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Streptococcus pneumoniae relies on two-component systems (TCSs) to regulate the processes of pathogenicity, osmotic pressure, chemotaxis, and energy metabolism. The TCS01 system of S. pneumoniae is composed of HK01 (histidine kinase) and RR01 (response regulator).

Ex vivo lung perfusion (EVLP) is an excellent platform to apply novel therapeutics, such as gene and cell therapies, before lung transplantation. We investigated the concept of human donor lung engineering during EVLP by combining gene and cell therapies. Premodified cryopreserved mesenchymal stromal cells with augmented anti-inflammatory interleukin-10 production (MSCIL-10) were administered during EVLP to human lungs that had various degrees of underlying lung injury. Cryopreserved MSCIL-10 had excellent viability, and they immediately and efficiently elevated perfusate and lung tissue IL-10 levels during EVLP. However, MSCIL-10 function was compromised by the poor metabolic conditions present in the most damaged lungs. Similarly, exposing cultured MSCIL-10 to poor metabolic, and especially acidic, conditions decreased their IL-10 production. In conclusion, we found that “off-the-shelf” MSCIL-10 therapy of human lungs during EVLP is safe and feasible, and results in rapid IL-10 elevation, and that the acidic target-tissue microenvironment may compromise the efficacy of cell-based therapies., Graphical abstract, Mesenchymal stromal cells with augmented anti-inflammatory IL-10 production were administered to human lungs during ex vivo lung perfusion, a platform that enables application of novel therapeutics to donor lungs before transplantation. IL-10 production was rapid but compromised by lung injury and acidosis, indicating that the local target-tissue microenvironment affects cell-based therapies.

Background: Congenital hypothyroidism is often caused by genetic mutations that impair thyroid hormone (TH) production, resulting in growth and development defects. XB130 (actin filament associated protein 1 like 2) is an adaptor/scaffold protein that plays important roles in cell proliferation, migration, intracellular signal transduction, and tumorigenesis. It is highly expressed in thyrocytes, however, its function in the thyroid remains largely unexplored. Methods: Xb130(−/−) mice and their littermates were studied. Postnatal growth and growth hormone levels were measured, and responses to low or high-iodine diet, and levothyroxine treatment were examined. TH and thyrotropin in the serum and TH in the thyroid glands were quantified. Structure and function of thyrocytes in embryos and postnatal life were studied with histology, immunohistochemistry, immunofluorescence staining, Western blotting, and quantitative reverse transcription polymerase chain reaction. Results: Xb130(−/−) mice exhibited transient growth retardation postnatally, due to congenital hypothyroidism with reduced TH synthesis and secretion, which could be rescued by exogenous thyroxine supplementation. The thyroid glands of Xb130(−/−) mice displayed diminished thyroglobulin iodination and release at both embryonic and early postnatal stages. XB130 was found mainly on the apical membrane of thyroid follicles. Thyroid glands of embryonic and postnatal Xb130(−/−) mice exhibited disorganized apical membrane structure, delayed folliculogenesis, and abnormal formation of thyroid follicle lumina. Conclusion: XB130 critically regulates folliculogenesis by maintaining apical membrane structure and function of thyrocytes, and its deficiency leads to congenital hypothyroidism.

Background Ex vivo lung perfusion (EVLP) is an isolated organ assessment technique that has revolutionized the field of lung transplantation and enabled a safe increase in the number of organs transplanted. The objective of this study was to develop a protein-based assay that would provide a precision medicine approach to lung injury assessment during EVLP. Methods Perfusate samples collected from clinical EVLP cases performed from 2009 to 2019 were separated into development (n=281) and validation (n=57) sets to derive and validate an inflammation score based on IL-6 and IL-8 protein levels in perfusate. The ability of an inflammation score to predict lungs suitable for transplantation and likely to produce excellent recipient outcomes (time on ventilator ≤ three days) was assessed. Inflammation scores were compared to conventional clinical EVLP assessment parameters and associated with outcomes, including primary graft dysfunction and patient care in the ICU. Results An inflammation score accurately predicted the decision to transplant (AUROC 68% [95% CI 62-74]) at the end of EVLP and those transplants associated with short ventilator times (AUROC 73% [95% CI 66-80]). The score identified lungs more likely to develop primary graft dysfunction at 72-hours post-transplant (OR 4.0, p=0.03). A model comprised of the inflammation score and ∆PO2 was able to determine EVLP transplants that were likely to have excellent recipient outcomes, with an accuracy of 87% [95% CI 83-92]. Conclusions The adoption of an inflammation score will improve accuracy of EVLP decision-making and increase confidence of surgical teams to determine lungs that are suitable for transplantation, thereby improving organ utilization rates and patient outcomes.

