Synergistic anti-tumor effects of lenalidomide and gefitinib by upregulating ADRB2 and inactivating the mTOR/PI3K/AKT signaling pathway in lung adenocarcinoma.

Gefitinib is commonly used to be the first-line therapy for advanced non-small cell lung cancer (NSCLC). Therapeutic effect of gefitinib is reduced due to acquired resistance, and combined treatment is recommended. In this research, we planned to explore the impacts of combined treatment of lenalidomide and gefitinib on gefitinib-sensitive or -resistant NSCLC cells. The co-treatment results demonstrated that enhanced antitumor impact on NSCLC cell growth, migration, invasion, cell cycle process and apoptosis. The tumor-bearing mouse models were established using PC9/GR cells. In vivo assays also showed that lenalidomide and gefitinib synergistically inhibited mouse tumor growth along increased the survival of mice. ADRB2 was identified as a lowly expressed gene in PC9/GR cells and LUAD tumor tissues. LUAD patients with high ADRB2 expression were indicated with favorable survival outcomes. Moreover, ADRB2 was upregulated in lenalidomide and/or gefitinib-treated PC9/GR cells. ADRB2 deficiency partially offsets the suppressive impacts of lenalidomide and gefitinib co-treatment on the viability and proliferation of PC9/GR cells. Additionally, lenalidomide and gefitinib cotreatment significantly inactivated the mTOR/PI3K/AKT signaling pathway compared with each treatment alone. Rescue assays were performed to explore whether lenalidomide and gefitinib synergistically inhibited the growth of PC9/GR cells via the PI3K/AKT pathway. PI3K activator SC79 significantly restored reduced cell proliferation, migration and invasion along with elevated cell cycle arrest and apoptosis caused by lenalidomide and gefitinib cotreatment. In conclusion, lenalidomide and gefitinib synergistically suppressed LUAD progression and attenuated gefitinib resistance by upregulating ADRB2 and inactivating the mTOR/PI3K/AKT signaling pathway in lung adenocarcinoma.

Pituitary transcriptome profile from laying period to incubation period of Changshun green-shell laying hens.

BACKGROUND: Incubation behaviour, an instinct for natural breeding in poultry, is strictly controlled by the central nervous system and multiple neuroendocrine hormones and neurotransmitters, and is closely associated with the cessation of egg laying. Therefore, it is essential for the commercial poultry industry to clarify the molecular regulation mechanism of incubation behaviour. Here, we used high-throughput sequencing technology to examine the pituitary transcriptome of Changshun green-shell laying hen, a local breed from Guizhou province, China, with strong broodiness, in two reproductive stages, including egg-laying phase (LP) and incubation phase (BP). We also analyze the differences in gene expression during the transition from egg-laying to incubation, and identify critical pathways and candidate genes involved in controlling the incubation behaviour in the pituitary. RESULTS: In this study, we demonstrated that a total of 2089 differently expressed genes (DEGs) were identified in the pituitary, including 842 up-regulated and 1247 down-regulated genes. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed that steroid biosynthesis pathway and neuroactive ligand-receptor interaction were significantly enriched based on DEGs commonly identified in pituitary. Further analysis revealed that SRC, ITGB4, ITGB3, PIK3R3 and DRD2 may play crucial roles in the regulation of incubation behaviour. CONCLUSIONS: We identified 2089 DEGs and the key signaling pathways which may be closely correlated with incubation in Changshun green-shell laying hens, and clarified the molecular regulation mechanism of incubation behaviour. Our results indicate the complexity and variety of differences in reproductive behaviour of different chicken breeds.

Immune-related thyroid dysfunction in patients with non-small cell lung cancer.

