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Cheng-Shyuan Rau1 *, Pao-Jen Kuo2 *, Hui-Ping Lin,2 Chia-Jung Wu,2 Yi-Chan Wu,2 Peng-Chen Chien,2 Ting-Min Hsieh,3 Hang-Tsung Liu,3 Chun-Ying Huang,3 Ching-Hua Hsieh2 1Department of Neurosurgery, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, Kaohsiung, Taiwan; 2Department of Plastic Surgery, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, Kaohsiung, Taiwan; 3Department of Trauma Surgery, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, Kaohsiung, Taiwan*These authors contributed equally to this workCorrespondence: Ching-Hua Hsieh, Department of Plastic Surgery, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, No. 123, Ta-Pei Road, Niao-Song District, Kaohsiung City, 833, Taiwan, Tel +886-7-7327476, Fax +886-7-3450431, Email m93chinghua@gmail.comPurpose: Following major trauma, genes involved in adaptive immunity are downregulated, which accompanies the upregulation of genes involved in systemic inflammatory responses. This study investigated microRNA (miRNA)-mRNA interactome dysregulation in circulating T cells of patients with major trauma.Patients and Methods: This study included adult trauma patients who had an injury severity score ≥ 16 and required ventilator support for more than 48 h in the intensive care unit. Next-generation sequencing was used to profile the miRNAs and mRNAs expressed in CD3+ T cells isolated from patient blood samples collected during the injury and recovery stages.Results: In the 26 studied patients, 9 miRNAs (hsa-miR-16-2-3p, hsa-miR-16-5p, hsa-miR-185-5p, hsa-miR-192-5p, hsa-miR-197-3p, hsa-miR-23a-3p, hsa-miR-26b-5p, hsa-miR-223-3p, and hsa-miR-485-5p) were significantly upregulated, while 58 mRNAs were significantly downregulated in T cells following major trauma. A network consisting of 8 miRNAs and 22 mRNAs interactions was revealed by miRWalk, with three miRNAs (hsa-miR-185-5p, hsa-miR-197-3p, and hsa-miR-485-5p) acting as hub genes that regulate the network. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis suggested that “chemokine signaling pathway” was the predominant pathway.Conclusion: The study revealed a miRNA-mRNA interactome consisting of 8 miRNAs and 22 mRNAs that are predominantly involved in chemokine signaling in circulating T cells of patients following major trauma.Keywords: miRNA, next-generation sequencing, T cells, adaptive immunity, major trauma, critical illness

Shao-Chun Wu,1,* Cheng-Shyuan Rau,2,* Pao-Jen Kuo,3 Fu-Yuan Shih,2 Hui-Ping Lin,3 Yi-Chan Wu,3 Ting-Min Hsieh,4 Hang-Tsung Liu,4 Ching-Hua Hsieh3 1Department of Anesthesiology, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, Kaohsiung, Taiwan; 2Department of Neurosurgery, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, Kaohsiung, Taiwan; 3Department of Plastic Surgery, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, Kaohsiung, Taiwan; 4Department of Trauma Surgery, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, Kaohsiung, Taiwan*These authors contributed equally to this workCorrespondence: Ching-Hua HsiehDepartment of Plastic Surgery, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, No. 123, Ta-Pei Road, Niao-Song District, Kaohsiung, 833, TaiwanTel +886-7-7327476Fax +886-7-3450431Email m93chinghua@gmail.comPurpose: Severe trauma may lead to the systemic release of inflammatory mediators into the circulation with profound acute-phase responses; however, the understanding of the expression of these mediators remains limited. This study aimed to characterize the alterations in the expression of circulating acute-phase proteins, cytokines, and checkpoint proteins in patients with severe trauma injuries.Patients and Methods: The study population included trauma patients in the intensive care unit (ICU) with an injury severity score equal to or greater than 16 and who had used a ventilator for 48 hours. A total of 12 female and 28 male patients were recruited for the study; six patients died and 34 survived. Blood samples collected at acute stages were compared with those drawn at the subacute stage, the time when the patients were discharged from the ICU, or before the discharge of the patients from the hospital.Results: The study identified that the expression of acute-phase proteins, such as alpha-1-acid glycoprotein and C-reactive protein, and cytokines, including granulocyte colony-stimulating factor, interleukin-6, and interleukin-1 receptor antagonist, was elevated in the circulation after severe trauma. In contrast, the levels of acute-phase proteins, such as alpha-2-macroglobulin, serum amyloid P, and von Willebrand factor, and cytokines, including interleukin-4 and interferon gamma-induced protein 10, were reduced. However, there were no significant differences in the expression of checkpoint proteins in the circulation.Conclusion: The dysregulated proteins identified in this study may serve as potential therapeutic targets or biomarkers for treating patients with severe trauma. However, the related biological functions of these dysregulated factors require further investigation to validate their functions.Keywords: circulation, inflammatory reaction, biomarkers, severe trauma, critical illness

The purpose of this technical report is to describe a modified tunnel surgical approach and connective tissue graft (CTG) stabilization technique for the treatment of gingival recessions with interproximal clinical attachment loss (ICAL). The partial-full-thickness (PFT) tunnel technique utilizes multiple vestibular incisions to facilitate creation of a split-mucoperiosteal tunnel that enhances tissue passivity and allows for coronal advancement of soft tissue with minimal tension. The supra-crestal sling (SCS) suture engages only the CTG, independent of the overlying tissue and stabilizes the CTG around the buccal and proximal root surfaces. The treatment approach of PFT tunnel preparation and CTG stabilization with the SCS suture was designed to optimize blood supply and maximize wound stability, resulting in complete root coverage with satisfactory clinical outcomes in RT2 and RT3 gingival recession. It is suggested that the PFT tunnel preparation with graft stabilization via the SCS suture has the potential to treat recessions with anatomical limitations associated with ICAL.

Background: The leakage of saliva through the deep neck region from a pharyngocutaneous fistula could cause devastating complications, including vascular ruptures leading to mortality. While a partial pharyngoesophageal defect is created after total laryngectomy, a patch pattern of hypopharyngeal reconstruction is required, for which a fasciocutaneous free flap is usually applied. If radiotherapy fails to cure pharyngeal cancer, salvage total laryngectomy (STL) is needed. However, postradiation tissues tend not to heal well, and the incidence of pharyngocutaneous fistula therefore increases. We proposed an edge-epithelialization method to address this problem and conducted a retrospective study for comparison., Methods: The inclusion criteria were patients with head and neck cancer who underwent total laryngectomy that immediately required patch free flap reconstruction at a single medical center (January 2012-December 2021). Receipt of presurgical radiotherapy, hospitalization duration, and the presence of postoperative complications were recorded., Results: The included patients were separated into two groups: Group A (edge de-epithelialization not adopted) (n = 79) and Group B (edge de-epithelialization adopted) (n = 51). Forty-four and twenty-two patients in Groups A and Group B, respectively, received preoperative radiotherapies and simultaneous STL and fasciocutaneous free flap reconstructions. The incidence of pharyngocutaneous fistula was significantly lower in Group B (p = 0.0145). This phenomenon was the same for patients who underwent preoperative radiotherapy only (p = 0.0470) but not for patients who did not receive preoperative radiotherapy (p = 0.2363)., Conclusions: Edge de-epithelialization is an effective method for reducing pharyngocutaneous fistula formation in patch free flap reconstructions after STLs., (© 2024. Society of Surgical Oncology.)

Background: The stress index (SI), defined as the serum glucose to potassium ratio, has emerged as a potential prognostic indicator in some patient populations. This study aims to evaluate the predictive value of SI on the trauma patients sustained by all trauma causes., Methods: A retrospective analysis was conducted on 20,040 adult trauma patients admitted to a single trauma center from January 1, 2009, to December 31, 2022. The SI was calculated according to the serum levels of glucose (mg/dL) and potassium (mEq/L) upon patients' arrival to emergency room. The enrolled patients were stratified into two groups based on an optimal SI cutoff value determined by receiver operating characteristic (ROC) curve analysis. The association between SI and in-hospital mortality, as well as other clinical outcomes, was assessed using multivariate logistic regression, adjusting for potential confounders., Results: The mortality patients had a significantly higher SI (59.7 ± 30.6 vs. 39.5 ± 17.5, p < 0.001) than those who survived. The SI was identified as a significant independent predictor of mortality (odds ratio [OR] 4.65, 95 % confidence interval [CI]: 2.61-8.27, p < 0.001) in the multivariate analysis. In addition, patients in the high SI group (≥42.7) demonstrated significantly worse outcomes, including higher in-hospital mortality (7.5 % vs. 1.4 %, p < 0.001), longer hospital stays compared to the low SI group (<42.7)., Conclusion: The SI serves as a simple and valuable prognostic tool in risk stratification of the trauma patients., Competing Interests: The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Ching-Hua Hsieh reports a relationship with 10.13039/100012553Chang Gung Memorial Hospital Kaohsiung Branch that includes: funding grants. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors.)

