Spleen-based proteogenomics reveals that Escherichia coli infection induces activation of phagosome maturation pathway in chicken.

Avian pathogenic Escherichia coli (APEC) leads to economic losses in poultry industry and is also a threat to human health. Various strategies were used for searching virulence factors, while little is known about the mechanism by which APEC survives in host or is eliminated by host. Thus, chicken colibacillosis model was constructed by intraperitoneally injecting E. coli O78 in this study, then the protein dynamic expression of spleen was characterized at different post-infection times by quantitative proteome. Comparative analysis showed that E. coli induced significant dysregulation at 72 h post infection in spleen tissue. Transcriptomic method was further used to assess the changes of dysregulated proteins at 72 h post infection at the mRNA level. Total 278 protein groups (5.7%) and 2,443 genes (24.4%) were dysregulated, respectively. The upregulated proteins and genes were consistently enriched in phagosome and lysosome pathways, indicating E. coli infection activates phagosome maturation pathway. The matured phagolysosome might kill the invasive E. coli. This study illuminated the genetic dysregulation in chicken spleen at the protein and mRNA levels after E. coli infecting and identified candidate genes for host response to APEC infection.

Value of peripheral blood PCT, CRP, FIB and D-D levels for early diagnosing PTB complicated with bacterial pneumonia / 中国热带医学

@#Abstract: Objective　To investigate the early diagnostic value of peripheral blood procalcitonin (PCT), C-reactive protein (CRP), fibrinogen (FIB) and D-dimer (D-D) levels in patients with pulmonary tuberculosis (PTB) complicated with bacterial pneumonia. Methods　A total of 102 patients who admitted to Department of Tuberculosis of Affiliated Nantong Hospital of Shanghai University from Jan 2021 to May 2022 were enrolled in this study and divided into a group (52 cases) with pulmonary tuberculosis (PTB) patients and a group (50 cases) with PTB patients complicated with bacterial pneumonia. The levels of PCT, CRP, FIB and D-D in the peripheral blood were measured, the differences and correlations in all indicators were compared among two groups. The sensitivity and specificity of these indicators in the early diagnosis of PTB complicated with bacterial pneumonia were analyzed by receiver operating characteristic (ROC) curve. Results　The levels of PCT, CRP, FIB and D-D in the peripheral blood from the PTB complicated with bacterial pneumonia group were 0.06 (0.04, 0.16) ng/mL, 38.00 (3.88, 96.10) mg/L, 4.51 (3.02, 6.07) g/L, and 0.59 (0.34, 1.88) mg/L, respectively, which were significantly higher than corresponding 0.04 (0.03, 0.04) ng/mL, 3.20 (0.84, 7.22) mg/L, 2.96 (2.48, 3.77) g/L, and 0.27 (0.17, 0.36) mg/L in the PTB group (Z=-4.784, -5.233, -3.853, -4.199, all P<0.001). Furthermore, the levels of CRP and FIB in the PTB complicated by bacterial pneumonia group were highly positively correlated (r=0.855, P<0.001). The area under the ROC curve (AUC) of PCT, CRP, FIB and D-D for early diagnosis of PTB complicated with bacterial pneumonia were 0.757, 0.794, 0.747 and 0.764, respectively. In addition, the AUC obtained by simultaneous measurement of PCT, CRP, FIB and D-D was as high as 0.916, and the sensitivity and specificity of diagnosing PTB complicated with bacterial pneumonia were increased to 85.7% and 96.9%, respectively, which were higher than those of individual indicators. Conclusions　Levels of peripheral blood PCT, CRP, FIB, and D-D all show varying degrees of increase in patients with PTB complicated with bacterial pneumonia, and detecting the levels of all four markers, rather than any single marker, can assist in early monitoring whether the tuberculosis patients are complicated with bacterial pneumonia.

Cdc42 Promotes Axonogenesis of Primary Hippocampal Neurons by Inhibiting Glycogen Synthase Kinase-3ß.

