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Licochalcone (LicA) is a flavonoid commonly derived from the licorice plant that is reported to have a variety of pharmacological activities. However, few studies have focused on its anti-allergic properties. IgE-mediated passive and systemic anaphylaxis mice models were used to assess the in vivo anti-allergic effect of LicA and its underlying mechanism, while degranulation, cytokines, and chemokines released from laboratory of allergic disease (LAD2) cells were used to assess its in vitro anti-allergic effect. We used western blot analysis to explore the downstream signaling pathway of its anti-allergic effect. We found that in the mouse model, LicA attenuated IgE-mediated paw inflammation, recovered the allergy-induced drop in body temperature, and reduced the concentrations of tumor necrosis factor-alpha and monocyte chemo-attractant protein-1 in mouse serum in a dose-dependent manner. LicA inhibited the allergic reaction via inhibition of IgE-mediated LAD2 cell activation through the PLC/ERK/STAT3 pathway.

Menaquinone-7 (MK-7) possesses wide health and medical value, and the market demand for MK-7 has increased. Metabolic engineering for MK-7 production in Escherichia coli still remains challenging due to the characteristics of the competing quinone synthesis, and cells mainly synthesized menaquinones under anaerobic conditions. To increase the production of MK-7 in engineered E. coli strains under aerobic conditions, we divided the whole MK-7 biosynthetic pathway into three modules (MVA pathway, DHNA pathway, and MK-7 pathway) and systematically optimized each of them. First, by screening and enhancing Idi expression, the amounts of MK-7/DMK-7 increased significantly. Then, in the MK-7 pathway, by combinatorial overexpression of endogenous MenA and exogenous UbiE, and fine-tuning the expression of HepPPS, MenA, and UbiE, 70 μM MK-7 was achieved. Third, the DHNA synthetic pathway was enhanced, and 157 μM MK-7 was achieved. By the combinational metabolic engineering strategies and membrane engineering, an efficient metabolic engineered E. coli strain for MK-7 synthesis was developed, and 200 μM (129 mg/L) MK-7 was obtained in shake flask experiment, representing a 306-fold increase compared to the starting strain. In the scale-up fermentation, 2074 μM (1350 mg/L) MK-7 was achieved after 52 h fermentation with a productivity of 26 mg/L/h. This is the highest titer of MK-7 ever reported. This study offers an alternative method for MK-7 production from biorenewable feedstock (glucose) by engineered E. coli. The high titer of our process should make it a promising cost-effective resource for MK-7.

HighlightsUsing an elevated platform with a manure catcher reduced litter moisture content and ammonia concentration.A robotic vehicle encouraged bird movement and use of the elevated platform.Using the elevated platform and robotic vehicle jointly improved broiler paw quality and plumage cleanliness.Abstract. Elevated platform (EP) and robotic vehicle (RV) are two emerging systems aiming to improve environment enrichment and bird activity in broiler housing systems. However, the impacts of these systems on broiler production, welfare, and housing environment have not been evaluated. The objective of this study was to evaluate the effects of using EP and RV individually or jointly on broiler feed conversion ratio (FCR), litter moisture content (LMC), ammonia (NH3) concentration, gait score (GS), paw quality (PQ), plumage cleanliness (PC), and bird activity index (AI). Broilers were reared for eight-week production cycles. Four experimental rooms (54 birds room-1) were randomly assigned four treatments: EP only, RV only, EP and RV (EP+RV), and neither EP nor RV (Ctrl). Broiler GS, PQ, and PC were assessed following welfare protocols. Bird AI was determined through image processing. The experiment was repeated three times. The results showed that overall FCR values were 1.806 for EP, 1.804 for RV, 1.797 for EP+RV, and 1.791 for Ctrl. The normalized LMC values were 23.2% ±4.1% for EP, 32.8% ±4.1% for RV, 23.4% ±2.5% for EP+RV, and 35.7% ±7.0% for Ctrl over the eight-week production cycles. NH3 concentrations were 40% lower in the rooms with EP than in the rooms without EP at the end of the production cycle. Broilers had better PQ in the rooms with EP than in the rooms without EP. Broiler PC seemed better in the rooms with RV compared to those without RV. Operation of RV increased bird AI; however, the benefits in activity encouragement diminished as the broilers grew. The number of broilers on the EP was higher in the EP+RV rooms than in the EP rooms. It is concluded that using EP and RV together may improve broiler welfare and activity without compromising their production performance. Keywords: Behavior, Broiler, Elevated platform, Robotic vehicle, Welfare.

