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Developing effective treatments for colorectal cancers through combinations of small-molecule approaches and immunotherapies present intriguing possibilities for managing these otherwise intractable cancers. During a broad-based, screening effort against multiple colorectal cancer cell lines, we identified indole-substituted quinolines (ISQ), such as N7,N7-dimethyl-3-(1-methyl-1H-indol-3-yl)quinoline-2,7-diamine (ISQ-1), as potent in vitro inhibitors of several cancer cell lines. We found that ISQ-1 inhibited Wnt signaling, a main driver in the pathway governing colorectal cancer development, and ISQ-1 also activated adenosine monophosphate kinase (AMPK), a cellular energy–homeostasis master regulator. We explored the effect of ISQs on cell metabolism. Seahorse assays measuring oxygen consumption rate (OCR) indicated that ISQ-1 inhibited complex I (i.e., NADH ubiquinone oxidoreductase) in the mitochondrial, electron transport chain (ETC). In addition, ISQ-1 treatment showed remarkable synergistic depletion of oncogenic c-Myc protein level in vitro and induced strong tumor remission in vivo when administered together with BI2536, a polo-like kinase-1 (Plk1) inhibitor. These studies point toward the potential value of dual drug therapies targeting the ETC and Plk-1 for the treatment of c-Myc–driven cancers.

Cancer cells undergo significant “metabolic remodeling” to provide sufficient ATP to maintain cell survival and to promote rapid growth. In colorectal cancer cells, ATP is produced by mitochondrial oxidative phosphorylation and by substantially elevated cytoplasmic glucose fermentation (i.e., the Warburg effect). Glucose transporter 1 (GLUT1) expression is significantly increased in colorectal cancer cells, and GLUT1 inhibitors block glucose uptake and hence glycolysis crucial for cancer cell growth. In addition to ATP, these metabolic pathways also provide macromolecule building blocks and signaling molecules required for tumor growth. In this study, we identify a diaminobutoxy-substituted isoflavonoid (DBI-1) that inhibits mitochondrial complex I and deprives rapidly growing cancer cells of energy needed for growth. DBI-1 and the GLUT1 inhibitor, BAY-876, synergistically inhibit colorectal cancer cell growth in vitro and in vivo. This study suggests that an electron transport chain inhibitor (i.e., DBI-1) and a glucose transport inhibitor, (i.e., BAY-876) are potentially effective combination for colorectal cancer treatment.

Supplementary Data from Diaminobutoxy-substituted Isoflavonoid (DBI-1) Enhances the Therapeutic Efficacy of GLUT1 Inhibitor BAY-876 by Modulating Metabolic Pathways in Colon Cancer Cells

Developing effective treatments for colorectal cancers through combinations of small-molecule approaches and immunotherapies present intriguing possibilities for managing these otherwise intractable cancers. During a broad-based, screening effort against multiple colorectal cancer cell lines, we identified indole-substituted quinolines (ISQ), such as N7,N7-dimethyl-3-(1-methyl-1H-indol-3-yl)quinoline-2,7-diamine (ISQ-1), as potent in vitro inhibitors of several cancer cell lines. We found that ISQ-1 inhibited Wnt signaling, a main driver in the pathway governing colorectal cancer development, and ISQ-1 also activated adenosine monophosphate kinase (AMPK), a cellular energy–homeostasis master regulator. We explored the effect of ISQs on cell metabolism. Seahorse assays measuring oxygen consumption rate (OCR) indicated that ISQ-1 inhibited complex I (i.e., NADH ubiquinone oxidoreductase) in the mitochondrial, electron transport chain (ETC). In addition, ISQ-1 treatment showed remarkable synergistic depletion of oncogenic c-Myc protein level in vitro and induced strong tumor remission in vivo when administered together with BI2536, a polo-like kinase-1 (Plk1) inhibitor. These studies point toward the potential value of dual drug therapies targeting the ETC and Plk-1 for the treatment of c-Myc–driven cancers.

An efficient method for regioselective synthesis of C-7 Mannich bases of 6-hydroxyaurones was accomplished by the N,N-dialkylaminomethylation using aminals prepared from dimethylamine, dipropylamine, bis(2-methoxyethyl)amine, N-methylbutylamine, N-methylbenzylamine, morpholine, piperidine, and 1-methylpiperazine. Further transformation of 7-(N,N-dialkylamino)methyl group in these aurones led to formation of C-7 acetoxymethyl and methoxymethyl derivatives of 6-hydroxyaurones, some of which showed promising inhibition of PC-3 prostate cancer cell proliferation in the high nanomolar to low micromolar range that exceeded that of cisplatin. Compound 12c (R3 = Ac, Ar = 3,4-OMePh) displays 75% inhibition of PC-3 prostate cancer cells proliferation at 300 nM concentration.

