Enhancing Nix-dependent mitophagy relieves AKI by restricting TREM-1-mediated hyperactivation of inflammasome in platelets.

Platelets are highly involved in inflammation and organ injury under pathological conditions. The mitophagy in platelets may restrict hyperactivation of the inflammasome and relieve acute kidney injury (AKI). Cecal ligation puncture (CLP)/LPS-induced AKI Triggering receptor expressed on myeloid cells (TREM-1)-knockout mice models were established. Additionally, septic patients with AKI were also included. TREM-1 expression in platelets and inflammasome activation were examined. Platelet transfer assays were performed to investigate the contribution of platelet TREM-1 to renal injury. Mitophagy was evaluated in the context of inflammation. BNIP3L/Nix knockout mice were used to examine the relationship between platelet mitophagy and inflammatory activation. The results showed that the level of TREM-1 was increased and the platelet inflammasome was hyperactivated in CLP mice and septic patients, and TREM-1 activated platelet inflammasomes. TREM-1 deletion significantly abrogated hyperactivation of the platelet inflammasome and dramatically reduced AKI, whereas ablation of the mitophagy receptor BNIP3L/Nix induced the accumulation of damaged mitochondria and hyperactivation of platelet inflammasomes in CLP mice. BNIP3L/Nix controlled platelet inflammasome activation, and an amplification loop of platelet inflammasome activation and dysfunctional mitochondria controlled sepsis-related AKI. Therefore, targeting TREM-1 and NLRP3/BNIP3L in platelets may represent a novel therapeutic strategy for treating septic AKI.

PET/d-PET (PdP) Pairing for the Design of Dual-Channel Probes.

Design principles of two-channel fluorescence probes are limited. Herein, we report a new principle, i.e., PET/d-PET (PdP) pairing, for the rational design of two-channel probes. Two fluorophores are required in such a PdP-type probe. They mutually quench their fluorescence via PET and d-PET. In the presence of an analyte-of-interest, such a PdP pair is converted into a FRET pair for signaling. The embodiment of such a principle is Rh-TROX, by tethering a rhodamine fluorophore with an ROS-sensitive probe (TotalROX). Fluorescence of both fluorophores in Rh-TROX was quenched as expected. The addition of highly reactive oxidative species led to the recovery of the fluorescence properties of both. The simultaneous fluorescence enhancement in two channels is a viable way to avoid false-positive signals. The new PdP principle could potentially be applied to the development of probes for another range of substrates.

An Efficient Agrobacterium-Mediated Transformation Method for Hybrid Poplar 84K (Populus alba × P. glandulosa) Using Calli as Explants.

A highly efficient Agrobacterium-mediated transformation method is needed for the molecular study of model tree species such as hybrid poplar 84K (Populus alba × P. glandulosa cv. '84K'). In this study, we report a callus-based transformation method that exhibits high efficiency and reproducibility. The optimized callus induction medium (CIM1) induced the development of calli from leaves with high efficiency, and multiple shoots were induced from calli growing on the optimized shoot induction medium (SIM1). Factors affecting the transformation frequency of calli were optimized as follows: Agrobacterium concentration sets at an OD600 of 0.6, Agrobacterium infective suspension with an acetosyringone (AS) concentration of 100 µM, infection time of 15 min, cocultivation duration of 2 days and precultivation duration of 6 days. Using this method, transgenic plants are obtained within approximately 2 months with a transformation frequency greater than 50%. Polymerase chain reaction (PCR), reverse transcription-PCR (RT-PCR) and ß-galactosidase (GUS) histochemical staining analyses confirmed the successful generation of stable transformants. Additionally, the calli from leaves were subcultured and used to obtain new explants; the high transformation efficiency was still maintained in subcultured calli after 6 cycles. This method provides a reference for developing effective transformation protocols for other poplar species.

Analysis of Long Non-Coding RNA Expression Profiles in Non-Small Cell Lung Cancer.

