My 20 years together with Journal of Oral Pathology and Medicine.

Shan Gao has served at Journal of Oral Pathology and Medicine (JOPM) for 20 years, and currently as Associate Editor of JOMP, Beijing, China. After he finished his 8 year education in Stomatology for both bachelor and master degrees, he started 10 years of clinic practice in Endodontics and Oral Medicine in China, followed by 12 years basic research work in Molecular Biology, including 3 years for a PhD degree in Denmark and 10 years industry experience in a leading RNAi therapeutic company in China. During those years of experience, he built up a close relationship with Oral Pathology and Oral Medicine. It is great opportunity to introduce his story together with JOPM, accompanying with his personal research experience, at the moment of the 50th Anniversary of JOPM.

Temporal stability and assembly mechanisms of gut microbiota in sea cucumbers response to nanoplastics treatment.

Aquaculture provides essential food for humans, and the health of farmed species is particularly important for the aquaculture industry. Aquaculture environment could be a sink of plastic debris (PDs) due to the enclosed character and heavy use of plastics. Gut microbiota of aquaculture species could respond to the exogenous pollutants and regulate the health of hosts. Here, variations in gut microbiota of Apostichopus japonicus induced by the ingested nanoplastics (NPs) were investigated by a lab experiment. We selected a NPs concentration gradient of 100 mg/kg and 500 mg/kg to simulate microplastic pollution to A. japonicus, and the significant differences in gut microbiota composition after 21 days of NP exposure were evaluated. According to the high-throughput sequencing from time series samples, a decrease of diversity in gut microbiota of A. japonicus with dietary NPs was observed. In addition, the gut microbiota compositions of sea cucumbers with and without NPs exposure were also distinct, expressing as enrichment of Bacteroidota while reducement of Proteobacteria under NPs stresses. Combined the results of network analysis, the less complexity and stability of gut microbiota in sea cucumbers with dietary NPs were proved. Based on the neutral community model, the ingested NPs elevated the contribution of stochastic processes for the gut microbiota assembly in sea cucumbers. Our study showed that substantial variations in gut microbiota of A. japonicus under NPs stresses, and also explored the underlying mechanisms regulating these changes. This research would offer new meaningful insights into the toxicity of NPs on sea cucumbers, contributing a solid fundament to improve the health of sea cucumbers under NPs stresses.

Bone Tumor Suppression in Rabbits by Hyperthermia below the Clinical Safety Limit Using Aligned Magnetic Bone Cement.

Demonstrating highly efficient alternating current (AC) magnetic field heating of nanoparticles in physiological environments under clinically safe field parameters has remained a great challenge, hindering clinical applications of magnetic hyperthermia. In this work, exceptionally high loss power of magnetic bone cement under the clinical safety limit of AC field parameters, incorporating direct current field-aligned soft magnetic Zn0.3 Fe2.7 O4 nanoparticles with low concentration, is reported. Under an AC field of 4 kA m-1 at 430 kHz, the aligned bone cement with 0.2 wt% nanoparticles achieves a temperature increase of 30 °C in 180 s. This amounts to a specific loss power value of 327 W gmetal-1 and an intrinsic loss power of 47 nHm2 kg-1 , which is enhanced by 50-fold compared to randomly oriented samples. The high-performance magnetic bone cement allows for the demonstration of effective hyperthermia suppression of tumor growth in the bone marrow cavity of New Zealand White Rabbits subjected to rapid cooling due to blood circulation, and significant enhancement of survival rate.

Long-term effectiveness of lycopene in the management of oral submucous fibrosis (OSMF): A 3-years follow-up study.

