Search

Background: Studies have reported clinical heterogeneity between right-sided colon cancer (RCC) and left-sided colon cancer (LCC). However, none of these studies used multi-omics analysis combining genetic regulation, microbiota, and metabolites to explain the site-specific difference., Methods: Here, 494 participants from a 16S rRNA gene sequencing cohort (50 RCC, 114 LCC, and 100 healthy controls) and a multi-omics cohort (63 RCC, 79 LCC, and 88 healthy controls) were analyzed. 16S rRNA gene, metagenomic sequencing, and metabolomics analyses of fecal samples were evaluated to identify tumor location-related bacteria and metabolites. Whole-exome sequencing (WES) and transcriptome sequencing (RNA-seq) were conducted to obtain the mutation burden and genomic expression pattern., Results: We found unique profiles of the intestinal microbiome, metabolome, and host genome between RCC and LCC. The bacteria Flavonifractor plautii (Fp) and Fusobacterium nucleatum, the metabolites L-phenylalanine, and the host genes PHLDA1 and WBP1 were the key omics features of RCC; whereas the bacteria Bacteroides sp. A1C1 (B.A1C1) and Parvimonas micra, the metabolites L-citrulline and D-ornithine, and the host genes TCF25 and HLA-DRB5 were considered the dominant omics features in LCC. Multi-omics correlation analysis indicated that RCC-enriched Fp was related to the accumulation of the metabolite L-phenylalanine and the suppressed WBP1 signal in RCC patients. In addition, LCC-enriched B.A1C1 was associated with the accumulation of the metabolites D-ornithine and L-citrulline as well as activation of the genes TCF25, HLA-DRB5, and AC079354.1., Conclusion: Our findings identify previously unknown links between intestinal microbiota alterations, metabolites, and host genomics in RCC vs. LCC, suggesting that it may be possible to treat colorectal cancer (CRC) by targeting the gut microbiota-host interaction. Video Abstract., Competing Interests: Declarations. Ethics approval and consent to participate: Ethical approval was obtained from the Institutional Review Board of Fudan University Shanghai Cancer Center (ID: 050432–4-1911D), and written informed consent was provided by all subjects before sampling. Consent for publication: Not applicable. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

Periodontitis seriously affects oral health worldwide. Despite extensive efforts in prevention and treatment methods over the years, the prevalence of periodontitis in the population has not decreased. DNA damage-induced cellular senescence may be one of the mechanisms underlying periodontitis.Sirtuin7 (SIRT7) has deacetylase activity and regulates a variety of biological processes, including cell proliferation, death, and DNA damage repair.Increasing evidence confirms the crucial role of SIRT7 in age-related and inflammatory diseases. However, the mechanism of action of SIRT7 in periodontitis remains unclear. Our study demonstrates that SIRT7 is downregulated in human periodontal ligament fibroblasts induced by Porphyromonas gingivalis lipopolysaccharide (Pg-LPS). Overexpression of the SIRT7 gene significantly reduces the production of senescence-related molecules P53, P21, P16, as well as inflammatory cytokines IL-1β and TNF-α stimulated by Pg-LPS. Furthermore, overexpression of the SIRT7 gene significantly decreases the phosphorylation levels of AKT and mTOR in Pg-LPS-treated hPDLFs. Conversely, SIRT7 gene knockdown exhibits opposite effects compared to overexpression in Pg-LPS-treated hPDLFs. In conclusion, our findings indicate that SIRT7 can inhibit Pg-LPS-induced senescence and consequently suppress the secretion of inflammatory cytokines through the AKT/mTOR pathway. As a result, SIRT7 could be regarded a viable pharmaceutical target for clinical periodontitis treatment., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Intestinal stem cells are crucial for maintaining intestinal homeostasis, yet their transformation into tumor stem cells in the context of microbial infection remains poorly understood. Fusobacterium nucleatum (F. nucleatum) is frequently associated with the onset and progression of colorectal cancer (CRC). In this study, we uncovered that F. nucleatum colonized the depths of gut crypts in both human CRC patients and mouse models. Through single-cell sequencing analysis, we demonstrated that F. nucleatum infection reprogrammed crypt cells and activated LY6A+ revival stem cells (RSCs), promoting their hyperproliferation and subsequent transformation into tumor stem cells, which accelerated intestinal carcinogenesis. Mechanistically, we identified LY6A as a GPI-anchored membrane receptor for F. nucleatum. Upon binding, F. nucleatum induced upregulation of RPS14 via the LY6A receptor, driving RSC hyperactivity and tumorigenic conversion. Functional studies showed that genetic ablation of Ly6a in intestinal epithelial cells or Rps14 in LY6A+ RSCs substantially reduced F. nucleatum colonization and tumorigenesis. Moreover, clinical CRC cohorts analysis revealed a strong correlation between F. nucleatum infection, RSC expansion, and elevated RPS14 expression in tumor tissues. These findings highlight an alternative F. nucleatum-LY6A-RPS14 signaling axis as a critical driver of CRC progression and propose potential therapeutic targets for effective CRC intervention.

An imbalanced system of angiogenesis-osteoblasts-osteoclasts is regarded as the main factor in bone remodeling dysfunction diseases or osseointegration loss. Osteoclast precursors are the key cells that accelerate bone-specific angiogenesis and maintain normal osteoblast and osteoclast function. Graphene oxide is an effective scaffold surface modification agent with broad application prospects in bone tissue engineering. However, the effect of graphene oxide on the interaction between osteoclasts and angiogenesis has not yet been elucidated. In this study, a rat calvarial defect model was established and treated with an electrochemically derived nanographene oxide (ENGO) hydrogel. Higher angiogenesis and platelet-derived growth factor (PDGF) B in preosteoclasts were observed in the ENGO group compared with that in the control group. Moreover, in vitro experiments demonstrate the efficacy of ENGO in substantially reducing the expression of the receptor activator of nuclear factor-kappaB ligand (RANKL)-induced osteoclast-associated markers and inhibiting bone resorption activity. Additionally, ENGO enhances the secretion of the osteoclast-derived coupling factor PDGF-BB and promotes angiogenesis. Our investigation revealed the crucial role of isocitrate dehydrogenase 1 (IDH1) in the ENGO-mediated regulation of osteoclast differentiation and PDGF-BB secretion. The decreased expression of IDH1 reduces the level of histone lysine demethylase 7A (KDM7A) and subsequently increases the H3K9me2 level in the cathepsin K promoter region. In summary, we found that ENGO promotes angiogenesis by inhibiting the maturity of RANKL-induced osteoclasts and enhancing PDGF-BB secretion. These results indicate that ENGO holds promise for the application in fostering osteoclast-endothelial cell crosstalk, providing an effective strategy for treating bone resorption and osteoclast-related bone loss diseases.

The etiology of gastrointestinal (GI) diseases is intricate and multifactorial, encompassing complex interactions between genetic predisposition and gut microbiota. The cell fate change, immune function regulation, and microenvironment composition in diseased tissues are governed by microorganisms and mutated genes either independently or through synergistic interactions. A comprehensive understanding of GI disease etiology is imperative for developing precise prevention and treatment strategies. However, the existing models used for studying the microenvironment in GI diseases-whether cancer cell lines or mouse models-exhibit significant limitations, which leads to the prosperity of organoids models. This review first describes the development history of organoids models, followed by a detailed demonstration of organoids application from bench to clinic. As for bench utilization, we present a layer-by-layer elucidation of organoid simulation on host-microbial interactions, as well as the application in molecular mechanism analysis. As for clinical adhibition, we provide a generalized interpretation of organoid application in GI disease simulation from inflammatory disorders to malignancy diseases, as well as in GI disease treatment including drug screening, immunotherapy, and microbial-targeting and screening treatment. This review draws a comprehensive and systematical depiction of organoids models, providing a novel insight into the utilization of organoids models from bench to clinic., Competing Interests: The authors declare no conflict of interest., (© 2024 The Author(s). MedComm published by Sichuan International Medical Exchange & Promotion Association (SCIMEA) and John Wiley & Sons Australia, Ltd.)

