Mir155hg Accelerates Hippocampal Neuron Injury in Convulsive Status Epilepticus by Inhibiting Microglial Phagocytosis.

Convulsive status epilepticus (CSE) is a common critical neurological condition that can lead to irreversible hippocampal neuron damage and cognitive dysfunction. Multiple studies have demonstrated the critical roles that long non-coding RNA Mir155hg plays in a variety of diseases. However, less is known about the function and mechanism of Mir155hg in CSE. Here we investigate and elucidate the mechanism underlying the contribution of Mir155hg to CSE-induced hippocampal neuron injury. By applying high-throughput sequencing, we examined the expression of differentially expressed genes in normal and CSE rats. Subsequent RT-qPCR enabled us to measure the level of Mir155hg in rat hippocampal tissue. Targeted knockdown of Mir155hg was achieved by the AAV9 virus. Additionally, we utilized HE and Tunel staining to evaluate neuronal injury. Immunofluorescence (IF), Golgi staining, and brain path clamping were also used to detect the synaptic plasticity of hippocampal neurons. Finally, through IF staining and Sholl analysis, we assessed the degree of microglial phagocytic function. It was found that the expression of Mir155hg was elevated in CSE rats. HE and Tunel staining results showed that Mir155hg knockdown suppressed the hippocampal neuron loss and apoptosis followed CSE. IF, Golgi staining and brain path clamp data found that Mir155hg knockdown enhanced neuronal synaptic plasticity. The results from IF staining and Sholl analysis showed that Mir155hg knockdown enhanced microglial phagocytosis. Our findings suggest that Mir155hg promotes CSE-induced hippocampal neuron injury by inhibiting microglial phagocytosis.

Protective effect of alirocumab, a PCSK9 inhibitor, on the sciatic nerve of rats with diabetic peripheral neuropathy.

Dyslipidemia has been considered a risk factor for diabetic peripheral neuropathy. Proprotein convertase subtilisin-like/Kexin 9 inhibitor (PCSK9) inhibitors are a new type of lipid-lowering drug currently in clinical use. The role of PCSK9 in diabetic peripheral neuropathy is still unclear. In this study, the effect of alirocumab, a PCSK9 inhibitor, on the sciatic nerve in rats with diabetic peripheral neuropathy and its underlying mechanisms were investigated. The diabetic peripheral neuropathy rat model was established by using a high-fat diet combined with streptozotocin injection, and experimental subjects were divided into normal, diabetic peripheral neuropathy, and alirocumab groups. The results showed that Alirocumab improved nerve conduction, morphological changes, and small fiber deficits in rats with DPN, possibly related to its amelioration of oxidative stress and the inflammatory response.

LncRNA H19 Regulates P-glycoprotein Expression Through the NF-κB Signaling Pathway in the Model of Status Epilepticus.

Pharmaco-resistance is a challenging problem for treatment of status epilepticus (SE) in the clinic. P-glycoprotein (P-gp) is one of the most important multi-drug transporters that contribute to drug resistance of SE. Long noncoding RNAs (lncRNAs) have been increasingly recognized as versatile regulators of P-gp in tumors and epilepsy. However, the function of lncRNAs in drug resistance of SE remains largely unknown. In the present study, pilocarpine-induced rat model is used to explore the expression profiles of lncRNAs in the hippocampus of SE using RNA sequencing. Our results implied that the level of lncRNA H19 was significantly increased in the hippocampus of SE rats, which was positively correlated with the level of P-gp. While downregulation of H19 could inhibit the expression of P-gp and alleviate neural damage in the hippocampus of SE rats. Furthermore, it was revealed that H19 regulates P-gp expression through the nuclear factor-kappaB (NF-κB) signaling pathway by functioning as a competing endogenous RNA against microRNA-29a-3p. Overall, our study indicated that H19 regulates P-gp expression and neural damage induced by SE through the NF-κB signaling pathway, which provides a promising target to overcome drug resistance and alleviate brain damage for SE.

Perivascular fat attenuation index value and plaque volume increased in non-target lesions of coronary arteries after stenting.

