CD14, a novel surface marker of esophageal cancer stem cells.

Studies on targeting cancer stem cells (CSCs) have not yielded satisfactory results regarding solid tumor treatments; one of the reasons for this is the difficulty associated with the identification of a relatively specific antigen in solid tumors. CD14, which is mainly expressed in certain immune cells, is associated with tumor recurrence, growth, metastasis and resistance to treatment, which is in conformity with the characteristics of CSCs. It was thus hypothesized that esophageal CSCs (ECSCs) express CD14. In the present study, paraffin­embedded sections of human esophageal carcinoma were used to determine the co­expression of CD14 and the ECSC marker aldehyde dehydrogenase­1 (ALDH1) using immunofluorescence. CD14+ cells were then isolated using immunomagnetic separation for stemness detection, including proliferation, migration, invasion and tumorigenicity. Cell Counting Kit­8 (CCK­8), EdU and colony­formation assays were utilized to investigate the proliferative ability, the metastatic capacity was examined using Transwell and wound­healing assays and a xenograft assay was performed to investigate the tumorigenic ability. It was indicated that the ALDH1­labeled ECSCs expressed CD14 and primary CD14+ cells possessed the characteristics of CSCs. On the whole, the results of the present study suggest the potential utility of CD14 as a novel surface marker for ECSCs.

Nesfatin-1 protects against diabetic cardiomyopathy in the streptozotocin-induced diabetic mouse model via the p38-MAPK pathway.

Nesfatin-1 is a novel anorexigenic peptide that possesses antihyperglycemic and cardiovascular effects. We hypothesized that nesfatin-1 has a beneficial protective effect against diabetic cardiomyopathy (DC). We investigated the therapeutic effect of nesfatin-1 on diabetes-associated cardiac dysfunction in the high-fat diet (HFD)/streptozotocin (STZ)-induced diabetic mouse model. We found that the cardiac nesfatin-1 level was lower in diabetic mice than in normal mice. Nesfatin-1 treatment (180 mg/kg/day for two weeks) improved insulin sensitivity and mitigated diabetic dyslipidemia. Nesfatin-1 ameliorated the diabetes-related myocardial hypertrophy and heart dysfunction, as revealed by the reduced hypertrophy index, heart rate, mean arterial pressure (MAP), creatine kinase (CK)-MB, and aspartate aminotransferase (AST) levels. Nesfatin-1 exerted antioxidant and anti-inflammatory activity in diabetic mice, as shown by decreased reactive oxygen species (ROS), oxidative lipid product malondialdehyde (MDA) levels, increased superoxide dismutase (SOD) and glutathione (GSH), decreased cardiac and plasma interleukin-1 ß (IL-1ß) and tumor necrosis factor-α (TNF-α) levels. Mechanistically, we found that nesfatin-1 inhibited the cardiac p38-MAPK pathway activation and subsequent glucagon-like peptide-1 (GLP-1) level. Collectively, our data shows nesfatin-1 exerted protective effects against diabetic cardiomyopathy. Our study suggests that nesfatin-1 therapy has therapeutic implications against diabetic cardiomyopathy.

Texture Features of Computed Tomography Image under the Artificial Intelligence Algorithm and Its Predictive Value for Colorectal Liver Metastasis.

