Abstract Objectives: The role of Epoxide Hydrolase-4 (EPHX4), a member of epoxide hydrolase family, has not been investigated in cancer. The purpose of this article is to explore the application value of EPHX4 in laryngeal cancer and its relationship with immune infiltration. Methods: We observed that EPHX4 expression and its survival assays in laryngeal cancer specimens based on The Cancer Genome Atlas (TCGA) cohorts. We also analyzed the correlation between immune cell infiltration levels and EPHX4 gene copy number in laryngeal cancer. Finally, we conducted in vitro assay to evaluate the functions of EPHX4 in laryngeal cancer cell line. Results: EPHX4 is highly expressed in laryngeal cancer specimens and has a poor prognosis. EPHX4 related immune cell analysis showed that it participated in NK Natural killer cell mediated cytotoxicity. Finally, Cell experiments indicate that EPHX4 could promote laryngeal cancer cell line proliferation, colony formation and invasion. Conclusions: Our research results suggest that EPHX4 may be a potential immunotherapy target for laryngeal cancer. The nominated immune signature is a helpful and promising prognostic indicator in laryngeal cancer. Levels of Evidence: Level 3. Objectives Hydrolase4 Hydrolase 4 Hydrolase- EPHX4, EPHX , (EPHX4) family Methods TCGA (TCGA cohorts Finally Results prognosis cytotoxicity proliferation invasion Conclusions Evidence 3 (EPHX4 (EPHX

ABSTRACT Returning agricultural waste/by-products to the field is an effective management measure to improve soil fertility and maintain crop productivity in agroecosystems. However, we still have limited understanding of the complex response of sisal (Agave sisalana Perrine) growth, soil nutrients and microbial communities to the long-term sisal residue return to the ecological system. This study aimed to investigate the comprehensive effects of returning sisal residue to the field on sisal yield, soil nutrient, and microbial community. Based on a four-year field experiment, the results showed that returning sisal residue to the field increased the annual yield of sisal by 30.77–33.36%. Compared with control group, sisal residue inputs significantly improved soil total nitrogen, available phosphorus, and available potassium by 46.59, 3.02, and 1.21%, respectively. Under sisal residue input condition, the activities of soil catalase, urease, acid phosphatase, and sucrase also significantly increased by 2.01, 17.20, 17.40 and 16.60%, respectively. A higher ratio of bacteria/fungi was observed in the soil amended with sisal residue. The microbial diversity analysis showed that the α-diversity of the bacterial community increased, while the α-diversity of the fungal community decreased with the sisal residue treatment. However, sisal residue return had a higher impact on the α-diversity of fungi. This study provides evidence that returning sisal residue to the field affects sisal productivity by regulating nutrient cycling and soil microbial community. Moreover, the study suggests that microbial α-diversity has significant influence on sisal yield. waste/byproducts wastebyproducts waste/by products waste agroecosystems However Agave Perrine growth longterm long term system fouryear four year experiment 30773336 30 77 33 36 30.77–33.36% group nitrogen phosphorus 4659 46 59 46.59 302 3 02 3.02 121 1 21 1.21% respectively condition catalase urease phosphatase 201 2 01 2.01 1720 17 20 17.20 1740 40 17.4 1660 16 60 16.60% bacteriafungi bacteria fungi αdiversity α treatment Moreover byproducts wasteby 3077333 7 30.77–33.36 465 4 5 46.5 0 3.0 12 1.21 2.0 172 17.2 174 17. 166 6 16.60 307733 30.77–33.3 46. 3. 1.2 2. 16.6 30773 30.77–33. 1. 16. 3077 30.77–33 307 30.77–3 30.77– 30.77 30.7 30.