BACKGROUND: Progression of non-target lesions (NTLs) after stenting has been reported and is associated with the triggering of an inflammatory response. The perivascular fat attenuation index (FAI) may be used as a novel imaging biomarker for the direct quantification of coronary inflammation. OBJECTIVES: To investigate whether FAI values can help identify changes in inflammation status in patients undergoing stent implantation, especially in NTLs. METHODS: Patients who underwent pre- and post-stenting coronary computed tomography angiography (CCTA) examination between January 2015 and February 2021 were consecutively enrolled. The pre- and post-stenting FAIs of the full coronary arteries were compared in both the non- and stent-implanted coronary arteries. Moreover, local FAI values were measured and compared between the NTLs and target lesions in the stent implantations. We also compared changes in plaque type and volume in NTLs before and after stenting. RESULTS: A total of 89 patients (mean age 61 years; male 59) were enrolled. The perivascular FAI values in the full coronary arteries decreased after stenting in both the non- and stent-implanted coronary arteries, similar to those in the target lesions. Conversely, the perivascular FAI values in the NTLs increased after stenting (p < 0.05). In addition, the plaque volumes significantly increased in the NTLs after stenting, regardless of whether they were non-calcified, mixed, or calcified (p < 0.05). CONCLUSION: Perivascular FAI values and plaque volumes increased in the NTLs after stenting. Perivascular FAI can be a promising imaging biomarker for monitoring coronary inflammation after stenting and facilitate long-term monitoring in clinical settings. CLINICAL RELEVANCE STATEMENT: Perivascular fat attenuation index, a non-invasive imaging biomarker, may help identify coronary arteries with high inflammation in non-target lesions and facilitate long-term monitoring, potentially providing an opportunity for more targeted treatment. KEY POINTS: • Perivascular fat attenuation index (FAI) values and plaque volumes increased in the non-target lesions (NTLs) after stenting, suggesting potential focal inflammation progression after stenting. However, stenting along with anti-inflammatory treatment ameliorated inflammation in the full coronary arteries. • Perivascular FAI, a non-invasive imaging biomarker, may help identify coronary arteries with high inflammation in NTLs and facilitate long-term monitoring, potentially providing an opportunity for more targeted treatment.

Single Small Extracellular Vesicle (sEV) Quantification by Upconversion Nanoparticles.

Cancer-derived small extracellular vesicles (sEVs) are potential circulating biomarkers in liquid biopsies. However, their small sizes, low abundance, and heterogeneity in molecular makeups pose major technical challenges for detecting and characterizing them quantitatively. Here, we demonstrate a single-sEV enumeration platform using lanthanide-doped upconversion nanoparticles (UCNPs). Taking advantage of the unique optical properties of UCNPs and the background-eliminating property of total internal reflection fluorescence (TIRF) imaging technique, a single-sEV assay recorded a limit of detection 1.8 × 106 EVs/mL, which was nearly 3 orders of magnitude lower than the standard enzyme-linked immunosorbent assay (ELISA). Its specificity was validated by the difference between EpCAM-positive and EpCAM-negative sEVs. The accuracy of the UCNP-based single-sEV assay was benchmarked with immunomagnetic-beads flow cytometry, showing a high correlation (R2> 0.99). The platform is suitable for evaluating the heterogeneous antigen expression of sEV and can be easily adapted for biomarker discoveries and disease diagnosis.

Effect of smile esthetics on the quality of life in a Han Chinese population.

OBJECTIVE: This study aimed to explore the influence of smile esthetics on the quality of life in a Han Chinese population. METHODS: A total of 110 Han Chinese volunteers (52 males and 58 females, mean age 26.7 years) were recruited, and frontal images of their smiles were collected. A photoediting program was used to evaluate the following components of smile esthetics: the buccal corridor ratios, the angle between the interpupillary and commissural lines, the number of visible teeth, and width-to-length ratios of the maxillary central incisors. Oral health-related quality of life (OHRQoL) in these volunteers was evaluated using the Psychosocial Impact of Dental Aesthetics Questionnaire (PIDAQ) and Oral Health Impact Profile-14 (OHIP-14). Statistical analyses were performed by using Pearson's correlations and multiple linear regression (α = 0.05). RESULTS: Compared to males, females scored higher on psychological impact (p = 0.017). Moreover, the number of visible teeth had a significant effect on social impact, psychological impact and esthetic concern (all p < 0.05). However, the buccal corridor ratios, the angle between the interpupillary and commissural lines, and the width-to-length ratios of the maxillary central incisor did not correlate with the OHRQoL (all p > 0.05). CONCLUSIONS: The number of visible teeth significantly influenced the quality of life in a Han Chinese population. Smile esthetics had a significantly greater psychological impact on females than males. CLINICAL SIGNIFICANCE: This study highlights the effects of smile esthetics on the quality of life in a Han Chinese population. Female patients may raise more esthetic concerns in clinical practice.

