Development of chitosan/polycaprolactone-thymol Janus films with directional transport and antibacterial properties for meat preservation.

Reducing contamination from percolate is critical to the preservation of foods with high water content, such as pork. This study aims to develop a novel active packaging material for meat preservation by precisely controlled dual-channel one-step electrospinning. Compared to traditional strategies of preparing Janus films, this method allows for greater flexibility and efficiency. The structure and properties of the Janus film are characterized by scanning electron microscopy (SEM), water contact angle (WCA), directional liquid transport investigation, Thymol release and permeation features, and biocompatibility evaluation. Moreover, the Janus film is applied to the packaging of pork with modified atmosphere packaging to demonstrate its practical application prospects in the food active packaging field. The results revealed that the two sides of the film showed completely different wettability, and the change rate of WCA increased with the increase of the scale of hydrophilic fibers. The permeation features of thymol loaded in the film was consistent with the results of antibacterial properties and biocompatibility assessment. Moreover, the Janus film can effectively prolong the shelf life, improve the quality and safety of the pork.

Starch Properties and Morphology of Eight Floury Endosperm Mutants in Rice.

Besides increasing grain yield, improving rice (Oryza sativa L.) quality has been paid more and more attention recently. Cooking and eating quality (CEQ) is an important indicator of rice quality. Since CEQs are quantitative traits and challenging for measurement, efforts have mainly focused on two major genes, Wx and SSIIa. Chalkiness and floury endosperm significantly affect the eating quality of rice, leading to noticeable changes in CEQ. Due to the easily observable phenotype of floury endosperm, cloning single gene mutations that cause floury endosperm and evaluating changes in CEQs indirectly facilitate the exploration of the minor genes controlling CEQ. In this study, eight mutants with different degrees of floury endosperm, generated through ethylmethane sulfonate (EMS) mutagenesis, were analyzed. These mutants exhibited wide variation in starch morphology and CEQs. Particularly, the z2 mutant showed spherical starch granules significantly increased rapid visco analyzer (RVA) indexes and urea swelling, while the z4 mutant displayed extremely sharp starch granules and significantly decreased RVA indexes and urea swelling compared to the wild type. Additionally, these mutants still maintained correlations with certain RVA profiles, suggesting that the genes PUL, which affect these indexes, may not undergo mutation. Cloning these mutated genes in the future, especially in z2 and z4, will enhance the genetic network of rice eating quality and hold significant importance for molecular marker-assisted breeding to improve rice quality.

Impacts of Angelica Polysaccharide on Proliferation and Differentiation of Mesenchymal Stem Cells of Rat Bone Marrow.

In literature, antiosteoporotic effects of Angelica sinensis root have been confirmed, but the impact of Angelica sinensis polysaccharide (ASP) on osteoblastic or adipogenic distinction of BMSCs is limited. This paper aimed to explore the role of ASP on proliferation and differentiation of rat BMSCs. Rat BMSCs were subjected to isolation and identification through flow cytometry. The proliferation of rat BMSCs under ASP was performed by CCK-8 kit. Measures of osteogenesis under different concentrations of ASP were detected by using alizarin red staining for mesenchymal cells differentiation and ALP activity assay to identify ALP activity. Quantitative RT-PCR was selected to identify osteoblastic or adipogenic biomarkers from a genetic perspective. Likewise, we have evaluated measures of indicators of Wnt/ß-catenin signal. ASP significantly promoted the proliferation, increased osteogenesis, and decreased adipogenesis of rat BMSCs within the limit of 20-60 mg/L in a dose-dependent manner but was suppressed at 80 mg/L. The expression of cyclin D1 and ß-catenin showed a considerable rise over the course of ASP induced osteogenesis. Dickkopf 1 (DKK1) suppressed the regulation of rat BMSCs differentiation through the mediation of ASP. We have observed that ASP upregulated the osteogenic but downregulated adipogenic differentiation of BMSCs, and our findings help to contribute to effective solutions for treating bone disorders.

Intensification of sugar production by using Tween 80 to enhance metal-salt catalyzed pretreatment and enzymatic hydrolysis of sugarcane bagasse.

