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Most genetic variants for colorectal cancer (CRC) identified in genome-wide association studies (GWAS) are located in intergenic regions, implying pathogenic dysregulations of gene expression. However, comprehensive assessments of target genes in CRC remain to be explored. We conducted a multi-omics analysis using transcriptome and/or DNA methylation data from the Genotype-Tissue Expression, The Cancer Genome Atlas and the Colonomics projects. We identified 116 putative target genes for 45 GWAS-identified variants. Using summary-data-based Mendelian randomization approach (SMR), we demonstrated that the CRC susceptibility for 29 out of the 45 CRC variants may be mediated by cis-effects on gene regulation. At a cutoff of the Bonferroni-corrected PSMR

The current procedure thoroughly explains how to reprogram induced pluripotent stem cells (iPSCs) from the patient's peripheral blood mononuclear cells (PBMCs), a less invasive source; e.g., somatic cells. We describe how to isolate PBMCs and reprogram them into iPSCs by electroporation. Furthermore, we provide an alternative approach to generating iPSC using Geltrex or Matrigel matrix to replace MEF-feeder. The challenge of this process is the relatively lower cell survival rates of PBMCs due to the damage of electroporation. For complete details on the use and execution of this protocol, please refer to Hu et al. (2021).

Colorectal cancer (CRC) is a prevalent and fatal cancer. Oral administration provided the potential for in situ treatment of the colorectal cancer. However, drugs couldn't be well-absorbed mainly due to its degradation in the gastric area and poor intestinal permeability. In this study, we synthesized deoxycholic acid and hydroxybutyl decorated chitosan nanoparticles (DAHBC NPs) as oral curcumin (CUR) delivery system for colorectal cancer treatment. DAHBC with lower critical solution temperature (LCST) below 37 °C (27-33 °C) was obtained. DAHBC NPs were correspondingly stable in simulated gastric conditions (pH 1.2, 37 °C), due to the offset of size change between pH-responsive expansion and thermo-responsive shrinkage. In simulated intestinal tract (pH 7.0-7.4, 37 °C), DAHBC NPs exhibited burst release of CUR owing to the onefold effect of thermo-responsive shrinkage. DAHBC27 NPs showed the minimum CUR leakage (~10%) in simulated gastric conditions, because a furthest temperature-sensitive shrinkage caused by the lowest LCST offset the expansion in acid environment. DAHBC27 NPs induced ~10-fold increased (P 0.05) CUR absorption by paracellular transport pathway, compared to the free CUR. Thus, DAHBC NPs stabilized in the gastric environment may be a promising oral drugs delivery system for effective in situ colorectal cancer therapy.

BACKGROUND AND AIMS: Susceptibility genes and the underlying mechanisms for the majority of risk loci identified by genome-wide association studies (GWAS) for colorectal cancer (CRC) risk remain largely unknown. We conducted a transcriptome-wide association study (TWAS) to identify putative susceptibility genes. METHODS: Gene-expression prediction models were built using transcriptome and genetic data from the 284 normal transverse colon tissues of European descendants from the Genotype-Tissue Expression (GTEx), and model performance was evaluated using data from The Cancer Genome Atlas (TCGA, n = 355). We applied the gene-expression prediction models and GWAS data to evaluate associations of genetically predicted gene-expression with CRC risk in 58,131 CRC cases and 67,347 controls of European ancestry. Dual-luciferase reporter assays and knockdown experiments in CRC cells and tumor xenografts were conducted. RESULTS: We identified 25 genes associated with CRC risk at a Bonferroni-corrected threshold of P < 9.1 × 10(−6), including genes in four novel loci, PYGL (14q22.1), RPL28 (19q13.42), CAPN12 (19q13.2), MYH7B (20q11.22), and MAP1L3CA (20q11.22). In nine known GWAS-identified loci, we uncovered nine genes that have not been previously reported, whereas four genes remained statistically significant after adjusting for the lead risk variant of the locus. Through colocalization analysis in GWAS loci, we additionally identified 12 putative susceptibility genes that were supported by TWAS analysis at P < 0.01. We showed that risk allele of the lead risk variant rs1741640 affected the promoter activity of CABLES2. Knockdown experiments confirmed that CABLES2 plays a vital role in colorectal carcinogenesis. CONCLUSION: Our study reveals new putative susceptibility genes and provides new insight into the biological mechanisms underlying CRC development.

