Evaluation of reverse transcription yield of RNA standards and forensic samples based on droplet digital PCR.

RNA analysis has shown great value in forensic science, such as body fluids and tissue identification, postmortem interval estimation, biological age prediction, etc. Currently, most RNA follow-up experiments involve reverse transcription (RT) procedures. It has been shown that the RT step is variable and has a greater impact on subsequent data analysis, especially for forensic trace samples. However, the pattern of variation between different RNA template inputs and complementary DNA (cDNA) yield is unclear. In this study, a series of 2-fold gradient dilutions of RNA standards (1 µg/µL - 0.24 ng/µL) and forensic samples (including blood samples, saliva samples, bloodstains, and saliva stains) were reverse-transcribed using EasyQuick RT MasterMix. The obtained cDNA was quantified by droplet digital PCR (ddPCR) to assess the RT yield of the ACTB gene. The results showed that the 125 ng RNA template had the highest RT yield in a 10 µL RT reaction system with the selected kit. For all stain samples, the RT yield improved as the amount of RNA template input increased since RNA quantities were below 125 ng. As many commercialized reverse transcription kits using different kinds of enzymes are available for forensic RNA research, we recommend that systematic experiments should be performed in advance to determine the amount of RNA input at the optimum RT yield when using any kit for reverse transcription experiments.

Leveraging Large Language Models for Improved Patient Access and Self-Management: Assessor-Blinded Comparison Between Expert- and AI-Generated Content.

BACKGROUND: While large language models (LLMs) such as ChatGPT and Google Bard have shown significant promise in various fields, their broader impact on enhancing patient health care access and quality, particularly in specialized domains such as oral health, requires comprehensive evaluation. OBJECTIVE: This study aims to assess the effectiveness of Google Bard, ChatGPT-3.5, and ChatGPT-4 in offering recommendations for common oral health issues, benchmarked against responses from human dental experts. METHODS: This comparative analysis used 40 questions derived from patient surveys on prevalent oral diseases, which were executed in a simulated clinical environment. Responses, obtained from both human experts and LLMs, were subject to a blinded evaluation process by experienced dentists and lay users, focusing on readability, appropriateness, harmlessness, comprehensiveness, intent capture, and helpfulness. Additionally, the stability of artificial intelligence responses was also assessed by submitting each question 3 times under consistent conditions. RESULTS: Google Bard excelled in readability but lagged in appropriateness when compared to human experts (mean 8.51, SD 0.37 vs mean 9.60, SD 0.33; P=.03). ChatGPT-3.5 and ChatGPT-4, however, performed comparably with human experts in terms of appropriateness (mean 8.96, SD 0.35 and mean 9.34, SD 0.47, respectively), with ChatGPT-4 demonstrating the highest stability and reliability. Furthermore, all 3 LLMs received superior harmlessness scores comparable to human experts, with lay users finding minimal differences in helpfulness and intent capture between the artificial intelligence models and human responses. CONCLUSIONS: LLMs, particularly ChatGPT-4, show potential in oral health care, providing patient-centric information for enhancing patient education and clinical care. The observed performance variations underscore the need for ongoing refinement and ethical considerations in health care settings. Future research focuses on developing strategies for the safe integration of LLMs in health care settings.

Impact of periodontitis on type 2 diabetes: a bioinformatic analysis.

BACKGROUND: Periodontitis is strongly associated with type 2 diabetes (T2D) that results in serious complications and mortality. However, the pathogenic role of periodontitis in the development of T2D and the underlain mechanism have not been fully elucidated. METHODS: A Mendelian randomization (MR) was performed to estimate the causality between two diseases. Bioinformatics tools, including gene ontology and pathway enrichment analyses, were employed to analyze the common differentially expressed genes (DEGs) in periodontitis and T2D. MR and colocalization analyses were then utilized to investigate the causal associations between potential pathogenic gene expression and the risk of T2D. Single cell-type expression analysis was further performed to detect the cellular localization of these genes. RESULTS: Genetically predicted periodontitis was associated with a higher risk of T2D (OR, 1.469; 95% CI, 1.117-1.930; P = 0.006) and insulin resistance (OR 1.034; 95%CI 1.001-1.068; P = 0.041). 79 common DEGs associated with periodontitis and T2D were then identified and demonstrated enrichment mainly in CXC receptor chemokine receptor binding and interleutin-17 signaling pathway. The integration of GWAS with the expression quantitative trait locis of these genes from the peripheral blood genetically prioritized 6 candidate genes, including 2 risk genes (RAP2A, MCUR1) and 4 protective genes (WNK1, NFIX, FOS, PANX1) in periodontitis-related T2D. Enriched in natural killer cells, RAP2A (OR 4.909; 95% CI 1.849-13.039; P = 0.001) demonstrated high risk influence on T2D, and exhibited strong genetic evidence of colocalization (coloc.abf-PPH4 = 0.632). CONCLUSIONS: This study used a multi-omics integration method to explore causality between periodontitis and T2D, and revealed molecular mechanisms using bioinformatics tools. Periodontitis was associated with a higher risk of T2D. MCUR1, RAP2A, FOS, PANX1, NFIX and WNK1 may play important roles in the pathogenesis of periodontitis-related T2D, shedding light on the development of potential drug targets.