Abstract The purple glume-tip is an essential morphological marker for selective rice breeding, aiding in assisted selection and variety purification. However, the inheritance of purple glume-tip in japonica rice landrace Donglan Black Rice (DBR) has not yet been explored deeply. The F2 and F4 populations were constructed from crossing between Huazhan with colorless glume-tip and DBR to identify the associated genomic region(s). Genetic analysis displayed two highly comparable and perplexing phenotypes in the purple glume-tip of rice. Two significant genes with recessive epistasis predominantly regulated the two phenotypes. The two target gene loci were located in the intervals of 5315163-5316875 bp on chr6 and 27915598-27939357 bp on chr4, respectively, where reported genes associated with the purple color trait in rice, Os06g0205100 and Os04g0557500, are present. The two genes may be potential target genes. However, the role of Os04g0557500 in the glume-tip coloration remains unreported. glumetip glume tip breeding purification However (DBR deeply F regions. regions region s . region(s) 53151635316875 5315163 5316875 5315163-531687 chr 2791559827939357 27915598 27939357 27915598-2793935 chr4 respectively Osg Os g Os06g020510 present Os04g055750 unreported region(s 5315163531687 531516 531687 5315163-53168 279155982793935 2791559 2793935 27915598-279393 Os06g02051 Os04g05575 531516353168 53151 53168 5315163-5316 27915598279393 279155 279393 27915598-27939 Os06g0205 Os04g0557 53151635316 5315 5316 5315163-531 2791559827939 27915 27939 27915598-2793 Os06g020 Os04g055 5315163531 531 5315163-53 279155982793 2791 2793 27915598-279 Os06g02 Os04g05 531516353 53 5315163-5 27915598279 279 27915598-27 Os06g0 Os04g0 53151635 5 5315163- 2791559827 27 27915598-2 Os06g Os04g 279155982 2 27915598-

Abstract Fluoroquinolones are an important class of antimicrobial agents to manage infectious diseases. However, knowledge about how host bile acids are modified by fluoroquinolones is limited. We investigated and compared the impact of fluoroquinolones on circulating bile acid profiles and gut microbiota from in vivo studies. We administered ciprofloxacin (100 mg/kg/day) or moxifloxacin (40 mg/kg/day) orally to male Wistar rats for seven days. Fifteen bile acids (BAs) from the serum and large intestine were quantified by HPLC-MS/MS. The diversity of gut microbiota after ciprofloxacin and moxifloxacin treatment was analyzed using high-throughput, next-generation sequencing technology. The two fluoroquinolone-treated groups had different BA profiles. Ciprofloxacin significantly reduced the hydrophobicity index of the BA pool, reduced secondary BAs, and increased taurine-conjugated primary BAs in both the serum and large intestine as compared with moxifloxacin. Besides, ciprofloxacin treatment altered intestinal microbiota with a remarkable increase in Firmicutes to Bacteroidetes ratio, while moxifloxacin exerted no effect. What we found suggests that different fluoroquinolones have a distinct effect on the host BAs metabolism and intestinal bacteria, and therefore provide guidance on the selection of fluoroquinolones to treat infectious diseases.

Abstract The aim of this study was to determine the resistance mechanism of Pseudomonas aeruginosa to cefoperazone sodium/sulbactam sodium. We retrospectively analyzed the drug resistance of P.a isolated at the First Affiliated Hospital of Guangxi Medical University. Drug-resistant P.a strains were constructed, then wild-type (WT) and drug-resistant (DR) strains were compared using protein and gene microarrays to determine differences between DR and WT strains. The resistance rates of P. aeruginosa during 2013, 2014 and 2015 were 21.2%, 21.4%, and 24.6% respectively. Among 242 protein peaks of WT and DR bacteriophage proteins, 41 were differentially expressed between the two groups. The expression of 26 and 15 proteins were respectively upregulated and downregulated in the DR compared with the WT group. Gene microarray results revealed 679 mutant loci in the DR group, of which 42 with the top 50 Q values were found in the NCBI database. The rate of P.a resistance to cefoperazone sodium/sulbactam sodium remained high between 2013 and 2015. The numbers of different proteins and genetic variations in the DR strains suggested that the resistance mechanism of P.a to cefoperazone sodium/sulbactam sodium involves multiple genes and proteins that might be key to controlling P.a resistance to cefoperazone sodium/sulbactam sodium.

