Comprehensive analysis of M2 macrophage-derived exosomes facilitating osteogenic differentiation of human periodontal ligament stem cells.

BACKGROUND: The role of periodontal ligament stem cells (PDLSCs) and macrophage polarization in periodontal tissue regeneration and bone remodeling during orthodontic tooth movement (OTM) has been well documented. Nevertheless, the interactions between macrophages and PDLSCs in OTM remain to be investigated. Consequently, the present study was proposed to explore the effect of different polarization states of macrophages on the osteogenic differentiation of PDLSCs. METHODS: After M0, M1 and M2 macrophage-derived exosomes (M0-exo, M1-exo and M2-exo) treatment of primary cultured human PDLSCs, respectively, mineralized nodules were observed by Alizarin red S staining, and the expression of ALP and OCN mRNA and protein were detected by RT-qPCR and Western blotting, correspondingly. Identification of differentially expressed microRNAs (DE-miRNA) in M0-exo and M2-exo by miRNA microarray, and GO and KEGG enrichment analysis of DE-miRNA targets, and construction of protein-protein interaction networks. RESULTS: M2-exo augmented mineralized nodule formation and upregulated ALP and OCN expression in PDLSCs, while M0-exo had no significant effect. Compared to M0-exo, a total of 52 DE-miRNAs were identified in M2-exo. The expression of hsa-miR-6507-3p, hsa-miR-4731-3p, hsa-miR-4728-3p, hsa-miR-3614-5p and hsa-miR-6785-3p was significantly down-regulated, and the expression of hsa-miR-6085, hsa-miR-4800-5p, hsa-miR-4778-5p, hsa-miR-6780b-5p and hsa-miR-1227-5p was significantly up-regulated in M2-exo compared to M0-exo. GO and KEGG enrichment analysis revealed that the downstream targets of DE-miRNAs were mainly involved in the differentiation and migration of multiple cells. CONCLUSIONS: In summary, we have indicated for the first time that M2-exo can promote osteogenic differentiation of human PDLSCs, and have revealed the functions and pathways involved in the DE-miRNAs of M0-exo and M2-exo and their downstream targets.

Periodontal disease and risk of mortality and kidney function decline in advanced chronic kidney disease: a nationwide population-based cohort study.

OBJECTIVES: Periodontal disease is prevalent in patients with chronic kidney disease (CKD) and potentially associated with kidney function decline. However, it is uncertain whether periodontal disease affects the risk of mortality and morbidity in patients with advanced CKD. MATERIALS AND METHODS: Taiwan's National Health Insurance Research Database was used to conduct a nationwide population-based cohort study. Propensity score matching procedures were performed to select people with stage 5 CKD and to compare the long-term risk of mortality, end-stage renal disease, and major adverse cardiovascular events (MACE) between people with and without periodontal disease. Multivariable Cox regression analyses were conducted to calculate the adjusted hazard ratio (aHR) with 95% confidence interval (CI) for the outcome of interest. RESULTS: A total of 8119 subjects with stage 5 CKD were initially included. After matching to demographic and clinical covariates, 1254 subjects with 7099 person-years of follow-up were selected for analyses. Periodontal disease was not associated with long-term risks of all-cause mortality (aHR: 0.77, 95% CI: 0.49-1.22), progression to end-stage renal disease (aHR: 0.91, 95% CI: 0.75-1.10), or MACE (aHR: 1.18, 95% CI: 0.91-1.53). These findings were generally consistent across subgroups of age, sex, comorbid diabetes, uses of systemic antibiotic, and different dental procedures. CONCLUSIONS: Periodontal disease is not a predictor for long-term mortality or morbidity in patients with advanced CKD. CLINICAL RELEVANCE: These results provide important evidence to elucidate the relationship between periodontitis and critical clinical outcomes of advanced CKD.

Epidemiology and HBGA-susceptibility investigation of a G9P[8] rotavirus outbreak in a school in Lechang, China.