Rapid and visual detection of specific bacteria for saliva and vaginal fluid identification with the lateral flow dipstick strategy.

Identification of body fluids is critical for crime scene reconstruction, and a source of investigation source of investigative leads. In recent years, microbial DNA analysis using sequencing and quantitative real-time polymerase chain reaction have been used to identify body fluids. However, these techniques are time-consuming, expensive, and require complex workflows. In this study, a new method for simultaneous detection of Streptococcus salivarius and Lactobacillus crispatus using polymerase chain reaction (PCR) in combination with a lateral flow dipstick (LFD) was developed to identify saliva and vaginal fluid in forensic samples. LFD results can be observed with the naked eye within 3 min with a sensitivity of 0.001 ng/µL DNA. The PCR-LFD assay was successfully used to detect S. salivarius and L. crispatus in saliva and vaginal fluid respectively, and showed negative results in blood, semen, nasal fluid, and skin. Moreover, saliva and vaginal fluid were detectable even at an extremely high mixing ratio of sample DNA (1:999). Saliva and vaginal fluid were identified in various mock forensic samples. These results indicate that saliva and vaginal fluid can be effectively detected by identifying S. salivarius and L. crispatus, respectively. Furthermore, we have shown that DNA samples used to identify saliva and vaginal fluid can also provide a complete short tandem repeat (STR) profile when used as source material for forensic STR profiling. In summary, our results suggest that PCR-LFD is a promising assay for rapid, simple, reliable, and efficient identification of body fluids.

Multiple/Two-Way Shape Memory Poly(urethane-urea-amide) Elastomers.

Multiple and two-way reversible shape memory polymers (M/2W-SMPs) are highly promising for many fields due to large deformation, lightweight, strong recovery stress, and fast response rates. Herein, a semi-crystalline block poly(urethane-urea-amide) elastomers (PUUAs) are prepared by the copolymerization of isocyanate-terminated polyurethane (OPU) and amino-terminated oligomeric polyamide-1212 (OPA). PUUAs, composed of OPA as stationary phase and PTMEG as reversible phase, exhibit excellent rigidity, flexibility, and resilience, and cPUUA-C7 -S25 exhibits the best tensile property with strength of 10.3 MPa and elongation at break of 360.2%. Besides, all the PUUAs possess two crystallization/melting temperatures and a glass transition temperature, which endow PUUAs with multiple and reversible two-way shape memory effect (M/2W-SME). Physically crosslinked PUUA-C0 -S25 exhibits excellent dual and triple shape memory, and micro chemically crosslinked cPUUA-C7 -S25 further shows quadruple shape memory behavior. Additionally, both PUUA-C0 -S25 and cPUUA-C7 -S25 have 2W-SME. Intriguingly, cPUUA-C7 -S25 can achieve a higher temperature (up to 165 °C) SME, which makes it suitable for more complex and changeable applications. Based on the advantages of M/2W-SME, a temperature-responsive application scenario where PUUAs can transform spontaneously among different shapes is designed. These unique M/2W-SME and high-temperature SME will enable the applications of high-temperature sensors, actuators, and aerospace equipment.

One-piece versus two-piece zirconia abutment supported single implant crown in the esthetic region: 3-Year results from a split-mouth randomized controlled clinical trial.