The aim of this research was to investigate the predictive role of texture features in computed tomography (CT) images based on artificial intelligence (AI) algorithms for colorectal liver metastases (CRLM). A total of 150 patients with colorectal cancer who were admitted to the hospital were selected as the research objects and randomly divided into three groups with 50 cases in each group. The patients who were found to suffer from the CRLM in the initial examination were included in group A. Patients who were found with CRLM in the follow-up were assigned to group B (B1: metastasis within 0.5 years, 16 cases; B2: metastasis within 0.5-1.0 years, 17 cases; and B3: metastasis within 1.0-2.0 years, 17 cases). Patients without liver metastases during the initial examination and subsequent follow-up were designated as group C. Image textures were analyzed for patients in each group. The prediction accuracy, sensitivity, and specificity of CRLM in patients with six classifiers were calculated, based on which the receiver operator characteristic (ROC) curves were drawn. The results showed that the logistic regression (LR) classifier had the highest prediction accuracy, sensitivity, and specificity, showing the best prediction effect, followed by the linear discriminant (LD) classifier. The prediction accuracy, sensitivity, and specificity of the LR classifier were higher in group B1 and group B3, and the prediction effect was better than that in group B2. The texture features of CT images based on the AI algorithms showed a good prediction effect on CRLM and had a guiding significance for the early diagnosis and treatment of CRLM. In addition, the LR classifier showed the best prediction effect and high clinical value and can be popularized and applied.

Fucoxanthin, a Marine Carotenoid, Suppresses Mycoplasma pneumoniae-Triggered Inflammatory Cytokine Production and Promotes Bacterial Clearance in a Murine Model.

Mycoplasma pneumoniae (MP), an atypical bacterium, is a common pathogenetic organism of respiratory infection in children. In the present study, we analyzed the beneficial role of fucoxanthin (Fx), a marine carotenoid, in a murine model of MP. C57BL/6 mice were inoculated once intranasally with 107 CFU of M. pneumoniae, and we found that Fx treatment markedly decreased BAL (quantitative bronchoalveolar lavage) M. pneumoniae concentrations and alleviated airway obstruction in the infected mice. Moreover, the concentrations of proinflammatory cytokines, including IL-6, TNF-α and IL-1ß, were significantly decreased by Fx treatment in the BAL samples of infected mice. In vitro study further indicated that Fx treatment markedly suppressed the production of proinflammatory cytokines in mouse peritoneal macrophages after M. pneumoniae infection. In conclusion, this may be the first study to report the protective role of Fx against M. pneumoniae infection, providing a potential therapeutic agent for MP.

Effect of Licochalcone-A Combined with Rab23 Gene on Proliferation of Glioma U251 Cells.

This research aimed to explore the effect of Licochalcone-A (LCA) combined with Rab23 gene on the proliferation, migration, and invasion of glioma U251 cells through the Wnt/ß-catenin signaling pathway. The glioma U251 cell line was taken as the research object, and the Rab23 overexpression plasmid was constructed. According to the treatment method, U251 cells were rolled into blank control group (BC), Rab23 overexpression plasmid transfection group (Rab23), 25 µmol·L-1 LCA treatment group (LCA), and Rab23 overexpression plasmid transfection combined with 25 µmol·L-1 LCA treatment group (Rab23 + LCA). Subsequently, the ability of cell proliferation, migration, and invasion of each group was detected by methyl thiazolyl tetrazolium (MTT) assay, scratch healing test, and Transwell cell invasion test, respectively. Western blot was implemented to detect the expression differences of cell proliferation antigen Ki-67, apoptosis-related proteins Bcl-2 and Bax, and Wnt/ß-catenin pathway-related proteins ß-catenin, glycogen synthase kinase-3 (GSK3ß), Axin2, and c-myc. The results showed the successful construction of Rab23 overexpression and stable transfection U251 cell line. After grouping and treatments, the cell proliferation, migration, and invasion ability of the Rab23 group, LCA group, and Rab23 + LCA group was substantially reduced relative to BC group (P < 0.05). In addition, the cell proliferation, migration, and invasion ability of Rab23 + LCA group decreased relatively more significantly. The expression levels of Ki-67, Bcl-2, ß-catenin, and c-myc in the Rab23, LCA, and Rab23 + LCA groups were greatly lower versus those of BC group. Moreover, the protein expression levels of Bax, GSK3ß, and Axin2 were considerably increased (P < 0.05), while the expression of protein in Rab23 + LCA group increased notably. These findings indicate that LCA combined with Rab23 gene can inhibit the proliferation, migration, and invasion of glioma U251 cells through the Wnt/ß-catenin signaling and can promote cell apoptosis.

