SB431542 partially inhibits high glucose-induced EMT by restoring mitochondrial homeostasis in RPE cells.

BACKGROUND: The epithelial-mesenchymal transition (EMT) of retinal pigment epithelial (RPE) cells participated in the development of retinal fibrosis. SB431542 is a small molecule inhibitor with inhibitory effects on the ALK4, ALK5 and ALK7. Our study aimed to explore the effect of SB431542 on the EMT of RPE cells and to provide new ideas for the treatment of retinal fibrosis. METHODS: We performed fundus fluorescein angiography, optical coherence tomography and hematoxylin-eosin staining in vivo to observe the effect of SB431542 on choroidal neovascularization (CNV)-induced retinopathy. The proliferation, migration, cytoskeleton, adhesion, reactive oxygen species (ROS), mitochondrial morphology and membrane potential of RPE cells were observed in vitro through fluorescein diacetate staining, Cell Counting Kit-8 experiment, wound healing assay, phalloidin staining, immunofluorescence, MitoSOX, DCFH-DA, MitoTracker and JC-10 staining. Western blot, reverse transcription quantitative and immunofluorescence were used to detect the expression of EMT-related markers, pERK1/2, pGSK3ß and ß-catenin. RESULTS: SB431542 significantly alleviated retinopathy in the CNV model. The proliferation, migration and adhesion in RPE cells decreased to a certain extent in SB431542 treatment. SB431542 partially normalized the structure of RPE cells. The expression levels of E-cadherin increased, while the expression levels of laminin and N-cadherin decreased with SB431542 treatment. SB431542 reduced the production of total ROS, mitochondrial SOX and recovered the mitochondrial membrane potential to a certain degree. In addition, our study showed that SB431542 downregulated the phosphorylation of ERK1/2, GSK3ß and the expression of ß-catenin. CONCLUSION: SB431542 improved EMT in RPE cells by maintaining mitochondrial homeostasis via the ERK1/2 and GSK3ß/ß-catenin pathways. Video Abstract SB431542 inhibits EMT in RPE cells under high glucose conditions.

Trimester-specific effect of maternal co-exposure to organophosphate esters and phthalates on preschooler cognitive development: The moderating role of gestational vitamin D status.

Organophosphate esters (OPEs) and phthalate acid esters (PAEs) are prevalent endocrine-disrupting chemicals (EDCs). Humans are often exposed to OPEs and PAEs simultaneously through multiple routes. Given that fetal stage is a critical period for neurodevelopment, it is necessary to know whether gestational co-exposure to OPEs and PAEs affects fetal neurodevelopment. However, accessible epidemiological studies are limited. The present study included 2, 120 pregnant women from the Ma'anshan Birth Cohort (MABC) study. The concentrations of tris (2-chloroethyl) phosphate (TCEP), 6 OPE metabolites and 7 PAE metabolites were measured in the first, second and third trimester using ultra-performance liquid chromatography-tandem mass spectrometry (LC-MS). Cognitive development of preschooler was assessed based on the Wechsler Preschool and Primary Scale of Intelligence-Fourth Edition (WPPSI-IV) of the Chinese version. Generalized estimating equations (GEEs), restricted cubic spline (RCS) and generalized additive models (GAMs) were employed to explore the associations between individual OPE exposure and preschooler cognitive development. The quantile-based g-computation (QGC) method was used to estimate the joint effect of PAEs and OPEs exposure on cognitive development. GEEs revealed significant adverse associations between diphenyl phosphate (DPHP) (ß: -0.58, 95% CI: -1.14, -0.01), bis (2-butoxyethyl) phosphate(BBOEP) (ß: -0.44, 95% CI: -0.85, -0.02), bis(1-chloro-2-propyl) phosphate (BCIPP) (ß: -0.81, 95%CI: -1.43, -0.20) and full-scale intelligence quotient (FSIQ) in the first trimester; additionally, TCEP and bis(2-ethylhexyl) phosphate (BEHP) in the second trimester, as well as DPHP in the third trimester, were negatively associated with cognitive development. Through the QGC analyses, mixture exposure in the first trimester was negatively associated with FSIQ scores (ß: -1.70, 95% CI: -3.06, -0.34), mono-butyl phthalate (MBP), BCIPP, and DPHP might be the dominant contributors after controlling for other OPEs and PAEs congeners. Additionally, the effect of OPEs and PAEs mixture on cognitive development might be driven by vitamin D deficiency.