BACKGROUND: Progressive axon degeneration is a common pathological feature of neurodegenerative diseases. Cdc42 is a member of the Rho GTPase family that participates in axonogenesis. GSK-3ß is a serine/threonine kinase highly implicated in neuronal development and neurodegeneration. This study aimed to examine whether cdc42 promotes axonogenesis by regulating GSK-3ß activity. METHODS: Hippocampal neurons were isolated from neonatal Sprague-Dawley rats and transfected with designated plasmid vectors to alter the activities of cdc42 and GSK-3ß. LiCl treatment was used to inhibit the GSK-3ß activity in primary neurons. GSK-3ß activity was determined by an enzyme activity assay kit. Immunofluorescence staining was used to detect axons stained with anti-Tau-1 antibody and dendrites stained with anti-MAP2 antibody. RESULTS: Transfection with an active cdc42 mutant (cdc42F28L) decreased the activity of GSK-3ß and induced axonogenesis in primary rat hippocampal neurons, while transfection with a negative cdc42 mutant (cdc42N17) resulted an opposite effect. Moreover, transfection with plasmid vectors carrying wild-type GSK-3ß or a constitutively active GSK3ß mutant (GSK-3ß S9A) increased the activity of GSK-3ß and attenuated axonogenesis of primary hippocampal neurons with excessive cdc42 activity, whereas inhibition of GSK-3ß by LiCl abolished the inhibitory effect of the negative cdc42 mutant on axonogenesis. CONCLUSIONS: This study suggests that cdc42 induces axonogenesis of primary rat hippocampal neurons via inhibiting GSK-3ß activity. These findings support further investigation into the mechanisms of cdc42/GSK-3ß-mediated axonogenesis.

Ac-LysargiNase efficiently helps genome reannotation of Mycolicibacterium smegmatis MC2 155.

Accurate genome annotation, the foundation of life science research in the genome era, is hampered by limited known gene models, nonstandard start codons, and the limited homology of annotated genes in other organisms. LysargiNase mirrors trypsin at the cleavage sites, providing the opportunity to identify peptides other than tryptic peptides. In this study, we used an in-house developed acetylated LysargiNase (Ac-LysargiNase) with higher activity and stability in non-pathogenic Mycolicibacterium smegmatis MC2 155 to supplement the widely used trypsin in proteomic studies. We identified 27,582 peptides from 3844 annotated proteins and 332 novel genome search-specific peptides (GSSPs). Among these GSSPs, 88 peptides were annotated in another M.smegmatis genome database, and 41 were verified as novel peptides by predicted theoretical spectra and their corresponding 15N-labeling spectra. Further analysis revealed that 17 verified GSSPs corrected the N-terminus of the 13 annotated genes. The other 24 verified GSSPs helped identify 17 novel open reading frames (ORFs) missed in previously annotated M. smegmatis genomes. Among these novel ORFs, four relatively small proteins with amino acid residues less than 100 and three were precisely identified with C-terminal peptides. Ac-LysargiNase helps with genome reannotation by identifying new genes and events in proteogenomic studies. SIGNIFICANCE: Correct genomic annotation is vital in the field of life sciences. The nonstandard start codons seriously affect the confirmation of the translation initiation sites (TISs) of an open reading frame (ORF), and unknown structural genes are easily missed in automated gene prediction. Although proteogenomics presents new avenues for validating gene expression and gene structure refinement based on conventional tryptic peptides, determining the TISs and potential encoding genes is complicated. Thus, validation of TISs and encoding ORFs is crucial and urgent. Therefore, we recommend Ac-LysargiNase, a mirror enzyme of trypsin that can identify additional novel peptides for N-terminal correction and ORF identification.

Comparison of Two Different Accesses Single Orifice Percutaneous Endoscopic Surgery in Diagnosis of Ascites of Unknown Origin: A Retrospective Study.

