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The first photoactivatable STING agonist, caged-diBSP01, is presented based on a novel and potent dimerized STING agonist diBSP01. Caged-diBSP01 allowed spatiotemporal control of STING activation by light, which was well characterized both in vitro and in vivo.

Figure S4. IRF8 reduces AR protein levels and activity.

IRF8 was not frequently mutated and methylated in clinical PCa tissues and cells.

In incurable castration-resistant prostate cancer (CRPC), resistance to the novel androgen receptor (AR) antagonist enzalutamide is driven mainly by AR overexpression. Here we report that the expression of interferon regulatory factor 8 (IRF8) is increased in primary prostate cancer but decreased in CRPC compared with normal prostate tissue. Decreased expression of IRF8 positively associated with CRPC progression and enzalutamide resistance. IRF8 interacted with AR and promoted its degradation via activation of the ubiquitin/proteasome systems. Epigenetic knockdown of IRF8 promoted AR-mediated prostate cancer progression and enzalutamide resistance in vitro and in vivo. Furthermore, IFNα increased expression of IRF8 and improved the efficacy of enzalutamide in CRPC by targeting the IRF8–AR axis. We also provide preliminary evidence for the efficacy of IFNα with hormonotherapy in a clinical study. Collectively, this study identifies IRF8 both as a tumor suppressor in prostate cancer pathogenesis and a potential alternative therapeutic option to overcome enzalutamide resistance.Significance:These findings identify IRF8-mediated AR degradation as a mechanism of resistance to AR-targeted therapy, highlighting the therapeutic potential of IFNα in targeting IRF8–AR axis in CRPC.

Therapeutic blockade of the programmed cell death protein-1 (PD-1) immune checkpoint pathways has resulted in significant reactivation of T cell-mediated antitumor immunity and is a promising clinical anticancer treatment modality in several tumor types, but the durable response rate remains relatively low (15%-20%) in most patients with HCC for unknown reasons. Evidence reveals that the interferon signaling pathway plays a critical role in modulating the efficacy and sensitivity of anti-PD-1 therapy against multiple tumor types, but the mechanisms are unclear.Using Kaplan-Meier survival analysis based on HCC databases, we found that deceased expression of interferon regulatory factor (IRF) 8 in HCC, among all the nine IRF members that regulate interferon signals, was associated with poor prognosis of patients with HCC. Moreover, gene set enrichment analysis identified the interferon-gamma and PD-1 signaling signatures as the top suppressed pathways in patients with IRF8-low HCC. Contrarily, overexpression of IRF8 in HCC cells significantly enhanced antitumor effects in immune-competent mice, modulating infiltration of tumor-associated macrophages (TAMs) and T cell exhaustion in tumor microenvironment. We further demonstrated that IRF8 regulated recruitment of TAMs by inhibiting the expression of chemokine (C-C motif) ligand 20 (CCL20). Mechanically, IRF8-mediated repression of c-fos transcription resulted in decreased expression of CCL20, rather than directly bound to CCL20 promoter region. Importantly, adeno-associated virus 8-mediated hepatic IRF8 rescue significantly suppressed HCC progression and enhanced the response to anti-PD-1 therapy.This work identified IRF8 as an important prognostic biomarker in patients with HCC that predicted the response and sensitivity to anti-PD-1 therapy and uncovered it as a therapeutic target for enhancing the efficacy of immune therapy.

Purpose: Exploring and studying the novel target of hepatocellular carcinoma (HCC) has been extremely important for its treatment. The principal objective of this project is to investigate whether myeloid derived growth factor (MYDGF) could accelerate the progression of HCC, and how it works. Methods: Cell proliferation, clonal formation, sphere formation and xenograft tumor experiments were used to prove the critical role of MYDGF in HCC progression. Tumor angiogenesis, immune cell infiltration, macrophage chemotaxis and inflammatory cytokines detection were utilized to clarify how MYDGF remodeled the tumor microenvironment (TME) to accelerate the progress of HCC. Results: Here, we reported a secretory protein MYDGF, which could be induced by hypoxia, was significantly upregulated in HCC and associated with poor clinical outcomes. Using bioinformatics and experimental approaches, we found that MYDGF promotes cell proliferation in vitro and in vivo through a mechanism that might involve enhanced self-renewal of liver CSCs. Furthermore, MYDGF can also promote tumor angiogenesis, induce macrophages to chemotaxis into tumor tissue, and then release various inflammatory cytokines, including IL-6 and TNF-α, which ultimately aggravate inflammation of tumor microenvironment and accelerate HCC progression. Conclusions: We provided evidence that MYDGF could directly affect the self-renewal of liver CSCs, and indirectly aggravate the inflammatory microenvironment to accelerate the progression of HCC.

