CCL17 combined with CCL19 as a nasal adjuvant enhances the immunogenicity of an anti-caries DNA vaccine in rodents.

AIM: CCL19 and its receptor CCR7 are essential molecules for facilitating the trafficking of mature dendritic cells (DCs) and helping to establish a microenvironment in lymphoid tissues to initiate primary immune responses, whereas CCL17 is required in the CCR7-CCL19-dependent migration of DCs. In this study we examined whether co-administration of CCL17 and CCL19 could enhance the immunogenicity of an anti-caries DNA vaccine, pCIA-P, in rodents. METHODS: Plasmids encoding CCL17 (pCCL17/VAX) and CCL19 (pCCL19/VAX) were constructed. BALB/c mice were intranasally administered pCCL17/VAX, pCCL19/VAX, or pCCL17/VAX plus pCCL19/VAX, the migration of DCs to the spleen and draining lymph nodes (DLNs) was assessed with flow cytometry. The mice were co-administered pCIA-P; and the anti-PAc antibodies in the serum and saliva were detected with ELISA. Wistar rats were orally challenged with Streptococcus mutans and then administered pCIA-P in combination with pCCL17/VAX, pCCL19/VAX, or pCCL17/VAX plus pCCL19/VAX. The amount of S mutans sustained on rat molar surfaces was assessed using a colony forming assay. Caries activity was scored with the Keyes method. RESULTS: Co-administration of the CCL17 and CCL19 genes in mice caused a greater increase in the number of mature DCs in the spleen and DLNs compared with administration of CCL17 or CCL19 genes alone. CCL17 and CCL19 double-adjuvant plus pCIA-P induced significantly higher levels of anti-PAc salivary IgA and anti-PAc serum IgG antibody in mice, and strengthened the ability of pCIA-P in inhibiting the colonization of S mutans on rat tooth surfaces. The caries activity of the combined adjuvant group was significantly lower than that of the pCCL17/VAX or the pCCL19/VAX group. CONCLUSION: A nasal adjuvant consisting of a combination of CCL17 and CCL19 attracts more mature DCs to secondary lymphoid tissues, inducing enhanced antibody responses against the anti-caries DNA vaccine pCIA-P and reducing S mutans infection in rodents.

Copy-number mutations on chromosome 17q24.2-q24.3 in congenital generalized hypertrichosis terminalis with or without gingival hyperplasia.

Congenital generalized hypertrichosis terminalis (CGHT) is a rare condition characterized by universal excessive growth of pigmented terminal hairs and often accompanied with gingival hyperplasia. In the present study, we describe three Han Chinese families with autosomal-dominant CGHT and a sporadic case with extreme CGHT and gingival hyperplasia. We first did a genome-wide linkage scan in a large four-generation family. Our parametric multipoint linkage analysis revealed a genetic locus for CGHT on chromosome 17q24.2-q24.3. Further two-point linkage and haplotyping with microsatellite markers from the same chromosome region confirmed the genetic mapping and showed in all the families a microdeletion within the critical region that was present in all affected individuals but not in unaffected family members. We then carried out copy-number analysis with the Affymetrix Genome-Wide Human SNP Array 6.0 and detected genomic microdeletions of different sizes and with different breakpoints in the three families. We validated these microdeletions by real-time quantitative PCR and confirmed their perfect cosegregation with the disease phenotype in the three families. In the sporadic case, however, we found a de novo microduplication. Two-color interphase FISH analysis demonstrated that the duplication was inverted. These copy-number variations (CNVs) shared a common genomic region in which CNV is not reported in the public database and was not detected in our 434 unrelated Han Chinese normal controls. Thus, pathogenic copy-number mutations on 17q24.2-q24.3 are responsible for CGHT with or without gingival hyperplasia. Our work identifies CGHT as a genomic disorder.

Intertwined carbon networks derived from Polyimide/Cellulose composite as porous electrode for symmetrical supercapacitor.

