Frailty is not an independent risk factor for worse clinical outcomes in lumbar spinal surgery: a prospective cohort study.

PURPOSE: Recently, many studies revealed that frailty affects unfavorably on postoperative outcomes in lumbar spinal diseases. This study aimed to investigate the relationship between frailty and clinical outcomes while identifying risk factors associated with worse clinical outcomes following lumbar spinal surgery. METHODS: From March 2019 to February 2021, we prospectively enrolled eligible patients with degenerative lumbar spinal diseases requiring surgery. Frailty was assessed preoperatively. To identify the impact of frailty on lumbar spinal diseases, clinical outcomes, which were measured with patient-reported outcomes (PROs) and postoperative complications, were compared according to the frailty. PROs were assessed preoperatively and one year postoperatively. In addition, risk factors for preoperative and postoperative worse clinical outcomes were investigated. RESULTS: PROs were constantly lower in the frail group than in the non-frail group before and after surgery, and the change of PROs between before and after surgery and postoperative complications were not different between the groups. In addition, frailty was a persistent risk factor for postoperative worse clinical outcome before and after surgery in lumbar spinal surgery. CONCLUSION: Frailty persistently affects the clinical outcome negatively before and after surgery in lumbar spinal surgery. However, as the change of the clinical outcome is not different between the frail group and the non-frail group, it is difficult to interpret whether the frail patients are vulnerable to the surgery. In conclusion, frailty is not an independent risk factor for worse clinical outcome in lumbar spinal surgery.

Preclinical evaluation of general toxicity and safety pharmacology of a receptor-binding domain-based COVID-19 subunit vaccine in various animal models.

The coronavirus disease 2019 pandemic has resulted in the introduction of several naïve methods of vaccine development, which have been used to prepare novel viral vectors and mRNA-based vaccines. However, reluctance to receive vaccines owing to the uncertainty regarding their safety is prevalent. Therefore, rigorous safety evaluation of vaccines through preclinical toxicity studies is critical to determine the safety profiles of vaccine candidates. This study aimed to evaluate the toxicity profile of HuVac-19, a subunit vaccine of SARS-CoV-2 utilizing the receptor-binding domain as an antigen, in rats, rabbits, and dogs using single- and repeat-dose study designs. Repeat-dose toxicity studies in rats and rabbits showed transient changes in hematological and serum biochemical parameters in the adjuvant and/or vaccine groups; however, these changes were reversed or potentially reversible after the recovery period. Moreover, temporary reversible changes in absolute and relative organ weights were observed in the prostate of rats and the thymus of rabbits. Gross examination of the injection sites in rats and rabbits treated with the adjuvant- and HuVac-19 showed discoloration and foci, whereas histopathological examination showed granulomatous inflammation, inflammatory cell infiltration, and myofiber degeneration/necrosis. This inflammatory response was local, unassociated with other toxicological changes, and resolved. In a pharmacological safety study, no toxicological or physiological changes associated with HuVac-19 administration were observed. In conclusion, HuVac-19 was not associated with any major systemic adverse effects in the general toxicity and safety pharmacology evaluation, demonstrating that HuVac-19 is a vaccine candidate with sufficient capacity to be used in human clinical trials.

A Biodegradable Magnetic Microrobot Based on Gelatin Methacrylate for Precise Delivery of Stem Cells with Mass Production Capability.

A great deal of research has focused on small-scale robots for biomedical applications and minimally invasive delivery of therapeutics (e.g., cells, drugs, and genes) to a target area. Conventional fabrication methods, such as two-photon polymerization, can be used to build sophisticated micro- and nanorobots, but the long fabrication cycle for a single microrobot has limited its practical use. This study proposes a biodegradable spherical gelatin methacrylate (GelMA) microrobot for mass production in a microfluidic channel. The proposed microrobot is fabricated in a flow-focusing droplet generator by shearing a mixture of GelMA, photoinitiator, and superparamagnetic iron oxide nanoparticles (SPIONs) with a mixture of oil and surfactant. Human nasal turbinate stem cells (hNTSCs) are loaded on the GelMA microrobot, and the hNTSC-loaded microrobot shows precise rolling motion in response to an external rotating magnetic field. The microrobot is enzymatically degraded by collagenase, and released hNTSCs are proliferated and differentiated into neuronal cells. In addition, the feasibility of the GelMA microrobot as a cell therapeutic delivery system is investigated by measuring electrophysiological activity on a multielectrode array. Such a versatile and fully biodegradable microrobot has the potential for targeted stem cell delivery, proliferation, and differentiation for stem cell-based therapy.