Hepatocellular carcinoma (HCC), the most common type of primary liver cancer, is mainly due to genetic changes in hepatocytes. However, molecular expression in hepatocytes during hepatocarcinogenesis has not been characterized. In this study, using an inducible kras transgenic zebrafish models for HCC, transcriptomic profiles of oncogenic hepatocytes from larvae, male and female adult fish following a brief induction of oncogenic kras were investigated. We found that oncogenic hepatocytes from all the three sources possess most of the cancer hallmarks at molecular level, including Sustaining proliferative signaling, Evading growth suppressors, Resisting cell death, Avoiding immune destruction, Inflammation, Reprogramming of energy metabolism, Angiogenesis, and Activating invasion and metastasis, suggesting the malignant transformation at molecular level could occur at the early stage of hepatocarcinogensis and can be captured in hepatocytes. However, each group of oncogenic hepatocytes also had their own characteristics. Larval oncogenic hepatocytes have cancer stem cell features. Female oncogenic hepatocytes showed resemblance to a mild human HCC subtype while male oncogenic hepatocytes resembled a severe HCC subtype, consistent with the observed sex disparity of HCC in both zebrafish and human. Finally, the two adult groups were more similar to each other than to the larval group, indicating an overwhelming effect of development over the gender.

Gadopentetate meglumine (Gd-DTPA) is a commonly used magnetic resonance imaging (MRI) enhancer in the clinic for improving the clarity of MRI. Reports have shown that severe anaphylactoid reactions can occur after intravenous injections, whereas the underlying mechanisms remain undefined. In this study, the effects of Gd-DTPA in causing allergic like reactions and mast cells (MCs) activation were investigated both in vitro and in vivo. Gd-DTPA induced a severe hand paw swelling and body temperature decrease in murine topical cutaneous allergy model, and murine systemic allergy model. Meanwhile, serum IgE was not significantly changed. Histamine, tryptase, and cytokines release in mice serum and LAD2 cells supernatant were increased significantly both in mice serum and LAD2 supernatant after treated with Gd-DTPA. Subsequently, the changes in mRNA levels after Gd-DTPA activated MCs were analyzed by RNAseq and found to be related to inflammation signaling pathways. The study provides a demonstration for the adverse reaction mechanism of Gd-DTPA and its safe use in clinic.

Menaquinone-7 (MK-7) possesses wide health and medical value, and the market demand for MK-7 has increased. Metabolic engineering for MK-7 production in

Environmental pollutants may cause adverse effects on the immune system of aquatic organisms. However, the cellular effects of pollutants on fish immune system are largely unknown. Here, we exploited the transgenic zebrafish Tg(lysC:DsRed2) larva as a preliminary screening system to evaluate the potential inflammatory effects of environmental pollutants. Tg(lysC:DsRED2) larvae aged 7-day-postfertilization (7 dpf) were treated with selected environmental chemicals for 24 h (24 h) and the number of neutrophils were quantified using both image analysis and fluorescence activated cell sorting (FACS). We found that the numbers of neutrophils in the Tg(lysC:DsRED2) larvae were significantly increased by most of the organic chemicals tested, including E2 (17β-estradiol), BPA (Bisphenol-A), NDEA (N-nitrosodiethylamine), 4-NP (4-Nitrophenol) and Lindane (γ-hexachlorocyclohexane). Neutrophil numbers were also increased by all the metals tested (Na2HAsO4· 7H2O, Pb(NO3)2, HgCl2, CdCl2, CuSO4·5H2O, ZnSO4, and K2Cr2O7). The only exception was TCDD (2,3,7,8-tetrachlorodibenzo-p-dioxin), which significantly reduced the number of neutrophils after exposure. Additionally, the transcription of genes (lyz, mpo, tnfα and il8) related to fish immune system were significantly modulated upon exposure to some of the selected chemicals such as E2, TCDD, Cu and Cd. This study revealed that representatives of major categories of environmental pollutants could cause an acute inflammatory response in zebrafish larvae as shown by alterations in the neutrophils, which may imply a common immunotoxicity mechanism for most environmental pollutants. This study has also demonstrated that Tg(lyz:DsRed2) transgenic zebrafish is an excellent tool for screening environmental chemicals with potential inflammatory effects through FACS-facilitated neutrophil counting.