Obesity is a risk factor for colorectal cancer (CRC), and the involvement of gut microbiota in the pathogenesis of obesity and CRC is widely recognized. However, the landscape of fecal microbiome and metabolome distinguishing patients with obesity-related CRC from obesity remains unknown. Here, we utilize metagenomic sequencing and metabolomics from 522 patients with CRC and healthy controls to identify the characteristics of obese CRC. Our integrated analysis reveals that obesity-related CRC is characterized by elevated Peptostreptococcus stomatis, dysregulated fatty acids and phospholipids, and altered Kyoto Encyclopedia of Genes and Genomes pathways involving glycerophospholipid metabolism and lipopolysaccharide synthesis. Correlation analysis unveils microbial interactions in obesity, where the probiotic Faecalibacterium prausnitzii and the tumor-promoting species P. stomatis may engage in cross-feeding, thereby promoting tumorigenesis. In vitro experiments affirm enhanced growth under cross-feeding conditions. The mutualistic microbe-microbe interaction may contribute to the association between obesity and elevated CRC risk. Additionally, diagnostic models incorporating BMI-specific microbial biomarkers display promise for precise CRC screening., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

Japanese encephalitis (JE) is a naturally occurring localized disease caused by the Japanese encephalitis virus, which is spread by the Culex tritaeniorhynchus. China has a high rate of JE. Shanxi, located in North China, has a high prevalence of adult JE. Adult JE has more severe complications, mortality, and a higher disease burden, making it a public health issue. This retrospective study examined the dynamic epidemic changes, high-risk areas of JE, and clinical characteristics and prognostic factors of adult JE in Shanxi Province. The findings revealed that July to September was the primary epidemic season of JE and that JE cases were mainly in individuals over the age of 40. The incidence of JE from 2005 to 2022 demonstrated a positive spatial correlation with significant clustering characteristics, with high-incidence clusters in the south and southeast. Multivariate logistic regression analysis revealed that higher cerebrospinal fluid pressure, higher white blood cell counts, higher neutrophil percentage, deep coma, and lower albumin were independent factors for poor prognosis of adult JE. The developed risk prediction model holds great promise in early prognosis assessment of patients, providing a basis for clinical decision-making and early clinical intervention., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Zheng, Wen, Liu and Wang.)

Background: Endoplasmic reticulum stress (ER stress) is activated in chronic sinusitis with nasal polyps (CRSwNP) and leads to increased expression of C/EBP homologous protein (CHOP). However, the role of CHOP in the pathogenesis of CRSwNP remains unclear., Methods: CHOP expression was detected by immunohistochemistry staining in nasal mucosa of control and CRSwNP patients. Co-localization of CHOP and cleaved caspase3, p-MLKL, and CD68 was detected by immunofluorescence staining in CRSwNP patients. TNFα, IFNγ, IL1β, LPS, and tunicamycin were added to primary dispersed nasal polyp cells (DNPCs) to explore their roles in cell death. Western blot, CCK8 assay, and flow cytometry were employed to detect cell death., Results: CHOP was specifically activated in CRSwNP compared to controls. It was mainly macrophages that highly expressed CHOP, some of which underwent apoptosis and the other underwent necroptosis. IL1β induced increased CHOP and apoptosis, and a slight p-MLKL. In addition, ER stress could also promote p-MLKL expression. Whereas classical TNFα plus IFNγ and LPS did not induce increased necroptosis in DNPCs., Conclusion: IL1β induced the apoptotic pathway and minor necroptosis. And ER stress also plays a role in the occurrence of necroptosis in CRSwNP.

Chronic hepatitis B virus (HBV) infection is a major global public health risk that threatens human life and health, although the number of vaccinated people has increased. The clinical outcome of HBV infection depends on the complex interplay between viral replication and the host immune response. Innate immunity plays an important role in the early stages of the disease but retains no long-term immune memory. However, HBV evades detection by the host innate immune system through stealth. Therefore, adaptive immunity involving T and B cells is crucial for controlling and clearing HBV infections that lead to liver inflammation and damage. The persistence of HBV leads to immune tolerance owing to immune cell dysfunction, T cell exhaustion, and an increase in suppressor cells and cytokines. Although significant progress has been made in HBV treatment in recent years, the balance between immune tolerance, immune activation, inflammation, and fibrosis in chronic hepatitis B remains unknown, making a functional cure difficult to achieve. Therefore, this review focuses on the important cells involved in the innate and adaptive immunity of chronic hepatitis B that target the host immune system and identifies treatment strategies., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Zheng, Dou and Wang.)

Objective: To explore the potential osteogenic induction mechanism of enamel matrix derivatives (EMDs) on bone marrow mesenchymal stem cell (BMSC) sheets with different titanium surface morphologies., Methods: The BMSCs were inoculated on the surfaces of titanium alloys with different morphologies: anodic oxidation (AO), sand-blasted, large grit and acid-etched, and no treatment (control). The proliferation and osteogenic differentiation of BMSCs on the different surface morphologies were observed with the same concentration of EMDs. To further understand the osteogenic mechanism of EMDs on BMSC sheets with different morphologies, a real-time RT-PCR and a western blot were used to detect the overall levels of osteogenic genes and osteogenic proteins. Finally, to verify the osteogenic effect of BMSC sheets stimulated by EMDs in vivo, BMSC sheets with different morphologies were implanted into the subcutaneous tissue of the back of nude mice, and the bone formation was detected by HE staining., Results: The EMDs and surface morphology in the AO group synergically increased the expression levels of osteogenic active factors (RUNX2, OSX and OCN) and enhanced the osteogenic differentiation effect of BMSCs. The in vivo experiments showed that the BMSC sheets in the AO group were rich in osteogenic active factors, and promoted the formation of ectopic bone tissue after implantation into the subcutaneous tissue of the back of nude mice., Conclusion: EMDs and AO morphology synergically enhance the secretion of bone osteogenic active factors of BMSCs and promote the formation of heterotopic bone., Competing Interests: None., (AJTR Copyright © 2023.)

Fall armyworm Spodoptera frugiperda , a native insect pest in tropical and subtropical America, has rapidly spread to most parts of China and become a major pest of corn and other crops since invading in early January 2019. As an emergency and important control measure, chemical control of S. frugiperda has the advantages of quick effect and low cost. However, long-term and large-scale use of pesticides might pollute the environment and increase pest resistance. By improving the control effect and reducing the dosage of chemical pesticides, graphene oxide (GO) is used synergistically with insecticides to increase control efficacy to achieve low-cost and sustainable management of insect pests as a new type of synergist. In this study, graphene oxide was compounded with insecticides to form nanocomposites. To clarify pest physiological responses, the laboratory toxicity of graphene oxide-insecticide nanocomposites was measured on the larvae of S. frugiperda . The results demonstrated that GO could enhance the activity of four selected pesticides: chlorantraniliprole (Chl), beta cypermethrin (Bet), methoxyhydrazide (Met) and spinetoram (Spi). Compared with pesticides alone, the toxicity of Chl-GO, Bet-GO, Met-GO and Spi-GO mixtures to the third instar larvae of S. frugiperda increased by 1.56, 1.54, 2.53 and 1.74 times, respectively. The easy preparation and higher bioactivity of GO-pesticide nanocomposites indicated their promising application potential in pest control.