Natural products have attracted great interest in the development of tissue engineering. Recent studies have demonstrated that unsaturated fatty acids found in natural plant seed oil may exhibit positive osteogenic effects; however, few in vivo studies have focused on the use of plant seed oil for bone regeneration. The aim of this study is to investigate the effects of seed oil found in Sapindus mukorossi ( S. mukorossi ) on the osteogenic differentiation of mesenchymal stem cells and bone growth in artificial bone defects in vivo. In this study, Wharton-jelly-derived mesenchymal stem cells (WJMSCs) were co-cultured with S. mukorossi seed oil. Cellular osteogenic capacity was assessed using Alizarin Red S staining. Real-time PCR was carried out to evaluate ALP and OCN gene expression. The potential of S. mukorossi seed oil to enhance bone growth was assessed using an animal model. Four 6 mm circular defects were prepared at the parietal bone of New Zealand white rabbits. The defects were filled with hydrogel and hydrogel- S. mukorossi seed oil, respectively. Quantitative analysis of micro-computed tomography (Micro-CT) and histological images was conducted to compare differences in osteogenesis between oil-treated and untreated samples. Although our results showed no significant differences in viability between WJMSCs treated with and without S. mukorossi seed oil, under osteogenic conditions, S. mukorossi seed oil facilitated an increase in mineralized nodule secretion and upregulated the expression of ALP and OCN genes in the cells ( p < 0.05). In the animal study, both micro-CT and histological evaluations revealed that new bone formation in artificial bone defects treated with S. mukorossi seed oil were nearly doubled compared to control defects ( p < 0.05) after 4 weeks of healing. Based on these findings, it is reasonable to suggest that S. mukorossi seed oil holds promise as a potential candidate for enhancing bone healing efficiency in bone tissue engineering.

Background: Ferroptosis is an iron-driven cell-death mechanism that plays a central role in various diseases. Recent studies have suggested that baicalein inhibits ferroptosis, making it a promising therapeutic candidate. Materials and Methods: Fibroblast cultures were treated with different agents to determine the effects of baicalein on ferroptosis. Ferroptosis-related gene expression, lipid peroxidation, and post-treatment cellular structural changes were measured using real-time quantitative polymerase chain reaction, C11-BODIPY dye, and transmission electron microscopy, respectively. Results: Baicalein significantly inhibited rat sarcoma virus selective lethal 3-induced ferroptosis in fibroblasts. Moreover, in baicalein-treated groups, reduced ferroptosis-related gene expression, decreased lipid peroxidation, and maintained cell structure was observed when compared with those of the controls. Discussion: The ability of baicalein to counteract RSL3-induced ferroptosis underscores its potential protective effects, especially in diseases characterized by oxidative stress and iron overload in fibroblasts. Conclusion: Baicalein may serve as a potent therapeutic agent against conditions in which ferroptosis is harmful. The compound's efficacy in halting RSL3-triggered ferroptosis in fibroblasts paves the way for further in vivo experiments and clinical trials., Competing Interests: Competing Interests: The authors have declared that no competing interest exists., (© The author(s).)

The easy albumin-bilirubin (EZ-ALBI) score is derived using the following equation: total bilirubin (mg/dL) - 9 × albumin (g/dL). This study aimed to determine whether the EZ-ALBI score predicted mortality risk in adult trauma patients in an intensive care unit (ICU). Data from a hospital's trauma database were retrospectively evaluated for 1083 adult trauma ICU patients (139 deaths and 944 survivors) between 1 January 2016 and 31 December 2021. Patients were classified based on the ideal EZ-ALBI cut-off of -26.5, which was determined via receiver operating characteristic curve analysis. The deceased patients' EZ-ALBI scores were higher than those of the surviving patients (-26.8 ± 6.5 vs. -30.3 ± 5.9, p = 0.001). Multivariate logistic analysis revealed that, in addition to age, the presence of end-stage renal disease, Glasgow Coma Scale scores, and injury severity scores, the EZ-ALBI score is an independent risk factor for mortality (odds ratio (OR), 1.10; 95% confidence interval (CI): 1.06-1.14; p = 0.001)). Compared with patients with EZ-ALBI scores < -26.5, those with scores ≥ -26.5 had a 2.1-fold higher adjusted mortality rate (adjusted OR, 2.14; 95% CI: 1.43-3.19, p = 0.001). In conclusion, the EZ-ALBI score is a substantial and independent predictor of mortality and can be screened to stratify mortality risk in adult trauma ICU patients.

Purpose: This study aimed to investigate the relative frequency of congenital upper-limb anomalies (CULAs) in southern Taiwan using the 2020-updated the Oberg, Manske, and Tonkin (OMT) classification system and evaluate the practicality of the new classification system., Methods: We retrospectively reviewed patients with CULAs from 1987 to 2021 at a referral center in southern Taiwan. All patients were analyzed based on medical records, photographs, and radiographs, and the anomalies were classified according to the 2020 OMT classification system., Results: A total of 1,188 patients with 1,335 CULAs were retrospectively reviewed. The results demonstrated that the most common type of CULA was malformations (1,092 cases), followed by dysplasias (144 cases), syndromes (51 cases), and deformations (48 cases). Among the malformations, radial polydactyly was the most common anomaly (732 cases), followed by simple syndactyly (66 cases). Among the dysplasias, camptodactyly was the most common anomaly (52 cases), followed by thumb-in-palm deformity (45 cases) and vascular tumors (17 cases). In the deformations, constriction ring sequence accounted for all cases. Poland syndrome (21 cases) occurred most often in the category of syndromes., Conclusions: The results of this study show that radial polydactyly (732 cases, 55%) is the most common CULA in southern Taiwan, followed by simple syndactyly (66 cases, 5%) and then camptodactyly (52 cases, 4%)., Clinical Relevance: The OMT classification system is reasonably practical for precise classification of CULAs and enables easy comparison of studies over different time periods. However, continually updating the OMT classification system is required to better categorize the highly variable presentations of CULAs., (Copyright © 2023 American Society for Surgery of the Hand. Published by Elsevier Inc. All rights reserved.)

Guided bone regeneration surgery is an important dental operation used to regenerate enough bone to successfully heal dental implants. When this technique is performed on maxilla sinuses, hyaluronic acid (HLA) can be used as an auxiliary material to improve the graft material handling properties. Recent studies have indicated that low-molecular hyaluronic acid (L-HLA) provides a better regeneration ability than high-molecular-weight (H-HLA) analogues. The aim of this study was to fabricate an L-HLA-carboxymethyl cellulose (CMC) hybrid to promote bone regeneration while maintaining viscosity. The proliferation effect of fabricated L-HLA was tested using dental pulp stem cells (DPSCs). The mitogen-activated protein kinase (MAPK) pathway was examined using cells cultured with L-HLA combined with extracellular-signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38 inhibitors. The bone growth promotion of fabricated L-HLA/CMC hybrids was tested using an animal model. Micro-computer tomography (Micro-CT) and histological images were evaluated quantitatively to compare the differences in the osteogenesis between the H-HLA and L-HLA. Our results show that the fabricated L-HLA can bind to CD44 on the DPSC cell membranes and affect MAPK pathways, resulting in a prompt proliferation rate increase. Micro CT images show that new bone formation in rabbit calvaria defects treated with L-HLA/CMC was almost two times higher than in defects filled with H-HLA/CMC (p < 0.05) at 4 weeks, a trend that remained at 8 weeks and was confirmed by HE-stained images. According to these findings, it is reasonable to conclude that L-HLA provides better bone healing than H-HLA, and that the L-HLA/CMC fabricated in this study is a potential candidate for improving bone healing efficiency when a guided bone regeneration surgery was performed.

Background: Hepatic artery reconstruction (HAR) for liver transplantation is crucial for successful outcomes. We evaluated transplantation outcome improvement through continual technical refinements., Materials and Methods: HAR was performed in 1448 living donor liver transplants by a single plastic surgeon from 2008 to 2020. Difficult HARs were defined as graft or recipient hepatic artery ≤2 mm, size discrepancy (≥2 to 1), multiple hepatic arteries, suboptimal quality, intimal dissection of graft or recipient hepatic artery (HA), and immediate redo during transplantation. Technique refinements include early vessel injury recognition, precise HA dissection, the use of clips to ligate branches, an oblique cut for all HARs, a modified funneling method for size discrepancy, liberal use of an alternative artery to replace a pathologic HA, and reconstruction of a second HA for grafts with dual hepatic arteries in the graft., Results: Difficult HARs were small HA (21.35%), size discrepancy (12.57%), multiple hepatic arteries (11.28%), suboptimal quality (31.1%), intimal dissection (20.5%), and immediate redo (5.18%). The overall hepatic artery thrombosis (HAT) rate was 3.04% in this series. The average HAT rate during the last 4 years (2017-2020) was 1.46% (6/408), which was significantly lower than the average HAT rate from 2008 to 2016 (39/1040, 3.8%) with a statistical significance (p = 0.025). Treatment for posttransplant HAT included anastomosis after trim back (9), reconstruction using alternatives (19), and nonsurgical treatment with urokinase (9)., Conclusion: Careful examination of the HA under surgical microscope and selection of the appropriate recipient HA are key to successful reconstruction. Through continual technical refinements, we can reduce HA complications to the lowest degree., (Copyright © 2022 The Authors. Published by Elsevier Ltd.. All rights reserved.)

The molecular design of wide-bandgap conjugated polymer donors (WB-CPDs) is a promising strategy for tuning the bulk heterojunction blend film morphologies to achieve high-performance organic photovoltaic (OPV) devices. Herein, we synthesize two WB-CPDs, namely, PBQ-H and PBQ-M, with and without methyl groups on the fused-dithieno[3,2- f :2',3'- h ]quinoxaline (DTQx) moiety. We systematically investigate their structure-property relationship and OPV performances. The AFM and 2D grazing-incidence wide-angle X-ray scattering (GIWAXS) studies reveal that the PBQ-H:BO-4Cl BHJ blend shows strengthened aggregation behavior and stronger π-π stacking on face-on orientation compared with the PBQ-M:BO-4Cl BHJ blend, enhancing the phase separation, charge transport, and fill factor (FF). Blend film absorption spectra, however, show that the PBQ-H:BO-4Cl BHJ blend exhibits a lower absorption coefficient than that of the PBQ-M:BO-4Cl BHJ blend, which decreases the short-circuit current density ( J SC ). As a consequence, the optimized PBQ-H:BO-4Cl BHJ blend delivers a higher power conversion efficiency (PCE) of 12.88% with a J SC of 23.97 mA/cm 2 , an open-circuit voltage ( V OC ) of 0.86 V, and an FF of 62.46%, compared with the PBQ-M:BO-4Cl BHJ blend (PCE of 11.81% with a J SC of 24.78 mA/cm 2 , a V OC of 0.85 V, and an FF of 56.11%). Overall, this work demonstrates that alkyl group substitution on the DTQx moiety on the basis of WB-CPDs is critical for controlling the film morphology and thus obtaining high OPV performances.