Introduction Many cancers involve the participation of rare cell populations that may only be found in a subset of patients. Single-cell RNA sequencing (scRNA-seq) can identify distinct cell populations across multiple samples with batch normalization used to reduce processing-based effects between samples. However, aggressive normalization obscures rare cell populations, which may be erroneously grouped with other cell types. There is a need for conservative batch normalization that maintains the biological signal necessary to detect rare cell populations. MapBatch We designed a batch normalization tool, MapBatch, based on two principles: an autoencoder trained with a single sample learns the underlying gene expression structure of cell types without batch effect; and an ensemble model combines multiple autoencoders, allowing the use of multiple samples for training. Each autoencoder is trained on one sample, learning a projection into the biological space S representing the real expression differences between cells in that sample (Figure 1a, middle). When other samples are projected into S, the projection reduces expression differences orthogonal to S, while preserving differences along S. The reverse projection transforms the data back into gene space at the autoencoder's output, sans expression differences orthogonal to S (Figure 1a, right). Since batch-based technical differences are not represented in S, this transformation selectively removes batch effect between samples, while preserving biological signal. The autoencoder output thus represents normalized expression data, conditioned on the training sample. To incorporate multiple samples into training, MapBatch uses an ensemble of autoencoders, each trained with a single sample (Figure 1b). We train with a minimal number of samples necessary to cover the different cell populations in the dataset. We implement regularization using dropout and noise layers, and an a priori feature extraction layer using KEGG gene modules. The autoencoders' outputs are concatenated for downstream analysis. For visualization and clustering, we use the top principal components of the concatenated outputs. For differential expression (DE), we perform DE on each of the gene matrices output by each model, then take the result with the lowest P-value. To test MapBatch, we generated a synthetic dataset based on 7 batches of publicly available PBMC data. For each batch we simulated rare cell populations by selecting one of three cell types to perturb by up and down-regulating 40 genes in 0.5%-2% of the cells (Figure 1c). We simulated additional batch effect by scaling each gene in each batch with a scaling factor. Upon visualization and clustering, cells grouped largely by batch (Figure 1d). After batch normalization, cells grouped by cell type rather than batch, and all three perturbed cell populations were successfully delineated (Figure 1e). DE between each perturbed population and its mother cells accurately retrieved the perturbed genes, showing that normalization maintained real expression differences (Figure 1e). In contrast, three methods tested Seurat (Stuart et al., 2019), Harmony (Korsunsky et al., 2019), and Liger (Welch et al., 2019) could only derive a subset of the perturbed populations (Figures 1f-h). MapBatch identifies rare populations in multiple myeloma (MM) We used MapBatch to process bone marrow scRNA-seq data from 14 MM samples and 2 healthy controls. After batch normalization, unsupervised clustering identified 20 clusters, which we annotated using MapCell (Koh & Hoon, 2019) (Figures 2a, 2b). We identified 3 small clusters of cells that could not be reliably annotated, comprising less than 1% of total cells and found in only a subset of patients (Figures 2c, 2d). As validation, we observed that these cells were present in distinct clusters in individual samples using their uncorrected expression data, providing evidence that these clusters were not driven by batch effect nor MapBatch (Figure 2e). Conclusion Batch normalization of scRNA-seq data involves a trade-off between minimizing batch effect and maximizing the remaining biological signal. While most methods lean towards the former, MapBatch maintains more biological signal for downstream analysis, enabling the discovery of previously difficult to find cell populations. Figure 1 Figure 1. Disclosures Xu: Proteona Pte Ltd: Current Employment. Scolnick: Proteona Pte Ltd: Current holder of individual stocks in a privately-held company. Huo: Proteona Pte Ltd: Ended employment in the past 24 months. Lovci: Proteona Pte Ltd: Current Employment. Chng: Amgen: Honoraria, Research Funding; Abbvie: Honoraria; Janssen: Honoraria, Research Funding; Novartis: Honoraria; Celgene: Honoraria, Research Funding.