Background: Ascites is a common clinical finding caused by many different diseases, so we developed a technique termed single orifice percutaneous endoscopic surgery (SOPES) which can access peritoneal cavity through the contralateral McBurney's point or umbilicus to seek the underlying causes. In this study, we describe the initial clinical experience of SOPES and compare the application of two accesses. Methods: This is a retrospective study performed between 2007 and 2018. Patients with ascites of unknown origin who underwent these two kinds of SOPES were included. Main outcomes were measured by diagnostic accuracy, complication rate, procedure time, time till stitches removal, length of hospital stay, and hospital cost. Results: 148 patients successfully undergone SOPES via the contralateral McBurney's point (IM group, n = 70) or the umbilicus (UM group, n = 78). 63 patients in the IM group and 71 patients in the UM group reached clear diagnosis (90.0% vs. 91.0%, p = 0.831). The overall complication rate was 5.4%, while the UM group was higher than the IM group (10.3% vs. 0%, p = 0.017). All complications were resolved after medical treatment, and no mortality resulted from this procedure. The procedure time and the time until stitches removal in the UM group were longer than that in the IM group. There were no significant differences in length of hospital stay and hospital cost between the two groups. Conclusions: SOPES, which combines the strength of minimally invasive single orifice incision and flexible angles of examination and instrumentation, is a newly developed flexible endoscopic surgical modality that provides new important clinical valuable in evaluation of ascites of unknown origin. Moreover, SOPES via the contralateral McBurney's point was safer than the umbilicus approach.

Application of Contrast-Enhanced Ultrasound in the Differential Diagnosis of Benign and Malignant Subpleural Pulmonary Lesions.

OBJECTIVE: To explore the clinical value of contrast-enhanced ultrasound (CEUS) in the differential diagnosis of benign and malignant subpleural pulmonary lesions (SPLs). METHODS: Among 959 patients with SPLs who were scheduled to undergo ultrasound-guided puncture in our department between January 2019 and June 2019, 506 patients were included and their B-mode ultrasound and CEUS features, including the lesion's location, size, margin, echo, perfusion pattern of ultrasound contrast agent, degree of enhancement, homogeneity, vascular signs, and necrosis, were retrospectively investigated. All malignant cases were diagnosed by pathology, while benign cases were diagnosed by two respiratory physicians after comprehensive analysis of pathology, etiology, imaging, and clinical symptoms. Statistical differences in these features between the benign and malignant groups were then analyzed. RESULTS: There were 506 cases in this study, including 219 benign cases and 287 malignant cases. Among them, 351 were males and 155 were females, with an average age of 59 ± 16 years. There were statistically significant differences between benign and malignant groups in the perfusion pattern, the degree of enhancement, and vascular signs. The features of the malignant group included local-to-whole perfusion pattern, hypo-enhancement, and curly hair sign, while those of the benign group included a centrifugal perfusion pattern, iso-enhancement and hyper-enhancement, and dendritic sign. There was no statistically significant difference between the two groups in homogeneity and necrosis. CONCLUSIONS: CEUS enhancement mode is different between benign and malignant SPLs, which can provide supplementary information for the differential diagnosis of SPLs in the existing imaging diagnosis.

Deep N-terminomics of Mycobacterium tuberculosis H37Rv extensively correct annotated encoding genes.

Mycobacterium tuberculosis (MTB) is a severe causing agent of tuberculosis (TB). Although H37Rv, the type strain of M. tuberculosis was sequenced in 1998, annotation errors of encoding genes have been frequently reported in hundreds of papers. This phenomenon is particularly severe at the 5' end of the genes. Here, we applied a TMPP [(N-Succinimidyloxycarbonylmethyl) tris (2,4,6-trimethoxyphenyl) phosphonium bromide] labeling combined with StageTip separating strategy on M. tuberculosis H37Rv to characterize the N-terminal start sites of its annotated encoding genes. Totally, 1047 proteins were identified with 2058 TMPP labeled N-terminal peptides from all the 2625 mass spectrometer (MS) sequenced proteins. Comparative genomics analysis allowed the re-annotation of 43 proteins' N-termini in H37Rv and 762 proteins in Mycobacteriaceae. All revised N-termini start sites were distributed in 5'-UTR of annotated genes due to over-annotation of previous N-terminal initiation codon, especially the ATG. In addition, we identified and verified a novel gene Rv1078A in +3 frame different from the annotated gene Rv1078 in +2 frame. Altogether, our findings contribute to the better understanding of N-terminal of H37Rv and other species from Mycobacteriaceae that can assist future studies on biological study.