Abstract Paired basic amino acid-cleaving enzyme 4 (PACE4), a proprotein convertase, is involved in the activation of aggrecanases (ADAMTS-4 and ADAMTS-5) in osteoarthritic and cytokine-stimulated cartilage. Activated aggrecanases cause aggrecan degradation and thus, contribute to osteoarthritis (OA). In this study, we investigated the association between PACE4 gene polymorphisms and OA risk. One single-nucleotide polymorphism (rs4965833) in the PACE4 gene was genotyped in 432 OA patients and 523 healthy controls using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Quantitative reverse transcription PCR (qRT-PCR) was used to determine the relative expression of PACE4 in blood samples from 90 OA patients (30 for each genotype). The relative expression level of PACE4 mRNA was higher in the GG genotype as compared to the AA/AG group. Moreover, the PACE4 rs4965833 polymorphism was associated with increased risk of OA, especially among individuals aged ≥55 years and with a body mass index ≥25. There was no significant association between the PACE4 rs4965833 polymorphism and clinical parameters of OA patients, such as erythrocyte sedimentation rate, C-reactive protein, Visual Analog Scale for pain and Lequesne’s index. In conclusion, the rs4965833 polymorphism in the 3’-UTR of PACE4 is associated with OA susceptibility.

Abstract The goal of this research was to investigate the linkage disequilibrium between rs9263726 and HLA-B*58:01 in different Chinese ethnic groups (Han, Tibet, and Hui) and to study the feasibility of rs9263726 replacing HLA-B*58:01 as an efficient indicator of potential allopurinol hypersensitivity syndrome. In this study, rs9263726 and HLA-B*58:01 were detected in all samples. For samples of individuals whose rs9263726 genotypes were not consistent with HLA-B*58:01, we did high-resolution typing of HLA-B gene to further confirm the correlation of rs9263726 genotype and special HLA-B alleles. We confirmed that the linkage disequilibrium between rs9263726 and HLA-B*58:01 was more significant in the Han ethnic group (r2=0.886, D’=1.0) than in the Tibet and Hui ethnic groups (for Tibetan, r2=0.606, D’=0.866; for Hui, r2=0.622, D’=0.924). For Han Chinese, samples with the GG genotype of rs9263726 did not carry HLA-B*58:01, while AA genotype samples were homozygous carriers of HLA-B*58:01. However, GA genotype samples of rs9263726 required a more sophisticated HLA-B genotyping assay before it was possible to identify whether they were HLA-B*58:01 carriers or not. For Tibetan and Hui, the linkage disequilibrium between rs9263726 and HLA-B*58:01 was not significant. Therefore, rs9263726 cannot replace HLA-B*58:01 in these two groups.

ABSTRACT To seek a simple, rapid and sensitive Coprinus cinereus Peroxidase (CIP) activity assay, a convenient one-factor-at-a-time (OFAT) method and a response surface methodology (RSM) were used. The recombinant CIP expressed in Pichia pastoris was purified with the Ni-NTA spin column. Based on the results of catalytic efficiency (kcat/Km) analysis, 2,2'-azinobis (ethylbenzthiazoline -6-sulfonate) (ABTS) was selected as the optimal enzyme substrate. Results of the OFAT method showed that enzymatic reaction performed in 0.1 mol/L sodium acetate (pH 5.0) buffer in a 200-µl reaction mixture containing 0.5 mmol/L ABTS, 10 mmol/L hydrogen peroxide (H2O2), 49.7 ng CIP at 25°C gave an average CIP activity of 88 U/mL. The ABTS and H2O2 concentrations were then further optimized to improve the sensitivity of the assay. To do that, RSM was conducted through central composite design, and a reduced quadratic model with good fit regression equation was generated. ANOVA analysis of this model indicated that the concentrations of ABTS and H2O2 and their interaction had significant impact on the assay sensitivity. The optimal reaction mixture was determined to include an initial ABTS concentration of 0.82 mmol/L 49.7 ng CIP and 16.36 mmol/L H2O2, and the activity under this condition was determined to be 138.89 U/mL.