BACKGROUND/AIMS: Long non-coding RNAs (lncRNAs) play an important role in tumorigenesis. However, the role of lncRNA expression in human Non-small cell lung cancer (NSCLC) biology, prognosis and molecular classification remains unknown. METHODS: We established the IncRNA profile in NSCLC by re-annotation of microarrays from the Gene expression omnibus database. Quantitative real-time PCR was used to determine expression of LINC00342. RESULTS: 6066 differentially expressed IncRNAs were identified and we found a novel IncRNA, LINC00342 was significantly up-regulated in NSCLC tissues compared with normal tissues. We confirmed the over-expression of LINC00342 in a cohort of NSCLC patients and found LINC00342 expression level was positively correlated with lymph node metastasis and TNM stages. Furthermore, in a large online database of 1942 NSCLC patients, high expression of LINC00342 indicated poor Overall survival (HR = 1.28, 95% CI: 1.13-1.45) and post progression survival (HR = 1.43, 95% CI: 1.09-1.88). Bioinformatics analyses showed that LINC00342 was co-expressed with different protein-coding genes in NSCLC and normal tissues. Additionally, gene set enrichment analyses found that PTEN and P53 pathways genes were enriched in the groups with higher LINC00342 expression level. By small interfering RNAs mediated silence of LINC00342, proliferation ability was significantly inhibited in lung cancer cell line. CONCLUSION: To summary, our findings indicate that a set of IncRNAs are differentially expressed in NSCLC and we characterized a novel IncRNA, LINC00342 which is significantly up-regulated in NSCLC and could be a prognostic biomarker.

Comparative Proteomic Analysis of Mature Pollen in Triploid and Diploid Populus deltoides.

Ploidy affects plant growth vigor and cell size, but the relative effects of pollen fertility and allergenicity between triploid and diploid have not been systematically examined. Here we performed comparative analyses of fertility, proteome, and abundances of putative allergenic proteins of pollen in triploid poplar 'ZhongHuai1' ('ZH1', triploid) and 'ZhongHuai2' ('ZH2', diploid) generated from the same parents. The mature pollen was sterile in triploid poplar 'ZH1'. By applying two-dimensional gel electrophoresis (2-DE), a total of 72 differentially expressed protein spots (DEPs) were detected in triploid poplar pollen. Among them, 24 upregulated and 43 downregulated proteins were identified in triploid poplar pollen using matrix-assisted laser desorption/ionisation coupled with time of-flight tandem mass spectrometer analysis (MALDI-TOF/TOF MS/MS). The main functions of these DEPs were related with "S-adenosylmethionine metabolism", "actin cytoskeleton organization", or "translational elongation". The infertility of triploid poplar pollen might be related to its abnormal cytoskeletal system. In addition, the abundances of previously identified 28 putative allergenic proteins were compared among three poplar varieties ('ZH1', 'ZH2', and '2KEN8'). Most putative allergenic proteins were downregulated in triploid poplar pollen. This work provides an insight into understanding the protein regulation mechanism of pollen infertility and low allergenicity in triploid poplar, and gives a clue to improving poplar polyploidy breeding and decreasing the pollen allergenicity.

Comparative genomic analysis of Klebsiella pneumonia (LCT-KP214) and a mutant strain (LCT-KP289) obtained after spaceflight.