e15125 Background: Upper tract urothelial carcinoma (UTUC) is histologically similar to bladder urothelial carcinoma (BUC), yet several clinical, biological and molecular features are unique to UTUC. UTUC may have unique genomic factors compared to BUC, while major knowledge gaps remain in our understanding of the biology and genomic landscape of UTUC. Methods: We retrospectively sequenced tumor samples and matched germline DNA using targeted next-generation sequencing methods and analyzed at least 642 cancer-associated genes. The cohort included 35 UTUC patients and 36 BUC patients. Results: Among all cases, KMT2D and TP53 were the most common mutations (account for 66% and 75% in UTUC and BUC, respectively). The most frequently mutated genes in UTUC were KMT2D (37%), TP53 (29%), ELF3 (26%), FGFR3 (23%), ASXL1 (20%), DKN2A (20%), DKN2B (17%), REBBP (17%), MT3B (17%), GNAS (17%) and PIK3CA (17%). The most frequently mutated genes in BUC were KMT2D (36%), TP53 (39%), MUC17 (14%), ARID1A (22%), GNAS (19%), RB1 (19%), BCL6 (19%), E2F3 (19%), EIF4A2 (19%) and FGF3 (19%). However significant differences in the prevalence of mutations in individual genes were observed. FGFR3 (23% vs. 8%) and ELF3 (26% vs. 11%) were more frequently altered, whereas RB1 (6% vs. 19%), E2F3 (3% vs. 19%) and BCL6 (3% vs. 19%) were less frequently altered in UTUC (P < 0.05). FGFR3 mutations have been shown to be oncogenic and RB1 inactivation has been associated with genomic instability in urothelial cancer in previous researches, which represent potential therapeutic targets. Conclusions: Â UTUC and UCB exhibit significant differences in the prevalence of genomic landscape and carcinogenesis. A comprehensive understanding of the biology of UTUC and UCB is needed to identify new drug targets in order to improve clinical outcomes.

The low-rank representation (LRR) was presented recently and demonstrated its effectiveness for robust subspace segmentation. This paper presents a discriminative projection method based on Low-rank affinity matrix (LRA-DP) for robust feature extraction. The affinity matrix is designed to better preserve the underlying low-rank structure of data representation revealed by LRR. The experiments on the Yale, Extended Yale B, AR face image databases and the PolyU palmprint database showed LRA-DP is always better than or comparable to other state-of-the-art methods, which means underlying low-rank structure of data representation preserved by LRA-DP is helpful for classification problem.

Herb mixtures are widely used for treatment of the menopausal syndrome long before the hormonal therapy. However, there is insufficient data for herb remedies in treating menopausal syndromes. Here we aim to investigate the effect of ZhenRongDan (ZRD) in balancing female hormones, regulating expression of estrogen receptors (ERs), and preventing organ atrophy in menopausal rats. Rats that underwent bilateral ovariectomy were used in the experiments; the effects of ZRD on serum follicle-stimulating hormone (FSH), luteinizing hormone (LH), prolactin (PRL), and estradiol (E2) levels were observed. Histology of vagina and ERs expression in vagina, uterus, and adrenal gland were also examined. ELISAs were used to analyze the changes of FSH, LH, PRL, and E2 in serum, and the morphological changes of the cervical epithelium cells were observed by Hematoxylin & Eosin (H&E) staining. Immunohistochemistry and western blot were applied to detect estrogen receptors subtypes alpha (ERα) and beta (ERβ) expression in vagina, uterus, and adrenal gland. We found that ZRD could significantly reduce the weight of the adrenal gland and increase the weight of the uterus. It could decrease the release of FSH and LH as well as increasing E2 and PRL levels. Furthermore, ZRD could improve the number of cervical vaginal epithelial cells and increase the thickness of the vaginal wall. And the altered expressions of ERαand ERβare also restored by ZRD. ZRD could obviously relieve the endocrine disorders, modulate organ atrophy, and restore ERαand ERβexpression in the ovariectomized rat model.

