BMP4 Regulates EMT to be Involved in non-Syndromic Cleft lip With or Without Palate.

OBJECTIVE: In the previous study, we identified bone morphogenetic protein 4 (BMP4) responsible for non-syndromic cleft lip with or without cleft palate (NSCL/P). We aimed to elucidate the effects and mechanisms of BMP4 on epithelial-mesenchymal transition (EMT) through Smad1 signaling pathway to be involved in NSCL/P. METHODS: The human oral epidermoid carcinoma cells (KBs) were transfected with plasmids or small interfering RNA (siRNA) to build the models. The migration of the cells was evaluated by transwell assay. Western blotting and quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR) were used to detect the expressions of BMP4, E-cadherin, N-cadherin, EMT-related transcription factors snal1 and snal2, matrix metalloproteinase 2 (MMP2), MMP9, Smad1, and phosphorylated Smad1. RESULTS: In the overexpression group, the migration number of cells was increased significantly. The protein expression of E-cadherin was decreased significantly, while the protein expression level of the N-cadherin was increased significantly. The protein and mRNA expressions of MMP2, MMP9, snal1, and snal2 were significantly higher. The expression level of Smad1 was not significantly changed, while the phosphorylation of Smad1 was significantly increased. In the BMP4-siRNA group, the migrating number cells was significantly decreased. The protein expression of E-cadherin was increased significantly, while the expression of N-cadherin was significantly decreased. The protein and mRNA expressions of MMP2, MMP9, snal1, and snal2 were significantly lower than that of the control group. The expressions of Smad1 and phosphorylation of Smad1 were not significantly changed. CONCLUSIONS: BMP4 enhances cell migration and promotes cell EMT through Smad1 signaling pathway. Abnormal BMP4 mediates migration and EMT through other relevant signaling pathways resulting in NSCL/P. The study provides new insight into the mechanisms of NSCL/P associated with BMP4.n.

Evaluation of conditioned medium from placenta-derived mesenchymal stem cells as a storage medium for avulsed teeth: An in vitro study.

BACKGROUND/AIM: The viability of periodontal ligament cells on the root surface is a major factor that influences the healing of replanted teeth. A suitable storage medium is necessary to preserve avulsed teeth before replantation. Conditioned medium from placenta-derived mesenchymal stem cells (PMSC-CM) contains a variety of growth factors. The aim of this study was to evaluate the effectiveness of PMSC-CM as a storage medium to maintain the cell viability of avulsed teeth. MATERIAL AND METHODS: Extracted premolars from healthy humans were randomly stored in Hank's balanced salt solution (HBSS) and PMSC-CM for 6, 12 and 24 hours, respectively, at room temperature, and then the ratio of apoptosis of the periodontal ligament (PDL) cells was identified by flow cytometry. Human periodontal ligament stem cells (PDLSCs) were incubated with HBSS and PMSC-CM, respectively, for 6, 12, 24 and 48 hours in 5% CO2 at 37°C. Then, the cell viability of PDLSCs was determined using the cell counting kit-8 (CCK-8) and a cell cycle assay was performed. RESULTS: The apoptosis rate of PDL cells in PMSC-CM was significantly lower than that in HBSS at 24 hours (P < .001), while the two groups showed similar cell apoptosis rates at 6 and 12 hours (P > .05). The cell proliferation of PDLSCs treated with PMSC-CM significantly increased compared with the HBSS group (P < .05). The cell cycle assay revealed that the PDLSCs treated with HBSS were arrested at the G1 phase, while there was no difference between the PMSC-CM group and the control group (P > .05). CONCLUSIONS: Compared with HBSS, PMSC-CM showed better inhibition of apoptosis of PDL cells and promoted the proliferation of PDLSCs. Thus, PMSC-CM could be a promising storage medium for avulsed teeth.

[Comparison of platinum-containing doublet chemotherapy and liposome paclitaxel monotherapy for the treatment of elderly patients with advanced non-small cell lung cancer].