OBJECTIVES: To compare the clinical, radiographic, and immunological outcomes between one-piece versus two-piece zirconia abutments supported single implant crowns in the esthetic region. MATERIALS AND METHODS: The study followed a split-mouth, double-blind, and randomized controlled clinical design for a duration of 3 years. Twenty-two eligible patients with 44 implants were randomly assigned to two groups: Group 1 (one-piece zirconia abutment with zirconia base, n = 22) and Group 2 (two-piece zirconia abutment with titanium base, n = 22). The primary outcome was the technical complication rate. Additionally, survival rates, cytokines concentrations in peri-implant crevicular fluid (PICF), peri-implant conditions, marginal bone loss, and pink/white esthetics score (PES/WES) were assessed as secondary outcomes. RESULTS: Twelve of 22 patients attended the 1-year follow-up (due to the COVID pandemic), and 19 patients attended the 3-year examination. Two abutments in Group 1 were fractured after 10 and 12 months in function. Additionally, one screw loosening occurred in Group 1 at 1-year follow-up. The 3-year technical complication rate was significantly higher in Group 1 than that in Group 2 (15.79% vs. 0%, p < .001). The 3-year implant survival rate was 100% in both groups. The concentration of IFN-γ in PICF was significantly upregulated in Group 2 (p = .018). Furthermore, the IL-6 concentration was positively correlated with BOP% (p = .020). CONCLUSIONS: Two-piece zirconia abutments exhibited superior technical performance compared to one-piece designs during a 3-year follow-up in the anterior region. However, further long-term research is necessary to verify the immunological stability of two-piece zirconia abutments.

Effects of diabetes/hyperglycemia on peri-implant biomarkers and clinical and radiographic outcomes in patients with dental implant restorations: A systematic review and meta-analysis.

OBJECTIVE: The objective of this study is to systematically analyze the effects of diabetes mellitus/hyperglycemia (DM/HG) on peri-implant biomarkers and clinical and radiographic outcomes in patients undergoing implant treatment. MATERIALS AND METHODS: PubMed, Embase, and the Cochrane Central Register of Controlled Trials databases were searched up to March 2022. Studies reporting on peri-implant biomarkers, and clinical and radiographic outcomes in patients with different glycemic status levels (HbA1c) were included. Several analyses were conducted, such as meta-analysis, sensitivity analysis, dose-response meta-analysis (DRMA), and robust error meta-regression (REMR). RESULTS: A total of 10 studies and 634 participants were included for analysis, with a maximum follow-up of six years in function. The level of advanced glycation end products (AGEs) in peri-implant crevicular fluid (PICF) was significantly higher in the DM/HG group than in the healthy group (p < .01). Subgroup analyses showed that a set of negative regulators of bone metabolism (including IL-6, TNF-α, IL-8, and RANKL) were significantly higher in the DM/HG group (p = .01). Implant survival rate (100%) was not compromised in patients with an HbA1c level less than 10%. Outcomes of implant stability quotient (ISQ) values, bleeding on probing rate (BOP%), probing depth (PD), and marginal bone loss (MBL) in the DM/HG group were significantly worse than those in the healthy group (p = .04, <0.01, 0.01, <0.01, respectively). DRMA results showed AGE accumulation in PICF, PD, and MBL worsened in a dose-response dependent manner with elevated HbA1c levels (<6%, 6-8%, >8%; p = .04, 0.02, <0.01, respectively). CONCLUSIONS: DM/HG can upregulate negative regulators of bone metabolism and compromise peri-implant health. In addition, there are dose-response relationships between HbA1c levels and AGE accumulation in PICF, PD, and MBL.

Accuracy and primary stability of tapered or straight implants placed into fresh extraction socket using dynamic navigation: a randomized controlled clinical trial.

