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Antibacterials represent a pharmaceutical class that is extensively used and consumed worldwide. The presence of a large number of antibacterial agents in water could result in antibiotic resistance. Thus, the development of a fast, accurate, and high-throughput method to analyze these emerging contaminants in water is necessary. Herein, a method was developed to achieve the simultaneous determination of 43 antibacterials from nine pharmaceutical categories (i.e., sulfonamides, quinolones, fluoroquinolones, tetracyclines, lincosamides, macrolides, nitroimidazoles, diterpenes, and dihydrofolate reductase inhibitors) in water using automatic sample loading-solid phase extraction (SPE)-ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Because the properties of these 43 antibacterials are quite different, the main objective of this work is to develop an extraction procedure that would enable the simultaneous analysis of a wide range of multiclass antibacterials. Given this context, the work presented in this paper optimized the SPE cartridge type, pH, and sample loading amount. Multiresidue extraction was performed as follows. The water samples were filtered through 0.45 μm filter membranes, added with Na 2 EDTA and NaH 2 PO 4 , and pH-adjusted to 2.34 using H 3 PO 4 . The solutions were then mixed with the internal standards. An automatic sample loading device fabricated by the authors was used for sample loading, and Oasis HLB cartridges were used for enrichment and purification. The optimized UPLC conditions were as follows: chromatographic column, Waters Acquity UPLC BEH C 18 column (50 mm×2.1 mm, 1.7 μm); mobile phases, methanol-acetonitrile (2∶8, v/v) solution containing 0.1% formic acid and 0.1% formic acid aqueous solution; flow rate, 0.3 mL/min; injection volume, 10 μL. The compounds were step scanned using an electrospray ionization source in the positive and multiple-reaction monitoring (MRM) modes, and analyzed by internal and external standard methods. The results showed that the 43 compounds achieved high linearity in their respective linear ranges, with correlation coefficients ( r 2 ) greater than 0.996. The limits of detection (LODs) of the 43 antibacterial agents ranged from 0.004 ng/L to 1.000 ng/L, and their limits of quantification (LOQs) ranged from 0.012 ng/L to 3.000 ng/L. The average recoveries ranged from 53.7% to 130.4%, and the relative standard deviations (RSDs) were between 0.9% and 13.2%. The method was successfully applied to the determination of six tap water samples from different districts and six water samples obtained from the Jiangyin section of the Yangtze River and Xicheng Canal. No antibacterial compound was detected in any of the tap water samples, but a total of 20 antibacterial compounds were detected in the river and canal water samples. Among these compounds, sulfamethoxazole showed the highest mass concentrations, ranging from 8.92 to 11.03 ng/L. The types and contents of antibacterials detected in the Xicheng Canal were greater than those found in the Yangtze River, and two kinds of diterpenes, namely tiamulin and valnemulin, were found easily and commonly in water sample. The findings indicate that antibacterial agents are widespread in environmental waters. The developed method is accurate, sensitive, rapid, and suitable for the detection of the 43 antibacterial compounds in water samples.

Objective: To explore the key deubiquitinating enzymes that maintain the stemness of liver cancer stem cells and provide new ideas for targeted liver cancer therapy. Methods: The high-throughput CRISPR screening technology was used to screen the deubiquitinating enzymes that maintain the stemness of liver cancer stem cells. RT-qPCR and Western blot were used to analyze gene expression levels. Stemness of liver cancer cells was detected by spheroid-formation and soft agar colony formation assays. Tumor growth in nude mice was detected by subcutaneous tumor-bearing experiments. Bioinformatics and clinical samples were examined for the clinical significance of target genes. Results: MINDY1 was highly expressed in liver cancer stem cells. The expression of stem markers, the self-renewal ability of cells, and the growth of transplanted tumors were significantly reduced and inhibited after knocking out MINDY1, and its mechanism of action may be related to the regulation of the Wnt signaling pathway. The expression level of MINDY1 was higher in liver cancer tissues than that in adjacent tumors, which was closely related to tumor progression, and its high expression was an independent risk factor for a poor prognosis of liver cancer. Conclusion: The deubiquitinating enzyme MINDY1 promotes stemness in liver cancer cells and is one of the independent predictors of poor prognosis in liver cancer.