OBJECTIVE: Given controversy remains on monotherapy and combinatory chemotherapy in elderly patients (> or = 70 years) with advanced non-small cell lung cancer (NSCLC), we conducted this study to compare the safety and efficacy of liposome paclitaxel and platinum-containing doublets. METHODS: From January 2007 to March 2009, totally 65 patients (age > or = 70 years) with pathologically confirmed NSCLC were enrolled. 33 patients received liposome paclitaxel monotherapy (monotherapy group) and 32 patients received platinum-containing doublets chemotherapy (combinatory group). RESULTS: No CR was observed in all patients. Both groups had similar objective response rate (ORR) (6.1% vs. 15.6%, P = 0.399). However, a statistically significant higher disease control rate (DCR) (65. 6%) was observed in he combinatory group when compared with that of monotherapy group (39.4%, P = 0.034). The combinatory group had longer time-to-progression (TTP) (94 days, 95% CI: 60-127 days) than the monotherapy group (51 days, 95% CI: 22-79 days, P = 0.046). The median overall survival days in the combinatory group was 524 days (95% CI: 146-901 days) where as in the monotherapy group only 146 days (95% CI 32-259 days) (P = 0.001). The most common adverse reactions were myelosuppression, gastrointestinal reactions and elevated transaminase in the monotherapy group, while those were myelosuppression, gastrointestinal reactions and infection in the combinatory group. Generally there was no significant difference in the adverse reaction, except grade 3-4 thrombocytopenia (P = 0.004). It should be addressed that 1 patient (3.0%) in the monotherapy group had an onset of severe infection, while the number rose to 5 (15.6%) in the combinatory group (P = 0.079). CONCLUSION: Platinum-containing doublet chemotherapy achieved a higher response rate, longer time-to-progression and overall survival compared with liposome paclitaxel monotherapy in the treatment of elderly patients with advanced NSCLC. However thrombocytopenia and severe infection should be monitored for the combinatory chemotherapy.

Oral diet management for carcinoma at the base of tongue with radiotherapy and chemotherapy associated dysphagia: a case report.

Introduction: Tongue cancer is one of the common malignancy of the head and neck, and directly impacts chewing, swallowing, and other eating activities. Based on the evidence-based guidelines and clinical management, this paper presents nutrition management experience of a patient with tongue cancer who had a dysphagia and feeding reflux while undergoing radiotherapy and chemotherapy. Methods: Nutritional risk screening and comprehensive nutritional assessment were performed based on the patient's medical history, and personalized nutritional programs were developed under the guidance of the clinical pharmaceutical consensus of parenteral nutrition and nutritional treatment guidelines for patients with tumors during radiotherapy. For the management of oral feeding, the patient's swallowing function was evaluated to manage oral feeding. Thickening powders were used to improve the consistency of the patient's food, which successfully achieved oral feeding of the patient. Results: The patient finally ate five meals a day by mouth, and energy requirements were met using industrialized nutritional supplements, and homogenized food was added in between the meals. The energy provided by enteral nutrition can reached approximately 60-75%. The patient's weight and albumin levels had increased significantly at the time of discharge. Discussion: The nutritional management of patients with dysphagia should be jointly managed by clinicians, nurses, nutritionists, and family members to effectively improve the quality of life (QOL) and nutritional status of patients. To ensure adequate nutritional supply, appropriate swallowing training may delay the deterioration of the chewing function and improve the eating experience of such patients.

A promising cancer gene therapy agent based on the matrix protein of vesicular stomatitis virus.