Background Multiple Myeloma (MM) is an incurable plasma cell (PC) malignancy and high risk MM remains an unmet clinical need. Translocation 4;14 occurs in 15% of MM and is associated with an adverse prognosis. A deeper understanding of the biology and immune micro-environment of t(4;14) MM is necessary for the development of effective targeted therapies. Single Cell multi-omics provides a new tool for phenotypic characterization of MM. Here we used Proteona's ESCAPE™ single cell multi-omics platform to study a cohort of patients with t(4;14) MM. Methods Diagnostic bone marrow (BM) samples from 13 patients with t(4;14) MM (one of whom had samples at diagnosis and relapse) were analysed using the ESCAPE™ platform from Proteona which simultaneously measures gene and cell surface protein expression of 65 proteins in single cells. Cryopreserved BM samples were stained with antibodies and subsequently sorted on CD138 expression. The CD138 positive and negative fractions were recombined at a 1:1 ratio for analysis using the 10x Genomics 3' RNAseq kit. Resulting data were analyzed with Proteona's MapSuite™ single cell analytics platform. In particular, Mapcell was used to annotate the cells and MapBatch was used for batch normalization in order to preserve rare cell populations. Results Patients had a median age of 63 years and received novel agent-based induction. Median progression free and overall survival (PFS and OS) were 22 and 34 months respectively. We first analyzed serial BM samples from an individual patient that were taken at diagnosis and relapse following bortezomib based treatment. The PCs in this patient showed variations in gene expression between diagnosis and relapse (Fig 1A), including the reduction of HIST1H2BG expression, which has previously been correlated with resistance to bortezomib. Subsequent analysis of the immune cells identified a shift in the ratio of T cells to CD14 monocytes from 5.7 at diagnosis to 0.6 at relapse suggesting a major change in the BM immune micro-environment in response to therapy. Next, we analyzed the malignant PCs of the diagnostic samples. As expected, MMSET (NSD2) was overexpressed in all PCs compared to normal PCs, while FGFR3 expression could be categorized into no expression of FGFR3, low expression (80% of cells expressing FGFR3) (Fig 1B). No gene or protein expression patterns within the PCs were identified that correlated with PFS or OS in this cohort. Finally, we analyzed the immune micro-environment in the diagnostic samples (Fig 1C). While there was no overall discernable pattern of cell types present, one cluster of cells, annotated as 'unknown' cell type, suggested a small population of cells that had not been previously annotated in published single cell RNA-seq data. The cells were CD45+ and CD138 - both at the protein and RNA level, suggesting they are not plasma cells. We tested if the number of the 'unknown' cells in each sample correlated with PFS, but there was no significant correlation. We then used these cells to derive a gene signature profile which was expressed in most of the cells in the 'unknown' cluster as well as a minor fraction of cells in other clusters including some PCs. The number of cells expressing the gene signature negatively correlated with PFS, with samples containing more cells expressing the signature having a lower PFS than samples with fewer signature positive cells (Fig 2). The correlation remained significant whether we included PCs in the analysis or not, but was not significant amongst only the PC population, suggesting that the cells responsible for the correlation are from the immune micro-environment. Conclusions We present the first application of single cell multi-omic immune profiling in high-risk MM and demonstrate that t(4;14) is a phenotypically heterogenous disease. While no consistent gene or protein expression patterns were identified within the malignant cell population, we did identify gene expression changes in a relapsed patient sample that may reflect key alterations in the PCs responsible for therapy resistance. In addition, we identified a gene signature expressed in a rare population of non-plasma cells that significantly correlated with PFS in this patient cohort. These data highlight the potential of single cell multi-omic analysis to identify immune micro-environmental signatures that correlate with response to therapy in t(4;14) MM. Figure 1 Figure 1. Disclosures Scolnick: Proteona Pte Ltd: Current holder of individual stocks in a privately-held company. Huo: Proteona Pte Ltd: Ended employment in the past 24 months. Xu: Proteona Pte Ltd: Current Employment. Chng: Amgen: Honoraria, Research Funding; Janssen: Honoraria, Research Funding; Celgene: Honoraria, Research Funding; Novartis: Honoraria; Abbvie: Honoraria.