Introduction: Portal vein system thrombosis (PVST) is a common, potentially life-threatening complication after splenectomy. The optimal recognized anticoagulation drugs for preventing PVST in cirrhotic patients after splenectomy remain unclear. The aim of this study is to evaluate the safety and efficacy of warfarin in preventing PVST after laparoscopic splenectomy and azygoportal disconnection (LSD)., Methods: In this randomized controlled single-center study, 80 cirrhotic patients who underwent LSD were randomly assigned to 2 years of treatment with either warfarin (n = 40) or aspirin (n = 40). The primary outcome was prevention of PVST. Sonographers and radiologists who assessed outcomes were blinded to group assignments. Intention-to-treat analysis was performed., Results: During the first year, excluding two patients withdrawing from the study, 15 of the 39 warfarin-treated patients (38.5%) and five of the 39 aspirin-treated patients (12.8%) did not develop PVST (P = 0.010). The incidence of PVST in the first 2 postoperative years was significantly lower in the warfarin group than in the aspirin group (F = 7.360, P = 0.008). The warfarin group in paired within-group comparisons had significantly greater improvements in total bilirubin and albumin levels at baseline versus at 6 months postoperatively and in creatinine levels at baseline versus at 12, 18, and 24 months postoperatively respectively (all P < 0.05). In contrast, those paired comparisons of the aspirin group showed no significant differences (all P > 0.05)., Conclusions: Warfarin therapy was safe and effective and significantly reduced the risk of PVST after LSD, compared with aspirin treatment. Warfarin treatment was associated with better liver function protection and renal function improvement than aspirin treatment., (Copyright © 2019 IJS Publishing Group Ltd. Published by Elsevier Ltd. All rights reserved.)

The recent success of derivation of mammalian haploid embryonic stem cells (haESCs) has provided a powerful tool for large-scale functional analysis of the mammalian genome. However, haESCs rapidly become diploidized after differentiation, posing challenges for genetic analysis. Here, we show that the spontaneous diploidization of haESCs happens in metaphase due to mitotic slippage. Diploidization can be suppressed by small-molecule-mediated inhibition of CDK1 and ROCK. Through ROCK inhibition, we can generate haploid somatic cells of all three germ layers from haESCs, including terminally differentiated neurons. Using piggyBac transposon-based insertional mutagenesis, we generated a haploid neural cell library harboring genome-wide mutations for genetic screening. As a proof of concept, we screened for Mn 2+ -mediated toxicity and identified the Park2 gene. Our findings expand the applications of mouse haploid cell technology to somatic cell types and may also shed light on the mechanisms of ploidy maintenance., (Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.)

The epithelial-to-mesenchymal transition (EMT) is a critical step in tumor metastasis. NEDD9 has been shown to be an oncogene in colorectal cancer. However, little is known about the relationship between NEDD9 and EMT in colorectal cancer metastasis. A total of 63 pairs of freshly frozen colorectal cancer tissues and adjacent noncancerous tissues were evaluated for NEDD9 gene expression using quantitative real-time PCR. The expression of NEDD9 and three epithelial-mesenchymal transition (EMT)-related proteins (E-cadherin, β-catenin and vimentin) was examined in 122 colorectal cancers by immunohistochemistry. The expression of NEDD9 was markedly increased in colorectal cancer tissues compared with adjacent noncancerous tissues. The expression level of NEDD9 was positively correlated and TNM stage but not with other clinicopathological features of colorectal tumors. Furthermore, the expression of NEDD9 was strongly associated with the loss of epithelial marker E-cadherin and acquired expression of the mesenchymal markers nuclear β-catenin and vimentin. These findings suggested that NEDD9 might promote EMT and the progression of colorectal cancer, and thus may be a potential therapeutic target of colorectal cancers., Competing Interests: None., (IJCEP Copyright © 2017.)