The matrix (M) protein of vesicular stomatitis virus (VSV) plays a key role in inducing cell apoptosis during infection. To investigate whether M protein-mediated apoptosis could be used in cancer therapy, its cDNA was amplified and cloned into eukaryotic expression vector pcDNA3.1(+). The recombinant plasmid or the control empty plasmid pcDNA3.1(+) was mixed with cationic liposome and introduced into various tumor cell lines in vitro, including lung cancer cell LLC, A549, colon cancer cell CT26 and fibrosarcoma cell MethA. Our data showed that the M protein induced remarkable apoptosis of cancer cells in vitro compared with controls. Fifty micrograms of plasmid in a complex with 250 microg cationic liposome was injected intratumorally into mice bearing LLC or MethA tumor model every 3 days for 6 times. It was found that the tumors treated with M protein plasmid grew much more slowly, and the survival of the mice was significantly prolonged compared with the mice treated with the control plasmid. In MethA fibrosarcoma, the tumors treated with M protein plasmid were even completely regressed, and the mice acquired longtime protection against the same tumor cell in rechallenge experiments. Both apoptotic cells and CD8(+) T cells were widely distributed in M protein plasmid-treated tumor tissue. Activated cytotoxic T lymphocytes (CTLs) were further detected by means of (51)Cr release assay in the spleen of the treated mice. These results showed that M protein of VSV can act as both apoptosis inducer and immune response initiator, which may account for its extraordinary antitumor effect and warrant its further development in cancer gene therapy.

A review of emerging bone tissue engineering via PEG conjugated biodegradable amphiphilic copolymers.

Defects in bones can be caused by a plethora of reasons, such as trauma or illness, and in many cases, it poses challenges to the current treatment approaches for bone repair. With increasing demand of bone bioengineering in tissue transplant, there is a need to source for sustainable solutions to induce bone regeneration. Polymeric biomaterials have been identified as a promising approach due to its excellent biocompatibility and controllable biodegradability. Specifically, poly(ethylene glycol) (PEG) is one of the most commonly investigated polymer for use in bio-related application due to its bioinertness and versatility. Furthermore, the hydrophilic nature enables it to be incorporated with hydrophobic but biodegradable polymers like, polylactide (PLA) and polycaprolactone (PCL), to create an amphiphilic polymer. This article reviews the recent synthetic strategies available for the construction of PEG conjugated polymeric system, analysis of PEG influence on the material properties, and provides an overview of its application in bone engineering.

Vesicular stomatitis virus matrix protein gene enhances the antitumor effects of radiation via induction of apoptosis.

Vesicular stomatitis virus (VSV) matrix (M) protein can directly induce apoptosis by inhibiting host gene expression when it is expressed in the absence of other viral components. Previously, we found that the M protein gene complexed to DOTAP-cholesterol liposome (Lip-MP) can suppress malignant tumor growth in vitro and in vivo; however, little is known regarding the biological effect of Lip-MP combined with radiation. The present study was designed to determine whether Lip-MP could enhance the antitumor activity of radiation. LLC cells treated with a combination of Lip-MP and radiation displayed apparently increased apoptosis compared with those treated with Lip-MP or radiation alone. Mice bearing LLC or Meth A tumors were treated with intratumoral or intravenous injections of Lip-MP and radiation. The combined treatment significantly reduced mean tumor volumes compared with either treatment alone in both tumor models and prolonged the survival time in Meth A tumor models and the intravenous injection group of LLC tumor models. Moreover, the antitumor effects of Lip-MP combined with radiation were greater than their additive effects when compared with the expected effects of the combined treatment in vivo. This study suggests that Lip-MP enhanced the antitumor activity of radiation by increasing the induction of apoptosis.

Antitumour activity of cationic-liposome-conjugated adenovirus containing the CCL19 [chemokine (C-C motif) ligand 19] gene.