The objective of this study was to isolate fucoxanthin from Sargassum thunbergii and develop microcapsules with palm stearin as the solid lipid core for stability and efficient oral delivery of fucoxanthin. The microcapsules had smooth surfaces with the volume weighted mean diameter (d4.3) of 19.19 μm. Encapsulation efficiency and loading capacity of microcapsules with fucoxanthin were 98.3% and 0.04%, respectively. Moreover, the fucoxanthin in microcapsules presented higher stability than free fucoxanthin against light, humidity and temperature. Especially, the retention rates of fucoxanthin encapsulated in microcapsules reached 97.20% at 4°C, 92.60% at 25°C, 92.32% with the relative humidity of 33% and 92.60% in the dark. The cumulative amount of fucoxanthin released from microcapsules was 22.92% in simulated gastric fluid (SGF) and 56.55% in simulated intestinal fluid (SIF).

Acute kidney injury (AKI) is common in hospital patients. Delayed diagnosis and treatment of AKI due to the lack of efficient early diagnosis is an important cause of its high mortality. While fluorescence imaging seems promising to non-intrusively interrogate AKI-related biomarkers, the low kidney contrast of many fluorophores conferred by their relatively low abundance of distribution in the kidney limits their application for AKI detection. Herein, we discovered a near-infrared fluorophore with inherent kidney-targeting ability. Based on this fluorophore, a fluorogenic probe (KNP-1) was developed by targeting peroxynitrite (ONOO−), which is upregulated at the early onset of AKI. KNP-1 exhibits desirable kidney distribution after intravenous administration and is fluorescent only after activation by ONOO−. These properties lead to excellent kidney contrast imaging results. KNP-1 is capable of detecting both nephrotoxin-induced and ischemia-reperfusion injury-induced AKI in live mice. Temporally resolved imaging of AKI-disease model mice with KNP-1 suggests a gradual increase in renal ONOO− levels with disease progression. Notably, the upregulation of ONOO− can be observed at least 24 h earlier than the clinically popular sCr and BUN methods. Blocking ONOO− generation also proves beneficial. These results highlight the applicability of this inherently tissue targeting-based strategy for designing probes with desirable imaging contrast; potentiate ONOO− as a biomarker and target for AKI early diagnosis and medical intervention; and imply the clinical relevance of KNP-1 for AKI early detection.

e17515 Background: Philadelphia Prostate Cancer Consensus Conference 2017 developed an expert consensus-driven working framework for comprehensive genetic evaluation of inherited PCA. However, the consensus was mainly based on data from Caucasian populations. Genetic differences in the PC molecular landscape between Asian and Caucasian men have been established. Whether Caucasian-based genetic information can be used to guide clinical practice in Chinese population needs further evidence. Methods: 1123 patients with confirmed prostate cancer admitted from March 2018 to August 2019 from 18 Chinese centers were included in this retrospective study. We sequenced the association of 50 gene with prostate cancer risk, clinical treatment implications and predisposition, focusing on DNA repair gene and genetic susceptibility genes pathogenic mutations in Philadelphia Prostate Cancer Consensus Conference. Only protein-truncating alterations (nonsense/stop-gains, frameshift insertions and deletions, and donor and acceptor splice-site mutations) were coded as pathogenic or likely pathogenic protein-truncating alterations. Results: Among all cases, DNA repair gene pathogenic mutations were found in 133 men (11.84%), including BRCA2 (62 [5.5%]), ATM (15 [1.3%]), PALB2 (11 [0.97%]), GEN1 (7 [0.62%]), BRCA1 (6 [0.53%]), MLH3 (4 [0.35%]), MUTYH (4 [0.35%]), PMS1 (4 [0.35%]), RAD50 (4 [0.35%]), ATR (3 [0.27%]), CHEK2 (3 [0.27%]), FANCD2 (3 [0.27%]) etc. Similar germline DRG mutation frequencies were observed between our cohort and Caucasian men. Noticeably, G84E mutation in HOXB13, a rare but recurrent missense mutation associated with a significantly increased risk of hereditary prostate cancer was not found in our cohort of patients. Furthermore, the frequency of HSD3B1 (1245C) allele in the Chinese patients is much lower than that in the Caucasian populations (11.7% vs 63%). Conclusions: Our study retrospectively analyzed germline mutation profile of prostate cancer in Chinese population. The frequency of HSD3B1 (1245C) allele varies across ethnic populations in spite of the similar DNA repair gene mutations rates. Considering the gene list should be tested based on clinical/familial scenarios from the Philadelphia Prostate Cancer Consensus Conference, a prevalence of certain mutations, G84E mutation in HOXB13 was unnecessary for genetic testing in Chinese population.