The current knowledge about the effect of pCO2-driven ocean acidification on the bioaccumulation of pollutants in marine species is still scarce, as only limited types of pollutants have been investigated. Therefore, to obtain a better understanding of the effect of ocean acidification on the process of bioaccumulation and subsequent food safety, the accumulation of benzo[a]pyrene (B[a]P), chloramphenicol (CAP), and nitrofurazone (NFZ) in an edible bivalve species, Tegillarca granosa, under present and near-future ocean acidification scenarios was investigated in the present study. The health risks associated with consuming contaminated blood clams were also assessed using target hazard quotient (THQ), lifetime cancer risk (CR), or margin of exposure (MoE). To explain the alterations in bioaccumulation of these pollutants, the expressions of genes encoding corresponding key metabolic proteins were analyzed as well. The results obtained showed that ocean acidification exerted a significant effect on the accumulation of B[a]P, NFZ, and CAP in the clams. After four-week exposure to B[a]P, NFZ, or CAP contaminated seawater acidified with CO2 at pH 7.8 and 7.4, significantly greater amounts of B[a]P and lower amounts of NFZ and CAP were accumulated in the clams compared to that in the control. Although no non-carcinogenic risk of consuming B[a]P-contaminated blood clams was detected using the THQ values obtained, the CR values obtained indicated a high life-time risk in all groups. In addition, according to the MoE values obtained, the health risks in terms of consuming NFZ- and CAP-contaminated clams were significantly reduced under ocean acidification scenarios but still cannot be ignored, especially for children. The gene expression results showed that the ability of clams to eliminate B[a]P may be significantly constrained, whereas the ability to eliminate NFZ and CAP may be enhanced under ocean acidification scenarios, indicating that the changes in the accumulation of these pollutants may be due to the altered in vivo metabolism.

Hybrid electron beam lithography (EBL) and triple patterning lithography (TPL) is an advanced technology for IC manufacture. To solve the hybrid EBL and TPL layout decomposition problem (HETLD), we propose a discrete relaxation method in this paper. Existed works hard to evaluate the quality of the obtained results theoretically. Our discrete relaxation is the first approach which can be used to evaluate the solution quality. First, we introduce an extended minimum weight dominating set coloring problem (EMDSC). We show that the EMDSC problem is a discrete relaxation of the HETLD problem, which is solved by an integer linear programming (ILP) approach. Second, we legalize the relaxation solution to a feasible solution of the initial problem by inserting stitches and assigning EBLs. Experimental results show that our method achieves the near-optimal solution.

Quercetin, a typical flavonoid, possesses diverse biochemical and physiological actions, including antiplatelet, estrogenic, and anti-inflammatory properties. This review mainly centers on recent ten years findings with respect to intervening diabetes and its complications with the well-known flavonoid quercetin. After a short introduction of quercetin, major in vitro and in vivo findings are summarized showing that quercetin is a promising molecule for the treatment of these diseases. Finally, we contemplate future development and application prospects of quercetin. Despite the wealth of in animal research results suggesting the anti-diabetic and its complications potential of quercetin, its efficacy in diabetic human subjects is yet to be explored. The problem may become an important direction in the future research.

Diabetic retinopathy (DR) is the leading clinical cause of blindness in diabetic patients. Mitophagy participates in the pathogenesis of DR. Dynamin related protein 1 (Drp1) is associated with mitophagy. Here, we investigated whether Drp1 can regulate mitophagy to affect the progression of DR. We constructed DR rat model by administration of streptozocin. Primary rat retinal endothelial cells (RECs) were treated with high glucose (HG) as a DR cell model. Drp1 was highly expressed in the retinal tissues of DR rats and HG-treated RECs. Drp1 knockdown inhibited HG-mediated increase of reactive oxygen species (ROS) levels and apoptosis in RECs. Moreover, Drp1 silencing inhibited the expression of autophagy-related proteins LC3-II/LC3-1 and Beclin-1 and reduced LC3 puncta in HG-treated RECs. The expression of mitochondrial marker Tom20 was reduced and the levels of mitophagy were increased in the HG-treated RECs, which was rescued by Drp1 silencing. Drp1 knockdown repressed LC3-II expression in HG-treated RECs, indicating that autophagy flux was inhibited. Rapamycin (autophagy activator) enhanced ROS levels and apoptosis in HG-treated RECs by activating autophagy, which was rescued by Drp1 knockdown. In conclusion, these data demonstrated that Drp1 knockdown repressed apoptosis of rat retinal endothelial cells by inhibiting mitophagy. Thus, this work suggests that targeted regulation of Drp1 may become a treatment for DR.

Hepatic ischemia/reperfusion (I/R) injury is an inevitable complication of hepatic surgery occasioned by liver transplantation and resection. The progression from liver ischemia to reperfusion injury is accompanied by abnormal metabolism, Kupffer cell activation, neutrophil recruitment and the release of cytokines. Activation of several interferon regulatory factors (IRFs) has been reported to either enhance or restrict I/R progression, but the role of IRF8 in the regulation of I/R injury progression is still unknown. In this study, we explore the IRF8 function in the I/R-mediated liver injury using overexpressed hepatic IRF8 and knockout mice. According to our results, IRF8 knockout mice had significantly lower inflammatory cells infiltration, inflammatory cytokines release and serum aspartate aminotransferase/alanine aminotransferase levels that improved the necrotic injury after I/R, unlike the control mice. Conversely, the overexpression of IRF8 in WT mice markedly aggravated the liver structure damage and its abnormal function. We further showed that IRF8-mediated inflammatory cells infiltration were partly dependent on early autophagy and NF-κΒ signal pathway during I/R. AAV8-IRF8-I/R mice pretreated with autophagy inhibitor hydroxychloroquine and NF-κΒ signal pathway inhibitor secukinumab could drastically reverse the IRF8-mediated increase of neutrophil infiltration and chemokine release at different degrees. This work uncovered a critical role of IRF8 in the modulation of the hepatic microenvironment and as a potential target in the initial treatment of I/R injury.