CCL19 [chemokine (C-C motif) ligand 19; also known as MIP-3beta (macrophage inflammatory protein-3beta) or ELC (Epstein-Barr-virus-induced molecule 1 ligand chemokine)], one of the immunostimulatory cytokines, chemoattracts both DCs (dendritic cells) and T-lymphocytes. Adenoviral vector is one of the most used gene delivery vectors for cancer therapy because of its high gene-transfection efficiency. However, its wider application is limited, owing to immune responses that reduce transgene expression and decrease the efficacy of repeated administration. We constructed the recombinant replication deficient adenoviral vectors containing the CCL19 gene (Ad-CCL19) and combined them with PEG-PE [poly(ethylene glycol)-phosphatidylethanolamine]-modified cationic liposomes (Ad-CCL19/PEG-PE) for immunotherapy against murine fibrosarcoma. Although there were hardly any therapeutic differences between Ad-CCL19- and Ad-CCL19/PEG-PE-treated mice that were observed at the second administration, the final results demonstrated that Ad-CCL19/PEG-PE-treated mice survived much longer. The antitumour efficacy may be related to the high level of CCL19 after the final administration and lasting expression of IFN-gamma (interferon-gamma) and IL-12 (interleukin-12) in the Ad-CCL19/PEG-PE-treated group, which were measured by reverse-transcription PCR and ELISA. The results demonstrated that PEG-PE-cationic-liposome-conjugated adenovirus could prolong the expression of the therapeutic gene in vivo and may enhance the antitumour efficacy.

[Anti-tumor effects of recombinant adenovirus encoding survivin encapsulated in cationic liposome].

OBJECTIVE: To explore the anti-tumor effects and mechanisms of recombinant adenovirus encoding survivin encapsulated in cationic liposome. METHODS: CT26 tumor model was established in BALB/c mice. Fifty mice were randomly divided into five groups, including the group treated with recombinant adenovirus encoding survivin encapsulated in cationic liposome (Lip+ Ad-sur), the group of recombinant adenovirus encoding survivin (Ad-sur), the group of recombinant adenovirus encoding null encapsulated in cationic liposome (Lip+Ad-null), the group of liposome (Lip), and the group of PBS alone (PBS). Survival rate of mice, tumor volume, and side effects of treatments were observed. Lymphocytes were activated by adenovirus vaccine to kill tumor cells in vitro and in vivo. CTL assay and histological examination were carried out. RESULTS: Immunotherapy with recombinant adenovirus encoding survivin encapsulated in cationic liposome was effective for inducing protective and therapeutic anti-tumor immunity in CT26 tumor model. In the combination therapy group, the tumor growth was inhibited and the tumor volume was significantly smaller when compared with the controls. The survival rate of mice in the combination therapy group at 7 weeks after inoculation of tumor cells was significantly higher than that of the control group. Histologically, the tumor tissue was markedly necrotic and was infiltrated by lymphocytes. 51Cr assay in vitro indicated that the combination therapy group showed higher specific killing activity against CT26 tumor cells than did the control groups, and the T cells were independent of NK cells. CONCLUSION: Immunotherapy with recombinant adenovirus encoding survivin encapsulated in cationic liposome was noted to have a significant anti-tumor effect on CT26 tumor model.

Histologic comparison between platelet-rich plasma and blood clot in regenerative endodontic treatment: an animal study.

INTRODUCTION: In regenerative endodontic treatment (RET) for immature permanent tooth, better treatment results could be obtained by applying platelet-rich plasma (PRP) as the scaffold rather than the blood clot. The goal of this study was to compare the histologic differences between using PRP and blood clot in RET. METHODS: Three 6-month-old beagles each carrying 9 premolars with double root canals were randomly assigned to the PRP group, blood clot group, or negative control group. All experimental teeth suffered apical periodontitis, and RET was performed. In the blood clot group, bleeding was induced from the periapical tissues to fill the canal space. In the PRP group, autologous PRP was injected into each root canal. The animals were sacrificed 3 months later. Histologic sections were stained with hematoxylin-eosin. Statistical analysis was performed by the Fisher exact test, with the significance set at 0.05. RESULTS: With the ingrowth of cellular cementumlike tissues, the canal wall was thickened, and the apical apex was closed in both the PRP and blood clot groups. Cementocytelike cells were present in the newly formed tissues. Meanwhile, no statistical difference was found in both experimental groups for the average percentage of apical closure, new tissue formation, and pulplike tissue formation. Noticeably, a large number of inflammatory cells were present in some root canals in both groups although the postoperative radiograph revealed the disappearance of periapical radiolucency. CONCLUSIONS: PRP application could be an option for clinical cases in which little or no bleeding were found when irritating the apical tissue during RET.