A comparative study on urban waterlogging susceptibility assessment based on multiple data-driven models.

Accurate identification of urban waterlogging areas and assessing waterlogging susceptibility are crucial for preventing and controlling hazards. Data-driven models are utilized to forecast waterlogging areas by establishing intricate relationships between explanatory variables and waterlogging states. This approach tackles the constraints of mechanistic models, which are frequently complex and unable to incorporate socio-economic factors. Previous research predominantly employed single-type data-driven models to predict waterlogging locations and evaluation of their effectiveness. There is a scarcity of comprehensive performance comparisons and uncertainty analyses of different types of models, as well as a lack of interpretability analysis. The chosen study area was the central area of Beijing, which is prone to waterlogging. Given the high manpower, time, and economic costs associated with collecting waterlogging information, the waterlogging point distribution map released by the Beijing Water Affairs Bureau was selected as labeled samples. Twelve factors affecting waterlogging susceptibility were chosen as explanatory variables to construct Random Forest (RF), Support Vector Machine with Radial Basis Function (SVM-RBF), Particle Swarm Optimization-Weakly Labeled Support Vector Machine (PSO-WELLSVM), and Maximum Entropy (MaxEnt). The utilization of diverse single evaluation indicators (such as F-score, Kappa, AUC, etc.) to assess the model performance may yield conflicting results. The Distance between Indices of Simulation and Observation (DISO) was chosen as a comprehensive measure to assess the model's performance in predicting waterlogging points. PSO-WELLSVM exhibited the highest performance with a DISOtest value of 0.63, outperforming MaxEnt (0.78), which excelled in identifying areas highly susceptible to waterlogging, including extremely high susceptibility zones. The SVM-RBF and RF models demonstrated suboptimal performance and exhibited overfitting. The examination of waterlogging susceptibility distribution maps predicted by the four models revealed significant spatial differences due to variations in computational principles and input parameter complexities. The integration of four WSAMs based on logistic regression has been shown to significantly decrease the uncertainty of a single data-driven model and identify the most flood-prone areas. To improve the interpretability of the data model, a geographical detector was incorporated to demonstrate the explanatory capacity of 12 variables and the process of waterlogging. Building Density (BD) exhibits the highest explanatory power in relation to explain waterlogging susceptibility (Q value = 0.202), followed by Distance to Road, Frequency of Heavy Rainstorms (FHR), DEM, etc. The interaction between BD and FHR results in a nonlinear increase in the explanatory power of waterlogging susceptibility. The presence of waterlogging susceptibility risk in the research area can be attributed to the interactions of multiple factors.

Lumbar and neck injuries of occupants in different reclining postures.

PURPOSE: With the increasing level of automation in automobiles, the advent of autonomous vehicles has reduced the tendency of drivers and passengers to focus on the task of driving. The increasing automation in automobiles reduced the drivers' and passengers' focus on driving, which allowed occupants to choose a more relaxed and comfortable sitting position. Meanwhile, the occupant's sitting position went from a frontal, upright position to a more relaxed and reclined one, which resulted in the existing restraint systems cannot to keep occupants safe and secure. This study aimed to determine the effects of different reclining states on occupants' lumbar and neck injuries. METHODS: This is an original research on the field of automotive safety engineering. Occupants in different initial sitting positions (25°, 35°, 45°, and 55°) were adapted to changes in seat back angle and restraint systems and placed in the same frontal impact environment. Neck injury indexes, lumbar axial compression force and acceleration, as well as occupant dynamic response during the impact, were compared in different sitting positions. The injury response and kinematic characteristics of occupants in different reclining positions were analyzed by the control variable method. RESULTS: As the sitting angle increased, the occupant's head acceleration decreased, and the forward-lean angle decreased. Occupants in the standard sitting position had the greatest neck injury, with an Nij of 0.95, and were susceptible to abbreviated injury scale 2+ cervical medullary injuries. As the seatback angle increased, the geometric position of the lumbar spine tended to be horizontal, and the impact load transmitted greater forces to the lumbar spine. The occupant's lumbar injury was greatest in the lying position, with a peak axial compression force on the lumbar region of 5.5 KN, which was 2.3 KN greater than in the standard sitting position. CONCLUSION: The study of occupant lumbar and neck injuries based on different recline states can provide a theoretical basis for optimizing lumbar evaluation indexes, which is conducive to the understanding of the lumbar injury mechanism and the comprehensive consideration of occupant safety protection.