Summary: The key to successful microvascular anastomosis is achieving intima-to-intima contact, which is not always easy. In this article, the authors propose the multiple-U technique, which is a novel microvascular anastomosis technique that characterizes easy and reliable intima-to-intima contact. The technique was performed on patients who underwent free flap reconstruction for head and neck defects at the Kaohsiung Chang Gung Memorial Hospital from September 1, 2020, to November 30, 2020. The immediate patency test results for all vessel anastomoses were positive, and the postoperative recovery courses of the patients were without any vascular complications. In conclusion, the multiple-U technique is a widely available technique that guarantees everted anastomosis sites and solid intima-to-intima contact. This technique can be performed on both arterial and venous anastomoses regardless of vessel size and wall thickness., (Copyright © 2022 by the American Society of Plastic Surgeons.)

Background: Intercellular cross-talking was suggested in matrix metalloproteinase (MMP)-9 expression with unknown mechanisms. Studies showed cyclophilin A (CypA) playing an important role in regulating MMP-9 expression in varied diseases. The aim of the study was to examine the CyPA on the MMP-9 augmentation in monocytic U937 cells after Porphyromonas gingivalis (Pg) lipopolysaccharide (LPS) treatment and human gingival fibroblast (hGF) co-culture., Methods: In independent culture or co-culture of hGF and U937 cell, quantitative real-time polymerase chain reaction (qPCR) and zymography were selected to examine the mRNA and protein activity of MMP-9, respectively. The CyPA expression was determined by qPCR., Results: LPS could enhance MMP-9 mRNA expression and enzyme activity in U937 cell. However, the enhancements were not observed in hGF. Similarly, LPS enhanced CyPA mRNA in U937, but not in hGF. After co-cultured with hGF, however, MMP-9 and CyPA in U937 increased regardless of the presence/absence of LPS. In U937 cells, the extra-supplied CyPA increased MMP-9 mRNA and enzyme activity, whereas the CyPA inhibitor, cyclosporine A, suppressed the LPS- and co-culture-enhanced MMP-9. Moreover, the inhibitors for MAP kinase, including PD98059 (ERK) and SP600125 (JNK), suppressed the CyPA-enhanced MMP-9 in U937., Conclusion: Through the CyPA pathway, the LPS and the hGF could augment the MMP-9 expression in the U937 cells., (© 2021 American Academy of Periodontology.)

Background/purpose: Culture environments play a critical role in stem cell expansion. This study aimed to evaluate the effects of 2,3,5,4'-tetrahydroxystilbene-2-O-b-D-glucoside (THSG) on the proliferation and differentiation of human dental pulp stem cells (DPSCs) in 2-dimensional (2D) and 3-dimensional (3D) culture systems., Materials and Methods: Human DPSCs were seeded in T25 flasks for 2D cultivation. For the 3D culture system, DPSCs were mixed with microcarriers and cultured in spinner flasks. Cells in both culture systems were treated with THSG, and cell proliferation was determined using a cell counter and a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide assay. In THSG-treated DPSCs, the genes associated with proliferation, adipogenesis, neurogenesis, osteogenesis, pluripotency, oncogenesis, and apoptosis were analyzed using real-time polymerase chain reactions., Results: The spinner flask time-dependently improved cell numbers, cell viability, and expansion rates in THSG-treated DPSCs. In both the T25 and spinner flasks, the messenger RNA (mRNA) levels of proliferation, osteogenesis, and pluripotent-related genes had a significant maximum expression with 10 μM THSG treatment. However, 0.1 μM of THSG may be the most suitable condition for triggering neurogenesis and adipogenesis gene expression when DPSCs were cultured in spinner flasks. Furthermore, the number of oncogenes and apoptotic genes decreased considerably in the presence of THSG in both the T25 and spinner flasks., Conclusion: The spinner flask bioreactor combined with THSG may upregulate proliferation and lineage-specific differentiation in DPSCs. Thus, the combination can be used to mass-produce and cultivate human DPSCs for regenerative dentistry., Competing Interests: The authors report no conflicts of interest related to this study., (© 2021 Association for Dental Sciences of the Republic of China. Publishing services by Elsevier B.V.)

Background/purpose: Immediate placement in the esthetic zone has been a predictable treatment option. However, it requires the clinician to be experienced and knowledgeable about esthetic diagnosis, accurate 3-dimensional (3D) implant placement, and restoratively driven planning/placement. Therefore, this study aimed to investigate a novel workflow integrating dynamic navigation to immediate single-implant placement in the aesthetic zone., Materials and Methods: We included ten patients who required at least one implant in the esthetic area and were treated with post-extraction socket implant placement. Osteotomy and implant placement followed computer-assisted implant positioning and image-guided dynamic navigation. Treatment outcomes were implant success rates, surgical and prosthetic complications, marginal bone level (MBL), modified pink esthetic score, and white score., Results: In the consecutive clinical cases, patients were satisfied with implant therapy's function and esthetic outcome in the esthetic zone. No other surgical or biological complications occurred, which accounts for the 100% cumulative success rate. The mean MBL was -0.76 ± 0.15 mm assessed using standardized intraoral digital periapical radiographs., Conclusion: The novel application of a dynamic guided navigation system is a dependable clinical protocol to obtain optimal implant position/angulation and esthetics on immediate implant placement., Competing Interests: The authors have no conflicts of interest relevant to this study., (© 2021 Association for Dental Sciences of the Republic of China. Publishing services by Elsevier B.V.)

Background/purpose: Based on the fundamental of the S3-level clinical practice guideline (CPG) for treating stage I-III periodontitis developed by the European Federation of Periodontology (EFP), this consensus report aimed to develop treatment recommendations for treating periodontitis in the Taiwanese population., Methods: The report was constructed by experts from the Taiwan Academy of Periodontology. The following topics were reviewed: (a) the prevalence of periodontitis in Asia and current status of treatment in Taiwan; (b) specific anatomical considerations for treating periodontitis in Asians; (d) educational and preventive interventions and supragingival plaque control; (d) subgingival instrumentation and adjunctive treatment; (e) surgical periodontal therapy; and (f) maintenance and supportive periodontal care. Recommendations were made according to the evidences from the EFP CPG, the published literature and clinical studies in Asians, and the expert opinions., Results: The treatment recommendations for the Taiwanese population were generally in parallel with the EFP CPG, and extra cautions during treatment and maintenance phases were advised due to the anatomical variations, such as shorter root trunk, higher prevalence of supernumerary distolingual root and lingual bony concavity in mandibular posteriors, and thinner anterior labial plate, of the Asian population., Conclusion: The EFP CPG could be adopted for treating periodontitis and maintaining periodontal health of the Taiwanese population, and anatomical variations should be cautious when the treatment is delivered., Competing Interests: Declaration of competing interest The authors have no conflicts of interest relevant to this article., (Copyright © 2021. Published by Elsevier B.V.)

Background: This study aimed to identify the potential circulating biomarkers of protein, mRNAs, and long non-coding RNAs (lncRNAs) to differentiate the papillary thyroid cancers from benign thyroid tumors., Methods: The study population of 100 patients was classified into identification (10 patients with papillary thyroid cancers and 10 patients with benign thyroid tumors) and validation groups (45 patients with papillary thyroid cancers and 35 patients with benign thyroid tumors). The Sengenics Immunome Protein Array-combined data mining approach using the Open Targets Platform was used to identify the putative protein biomarkers, and their expression validated using the enzyme-linked immunosorbent assay. Next-generation sequencing by Illumina HiSeq was used for the detection of dysregulated mRNAs and lncRNAs. The website Timer v2.0 helped identify the putative mRNA biomarkers, which were significantly over-expressed in papillary thyroid cancers than in adjacent normal thyroid tissue. The mRNA and lncRNA biomarker expression was validated by a real-time polymerase chain reaction., Results: Although putative protein and mRNA biomarkers have been identified, their serum expression could not be confirmed in the validation cohorts. In addition, seven lncRNAs (TCONS&#95;00516490, TCONS&#95;00336559, TCONS&#95;00311568, TCONS&#95;00321917, TCONS&#95;00336522, TCONS&#95;00282483, and TCONS&#95;00494326) were identified and validated as significantly downregulated in patients with papillary thyroid cancers compared to those with benign thyroid tumors. These seven lncRNAs showed moderate accuracy based on the area under the curve (AUC = 0.736) of receiver operating characteristic in predicting the occurrence of papillary thyroid cancers., Conclusions: We identified seven downregulated circulating lncRNAs with the potential for predicting the occurrence of papillary thyroid cancers., (© 2021. Italian Society of Endocrinology (SIE).)