Rotaviruses cause severe gastroenteritis in infants, in which the viruses interact with human histo-blood group antigens (HBGAs) as attachment and host susceptibility factors. While gastroenteritis outbreaks caused by rotaviruses are uncommon in adolescents, we reported here one that occurred in a middle school in China. Rectal swabs and saliva samples were collected from symptomatic and asymptomatic students, and samples were also collected from the environment. Using PCR, followed by DNA sequencing, a single G9P[8] rotavirus strain was identified as the causative agent. The attack rate of the outbreak was 13.5% for boarders, which was significantly higher than that of day students (1.8%). Person-to-person transmission was the most plausible transmission mode. The HBGA phenotypes of the individuals in the study were determined by enzyme immunoassay, using saliva samples, while recombinant VP8* protein of the causative rotavirus strain was produced for HBGA binding assays to evaluate the host susceptibility. Our data showed that secretor individuals had a significantly higher risk of infection than nonsecretors. Accordingly, the VP8* protein bound nearly all secretor saliva samples, but not those of nonsecretors, explaining the observed infection of secretor individuals only. This is the first single-outbreak-based investigation showing that P[8] rotavirus infected only secretors. Our investigation also suggests that health education of school students is an important countermeasure against an outbreak of communicable disease.

Implementing the CRISPR/Cas9 Technology in Eucalyptus Hairy Roots Using Wood-Related Genes.

Eucalypts are the most planted hardwoods worldwide. The availability of the Eucalyptus grandis genome highlighted many genes awaiting functional characterization, lagging behind because of the lack of efficient genetic transformation protocols. In order to efficiently generate knock-out mutants to study the function of eucalypts genes, we implemented the powerful CRISPR/Cas9 gene editing technology with the hairy roots transformation system. As proofs-of-concept, we targeted two wood-related genes: Cinnamoyl-CoA Reductase1 (CCR1), a key lignin biosynthetic gene and IAA9A an auxin dependent transcription factor of Aux/IAA family. Almost all transgenic hairy roots were edited but the allele-editing rates and spectra varied greatly depending on the gene targeted. Most edition events generated truncated proteins, the prevalent edition types were small deletions but large deletions were also quite frequent. By using a combination of FT-IR spectroscopy and multivariate analysis (partial least square analysis (PLS-DA)), we showed that the CCR1-edited lines, which were clearly separated from the controls. The most discriminant wave-numbers were attributed to lignin. Histochemical analyses further confirmed the decreased lignification and the presence of collapsed vessels in CCR1-edited lines, which are characteristics of CCR1 deficiency. Although the efficiency of editing could be improved, the method described here is already a powerful tool to functionally characterize eucalypts genes for both basic research and industry purposes.

Induction of a high-titered antibody response using HIV gag-EV71 VP1-based virus-like particles with the capacity to protect newborn mice challenged with a lethal dose of enterovirus 71.

Enterovirus 71 (EV71) is the most frequently detected causative agent in hand, foot, and mouth disease (HFMD) and is a serious threat to public health in the Asia-Pacific region. Many EV71 vaccines are under development worldwide, and although both inactivated virus vaccines and virus-like particles (VLPs) are considered to be effective, the main focus has been on inactivated EV71vaccines. There have been very few studies on EV71 VLPs. In this study, using a strategy based on HIV gag VLPs, we constructed a gag-VP1 fusion gene to generate a recombinant baculovirus expressing the EV71 structural protein VP1 together with gag, which was then used to infect TN5 cells to form VLPs. The VLPs were characterized using transmission electron microscopy, electrophoresis and staining with Coomassie blue, and Western blotting. Mice immunized with gag-VP1 VLPs showed strong humoral and cellular immune responses. Finally, immunization of female mice with gag-VP1 VLPs provided effective protection of their newborn offspring against challenge with a lethal dose EV71. These results demonstrate a successful approach for producing EV71 VP1 VLPs based on the ability of HIV gag to self-assemble, thus providing a good foundation for producing high-titered anti-EV71 antibody by immunization with VLP-based gag EV71 VP1 protein.