Upconversion nanoparticle-assisted single-molecule assay for detecting circulating antigens of aggressive prostate cancer.

Sensitive and quantitative detection of molecular biomarkers is crucial for the early diagnosis of diseases like metabolic syndrome and cancer. Here we present a single-molecule sandwich immunoassay by imaging the number of single nanoparticles to diagnose aggressive prostate cancer. Our assay employed the photo-stable upconversion nanoparticles (UCNPs) as labels to detect the four types of circulating antigens in blood circulation, including glypican-1 (GPC-1), leptin, osteopontin (OPN), and vascular endothelial growth factor (VEGF), as their serum concentrations indicate aggressive prostate cancer. Under a wide-field microscope, a single UCNP doped with thousands of lanthanide ions can emit sufficiently bright anti-Stokes' luminescence to become quantitatively detectable. By counting every single streptavidin-functionalized UCNP which specifically labeled on each sandwich immune complex across multiple fields of views, we achieved the Limit of Detection (LOD) of 0.0123 ng/ml, 0.2711 ng/ml, 0.1238 ng/ml, and 0.0158 ng/ml for GPC-1, leptin, OPN and VEGF, respectively. The serum circulating level of GPC-1, leptin, OPN, and VEGF in a mixture of 10 healthy normal human serum was 25.17 ng/ml, 18.04 ng/ml, 11.34 ng/ml, and 1.55 ng/ml, which was within the assay dynamic detection range for each analyte. Moreover, a 20% increase of GPC-1 and OPN was observed by spiking the normal human serum with recombinant antigens to confirm the accuracy of the assay. We observed no cross-reactivity among the four biomarker analytes, which eliminates the false positives and enhances the detection accuracy. The developed single upconversion nanoparticle-assisted single-molecule assay suggests its potential in clinical usage for prostate cancer detection by monitoring tiny concentration differences in a panel of serum biomarkers.

HMGA2 contributes to vascular development and sprouting angiogenesis by promoting IGFBP2 production.

Angiogenesis is the process by which new blood vessels form from preexisting vessels and regulates the processes of embryonic development, wound healing and tumorigenesis. HMGA2 is involved in the occurrence of several cancers, but its biological role and the exact downstream genes involved in vascular development and sprouting angiogenesis remain largely unknown. Here, we first found that HMGA2 knockdown in zebrafish embryos resulted in defects of central artery formation. RNA sequencing revealed that IGFBP2 was significantly downregulated by interference with HMGA2, and IGFBP2 overexpression reversed the inhibition of brain vascular development caused by HMGA2 deficiency. In vitro, we further found that HMGA2 knockdown blocked the migration, tube formation and branching of HUVECs. Similarly, IGFBP2 protein overexpression attenuated the impairments induced by HMGA2 deficiency. Moreover, the promotion of angiogenesis by HMGA2 overexpression was verified in a Matrigel plug assay. We next found that HMGA2 bound directly to a region in the IGFBP2 promoter and positively regulated IGFBP2 expression. Interestingly, the mRNA expression levels of HMGA2 and IGFBP2 were increased significantly in the peripheral blood of hemangioma patients, indicating that overexpression of HMGA2 and IGFBP2 results in vessel formation, consistent with the results of the in vivo and in vitro experiments. In summary, our findings demonstrate that HMGA2 promotes central artery formation by modulating angiogenesis via IGFBP2 induction.

LncRNA Snhg5 Attenuates Status Epilepticus Induced Inflammation through Regulating NF-κΒ Signaling Pathway.