Although judicious use of tyrosine kinase inhibitors that target BCR-ABL constitutes an effective strategy for the control of chronic myeloid leukemia (CML), drug resistance is observed due to kinase domain mutations, among which a major one is BCR-ABLT315I. In this study, we identified SHC004-221A1 as a potent inhibitor of T315I and other BCR-ABL mutants. Biochemical assays demonstrated that SHC004-221A1 has an inhibitory effect on all selected BCR-ABL mutants. In vitro studies showed that SHC004-221A1 inhibited the proliferation of tumor cell lines carrying native and T315I mutant BCR-ABL. Signaling pathway analysis revealed that SHC004-221A1 inhibited the phosphorylation of STAT5 and CrkL, which contributed to the apoptosis of CML cells. In vivo studies indicated that SHC004-221A1 suppressed BCR-ALBT315I-driven tumor growth in mice. Taken together, the results of this study suggested that SHC004-221A1 may be a promising BCR-ABLT315I inhibitor for the treatment of CML.

Persistent organic pollutants (POPs) are known to converge into the ocean and accumulate in the sediment, posing great threats to marine organisms such as the sessile bottom burrowing bivalves. However, the immune toxicity of POPs, such as B[a]P, under future ocean acidification scenarios remains poorly understood to date. Therefore, in the present study, the impacts of B[a]P exposure on the immune responses of a bivalve species, Tegillarca granosa, under present and future ocean acidification scenarios were investigated. Results obtained revealed an increased immune toxicity of B[a]P under future ocean acidification scenarios in terms of reduced THC, altered haemocyte composition, and hampered phagocytosis, which may attribute to the synergetic effects of B[a]P and ocean acidification. In addition, the gene expressions of pathogen pattern recognition receptors (TLR1, TLR2, TLR4, TLR6), pathway mediators (TRAF6, TAK1, TAB2, IKKα and Myd88), and effectors (NF-ĸB) of the important immune related pathways were significantly down-regulated upon exposure to B[a]P under future ocean acidification scenarios. Results of the present study suggested an increased immune toxicity of B[a]P under future ocean acidification scenarios, which will significantly hamper the immune responses of T. granosa and subsequently render individuals more susceptible to pathogens challenges.

Prostate cancer (PC) is the most diagnosed type of malignancy in men and the major frequently cause of cancer-related death worldwide. The androgen receptor (AR) has become a promising drug target for the treatment of PC. Here, we reported the design, optimization and evaluation of pyridine tetrahydroisoquinoline thiohydantoin derivatives with improved activity and safety as potent AR antagonists. The most promising compound 42f exhibited potent inhibitory activity on AR and strongly blocked AR nuclear translocation. Moreover, 42f displayed promising in vitro antitumor activity toward AR-dependent prostate cancer cell lines (LNCaP) and also demonstrated therapeutic effects in LNCaP xenograft tumor model in mice (TGI: 79%) with no apparent toxicity observed in vivo. More importantly, 42f showed negligible penetration of the brain-blood barrier (BBB) compared with enzalutamide. These results provide a foundation for the development of a new class of androgen receptor antagonists for potential therapeutics against PC with lower seizurogenic risk for patients.

In incurable castration-resistant prostate cancer (CRPC), resistance to the novel androgen receptor (AR) antagonist enzalutamide is driven mainly by AR overexpression. Here we report that the expression of interferon regulatory factor 8 (IRF8) is increased in primary prostate cancer but decreased in CRPC compared with normal prostate tissue. Decreased expression of IRF8 positively associated with CRPC progression and enzalutamide resistance. IRF8 interacted with AR and promoted its degradation via activation of the ubiquitin/proteasome systems. Epigenetic knockdown of IRF8 promoted AR-mediated prostate cancer progression and enzalutamide resistance in vitro and in vivo. Furthermore, IFNα increased expression of IRF8 and improved the efficacy of enzalutamide in CRPC by targeting the IRF8–AR axis. We also provide preliminary evidence for the efficacy of IFNα with hormonotherapy in a clinical study. Collectively, this study identifies IRF8 both as a tumor suppressor in prostate cancer pathogenesis and a potential alternative therapeutic option to overcome enzalutamide resistance. Significance: These findings identify IRF8-mediated AR degradation as a mechanism of resistance to AR-targeted therapy, highlighting the therapeutic potential of IFNα in targeting IRF8–AR axis in CRPC.