To study the effect of miR-153-3p on the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) in a high glucose environment and its potential mechanism. The results showed that high glucose inhibited the osteogenic differentiation of BMSCs, and the expression of miR-153-3p increased during osteogenic differentiation. Further experiments found that in BMSCs induced by high glucose, overexpression of miR-153-3p inhibited the osteogenic differentiation of BMSCs, and the expressions of osteogenesis-related genes bone sialoprotein, Collagen I and alkaline phosphatase were down-regulated, while silencing of miR-153-3p alleviated the inhibition effect. The dual-luciferase reporter gene assay confirmed that the 3'-untranslated region (3'-UTR) of runt related transcription factor 2 (RUNX2) had a targeted binding site with miR-153-3p and a negative regulatory effect. Molecular studies further confirmed that miR-153-3p inhibited the osteogenic differentiation of BMSCs by targeting the 3'-UTR of RUNX2. In conclusion, our study found that as one key regulator of high glucose affecting the osteogenic differentiation of BMSCs, miR-153-3p may play a negative regulatory role by inhibiting the expression of RUNX2., Competing Interests: None., (AJTR Copyright © 2022.)

Digestive system diseases arise primarily through the interplay of genetic and environmental influences; there is an urgent need in elucidating the pathogenic mechanisms of these diseases and deploy personalized treatments. Traditional and long-established model systems rarely reproduce either tissue complexity or human physiology faithfully; these shortcomings underscore the need for better models. Organoids represent a promising research model, helping us gain a more profound understanding of the digestive organs; this model can also be used to provide patients with precise and individualized treatment and to build rapid in vitro test models for drug screening or gene/cell therapy, linking basic research with clinical treatment. Over the past few decades, the use of organoids has led to an advanced understanding of the composition of each digestive organ and has facilitated disease modeling, chemotherapy dose prediction, CRISPR-Cas9 genetic intervention, high-throughput drug screening, and identification of SARS-CoV-2 targets, pathogenic infection. However, the existing organoids of the digestive system mainly include the epithelial system. In order to reveal the pathogenic mechanism of digestive diseases, it is necessary to establish a completer and more physiological organoid model. Combining organoids and advanced techniques to test individualized treatments of different formulations is a promising approach that requires further exploration. This review highlights the advancements in the field of organoid technology from the perspectives of disease modeling and personalized therapy., (© 2022. The Author(s).)

Xenorhabdus nematophila HB310 secreted the insecticidal protein toxin complex (Tc). The chi60 and chi70 chitinase genes are located on the gene cluster encoding Tc toxins. To clarify the insecticidal activity of chitinases and their relationship with Tc toxins, the insecticidal activity of the chitinases was assessed on Helicoverpa armigera . Then, the chi60 and chi70 genes of X. nematophila HB310 were knocked out by the pJQ200SK suicide plasmid knockout system. The insecticidal activity of Tc toxin from the wild-type strain (WT) and mutant strains was carried out. The results demonstrate that Chi60 and Chi70 had an obvious growth inhibition effect against the second instar larvae of H. armigera with growth-inhibiting rates of 81.99% and 90.51%, respectively. Chi70 had a synergistic effect with the insecticidal toxicity of Tc toxins, but Chi60 had no synergistic effect with Tc toxins. After feeding Chi60 and Chi70, the peritrophic membrane of H. armigera became inelastic, was easily broken and leaked blue dextran. The Δ chi60 , Δ chi70 and Δ chi60 - chi70 mutant strains were successfully screened. The toxicity of Tc toxins from the WT, Δ chi60 , Δ chi70 and Δ chi60 - chi70 was 196.11 μg/mL, 757.25 μg/mL, 885.74 μg/mL and 20,049.83 μg/mL, respectively. The insecticidal activity of Tc toxins from Δ chi60 and Δ chi70 was 3.861 and 4.517 times lower than that of Tc toxins from the WT, respectively, while the insecticidal activity of Tc toxins from the Δ chi60 - chi70 mutant strain almost disappeared. These results indicate that the presence of chi60 and chi70 is indispensable for the toxicity of Tc toxins.

Introduction: The influence of enamel matrix derivative (EMD) on proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) was explored in high glucose (HG) microenvironment with interaction of Wnt/β-catenin pathway., Materials and Methods: Extraction of BMSCs from Sprague-Dawley rats, culture, and identification were manifested. The cells were treated with different concentration of EMD in HG to figure out the most available concentration for proliferation and osteogenic differentiation. Then, observation of cell growth curve and cell cycle changes, and detection of Osterix, runt-related transcription factor 2 (Runx2), COL-I, early osteogenic indexes, Calcium salt deposition, and β-catenin protein in Wnt/β-catenin pathway were assured. After adding Wnt/β-catenin pathway inhibitor (XAV-939) in the cells with osteogenesis induction, detection of binding of β-catenin to Osterix was clarified., Results: Via identification BMSCs cultured in vitro was qualified. Different concentrations of EMD could accelerate cell proliferation in HG and osteogenesis induction, and 75 μg/mL EMD had the best effect. The HG augmented BMSCs proliferation and the propidium iodide index of flow cytometry cycle was elevated in HG, which were strengthened via the EMD. After BMSCs' osteogenesis induction, Osterix, Runx2, CoL-1, early osteogenic indexes, and calcium salt deposition were reduced, but elevated via EMD. β-Catenin was the lowest in the HG, but elevated after EMD. After addition of XAV-939, reduction of β-catenin and the downstream (Osterix and Runx2) were manifested. Detection of binding protein bands was in β-catenin and Osterix of the HG after EMD treatment., Conclusion: EMD may facilitate the osteogenic differentiation of BMSCs via activating the Wnt/β-catenin pathway in HG., (© 2022. The Japanese Society Bone and Mineral Research.)

The main aim of this research work was to develop and evaluate a drug delivery system with compression coating technology to control drug release at a constant rate. The compression coated tablets (CCTs) consist of the hydrophilic matrix core and the hydrophobic waxy coating. The presence of hydrophobic waxy coating could provide sufficient time for hydration of the core to prevent initial burst release. The mechanism research revealed that erosion was the main way of drug release and the releasing area was constant during the entire release process because the core tablet was located in the cup-shaped coating after one side cover was dropped at the lag time. This made the release behavior exhibit zero-order kinetics ( R 2 >0.99). The coating rupture strength and the core swelling force at the lag time influenced erosion rate thus affecting release rate. Different solubility of drugs (propranolol hydrochloride, melatonin, and nifedipine) was selected as model drugs and the properties of the prepared CCTs in terms of formulations and in vitro release were evaluated. The release rate was independent of solubility, medium pH, and osmotic pressure. This zero-order controlled system could be applied to both controlled drug delivery and chrono pharmaceutical drug delivery.

Insect glutathione-S-transferases (GSTs) play essential roles in metabolizing endogenous and exogenous compounds. GSTs that are uniquely expressed in antennae are assumed to function as scavengers of pheromones and host volatiles in the odorant detection system. Based on this assumption, antennae-specific GSTs have been identified and functionally characterized in increasing number of insect species. In the present study, 17 putative GSTs were identified from the antennal transcriptomic dataset of the Indian meal moth, Plodia interpunctella , a severe stored-grain pest worldwide. Among the GSTs, only PiGSTd1 is antennae-specific according to both Fragments Per Kilobase Million (FPKM) and quantitative real-time PCR (qRT-PCR) analysis. Sequence analysis revealed that PiGSTd1 has a similar identity as many delta GSTs from other moths. Enzyme kinetic assays using 1-chloro-2,4-dinitrobenzene (CDNB) as substrates showed that the recombinant PiGSTd1 gave a K m of 0.2292 ± 0.01805 mM and a V max of 14.02 ± 0.2545 μmol·mg -1 ·min -1 under the optimal catalytic conditions (35°C and pH = 7.5). Further analysis revealed that the recombinant PiGSTd1 could efficiently degrade the sex pheromone component Z9-12:Ac (75.63 ± 5.52%), as well as aldehyde volatiles, including hexanal (89.10 ± 2.21%), heptanal (63.19 ± 5.36%), ( E )-2-octenal (73.58 ± 3.92%), ( E )-2-nonenal (75.81 ± 1.90%), and ( E )-2-decenal (61.13 ± 5.24%). Taken together, our findings suggest that PiGSTd1 may play essential roles in degrading and inactivating a variety of odorants, especially sex pheromones and host volatiles of P. interpunctella ., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Liu, Tang, Wang, Shi, Yin and Li.)

