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Abstract Cancers with activating mutations of KRAS show a high prevalence but remain intractable, requiring innovative strategies to overcome the poor targetability of KRAS. Here, we report that KRAS expression is post-translationally up-regulated through deubiquitination when the scaffolding function of NDRG3 (N-Myc downstream-regulated gene 3) promotes specific interaction between KRAS and a deubiquitinating enzyme, USP9X. In KRAS-mutant cancer cells KRAS protein expression, downstream signaling, and cell growth are highly dependent on NDRG3. In conditional KrasG12D knock-in mouse models of pancreatic ductal adenocarcinoma, Ndrg3 depletion abolishes Kras protein expression and suppresses intraepithelial neoplasia formation in pancreas. Mechanistically, KRAS protein binds to the C-terminal serine/threonine-rich region of NDRG3, subsequently going through deubiquitination by USP9X recruited to the complex. This interaction can be disrupted in a dominant-negative manner by a C-terminal NDRG3 fragment that binds KRAS but is defective in USP9X binding, highly suppressing KRAS protein expression and KRAS-driven cell growth. In summary, KRAS-driven cancer development critically depends on the deubiquitination of KRAS protein mediated by USP9X/NDRG3, and KRAS-addicted cancers could be effectively targeted by inhibiting the KRAS-NDRG3 interaction.

Abstract Chimeric antigen receptor T (CAR-T) cells show remarkable efficacy for some hematological malignancies. However, CAR targets that are expressed at high level and selective to tumors are scarce. Several strategies have been proposed to tackle the on-target off-tumor toxicity of CAR-T cells that arise from suboptimal selectivity, but these are complicated, with many involving dual gene expression for specificity. In this study, we show that switchable CAR-T cells with a tumor targeting adaptor can mitigate on-target off-tumor toxicity against a low selectivity tumor antigen that cannot be targeted by conventional CAR-T cells, such as CD40. Our system is composed of anti-cotinine murine CAR-T cells and cotinine-labeled anti-CD40 single chain variable fragments (scFv), with which we show selective tumor killing while sparing CD40-expressing normal cells including macrophages in a mouse model of lymphoma. Simple replacement of the tumor-targeting adaptor with a suicidal drug-conjugated tag may further enhance safety by enabling permanent in vivo depletion of the switchable CAR-T cells when necessary. In summary, our switchable CAR system can control CAR-T cell toxicity while maintaining therapeutic efficacy, thereby expanding the range of CAR targets.

Jing Sang,1,2,* Peng Liu,1,3,* Meixiang Wang,1 Fengkuo Xu,1 Ji Ma,1 Zhigang Wei,1 Xin Ye1 1Department of Oncology, The First Affiliated Hospital of Shandong First Medical University & Shandong Provincial Qianfoshan Hospital, Shandong Lung Cancer Institute, Shandong Key Laboratory of Rheumatic Disease and Translational Medicine, Jinan, People’s Republic of China; 2Department of Pathology, The Affiliated Taian City Central Hospital of Qingdao University, Taian, People’s Republic of China; 3Yuncheng Central Hospital Affiliated to Shanxi Medical University, Yuncheng, People’s Republic of China*These authors contributed equally to this workCorrespondence: Zhigang Wei; Xin Ye, Department of Oncology, The First Affiliated Hospital of Shandong First Medical University & Shandong Provincial Qianfoshan Hospital, Jinan, Shandong Province, 250014, People’s Republic of China, Email weizhigang321321@163.com; yexintaian2020@163.comPurpose: Microwave ablation (MWA) is a minimally invasive technique for treating lung cancer. It can induce immune response; however, its effect on the immune microenvironment in tumor-draining lymph nodes (TdLN) is not well understood. This study aims to identify changes in the immune microenvironment in TdLN following MWA in a Lewis lung cancer (LLC) mouse model.Methods: LLC mouse model was established and followed by MWA. TdLN were collected at various time points, including pre-MWA and days 1, 2, 4, and 8 post-MWA. Flow cytometry was used to determine the frequencies of CD4+ T cells, CD8+ T cells, regulatory T (Treg) cells, natural killer (NK) cells, dendritic cells (DCs) and other immune cells in the TdLN. Certain cytokines were also detected.Results: Compared with pre-MWA, the frequency of CD4+ T cells significantly increased from day 1 to day 8 post-MWA. The frequency of CD8+ T cells decreased significantly on days 2 and 4, but no significant changes occurred on days 1 and 8. Significant decreases in the frequencies of Treg cells and Klrg1+ Treg cells were observed from day 1 to day 4. On days 4 and 8, there was a significant increase in the frequency of NK cells. The frequency of resident cDC2 significantly increased on day 4, whereas CD11b+ migratory cDCs increased on day 1. Additionally, on day 4, a notable rise was observed in the frequency of NK cells secreting IFN-γ, while on day 8, there was a significant increase in the frequency of CD8+ T cells secreting both IFN-γ and TNF-α.Conclusion: MWA of lung cancer can alter the immune microenvironment in the TdLN, triggering immune responses. These changes are particularly evident and intricate within the initial 4 days post-MWA. Treatment combined with MWA within a certain period may significantly enhance anti-tumor immunity.Keywords: microwave ablation, lung cancer, T cells, regulatory T cells, natural killer cells, dendritic cells