Temporal data are ubiquitous nowadays and efficient management of temporal data is of key importance. A temporal data typically describes the evolution of an object over time. One of the most useful queries over temporal data are the durable top-k queries. Given a time window, a durable top-k query finds the objects that are frequently among the best. Existing solutions to durable top-k queries assume that all temporal data are sampled at the same time points (i.e., at any time, there is a corresponding observed value for every temporal data). However, in many practical applications, temporal data are collected from multiple data sources with different sampling rates. In this light, we investigate the efficient processing of durable top-k queries over temporal data with different sampling rates.We propose an efficient sweep line algorithm to process durable top-k queries over non-synchronized temporal data. We conduct extensive experiments on two real datasets to test the performance of our proposed method. The results show that our methods outperforms the baseline solutions by a large margin.

Cancer cells undergo significant “metabolic remodeling” to provide sufficient ATP to maintain cell survival and to promote rapid growth. In colorectal cancer cells, ATP is produced by mitochondrial oxidative phosphorylation and by substantially elevated cytoplasmic glucose fermentation (i.e., the Warburg effect). Glucose transporter 1 (GLUT1) expression is significantly increased in colorectal cancer cells, and GLUT1 inhibitors block glucose uptake and hence glycolysis crucial for cancer cell growth. In addition to ATP, these metabolic pathways also provide macromolecule building blocks and signaling molecules required for tumor growth. In this study, we identify a diaminobutoxy-substituted isoflavonoid (DBI-1) that inhibits mitochondrial complex I and deprives rapidly growing cancer cells of energy needed for growth. DBI-1 and the GLUT1 inhibitor, BAY-876, synergistically inhibit colorectal cancer cell growth in vitro and in vivo. This study suggests that an electron transport chain inhibitor (i.e., DBI-1) and a glucose transport inhibitor, (i.e., BAY-876) are potentially effective combination for colorectal cancer treatment.

Artificial intelligence ultrasound of prostate (AIUSP)-targeted biopsy has been used for prostate cancer (PCa) diagnosis. The objective of this prospective multi-center head-to-head clinical randomized comparative trail (RCT) is to compare PCa detection rate in the TRUS-guided 12-core standard systematic biopsy (TRUS-SB) group and cognitive fused mpMRI-guided 12-core biopsy (mpMRI) group against AIUSP group.Four hundred patients were randomized to three arms and underwent biopsies by TRUS-SB (n = 133), mpMRI (n = 134), and AIUSP (n = 133) between January 2015 and December 2017. In TRUS-SB group, a standard 12-core systematic biopsy was performed. In mpMRI group, mpMRI-suspicious lesions (PI-RADS 3-5) were targeted by 2-core biopsy followed by a 10-core systematic biopsy. Otherwise, 12-core systematic biopsy was performed. In AIUSP group, a 6-core targeted biopsy was performed. The primary endpoint was PCa detection rate.AIUSP detected the highest rate of PCa (66/133, 49.6%) compared to TRUS-SB (46/133, 34.6%, p = 0.036) and mpMRI (48/134, 35.8%, p = 0.052). Compared to TRUS-SB (35/133, 26.3%) and mpMRI (31/134, 23.1%) groups, clinically significant PCa (csPCa) detection rate was 32.3% (43/133) in AIUSP group. Overall biopsy core positive rate in the TRUS-SB group (11.0%, 176/1598) and in the mpMRI group (12.7%, 204/1608) was significantly lower than that in the AIUSP group (22.7%, 181/798, p 0.001).AIUSP detected the highest rate of overall and significant PCa compared to TRUS-SB and mpMRI, and could be used as an alternative to systematic biopsy in the future.This trial was registered in ISRCTN (ISRCTN18033113).

The COVID-19 pandemic has adversely impacted the agricultural supply chain, export of agricultural products, and overall food security. However, minimal exploration has been attempted of farmers’ confidence in agricultural production recovery after the COVID-19 pandemic. Therefore, this study intends to explore the determinants of farmers’ confidence in agricultural production recovery in China during the early stages of the COVID-19 pandemic. More specifically, we analyzed the relationship between risk expectation and social support on the farmers’ confidence in agricultural production recovery by using the ordered probit model. Cross-sectional survey data were collected from February to March 2020 from 458 farm households in the 7 provinces of China to produce the findings. We found that the risk expectation of farmers had a significant negative impact on farmers’ confidence in agricultural production recovery. Social support seemingly had a significant positive impact on the farmers’ confidence in agricultural production recovery, and could play a supportive role in moderating the relationship between risk expectation and farmers’ confidence in recovery. However, social support alleviates the adverse effect of risk expectation on farmers’ confidence in agricultural production recovery to a certain extent. In addition, there were intergenerational differences in the effects of risk expectation and social support on farmers’ confidence in agricultural production recovery. These results imply that policies establishing the risk early warning mechanisms for agricultural production and strengthening the social support from governments and financial institutions are likely to significantly impact agricultural development in the post-COVID-19 era. The formal and informal risk minimization mechanisms should extend their support to vulnerable sectors such as agribusiness.