Serotonin receptor 6 (5-HT 6 R), a typical G protein-coupled receptor (GPCR) mainly expressed in the neurogenic area with constitutive activity, is of particular interest as a promising target for emotional impairment. Here, we found that 5-HT 6 R was highly expressed in human NSCs and activation of the receptor promoted self-renewal of human NSCs, and thus induced the expansion and folding of human cerebral organoids; dysfunction of receptor or inhibition of its constitutive activity resulted in the premature differentiation of NSCs, which ultimately depleted the NSC pool. The following mechanistic study revealed that EPAC-CREB signaling was involved in 5-HT 6 R regulation. Furthermore, we showed that mice with genetic deletion of 5-HT 6 R or knockin A268R mutant presented depression-like behaviors and impaired hippocampal neurogenesis for progressive decrease of the NSC pool. Thus, this study indicates that the modulation of 5-HT 6 R and its constitutive activity may provide a therapeutic alternative to alleviate depression., (Copyright © 2020 The Authors. Published by Elsevier Inc. All rights reserved.)

The intricate balance of neural stem cell (NSC) amplification and neurogenesis is central to nervous system development. Dopamine D1 receptor (DRD1) is a typical G protein-coupled receptor (GPCR) mainly expressed in neurogenic area, with high constitutive activity. The receptor appears in the embryonic period before the formation of mature synaptic contacts, which indicates that dopamine receptor and its constitutive activity play crucial roles in the embryonic brain development. Here, we found that DRD1 was enriched in human NSCs. Inhibition of the receptor activity by its inverse agonists promoted human NSCs proliferation and impeded its differentiation. These results were also mimicked by genetic knockdown of DRD1, which also blocked the effects of inverse agonists, suggesting a receptor-dependent manner. More interestingly, knock-in A229T mutant with reduced DRD1 constitutive activity by CRISPR-Cas9 genome editing technology resulted into increased endogenous human NSCs proliferation. These results were well reproduced in human cerebral organoids, and inhibition of the DRD1 constitutive activity by its inverse agonists induced the expansion and folding of human cerebral organoids. The anatomic analysis uncovered that decreasing the constitutive activity of DRD1 by its inverse agonists promoted the NSCs proliferation and maintenance that led to hindered cortical neurogenesis. Further mechanistic studies revealed that the PKC-CBP pathway was involved in the regulation by DRD1. Thus, our findings indicate that the constitutive activity of DRD1 and possibly other GPCRs plays an important role in the development of human nervous system., (©2020 The Authors. Stem Cells published by Wiley Periodicals, Inc. on behalf of AlphaMed Press 2020.)

PirAB toxin was initially found in the Photorhabdus luminescens TT01 strain and is a demonstrated binary toxin with high insecticidal activity. In this paper, we co-expressed the pirAB gene of Xenorhabdus nematophila HB310 in a prokaryotic expression system, and we found that the PirAB protein showed high hemocoel insecticidal activity against Galleria mellonella, Helicoverpa armigera and Spodoptera exigua. LD 50 values were 1.562, 2.003 and 2.17 μg/larvae for G. mellonella, H. armigera, and S. exigua, respectively (p > 0.05). Additionally, PirAB-interaction proteins were identified from G. mellonella by 6 × His Protein Pulldown combined with liquid chromatography-tandem mass spectrometry (LC-MS/MS). Of which, arylphorin of G. mellonella showed the highest matching rate. A protein domain conservative structure analysis indicated that arylphorin has three domains including Hemocyanin-N, Hemocyanin-M, and Hemocyanin-C. Among these protein domains, Hemocyanin-C has immune and recognition functions. Further, Hemocyanin-C domain of arylphorin was identified to interact with PirA but not PirB by Yeast two-hybrid system. These findings reveal, for the first time, new host protein interacting with PirAB. The identification of interaction protein may serve as the foundation for further study on the function and insecticidal mechanism of this binary toxin from Xenorhabdus.

Rationale: Cellular angiofibroma is a rare benign mesenchymal tumor which mostly occurs in the superficial soft tissues of the genital region. Occurrence in the head and neck region is extremely rare. To our knowledge, this is the first case of cellular angiofibroma arising in the hypopharynx., Patient's Concerns: A 54-year-old male complained of a mass moving through his throat for 1 month. A tumor was found in the right lateral wall of the pharyngeal at the level of the epiglottis by laryngoscope. Magnetic resonance imaging confirmed the presence of a mass in the right lateral pharyngeal area. A benign tumor was suspected based on the clinical symptoms and imaging findings., Diagnosis: A supporting laryngoscope was performed under general anesthesia and the lesion was resected. Immunohistochemical analysis revealed cellular angiofibroma., Interventions: The patient underwent surgical excision of the lesion., Outcomes: Thus far, no recurrence has been observed 6 months after excision., Lessons: Cellular angiofibroma located in the lateral pharyngeal is rare; however, immunohistochemical staining is helpful for its diagnosis. Treatment is relatively simple and requires local excision and follow-up.

This retrospective study evaluated operative outcomes when using a supraclavicular artery island flap (SAIF) combined with a sternohyoid muscle flap (SHMF) to reconstruct defects after hypopharyngeal carcinoma resection. Reconstructive surgery for hypopharyngeal and laryngeal defects was performed with the SAIF + SHME combination in 6 patients during 2016 to 2018. Within 14 to 16 days after the surgery, all 6 patients could ingest food and block the tube (avoiding aspiration), with no pharyngeal fistulas. They then underwent irradiation up to a total of 60.5 Gy during the 4 weeks postoperatively. All 6 flaps survived, and there were no donor-site complications except minor dehiscence in 1 patient. Thus, the SAIF + SHMF combination can be used to reconstruct hypopharyngeal and laryngeal defects after hemi-laryngectomy in patients with hypopharyngeal carcinoma involving the unilateral larynx. This technique effectively preserved the swallowing function and phonation of the patients, thereby improving their quality of life.