Introduction: Laparoscopic splenectomy and azygoportal disconnection (LSD) has been reported to be safe, feasible, and minimally invasive for cirrhotic patients with portal hypertension. There is still controversy as to whether it is necessary to perform synchronous splenectomy for patients with moderate hypersplenism who undergo azygoportal disconnection for esophagogastric variceal hemorrhage (EGVB)., Methods: We retrospectively evaluated the outcomes in 51 cirrhotic patients with EGVB and moderate hypersplenism (PLT ≥50 × 10 9 /L) who underwent LSD (n = 28) or laparoscopic azygoportal disconnection (LD) (n = 23) between January 2014 and October 2015. Their demographic, intraoperative, and postoperative variables were compared., Results: LSD and LD were successful in all the patients. When compared with LSD, LD had a significantly shorter operation time, less intraoperative blood loss, shorter postoperative hospital stay, fewer days of postoperative body temperature >38.0 °C, lower rate of fever postoperatively, and lower C-reactive protein concentration and procalcitonin concentration on postoperative day (POD) 7 (all P < 0.05). The incidences of portal vein system thrombosis in the LD group on PODs 7, 30, and 90 were significantly lower than those in the LSD group at all the time points (all P < 0.05). According to the postoperative serum proportions of CD4 + and CD8 + and the CD4+/CD8+ ratio (all P < 0.05), the LSD group had significantly lower immune function than the LD group on POD 90., Conclusions: LD is safe and effective for EGVB with moderate hypersplenism secondary to portal hypertension in selected patients., (Copyright © 2016 IJS Publishing Group Ltd. Published by Elsevier Ltd. All rights reserved.)

Background: Portal vein system thrombosis (PVST) is a frequent and potentially life-threatening complication after laparoscopic splenectomy and azygoportal disconnection (LSD) in patients with cirrhotic portal hypertension. The objective of this study was to investigate the safety and effectiveness of warfarin with a target international normalized ratio (INR) of 2.0-2.5 for the prevention of PVST after LSD. Hitherto, this is the first study to assess the use of warfarin in this field., Materials and Methods: We retrospectively analyzed a database of 73 consecutive patients who underwent LSD from January 2013 to September 2014. Patients were categorized into the warfarin group (34 patients) and the aspirin group (39 patients). The INR and incidence of PSVT were monitored for 90 days., Results: Compared with the aspirin group, the warfarin group had a lower incidence of PVST on postoperative day (POD) 30 [17/34 (50.0%) versus 29/39 (74.4%); P = .032] and POD 90 [8/34 (23.5%) versus 30/39 (76.9%); P < .0001] and main portal vein thrombosis (MPVT) on POD 90 [3 (8.8%) versus 13 (33.3%); P = .012]. From POD 30 to 90, the warfarin group achieved more complete recanalization of PVST [9/17 (52.9%) versus 3/29 (10.3%), P = .005] and MPVT [9/12 (75.0%) versus 3/12 (25.0%), P = .039]. Multiple logistic regression analysis revealed that warfarin was an independent protective factor for PVST at POD 90 (relative risk, 0.027; 95% confidence interval, 0.004-0.168; P < .001). No patients developed bleeding complications., Conclusions: Anticoagulation therapy with warfarin is safe and effective for the prevention of PVST in cirrhotic patients with portal hypertension after LSD.

Introduction: Portal vein system thrombosis (PVST) is an alarming and potentially life-threatening complication of laparoscopic splenectomy and azygoportal disconnection (LSD). The objective of this study was to investigate negative and positive predictors of PVST after LSD in patients receiving anticoagulant regimens with aspirin or warfarin., Methods: Seventy-five consecutive patients who underwent LSD from 2013 to 2014 were retrospectively reviewed. Patients received anticoagulant regimen with warfarin (n = 35) or aspirin (n = 40) according to individual preference. International normalized ratio (INR) and the incidence of PSVT were compared in patients received anticoagulant regimen with warfarin or aspirin on postoperative days (POD) 7, 30, and 90, and factors associated with PVST at these time points were determined by univariate and logistic multivariable regression analyses., Results: Portal vein diameter was an independent negative predictor of PVST on PODs 7, 30, and 90. Anticoagulation with warfarin was an independent positive predictor of PVST on PODs 30 and 90, and INR was an independent positive predictor of PVST on POD 90. Dynamic changes in the incidence of PVST on the day of admission and on PODs 7, 30, and 90 differed significantly between the warfarin and aspirin groups (P = 0.002). No patient experienced perioperative bleeding., Conclusions: Portal vein diameter was an independent negative predictor, while anticoagulation therapy with warfarin and INR were independent positive predictors, of PVST after LSD. Early anticoagulation with warfarin is safe and effective for the prevention of PVST after LSD., (Copyright © 2016 IJS Publishing Group Ltd. Published by Elsevier Ltd. All rights reserved.)

