Cross-kingdom RNA interference mediated by insect salivary microRNAs may suppress plant immunity.

Communication between insects and plants relies on the exchange of bioactive molecules that traverse the species interface. Although proteinic effectors have been extensively studied, our knowledge of other molecules involved in this process remains limited. In this study, we investigate the role of salivary microRNAs (miRNAs) from the rice planthopper Nilaparvata lugens in suppressing plant immunity. A total of three miRNAs were confirmed to be secreted into host plants during insect feeding. Notably, the sequence-conserved miR-7-5P is specifically expressed in the salivary glands of N. lugens and is secreted into saliva, distinguishing it significantly from homologues found in other insects. Silencing miR-7-5P negatively affects N. lugens feeding on rice plants, but not on artificial diets. The impaired feeding performance of miR-7-5P-silenced insects can be rescued by transgenic plants overexpressing miR-7-5P. Through target prediction and experimental testing, we demonstrate that miR-7-5P targets multiple plant genes, including the immune-associated bZIP transcription factor 43 (OsbZIP43). Infestation of rice plants by miR-7-5P-silenced insects leads to the increased expression of OsbZIP43, while the presence of miR-7-5P counteracts this upregulation effect. Furthermore, overexpressing OsbZIP43 confers plant resistance against insects which can be subverted by miR-7-5P. Our findings suggest a mechanism by which herbivorous insects have evolved salivary miRNAs to suppress plant immunity, expanding our understanding of cross-kingdom RNA interference between interacting organisms.

Ectopic Activation of Fgf8 in Dental Mesenchyme Causes Incisor Agenesis and Molar Microdontia.

Putatively, tooth agenesis was attributed to the initiation failure of tooth germs, though little is known about the histological and molecular alterations. To address if constitutively active FGF signaling is associated with tooth agenesis, we activated Fgf8 in dental mesenchyme with Osr-cre knock-in allele in mice (Osr2-creKI; Rosa26R-Fgf8) and found incisor agenesis and molar microdontia. The cell survival assay showed tremendous apoptosis in both the Osr2-creKI; Rosa26R-Fgf8 incisor epithelium and mesenchyme, which initiated incisor regression from cap stage. In situ hybridization displayed vanished Shh transcription, and immunostaining exhibited reduced Runx2 expression and enlarged mesenchymal Lef1 domain in Osr2-creKI; Rosa26R-Fgf8 incisors, both of which were suggested to enhance apoptosis. In contrast, Osr2-creKI; Rosa26R-Fgf8 molar germs displayed mildly suppressed Shh transcription, and the increased expression of Ectodin, Runx2 and Lef1. Although mildly smaller than WT controls prenatally, the Osr2-creKI; Rosa26R-Fgf8 molar germs produced a miniature tooth with impaired mineralization after a 6-week sub-renal culture. Intriguingly, the implanted Osr2-creKI; Rosa26R-Fgf8 molar germs exhibited delayed odontoblast differentiation and accelerated ameloblast maturation. Collectively, the ectopically activated Fgf8 in dental mesenchyme caused incisor agenesis by triggering incisor regression and postnatal molar microdontia. Our findings reported tooth agenesis resulting from the regression from the early bell stage and implicated a correlation between tooth agenesis and microdontia.

Advances in the applications of mesenchymal stem cell-conditioned medium in ocular diseases.

Mesenchymal stem cell-conditioned medium (MSC-CM), also known as secretome, is secreted by MSC and contains a variety of bioactive factors with anti-inflammatory, anti-apoptotic, neuroprotection, and proliferation effects. Increasing evidence proved that MSC-CM plays an important role in various diseases, including skin, bone, muscle, and dental diseases. However, the role of MSC-CM in ocular diseases is not quite clear, Therefore, this article reviewed the composition, biological functions, preparation, and characterization of MSC-CM and summarized current research advances in different sources of MSC-CM in corneal and retinal diseases, including dry eye, corneal epithelial damage, chemical corneal injury, retinitis pigmentosa (RP), anterior ischemic optic neuropathy (AION), diabetic retinopathy (DR), and other retinal degenerative changes. For these diseases, MSC-CM can promote cell proliferation, reduce inflammation and vascular leakage, inhibit retinal cell degeneration and apoptosis, protect corneal and retinal structures, and further improves visual function. Hence, we summarize the production, composition and biological functions of MSC-CM and focus on describing its mechanisms in the treatment of ocular diseases. Furthermore, we look at the unexplored mechanisms and further research directions for MSC-CM based therapy in ocular diseases.