Erlotinib, an EGFR tyrosine kinase inhibitor has been introduced into cancer chemotherapy. However, the therapeutic effects of erlotinib in hepatocellular carcinoma (HCC) remain vaguely understood. Our previous study found that a hypoxia-mediated PLAGL2-EGFR-HIF-1/2α signaling loop in HCC decreased response to erlotinib. The current study has demonstrated that the combination of erlotinib and 2ME2 exerted synergistic antitumor effects against HCC. Further investigation showed that erlotinib increased the expression level of EGFR, HIF-2α, and PLAGL2, which contributes to the insensitivity of hypoxic HCC cells to erlotinib. The simultaneous exposure to 2ME2 effectively inhibited the expression level of EGFR, HIF-2α, and PLAGL2 that was induced by erlotinib. This contributes to the synergistic effect of the two therapeutic agents. Furthermore, the combination of erlotinib and 2ME2 induced apoptosis and inhibited the stemness of hypoxic HCC cells. Our findings potentially explain the mechanism of HCC insensitivity to erlotinib and provide a new strategy of combining EGFR and HIF1/2α inhibitors for HCC treatment.

Prostate cancer (PCa) is one of the most common malignancies affecting men worldwide. Androgen receptor (AR) has been a target of PCa treatment for nearly six decades. AR antagonists/degraders can effectively treat PCa caused by increased AR overexpression. However, all approved AR antagonists have similar chemical structures and exhibit the same mode of action on the protein. Although initially effective, resistance to these AR antagonists usually develops. Therefore, this calls for the identification of novel chemical structures of AR antagonists to overcome the resistance. Herein, we employed the synergetic combination of virtual and experimental screening to identify a flavonoid compound which not only effectively inhibits AR transcriptional activity, but also induces the degradation of the protein. Based on this compound, we designed and synthesized a series of derivatives. We discovered that the most potent compound 10e could effectively inhibit AR transcriptional activity, and possessed a profound ability to cause degradation of both full length- and ARv7 truncated forms of human AR. Notably, 10e efficiently inhibited the growth of ARv7 dependent prostate cancer cell-lines, which are completely resistant to all current anti-androgens. Compound 10e also showed strong antitumor activity in the LNCaP (androgen dependent prostate cancer cell line) in vivo xenograft model. These results provide a foundation for the development of a new class of AR antagonists.

MicroRNA-21 (miR-21) was reported to act as an oncogene during the development of many human tumors. However, little was revealed about the function of miR-21 in retinoblastoma (RB). In this study, we examined the expression of miR-21 in RB tissues and explored the relationship between miR-21 and phosphatase and tensin homolog (PTEN)/phosphatidylinositol-3-OH kinase (PI3K)/AKT signal. Quantitative real-time PCR (qRT-PCR) results showed that the level of miR-21 in RB tissues was higher than that in retinal normal tissues. In Weri-Rb-1 cells, miR-21 inhibitor suppressed the expression of miR-21 and cell viability, but improved cell apoptotic rates by modulating the levels of PDCD4, Bax, and Bcl-2. Meanwhile, miR-21 inhibitor suppressed cell migration and invasion via inhibiting the protein levels of MMP2 and MMP9 and significantly affected the expression of PTEN, PI3K, and p-AKT. Taken together, miR-21 inhibitor suppressed cell proliferation, migration, and invasion via the PTEN/PI3K/AKT signal. These findings revealed the molecular basis of miR-21 functioning in the progression of RB and provided a new means for cell therapy in RB.

Purpose: Psychologic depression has been shown to dysregulate the immune system and promote tumor progression. The aim of this study is to investigate how psychologic depression alters the immune profiles in prostate cancer. Experimental Design: We used a murine model of depression in Myc-CaP tumor-bearing immunocompetent FVB mice and Hi-myc mice presenting with spontaneous prostate cancer. Transwell migration and coculture assays were used to evaluate myeloid cell trafficking and cytokine profile changes evoked by Myc-CaP cells that had been treated with norepinephrine (NE), a major elevated neurotransmitter in depression. Chemoattractant, which correlated with immune cell infiltration, was screened by RNA-seq. The chemoattractant and immune cell infiltration were further confirmed using clinical samples of patients with prostate cancer with a high score of psychologic depression. Results: Psychologic depression predominantly promoted tumor-associated macrophage (TAM) intratumor infiltrations, which resulted from spleen and circulating monocytic myeloid-derived suppressor cell mobilization. Neuropeptide Y (NPY) released from NE-treated Myc-CaP cells promotes macrophage trafficking and IL6 releasing, which activates STAT3 signaling pathway in prostate cancer cells. Clinical specimens from patients with prostate cancer with higher score of depression revealed higher CD68+ TAM infiltration and stronger NPY and IL6 expression. Conclusions: Depression promotes myeloid cell infiltration and increases IL6 levels by a sympathetic-NPY signal. Sympathetic-NPY inhibition may be a promising strategy for patients with prostate cancer with high score of psychologic depression. See related commentary by Mohammadpour et al., p. 2363