Potential application of nanoliposomes loaded with complex tannins from the seed shell of Euryale ferox in the anti-browning of fresh-cut asparagus lettuce.

Surface browning of plant-derived fresh-cut products is mainly caused by conversion of the phenolic compounds into o-quinones under tyrosinase catalysis. In this study, the rarely reported complex tannins from Euryale ferox seed shell (ECTs) constituted by the units of 35.60% condensed tannins and 64.40% hydrolysable tannins were shown to suppress the activity of tyrosinase efficiently, supporting the exploitation of ECTs into novel anti-browning agents. However, the utilization of ECTs in food preservation is often restricted because of their chemical instability to external environment. Further fabrication of nanoliposomes loaded with ECTs (ECTs-NLs) herein was carried out to improve the stability of ECTs. DLS, TEM, FTIR, DSC and XRD confirmed that ECTs were encapsulated into nanoliposomes successfully, and ECTs-NLs appeared as vesicle-like spherical morphology with favorable encapsulation efficiency, uniform particle size distribution and negative zeta-potential. The resulting ECTs-NLs were relatively stable in the dark at 4 °C. Nanoliposomal encapsulation significantly enhanced ECTs stability, thus protecting inhibitory effect of ECTs against tyrosinase. Furthermore, anti-browning evaluation proved that ECTs-NLs had distinct advantages over free ECTs in alleviating surface browning of fresh-cut asparagus lettuces. These results suggested that nanoliposomes were effective in stabilizing ECTs and ECTs-NLs could be potentially applied to the fresh-cut food industry.

DUXAP8 Promotes LPS-Induced Cell Injury in Pulpitis by Regulating miR-18b-5p/HIF3A.

BACKGROUND: The dysregulated long noncoding RNAs (lncRNAs) are implicated in progression of various diseases, including pulpitis. Double homeobox A pseudogene 8 (DUXAP8) has been found to be upregulated in pulpitis. Herein, the functional mechanism of DUXAP8 in lipopolysaccharide (LPS)-induced pulpitis was explored. MATERIAL AND METHODS: DUXAP8, microRNA-18b-5p (miR-18b-5p), or hypoxia-inducible factor 3A (HIF3A) levels were examined through reverse transcription-quantitative polymerase chain reaction assay. Cell behaviours were determined by Cell Counting Kit-8 assay for cell viability, Ethynyl-2'-deoxyuridine (EdU) assay for cell proliferation, and flow cytometry for cell apoptosis. Protein levels were measured using western blot. Inflammatory reaction was analysed via enzyme-linked immunosorbent assay. Oxidative stress was assessed by commercial kits. Dual-luciferase reporter assay, RNA immunoprecipitation assay, and pull-down assay were used for validation of interaction between targets. RESULTS: Cell apoptosis, inflammatory reaction, and oxidative stress were induced by LPS in human dental pulp cells (HDPCs). DUXAP8 upregulation and miR-18b-5p downregulation were found in pulpitis. LPS-induced cell injury was relieved after downregulation of DUXAP8. DUXAP8 interacted with miR-18b-5p. The regulation of DUXAP8 was related to miR-18b-5p sponging function in LPS-treated HDPCs. HIF3A served as a target of miR-18b-5p. MiR-18b-5p protected against LPS-induced cell injury through targeting HIF3A. DUXAP8 targeted miR-18b-5p to regulate HIF3A level. CONCLUSIONS: Results demonstrated that LPS-induced cell injury in pulpitis was promoted by DUXAP8 through mediating miR-18b-5p/HIF3A axis.

Surfactant-assisted adsorption of uranyl ions in aqueous solution on TiO2/polythiophene nanocomposite.