Deficiency of Fam20b-Catalyzed Glycosaminoglycan Chain Synthesis in Neural Crest Leads to Cleft Palate.

Cleft palate is one of the most common birth defects. Previous studies revealed that multiple factors, including impaired intracellular or intercellular signals, and incoordination of oral organs led to cleft palate, but were little concerned about the contribution of the extracellular matrix (ECM) during palatogenesis. Proteoglycans (PGs) are one of the important macromolecules in the ECM. They exert biological functions through one or more glycosaminoglycan (GAG) chains attached to core proteins. The family with sequence similarity 20 member b (Fam20b) are newly identified kinase-phosphorylating xylose residues that promote the correct assembly of the tetrasaccharide linkage region by creating a premise for GAG chain elongation. In this study, we explored the function of GAG chains in palate development through Wnt1-Cre; Fam20bf/f mice, which exhibited complete cleft palate, malformed tongue, and micrognathia. In contrast, Osr2-Cre; Fam20bf/f mice, in which Fam20b was deleted only in palatal mesenchyme, showed no abnormality, suggesting that failed palatal elevation in Wnt1-Cre; Fam20bf/f mice was secondary to micrognathia. In addition, the reduced GAG chains promoted the apoptosis of palatal cells, primarily resulting in reduced cell density and decreased palatal volume. The suppressed BMP signaling and reduced mineralization indicated an impaired osteogenesis of palatine, which could be rescued partially by constitutively active Bmpr1a. Together, our study highlighted the key role of GAG chains in palate morphogenesis.

[Effect of Lipoteichoic Acid Synthesis-Related Gene dltD on Acid Tolerance of Highly Cariogenic Strains of Streptococcus mutans].

Objective: To study the role and possible mechanism of dltD in the acid tolerance of Streptococcus mutans 593 (SM593), and to provide a theoretical basis for the ecological prevention and control of dental caries by constructing the dltD gene deletion strain of SM593 (SM593-ΔdltD). Methods: 1) SM593-Δ dltD was constructed by homologous recombination. 2) The growth curve of SM593 dltD and SM593-Δ dltD under different pH culture conditions was drawn by the automatic growth curve analyzer to compare their acid tolerance. Colony forming unit (CFU) at different time points was used to calculate the survival rate and to compare the acid tolerance response (ATR) of SM593 and SM593-Δ dltD. 3) Under different pH conditions, glycolysis experiments, proton permeability test and H +-ATPase activity test were conducted to make preliminary exploration into the mechanisms of how dltD gene deletion may affect acid tolerance. Results: 1) PCR and sequencing results showed that the SM593-Δ dltD was constructed successfully. 2) With decreasing pH value of the culture medium, the growth of SM593-Δ dltD slowed down. When the pH value of the culture medium was 5.0, SM593-Δ dltD was not allowed to grow, and its acid tolerance was lower than that of SM593. Compared with SM593, the ATR capability of SM593-Δ dltD was decreased. 3) SM593 dltD and SM593-Δ dltD did not show obvious difference in their glycolysis ability under different pH conditions. Compared with SM593 dltD, the proton permeability of SM593-Δ dltD under different pH conditions was increased significantly (P<0.05), and H +-ATPase activity decreased significantly (P<0.05). Conclusion: Compared with SM593 dltD, SM593-Δ dltD showed obvious decrease in acid tolerance, which may be caused by the significant increase in proton permeability and significant decrease in the H +-ATPase activity induced by the deletion of the dltD gene, hence reducing its ability to maintain intracellular pH homeostasis.

Tissue interactions are indispensable for cavity formation and disc separation in the temporomandibular joint.