Background/purpose: Space-making is one of the essential factors for bone regeneration in severe bony defect. To test the hypothesis that an appropriately designed scaffold may be beneficial for the bone formation in defect, the new bone formed in the critical-size calvarial defect of rat was examined after implanted with a 3D-printed poly-ɛ-caprolactone (PCL) scaffold, retaining with and without plasma rich fibrin (PRF)., Materials and Methods: Thirty-two rats were divided into four groups (control, PCL, PRF, and PCL-plus-PRF). A custom-made 3D-printed PCL scaffold, 900 μm in pore size, retaining with and without PRF, was implanted into a critical-sized calvarial defect, 6 mm in diameter. Animals were sacrificed at week-4 or 8 after implantation for assessing the new bone formation by dental radiography, micro-computed tomography (μ-CT), and histology., Results: By radiography and μ-CT, significantly greater mineralization areas/volumes were observed in defects with 3D-printed scaffold groups compared to that without the scaffold in both two-time points. However, no advantage was found by adding PRF. Histology showed that bone tissues grew into the central zone of the critical defect when 3D-printed PCL scaffold was present. In contrast, for the groups without the scaffolds, new bones were formed mostly along defect borders, and the central zones of the defects were collapsed and healed with thin connective tissue., Conclusion: Our results suggest that the use of a 900 μm pore size 3D-printed PCL scaffold may have the potential in facilitating the new bone formation., Competing Interests: The authors have no conflicts of interest relevant to this article., (© 2021 Association for Dental Sciences of the Republic of China. Publishing services by Elsevier B.V.)

Macrophages emerge in the milieu around innervated neurons after nerve injuries. Following nerve injury, autophagy is induced in macrophages and affects the regulation of inflammatory responses. It is closely linked to neuroinflammation, while the immunosuppressive drug tacrolimus (FK506) enhances nerve regeneration following nerve crush injury and nerve allotransplantation with additional neuroprotective and neurotrophic functions. The combined use of FK506 and adipose-derived stem cells (ADSCs) was employed in cell therapy for organ transplantation and vascularized composite allotransplantation. This study aimed to investigate the topical application of exosomes secreted by ADSCs following FK506 treatment (ADSC-F-exo) to the injured nerve in a mouse model of sciatic nerve crush injury. Furthermore, isobaric tags for relative and absolute quantitation (iTRAQ) were used to profile the potential exosomal proteins involved in autophagy. Immunohistochemical analysis revealed that nerve crush injuries significantly induced autophagy in the dorsal root ganglia and dorsal horn of the spinal segments. Locally applied ADSC-F-exo significantly reduced autophagy of macrophages in the spinal segments after nerve crush injury. Proteomic analysis showed that of the 22 abundant exosomal proteins detected in ADSC-F-exo, heat shock protein family A member 8 (HSPA8) and eukaryotic translation elongation factor 1 alpha 1 (EEF1A1) are involved in exosome-mediated autophagy reduction.

Statement of Problem: Restoring the edentulous maxilla with an implant-retained overdenture (IRO) can present a challenge because of increased implant divergence and the added wear of the implant abutments and attachments. However, knowledge pertaining to the degree of implant divergence and its effects on the wear pattern of the implant attachments is lacking., Purpose: The purpose of this in vitro study was to investigate the change in the retention and wear characteristics of unsplinted abutments and attachments when used to retain a maxillary 4-implant palateless complete removable overdenture with different implant angulations., Material and Methods: Three groups of specimens of 0-, 15-, and 30-degree implant angulations were evaluated, each with 7 specimens. The retention of specimens was recorded after aging cycles equivalent to 1, 2.5, and 6 years of use. The surface of the attachments was examined with light and scanning electron microscope (SEM)., Results: Increasing the interimplant divergence improved the initial retention. After 1 year of use, retention of the 15- and 30-degree groups was significantly higher than that of the 0-degree group (P<.05). No significant difference in retention was found between the 15- and 30-degree groups (P>.05). After 2.5 and 6 years of use, no significant difference in retention was noted between groups (P>.05). Under light and SEM examination, the wear of the abutments and attachments was related to the interimplant divergence., Conclusions: The initial retention of single attachments increased significantly as the implant divergence increased. The retention of the 15- and 30-degree groups was significantly higher than that of the 0-degree group after 1 year of use. No significant difference in retention was found after 2.5 and 6 years, regardless of implant angulation., (Copyright © 2021 Editorial Council for the Journal of Prosthetic Dentistry. Published by Elsevier Inc. All rights reserved.)

Exosomes secreted by adipose-derived stem cells (ADSCs) enhance angiogenesis and wound healing. However, in clinical settings, wounds may be infected by various bacteria or pathogens. We investigated whether human ADSCs stimulated with lipopolysaccharide (LPS) secrete exosomes (ADSC-LPS-exo) that augment the angiogenesis of human umbilical vein endothelial cells (HUVECs). ExoQuick-TC exosome precipitation solution was used to purify exosomes from human ADSC culture media in the presence or absence of 1 µg/mL LPS treatment for 24 h. The uptake of ADSC-LPS-exo significantly induced the activation of cAMP response element binding protein (CREB), activating protein 1 (AP-1), and nuclear factor-κB (NF-κB) signaling pathways and increased the migration of and tube formation in HUVECs. RNA interference with CREB, AP-1, or NF-κB1 significantly reduced the migration of and tube formation in HUVECs treated with ADSC-LPS-exo. An experiment with an antibody array for 25 angiogenesis-related proteins revealed that only interleukin-8 expression was significantly upregulated in HUVECs treated with ADSC-LPS-exo. In addition, proteomic analysis revealed that eukaryotic translation initiation factor 4E, amyloid beta A4 protein, integrin beta-1, and ras-related C3 botulinum toxin substrate 1 may be potential candidates involved in ADSC-LPS-exo-mediated enhanced angiogenesis.

Exosomes secreted by adipose-derived stem cells (ADSC-exo) reportedly improve nerve regeneration after peripheral nerve injury. Herein, we investigated whether pretreatment of ADSCs with FK506, an immunosuppressive drug that enhances nerve regeneration, could secret exosomes (ADSC-F-exo) that further augment nerve regeneration. Designed exosomes were topically applied to injured nerve in a mouse model of sciatic nerve crush injury to assess the nerve regeneration efficacy. Outcomes were determined by histomorphometric analysis of semi-thin nerve sections stained with toluidine blue, mouse neurogenesis PCR array, and neurotrophin expression in distal nerve segments. Isobaric tags for relative and absolute quantitation (iTRAQ) were used to profile potential exosomal proteins facilitating nerve regeneration. We observed that locally applied ADSC-exo and ADSC-F-exo significantly enhanced nerve regeneration after nerve crush injury. Pretreatment of ADSCs with FK506 failed to produce exosomes possessing more potent molecules for enhanced nerve regeneration. Proteomic analysis revealed that of 192 exosomal proteins detected in both ADSC-exo and ADSC-F-exo, histone deacetylases (HDACs), amyloid-beta A4 protein (APP), and integrin beta-1 (ITGB1) might be involved in enhancing nerve regeneration.

Osteoconduction is an important consideration for fabricating bio-active materials for bone regeneration. For years, hydroxyapatite and β-calcium triphosphate (β-TCP) have been used to develop bone grafts for treating bone defects. However, this material can be difficult to handle due to filling material sagging. High molecular weight hyaluronic acid (H-HA) can be used as a carrier to address this problem and improve operability. However, the effect of H-HA on bone formation is still controversial. In this study, low molecular weight hyaluronic acid (L-HA) was fabricated using gamma-ray irradiation. The viscoelastic properties and chemical structure of the fabricated hybrids were evaluated by a rheological analysis nuclear magnetic resonance (NMR) spectrum. The L-MH was mixed with H-HA to produce H-HA/L-HA hybrids at ratios of 80:20, 50:50 and 20:80 ( w / w ). These HA hybrids were then combined with hydroxyapatite and β-TCP to create a novel bone graft composite. For animal study, artificial bone defects were prepared in rabbit femurs. After 12 weeks of healing, the rabbits were scarified, and the healing statuses were observed and evaluated through micro-computer tomography (CT) and tissue histological images. Our viscoelastic analysis showed that an HA hybrid consisting 20% H-HA is sufficient to maintain elasticity; however, the addition of L-HA dramatically decreases the dynamic viscosity of the HA hybrid. Micro-CT images showed that the new bone formations in the rabbit femur defect model treated with 50% and 80% L-HA were 1.47 ( p < 0.05) and 2.26 ( p < 0.01) times higher than samples filled with HA free bone graft. In addition, a similar tendency was observed in the results of HE staining. These results lead us to suggest that the material with an H-HA/L-HA ratio of 50:50 exhibited acceptable viscosity and significant new bone formation. Thus, it is reasonable to suggest that it may be a potential candidate to serve as a supporting system for improving the operability of granular bone grafts and enhancing new bone formations.

Purpose: Although two ways of reconstruction have been reviewed for a long period, comparison of large sample size with nearly randomized patient selection is rare. Here, we compare the surgical outcomes of these two methods from two large medical institutes., Methods: Totally 176 patients were included from two medical centers with different protocols, 62 patients from MacKay Hospital repair defects with fibula flaps using reconstruction plates. Also 114 patients from Kaohsiung ChangGung Hospital treat with reconstruction plates combine anterior lateral thigh flaps, in which 12 cases had secondary fibula flap after plate exposure. Electronic medical records were reviewed retrospectively. Short (3-12 months) and long-term (>12 months) outcomes were evaluated., Results: Short-term evaluation revealed that the secondary fibula had a higher rate of vascular complications (33 vs. 6%, p = .02). Primary fibula had a higher risk of post reconstructive surgical debridement (60 vs. 40%, p = .018). Long-term follow-up revealed radiation therapy significantly worsened bone healing (p = .03). Primary fibula had a lower rate of screw loosening (14.0 vs. 40.3%, p = .002). Radiation therapy did not increase the plate exposure rate among the groups (33 vs. 27%, p = .389). Disease free survival showed no significant difference between two groups (53.1 ± 3.5 vs. 47.8 ± 2.8 months, p = .317)., Conclusion: Primary fibula reconstruction should be considered whenever possible to reduce the risk of late complications and simplifies the management after a potential plate exposure. Notably, radiation therapy apparently has no significant effect on plate exposure rate both in ALT or fibula groups., (© 2021 Wiley Periodicals LLC.)