Chrysophanol-Induced Autophagy Disrupts Apoptosis via the PI3K/Akt/mTOR Pathway in Oral Squamous Cell Carcinoma Cells.

Background and Objectives: Natural products are necessary sources for drug discovery and have contributed to cancer chemotherapy over the past few decades. Furthermore, substances derived from plants have fewer side effects. Chrysophanol is an anthraquinone derivative that is isolated from rhubarb. Although the anticancer effect of chrysophanol on several cancer cells has been reported, studies on the antitumor effect of chrysophanol on oral squamous-cell carcinoma (OSCC) cells have yet to be elucidated. Therefore, in this study, we investigated the anticancer effect of chrysophanol on OSCC cells (CAL-27 and Ca9-22) via apoptosis and autophagy, among the cell death pathways. Results: It was found that chrysophanol inhibited the growth and viability of CAL-27 and Ca9-22 and induced apoptosis through the intrinsic pathway. It was also found that chrysophanol activates autophagy-related factors (ATG5, beclin-1, and P62/SQSTM1) and LC3B conversion. That is, chrysophanol activated both apoptosis and autophagy. Here, we focused on the roles of chrysophanol-induced apoptosis and the autophagy pathway. When the autophagy inhibitor 3-MA and PI3K/Akt inhibitor were used to inhibit the autophagy induced by chrysophanol, it was confirmed that the rate of apoptosis significantly increased. Therefore, we confirmed that chrysophanol induces apoptosis and autophagy at the same time, and the induced autophagy plays a role in interfering with apoptosis processes. Conclusions: Therefore, the potential of chrysophanol as an excellent anticancer agent in OSCC was confirmed via this study. Furthermore, the combined treatment of drugs that can inhibit chrysophanol-induced autophagy is expected to have a tremendous synergistic effect in overcoming oral cancer.

New grading system for the clinical evaluation of patients with spinal vascular lesions.

PURPOSE: Neurointerventional approaches have improved myelopathy in patients with spinal vascular lesions by providing effective management, particularly when surgical approaches are difficult. However, there have been challenges in describing and comparing recovery status during the post-treatment period. METHODS: We evaluated 43 patients with venous congestive myelopathy (VCM) using Aminoff-Logue Disability Scale for gait (AL-G) and micturition (AL-M) scores. These results were compared with our new PSMS grading system that evaluates four categories (grades 0-3): pain, sensory symptoms, motor deficit, and sphincter change. Simple linear regression was used to identify the association or trend among the scales. We also calculated an overall area under the receiver operating characteristic curve to compare the predictive ability of the PSMS system with that of the previous grading system (AL-G and AL-M). RESULTS: Compared with other grading system, the PSMS system was more sensitively correlated with patient status and the results were easy to compare with previous clinical statuses during follow-up. The PSMS system also measured pain, which is commonly associated with spinal dural arteriovenous fistula and not precisely evaluated by other grading system. CONCLUSIONS: The new PSMS grading system for patients with VCM correlated well with the previously used systems and included pain evaluation. This new grading system is an easy tool for the evaluation and comparison of outcomes before and after endovascular treatment.

Blocking ß1/ß2-Adrenergic Signaling Reduces Dietary Fat Absorption by Suppressing Expression of Pancreatic Lipase in High Fat-Fed Mice.

We investigated whether ß-adrenergic antagonists attenuates dietary fat absorption through the regulation of pancreatic lipase (PNLIP) expression in pancreatic acinar cells in the context of high fat diet feeding. Male six-week-old C57BL/6 mice were assigned into an ad libitum fed control diet (CON) and a high fat diet (HIGH). Within each diet group, subgroups of mice were treated with vehicle (VEH) or propranolol, a ß-adrenergic antagonist (BB). Over 12 weeks, body weight gain observed in HIGHVEH was mitigated in HIGHBB (+103% vs. +72%). Increase in fecal fat amount observed in HIGHVEH was further increased in HIGHBB. Increase in PNLIP expressions observed in HIGHVEH pancreatic tissues was abolished in HIGHBB. PNLIP expression in mouse primary pancreatic acinar cells and 266-6 cell lines increased with isoproterenol treatment, which was blocked by propranolol. Isoproterenol increased PNLIP expression in a cAMP/protein kinase A/ cyclic AMP response element binding protein (CREB)-dependent manner. CREB directly bound to the CRE on the mouse PNLIP promoter and transactivated PNLIP expression. These results suggest that sympathetic activation increases dietary fat absorption through the upregulation of PNLIP expression and that a ß-adrenergic antagonist attenuates obesity development partly through the downregulation of PNLIP expression and inhibition of dietary fat absorption in the context of high fat diet feeding.