Radiation-induced lung fibrosis (RILF) is a long-term adverse effect of curative radiotherapy. The accumulation of myofibroblasts in fibroblastic foci is a pivotal feature of RILF. In the study, we found the inhibitory effect of grape seed proanthocyanidins (GSPs) on irradiation-induced differentiation of human fetal lung fibroblasts (HFL1). To explore the mechanism by which GSPs inhibit fibroblast differentiation, we measured the reactive oxygen species (ROS) levels, mitochondrial function, mitochondrial dynamics, glycolysis and the signaling molecules involved in fibroblast transdifferentiation. GSPs significantly reduced the production of cellular and mitochondrial ROS after radiation. The increases in mitochondrial respiration, proton leak, mitochondrial ATP production, lactate release and glucose consumption that occurred in response to irradiation were ameliorated by GSPs. Furthermore, GSPs increased the activity of complex I and improved the mitochondrial dynamics, which were disturbed by irradiation. In addition, the elevation of phosphorylation of p38MAPK and Akt, and Nox4 expression induced by irradiation were attenuated by GSPs. Blocking Nox4 attenuated irradiation-mediated fibroblast differentiation. Taken together, these results indicate that GSPs have the ability to inhibit irradiation-induced fibroblast-to-myofibroblast differentiation by ameliorating mitochondrial dynamics and mitochondrial complex I activity, regulating mitochondrial ROS production, ATP production, lactate release, glucose consumption and thereby inhibiting p38MAPK-Akt-Nox4 pathway.

Ferulate 5-hydroxylase (F5H), a cytochrome P450 monooxygenase, has been indicated to function in the monolignol biosynthesis of sinapyl alcohol, namely the precursor of syringyl lignin during phenylpropanoid metabolism. In the present study, a full-length cDNA clone encoding F5H, designated as TbF5H1, was cloned from water chestnuts (Trapa bicornis Osbeck.) using RT-PCR and RACE. Sequence analysis of TbF5H1 protein revealed the presence of highly conserved motifs shard by P450 s, such as heme binding cysteine, oxygen binding I-helix, and E-R-R triade. Subsequently, TbF5H1 was expressed in bacterial system as a recombinant protein, which was used to ascertain kinetic parameters of the protein with the Km and Vmax values of 2.40 μM and 3.35 pkat mg−1, respectively. Together with high identity with other plant F5Hs based amino acid sequences, this result suggested that TbF5H1 had a biological function similar to its homologues in S lignin synthesis. Western blotting analysis using anti-TbF5H1 serum towards total proteins extracted from roots, stems, leaves, and flowers, detected a protein with a molecular weight of about 57 kDa. Quantitative real-time PCR analysis revealed the transcript level of TbF5H1 was higher in leaves than in other tested tissues. Interestingly, TbF5H1 was differentially expressed in response to various stresses, including salt (NaCl), hydrogen peroxide (H2O2) and heavy metal (lead), as well as elicitation by methyl ester jasmonate (MeJA). The expression of TbF5H1 was not significantly altered under NaCl or lead stress conditions, whereas a significant increase in TbF5H1 expression was detected after elicitation of H2O2 and MeJA, respectively. Moreover, there existed a fine coordination between expression of TbF5H1 and contents of certain nonenzymatic antioxidants. These results indicated that integrated with some redox components, TbF5H1 gene should contribute to the acclimation of T. bicornis to oxidative stress, which is modulated by MeJA.