Background/Objectives: Occlusion plays a crucial role in the long-term success and functionality of dental restorations. The purpose of this study was to investigate the changes in occlusal contacts upon the cementation of zirconia crowns with different cement spacer settings in computer-aided design and computer-aided manufacturing (CAD-CAM) software (3Shape Dental System version 2.102.1.0). Methods: A master model of a prepared abutment for a crown on the right maxillary first molar was scanned, and 30 sets of sample casts and zirconia crowns were fabricated with varying cement spaces (70 μm and 120 μm). These casts were mounted in maximal intercuspation (MIP) on a semi-adjustable articulator. Pre-cementation adjustments were made to fit the crowns and maintain the existing occlusion. Occlusal records were taken before and after cementation using polyvinyl siloxane impression material. These records were analyzed using a DC light box and image analyzer to measure changes in contact area, intensity, and patterns. Paired sample t-tests were used to compare pre- and post-cementation occlusal contact areas of each sample (α = 0.05). Results: Significant differences in occlusal contact areas were found between pre- and post-cementation in both groups (p < 0.001). The mean post-cementation contact surface area for the 70 μm group was 6281 ± 3310 μm2, compared to 2339 ± 1206 μm2 before cementation. For the 120 μm group, the post-cementation area was 5545 ± 3491 μm2, compared to 2071 ± 909 μm2 before cementation. An increase in contact intensity was also observed after cementation. Conclusions: This study demonstrates that cementation increases occlusal contact surface area and intensity in both cement space groups.

Ginkgo biloba L. (Ginkgoacea) contains an abundance of beneficial compounds and has demonstrated positive clinical effects in the management of metabolic syndrome. Recent studies have emphasized its efficacy against type 2 diabetes mellitus (T2DM), including improvements in diabetic nephropathy and retinopathy. Particularly noteworthy are ginkgolic acid analogs, which have shown potential in combating T2DM by inhibiting protein tyrosine phosphatases (PTPs), facilitating glucose uptake, and influencing signaling pathways. In this study, we isolated six derivatives of ginkgolic acid from the MeOH extract of G. biloba leaves with the guidance of liquid chromatography–mass spectrometry (LC/MS). We determined the chemical structures of these isolated compounds as 2-hydroxy-6-(10′-hydroxypentadec-11′(E)-en-1-yl) benzoic acid (1), 2-hydroxy-6-(11′-hydroxypentadec-9′(E)-en-1-yl) benzoic acid (2), 2-hydroxy-6-tridecylbenzoic acid (3), 2-hydroxy-6-pentadecylbenzoic acid (4), 2-hydroxy-6-(12′-hydroxyheptadec-13′(E)-en-1-yl) benzoic acid (5), and 2-hydroxy-6-(11-hydroxyundecyl) benzoic acid (6) using NMR spectroscopic data and LC/MS analysis. To assess their potential for addressing T2DM, we subjected the isolated compounds (1–6) to tests measuring their inhibitory activity against six PTPs: PTPN11, PTPN2, PTP1B, DUSP9, PTPRS, and PTPN9. Among these compounds, compounds 3–5 displayed enzyme inhibition exceeding 90% against all six PTPs. In conclusion, ginkgolic acid derivatives, acting as PTP inhibitors relevant to insulin resistance, hold promise as potential therapeutic candidates for the prevention and treatment of T2DM.

Purpose. This narrative review aims to provide an overview of the mechanisms of 3D printing, the dental materials relevant to each mechanism, and the possible applications of these materials within different areas of dentistry. Methods. Subtopics within 3D printing technology in dentistry were identified and divided among five reviewers. Electronic searches of the Medline (PubMed) database were performed with the following search keywords: 3D printing, digital light processing, stereolithography, digital dentistry, dental materials, and a combination of the keywords. For this review, only studies or review papers investigating 3D printing technology for dental or medical applications were included. Due to the nature of this review, no formal evidence-based quality assessment was performed, and the search was limited to the English language without further restrictions. Results. A total of 64 articles were included. The significant applications, applied materials, limitations, and future directions of 3D printing technology were reviewed. Subtopics include the chronological evolution of 3D printing technology, the mechanisms of 3D printing technologies along with different printable materials with unique biomechanical properties, and the wide range of applications for 3D printing in dentistry. Conclusions: This review article gives an overview of the history and evolution of 3D printing technology, as well as its associated advantages and disadvantages. Current 3D printing technologies include stereolithography, digital light processing, fused deposition modeling, selective laser sintering/melting, photopolymer jetting, powder binder, and 3D laser bioprinting. The main categories of 3D printing materials are polymers, metals, and ceramics. Despite limitations in printing accuracy and quality, 3D printing technology is now able to offer us a wide variety of potential applications in different fields of dentistry, including prosthodontics, implantology, oral and maxillofacial, orthodontics, endodontics, and periodontics. Understanding the existing spectrum of 3D printing applications in dentistry will serve to further expand its use in the dental field. Three-dimensional printing technology has brought about a paradigm shift in the delivery of clinical care in medicine and dentistry. The clinical use of 3D printing has created versatile applications which streamline our digital workflow. Technological advancements have also paved the way for the integration of new dental materials into dentistry.

ABSTRACT: Ceramide-containing phospholipids improve skin hydration and barrier function and are ideal for use in skin care products. In this study, we evaluated the photoprotective effect of milk phospholipids on the skin condition of UVB-irradiated hairless mice. Skin parameters were assessed following oral administration of milk phospholipids. The UVB irradiation induced photoaging in mice. The animals were divided into 5 groups: a control group (oral administration of saline with no UBV irradiation), UVB group (oral administration of saline with UVB irradiation), and 3 UVB irradiation groups receiving the milk phospholipids at 3 different concentrations of oral administration, 50 mg/kg (ML group), 100 mg/kg (MM group), and 150 mg/kg (MH group), for 8 wk. An increase in skin hydration and transepidermal water loss were improved in the 150 mg/kg of milk phospholipid–administered group. Hematoxylin and eosin staining revealed a decrease in epidermal thickness in the milk phospholipid–administered groups (50, 100, and 150 mg/kg of body weight). In particular, the 100 and 150 mg/kg groups showed significant changes in the area, length, and depth of the wrinkles compared with the UVB group. Moreover, the gene expression of matrix metalloproteins was attenuated, and that of proinflammatory cytokines, especially tumor necrosis factor-α, was significantly reduced in the milk phospholipid–administered groups than in the UVB group. The reduced ceramide and increased sphingosine-1-phosphate levels in the skin tissue due to UVB exposure were restored to levels similar to those of the control group following milk phospholipid administration. These results were confirmed to be due to the downregulation of protein expression of nuclear factor kappa-B (NF-κB) and phosphorylated IκB-α (inhibitor of κB α). Collectively, oral administration of milk phospholipids improves skin health through a synergistic effect on photoprotective activity.