The epithelial‑to‑mesenchymal transition (EMT) has been noted as a critical event in the early step of cancer metastasis. Recent studies showed that nuclear transcription factor caudal type homeobox transcription factor 2 (CDX2) is a prognostic factor, which acts as a marker of good outcome in gastric cancer (GC) patients. However, the association between CDX2 expression and EMT has remained to be fully elucidated. The present study reported that forced overexpression of CDX2 in MKN45/CDX2 cells inhibited GC‑cell growth and proliferation, and attenuated migration and invasion in vitro. Furthermore, MKN45/CDX2 cells exhibited a significant upregulation of E‑cadherin protein and a significant downregulation of vimentin protein expression. These results were further supported by in vivo tumorigenicity assays, which showed that CDX2 suppressed gastric tumor xenograft growth and inhibited EMT in nude mice. These results indicated that CDX2 is capable of inhibiting GC‑cell growth and invasion. CDX2 may participate in the process of EMT of GC cells by regulating the expression of the epithelial and mesenchymal proteins E‑cadherin and vimentin.

Background: Cases of primary gastric adenocarcinoma with metastasis to the breast are extremely rare. Till now, only 38 cases have been reported in PubMed since 1908., Case Presentation: We herein reported a race case of gastric adenocarcinoma with metastasis to the right breast. Breast biopsy showed invasive signet-ring cell breast carcinoma in the right breast. She was given a TEC regimen (docetaxel 75 mg/m(2), epirubicin 75 mg/m(2), and cyclophosphamide 600 mg/m(2) every 3 weeks) for one cycle but showed no objective response. Upper gastrointestinal endoscopy demonstrated an ulcerative mass in the gastric body. Biopsy demonstrated low-grade gastric adenocarcinoma with signet-ring features. In immunohistochemistry, mammary malignant cells were positive for cytokeratin 7, cytokeratin 20, villin, and ErbB2/HER2, but negative for gross cystic disease fluid protein-15, estrogen receptor, and progesterone receptor. The diagnosis of metastatic poorly differentiated signet-ring cell adenocarcinoma of the right breast identical to gastric primary was confirmed finally., Conclusions: Gastric cancer with metastasis to the breast can be diagnosed by clinical history, histological findings, and immunohistochemical markers.

Pluripotent stem cells can be evaluated by pluripotent markers expression, embryoid body aggregation, teratoma formation, chimera contribution and even more, tetraploid complementation. Whether iPS cells in general are functionally equivalent to normal ESCs is difficult to establish. Here, we present the detailed procedure for chimera formation and tetraploid complementation, the most stringent criterion, to assessing pluripotency.

The rocket kerosene quality properties such as density, distillation range, viscosity and iodine value were successfully measured based on their near-infrared spectrum (NIRS) and chemometrics. In the present paper, more than 70 rocket kerosene samples were determined by near infrared spectrum, the models were built using the partial least squares method within the appropriate wavelength range. The correlation coefficients (R2) of every rocket kerosene's quality properties ranged from 0.862 to 0.999. Ten unknown samples were determined with the model, and the result showed that the prediction accuracy of near infrared spectrum method accords with standard analysis requirements. The new method is well suitable for replacing the traditional standard method to rapidly determine the properties of the rocket kerosene.