OBJECTIVES: To compare the accuracy and primary stability of tapered and straight implants undergoing immediate implant placement with dynamic navigation. MATERIALS AND METHODS: Patients with compromised anterior teeth in maxilla were recruited and allocated randomly into (1) tapered implant group (TI group) and (2) straight implant group (SI group). Implants were inserted into fresh sockets with dynamic navigation. Three-dimensional platform deviation, apex deviation, angular deviation, insertion torque value (ITV) and implant stability quotient (ISQ) were recorded. RESULTS: Twenty patients with 20 implants were included. The overall platform, apex, and angular deviation were 0.87 ± 0.35 mm, 0.81 ± 0.34 mm, and 2.40 ± 1.31°, respectively. The accuracy was 0.86 ± 0.26 mm, 0.76 ± 0.33 mm, and 2.49 ± 1.54° for TI, and 0.89 ± 0.44 mm, 0.88 ± 0.36 mm, and 2.31 ± 1.01° for SI, with no significant difference (p = 0.85, 0.45, 0.76). Sagittal root position classification (SRP) class I may obtain greater error in numerical values in straight implants (0.97 ± 0.47 mm vs. 0.6 ± 0.16 mm, 0.92 ± 0.36 mm vs. 0.73 ± 0.36 mm, 2.48 ± 1.19° vs. 1.71 ± 0.14°). The overall ISQ was 60.74. ISQ was 60.48 for TI and 60.96 for SI, with no significant difference. Acceptable ITV (> 15 Ncm) was achieved in most of the included patients (SI 7/10, TI 9/10). CONCLUSIONS: High accuracy and primary stability of immediate implant placement could be achieved both in tapered and straight implants with dynamic navigation systems. CLINICAL RELEVANCE: Tapered and straight implants did not reach a consensus on which was better in immediate implant regarding to accuracy and primary stability. Our study demonstrated implant macrodesign did not affect accuracy and primary stability in immediate implant using dynamic navigation.

Remodeling immune microenvironment in periodontitis using resveratrol liposomes as an antibiotic-free therapeutic strategy.

BACKGROUND: Periodontitis is a complicated inflammatory disease that damages the tooth-supporting tissues, with limited pharmacotherapy available. Macrophage-targeting therapy is promising for inflammatory diseases. Resveratrol (RSV), a nonflavonoid polyphenol, is known for its anti-inflammatory and immunomodulatory effects. However, its medical application is limited by its poor stability and water-solubility, as well as its low bioavailability. RESULT: A therapeutic resveratrol-loaded liposomal system (Lipo-RSV) was developed to treat periodontitis. The physical properties of Lipo-RSV and its ability to regulate macrophages were investigated. The results showed that Lipo-RSV had good biocompatibility and could re-educate the inflammatory macrophages from M1- to M2-like phenotype through activating p-STAT3 and downregulating p-STAT1. Besides, the Lipo-RSV could scavenge ROS and inhibit the NF-κB signal and inflammasomes, thereby reducing the pro-inflammatory cytokines IL-1ß, IL-6, and TNF-α. CONCLUSION: These results revealed that Lipo-RSV could be a potential therapeutic system for the antibiotic-free treatment for periodontal diseases.

Novel application of bergapten and quercetin with anti-bacterial, osteogenesis-potentiating, and anti-inflammation tri-effects.

The bacteria-mediated inflammatory conditions adversely affect the osseointegration process of endosseous implants, which can even lead to implant malfunction or failure. Local drug delivery has been designed to exert anti-inflammatory and antibacterial activities, but whether this strategy has an effect on the compromised osseointegration under inflammation has rarely been studied. The present study focused on the osteoinductive efficacy of two known phytoestrogens [bergapten (BP) and quercetin (QE)] on implant sites under multiple bacteria-infected conditions in situ. Furthermore, the gene expression profiles of rat bone mesenchymal stem cells (rBMSCs) treated with BP and QE in the presence of Porphyromonas gingivalis-derived lipopolysaccharide were identified. The results showed that both drugs, especially QE, had significant potentiating effects on promoting osteogenic differentiation of rBMSCs, resisting multiple pathogens, and reducing inflammatory activity. Meanwhile, RNA sequencing analysis highlighted the enriched gene ontology terms and the differentially expressed genes (Vps25, Il1r2, Csf3, Efemp1, and Ccl20) that might play essential roles in regulating the above tri-effects, which provided the basis for the drug delivery system to be used as a novel therapeutic strategy for integrating peri-implant health. Overall, our study confirmed that QE appeared to outperform BP in osteogenesis and bacterial killing but not in anti-inflammation. Moreover, both drugs possess favorable tri-effects and can serve as the pivotal agents for the drug delivery system to boost osseointegration at inflammatory implant sites.

A novel semiautomatic segmentation protocol to evaluate guided bone regeneration outcomes: A pilot randomized, controlled clinical trial.