C64H44Br4N8O8Zn2, monoclinic, P21/c (No. 14), a = 6.6263(5) Å, b = 24.355(2) Å, c = 18.2418(15) Å, β = 96.433(2)°, V = 2925.4(4) Å3, Z = 2, R gt (F) = 0.0400, wR ref (F 2) = 0.1302, T = 296(2) K.

In order to obtain a better fermentation parameter for the production of recombinant Ganoderma lucidum immunomodulatory protein (rFIP-glu), an engineered Pichia pastoris GS115 was investigated on t...

Introduction: Ex vivo lung perfusion (EVLP) is a normothermic platform used to assess donor lungs. Many have studied biomarkers in lung injury, but it is unclear whether samples taken from one location are representative of the organ. Our objective was to investigate the uniformity of cytokine expression in tissue biopsies and in EVLP perfusates from various locations. Methods: In the tissue study, eight donor lungs were partitioned from apex to base. In each lung, three biopsies were taken from the third, sixth, and ninth slices, while two were taken from the lingula and an injury site. In the perfusate study, four samples were taken from four lobes in eight donors during EVLP. Expressions of IL-6, IL-8, IL-10, and IL-1β were measured using qPCR and ELISA. Results: In the tissue study, the mean intra-biopsy equal-variance F-value was 0.53. The median intra-biopsy coefficient of variation (CV) was 18%. In donors without gross focal injury, the mean comparisons of biopsies in each donor showed that the three consistent slices showed no differences and had a CV of 20%, which was similar to the intra-biopsy CV (p=0.80). Both the lingula and injury biopsies demonstrated larger differences from the rest. The median intra-lung CV of perfusates from different lobes was 4.9%. Conclusion: Intra-biopsy variances were consistent across biopsies. Lungs without gross focal injury demonstrated more consistent gene expression. The lingula is not a representative site due to high signal variability. The consistent measurements in EVLP perfusates provided a uniform picture of the inflammation.

C13H9ClO2, monoclinic, P21 (No. 4), a = 6.5465(9) Å, b = 8.0647(11) Å, c = 10.8476(15) Å, β = 105.591(3)°, V = 551.63(13) Å3, Z = 2, R gt (F) = 0.0532, wR ref (F 2) = 0.1507, T = 296(2) K.

To study the effect of combined ultraviolet (UV) chloramine disinfection on viruses in a drinking water supply system, a full-scale experiment was conducted to analyze the distribution, variability, community structure, and hosts of viruses using metagenomics. The results showed that the combined UV chloramine process reduced the number of virus species (6.13%) and gene abundance (51.97%) but did not completely remove the viruses from the water. The United States Environmental Protection Agency (USEPA) report that virus removal efficiencies from water can reach 99%-99.99% based on culturing methods. However, in this study, metagenomic analysis indicated a total virus removal rate of only 93.46%. Therefore, the detection of viruses in water using culturing method cannot reliably detect viruses in drinking water. Caudovirales are the most abundant type of virus in water supply systems and are sensitive to chloramine disinfection.

Application of chemotherapeutic oxaliplatin represses gene transcription through induction of DNA methylation, which may contribute to oxaliplatin-induced chronic pain. Here, Ddr1, which showed an increased methylation in the promoter, was screened from the SRA methylation database (PRJNA587622) after oxaliplatin treatment. qPCR and MeDIP assays verified that oxaliplatin treatment increased the methylation in Ddr1 promoter region and decreased the expression of DDR1 in the neurons of spinal dorsal horn. In addition, overexpression of DDR1 by intraspinal injection of AAV-hSyn-Ddr1 significantly alleviated the mechanical allodynia induced by oxaliplatin. Furthermore, we found that oxaliplatin treatment increased the expression of DNMT3b and ZEB1 in dorsal horn neurons, and promoted the interaction between DNMT3b and ZEB1. Intrathecal injection of ZEB1 siRNA inhibited the enhanced recruitment of DNMT3b and the hypermethylation in Ddr1 promoter induced by oxaliplatin. Finally, ZEB1 siRNA rescued the DDR1 downregulation and mechanical allodynia induced by oxaliplatin. In conclusion, these results suggested that the ZEB1 recruited DNMT3b to the Ddr1 promoter, which induced the DDR1 downregulation and contributed to the oxaliplatin-induced chronic pain.