Designing intertwined porous structure is highly desirable to improve the electrochemical performance of carbon materials for supercapacitor. In this contribution, three-dimensional (3D) carbonized polyimide/cellulose (CPC) composite is fabricated via a facile "one-step" carbonization, in which cellulose as cross-linked agent is capable of modulating the molecular structure of polyamic acid, thus ensuring the formation of intertwined porous networks in the obtained carbon skeleton. Benefitting from the high specific surface area (951 m2 g-1) and uniformly distributed pores, the optimized CPC-5 electrode exhibits an outstanding specific capacitance of 300F g-1 in 6.0 M KOH electrolyte. More impressively, the CPC-5 based symmetrical supercapacitor affords a high energy density of 22.6 Wh kg-1 at power density of 800 W kg-1, as well as an exceptional capacitance retention of 91.4% after 10,000 cycles. This work affords an effective strategy to yield a promising polyimide derived carbon material for high-performance supercapacitors.

Linc02349 promotes osteogenesis of human umbilical cord-derived stem cells by acting as a competing endogenous RNA for miR-25-3p and miR-33b-5p.

OBJECTIVES: Increasing evidences suggest that inducing mesenchymal stem cells to differentiate into osteoblasts has been as an especially important component in the prevention and therapy for degenerative bone disease. Here, we identify a novel lncRNA, linc02349, which increases significantly during osteogenic differentiation. MATERIALS AND METHODS: Human umbilical cord-derived stem cells (hUC-MSCs) and dental pulp mesenchymal stem cells were used. Overexpression and knockdown of linc02349 in cell lines were generated using lentiviral-mediated gene delivery method. Bioinformatics prediction, Ago2-RIP assay and dual-luciferase reporter system were employed to examine miRNA which interacts with linc02349. The RNA FISH assay was performed to identify the subcelluar location of linc02349. Alizarin Red S staining, ALP staining and qPCR were applied to identify the osteogenic differentiation. The potential linc02349-regulated genes, miR-25-3p and miR-33b-5p, were explored by ChIP, RIP and Western blotting assays. Micro-CT was used to measure the osteogenic content in bone formation assay in vivo. RESULTS: Linc02349 overexpression improves osteogenic differentiation by in vitro and in vivo analysis. Mechanistically, linc02349 acts as a molecular sponge for miR-25-3p and miR-33b-5p to control expression abundance of SMAD5 and Wnt10b, respectively, which eventually activated Dlx5/OSX pathway and hence promoted osteogenic differentiation. In addition, we revealed that STAT3 interacts with linc02349 promoter region and positively regulates the linc02349 transcriptional activity. CONCLUSION: These findings identify that linc02349 modulates the osteogenic differentiation through acting as a sponge RNA of miR-25-3p and miR-33b-5p and regulating SMAD5 and Wnt10b, and proposed a new interaction between STAT3 and linc02349, which could be a potential target in the process the osteogenesis of hUC-MSCs for future clinical application.

The exploration of endocytic mechanisms of PLA-PEG nanoparticles prepared by coaxialtri-capillary electrospray-template removal method.

The nano-sized poly(lactic acid)-poly(ethylene glycol) (PLA-PEG) particles with core-shell structure were efficiently prepared by using coaxial tri-capillary electrospray-template removal method. The cellular uptake mechanism, intracellular distribution and exocytosis in A549 cell model of electrosprayed PLA-PEG nanoparticles were systemically studied. The drug release behavior of electrosprayed PLA-PEG nanoparticles were also investigated. Our results showed that PLA-PEG nanoparticles can be endocytosed quickly by A549 cells. The cellular uptake of PLA-PEG nanoparticles was an energy dependent endocytosis process. Caveolae-mediated endocytosis was only one of endocytosis pathways in A549 cells for PLA-PEG nanoparticles, while clathrin mediated endocytosis was not involved in the endocytosis process. The endocytosed PLA-PEG nanoparticles enriched in the head of A549 cells and only a small amount of them was transported into lysosome after 24h incubation. These findings provided insights into the application of electrosprayed PLA-PEG nanoparticles in nano drug delivery field.