In this study, different metal-salt catalyzed pretreatment was presented to disorganize the obstinate structure by eliminating the majority of hemicellulose, fractional of lignin, and improve the enzymatic saccharification of sugarcane bagasse. With the accession of Tween 80 during enzymolysis, all metal-salt pretreated substrates presented higher glucose yields, especially for CuCl2. Furthermore, Tween 80 was added to the pretreatment, enhancing the elimination of hemicellulose and lignin, decreasing the degradation of sugars to inhibitors, and presenting superior performance on improving glucose yield. In addition, the maximum glucose yield of 88.0% was achieved by using Tween 80 concomitantly with AlCl3 pretreatment and enzymolysis. It was also found that adding Tween 80 during pretreatment or/and enzymolysis after 24 h could liberate the similar glucose without Tween 80 after 72 h. However, the enhancement of Tween 80 at 6 h was higher than that at 72 h.

CD44-Targeted Magnetic Nanoparticles Kill Head And Neck Squamous Cell Carcinoma Stem Cells In An Alternating Magnetic Field.

BACKGROUND: Head and neck squamous cell carcinoma (HNSCC) is the sixth most common malignant tumor in the world. Studies in recent years have demonstrated that cancer stem cells (CSCs) are present in many tumor tissues, including HNSCC, and CSCs are the root cause of tumor recurrence and metastasis. Thus, taking new treatment measures to target the killing of CSCs that are resistant to chemotherapy and radiotherapy is key to the success of cancer treatment. METHODS: We explored a method for preparing anti-CD44 antibody-modified superparamagnetic iron oxide nanoparticles (SPIONPs). Biocompatibility was evaluated by a CCK-8 assay. The CSCs were obtained by a 3D cell culture technique from Cal-27 (human oral squamous cell carcinoma) cells, and then the CSCs were identified by quantitative real-time polymerase chain reaction (qRT-PCR). The targeting efficiency of the CD44-SPIONPs to CSCs was confirmed by Prussian blue staining and visualized by laser scanning confocal microscopy (LSCM). Flow cytometry was used to detect the apoptosis of CSCs after alternating magnetic field (AMF) treatment. The efficacy of tumor growth inhibition by CD44-SPIONP-mediated magnetic hyperthermia therapy was evaluated with tumor xenografts in nude mice. RESULTS: The CD44-SPIONPs exhibited no negative effect on CSCs, indicating good biocompatibility. After SPIONPs were cocultured with stem cells, the majority of CD44-SPIONPs labeled with FITC penetrated the cell membrane into the cytoplasm. After AMF treatment, CD44-SPIONPs induced CSCs to undergo programmed death. The inhibitory ratio of the treated group was 33.43%, and necrotic areas in the tumor tissue were mainly distributed around the magnetic fluid. CONCLUSION: These results demonstrate that it is possible to kill CSCs using targeted magnetic nanoparticles and an AMF and that magnetic fluid hyperthermia significantly inhibited the growth of grafted Cal-27 tumors in mice.

[Effect of hypoglycosylated E-cadherins on proliferation and invasiveness of tongue squamous cell carcinoma].