Subjective Study on Pediatric Emergency Department Nurses' Perceptions of Urgency Using Q Methodology.

PURPOSE: In the emergency department (ED), triage significantly impacts patient safety. Therefore, triage nurses must make decisions accurately and timeously. This study aims to investigate how South Korean pediatric emergency nurses perceive urgency and classify severity using the Q methodology, which examines individuals' subjectivity. METHODS: We collected 84 statements from a Q population based on a literature review and interviews, and selected 33 Q samples. The P samples included 30 pediatric emergency nurses at a Seoul tertiary-care hospital. The principal component factor analysis method was employed to analyze data using the PC-QUANL program. RESULTS: Four urgency perception types were identified among pediatric ED nurses. Type 1: "Experiential coping"; Type 2: "Careful reasoning"; Type 3: "Patient-centered thinking"; and Type 4: "Intuitive prediction." These types appear to be an integrated process of knowledge and clinical experience that considers children's characteristics and developmental stages. CONCLUSIONS: This study may serve as a basis for future education to improve pediatric ED nurses' urgency judgment and severity classification skills.

Is it necessary to use a cervical brace after single- or double-level ACDF?

Although anterior cervical discectomy and fusion (ACDF) is one of the most frequently performed spinal surgeries, there is no consensus regarding the necessity of prescribing a cervical brace after surgery. This study aimed to investigate any difference in radiological and clinical outcomes when wearing or not wearing cervical braces after single- or double-level ACDF. We examined 2 cohorts of patients who underwent single- or double-level ACDF surgery with and without a cervical brace: patients who underwent ACDF between March 2018 and December 2019 received a cervical brace, while patients who underwent ACDF between January 2020 and May 2021 did not. Each patient was evaluated radiologically and functionally using plain X-ray, modified Japanese Orthopedic Association score, and visual analog scale for neck and arm until 12 months after surgery. Fusion rate, subsidence, and postoperative complications were also evaluated. Eighty-three patients were included in the analysis: 38 were braced and 45 were not. The demographic characteristics and baseline outcome measures of both groups were similar. There was no statistically significant difference in any of the clinical measures at baseline. The modified Japanese Orthopedic Association score and visual analog scale for neck and arm were similar in both groups at all time intervals and showed statistically significant improvement when compared with preoperative scores. In addition, fusion rate, subsidence, and postoperative complications were similar in both groups. Our results suggest that the use of cervical braces does not improve the clinical outcomes of individuals undergoing single- or double-level ACDF.

The Efficacy of Cervical Pedicle Screw Is Enhanced When Used With 5.5-mm Rods for Metastatic Cervical Spinal Tumor Surgery.

OBJECTIVE: The cervical spine presents challenges in treating metastatic cervical spinal tumors (MCSTs). Although the efficacy of cervical pedicle screw placement (CPS) has been well established, its use in combination with 5.5-mm rods for MCST has not been reported. This study aimed to evaluate the efficacy of CPS combined with 5.5-mm rods in treating MCST and compare it with that of CPS combined with traditional 3.5-mm rods. METHODS: This retrospective study analyzed 58 patients with MCST who underwent posterior cervical spinal fusion surgery by a single surgeon between March 2012 and December 2022. Data included demographics, surgical details, imaging results, numerical rating scale score for neck pain, Eastern Cooperative Oncology Group performance status, and Spine Oncology Study Group Outcomes Questionnaire responses. RESULTS: Preoperative Spinal Instability Neoplastic Scores were significantly higher in the 5.5-mm rod group. Greater kyphotic changes in the index vertebra were observed in the 3.5-mm rod group. Neck pain reduction was significantly better in the 5.5-mm rod group. CONCLUSION: CPS with 5.5-mm rods provides superior biomechanical stability and effectively resists forward bending momentum in posterior MCST fusion surgery. These findings support the use of 5.5-mm rods to enhance surgical outcomes.