Status epilepticus (SE) induced inflammation plays an important role in the pathogenesis of SE. Long non-coding RNA small nucleolar RNA host gene 5 (lncRNA Snhg5) has been reported in various inflammatory diseases. However, the mechanism of Snhg5 regulated inflammation in SE remains unclear. Therefore, this study aimed to clarify the role and mechanism of Snhg5 in SE-induced inflammation in vitro and vivo. In vitro, lipopolysaccharide (LPS)-induced inflammation in microglia was used to mimic the inflammation after SE. In vivo, SE model was induced by lithium chloride and pilocarpine. The level of Snhg5, p65, p-p65, p-inhibitor of kappaB (IκB)α, IκBα and inflammatory factors (tumor necrosis factor (TNF)-α, interleukin (IL)-1ß) were measured via quantitative real-time PCR or Western blot. The Nissl stain and immunohistochemical stain were performed to observe hippocampal damage and microglia proliferation. The results showed Snhg5 was up-regulated in the rat and microglia. Knockdown of Snhg5 inhibited LPS-induced inflammation and relative expression of p-65/p65, p-IκBα/IκBα. Moreover, down-regulation of Snhg5 attenuated SE-induced inflammation and reduced the number of microglia in hippocampus. These findings indicated that Snhg5 modulates the inflammation via nuclear factor-kappaB (NF-κB) signaling pathway in SE rats.

Does laser treatment restore the bond strength of resin composites to recently bleached enamel? A systematic review and meta-analysis.

PURPOSE: To do a systematic review and meta-analysis to determine whether laser treatment affects the bond strength of resin composites to recently bleached enamel. METHODS: This report follows the Preferred Reporting Items for Systematic Reviews and Qualitative Analyses (PRISMA) statement. Medline via PubMed, Embase, Web of Science, and the Cochrane Library databases were searched with no limits on publication year. Two reviewers independently screened all titles and abstracts to perform the study selection, data extraction, and risk-of-bias assessments. A random-effects meta-analysis model was performed using Review Manager software (version 5.3, Cochrane Collaboration). RESULTS: From the 93 records identified, seven articles that met all the inclusion criteria were included in the systematic review, and six studies were included in the meta-analysis. The overall results showed a statistically significant difference in bond strength between the control group and laser-treated group (P= 0.04; mean difference: 5.27; 95% confidence interval: 0.28 to 10.27), favoring the laser-treated group. Subgroup analyses revealed that the tooth source (bovine or human teeth) contributed to the effect of laser treatment on the bleached enamel. CLINICAL SIGNIFICANCE: Laser treatment may increase the bond strength of resin composites to recently bleached enamel. Pretreatment with a laser, preferably with Nd:YAG (1 W, frequency of 10 Hz, irradiation time of 60 seconds) or CO2 lasers (0.5 W, frequency of 10 Hz, irradiation time of 60 seconds), may be recommended to restore the bond strength of recently bleached enamel.

Preselectable Optical Fingerprints of Heterogeneous Upconversion Nanoparticles.

The control in optical uniformity of single nanoparticles and tuning their diversity in multiple dimensions, dot to dot, holds the key to unlocking nanoscale applications. Here we report that the entire lifetime profile of the single upconversion nanoparticle (τ2 profile) can be resolved by confocal, wide-field, and super-resolution microscopy techniques. The advances in both spatial and temporal resolutions push the limit of optical multiplexing from microscale to nanoscale. We further demonstrate that the time-domain optical fingerprints can be created by utilizing nanophotonic upconversion schemes, including interfacial energy migration, concentration dependency, energy transfer, and isolation of surface quenchers. We exemplify that three multiple dimensions, including the excitation wavelength, emission color, and τ2 profile, can be built into the nanoscale derivative τ2-dots. Creating a vast library of individually preselectable nanotags opens up a new horizon for diverse applications, spanning from sub-diffraction-limit data storage to high-throughput single-molecule digital assays and super-resolution imaging.

Comparison of smile esthetics among celebrities, dentists, and dental students in a Han Chinese population.