Naringin Alleviates H2O2-Inhibited Osteogenic Differentiation of Human Adipose-Derived Stromal Cells via Wnt/ß-Catenin Signaling.

Osteoporosis is an age-related systemic bone disease that places a heavy burden on patients and society. In this study, we aimed to investigate the effects of naringin (NAR) on the osteogenic differentiation of human adipose-derived stromal cells (ADSCs). The results demonstrated that NAR pretreatment effectively abated H2O2-induced cell death and ROS accumulation in ADSCs undergoing osteogenic differentiation (ADSCs-OD). In addition, we also observed that the impaired extracellular matrix mineralization and ALP activity in H2O2-stimulated ADSCs-OD were notably rescued by NAR pretreatment. Moreover, the effects of H2O2 exposure on Wnt/ß-catenin signaling in ADSCs-OD were largely reversed by NAR pretreatment. Collectively, our findings indicated that NAR could protect ADSCs-OD against H2O2-inhibited osteogenic differentiation.

[Location and role of protein kinase Cα in parthenogenetic and tetraploid preimplantation embryonic development in mouse].

Protein kinase C (PKC) is a critical molecule in cellular signal transduction in mammals. It is involved in many biological processes in embryonic development, including nuclear remodeling, cell cycle adjustment and cellular polarity regulation. The present study aimed to observe the location of PKCα, an important isozyme of PKC, in fertilized, parthenogenetic and tetraploid preimplantation embryos, and compare the expression of PKCα during embryonic compaction in Kunming mice. The location of PKCα was detected by immunochemistry and laser confocal microscopy. Western blot was performed to quantify PKCα expression during embryonic compaction in the three kinds of embryos. In the experiment, fertilized embryos were flushed from oviduct or uterus at 45, 52, 69, 76 and 93 h after injection of human chorionic gonadotrophin (hCG); parthenogenetic embryos were collected by SrCl2 activation of oocytes for 6 h; and tetraploid embryos were produced by electrofusion of 2-cell embryos. Embryos were fixed at different developmental stages for immunofluorescent staining. 8-cell/4-cell embryos and morula were lysed for Western blot. The results showed that PKCα had similar location pattern in different embryos. It was distributed mainly in the nuclear aggregating around chromatin at different developmental stages. However, PKCα expressed strongly in the interphase than in mitotic blastomere. Before embryonic compaction, PKCα was localized at the blastomere boundary. At late blastocyst stage of fertilized embryos, PKCα was localized only in the polar trophoblast, but not in other trophoblast. At late stage of pathenogenetic blastocyst, there was no clear PKCα signal in the polar trophoblast. Tetraploid embryos had larger blastomere than other embryos and compacted after 4-cell stage, but not after 8-cell stage. Meanwhile, there was PKCα signal at the blastomere boundary at 4-cell stage. Our results showed that the expression of PKCα lasted through all the preimplantation stage. Although there were different expression levels among different stages, the expression increased around embryonic compaction. Quantification of expression of PKCα by Western blot demonstrated that the expression increased after compaction, indicating that the compaction was possibly dependent on the relocation of PKCα. Moreover, it was shown that the second relocation of PKCα occurred during the blastocyst formation. PKCα had different expression patterns in the three kinds of preimplantation embryos. However, the effects of PKCα on embryonic development started in early stage. There must be a necessary connection between PKCα relocation and cell adhesion starting at embryonic compaction.

[Dynamic changes of microtubule in parthenogenetic and in vitro fertilized preimplantation embryos in mouse.].