Limited evidence supports the efficacy of iron-rich foods (IRFs) in improving iron status during pregnancy.The study aims to evaluate the effect of IRFs on iron status and biomarkers of iron metabolism in the third trimester of pregnancy.A total of 240 pregnant women at 11-13 wk of gestation without iron-deficiency anemia (IDA) in South China were recruited to this single-blind clinical trial [non-IDA referred to both hemoglobin (Hb) ≥110g/L and serum ferritin (SF) ≥15ng/mL], randomly assigned to 1) control, 2) IRFs containing 20 mg iron/d (IRF-20), or 3) IRFs containing 40 mg iron/d (IRF-40). The IRFs were consumed 3 days a week, including pork liver, chicken/duck blood, soybean, and agaric. The IRFs started at recruitment and ended in the predelivery room. Primary outcome included anemia (Hb 110 g/L), iron deficiency (ID, definition 1: SF 15 ng/mL; definition 2: SF 12 ng/mL), and IDA (ID and Hb 110 g/L). Secondary outcome was plasma Hb and iron indices, including SF, serum hepcidin, and iron.All participants who completed the trial with full data (n = 170) were included in the analysis. At the endline, both intervention groups showed lower ID and IDA rates than control. Specifically, IRF-40 showed a lower ID (SF 12 ng/mL) rate than control (9.0% compared with 22.8%, P = 0.022). For IDA by definition 1, the incidence in IRF-40 was lower than that in control (1.9% compared with 8.9%, P = 0.045). For IDA by definition 2, the incidence in IRF-20 was lower than that in control (3.9% compared with 17.9%, P = 0.049). Moreover, IRF-20 showed higher SF concentrations than control (P = 0.039). No effects of IRFs on anemia (P = 0.856), plasma Hb (P = 0.697), serum hepcidin (P = 0.311), and iron (P = 0.253) concentrations were observed. The assessed iron intakes were 22.2 mg/d in IRF-20 and 25.0 mg/d in IRF-40, respectively.Antenatal IRFs reduce the risk of ID and IDA in late pregnancy, although the present results are inadequate to confirm an ideal dosage (No. ChiCTR1800017574).

Structure-activity relationships (SAR) in the aurone pharmacophore identified heterocyclic variants of the (Z)-2-benzylidene-6-hydroxybenzofuran-3(2H)-one scaffold that possessed low nanomolar in vitro potency in cell proliferation assays using various cancer cell lines, in vivo potency in prostate cancer PC-3 xenograft and zebrafish models, selectivity for the colchicine-binding site on tubulin, and absence of appreciable toxicity. Among the leading, biologically active analogs were (Z)-2-((2-((1-ethyl-5-methoxy-1H-indol-3-yl)methylene)-3-oxo-2,3-dihydrobenzofuran-6-yl)oxy)acetonitrile (5a) and (Z)-6-((2,6-dichlorobenzyl)oxy)-2-(pyridin-4-ylmethylene)benzofuran-3(2H)-one (5b) that inhibited in vitro PC-3 prostate cancer cell proliferation with IC50 values below 100 nM. A xenograft study in nude mice using 10 mg/kg of 5a had no effect on mice weight, and aurone 5a did not inhibit, as desired, the human ether-à-go-go-related (hERG) potassium channel. Cell cycle arrest data, comparisons of the inhibition of cancer cell proliferation by aurones and known antineoplastic agents, and in vitro inhibition of tubulin polymerization indicated that aurone 5a disrupted tubulin dynamics. Based on molecular docking and confirmed by liquid chromatography-electrospray ionization-tandem mass spectrometry studies, aurone 5a targets the colchicine-binding site on tubulin. In addition to solid tumors, aurones 5a and 5b strongly inhibited in vitro a panel of human leukemia cancer cell lines and the in vivo myc-induced T cell acute lymphoblastic leukemia (T-ALL) in a zebrafish model.