Background/Aims Coenzyme Q10 (CoQ10), is a promising antioxidant; however, low bioavailability owing to lipid-solubility is a limiting factor. We developed water-soluble CoQ10 (CoQ10-W) and compared its effects with conventional lipid-soluble CoQ10 (CoQ10-L) in an experimental model of chronic tacrolimus (Tac) nephropathy. Methods CoQ10-W was developed from a glycyrrhizic-carnitine mixed layer CoQ10 micelle based on acyltransferases. Chronic nephropathy was induced in rats with 28-day Tac treatment; they were concomitantly treated with CoQ10-L or CoQ10-W. CoQ10 level in plasma and kidney were measured using liquid chromatography–mass spectrometry. CoQ10-W and CoQ10-L effects on Tac-induced nephropathy were assessed in terms of renal function, histopathology, oxidative stress, and apoptotic cell death. Their effects on cell viability and reactive oxygen species (ROS) production were assessed in cultured proximal tubular cells, human kidney 2 (HK-2) cells. Results The plasma CoQ10 level was significantly higher in the CoQ10-W group than in the CoQ10-L group. Tac treatment caused renal dysfunction, typical pathologic lesions, and oxidative stress markers. Serum creatinine was restored in the Tac + CoQ10-L or CoQ10-W groups compared with that in the Tac group. CoQ10-W administration reduced oxidative stress and apoptosis markers. Mitochondrial ultrastructure assessment revealed that the addition of CoQ10-L or CoQ10-W with Tac increased mitochondrial size and number than Tac treatment alone. In vitro investigations revealed that both CoQ10-L and CoQ10-W improved cell viability and reduced ROS production in the Tac-induced HK-2 cell injury. Conclusions CoQ10-W has a better therapeutic effect in Tac-induced renal injury than conventional CoQ10-L, possibly associated with improved CoQ10 bioavailability

Several protein tyrosine phosphatases (PTPs), particularly PTPN1, PTPN2, PTPN6, PTPN9, PTPN11, PTPRS, and DUSP9, are involved in insulin resistance. Therefore, these PTPs could be promising targets for the treatment of type 2 diabetes. Our previous studies revealed that PTPN2 and PTPN6 are potential antidiabetic targets. Therefore, the identification of dual-targeting inhibitors of PTPN2 and PTPN6 could be a potential therapeutic strategy for the treatment or prevention of type 2 diabetes. In this study, we demonstrate that methyl syringate inhibits the catalytic activity of PTPN2 and PTPN6 in vitro, indicating that methyl syringate acts as a dual-targeting inhibitor of PTPN2 and PTPN6. Furthermore, methyl syringate treatment significantly increased glucose uptake in mature 3T3-L1 adipocytes. Additionally, methyl syringate markedly enhanced phosphorylation of adenosine monophosphate-activated protein kinase (AMPK) in 3T3L1 adipocytes. Taken together, our results suggest that methyl syringate, a dual-targeting inhibitor of PTPN2 and PTPN6, is a promising therapeutic candidate for the treatment or prevention of type 2 diabetes.

Background/purpose: There is limited literature on the materials of choice and their properties when repairing 3-D printed resin-based restorations. The objective of this in-vitro study is to determine the shear bond strength of various repair materials to 3D printed SLA (stereolithography) resin. Materials and methods: For Group A (control), fifteen cylinders of 3-D printing SLA resin were printed as one unit of a Ø6.8 × 8 mm (diameter and height) cylindrical block with a Ø3 × 5 mm cylindrical block at the center. For the test groups, forty-five specimen cylinders of 3-D printing SLA resin (Ø6.8 × 8 mm) were fabricated and the surfaces were treated with 3 different test materials: Group B: Poly-Methyl Methacrylate (PMMA); Group C: Bis-acrylic composite resin, and Group D: Bis-GMA composite All specimens were tested using an Instron machine at a crosshead speed of 0.5 mm/min. A Shapiro–Wilk test was used to assess normality within the data, then the data was statistically analyzed by a Mann–Whitney test. Results: There were no statistically significant differences between testing groups, except Group A. Group B displayed mixed (87%) and adhesive (13%) failure at the fractured surface. Group C showed both mixed (60%) and adhesive failure at the fractured surface (40%). All Group D showed mixed fracture patterns, partly cohesive fractured surface within the base cylinder area and partly adhesive fractured surface at the bonded interface. Conclusion: No statistically significant differences in the shear bond strength of the different repair materials to 3D printed cylinders were observed. The 3D printed cylinder repaired with Bis-GMA composite demonstrated the most predictability from the fractography analysis.