BACKGROUND: With the development of space science, it is important to analyze the relationship between the space environment and genome variations that might cause phenotypic changes in microbes. Klebsiella pneumoniae is commonly found on the human body and is resistant to multiple drugs. To study space-environment-induced genome variations and drug resistance changes, K. pneumoniae was carried into outer space by the Shenzhou VIII spacecraft. RESULTS: The K. pneumoniae strain LCT-KP289 was selected after spaceflight based on its phenotypic differences compared to the ground-control strain. Analysis of genomic structural variations revealed one inversion, 25 deletions, fifty-nine insertions, two translocations and six translocations with inversions. In addition, 155 and 400 unique genes were observed in LCT-KP214 and LCT-KP289, respectively, including the gene encoding dihydroxyacetone kinase, which generates the ATP and NADH required for microbial growth. Furthermore, a large number of mutant genes were related to transport and metabolism. Phylogenetic analysis revealed that most genes in these two strains had a dN/dS value greater than 1, indicating that the strain diversity increased after spaceflight. Analysis of drug-resistance phenotypes revealed that the K. pneumoniae strain LCT-KP289 was resistant to sulfamethoxazole, whereas the control strain, LCT-KP214, was not; both strains were resistant to benzylpenicillin, ampicillin, lincomycin, vancomycin, chloramphenicol and streptomycin. The sulfamethoxazole resistance may be associated with sequences in Scaffold7 in LCT-KP289, which were not observed in LCT-K214; this scaffold contained the gene sul1. In the strain LCT-KP289, we also observed a drug-resistance integron containing emrE (confers multidrug resistance) and ant (confers resistance to spectinomycin, streptomycin, tobramycin, kanamycin, sisomicin, dibekacin, and gentamicin). The gene ampC (confers resistance to penicillin, cephalosporin-ii and cephalosporin-i) was present near the integron. In addition, 30 and 26 drug-resistance genes were observed in LCT-KP289 and LCT-KP214, respectively. CONCLUSIONS: Comparison of a K. pneumoniae strain obtained after spaceflight with the ground-control strain revealed genome variations and phenotypic changes and elucidated the genomic basis of the acquired drug resistance. These data pave the way for future studies on the effects of spaceflight.

The association of c.1471G>A genetic polymorphism in XRCC1 gene with lung cancer susceptibility in Chinese Han population.

Lung cancer is one of the most spread cancers in the world. The X-ray repair cross-complementing group 1 (XRCC1) gene plays an important role in the development of lung cancer. The objective of this study is to investigate the potential association of XRCC1 genetic polymorphisms with the susceptibility to lung cancer. In totally, 361 lung cancer patients (male, 276; female, 85; mean age, 62.55) and 361 cancer-free controls (male, 253; female, 108; mean age, 61.33) were enrolled in this case-control study. The genotypes of XRCC1 c.1471G>A genetic polymorphism were determined by the created restriction site-polymerase chain reaction (CRS-PCR) and DNA sequencing methods. The influence of XRCC1 gene on the susceptibility to lung cancer was analyzed by the analyses association. Our data indicated that significant differences were found in the frequencies of allelic and genotypic between lung cancer patients and cancer-free controls. The c.1471G>A genetic polymorphism was statistically associated with increased susceptibility to lung cancer [AA vs. GG: odds ratio (OR)=2.75, 95 % confidence interval (CI) = 1.55-4.88, P<0.001; AA vs. GA/GG: OR=2.69, 95 % CI=1.55-4.69, P<0.001; A vs. G: OR=1.37, 95 % CI=1.09-1.71, P=0.007]. The allele A and genotype AA may contribute to the susceptibility to lung cancer. Taken together, these results showed that the functional c.1471G>A genetic polymorphism of XRCC1 was associated with lung cancer susceptibility in the studied population.

TR1 promotes cell proliferation and inhibits apoptosis through cyclin A and CTGF regulation in non-small cell lung cancer.

The Hippo pathway plays a major role in development and organ size control, and its dysregulation contributes to tumorigenesis. WWTR1 is a transcription coactivator acting downstream of the Hippo pathway. Recently, WWTR1 has been reported to be overexpressed in several human cancers including lung cancer. However, the molecular mechanism of WWTR1 regulating lung cancer aggressiveness remains ambiguous. In the present study, we analyzed the expression of WWTR1 in NSCLC cell lines and found that WWTR1 was overexpressed at both the mRNA and protein levels. Knockdown of WWTR1 by siRNA interference in A549 cells significantly inhibited cell proliferation and increased paclitaxel-induced apoptosis. On the other side, WWTR1 overexpression in HBE cell line promoted cell proliferation and inhibited apoptosis. In addition, we found that the decreased proliferation after siRNA treatment was due to cell cycle arrest. Further analysis showed that WWTR1 could induce cyclin A, connective tissue growth factor (CTGF) expression, and inhibit caspase3 cleavage. In conclusion, WWTR1 promotes malignant cell growth and inhibits apoptosis by cyclin A and CTGF regulation.

Inflammatory factor-specific sumoylation regulates NF-κB signalling in glomerular cells from diabetic rats.

