Evaluation of IFIT3 and ORM1 as Biomarkers for Discriminating Active Tuberculosis from Latent Infection.

OBJECTIVE: Current commercially available immunological tests cannot be used for discriminating active tuberculosis (TB) from latent TB infection. To evaluate the value of biomarker candidates in the diagnosis of active TB, this study aimed to identify differentially expressed genes in peripheral blood mononuclear cells (PBMCs) between patients with active TB and individuals with latent TB infection by transcriptome sequencing. METHODS: The differentially expressed genes in unstimulated PBMCs and in Mycobacterium tuberculosis (Mtb) antigen-stimulated PBMCs from patients with active TB and individuals with latent TB infection were identified by transcriptome sequencing. Selected candidate genes were evaluated in cohorts consisting of 110 patients with TB, 30 individuals with latent TB infections, and 50 healthy controls by quantitative real-time RT-PCR. Receiver operating characteristic (ROC) curve analysis was performed to calculate the diagnostic value of the biomarker candidates. RESULTS: Among the differentially expressed genes in PBMCs without Mtb antigen stimulation, interferon-induced protein with tetratricopeptide repeats 3 (IFIT3) had the highest area under curve (AUC) value (0.918, 95% CI: 0.852-0.984, P<0.0001) in discriminating patients with active TB from individuals with latent TB infection, with a sensitivity of 91.86% and a specificity of 84.00%. In Mtb antigen-stimulated PBMCs, orosomucoid 1 (ORM1) had a high AUC value (0.833, 95% CI: 0.752-0.915, P<0.0001), with a sensitivity of 81.94% and a specificity of 70.00%. CONCLUSION: IFIT3 and ORM1 might be potential biomarkers for discriminating active TB from latent TB infection.

Mesenchymal-epithelial Transition Factor Regulates Monocyte Function during Mycobacterial Infection via Indoleamine 2,3-dioxygenase.

OBJECTIVE: Mycobacterium tuberculosis (Mtb), the causative agent of tuberculosis (TB), causes an estimated 1.6 million human deaths annually, but the pathogenesis of TB remains unclear. Immunity plays a critical role in the onset and outcome of TB. This study aimed to uncover the roles of innate and adaptive immunity in TB. METHODS: The gene expression profiles generated by RNA sequencing from human peripheral blood mononuclear cells (PBMCs) stimulated with or without Mtb strain H37Rv antigens were analyzed. A total of 973 differentially expressed mRNAs were identified. RESULTS: The differentially expressed genes were enriched in innate immunity signaling functions. The mesenchymal-epithelial transition factor (MET) gene was significantly upregulated in CD14+ monocytes. A MET inhibitor improved the uptake of the BCG strain by monocytes and macrophages as well as inhibited the expression of indoleamine 2,3-dioxygenase (IDO). The expression of IDO was increased in PBMCs stimulated with Mtb antigens, and the IDO inhibitor promoted the expression of CD40, CD83, and CD86. CONCLUSION: Our results might provide clues regarding the immunomodulatory mechanisms used by Mtb to evade the host defense system.

Diffusion tensor imaging revealed different pathological processes of white matter hyperintensities.

BACKGROUND: Although increasing evidence showed the correlations between white matter hyperintensities (WMHs) and cognitive impairment, the relationship between them is still modest. Many researchers began to focus on the variation caused by the heterogeneity of WMH. We tried to explore the pathological heterogeneity in WMH by using diffusion tensor imaging (DTI), so as to provide a new insight into the future research. METHODS: Diffusion weighted images (DWIs) of the brain were acquired from 73 patients with WMH and 18 healthy controls, which were then modeled by DTI. We measured fractional anisotropy (FA), mean diffusivity (MD), axial diffusivity (AD), and radial diffusivity (RD) of white matter of the periventricular frontal lobe (pFL), periventricular occipital lobe (pOL), periventricular parietal lobe (pPL) and deep centrum ovales (dCO), and grouped these measures according to the Fazekas scale. Then we compared the DTI metrics of different regions with the same Fazekas scale grade. RESULTS: Significantly lower FA values (all p < 0.001), and higher MD (all p < 0.001) and RD values (all p < 0.001) were associated with WMH observed in the periventricular frontal lobe (pFL) compared to all other regions with the same Fazekas grades. The AD of WMH in the pFL was higher than that of pPL and dCO, but the differences between groups was not as high as of MD and RD, as indicated by the effect size. In the normal control group, DTI metrics between pFL and other regions were not significantly different or less significant different. The difference of DTI metrics of WMH between pPL, pOL and dCO was lower than that of normal white matter, as indicated by the effect size. CONCLUSION: Distinct pathological processes can be revealed by DTI between frontal periventricular WMH and other regions. These processes may represent the effects of severe demyelination within the frontal periventricular WMH.