Background: Excess lymphedematous tissue causes excessive oxidative stress in lymphedema. Lymphaticovenous anastomosis (LVA) supermicrosurgery is currently emerging as the first-line surgical intervention for lymphedema. No data are available regarding the changes in serum proteins correlating to oxidative stress and antioxidant capacity before and after LVA., Methods: A total of 26 patients with unilateral lower limb lymphedema confirmed by lymphoscintigraphy were recruited, and venous serum samples were collected before (pre-LVA) and after LVA (post-LVA). In 16 patients, the serum proteins were identified by isobaric tags for relative and absolute quantitation-based quantitative proteomic analysis with subsequent validation of protein expression by enzyme-linked immunosorbent assay. An Oxidative Stress Panel Kit was used on an additional 10 patients. Magnetic resonance (MR) volumetry was used to measure t limb volume six months after LVA., Results: This study identified that catalase (CAT) was significantly downregulated after LVA (pre-LVA vs. post-LVA, 2651 ± 2101 vs. 1448 ± 593 ng/mL, respectively, p = 0.033). There were significantly higher levels of post-LVA serum total antioxidant capacity (pre-LVA vs. post-LVA, 441 ± 81 vs. 488 ± 59 µmole/L, respectively, p = 0.031) and glutathione peroxidase (pre-LVA vs. post-LVA, 73 ± 20 vs. 92 ± 29 U/g, respectively, p = 0.018) than pre-LVA serum. In addition, after LVA, there were significantly more differences between post-LVA and pre-LVA serum levels of CAT (good outcome vs. fair outcome, -2593 ± 2363 vs. 178 ± 603 ng/mL, respectively, p = 0.021) and peroxiredoxin-2 (PRDX2) (good outcome vs. fair outcome, -7782 ± 7347 vs. -397 ± 1235 pg/mL, respectively, p = 0.037) in those patients with good outcomes (≥40% volume reduction in MR volumetry) than those with fair outcomes (<40% volume reduction in MR volumetry)., Conclusions: The study revealed that following LVA, differences in some specific oxidative stress markers and antioxidant capacity can be found in the serum of patients with lymphedema.

Objective: We investigated the importance of reactive oxygen species (ROS) on developing gingival overgrowth (GO) and then introduced the antioxidant strategy to prevent, or even reduce GO., Background: Gingival overgrowth is a common side effect of the patients receiving cyclosporine A (CsA), an immune suppressant. Although it has been broadly investigated, the exact pathogenesis of the induced GO is still uncertain., Methods: We cultured human primary gingival fibroblasts and used animal model of GO to investigate the ameliorative effects of antioxidants on CsA-induced GO. To examine the CsA-induced oxidative stress, associated genes and protein expression, and the overgrown gingiva of rats by using immunocytochemistry, confocal laser scanning microscopy, real-time PCR, ELISA, gelatin zymography, gingival morphological, and immunohistochemical analysis., Results: We found for the first time that ROS was responsible for the CsA-induced oxidative stress and TGF-β1 expression in human primary gingival fibroblasts, as well as the GO of rats. The antioxidants (oxidative scavenger of vitamin E and an antioxidative enzyme inducer of hemin) ameliorated CsA-induced pathological and morphological alterations of GO without affected the CsA-suppressed il-2 expression in rats. CsA-induced oxidative stress, HO-1, TGF-β1, and type II EMT were also rescued by antioxidants treatment., Conclusions: We concluded that CsA repetitively stimulating the production of ROS is the cause of CsA-GO which is ameliorated by treating antioxidants, including vitamin E and sulforaphane. Furthermore, the immunosuppressive effect of CsA is not interfered by antioxidant treatments in rats. This finding may thus help the clinician devise better prevention strategies in patients susceptible to GO., (© 2021 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Abstract., Background/purpose: Although 2,3,5,4'-Tetrahydroxystilbene-2-O-beta-glucoside (THSG) reportedly has anti-inflammatory properties, its role in inducing the dedifferentiation of human dental pulp stem cells (DPSC) into pluripotent-like stem cells remains to be determined. The purpose of this study is to evaluate the effects of THSG on the pluripotent-like possibility and mechanism of DPSC., Materials and Methods: DPSCs were treated with THSG, and cell viability was determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTS) assay. Real-time polymerase chain reaction was used to analyze the mRNA expression levels of pluripotency-associated genes and oncogenes and to detect telomerase activity in the cells. Embryoid body formation assay was conducted, and pluripotency-related proteins were identified using Western blotting. Data were analyzed using one-way analysis of variance., Results: Cell viability, telomerase activity, and embryoid body formation were enhanced in THSG-treated DPSCs. The mRNA expression levels of pluripotent-like genes (including Nanog homeobox [ NANOG ], SRY-box 2 [ SOX2 ], and POU class 5 homeobox 1 [ POU5F1/OCT4 ]) significantly increased after THSG treatment. The expression levels of pluripotency-related genes (Janus kinase-signal transducer 2 [ JAK2 ] and signal transducer and activator of transcription 3 [ STAT3 ]) increased, whereas those of oncogenes ( Ras , SRC , HER2 , and C-sis ) decreased. Furthermore, the expression levels of the phosphorylated JAK2 and STAT3 proteins significantly increased after THSG treatment., Conclusion: THSG treatment may enhance the pluripotent-like possibility of DPSC through the JAK2/STAT3 axis. Hence, it may be used as an alternative cell-based therapeutic strategy in regenerative dentistry., (© 2020 Association for Dental Sciences of the Republic of China. Publishing services by Elsevier B.V.)

Background: This study aimed to investigate the regenerative effects of 2,3,5,4'-tetrahydroxystilbene-2-O-β-d-glucoside (THSG)-treated human dental pulp stem cells (DPSC) on the healing of experimental periodontal defects in rats., Methods: The maxillary first molars of 30 male Sprague-Dawley rats were extracted, and after healing, bilateral periodontal defects were surgically created mesially in second molars. The defects were treated with Matrigel (as control), DPSC, or DPSC + THSG. After 2 weeks, the healed defects were evaluated using microcomputed tomography and through histological and immunohistochemical analyses., Results: In the microcomputed tomography analysis, more new bone formation in the DPSC and DPSC + THSG groups was observed compared with the control group. The periodontal bone supporting ratio in site with DPSC + THSG was significantly higher than that in DPSC. Histologically, an enhanced new bone formation and more significant periodontal attachment were observed in the DPSC + THSG group. The expression levels of proliferating cell nuclear antigen (PCNA), vascular endothelial growth factor (VEGF), and osteopontin (OPN) in the DPSC + THSG group were significantly greater than those in other groups., Conclusions: THSG-revolutionized DPSCs significantly shortened the regenerative period of periodontal defects by enhancing the cell recruitment and possibly the angiogenesis in rat models, which illustrate the critical implications for a clinical application and provide a novel tactic for periodontitis treatment., (© 2020 American Academy of Periodontology.)

The heterogeneity of exosome populations presents a great challenge to their study. The current study was designed to investigate the potential heterogeneity miRNA contents in circulating exosomes purified via different exosomal markers. In this study, exosomes from the serum of C57BL/6 mice after cecum ligation and perforation (CLP) or sham operation were isolated by precipitation using ExoQuick-TC and affinity purified with anti-Rab5b, anti-CD9, anti-CD31, and anti-CD44 antibodies using the Exo-Flow Exosome Capture kit to collect exosome subpopulations. RNA extracted from the exosomes isolated by ExoQuick-TC were profiled by next-generation sequencing (NGS). Real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) was also employed to determine the expression profiles of four representative exosomal miRNAs (mmu-miR-486-5p, mmu-miR-10a-5p, mmu-miR-143-3p, and mmu-miR-25-3p) selected from the NGS analysis. The results revealed that the expression patterns of these miRNAs in exosomes isolated by ExoQuick-TC as determined by RT-qPCR and NGS were similar, showing upregulation of mmu-miR-10a-5p and mmu-miR-143-3p but downregulation of mmu-miR-25-3p and mmu-miR-486-5p following CLP when compared to the levels in exosomes from sham control mice. However, their expression levels in the antibody-captured exosome subpopulations varied. The miRNAs in the exosomes captured by anti-Rab5b or anti-CD9 antibodies were more similar to those isolated by ExoQuick-TC than to those captured by anti-CD44 antibodies. However, there were no significant differences in these four miRNAs in CD31-captured exosomes. This study demonstrated that purification with different exosomal markers allows the collection of different exosome subpopulations with various miRNA contents. The results of this study demonstrate the heterogeneity of circulating exosomes and suggest the importance of stratifying exosome subpopulations when using circulating exosomes as biomarkers or investigating exosome function. In addition, this study also emphasized the necessity of using a consistent exosome marker across different samples as detecting biomarkers., Competing Interests: Competing Interests: The authors have declared that no competing interest exists., (© The author(s).)