ETNPPL modulates hyperinsulinemia-induced insulin resistance through the SIK1/ROS-mediated inactivation of the PI3K/AKT signaling pathway in hepatocytes.

Hyperinsulinemia is a critical risk factor for the pathogenesis of insulin resistance (IR) in metabolic tissues, including the liver. Ethanolamine phosphate phospholyase (ETNPPL), a newly discovered metabolic enzyme that converts phosphoethanolamine (PEA) to ammonia, inorganic phosphate, and acetaldehyde, is abundantly expressed in liver tissue. Whether it plays a role in the regulation of hyperinsulinemia-induced IR in hepatocytes remains elusive. Here, we established an in vitro hyperinsulinemia-induced IR model in the HepG2 human liver cancer cell line and primary mouse hepatocyte via a high dose of insulin treatment. Next, we overexpressed ETNPPL by using lentivirus-mediated ectopic to investigate the effects of ETNPPL per se on IR without insulin stimulation. To explore the underlying mechanism of ETNPPL mediating hyperinsulinemia-induced IR in HepG2, we performed genome-wide transcriptional analysis using RNA sequencing (RNA-seq) to identify the downstream target gene of ETNPPL. The results showed that ETNPPL expression levels in both mRNA and protein were significantly upregulated in hyperinsulinemia-induced IR in HepG2 and primary mouse hepatocytes. Upon silencing ETNPPL, hyperinsulinemia-induced IR was ameliorated. Under normal conditions without IR in hepatocytes, overexpressing ETNPPL promotes IR, reactive oxygen species (ROS) generation, and AKT inactivation. Transcriptome analysis revealed that salt-inducible kinase 1 (SIK1) is markedly downregulated in the ETNPPL knockdown HepG2 cells. Moreover, disrupting SIK1 prevents ETNPPL-induced ROS accumulation, damage to the PI3K/AKT pathway and IR. Our study reveals that ETNPPL mediates hyperinsulinemia-induced IR through the SIK1/ROS-mediated inactivation of the PI3K/AKT signaling pathway in hepatocyte cells. Targeting ETNPPL may present a potential strategy for hyperinsulinemia-associated metabolic disorders such as type 2 diabetes.

[Research on pretreatment methods for cystine determination in milk powder].

OBJECTIVE: To establish a rapid, accurate and safe pretreatment method for the determination of cystine in milk powder. METHODS: Samples were oxidized at 50 ℃ for 10 min after adding performic acid, and then the reaction was terminated with ethanol as oxidation terminator. The reaction solution was concentrate to dryness by vacuum centrifugation at 50 ℃. The residue was hydrolyzed at 145 ℃ for 4 h after adding 15 mL hydrochloric acid(6 mol/L). Quantitatively transfer the hydrolysate to 25 mL volumetric flask, rinsing with water, and dilute to volume. A proper amount of hydrolysate was concentrated to dryness by vacuum centrifugation at 50 ℃, dissolved in sodium citrate buffer, filtered, and determined by amino acid analyzer. RESULTS: A rapid detection method of cystine in milk powder was realized in this study. Cysteic acid were linearly correlated within the set range, and the correlation coefficients were more than 0.999. This method was applied to the determination of milk powder reference material SRM1849A. The result was within the range of values, and the precision was all less than 4%; The detection limit of cystine was 0.001%, and the limit of quantitation was 0.003%. The applicability of this approach was validated by a comparison study with milk powder reference material(SRM 1869). To explore the scalability of the method, fish meal and soybean meal were measured for six times under the above conditions according to the classification of animals and plants. Meanwhile, milk powder, fish meal and soybean meal were also measured according to GB/T 15399-2018 as reference value. The relative deviation of the average values measured by both method were all less than 6%. CONCLUSION: This method is simple and rapid, with good repeatability and high accuracy, and less harm to the operators and experimental equipment. It can be used for the rapid detection of cystine in milk powder, and it also has applicability to other complex samples.