Turning pig manure into biochar can effectively mitigate antibiotic resistance genes as organic fertilizer.

The composting of fresh manure is an effective way to inactivate pathogens and reduce the levels of antibiotics and some antibiotic resistance genes (ARGs) prior to its application on agricultural land as organic fertilizer. However, some ARGs could still exist and even be enriched after composting. This study investigated whether converting composted pig manure into biochar could reduce the dissemination of ARGs into the soil in comparison with a compost amendment. We performed a pot experiment using pakchoi (Brassica chinensis), with two pig manure-based composts and the biochar derived from composted pig manure, as organic fertilizers. The distributions of the antibiotic resistome, mobile genetic elements (MGEs) and bacterial community composition in soils during cultivation were evaluated by high-throughput qPCR and Illumina sequencing. The total ARGs and MGEs abundance in the biochar-treated soils were significantly lower than those in the compost-amended soils during cultivation. The total ARGs abundance in the biochar-amended soils was similar to that in the control soils during cultivation. Thus, the dissemination of ARGs from animal waste to the environment can be effectively mitigated by converting manure into biochar.

Spontaneous isolated dissection and atherosclerotic plaques of superior mesenteric artery: the vastly different occurrence site suggests the opposite haemodynamic aetiology.

OBJECTIVE: The development of atherosclerotic plaques and spontaneous isolated dissection (SID) of the superior mesenteric artery (SMA) was considered to be related to opposite haemodynamics. The purpose of this study was to compare their occurrence sites and the morphology of the SMA to confirm the haemodynamic aetiologies. METHODS: 57 patients with SID and 64 patients with atherosclerotic plaques were compared about patient characteristics, location of SID and plaque, the distance from lesion to the aortic ostia, SMA branching angle and inlet diameter of the SMA. RESULTS: The location of SID and plaque was very different (p < 0.001). The anterior wall was the most common entry site of SIDSMA (84.0%) but the least frequent origin site of atherosclerotic plaques (7.8%). The posterior, left and right walls were the frequent origin sites of atherosclerotic plaques (total 92.2%) but not for SIDSMA. Most plaques started from the aortic ostia, and their average distance to the aortic ostia was significantly less than the distance from the entry site to the aortic ostia of SIDSMA (p < 0.001). No significant difference was found between SIDSMA and the plaque groups in the branching angle and inlet diameter of the SMA. CONCLUSION: The vastly different sites of SIDSMA and atherosclerotic plaque indicate their opposite haemodynamic aetiology. Advances in knowledge: By comparing the location of the two diseases, we demonstrate their different haemodynamic causes.

Effect of ammonium pyrrolidine dithiocarbamate (PDTC) on NF-κB activation and CYP2E1 content of rats with immunological liver injury.

CONTEXT: Ammonium pyrrolidine dithiocarbamate (PDTC) is a potent inhibitor of nuclear factor-κB (NF-κB). Recent studies have shown that NF-κB plays an essential role in the regulation of genes whose products are involved in the pathogenesis of immunological liver injury. OBJECTIVE: To study the function of NF-κB in immunological liver injury of rat model and its effect on CYP2E1 content and metabolic activity. MATERIALS AND METHODS: The present study investigated the effect of passivating NF-κB activation on CYP2E1 using Bacillus calmette Guérin (BCG)-induced immunological liver injury in Sprague-Dawley rats measured in terms of enzyme levels. The degree of hepatic injury of rats was measured by using biochemical parameters, hepatic tissue pathological changes, and physiological parameters. Protein localization of liver NF-κB was detected by immunohistochemical assay. Western blot analysis was used to detect the protein expression of NF-κB, IκBα, iNOS, and CYP2E1. The content of CYP2E1 of homogenate in the rat liver was detected by ELISA assay and the enzyme kinetics of CYP2E1 probe drug chlorzoxazone was evaluated by high-performance liquid chromatography (HPLC) assay. RESULTS: The results showed that BCG-pretreatment (125 mg/kg) significantly (p < 0.01) increased the weight of liver and spleen (increased by 70% and 248%, respectively), serum levels of alanine transarninase (ALT) and aspartate aminotransferase (AST) (increased by 200% and 75.8%, respectively), the expression of NF-κB and iNOS (increased by 228% and 303%, respectively), and decreased CYP2E1 content and metabolic activity (p < 0.05). Administration of PDTC (50, 100, and 200 mg/kg) reversed above hepatic injury stimulated by BCG in vivo. Moreover, PDTC (ED50: 76 mg/kg) dose dependently inhibited down-regulation of CYP2E1 (p < 0.05). CONCLUSION: Passivation of NF-κB can inhibit the down-regulation of CYP2E1 and iNOS to induce in rat liver tissue with immunological liver injury; NF-κB may be involved in the CYP2E1 regulation through iNOS.