Objective: This in vitro study aimed to evaluate the expression of cyclophilin A (CyPA) in U937 monocytic cells after coculturing with the human gingival fibroblasts (HGFs) and the effect of CyPA on the augmentation of MMP-2 expression in the coculture environment., Background: Leukocyte infiltration in gingival connective tissue is one of the major findings in the lesions of inflammatory periodontal diseases. A crosstalk between the resident gingival fibroblasts and the recruited inflammatory cells that promote the expression of matrix metalloproteinases (MMPs) was proposed based on recent findings, whereas the cluster of differentiation 147 (CD147)-CyPA pathway was suggested to be involved with the crosstalk., Material and Methods: CyPA was released into media, in the independent or transwell coculture of HGF and U937 cells, as determined by enzyme-linked immunosorbent assay, whereas intracellular mRNA expressions for CyPA and MMP-2 were examined by quantitative real-time polymerase chain reaction, in the transwell coculture or conditional medium models. Zymography was conducted to analyze the activities of pro-MMP-2/MMP-2 released into the media., Results: (a) A significantly increased CyPA protein level was observed in the transwell coculture media compared with that in the independent culture. (b) The transwell coculture-enhanced mRNA expression for CyPA was noticed in U937 cells but not in HGFs. After adding with HGF-conditioned medium, the mRNA enhancement in U937 cells occurred in a dose-dependent manner. (c) Although the MMP-2 activities significantly increased after transwell coculturing, the MMP-2 mRNA enhancement was observed only in HGFs. (d) Exogenous CyPA could enhance MMP-2 activities in HGFs in a dose-dependent manner. However, the CyPA antagonist reduced the MMP-2 activities in the transwell cocultures. (e) Moreover, the CyPA-enhanced MMP-2 activity in HGF was decreased significantly by the pathway inhibitor for c-Jun amino-terminal kinase (JNK)., Conclusion: Based on the present findings, we suggest that gingival fibroblasts could enhance the CyPA release from U937 cells, via the JNK pathway, resulting in MMP-2 enhancement in fibroblasts. The finding shed light on a new mechanism of cellular interaction involving MMP-2 and CyPA, in two cells., (© 2020 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Background: From the perspective of a surgeon, knowledge of the operative risk factors that may affect postoperative outcomes is important in free anterolateral thigh (ALT) flap reconstruction for head and neck defects after tumor ablation. Therefore, this study was designed to examine the surgical intervention factors related to postoperative complications in a propensity score matched patient population., Patients and Methods: A total of 1,284 head and neck cancer patients who received free ALT flap repair over a 9-year period from March 1, 2008, to February 28, 2017, at a single medical center were selected and divided into two groups (without complications, n = 845 and with complications, n = 439) according to the presence or absence of complications at the recipient site. Complications were defined as the detection of hematoma, surgical site infection, partial flap loss, oral fistula formation, flap partial necrosis, and flap loss. Well-balanced propensity score-matched cohorts with 292 patients each were created using the 1:1 Greedy algorithm, with adjustment for significant baseline patient characteristics., Results: The patients with postoperative complications had a higher proportion of individuals with betel nut chewing (91.8% vs. 86.6%, p = .008), diabetes mellitus (23.0% vs. 17.8%, p = .029), and preoperative chemotherapy (31.7% vs. 25.3%, p = .019), and higher serum creatinine levels (median [Q1-Q3]: 0.92 [0.80-1.07] vs. 0.89 [0.77-1.06], p = .008) and lower serum albumin levels (4.2 [3.9-4.5] vs. 4.3 [4.1-4.5], p < .001) than those without postoperative complications. Individual operator (p < .001), the length of flap (20 [15-23] cm vs. 20 [15-25] cm, p < .001), operative time (6.9 hr [5.7-8.3 hr] vs. 7.3 hr [5.9-8.7 hr], p = .001), operation start time (p = .003), and units of transfused packed red blood cells (0.0 [0.0-0.0] units vs. 0.0 [0.0-2.0] units, p < .001) were the factors significantly associated with the occurrence of postoperative complications. However, in the matched patient cohorts, individual operator (p = .003), flap length (18 [15-22] cm vs. 20 [15-25] cm, p < .001) and length-to-width ratio (2.6 [2.0-3.3] vs. 3.0 [2.5-3.6], p < .001), and operative time (6.9 hr [5.7-8.3 hr] vs. 7.2 hr [5.9-8.7 hr], p = .019) were associated with the occurrence of postoperative complications, but the operation start time (p = .285) and units of transfused packed red blood cells (p = .917) were not., Conclusions: This study demonstrated in matched patient cohorts that individual operator, flap size, and operative time were associated with postoperative complications of free ALT flap reconstruction in patients with head and neck cancer. To reduce the postoperative complication rate, this study implies the importance of length and length-to-width ratio in harvesting the flap, and meanwhile the surgeon experience in free-flap reconstruction., (© 2020 Wiley Periodicals, Inc.)

Background/ Purpose: The connective tissue graft from hard palate is a reliable graft technique that has been used for achieving root coverage, increasing keratinized tissue width and thickness in periodontal plastic surgeries. Donor site morbidities, including complications from postoperative bleeding, pain during healing phase, difficulties in eating and speaking and unexpecting healing patterns, are always a concern for both doctors and patients. The aim of this study was to investigate a novel design of palatal stent to reduce these complications and provide patient with a smooth healing experience after soft tissue harvesting from hard palate., Materials and Methods: Eight patients requiring connective tissue graft from palatal site were included in the study. The palatal stents made with light-curing hybrid composite resin were fabricated and tried in for patients prior to the periodontal plastic surgeries. Stent was delivered immediately without other dressing material or suture after the graft harvesting procedure for blood clot stabilization. Bleeding tendency was evaluated at the completion of the procedure. Patients came back for follow up in 1 week, 2 weeks and 1 month after the surgery. In the consecutive clinical cases, all patients reported minimal postoperative pain and discomfort (score ranged between 0 and 2). Both chewing and swallowing were not affected when wearing the stent, while four patients reported speaking inconvenience influenced by the stent., Results: In the consecutive clinical cases, all patient reported minimal postoperative pain and discomfort (Score ranged between 0 and 2). Both chewing and swallowing were not affected when wearing the stent while four patients reported speaking influenced by the stent., Conclusion: The novel design of palatal stent reduces donor site morbidity and provides patient with a good healing experience after soft tissue harvesting from hard palate., Competing Interests: The authors have no conflicts of interest relevant to this study., (© 2020 Association for Dental Sciences of the Republic of China. Publishing services by Elsevier B.V.)

Background: Cluster of differentiation 147 (CD147) is a multifunctional glycoprotein that functions as an inducer of matrix metalloproteinase (MMP) expression in fibroblasts. Synergistically enhanced MMP-2 expression was recently observed in the coculture of human gingival fibroblasts (HGFs) and U937 human monocytic cells; however, the responsible mechanisms have not yet been fully established. The aim of this study was to evaluate the release of soluble CD147 in HGFs after coculturing with U937 cells and its functional effect on the enhancement of MMP-2 expression in HGFs., Methods: Enzyme-linked immunosorbent assay was used to determine the amount of CD147 protein in media, whereas real-time polymerase chain reaction was performed to evaluate the mRNA levels of CD147 and MMP-2 in HGFs and U937 cells. The enzyme activities of MMP-2 released from cells were examined by zymography. Transwell coculturing and conditioned media treatments were selected to rule out the effect of direct contact of HGFs and U937 cells., Results: The protein and mRNA expression of CD147 in HGFs were enhanced after transwell coculturing with U937 cells and exposure to U937-conditioned medium. MMP-2 enzyme activities in HGFs were also significantly increased by the coculturing methods. Administration of exogenous CD147 enhanced MMP-2 expression in HGFs, whereas treatment with cyclosporine-A, which inhibited CD147 expression, reduced U937-enhanced MMP-2 expression in HGFs., Conclusions: CD147 can interact with fibroblasts to stimulate the expression of MMPs associated with periodontal extracellular matrix degradation. This study has demonstrated that CD147 released from fibroblasts might play a role in monocyte-enhanced MMP-2 expression in HGFs., (© 2019 American Academy of Periodontology.)

Background: Bile duct complications hamper patient recovery from a transplant, curtail patient quality of life, and may impair the function and persistent survival of the allograft. The aim of this study is to compare the results of biliary reconstruction using 6-0 polypropylene and 6-0 polydioxanone., Methods: A single-center retrospective study was performed on 133 patients who underwent microsurgical biliary reconstruction during living donor liver transplantation between November 2014 and December 2015. 6-0 polypropylene (Prolene) and 6-0 polydioxanone suture (PDS) were used for biliary reconstruction in 80 and 53 cases, respectively. The factors of age, sex, disease, and comorbidity were evenly distributed in these 2 groups. The average follow-up time was 43 months (36-50 months). There were 49 right lobe grafts and 31 left lobe grafts in the polypropylene group and 27 right lobe grafts, 25 left lobe grafts, and 1 left lateral segment in the polydioxanone group., Results: The overall biliary complication rate was 11.25% in the polypropylene group. The overall biliary complication rate was 11.32% in the polydioxanone group. All biliary complications were managed successfully, and no mortality was observed. There was no statistically significant difference between complication rates with the use of different suture material (P = .990)., Conclusion: The theoretical advantages of polydioxanone over polypropylene in biliary reconstruction could not be explained with this study., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Objective: Efforts have been devoted to clarify the possible factors related to postoperative complications in free-flap reconstruction. While patient-related factors have been widely discussed, studies regarding the operation/operator-related factors are rather limited in the literature. This study was designed to investigate the relationship between operation/operator-related factors and the surgical complications in free-flap reconstruction following head and neck cancer resection., Methods: Data of 1,841 patients with a total of 1,865 free-flap reconstructions (24 double free-flap reconstructions) between March 2008 and February 2017 were retrieved from the registered microsurgery database of the hospital. The association of operation/operator-related factors (including flap length and length-width ratio, flap types, use of vein graft, opposite side microanastomosis, number of microanastomoses, operators, operator experience, and operation time) with surgical complications was assessed by 1:1 propensity score-matched study groups., Results: After propensity score matching of the patient-related factors, the rate of vein grafting was significantly higher (0.6% vs. 2.2%, p = .038) and the operation time was longer (7.0 [5.8-8.5] vs. 7.4 [6.1-8.8] hr, p = .006) in the complication group. In addition, flap length and length-width ratio, flap types, opposite side microanastomosis, number of microanastomoses, operators, and operator experience were not associated with surgical complications., Conclusions: In a hospital that consisted of surgeons with high-volume or very-high-volume experience, the operators or operation experience were not significantly associated with the surgical complications. Only a longer operation time was associated with surgical complications in the patients who underwent free-flap reconstruction for head and neck cancer., (© 2019 Wiley Periodicals, Inc.)