Theoretical and experimental investigation on dissolution and regeneration of cellulose in ionic liquid.

Density functional theory calculations and atoms in molecules theory were performed to investigate the mechanism of cellulose dissolution and regeneration in 1-ethyl-3-methylimidazolium acetate ([emim]Ac), and (1,4)-dimethoxy-ß-D-glucose (Glc) was chosen as the model for cellulose. The theoretical results show that the interaction of [emim]Ac with Glc is stronger than that of Glc with Glc. Further studies indicate that the anion acetate of [emim]Ac forms strong H-bonds with hydroxyl groups of Glc. It is also observed that the H-bonds between [emim]Ac and Glc are weakened or even destroyed by the addition of water. In addition, both the original and regenerated cellulose samples were characterized with FT-IR, XRD, TGA and SEM. The experimental results prove that cellulose can be readily reconstituted from the [emim]Ac-based cellulose solution by the addition of water and the crystalline structure of cellulose is converted to cellulose II from cellulose I in the original cellulose.

Catalytic conversion of cellulose to 5-hydroxymethyl furfural using acidic ionic liquids and co-catalyst.

Efficient catalytic conversion of microcrystalline cellulose (MCC) to 5-hydroxymethyl furfural (HMF), is achieved using acidic ionic liquids (ILs) as the catalysts and metal salts as co-catalysts in the solvent of 1-ethyl-3-methylimidazo-lium acetate ([emim][Ac]). A series of acidic ILs has been synthesized and tested in conversion of MCC to HMF. The effect of reaction conditions, such as reaction time, temperature, catalyst dosage, metal salts, water dosage, Cu(2+) concentration and various acidic ILs are investigated in detail. The results show that CuCl(2) in 1-(4-sulfonic acid) butyl-3-methylimidazolium methyl sulfate ([C(4)SO(3)Hmim][CH(3)SO(3)]), is found to be an efficient catalyst for catalytic conversion of MCC to HMF, and 69.7% yield of HMF is obtained. A mechanism to explain the high activity of CuCl(2) in [C(4)SO(3)Hmim][CH(3)SO(3)] is proposed. To the best of our knowledge, this report first proposes that the Cu(2+) and [C(4)SO(3)Hmim][CH(3)SO(3)] show better catalytic performance in catalytic conversion of MCC to HMF.

[Distribution and species of mercury in water and sediments from Huangpu River].

Levels of total mercury, soluble mercury and particle mercury in water of Huangpu River change greatly, their average values are (0.4 +/- 0.44) ng/mL, (0.27 +/- 0.42)ng/mL, (0.13 +/- 0.10) ng/mL respectively. Mercury in water is mainly in the form of soluble mercury. Average mercury content in sediment of Huangpu River is relative high and up to (204.03 +/- 97.41) ng/g, with a range of 70.52 ng/g - 387.30 ng/g. Mercury content is high in the middle reach of Huangpu River, especially in section of Xidu-Nanpu Bridge, and low in upstream and downstream. Distribution of mercury is hightly related with distribution of industry plants and farming. Locations with high mercury content in sediment are in the downstream of locations with high mercury content in water. Mercury (in sediments) is mainly in the form of residue, exchangeable ions, and humics-bound, seldom is in the form of carbonate-bound. Contrary to residue-bound mercury, exchangeable mercury is low in the middle reach, and high in upstream and downstream. There mainly are residue-bound mercury and little humics-bound mercury, exchangeable mercury, and carbonate-bound mercury in sediment in profile, and the residue-bound mercury increases irregularly with depth. Nearing the Mouth of Yangtze River, mercury in sediment becomes more active.