Purpose: Our previous study found that in the temporomandibular joint (TMJ) of the K14-cre; Ctnnb1ex3f mouse embryo, the morphogenesis of glenoid fossa was interrupted by the dislocated condyle. This observation suggested that the formation of the glenoid fossa required tissue interactions with condylar mesenchyme. The purpose of this study was to clarify if the interactions between other components are essential for TMJ morphogenesis.Materials and methods: We examined the gross morphology, histology, cell proliferation, and gene expression in the developing TMJ of K14-cre; Ctnnb1ex3f mice by whole-mount bone and cartilage staining, Azon staining, BrdU labeling, and in situ hybridization, respectively.Results: In K14-cre; Ctnnb1ex3f mice, the zygomatic arch was misconnected to the mandibular bone by ectopic bone formation, which disrupted the attachment of temporalis to the mandibular bone and joint capsule formation. Although the initiation and differentiation of the condylar cartilage were slightly impacted, the K14-cre; Ctnnb1ex3f TMJ lacked joint cavities and separated disc, suggesting that the tissue interactions between the joint capsule and the TMJ were indispensable for the cavity formation and disc separation. The ectopic activation of Gli2 in the cells occupying the cavities, and the enhanced PTHrP transcription in the condylar perichondrium of the K14-cre; Ctnnb1ex3f TMJ suggested that the disrupted interactions between the joint capsule and the TMJ impaired cavity formation and disc separation by altering Hh signaling.Conclusion: Joint capsule formation was essential for cavity formation and disc separation during TMJ development.

Effect of Side Chain Substituent Volume on Thermoelectric Properties of IDT-Based Conjugated Polymers.

A p-type thermoelectric conjugated polymer based on indacenodithiophene and benzothiadiazole is designed and synthesized by replacing normal aliphatic side chains (P1) with conjugated aromatic benzene substituents (P2). The introduced bulky substituent on P2 is detrimental to form the intensified packing of polymers, therefore, it hinders the efficient transporting of the charge carriers, eventually resulting in a lower conductivity compared to that of the polymers bearing aliphatic side chains (P1). These results reveal that the modification of side chains on conjugated polymers is crucial to rationally designed thermoelectric polymers with high performance.

Fluorescence Sensor Based on Biosynthetic CdSe/CdS Quantum Dots and Liposome Carrier Signal Amplification for Mercury Detection.

Mercury (Hg), as a highly harmful environmental pollutant, poses severe ecological and health risks even at low concentrations. Accurate and sensitive methods for detecting Hg2+ ions in aquatic environments are highly needed. In this work, we developed a highly sensitive fluorescence sensor for Hg2+ detection with an integrated use of biosynthetic CdSe/CdS quantum dots (QDs) and liposome carrier signal amplification. To construct such a sensor, three single-stranded DNA probes were rationally designed based on the thymine-Hg2+-thymine (T-Hg2+-T) coordination chemical principles and by taking advantage of the biocompatibility and facile-modification properties of the biosynthetic QDs. Hg2+ could be determined in a range from 0.25 to 100 nM with a detection limit of 0.01 nM, which met the requirements of environmental sample detection. The sensor also exhibited a high selectivity for Hg2+ detection in the presence of other high-level metal ions. A satisfactory capacity of the sensor for detecting environmental samples including tap water, river water, and landfill leachate was also demonstrated. This work opens up a new application scenario for biosynthetic QDs and holds a great potential for environmental monitoring applications.

Factors associated with health-related quality of life in papillary thyroid microcarcinoma patients undergoing radiofrequency ablation: a cross-sectional prevalence study.

PURPOSE: To explore the association of demographic characteristics, clinical symptoms and the fear of the disease progression factors with the physical and mental summary components of the health-related of life (HRQoL) of the papillary thyroid microcarcinoma (PTMC) patients undergoing radiofrequency ablation (RFA). METHODS: 123 PTMC survivors undergoing RFA were enrolled in this study from October 2019 to March 2020. Demographic, clinical symptoms and the fear of the disease progression data were collected. SF-36, THYCA-QoL and FoP-Q-SF were used to evaluate the HRQoL of patients, clinical symptoms and the fear of disease progression. A multivariate regression model was performed to evaluate the association between the independent variable and the HRQoL variable. RESULTS: The average self-reported HRQoL score was 81.17 ± 15.48 for the PCS and 73.40 ± 18.03 for the MCS. The multivariate linear regression model shows that the factors related to a poorer PCS were dependent for the female patients, the symptoms of neuromuscular and the throat/mouth, the fear of disease progression; the psychological disorder, symptoms of throat/mouth, inability to concentrate were related to worse scores for the MCS. The condition that was most strongly related to a poorer HRQoL (in both PCS and MCS) was the fear of their physical health. CONCLUSIONS: The factors related to significantly worse HRQoL scores across PCS and MCS for PTMC survivors include the female gender, the symptoms of neuromuscular and the throat/mouth, the psychological disorder, inability to concentrate, and the fear of their own physical health. Identification, management, and prevention of these factors are critical to improving the HRQoL of patients.