Quantitative Analysis of the Effect of Stereotactic Radiosurgery for Postoperative Residual Cervical Dumbbell Tumors: A Multicenter Retrospective Cohort Study.

OBJECTIVE: Stereotactic radiosurgery (SRS) has been performed for spinal tumors. However, the quantitative effect of SRS on postoperative residual cervical dumbbell tumors remains unknown. This study aimed to quantitatively evaluate the efficacy of SRS for treating postoperative residual cervical dumbbell tumors. METHODS: We retrospectively reviewed cases of postoperative residual cervical dumbbell tumors from 1995 to 2020 in 2 tertiary institutions. Residual tumors underwent SRS (SRS group) or were observed with clinical and magnetic resonance imaging (MRI) follow-up (observation group). Tumor regrowth rates were compared between the SRS and observation groups. Additionally, risk factors for tumor regrowth were analyzed. RESULTS: A total of 28 cervical dumbbell tumors were incompletely resected. Eight patients were in the SRS group, and 20 in the observation group. The mean regrowth rate was not significantly lower (p = 0.784) in the SRS group (0.18 ± 0.29 mm/mo) than in the observation group (0.33 ± 0.40 mm/mo). In the multivariable Cox regression analysis, SRS was not a significant variable (hazard ratio [HR], 0.57; 95% confidence interval [CI], 0.18-1.79; p = 0.336). CONCLUSION: SRS did not significantly decrease the tumor regrowth rate in our study. We believe that achieving maximal resection during the initial operation is more important than postoperative adjuvant SRS.

Comparison of the Efficacy of Romosozumab and Teriparatide for the Management of Osteoporotic Vertebral Compression Fractures.

OBJECTIVE: Romosozumab is increasingly employed to manage osteoporosis. However, no studies have analyzed its effects on recent osteoporotic vertebral compression fractures (OVCFs). Therefore, this study aimed to evaluate the efficacy of romosozumab compared with teriparatide in managing OVCFs. METHODS: The electronic medical records of postmenopausal patients with recent OVCFs who were administered romosozumab or teriparatide for one year from March 2018 to August 2022 were retrospectively reviewed. We compared the 2 groups for demographics, radiological outcomes (compression ratio, Cobb angle, and bone mineral density [BMD]), and clinical outcomes (Numerical Rating Scale [NRS] for back pain). RESULTS: Fifty-five patients with OVCFs, 32 patients treated with romosozumab and 23 with teriparatide, were included in this study. The change of BMD (g/cm2) values was significantly higher (p = 0.016) in the romosozumab (0.04 ± 0.06) than in the teriparatide group (0.00 ± 0.08) in the femur total. Furthermore, in subgroup analysis, the change of BMD (g/cm2) values in the lumbar spine was significantly higher (p = 0.016) in the romosozumab (0.12 ± 0.06) than in the teriparatide group (0.07 ± 0.06) in the lumbar spine. The decrease in NRS was significantly higher (p = 0.013) in the romosozumab (6.6 ± 2.0) than in the teriparatide group (5.5 ± 2.1). However, there was no significant difference in radiologic outcomes between the 2 groups. CONCLUSION: Our findings suggest that romosozumab may be more effective than teriparatide in treating OVCFs in postmenopausal females, particularly in improving BMD and reducing back pain as measured by NRS.

Surgical strategy for metastatic spinal tumor patients with surgically challenging situation.

The incidence of spinal metastasis is increasing as cancer patients live longer owing to the improvement of cancer treatments. However, traditional surgery (TS) which fixates at least 2 levels above and 2 levels below the affected vertebrae is sometimes difficult to perform as it is burdensome to the patients. In this article, we introduce our experience and strategy in treating spinal metastasis, focusing particularly on challenging cases. We retrospectively reviewed the data of 110 patients who underwent spinal surgery for metastatic spinal tumors from April 2018 to March 2020. Among them, 5 patients who received anterior approach surgery were excluded. The remaining 105 patients were enrolled. In addition to TS, we also performed cervical pedicle screw, cervicothoracic junction fixation, thoracolumbar short fixation, and decompression surgery, depending on the characteristics of the tumor. The overall survival was analyzed, and the local tumor control rate was evaluated using magnetic resonance imaging. Perioperative clinical characteristics including Spine Oncology Study Group Outcomes Questionnaire, visual analog scale, Eastern Cooperative Oncology Group performance score, and Karnofsky Performance Score were also investigated. The overall survival rate was 57.9% at 1 year, and the local tumor control rate was 81.1% after surgery. There was a statistically significant difference according to the type of the tumor in the survival analysis: the overall survival rates were 72.7% for favorable tumors and 48.6% for unfavorable tumors at 12 months after surgery (P = .04). Spine Oncology Study Group Outcomes Questionnaire, visual analog scale, Eastern Cooperative Oncology Group performance score, and Karnofsky Performance Score was improved after surgery. All surgical methods, including TS, cervical pedicle screw, cervicothoracic junction fixation, thoracolumbar short fixation, and decompression surgery, showed good clinical and radiological outcomes. Optimized surgical methods show similarly good clinical outcomes in managing spinal metastasis as TS.