BACKGROUND: Long-term follow-ups after receiving lycopene therapy for management of oral submucous fibrosis (OSMF) are scarce. The study aimed to assess the long-term efficacy of lycopene for management of OSMF symptoms. METHODS: In this prospective clinical study, 400 clinically diagnosed early OSMF patients were assessed for the efficacy of lycopene in alleviation of burning sensation (BS) and reduced mouth opening (MO) symptoms in comparison to placebo. After 1-year follow-up, group A (lycopene group) was divided equally into group A1 and group A2. group A1 patients were retreated with lycopene and the A2 group was followed without retreatment. After 2-year follow-up, the Group A2 patients were advised retreatment but not followed as most of the patient did not agree for follow-up. However, group A1 patients were continued to follow-up every 6 months for a total three years. Statistical analysis was by independent sample t-test and P-value <.05 were considered as significant. RESULTS: A statistically significant difference (P < .05) in BS and MO was found between group A and B with lycopene showing better results. At one-year follow-up, a statistically significant recurrence in the symptoms was found (P < .05) in the lycopene group (group A). After the second intervention, there was a statistically significant difference in the improvement of symptoms between the group A1 and A2 at 6 months and 1 year (P < .05) with group A1 (retreatment) showing better results. CONCLUSIONS: Treatment with lycopene led to improvement in the symptoms of OSMF in the present study. The results highlight the importance of retreatment of lycopene for its long-term effect on alleviation the symptoms of OSMF.

Ladder chain Cd-based polymer as a highly effective adsorbent for removal of Congo red.

Developing of high effective and fast-rate adsorbent materials has been recently attracted intensive attentions all over the world due to organic dye polluted water treatment. However, few studies have been reported on the ultrahigh-capacity and fast-rate removal of Congo red. In this work, a new stable Cd-based coordination polymer exhibits excellent adsorption performance towards Congo Red. This ladder chain [Cd4(H2L)4(H2O)8(NDS)]n·3n(NDS) (I) (H2L = N1,N2-bis(pyridin-3-ylmethyl) ethane-1,2-diamine, 1,5-H2NDS = 1,5-naphthalene disulfonic acid) has been successfully synthesized by the hydrothermal reaction. At room temperature, the experimental adsorption capacity of coordination polymer (I) towards Congo red can reach up to 16,880 mg g-1 in 20 min (pH = 2.0-3.2), and its higher capacity and faster rate are all better than those in reported inorganic and metal-organic frameworks absorbents. The adsorption process is spontaneous and endothermic reaction, and fits well with the second-order kinetics, Langmuir and Scatchard isotherm adsorption models. The excellent adsorption performance of (I) towards Congo red is related to the strong electrostatic, various hydrogen bonding and π-π stacking interactions under acidic conditions.

Near-infrared light-responsive nanoparticles for improved anticancer efficacy through synergistic chemo-photothermal therapy.

Combined treatment is more effective than single treatment against most forms of cancer. The synergistic chemo-thermotherapy mediated by nanoparticles has superior advantages of lesser adverse effects as a promising cancer therapy modality. In this study, we report a theranostic carrier system co-encapsulating Doxorubicin (DOX) and Indocyanine green (ICG) into the D-α-Tocopheryl polyethylene glycol 1000 succinate (TPGS). Full physicochemical characterization studies of the DOX/ICG-loaded TPGS nanoparticles (TNPs) are performed. TNPs have a mean size around 60 nm with superior photostability, and entrapment efficiency of drugs in TNPs was 75.0% for ICG and 68.3% for DOX. TNPs also exhibit a longer sustained release with around 63% of the entrapped drug in 24 h. In vitro studies, TNPs could effectively enhance cellular uptake of DOX and ICG, which permitted high therapeutic efficacy against cancer cells. Further, we investigate antitumor efficacy of TNPs along with its impact on major organs in vivo, TNPs also exhibit a complete inhibition of tumor growth and minimal side effects after irradiation. Collectively, these results suggest that near-infrared light-responsive TNPs can further enhance antitumor effects by synergistic chemo-photothermal therapy.

TBX1 protein interactions and microRNA-96-5p regulation controls cell proliferation during craniofacial and dental development: implications for 22q11.2 deletion syndrome.