Background: Genome sequence analysis (GenBank access No.: FN667742.1) shows that Xenorhabdus nematophila ATCC19061 contains one gene (Xn-cbp) encoding chitin binding protein (Xn-CBP)., Objective: The present work aims to clarify the characteristics and function of Xn-CBP from X. nematophila HB310., Methods: In this study, the Xn-cbp gene was cloned and expressed in Escherichia coli BL21 (DE3). Substrate binding assays were performed to explain the ability of Xn-CBP combined with the polysaccharide. The insecticidal toxicity of Xn-CBP against the second-instar larvae of Helicoverpa armigera was determined by feeding method. Besides, the antifungal activity of Xn-CBP against Coniothyrium diplodiella, Verticillium dahlia, and Fusarium oxysporum was tested by spore germination assay and hyphal extension assay., Results: Xn-CBP encoded 199 amino acids with a calculated mass of 28 kDa, which contained a signal peptide and a chitin binding domain. The Bmax and Kd values of Xn-CBP to colloidal chitin were 2.46 and 4.08, respectively. Xn-CBP had insecticidal activity against the H. armigera with a growth inhibition rate of 84.08%. Xn-CBP had the highest spore germination inhibitory effect on C. diplodiella with the inhibition rate of 83.11%. The hyphal growth inhibition rate of Xn-CBP to F. oxysporum, 41.52%, was higher than the other two fungi., Conclusion: The Xn-CBP had the highest binding ability to colloidal chitin and it showed insecticidal activity and antifungal activity. The present study laid a foundation for further exploitation and utilization of X. nematophila., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

Xenorhabdus nematophila HB310 secreted the insecticidal protein toxin complex. Two chitinase genes, chi60 and chi70, were found in X. nematophila toxin complex locus. In order to clarify the function of two chitinases, chi60 and chi70 genes were cloned and expressed in Escherichia coli Transetta (DE3). As a result, we found that the Chi60 and Chi70 belonged to glycoside hydrolases (GH) family 18 with a molecular mass of 65 kDa and 78 kDa, respectively. When colloidal chitin was treated as the substrate, Chi60 and Chi70 were proved to have the highest enzymatic activity at pH 6.0 and 50 °C. Chi60 and Chi70 had obvious growth inhibition effect against the second larvae of Helicoverpa armigera with growth inhibiting rate of 81.99% and 90.51%. Chi70 had synergistic effect with the insecticidal toxicity of Bt Cry 1Ac, but the Chi60 had no synergistic effect with Bt Cry 1Ac. Chi60 and Chi70 showed antifungal activity against Alternaria brassicicola, Verticillium dahliae and Coniothyrium diplodiella. The results increased our understanding of the chitinases produced by X. nematophila and laid a foundation for further studies on the mechanism of the chitinases.

Rationale: Adenoid cystic carcinomas (ACCs) are malignant tumors and occur in the major and the minor salivary glands. ACCs are rare in the larynx., Patient Concerns: A 55-year-old female patient who presented with a 12 months history of paraesthesia pharynges and hoarseness for 4 months. Laryngoscopy showed an endophytic lesion in the right supglottic area with no laryngeal stenosis. And magnetic resonance imaging (MRI) confirmed the presence of a submucosal mass in the supglottic area. Supporting laryngoscope was performed under general anesthesia and a lesion biopsy obtained., Diagnoses: The lesion was sent for frozen-section biopsy, which shows the lesion was the epithelial-derived tumors of salivary gland, but it was impossible to identify the relationship between the tumor and the surrounding normal tissue, and to judge the nature of the tumor further. So, the lesion was sent for the second frozen-section biopsy, an ACC of the larynx was confirmed and margins were negative., Interventions: The patient underwent partial laryngectomy., Outcomes: The patient was doing well except hoarseness during the follow-up period with no evidence of disease recurrence or metastasis for 17 months post-operatively., Lessons: Laryngeal ACC is a rare entity. When the nature of lesion cannot be confirmed, multiple biopsies may be required for confirm the diagnosis of pathology. It not only reduces patient's waiting time for surgery, but also define the diagnosis and surgical removal of lesion through a single anesthesia.

Objective: Adherence of pathogen to nasal mucosa and colonization is the first step of bacterial biofilm(BBF) formation in patients with chronic rhinosinusitis (CRS).Terminal sialic acids presenting on cell surface are potential targets for bacterial binding, thus may partly contribute to the pathogenesis of CRS. However, little has been published in this respect, the purpose of our study aimed to investigate the expression of sialic acids on the nasal mucosa in CRS patients and its possible effect on BBF formation., Methods: Sinus mucosa were harvested from CRS patients undergoing endoscopic surgery. The positive of BBF formation were detected by scanning electronic microscopy (SEM) and the expression of Neu5Acα2,3Gal(α2,3-linked sialic acid) and Neu5Acα2,6Gal(α2,6-linked sialic acid) on nasal mucosa were determined by fluorescent-immunohistochemical staining (F-IHC) with MAL-II and SNA respectively. A semi-quantitative scoring system was used to assess their different expression between CRS group and the control, as well as BBF positive and negative group., Results: Expression of Neu5Acα2,3Gal and Neu5Acα2,6Gal were both detected in the epithelium and submucosal glands of all 40 CRS patients and 23 controls, they were significantly up-regulated in CRS group(p < 0.05). Among 24 CRS patients, typical BBF formation were identified in 13 cases while the other 11 were regarded as negative, Between the subgroup of BBF(+) and BBF(-), both of Neu5Acα2,3Gal and Neu5Acα2,6Gal had a trend of increasing in BBF(+) group, however, the increased expression of Neu5Acα2,3Gal was statistical significance (4.77 ± 0.90 versus 3.45 ± 1.40; p = 0.0282), whereas the difference of Neu5Acα2,6Gal was insignificant(4.15 ± 1.27 versus 3.55 ± 1.59; p = 0.4281)., Conclusion: Expression of MAL-II binding (most probable Neu5Acα2,3Gal) and SNA binding (Neu5Acα2,6Gal) were up-regulated in inflamed nasal mucosa, and the increased expression of them may contribute to bacterial biofilm formation which deserved a further investigation., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

β-Arrestins (β-arrestin-1 and -2) are multifunctional proteins that play important roles in the regulation of inflammation and cell survival that need to be tightly controlled; however, the mechanism that underlies their gene expression is largely unclear. Here, we demonstrate that β-arrestin-1 is a transcriptional target of NF-κB. mRNA and protein levels of β-arrestin-1 were up-regulated by NF-κB inducers. Inhibition of NF-κB prevented the up-regulation of β-arrestin-1 mRNA, whereas activation of NF-κB led to increased β-arrestin-1 expression. β-Arrestin-1 promoter activity was consistently enhanced upon NF-κB activation as a result of the presence of a highly conserved κB site. β-Arrestin-1, in turn, suppressed the transcriptional activity of NF-κB by interfering with the interaction between p65 and p50. β-Arrestin-1-deficient mice displayed reduced TNF-α-induced cell death and increased expression of antiapoptotic genes. Reintroduction of β-arrestin-1, but not its mutant, which is unable to interfere with the p65-p50 interaction, into β-arrestin-deficient mouse embryonic fibroblasts partially restored sensitivity to TNF-α-induced cell death. These findings reveal NF-κB and β-arrestin-1 to be key components of a negative feedback circuit that is necessary to regulate cell death.-Li, J., Guo, A., Wang, Q., Li, Y., Zhao, J., Lu, J., Pei, G. NF-κB directly regulates β-arrestin-1 expression and forms a negative feedback circuit in TNF-α-induced cell death.

The influence of meteorological determinants on tuberculosis (TB) incidence remains severely under-discussed, especially through the perspective of time series analysis. In the current study, we used a distributed lag nonlinear model (DLNM) to analyze a 10-year series of consecutive surveillance data. We found that, after effectively controlling for autocorrelation, the changes in meteorological factors related to temperature, humidity, wind and sunshine were significantly associated with subsequent fluctuations in TB incidence: average temperature was inversely associated with TB incidence at a lag period of 2 months; total precipitation and minimum relative humidity were also inversely associated with TB incidence at lag periods of 3 and 4 months, respectively; average wind velocity and total sunshine hours exhibited an instant rather than lagged influence on TB incidence. Our study results suggest that preceding meteorological factors may have a noticeable effect on future TB incidence; informed prevention and preparedness measures for TB can therefore be constructed on the basis of meteorological variations.