OBJECTIVES: The aims of this study were to (a) present a novel morphological contour interpolation (MCI) algorithm based method to evaluate grafted bone alterations following guided bone regeneration (GBR), (b) compare clinical and radiological outcomes of GBR with two different collagen membranes. MATERIALS AND METHODS: The data were retrieved from an ongoing randomized controlled trial. Patients were randomly allocated into two groups: (a) control group (CG): Bio-Gide (b) test group (TG): bovine dermis-derived collagen membrane. Cone beam computed tomography examinations were performed 1 week (T0) and 6 months after surgery (T1). PES/WES at T1, grafted bone volume and density changes from T0 to T1 were recorded. RESULTS: Thirty-six patients (16/20 in test/control group, respectively) were enrolled in the present study. Excellent inter-observer reliability (ICC ≥ 0.97) was revealed for repeated measurements using this method. Significant volumetric reduction of grafted bone were found in both groups (test group: from 0.60 to 0.39 cm3 , p < 0.01; control group: from 0.54 to 0.31 cm3 , p < 0.01). Mean bone density (gray-scale values) significantly increased from 305.12 to 456.69 in CG (p < 0.01). In TG, it slightly increased from 304.75 to 393.27 (p = 0.25). The mean PES/WES values were 13.84 (6.62/7.22) and 13.90 (6.70/7.20) for TG and CG, respectively. As for inter-group comparison, no significant differences of grafted bone volume change, density change and PES/WES were found between two groups. CONCLUSION: Within the limitations of this study, the novel MCI-based method is a reproducible tool to segment and visualize changes of grafted bone in 3D. Furthermore, both collagen membranes could be used as a barrier membrane for GBR in humans.

Hardware complications and failure of three-unit zirconia-based and porcelain-fused-metal implant-supported fixed dental prostheses: a retrospective cohort study with up to 8 years.

OBJECTIVE: The aim of the present study was to assess the hardware complications and survival of three-unit implant-supported zirconia-based fixed dental prostheses (IZ) and implant-supported porcelain-fused-metal fixed dental prostheses (IP). MATERIAL AND METHODS: The study is a retrospective cohort study with up to 8 years (mean 4.8 years) follow-up. Patients with conventional three-unit implant-supported fixed dental prostheses (without cantilever) in posterior area were reviewed. Hardware complications and survival rate were evaluated. Chi-Square test was used to test the difference between IZ and IP groups. RESULTS: Two hundred and thirty-seven patents (IZ : 112, IP : 125) with 279 three-unit restorations (IZ : 127, IP : 152) participated in the study. The overall survival rate was 95.3% in IZ group and 94.7% in IP group at implant-level and 94.6% in IZ group and 94.4% in IP group at subject-level. Veneer chipping was the most frequently seen complication (20.1%). Significant higher minor veneer chipping rate (Grade 1) was found in IZ group (P = 0.04). No significant difference of veneer chipping rate (Grade 2 and Grade 3) was found between the two groups. The overall hardware complication rates of IZ were significantly higher than IP (33.07% and 18.42%, P = 0.01). CONCLUSION: High survival rate of zirconia-base and Porcelain-fused-to-metal (PFM) restorations can be achieved with up to 8 years follow-up. The zirconia-based restorations need more polishing procedures, maintenance, and professional care than PFM restorations. The number of implants supporting a fixed dental prosthesis did not influence the implant survival and hardware complications. Well-designed studies with high evidence level are still needed to further explore the hardware complications and clinical survival of IZ and IP.

Dioscin Alleviates Periodontitis by Inhibiting NLRP3 Inflammasome Activation via Regulation of K+ Homeostasis and Mitochondrial Function.

Gingival inflammation and alveolar bone loss are characteristic manifestations of periodontitis. Interleukin (IL)-1ß, the maturation of which is mainly regulated by NOD-like receptor protein (NLRP) 3 inflammasome, not only amplifies the inflammatory response but also triggers osteoclastogenesis, thereby accelerating the progression of periodontitis. Dioscin, a natural steroid saponin, has been shown to inhibit NLRP3 inflammasome. Nevertheless, research on the effectiveness of Dioscin for the management of periodontitis remains scarce. In this study, Dioscin was found to dramatically reduce the integral components of NLRP3 inflammasome, ultimately limiting IL-1ß secretion. Notably, the inhibitory impact of Dioscin on NLRP3 inflammasome might be exerted by curbing the generation of mitochondrial (mt) reactive oxygen species (ROS) and oxidized (ox) mtDNA, which were mediated by inhibition of K+ efflux. Furthermore, Dioscin effectively alleviated periodontitis in mice. Overall, the results established that Dioscin could alleviate periodontitis by inhibiting NLRP3 inflammasome via modulation of the K+ efflux-mtROS-ox-mtDNA pathway, holding the potential to treat periodontitis and other NLRP3-driven inflammatory diseases.