In this work, the composite NiNdxFe2−xO4/NanoG was prepared by coprecipitation reaction of Nd3+ substituted nickel ferrite with graphite nanosheet (NanoG). The structures of NiNdxFe2−xO4 (x=0.03, 0.06, 0.09) and NiNdxFe2−xO4/NanoG were characterized by scanning electron microscopy, energy dispersive spectroscopy, X-ray diffraction and laser particle size appliance. Results show that when x=0.06, the particle size of NiNd0.06Fe1.94O4 is in the range of 60–70 nm and can be chemically precipitated on the surface of NanoG. The saturation magnetization of NiNd0.06Fe1.94O4 is 41.79 emu/g while it is 26.24 emu/g for NiNd0.06Fe1.94O4/NanoG. The conductivity of NiNd0.06Fe1.94O4/NanoG is in the semiconductor sphere. Measurement of electromagnetic parameters shows that the minimum reflection loss of NiNd0.06Fe1.94O4 is −13.15 dB at 11.88 GHz. When the mass ratio of NiNd0.06Fe1.94O4 to NanoG reaches 4:1, the minimum reflection loss of NiNd0.06Fe1.94O4/NanoG is −17.03 dB at 9.29 GHz.

Here we described nano-polyplexes (NPs) made of oleoyl-carboxymethy-chitosan (OCMCS)/hyaluronic acid (HA) as novel potential carriers for oral gene vaccines delivery. Aerolysin gene (aerA) of Aeromonas hydrophila as microbial antigen was efficiently loaded to form OCMCS-HA/aerA (OHA) NPs. OHA NPs performed the optimal parameters, i.e. smallest (154.5±9.4nm), positive charged (+7.9±0.5mV) and monodispersed system with the N/P ratio of 5 and OCMCS/HA weight ratio of 4. Upon the introduction of HA, OHA NPs was beneficial for the DNA release in intestinal environments in comparison to OA NPs. The mean fluorescence intensity detected in Caco-2 cells incubated with OHA NPs was about 2.5-fold higher than that of OA NPs; however, it decreased significantly in the presence of excess free HA. The OHA NPs and OA NPs decreased the transepithelial electric resistance (TEER) of Caco-2 monolayers obviously and induced increasing the apparent permeability coefficient (Papp) of DNA by 5.45-6.09 folds compared with free DNA. Significantly higher (P

In order to design a high-performance electron gun, a new design concept was adapted to reduce the spherical aberration and coma of the electron optical system. In the electron gun we used a new asymmetric beam-forming region (BFR) and asymmetric main lens. This gun has been used for 40 cm 0.28 mm-pitch full square color display tubes. At cathode current of 0.2 microampere, the spot size on the screen center was measured 0.65 mm (10% of the maximum current distribution), and at the cathode current of 0.2 - 0.5 microampere, the focus voltage difference (focus trial characteristic) is less than 150 V. When the horizontal- scanning frequency reached 57 kHz, the tubes using the electron gun can display 1024 X 768 dots, and the corner- center focus performance is good. So it can meet the requirements of 40 cm 0.28 mm CDT of the market. In this design we used computer simulation software called Electron Beam Simulation to simulate the BFR and the main lens, through it we can adjust the lens structure, predict the performance and make it better. A good coincidence is obtained between the calculations and experiment results.© (1998) COPYRIGHT SPIE--The International Society for Optical Engineering. Downloading of the abstract is permitted for personal use only.