Abstract The principal objective of this study was to evaluate the kinetics of dihydroxyacetone production by Gluconobacter frateurii CGMCC 5397 under different oxygen volumetric mass transfer coefficient (kLa) conditions in submerged bioreactors using biodiesel-derived crude glycerol as the carbon source. kLa is a key fermentation parameter for the production of dihydroxyacetone. Cultivations were conducted in baffled- and unbaffled-flask cultures (the kLa values were 24.32 h−1 and 52.05 h−1, respectively) and fed-batch cultures (the kLa values were held at 18.21 h−1, 46.03 h−1, and 82.14 h−1) to achieve high dihydroxyacetone concentration and productivity. The results showed that a high kLa could dramatically increase dihydroxyacetone concentrations and productivities. The baffled-flask culture (with a kLa of 52.05 h−1) favored glycerol utilization and dihydroxyacetone production, and a dihydroxyacetone concentration as high as 131.16 g/L was achieved. When the kLa was set to 82.14 h−1 in the fed-batch culture, the dihydroxyacetone concentration, productivity and yield were 175.44 g/L, 7.96 g/L/h and 0.89 g/g, respectively, all of which were significantly higher than those in previous studies and will benefit dihydroxyacetone industrial production.

BACKGROUND: Ubiquitin Specific Peptidase 39 (USP39) is a 65 kDa SR-related protein involved in RNA splicing. Previous studies showed that USP39 is related with tumorigenesis of human breast cancer cells. RESULTS: In the present study, we investigated the functions of USP39 in human hepatocellular carcinoma (HCC) cell line SMMC-7721. We knocked down the expression of USP39 through lentivirus mediated RNA interference. The results of qRT-PCR and western blotting assay showed that both the mRNA and protein levels were suppressed efficiently after USP39 specific shRNA was delivered into SMMC-7721 cells. Cell growth was significantly inhibited as determined by MTT assay. Crystal violet staining indicated that colony numbers and sizes were both reduced after knock-down of USP39. Furthermore, suppression of USP39 arrested cell cycle progression at G2/M phase in SMMC-7721cells. In addition, Annexin V showed that downregulation of USP39 significantly increased the population of apoptotic cells. CONCLUSIONS: All our results suggest that USP39 is important for HCC cell proliferation and is a potential target for molecular therapy of HCC.

BACKGROUND: Hepcidin, encoding by HAMP gene, is the pivotal regulator of iron metabolism, controlling the systemic absorption and transportation of irons from intracellular stores. Abnormal levels of HAMP expression alter plasma iron parameters and lead to iron metabolism disorders. Therefore,itis animportant goal to understand the mechanisms controlling HAMP gene expression. RESULTS: Overexpression of Sox2 decrease basal expression of HAMP or induced by IL-6 or BMP-2, whereas, knockdown of Sox2 can increase HAMP expression, furthermore, two potential Sox2-binding sites were identified within the human HAMP promoter. Indeed, luciferase experiments demonstrated that deletion of any Sox2-binding site impaired the negative regulation of Sox2 on HAMP promoter transcriptional activity in basal conditions. ChIP experiments showed that Sox2 could directly bind to these sites. Finally, we verified the role of Sox2 to negatively regulate HAMP expression in human primary hepatocytes. CONCLUSION: We found that Sox2 as a novel factor to bind with HAMP promoter to negatively regulate HAMP expression, which may be further implicated as a therapeutic option for the amelioration of HAMP-overexpression-related diseases, including iron deficiency anemia.

OBJECTIVE: The role of Ulinastatin in neuronal injury after cardiopulmonary resuscitation has not been elucidated. We aim to evaluate the effects of Ulinastatin on inflammation, oxidation, and neuronal injury in the cerebral cortex after cardiopulmonary resuscitation. METHODS: Ventricular fibrillation was induced in 76 adult male Wistar rats for 6 min, after which cardiopulmonary resuscitation was initiated. After spontaneous circulation returned, the rats were split into two groups: the Ulinastatin 100,000 unit/kg group or the PBS-treated control group. Blood and cerebral cortex samples were obtained and compared at 2, 4, and 8 h after return of spontaneous circulation. The protein levels of tumor necrosis factor alpha (TNF-α) and interleukin 6 (IL-6) were assayed using an enzyme-linked immunosorbent assay, and mRNA levels were quantified via real-time polymerase chain reaction. Myeloperoxidase and Malondialdehyde were measured by spectrophotometry. The translocation of nuclear factor-κB p65 was assayed by Western blot. The viable and apoptotic neurons were detected by Nissl and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL). RESULTS: Ulinastatin treatment decreased plasma levels of TNF-α and IL-6, expression of mRNA, and Myeloperoxidase and Malondialdehyde in the cerebral cortex. In addition, Ulinastatin attenuated the translocation of nuclear factor-κB p65 at 2, 4, and 8 hours after the return of spontaneous circulation. Ulinastatin increased the number of living neurons and decreased TUNEL-positive neuron numbers in the cortex at 72 h after the return of spontaneous circulation. CONCLUSIONS: Ulinastatin preserved neuronal survival and inhibited neuron apoptosis after the return of spontaneous circulation in Wistar rats via attenuation of the oxidative stress response and translocation of nuclear factor-κB p65 in the cortex. In addition, Ulinastatin decreased the production of TNF-α, IL-6, Myeloperoxidase, and Malondialdehyde.