Background/Aims Coenzyme Q10 (CoQ10) has antioxidant effects and is commercially available and marketed extensively. However, due to its low bioavailability, its effects are still controversial. We developed a water-soluble CoQ10-based micelle formulation (CoQ10-W) and tested it in an experimental model of tacrolimus (TAC)-induced diabetes mellitus (DM). Methods We developed CoQ10-W from a glycyrrhizic-carnitine mixed layer CoQ10 micelle preparation based on acyltransferases. TAC-induced DM rats were treated with either lipid-soluble CoQ10 (CoQ10-L) or CoQ10-W for 4 weeks. Their plasma and pancreatic CoQ10 concentrations were measured using liquid chromatography- tandem mass spectrometry. The therapeutic efficacies of CoQ10-W and CoQ10-L on TAC-induced DM were compared using functional and morphological parameters and their effects on cell viability and reactive oxygen species (ROS) production were also evaluated in cultured rat insulinoma cells. Results The plasma CoQ10 level was significantly increased in the CoQ10-W group compared to that in the CoQ10-L group. Intraperitoneal glucose tolerance tests and glucose-stimulated insulin secretion revealed that CoQ10-W controlled hyperglycemia and restored insulin secretion significantly better than CoQ10-L. The TAC-mediated decrease in pancreatic islet size was significantly attenuated by CoQ10-W but not by CoQ10-L. TAC-induced oxidative stress and apoptosis were significantly more reduced by CoQ10-W than CoQ10-L. Electron microscopy revealed that CoQ10-W restored TAC-induced attenuation in the number of insulin granules and the average mitochondrial area, unlike CoQ10-L. In vitro studies showed that CoQ10-L and CoQ10-W both improved cell viability and reduced ROS production in TAC-treated islet cells to a similar extent. Conclusions CoQ10-W has better therapeutic efficacy than CoQ10-L in TAC-induced DM.

BackgroundOsteoarthritis (OA) is the most common degenerative joint disease and is characterized by breakdown of joint cartilage. Coenzyme Q10 (CoQ10) exerts diverse biological effects on bone and cartilage; observational studies have suggested that CoQ10 may slow OA progression and inflammation. However, any effect of CoQ10 on OA remains unclear. Here, we investigated the therapeutic utility of CoQ10-micelles.MethodsSeven-week-old male Wistar rats were injected with monosodium iodoacetate (MIA) to induce OA. CoQ10-micelles were administered orally to MIA-induced OA rats; celecoxib served as the positive control. Pain, tissue destruction, and inflammation were measured. The expression levels of catabolic and inflammatory cell death markers were assayed in CoQ10-micelle-treated chondrocytes.ResultsOral supplementation with CoQ10-micelles attenuated OA symptoms remarkably, including pain, tissue destruction, and inflammation. The expression levels of the inflammatory cytokines IL-1β, IL-6, and MMP-13, and of the inflammatory cell death markers RIP1, RIP3, and pMLKL in synovial tissues were significantly reduced by CoQ10-micelle supplementation, suggesting that CoQ10-micelles might attenuate the synovitis of OA. CoQ10-micelle addition to cultured OA chondrocytes reduced the expression levels of catabolic and inflammatory cell death markers.ConclusionsCoQ10-micelles might usefully treat OA.

Objectives. Although digital technology has been widely integrated into dental education, there is limited literature investigating the extent of the integration of computer-aided design and computer-aided manufacturing (CAD-CAM) for removable systems in the dental curriculum. The purpose of this study was to assess the current implementation of CAD-CAM complete and partial dentures in predoctoral (PP) and advanced graduate prosthodontic (AGP) education in US dental schools. The study also aimed to identify potential barriers to its implementation in the dental curriculum. Methods. An online survey with 15 questions was created using online survey software. The survey was distributed to the directors of predoctoral prosthodontics in 56 schools and advanced graduate programs of prosthodontics in 52 schools listed in the 2018–19 American Dental Education Association (ADEA) Directory. Results. The percentage of programs (PP and AGP) implementing CAD-CAM complete dentures (CAD-CAM CDs) and CAD-CAM removable partial dentures (CAD-CAM RPDs) in their didactic, preclinical, and clinical curricula was recorded. CAD-CAM CDs are taught in didactic courses in 54.2% of PP and 65.2% of AGP. However, CAD-CAM RPDs are only taught in 37.5% of PP and 47.8% of AGP. Programs are largely limited by a lack of funds, resources, time, and faculty members. Conclusion. While digital technologies have indeed become more prevalent in dental education, many institutions face barriers to implementation. More research must be conducted in order to support the continued incorporation of digital technologies into dental education.

The aim of this clinical study was to investigate the change in occlusal vertical dimension (OVD) with dental casts mounted on a mechanical articulator using an average axis facebow and on a virtual articulator mounted using the Bonwill triangle and the Balkwill angle and compare these groups with OVD change observed clinically in patients. Casts were obtained from each patient (n = 14) and mounted on a semi-adjustable articulator in the facebow preservation group (FPG) and on a virtual articulator using average anatomic values in the average mounting group (AMG). Customized mandibular anterior splints were virtually designed at an OVD increased by 3, 6, and 9 mm. Digital buccal scans were performed with the anterior devices in the participants’ mouths in the intraoral group (IOG), AMG, and FPG at the different OVD increases accordingly. While no statistically significant differences (p > 0.05) were observed in the posterior interocclusal measurements with the incisal guide pin raised by 3 mm and 6 mm among all groups, a 9 mm increase resulted in a significant difference between AMG and IOG. The interocclusal posterior-to-anterior opening ratio observed clinically was 1:1.575. Increases in OVD up to 6 mm on dental casts mounted using average anatomic values performed similarly to the actual intraoral changes.