The aim of this study is to evaluate the efficacy of endoscopic treatment for maxillary inverted papilloma (IP) through partial medial maxillectomy with an inferior turbinate reversing approach. A retrospective analysis of patients treated in our institution for maxillary sinus IP between July 2011 and August 2015 was performed. Demographics, operative technique, characteristics of tumors, complications, postoperative follow-up, and recurrence were evaluated. Twenty-two patients were enrolled in the study. All tumor attachments were identified intraoperatively. Adequate visualization was obtained following our approach. All inferior turbinate and nasolacrimal ducts were preserved. The median follow-up time was 41 months. One recurrence occurred at the follow-up time of 27 months. Postoperative hemorrhage and numbness at the ipsilateral frontal teeth were reported in two and one patients, respectively. Endoscopic surgery through partial medial maxillectomy using an inferior turbinate reversing approach provides full access to the maxillary sinus and preserves the inferior turbinate and nasolacrimal duct.

PirAB (Photorhabdus insect-related proteins, PirAB) toxin was initially found in the Photorhabdus luminescens TT01 strain and has been shown to be a binary toxin with high insecticidal activity. Based on GenBank data, this gene was also found in the Xenorhabdus nematophila genome sequence. The predicted amino acid sequence of pirA and pirB in the genome of X. nematophila showed 51% and 50% identity with those gene sequences from P. luminescens. The purpose of this experiment is to identify the relevant information for this toxin gene in X. nematophila. The pirA, pirB and pirAB genes of X. nematophila HB310 were cloned and expressed in Escherichia coli BL21 (DE3) using the pET-28a vector. A PirAB-fusion protein (PirAB-F) was constructed by linking the pirA and pirB genes with the flexible linker (Gly) 4 DNA encoding sequence and then efficiently expressed in E. coli. The hemocoel and oral insecticidal activities of the recombinant proteins were analyzed against the larvae of Galleria mellonella. The results show that PirA/B alone, PirA/B mixture, co-expressed PirAB protein, and PirAB-F all had no oral insecticidal activity against the second-instar larvae of G. mellonella. Only PirA/B mixture and co-expressed PirAB protein had hemocoel insecticidal activity against G. mellonella fifth-instar larvae, with an LD 50 of 2.718μg/larva or 1.566μg/larva, respectively. Therefore, we confirmed that PirAB protein of X. nematophila HB310 is a binary insecticidal toxin. The successful expression and purification of PirAB laid a foundation for further studies on the function, insecticidal mechanism and expression regulation of the binary toxin., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Alzheimer's disease (AD) is a major and devastating neurodegenerative disease, and the amyloid-β (Aβ) hypothesis is still the central theory for AD pathogenesis. Meanwhile, another major mental illness, depression, is one of the risk factors for AD. From a high-throughput screening (HTS), amoxapine, a typical secondary amine tricyclic antidepressant (TCA), was identified to reduce Aβ production. A follow-up investigation on antidepressants showed that most of the TCAs harbour similar activity. Previous studies have indicated that TCAs improve cognitive function in AD mouse models as well as in preliminary clinical data; however, the underlying mechanism is controversial, and the effect on Aβ is elusive. Thus, we developed a secondary screening to determine the molecular target of amoxapine, and serotonin receptor 6 (HTR6) was identified. Knockdown of HTR6 reduced the amoxapine's effect, while the HTR6 antagonist SB258585 mimicked the activity of amoxapine. Further mechanistic study showed that amoxapine and SB258585 reduced Aβ generation through multiple HTR6-mediated targets, including β-arrestin2 and CDK5. Taken together, our study suggests that amoxapine, though no longer a first-line drug for the treatment of depression, may be beneficial for AD and further structural modification of TCAs may lead to desirable therapeutic agents to treat both AD and depression.

Although levodopa is the first-line medication for the treatment of Parkinson's disease (PD) showing unsurpassable efficiency, its chronic use causes dyskinesia. Accordingly, dopamine agonists are increasingly employed as monotherapy or in combination with levodopa to reduce the risk of motor complications. It is well recognized that patients with PD often exhibit cognitive deficits. However, clinical and animal studies assessing the effects of dopaminergic medications on cognition are controversial. Amyloid-β (Aβ) is one of the major hallmarks of Alzheimer's disease (AD), leading to progressive memory loss and cognitive deficit. Interestingly, the abnormal accumulation of Aβ is also detected in PD patients with cognitive deficits. Evidence indicated that levodopa induced a mild increase of Aβ plaque number and size in the brain of AD mouse. However, the underlying mechanism is unclear. Here we present that both levodopa and piribedil enhance the generation of Aβ and the activity of γ-secretase in human neuronal cells and primary neurons isolated from AD mouse. This effect was reduced by either the antagonism or the knockdown of dopamine D2 receptor (D2R). We further showed that in the cells expressing β-arrestin 2-biased D2R mutant, piribedil promoted cellular Aβ production to the extent comparable to the wild-type D2R whereas this activity was absent in those with G protein-biased D2R mutant. Moreover, the knockdown of β-arrestin 2 attenuated the increases of Aβ generation and γ-secretase activity mediated by levodopa or piribedil. Thus, our study suggests that targeting D2R-mediated β-arrestin function may have potential risk in the modulation of Aβ pathology.

MicroRNAs (miRNAs) have been widely demonstrated to play fundamental roles in gene regulation in most eukaryotes. To date, there has been no study describing the miRNA composition in genetically modified organisms (GMOs). In this study, small RNAs from dry seeds of two GM soybean lines and their parental cultivars were investigated using deep sequencing technology and bioinformatic approaches. As a result, several differentially expressed gma-miRNAs were found between the GM and non-GM soybeans. Meanwhile, more differentially expressed gma-miRNAs were identified between distantly relatednon-GM soybeans, indicating that the miRNA components of soybean seeds varied among different soybean lines, including the GM and non-GM soybeans, and the extent of difference might be related to their genetic relationship. Additionally, fourteen novel gma-miRNA candidates were predicted in soybean seeds including a potential bidirectionally transcribed miRNA family with two genomic loci (gma-miR-N1). Our findings firstly provided useful data for miRNA composition in edible GM crops and also provided valuable information for soybean miRNA research.

Neuronal conversion from human fibroblasts can be induced by lineage-specific transcription factors; however, the introduction of ectopic genes limits the therapeutic applications of such induced neurons (iNs). Here, we report that human fibroblasts can be directly converted into neuronal cells by a chemical cocktail of seven small molecules, bypassing a neural progenitor stage. These human chemical-induced neuronal cells (hciNs) resembled hiPSC-derived neurons and human iNs (hiNs) with respect to morphology, gene expression profiles, and electrophysiological properties. This approach was further applied to generate hciNs from familial Alzheimer's disease patients. Taken together, our transgene-free and chemical-only approach for direct reprogramming of human fibroblasts into neurons provides an alternative strategy for modeling neurological diseases and for regenerative medicine., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Objective: To investigate the association of mutations in the hepatitis B virus (HBV) X gene (HBX, encoding the HBx protein) and development of hepatocellular carcinoma (HCC)., Methods: Forty-four patients with HBV-related HCC participated in the study, along with 76 patients with chronic HBV infection who assessed as controls. All patients had serum HBV DNA levels that were higher than 10(3) copies/ml. Extracted HBV DNA was subjected to nested PCR to amplify the HBX gene, followed by direct sequencing. All sequencing data were compared to the consensus HBV sequence to identify mutations. The sequencing data were analyzed by Chromas and SeqMan software., Results: Mutations of G1467C, G/C1479A, C1485T and C1653T in the X region were found, but did not show any significant difference in occurrence between the HCC group and the chronic HBV infection group (P>0.05). The T1674C mutation in the X region, however, occurred more frequently in the HCC group (29.27% vs.6.67%, P<0.05). Prevalence of the T1753C mutation and the A1762T/G1764A double mutation in the BCP region was significantly higher in the HCC group than in the chronic HBV infection group (P<0.05) and in the group of patients with hepatitis B e antigen (HBeAg)-negative status compared to the patients with HBeAg-positive status (P<0.05)., Conclusion: Incidence of the T1674C mutation in the X region and of the T1753C mutation and the A1762T/G1764A double mutation in the BCP region was higher for patients with HBV-related HCC; the T1753C mutation and the A1762T/G1764A double mutation may inhibit the formation of HBeAg.