Self-reported and digital-trace measures of computer science students' self-regulated learning in blended course designs.

This study investigated the extent to which self-report and digital-trace measures of students' self-regulated learning in blended course designs align with each other amongst 145 first-year computer science students in a blended "computer systems" course. A self-reported Motivated Strategies for Learning Questionnaire was used to measure students' self-efficacy, intrinsic motivation, test anxiety, and use of self-regulated learning strategies. Frequencies of interactions with six different online learning activities were digital-trace measures of students' online learning interactions. Students' course marks were used to represent their academic performance. SPSS 28 was used to analyse the data. A hierarchical cluster analysis using self-reported measures categorized students as better or poorer self-regulated learners; whereas a hierarchical cluster analysis using digital-trace measures clustered students as more active or less active online learners. One-way ANOVAs showed that: 1) better self-regulated learners had higher frequencies of interactions with three out of six online learning activities than poorer self-regulated learners. 2) More active online learners reported higher self-efficacy, higher intrinsic motivation, and more frequent use of positive self-regulated learning strategies, than less active online learners. Furthermore, a cross-tabulation showed significant (p < .01) but weak association between student clusters identified by self-reported and digital-trace measures, demonstrating self-reported and digital-trace descriptions of students' self-regulated learning experiences were consistent to a limited extent. To help poorer self-regulated learners improve their learning experiences in blended course designs, teachers may invite better self-regulated learners to share how they approach learning in class.

C2CD4A/B variants in the predisposition of lung cancer in the Chinese Han population.

BACKGROUND: The mRNA levels of C2CD4A and C2CD4B were dysregulation in lung cancer (LC). We aimed to evaluate the role of C2CD4A/B variants in LC susceptibility. METHODS: There were 710 cases with LC and 710 healthy controls enrolled in the study. The genotyping of twelve variants in C2CD4A/B was carried out by Agena MassARRAY system. Odds ratios (ORs) were calculated by logistic regression analysis to assess the relationship between these variants and LC predisposition. RESULTS: Rs8037894 (OR = 0.81, p = 0.005), rs7172432 (OR = 0.83, p = 0.013), rs11856307 (OR = 0.86, p = 0.043), and rs1436953 (OR = 0.79, p = 0.002) were related to the reduced risk of LC. Besides, the relation of rs7172432 with LC risk in subjects aged > 60 years was observed. Rs4502156 conferred to the increased LC risk, while rs1436953 was associated with the lower susceptibility to LC among males. Rs731820, rs4502156, rs11071657, rs7172432, and rs11856307 contributed to the predisposition of LC among subjects with BMI > 24 kg/m2, while rs7495253 was associated with an increased risk of LC in subjects with BMI ≤ 24 kg/m2. The increased LC risk was found in rs4502156, while the protective risk effect of rs8037894, rs7172432, rs11856307, and rs1436953 on the occurrence of LC was observed in smokers and non-drinkers. Moreover, rs7495253 and rs7495931 had a higher risk of lymphatic metastasis. Rs1436953 was related to the reduced risk of lung adenocarcinoma, while rs4502156, rs8037894, rs7172432, rs11856307, and rs1436953 were related to the risk of small cell carcinoma. CONCLUSIONS: Our results first display that C2CD4A/B polymorphisms served as protective factors for LC predisposition in a Chinese Han population. These findings could provide new biological insight into the understanding of C2CD4A/B genes on LC pathogenesis.

Explaining medical students' learning outcomes in blended course designs: combining self-reported and observational learning experiences.