Background: Immune-related thyroid dysfunction (irTD) is a common immune-related adverse event (irAE). The potential biomarkers of irTDs and their impact on the clinical outcomes of patients with non-small cell lung cancer (NSCLC) treated with immune checkpoint inhibitors (ICIs) remain unclear. We aimed to identify potential biomarkers of irTDs and reveal the association between irTDs and the clinical outcomes in patients with NSCLC treated with ICIs. Methods: We conducted a retrospective study on 126 patients with NSCLC, who were treated with pembrolizumab, sintilimab, atezolizumab, or camrelizumab, as first-line therapy, at the First Affiliated Hospital, College of Medicine, Zhejiang University, between July 2019 and February 2023. Anti-thyroid antibodies (ATAs), thyroid peroxidase antibody (TPOAb), thyroglobulin antibody (TGAb), serum interleukin-6 (IL-6), thyroid ultrasonography, overall survival (OS), and progression-free survival (PFS) were the main indicators. Results: Most (92.9%) irTD cases occurred no later than one year after ICIs initiation. Patients with irTDs had higher positive rates for ATAs and TPOAb [33.3% vs. 1.3%, and 30.3% vs. 1.3%, both P<0.01, odds ratio (OR) =39.81, and OR =35.46, respectively]. Irregular echo pattern and diffuse changes were more common in patients with irTDs (70.7% vs. 47.2%, and 19.5% vs. 1.4%, P<0.05 and P<0.01, OR =2.70, and OR =17.21, respectively). OS and PFS were similar in patients with and without irTDs (P>0.05). Conclusions: The ATAs, TPOAb, and abnormal thyroid ultrasonographic findings (irregular echo patterns and diffuse changes) are potential biomarkers of irTDs. Patients with NSCLC treated with ICIs (pembrolizumab, sintilimab, atezolizumab, and camrelizumab) who developed irTDs had no advantage in terms of clinical outcomes compared to euthyroid patients.

The response of rhubarb to smut infection is revealed through a comparative transcriptome and metabolome study.

MAIN CONCLUSION: Integrated transcriptome and metabolome analysis have unveiled the physiological and molecular responses of rhubarb to infection by smut fungi. Rhubarb is an important medicinal plant that is easily infected by smut fungi during its growth. Thus far, no research on the influence of smut fungi on the growth of rhubarb and its secondary metabolism has been conducted. In this study, petioles of Chinese rhubarb (Rheum officinale) [healthy or infected with smut fungus (Thecaphora schwarzmaniana)] were characterized. Microscopic structure, global gene expression profiling, global metabolic profiling, and key enzyme activity and metabolite levels in infected plants were analyzed. Infection by smut fungi resulted in numerous holes inside the petiole tissue and led to visible tumors on the external surface of the petiole. Through metabolic changes, T. schwarzmaniana induced the production of specific sugars, lipids, and amino acids, and inhibited the metabolism of phenolics and flavonoids in R. officinale. The concentrations of key medicinal compounds (anthraquinones) were decreased because of smut fungus infection. In terms of gene expression, the presence of T. schwarzmaniana led to upregulation of the genes associated with nutrient (sugar, amino acid, etc.) transport and metabolism. The gene expression profiling showed a stimulated cell division activity (the basis of tumor formation). Although plant antioxidative response was enhanced, the plant defense response against pathogen was suppressed by T. schwarzmaniana, as indicated by the expression profiling of genes involved in biotic and abiotic stress-related hormone signaling and the synthesis of plant disease resistance proteins. This study demonstrated physiological and molecular changes in R. officinale under T. schwarzmaniana infection, reflecting the survival tactics employed by smut fungus for parasitizing rhubarb.

Detection of Bartonella in kissing bugs Triatoma rubrofasciata collected from Huizhou City, South China.

