Molecular Mechanisms and Tumor Biological Aspects of 5-Fluorouracil Resistance in HCT116 Human Colorectal Cancer Cells

5-Fluorouracil (5-FU) is a cornerstone drug used in the treatment of colorectal cancer (CRC). However, the development of resistance to 5-FU and its analogs remain an unsolved problem in CRC treatment. In this study, we investigated the molecular mechanisms and tumor biological aspects of 5-FU resistance in CRC HCT116 cells. We established an acquired 5-FU-resistant cell line, HCT116R^F10. HCT116R^F10 cells were cross-resistant to the 5-FU analog, fluorodeoxyuridine. In contrast, HCT116R^F10 cells were collaterally sensitive to SN-38 and CDDP compared with the parental HCT16 cells. Whole-exome sequencing revealed that a cluster of genes associated with the 5-FU metabolic pathway were not significantly mutated in HCT116 or HCT116R^F10 cells. Interestingly, HCT116R^F10 cells were regulated by the function of thymidylate synthase (TS), a 5-FU active metabolite 5-fluorodeoxyuridine monophosphate (FdUMP) inhibiting enzyme. Half of the TS was in an active form, whereas the other half was in an inactive form. This finding indicates that 5-FU-resistant cells exhibited increased TS expression, and the TS enzyme is used to trap FdUMP, resulting in resistance to 5-FU and its analogs.     

MiR-10a in Pancreatic Juice as a Biomarker for Invasive Intraductal Papillary Mucinous Neoplasm by miRNA Sequencing

New biomarkers are needed to further stratify the risk of malignancy in intraductal papillary mucinous neoplasm (IPMN). Although microRNAs (miRNAs) are expected to be stable biomarkers, they can vary owing to a lack of definite internal controls. To identify universal biomarkers for invasive IPMN, we performed miRNA sequencing using tumor-normal paired samples. A total of 19 resected tissues and 13 pancreatic juice samples from 32 IPMN patients were analyzed for miRNA expression by next-generation sequencing with a two-step normalization of miRNA sequence data. The miRNAs involved in IPMN associated with invasive carcinoma were identified from this tissue analysis and further verified with the pancreatic juice samples. From the tumor-normal paired tissue analysis of the expression levels of 2792 miRNAs, 20 upregulated and 17 downregulated miRNAs were identified. In IPMN associated with invasive carcinoma (INV), miR-10a-5p and miR-221-3p were upregulated and miR-148a-3p was downregulated when compared with noninvasive IPMN. When these findings were further validated with pancreatic juice samples, miR-10a-5p was found to be elevated in INV (p = 0.002). Therefore, three differentially expressed miRNAs were identified in tissues with INV, and the expression of miR-10a-5p was also elevated in pancreatic juice samples with INV. MiR-10a-5p is a promising additional biomarker for invasive IPMN.     

Identification of SYS1 as a Host Factor Required for Shiga Toxin-Mediated Cytotoxicity in Vero Cells

Shiga toxin (STx) or Vero toxin is a virulence factor produced by enterohemorrhagic Escherichia coli. The toxin binds to the glycosphingolipid globotriaosylceramide (Gb3) for its entry, and causes cell death by inhibiting ribosome function. Previously, we performed a loss-of-function screen in HeLa cells using a human CRISPR knockout (KO) library and identified various host genes required for STx-induced cell death. To determine whether this library targeted to the human genome is applicable to non-human primate cells and to identify previously unrecognized factors crucial for STx-induced cell death, we herein performed a similar screen in the African green monkey kidney-derived Vero C1008 subline. Many genes relevant to metabolic enzymes and membrane trafficking were enriched, although the number of enriched genes was less than that obtained in the screening for HeLa cells. Of note, several genes that had not been enriched in the previous screening were enriched: one of these genes was SYS1, which encodes a multi-spanning membrane protein in the Golgi apparatus. In SYS1 KO Vero cells, expression of Gb3 and sphingomyelin was decreased, while that of glucosylceramide and lactosylceramide was increased. In addition, loss of SYS1 inhibited the biosynthesis of protein glycans, deformed the Golgi apparatus, and perturbed the localization of trans-Golgi network protein (TGN) 46. These results indicate that the human CRISPR KO library is applicable to Vero cell lines, and SYS1 has a widespread effect on glycan biosynthesis via regulation of intra-Golgi and endosome–TGN retrograde transports.     

