Estrogen Acts Through Estrogen Receptor 2b to Regulate Hepatobiliary Fate During Vertebrate Development.

BACKGROUND AND AIMS: During liver development, bipotent progenitor cells differentiate into hepatocytes and biliary epithelial cells to ensure a functional liver required to maintain organismal homeostasis. The developmental cues controlling the differentiation of committed progenitors into these cell types, however, are incompletely understood. Here, we discover an essential role for estrogenic regulation in vertebrate liver development to affect hepatobiliary fate decisions. APPROACH AND RESULTS: Exposure of zebrafish embryos to 17ß-estradiol (E2) during liver development significantly decreased hepatocyte-specific gene expression, liver size, and hepatocyte number. In contrast, pharmacological blockade of estrogen synthesis or nuclear estrogen receptor (ESR) signaling enhanced liver size and hepatocyte marker expression. Transgenic reporter fish demonstrated nuclear ESR activity in the developing liver. Chemical inhibition and morpholino knockdown of nuclear estrogen receptor 2b (esr2b) increased hepatocyte gene expression and blocked the effects of E2 exposure. esr2b-/- mutant zebrafish exhibited significantly increased expression of hepatocyte markers with no impact on liver progenitors, other endodermal lineages, or vasculature. Significantly, E2-stimulated Esr2b activity promoted biliary epithelial differentiation at the expense of hepatocyte fate, whereas loss of esr2b impaired biliary lineage commitment. Chemical and genetic epistasis studies identified bone morphogenetic protein (BMP) signaling as a mediator of the estrogen effects. The divergent impact of estrogen on hepatobiliary fate was confirmed in a human hepatoblast cell line, indicating the relevance of this pathway for human liver development. CONCLUSIONS: Our studies identify E2, esr2b, and downstream BMP activity as important regulators of hepatobiliary fate decisions during vertebrate liver development. These results have significant clinical implications for liver development in infants exposed to abnormal estrogen levels or estrogenic compounds during pregnancy.

Antihypertensive Effects of Corn Silk Extract and Its Novel Bioactive Constituent in Spontaneously Hypertensive Rats: The Involvement of Angiotensin-Converting Enzyme Inhibition.

Corn silk tea has been used in folk medicine for anti-hypertensive healthcare. Angiotensin-converting enzyme (ACE) plays a crucial role on the homeostasis of blood pressure. However, effects of corn silk tea on ACE activity and the presence of ACE inhibitory constituents in corn silk are still unknown. Here we applied proteomics and bioinformatics approaches to identify corn silk bioactive peptides (CSBps) that target ACE from the boiling water extract of corn silk (CSE). CSE significantly reduced systolic blood pressure (SBP) levels in spontaneously hypertensive rats and inhibited the ACE activity. By proteomics coupled with bioinformatics analyses, we identified a novel ACE inhibitory peptide CSBp5 in CSE. CSBp5 significantly inhibited the ACE activity and decreased SBP levels in a dose-dependent manner. Docking analysis showed that CSBp5 occupied the substrate-binding channel of ACE and interacted with ACE via hydrogen bonds. In conclusion, we identified that CSE exhibited anti-hypertensive effects in SHRs via the inhibition of ACE, the target of most anti-hypertensive drugs. In addition, an ACE inhibitory phytopeptide CSBp5 that decreased SBP levels in rats was newly identified. Our findings supported the ethnomedical use of corn silk tea on hypertension. Moreover, the identification of ACE inhibitory phytopeptide in corn silk further strengthened our findings.

Primordial odontogenic tumour: report of two cases.

AIMS: Primordial odontogenic tumour (POT) is a rare mixed odontogenic neoplasm that is composed of primitive ectomesenchyme resembling dental papilla, surfaced by odontogenic epithelium resembling inner enamel epithelium, without hard tissue formation. Most reported cases have presented in the posterior mandible as a well-demarcated radiolucency associated with an unerupted tooth in the first two decades of life. The aim of this report is to describe the clinicopathological features of two more cases of POT. METHODS AND RESULTS: Each presented as an asymptomatic well-delineated radiolucency in the mandible in a 15-year-old female and an 18-year-old male, respectively. Both tumours were composed of a proliferation of plump spindle and stellate cells in delicately collagenous and myxoid stroma, surfaced by columnar-squamous epithelial cells with reverse nuclear polarisation at the tumour periphery. In one case, the formation of abortive tooth germ-like structures was noted. This has not been reported previously and supports the hypothesis of the primordial nature of this tumour. Both patients showed no recurrence at 3- and 20-month follow-up, respectively. CONCLUSION: This report describes two additional cases of POT for a total of 11 cases reported in the English language literature.