Blended course designs have been increasingly adopted in medical education. However, research on the relations between the key aspects of students' learning experience and their learning outcomes often only measures students' self-reported experience, neglecting what they actually do in learning. This study combined both self-reported and observational measures of students' learning experiences and examined the relations between the two sets of measures and their contributions to learning outcomes. Australian medical students were asked to report their approaches to, and perceptions of, learning. The frequency and duration of their interactions with both online formative and summative tasks were observed and recorded. Correlation analyses showed that the learning outcome was positively related to deep approaches to using online technologies and duration of interactions with online summative tasks. The hierarchical multiple regression analysis found that the self-reported approaches and duration of interactions jointly explained the learning outcomes, accounting for 6% of the variance. The study demonstrated the complementary nature of using both self-reported and observational measures of students' learning experiences to explain the learning outcomes in blended course designs.

A novel multi-model fusion framework diagnoses the complex variation characteristics of ecological indicators and quantitatively reveals their driving mechanism.

Systematic analysis of the change law and driving mechanism of ecological indicators (GPP, ET, WUE), as well as the study of maximum threshold of water resources benefit changing with ecological benefit, are important prerequisites for realizing the scientific allocation and efficient utilization of water resources in desert riparian forests. However, previous studies have defects in the detailed description of the change characteristics of ecological indicators. How to accurately diagnose the characteristics of a site, mutation year, pattern (linear, exponential, logarithmic, etc.), duration of change, future change trends of ecological indicators in a desert riparian environment, as well as quantitatively revealing their driving mechanisms, are major scientific problems that need to be solved urgently. In this regard, an ensemble function coupling a logistic function and an asymmetric Gaussian function was creatively adopted, a novel framework was created to integrate the time-series trajectory fitting method and the sensitivity analysis method, and the arid and ecologically fragile Tarim River Basin was taken as a typical area. The results showed that with enhanced water resource management in the Tarim River Basin, GPP, ET, and WUE all showed patterns of increasing change and could be expected to continue to rise or to remain at a high-level stable state. The longest continuous period of GPP change was 15 years, showing that ecological restoration is a long-term process. The years of GPP mutation were consistent with the implementation periods of major measures in the Tarim River Basin (1990, 2001, and 2011), indicating the reliability of this framework. More importantly, when GPP increased to 216.44 gCm-2, the maximum WUE threshold of 0.93 gCm-2mm-1 occurred. This threshold can be used as a reference criterion for efficient utilization of ecological water in the basin. Among the ecological indicators studied, GPP was the most sensitive to environmental change, but GPP, with 80.60% of pixel area, showed a weak memory effect（α < 0.4). Besides, GPP was the most sensitive to the leaf area index (LAI) and had the strongest correlation with it (p < 0.001). Therefore, LAI can be used as the main control factor for judging plant growth. This research can provide important scientific guidance and reference for the analysis of ecological indicator changes and the sustainable utilization of water resources in arid areas.

Dielectric Relaxation and Magnetic Structure of A-Site-Ordered Perovskite Oxide Semiconductor CaCu3Fe2Ta2O12.

A new perovskite oxide semiconductor, CaCu3Fe2Ta2O12, was synthesized through a high-pressure and high-temperature approach. The compound possesses an Im3̅ space group, where it crystallizes to an A-site-ordered but B-site partial ordered quadruple perovskite structure. Spin ordering occurs around 150 K owing to the antiferromagnetic coupling between Fe3+ spins and ferromagnetic coupling between Cu2+ spins. The room-temperature dielectric permittivity of CaCu3Fe2Ta2O12 was measured to be approximately 2500 at 1 kHz. More importantly, isothermal frequency-dielectric spectroscopy demonstrates the existence of two dielectric relaxations. Debye-like relaxation is attributed to charge carriers trapped among the oxygen vacancies at low temperatures and Maxwell-Wagner polarization relaxation at high temperatures. CaCu3Fe2Ta2O12 is a new magnetic semiconductor, where A-site ordering is intercorrelated with second-order Jahn-Teller distortion. These findings offer opportunities to design novel perovskite oxides with attractive magnetic and dielectric properties.

Configurations of collaborations based on learning orientations amongst medical students.