PURPOSE: To investigate the effect of N-glycosylated E-cadherins on proliferation and invasiveness of tongue squamous cell carcinoma CAL 27 cells, and to study the effect of hypoglycosylation of E-cadherins on the stability of adherens junctions (AJs) mediated by E-cadherins. METHODS: Human tongue squamous cell carcinoma CAL 27 cells were respectively transfected with the plasmids encoding either hypoglycosylated or wild-type E-cadherins with FLAG as tags. Western blot of FLAG was used to detect the expression of exogenous E-cadherins in the transfected cells after 48 hours. The cell proliferation counting, monolayer cell scratch wound healing and in vitro cell invasiveness assays were performed to evaluate the proliferation and invasiveness of cells. Immunoprecipitation experiments were used to assay the quantity of α-catenins, ß-catenins, γ-catenins and vinculins linked with the cytosolic tails of E-cadherins. All statistical analysis were performed using SPSS 17.0 software package. RESULTS: The expression of exogenous E-cadherins was detected in transfected CAL 27 cells by Western blot. Cell proliferation counting assay revealed that hypoglycosylated E-cadherins significantly inhibited proliferation of CAL 27 cells, compared with wild-type E-cadherins. The monolayer cell scratch wound healing and in vitro cell invasiveness assays indicated that hypoglycosylated E-cadherins also significantly restrained both invasiveness of CAL 27 cells, compared with wild-type E-cadherins. The immunoprecipitation experiments demonstrated that hypoglycosylated E-cadherins exhibited an increased association with α-catenins, ß-catenins, and γ-catenins and vinculins. CONCLUSIONS: Compared with wild-type E-cadherins, hypoglycosylated E-cadherins can significantly suppress proliferation and invasiveness of tongue squamous cell carcinoma CAL 27 cells, and the mechanism was that hypoglycosylated E-cadherins can induce more stable AJs than wild-type E-cadherin. Supported by Scientific Research Foundation for the Returned Oversea Chinese Scholars, State Ministry of Education of China and Shandong Provincial Award Foundation for Excellent Middle- and Young-Aged Scientists (BS2010YY021).

Aberrant amplification of the crosstalk between canonical Wnt signaling and N-glycosylation gene DPAGT1 promotes oral cancer.

Oral cancer is one of the most aggressive epithelial malignancies, whose incidence is on the rise. Previous studies have shown that in a subset of human oral squamous cell carcinoma (OSCC) tumor specimens, overexpression of the DPAGT1 gene, encoding the dolichol-P-dependent N-acetylglucoseamine-1-phosphate transferase, a key regulator of the metabolic pathway of protein N-glycosylation, drives tumor cell discohesion by inhibiting E-cadherin adhesive function. Recently, we reported that DPAGT1 was a target of the canonical Wnt signaling pathway. Here, we link overexpression of DPAGT1 in human OSCC tumor specimens to aberrant activation of canonical Wnt signaling. We report dramatic increases in ß- and γ-catenins at the DPAGT1 promoter and correlate them with reduced expression of a Wnt inhibitor, Dickkopf-1 (Dkk-1). Using human squamous carcinoma cell lines of the head and neck, we show that partial inhibition of DPAGT1 reduces canonical Wnt signaling, indicating that DPAGT1 and canonical Wnt signaling function in a positive feedback loop. We provide evidence that E-cadherin inhibits DPAGT1, canonical Wnt signaling and the OSCC cancer phenotype by depleting nuclear ß- and γ-catenins, with hypoglycosylated E-cadherin being the most effective. This suggests that in human OSCC, extensive N-glycosylation of E-cadherin compromises its ability to inhibit canonical Wnt signaling and DPAGT1 expression. Our studies reveal a novel interplay between DPAGT1/N-glycosylation and canonical Wnt signaling and suggest that dysregulation of this crosstalk is a key mechanism underlying OSCC. They also suggest that partial inhibition of DPAGT1 may represent an effective way to restore normal interactions among these essential pathways in oral cancer.

N-glycosylation status of E-cadherin controls cytoskeletal dynamics through the organization of distinct ß-catenin- and γ-catenin-containing AJs.

N-glycosylation of E-cadherin has been shown to inhibit cell-cell adhesion. Specifically, our recent studies have provided evidence that the reduction of E-cadherin N-glycosylation promoted the recruitment of stabilizing components, vinculin and serine/threonine protein phosphatase 2A (PP2A), to adherens junctions (AJs) and enhanced the association of AJs with the actin cytoskeleton. Here, we examined the details of how N-glycosylation of E-cadherin affected the molecular organization of AJs and their cytoskeletal interactions. Using the hypoglycosylated E-cadherin variant, V13, we show that V13/ß-catenin complexes preferentially interacted with PP2A and with the microtubule motor protein dynein. This correlated with dephosphorylation of the microtubule-associated protein tau, suggesting that increased association of PP2A with V13-containing AJs promoted their tethering to microtubules. On the other hand V13/γ-catenin complexes associated more with vinculin, suggesting that they mediated the interaction of AJs with the actin cytoskeleton. N-glycosylation driven changes in the molecular organization of AJs were physiologically significant because transfection of V13 into A253 cancer cells, lacking both mature AJs and tight junctions (TJs), promoted the formation of stable AJs and enhanced the function of TJs to a greater extent than wild-type E-cadherin. These studies provide the first mechanistic insights into how N-glycosylation of E-cadherin drives changes in AJ composition through the assembly of distinct ß-catenin- and γ-catenin-containing scaffolds that impact the interaction with different cytoskeletal components.