Undercarboxylated, But Not Carboxylated, Osteocalcin Suppresses TNF-α-Induced Inflammatory Signaling Pathway in Myoblasts.

Undercarboxylated osteocalcin (ucOCN) has been considered to be an important endocrine factor, especially to regulate bone and energy metabolism. Even with the mounting evidence showing the consistent inverse correlation of ucOCN levels in chronic inflammatory diseases, however, the mechanism underlying the involvement of ucOCN in the muscular inflammation has not been fully understood. In the present study, we explored 1) the endocrine role of ucOCN in the regulation of inflammation in C2C12 myoblasts and primary myoblasts and the underlying intracellular signaling mechanisms, and 2) whether G protein-coupled receptor family C group 6 member A (GPRC6A) is the ucOCN-sensing receptor associated with the ucOCN-mediated anti-inflammatory signaling pathway in myoblasts. ucOCN suppressed the tumor necrosis factor-α (TNF-α)-induced expressions of major inflammatory cytokines, including interleukin-1ß (IL-1ß) and inhibited the TNF-α-stimulated activities of transcription factors, including NF-κB, in C2C12 and primary myoblasts. Both knockdown and knockout of GPRC6A, by using siRNA or a CRISPR/CAS9 system, respectively, did not reverse the effect of ucOCN on IL-1ß expression in myoblasts. Interestingly, TNF-α-induced IL-1ß expression was inhibited by knockdown or deletion of GPRC6A itself, regardless of the ucOCN treatment. ucOCN was rapidly internalized into the cytoplasmic region via caveolae-mediated endocytosis, suggesting the presence of new target proteins in the cell membrane and/or in the cytoplasm for interaction with ucOCN in myoblasts. Taken together, these findings indicate that ucOCN suppresses the TNF-α-induced inflammatory signaling pathway in myoblasts. GPRC6A is not a sensing receptor associated with the ucOCN-mediated anti-inflammatory signaling pathway in myoblasts.

Fabrication and Characterization of Graphene Oxide-Coated Plate for Efficient Culture of Stem Cells.

BACKGROUND: For stem cell applications in regenerative medicine, it is very important to produce high-quality stem cells in large quantities in a short time period. Recently, many studies have shown big potential of graphene oxide as a biocompatible substance to enhance cell growth. We investigated if graphene oxide-coated culture plate can promote production efficiency of stem cells. METHODS: Three types of graphene oxide were used for this study. They are highly concentrated graphene oxide solution, single-layer graphene oxide solution, and ultra-highly concentrated single-layer graphene oxide solution with different single-layer ratios, and coated on cell culture plates using a spray coating method. Physiochemical and biological properties of graphene oxide-coated surface were analyzed by atomic force microscope (AFM), scanning electron microscope (SEM), cell counting kit, a live/dead assay kit, and confocal imaging. RESULTS: Graphene oxide was evenly coated on cell culture plates with a roughness of 6.4 ~ 38.2 nm, as measured by SEM and AFM. Young's Modulus value was up to 115.1 GPa, confirming that graphene oxide was strongly glued to the surface. The ex vivo stem cell expansion efficiency was enhanced as bone marrow-derived stem cell doubling time on the graphene oxide decreased compared to the control (no graphene oxide coating), from 64 to 58 h, and the growth rate increased up to 145%. We also observed faster attachment and higher affinity of stem cells to the graphene oxide compared to control by confocal microscope. CONCLUSION: This study demonstrated that graphene oxide dramatically enhanced the ex vivo expansion efficiency of stem cells. Spray coating enabled an ultra-thin coating of graphene oxide on cell culture plates. The results supported that utilization of graphene oxide on culture plates can be a promising mean for mass production of stem cells for commercial applications.