Effects of 1,25-dihydroxyvitamin D3 on Macrophage Cytokine Secretion Stimulated by Porphyromonas gingivalis.

Vitamin D is known to be closely associated with periodontitis; however, its exact mechanisms remain to be clarified. The present study aimed to investigate the influence of 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) on Porphyromonas gingivalis (Pg)-stimulated cytokine production and the involved signaling pathways in macrophages. The main observation was that 1,25(OH)2D3 inhibited Pg-induced interleukin (IL)-6 cytokine expression but up-regulated the expression of anti-inflammatory cytokine IL-10. Further analyses showed that 1,25(OH)2D3 decreased p38 mitogen-activated protein kinase (MAPK) and extracellular signal regulated kinase (ERK)1/2 phosphorylation. Inhibited phosphorylation of p38 MAPK and ERK1/2 was associated with decreased level of IL-6 expression, but was not related to increased level of IL-10 expression in macrophages stimulated with Pg. These results suggest that 1,25(OH)2D3 might exert its anti-inflammatory effects on Pg-stimulated macrophages partly through its inhibitory effect on the p38 MAPK and ERK1/2 signaling pathway.

Inhibitory effect of 1,25-dihydroxyvitamin D3 on Porphyromonas gingivalis-induced inflammation and bone resorption in vivo.

OBJECTIVE: To investigate whether intragastric administration of 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) could inhibit the bone resorption and inflammation in a mouse calvarial model infected by Porphyromonas gingivalis (P. gingivalis). DESIGN: Live P. gingivalis ATCC 33277 was injected once daily for 6days into the subcutaneous tissue overlying the calvaria in mice. At the same time, 1,25(OH)2D3 (50µg/kg per day) was administered by gavage for 9days, starting 3d before the infection. Mice were killed under ether anesthesia 8h after the last injection of P. gingivalis. Micro-computed tomography scanning was used to evaluate calvarial bone loss. Tartrate-resistant acid phosphatase was used to detect osteoclast activity. Real-time PCR was used to assess the mRNA expressions of OPG, RANKL, c-Fos, NFATc1, CTSK and TRAP in calvarial bone and IL-6, IL-10, IL-1ß, IL-12p40 and TNF-α in soft tissue. The levels of serum IL-6, IL-10 were determined by ELISA. RESULTS: 1,25(OH)2D3 treatment apparently attenuated bone resorption in P. gingivalis-induced mouse calvarial model and markedly reduced the number of osteoclasts. The expression levels of RANKL and osteoclast-related genes such as c-Fos, NFATc1, CTSK and TRAP were also decreased by 1,25(OH)2D3. Besides, 1,25(OH)2D3 inhibited the expression of pro-inflammatory cytokines IL-6, IL-12p40 and TNF-α and enormously elevated the expression of anti-inflammatory cytokine IL-10. CONCLUSION: 1,25(OH)2D3 may decrease bone resorption in vivo via suppressing the expression of osteoclast-related genes and its anti-inflammatory properties.

Direct Pulp Capping with Calcium Hydroxide or Mineral Trioxide Aggregate: A Meta-analysis.

INTRODUCTION: The purpose of this study was to compare the effectiveness of mineral trioxide aggregate (MTA) and calcium hydroxide (CH) as pulp capping materials in humans by means of a meta-analysis. METHODS: The PubMed, Cochrane Library, Embase, and Web of Knowledge databases were used in the literature search from their establishment date until December 7, 2014. Studies that met the inclusion criteria were accepted, and necessary information was extracted by 2 authors independently using a standardized form. The success rate, inflammatory response, and dentin bridge formation were evaluated. RESULTS: Thirteen studies met the inclusion criteria. There was no significant heterogeneity between studies, so a fixed-effects model was used. The MTA treatment groups showed a significantly higher success rate compared with CH-capped groups (randomized controlled trials: odds ratio [OR] = 2.26; 95% confidence interval [CI] = 1.33-3.85; P = .003; retrospective nonrandomized trials: OR = 2.88; 95% CI, 1.86-4.44; P < .00001). MTA was superior to CH in terms of the absence of an inflammatory response as well as dentin bridge formation, with the OR being 4.56 (95% CI, 2.65-7.83) and 3.56 (95% CI, 1.89-6.70), respectively. CONCLUSIONS: MTA has a higher success rate and results in less pulpal inflammatory response and more predictable hard dentin bridge formation than CH. MTA appears to be a suitable replacement of CH used for direct pulp capping.

