic candidates based on superparamagnetic iron oxide nanoparticles (SPIONs) providing the combination of therapy and diagnosis have become one of the most promising system in cancer research. However, poor stability, premature drug release, lack of specific tumor cell targeting, and complicated multi-step synthesis processes still hinder them for potential clinical applications. In this research, the multi-functional magnetic nanoparticles (MNPs-DOX) were prepared via a simple assembly process for targeted delivery of doxorubicin (DOX) and enhanced magnetic resonance (MR) imaging detection. Firstly, the multi-functional copolymer coating, polyamidoamine (PAMAM), was designed and synthesized by Michael addition reaction, where N,N-bis(acryloyl)cystamine served as backbone linker, and DOX, dopamine (DA), and polyethylene glycol (PEG) acted as comonomers. The PAMAM was then directly assembled to the surface of SPIONs by the ligand exchange reaction with SPIONs forming the MNPs-DOX. The hydrophilic PEG moieties provide the nanoparticles with colloidal stability and good-dispersity in aqueous solution. Comparing with the quick release of free DOX, the drug release behavior of MNPs-DOX exhibited a sustained drug release. Because the chemical cleavage of disulfide in the polymer backbone, a high cumulative drug release up to 60% in GSH within 48 h was observed, rather than only 26% in PBS (pH 7.4) without GSH. The MR imaging detection experiment showed that the MNPs-DOX had an enhanced T 2 relaxivity of 126 mM-1 S-1 for MR imaging. The results of the cytotoxicity assays showed a remarkable killing effect of cancer cells by MNPs-DOX due to the FA tumor-targeting ligand, comparing with non-targeted drug molecules. All the results showed that the as prepared multi-functional magnetic nanoparticles may serve as a promising theranostic candidate for targeted anticancer drug delivery and efficient detection through MR imaging in medical application.

MicroRNA-133b inhibits hepatocellular carcinoma cell progression by targeting Sirt1.

MicroRNAs (miRNAs) are a class of small non-coding RNAs that function as critical gene regulators by targeting mRNAs for translational repression or degradation. In this study, we showed that the expression level of miR-133b was decreased, while Sirt1 mRNA expression levels were increased in hepatocellular carcinoma (HCC) and cell lines, and we identified Sirt1 as a novel direct target of miR-133b. The over-expression of miR-133b suppressed Sirt1 expression. In addition, miR-133b over-expression resulted in attenuating HCC cell proliferation and invasion together with apoptosis increase in vitro. HepG2 cell transplantation revealed that up-regulation of miR-133b could inhibit HCC tumor genesis in vivo. Forced expression of Sirt1 partly rescued the effect of miR-133b in vitro. Furthermore, our study showed that miR-133b over-expression or Sirt1 down-regulation elevated E-cadherin expression, and repressed glypican-3 (GPC3) and the anti-apoptotic proteins (Bcl-2, Bcl-xL, and Mcl-1) expression. The inhibition of GPC3 expression repressed Bcl-2, Bcl-xL, and Mcl-1 expression, and elevated E-cadherin expression. Moreover, the Sirt1 up-regulation resulted in increases in HCC cell proliferation and invasion together with decreases apoptosis, and increases in the cytosolic accumulation and nuclear translocation of the transcription factor ß-catenin in vitro. But the effect of Sirt1 up-regulation was partly reversed by GPC3 down-regulation in vitro. Taken together, these findings provide insight into the role and mechanism of miR-133b in regulating HCC cell proliferation, invasion and apoptosis via the miR-133b/Sirt1/GPC3/Wnt ß-catenin axis, and miR-133b may serve as a potential therapeutic target in HCC in the future.

Expression profiling suggests a regulatory role of gallbladder in lipid homeostasis.

AIM: To examine expression profile of gallbladder using microarray and to investigate the role of gallbladder in lipid homeostasis. METHODS: 33P-labelled cDNA derived from total RNA of gallbladder tissue was hybridized to a cDNA array representing 17,000 cDNA clusters. Genes with intensities > or =2 and variation <0.33 between two samples were considered as positive signals with subtraction of background chosen from an area where no cDNA was spotted. The average gray level of two gallbladders was adopted to analyze its bioinformatics. Identified target genes were confirmed by touch-down polymerase chain reaction and sequencing. RESULTS: A total of 11 047 genes expressed in normal gallbladder, which was more than that predicted by another author, and the first 10 genes highly expressed (high gray level in hybridization image), e.g. ARPC5 (2 225.88+/-90.46), LOC55972 (2 220.32+/-446.51) and SLC20A2 (1 865.21+/-98.02), were related to the function of smooth muscle contraction and material transport. Meanwhile, 149 lipid-related genes were expressed in the gallbladder, 89 of which were first identified (with gray level in hybridization image), e.g. FASN (11.42+/-2.62), APOD (92.61+/-8.90) and CYP21A2 (246.11+/-42.36), and they were involved in each step of lipid metabolism pathway. In addition, 19 of those 149 genes were gallstone candidate susceptibility genes (with gray level in hybridization image), e.g. HMGCR (10.98+/-0.31), NPC1 (34.88+/-12.12) and NR1H4 (16.8+/-0.65), which were previously thought to be expressed in the liver and/or intestine tissue only. CONCLUSION: Gallbladder expresses 11 047 genes and takes part in lipid homeostasis.