骨－歯根膜線維の複合組織形成による三次元的な歯周組織再生技術の開発

Periodontal tissue is a distinctive tissue structure composed three-dimensionally of cementum, periodontal ligament (PDL) and alveolar bone. Severe periodontal diseases cause fundamental problems for oral function and general health, and conventional dental treatments are insufficient for healing to healthy periodontal tissue. Cell sheet technology has been used in many tissue regenerations, including periodontal tissue, to transplant appropriate stem/progenitor cells for tissue regeneration of a target site as a uniform tissue. However, it is still difficult to construct a three-dimensional structure of complex tissue composed of multiple types of cells, and the transplantation of a single cell sheet cannot sufficiently regenerate a large-scale tissue injury. Here, we fabricated a three-dimensional complex cell sheet composed of a bone-ligament structure by layering PDL cells and osteoblast-like cells on a temperature responsive culture dish. Following ectopic and orthotopic transplantation, only the complex cell sheet group was demonstrated to anatomically regenerate the bone-ligament structure along with the functional connection of PDL-like fibers to the tooth root and alveolar bone. This study represents successful three-dimensional tissue regeneration of a large-scale tissue injury using a bioengineered tissue designed to simulate the anatomical structure

Dentin-pulp regeneration by 3D layered cell sheet

The dentin-pulp complex is a unique structure in teeth that contains both hard and soft tissues. Generally, deep caries and trauma cause damage to the dentin-pulp complex, and if left untreated, this damage will progress to irreversible pulpitis. The aim of this study was to fabricate a layered cell sheet composed of rat dental pulp (DP) cells and odontogenic differentiation of pulp (OD) cells and to investigate the ability to regenerate the dentin-pulp complex in a scaffold tooth. We fabricated two single cell sheets composed of DP cells (DP cell sheet) or OD cells (OD cell sheet) and a layered cell sheet made by layering both cells. The characteristics of the fabricated cell sheets were analyzed using light microscopy, scanning electron microscopy (SEM), hematoxylin-eosin (HE) staining, and immunohistochemistry (IHC). Furthermore, the cell sheets were transplanted into the subrenal capsule of immunocompromised mice for 8 weeks. Following this, the regenerative capacity to form dentin-like tissue was evaluated using micro-computed tomography (Micro-CT), HE staining, and IHC. The findings of SEM and IHC confirmed that layered cell sheets fabricated by stacking OD cells and DP cells maintained their cytological characteristics. Micro-CT of layered cell sheet transplants revealed a mineralized capping of the access cavity in the crown area, similar to that of natural dentin. In contrast, the OD cell sheet group demonstrated the formation of irregular fragments of mineralized tissue in the pulp cavity, and the DP cell sheet did not develop any hard tissue. Moreover, bone volume/tissue volume (BV/TV) showed a significant increase in hard tissue formation in the layered cell sheet group compared to that in the single cell sheet group (p<0.05). HE staining also showed a combination of soft and hard tissue formation in the layered cell sheet group. Furthermore, IHC confirmed that the dentin-like tissue generated from the layered cell sheet expressed characteristic markers of dentin but not bone equivalent to that of a natural tooth. In conclusion, this study demonstrates the feasibility of regenerating dentin-pulp complex using a bioengineered tissue designed to simulate the anatomical structure

Macrophage deletion of Noc4l triggers endosomal TLR4/TRIF signal and leads to insulin resistance

In obesity, macrophages drive a low-grade systemic inflammation (LSI) and insulin resistance (IR). The ribosome biosynthesis protein NOC4 (NOC4) mediates 40 S ribosomal subunits synthesis in yeast. Hereby, we reported an unexpected location and function of NOC4L, which was preferentially expressed in human and mouse macrophages. NOC4L was decreased in both obese human and mice. The macrophage-specific deletion of Noc4l in mice displayed IR and LSI. Conversely, Noc4l overexpression by lentivirus treatment and transgenic mouse model improved glucose metabolism in mice. Importantly, we found that Noc4l can interact with TLR4 to inhibit its endocytosis and block the TRIF pathway, thereafter ameliorated LSI and IR in mice.Macrophage inflammation promotes insulin resistance during diet-induced obesity. Here the authors show that macrophage NOC4L is decreased in humans and mice with obesity, that macrophage NOC4L deficiency aggravated high-fat diet induced inflammation and insulin resistance, and that NOC4L interacts with toll-like receptor 4, to inhibit endocytosis, and thus blocks TLF4/TRIF inflammatory signaling