Background: The blood-feeding behavior of kissing bugs (subfamily Triatominae, family Reduviidae, order Hemiptera) means they are potential vectors of multiple humans pathogens. However, investigations of vector-borne pathogens harbored by kissing bugs are rare. Methods: In the current study, 22 adult kissing bugs (Triatoma rubrofasciata) were captured in Huizhou City, Guangdong Province, south China. The presence of vector-borne pathogens in the kissing bugs was tested, and the genetic diversity of these potential pathogens was investigated. Results: All the kissing bugs were negative for Anaplasmataceae bacteria, Rickettsia, and Coxiella. Bartonella DNA was detected in 36.4% (8/22) of the kissing bugs. The sequences of the Bartonella gltA genes divided into two clades in a phylogenetic tree, with close relationships to B. tribocorum and uncultured Bartonella sp. clone MYR-283, respectively. All the groEL sequences were closely related to those of B. kosoyi (identity 98.75%-100%). The ftsZ and rpoB sequences were most closely related to those of B. elizabethae, a recognized human pathogen, with nucleotide similarities of 98.70%-100% and 99.45%-100%, respectively. Conclusions: We report the detection of Bartonella DNA in Triatoma kissing bugs in southern China. Although the sample size is limited, the high positive rate of detection of Bartonella DNA, the close relationship of the gene sequences to those of zoonotic Bartonella species, and the distribution of the kissing bugs near human residences, hint at a risk to public health.

The role of TMEM26 in disrupting tight junctions and activating NF-κB signaling to promote epithelial-mesenchymal transition in esophageal squamous cell carcinoma.

OBJECTIVES: Metastasis is one of the biggest challenges in the management of Esophageal Squamous Cell Carcinoma (ESCC), of which molecular mechanisms remain elusive. The present study aimed to explore the roles and underlying mechanisms of Transmembrane protein 26 (TMEM26) in ESCC. METHOD: TMEM26 expressions in tumorous and adjacent tissues from patients with ESCC and in normal esophageal epithelial and ESCC cell lines were detected by immunostaining and western blotting, respectively. The Epithelial-Mesenchymal Transition (EMT), a critical process during metastasis, was investigated by wound healing and Transwell assays, and EMT-related proteins were examined after the TMEM26 alteration in ESCC cell lines. NF-κB signaling activation and Tight Junction (TJ) protein expression were analyzed by western blotting and immunofluorescence, respectively. In vivo verification was performed on the liver metastatic murine model. RESULTS: Compared with non-cancerous esophageal tissues and cells, the TMEM26 expression level was higher in ESCC samples and cell lines, where the plasma membrane localization of TMEM26 was observed. The EMT-related processes of ESCC cells were suppressed by RNAi depletion of TMEM26 but aggravated by TMEM26 overexpression. Mechanistically, TMEM26 promoted NF-κB signaling to accelerate EMT in ESCC cells. The plasma membrane presentation and assembly of TJ proteins were impaired by TMEM26. CONCLUSION: Overall, TMEM26 acts as a critical determinant for EMT in ESCC cells by disrupting TJ formation and promoting NF-κB signaling, which may be a potential therapeutic target for treating metastatic ESCC.

Polysaccharides from Bletilla striata protect against mercury-induced gastrointestinal toxicology in adult Drosophila melanogaster via modulation of sestrin.

Oxidative stress was one of the major causes of heavy metal-induced toxicity in organisms. The polysaccharide from Bletilla striata (Orchidaceae) (BSP) has been recently recognized as a novel player in the management of oxidative stress response in organisms. Here, we took the midgut of adult Drosophila melanogaster (Diptera: Drosophilidae) (D. melanogaster), a functional equivalent to the mammalian intestine and stomach, as a model to evaluate the protective effects of BSP (50 µg/mL) on mercuric chloride-induced gastrointestinal toxicology in insects. As a result, BSP exposure significantly improved the survival rates and climbing ability of adult flies exposed to mercury. Further study demonstrated that BSP significantly alleviated the mercury-induced oxidative injury to midgut epithelium, at least partly, through increasing antioxidant enzyme activity (glutathione-S-transferase and superoxide dismutase), decreasing reactive oxidative species production, inhibiting cell death, restoring intestinal epithelial barrier and regulating intestinal stem cell-mediated tissue regeneration. Additionally, sestrin, an oxidative-stress gene, was required in mediating the protection of BSP against mercury-induced oxidative damage to midgut. This study suggested that BSP has great potential for future application in the treatment and prevention of heavy metal-induced gastrointestinal adversities in mammals.