Combined effects of soluble vascular endothelial growth factor receptor FLT-1 gene therapy and cisplatin chemotherapy in human tongue carcinoma xenografts.

The aim of the present study was to assess the anti-tumor effect of a defective adenovirus that expresses soluble vascular endothelial growth factor (VEGF) receptor FLT-1 (AdsFLT-1) in combination with cisplatin (cis-diamminedichloroplatinum, DDP) on human tongue carcinoma Tca8113 cell xenografts that had been pre-established in nude mice. In vitro, Tca8113 cells secreted soluble FLT-1 (sFLT-1) after infection with AdsFLT-1, and the conditioned medium from AdsFLT-1-treated Tca8113 cells seemed to inhibit VEGF-induced proliferation of human umbilical vein endothelial cells. The combined effects of sFLT-1 gene therapy and DDP chemotherapy was then studied in well-established Tca8113 xenografts. The concentration of sFLT-1 in serum reached a peak 8 days after intratumoral injection of AdsFLT-1. In these tumors, AdsFLT-1 intratumoral injections had only a small effect. Interestingly, when the cells were also exposed to DDP chemotherapy, significantly higher (P<0.05), and possibly synergistic, anti-tumoral effects were observed that were highly correlated to a marked reduction in intratumoral vascularization and an increase in tumor-cell apoptosis. Together, these data emphasize the potential of combining an anti-angiogenic gene therapy strategy with a destructive approach directed against the tumor cells to fight human tongue carcinoma.

[Clinical application of maxillary sinus lifting, bone graft and simultaneous placement of implant].

PURPOSE: To evaluate the results of two kinds of sinus lifting techniques with simultaneous implant placement. METHODS: 31 maxillary sinus underwent two kinds of sinus lifting techniques and simultaneously 42 implants placement. The sinuses were observed 1, 3, 6 months after the surgery. RESULTS: There were no implants loose or lost; X ray examination showed well osseointegration and no maxillary sinusitis. All the patients finished implant prosthesis in 6 months postoperatively. Through 6-36 months' follow-up, clinical results were satisfactory. CONCLUSION: With properly handling of indication and the operation skill, the results of two kinds of maxillary sinus lifting techniques with bone grafting and simultaneously implants placement are both satisfactory.

[Expression and secretion of human tumor necrosis factor gene transfected on human embryo myoblasts].

OBJECTIVE: To observe human tumor necrosis factor-alpha (hTNF-alpha) expression and secreting level of human embryo myoblasts transfected by hTNF-alpha gene. METHODS: Human embryo myoblasts were transfected with shuttle plasmid pSV23SHTNF containing hTNF-alpha gene by cationic liposomes DOSPER. The control group was only given equivalent liposomes except plasmid. After culturing for 24, 48, 72 and 96 hours, hTNF-alpha expression level of human embryo myoblasts was observed with immunocytochemistry staining, and hTNF-alpha secreting of human embryo myoblasts was analyzed by ELISA. RESULTS: After transfected by hTNF-alpha gene for 24, 48, 72 and 96 hours, the human embryo myoblasts displayed significant secretion of hTNF-alpha in the cultural supernatant (P < 0.05), and overexpression in cytoplasma and cell membrane. CONCLUSION: Transfection of hTNF-alpha gene to human myoblasts made myoblasts secrete high concentration of hTNF-alpha, implying it is feasible that transfecting muscle cells surrounding tongue carcinoma lesion with hTNF-alpha gene can prevent tongue carcinoma from intruding into deeper muscle tissue.