The regulation of protein tyrosine phosphorylation by protein tyrosine kinases and protein tyrosine phosphatases (PTPs) is involved in intracellular signaling pathways, such as cell proliferation, differentiation, migration, and metabolism. As inhibitors of certain PTPs associated with insulin resistance have been shown to improve insulin sensitivity, the use of inhibitors against PTPN1, PTPN9 or PTPN11 is considered an effective strategy for treating type 2 diabetes. Herein, for the first time, we revealed that linoleic acid (LA) inhibited the catalytic activity of PTPN1, PTPN9 and PTPN11 in vitro, indicating that LA targeted PTPN1, PTPN9, and PTPN11. Additionally, 40 μM LA treatment enhanced glucose uptake through activation of the AMPK and Akt signaling pathways. Taken together, these findings indicate that LA, a multi-targeting inhibitor of PTPN1, PTPN9, and PTPN11, may exert antidiabetic effects to prevent type 2 diabetes.

The accurate diagnosis of individual tooth prognosis has to be determined comprehensively in consideration of the broader treatment plan. The objective of this study was to establish an effective artificial intelligence (AI)-based module for an accurate tooth prognosis decision based on the Harvard School of Dental Medicine (HSDM) comprehensive treatment planning curriculum (CTPC). The tooth prognosis of 2359 teeth from 94 cases was evaluated with 1 to 5 levels (1—Hopeless, 5—Good condition for long term) by two groups (Model-A with 16, and Model-B with 13 examiners) based on 17 clinical determining factors selected from the HSDM-CTPC. Three AI machine-learning methods including gradient boosting classifier, decision tree classifier, and random forest classifier were used to create an algorithm. These three methods were evaluated against the gold standard data determined by consensus of three experienced prosthodontists, and their accuracy was analyzed. The decision tree classifier indicated the highest accuracy at 0.8413 (Model-A) and 0.7523 (Model-B). Accuracy with the gradient boosting classifier and the random forest classifier was 0.6896, 0.6687, and 0.8413, 0.7523, respectively. Overall, the decision tree classifier had the best accuracy among the three methods. The study contributes to the implementation of AI in the decision-making process of tooth prognosis in consideration of the treatment plan.

Fang Li,1 Na Li,1 Jiao Sang,1 Xiude Fan,1 Huan Deng,1 Xiaoge Zhang,1 Qunying Han,1 Yi Lv,2,3 Zhengwen Liu1,3 1Department of Infectious Diseases, First Affiliated Hospital of Xi’an Jiaotong University, Xi’an, Shaanxi, China; 2Department of Hepatobiliary Surgery, First Affiliated Hospital of Xi’an Jiaotong University, Xi’an, Shaanxi, China; 3Institute of Advanced Surgical Technology and Engineering, Xi’an Jiaotong University, Xi’an, Shaanxi, China Background and objective: Upregulated T-cell immunoglobulin and mucin domain containing molecule-3 (Tim-3) in hepatitis B virus (HBV)-specific CD8+ T-cells contributes to CD8+ T-cell exhaustion during chronic HBV infection. The membrane-bound Tim-3 can be cleaved from the cell surface by sheddase, yielding soluble Tim-3 (sTim-3). This study investigated serum sTim-3 levels in patients with chronic HBV infection of various liver diseases. Methods: Serum sTim-3 levels were quantitatively determined in 288 patients with chronic HBV infection of various liver diseases. The sTim-3 levels were analyzed in relation to liver diseases including HBV-related hepatocellular carcinoma (HCC) and overall survival of HCC patients. Results: Serum sTim-3 levels in the patients with chronic HBV infection were significantly elevated compared with healthy controls (P3000 pg/mL being related to shorter overall survival than a level ≤3000 pg/mL (P=0.019). Conclusion: Serum sTim-3 is involved in disease progression and HCC development in chronic HBV infection and its quantitative determination may be potentially used as a marker for monitoring the disease progression and predicting the HCC prognosis in chronic HBV infection. Keywords: hepatitis B virus, soluble Tim-3, hepatocellular carcinoma, risk, survival

Protein tyrosine phosphatases (PTPs), along with protein tyrosine kinases, control signaling pathways involved in cell growth, metabolism, differentiation, proliferation, and survival. Several PTPs, such as PTPN1, PTPN2, PTPN9, PTPN11, PTPRS, and DUSP9, disrupt insulin signaling and trigger type 2 diabetes, indicating that PTPs are promising drug targets for the treatment or prevention of type 2 diabetes. As part of an ongoing study on the discovery of pharmacologically active bioactive natural products, we conducted a phytochemical investigation of African mango (Irvingia gabonensis) using liquid chromatography–mass spectrometry (LC/MS)-based analysis, which led to the isolation of terminalin as a major component from the extract of the seeds of I. gabonensis. The structure of terminalin was characterized by spectroscopic methods, including one-dimensional (1D) and two-dimensional (2D) nuclear magnetic resonance (NMR) and high-resolution (HR) electrospray ionization (ESI) mass spectroscopy. Moreover, terminalin was evaluated for its antidiabetic property; terminalin inhibited the catalytic activity of PTPN1, PTPN9, PTPN11, and PTPRS in vitro and led to a significant increase in glucose uptake in differentiated C2C12 muscle cells, indicating that terminalin exhibits antidiabetic effect through the PTP inhibitory mechanism. These findings suggest that terminalin derived from African mango could be used as a functional food ingredient or pharmaceutical supplement for the prevention of type 2 diabetes.