Chemical Constituents of the Ethyl Acetate Extract from Diaphragma juglandis Fructus and Their Inhibitory Activity on Nitric Oxide Production In Vitro

Diaphragma juglandis fructus contains various bioactive constituents. Fourteen compounds were isolated from Diaphragma juglandis fructus by preparative high performance liquid chromatography (pre-HPLC) and high-speed counter-current chromatography (HSCCC). Their structures were identified by nuclear magnetic resonance (NMR) and electrospray ionization mass spectrometry (ESI-MS). Compounds (+)-dehydrovomifoliol (12), (6R,9R)-9-hydroxymegastigman-4-en-3-one (13) and (6R,9S)-9-hydroxymegastigman-4-en-3-one (14) are found from Juglans regia L. for the first time. Compounds dihydrophaseic acid (2), blumenol B (3) and (4S)-4-hydroxy-1-tetralone (11) are isolated from Diaphragma juglandis fructus for the first time. The anti-inflammatory effects of isolated compounds were evaluated by an in vitro model of lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. Compounds gallic acid (1), ethyl gallate (9) and (+)-dehydrovomifoliol (12) exhibited inhibitory activity on the nitric oxide production of RAW 264.7 at a concentration of 25 μM. The result indicated that the combination HSCCC with pre-HPLC is an effective way for compound separation and purification. And Diaphragma juglandis fructus constituents have the potential for the treatment of inflammatory-related diseases

Deep learning in image-based breast and cervical cancer detection:a systematic review and meta-analysis

Accurate early detection of breast and cervical cancer is vital for treatment success. Here, we conduct a meta-analysis to assess the diagnostic performance of deep learning (DL) algorithms for early breast and cervical cancer identification. Four subgroups are also investigated: cancer type (breast or cervical), validation type (internal or external), imaging modalities (mammography, ultrasound, cytology, or colposcopy), and DL algorithms versus clinicians. Thirty-five studies are deemed eligible for systematic review, 20 of which are meta-analyzed, with a pooled sensitivity of 88% (95% CI 85–90%), specificity of 84% (79–87%), and AUC of 0.92 (0.90–0.94). Acceptable diagnostic performance with analogous DL algorithms was highlighted across all subgroups. Therefore, DL algorithms could be useful for detecting breast and cervical cancer using medical imaging, having equivalent performance to human clinicians. However, this tentative assertion is based on studies with relatively poor designs and reporting, which likely caused bias and overestimated algorithm performance. Evidence-based, standardized guidelines around study methods and reporting are required to improve the quality of DL research

PREPARATIVE SEPARATION OF ISOQUINOLINE ALKALOIDS FROM PHELLODENDRI CORTEX BY PH-ZONE-REFINING COUNTER-CURRENT CHROMATOGRAPHY

pH-Zone-refining counter-current chromatography (PZRCCC) was successfully used for preparative separation isoquinoline alkaloids from Phellodendri Cortex. Two target compounds of berberine and palmatine were completely resolved by a two-phase solvent system chloroform/methanol/water (4:2:2, v/v), with 10 mmol L-1 HCl in upper stationary phase and 10 mmol L-1 TEA in lower organic phase. From 2.0 g of the crude alkaloids, palmatine (448 mg, 95.1%) and berberine (159 mg, 95.4%) were obtained in one-step PZRCCC separation. The chemical structures were identified by ESI-MS and NMR data. The results indicated that PZRCCC separation is an excellent preparative separation method for isoquinoline alkaloids from Phellodendri Cortex

PREPARATIVE SEPARATION OF ISOQUINOLINE ALKALOIDS FROM PHELLODENDRI CORTEX BY PH-ZONE-REFINING COUNTER-CURRENT CHROMATOGRAPHY

<div><p>pH-Zone-refining counter-current chromatography (PZRCCC) was successfully used for preparative separation isoquinoline alkaloids from Phellodendri Cortex. Two target compounds of berberine and palmatine were completely resolved by a two-phase solvent system chloroform/methanol/water (4:2:2, v/v), with 10 mmol L-1 HCl in upper stationary phase and 10 mmol L-1 TEA in lower organic phase. From 2.0 g of the crude alkaloids, palmatine (448 mg, 95.1%) and berberine (159 mg, 95.4%) were obtained in one-step PZRCCC separation. The chemical structures were identified by ESI-MS and NMR data. The results indicated that PZRCCC separation is an excellent preparative separation method for isoquinoline alkaloids from Phellodendri Cortex.</p></div