While collaboration is an important and key attribute for medical students in order to prepare them to perform well in health care teams, how to effectively develop and assess such skills is challenging. The current widespread practice of using Likert-scale questionnaire only to measure the quantity of collaboration at course and/or program level appears to be insufficient to provide an evidence-base for what counts desirable collaborative learning experience. Drawing on research into student approaches to learning and social network analysis, this study investigates differences in collaborative learning configurations amongst 217 Australian medical students. Based on students' learning orientations (i.e., 'understanding' and 'reproducing') and their choice of collaborations (i.e., whether to collaborate or not, with whom to collaborate, and mode of collaboration), the analyses found five configurations of collaborations differing in a number of features. The most desirable collaborative experience was a configuration of collaborations formed by students with an 'understanding' orientation. This configuration revealed a strong tendency towards intensive pair work with measurable differences in how easy and effectively they collaborated. The results of the study not only have practical implications for teaching and curriculum design for collaborative learning, but also have significant implications for assessing students' collaborative learning experiences.

Correction to: FastMM: an efficient toolbox for personalized constraint-based metabolic modeling.

An amendment to this paper has been published and can be accessed via the original article.

FastMM: an efficient toolbox for personalized constraint-based metabolic modeling.

BACKGROUND: Constraint-based metabolic modeling has been applied to understand metabolism related disease mechanisms, to predict potential new drug targets and anti-metabolites, and to identify biomarkers of complex diseases. Although the state-of-art modeling toolbox, COBRA 3.0, is powerful, it requires substantial computing time conducting flux balance analysis, knockout analysis, and Markov Chain Monte Carlo (MCMC) sampling, which may limit its application in large scale genome-wide analysis. RESULTS: Here, we rewrote the underlying code of COBRA 3.0 using C/C++, and developed a toolbox, termed FastMM, to effectively conduct constraint-based metabolic modeling. The results showed that FastMM is 2~400 times faster than COBRA 3.0 in performing flux balance analysis and knockout analysis and returns consistent outputs. When applied to MCMC sampling, FastMM is 8 times faster than COBRA 3.0. FastMM is also faster than some efficient metabolic modeling applications, such as Cobrapy and Fast-SL. In addition, we developed a Matlab/Octave interface for fast metabolic modeling. This interface was fully compatible with COBRA 3.0, enabling users to easily perform complex applications for metabolic modeling. For example, users who do not have deep constraint-based metabolic model knowledge can just type one command in Matlab/Octave to perform personalized metabolic modeling. Users can also use the advance and multiple threading parameters for complex metabolic modeling. Thus, we provided an efficient and user-friendly solution to perform large scale genome-wide metabolic modeling. For example, FastMM can be applied to the modeling of individual cancer metabolic profiles of hundreds to thousands of samples in the Cancer Genome Atlas (TCGA). CONCLUSION: FastMM is an efficient and user-friendly toolbox for large-scale personalized constraint-based metabolic modeling. It can serve as a complementary and invaluable improvement to the existing functionalities in COBRA 3.0. FastMM is under GPL license and can be freely available at GitHub site: https://github.com/GonghuaLi/FastMM.

Identification of continuous immunoglobulin G epitopes of Dermatophagoides farinae allergens by peptide microarray immunoassay.

Dermatophagoides farinae, as a common house dust mite species, is one of the main sources of allergens in the world. At present, Dermatophagoides farinae is found to contain more than 30 groups of allergens. These allergens are used for allergen-specific immunotherapy (AIT) of allergic diseases. During the AIT process, immunoglobulin G (IgG) antibodies can block immunoglobulin E (IgE) antibody-induced allergic reactions in the human body. One of the mechanisms may be that IgG and IgE competitively bind to the same allergic protein, so it is necessary to explore the binding sites (epitopes) of IgG antibodies to allergens. In this study, peptide arrays were constructed to react with the serums from patients with allergic asthma to find the IgG epitopes of several allergens including major allergens (Der f 1, 2) and mid-tier allergens (Der f 4, 5, and 7), and then verified by enzyme-linked immunosorbent assay (ELISA) test. Relevant epitopic sequences were located on the tertiary structure of individual allergens, as reconstructed by homology modeling. One IgG epitope of Der f 1 (90-106aa, NVPSELDLRSLRTVTPI), five IgG epitopes of Der f 4 (61-77aa, ERYQPVSYDIHTRSGDE; 193-209aa, FRSDASTHQWPDDLRSI; 226-242aa, HPFIYHETIYYGGNGIN; 271-287aa, LRWLRNFGTEWGLVPSG; 352-368aa, NDWVGPPTDQHGNILSV), and one IgG epitope of Der f 5 (84-101aa, RYNVEIALKSNEILERDL) were identified. IgG epitopes of Der f 2, 7 were not found. There are overlaps between the IgG and IgE epitopes of Der f 1, 4, and 5. These findings not only reflect the practicality of peptide array and ELISA test in the allergen IgG epitope identification, but also provide more information for further understanding of the human immunological changes during AIT and the molecular mechanisms of IgG blocking IgE activity.