T-box transcription factor TBX1 is the major candidate gene for 22q11.2 deletion syndrome (22q11.2DS, DiGeorge syndrome/Velo-cardio-facial syndrome), whose phenotypes include craniofacial malformations such as dental defects and cleft palate. In this study, Tbx1 was conditionally deleted or over-expressed in the oral and dental epithelium to establish its role in odontogenesis and craniofacial developmental. Tbx1 lineage tracing experiments demonstrated a specific region of Tbx1-positive cells in the labial cervical loop (LaCL, stem cell niche). We found that Tbx1 conditional knockout (Tbx1(cKO)) mice featured microdontia, which coincides with decreased stem cell proliferation in the LaCL of Tbx1(cKO) mice. In contrast, Tbx1 over-expression increased dental epithelial progenitor cells in the LaCL. Furthermore, microRNA-96 (miR-96) repressed Tbx1 expression and Tbx1 repressed miR-96 expression, suggesting that miR-96 and Tbx1 work in a regulatory loop to maintain the correct levels of Tbx1. Cleft palate was observed in both conditional knockout and over-expression mice, consistent with the craniofacial/tooth defects associated with TBX1 deletion and the gene duplication that leads to 22q11.2DS. The biochemical analyses of TBX1 human mutations demonstrate functional differences in their transcriptional regulation of miR-96 and co-regulation of PITX2 activity. TBX1 interacts with PITX2 to negatively regulate PITX2 transcriptional activity and the TBX1 N-terminus is required for its repressive activity. Overall, our results indicate that Tbx1 regulates the proliferation of dental progenitor cells and craniofacial development through miR-96-5p and PITX2. Together, these data suggest a new molecular mechanism controlling pathogenesis of dental anomalies in human 22q11.2DS.

Oral submucous fibrosis: An update on current theories of pathogenesis.

Over the last 40 years, many theories linking oral submucous fibrosis (OSMF) to various risk factors have been proposed. Spicy, pungent foods and irritants such as supari (areca nut), paan (betel leaves), tobacco (through chewing or smoking)-the common Asian habits of chewing the aforementioned agents-have all been incriminated as causative agents. Systemic factors such as nutritional deficiency, genetic predisposition and autoimmunity have also been proposed in the pathogenesis of OSMF. However, the precise aetiology of OSMF is still unknown, and no conclusive evidence has been found despite many extensive investigations on implicated factors. Most of the ideas proposed have been derived from the existing clinical and epidemiological data. We present a comprehensive review of the various theories regarding the pathogenesis of the condition, but have not concentrated on malignant transformation in this article.

LncRNA H19 and Target Gene-mediated Cleft Palate Induced by TCDD.

This study investigated the role of long non-coding RNAs (lncRNAs) in the development of the palatal tissues. Cleft palates in mice were induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Expression levels of long non-coding RNA H19 (lncRNA H19) and insulin-like growth factor 2 (IGF2) gene were measured by quantitative real-time polymerase chain reaction (qRT-PCR). The rate of occurrence of cleft palate was found to be 100% by TCDD exposure, and TCDD could cause short upper limb, cerebral fissure, webbed neck, and short neck. The expression levels of lncRNA H19 and IGF2 gene specifically showed embryo age-related differences on E13, E14, and E15 in the palatal tissues. The expression levels of lncRNA H19 and IGF2 gene showed an inverse relationship on E13, E14, and E15. These findings demonstrated that lncRNA H19 and IGF2 can mediate the development of mouse cleft palate.

A pituitary homeobox 2 (Pitx2):microRNA-200a-3p:ß-catenin pathway converts mesenchymal cells to amelogenin-expressing dental epithelial cells.

Pitx2, Wnt/ß-catenin signaling, and microRNAs (miRs) play a critical role in the regulation of dental stem cells during embryonic development. In this report, we have identified a Pitx2:ß-catenin regulatory pathway involved in epithelial cell differentiation and conversion of mesenchymal cells to amelogenin expressing epithelial cells via miR-200a. Pitx2 and ß-catenin are expressed in the labial incisor cervical loop or epithelial stem cell niche, with decreased expression in the differentiating ameloblast cells of the mouse lower incisor. Bioinformatics analyses reveal that miR-200a-3p expression is activated in the pre-ameloblast cells to enhance epithelial cell differentiation. We demonstrate that Pitx2 activates miR-200a-3p expression and miR-200a-3p reciprocally represses Pitx2 and ß-catenin expression. Pitx2 and ß-catenin interact to synergistically activate gene expression during odontogenesis and miR-200a-3p attenuates their expression and directs differentiation. To understand how this mechanism controls cell differentiation and cell fate, oral epithelial and odontoblast mesenchymal cells were reprogrammed by a two-step induction method using Pitx2 and miR-200a-3p. Conversion to amelogenin expressing dental epithelial cells involved an up-regulation of the stem cell marker Sox2 and proliferation genes and decreased expression of mesenchymal markers. E-cadherin expression was increased as well as ameloblast specific factors. The combination of Pitx2, a regulator of dental stem cells and miR-200a converts mesenchymal cells to a fully differentiated dental epithelial cell type. This pathway and reprogramming can be used to reprogram mesenchymal or oral epithelial cells to dental epithelial (ameloblast) cells, which can be used in tissue repair and regeneration studies.