Mitochondria are ideal anti-tumor target due to mitochondria's central regulation role in cell apoptosis and tumor resistance to apoptosis. There are several challenges for mitochondrial targeting drug delivery, including complex multistep preparations, low drug- loading and systemic toxicity from the carriers. To address these issues, we firstly constructed mitochondria-targeting nanodrugs self-assembled from 9-O-octadecyl substituted berberine derivative (BD) using simple nano-precipitation approach. BD-based nanodrugs were modified by DSPE-PEG2000 (distearylphosphatidylethanolamine- methoxypolyethylene glycol 2000) to increase stability. Negatively charged hyaluronic acid (HA) was further coated to conceal positive charges and achieve tumor targeting. PEG and HA dually modified BD NDs (HA/PEG/BD NDs) were prepared with surface charge of −25.8 mV and high drug loading >70%. The degradation of HA by hyaluronidase (HAase) at tumor tissue allowed the exposure of the positively charged PEG/BD NDs to the cells, which is beneficial for cell uptake and further lysosome escape and mitochondrial targeting. Then, HA/PEG/BD NDs were investigated to induce apoptosis through dissipating mitochondria membrane potential, releasing cytochrome C, increasing the activities of caspase 9/3, activating the pro-apoptotic Bax, suppressing the anti-apoptotic Bcl-2 and upregulating ROS levels. In the A549 xenografted tumor model, HA/PEG/BD NDs exhibited obvious tumor cell mitochondrial targeting and significant anti-tumor efficacy. Overall, comparing to conventional nanoparticles, mitochondrial targeting HA/PEG/BD NDs provide a new strategy for cancer treatment with enhanced drug-loading, relatively simplified preparation processes and reduced carrier toxicities.

There has been an increased interest in the production of sustainable biochar in the past years, as biochar shows versatile physicochemical properties and, can have a wide applicability in diverse fields. Comprehensive studies have been made to characterize biochar produced from various biomass materials, using different production technologies and under different process conditions. However, research is still lacking in correlating biochar properties needed for certain applications with (i) feedstock, (ii) biochar production processes and conditions and (iii) biochar upgrading and modification strategies. To produce biochar with desired properties, there is a great need to establish and clarify such correlations, which can guide the selection of feedstock, tuning and optimization of the production process and more efficient utilization of biochar. On the other hand, further elucidation of these correlations is also important for biochar-stakeholder and end-users for predicting physiochemical properties of biochar from certain feedstock and production conditions, assessing potential effects of biochar utilization and clearly address needs towards biochar critical properties. This review summarizes a wide range of literature on the impact of feedstocks and production processes and reactions conditions on the biochar properties and the most important biochar properties required for the different potential applications. Based on collected data, recommendations are provided for mapping out biochar production for different biochar applications. Knowledge gaps and perspectives for future research have also been identified regarding the characterization and production of biochar.

The importance of upregulated Wnt signaling in colorectal cancers led to efforts to develop inhibitors that target β-catenin in this pathway. We now report that several “Wnt inhibitors” that allegedly target β-catenin actually function as mitochondrial proton uncouplers that independently activate AMPK and concomitantly inhibit Wnt signaling. As expected for a process in which mitochondrial uncoupling diminishes ATP production, a mitochondrial proton uncoupler, FCCP, and a glucose metabolic inhibitor, 2-DG, activated AMPK and inhibited Wnt signaling. Also consistent with these findings, a well-known “Wnt inhibitor”, FH535, functioned as a proton uncoupler, and in support of this finding, the N-methylated analog, 2,5-dichloro-N-methyl-N-(2-methyl-4-nitrophenyl)benzenesulfonamide (FH535-M) was inactive as an uncoupler and Wnt inhibitor. Apart from suggesting an opportunity to develop dual Wnt inhibitors and AMPK activators, these findings provide a cautionary tale that claims for Wnt inhibition alone require scrutiny as possible mitochondrial proton uncouplers or inhibitors of the electron transport chain.

Fluorinated, phenylethynyl-substituted heterocycles that possessed either an N-methylamino or N,N-dimethylamino group attached to heterocycles including pyridines, indoles, 1H-indazoles, quinolines, and isoquinolines inhibited the proliferation of LS174T colon cancer cells in which the inhibition of cyclin D1 and induction of the cyclin-dependent kinase inhibitor-1 (i.e., p21Wif1/Cip1) served as a readout for antineoplastic activity at a cellular level. On a molecular level, these agents, particularly 4-((2,6-difluorophenyl)ethynyl)-N-methylisoquinolin-1-amine and 4-((2,6-difluorophenyl)ethynyl)-N,N-dimethylisoquinolin-1-amine, bound and inhibited the catalytic subunit of methionine S-adenosyltransferase-2 (MAT2A).