With the advent of a digital workflow in dentistry, the inter-occlusal articulation of digital models is now possible through various means. The Cadent iTero intraoral scanner uses a buccal scan in maximum intercuspation to record the maxillomandibular relationship. This in-vitro study compares the occlusion derived from conventionally articulated stone casts versus that of digitally articulated quadrant milled models. Thirty sets of stone casts poured from full arch polyvinyl siloxane impressions (Group A) and thirty sets of polyurethane quadrant models milled from digital impressions (Group B) were used for this study. The full arch stone casts were hand-articulated and mounted on semi-adjustable articulators, while the digitally derived models were pre-mounted from the milling center based on the data obtained from the buccal scanning procedure. A T-scan sensor was used to obtain a bite registration from each set of models in both groups. The T-scan data derived from groups A and B were compared to that from the master model to evaluate the reproducibility of the occlusion in the two groups. A statistically significant difference of the contact region surface area was found on #11 of the digitally articulated models compared to the master. An analysis of the force distribution also showed a tendency for a heavier distribution on the more anterior #11 tooth for the digitally articulated models. Within the limitations of this study, the use of a digitally articulated quadrant model system may result in a loss of accuracy, in terms of occlusion, the further anteriorly the tooth to be restored is located. Care must be taken to consider the sources of inaccuracies in the digital workflow to minimize them for a more efficient and effective restorative process.

Qianqian Zhu,1 Na Li,1 Fang Li,1 Zhihua Zhou,1 Jiao Sang,1 Xiaoyan Zeng,2 Qunying Han,1 Yi Lv,3,4 Wenxuan Zhao,1 Zhengwen Liu1,4 1Department of Infectious Diseases, 2Department of Laboratory Medicine, 3Department of Hepatobiliary Surgery, The First Affiliated Hospital of Xi’an Jiaotong University, 4Institute of Advanced Surgical Technology and Engineering, Xi’an Jiaotong University, Xi’an, Shaanxi, People’s Republic of China Background and objective: The treatment of hepatocellular carcinoma (HCC) involves multidisciplinary clinical divisions and patients may be admitted to clinical divisions of different disciplinary nature. Few studies have assessed the potential effect of hospital admissions into different divisions on patient treatment options and survival. This study aimed to analyze this potential effect. Methods: We analyzed data of HCC patients between 2002 and 2011 in a large hospital of northwest China and compared the treatment options and patient outcomes following initial admission into two major clinical disciplinary divisions: internal medicine and surgical. Barcelona Clinic Liver Cancer criteria were used for staging. Results: The study included 2,045 patients. Analysis showed that more patients initially admitted to surgical divisions received curative treatments (resection, transplantation, and local ablation) than those admitted to internal medicine divisions; while more patients initially hospitalized to internal medicine divisions chose supportive care than those admitted to surgical divisions. Stages 0, A, and B patients initially admitted to surgical divisions had higher survival rates compared with those initially admitted to internal medicine divisions (P=0.036, 0.057 and 0.001, respectively). Survival rates of patients who were in stages C and D showed no differences. The survival differences between patients initially admitted to internal medicine and surgical divisions vanished after adjusting for treatment distribution. Conclusion: This study showed that the initial hospitalization divisions may affect the outcome of HCC patients because of different treatment options, suggesting that enforcing multidisciplinary collaboration to optimize the treatment of HCC patients at various stages may improve patient survival. Keywords: hepatocellular carcinoma, treatment option, outcome, multidisciplinary cooperation

The enzyme mimetic activity of nanomaterials has been applied in colorimetric assays and point-of-care diagnostics. Several nanomaterials have been exploited for their peroxidase mimetic activity toward 3,3′,5,5′-tetramethylbenzidine (TMB) in the presence of hydrogen peroxide. However, an efficient nanomaterial for the rapid and strong oxidation of TMB remains a strategic challenge. Therefore, in this study, we developed copper-loaded tin oxide (SnO2-Cu) nanocomposites that rapidly oxidize TMB. These nanocomposites have strong absorption at 650 nm and can be used for highly sensitive colorimetric detection. An environmentally friendly (green), rapid, easy, and cost-effective method was developed for the synthesis of these nanocomposites, which were characterized using ultraviolet-visible, energy-dispersive X-ray, and Fourier-transform infrared spectroscopy, as well as scanning electron microscopy. This is the first green synthesis of SnO2-Cu nanocomposites. Their enzyme mimetic activity, which was first studied here, was found to be strongly dependent on the temperature and pH value of the solution. The synthesized nanocomposites have the advantages of low cost, high stability, and ease of preparation for enzyme mimetic applications. Hence, SnO2-Cu nanocomposites are a promising alternative to peroxidase enzymes in colorimetric point-of-care diagnostics.

Inhibition of the megakaryocyte protein tyrosine phosphatase 2 (PTP-MEG2, also named PTPN9) activity has been shown to be a potential therapeutic strategy for the treatment of type 2 diabetes. Previously, we reported that PTP-MEG2 knockdown enhances adenosine monophosphate activated protein kinase (AMPK) phosphorylation, suggesting that PTP-MEG2 may be a potential antidiabetic target. In this study, we found that phloridzin, isolated from Ulmus davidiana var. japonica, inhibits the catalytic activity of PTP-MEG2 (half-inhibitory concentration, IC50 = 32 ± 1.06 μM) in vitro, indicating that it could be a potential antidiabetic drug candidate. Importantly, phloridzin stimulated glucose uptake by differentiated 3T3-L1 adipocytes and C2C12 muscle cells compared to that by the control cells. Moreover, phloridzin led to the enhanced phosphorylation of AMPK and Akt relevant to increased insulin sensitivity. Importantly, phloridzin attenuated palmitate-induced insulin resistance in C2C12 muscle cells. We also found that phloridzin did not accelerate adipocyte differentiation, suggesting that phloridzin improves insulin sensitivity without significant lipid accumulation. Taken together, our results demonstrate that phloridzin, an inhibitor of PTP-MEG2, stimulates glucose uptake through the activation of both AMPK and Akt signaling pathways. These results strongly suggest that phloridzin could be used as a potential therapeutic candidate for the treatment of type 2 diabetes.