Comprehensive evaluation of gene expression signatures in response to electroacupuncture stimulation at Zusanli (ST36) acupoint by transcriptomic analysis.

BACKGROUND: Electroacupuncture (EA) has been applied to treat and prevent diseases for years. However, molecular events happened in both the acupunctured site and the internal organs after EA stimulation have not been clarified. METHODS: Here we applied transcriptomic analysis to explore the gene expression signatures after EA stimulation. Mice were applied EA stimulation at ST36 for 15 min and nine tissues were collected three hours later for microarray analysis. RESULTS: We found that EA affected the expression of genes not only in the acupunctured site but also in the internal organs. EA commonly affected biological networks involved in cytoskeleton and cell adhesion, and also regulated unique process networks in specific organs, such as γ-aminobutyric acid-ergic neurotransmission in brain and inflammation process in lung. In addition, EA affected the expression of genes related to various diseases, such as neurodegenerative diseases in brain and obstructive pulmonary diseases in lung. CONCLUSIONS: This report applied, for the first time, a global comprehensive genome-wide approach to analyze the gene expression profiling of acupunctured site and internal organs after EA stimulation. The connection between gene expression signatures, biological processes, and diseases might provide a basis for prediction and explanation on the therapeutic potentials of acupuncture in organs.

Hypoglycemic effects of Trichosanthes kirilowii and its protein constituent in diabetic mice: the involvement of insulin receptor pathway.

BACKGROUND: Diabetes is a serious chronic metabolic disorder. Trichosanthes kirilowii Maxim. (TK) is traditionally used for the treatment of diabetes in traditional Chinese medicine (TCM). However, the clinical application of TK on diabetic patients and the hypoglycemic efficacies of TK are still unclear. METHODS: A retrospective cohort study was conducted to analyze the usage of Chinese herbs in patients with type 2 diabetes in Taiwan. Glucose tolerance test was performed to analyze the hypoglycemic effect of TK. Proteomic approach was performed to identify the protein constituents of TK. Insulin receptor (IR) kinase activity assay and glucose tolerance tests in diabetic mice were further used to elucidate the hypoglycemic mechanisms and efficacies of TK. RESULTS: By a retrospective cohort study, we found that TK was the most frequently used Chinese medicinal herb in type 2 diabetic patients in Taiwan. Oral administration of aqueous extract of TK displayed hypoglycemic effects in a dose-dependent manner in mice. An abundant novel TK protein (TKP) was further identified by proteomic approach. TKP interacted with IR by docking analysis and activated the kinase activity of IR. In addition, TKP enhanced the clearance of glucose in diabetic mice in a dose-dependent manner. CONCLUSIONS: In conclusion, this study applied a bed-to-bench approach to elucidate the hypoglycemic efficacies and mechanisms of TK on clinical usage. In addition, we newly identified a hypoglycemic protein TKP from TK. Our findings might provide a reasonable explanation of TK on the treatment of diabetes in TCM.

Microfabrication of Cell-Laden Hydrogels for Engineering Mineralized and Load Bearing Tissues.

Microengineering technologies and advanced biomaterials have extensive applications in the field of regenerative medicine. In this chapter, we review the integration of microfabrication techniques and hydrogel-based biomaterials in the field of dental, bone, and cartilage tissue engineering. We primarily discuss the major features that make hydrogels attractive candidates to mimic extracellular matrix (ECM), and we consider the benefits of three-dimensional (3D) culture systems for tissue engineering applications. We then focus on the fundamental principles of microfabrication techniques including photolithography, soft lithography and bioprinting approaches. Lastly, we summarize recent research on microengineering cell-laden hydrogel constructs for dental, bone and cartilage regeneration, and discuss future applications of microfabrication techniques for load-bearing tissue engineering.