The factors affecting intracellular ice formation (IIF) and growth is essential to the mechanistic understanding of cellular damage through freezing. In the aid of high speed and high-resolution cryo-imaging technology, the broad bean intracellular ice formation and growth processes were successfully captured during freezing. Cytochalasin B（CB）was used to solubilize the cytoskeleton. Images of IIF were compared between cells with and without cytoskeleton. The behavior of intracellular ice crystal formation in plant tissues with or without CB was evaluated using changes of cell areas, the probability of crystallization, and growing rate of intracellular ice crystal. Moreover, light intensity figures were used to determine cell damage. This study showed that the cytoskeleton was involved in ice crystal nucleation mechanism during freezing responses of the plant cells.

Innovative imaging modalities are applied for diagnosing and follow-up of prostate cancer (PCa). To perform authentic targeted biopsies and evaluate prostate changes, it is essential to reliably identify and hit targets in their true anatomical location over time. A newly described image correlation method (Internal Fusion) allows precise correlation of serial transrectal ultrasound (TRUS) images in vitro and vivo. Identifiable morphologic structures such as prostatic calcifications define internal anchor points (Internal Landmarks) to facilitate exact correlation. After verification of Internal Fusion in vitro, 83 patients were included from January to August 2017 with at least one prior series of 5-mm spaced TRUS images stored as computed tomographic (US-CT) online data set. Two experienced operators collected new images in correlation with each cross-section of the previous examinations based on Internal Landmarks. Of the 83 patients, fifty (60%) had prior negative biopsies. Fourteen (17%) wished US-CT targeted biopsies and PCa were detected in 7/14 patients (50%). Overall, accurate imaging correlations were attained in 369/397 slices (93%). In initial and repeat biopsy, 31/31 and 35/35 images could be correlated detecting PCa in 5/8 (63%) and 2/6 patients (33%). The longest observation of prostate changes over time (Trend Monitoring) with accurate image correlation was 8 years and 6 months. Internal Fusion by Internal Landmarks ensures exact correlation in long-term follow-up. It is possible to precisely monitor trends in prostate tissue changes. In case of PCa suspicion, biopsies could be targeted with high accuracy by Internal Fusion, even over time.

Cytisine-linked isoflavonoids (CLIFs) inhibited PC-3 prostate and LS174T colon cancer cell proliferation by inhibiting a peroxisomal bifunctional enzyme. A pull-down assay using a biologically active, biotin-modified CLIF identified the target of these agents as the bifunctional, peroxisomal enzyme, hydroxysteroid 17β-dehydrogenase-4 (HSD17B4). Additional studies with truncated versions of HSD17B4 established that CLIFs specifically bind the C-terminus of HSD17B4 and selectively inhibited the enoyl CoA hydratase but not the D-3-hydroxyacyl CoA dehydrogenase activity. HSD17B4 was overexpressed in prostate and colon cancer tissues, knocking down HSD17B4 inhibited cancer cell proliferation, suggesting that HSD17B4 is a potential biomarker and drug target and that CLIFs are potential probes or therapeutic agents for these cancers.

Summary Aberrant activation of Wnt signaling triggered by mutations in either Adenomatous Polyposis Coli (APC) or CTNNB1 (β-catenin) is a hallmark of colorectal cancers (CRC). As part of a program to develop epigenetic regulators for cancer therapy, we developed carboxamide-substituted benzhydryl amines (CBAs) bearing either aryl or heteroaryl groups that selectively targeted histone lysine demethylases (KDMs) and functioned as inhibitors of the Wnt pathway. A biotinylated variant of N-((5-chloro-8-hydroxyquinolin-7-yl) (4-(diethylamino)phenyl)-methyl)butyramide (CBA-1) identified KDM3A as a binding partner. KDM3A is a Jumonji (JmjC) domain-containing demethylase that is significantly upregulated in CRC. KDM3A regulates the demethylation of histone H3's lysine 9 (H3K9Me2), a repressive marker for transcription. Inhibiting KDM3 increased H3K9Me2 levels, repressed Wnt target genes, and curtailed in vitro CRC cell proliferation. CBA-1 also exhibited in vivo inhibition of Wnt signaling in a zebrafish model without displaying in vivo toxicity., Graphical Abstract, Highlights • A class of carboxamide-substituted benzhydryl amine (CBA) Wnt inhibitors • A biological active, biotinylated CBA to identify KDM3A as a direct target • CBA-1 interacted with the Mn2+ ion in the JmjC domains of KDM3A/3B • CBA-1 inhibited Wnt signaling in colon cancer cells and in zebrafish models, Biological Sciences; Biochemistry; Biochemical Mechanism