Type II, but not type I, cGMP-dependent protein kinase reverses bFGF-induced proliferation and migration of U251 human glioma cells.

Previous data have shown that the type II cGMP­dependent protein kinase (PKG II) inhibits the EGF­induced MAPK signaling pathway. In order to thoroughly investigate PKG, it is necessary to elucidate the function of another type of PKG, PKG I. The aim of this study was to investigate the possible inhibitory effect of PKG II and PKG I activity on the basic fibroblast growth factor (bFGF)­induced proliferation and migration of U251 human glioma cells and the possible underlying mechanisms. U251 cells were infected with adenoviral constructs encoding cDNA of PKG I (Ad­PKG I) or PKG II (Ad­PKG II) to increase the expression levels of PKG I or PKG II and then treated with 8­Br­cGMP and 8­pCPT­cGMP, respectively, to activate the enzyme. An MTT assay was used to detect the proliferation of the U251 cells. The migration of the U251 cells was analyzed using a Transwell migration assay. Western blot analysis was used to detect the phosphorylation/activation of the fibroblast growth factor receptor (FGFR), MEK and ERK and the nuclear distribution of p-ERK. The results showed that bFGF treatment increased the proliferation and migration of U251 cells, accompanied by increased phosphorylation of FGFR, MEK and ERK. Furthermore, the nuclear distribution of p-ERK increased following bFGF treatment. Increasing the activity of PKG II through infection with Ad-PKG II and stimulation with 8-pCPT-cGMP significantly attenuated the aforementioned effects of the bFGF treatment, while increased PKG I activity did not inhibit the effects of bFGF treatment. These data suggest that increased PKG II activity attenuates bFGF­induced proliferation and migration by inhibiting the MAPK/ERK signaling pathway, whereas PKG I does not.

Endogenous cGMP-dependent protein kinase reverses EGF-induced MAPK/ERK signal transduction through phosphorylation of VASP at Ser239.

In our previous study, we demonstrated that type II cGMP-dependent protein kinase (PKG II) was expressed at lower levels in different human cancer cell lines and that exogenous PKG II inhibited epidermal growth factor (EGF)-induced MAPK/ERK signaling. In order to investigate its functions further in this signaling pathway, it is necessary to elucidate whether endogenous PKG has the same effect or not. This study aimed to investigate the possible inhibitory effect of endogenous PKG activity on EGF-induced MAPK/ERK signal transduction in human lung cancer cells and its mechanism. Human small cell lung carcinoma cells (SCLCs) were treated with the PKG-selective cGMP analog 8-pCPT-cGMP to activate endogenous PKG, EGF and cGMP followed by EGF, respectively. The results showed that increased endogenous PKG activity inhibited the EGF-induced phosphorylation of the epidermal growth factor receptor (EGFR) and the binding between Sos1 and Grb2. In addition, EGF-triggered Ras activation was reversed by increased endogenous PKG activity. While the EGF-induced phosphorylation of MEK and ERK were inhibited by increased endogenous PKG activity, there was a significant increase of phosphorylated vasodilator-stimulated phosphoprotein (p-VASP) at Ser239. Furthermore, we investigated whether endogenous PKG exerted its effects on EGF-induced MAPK/ERK signaling through phosphorylation of VASP at Ser239. Downregulation of the levels of p-VASP Ser239 by point mutation blocked the effects of endogenous PKG on EGF-induced MAPK/ERK signal transduction. The data shown here suggest that endogenous PKG reverses the EGF-induced MAPK/ERK signaling pathway by phosphorylating VASP at Ser239.

[Effects of fungi on co-metabolic degradation of benzo [a] pyrene in droughty red soil].

Simulated bioslurry remediation of PAHs contaminated soil was carried out. Penicillium, Aspergillus niger and white-rot fungus etc. three strains of fungi isolated from petroleum-contaminated soils were inoculated into droughty red soils differently in application rates of phenanthrene and phthalic acid, to investigate their effects of co-metabolic degradation of B[a]P. Results show that in natural soils, some native microorganisms were able to degrade B[a] P and with addition of low molecular weight PAHs-phenanthrene increased degradation rate of B[a] P in the soil. The effect was greater when the application rate of phenanthrene was 100 mg x kg(-1) than 200 mg x kg(-1). But the addition of phthalic acid did not show much effect. In sterilized soils, degradation of B[a]P in soils was hardly observed, and application of co-metabolism has no significant effect. However, inoculation of Penicillium stimulated degradation of B[a]P in all three treatments, i.e., phenanthrene at 100 mg x kg(-1), phenanthrene at 200 mg x kg(-1) and phthalic acid, but the effect of phenanthrene treatment was better than that of phthalic acid treatment. Inoculation of Aspergillus niger also showed similar effect, however, was inhibited by the presence of phenanthrene and phthalic acid in the soil. The degradation ability of white-rot fungus to B[a]P was very poor, but both kinds of phenanthrene concentration and phthalic acid treatments all could promote white-rot fungus to degrade B[a]P in soils, and the effect of phenanthrene was better than that of phthalic acid.