Fil: Cheng, Yan. China Pharmaceutical University. Center for New Drug Safety Evaluation and Research. State Key Laboratory of Natural Medicines. Key Laboratory of Drug Discovery for Metabolic Disease; China Fil: Xing-Hua, Gao. China Pharmaceutical University. Center for New Drug Safety Evaluation and Research. State Key Laboratory of Natural Medicines. Key Laboratory of Drug Discovery for Metabolic Disease; China Fil: Li, Xian-Jing. China Pharmaceutical University. Center for New Drug Safety Evaluation and Research. State Key Laboratory of Natural Medicines. Key Laboratory of Drug Discovery for Metabolic Disease; China Fil: Cao, Qiu-Hua. China Pharmaceutical University. Center for New Drug Safety Evaluation and Research. State Key Laboratory of Natural Medicines. Key Laboratory of Drug Discovery for Metabolic Disease; China Fil: Zhao, Dan-Dan. China Pharmaceutical University. Center for New Drug Safety Evaluation and Research. State Key Laboratory of Natural Medicines. Key Laboratory of Drug Discovery for Metabolic Disease; China Fil: Zhou, Jin-Rong. Harvard Medical School. Beth Israel Deaconess Medical Center. Department of Surgery. Nutrition/Metabolism Laboratory; Estados Unidos Fil: Wu, Hong-Xi. China Pharmaceutical University. Center for New Drug Safety Evaluation and Research. State Key Laboratory of Natural Medicines. Key Laboratory of Drug Discovery for Metabolic Disease; China Fil: Wang, Yun. China Pharmaceutical University. Center for New Drug Safety Evaluation and Research. State Key Laboratory of Natural Medicines. Key Laboratory of Drug Discovery for Metabolic Disease; China Fil: You, Lin-Jun. China Pharmaceutical University. Center for New Drug Safety Evaluation and Research. State Key Laboratory of Natural Medicines. Key Laboratory of Drug Discovery for Metabolic Disease; China Fil: Yang, Hong-Bao. China Pharmaceutical University. Center for New Drug Safety Evaluation and Research. State Key Laboratory of Natural Medicines. Key Laboratory of Drug Discovery for Metabolic Disease; China Fil: He, Yun-Long. China Pharmaceutical University. Center for New Drug Safety Evaluation and Research. State Key Laboratory of Natural Medicines. Key Laboratory of Drug Discovery for Metabolic Disease; China Fil: Li, Yong-Ren. China Pharmaceutical University. Center for New Drug Safety Evaluation and Research. State Key Laboratory of Natural Medicines. Key Laboratory of Drug Discovery for Metabolic Disease; China Fil: Bian, Jin-Song. National University of Singapore. Yong Loo Lin School of Medicine. Department of Pharmacology; Singapur Fil: Zhu, Qing-Yi. Jiangsu Province Hospital of Traditional Chinese Medicine. Department of Urology; China Fil: Birnbaumer, Lutz. National Institute of Environmental Health Sciences. Neurobiology Laboratory; Estados Unidos Fil: Birnbaumer, Lutz. Pontificia Universidad Católica Argentina. Instituto de Investigaciones Biomédicas; Argentina Fil: Birnbaumer, Lutz. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Yang, Yong. China Pharmaceutical University. Center for New Drug Safety Evaluation and Research. State Key Laboratory of Natural Medicines. Key Laboratory of Drug Discovery for Metabolic Disease; China Abstract: Depression drives cancer progression and induces poor clinical outcome. However, the mechanisms underlying depression and cancer outcomes are unclear. In this work, we investigated 98 prostate cancer patients and found that patients with high score of psychological depression were correlated with tumor invasion and metastasis. We found focal adhesion kinase (FAK) was increased in cancer patients with metastatic features and high score of depression. FAK knockdown completely blocked depression-promoted tumor invasion in orthotopic transplantation tumors. In Hi-myc mice and a murine model of depression, sympathetic activation was detected in the prostate tissue. Further we showed that FAK activation was dependent on a cAMP-PKA signaling pathway. Our results demonstrated that the activation of a sympathetic-FAK signaling pathway in prostate cancer patients with high degrees of depression facilitates tumor invasion. We suggest that blocking β2AR with propranolol or inhibiting FAK activation with PF562 271 may be novel strategies for depressed patients with invasive prostate cancer.

An analytical method was developed for the simultaneous and rapid determination of seven microcystins in fish meat by liquid chromatography-tandem mass spectrometry (LC-MS/MS) coupled with pass-through solid phase extraction (SPE). The samples were extracted with methanol-water (90:10, v/v) after heat treatment by water bath at 80℃, and then cleaned up with an Oasis PRiME HLB pass-through SPE column. The samples were analyzed directly on a Waters XSelect HSS T3 column using 0.1% (v/v) aqueous formic acid and acidified acetonitrile (0.1% formic acid, v/v) as mobile phases. Qualitative and quantitative analysis of the analytes was carried out under the multiple reaction monitoring mode with positive electrospray ionization. The matrix matching external standard method was used for quantitation analysis. To solve the problem of parent ion selection of the microcystins, the ionization characteristics of microcystins were evaluated under different mobile phase conditions. Finally, the results showed that the acid could promote the intensity of the doubly charged ions significantly. The calibration curves were linear well in the corresponding concentration ranges, with correlation coefficient ≥ 0.99. The limits of quantification ranged from 0.30 to 2.0 μ g/kg. The average spiked recoveries for the seven microcystins were between 70.6% and 96.1% with the relative standard deviations of 3.4%-9.6%. The proposed method is sensitive, accurate, and efficient. It is applicable for the determination of microcystins in fish meat.