Rare mandibular surgical ciliated cysts: report of two new cases.

The surgical ciliated cyst, also known as the "postoperative maxillary cyst" or "implantation cyst," occurs as a result of iatrogenic implantation of respiratory epithelium into a noncontiguous surgical site after sinus surgery. It typically presents as a well-defined radiolucency in the maxilla in young adults. Histopathologically, the cyst is lined by ciliated columnar, cuboidal, or pseudostratified squamous epithelium with mucous cells. We report two rare cases of a surgical ciliated cyst located in the mandible.

Evodiamine inhibits 12-O-tetradecanoylphorbol-13-acetate-induced activator protein 1 transactivation and cell transformation in human hepatocytes.

Evodia rutaecarpa has been used to treat inflammatory digestive disorders in Asian countries. However, little is known about the antitumor activities of E. rutaecarpa and its bioactive constituent evodiamine (EVO). The aim of this study was to characterize the antitumor mechanisms of E. rutaecarpa and EVO in human hepatocytes. Human Chang liver cells were transfected with activator protein 1 (AP-1)-luciferase reporter gene and designated as Chang/AP-1 cells. The Chang/AP-1 cells were treated with E. rutaecarpa and its bioactive constituents, and challenged with the AP-1 stimulator 12-O-tetradecanoylphorbol-13- acetate (TPA). The present study showed that the methanol extract of E. rutaecarpa decreased the TPA-induced AP-1 transactivation in Chang/AP-1 cells, with an EC50 value of 24.72 µg/mL. EVO inhibited the TPA-induced AP-1 transactivation and colony formation, with EC50 values of 82 µM and 8.2 µM, respectively. Moreover, EVO significantly diminished the TPA-induced phosphorylation of extracellular signal-regulated kinases (ERKs). These results suggested that EVO treatment suppressed the TPA-induced AP-1 activity via the ERKs pathway. In conclusion, EVO inhibited the AP-1 activity and cellular transformation in human hepatocytes, suggesting that EVO was a potential agent for antitumor therapy.

Architectural Alterations in Oral Epithelial Dysplasia are Similar in Unifocal and Proliferative Leukoplakia.

The current WHO histopathologic criteria for oral epithelial dysplasia (ED) are based on architectural and cytologic alterations, and do not address other histopathologic features of ED. Here we propose new diagnostic criteria including architectural, organizational, and cytologic features for oral ED. Cases of unifocal leukoplakia (UL) and proliferative leukoplakia (PL) with clinical photographs and follow-up information were identified. Only cases that showed minimal cytologic atypia or mild ED were used to demonstrate critical architectural changes as defined in this study. Eight biopsies from eight UL patients and 34 biopsies from four PL patients were included. The biopsies showed (a) corrugated, verrucous or papillary architecture, (b) hyperkeratosis with epithelial atrophy, (c) bulky squamous epithelial proliferation, and (d) demarcated hyperkeratosis and "skip" segments. The architectural alterations defined here are as important as the currently used criteria for the diagnosis of ED. Clinicopathologic correlation when diagnosing oral ED is also of the utmost importance in accurate diagnosis.

Conserved charged amino acid residues in the extracellular region of sodium/iodide symporter are critical for iodide transport activity.

BACKGROUND: Sodium/iodide symporter (NIS) mediates the active transport and accumulation of iodide from the blood into the thyroid gland. His-226 located in the extracellular region of NIS has been demonstrated to be critical for iodide transport in our previous study. The conserved charged amino acid residues in the extracellular region of NIS were therefore characterized in this study. METHODS: Fourteen charged residues (Arg-9, Glu-79, Arg-82, Lys-86, Asp-163, His-226, Arg-228, Asp-233, Asp-237, Arg-239, Arg-241, Asp-311, Asp-322, and Asp-331) were replaced by alanine. Iodide uptake abilities of mutants were evaluated by steady-state and kinetic analysis. The three-dimensional comparative protein structure of NIS was further modeled using sodium/glucose transporter as the reference protein. RESULTS: All the NIS mutants were expressed normally in the cells and targeted correctly to the plasma membrane. However, these mutants, except R9A, displayed severe defects on the iodide uptake. Further kinetic analysis revealed that mutations at conserved positively charged amino acid residues in the extracellular region of NIS led to decrease NIS-mediated iodide uptake activity by reducing the maximal rate of iodide transport, while mutations at conserved negatively charged residues led to decrease iodide transport by increasing dissociation between NIS mutants and iodide. CONCLUSIONS: This is the first report characterizing thoroughly the functional significance of conserved charged amino acid residues in the extracellular region of NIS. Our data suggested that conserved charged amino acid residues, except Arg-9, in the extracellular region of NIS were critical for iodide transport.