To investigate the ecological mechanisms of the bacterial response to human disturbance in micro-polluted water ecosystems, we studied the relationship between the environmental factors and bacterial community development in the incoming water and various areas of a reservoir based on an ecological null model. The results showed that the phylogenetic clustering of bacterial communities was more dispersed than expected (the single-sample

XB130 is an adaptor protein that functions as a mediator of multiple tyrosine kinases important for regulating cell proliferation, survival, migration and invasion. Formerly predicted as an oncogene, alterations of its expression are documented in various human cancers. However, the exact role of XB130 in tumorigenesis is unknown. To address its function in skin tumorigenesis, a two-stage dimethylbenzanthracene (DMBA)/12-O-tetradecanoylphorbol 13-acetate (TPA) study was performed on XB130 knockout (KO), heterozygous (HZ) and wild-type (WT) littermate mice. DMBA/TPA-treated XB130 KO and HZ males developed a significantly higher number of epidermal tumors that were notably larger in size than did WT mice. Interestingly, DMBA/TPA-treated female mice did not show any difference in tumor multiplicity regardless of the genotypes. The skin tumor lesions of XB130 KO males were more progressed with an increased frequency of keratoacanthoma. Deficiency of XB130 dramatically increased epidermal tumor cell proliferation. The responses to DMBA and TPA stimuli were also individually investigated to elucidate the mechanistic role of XB130 at different stages of tumorigenesis. DMBA-treated male XB130 KO mice showed compensatory p53-mediated stress response. TPA-treated XB130 KO males demonstrated more skin ulceration with more severe edema, enhanced cell proliferation, accumulation of infiltrating neutrophils and increased production of pro-inflammatory cytokine genes compared with WT mice. Enhanced activities of nuclear factor-kappa B pathway, increased protein expression of metalloproteinase-9 and ERK1/2 phosphorylation were found in these KO mice. These findings demonstrate that XB130 acts as a tumor suppressor in carcinogen-induced skin tumorigenesis that may be mediated through inhibiting inflammation.

Background Increasing evidence indicates that regulated necrosis plays a critical role during cell death caused by ischemia-reperfusion (IR) injury. Necroptosis is one form of regulated necrosis. Necrostatin-1 (Nec-1), an inhibitor of receptor-interacting protein kinase 1 (RIPK1), is known to reduce necroptosis. We investigated the effect of Nec-1 treatment on IR-induced lung injury in a rat lung transplant model. Methods Lewis rats were divided into 4 groups (n = 6 each): (1) Control (no treatment), (2) Donor treatment (D), (3) Recipient treatment (R), and (4) Donor plus Recipient treatment (D+R) groups. Donor lungs were flushed and preserved for 18 hours at 4oC before transplantation. Recipient animals underwent a left single lung transplant. After 2 hours of reperfusion, we assessed the physiologic function, cytokine expression, pathway activation, and the extent of necrosis. Results Pulmonary gas exchange in D+R group was significantly better than in the other 3 groups ( p = 0.003). Lung edema was significantly lower in the D+R group compared with the Control group (p = 0.006). The expression of interleukin-6 in lung tissue and plasma was significantly reduced in the D+R group compared with the Control group (p = 0.036). The percentage of necrotic cells in D+R group was significantly lower than in the Control and D groups (p = 0.01), indicating Nec-1inhibited regulated necrosis. Conclusions The administration of Nec-1 to both donor and recipient improved graft function after lung transplantation through the reduction of necroptosis. The inhibition of regulated necrosis appears to be a promising strategy to attenuate IR lung injury after lung transplantation.