Background/purpose: Immediate implant placement has been considered to be a successful treatment procedure. The bone plate perforation (BPP) may be one of severe complication and potentially life-threatening situation. The aim of this virtual study is to evaluate the influences of angled implant insertion on BPP during immediate implant installation in the posterior mandible., Materials and Methods: Cone beam computed tomography images of 488 posterior teeth from 61 patients were selected. Virtual immediate implant placement (VIIP) was performed at each posterior tooth following the appropriate axis with the prosthetic-driven planning and different deviation angles of 3-, 6-, or 9-degree. BPP was then examined from cross-sectional images obtained. Furthermore, the relation of lingual bony morphology and BPP were also determined., Results: The incidence of buccal and lingual BPP increased as the deviation angle increased in posterior mandible area. Incidence of lingual BPP was significantly influenced by angular deviation and type of lingual bony morphology after adjusting for age, gender, tooth type, and right/left side. An increase in incidence odds of over 6-fold (OR = 6.583) was noted for placements angled by 9° compared with placements made without angulation, and an increase in incidence odds of over 3-fold (OR = 3.622) was noted for teeth with the undercut-type lingual morphology compared with the other types., Conclusion: The present Results indicate that accurate selection of the implant insertion angle and full awareness of the bony anatomy at the implant recipient site are essential to prevent BPP in the posterior mandible.

Objective: To establish the composition of bacteria in mice following cecum ligation and puncture (CLP) through metagenomic analysis and investigate the role of TLRs on the composition of bacteria., Methods: Total DNA extraction was done from the ascites, blood, and fecal samples from C57BL/6 mice sacrificed at 0, 4, 8, and 16 h, as well as from Tlr2-/-, Tlr4-/-, Tlr5-/-, and NF-κB-/-mice sacrificed at 16 h following CLP. Amplification of the V3-V4 regions of the bacterial 16S rRNA genes by PCR and the Illumina MiSeq sequencer was used for deep sequencing. Hierarchical clustering of the isolates was performed with Ward's method using Euclidean distances. The relative abundance according to operational taxonomic unit (OTU) number or taxa was used to compare the richness among subgroups in the experiments., Results: There were 18 taxa that had significantly different abundances among the different samples of the C57BL/6 mice at 16 h following CLP. Various dynamic changes in the infectious bacteria inside the peritoneal cavity after CLP were found. While knockout of Tlr5 and NF-κB impaired the ability of bacterial clearance inside the peritoneal cavity for some kinds of bacteria found in the C57BL/6 mice, the knockout of Tlr4 enhanced clearance for other kinds of bacteria, and they presented excessive abundance in the peritoneal cavity despite their scarce abundance in the stool., Conclusion: NF-κB and TLRs are involved in bacterial clearance and in the expression pattern of the bacterial abundance inside the peritoneal cavity during polymicrobial infection., Competing Interests: The authors have declared that no competing interests exist.

Background/purpose: To evaluate the measurement accuracy of hard-tissue thicknesses adjacent to dental implants with different thread designs on images obtained from cone beam computed tomography (CBCT) using an in vitro model., Materials and Methods: On 4 × 13-mm implant, the neck of the implant was designed with micro-threads, and the apical part was covered by macro-threads; these implants were placed in a vinyl polysiloxane block that mimicked hard-tissue. Models were prepared with various thicknesses of 2.0, 1.0, 0.5 and 0.3 mm adjacent to the dental implant. Each model was scanned using CBCT, and the thickness of the cortical bone from the outer surface of the micro-threads and macro-threads were recorded. Ground sections were prepared, and the thickness was measured with electronic calipers as the gold standard (GS) measurement., Results: CBCT measurements of the micro-thread surface were consistently underestimated compared to the GS measurement when the thickness of the hard-tissue-mimicking material was ≤1.0 mm. In comparison, CBCT measurements of the macro-thread surface closely approximated the standard measurement, except when the thickness of the hard-tissue-mimicking material was 0.3 mm. The mean percentage errors from the standard measurement for the 2.0-, 1.0-, 0.5-, and 0.3-mm thickness groups were 4.8%, 16.4%, 37.8%, and 92.6%, respectively, for the micro-thread group, and were 0.6%, 2.9%, 9.5%, and 40.8%, respectively, for the macro-thread group., Conclusion: Within the limitations of this study, we conclude that CBCT may not produce sufficient resolution for thin sections of hard tissue-mimicking materials adjacent to micro-thread surfaces.

Background: Prolonged static postures (PSPs) may predispose dentists to develop cervical herniated intervertebral disc (C-HIVD); however, there is limited evidence supporting this in the literature thus far. We conducted this study to fit the data gap., Methods: We conducted a retrospective nationwide population-based study using the Taiwan National Health Insurance Research Database to identify 10,930 dentists, an identical number of age- and sex-matched participants from the general population, and 73,718 other health care providers (HCPs, non-dentists). Comparisons for the risk of developing C-HIVD between dentists and the general population, and between dentists and other HCPs were performed by tracing their medical histories between 2007 and 2011., Results: Dentists had a cumulative incidence rate of 1.1% for C-HIVD during the 5-year follow-up period. Overall, there was no difference of the risk for C-HIVD between dentists and the general population after adjusting for hypertension, hyperlipidemia, liver disease, mental disorders, diabetes mellitus, coronary artery disease, chronic obstructive pulmonary disease, malignancy, stroke, and renal disease (adjusted odds ratio [AOR]: 1.2, 95% confidence interval [CI]: 0.9-1.6). However, stratified analysis showed that younger dentists (≤ 34 years) had a trend of higher risk for C-HIVD than members of the younger general population (AOR: 1.9, 95% CI: 0.9-4.1). There was no difference found between dentists and other HCPs (AOR: 0.9, 95% CI: 0.8-1.1)., Conclusion: Younger dentists had a trend of higher risk of developing C-HIVD than members of the general population.

Skin and soft tissue defect of the ankle usually mandates flap reconstruction; however, bulky flap and difficulty ambulance have been the major disadvantages after flap reconstruction of this area. We used the concept of full-thickness skin graft as a 1-stage method of secondary debulking procedure to achieve both aesthetic and functional results., Methods: Since January of 2004 to June of 2016, 22 one-stage secondary debulking procedures were performed on 22 patients who had received reconstruction with flaps for ankle defects. Nineteen cases were free myocutaneous flaps, 2 cases were free fasciocutaneous flaps, and 1 case was a distally based sural artery flap. In the operative technique, the full-thickness skin was harvested from the flap and regrafted on the defatted fascia with tie-over dressing. The functional and cosmetic outcomes as well as complications were reviewed., Results: The overall mean follow-up time was 12 months. After the debulking procedure, all of the grafted skins took well. The patients were able to wear their own shoes without difficulty and regained ease of ambulation. The reconstructed area was found to decrease to an average of 28.92% as compared with predebulking area. The reconstructed ankles achieve good symmetry with regard to the contralateral side (P < 0.05). All of the patients were satisfied with the results of the reconstructed ankles., Conclusions: The 1-stage secondary debulking procedure is a safe and reproducible technique that achieves good functional and aesthetic outcomes after flap reconstruction of the ankle.

Introduction: Although the therapeutic potential of human dental pulp stem cells (hDPSCs) has been studied for bone regeneration, the therapeutic efficiency needs further consideration and examinations for clinical applications. Thus, the aims of this study were to evaluate the effect of 2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucoside (THSG) on the osteogenic differentiation of hDPSCs and to examine the therapeutic efficiency of the THSG-enhanced osseous potential of hDPSCs in alveolar bony defects of rats., Methods: Expressions of osteogenic messenger RNAs (including ALP, RUNX2, BGLAP, and AMBN) were examined by quantitative real-time polymerase chain reaction. Alizarin red S staining was conducted to analyze THSG-induced mineralization of hDPSCs. To investigate the regenerative effects of THSG-treated hDPSCs on dental alveolar bone, bony defects were created in male Sprague-Dawley rats. Defects were treated with Matrigel (Corning Inc, Corning, NY), hDPSCs, or hDPSCs + THSG. After 2 weeks, defect healing was evaluated by micro-computed tomographic and histologic analyses., Results: In the cell model, THSG induced osteogenesis-associated genes (ALP, RUNX2, and BGLAP) and an enamel-related gene (AMBN), resulting in mineralization as detected by alizarin red S staining after 2 weeks of treatment. In the animal model, THSG increased all parameters of bone formation (the relative bone volume, trabecular thickness, trabecular number, and trabecular separation) in alveolar bony defects of rats. THSG not only improved the quality of newly formed bone but also the quantity of new bone., Conclusions: These results showed important findings in revealing the THSG-enhanced osteogenic differentiation of hDPSCs and THSG-facilitated bone regeneration, which may provide an alternative option for cell-based regenerative therapy., (Copyright © 2019 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

Background: This study evaluated the ameliorative effect of hesperidin (HES), an anti-inflammatory flavanone, in rats with ligation (Lig)-induced periodontitis., Methods: A total of 48 rats were randomly divided into non-ligation group (NL), Lig group, and two ligation-plus-HES groups (L+H). HES was administered immediately after ligature placement at a dose of 75 or 150 mg/kg by intragastric feeding. Destruction of the ligated maxillary second and mandibular first molars were evaluated by dental radiography, microcomputed tomography (micro-CT), and histometry performed after sacrificing the rats on the seventh day. The expression levels of interleukin (IL)-1β, IL-6, and inducible nitric oxide synthase (iNOS) messenger (m)RNAs in the gingiva were determined by reverse-transcription polymerase chain reaction. The expression of iNOS was examined by immunohistochemistry., Results: The dental radiography and micro-CT findings revealed significantly increased alveolar bone loss in the Lig group, which was significantly prevented by HES. The histometry results revealed less gingival inflammation and connective tissue loss in the L+H groups compared with that in the Lig group. The mRNA expression levels of IL-6, IL-1 β, and iNOS were significantly increased in the Lig group but were reduced in the L+H groups. The immunostaining results showed that the ligation-induced iNOS expression was also decreased by HES., Conclusions: Oral administration of HES promotes an ameliorative effect against the ligation-induced alveolar bone loss and effectively inhibits the production of proinflammatory mediators in rats with experimentally induced periodontitis. Therefore, HES may be a good candidate for modulating oral inflammatory diseases., (© 2018 American Academy of Periodontology.)