Benign Alveolar Ridge Keratosis: Clinical and Histopathologic Analysis of 167 Cases.

Benign alveolar ridge keratosis (BARK), the intraoral counterpart of cutaneous lichen simplex chronicus, is a reactive hyperkeratosis caused by trauma or friction that presents as a poorly demarcated white papule or plaque on the keratinized mucosa of the retromolar pad or alveolar ridge mucosa (often edentulous). This is a clinical and histopathologic analysis of BARK including evaluation of p53 expression in selected cases. One hundred and sixty-seven cases of BARK were identified from 2016 to 2017 and 112 (67.1%) occurred in males with a median age of 56 years (range 15-86). The retromolar pad was affected in 107 (64.1%) cases and the edentulous alveolar mucosa in 60 (35.9%) cases, with 17.4% of the cases presenting bilaterally. BARK showed hyperkeratosis often with wedge-shaped hypergranulosis and occasional focal parakeratosis. The epithelium exhibited acanthosis and surface corrugation with tapered rete ridges often interconnected at the tips. The study for p53 performed in 12 cases showed less than 25% nuclear positivity. BARK is a distinct benign clinicopathologic entity caused by friction, which should be clearly distinguished from true leukoplakia, a potentially malignant disorder.

Oral Cancer: Genetics and the Role of Precision Medicine.

Oral squamous cell carcinoma (OSCC) is one of the leading cancers in the world. OSCC patients are managed with surgery and/or chemoradiation. Prognoses and survival rates are dismal, however, and have not improved for more than 20 years. Recently, the concept of precision medicine was introduced, and the introduction of targeted therapeutics demonstrated promising outcomes. This article reviews the current understanding of initiation, progression, and metastasis of OSCC from both genetic and epigenetic perspectives. In addition, the applications and integration of omics technologies in biomarker discovery and drug development for treating OSCC are reviewed.

Vanillin improves and prevents trinitrobenzene sulfonic acid-induced colitis in mice.

Inflammatory bowel disease (IBD) is chronic inflammatory and relapsing disease of the gut. It has been known that activation of nuclear factor-kappaB (NF-kappaB) and production of proinflammatory cytokines play important roles in the pathogenesis of IBD. In this study, the effect of vanillin (4-hydroxy-3-methoxybenzaldehyde), a potent nuclear factor-kappaB (NF-kappaB) inhibitor, was evaluated in mice with trinitrobenzene sulfonic acid (TNBS)-induced colitis. Oral administration of vanillin improved macroscopic and histological features of TNBS-induced colitis in a dose-dependent manner. Vanillin not only prevented TNBS-induced colitis but also ameliorated the established colitis. By in vivo NF-kappaB bioluminescence imaging, electrophoretic mobility shift assay, and Western blot, we found that vanillin suppressed in vivo NF-kappaB activities through the inhibition of p65 translocation, inhibitor of nuclear factor-kappaB(IkappaB)-alpha phosphorylation, and IkappaB kinase activation. Furthermore, vanillin reduced the expressions of proinflammatory cytokines [interleukin (IL)-1beta, IL-6, interferon-gamma, and tumor necrosis factor-alpha] and stimulated the expression of anti-inflammatory cytokine (IL-4) in colonic tissues. In conclusion, this work identified vanillin as an anti-inflammatory compound with the capacity to prevent and ameliorate TNBS-induced colitis. Due to its safety, vanillin could be a potent candidate for the treatment of IBD.