OBJECTIVE: The activation of nuclear factor (NF)-κB by cytokines under hyperglycaemic conditions is a potential mechanism for complications in diabetes. We investigated whether small ubiquitin-like modifier 4 (SUMO4) regulates renal NF-κB signalling in diabetic rats. METHODS: Histological changes in kidney were analysed in diabetic GK rats. The expressions of tumour necrosis factor (TNF)-α, NF-κB (p65), IκBα and SUMO4 in renal tissues were examined by immunohistochemistry and Western blotting. Primary cultured glomerular endothelial cells from rats were stimulated by TNF-α or interleukin (IL)-2. RESULTS: The renal expression of TNF-α, NF-κB (p65), IκBα and SUMO4 was significantly higher in diabetic GK rats than in control rats. In control rats, no nuclear translocation was observed for IκBα or NF-κB (p65). However, in diabetic GK rats, translocation of NF-κB (p65) and IκBα into the nucleus was observed, and the expression of SUMO4 and IκBα was up-regulated in the glomerular endothelial cells. SUMO4 was localised in both the cytoplasm and nucleus, while IκBα was predominantly located in the nucleus after stimulation with TNF-α. In contrast, SUMO4 was localised in the nucleus, and increased cytoplasm SUMO4 localisation was found after stimulation with IL-2. CONCLUSIONS: SUMO4 plays a role in regulating NF-κB signalling in glomerular cells. Cytokines have a unique effect in regulating the sumoylation of NF-κB.

The synergistic therapeutic effect of hepatocyte growth factor and granulocyte colony-stimulating factor on pulmonary hypertension in rats.

Pulmonary arterial hypertension (PAH) is characterized by a progressive increase in pulmonary arterial pressure and vascular resistance. Despite advances in therapy for PAH, its treatment and prognosis remain poor. We aimed to investigate whether the transplantation of bone marrow mesenchymal stem cells (MSCs) overexpressing hepatocyte growth factor (HGF), alone or in combination with granulocyte colony-stimulating factor (G-CSF), attenuates the development of experimental monocrotaline (MCT)-induced PAH. Three weeks after MCT administration, rats were divided into the following groups: (1) untreated (PAH); (2) HGF treated; (3) MSCs administered; (4) HGF-MSCs treated; and (5) HGF-MSCs plus G-CSF treated. After 3 weeks, hemodynamic changes, histomorphology, and angiogenesis were evaluated. To elucidate the molecular mechanisms of vascular remodeling and angiogenesis, serum levels of transforming growth factor (TGF)-ß and endothelin-1 (ET-1) were measured, and the gene and protein expression levels of vascular cell adhesion molecule-1 (VCAM-1) and matrix metalloproteinase-9 (MMP-9) were determined. Compared with the PAH, MSC, and G-CSF groups, the HGF and HGF+G-CSF groups exhibited significantly reduced right ventricular hypertrophy and mean pulmonary arterial pressure (P < 0.05). Histologically, vessel muscularization or thickening and collagen deposition were also significantly decreased (P < 0.05). The number of vessels in the HGF+G-CSF group was higher than that in the other groups (P < 0.05). The TGF-ß and ET-1 concentrations in the plasma of pulmonary hypertensive rats were markedly lower in the HGF and HGF+G-CSF groups (P < 0.05). Furthermore, HGF induced the expression of VCAM-1, and HGF treatment together with G-CSF synergistically stimulated MMP-9 expression. Transplanted HGF-MSCs combined with G-CSF potentially offer synergistic therapeutic benefit for the treatment of PAH.

Space mutagenesis of genetically engineered bacteria expressing recombinant human interferon α1b and screening of higher yielding strains.

The aim of this study was to investigate the space mutagenesis of genetically engineered bacteria expressing recombinant human interferon α1b. The genetically engineered bacteria expressing the recombinant interferon α1b were sent into outer space on the Chinese Shenzhou VIII spacecraft. After the 17 day space flight, mutant strains that highly expressed the target gene were identified. After a series of screening of spaceflight-treated bacteria and the quantitative comparison of the mutant strains and original strain, we found five strains that showed a significantly higher production of target proteins, compared with the original strain. Our results support the notion that the outer space environment has unique effects on the mutation breeding of microorganisms, including genetically engineered strains. Mutant strains that highly express the target protein could be obtained through spaceflight-induced mutagenesis.