Growth hormone (GH) is an anabolic hormone that regulates linear growth during critical developmental stages, and maintains healthy metabolism throughout life. The ultradian rhythm of GH secretion is orchestrated by intricate actions of hypothalamic stimulatory growth hormone-releasing hormone (GHRH) and inhibitory somatostatin (SST), acting on somatotrophs in the anterior pituitary gland (APG). The interplay between GHRH and SST generates the pulsatile pattern of GH release. This patterned release is defined by the amount of GH secreted per burst (amplitude of GH release) and the secretory pulse frequency and regularity (as denoted by pulse number and approximate entropy (ApEn); defined using deconvolution analysis). The patterned release of GH is entrained to meet physiological demands, with numerous central and peripheral factors regulating GHRH and SST actions. For example, key modulators of food intake and peripheral factors that signal adiposity are thought to override GHRH- and SST-mediated GH release, entraining the amount and pattern of GH release relative to calorie provision and peripheral factors associated with food intake and adiposity. Therefore, it is firstly hypothesised that the suppression of GH secretion in obesity does not occur in response to endocrine dysfunction, rather it occurs alongside progressive weight gain, and thus changes in GH feedback to promote the progressive suppression of GH relative to increased fat mass.Data within this thesis confirms the suppression of peak and total GH release in obese mice, further demonstrating a progressive suppression of GH release relative to increased body weight and adiposity in both obese and non-obese mice. Thus, it is proposed that GH deficiency in obesity may represent physiological changes in the regulation of GH release, rather than pathological changes that culminate with the development of obesity. To clarify the association between metabolic or peripheral factors that signal adiposity with spontaneous release of GH, correlation analyses revealed an inverse relationship between epididymal fat mass, leptin (secreted proportionally to adiposity) and insulin, and parameters associated with the amount of GH released in wild-type (WT) male mice. No association was observed specific in GH pulse patterning, confirming that GH patterning (represented by the interactions of hypothalamic pulse generators – GHRH and SST) remains intact regardless of feedback pressure to reduce the amount of GH release.Given compelling evidence from pharmacological studies and modified-ghrelin animal models demonstrated that acyl-ghrelin (biologically active form of ghrelin) has the capacity to induce adiposity and amplify peak GH release, and the level of acyl-ghrelin in circulation is age-dependent, it is hypothesised that acyl-ghrelin modulates the pulsatile release of growth hormone relative to adiposity and age. To address the endogenous role of acyl-ghrelin relative to the pulsatile pattern of GH secretion, spontaneous GH release in ad libitum (ad lib) fed, conscious and free-moving male mice with germline deletion of ghrelin-o-acyltransferase (goat) was assessed. These animals (goat-/-) do not have acyl-ghrelin, and thus offer a unique opportunity to define the importance of acyl-ghrelin in regulating the amount and pulsatile pattern of GH secretion. Measures were extended to include transcript expression for key hypothalamic regulators of GH release, and peripheral factors modulated relative to GH release. As anticipated, data shows a significant reduction in the amplitude of GH release in goat-/- mice compared to age-matched WT mice. Data demonstrate comparable levels of adiposity in WT and goat-/- mice. Moreover, the negative association between adiposity and GH release is conserved in both genotypes. As such, further studies were conducted to interrogate the in vivo role of acyl-ghrelin in regulating GH release relative to age.The amplitude of GH release was remarkably low in younger goat-/- mice compared to age-matched WT mice. Although it is thought that the reduction in GH release is coupled to an age-associated decline in acylated ghrelin levels, this was not observed in goat-/- mice when compared with age-matched WT mice, potentially due to low levels of GH release at a young age. Moreover, as epididymal fat weight from age-matched WT and goat-/- mice was comparable, the reduction in the amount of GH secreted in young goat-/- mice likely occurred independent of feedback that is specific to adiposity. An increase in GH pulse frequency was observed alongside irregular pulse patterning in goat-/- mice. This was characterised by an increase in the number of GH pulses and ApEn observed during extended secretory events. This overall change in GH patterning in goat-/- mice did not coincide with changes in hypothalamic Ghrh, Sst, Neuropeptide Y (Npy) or Gh secretagogue receptor (Ghsr) gene expression, or pituitary GH content, suggesting that loss of acyl-ghrelin does not alter the presence of components of central systems thought to contribute to the synthesis and release of GH. Rather, it is proposed that loss of acyl-ghrelin may alter interactions between these hypothalamic components of GH release. Of interest, despite reduced GH release, body length remained similar between goat-/- and WT mice. Furthermore, peripheral levels of insulin-like growth factor 1 (IGF-1) in goat-/- mice were positively correlated with an increase in GH pulse frequency. Thus, sustained or increased IGF-1 release in goat-/- mice may compensate for the effects of reduced peak GH release (in young adult mice) on linear growth.In summary, observations within this thesis demonstrate that, 1) weight gain and increased adiposity contributes to a progressive reduction in the amount of GH release; 2) altered secretion of leptin (as an adipostat) and insulin (a metabolic modulator of glucose metabolism) may not directly impact GH pulse patterning, rather these factors are proposed to contribute to progressive reduction in GH release relative to fat mass, and thus may contribute to the feedback regulation of peak and total GH release as obesity develops; 3) germline loss of acyl-ghrelin contributes to derangements in the amount and patterning of GH release independent of adiposity and 4) the loss of acyl-ghrelin results in a rise in the frequency and irregularity of GH pulses, which in turn may recover peripheral levels of IGF-1 to sustain linear growth. While these factors largely entrain the amount and patterning of GH release, contributing to progressive changes to meet physiological needs, derangements in GH release may be detrimental to the long-term anabolic actions of GH.