Underwater drag-reducing effect of superhydrophobic submarine model.

To address the debates on whether superhydrophobic coatings can reduce fluid drag for underwater motions, we have achieved an underwater drag-reducing effect of large superhydrophobic submarine models with a feature size of 3.5 cm × 3.7 cm × 33.0 cm through sailing experiments of submarine models, modified with and without superhydrophobic surface under similar power supply and experimental conditions. The drag reduction rate reached as high as 15%. The fabrication of superhydrophobic coatings on a large area of submarine model surfaces was realized by immobilizing hydrophobic copper particles onto a precross-linked polydimethylsiloxane (PDMS) surface. The pre-cross-linking time was optimized at 20 min to obtain good superhydrophobicity for the underwater drag reduction effect by investigating the effect of pre-cross-linking on surface wettability and water adhesive property. We do believe that superhydrophobic coatings may provide a promising application in the field of drag-reducing of vehicle motions on or under the water surface.

Macrophage-mediated nanoparticle delivery to the periodontal lesions in established murine model via Pg-LPS induction.

We established a murine periodontitis model by local injection of lipopolysaccharide of Porphyromonas gingivalis (Pg-LPS) into the gingival sulcus of mandibular left incisor four times with 48-h interval. The histological examination of the periodontal tissues demonstrated that significant loss of periodontal bone and ligaments was observed in the lesion side with abundant inflammatory cell infiltration. Two days after the last injection, Cy5-labelled siRNA/chitosan particles were injected intraperitoneally (ip). The chitosan/siRNA particles were taken up by peritoneal macrophages, which subsequently migrated to the inflamed gingival area evaluated by in vivo imaging. The localization of macrophages in the inflamed region was further confirmed by immunofluorescent staining. The present report demonstrates that intragingival injection of Pg-LPS can be used to create an experimental model of periodontal inflammation in mice and that recruitment of macrophages with chitosan/siRNA nanoparticles to the inflamed area opens the possibility of an RNAi-based therapeutic approach using chitosan as a carrier in periodontitis.

Focal adhesion kinase knockdown in carcinoma-associated fibroblasts inhibits oral squamous cell carcinoma metastasis via downregulating MCP-1/CCL2 expression.

Carcinoma-associated fibroblasts (CAFs) have been demonstrated to play an important role in the occurrence and development of oral squamous cell carcinoma (OSCC). The aim of this study is to investigate the influence of CAFs on OSCC cells and to explore the role of focal adhesion kinase (FAK) in this process. The results showed that oral CAFs expressed a higher level of FAK than normal human gingival fibroblasts (HGFs), and the conditioned medium (CM) of CAFs could induce the invasion and migration of SCC-25, one oral squamous carcinoma cell line. However, knockdown of FAK by small interfering RNA (siRNA) resulted in inhibition of CAF-CM induced cell invasion and migration in SCC-25, probably by reducing the production of monocyte chemoattractant protein-1 (MCP-1/CCL2), one of downstream target chemokines. Therefore, our findings indicated that targeting FAK in CAFs might be a promising strategy for the treatment of OSCC in the future.

Protein inhibitors of activated STAT (Pias1 and Piasy) differentially regulate pituitary homeobox 2 (PITX2) transcriptional activity.