Glandular papilloma in lung parenchyma.

Solitary respiratory papilloma is a rare epithelial tumour that can be categorized into multiple subtypes. The glandular type (Glandular papilloma, GP) is the rarest. Most GP occurs in the proximal airways and is only rarely found in the lung parenchyma. In this article, we reported a case of GP in lung parenchyma.

Intelligent Rapid Detection Techniques for Low-Content Components in Fruits and Vegetables: A Comprehensive Review.

Fruits and vegetables are an important part of our daily diet and contain low-content components that are crucial for our health. Detecting these components accurately is of paramount significance. However, traditional detection methods face challenges such as complex sample processing, slow detection speed, and the need for highly skilled operators. These limitations fail to meet the growing demand for intelligent and rapid detection of low-content components in fruits and vegetables. In recent years, significant progress has been made in intelligent rapid detection technology, particularly in detecting high-content components in fruits and vegetables. However, the accurate detection of low-content components remains a challenge and has gained considerable attention in current research. This review paper aims to explore and analyze several intelligent rapid detection techniques that have been extensively studied for this purpose. These techniques include near-infrared spectroscopy, Raman spectroscopy, laser-induced breakdown spectroscopy, and terahertz spectroscopy, among others. This paper provides detailed reports and analyses of the application of these methods in detecting low-content components. Furthermore, it offers a prospective exploration of their future development in this field. The goal is to contribute to the enhancement and widespread adoption of technology for detecting low-content components in fruits and vegetables. It is expected that this review will serve as a valuable reference for researchers and practitioners in this area.

Enabling efficient ample-level oxygen evolution on nickel-iron Prussian blue analogue/hydroxide via hierarchical mass transfer channel construction.

Enhancing double-phase mass transfer capability and reducing overpotential at high currents are critical in the oxygen evolution reaction (OER) catalyst design. In this work, nickel-iron layered double hydroxide (NiFe-LDH) loaded on nickel foam (NF) was used as a self-sacrificing template for subsequent growth of nickel-iron Prussian blue (NiFe-PBA) hollow nanocubes on its sheet arrays. The triple-scale porous structure is therefore in-situ constructed in the produced NiFe-PBA@LDH/NF catalyst, where NiFe-PBA nanocubes, NiFe-LDH sheets and NF skeletons provide pores at hundred-nanometers, microns and hundred-microns, respectively. Due to the successful construction of hierarchical mass transfer channels in the catalyst, the overpotential required to deliver 1000 mA cm-2 OER is only 396 mV, which is 80 mV lower than that of NiFe-LDH/NF with a double-scale porous structure, manifesting the importance of the appropriate mass transfer channels, promoting the potential application of the NiFe-PBA@LDH/NF catalyst in industrial-scale electrolysers.

A multi-omic analysis of an Enterococcus faecium mutant reveals specific genetic mutations and dramatic changes in mRNA and protein expression.

BACKGROUND: For a long time, Enterococcus faecium was considered a harmless commensal of the mammalian gastrointestinal (GI) tract and was used as a probiotic in fermented foods. In recent decades, E. faecium has been recognised as an opportunistic pathogen that causes diseases such as neonatal meningitis, urinary tract infections, bacteremia, bacterial endocarditis and diverticulitis. E. faecium could be taken into space with astronauts and exposed to the space environment. Thus, it is necessary to observe the phenotypic and molecular changes of E. faecium after spaceflight. RESULTS: An E. faecium mutant with biochemical features that are different from those of the wild-type strain was obtained from subculture after flight on the SHENZHOU-8 spacecraft. To understand the underlying mechanism causing these changes, the whole genomes of both the mutant and the WT strains were sequenced using Illumina technology. The genomic comparison revealed that dprA, a recombination-mediator gene, and arpU, a gene associated with cell wall growth, were mutated. Comparative transcriptomic and proteomic analyses showed that differentially expressed genes or proteins were involved with replication, recombination, repair, cell wall biogenesis, glycometabolism, lipid metabolism, amino acid metabolism, predicted general function and energy production/conversion. CONCLUSION: This study analysed the comprehensive genomic, transcriptomic and proteomic changes of an E. faecium mutant from subcultures that were loaded on the SHENZHOU-8 spacecraft. The implications of these gene mutations and expression changes and their underlying mechanisms should be investigated in the future. We hope that the current exploration of multiple "-omics" analyses of this E. faecium mutant will provide clues for future studies on this opportunistic pathogen.