Context Multiple myeloma (MM) is an incurable plasma cell (PC) malignancy and high risk (HR) MM remains an unmet clinical need. Translocation 4;14 occurs in 15% of MM and is associated with an adverse prognosis. A deeper understanding of the biology and immune micro-environment of t(4;14) MM is necessary for the development of effective targeted therapies. Objective Here we utilized Proteona’s ESCAPE™ single cell multi-omics platform to study a cohort of patients with t(4;14) MM with the goal of better understanding the biology underlying this high risk patient cohort. Design Diagnostic bone marrow (BM) samples from 13 patients with t(4;14) MM were analysed using the ESCAPE platform from Proteona which simultaneously measures gene and cell surface protein expression in single cells. Resulting data were analyzed with MapSuite tools from Proteona and Seurat. Results The patients had a median age of 63 years. All received novel agent based induction. Median progression free and overall survival (PFS and OS) were 22 and 34 months respectively. MMSET was overexpressed in all PCs while FGFR3 expression could be categorized into low expression ( 80% of cells expressing FGFR3). Variation in the immune microenvironment of the BM was seen across all patient samples with no correlation between cell types present in the BM and PFS or OS. Plasma cells from the combined samples formed clusters that could be differentiated by gene expression with each cluster having a unique combination of cells of varying PFS. Gene and protein expression differences were seen in PCs when samples were clustered by PFS suggesting a role for these molecules in disease progression. Finally, we identified gene expression changes in one patient between a diagnostic sample and relapse disease. Conclusions We present the first application of single cell multi-omics immune profiling in high-risk MM. Our results suggest that t(4;14) MM is a molecularly heterogeneous disease. That heterogeneity, coupled with our small sample size may explain the lack of correlation between gene or protein expression with clinical outcomes. Single cell analysis of larger cohorts is required to build on our findings.

e20013 Background: Multiple Myeloma (MM) is an incurable plasma cell (PC) malignancy and high risk (HR) MM remains an unmet clinical need. Translocation 4;14 occurs in 15% of MM and is associated with an adverse prognosis. A deeper understanding of the biology and immune micro-environment of t(4;14) MM is necessary for the development of effective targeted therapies. Single Cell multi-omics provides a new tool for phenotypic characterization of MM. Here we used Proteona’s ESCAPE™ single cell multi-omics platform to study a cohort of patients with t(4;14) MM. Methods: Diagnostic bone marrow (BM) samples from 14 patients with t(4;14) MM were analysed using the ESCAPE platform from Proteona which simultaneously measures gene and cell surface protein expression in single cells. Cryopreserved BM samples were stained with 65 DNA barcoded antibodies and subsequently sorted on CD138 expression. The CD138 positive and negative fractions were recombined at a known ratio for analysis using the 10x Genomics 3’ RNAseq kit. Resulting data were analyzed with Seurat and MapCell. Results: The patients had a median age of 63 years. All received novel agent based induction. Median progression free and overall survival (PFS and OS) were 22 and 34 months respectively. MMSET was overexpressed in all PCs while FGFR3 expression could be categorized into zero cells expressing FGFR3, low expression (< 10% of cells expressing FGFR3) or high expression (> 80% of cells expressing FGFR3). We also found heterogeneity in the expression of cancer testis antigens (CTA) such as FA133A and CTAG2 between PC clusters across samples. Variation in the immune microenvironment of the BM was seen across all patient samples with no correlation between cell types and PFS or OS. However, an analysis of BM samples at diagnosis and relapse in one patient showed a shift in the ratio of T cells to CD14 monocytes with a ratio of 5.7 at diagnosis compared to 0.6 at relapse. Further analysis of PCs in this patient found 8 PC populations, each containing variable numbers of cells from both the diagnostic and relapse samples. This suggests that all populations present at relapse were also present at diagnosis, although at variable proportions. Increased expression of RCAN3 (associated with cereblon depletion) was detected at relapse. Conclusions: We present the first application of single cell multi-omics immune profiling in high risk MM. The heterogeneity in expression of CTA has implications for the application of immunotherapies, while the upregulation of RCAN3 may explain failure of immunomodulatory therapy. Our small sample size may explain the lack of correlation between gene or protein expression with clinical outcomes. We propose that t(4;14) MM is a genomically and immunologically heterogeneous disease. Single cell analysis of larger cohorts is required to build on our findings.

Hepatocellular carcinoma (HCC) is one of the most severe cancer types and many genetic and environmental factors contribute to the development of HCC. Androgen receptor (AR) signaling is increasingly recognized as one of the important factors associated with HCC. Previously, we have developed an inducible HCC model in kras transgenic zebrafish. In the present study, to investigate the role of AR in liver tumor development, we specifically knocked out ar gene in the liver of zebrafish via the CRISPR/Cas9 system and the knockout zebrafish was named L-ARKO for liver-specific ar knockout. We observed that liver-specific knockout of ar attenuated liver tumor development in kras transgenic zebrafish at the early stage (one week of tumor induction). However, at the late stage (two weeks of tumor induction), essentially all kras transgenic fish continue to develop HCC irrespective of the absence or presence of ar gene, indicating an overwhelming role of the driver oncogene kras over ar knockout. Consistently, cell proliferation was reduced at the early stage, but not the late stage, of liver tumor induction in the kras/L-ARKO fish, indicating that the attenuant effect of ar knockout was at least in part via cell proliferation. Furthermore, androgen treatment showed acceleration of HCC progression in kras fish but not in kras/L-ARKO fish, further indicating the abolishment of ar signalling. Therefore, we have established a tissue-specific ar knockout zebrafish and it should be a valuable tool to investigate AR signalling in the liver in future.