Regeneration of the Neocondyle After Free Fibular Flap Reconstruction of the Mandibular Condyle.

PURPOSE: Shifting of the flap position after condylar reconstruction with free fibular flaps is known to occur, but its long-term effects on postoperative esthetic outcomes have not been sufficiently reported. Therefore, in this study, we evaluated the long-term morphologic stability of the free fibular flap neocondyle. PATIENTS AND METHODS: This was a retrospective cohort study. The primary outcome variables were neocondyle regeneration and neocondyle position including the distance between the glenoid fossa and the initial neocondyle (Fo-Co), the distance between the glenoid fossa and the stable neocondyle (Fo-Co'), and shifting of the neocondyle (defined as the distance between the stable neocondyle and the initial neocondyle). The primary predictor variable was time. The other variables were age, gender, diagnosis, and number of fibular segments. Correlation analysis between the predictor variables and outcome variables was performed. RESULTS: The sample was composed of 26 patients (11 male and 15 female patients) with a mean age of 31 years. Diagnosis and number of fibular segments were significantly associated with Fo-Co and Fo-Co' (P < .05). Among the 26 patients, only 11 showed neocondyle regeneration at follow-up (group A) whereas 15 did not (group B). Neocondyle regeneration was significantly associated with patient age (P < .01). Stable Fo-Co and stable time were significantly associated with neocondyle regeneration (P < .05). The mean stable time was significantly shorter in group A (3.64 ± 1.12 months) than in group B (6.67 ± 3.85 months) (P < .05), and the mean Fo-Co' was significantly shorter in group A (13.65 ± 3.94 mm) than in group B (20.68 ± 8.87 mm) (P < .05). CONCLUSIONS: The possibility of neocondyle regeneration is higher in pediatric patients than in adults. Neocondyle regeneration could result in the movement of the neocondyle toward the glenoid fossa with a shorter stable time, which could improve neocondyle repositioning. Repositioning of the neocondyle with free fibular flaps for mandibular condyle defects is a self-adaption process for temporomandibular joint function.

Double-Barrel Fibula Flap Versus Vascularized Iliac Crest Flap for Mandibular Reconstruction.

PURPOSE: The double-barrel fibula flap and vascularized iliac crest flap are both commonly used for mandibular reconstruction. The present study compared the usage and reconstruction outcomes of transplanted bone with these 2 methods. PATIENTS AND METHODS: The data from 30 patients who had undergone mandibular osteotomy and reconstruction were retrospectively reviewed. Of the 30 patients, 20 received a vascularized iliac crest flap (group A) and 10 received a double-barrel fibula flap (group B). The following variables were compared between the 2 groups: volume of bone flap (VBF), volume of effective bone flap (VEBF; ie, overlap between the volume of the ideal mandible [VIM] and the VBF), usage of the bone flap (VEBF divided by the VBF), mandibular reconstruction rate (VEBF divided by the VIM), volume of needless bone flap (VNBF; ie, VBF minus VEBF; the VNBF included the volume of needless buccal bone flap [VNBBF] and the volume of needless lingual bone flap [VNLBF]), percentage of alveolar crest restoration (PACR; ie, effective bone flap width divided by ideal alveolar crest width), and height of the bone flap (HBF). The independent-samples t test and the χ2 test were used to compare the variables between the 2 groups. Statistical significance was at P ≤ .05. RESULTS: Usage of the bone flap and the length of the mandibular defect were significantly greater in group B than in group A (P = .039 and P < .001, respectively). The VBF, VNBF, and VNLBF were significantly greater in group A than in group B (P < .001 for both). The mandibular reconstruction rate, VNBBF, PACR, HBF, and tooth implantation rate were comparable between the 2 groups. CONCLUSIONS: The double-barrel fibula flap can effectively restore the height of the alveolar crest, reconstruct longer mandibular defects, and provide a better buccal and lingual appearance compared with the vascularized iliac crest flap. Although the vascularized iliac crest flap can provide sufficient bone quantity, it must be contoured to the mandible.