Background: The immediate challenges during microvascular reconstruction of hepatic artery (HAR) during liver transplantation (LT) can be many. Hence, in order to give a cross sectional view of these problems this study over a period of 1 year, showing our routine practice, was taken up., Methods: From January 2015 to December 2015, a total of 133 LTs were performed in Kaohsiung Chang Gung Memorial Hospital, Taiwan. All hepatic artery (HA) reconstructions were performed by a microvascular surgeon under an operating microscope., Results: In the 133 patients, one artery was anastomosed in 123 (92.5%) patients, two in 9 (6.8%) patients and three in 1 (0.7%) of the patient. Eleven (8.3%) arteries were less than 2 mm in size (1-1.9 mm). There were intimal dissections (IDs) involving either the donor or the recipient arteries of mild to severe nature in 9 (6.8%) patients. Immediately following graft arterial anastomosis, either there was no flow or an intraoperative hepatic artery thrombosis (HAT) was found in nine (7.1%-8 LDLT, 4.8%-1 DDLT) patients. Immediate re-do anastomosis was done in all of these patients who did well in the follow-up. The overall post-operative success rate was 99.2%. One patient (0.8%) developed postoperative HAT due to infection during follow up and died due to sepsis., Conclusions: Small vessels or HA injury are the frequently encountered problems by a micro vascular surgeon. The other problems could be ID, need to do multiple reconstructions, immediate HAT and ability to re-do the HAR immediately., Competing Interests: Conflicts of Interest: The authors have no conflicts of interest to declare.

Background: The purpose of this study was to build a model of machine learning (ML) for the prediction of mortality in patients with isolated moderate and severe traumatic brain injury (TBI)., Methods: Hospitalized adult patients registered in the Trauma Registry System between January 2009 and December 2015 were enrolled in this study. Only patients with an Abbreviated Injury Scale (AIS) score ≥ 3 points related to head injuries were included in this study. A total of 1734 (1564 survival and 170 non-survival) and 325 (293 survival and 32 non-survival) patients were included in the training and test sets, respectively., Results: Using demographics and injury characteristics, as well as patient laboratory data, predictive tools (e.g., logistic regression [LR], support vector machine [SVM], decision tree [DT], naive Bayes [NB], and artificial neural networks [ANN]) were used to determine the mortality of individual patients. The predictive performance was evaluated by accuracy, sensitivity, and specificity, as well as by area under the curve (AUC) measures of receiver operator characteristic curves. In the training set, all five ML models had a specificity of more than 90% and all ML models (except the NB) achieved an accuracy of more than 90%. Among them, the ANN had the highest sensitivity (80.59%) in mortality prediction. Regarding performance, the ANN had the highest AUC (0.968), followed by the LR (0.942), SVM (0.935), NB (0.908), and DT (0.872). In the test set, the ANN had the highest sensitivity (84.38%) in mortality prediction, followed by the SVM (65.63%), LR (59.38%), NB (59.38%), and DT (43.75%)., Conclusions: The ANN model provided the best prediction of mortality for patients with isolated moderate and severe TBI., Competing Interests: The authors have declared that no competing interests exist.

Background : For elderly trauma patients, a prognostic tool called the Geriatric Trauma Outcome Score (GTOS), where GTOS = (age) + (ISS × 2.5) + (22 if any packed red blood cells (pRBCs) were transfused within 24 h after admission), was developed for predicting mortality. In such calculation, a score of 22 was added in the calculation of GTOS regardless of the transfused units of blood. This study aimed to assess the effect of transfused blood units on the mortality outcomes of the elderly trauma patients who received blood transfusion (BT). Methods : Detailed data of 687 elderly trauma patients aged ≥65 years who were transfused with pRBCs within 24 h after admission into a level I trauma center between 1 January 2009 and 31 December 2016 were retrieved from the Trauma Registry System database. Based on the units of pRBCs transfused, the study population was divided into two groups to compare the mortality outcomes between these groups. Adjusted odds ratios (AORs) with its 95% confidence intervals (CIs) for mortality were calculated by adjusting sex, pre-existing comorbidities, and GTOS. Results : When the cut-off value of BT was set as 3 U of pRBCs, patients who received BT ≥ 3 U had higher odds of mortality than those who received BT < 3 U (OR, 3.0; 95% CI, 1.94⁻4.56; p < 0.001). Patients who received more units of pRBCs still showed higher odds of mortality than their counterparts. After adjusting for sex, pre-existing comorbidities, and GTOS, comparison revealed that the patients who received BT of 3 U to 6 U had a 1.7-fold adjusted odds of mortality than their counterparts. The patients who received BT ≥ 8 U and 10 U had a 2.1-fold (AOR, 2.1; 95% CI, 1.09⁻3.96; p < 0.001) and 4.4-fold (AOR, 4.4; 95% CI, 2.04⁻9.48; p < 0.001) adjusted odds of mortality than those who received BT < 8 U and <10 U, respectively. Conclusions : This study revealed that the units of BT did matter in determining the probability of mortality. For those who received more units of blood, the mortality may be underestimated according to the GTOS.

Background: The purpose of this study was to profile the bacterium in the ascites and blood of patients with acute surgical abdomen by metagenome analysis., Methods: A total of 97 patients with acute surgical abdomen were included in this study. Accompanied with the standard culture procedures, ascites and blood samples were collected for metagenome analysis to measure the relative abundance of bacteria among groups of patients and between blood and ascites., Results: Metagenomic analysis identified 107 bacterial taxa from the ascites of patients. A principal component analysis (PCA) could separate the bacteria of ascites into roughly three groups: peptic ulcer, perforated or non-perforated appendicitis, and a group which included cholecystitis, small bowel lesion, and colon perforation. Significant correlation between the bacteria of blood and ascites was found in nine bacterial taxa both in blood and ascites with more than 500 sequence reads. However, the PCA failed to separate the variation in the bacteria of blood into different groups of patients, and the bacteria of metagenomic analysis is only partly in accordance with those isolated from a conventional culture method., Conclusion: This study indicated that the metagenome analysis can provide limited information regarding the bacteria in the ascites and blood of patients with acute surgical abdomen.

Background: This study aimed at assessing the effect of a low-fat diet (LFD) in obese mice lacking toll⁻like receptors (Tlr) and understanding the expression and regulation of microRNAs during weight reduction. Methods: C57BL/6, Tlr5 -/- , Tlr2 -/- and Tlr4 -/- mice were used in this study. A group of mice were fed with a high-fat diet (HFD) (58% kcal) for 12 weeks to induce obesity (diet-induced obesity, DIO). Another group that had been fed with HFD for eight weeks (obese mice) were switched to a low-fat diet (LFD) (10.5% kcal) for the next four weeks to reduce their body weight. The control mice were fed with a standard AIN-76A diet for the entire 12 weeks. The body weight of the mice was measured weekly. At the end of the experiment, epididymal fat weight and adipocyte size were measured. The differentially expressed miRNAs in the fat tissue was determined by next-generation sequencing with real-time quantitative reverse transcription polymerase chain reaction (RT⁻qPCR). Target prediction and functional annotation of miRNAs were performed using miRSystem database. Results: Switching to LFD significantly reduced the body weight and epididymal fat mass in the HFD-fed C57BL/6 and Tlr5 -/- mice but not in Tlr2 -/- and Tlr4 -/- mice. Weight reduction significantly decreased the size of adipocytes in C57BL/6 but not in the Tlr knockout mice. In Tlr2 -/- and Tlr4 -/- mice, feeding with HFD and the subsequent weight reduction resulted in an aberrant miRNA expression in the epididymal fat tissue unlike in C57BL/6 and Tlr5 -/- . However, target prediction and functional annotation by miRSystem database revealed that all the top 10 Kyoto Encyclopedia of Genes and Genomes (KEGG) database pathways of the dysregulated miRNAs during weight reduction in the C57BL/6 mice were also found in the regulated pathways of Tlr5 -/- , Tlr2 -/- , and Tlr4 -/- strains. However, among these pathways, gene sets involved in arginine and proline metabolism and glutathione metabolism were mainly involved in the Tlr knockout mice but not in the C57BL/6 mice. Conclusions: In this study, we demonstrated that feeding of LFD leads to significant body weight reduction in C57BL/6 and Tlr5 -/- mice, but not in Tlr2 -/- and Tlr4 -/- mice. Significant reduction in the size of adipocytes of epididymal fat was only found in C57BL/6, but not in Tlr5 -/- , Tlr2 -/- , and Tlr4 -/- mice. The dysregulated miRNAs in Tlr2 -/- and Tlr4 -/- mice were different from those in C57BL/6 and Tlr5 -/- strains. Among those miRNA-regulated pathways, arginine and proline metabolism as well as glutathione metabolism may have important roles in the Tlr knockout mice rather than in C57BL/6 mice.