Background: Finding molecular markers linked to quantitative trait loci is the first step in marker-assisted selection (MAS). Microsatellites are excellent molecular markers because of their large numbers, even distribution in the genome, and high polymorphism. In this study, the polymerisation effect of four microsatellites (OarAE101, BM1329, BM143, and LSCV043) on litter size was analysed using microsatellite markers and pedigrees. Results: The results indicate that the polymerisation effect of four microsatellite loci significantly affected the litter size. E5E10F2F6G1G5H6H11 and E3E8F5F7G1G5H3H9 had the highest and lowest litter sizes in the F2 generation, respectively. The polymerisation effect value (v) of the E5E10 genotype was 3.18% higher than that of the E2E7 genotype. The v of genotype F2F6 was 14.47% higher than that of the F5F7 genotype. The v of genotype G1G5 was 58.99% higher than that of the G2G7 genotype. The v of the H6H11 genotype was 5.60% to 49.74% higher than those of the H4H10 and H1H7 genotypes. The v of the H3H9 genotype was 17.22% higher than that of the H1H7 genotype. Conclusions: The results of the present study are vital to improving the reproductive performance in goat breeds MAS.

OBJETIVOS: A reconstrução pélvica após excisão de tumor é um desafio. MÉTODOS: Realizou-se um estudo retrospectivo para comparar os desfechos entre pacientes submetidos a cirurgia de reconstrução da pelve com aloenxerto ósseo após excisão em bloco de tumores pélvicos e pacientes submetidos apenas à excisão. RESULTADOS: Os pacientes sem reconstrução tiveram escores funcionais significantemente menores 3 meses (10 vs. 15, P = 0,001) e 6 meses após a cirurgia (18,5 vs. 22, P = 0,0024), menor tempo de hospitalização (16 dias vs. 40 dias, P < 0,001) e menor custo hospitalar (97.500 vs. 193.000 yuans, P < 0,001) do que os que foram submetidos a reconstrução pélvica. Os escores funcionais foram similares 12 meses depois da cirurgia (21,5 vs. 23, P = 0,365) sem diferença na taxa de complicações entre os dois grupos (P > 0,05). CONCLUSÕES: A reconstrução pélvica com aloenxerto ósseo depois de cirurgia de tumores pélvicos é associada a desfechos cirúrgicos e funcionais satisfatórios. Outros estudos clínicos são necessários para explorar como selecionar o melhor método de reconstrução. Nível de evidência iv, séries de casos.

OBJECTIVES: Pelvic reconstruction after tumor resection is challenging. METHODS: A retrospective study had been performed to compare the outcomes between patients undergoing reconstructive surgery of the pelvis with allogeneic bone graft after en bloc resection of pelvic tumors and patients undergoing en bloc resection only. RESULTS: Patients without reconstruction had significantly lower functional scores at 3 months (10 vs. 15, P = 0.001) and 6 months after surgery (18.5 vs. 22, P = 0.0024), a shorter duration of hospitalization (16 days vs. 40 days, P < 0.001), and lower hospital costs (97,500 vs. 193,000 RMB, P < 0.001) than those undergoing pelvic reconstruction. Functional scores were similar after 12 months of surgery (21.5 vs. 23, P = 0.365) with no difference in the rate of complications between the two groups (P > 0.05). CONCLUSIONS: Pelvic reconstruction with allogeneic bone graft after surgical management of pelvic tumors is associated with satisfactory surgical and functional outcomes. Further clinical studies are required to explore how to select the best reconstruction method. Level of evidence iv, case series.