Natural products have continued to offer tremendous opportunities for drug development, as they have long been used in traditional medicinal systems. SHP2 has served as an anticancer target. To identify novel SHP2 inhibitors with potential anticancer activity, we screened a library containing 658 natural products. Polyphyllin D was found to selectively inhibit SHP2 over SHP1, whereas two other identified compounds (echinocystic acid and oleanolic acid) demonstrated dual SHP1 and SHP2 inhibition. In a cell-based assay, polyphyllin D exhibited cytotoxicity in Jurkat cells, an acute lymphoma leukemia cell line, whereas the other two compounds were ineffective. Polyphyllin D also decreased the level of phosphorylated extracellular signal-regulated kinase (p-ERK), a proliferation marker in Jurkat cells. Furthermore, knockdown of protein tyrosine phosphatase (PTP)N6 (SHP1) or PTPN11 (SHP2) decreased p-ERK levels. However, concurrent knockdown of PTPN6 and PTPN11 in Jurkat cells recovered p-ERK levels. These results demonstrated that polyphyllin D has potential anticancer activity, which can be attributed to its selective inhibition of SHP2 over SHP1.

Hydrogen peroxide is a low-reactivity reactive oxygen species (ROS); however, it can easily penetrate cell membranes and produce highly reactive hydroxyl radical species through Fenton’s reaction. Its presence in abnormal amounts can lead to serious diseases in humans. Although the development of a simple, ultrasensitive, and selective method for H2O2 detection is crucial, this remains a strategic challenge. The peroxidase mimetic activity of palladium nanoclusters (PdNCs) has not previously been evaluated. In this study, we developed an ultrasensitive and selective colorimetric detection method for H2O2 using PdNCs. An unprecedented eco-friendly, cost-effective, and facile biological method was developed for the synthesis of PdNCs. This is the first report of the biosynthesis of PdNCs. The synthesized nanoclusters had a significantly narrow size distribution profile and high stability. The nanoclusters were demonstrated to possess a peroxidase mimetic activity that could oxidize peroxidase substrate 3,3′,5,5′-tetramethylbenzidine (TMB). Various interfering substances in serum (100 μM phenylalanine, cysteine, tryptophan, arginine, glucose, urea, Na+, Fe2+, PO43−, Mn+2, Ca2+, Mg2+, Zn2+, NH4+, and K+) were included to evaluate the selectivity of the assay, and oxidation of TMB occurred only in the presence of H2O2. Therefore, PdNCs show an efficient nanozyme for the peroxidase mimetic activity. The assay produced a sufficient signal at the ultralow concentration of 0.0625 µM H2O2. This colorimetric assay provides a real-time, rapid, and easy-to-use platform for the detection of H2O2 for clinical purposes.

Recent developments in the area of nanotechnology have focused on the development of nanomaterials with catalytic activities. The enzyme mimics, nanozymes, work efficiently in extreme pH and temperature conditions, and exhibit resistance to protease digestion, in contrast to enzymes. We developed an environment-friendly, cost-effective, and facile biological method for the synthesis of ZnO-Pd nanosheets. This is the first biosynthesis of ZnO-Pd nanosheets. The synthesized nanosheets were characterized by UV–visible spectroscopy, X-ray diffraction (XRD), scanning electron microscopy, transmission electron microscopy, and energy-dispersive X-ray. The d-spacing (inter-atomic spacing) of the palladium nanoparticles in the ZnO sheets was found to be 0.22 nm, which corresponds to the (111) plane. The XRD pattern revealed that the 2θ values of 21.8°, 33.3°, 47.7°, and 56.2° corresponded with the crystal planes of (100), (002), (112), and (201), respectively. The nanosheets were validated to possess peroxidase mimetic activity, which oxidized the 3,3′,5,5′-tetramethylbenzidine (TMB) substrate in the presence of H2O2. After 20 min of incubation time, the colorless TMB substrate oxidized into a dark-blue-colored one and a strong peak was observed at 650 nm. The initial velocities of Pd-ZnO-catalyzed TMB oxidation by H2O2 were analyzed by Michaelis–Menten and Lineweaver–Burk plots, resulting in 64 × 10−6 M, 8.72 × 10−9 Msec−1, and 8.72 × 10−4 sec−1 of KM, Vmax, and kcat, respectively.

The aim of this study was to evaluate the surface topography and the precision measurements of different intraoral and extraoral digital scanners. A reference model of a maxillary arch with four implant analogs was prepared and scanned by three intraoral and two extraoral scanners. The reference model was scanned fifteen times with each digital scanning system, investigating the surface topography and precision measurements for the same-arch and cross-arch measurements. The data was exported to 3D inspection and mesh-processing software (GOM Inspect, Braunschweig, Germany). Statistical analysis was performed using a one-way Analysis of Variance (ANOVA) with the Tukey method for pairwise comparisons. The effect of parameters on generating the surface topography was analyzed by Univariate Linear Regression Analysis. Of the scanner systems evaluated, iTero (IT) exhibited the most number of triangulation points, followed by Trios 3 Shape (TR) and Straumann Cares (SC). There were no significant differences observed in the surface topography when comparing flat and contoured surfaces, the anterior and posterior position, and interproximal areas. For the precision measurement in the same quadrant, no statistical difference was noted between intra- and extraoral scanners. However, the extraoral scanners showed substantially higher precision measurements for the cross-arch measurement. Surface topography did not correlate to precision. Rather, precision correlated with the scanning mechanism. For a quadrant scanning, both intraoral and extraoral scanners are recommended, but extraoral scanners are recommended for a full-arch scanning.