Ischemia-reperfusion (I/R)-induced lung injury undermines lung transplantation (LTx) outcomes by predisposing lung grafts to primary graft dysfunction (PGD). Necrosis is a feature of I/R lung injury. However, regulated necrosis (RN) with specific signaling pathways has not been explored in an LTx setting. In this study, we investigated the role of RN in I/R-induced lung injury. To study I/R-induced cell death, we simulated an LTx procedure using our cell culture model with human lung epithelial (BEAS-2B) cells. After 18 h of cold ischemic time (CIT) followed by reperfusion, caspase-independent cell death, mitochondrial reactive oxygen species production, and mitochondrial membrane permeability were significantly increased. N-acetyl-Leu-Leu-norleucinal (ALLN) (calpain inhibitor) or necrostatin-1 (Nec-1) [receptor interacting serine/threonine kinase 1 (RIPK1) inhibitor] reduced these changes. ALLN altered RIPK1/RIPK3 expression and mixed lineage kinase domain-like (MLKL) phosphorylation, whereas Nec-1 did not change calpain/calpastatin expression. Furthermore, signal transducer and activator of transcription 3 (STAT3) was demonstrated to be downstream of calpain and regulate RIPK3 expression and MLKL phosphorylation during I/R. This calpain-STAT3-RIPK axis induces endoplasmic reticulum stress and mitochondrial calcium dysregulation. LTx patients’ samples demonstrate that RIPK1, MLKL, and STAT3 mRNA expression increased from CIT to reperfusion. Moreover, the expressions of the key proteins are higher in PGD samples than in non-PGD samples. Cell death associated with prolonged lung preservation is mediated by the calpain-STAT3-RIPK axis. Inhibition of RIPK and/or calpain pathways could be an effective therapy in LTx.

Many middle-aged and older adults have more than one chronic health condition. It is therefore important to explore the effectiveness of interventions for multiple chronic conditions. Tai Chi is widely used in China and other countries, and many studies have examined the effect of Tai Chi on anxiety and depression. However, there are no systematic reviews of the effect of Tai Chi on anxiety and depression in various chronic conditions. This systematic review and meta-analysis aimed to evaluate the effects of Tai Chi on anxiety and depression symptoms in four chronic conditions: cancer, stroke, heart failure (HF), and chronic obstructive pulmonary disease (COPD).We searched Chinese and English databases (Cochrane Library, PubMed, Web of Science, China National Knowledge Infrastructure (CNKI), China Science and Technology Journal Database (VIP), Wanfang Database, and Sinomed) from inception to October 2020. Review Manager version 5.2 and Stata version 12.0 were used to perform a systematic review. The quality of the included studies was evaluated using the Cochrane risk of bias tool. The study was registered with the PROSPERO database (number CRD42020209594).Of the 596 studies identified, we included 25 randomized controlled trials involving 1819 participants. Combined analysis of the four diseases showed statistically significant differences between the Tai Chi and control groups for anxiety symptoms (SMD -0.99, 95%CI: -1.5, -0.47; P 0.01) and depressive symptoms (SMD 0.70, 95%CI: -1.01, -0.39; P 0.01). Subgroup analyses showed statistically significant differences between the Tai Chi and control groups for depressive symptoms in stroke (SMD -0.43, 95%CI: -0.67, -0.18; P 0.01) and HF (SMD -0.57, 95%CI: -0.8, -0.33; P 0.01). However, no statistically significant differences were found for depressive symptoms in COPD or cancer. There were statistically significant differences between the Tai Chi and control groups for anxiety symptoms in stroke (SMD -0.60, 95%CI: -0.88, -0.32; P 0.01) and cancer (SMD -0.69, 95%CI: -1.22, -0.17; P 0.01), but not in COPD or HF. Subgroup, sensitivity, meta regression, and publication bias analyses showed high heterogeneity correlated with a single study and study quality. Sensitivity analysis showed that most meta-analysis results had good stability, but those for anxiety symptoms in COPD were unstable; therefore, careful interpretation is required.Tai Chi has a positive effect on anxiety and depression, especially for patients with cancer, stroke, and HF. However, given the weak evidence, this approach is not a substitute for psychiatric treatment.