Objectives: To determine the outcomes of coblation assisted transnasal endoscopic resection of nasopharyngeal cyst., Method: Retrospective chart review outcomes in 12 patients who underwent endoscopic resect cysts of the nasopharynx at our department between 2001 and 2010. Twelve patient, aged 28 to 71 years, with cysts of the nasopharynx. The outcome variables of complications and the rate of recurrence were analyzed, respectively., Results: In 12 cases, retention cysts in 2 cases, branchial cyst in 1 case, adenoid middle fossa cyst infection in 6 cases, Tornwaldt's cyst in 3 cases. The use of the coblation device was associated with a significant decrease in blood loss. There were no postoperative complications, and the overall follow-up period was 2-7 years and shows no signs of recurrence., Conclusions: We describe transnasal endoscopic procedures to resect cysts of the nasopharynx. We found that radiofrequency coblation is a useful and safe tool associated with minimal blood loss in the resection of these cysts. In our experience, it has been a highly successful, safe, and effective procedure.

Basal cell adenoma is an uncommon benign salivary gland neoplasm, presenting isomorphic basaloid cells with a prominent basal cell layer. Basal cell adenoma arising from the nasal septum is exceptionally rare. Reports on positron emission tomography with 2-deoxy-2-fluorine-18-fluoro-D-glucose (18FDG-PET) imaging for basal cell adenoma are limited. Here, we present the case of a 49-year-old man who had the symptoms of intermittent repeated bleeding from the left nose for half a year. 18FDG-PET scanning showed increased accumulation of 18FDG with its characteristic benign pathology has a potential to malignancy. After removal of the mass, the patient became symptom free. Pathology showed basal cell adenoma. The evidence of active and growing cells was present in the specimen.

Primary chondroid tumors of the larynx represent less than 1% of all laryngeal tumors. Most of them are chondromas and they often involve to the cricoid cartilage. They are characterized by a low tendency to metastatic diffusion (low grade). The treatment of choice is surgery, which may be endoscopic or "open partial surgery", if extension of the cancer is limited. Prognosis is generally good. In this report, two cases of low grade chondrosarcoma of the larynx are presented, one was treated surgically with cricoidectomy and partial laryngectomy, and another was treated surgically with hemicricoidectomy. Laryngoscopy reveals tumefaction of the larynx, covered by intact mucosa. Computerized tomography imaging with contrast and magnetic resonance imaging defines not only coarse calcifications, pathognomonic of chondromatous neoformations but also the relationship of the neoformation with the surrounding tissues. Treatment is essentially surgical, given the importance of preserving the larynx to patients' quality of life, the only risk is recurrence, which is treated by a second surgery.

Typical carcinoid tumor of the larynx is an extremely rare lesion which arises from neuroendocrine cells scanning in the laryngeal mucosa or submucosal glands. Conventionally, it is a well-differentiated neuroendocrine carcinoma, conservative surgery represents the treatment of choice, and the patient usually has a good prognosis with rarely recurrence and metastasis. In this report, we present a case of typical laryngeal carcinoid tumor with recurrence and lymph node metastasis. The patient was a 58-year-old man, complaints of intermittent burning pain in his both ears for 2 years, and for the recent one month the pain turn to continuous accompanied with a mild discomfort in the throat, he had neither hoarseness, dysphagia, nor any otorrhea and hearing loss. The patient was scheduled to undergo a tracheotomy and then a biopsy under supporting laryngoscopy. During the operation, the frozen section diagnosis from the first and the second time both indicated that the biopsy specimens originated from musculo-epithelia, it couldn't be differed from malignant to benign. So the mass was simply removed under supporting laryngoscopy. The histopathology from paraffin sections revealed typical carcinoid of the larynx and the second procedure consisted of supraglottic laryngectomy with clear margins, the otalgia resolved and the patient had no difficulty with phonation or swallowing. After 1 year follow-up, the patient was found a mass on his right neck with symptom free, B-ultrasonography indicated several enlargement lymph nodes with some merged on both sides of the neck, the patient was scheduled to undergo a "total-laryngectomy with radical neck dissection on the left side and an elective neck dissection on the right side". The specimens were positive for cytokeratin, chromogranin A and synaptophysin, a final diagnosis of typical carcinoid was made for the recurrence lesion and the metastasis of the lymph nodes. Though the post-operative recovery was uneventful, the prognosis was not good, the patient died six months later. Here, we review the pertinent references on this subject, and discuss the main managements for typical carcinoid tumor of the larynx.

Bacillus thuringiensis Cry7Ab3 toxin has insecticidal activity against larvae of Henosepilachna vigintioctomaculata. Cry7Ab3 toxin is solubilized under alkaline condition and activated by proteases within the larval gut. In order to assess the functions of the N- and C-terminal regions, several N- and C-terminal truncated forms of Cry7Ab3 were constructed. It was determined that amino acid removal at the N-terminal, which disrupt the α-helico structure, resulted in the inactivation of the protein. The deletion of 512 amino acids from the C-terminus reduced the toxicity. However, the deletion of 481 amino acids from the C-terminus resulted in the highest activity. These findings directly demonstrated the critical roles of N- and C-terminal amino acids on the toxicity of Cry7Ab3., (© 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Amyloidosis is a spectrum of diseases characterized by abnormal extracellular accumulation of proteinaceous material; its precise etiology still remains unclear. It may affect multiple organs, of which the commonest sites are larynx, bronchus and kidney. Laryngeal amyloidosis is usually a localized phenomenon rarely associated with systemic involvement, here we report a case of laryngeal amyloidosis with tracheobronchial involved. The patient was 31-years old. He had a history of open surgical operation for laryngeal amyloidosis in the left ventricle 4-years ago. This time he was suffered by amyloid deposition in his right ventricle, the lesion was excised on staged laryngoscopy under general anesthesia. After 20-months follow-up, the post-operative recovery was wonderful. The bronchoscopy examination and computerized tomography scan for chest demonstrated he also had right main bronchus amyloidosis. Being of no dyspnea, he was unwilling to accept an operation on bronchus. Though amyloidosis is a benign lesion, up to date, there are no curable treatments for such a disease, for severe cases, it may be fatal as a result of airway obstruction or respiratory failure. Here we review the pertinent references on this subject, and discuss the main managements for amyloidosis on larynx and bronchus.

Objective: The nasal cavity is an uncommon site for metastasis to develop and thus metastases arising from breast cancer are rarely observed. We report a case of a 61-year-old female with two-year history of breast cancer who presented with a nasal cavity that was diagnosed as metastatic breast carcinoma by histopathological analysis of the nasal cavity specimen., Methods: We reviewed the clinical records of the patient and the appropriate world literature., Results: The patient had been diagnosed with breast cancer two years before. Her present complain was oculus dexter visual acuity decreased. CT and MRI scan revealed a palpable mass on the right nasal cavity. PET/CT demonstrated no additional uptake at the level of other organ. ER and PR demonstrated a similar expression pattern in primary breast carcinoma and nasal cavity lesions. As further treatment she received systemic palliative chemotherapy in addition to intravenous treatment with bisphosphonates, and a total dose of 36 Gy of X-ray (3 Gy per day, 12 fractions) was given to the local site of the right nasal cavity., Conclusion: In patient with a previous history of breast cancer who complains even of ophthalmologic symptoms such as visual acuity decreased, it is important to consider nasal cavity metastatic disease. 18FDG-PET/CT is useful to rule out the presence of other organ metastasis. Histopathological analysis may aid the diagnosis. The establishment of treatment strategies based on a comprehensive understanding of both etiology and pathophysiology is needed for rare cases such as this.