Quantitative proteomics reveals that dormancy-related proteins mediate the attenuation in mycobacterium strains.

Although members of the Mycobacterium tuberculosis complex (MTBC) exhibit high similarity, they are characterized by differences with respect to virulence, immune response, and transmissibility. To understand the virulence of these bacteria and identify potential novel therapeutic targets, we systemically investigated the total cell protein contents of virulent H37Rv, attenuated H37Ra, and avirulent M. bovis BCG vaccine strains at the log and stationary phases, based on tandem mass tag (TMT) quantitative proteomics. Data analysis revealed that we obtained deep-coverage protein identification and high quantification. Although 272 genetic variations were reported in H37Ra and H37Rv, they showed very little expression difference in log and stationary phase. Quantitative comparison revealed H37Ra and H37Rv had significantly dysregulation in log phase (227) compared with stationary phase (61). While BCG and H37Rv, and BCG and H37Ra showed notable differences in stationary phase (1171 and 1124) with respect to log phase (381 and 414). In the log phase, similar patterns of protein abundance were observed between H37Ra and BCG, whereas a more similar expression pattern was observed between H37Rv and H37Ra in the stationary phase. Bioinformatic analysis revealed that the upregulated proteins detected for H37Rv and H37Ra in log phase were virulence-related factors. In both log and stationary phases, the dysregulated proteins detected for BCG, which have also been identified as M. tuberculosis response proteins under dormancy conditions. We accordingly describe the proteomic profiles of H37Rv, H37Ra, and BCG, which we believe will potentially provide a better understanding of H37Rv pathogenesis, H37Ra attenuation, and BCG immuno protection.

US Contrast Agent Arrival Time Difference Ratio for Benign versus Malignant Subpleural Pulmonary Lesions.

Background US has proven valuable in the diagnosis of subpleural pulmonary lesions (SPLs); however, existing US indicators have limitations. Purpose To propose and validate a revised contrast-enhanced (CE) US indicator for differential diagnosis of benign and malignant SPLs and to compare its performance with existing CE US diagnostic criteria. Materials and Methods This prospective study (Chinese clinical trial registry, ChiCTR1800019828) enrolled patients with SPLs between May 2019 and August 2020. They were divided into a developmental cohort (DC) and a validation cohort (VC). In the DC, the optimal indicator was selected from five CE US indicators. In the VC, the selected indicator was compared with existing CE US diagnostic criteria using the area under the receiver operating characteristic curve (AUC). Pathologic analysis, microbial evidence, and clinical follow-up were used as reference standards for all SPLs. Results A total of 902 participants (DC, 424 participants; VC, 478 participants) with SPLs (mean age, 56 years ± 17; 593 men) were evaluated. The arrival time (AT) difference ratio proved to be the optimal indicator to distinguish benign from malignant SPLs. In the overall (regardless of lesion size), large (vertical diameter >3 cm), and small (vertical diameter ≤3 cm) lesion groups, the cutoff values of the AT difference ratio were 43%, 42%, and 50% and the AUCs obtained from the VC were 0.91 (95% CI: 0.88, 0.93), 0.97 (95% CI: 0.94, 0.98), and 0.77 (95% CI: 0.71, 0.83) respectively, which were higher than those of lesion-lung AT difference greater than 2.5 seconds (0.81 [P < .001], 0.85 [P < .001], and 0.7 [P = .005], respectively), lesion AT greater than 7.5 seconds (0.65 [P < .001], 0.64 [P < .001], and 0.63 [P < .001], respectively), and lesion AT greater than 10 seconds (0.67 [P < .001], 0.68 [P < .001], and 0.64 [P < .001] respectively). Conclusion The US contrast agent arrival time difference ratio enables better differentiation of benign and malignant subpleural lesions when compared with existing diagnostic criteria. Online supplemental material is available for this article. Published under a CC BY 4.0 license.