In this study we detected dynamic changes and function of beta-tubulin, a subtype of microtubule, during the first cleavage period in mouse parthenogenetic and in vitro fertilized embryos. Firstly, we compared the developmental potential of in vitro fertilized, parthenogenetic, and in vivo fertilized embryos in culture. Then, the dynamic changes of beta-tubulin and nucleus in parthenogenetic and in vitro fertilized preimplantation embryos were detected by immunofluorescence and confocal microscopy to analyze the role of microtubules in meiotic division and embryonic development. The results indicated that the development rate of in vivo fertilized embryos was significantly higher than that of in vitro fertilized or parthenogenetic embryos (P<0.05). However, there was no significant difference in developmental potential between in vitro fertilized and parthenogenetic embryos. During in vitro fertilization, oocyte was activated when sperm entered it. Oocyte resumed the second meiotic division. Condensed maternal chromosomes aligning at the equator of the spindle were pulled to the spindle poles by kinetochore microtubules in anaphase. Furthermore, in telophase, there were microtubules between the two sets of decondensed maternal chromosomes. One set formed the second polar body (Pb(2)), which was extruded to the perivitelline space. The other set formed female pronucleus. Meanwhile, 5-8 h after fertilization, sperm chromatin condensed and decondensed to form male pronucleus. Microtubule composed mesosome and cytaster remodeling around male and female pronuclei to form long microtubules, which pull the pronuclei to get close. During 4-6 h parthenogenetic activation, SrCl(2) activated oocytes to resume meiosis. As a consequence, sister chromatids were pulled to spindle poles. Cytochalasin B, which was applied in the medium, inhibited the extrusion of Pb(2). Two haploid pronuclei in the cytoplasm were connected by microtubules. Compared with that in in vitro fertilization, oocyte is easier to be activated in parthenogenetic activation. Chemical activation is more efficient than sperm penetration in in vitro fertilization as indicated by earlier and better remodeling of the microtubules.

Isolation and characterization of a quinclorac-degrading Actinobacteria Streptomyces sp. strain AH-B and its implication on microecology in contaminated soil.

Quinclorac, a highly selective auxin herbicide, is widely used for controlling weeds in rice field. However, the residual quinclorac is toxic to many crops, vegetables, and aquatic animals, resulting in one of the major problems in crop rotation. Here, we investigated the degradation of quinclorac by strain AH-B, which was isolated from long-term quinclorac-contaminated soil using continuous circulating fluidized bed reactor and subjected to atmospheric and room temperature plasma mutation. Morphological examination, 16S rRNA gene sequencing, and phylogenetic analysis revealed that strain AH-B was Streptomyces sp. The quinclorac degradation efficiency of AH-B in liquid medium was 97.2% after 18 days when the initial quinclorac concentration was 20 mg L-1. The degradation products were 3-chloro-7-methoxy-8-quinoline-carboxylic, 3-chloro-7-methyl-8-quinoline-carboxylic, 3-chloro-7-oxyethyl-8-quinoline-carboxylic, and 3,7-dichloro-6-methyl-8-quinoline-carboxylic. The inoculum size, initial quinclorac concentration, pH, and temperature were found to affect quinclorac degradation efficiency of AH-B. High-performance liquid chromatography-electrospray ionization tandem mass spectrometry analysis revealed that quinclorac degradation by AH-B produced many products. In soil with initial quinclorac content of 1 mg kg-1 dry soil, addition of AH-B resulted in 87.5% quinclorac degradation after 42 days, while that in the control (without AH-B) was 22.4%. Furthermore, microecological analysis using next-generation sequencing of 16S rRNA geneshowed that some bacterial species, such as Bacterioides and Proteobacteria, could survive in quinclorac-contaminated soil, while some bacteria, such as Firmicutes, were very sensitive to quinclorac. Besides, some fungal species, such as Basidiomycota, could also survive quinclorac-contamination. After 42 days, the diversity of bacteria and fungi in soil treated with AH-B was higher than that in the control, implying that bioaugmentation with strain AH-B could reduce quinclorac toxicity to microorganisms in soil.