To evaluate the feasibility of transplantation of embryonic stem cell (ESC)-derived retinal cells in the treatment of retinal degeneration. Rat ESCs were isolated and induced into retinal progenitor cells (RPCs) in vitro, which were subsequently induced into retinal pigment epithelium cells (RPEs) and photoreceptors (PRCs). All cells were identified by Western blot detection of their specific markers. RPEs and PRCs were, respectively, injected into the retina of Royal College of Surgeons (RCSs) rats. Control group was injected with PBS. Post-transplantation visual function was determined by electroretinography (ERG). The histology of the whole eye was compared by H&E staining. RPEs and PRCs were successfully derived from rat ESCs through the two-step differentiation as indicated by the presence of ESC- (Oct-3/4, Nanog, TRA-1-60 and TRA-1-81), RPC- (Rx, Mitf, Pax6 and Chx10), RPE- (RPE65 and keratin) and PRC-specific markers (blue opsin, red/green opsin, recoverin and rhodopsin) in Western blot. The amplitude of ERG a- and b-wave in RPE- and PRC-transplanted groups at week 2 and 10 after transplantation was markedly higher compared with PBS controls. Retinal injury and vascular stress response was not detected in any of the RCS rats after transplantation. The developed stepwise protocol can derive retinal cells from ESCs. Transplantation of these retinal cells can restore visual function of RCS rats. Our study provides evidence for potential clinical application of ESC-based cell therapy for retinal degeneration.

Natural small peptides from foods have been proven to be efficient inhibitors of Angiotensin I–converting enzyme (ACE) for the regulation of blood pressure. The traditional ACE inhibitory peptides screening method is both time consuming and money costing, to the contrary, virtual screening method by computation can break these limitations. We establish a virtual screening method to obtain ACE inhibitory peptides with the help of Libdock module of Discovery Studio 3.5 software. A significant relationship between Libdock score and experimental IC50 was found, Libdock score = 10.063 log(1/IC50) + 68.08 (R2 = 0.62). The credibility of the relationship was confirmed by testing the coincidence of the estimated log(1/IC50) and measured log(1/IC50) (IC50 is 50% inhibitory concentration toward ACE, in μmol/L) of 5 synthetic ACE inhibitory peptides, which was virtual hydrolyzed and screened from a kind of seafood, Phascolosoma esculenta. Accordingly, Libdock method is a valid IC50 estimation tool and virtual screening method for small ACE inhibitory peptides. Practical Application Hypertension is the most important preventable risk factor for death worldwide. Antihypertension peptides from foods are natural resource with less side effect for hypertension treatment. Dipeptides and tripeptides, with higher in vivo bioavailability and less degradation in gastrointestinal tract, are potent Angiotensin I–converting enzyme inhibitors with high activity. By using virtual screening method in silico, these peptides can be efficiently acknowledged from foods, and sequentially harvested by directed large-scale hydrolysis.

Visible and shortwave-near-infrared spectroscopy was used to predict protein content in Spirulina powder. Three variable selection algorithms of partial least square based uninformative variable elimination, partial least square based genetic algorithm, and successive projections algorithm were analyzed. Results showed that it was necessary to operate partial least square based uninformative variable elimination before successive projections algorithm. Successive projections algorithm performed better than partial least square based genetic algorithm, because it had a better result and fewer selected variables. The results showed that partial least square based uninformative variable elimination-successive projections algorithm was a good hybrid algorithm for the multivariable selection, and it was feasible for using visible and shortwave-near-infrared spectroscopy to non-destructively estimate protein content in Spirulina powder.

Three sulphated polysaccharides, coded as BEMPA, BEMPB(1), BEMPB(2), were extracted from the mucilage of mud snail of Bullacta exarata and purified by DEAE-cellulose ion-exchange and size-exclusion chromatography. Structural analysis of purified polysaccharides by chemical and biochemical methods revealed BEMPA was a high (1→3,4)-linked mannose-containing polysaccharide with molecular weight of 22,977 Da. BEMPB(1), with molecular weight of 64,117 Da, was a high (1→3)-linked arabinose-containing polysaccharide. BEMPB(2) was mainly composed of (1→3,4)-linked mannose with molecular weight of 47,507Da. The comparison between sulphated polysaccharides and their desulphated products showed that sulphate substitutions of BEMPB(1) were deduced to be at the C-3 of (1→4)-linked mannose, while sulphate substitutions of BEMPA and BEMPB(2) were at C-4 of (1→3)-linked mannose. Furthermore, BEMPA exhibited highest inhibitory effects on growth of B-16 melanoma cells, and IC(50) were 31.1 μg/mL.