[Evolvement of soil bacterial community in intensively managed Phyllostachys praecox stand and related affecting factors].

By using PCR-DGGE and redundancy analysis (RDA), this paper studied the variations of soil bacterial community structure during the intensive management of Phyllostachys praecox stand, and analyzed the soil factors affecting the diversity of the bacterial community. The results showed that after the transformation of paddy field into P. praecox stand, the Shannon and richness indices of soil bacterial community increased significantly. Long-term intensive management of P. praecox stand altered the community structure of soil bacteria, reflecting in the changes of main bacterial populations and the great decrease after an initial increase of the diversity indices of soil bacterial community. Soil pH, soil total nitrogen, and soil available nitrogen and potassium could explain 76.1% of the total variation of soil bacterial community, and soil pH showed the greatest effect though it was not significant, indicating that the evolvement of soil bacterial community under the long-term intensive management of P. praecox stand was the result of the synergistic effects of several soil factors.

[Spatiotemporal evolvement of soil microbiological characteristics in upland fields with different utilization duration in Cixi, Zhejiang Province].

The microbial number, microbial biomass, and enzymatic activities in five upland soils under agricultural utilization for 50-700 years were determined, with the correlations between soil microbiological characteristics and agricultural utilization duration analyzed. In the meantime, the functional diversity of microbes in soils having been utilized for 50, 100, and 700 years were investigated. The results showed that at the early stage (< 100 years) of agricultural utilization, the number of soil fungi (F) had a slight increase, while the bacterial number (B), B/F ratio, microbial biomass C (C(mic)), microbial biomass N (N(mic)), and the activities of catalase, invertase and urease all decreased markedly. After utilized for more than 100 years, the F decreased significantly, while the B, B/F ratio, C(mic), N(mic), and the activities of test enzymes all tended to increase. During the whole utilization period from 50 to 700 years, the C(mic)/N(mic) ratio had a significant increase with year. The Shannon, Simpson, and McIntosh indices of soil microbial community had the same responses to the agricultural utilization duration as the bacterial number, microbial biomass, and enzymatic activities. All of these indicated that in the upland fields in Cixi of Zhejiang Province, shifts of soil microbial community occurred with increasing agricultural utilization duration, and soil microbiological quality had an overall increase after 100 years agricultural utilization.

Sulfur (S)-induced enhancement of iron plaque formation in the rhizosphere reduces arsenic accumulation in rice (Oryza sativa L.) seedlings.

The effects of two sulfur (S) sources (SO(4)(2-), S(0)), and three rates of S application (0, 30, 120 mgS/kg) on the formation of iron plaque in the rhizosphere, and on the root surface of rice, and As (arsenic) uptake into rice (Oryza sativa L.) were studied in a combined soil-sand culture experiment. Significant differences in As uptake into rice between +S and -S treatments were observed in relation to S sources, and rates of S application. Concentrations of As in rice shoots decreased with increasing rates of S application. The mechanism could be ascribed to sulfur, induced the formation of iron plaque, since concentrations of Fe in iron plaque on quartz sands in the rhizosphere, and on the root surface of rice increased with increasing rates of S application. The results suggest that sulfur fertilization may be important for the development approaches to reducing As accumulation in rice.

[Distribution characteristics and sources identification of PAHs in ancient paddy soil].

Soil samples of current paddy soils, ancient paddy soil/ancient dry land soil, and bottom soils were collected from two sites in Chuodunshan Site of Majiabang Culture (about 6,000a). 15 polycyclic aromatic hydrocarbons (PAHs) were analyzed with HPLC, and their possible sources were identified. The sum of 15 PAHs was 202.9microg x kg(-1) and 207.7microg x kg(-1) in the surface soils from Site A and Site B respectively, which were mainly deposited from atmosphere. In ancient paddy soil from Site A, the total PAHs concentration sharply decreased to 56.0microg x kg(-1), but was still higher than those in ancient dry land soil and bottom soils with the sum of 32.0 approximately 36.9microg x kg(-1). In ancient paddy soil, the concentrations of 2-ring and 3-ring PAHs took a larger portion of 63 percents to the total PAHs, and naphthalene and phenanthrene were the most abundant compounds, while PAHs of more than 4 rings took a small part. The ratios of phenanthrene anthracene and benzo(a) anthracene chrysene and 13C-NMR spectrum of soil organic matter showed that PAHs in ancient paddy soil mainly originated from rice straw burning, biogenesis under reducing conditions may be another source.