Objective To investigate the effect of Xiaoyao San on the brain-gut axis in rats exposed to chronic immobilization stress (CIS). Methods Rats were divided into control, model, and treatment groups. The rats belonging to the model and treatment groups were subjected to CIS for 21 consecutive days, during which they were administered Xiaoyao San decoction [3.854 g/(kg·d)] or vehicle by gavage, and their body weight gain, food intake and water intake were monitored. The rats were subsequently subjected to the open field test (OFT) and d -xylose absorption test, and the expression levels of neuropeptides secreted by the hypothalamus and stomach were determined by enzyme-linked immunosorbent assay (ELISA), radioimmune analysis, or real-time fluorescence quantitative polymerase chain reaction. Gastric mucosal morphology was also assessed. Results The model rats exhibited complex brain-gut axis abnormalities following exposure to CIS, abnormalities signified by decreases in food intake, reductions in digestive absorption, decreases in body weight, decreases in the total distances traveled and increases in the time in the central zone during the OFT, gastric mucosal lesion development and decreases in gastrointestinal hormone secretion. These changes were reversed after treatment with Xiaoyao San , which also regulated the secretion of both peripheral (serum and stomach) and central (hypothalamus) brain-gut peptides. Specifically, the levels of neuropeptide Y (NPY) and neuropeptide Y receptor Y5, which are secreted by the hypothalamus and promote digestive function, were increased in the Xiaoyao San -treated group compared with the model group. Furthermore, the levels of pro-opiomelanocortin (POMC) and its receptor, melanocortin-4 receptor (MC4R), which are secreted by the hypothalamus and inhibit digestive function, were significantly decreased in the treatment group compared with the model group. However, the levels of ghrelin (GHRL), gastrin (GAS) and motilin (MTL), which are secreted by the stomach, were significantly increased in the serum and stomach of the treatment group compared with the serum and stomach of the model group following Xiaoyao San treatment ( P vs . the model group). Conclusion Xiaoyao San attenuates CIS-induced gastrointestinal dysregulation by regulating the peptides secreted by both the hypothalamus and the gastrointestinal tract (GIT), suggesting that its effects are associated with the brain-gut axis.

Purpose Ex vivo lung perfusion (EVLP) is an established technology to assess marginal donor lungs. EVLP application has increased the use of donor lungs and is being further developed as a platform for the repair of lung injury. Steen solution is one of the most widely used EVLP perfusate solutions. Currently, clinical EVLP is generally run for 4-6 h, and most of porcine lung EVLP studies were limited to around 12 h. We hypothesize that adding nutrients to Steen solution may improve cellular function for better performance. Methods Human pulmonary microvascular endothelial cells (HPMEC) and human lung epithelial cells (BEAS-2B) were used to determine the effects of Steen solution components on basic cellular function. Cell confluence, apoptosis and migration were visualized and quantified by IncuCyte® Live Cell Analysis System in real-time. A simulated EVLP cell culture model was established by replacing regular culture medium with cold LPD preservation solution and followed by “perfusion” with normothermic Steen solution or its modifications. Cell apoptosis was quantified. Porcine lung EVLP was performed using Steen solution with added nutrients. Results Cells exposed to Steen solution exhibited reduced cell confluence, increased apoptosis and suppressed cell migration compared to DMEM or DMEM+10% FBS. In an examination of adding various nutrients to Steen solution on cell function, L-alanyl-L-glutamine (gln) significantly inhibited cell apoptosis and improved cell migration (Fig A). In the simulated EVLP cell culture setting, adding gln to Steen solution significantly reduced apoptosis. In pig lung EVLP, the addition of gln to Steen solution significantly extend the period of stable lung function (Fig B). Conclusion Glutamine is the most abundant free amino acid in the body with multiple cytoprotective functions. Adding L-alanyl-L-glutamine to EVLP perfusate may improve the stability and function of lungs being evaluated and treated with EVLP.

Jinze Reservoir is the main drinking water source for southwest Shanghai. The water quality in the upstream Taipu River and reservoir have an important impact on the performance of the reservoir and the quality of the water supply. Conventional water parameters were analyzed using principal component analysis (PCA) to evaluate seasonal variations in water quality in the upper reaches of Jinze Reservoir. The parallel factor (PARAFAC) was also used to analyze water-soluble dissolved organic matter (FDOM) in the upstream waters and reservoir area. Furthermore, the relationships between water fluorescence intensity and conventional water quality indicators were analyzed using correlation analysis. The results show that water quality in the upper reaches of Jinze Reservoir is worst in the winter, and water quality gradually deteriorates in the outer sections of the reservoir due to external pollution inputs. Ecological purification within the reservoir area functions to improve water quality. The water body FDOM content is mainly composed of proteinoids and humus. Protein-like substances are the main components of this material, and each component exhibits clear spatial and temporal distribution characteristics. External pollution is the main factor affecting the total fluorescence intensity of the FDOM outside of the reservoir. In the reservoir, the ecological environment affects the total fluorescence intensity of FDOM. The total fluorescence intensity of FDOM showed an increasing downstream spatial trend, which was significantly correlated with NH

In order to obtain a better fermentation parameter for the production of recombinant