MicroRNA-21 (miR-21) was reported to act as an oncogene during the development of many human tumors. However, little was revealed about the function of miR-21 in retinoblastoma (RB). In this study, we examined the expression of miR-21 in RB tissues and explored the relationship between miR-21 and phosphatase and tensin homolog (PTEN)/phosphatidylinositol-3-OH kinase (PI3K)/AKT signal. Quantitative real-time PCR (qRT-PCR) results showed that the level of miR-21 in RB tissues was higher than that in retinal normal tissues. In Weri-Rb-1 cells, miR-21 inhibitor suppressed the expression of miR-21 and cell viability, but improved cell apoptotic rates by modulating the levels of PDCD4, Bax, and Bcl-2. Meanwhile, miR-21 inhibitor suppressed cell migration and invasion via inhibiting the protein levels of MMP2 and MMP9 and significantly affected the expression of PTEN, PI3K, and p-AKT. Taken together, miR-21 inhibitor suppressed cell proliferation, migration, and invasion via the PTEN/PI3K/AKT signal. These findings revealed the molecular basis of miR-21 functioning in the progression of RB and provided a new means for cell therapy in RB.

Computer assisted movement tracking was used to characterize the motility of two marine microalgae, Isochrysis galbana and Tetraselmis chui , and to investigate the toxicity of Cu, Pb, and Cd on motile percentage, curvilinear velocity, average path velocity, straight line velocity, linearity, straightness, and wobble. Except for motile percentage, all other motility parameters differed significantly between I. galbana and T. chui . Based on relative motile percentage data, the median effective concentration (EC 50 ) of Cu on the motility of I. galbana and T. chui was 31.4 and 1.3 μmol/L, respectively, while for Pb it was 37.8 and 10.9 μmol/L and for Cd it was 121.6 and 37.8 μmol/L, respectively. Compared to I. galbana , T. chui was more sensitive to all tested metals. The toxic effect of the heavy metals on motility exhibited the following decreasing order for both species: Cu > Pb > Cd. Our results indicate that I. galbana and T. chui motility is sensitive to heavy metals and can be used as an indicator for toxicology bioassays.

The authentication of food products is critically important in a global economy in public-health and economic terms. The specific aims of this study were to evaluate the application of full-spectrum and NIR spectroscopy and to evaluate the adoption of PLS and LS-SVM models to accomplish a rapid and non-invasive quantification of the two common adulterants, flour and mungbean powder, in Spirulina powder. The results showed that, using all treatment sets, only the LS-SVM models were adequate in predicting either adulterant under both full spectra and NIR spectra. The use of NIR spectra would allow detection of adulterants even when masked by food dyes. Design value analysis indicated that the benefits per unit cost of applying the NIR spectra to quantify adulterants in Spirulina powder significantly exceeded that of using full spectra, and that the value of employing the LS-SVM models under NIR spectra exceeded that of using the PLS models.

The relationship between growth rates of algae and structure parameters of closed flat-panel photobioreactors was investigated. Ad/Ar (cross-section area of the downcomer/cross-section area of the riser), h0 (clearance from the upper edge of the baffles to the water level), and h1 (clearance from the lower edge of the baffles to the bottom of the reactor) were selected as the inner structure parameters. CFD (Computational Fluid Dynamics) was used to simulate the hydrodynamic parameter (TKE)d (turbulence kinetic energy of the downcomers) and the secondary parameters e (ratio between td and tc), tc (cycle time), and DZ (dead zones), which were deduced from the hydrodynamic parameters mentioned above. The effects of (TKE)d, e, tc, DZ, and the inner structure parameters on cell growth of Isochrysis galbana 3011 were analyzed using data collected in 15-L airlift flat-panel photobioreactors. A model was developed to predict algae cell growth based on these inner structure parameters, thereby providing a new method for photobioreactor optimization. Validity of the model was confirmed by experimental data of I. galbana 3011 cultured in 15-L and 300-L photobioreactors, respectively. Finally, the prospect of applying CFD to photobioreactor optimization was discussed.

Sipuncula are of increasing interest for ¢sheries and aquaculture in China. Sustainable harvests will rely on a better knowledge of reproductive characteristics and stock enhancement. Here, we investigated the structural characteristics of and seasonal changes in germ cell development of the sipunculid Phascolosoma esculenta from the south-eastern coast of Zhejiang, China. An annual survey of egg numbers in the coelom (body cavity) £uid by light and electron microscopy of the females indicates that P. esculenta is dioecious. No de¢ned gonad but dissociated germ cells were found in the coelomic cavity during the 1-year observation. The germ cells showed multiplication and development in the coelomic cavity. Reproduction took place from May to September, with a peak in July and August. The oogenesis can be divided into four phases: cell proliferation, pre-vitellogenesis, vitellogenesis and egg envelope formation and maturation. The process of spermatogenesis can also be divided into four phases: cell multiplication, cell growth, cell maturation and metamorphosis. Monthly changes in the relative number of eggs in each stage indicate that P. esculenta lays eggs in batches. The sperm thrives in the coelomic £uid in the form of cell groups with patterns of genesis and release similar to those of the eggs. Eggs of P. esculenta were fertilized only when reaching the nephridium. The sex ratio was about 1:1 throughout the year.