STATEMENT OF PROBLEM: Various components of smile esthetics have been evaluated. However, whether these components affect smile esthetics in Han Chinese celebrities, dentists, and dental students is unknown. PURPOSE: The purpose of this study was to evaluate the smile esthetics of celebrities, dentists, and dental students in a Han Chinese population. MATERIAL AND METHODS: Frontal images of smiles were obtained from the Internet for the celebrity group, while images of dentists and dental students were collected for the dentist and dental student groups. The buccal and posterior corridor ratios, the alignment discrepancy between the commissural and interpupillary lines, the number of teeth revealed, the width ratio relative to the golden proportion, and the width-to-length ratios of maxillary anterior teeth were calculated by using a photo-editing software program. Statistical analysis was performed by using 2-way ANOVA and the Tukey HSD test (α=.05). RESULTS: Eligible images were obtained for 81 celebrities, 83 dentists, and 78 dental students. The celebrities were found to have the highest buccal and posterior corridor ratios and reveal the least teeth in the smile among the 3 groups (all P<.001). The alignment discrepancy for celebrities was significantly less than that for dental students (P=.025). Moreover, the absolute difference of width ratios from the golden proportion was significantly different from 0 for all groups (all P<.001). CONCLUSIONS: Significant differences were found in the smiles of Han Chinese celebrities, dentists, and dental students in regard to the buccal and posterior corridor ratios, the alignment discrepancy between the commissural and interpupillary lines, and the number of teeth revealed. Chinese celebrities tended to have a smile that displays to the second premolar, with a medium buccal corridor width.

Effects of remaining dentin thickness on the bond strength of bleached dentin.

BACKGROUND: The bond strength of resin composites to dentin was reported to be related to either the remaining dentin thickness (RDT) or bleaching treatment. However, information is limited regarding the effects of RDT on the bond strength of bleached dentin. The present study aimed to investigate the effects of RDT on the microshear bond strength (µSBS) of resin cement to bleached dentin. METHODS: A total of 120 dentin specimens were prepared and randomly divided into 2 groups: a bleaching group (group B) and a control group (group C). Hydrogen peroxide with a concentration of 35% (Ultradent, USA) was applied on the dentin surface for 2 × 1 d for group B, while no bleaching treatment was performed for group C. After the treatment, the specimens were finished and polished to obtain different RDTs (2, 1, and 0.5 mm) and divided into 3 groups of 20 specimens each. The bonding procedure was performed using Panavia V5 (Kuraray, Japan) with a bonding area of 0.785 mm2. For each group, half of the specimens were subjected to 5000 thermal cycles (subgroup T), while the other half did not receive thermocycling (subgroup N) (n = 10). The specimens were then subjected to the µSBS test using a universal testing machine. Data were analyzed by a three-way analysis of variance (α = 0.05). The fracture modes of the specimens were confirmed with a measuring microscope. Representative specimens with different fracture modes were observed with scanning electron microscopy (SEM). RESULTS: The µSBS values were significantly affected by bleaching treatment (p < 0.001), whereas no significant effect was observed for thermocycling (p = 0.293). In terms of RDT, a significantly different µSBS value was found among the subgroups with different RDTs in group C (p = 0.003). However, the RDT did not significantly affect the µSBS values of bleached dentin in group B (p = 0.779). The µSBS values were significantly lower in group B than in group C (p < 0.001). A higher percentage of adhesive failure was observed in group B than in group C. CONCLUSION: Based on the present findings, it can be concluded that the RDT did not affect the bond strength of resin cement to bleached dentin. CLINICAL SIGNIFICANCE: Since RDT did not affect the bond strength of resin cement to bleached dentin, bonding procedures should not be performed immediately after intracoronal bleaching, even if the dentin is planned to be removed due to a tooth preparation process.

Facile manipulation of protein localization in fission yeast through binding of GFP-binding protein to GFP.