The introduction of digital dentistry and CAD/CAM technology has redefined treatment concepts in implant dentistry—computer guided implant placement has become routine practice, and CAD/CAM prostheses are now commonplace. These advances in treatment options and modalities has led to a paradigm shift in the workflow of surgical and restorative treatments. This case report presents a customized staged treatment protocol that involves the strategic retention of teeth to serve as transitional abutments, which will support a computer guided implant surgical guide as well as a fixed interim prosthesis. The treatment protocol also describes an integrated digital and conventional workflow for full mouth implant-supported fixed prosthetic rehabilitations to provide improved patient care with more predictable outcomes and fewer complications.

Target identification of biologically active molecules such as natural products, synthetic small molecules, peptides, and oligonucleotides mainly relies on affinity chromatography, activity-based probes, or photoaffinity labeling (PAL). Amongst them, activity-based probes and PAL have offered great advantages in target identification technology due to their ability to form covalent bonds with the corresponding targets. Activity-based probe technology mainly relies on the chemical reactivity of the target proteins, thereby limiting the majority of the biological targets to enzymes or proteins which display reactive residues at the probe-binding site. In general, the probes should bear a reactive moiety such as an epoxide, a Michael acceptor, or a reactive alkyl halide in their structures. On the other hand, photoaffinity probes (PAPs) are composed of a target-specific ligand and a photoactivatable functional group. When bound to the corresponding target proteins and activated with wavelength-specific light, PAPs generate highly reactive chemical species that covalently cross-link proximal amino acid residues. This process is better known as PAL and is widely employed to identify cellular targets of biologically active molecules. This review highlights recent advances in target identification by PAL, with a focus on the structure and chemistry of the photoaffinity probes developed in the recent decade, coupled to the target proteins identified using these probes.

Acetylation is one of the most crucial post-translational modifications that affect protein function. Protein lysine acetylation is catalyzed by acetyltransferases, and acetyl-CoA functions as the source of the acetyl group. Additionally, acetyl-CoA plays critical roles in maintaining the balance between carbohydrate metabolism and fatty acid synthesis. Here, we sought to determine whether lysine acetylation is an important process for adipocyte differentiation. Based on an analysis of the acetylome during adipogenesis, various proteins displaying significant quantitative changes were identified by LC-MS/MS. Of these identified proteins, we focused on malate dehydrogenase 1 (MDH1). The acetylation level of MDH1 was increased up to 6-fold at the late stage of adipogenesis. Moreover, overexpression of MDH1 in 3T3-L1 preadipocytes induced a significant increase in the number of cells undergoing adipogenesis. The introduction of mutations to putative lysine acetylation sites showed a significant loss of the ability of cells to undergo adipogenic differentiation. Furthermore, the acetylation of MDH1 dramatically enhanced its enzymatic activity and subsequently increased the intracellular levels of NADPH. These results clearly suggest that adipogenic differentiation may be regulated by the acetylation of MDH1 and that the acetylation of MDH1 is one of the cross-talk mechanisms between adipogenesis and the intracellular energy level.

Enzyme modified electrodes were fabricated with poly(aniline-co-m-aminophenol). Electrochemical polymerization of aniline and m-aminophenol wasperformed to get the film of copolymer on the surface of gold electrode. Modifiedelectrodes were fabricated by two methods, physical entrapment and covalent cross-linking.In one of the method, gold nanoparticles were loaded into the copolymer film andhorseradish peroxidase (HRP) was immobilized into the Au nanoparticle loaded copolymerfilm through physical entrapment. In the other method, the amino and -OH groups in thecopolymer are utilized to form covalent functionalization with HRP via glutaric dialdehydeas cross-linker/mediator. The conducting copolymer/enzyme modified electrodes preparedby physical entrapment/covalent functionalization of enzyme were tested forelectrocatalytic activities towards sensing of H2O2. Amperometric results indicate thatenzyme modified electrode via physical entrapment possesses better electrocatalyticperformance over covalent functionalized enzyme electrode.

PurposeThis study sought to investigate the influence of phoria adaptation on convergence peak velocity from responses located at different initial vergence positions.MethodsSymmetrical 4° convergence step responses and near dissociated phoria (measured at 40 cm from the subject's midline) were recorded from six subjects with normal binocular vision using an infrared limbus tracking system with a haploscope. Two different sustained fixations (1° and 16° convergent rotation along the subject's midline) were used to study whether phoria had an influence on the peak velocity of convergence responses located at two initial vergence positions (1° or 'far' steps and 12° or 'near' steps).ResultsPhoria was significantly adapted after a sustained fixation task at near (16°) and far (1°) (pConclusionAs a result of sustained fixation, phoria was adapted and the peak velocity of the near and far convergence steps was modified. This study has clinical considerations since prisms, which evoke phoria adaptation, can be prescribed to help alleviate visual discomfort. Future investigations should include a systematic study of how prisms may influence convergence and divergence eye movements for those prescribed with prisms within their spectacles.

Traditionally, metal-ceramics, metal-reinforced acrylics, and--more recently--full-contour or layered zirconia have been the materials of choice for definitive fixed implant-supported rehabilitations. Polymethyl methacrylate (PMMA) is commonly used in implant dentistry for the fabrication of implant-supported interim prostheses and as milled or 3D-printed prototypes. This article describes a novel protocol to prosthetically restore a completely edentulous patient following a digital workflow, with fixed, screw-retained, implant-supported prostheses fabricated from CAD/CAM milled PMMA, with no metal substructure. After a 2-year follow-up in terms of esthetics, phonetics, function, and biologic tissue response, the outcome remains functional and free of mechanical, biomechanical, or biologic complications. The aim of this article is to illustrate the feasibility of using milled PMMA as a viable definitive prosthetic material for the fixed implant rehabilitation of edentulous patients. [ABSTRACT FROM AUTHOR]