Biocompatible and Biodegradable 3D Graphene/Collagen Fiber Hybrids for High-Performance Conductive Networks and Sensors.

Polymer-based flexible conductive materials are crucial for wearable electronics, electronic skin, and other smart materials. However, their development and commercial applications have been hampered by the lack of strain tolerance in the conductive network, poor bonding with polymers, discomfort during wear, and a lack of biocompatibility. This study utilized oil-tanned leather with a natural network structure, high toughness, and high tensile deformation recovery as a structural template. A graphene (Gr) conductive network was then constructed on the collagen network of the leather, with coordination cross-linking between Gr and collagen fibers through aluminum ions (Al3+). A new flexible conductive material (Al-GL) was then constructed. Molecular dynamics simulations and experimental validation revealed the existence of physical adsorption, hydrogen bonding adsorption, and ligand bonding between Al3+, Gr, and collagen fibers. Although we established that the binding sites between Al3+ and collagen fibers were primarily on carboxyl groups (-COOH), the mechanism of chemical bonding between Gr and collagen fibers remains unclear. The Al-GL composite exhibited a high shrinkage temperature (67.4 °C) and low electrical resistance (16.1 kΩ·sq-1), as well as good softness (9.33 mN), biocompatibility, biodegradability (<60 h), and air and moisture permeability. Furthermore, the incorporation of Al3+ resulted in a heightened Gr binding strength on Al-GL, and the resistance remained comparable following 1 h of water washing. The Al-GL sensor prepared by WPU encapsulation not only demonstrated highly sensitive responses to diverse motion signals of the human body but also retained a certain degree of response to external mechanical effects underwater. Additionally, the Al-GL-based triboelectric nanogenerator (Al-GL TENG) exhibited distinct response signals to different materials. The Al-GL prepared by the one-pot method proposed in this study offers a novel approach to combining functional nanofillers and substrate materials, providing a theoretical foundation for collagen fiber-based flexible conductive materials.

Strategies for Treating Traumatic Neuromas with Tissue-Engineered Materials.

Neuroma, a pathological response to peripheral nerve injury, refers to the abnormal growth of nerve tissue characterized by disorganized axonal proliferation. Commonly occurring after nerve injuries, surgeries, or amputations, this condition leads to the formation of painful nodular structures. Traditional treatment options include surgical excision and pharmacological management, aiming to alleviate symptoms. However, these approaches often offer temporary relief without addressing the underlying regenerative challenges, necessitating the exploration of advanced strategies such as tissue-engineered materials for more comprehensive and effective solutions. In this study, we discussed the etiology, molecular mechanisms, and histological morphology of traumatic neuromas after peripheral nerve injury. Subsequently, we summarized and analyzed current nonsurgical and surgical treatment options, along with their advantages and disadvantages. Additionally, we emphasized recent advancements in treating traumatic neuromas with tissue-engineered material strategies. By integrating biomaterials, growth factors, cell-based approaches, and electrical stimulation, tissue engineering offers a comprehensive solution surpassing mere symptomatic relief, striving for the structural and functional restoration of damaged nerves. In conclusion, the utilization of tissue-engineered materials has the potential to significantly reduce the risk of neuroma recurrence after surgical treatment.

Body fluids should be identified before estimating the time since deposition (TsD) in microbiome-based stain analyses for forensics.