GFP-binding protein (or GBP) has been recently developed in various systems and organisms as an efficient tool to purify GFP-fusion proteins. Due to the high affinity between GBP and GFP or GFP variants, this GBP-based approach is also ideally suited to alter the localization of functional proteins in live cells. In order to facilitate the wide use of the GBP-targeting approach in the fission yeast Schizosaccharomyces pombe, we developed a set of pFA6a-, pJK148- and pUC119-based vectors containing GBP- or GBP-mCherry-coding sequences and variants of inducible nmt1 or constitutive adh1 promoters that result in different levels of expression. The GBP or GBP-mCherry fragments can serve as cassettes for N- or C-terminal genomic tagging of genes of interest. We illustrated the application of these vectors in the construction of yeast strains with Dma1 or Cdc7 tagged with GBP-mCherry and efficient targeting of Dma1- or Cdc7-GBP-mCherry to the spindle pole body by Sid4-GFP. This series of vectors should help to facilitate the application of the GBP-targeting approach in manipulating protein localization and the analysis of gene function in fission yeast, at the level of single genes, as well as at a systematic scale.

MicroRNA-146a-5p Downregulates the Expression of P-Glycoprotein in Rats with Lithium-Pilocarpine-Induced Status Epilepticus.

Increasing evidence supports that the efflux transporters, especially P-glycoprotein (P-gp), have vital roles on drug resistance in epilepsy. Overexpression of P-gp in the brain could reduce the anti-epileptic drugs (AEDs) concentration in the epileptogenic zone, resulting in drug resistance. Studies have demonstrated that recurrent seizures induce the expression of P-gp and status epilepticus (SE) could upregulate the expression of P-gp, resulting in drug resistance. MicroRNAs (miRNAs), as endogenous regulators, represent small regulatory RNA molecules that have been shown to act as negative regulators of gene expression in different biological processes. We investigated the impact of miR-146a-5p on the expression of P-gp in status epilepticus rat model. The expression of miR-146a-5p in rat cortex and hippocampus was measured by quantitative RT-PCR at 2 weeks after induction of SE. Meanwhile, we detected the expression of P-gp in the brain of SE rats using Western blotting and immunohistochemistry. Upregulation of miR-146a-5p and overexpression of P-gp were evident at 2 weeks after SE. Moreover, the expression of P-gp was downregulated by injection of miR-146a mimic into the hippocampus. We also detected the expression of interleukin-1 receptor-associated protein kinases-1 (IRAK1) and tumor necrosis factor receptor-associated factor 6 (TRAF6) and nuclear factor-kappaB (NF-κB) p65 using Western blotting and immunohistochemistry, which indicated the expression of IRAK1, TRAF6 and NF-κB p-p65/p65 increased in the brain of SE rats, and overexpression of miR-146a-5p could downregulate the expression of IRAK1, TRAF6, NF-κB p-p65/p65 and P-gp. Our study indicated that miR-146a-5p may decrease the expression of P-gp in status epilepticus rats via NF-κB signaling pathway.

Nanoparticle-microRNA-146a-5p polyplexes ameliorate diabetic peripheral neuropathy by modulating inflammation and apoptosis.

Nontoxic and nonimmunogenic nanoparticles play an increasingly important role in the application of pharmaceutical nanocarriers. The pathogenesis of diabetic peripheral neuropathy (DPN) has been extensively studied. However, the role of microRNAs in DPN remains to be clarified. We verified in vitro that miR-146a-5p mimics inhibited the expression of proinflammatory cytokines and apoptosis. Then, we explored the protective effect of nanoparticle-miRNA-146a-5p polyplexes (nano-miR-146a-5p) on DPN rats. We demonstrated that nano-miR-146a-5p improved nerve conduction velocity and alleviated the morphological damage and demyelination of the sciatic nerve of DPN rats. The expression of the inflammatory cytokines, caspase-3, and cleaved caspase-3 in the sciatic nerve was inhibited by nano-miR-146a-5p. Additionally, nano-miR-146a-5p increased the expression of myelin basic protein. These results all indicated that nano-miR-146a-5p had a protective effect on peripheral nerves in the DPN rat model, which may occur through the regulation of the inflammatory response and apoptosis.

Finish-line designs for ceramic crowns: A systematic review and meta-analysis.