Selection of Guiding Plate Combined With Surgical Navigation for Microsurgical Mandibular Reconstruction.

PURPOSE: The present study summarized selection of guiding plate combined with surgical navigation for microsurgical mandibular reconstruction. METHODS: Data from preoperative maxillofacial enhanced computed tomography (CT) scans were imported to ProPlan CMF. The authors performed virtual mandibulectomy and superimposed 3-dimensional (3D) iliac images on mandibular defects. Guiding plates including mandibular fixation device, reconstruction plate, guiding model, and occlusal splint for various mandibular hemimandibular central lateral (HCL) defects were fabricated to fix bilateral residual mandible. The model was scanned, and data were imported into ProPlan CMF and the intraoperative navigation system. Through landmark points upon the guiding plate, position of the residual mandible was determined during surgical navigation. Intraoperative navigation was used to implement the virtual plan. Sagittal, coronal, axial, and 3D reconstruction images displayed by the navigation system were used to accurately determine osteotomy sites and osteotomy trajectory during surgery. Surgical probe guidance was used to mark the osteotomy line and transfer the virtual procedure to real-time surgery. Accuracy was evaluated using chromatographic analysis. RESULTS: Different guiding plates combined with surgical navigation could be used for various mandibular defects, including mandibular fixation devices for LCL defects, reconstruction plates for LC/L/C defects, and guiding models and occlusal splints for H/L/LC defects (including mandibular ramus). In our study, average and largest shift of the mandible and osteotomy site was <5 mm. CONCLUSION: The authors summarized different ways of combining guiding plates with surgical navigation for reconstruction of various mandibular defects, which could improve clinical outcomes of this procedure with high accuracy.

Effects of Er:YAG laser pre-treatment on dentin structure and bonding strength of primary teeth: an in vitro study.

BACKGROUND: To investigate the effects of Er:YAG laser pre-treatment on the dentin structure and shear bond strength of primary teeth. METHODS: Dentin specimens were prepared using freshly extracted intact primary molars and divided randomly into four groups based on the surface treatment applied. The control and etchant groups received no treatment and conventional acid etching treatment, respectively, while the energy and frequency groups received laser surface treatment with variable energy (50-300 mJ) and frequency (5-30 Hz) parameters. The morphology was observed using scanning electron microscopy. The surface-treated dentin slices were bonded to resin tablets, followed by thermocycle treatment. The shear strength was determined using a universal testing machine and de-bonded surfaces were observed using a stereomicroscope. RESULTS: SEM observation showed that the surface morphology of the dentin slices changed after etching as well as after Er:YAG laser pre-treatment with different energy and frequency values. The dentin tubules opened within a specific energy (50-200 mJ) and frequency (5-20 Hz) range. Beyond this range, the intertubular dentin showed cracks and structural disintegration. Shear strength tests showed no significant changes after acid etching. The shear strength increased significantly (P < 0.05) after Er:YAG laser pre-treatment compared with that of the control group. The shear strength increased within the same energy (50-200 mJ) and frequency (5-20 Hz) range as the tubule opening, but not significantly (P > 0.05). The most common mode of interface failure was adhesive (interface) failure, followed by mixed and resin cohesive failure. CONCLUSIONS: Pre-treatment using Er:YAG laser opens the dentinal tubules without the formation of a smear layer and improves the bonding strength between the primary teeth dentin and the resin composites.

[Factors Affecting Quality of Life of Patients Undergoing Radiofrequency Ablation for Papillary Thyroid Microcarcinoma].