Identification and the time since deposition (TsD) estimation of body fluid stains from a crime scene could provide valuable information for solving the cases and are always difficult for forensics. Microbial characteristics were considered as a promising biomarker to address the issues. However, changes in the microbiota may damage the specific characteristics of body fluids. Correspondingly, incorrect body fluid identification may result in inaccurate TsD estimation. The mutual influence is not well understood and limited the codetection. In the current study, saliva, semen, vaginal secretion, and menstrual blood samples were exposed to indoor conditions and collected at eight time points (from fresh to 30 days). High-throughput sequencing based on the 16S rRNA gene was performed to characterize the microbial communities. The results showed that a longer TsD could decrease the discrimination of different body fluid stains. However, the accuracies of identification still reached a quite high value even without knowing the TsD. Correspondingly, the mean absolute error (MAE) of TsD estimation significantly increased without distinguishing the types of body fluids. The predictive TsD of menstrual blood reached a quite low MAE (1.54 ± 0.39 d). In comparison, those of saliva (6.57 ± 1.17 d), semen (6.48 ± 1.33 d), and vaginal secretion (5.35 ± 1.11 d) needed to be further improved. The great effect of individual differences on these stains limited the TsD estimation accuracy. Overall, microbial characteristics allow for codetection of body fluid identification and TsD estimation, and body fluids should be identified before estimating TsD in microbiome-based stain analyses.IMPORTANCEEmerged evidences suggest microbial characteristics could be considered a promising tool for identification and time since deposition (TsD) estimation of body fluid stains. However, the two issues should be studied together due to a potential mutual influence. The current study provides the first evidence to understand the mutual influence and determines an optimal process for codetection of identification and TsD estimation for unknown stains for forensics. In addition, we involved aged stains into our study for identification of body fluid stains, rather than only using fresh stains like previous studies. This increased the predictive accuracy. We have preliminary verified that individual differences in microbiotas limited the predictive accuracy of TsD estimation for saliva, semen, and vaginal secretion. Microbial characteristics could provide an accurate TsD estimation for menstrual blood. Our study benefits the comprehensive understanding of microbiome-based stain analyses as an essential addition to previous studies.

Clinical, radiographic, and esthetic evaluation of immediate implant placement with buccal bone dehiscence in the anterior maxilla: A 1-year prospective case series.

OBJECTIVES: To evaluate the clinical, radiographic, and esthetic outcomes of immediate implant placement with buccal bone dehiscence in the anterior maxilla. METHODS: In this case series, implants were inserted immediately after tooth extraction in sockets with buccal bone dehiscence. Guided bone regeneration (GBR) with a papilla preservation flap and simultaneous connective tissue grafting (CTG) was used. The following outcome variables were measured: mid-facial mucosal recession, probing depth, bleeding on probing, Pink Esthetic Score (PES), marginal bone loss, and thickness of buccal bone plate (TBP). RESULTS: 12 patients were recruited. Stable mid-facial mucosal level (-0.03 ± 0.17 mm) and excellent soft-tissue esthetic outcomes (PES, 9.17 ± 0.72) were achieved at 1 year. The TBP at platform level was 2.01 ± 0.31 mm at 1-year follow up with a resorption rate of 28.90% ± 15.14%. CONCLUSIONS: Immediate implant placement using GBR performed with a papilla preservation approach and simultaneous CTG is a feasible treatment procedure in compromised extraction sockets in the anterior region. Favorable esthetic outcomes and buccal bone thickness were obtained. Further studies were needed to evaluate the long-term tissue alteration.

Fabrication and Characterization of Collagen/PVA Dual-Layer Membranes for Periodontal Bone Regeneration.

Guided tissue regeneration (GTR) is a promising treatment for periodontal tissue defects, which generally uses a membrane to build a mechanical barrier from the gingival epithelium and hold space for the periodontal regeneration especially the tooth-supporting bone. However, existing membranes possess insufficient mechanical properties and limited bioactivity for periodontal bone regenerate. Herein, fish collagen and polyvinyl alcohol (Col/PVA) dual-layer membrane were developed via a combined freezing/thawing and layer coating method. This dual-layer membrane had a clear but contact boundary line between collagen and PVA layers, which were both hydrophilic. The dual membrane had an elongation at break of 193 ± 27% and would undergo an in vitro degradation duration of more than 17 days. Further cell experiments showed that compared with the PVA layer, the collagen layer not only presented good cytocompatibility with rat bone marrow-derived mesenchymal stem cells (BMSCs), but also promoted the osteogenic genes (RUNX2, ALP, OCN, and COL1) and protein (ALP) expression of BMSCs. Hence, the currently developed dual-layer membranes could be used as a stable barrier with a stable degradation rate and selectively favor the bone tissue to repopulate the periodontal defect. The membranes could meet the challenges encountered by GTR for superior defect repair, demonstrating great potential in clinical applications.

Case Report: A Case Report and Literature Review of 3p Deletion Syndrome.