We have previously demonstrated the pro-tumoral role of neutrophils using a kras-induced zebrafish hepatocarcinogenesis model. To further illustrate the molecular basis of the pro-tumoral role, Tumor-associated neutrophils (TANs) were isolated by fluorescence-activated cell sorting (FACS) and transcriptomic analyses were carried out by RNA-Seq. Differentially expressed gene profiles of TANs from larvae, male and female livers indicate great variations during liver tumorigenesis, but the common responsive canonical pathways included an immune pathway (Acute Phase Response Signaling), a liver metabolism-related pathway (LXR/RXR Activation) and Thrombin Signaling. Consistent with the pro-tumoral role of TANs, gene module analysis identified a consistent down-regulation of Cytotoxicity module, which may allow continued proliferation of malignant cells. Gene Set Enrichment Analysis indicated up-regulation of several genes promoting angiogenesis. Consistent with this, we found decreased density of blood vessels accompanied with decreased oncogenic liver sizes in neutrophil-depleted larvae. Collectively, our study has indicated some molecular mechanisms of the pro-tumoral roles of TANs in hepatocarcinogenesis, including weakened immune clearance against tumor cells and enhanced function in angiogenesis.

Hepatocellular carcinoma (HCC), the most common type of primary liver cancer, is mainly due to genetic changes in hepatocytes. However, molecular expression in hepatocytes during hepatocarcinogenesis has not been characterized. In this study, using an inducible kras transgenic zebrafish models for HCC, transcriptomic profiles of oncogenic hepatocytes from larvae, male and female adult fish following a brief induction of oncogenic kras were investigated. We found that oncogenic hepatocytes from all the three sources possess most of the cancer hallmarks at molecular level, including Sustaining proliferative signaling, Evading growth suppressors, Resisting cell death, Avoiding immune destruction, Inflammation, Reprogramming of energy metabolism, Angiogenesis, and Activating invasion and metastasis, suggesting the malignant transformation at molecular level could occur at the early stage of hepatocarcinogensis and can be captured in hepatocytes. However, each group of oncogenic hepatocytes also had their own characteristics. Larval oncogenic hepatocytes have cancer stem cell features. Female oncogenic hepatocytes showed resemblance to a mild human HCC subtype while male oncogenic hepatocytes resembled a severe HCC subtype, consistent with the observed sex disparity of HCC in both zebrafish and human. Finally, the two adult groups were more similar to each other than to the larval group, indicating an overwhelming effect of development over the gender.

Conventional infrared spectroscopy is used to investigate the degradation region after electrical treeing in polymers. However, the dimension of the infrared light spot is too large compared with the tree channel, and the position of the light spot is imprecise. Consequently, the micro-infrared spectroscopy is proposed in this paper, which is aim to do chemical analysis for electrical tree area in polyethylene at a micrometer scale. A ramped ac voltage was applied for electrical tree inception in polyethylene, utilizing a wire-plane electrode structure. Slices of samples with electrical trees were then analyzed through micro-infrared spectroscopy. The characteristic group distribution within a selected treeing area was provided through micro-infrared mapping. The experiment results indicated the existence of C=O, C-OH and C-O-C groups, and the content of C-OH and C-O-C groups was more than C=O group. The appearance of C=O group was accompanied with the C-OH group, and the latter absorbance is higher, so it is inferred the C=O group tend to degrade further according to the Norrish type II regime. In addition, not all the electrical tree area had the oxygen containing groups. The material degradation during electrical treeing was discussed with and without oxygen.