Objective To evaluate the factors affecting health-related quality of life (HRQoL) of patients with thyroid papillary microcarcinoma (PTMC) after ultrasound-guided radiofrequency ablation (RFA).Methods The clinical data of 100 patients with PTMC who underwent reexamination after RFA in the Ultrasound Department of our center from October to December 2019 were retrospectively analyzed.Demographic information was collected.SF-36 and Thyroid Cancer-specific Health-related Quality of Life Questionnaire scales were used to assess patients' quality of life and thyroid-related specific symptoms.The SF-36 scale includes two general domains including physical component summary (PCS) and mental component summary (MCS).The impacts of demographic characteristics and thyroid-related symptoms after RFA on PCS and MCS scores were further analyzed.Results Univariate analysis and correlation analysis showed that the PCS scores in quality of life of PTMC patients were related to sex, neuromuscular, voice, concentration, sympathetic nerve, and throat/mouth complaints, psychological state, sensory symptoms, scar, chills, tingling, and headache (all P<0.1);and the MCS scores were associated with education level, residence, neuromuscular, voice, concentration, sympathetic nerve, and throat/mouth complains, psychological state, sensory symptoms, scar, chills, tingling, and headache (all P<0.1).Multivariate regression analysis showed that the PCS scores were only associated with sex and the neuromuscular and throat/mouth complains and the psychological state.The regression equation was:PCS=110.367-8.025×sex-0.213×psychological state-0.280×neuromuscular complain-0.278×throat/mouth complain.In contrast, the MCS scores were only associated with the psychological state and the throat/mouth and concentration complains, with the regression equation being:MCS=91.323-0.237×psychological state-0.437×throat/mouth-0.304×concentration.Conclusions The main risk factors affecting the quality of life of PTMC patients after ultrasound-guided RFA were female gender, psychological burden, lack of attention, and symptoms in neuromuscular system and throat/mouth.Therefore, preoperative explanations should be made according to the relevant symptoms that the patients may report, and psychological interventions should be offered after RFA to improve the quality of life of PTMC patients after treatment.

10-Year Results of Therapeutic Ratio by Intensity-Modulated Radiotherapy Versus Two-Dimensional Radiotherapy in Patients with Nasopharyngeal Carcinoma.

BACKGROUND: The purpose of this study was to verify 10-year results of survival and late toxicities and assess the ultimate therapeutic ratio of intensity-modulated radiotherapy (IMRT) versus two-dimensional radiotherapy (2DRT) in patients with nasopharyngeal carcinoma (NPC). MATERIALS AND METHODS: We retrospectively reviewed the data from 1,276 patients with nonmetastatic NPC who received IMRT or 2DRT from January 2003 to December 2006. RESULTS: Of the 1,276 patients, 512 were treated with IMRT and 764 with 2DRT. Median follow-up was 115 months. At 10 years, the IMRT group demonstrated significantly better results than the 2DRT group in local failure-free survival (L-FFS; 90% vs. 84%; hazard ratio [HR], 0.57, 95% confidence interval [CI], 0.40-0.81; p = .001), failure-free survival (FFS; 69% vs. 58%; HR, 0.69, 95% CI, 0.57-0.83; p < .001), and overall survival (OS; 75% vs. 63%; HR, 0.62, 95% CI, 0.51-0.77; p < .001). Subgroup multivariate analyses showed that radiotherapeutic technique (IMRT vs. 2DRT) remained an independent prognostic factor for L-FFS in the T1 subgroup (HR, 0.30; 95% CI, 0.11-0.80; p = .02); for FFS in the stage II subgroup (HR, 0.42; 95% CI, 0.24-0.73; p = .002); and for OS in the stage I (HR, 0.20; 95% CI, 0.04-0.96; p = .04), stage II (HR, 0.39; 95% CI, 0.21-0.75; p = .004), and stage IVA-B (HR, 0.74, 95% CI, 0.56-0.98; p = .04) subgroups. The incidence of grade 3-4 temporal lobe necrosis, cranial neuropathy, eye damage, ear damage, neck soft tissue damage, trismus, and dry mouth was significantly lower in the IMRT group than in the 2DRT group. CONCLUSION: IMRT demonstrated an improved ultimate therapeutic ratio compared with 2DRT in patients with NPC after a 10-year follow-up, with significant improvement of L-FFS, FFS, and OS and decrease in most late toxicities. IMPLICATIONS FOR PRACTICE: The ultimate therapeutic ratio of intensity-modulated radiotherapy versus two-dimensional radiotherapy in patients with nasopharyngeal carcinoma is unclear. In this retrospective study of 1,276 patients with nonmetastatic nasopharyngeal carcinoma with a follow-up of 115 months, intensity-modulated radiotherapy demonstrated an improved ultimate therapeutic ratio compared with two-dimensional radiotherapy, with significant improvement of local failure-free survival, failure-free survival, and overall survival and decrease in most late toxicities and noncancer deaths. However, distant control remains insufficient with this treatment modality.

Highly Selective Capture of Monophosphopeptides by Two-Dimensional Metal-Organic Framework Nanosheets.