Generation of nano-sized core-shell particles using a coaxial tri-capillary electrospray-template removal method.

This study proposed a new strategy based on a coaxial tri-capillary electrospray-template removal process for producing nanosized polylactide-b-polyethylene glycol (PLA-PEG) particles with a core-shell structure. Microparticles with core-shell-corona structures were first fabricated by coaxial tri-capillary electrospray, and core-shell nanoparticles less than 200 nm in size were subsequently obtained by removing the PEG template from the core-shell-corona microparticles. The nanoparticle size could be modulated by adjusting the flow rate of corona fluid, and nanoparticles with an average diameter of 106±5 nm were obtained. The nanoparticles displayed excellent dispersion stability in aqueous media and very low cytotoxicity. Paclitaxel was used as a model drug to be incorporated into the core section of the nanoparticles. A drug loading content in the nanoparticles as high as 50.7±1.5 wt% with an encapsulation efficiency of greater than 70% could be achieved by simply increasing the feed rate of the drug solution. Paclitaxel exhibited sustained release from the nanoparticles for more than 40 days. The location of the paclitaxel in the nanoparticles, i.e., in the core or shell layer, did not have a significant effect on its release.

[A exploration and study of the relationships of hand-foot-mouth disease (HFMD) and the climate].

OBJECTIVE: To explore and study the relationships between the popularity of HFMD and the climate in Qinhuangdao city. METHODS: HFMD cases were collected on a ten-day basis in 2009 in Qinhuangdao city. At the same time, the data about Qinhuangdao's ten-day average temperature and average humidity were provided by the Qinhuangdao Meteorological bureau. Then the collected data were analysed using the great data analysis function in the EXCELE software. RESULTS: The results showed that the disease of HFMD had a positive relationship with seasons. The cases of HFMD began to rise at the last ten days of March and rised dramatically at the middle ten days of April; In July, the cases of HFMD arrived at peak and then decline gradually. The cases of HFMD in October were quite similar to the cases of HFMD in March. Then in November, the cases of HFMD declined rapidly. All these evidences suggested that the peak seasons of HFMD were Spring and Summer. CONCLUSION: The situations of HFMD had a significant positive relationship with the conditions of climate, such as high temperature and high humidity.

[Distribution of suspended matter concentration in the Changjiang Estuary and adjacent area after the first-stage storage of the Three Gorges Project].

The first-stage storage of the Three Gorges Project started on the 1st June, 2003 and ended on the 10th June, 2003. In order to investigate the influences of the Three Gorges Project on the Changjiang Estuary and adjacent area environment, a cruise was carried out from the 15th June, 2003 to the 25th June, 2003 to synthetically study the environment. Suspended matter concentrations (SMC) in different water layers at different sites were calculated and distributions of SMC were discussed combining with salinity and temperature of water based on recovered data. In results, SMC increase from surface to bottom in Changjiang Estuary and adjacent area. Suspended matters are enriched in the water over the river mouth bar and alongshore area which are located at the west of 123 degrees E. Compared to that before the storage, SMC are reduced obviously in the Changjiang course and south branch after the storage. For example, at the Xuliujing, SMC are reduced from 400 - 500 mg/L to 60.2 mg/L, and in the south branch, from 445 mg/L to 148 mg/L. In other areas, SMC do not show a significant change that they are both less than 10 mg/L in the surface player and 100 mg/L in bottom player. Generally, the distributed patterns of SMC in the investigation area after the storage are still similar to that before the storage.