Protein inhibitors of activated STAT (Pias) proteins can act independent of sumoylation to modulate the activity of transcription factors and Pias proteins interacting with transcription factors can either activate or repress their activity. Pias proteins are expressed in many tissues and cells during development and we asked if Pias proteins regulated the pituitary homeobox 2 (PITX2) homeodomain protein, which modulates developmental gene expression. Piasy and Pias1 proteins are expressed during craniofacial/tooth development and directly interact and differentially regulate PITX2 transcriptional activity. Piasy and Pias1 are co-expressed in craniofacial tissues with PITX2. Yeast two-hybrid, co-immunoprecipitation and pulldown experiments demonstrate Piasy and Pias1 interactions with the PITX2 protein. Piasy interacts with the PITX2 C-terminal tail to attenuate its transcriptional activity. In contrast, Pias1 interacts with the PITX2 C-terminal tail to increase PITX2 transcriptional activity. The E3 ligase activity associated with the RING domain in Piasy is not required for the attenuation of PITX2 activity, however, the RING domain of Pias1 is required for enhanced PITX2 transcriptional activity. Bimolecular fluorescence complementation assays reveal PITX2 interactions with Piasy and Pias1 in the nucleus. Piasy represses the synergistic activation of PITX2 with interacting co-factors and Piasy represses Pias1 activation of PITX2 transcriptional activity. In contrast, Pias1 did not affect the synergistic interaction of PITX2 with transcriptional co-factors. Last, we demonstrate that Pias proteins form a complex with PITX2 and Lef-1, and PITX2 and ß-catenin. Lef-1, ß-catenin, and Pias interactions with PITX2 provide new molecular mechanisms for the regulation of PITX2 transcriptional activity and the activity of Pias proteins.

Chitosan hydrogel as siRNA vector for prolonged gene silencing.

BACKGROUND: The periodontitis is one of the most prevalent diseases with alveolar resorption in adult people and is the main cause of the tooth loss. To investigate the possibility for protecting the loss of alveolar bone in periodontal diseases, a RNAi-based therapeutic strategy is applied for silencing RANK signaling using thermosensitive chitosan hydrogel as siRNA reservoir and vector. RESULTS: The thermosensitive chitosan hydrogel was formed from solution (PH = 7.2, at 4°C) at 37°C within 8 minutes. The degradation rates of hydrogel were ~50% and 5% (W remaining/W beginning) in the presence and absence of lysozyme, respectively, over a period of 20 days. The concurrent cumulative in vitro release of Cy3-labeled siRNA from the hydrogel was 50% and 17% over 14 days, with or without lysozyme digestion, respectively. High cell viability (>88%) was maintained for cells treated with hydrogel loaded with RANK specific siRNA and RANK knockdown was prolonged for up to 9 days when cells were incubated with siRNA/hydrogel complex. In vivo release of siRNA was investigated in a subcutaneous delivery setup in mice. The fluorescent signal from siRNA within hydrogel was remained for up to 14 days compared to less than one day for siRNA alone. CONCLUSIONS: Chitosan hydrogel can potentially serve as a suitable reservoir and vector for local sustained delivery of siRNA in potential therapy.

Simultaneous detection of enrofloxacin and florfenicol in animal-derived foods based on fluorescence quenching BELISA and a nanozyme catalytic strategy.

Enrofloxacin (ENRO) and florfenicol (FF) are animal-specific drugs, but they present great harm to human health. Therefore, it is essential to rapidly and accurately detect ENRO and FF in animal-derived foods simultaneously. Herein, dual-template molecular imprinted polymers (MIPs) with specific recognition of ENRO and FF were prepared, meanwhile, the molar ratios of templates to monomer and cross-linker were optimized and then applied as a bionic antibody to experiment. Based on the principle that the fluorescence of QDs could be efficiently quenched by the enzymatic fabrication of Prussian blue nanoparticles (PBNPs), a novel and sensitive fluorescence quenching biomimetic enzyme-linked immunosorbent assay (BELISA) was established for simultaneous detection of ENRO and FF by the conversion of the absorption signal into fluorescent signals. Under optimal conditions, the detection limit (IC15) was 4.64 ng L-1 for ENRO and 1.33 ng L-1 for FF. Besides, matrix interference of chicken, eggs, milk and shrimp samples, was investigated in our study, and the result indicates that all of the sample matrices had a profound impact on the fluorescence of QDs, especially for milk samples (with Im of 94.10 %). After performing the matrix-elimination experiments, chicken, eggs, milk and shrimp samples spiked with ENRO and FF were extracted and detected by this proposed method, with recoveries ranging from 82.70 to 113.48 %. The results correlated well with those obtained using HPLC. In conclusion, the developed method could be an alternative and sensitive method for the simultaneous detection of ENRO and FF in animal-derived foods.