Induction of apoptosis and tumor regression by vesicular stomatitis virus in the presence of gemcitabine in lung cancer.

Vesicular stomatitis virus (VSV) has been shown to replicate rapidly in vitro and kill selectively a variety of tumor cell lines. The present study was designed to determine whether gemcitabine potentiates the antitumor activity of VSV in vitro and in vivo. A549 human lung adenocarcinoma cells and LLC Lewis lung carcinoma cells were treated with VSV (0.1-10 plaque-forming units per cell) plus gemcitabine (20 nM to 20 microM). Mice bearing A549 or LLC were treated with VSV (5 x 10(4) to 1 x 10(8) plaque-forming units) daily for 5 days plus gemcitabine (5-125 mg/kg/day) once every 3 days for 4 times. Induction of apoptosis and effects on growth inhibition were assessed. The lung cancer cells treated with VSV plus gemcitabine displayed the apparently increased apoptotic cells compared with treatment with VSV or gemcitabine alone. The combined treatment with VSV plus gemcitabine induced the apparent antitumor activity with complete regression of the established lung cancer in both A549 and LLC lung cancer models and augmented the induction of apoptosis in lung cancer cells in vivo as well. This study suggests that the combined treatment with VSV plus gemcitabine may augment the induction of apoptosis in lung cancer cells in vitro and in vivo, and that the augmented antitumor activity in vivo may result from the increased induction of apoptosis in lung cancer cells. The present findings may be of importance to the further exploration of the potential application of this combined approach in the treatment of lung cancer.

[Suppression effect of human tumor necrosis factor-alpha gene transfection on tongue carcinoma cells].

OBJECTIVE: The aim of this study was to investigate suppression effects of the transfection of human tumor necrosis factor-alpha (hTNF-alpha) gene on tongue carcinoma cells. METHODS: The shuttle plasmid containing hTNF-alpha gene was extracted and purified, then it was transferred into Tca8113 tongue carcinoma cells with cationic liposome DOSPER. The control group was only given equivalent liposomes, except the plasmid. After culturing for 24, 48, 72 and 96 hours, the expression of hTNF-alpha gene in Tca8113 cells was analysed by ELISA and, the survival rate of transferred cells was assayed by MTT enzymatic labeling technique. RESULTS: The transferred Tca8113 cells displayed significantly overexpression of hTNF-alpha (P < 0.05). The survival rate of the transferred Tca8113 cells was decreased significantly (P < 0.05). CONCLUSION: Transfection of hTNF-alpha gene in vitro mediated by cationic liposomes can induce the overexpression of hTNF-alpha and inhibit the growth of tongue carcinoma cells.

[Synergistic effects of human tumor necrosis factor-alpha gene transfection and interferon-gamma on the growth of tongue carcinoma cells].

OBJECTIVE: The purpose of this study is to investigate the synergistic effects of human tumor necrosis factor-alpha (hTNF-alpha) transfection and interferon-gamma (IFN-gamma) on the growth of tongue carcinoma cells. METHODS: The cultured Tca8113 tongue carcinoma cells was divided into 2 groups, one group was transferred with hTNF-alpha gene. Each of the 2 groups was then divided into 5 subgroups, and the subgroups were added IFN-gamma until the final IFN-gamma concentrations respectively were 0, 10, 100, and 1000 U/ml. After culturing for 48 hours, the survival rates of the all groups of cells were assayed by MTT enzymatic labeling technique, and the expression of hTNF-alpha in Tca8113 cells was observed with immunocytochemistry. RESULTS: IFN-gamma did not affect the growth of Tca8113 cells without hTNF-alpha, however, the transfection of hTNF-alpha with the above different concentrations of IFN-gamma synergistically inhibited the growth of Tca8113 cells, the concentrations of IFN-gamma were positively correlated with the inhibition effects (r = 0.733, P < 0.01), the transferred Tca8113 cells displayed remarkable overexpression of hTNF-alpha, compared with the non-transferred. CONCLUSION: IFN-gamma can enhance the inhibition of hTNF-alpha transfection on the tongue carcinoma cells.