Antimicrobial photodynamic therapy efficacy against specific pathogenic periodontitis bacterial species.

BACKGROUND: To determine the safety and efficacy of antimicrobial photodynamic therapy (aPDT) combination of 0.33 mM Toluidine Blue O (TBO) with 60 mW/cm2 LED irradiation for 5 min that we had established, this study investigated the cytotoxic effect of aPDT combination on mammalian oral cells (gingival fibroblast and periodontal ligament cells) and compared the antimicrobial efficacy of antibiotics (the combination of amoxicillin (AMX) and metronidazole (MTZ)) against representative periodontitis pathogenic bacteria (Porphyromonas gingivalis, Fusobacterium nucleatum, and Aggregatibacter actinomycetemcomitans) versus our aPDT combination. RESULT: aPDT combination did not show any detectable effect on the viability of Streptococcus sanguinis or Streptococcus mitis, the most common resident species in the oral flora. However, it significantly reduced CFU values of P. gingivalis, F. nucleatum, and A. actinomycetemcomitans. The cytotoxicity of the present aPDT combination to mammalian oral cells was comparable to that of standard antiseptics used in oral cavity. In antimicrobial efficacy test, the present aPDT combination showed equivalent bactericidal rate compared to the combination of AMX + MTZ, the most widely used antibiotics in the periodontitis treatment. The bactericidal ability of the AMX + MTZ combination was effective against all five bacteria tested regardless of the bacterial species, whereas the bactericidal ability of the aPDT combination was effective only against P. gingivalis, F. nucleatum, and A. actinomycetemcomitans, the representative periodontitis pathogenic bacterial species. CONCLUSION: The present study demonstrated the safety and efficacy of the present aPDT combination in periodontitis treatment. TBO-mediated aPDT with LED irradiation has the potential to serve as a safe single or adjunctive antimicrobial procedure for nonsurgical periodontal treatment without damaging adjacent normal oral tissue or resident flora.

Monoolein cubic phases containing cinnamic acid, poly(ethyleneimine) and gold nanoparticle and their UV- and NIR-responsive release property.

UV and NIR-responsive monoolein cubic phase was prepared by including poly(ethyleneimine) (PEI)/cinnamic acid (CA) conjugate and gold nanoparticle (GNP) within its structure. UV irradiation elevated significantly the release % of Auramine O loaded in cubic phase containing PEI/CA, possibly because of the trans-to-cis isomerization of CA. NIR irradiation also increased significantly the release % of FITC-dextran loaded in cubic phase containing PEI/CA. The release % of the dye loaded in cubic phase containing PEI/CA and GNP was elevated more markedly by NIR irradiation, possibly due to the phase transition of cubic phase and the disassembling of PEI/CA assembly.

Undercarboxylated osteocalcin downregulates pancreatic lipase expression in an ATF4-dependent manner in pancreatic acinar cells.

Osteocalcin is an osteoblast-specific secreted protein that has been associated with endocrine roles in multiple aspects of energy metabolism. We examined whether undercarboxylated osteocalcin (ucOC) downregulates pancreatic lipase (PNLIP) expression in pancreatic acinar cells and then identified the downstream signaling pathway involved. We previously demonstrated that ß adrenergic blockade attenuates body weight/fat mass gain in high-fat diet-fed mice and that this effect is associated with decreased PNLIP expression in pancreatic acinar cells. In the present study, we first confirmed that the serum ucOC level is inversely correlated with PNLIP expression, i.e., mice exhibiting high serum levels of ucOC showed low PNLIP levels in the pancreas. In in vitro experiments using primary pancreatic acinar and 266-6 cells, ucOC downregulated PNLIP expression. cAMP/PKA signaling inhibitors significantly reversed ucOC-induced downregulation of PNLIP expression. ucOC promoted the phosphorylation of cAMP response element-binding protein 2 (ATF4). Overexpression of ATF4 significantly suppressed PNLIP expression. Knockdown of ATF4 by siRNA reversed the ucOC-induced downregulation of PNLIP expression. A luciferase reporter assay showed that ucOC suppressed PNLIP promoter transactivation. Chromatin immunoprecipitation and a luciferase reporter assay demonstrated that ATF4 directly bound to the CRE on the mouse PNLIP promoter and suppressed PNLIP transactivation. Knockdown of G-protein coupled receptor 6A (Gprc6a), a candidate receptor for mediating the response to ucOC in the bone-pancreas endocrine loop, by siRNA reversed the downregulating effect of ucOC on PNLIP expression. Taken together, ucOC downregulates pancreatic lipase expression in a cAMP/protein kinase A/ATF4-dependent manner. Gprc6a is a potential osteocalcin-sensing receptor that regulates PNLIP expression in pancreatic acinar cells.