In this work, hexadecyltrimethylammonium-bromide (HTAB)-modified polythiophene (PTh)/TiO2 nanocomposite (HTAB/PTh/TiO2) was applied to remove uranyl ions (UO22+). FT-IR, XRD, ζ potential, TGA, SEM, and XPS were utilized to obtain the chemical and physical properties of HTAB/PTh/TiO2. The effects of HTAB content, preparation temperature, and adsorption conditions on UO22+ removal were investigated comprehensively. And the UO22+ adsorption process on HTAB/PTh/TiO2 was fitted to the Sips model with a saturated adsorption capacity of 234.74 mg/g, which was 6 times over TiO2. The results suggested that the surfactant of HTAB can significantly improve the adsorption ability of TiO2 for UO22+ ions. This work provides a strategy of surfactant modification for enhancing the separation and recovery ability of adsorbent toward UO22+ in the radioactive wastewater.

Association between Dental Scaling and Reduced Risk of End-Stage Renal Disease: A Nationwide Matched Cohort Study.

Periodontitis is prevalent in patients with chronic kidney disease (CKD) and is also associated with kidney function decline. It is unclear whether dental scaling treatment prevents the progression of CKD. In a nationwide cohort study, Taiwan's National Health Insurance Research Database was used to select people with CKD. Propensity score-matching procedures were performed to compare the long-term risk of end-stage renal disease (ESRD) between CKD patients with and without the receipt of dental scaling. A total of 33,637 matched pairs with CKD were included, with 503,373 person-years of follow-up for analyses. Dental scaling was significantly associated with a lower risk of ESRD (adjusted hazard ratio (aHR): 0.83, 95% confidence interval (CI): 0.77-0.90). In addition, there was a dose-dependent relationship between the frequency of dental scaling and a reduced risk of ESRD. Dental scaling was also linked to reduced risks of major adverse cardiovascular events (aHR: 0.91, 95% CI: 0.87-0.95), sepsis (aHR: 0.81, 95% CI: 0.77-0.85), and all-cause mortality (aHR: 0.81, 95% CI: 0.76-0.87). Dental scaling was significantly associated with lower risks of progression to ESRD in patients with CKD. Regular dental scaling may serve as a prophylactic measure for kidney function decline.

An outbreak caused by GII.17 norovirus with a wide spectrum of HBGA-associated susceptibility.

During the past norovirus (NoV) epidemic season, a new GII.17 variant emerged as a predominant NoV strain, surpassed the GII.4 NoVs, causing outbreaks of acute gastroenteritis (AGE) in China. Here we report a study of an AGE outbreak in an elementary school in December 2014 caused by the new GII.17 NoV to explore the potential mechanism behind the sudden epidemics of the GII.17 NoV. A total of 276 individuals were sick with typical NoV infection symptoms of vomiting (93.4%), abdominal pain (90.4%), nausea (60.0%), and diarrhea (10.4%) at an attack rate of 5.7-16.9%. Genotyping of the symptomatic patients showed that individuals with a secretor positive status, including those with A, B, and O secretors and Lewis positive blood types, were sensitive to the virus, while the non-secretors and the Lewis negative individual were not. Accordingly, the recombinant capsid P protein of the GII.17 isolate showed a wide binding spectrum to saliva samples of all A, B, and O secretors. Thus, the broad binding spectrum of the new GII.17 variant could explain its widely spread nature in China and surrounding areas in the past two years.

Evidences for intertypic and intratypic recombinant events in EV71 of hand, foot and mouth disease during an epidemic in Hubei Province, China, 2011.

Humane enterovirus 71 (HEV 71) is a common contagious agent of hand, foot and mouth disease (HFMD) which is normally mild but can caused deaths and severe neurological complications. In April 2011, an unpredicted HFMD outbreak in Xiangyang City of Hubei Province in China resulted in a high aggregation of HFMD cases including fatal cases and many severe cases. In this study, 71 clinical specimens were collected according to the different symptoms and RNA extraction and RT-PCR amplification were performed immediately. Laboratory testing and genetic analyses were used to identify the casual pathogen of this outbreak. HEV71 was confirmed as the etiological pathogen of the outbreak. Similarity and phylogenetic analyses of the VP1 gene of HEV71 from Xiangyang showed that they belong to C4a cluster of the C4 subgenotype. Intertypic recombinant events were found in the 3D region between the Xiangyang HEV71 strains and Coxsackievirus A16 (CV-A16). Intratypic recombination was found in the 3D region between two same subgenotypic Xiangyang HEV71 strains in this outbreak. It is suggested that these recombination events played important roles in the emergence of the various HEV 71 subgenotypes and different type of recombination of HEV71 might exist in one outbreak which might be the reason for the different virulent HEV71 strains in an outbreak.