BackgroundAlthough the action mechanism of antineoplastic agents is different, oxaliplatin, paclitaxel, or bortezomib as first-line antineoplastic drugs can induce painful neuropathy. In rodents, mechanical allodynia is a common phenotype of painful neuropathy for 3 chemotherapeutics. However, whether there is a common molecular involved in the different chemotherapeutics-induced painful peripheral neuropathy remains unclear.MethodsMechanical allodynia was tested by von Frey hairs following i.p. injection of vehicle, oxaliplatin, paclitaxel, or bortezomib in Sprague-Dawley rats. Reduced representation bisulfite sequencing and methylated DNA immunoprecipitation were used to detect the change of DNA methylation. Western blot, quantitative polymerase chain reaction, chromatin immunoprecipitation, and immunohistochemistry were employed to explore the molecular mechanisms.ResultsIn 3 chemotherapeutic models, oxaliplatin, paclitaxel, or bortezomib accordantly upregulated the expression of transient receptor potential cation channel, subfamily C6 (TRPC6) mRNA and protein without affecting the DNA methylation level of TRPC6 gene in DRG. Inhibition of TRPC6 by using TRPC6 siRNA (i.t., 10 consecutive days) relieved mechanical allodynia significantly following application of chemotherapeutics. Furthermore, the downregulated recruitment of DNA methyltransferase 3 beta (DNMT3b) at paired box protein 6 (PAX6) gene led to the hypomethylation of PAX6 gene and increased PAX6 expression. Finally, the increased PAX6 via binding to the TPRC6 promoter contributes to the TRPC6 increase and mechanical allodynia following chemotherapeutics treatment.ConclusionsThe TRPC6 upregulation through DNMT3b-mediated PAX6 gene hypomethylation participated in mechanical allodynia following application of different chemotherapeutic drugs.

Purpose Ex vivo lung perfusion (EVLP) has emerged as an important platform to assess marginal donor lungs prior to transplantation. Derived from the outer membrane of gram-negative bacteria, lipopolysaccharide (LPS) can stimulate strong immune responses. Studies have shown that donor lungs infected with bacteria may have worse outcomes, suggesting bacteria-induced donor lung injury. However, no large-scale studies have looked at the relationship between LPS levels and donor lung performance on EVLP. Methods LPS levels were measured after one and four hours of perfusion in n=100 clinical EVLP cases. To assess the effects of LPS on lung inflammation, IL-6, 8, 10, and 1β, sTNF-R1, sTREM-1 and ET-1 levels were measured with an ELISA-based platform. Biochemical and physiological assessments were compared to LPS levels. Finally, LPS levels and recipient intensive care unit length of stay after transplantation were investigated. Results There was a significant increase from 0.14 [0.09-0.21] to 0.23 [0.13-0.38] EU/mL (p Conclusion This study demonstrates the trends and importance of circulating LPS levels during EVLP assessment of human donor lung injury. We show that high LPS levels and increasing levels over time were associated with poor donor lung performance in EVLP. These results support the need for therapeutic strategies to target LPS during EVLP, which may improve lung function and rescue more donor lungs for transplantation.

Purpose During lung transplantation, donor biopsies are collected for histologic and biomarker assessments to understand lung injury. Samples are often taken from a single location and it is unknown whether this is truly representative of the injury across the organ. The objective of this study was to investigate the uniformity of cytokine mRNA expression in tissue samples collected from different locations in donor lungs. Methods Eight unused donor lungs were inflated, flash-frozen in liquid nitrogen, and partitioned into 10 to 15 slices from apex to base. Five 1 cm3 biopsies were taken from each lung; three were consistently taken from the third, sixth, and ninth slices, while two were taken from the lingula and a noted potential injury site. Three pieces of each biopsy were used to measure mRNA expression of IL-6, IL-8, IL-10, and IL-1β as markers of lung inflammation by PCR in a total of n=120 samples from the eight lungs. Intra-biopsy and intra-lung variations were analyzed with equal-variance tests, coefficients of variance, one-way ANOVA, and pairwise mean comparisons. Results Across all donors, the intra-biopsy equal-variance F-values ranged from 0.19-0.95 with means of 0.33, 0.57, 0.62, and 0.58 for IL-1β, IL-6, IL-8, and IL-10. The mean biopsy coefficient of variance was 25.9% across all donors. The mean comparisons of biopsies in each donor showed that the three consistent slices were identical in donors rejected for logistics and thrombotic emboli. Both the lingula and focal injury biopsies demonstrated larger differences in cytokine expression from the rest of the donor lung. Conclusion While intra-biopsy variances were present, they were small and consistent across multiple biopsies in donors, indicating the reliability of repeated analysis of banked specimens. Lungs rejected for non-graft related reasons demonstrated more consistent gene expression. The lingula is not a representative site as it often differed from other biopsies in all donors. We conclude that a donor lung biopsy taken from an area other than the lingula reflects the overall condition of the lung, unless there is obvious localized injury (e.g. infection) which has a unique profile. These findings will reassure the scientific community that a biopsy of the donor lung is indeed representative of the whole lung, and will help to guide diagnostic and therapeutic interventions in donor lungs for transplantation.