Cudraxanthone D Regulates Epithelial-Mesenchymal Transition by Autophagy Inhibition in Oral Squamous Cell Carcinoma Cell Lines.

Cudraxanthone D (CD), derived from the root bark of Cudrania tricuspidata, is a natural xanthone compound. However, the biological activity of CD in terms of human metabolism has been barely reported to date. Autophagy is known as a self-degradation process related to cancer cell viability and metastasis. Herein, we investigated the effects of CD on human oral squamous cell carcinoma (OSCC) metastatic related cell phenotype. We confirmed that CD effectively decreased proliferation and viability in a time- and dose-dependent manner in human OSCC cells. In addition, OSCC cell migration, invasion, and EMT were inhibited by CD. To further determine the underlying mechanism of CD's inhibition of cell metastatic potential, we established the relationship between EMT and autophagy in OSCC cells. Thus, our findings indicated that CD inhibited the potential metastatic abilities of OSCC cells by attenuating autophagy.

Stent-Assisted Coil Embolization of MCA Bifurcation Aneurysms at a Hypoplastic M1 Branch by Use of Hook Technique.

BACKGROUND: We describe the Hook technique enabling coil embolization in unfavorable M1 bifurcation aneurysms and analyze the morphologic variations in M1 bifurcation to evaluate how often such aneurysms can be seen. METHODS: Among 42 MCA aneurysms treated by stent-assisted coil embolization, aneurysms arising at the acute-angled hypoplastic M1 branch (n = 14) were treated by the Hook technique, in which a short stent was deployed only to protect the aneurysm neck after microcatheter selection of the hypoplastic M2, followed by subsequent coiling of the aneurysm. Outcome was evaluated, including Raymond classification, coil packing density, final modified Rankin Scale (mRS), and recurrence. Separately, 100 middle cerebral artery (MCA) bifurcation aneurysms were analyzed to assess the proportion of such unfavorable aneurysms. RESULTS: Procedural success of the Hook technique was obtained in 13 of 14 patients (93%). A mean packing density of 30% was achieved. Magnetic resonance angiographic follow-up at a median 4 months (range, 1-26 months) showed complete occlusion in 11 patients and residual neck filling in 3 patients. There was no clinical event (mRS = 0) over a median 17 months (range, 2-26 months) of clinical follow-up. One patient had a thrombotic occlusion during the procedure, which was resolved after tirofiban infusion, without evidence of an infarct or deficit. Of the 100 MCA bifurcation aneurysms, aneurysm arising in asymmetric hypoplastic M2 division was the most common type (48%). CONCLUSIONS: The Hook technique enabled stent-assisted coiling of M1 bifurcation aneurysm with extension along the asymmetric hypoplastic M2 division and also securing the M2 branch.

Non-Invasive Photodynamic Therapy against -Periodontitis-causing Bacteria.

Periodontitis is initiated by causative bacteria in the gingival sulcus. However, as the lesion is often deep and out of circulation system and biofilm is frequently formed on the bacteria cluster, use of antibacterial agents has been limited and the invasive method such as curettage is thought as an only treatment. Here we designed non-invasive photodynamic therapy (PDT), with the ointment which leads a photosensitizer deliverable into gingival sulcus. We assessed whether 650 nm light-emitting-diode (LED) penetrates the 3-mm soft tissue and effectively activates a photosensitizer toluidine-blue-O (TBO) through the thickness to remove Porphyromonas gingivalis and Fusobacterium nucleatum species. The oral ointment formulation was optimized to efficiently deliver the photosensitizer into gingival sulcus and its efficacy of PDT was evaluated in in vitro and in vivo models. Four weeks of TBO-formulation mediated-PDT treatment significantly attenuated periodontitis-induced alveolar bone loss and inflammatory cytokines production in rats. These results confirm that a 650 nm LED indeed penetrates the gingiva and activates our TBO formulation which is sufficiently delivered to, and retained within, the gingival sulcus; thus, it effectively kills the bacteria that reside around the gingival sulcus. Collectively, TBO-mediated PDT using LED irradiation has potential as a safe adjunctive procedure for periodontitis treatment.