A biodegradable pH-response packaging film with blueberry extract: Blown-extrusion fabrication, multifunctional activity, and kinetic investigation.

This work proposed a novel strategy for manufacturing biodegradable pH-response packaging. Briefly, to minimize the amount and thermal processing times of blueberry extract (BE), ethanol-dissolved BE (≤ 3‰ w/w) was sprayed onto the starch/poly(butylene adipate-co-terephthalate) (PBAT) pellets before extrusion blowing. BE was well-integrated into the matrix, forming uniformly colored films. The films with BE exhibited superior mechanical (7.85 MPa of strength, 606.53% of elongation) and enhanced barrier capabilities against ultraviolet light, moisture, and gas. Additionally, they exhibited good antioxidant capacity (68.69%), antibacterial activity (72.40%), and maintained color stability. The film with 3‰ w/w BE presented excellent color responsiveness (ΔE⁎ ≥ 15) in the alkaline range, and successfully monitored the spoilage of shrimp. The pigments in the film had the maximum migration degree (≥ 70%) and rate in 50% ethanol simulation, following a first-order kinetic behavior dominated by Fickian diffusion. Findings supported the application of this strategy in the fabrication of starch/PBAT/BE films for pH-response intelligent packaging.

Starch as a smart, cheap, and green gatekeeper for the controlled release of propyl gallate from antioxidant biodegradable packaging films.

Oxidative rancidity of food products and massive consumption of plastic packaging have put the necessity in manufacturing novel antioxidant biodegradable packaging films. A comprehensive investigation was conducted on starch/poly(butylene adipate-co-terephthalate) (PBAT) antioxidant blown films, in which starch acted as a gatekeeper for the controlled release of propyl gallate (PG). PG was well integrated into the matrices and bound to starch molecules by hydrogen bonding. All films showed strong anti-ultraviolet performance, and higher oxygen barrier than the traditional polyethylene film. Increasing starch proportions promoted the swelling of films and the release of PG, thereby causing higher antioxidant activity at the same contact time to free radical solutions. Similar polarity made PG prone to partition and rapid migration into the food simulants with higher ethanol concentration and the high-fat-content peanut butter. The film with 20:80 w/w starch/PBAT proportion and 3% w/w PG content effectively suppressed the oxidation of peanut butter within 300-day storage. Findings demonstrated this strategy for manufacturing starch/PBAT antioxidant films as a long-term active packaging in food industry.

Coxsackievirus A10 impairs nail regeneration and induces onychomadesis by mimicking DKK1 to attenuate Wnt signaling.

Coxsackievirus A10 (CV-A10) infection, a prominent cause of childhood hand-foot-and-mouth disease (HFMD), frequently manifests with the intriguing phenomenon of onychomadesis, characterized by nail shedding. However, the underlying mechanism is elusive. Here, we found that CV-A10 infection in mice could suppress Wnt/ß-catenin signaling by restraining LDL receptor-related protein 6 (LRP6) phosphorylation and ß-catenin accumulation and lead to onychomadesis. Mechanistically, CV-A10 mimics Dickkopf-related protein 1 (DKK1) to interact with Kringle-containing transmembrane protein 1 (KRM1), the CV-A10 cellular receptor. We further found that Wnt agonist (GSK3ß inhibitor) CHIR99021 can restore nail stem cell differentiation and protect against nail shedding. These findings provide novel insights into the pathogenesis of CV-A10 and related viruses in onychomadesis and guide prognosis assessment and clinical treatment of the disease.