Objective: The aim of the present study is to explore the clinical and genetic characteristics of 3p deletion syndrome to improve clinicians' understanding of the disease. Methods: The clinical manifestations, process of diagnosis and treatment, and genetic characteristics of an individual case of 3p deletion syndrome were analyzed. CNKI, Wanfang Data, and the Biomedical Literature Database (PubMed) were searched. The search time limit, using "3p deletion syndrome" and "BRPF1" as keywords, was from the creation of the database up to June 2020. Related data were reviewed. Results: The proband was a male child with general developmental and intellectual disabilities, special facial features and congenital heart disease. The child was the parents' first pregnancy and first born. Gene microarray analysis showed a 10.095 Mb deletion in the 3p26.3-p25.3 region, resulting in a heterozygous mutation of the BRPF1 gene; thus, the patient was diagnosed with 3p deletion syndrome. At the time of diagnosis, the child was 1 year of age and was responding to comprehensive rehabilitation training. A total of 29 well-documented cases were found in the literature, of which 19 cases had an onset within 1 year of birth, and mainly manifested with mental and motor development disabilities and abnormal facial features, with different gene deletions, depending on the size and location of the 3p deletion. Conclusion: The genetic test results of the child in this study indicated a heterozygous deletion of the BRPF1 gene on the short arm of chromosome 3, which was a unique feature of this study, since it was rarely mentioned in other reports of 3p deletion syndrome. The clinical phenotype of this syndrome is complex as it can include intellectual and motor development backwardness, low muscle tone, certain abnormal facial features (low hairline, bilateral ptosis, widely spaced eyes, a forward nose, left ear auricle deformity, a high-arched palate, a small jaw), and the deformation of systems such as the gastrointestinal tract and the urinary tract malformation or symptoms of epilepsy. As clinical manifestations can be relatively mild, the syndrome is easy to miss or misdiagnose. Clinical workers need to be aware of this disease when they find that children have special features, such as stunted growth, low muscle tone or ptosis, and it needs to be diagnosed through genetic testing. Most children are able to develop certain social skills after rehabilitation treatment.

From Unimolecular Template to Silver Nanocrystal Clusters: An Effective Strategy to Balance Antibacterial Activity and Cytotoxicity.

Silver nanomaterials have attracted a great deal of interest due to their broad-spectrum antimicrobial activity. However, it is still challenging to balance the high antibacterial efficiency with low damage to biological cells of silver nanostructures, especially when the diameter decreases to less than 10 nm. Here, we developed a new type of Ag nanohybrid material via a unimolecular micelle template method, which presents amazing antibacterial activities and almost noncytotoxicity. First, water-soluble multiarm star-shaped brushlike copolymer α-CD-g-[(PEO40-g-PAA50)-b-PEO5]18 was precisely synthesized and its micelle behavior in different solvents was revealed. Then, nanocrystal clusters assembled by Ag grains (Ag@Template NCs) were prepared through an in situ redox route using the unimolecular micelle of α-CD-g-[(PEO40-g-PAA50)-b-PEO5]18 as the soft template, AgNO3 as a precursor, and tetrabutylammonium borohydride (TBAB) as the reducing agent. The overall size of the achieved Ag@Template NCs is controlled by the template structure at around 40 nm (Dh in DMF), and the size of the Ag grain can be easily regulated from ∼1 to ∼5 nm by adjusting the feeding ratio of AgNO3/acrylic acid (AA) units in the template from 1:10 to 1:1. Benefitting from the structural design of the template, all Ag@Template NCs prepared here exhibit excellent dispersibility and chemical stability in different aqueous environments (neutral, pH = 5.5, and 0.9% NaCl physiological saline solution), which play a crucial role in the long-term storage and potential application in a complex physiological environment. The antibacterial and cytotoxicity tests indicate that Ag@Template NCs display much better performance than Ag nanoparticles (Ag NPs), which have a comparable overall size of ∼25 nm. The inhibitory capability of Ag@Template NCs to bacteria strongly depends on the grain size. Specifically, the Ag@Template-1 NC assembled by the smallest grains (1.6 ± 0.3 nm) presents the best antibacterial activity. For E. coli (-), the MIC value is as low as 5 µg/mL (0.36 µg/mL of Ag), while for S. aureus (+), the value is around 10 µg/mL (0.72 µg/mL of Ag). The survival rate of L02 cells and lactate dehydrogenase assay together illustrate the low cytotoxicity possessed by the prepared Ag@Template NCs. Therefore, the proposed Ag@Template NC structure successfully resolves the high reactivity, instability, and fast oxidation issues of the ultrasmall Ag nanoparticles, and integrates high antibacterial efficiency and nontoxicity to biological cells into one platform, which implies its broad potential application in biomedicine.