SARS-CoV-2 infection in the mouse olfactory system.

SARS-CoV-2 infection causes a wide spectrum of clinical manifestations in humans, and olfactory dysfunction is one of the most predictive and common symptoms in COVID-19 patients. However, the underlying mechanism by which SARS-CoV-2 infection leads to olfactory disorders remains elusive. Herein, we demonstrate that intranasal inoculation with SARS-CoV-2 induces robust viral replication in the olfactory epithelium (OE), not the olfactory bulb (OB), resulting in transient olfactory dysfunction in humanized ACE2 (hACE2) mice. The sustentacular cells and Bowman's gland cells in the OE were identified as the major target cells of SARS-CoV-2 before invasion into olfactory sensory neurons (OSNs). Remarkably, SARS-CoV-2 infection triggers massive cell death and immune cell infiltration and directly impairs the uniformity of the OE structure. Combined transcriptomic and quantitative proteomic analyses revealed the induction of antiviral and inflammatory responses, as well as the downregulation of olfactory receptor (OR) genes in the OE from the infected animals. Overall, our mouse model recapitulates olfactory dysfunction in COVID-19 patients and provides critical clues for understanding the physiological basis for extrapulmonary manifestations of COVID-19.

Metformin in Combination with Malvidin Prevents Progression of Non-Alcoholic Fatty Liver Disease via Improving Lipid and Glucose Metabolisms, and Inhibiting Inflammation in Type 2 Diabetes Rats.

BACKGROUND: Non-alcoholic fatty liver disease (NAFLD) is one of the primary causes of chronic liver disease and is closely linked to insulin resistance, type 2 diabetes mellitus (T2DM), and dyslipidemia. However, no effective drug therapies have been approved to treat this disease. The present research aimed to evaluate the therapeutic effects of the combination of oral hypoglycemic drug metformin (MET) and a natural product malvidin (MAL) on hepatic damage in HFD/STZ-induced diabetic rats. METHODS: Sprague-Dawley rats were divided into five groups: normal control group (NC), diabetic control group (DC), DC+MET group, DC+MAL group, and DC+MET+MAL group and treated for eight weeks. Blood and liver tissue samples were collected for metabolic parameters, histological, and RT-qPCR analysis. RESULTS: Our findings indicated that hyperglycemia, insulin resistance, hyperlipidemia, and non-alcoholic fatty liver disease (NAFLD) in diabetic rats were alleviated after oral treatment with MET and MAL, particularly their combination therapy. Besides, the expression of SREBP-1c, ACC, FAS, IL-6, IL-8, and NF-κB mRNA was down-regulated by MET+MAL, and the expression of PPARα, CPT1, and LPL was up-regulated by MET+MAL. CONCLUSION: The evidence of this research indicated that the combination therapy may represent an efficient strategy against NAFLD in T2DM rats via improving lipid and glucose metabolisms, and inhibiting inflammation.

Time-frequency relationship between R&D intensity, globalization, and carbon emissions in G7 countries: evidence from wavelet coherence analysis.

The seven leading industrial countries, called the G7, are becoming a pivotal group to fulfil their emissions-reduction commitments to manage the climate crisis. This paper investigates the relationships between R&D intensity, globalization, and carbon emissions in the context of the G7 countries for the period from 1970 to 2017. Our analysis, which examines these relationships, focuses on the wavelet coherence approach to conduct time-frequency domain analyses. The empirical results show that there is heterogeneity across different time scales and frequencies for R&D intensity, globalization, and carbon emissions within each country. Specifically, R&D intensity and globalization are negatively correlated with carbon emissions for the G7 countries, except Japan, for which they are positive. The long-term correlations between R&D intensity, globalization, and carbon emissions are higher than those in the short- and medium-term periods. In addition, the multiscale connectedness network results reveal that the strongest bidirectional correlations exist between energy consumption, economic growth, and carbon emissions. Our results provide a useful reference for policymakers in the G7 countries to effectively regulate carbon emissions.