Treatment of alveolar cleft with distraction osteogenesis using anchorage with a tooth-microimplant joint in a dog model.

Our aim was to investigate the efficacy of correction of an alveolar cleft with distraction osteogenesis using anchorage with a tooth-microimplant joint in a canine model, which was established in 12 adult mongrel dogs that were subsequently randomised into two groups (n=6 in each). The first group comprised dogs that had osteogenesis using anchorage with a tooth (tooth group), while in the second, anchorage with tooth-microimplant joint (microimplant group) was used. All animals were killed one month after completion of distraction. Samples were collected for gross observation and histological examination. There was a significant difference in the degree of movement of the anchorage teeth in the transport discs between the 2 groups (p<0.01). There was less prominent inclination and shift of the natural teeth in the transport disc and less bony resorption around the root in the microimplant group than in the tooth group. These changes were less remarkable in the microimplant group. Treatment of alveolar cleft by distraction osteogenesis using anchorage with a tooth-microimplant joint is practical, and yields better results.

[Relationship between masseter muscle electromyographic activities and the position of the mandible].

OBJECTIVE: To investigate the changes in electromyographic activities of the temporal and masseter muscles at different positions of the mandible. METHODS: Twenty orthodontic patients with Angle Class II malocclusion and mandibular retrusion (ANB<6°) aged 10-14 years were enrolled in this study. All the patients were treated with Forsus fixed functional appliance combined with MBT straight-wire appliance. The electromyographic activities of the temporal (T) and masseter (M) muscles before, during and after functional treatment were evaluated by assessing the average integrated electromyogram (EMG) and T/M ratio at clenching status in different mandibular positions. RESULTS: After functional forward positioning of the mandible, the electromyographic activities of the temporal and masseter muscles decreased and T/M ratio increased significance at the clenching status (P<0.05). CONCLUSION: The appliance insertion and activation is associated with a decreased EMG activity of the temporal and masseter muscles, and the T/M ratio is correlated to the position of the mandible.

[Maxillary sinus augmentation with simultaneous implantation using guided bone regeneration].

OBJECTIVE: To investigate the effect of new bone formation in sinus augmentation with guided bone regeneration (GBR) using collagen membranes. METHODS: The first maxillary molars of 18 adult female Beagle dogs were extracted and the sinus floors of both sides were lifted with simultaneous implantation. A combination of autografts and Bio-Oss in a 2:1 ratio was placed in the space under the membrane. On the experimental side in each dog, the collagen membrane was folded at the lateral osteotomy window, the apex of the implants and a certain part of palatal bone. On the contralateral control side, the collagen membrane only covered the osteotomy window. Six animals were sacrificed at 4, 12, and 24 weeks respectively after surgery. Gross observation, biomechanical testing and histological examinations were performed. RESULTS: The translocation of grafted materials and bone absorption were found on the top of implants in the control side, and the grafted materials kept original shape at the experimental side at 4th week. The granule of Bio-oss absorbed obviously at 12th and 24th week. The pull-out force increased with time. At 24th week, the force of pull out was 558.1 ± 37.4 N at the study side, and 471.4 ± 31.5 N at the control side. There was a significant difference in the pull-out force was noted between the two groups (P < 0.01). Histological examination showed new bone formation on the sinus floor, and the grafted materials gradually reduced with time CONCLUSIONS: GBR with the enfolded-coverage of the membrane can effectively decrease absorption of the grafted materialon the apical surface of implants and stimulate new bone formation in the sinus augmentation.