Although the anxiolytic-like effects of Xiaoyaosan, a Chinese herbal formula, have been described in many previous studies, its underlying mechanism remains undefined. The cytokine tumour necrosis factor-α (TNF-α) and its closely associated janus kinase 2 (JAK2)-signal transducer and activator of transcription (STAT3) signalling pathway regulate the neuro-inflammatory response in the brain, thus participating in the development of anxiety. Our purpose was to investigate whether the anxiolytic-like effects of Xiaoyaosan are related to the TNF-α/JAK2-STAT3 pathway in the hippocampus. We examined the effects of Xiaoyaosan on behaviours exhibited in the elevated plus maze test, open field test and novelty-suppressed feeding test as well as hippocampal neuron damage and changes in the TNF-α/JAK2-STAT3 pathway in a rat model of chronic immobilization stress (CIS)-induced anxiety. Xiaoyaosan exerts anxiolytic-like effects on CIS-induced anxiety, with a significant alleviation of anxiety-like behaviours, an attenuation of hippocampal neuron damage, and a reversal of the activation of the TNF-α/JAK2-STAT3 pathway in the hippocampus that are similar to the effects of the JAK2 antagonist AG490. However, Xiaoyaosan and AG490 failed to effectively regulate apoptosis-related factors, including Bax and Caspase-3. These results suggest that Xiaoyaosan attenuates stress-induced anxiety behaviours by down-regulating the TNF-α/JAK2-STAT3 pathway in the rat hippocampus.

// Serisha Moodley 1,2 , Mathieu Derouet 2 , Xiao Hui Bai 2 , Feng Xu 3 , Andras Kapus 1,4 , Burton B. Yang 1,5 and Mingyao Liu 1,2 1 Institute of Medical Science, Faculty of Medicine, University of Toronto, Toronto, Canada 2 Latner Thoracic Surgery Research Laboratories, Toronto General Research Institute, University Health Network, Toronto, Canada 3 Advanced Optical Microscopy Facility, UHN, Toronto, Canada 4 Keenan Research Centre for Biomedical Science, St. Michael’s Hospital, Toronto, Canada 5 Sunnybrook Research Institute, Sunnybrook Health Sciences Centre, Toronto, Canada Correspondence to: Mingyao Liu, email: // Keywords : actin filament associate protein 1-like 2, SH3PXD2A, lamellipodia, podosomes, lung repair, Pathology Section Received : August 26, 2016 Accepted : November 02, 2016 Published : November 09, 2016 Abstract Repair of airway epithelium after injury requires migration of neighboring epithelial cells to injured areas. However, the molecular mechanisms regulating airway epithelial cell migration is not well defined. We have previously shown that XB130, a scaffold protein, is required for airway epithelial repair and regeneration in vivo , and interaction between XB130 and another scaffold protein, Tks5, regulates cell proliferation and survival in human bronchial epithelial cells. The objective of the present study was to determine the role of XB130 and Tks5 interaction in airway epithelial cell migration. Interestingly, we found that XB130 only promotes lateral cell migration, whereas, Tks5 promotes cell migration/invasion via proteolysis of extracellular matrix. Upon stimulation with EGF, PKC activator phorbol 12, 13-dibutyrate or a nicotinic acetylcholine receptor ligand, XB130 and Tks5 translocated to the cell membrane in a stimulus-dependent manner. The translocation and distribution of XB130 is similar to lamellipodial marker, WAVE2; whereas Tks5 is similar to podosome marker, N-WASP. Over-expression of XB130 or Tks5 alone enhances cell migration, whereas co-expression of both XB130 and Tks5 inhibits cell migration processes and signaling. Furthermore, XB130 interacts with Rac1 whereas Tks5 interacts with Cdc42 to promote Rho GTPase activity. Our results suggest that dissociation between XB130 and Tks5 may facilitate lateral cell migration via XB130/Rac1, and vertical cell migration via Tks5/Cdc42. These molecular mechanisms will help our understanding of airway epithelial repair and regeneration.