Development and Prospective Validation of an Ultrasound Prediction Model for the Differential Diagnosis of Benign and Malignant Subpleural Pulmonary Lesions: A Large Ambispective Cohort Study.

OBJECTIVE: To develop and prospective validate an ultrasound (US) prediction model to differentiate between benign and malignant subpleural pulmonary lesions (SPLs). METHODS: This study was conducted retrospectively from July 2017 to December 2018 (development cohort [DC], n = 592) and prospectively from January to April 2019 (validation cohort [VC], n = 220). A total of 18 parameters of B-mode US and contrast-enhanced US (CEUS) were acquired. Based on the DC, a model was developed using binary logistic regression. Then its discrimination and calibration were verified internally in the DC and externally in the VC, and its diagnostic performance was compared with those of the existing US diagnostic criteria in the two cohorts. The reference criteria were from the comprehensive diagnosis of clinical-radiological-pathological made by two senior respiratory physicians. RESULTS: The model was eventually constructed with 6 parameters: the angle between lesion border and thoracic wall, basic intensity, lung-lesion arrival time difference, ratio of arrival time difference, vascular sign, and non-enhancing region type. In both internal and external validation, the model provided excellent discrimination of benign and malignant SPLs (C-statistic: 0.974 and 0.980 respectively), which is higher than that of "lesion-lung AT difference ≥ 2.5 s" (C-statistic: 0.842 and 0.777 respectively, P <0.001) and "AT ≥ 10 s" (C-statistic: 0.688 and 0.641 respectively, P <0.001) and the calibration curves of the model showed good agreement between actual and predictive malignancy probabilities. As for the diagnosis performance, the sensitivity and specificity of the model [sensitivity: 94.82% (DC) and 92.86% (VC); specificity: 92.42% (DC) and 92.59% (VC)] were higher than those of "lesion-lung AT difference ≥ 2.5 s" [sensitivity: 88.11% (DC) and 80.36% (VC); specificity: 80.30% (DC) and 75.00% (VC)] and "AT ≥ 10 s" [sensitivity: 64.94% (DC) and 61.61% (VC); specificity: 72.73% (DC) and 66.67% (VC)]. CONCLUSION: The prediction model integrating multiple parameters of B-mode US and CEUS can accurately predict the malignancy probability, so as to effectively differentiate between benign and malignant SPLs, and has better diagnostic performance than the existing US diagnostic criteria. CLINICAL TRIAL REGISTRATION: www.chictr.org.cn, identifier ChiCTR1800019828.

Contrast-Enhanced Ultrasound of the Pleural Cavity: A Method to Locate Pleural Catheters and Identify Fibrous Septa.

We aimed to explore the value of contrast-enhanced ultrasound (CEUS) of the pleural cavity in locating catheters and identifying fibrous septa and to compare CEUS with multiple existing methods. We included 304 participants whose pleural effusion could not continue to be drained and compared the catheter-localization capabilities of empirical diagnosis, B-mode ultrasound with normal saline and CEUS, with computed tomography as the reference standard. CEUS performed the best (accuracy, 100%; sensitivity, 100%; specificity, 100%), followed by B-mode ultrasound with normal saline (accuracy, 77.78%; sensitivity, 62.5%; specificity, 100%), and finally empirical diagnosis (accuracy, 54.17%; sensitivity, 66.67%; specificity, 33.33%). The capabilities of CEUS and computed tomography to identify fibrous septa were evaluated, with B-mode ultrasound as the reference, and CEUS (accuracy, 100%; sensitivity, 100%; specificity, 100%) was superior to computed tomography (accuracy, 82.41%; sensitivity, 26.09%; specificity, 97.65%). Overall, CEUS can accurately locate catheters and identify fibrous septa, with performance superior to existing methods.