Bt WZ-9 strain, containing a single Cry7Ab3 toxin, had effective insecticidal activity against larvae of Henosepilachna vigintioctomaculata. By incubation with larvae midgut homogenate and trypsin in vitro, 130 kDa Cry7Ab3 protoxin was degraded into the ∼75 kDa proteinase-resistant fragments. In vivo analysis, 130 kDa Cry7Ab3 protoxin was also processed into ∼75 kDa fragment. Histopathological observations indicated that Cry7Ab3 ingestion by H. vigintioctomaculata larvae causes acceleration in the blebbing of the midgut epithelium cells into the gut lumen and eventual lysis of the epithelium cells resulting in larval death. A ligand blotting experiment demonstrated that Cry7Ab3 toxin bound a 220 kDa BBMV protein. This receptor protein was identified as cadherin by matrix assisted laser desorption-time of flight-mass spectrometry (MALDI-TOF-MS). The cadherin protein may be the receptor of Cry7Ab3. The data obtained may contribute to a better understanding of the mechanism of Cry7Ab3 toxin against H. vigintioctomaculata larvae., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Objective: To study the methods of the surgical treatment for the nasal septum mass., Method: Two hundred and fifty-three cases of nasal septum mass were treated by surgery in our department from March 1983 to August 2007. Their clinical data were retrospectively analyzed., Result: After follow-up, most cases had no postoperative recurrence of nasal obstruction and epistaxis., Conclusion: If the diagnosis was confirmed, the effect of the surgical treatment for the nasal septum benign tumor is satisfactory. Follow-up should be done in cases with malignant tendency and malignant tumors.

Objective: Xenorhabdus nematophila is an insect pathogen bacterium symbiotically associated with entomopathogenic nematode. The bacteria produce a number of toxins to overcome immune response of insect hosts and kill their hosts. We purified a novel haemocoel insecticidal protein from X. nematophila HB310, cloned and analysed gene sequence of this novel protein., Methods: We isolated and purified the insecticidal protein by methods of salting out and native-PAGE from the intracellular proteins of X. nematophila HB310. We tested the virulence of the protein by direct injection into fifth-instar Galleria mellonella larvae. The protein was identified by western blotting. The gene of insecticidal protein was cloned by PCR and analyzed in GenBank., Results: We purified a novel haemocoel insecticidal protein that was named as Tp40. The injectable hemocoelic potency (LD50) of Tp40 was 68.54 ng/larva against fifth-instar G. mellonella larvae. The SDS-PAGE spectrum of Tp40 only showed a single - 42kDa band. Western blotting with an antibody that was highly specific to the known Txp40 indicated that Tp40 was homologous to the known Txp40 and only existed inside cells. The nucleotide sequences of tp40 gene have been deposited in GenBank (accession number GenBank: EU095326). The size of the open reading frame of tp40 was 1107bp, encoding a peptide of 368 amino acid residues, with a theoretical molecular weight 41.5 kDa and an isoelectricpoint (IP) 8.66. The Tp40 shared 85%-99% homology of nucleotide sequences and 70%-99% amino acids with those of 13 group strains., Conclusion: Tp40 is high injectable virulent for G. mellonella larvae and its gene/protein sequence is highly conserved, which play a key role during the bacterium-nematode killing host insects process.

Objective: The aim of this study was to determine the biological significance of glucose transporter (Glut)-1 and Glut-3 expression in head and neck carcinoma (HNC)., Methods: We detected expression of Glut-1 and -3 in 38 HNCs and analyzed the relationship between increased expression and the biological behavior of HNCs., Results: The gene expression levels of Glut-1 and -3 in HNCs were significantly higher than those in adjacent cancer tissues or in normal tissues. The GLUT-1 gene level was correlated with the lymph node metastasis and clinical stage of 38 HNCs, and the GLUT-3 gene level was correlated with the lymph node metastasis of 38 HNCs. Of the 38 cases, 30 showed positive expression of Glut-1 protein. The Glut-1 protein expression level was related to the lymph node metastasis and clinical stage of 38 HNCs. However, there was no case that expressed Glut-3 protein. The high expression of Glut-1 gene and protein was associated with poor survival in the HNCs. GLUT-3 gene expression was not associated with the prognosis of HNCs., Conclusions: GLUT-1 gene expression level and protein expression were correlated with lymph node metastasis, poor survival and clinical stage of HNCs. The GLUT-3 gene expression level was high in HNCs, and its expression was associated with an increased incidence of lymph node metastasis of HNCs., (2008 S. Karger AG, Basel)

This animal experiment was aimed to apply fibrin in facial nerve repair and to quest for technical improvements in facial surgery. In each of 15 healthy large ear white rabbits, a unilateral 5 mm intratemporal facial nerve gap was created, the proximal and distal stumps were inserted into chitin tube, 1 ml autologous fibrin glue was applied around the anastomotic zone, and no suture was employed. At 3 months and 5 months after opertion, electrophysioligical study was performed. Compared with normal nerves, the regenerating nerves in both the chitin tube bridged group and the perineurium suture group had longer incubation period, lower amplitude, slower nerve-muscle conduction velocity at 3 months postoperatively. The differences were distinctly significant (P < 0.01). Although being decreased at 5 months after operation, the differences were still statistically significant (P < 0.05). There were no significant differences between the chitin tube bridged group and perineurium suture group at 3 months and 5 months, respectively. The study suggests that facial nerve repair using fibrin glue and chitin tube has the advantages of being easier,faster and more stable.

Objective: To observe the changes of GABA and GABAergic neurons in rat inferior colliculus following unilateral cochlear damage and explore the function and significance of GABA in reorganization of auditory center after deafferentation., Method: Twenty sprague dawley rats were divided into four groups randomly. The technique of direct anti-GABA immunocytochemistry (SP) was used in this study. The quantity of GABA was measured by 835-50 type Amino Acid Automatism Analyzer and compared at one week, two weeks and one month after unilateral cochlear ablation, respectively., Result: Compared with controlled groups, the quantity levels of GABA decreased and the number of GABAergic neurons also reduced one week after unilateral cochlear damage and there was a significant difference in GABA between two groups( P <0. 05); after one month, the quantity of GABA and the number of GABAergic neurons almost reached the normal level, the differences were no significant., Conclusion: The dynamic change of GABA in rat inferior colliculus reflected the neuronal activity, which implied that GABA may play an important role in reorganization of auditory center after unilateral cochlear damage.

Objective: To observe the distribution of gama-aminobutyric acid (GABA) ergic neurons in rats cochlear nuclei (CN) in pre-and post-operation of unilateral cochlea ablation., Method: The technique of direct anti-GABA immunocytochemistry (SP) was used in this study., Result: The GABAergic neurons existed extensively in cochlear nuclei(CN). The number was more, the volume was larger of GABAergic cells in CN and those cells were stained heavily before ablation. The number of GABAergic cells in lesion side was far less than that in intact side after unilateral cochlea ablation one to two weeks (P < 0.01), then the number of GABAergic cells increased after three weeks which was still less (P > 0.05) and almost reached normal after a month (P > 0.05)., Conclusion: The changes of GABAergic neurons reflect the balance between excitatory and inhibitory of neurons. It suggests that changes in the extent of GABAergic inhibition have been shown to contribute to the reorganization of primary auditory central system after unilateral cochlea ablation in rats. The decrease and increase of the number of GABAergic-neurons are essential for this reorganization.