STATEMENT OF PROBLEM: Different finish-line designs have been advocated for tooth preparations of ceramic crowns. However, scientific evidence is lacking to help clinicians make a proper selection. PURPOSE: The purpose of this systematic review and meta-analysis was to evaluate the effects of finish-line designs on the marginal and internal adaptations of ceramic crowns. MATERIAL AND METHODS: This report follows the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) statement. The literature search was conducted in MEDLINE via the PubMed, Embase, and Web of Science databases with no publication year or language limits. In vitro studies comparing the marginal and internal adaptations of ceramic crowns with rounded shoulder and chamfer finish lines were included in the meta-analysis. RESULTS: Sixteen studies were included in the qualitative synthesis and meta-analyses. Statistical analyses were conducted using the Review Manager Software. Meta-analyses were performed with random-effects models (α=.05). Ceramic crowns with rounded shoulders exhibited significantly better marginal adaptation than those with chamfers (P<.001; mean difference=-7.8; 95% confidence interval=-11.6 to -4.1). Moreover, ceramic crowns with chamfers exhibited significantly better internal adaptation than those with rounded shoulders (P=.020; mean difference=35.0; 95% confidence interval=6.5 to 63.5). CONCLUSIONS: The difference in marginal adaptation of ceramic crowns using 2 finish-line designs was small, and the clinical significance was low, whereas the results of internal adaptation favored the chamfer finish line.

Bispecific Antibody-Functionalized Upconversion Nanoprobe.

Upconversion nanoparticles (UCNPs) are new optical probes for biological applications. For specific biomolecular recognition to be realized for diagnosis and imaging, the key lies in developing a stable and easy-to-use bioconjugation method for antibody modification. Current methods are not yet satisfactory regarding conjugation time, stability, and binding efficiency. Here, we report a facile and high-yield approach based on a bispecific antibody (BsAb) free of chemical reaction steps. One end of the BsAb is designed to recognize methoxy polyethylene glycol-coated UCNPs, and the other end of the BsAb is designed to recognize the cancer antigen biomarker. Through simple vortexing, BsAb-UCNP nanoprobes form within 30 min and show higher (up to 54%) association to the target than that of the traditional UCNP nanoprobes in the ELISA-like assay. We further demonstrate its successful binding to the cancer cells with high efficiency and specificity for background-free fluorescence imaging under near-infrared excitation. This method suggests a general approach broadly suitable for functionalizing a range of nanoparticles to specifically target biomolecules.

Exonuclease III-Assisted Upconversion Resonance Energy Transfer in a Wash-Free Suspension DNA Assay.

Sensitivity is the key in optical detection of low-abundant analytes, such as circulating RNA or DNA. The enzyme Exonuclease III (Exo III) is a useful tool in this regard; its ability to recycle target DNA molecules results in markedly improved detection sensitivity. Lower limits of detection may be further achieved if the detection background of autofluorescence can be removed. Here we report an ultrasensitive and specific method to quantify trace amounts of DNA analytes in a wash-free suspension assay. In the presence of target DNA, the Exo III recycles the target DNA by selectively digesting the dye-tagged sequence-matched probe DNA strand only, so that the amount of free dye removed from the probe DNA is proportional to the number of target DNAs. Remaining intact probe DNAs are then bound onto upconversion nanoparticles (energy donor), which allows for upconversion luminescence resonance energy transfer (LRET) that can be used to quantify the difference between the free dye and tagged dye (energy acceptor). This scheme simply avoids both autofluorescence under infrared excitation and many tedious washing steps, as the free dye molecules are physically located away from the nanoparticle surface, and as such they remain "dark" in suspension. Compared to alternative approaches requiring enzyme-assisted amplification on the nanoparticle surface, introduction of probe DNAs onto nanoparticles only after DNA hybridization and signal amplification steps effectively avoids steric hindrance. Via this approach, we have achieved a detection limit of 15 pM in LRET assays of human immunodeficiency viral DNA.

Medical Applications of Collagen and Hyaluronan in Regenerative Medicine.

In order to develop and commercialize for the regenerative medicinal products, smart biomaterials with biocompatibility must be needed. In this chapter, we introduce collagen and hyaluronic acid (HA) as extracellular matrix as well as deal with the molecular mechanism as microenvironment, mechanistic effects, and gene expression. Application of collagen and HA have been reviewed in the area of orthopedics, orthopedics, ophthalmology, dermatology and plastic surgery. Finally, the ongoing and commercial products of collagen and HA for regenerative medicine have been introduced.