The Wnt signaling pathway is thought to be functionally conserved in vertebrates and invertebrates and plays an important role during the embryonic and postembryonic development. Recent studies indicated that this pathway may be also involved in the controlled proliferation and migration of some kinds of fibroblasts during the wound healing process. To verify this assumption in vitro, we chose Rat-1, a kind of rat fibroblasts to investigate the regulation of Wnt signaling pathway to the growth and changes of several phenotypes of this kind of cells. Full length Wnt-3a cDNA was inserted in pcDNA 3.1 vector to construct the Wnt-3a mammalian expression vector, which was stably transfected into Rat-1 cells, and then to establish a cell model in which Wnt signaling pathway was constantly activated. When Wnt signaling pathway was activated constantly, Rat-1 cells exhibited morphological changes: grew more densely as a monolayer, adopted an elongated and refractile appearance, forming cord-like bundles lined up in a uniform direction. The results of MTT assay and FCM analysis indicated that more Rat-1/Wnt-3a cells entered into G(2) phase and the proliferation rate of the Rat-1/ Wnt-3a cells increased significantly compared to the non-transfected cells. Though the migration of Rat-1/Wnt-3a cells increased slightly by the method of Transwell migration assay, there was no statistic significance compared to the non-transfected cells. The result of in vitro scrape wound healing assay showed that for Rat-1/Wnt-3a cells the time course of wound healing decreased significantly. It is therefore concluded that the activation of Wnt signaling pathway can regulate some of the phenotypes of Rat-1 cells, facilitate cell proliferation and promote the scrape wound healing in vitro.

The fluorescence and synchronous fluorescence spectra were employed to study the binding between sodium azide (NaN3) and horseradish peroxidase (HRP), which was affected by the molecular conformation and the microenvironment of the fluorescence residues. The mechanism of the fluorescence quenching of HRP by NaN3 was discussed and the binding constant and the number of binding sites of non-covalent bond between them were calculated.

Comparing the fluorescence spectra of tobacco peroxidase I (TOP I) solution and the solutions titrated by iodine anions, the number of binding locus and the binding constant of iodine anion to TOP I were calculated by using the modified Stern-Volmer equation. The mechanism of the quenching of fluorescence spectra and the distribution of tryptophane residues in the TOP I molecule were discussed.

The ultraviolet/visible spectra of TOP I were studied. It was proved that TOP I is an acid enzyme containing it hemochrome as an agon. When the pH reduced, the Soret absorption in UV-Vis region exhibited a blue shift, when pH increased exhibited a red shift. The result of the influence of carbamide, the denaturant, on the spectra showed that TOP I may have a unfolded structural change in the solution, which made the peptide chain completely extend. After adding Fe(III), Fe(II), Cu(II), Zn(II), Co(II), Ni(II) and Sn(II) to the apo-TOP I, the UV-Vis spectra changed except for Fe(III), which almost did not change at all. The strongest characteristic absorption peak of the Soret showed a blue shift to different extent and it became weak gradually. The situation of alpha-strip and beta-strip remained unchanged while the relative strength decreased.

Tobacco peroxidase (TOPI) has been purified by ammonium sulfate fractionation and column chromatography, involving ion exchange on DE-52 cellulose, gelfiltration on Sephadex G-75 and ion exchange on DEAE-sephadex A-50. The specific activity of peroxidase purified was 4,826 U/mg. It has been demonstrated by SDS-PAGE as a single band. Its molecular weight (matrix assisted laser desorption/ionization time of flight mass spectra) and isoelectric point (pI) have been determined to be 21,888.5 and 3.5 respectively. The native enzyme is one of a haemachrome-containing acidic protein and has its soret maximum at 402 nm and alpha and beta bands at 636 nm and 498 nm, respectively. Its characteristic absorption spectra and fluorescence spectra changed in different pH and denaturant. It has its own characteristic absorption spectra and fluorescence spectra.