To change the traditional management of agricultural production, using ZigBee technology for short distance wireless transmission to design intelligent farmland environment remote monitoring system, which integrated communication, computer and network all aspects of technology. The real-time, accurate data collection of farmland soil PH value, temperature and humidity surrounding the plant, light intensity, crop growth and bacteria occur posture, provide reliable data for the intelligent agricultural production, thereby increasing the level of intelligence of agricultural management, and promote modernization of agricultural production process.

Natural small peptides from foods have been proven to be efficient inhibitors of Angiotensin I-converting enzyme (ACE) for the regulation of blood pressure. The traditional ACE inhibitory peptides screening method is both time consuming and money costing, to the contrary, virtual screening method by computation can break these limitations. We establish a virtual screening method to obtain ACE inhibitory peptides with the help of Libdock module of Discovery Studio 3.5 software. A significant relationship between Libdock score and experimental IC(50) was found, Libdock score = 10.063 log(1/IC(50)) + 68.08 (R(2) = 0.62). The credibility of the relationship was confirmed by testing the coincidence of the estimated log(1/IC(50)) and measured log(1/IC(50)) (IC(50) is 50% inhibitory concentration toward ACE, in μmol/L) of 5 synthetic ACE inhibitory peptides, which was virtual hydrolyzed and screened from a kind of seafood, Phascolosoma esculenta. Accordingly, Libdock method is a valid IC(50) estimation tool and virtual screening method for small ACE inhibitory peptides.

Computer assisted movement tracking was used to investigate the toxicity effect of commonly used aquiculture disinfectants (potassium permanganate and bleaching powder) on the motility of two marine microalgae Isochrysis galbana and Tetraselmis chui. Both disinfectants tested showed significant effect (p 50 of potassium permanganate on motility of I. galbana and T. chui was 0.85 and 5.00mg/L, respectively. For bleaching powder, it was 4.98 and 5.09mg/L for I. galbana and T. chui, respectively. Compared to T. chui, I. galbana was more sensitive to Potassium permanganate. Our results reveal a dose-response relationship between tested disinfectants and motility characters and indicate that the motility of I. galbana and T. chui is sensitive to disinfectants tested and can be used as an indicator for toxicology bioassays.

Ultrastructural characteristics of coelomocytes of the sipunculid Phascolosoma esculenta were studied by transmission electron microscopy. There are several cell types in the coelomic fluid, including three kinds of granulocytes, vesicular cells, germ cells, amoebocytes, phagocytes, and erythrocytes; there are also a new cell complex which is composed of podocytes and granular cells. And several other cell types (erythrocyte and different kinds of granulocytes) gathering together was discovered in the coelomic fluid of P. esculenta. Functional interpretations were provided for these cells using morphological evidence. The coelomocytes from different sipunculid genera and Annelida were compared. The structural diversity of coelomocytes provides both taxonomic characteristics for discriminative identification and phylogenetic markers in Phascolosoma and other sipunculid taxa.

In order to change the traditional management of agricultural production, ZigBee technology is used in short distance wireless transmission to design intelligent farmland environment remote monitoring system, which integrates communication, computer and all aspects of network technology. The real-time, accurate data collection of farmland soil pH value, the temperature and humidity surroundings of the plants, illumination intensity and crop chlorophyll content, all will provide reliable data for intelligent agricultural production, thereby, the level of intelligence of agricultural management is increased. Based on precision guidance, irrigation will become intelligent which can avoid the waste of water resources and low utilisation rate caused by free operation. At the same time, it will promote modernisation of agricultural production process.

Recent studies have investigated anti-hypertensive peptides derived from natural food products. In this work, we focus on Phascolosoma esculenta as a resource of anti-hypertensive peptides, which is also a seafood with high nutritive value. Compared with FAO/WHO requirements, P. esculenta was confirmed to contain high contents of amino acids and minerals. To investigate the anti-hypertensive activity of P. esculenta, water-soluble and insoluble proteins were extracted and hydrolysed by pepsin and trypsin, respectively. The hydrolysates of water-soluble proteins derived by pepsin and pepsin-trypsin exhibited angiotensin-converting enzyme (ACE) inhibitory activity with IC50 values of 0.67 and 0.24 mg ml(-1), respectively, and those of water-insoluble proteins presented IC50 values of 0.4 and 0.1 mg ml(-1), respectively. Experiments on 'spontaneously hypertensive rats' (SHRs) were carried out to test the anti-hypertension activity in vivo, which confirmed that the hydrolysates played a significant role in reducing both diastolic blood pressure (DBP) and systolic blood pressure (SBP). Consistently, the in vivo anti-hypertensive activity of the hydrolysis products of pepsin and trypsin used together was also higher than that by using pepsin hydrolysis products alone. As stated in both sets of results, we believe that P. esculenta is an excellent resource of antihypertensive peptides and warrants further investigation.