The toxicity and irritation associated with high amounts of surfactants restrict the extensive utilization of microemulsions. To address these shortcomings, employing mixed oils to enlarge microemulsion areas therefore reducing surfactant contents is a promising strategy. However, what kinds of mixed oils are more efficient in enlarging microemulsion areas still remains unclear. In this research, we found that the chain length and degree of unsaturation of oils play a key role in enlarging microemulsion areas. The combination of moderate chain saturated oil caprylic/capric triglyceride (GTCC) with long chain unsaturated oil glycerol trioleate significantly increased the microemulsion areas. Solubility of ibuprofen in the mixed oils was unexpectedly and remarkably increased (almost 300mg/mL) compared with that (around 100mg/mL) of the single oil (GTCC), which also resulted in greatly increased solubility of ibuprofen in mixed oils-containing microemulsions. By optimizing the mixed oil formulation, the absolute amount of surfactant in drug-loaded microemulsions was reduced but increased drug oral bioavailability in rats was maintained. It could be concluded that the combined use of moderate chain oils and long chain unsaturated oils could not only acquire enlarged microemulsion areas but also enhanced drug solubility, therefore doubly reducing surfactant amount, which is extremely beneficial for developing safe microemulsions.

So far, the components responsible for the neuroprotective effects of Calculus bovis are unclear. Cholesterol, one of the major components in Calculus bovis, is easily oxidized into oxysterols, which possess direct or indirect neuroprotective effects proved by our and others' previous studies. Therefore, a liquid chromatography with mass spectrometry method coupled with ultrasonic extraction and solid-phase extraction was developed for the determination of neuroprotective oxysterols in Calculus bovis, human gallstones, and traditional Chinese medicine preparations. Chromatographic separation was achieved on a C18 column with isocratic elution at a flow rate of 1 mL/min. The established method showed good linearity (R(2) > 0.998), sensitivity with low limits of detection (0.06-0.39 μg/g), acceptable precisions (relative standard deviations ≤ 7.4%), stability (relative standard deviations ≤ 5.9%), and satisfactory accuracy (92.4-102.9%) for all analytes identified by different retention times, which could be applied for the determination of oxysterols. Five kinds of oxysterols proved to function as neuroprotectants were detected at different concentrations. Among them, 7β-hydroxycholesterol and cholestane-3β,5α,6β-triol were rather abundant in the samples. It could be concluded that the potential neuroprotective components in Calculus bovis may be these oxysterols.

This research aims to synthesize lipophilic berberine derivatives and evaluate their antiglioma effects on C6 and U87 cells. The introduction of substituents with various carbon chain lengths on C-13- or C-9-O-position of the berberine scaffold led to the discovery of several potent inhibitors against glioblastoma cells. Derivatives substituted with the carbon chains of moderate length (twelve carbons) displayed improved lipophilicity and the strongest inhibitory effects. Several compounds presented dose-dependent repression against proliferation (IC50, 1.12–6.12 μM) and blocked migration and invasion by over 60% at lower dose levels. Furthermore, preliminary research about the underlying mechanism for the enhanced antiglioma ability indicated that these analogues preferentially localized into mitochondria, inducing the up-regulation of ROS production. Overall, these compounds represent promising candidates to combat glioblastoma and highlight new insight into the antiglioma therapy through interaction with mitochondria.