Clinical application of hydrophobic therapeutics is restricted by lack of an efficient vehicle which permits their solubility in aqueous environments. We have previously developed a novel formulation strategy to deliver a hydrophobic Src inhibitor, PP2, involving combinations of one self-assembling peptide (SAP) and one of 4 selected amino acids (AAs). The present study aims to develop a generalized drug delivery platform for intravenous application of hydrophobic drugs by combining self-assembling peptide, amino acid and low concentration of co-solvent. A multi-step screening pipeline is established which includes assessment of drug solubility and physicochemical characteristics, as well as functional efficacy and safety in vitro and in vivo. Using PP2 as an exemplary hydrophobic compound, 480 different combinations of 6 SAPs, 20 naturally existing AAs at 2 concentrations, and 2 co-solvents were evaluated. Among the combinations, 60 formulae dissolved PP2; 10 of which significantly reduced thrombin-induced IL-8 production, a sign of inflammatory response, in normal human lung epithelial BEAS2B cells. These formulations did not show cytotoxicity alone, but 2 reduced cell viability with presence of thrombin. We then performed a double-blinded test in a rat model of pulmonary ischemia-reperfusion. PP2 formulated with EAK16-I peptide plus methionine and 2% ethanol were administrated intravenously, significantly reducing severity of lung injury. The SAP-AA formulation strategy was also successfully applied to other hydrophobic compounds, suggesting this strategy could be applicable to other hydrophobics for a variety of clinical applications.

Objective: To investigate the influencing factors of blood pressure variability (BPV) in patients with maintenance hemodialysis (MHD), so as to improve the patients’ prognosis.Methods: The clinical data of 107 MHD patients were retrospectively analyzed. According to intradialytic systolic pressure (SBP)-BPV, the patients were assigned into high SBP-BPV group (n=52) and low SBP-BPV group (n=55). According to intradialytic diastolic pressure (DBP)-BPV, they were divided into high DBP-BPV group (n=49) and low DBP-BPV group (n=58). The basic characteristics of patients in high and low SBP-BPV groups and DBP-BPV groups were compared, and the influencing factors of both SBP-BPV and DBP-BPV were also analyzed.Results: The differences were statistically significant between high and low SBP-BPV groups by comparison to the age, dry weight, body mass index (BMI), dialysis age, interdialysis weight gain (IDWG) rate, pre-dialysis SBP, albumin (ALB), hemoglobin (Hb), total cholesterol (TC) and calcium-phosphorus product (P

IntroductionTransplantation of lungs from donation after circulatory death (DCD) in addition to donation after brain death (DBD) became routine worldwide to address the global organ shortage. The development ofex vivolung perfusion (EVLP) for donor lung assessment and repair contributed to the increased use of DCD lungs. We hypothesise that a better understanding of the differences between lungs from DBD and DCD donors, and between EVLP and directly transplanted (non-EVLP) lungs, will lead to the discovery of the injury-specific targets for donor lung repair and reconditioning.MethodsTissue biopsies from human DBD (n=177) and DCD (n=65) donor lungs, assessed with or without EVLP, were collected at the end of cold ischaemic time. All samples were processed with microarray assays. Gene expression, network and pathway analyses were performed using R, Ingenuity Pathway Analysis and STRING. Results were validated with protein assays, multiple logistic regression and 10-fold cross-validation.ResultsOur analyses showed that lungs from DBD donors have upregulation of inflammatory cytokines and pathways. In contrast, DCD lungs display a transcriptome signature of pathways associated with cell death, apoptosis and necrosis. Network centrality revealed specific drug targets to rehabilitate DBD lungs. Moreover, in DBD lungs, tumour necrosis factor receptor-1/2 signalling pathways and macrophage migration inhibitory factor-associated pathways were activated in the EVLP group.A panel of genes that differentiate the EVLP from the non-EVLP group in DBD lungs was identified.ConclusionThe examination of gene expression profiling indicates that DBD and DCD lungs have distinguishable biological transcriptome signatures.