A new ion chromatographic method was developed for the quantitative determination of ammonia and potassium in tobacco. The tobacco samples were treated with dilute nitric acid and K+, NH4+ were extracted. The method is fast and convenient. The relationships between peak heights and initial concentrations of K+ and NH4+ were linear. The detection limits were 0.05 mg/L for K+ and 0.03 mg/L for NH4+. Recoveries of K+ and NH4+ were 96.7%-106.7% and 96.5%-109.1% respectively. The whole procedure of determination can be completed within 5 min.

The present study aimed to explore modulation of ATP-activated currents (I(ATP)) by met-Enk in rat DRG neurons. I(ATP) was recorded using the whole-cell patch clamp technique. The majority of the neurons examined responded to ATP (90.0 percent;, 45/50) with inward currents. In the 45 ATP sensitive neurons three kinds of responses to application of met-Enk were distinguished: (l) inward currents (29/45), (2) outward currents (9/45), and (3) no effect (7/45). Pretreatment with met-Enk (10(-9)~10(-5) mol/L) suppressed I(ATP) (10(-4) mol/L) in 29 neurons responding to met-Enk with inward currents. The inhibition by met-Enk of I(ATP) could be blocked by naloxone (10(-7) mol/L) in a concentration-dependent manner. Met-Enk of 10(-9), 10(-8), 10(-7), 10(-6) and 10(-5) mol/L suppressed I(ATP) by l3.2+/-5.4 percent; (n=5); 39.2+/-8.6 percent; (n=8), 54.l+/-8.6 percent; (n=8),43.3+/-7.9 percent; (n=7) and 43.l+/-7.9 percent; (n=7) (mean+/-MSE), respectively. A comparison of concentration - response relations of ATP with and without preapplication of met-Enk indicated that after pretreatment with met-Enk (10(-7) mol/L) the curve shifted downward markedly with a decrease of 25 percent; of the maximum value of I(ATP) and unchanged K(d) value. The suppression of I(ATP) by met-Enk was reversed as evidenced by intracellular dialysis of H-9 by using the repatch technique. Taken together, it is suggested that the inhibition by met-Enk of I(ATP) is caused by activation of opiate receptor, which eventually results in phosphorylation of ATP receptor, mediated by modulation of G protein coupling and intracellular signal transduction.

To elucidate the functions of grp75, CHL cells over-expressing grp75 are cultured in glucose-free medium in order to simulate energy metabolic stress. Their susceptibilities to injuries caused by glucose deprivation are assessed by trypan blue exclusion, LDH leakage measurement and cytometry analysis. Data shows a stronger resistance to glucose deprivation among cells over-expressing grp75 than among cells constitutively expressing grp75. The outcome suggests that grp75 can protect cells from injuries caused by glucose deprivation.

Experiments were performed on 55 urethane anesthetized and flaxedil immobilized rabbits under artificial respiration. Injection of L-Glutamate into the locus coeruleus area led to a marked augmentation of respiration. This effect of L-Glutamate could be significantly attenuated or reversed by a previous injection of prazosin, yohimbine and propranolol into the NTS area. These results indicate that the neurons in the locus coeruleus are involved in the regulation of the respiration as a result of mediation by alpha and beta receptors in the NTS area.

The experiments were performed on 55 rabbits vagotomied, anesthetized with urethane and immobilized with flaxedil. Injection of L-glutamate (L-glu) into the locus coeruleus complex (Lc-Sc) area led to a marked increase in respiratory frequency (RF) and phrenic nerve discharge rate (phr.d) (16 +/- 3.5% and 6.5 +/- 2.5%; P < 0.01 and P < 0.05, respectively), a decrease in inspiratory and expiratory duration, and no obvious change in blood pressure (Bp) was observed. While prazosin (0.5 micrograms/microliters), yohimbin (4 micrograms/microliters) and propranolol (2 micrograms/microliters) were preinjected into the nuclei tractus solitraii (NTS) respectively, the above-mentioned effects of L-glu on respiration was blocked. These results indicate that the excitation of neurons in the Lc-Sc can induce an increase in RF and phr. d and the excitative effect of L-glu on RF was mediated by alpha 1, alpha 2 and beta-receptors in the NTS, while the effect on phr.d was mediated differently.