Separation of monophosphopeptides from multi-phosphopeptides in complex biological samples is significant in the study of protein kinase signal transduction pathways. To the best of our knowledge, very few materials have been reported that could selectively enrich monophosphopeptides because of the chemical difficulty in retaining the intermediate monophosphopeptides and excluding both non-phosphopeptides and multi-phosphopeptides in acidic conditions, which requires unique interactions to balance the metallic affinity and the hydrophobicity. With the large surface area, abundant accessible active sites, and ultrathin structures, two-dimensional (2-D) metal-organic framework (MOF) Hf-1,3,5-tris(4-carboxyphenyl)benzene (BTB) nanosheets were rationally selected. Due to the elongated organic ligands and the balance between metallic affinity of clusters and hydrophobicity from ligands, the 2-D Hf-BTB nanosheets exhibited unique enrichment selectivity toward monophosphopeptides. The 2-D MOF nanosheets demonstrated excellent sensitivity (detection limit of 0.4 fmol µL-1) and selectivity [1:1000 molar ratios of ß-casein/BSA (bovine serum albumin)] in model phosphopeptides enrichment. The nanosheets were implemented for the analysis of nonfat milk and human saliva samples as well as in situ isotope labeling for dysregulated phosphopeptides from patients' serum with anal canal inflammation, exhibiting 6.6-fold upregulation of serum phosphopeptide HS4 (ADpSGEGDFLAEGGGVR) compared to the control healthy serum. The proteomics analysis of mouse brain cortical samples associated with Alzheimer's disease, which were from Akt (protein kinase B) conditional knockout mouse and littermate control mouse, was further established with 2-D Hf-BTB nanosheets. With high capture efficiency for monophosphopeptides, this method was capable of distinguishing the difference of monophosphopeptides from microtubule-associated protein τ (MAPT/τ) between the Akt knockout sample and control sample.

Abrogation of Fam20c altered cell behaviors and BMP signaling of immortalized dental mesenchymal cells.

FAM20C mutations compromise the mineralization of skeleton and tooth in both human and mouse. Putatively, the mineralization disorder is attributed to the elevated fibroblast growth factor 23 (FGF23), which reduced the serum phosphorus by suppressing the reabsorption of phosphorus in kidney. Besides the regulation on systemic phosphorus homeostasis, FAM20C was also implicated to regulate cell behaviors and gene expression through a cell-autonomous manner. To identify the primary effects of Fam20c on dental mesenchymal cells, mouse Fam20c-deficient dental mesenchymal cells were generated by removing the floxed alleles from the immortalized mouse Fam20cf/f dental mesenchymal cells with Cre-expressing lentivirus. The removal of Fam20c exerted no impact on cell morphology, but suppressed the proliferation and mobility of the dental mesenchymal cells. Fam20c deficiency also significantly reduced the expression of Osterix, Runx2, type I Collagen a 1 (Col1a1), Alkaline phosphatase (Alpl) and the members of the small integrin-binding ligand, N-linked glycoprotein (SIBLING) family, but increased Fgf23 expression. Consistently, the in vitro mineralization of Fam20c-deficient dental mesenchymal cells was severely disabled. However, supplements of the non-collagenous proteins from wild type rat dentin failed to rescue the compromised mineralization, suggesting that the roles of FAM20C in tooth mineralization are more than phosphorylating local matrices and regulating systemic phosphorus metabolism. Moreover, the down-regulated BMP signaling pathways in the Fam20c deficient dental mesenchymal cells revealed that the kinase activity of FAM20C might be required to maintain BMP signaling. In summary, our study discloses that Fam20c indeed regulates cell behaviors and cell signaling pathway in a cell-autonomous manner.

FAM20C regulates osteoblast behaviors and intracellular signaling pathways in a cell-autonomous manner.