Mutagenesis identifies the critical regions and amino acid residues of suid herpesvirus 1 DNA-binding protein required for DNA binding and strand invasion.

Herpesviral DNA-binding protein (DBP) is a unique protein involved in viral DNA replication and genomic recombination. It binds and stabilizes the single-stranded DNA. It also forms the D-loops and promotes the strand invasion. To identify the functional regions and amino acid residues required for DNA binding and D-loop formation, we characterized several DBP mutants of suid herpesvirus 1 (SuHV-1). Acetic anhydride modification assay showed that lysine residues were critical for DNA binding and D-loop formation. Replacement of highly conserved lysine residues with alanine revealed that Lys-756 and Lys-970 were critical for DNA binding, while Lys-161 participated in DNA binding and D-loop formation. Analysis of nested deleted mutants showed that N-terminal 201 amino acid residues and C-terminal 305 amino acid residues were required for D-loop formation and DNA binding, respectively. In conclusion, these findings suggested that SuHV-1 DBP contained critical regions for DNA binding and D-loop formation, and Lys-161, Lys-756, and Lys-970 were required for DNA binding or D-loop formation.

Mutagenesis identifies the critical amino acid residues of human endonuclease G involved in catalysis, magnesium coordination, and substrate specificity.

BACKGROUND: Endonuclease G (EndoG), a member of DNA/RNA nonspecific betabetaalpha-Me-finger nucleases, is involved in apoptosis and normal cellular proliferation. In this study, we analyzed the critical amino acid residues of EndoG and proposed the catalytic mechanism of EndoG. METHODS: To identify the critical amino acid residues of human EndoG, we replaced the conserved histidine, asparagine, and arginine residues with alanine. The catalytic efficacies of Escherichia coli-expressed EndoG variants were further analyzed by kinetic studies. RESULTS: Diethyl pyrocarbonate modification assay revealed that histidine residues were involved in EndoG activity. His-141, Asn-163, and Asn-172 in the H-N-H motif of EndoG were critical for catalysis and substrate specificity. H141A mutant required a higher magnesium concentration to achieve its activity, suggesting the unique role of His-141 in both catalysis and magnesium coordination. Furthermore, an additional catalytic residue (Asn-251) and an additional metal ion binding site (Glu-271) of human EndoG were identified. CONCLUSION: Based on the mutational analysis and homology modeling, we proposed that human EndoG shared a similar catalytic mechanism with nuclease A from Anabaena.

Crohn's disease initiated with extraintestinal features.

Crohn's disease (CD) is a multifactorial, chronic immune-mediated disorder. The oral cavity is involved in 0.5% to 20% of the patients with CD. Oral manifestations of CD are sometimes nonspecific and can be overlooked by the clinicians. These manifestations may precede intestinal symptoms and can serve as indicators for early diagnosis. To increase awareness and to contribute to the standard intervention, here we report a paediatric case with persistent idiopathic swelling of the lower lip and perianal fistula. Microscopic examinations revealed multiple non-necrotising granulomas with chronic inflammation, oedema and lymphangiectasia. The patient was treated with metronidazole 500 mg and ciprofloxacin 500 mg twice a day for one month. The perioral lesions were managed with topical 0.03% tacrolimus and oral prednisone 10 mgtwice a day for one month, followed by a tapered regimen of 10 mg/day for another month. The patient's symptoms improved without full remission at the 6-month follow-up.

Ferulic acid altered IL-17A/IL-17RA interaction and protected against imiquimod-induced psoriasis-like skin injury in mice.