Previously we have generated inducible liver tumor models by transgenic expression of an oncogene and robust tumorigenesis can be rapidly induced by activation of the oncogene in both juvenile and adult fish. In the present study, we aimed at chemical intervention of tumorigenesis for understanding molecular pathways of tumorigenesis and for potential development of a chemical screening tool for anti-cancer drug discovery. Thus, we evaluated the roles of several major signaling pathways in krasV12- or Myc-induced liver tumors by using several small molecule inhibitors: SU5402 and SU6668 for VEGF/FGF signaling; IWR1 and cardionogen 1 for Wnt signaling; and cyclopamine and Gant61 for Hedgehog signaling. Inhibition of VEGF/FGF signaling was found to deter both Myc- and krasV12-induced liver tumorigenesis while suppression of Wnt signaling relaxed only Myc- but not krasV12-induced liver tumorigenesis. Inhibiting Hedgehog signaling did not suppress either krasV12 or Myc-induced tumors. The suppression of liver tumorigenesis was accompanied with a decrease of cell proliferation, increase of apoptosis, distorted liver histology. Collectively, our observations suggested the requirement of VEGF/FGF signaling but not the hedgehog signaling in liver tumorigenesis in both transgenic fry. However, Wnt signaling appeared to be required for liver tumorigenesis only in Myc but not krasV12 transgenic zebrafish.

Using our previously established xmrk transgenic zebrafish, hepatocellular carcinoma (HCC) was generated by induced expression of xmrk, which encoded a hyperactive epidermal growth factor receptor (EGFR) homolog, and regressed by suppression of xmrk expression. To investigate molecular changes in liver tumour progression and regression, RNA-Seq was performed for induced HCC and early and late stages of liver tissues during tumour regression. We found that Xmrk-induced zebrafish HCC shared strong molecular characteristics with a human HCC subtype (S2), which shows activated Myc signalling, upregulated phosphor-S6 and epithelial cell adhesion molecule. In the HCC stage, there were enhanced proteasome, antigen processing and presentation, aminosugars metabolisms, p53 and cell cycle pathways. During tumour regression, the transcriptomic profile showed a reversed trend of molecular changes compared with human HCC progression. Interestingly, distinct immune responses in tumour progression and regression were observed, including increased major histocompatibility complex class I (MHCI) at the HCC stage, enriched immune cell trafficking signals and inflammation in early regression and enhanced MHCII in late regression. Both neutrophils and macrophages were enriched during tumour progression and regression; however, the distribution of neutrophils and macrophages in HCC was relatively uniform, whereas both types of immune cells were regionally clustered during tumour regression, especially with dominant blood vessel association of macrophage in late regression, suggesting differential functions of these immune cells in tumour progression and regression. As tumour regression in our model resembles the targeted inhibition of EGFR in cancer therapy, our observations may provide molecular insights into the targeted inhibition and highlight the importance of immune response in tumour regression.

Tris(1,3-dichloro-2-propyl) phosphate (TDCPP) is frequently present in indoor dust and can be detected in human milk. In order to evaluate the effects of TDCPP on vertebrate development, zebrafish embryos/larvae were used as an animal model to examine developmental phenotypes and explore possible mechanisms of toxicity by employing microarrays and iTRAQ labeling quantitative proteomics. The results demonstrated that treatment with TDCPP (3 μM) from 0.75 h postfertilization (hpf) inhibited cell rearrangement at 4 hpf, caused delay in epiboly at 5.7 and 8.5 hpf, and led to abnormal development (e.g., short tail, reduced body size) and lethality between 14 and 45 hpf, which might be related with altered expression of genes regulating embryogenesis. Furthermore, trunk curvature was observed as the main phenotype in 96 hpf zebrafish larvae exposed to 1 or 3 μM TDCPP, possibly by changing somite formation and expression of proteins related to fast muscle and cartilage development. Collectively, our results suggest that exposure to TDCPP causes developmental toxicity in vertebrates and warrant the need for studies to evaluate the potential health risks of TDCPP to developing human embryos/infants/children, due to its frequent presence in indoor dust and potential for human exposure.

The method to manage the remaining life of existing AC underground lines was introduced by CIGRE TB 358. In this paper, the evaluation method was modified and used to assess the condition of 10 kV distribution cables of Futian District in Shenzhen and optimize the maintenance strategy. The technology, economy, and strategy scores were considered for the final scores of the cables. The technical-part scores were mainly determined based on the operational years, operational environment, load conditions, laying and fault history of these cables. Finally, the preventive test and management strategy for these cables in the future were suggested according to the final scores.