[Protective effects of hepatocyte growth factor on hypoxic human pulmonary microvascular endothelial cells].

OBJECTIVE: To investigate the protective effects of hepatocyte growth factor (HGF) on hypoxic human pulmonary microvascular endothelial cells (HPMECs). METHODS: HPMECs were cultured in vitro, and the hypoxic model was established by the physical method. Cells were divided into 4 groups: the control group, the hypoxic group, HGF group, and phytohemagglutinin (PHA) group. The 7(th) generation of HPMECs was evaluated by the method of immunocytochemistry. The persistence rate of HPMECs was measured by MTT assay and the adhesive cells were counted by the microscopy. The expression of intercellular adhesion molecule-1 (ICAM-1) protein was determined by immunofluorescence staining. RESULTS: The adherence percentage of cells significantly decreased after hypoxia, whereas the expression of the ICAM-1 protein was significantly higher in the hypoxia group than in control group (P<0.01). Compared with the hypoxia group, the persistence and adherence percentage of cells in the HGF group significantly increased (P<0.01), whereas the expression of the ICAM-1 protein significantly dropped (P<0.01). In the PHA group, the persistence and adhesion rate were significantly different from those in the hypoxia group and HGF group (P<0.01), and the expression of the ICAM-1 protein increased significantly (P<0.01). CONCLUSION: HGF could inhibit the hypoxic damage of HPMECs by decreasing the persistence and the adhesive capacity of these cells and inducing the expression of ICAM-1.

[Current situation of awareness and use of banned pesticides among vegetable greenhouse farmers in a town of Ledu County, China].

OBJECTIVE: To investigate the awareness and use of banned pesticides among vegetable greenhouse farmers in a town of Ledu County, Qinghai Province, China and to provide a basis for the management and control of banned pesticides. METHODS: Local residents who lived in a town of Ledu County for more than 5 years were selected as subjects in August 2012. A questionnaire survey was conducted in 267 vegetable greenhouse farmers who were selected from five villages of the town by cluster random sampling. RESULTS: Of the 267 subjects, 249 (93.26%) completed and returned the questionnaires. Eighty-seven (34.9%) of the 249 subjects were aware of banned pesticides, and the banned pesticide awareness rate varied significantly among the subjects with different education levels (χ(2) = 11.061, P = 0.011). Only education level entered the regression equation in non-conditional logistic regression analysis. Only five (2.0%) of the 249 subjects knew the banned pesticides as well as the details. All the 249 subjects used banned pesticides to varying degrees in the past 5 years. Only 9.0% (17/189) of the vegetable greenhouse farmers knew the time pesticides were initially banned in China. CONCLUSION: The banned pesticide awareness rate is relatively low in the town of Ledu County, and this rate is influenced by education level. It is needed to spread the knowledge and hazards of banned pesticides.

Computed tomography manifestations of a case of varicella zoster virus (VZV) pneumonia.

Disseminated VZV infections is rare in healthy adults. Several studies have reported VZV reactivation and eruption happens in an immunocompromised host especially after solid organ transplantation. Nonetheless, diffuse bilateral lung VZV infections is also rare. We report a case of disseminated VZV pneumonia after renal transplantation and methylprednisolone treatment. This report highlights the computed tomography manifestations of disseminated VZV pneumonia.

A case of tuberculous pleurisy characterized by bleeding and necrosis documented by medical thoracoscope.

Tuberculous pleurisy is a main cause of pleural effusions. The main histological abnormalities in pleural biopsy of tuberculous pleurisy are caseating granulomas and epithelioid cell granuloma. In our case, chronic inflammation of fibrous tissue with bleeding, necrosis, and exudation were observed during a medical thoracoscopy as manifestations of tuberculous pleurisy.