Ferulic acid (FA), a phenolic phytochemical, is commonly found in grains, vegetables, and fruits. Interleukin-17A (IL-17A) and IL-17 receptor A (IL-17RA) interaction is one of important therapeutic targets for psoriasis. Here we analyzed the FA effects on IL-17A/IL-17RA interaction and psoriasis-like skin injury induced by imiquimod (IMQ). IL-17A-blocking assay and docking analysis showed that FA interacted with Trp-67, Gln-94, and Glu-95 residues of IL-17A via hydrogen bonds and consequently abolished the binding of IL-17RA to IL-17A. Mice were topically given with IMQ and orally given with various amounts of FA for 14 consecutive days. FA attenuated IMQ-induced psoriasis-like skin lesions in a dose-dependent manner, and the epidermal thickness of mice treated with 100 mg/kg FA was reduced by 53.48 ±â€¯4.44% in comparison with sham. Global analysis of differentially expressed genes showed that IMQ and FA significantly affected immune response, metabolism, and mitogen-activated protein kinase signaling pathways. Immunohistochemical staining showed that FA inhibited the infiltration and the cytokine secretion of Th17 cell, dendritic cell, and granulocyte subsets in psoriatic skin tissues. In conclusion, we newly identified that oral administration of FA protected against IMQ-induced psoriatic skin injury in mice. Moreover, its protection was associated with the interference of IL-17A/IL-17RA interaction.

Eriobotrya japonica leaf and its triterpenes inhibited lipopolysaccharide-induced cytokines and inducible enzyme production via the nuclear factor-kappaB signaling pathway in lung epithelial cells.

Pulmonary inflammation is a characteristic of many lung diseases. Increased levels of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and proinflammatory cytokines, such as interleukin-1beta (IL-1beta), tumor necrosis factor-alpha (TNF-alpha) and IL-8, have been correlated with lung inflammation. In this study, we used lipopolysaccharide (LPS) to induce iNOS, COX-2, and cytokines (TNF-alpha, IL-1beta, and IL-8) productions in human lung epithelial cells (A-549). Leaf of Eriobotrya japonica (Pi-Pa-Ye, PPY), a traditional Chinese medicine for the treatment of pulmonary inflammatory diseases, was capable of suppressing LPS-induced cytokine productions in a dose-dependent manner. Moreover, the suppression of PPY on the cytokine productions resulted from the inhibition of inhibitory kappaB-alpha phosphorylation and nuclear factor-kappaB (NF-kappaB) activation. Analysis of the anti-inflammatory effects of ursolic acid and oleanolic acid, the triterpene compounds present in PPY, showed that ursolic acid significantly inhibited LPS-induced IL-8 production, NF-kappaB activation, and iNOS mRNA expression, whereas oleanolic acid did not have these effects. In conclusion, our findings suggested the potential mechanisms of PPY and its active component, ursolic acid, in the treatment of pulmonary inflammation.

Relationship Between San-Huang-Xie-Xin-Tang and its herbal components on the gene expression profiles in HepG2 cells.

Traditional Chinese medicine (TCM) has been used for thousands of years. Most Chinese herbal formulae consist of several herbal components and have been used to treat various diseases. However, the mechanisms of most formulae and the relationship between formulae and their components remain to be elucidated. Here we analyzed the putative mechanism of San-Huang-Xie-Xin-Tang (SHXXT) and defined the relationship between SHXXT and its herbal components by microarray technique. HepG2 cells were treated with SHXXT or its components and the gene expression profiles were analyzed by DNA microarray. Gene set enrichment analysis indicated that SHXXT and its components displayed a unique anti-proliferation pattern via p53 signaling, p53 activated, and DNA damage signaling pathways in HepG2 cells. Network analysis showed that most genes were regulated by one molecule, p53. In addition, hierarchical clustering analysis showed that Rhizoma Coptis shared a similar gene expression profile with SHXXT. These findings may explain why Rhizoma Coptis is the principle herb that exerts the major effect in the herbal formula, SHXXT. Moreover, this is the first report to reveal the relationship between formulae and their herbal components in TCM by microarray and bioinformatics tools.

Oral Cancer: Genetics and the Role of Precision Medicine.

Oral squamous cell carcinoma (OSCC) is one of the leading cancers in the world. OSCC patients are managed with surgery and/or chemoradiation. Prognoses and survival rates are dismal, however, and have not improved for more than 20 years. Recently, the concept of precision medicine was introduced, and the introduction of targeted therapeutics demonstrated promising outcomes. This article reviews the current understanding of initiation, progression, and metastasis of OSCC from both genetic and epigenetic perspectives. In addition, the applications and integration of omics technologies in biomarker discovery and drug development for treating OSCC are reviewed.