Sijia Zheng,1,* Yanqi Xie,1,* Yuan Li,1 Ling Li,1 Ning Tian,1 Wenbo Zhu,2 Guangmei Yan,2 Chuanbin Wu,1 Haiyan Hu1 1School of Pharmaceutical Sciences, 2Department of Pharmacology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, People's Republic of China *These authors contributed equally to this workAbstract: The objective of this research was to develop and evaluate high drug-loading ligand-modified nanomicelles to deliver a steroidal compound to the brain. YC1 (5α-cholestane-24-methylene-3β, 5α, 6β, 19-tetraol), with poor solubility and limited access to the brain, for the first time, has been proved to be an effective neuroprotective steroid by our previous studies. Based on the principle of ‘like dissolves like’, cholesterol, which shares the same steroidal parent nucleus with YC1, was selected to react with sodium alginate, producing amphiphilic sodium alginate–cholesterol derivatives (SACDs). To increase the grafting ratio and drug loading, cholesterol was converted to cholesteryl chloroformate, for the first time, before reacting with sodium alginate. Further, lactoferrin was conjugated on SACDs to provide lactoferrin-SACDs (Lf-SACD), which was established by immune electron microscopy (IEM) and self-assembled into brain-targeting nanomicelles. These nanomicelles were negatively charged and spherical in nature, with an average size of

In an effort to screen natural compounds and their derivatives as novel agents for cancer treatment, we developed two semisynthetic nature compounds, A14 and A52, which have potencies in an in vitro cell proliferation studies against LS174T and PC-3 cells in the mid-nanomolar range. A14 inhibited more than 90% of in vitro cancer cell proliferation around 300 nM and also inhibited in vivo PC-3 prostate cancer xenografts in nude mice. Cell morphology studies and comparisons with known antineoplastic agents suggested that A14 disrupted microtubule organization. In vitro tubulin polymerization assay suggested that A14 inhibited this process. Based on molecular docking, A14 fits very well in the colchicine-binding site at the interface between the alpha-tubulin and beta-tubulin. Further analyses suggest that A14 sensitivity may correlate with CDK/cyclin activity. A14 inhibited PC-3 prostate cancer xenografts in nude mice. In addition to solid tumors, A14 and its analog A52 strongly inhibited a panel of human leukemia cancer cell lines. Furthermore, these compounds inhibited Myc-induced T cell acute lymphoblastic leukemia (T-ALL) in a zebrafish model. Although many tubulin inhibitors were available, there is no compound that targeting the colchicine-binding site has been approved by FDA for cancer treatment. This new family of tubulin polymerization inhibitors could be further developed for the treatment of both solid and liquid tumors. Citation Format: Yanqi Xie, Liliia Kril, Wen Zhang, Mykhaylo Frasinyuk, Svitlana Bondarenko, Elizabeth Hausman, Zachary Martin, Przemyslaw Wyrebek, Tianxin Yu, Changguo Zhan, Vitaliy Sviripa, Jessica Blackburn, David Watt, Chunming Liu. Novel tubulin polymerization inhibitors repress tumor xenografts in nude mice and leukemia in zebrafish [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2674.

So far, the components responsible for the neuroprotective effects of Calculus bovis are unclear. Cholesterol, one of the major components in Calculus bovis, is easily oxidized into oxysterols, which possess direct or indirect neuroprotective effects proved by our and others' previous studies. Therefore, a liquid chromatography with mass spectrometry method coupled with ultrasonic extraction and solid-phase extraction was developed for the determination of neuroprotective oxysterols in Calculus bovis, human gallstones, and traditional Chinese medicine preparations. Chromatographic separation was achieved on a C18 column with isocratic elution at a flow rate of 1 mL/min. The established method showed good linearity (R(2)0.998), sensitivity with low limits of detection (0.06-0.39 μg/g), acceptable precisions (relative standard deviations ≤ 7.4%), stability (relative standard deviations ≤ 5.9%), and satisfactory accuracy (92.4-102.9%) for all analytes identified by different retention times, which could be applied for the determination of oxysterols. Five kinds of oxysterols proved to function as neuroprotectants were detected at different concentrations. Among them, 7β-hydroxycholesterol and cholestane-3β,5α,6β-triol were rather abundant in the samples. It could be concluded that the potential neuroprotective components in Calculus bovis may be these oxysterols.

In this paper, we introduce a Web data mining solution to e-learning system to discover hidden patterns strategies from their learners and Web data, describe a personalized recommender system that uses web mining techniques for recommending a student which (next) links to visit within an adaptable e-learning system, propose a new framework based on data mining technology for building a Web-page recommender system, and demonstrate how data mining technology can be effectively applied in an e-learning environment.