Cdc42 Facilitates Axonogenesis by Enhancing Microtubule Stabilization in Primary Hippocampal Neurons.

The establishment of polarity is an essential process in early neuronal development. Cdc42, a GTPase of the Rho family, is a key regulator of cytoskeletal dynamics and neuronal polarity. However, the mechanisms underlying the action of cdc42 in regulating axonogenesis have not been elucidated. Here, we expressed wild-type cdc42, a constitutively active cdc42 mutant (cdc42F28L) and a dominant negative cdc42 mutant (cdc42N17), respectively, in the primary hippocampal neurons to alter the activity of cdc42. We found that cdc42 activities were paralleled with the capacities to promote axonogenesis in the cultured neurons. Cdc42 also enhanced microtubule stability in the cultured neurons. Pharmacologically stabilizing microtubules significantly abrogated the defective axonogenesis induced by cdc42 inhibition. Moreover, cdc42 promoted the dephosphorylation of collapsing response mediator protein-2 (CRMP-2) at Thr514 by increasing GSK-3ß phosphorylation at Ser9 in the cultured neurons. These findings suggest that cdc42 may facilitate axonogenesis by promoting microtubule stabilization in rat primary hippocampal neurons.

The direct and indirect effects of democracy on carbon dioxide emissions in BRICS countries: evidence from panel quantile regression.

This paper illustrates the direct and indirect effects of democracy on CO2 emissions in the BRICS countries (Brazil, Russia, India, China, and South Africa) from 1992 to 2018. In view of the distribution heterogeneity of CO2 emissions, the panel quantile regression model is especially used to explore the nexus among different variables. Furthermore, in order to predict the trends of CO2 emissions in different countries, we also estimate the kernel density function of CO2 emissions in the BRICS countries by the quantile-fitted values. The results indicate that the direct impact of democracy on carbon dioxide emissions is significantly negative and great at high-emission countries. Although the indirect effect of democracy is positive in China and negative in Brazil and South Africa, the total effect of democracy on CO2 emissions remains negative in all BRICS countries. The estimation of kernel density function shows that the distribution of CO2 emissions in each country is gradually concentrated. Moreover, there is an environmental Kuznets curve depicting the linkage of urbanization and carbon dioxide emissions in Brazil and South Africa. These findings further highlight that the impact of democracy on high-emission and low-emission countries should be taken into account in policymaking to achieve sustainable developments.

The heterogeneous effect of driving factors on carbon emission intensity in the Chinese transport sector: Evidence from dynamic panel quantile regression.

The transport sector is becoming a key sector for China to accomplish its targets for reducing carbon emission intensity (CEI). Identifying the dominant factors driving CEI of the transport sector is important for CEI mitigation. This paper applied dynamic panel quantile regression to explore the effect of driving factors on CEI in the Chinese transport sector at the provincial level during 2000-2016. The empirical findings indicate that economic growth has a positive influence on CEI at low quantiles, whereas this effect is the opposite at high quantiles. Further, the findings show an inverted U-shaped pattern between economic growth and CEI at low quantiles, which validates the Environmental Kuznets Curve hypothesis in low-CEI provinces. Energy intensity positively influences CEI, with the greatest impact occurring at higher quantiles. Among the lowest CEI provinces, private vehicles and cargo turnover appear to contribute to CEI, and a positive impact of urbanization exists, except at the 5th and 30th quantiles. In conclusion, policy implications for effectively promoting the CEI abatement in the transport sector are discussed.

Classification of Gan Dan Shi Re Pattern and Gan Shen Yin Xu Pattern in Patients with Hepatitis B Cirrhosis Using Metabonomics.