Profiling the pattern of human TRB/IGH-CDR3 repertoire in liver transplantation patients via high-throughput sequencing analysis.

Immune processes in liver transplantation remain poorly understood. Acute allograft rejection in liver transplantation is a kind of T cell-mediated inflammatory disease accompanied by inflammatory cell infiltration. However, the effect of acute allograft rejection on the immunological characteristics of TCRs in peripheral blood mononuclear cell is unknown. In this study, we characterized the pattern of the human T cell receptor beta chain (TRB) and immunoglobulin heavy chain (IGH) complementarity-determining region 3 (CDR3) repertoires via high-throughput sequencing in 11 acute allograft rejection (AG) cases, 23 patients with stable allograft liver function (ST) who had liver transplantation performed and 20 healthy controls (HC). The diversity of TRB-CDR3 was significantly reduced in the AG group compared with the ST group and healthy controls (HC). The CDR3 and N-addition length distribution were not significantly different between the AG and ST groups. However, N-addition length distribution was significantly changed compared to HC. It seemed that AG used more short N-additions and healthy people used more long N-additions in TRB-CDR3 repertoire. Our findings suggested that the TRB-CDR3 region of AG had distinctive V gene use compared with that of HC. The characteristics of ST seemed to be in between those of AG and HC although the difference is not significant. Cluster analysis showed that the TRB repertoire could not effectively distinguish AG from ST. This research might give to a better understanding of the immune process of liver transplantation.

Vitamin D supplementation rescues simvastatin induced myopathy in mice via improving mitochondrial cristae shape.

Statin induced myopathy (SIM) is a main deleterious effect leading to the poor treatment compliance, while the preventive or therapeutic treatments are absent. Mounting evidences demonstrated that vitamin D plays a vital role in muscle as a direct modulator. The deficiency of vitamin D was considered as a cause of muscle dysfunction, whereas the supplementation resulted in a remission. However, there is no causal proof that vitamin D supplementation rescues SIM. Here, using the mice model of simvastatin-induced myopathy, we investigated the role of vitamin D supplementation and the mechanisms associated with mitochondria. Results indicated that simvastatin administration (80 mg/kg) impaired skeletal muscle with the increased serum creatine kinase (CK) level and the declined grip strength, which were alleviated by vitamin D supplementation. Moreover, vitamin D supplementation rescued the energy metabolism dysfunction in simvastatin-treated mice gastrocnemius by reducing the abnormal aggregation of muscular glycogen and lactic acid. Mitochondrial homeostasis plays a key role in the process of energy metabolism. Thus, the mitochondrial dysfunction is a mortal damage for the highly energy-requiring tissue. In our study, the mitochondrial cristae observed under transmission electron microscope (TEM) were lytic in simvastatin-treated gastrocnemius. Interestingly, vitamin D supplementation improved the mitochondrial cristae shape by regulating the expression of mitofusin-1/2 (MFN1/2), optic atrophy 1 (OPA1) and dynamin-related protein 1 (Drp1). As expected, the mitochondrial dysfunction and oxidative stress was mitigated by vitamin D supplementation. In conclusion, these findings suggested that moderate vitamin D supplementation rescued simvastatin induced myopathy via improving the mitochondrial cristae shape and function.

Cytomegalovirus Infection Is a Risk Factor in Gastrointestinal Cancer: A Cross-Sectional and Meta-Analysis Study.