Genetic Susceptibility to Periodontal Disease in Down Syndrome: A Case-Control Study

Severe periodontitis is prevalent in Down syndrome (DS). This study aimed to identify genetic variations associated with periodontitis in individuals with DS. The study group was distributed into DS patients with periodontitis (n = 50) and DS patients with healthy periodontium (n = 36). All samples were genotyped with the “Axiom Spanish Biobank” array, which contains 757,836 markers. An association analysis at the individual marker level using logistic regression, as well as at the gene level applying the sequence kernel association test (SKAT) was performed. The most significant genes were included in a pathway analysis using the free DAVID software. C12orf74 (rs4315121, p = 9.85 × 10⁻⁵, OR = 8.84), LOC101930064 (rs4814890, p = 9.61 × 10⁻⁵, OR = 0.13), KBTBD12 (rs1549874, p = 8.27 × 10⁻⁵, OR = 0.08), PIWIL1 (rs11060842, p = 7.82 × 10⁻⁵, OR = 9.05) and C16orf82 (rs62030877, p = 8.92 × 10⁻⁵, OR = 0.14) showed a higher probability in the individual analysis. The analysis at the gene level highlighted PIWIL, MIR9-2, LHCGR, TPR and BCR. At the signaling pathway level, PI3K-Akt, long-term depression and FoxO achieved nominal significance (p = 1.3 × 10⁻², p = 5.1 × 10⁻³, p = 1.2 × 10⁻², respectively). In summary, various metabolic pathways are involved in the pathogenesis of periodontitis in DS, including PI3K-Akt, which regulates cell proliferation and inflammatory response.     

SBR法处理柠檬酸废水pH变化规律研究

探索了用SBR法处理柠檬酸废水过程中pH的变化规律。研究了柠檬酸废水处理过程中pH变化与COD处理效率的关系，研究目的在于利用易于检测的水质参数为实现SBR处理过程的自动监测和调控作基础试验研究。     

SBR反应器中反硝化条件下去除苯酚工艺

基于异养反硝化原理,在序批式间歇反应器(SBR)中对反硝化同时降解苯酚的菌种进行了149 d的驯化,驯化通过逐步提高进水中苯酚和NO3--N的浓度进行.驯化结束后,进水苯酚质量浓度达到360 mg/L,葡萄糖质量浓度达到100 mg/L,NO3--N质量浓度达到240 mg/L,水力停留时间6 h,苯酚和NO3--N去除率均大于98%.反应器运行结果表明:进水苯酚质量浓度低于720 mg/L时,SBR反应器运行稳定;高浓度NO2--N(＞60 mg/L)可以严重影响微生物对苯酚以及NO3--N的去除能力,同时反应器中20.5%～23.5%的COD可被用于微生物的细胞合成.     

UASB-SBR工艺处理城市生活污水的研究

研究了UASB工艺处理城市生活污水的可行性，通过UASB与SBR组合工艺的运行实验，得出在水力停留时间3h时，COD去除率为61．67％-82．32％，出水COD达到国家污水排放二级标准。在氮磷去除效果不佳的情况下，采用SBR工艺处理后，其出水COD、SS、氮、磷等指标均达到国家污水一级排放标准。     

Modeling and solving mixed-model assembly line balancing problem with setups. Part I: A mixed integer linear programming model

This paper is the first one of the two papers entitled “modeling and solving mixed-model assembly line balancing problem with setups”, which has the aim of developing the mathematical programming formulation of the problem and solving it with a hybrid meta-heuristic approach. In this current part, a mixed-integer linear mathematical programming (MILP) model for mixed-model assembly line balancing problem with setups is developed. The proposed MILP model considers some particular features of the real world problems such as parallel workstations, zoning constraints, and sequence dependent setup times between tasks, which is an actual framework in assembly line balancing problems. The main endeavor of Part-I is to formulate the sequence dependent setup times between tasks in type-I mixed-model assembly line balancing problem. The proposed model considers the setups between the tasks of the same model and the setups because of the model switches in any workstation. The capability of our MILP is tested through a set of computational experiments. Part-II tackles the problem with a multiple colony hybrid bees algorithm. A set of computational experiments is also carried out for the proposed approach in Part-II.