OBJECTIVE: This study aimed to analyze the differential metabolites and their metabolic pathways from the serum of patients with hepatitis B cirrhosis, with two typical patterns of Gan Dan Shi Re (GDSR) and Gan Shen Yin Xu (GSYX) based on the theory of traditional Chinese medicine (TCM). It also investigated the variation in the internal material basis for the two types of patterns and provided an objective basis for classifying TCM patterns using metabolomic techniques. METHODS: The serum samples taken from 111 qualified patients (40 GDSR cases, 41 GSYX cases, and 30 Latent Pattern (LP) cases with no obvious pattern characters) and 60 healthy volunteers were tested to identify the differential substances relevant to hepatitis B cirrhosis and the two typical TCM patterns under the gas chromatography-time-of-flight mass spectrometry platform. The relevant metabolic pathways of differential substances were analyzed using multidimensional statistical analysis. RESULTS: After excluding the influence of LP groups, six common substances were found in GDSR and GSYX patterns, which were mainly involved in the metabolic pathways of glycine, serine, threonine, and phenylalanine. Eight specific metabolites involved in the metabolic pathways of linoleic, glycine, threonine, and serine existed in the two patterns. CONCLUSIONS: The data points on the metabolic spectrum were found to be well distributed among the differential substances between the two typical TCM patterns of patients with hepatitis B cirrhosis using metabolomic techniques. The differential expression of these substances between GDSR and GSYX patterns provided an important objective basis for the scientific nature of TCM pattern classification at the metabolic level.

The heterogeneous effects of urbanization and income inequality on CO2 emissions in BRICS economies: evidence from panel quantile regression.

This paper empirically examines the effects of urbanization and income inequality on CO2 emissions in the BRICS economies (i.e., Brazil, Russia, India, China, and South Africa) during the periods 1994-2013. The method we used is the panel quantile regression, which takes into account the unobserved individual heterogeneity and distributional heterogeneity. Our empirical results indicate that urbanization has a significant and negative impact on carbon emissions, except in the 80th, 90th, and 95th quantiles. We also quantitatively investigate the direct and indirect effect of urbanization on carbon emissions, and the results show that we may underestimate urbanization's effect on carbon emissions if we ignore its indirect effect. In addition, in middle- and high-emission countries, income inequality has a significant and positive impact on carbon emissions. The results of our study indicate that in the BRICS economies, there is an inverted U-shaped environmental Kuznets curve (EKC) between the GDP per capita and carbon emissions. The conclusions of this study have important policy implications for policymakers. Policymakers should try to narrow the income gap between the rich and the poor to improve environmental quality; the BRICS economies can speed up urbanization to reduce carbon emissions, but they must improve energy efficiency and use clean energy to the greatest extent in the process.

Construction of a recombinant pIRES2-EGFP-ARTS plasmid and its effect on LX-2 cells.

The inhibition of the activation of hepatic stellate cells (HSCs) and the induction of their apoptosis have been investigated as potential strategies to counteract the development and progression of liver fibrosis. Previous research has suggested that apoptosis­related protein in the transforming growth factor­ß signaling pathway (ARTS) may serve a significant role in numerous cell types; however, little is known regarding its roles in HSCs. Total RNA was extracted from LX­2 cells, and the human full­length ARTS gene was obtained by reverse transcription­polymerase chain reaction and inserted into the pIRES2­EGFP cloning vector. Subsequently, the recombinant pIRES2­EGFP­ARTS plasmid was transfected into LX­2 cells by FuGENE 6 transfection reagent, and the expression of ARTS was detected by western blotting and fluorescent microscopy. In addition, the effects of pIRES2­EGFP­ARTS on the activation, apoptosis, viability and migration of LX­2 cells were assessed by western blot analysis, TUNEL staining, an MTT assay, and scratch and Transwell assays, respectively. The present results demonstrated that the pIRES2­EGFP­ARTS vector expressing human ARTS was successfully constructed, and the overexpression of ARTS contributed to enhance the apoptosis and inhibit the activation of human LX­2 HSCs. The present findings suggested that ARTS overexpression may have potential as a novel therapeutic strategy to reverse hepatic fibrosis.