BACKGROUND: This study was planned to investigate the association betweenhuman cytomegalovirus (HCMV) infection and gastrointestinal cancer (GIC) risk, by undertaking a meta-analysis and case-control cross-sectional study. SUMMARY: A cross-sectional study analysis of 160 GIC patients and 100 control subjects indicated significantly higher HCMV prevalence in GIC patients based on the HCMV IgM test. However, a similar analysis based on an IgG test revealed no significant relationship. Further meta-analysis of 11 studies, including 1,044 patients and 991 healthy subjects, displayed HCMV infection as an important risk factor for not only colorectal cancer occurrence and development based on a HCMV DNA test, but also for GIC based on a HCMV IgM test. However, the IgG test again displayed no significant relationship between HCMV infection and GIC occurrence. Key Message: Overall, our study revealed that HCMV infection is associated with an increased GIC risk. However, additional studies are warranted to elucidate the molecular mechanism underlying this association.

Ceramide induces MMP-9 expression through JAK2/STAT3 pathway in airway epithelium.

BACKGROUND: Ceramide, a bioactive lipid, plays an essential role in the development of several pulmonary inflammatory diseases. Matrix metallopeptidase 9 (MMP-9) regulates the synthesis and degradation of extracellular matrix, and is associated with airway remodeling and tissue injury. This study was conducted to investigate the effects and underlying mechanisms of ceramide on MMP-9 expression in airway epithelium. METHODS: BEAS-2B cells, normal human bronchial epithelium cell lines, were pretreated with AG490, a selective janus tyrosine kinase 2 (JAK2) inhibitor, or Stattic, a selective signal transducer and activator of transcription 3 (STAT3) inhibitor. The cells were then stimulated with C6-ceramide. The levels of MMP-9 were determined by ELISA and real-time quantitative PCR (RT-qPCR). JAK2, phosphorylated JAK2 (p-JAK2), STAT3, and phosphorylated STAT3 (p-STAT3) expression was examined by Western blotting. BALB/c mice were pretreated with AG490 or Stattic before intratracheally instillated with C6-ceramide. Pathological changes in lung tissues were examined by Hematoxylin and Eosin staining, Periodic-acid Schiff staining, and Masson's trichrome staining. MMP-9, JAK2, p-JAK2, STAT3, and p-STAT3 expression in the lung tissues was examined by Western blotting. RESULTS: The expression of MMP-9, p-JAK2 and p-STAT3 in BEAS-2B cells was significantly increased after the treatment of C6-ceramide. Furthermore, the increased expression of MMP-9 induced by C6-ceramide was inhibited by AG490 and Stattic. Similar results were obtained in the lung tissues of C6-ceramide-exposed mice which were treated with AG490 or Stattic. CONCLUSIONS: Ceramide could up-regulate MMP-9 expression through the activation of the JAK2/STAT3 pathway in airway epithelium. Targeted modulation of the ceramide signaling pathway may offer a potential therapeutic approach for inhibiting MMP-9 expression. This study points to a potentially novel approach to alleviating airway remodeling in inflammatory airway diseases.

Akebia saponin D alleviates hepatic steatosis through BNip3 induced mitophagy.

Akebia Saponin D (ASD) is the most abundant constituent of the rhizome of Dipsacus asper Wall. The prior studies have shown that ASD alleviates hepatic steatosis targeted at the modulation of autophagy and exerts hepatoprotective effects through mitochondria. However, it is still unclear which signal transduction pathway that ASD increase autophagy and protect the mitochondria. The purpose of this paper was to explore the mechanisms through which ASD alleviates hepatic steatosis. ASD significantly reduced lipid accumulation in BRL cells. Furthermore, ASD significantly increased the mitophagy acting as increase the colocalization between mitochondria and punctate EGFP-LC3. ASD treatment increased the expression of BNip3, phospho-AMPK, prevented oleic acid (OA) induced LC3-II and phospho-mTOR expression. These effects were similar to the effects cotreatment with rapamycin. ASD treatment could not attenuate the expression of BNip3 blocked by chloroquine (CQ) or siRNA-mediated knockdown of BNip3. These results suggest that Akebia saponin D alleviates hepatic steatosis targeted at BNip3 mediated mitophagy. Activation of BNip3 via ASD may offer a new strategy for treating NAFLD.