This study analyzes the low-frequency dielectric and dynamic mechanical properties of the cross-linked polyethylene XLPE material from the inner, middle, and outer insulating layers with different operational years. The fourier transform infrared reflectance spectroscopy, thermogravimetric analysis, and differential scanning calorimetry were used to understand the relationship between the low-frequency dielectric and dynamic mechanical properties related to the operating years. Results show that the low-frequency dielectric and dynamic mechanical properties of the middle layer of the high-voltage XLPE insulating cables are better than those of the inner and outer layers. The crystallization peak temperature of the cable insulating material decreases with operating years. In our experiments, the restriction of molecular movement and volatilization of crosslinking byproducts during operation result in the improvement of properties for a set number of years.

Using our previously established xmrk transgenic zebrafish, hepatocellular carcinoma (HCC) was generated by induced expression of xmrk, which encoded a hyperactive epidermal growth factor receptor (EGFR) homolog, and regressed by suppression of xmrk expression. To investigate molecular changes in liver tumour progression and regression, RNA-Seq was performed for induced HCC and early and late stages of liver tissues during tumour regression. We found that Xmrk-induced zebrafish HCC shared strong molecular characteristics with a human HCC subtype (S2), which shows activated Myc signalling, upregulated phosphor-S6 and epithelial cell adhesion molecule. In the HCC stage, there were enhanced proteasome, antigen processing and presentation, aminosugars metabolisms, p53 and cell cycle pathways. During tumour regression, the transcriptomic profile showed a reversed trend of molecular changes compared with human HCC progression. Interestingly, distinct immune responses in tumour progression and regression were observed, including increased major histocompatibility complex class I (MHCI) at the HCC stage, enriched immune cell trafficking signals and inflammation in early regression and enhanced MHCII in late regression. Both neutrophils and macrophages were enriched during tumour progression and regression; however, the distribution of neutrophils and macrophages in HCC was relatively uniform, whereas both types of immune cells were regionally clustered during tumour regression, especially with dominant blood vessel association of macrophage in late regression, suggesting differential functions of these immune cells in tumour progression and regression. As tumour regression in our model resembles the targeted inhibition of EGFR in cancer therapy, our observations may provide molecular insights into the targeted inhibition and highlight the importance of immune response in tumour regression.

A novel tylophorine analog W-8 enhanced Foxp3 expression through both transcriptional and epigenetic programs, and its ameliorated TNBS induced colitis. Tylophorine and analogs are phenanthroindolizidine alkaloids, several of which have been reported to have anticancer, antiviral, and anti-inflammatory properties. However, their function in the immune system remains widely unknown. Transcription factor Foxp3 is critical for the development and function of Treg, which down-regulates the immune system and maintains tolerance to self-antigens. In the present study, we defined a novel tylophorine analog, W-8, enhanced TGF-β-induced Foxp3 expression at the mRNA and the protein levels. Interestingly, W-8 synergistically increased the level of TGF-β-induced p-Smad3 through inhibition of the AKT/mTOR pathway and enhanced the demethylation of the promoter region of the Foxp3 through inhibition of the ERK pathway and DNMT1 expression. Moreover, administration of W-8 suppressed TNBS-induced murine colitis and increased Tregs in lymphoid tissues. Finally, W-8 enhanced conversion of naïve T cells to Tregs in vivo. In summary, our results defined a novel compound that enhanced Foxp3 expression through transcriptional and epigenetic programs, and it might serve as a therapeutic agent for inflammatory diseases.

Manufacturing material resource planning (MMRP) increasingly involves complex sets of objectives and constraints; therefore traditional approaches typically result in a large monolithic model that is difficult to solve, understand, and maintain. The paper presents a multiagent-based constraint programming system, namely MRPlanner, which is aimed to solving large, particularly combinatorial, problems in MMRP. The system uses an asynchronous team (A-Team) architecture in which multiple problem solving agents cooperate with each other by exchanging results to produce a set of non-dominated solutions that show the tradeoffs between objectives. The agent-based approach employed in the system significantly improves the performance, reliability, intelligence and automation of constraint programming.

At present, image processing technology is more commonly applied to such fields as virtual reality and biological medicine, and permeates rapidly through agricultural scientific research. The image measuring method makes dairy cow type linear appraisal , which not only reduces workload but also gets rid of the shortcoming of the manual appraisal standard changing with human and time and the disadvantage of lacking impersonal impartiality. This paper describes a linear appraisal system for analyzing dairy cow types based on image processing, which consists of hardware design, software design, realization of characters click points operating and the interactive measurement of type character parameter for dairy cow. We use NI IMAQ Vision and NI Lab Windows/CVI 6.0 to develop the system software. The central purpose of image preconditioning is to clear up noise, and we use median filtering to eliminate noise in this approach.