Recent studies indicate that Family with sequence similarity 20 member C (FAM20C) catalyzes the phosphorylation of secreted proteins, and participates in a variety of biological processes, including cell proliferation, migration, mineralization, and phosphate homeostasis. To explore the local influences of FAM20C on osteoblast, Fam20c-deficient osteoblasts were generated by treating the immortalized Fam20cf/f osteoblasts with CMV-Cre-IRES-EGFP lentivirus. Compared with the normal Fam20cf/f osteoblasts, the expression of Bone sialoprotein (Bsp), Osteocalcin (Ocn), Fibroblast growth factor 23 (Fgf23), and transcription factors that promote osteoblast maturation were up-regulated in the Fam20c-deficient osteoblasts. In contrast, the expression of Dental matrix protein 1 (Dmp1), Dentin sialophosphoprotein (Dspp), Osteopontin (Opn), type I Collagen a 1 (Col1a1), and Alkine phosphatase (Alp) were down-regulated in the Fam20c-deficient cells. These alterations disclosed the primary regulation of Fam20c on gene expression. The Fam20c-deficient osteoblasts showed a remarkable reduction in the ability of forming mineralized nodules. However, supplements of extracellular matrix proteins extracted from the normal bone failed to rescue the reduced mineralization, suggesting that FAM20C may affect the biomineralization by the means more than local phosphorylation of extracellular matrix proteins and systemic phosphorus homeostasis. Moreover, although Fam20c deficiency had little impact on cell proliferation, it significantly reduced cell migration and lowered the levels of p-Smad1/5/8, p-Erk and p-p38, suggesting that the kinase activity of FAM20C might be essential to cell mobility and the activity of BMP ligands. In summary, these findings provide evidences that FAM20C may regulate osteoblast maturation, migration, mineralization, and BMP signaling pathways in a cell-autonomous manner.

Two New Preyssler-Type Polyoxometalate-Based Coordination Polymers and Their Application in Horseradish Peroxidase Immobilization.

Enzyme immobilization is of increasing importance for biocatalysis, for which good supports are critical. Herein, two new Preyssler-type polyoxometalate (POM)-based coordination polymers, namely, {[Cu(H2 biim)2 ][{Cu(H2 biim)2 (µ-H2 O)}2 Cu(H2 biim)(H2 O)2 ]H[({Cu(H2 biim)(H2 O)2 }0.5 )2 ((µ-C3 HN2 Cl2 ){Cu(H2 biim)}2 ){Z(H2 O)P5 W30 O110 }]⋅x H2 O}n (1: Z=Na, x=9; 2: Z=Ag, x=10; H2 biim=2,2'-biimidazole) were designed and synthesized. Compounds 1 and 2 exhibit the same skeletons, which contain multiple CuII complex fragments and penta-supported {ZP5 W30 } (Z=Na, Ag) clusters. They were first employed to immobilize horseradish peroxidase (HRP). Results show that compounds 1 and 2 are good supports for HRP immobilization, and exhibit higher enzyme loading, lower loading times, and excellent reusability. The immobilized HRP (HRP/1 or HRP/2) was further applied to detect H2 O2 , and good sensitivity, wide linear range, low detection limit, and fast response were achieved. This work shows that POM-based hybrid materials are a new kind of promising support for enzyme immobilization.

The Predictive Value of Baseline HBsAg Level and Early Response for HBsAg Loss in Patients with HBeAg-positive Chronic Hepatitis B during Pegylated Interferon Alpha-2a Treatment.

OBJECTIVE: To explore the predictive value of baseline HBsAg level and early response for HBsAg loss in patients with HBeAg-positive chronic hepatitis B during pegylated interferon alpha-2a treatment. METHODS: A total of 121 patients with HBeAg-positive chronic hepatitis B who achieved HBsAg loss were enrolled; all patients were treated with PEG-IFNα-2a 180 µg/week. Serum HBV DNA and serological indicators (HBsAg, anti-HBs, HBeAg, and anti-HBe) were determined before and every 3 months during treatment. RESULTS: The median treatment time for HBsAg loss was 84 weeks (7-273 weeks), and 74.38% (90 cases) of the patients needed extended treatment (> 48 weeks). The correlation between baseline HBsAg levels and the treatment time of HBsAg loss was significant (B = 14.465, t = 2.342, P = 0.021). Baseline HBsAg levels together with the decline range of HBsAg at 24 weeks significantly correlated with the treatment time of HBsAg loss (B = 29.862, t = 4.890, P = 0.000 and B = 27.993, t = 27.993, P = 0.005). CONCLUSION: Baseline HBsAg levels and extended therapy are critical steps toward HBsAg loss. Baseline HBsAg levels together with early response determined the treatment time of HBsAg loss in patients with HBeAg-positive chronic hepatitis B during pegylated interferon alpha-2a treatment.