Demulsification Performance and Mechanism of Tertiary Amine Polymer-Grafted Magnetic Nanoparticles in Surfactant-Free Oil-in-Water Emulsion.

Numerous cationic magnetic nanoparticles (MNPs) have previously been developed for demulsifying oil-in-water (O/W) emulsion, and results showed that the cationic MNPs could effectively flocculate and remove the negatively charged oil droplets via charge attraction; however, to the best of our knowledge, there are no research reports regarding the synergetic influence of both the positive charge density and interfacial activity of MNPs on the demulsification performance. In this study, three tertiary amine polymer-grafted MNPs, namely, poly(2-dimethylaminoethyl acrylate)-grafted MNPs (M-PDMAEA), poly(2-dimethylamino)ethyl methacrylate)-grafted MNPs (M-PDMAEMA), and poly(2-diethylaminoethyl methacrylate)-grafted MNPs (M-PDEAEMA), were synthesized and evaluated for their demulsification performance. Results demonstrated that a high positive charge density and superior interfacial activity of MNPs could cause partial oil droplet re-dispersion when excessive MNPs were introduced, leading to a lower magnetic separation efficiency and narrower demulsification window. Herein, a demulsification window is defined as a range of nanoparticle dosages in which the MNPs can effectively demulsify the O/W emulsion under certain conditions. For highly positively charged MNPs, their good interfacial activity could aggravate the formation of a narrower demulsification window. When tertiary amine polymer-grafted MNPs carried a lower positive charge density or weak interfacial activity, that is, M-PDMAEA at pH 4.0, M-PDMAEMA at pH 5.0-9.0, and M-PDEAEMA at pH 9.0-10.0, wide demulsification windows were observed. Additionally, a recycling experiment suggested that MNPs could maintain high demulsification efficiency up to at least five cycles, indicating their satisfactory recyclability. The three tertiary amine polymer-grafted MNPs can be potentially used for efficient demulsification from surfactant-free O/W emulsion in various pH ranges.

Chronic exposure to zearalenone induces intestinal inflammation and oxidative injury in adult Drosophila melanogaster midgut.

In the past decade, mycotoxin zearalenone (ZEN)-induced gastrointestinal adverse effects have been increasingly attracting worldwide attention. This study aimed to determine the gastrointestinal adverse effects of ZEN in Drosophila melanogaster (D. melanogaster) and reveal possible mechanisms of action of ZEN in insects. Here, chronic exposure of D. melanogaster to ZEN killed flies in a dose-dependent manner (2-20 µM). ZEN (20 µM) decreased the survival rates and climbing ability of flies, and activated immune deficiency-mediated intestinal immunity in midgut, leading to the production of antimicrobial peptides. Meanwhile, ZEN exposure induced morphological alteration of adult midgut. Further study suggested that high levels of oxidative stress was observed in ZEN-treated midgut due to the imbalance between the production of reactive oxygen species and the expression and activities of cellular antioxidant enzyme, including superoxide dismutase and catalase. ZEN-induced oxidative stress then caused cell death, impaired gut barrier function and increased gut permeability, leading to oxidative injury in midgut. Subsequently, ZEN-induce midgut injury further disrupted intestinal stem cell (ISC) homeostasis, stimulating ISC proliferation and tissue regeneration, but did not alter cell fate specification of ISC. Additionally, activation of Jun N-terminal kinase pathway was involved in ZEN-induced oxidative injury and tissue regeneration in midgut. Antioxidant vitamin E alleviated ZEN-induced oxidative injury to midgut epithelium. Collectively, this study provided additional evidences for ZEN-induced gastrointestinal adverse effects from an invertebrate model, extended our understanding of the mechanisms mediating mycotoxin toxicity in organisms.

The role of TMEM26 in disrupting tight junctions and activating NF-κB signaling to promote epithelial-mesenchymal transition in esophageal squamous cell carcinoma

Abstract Objectives Metastasis is one of the biggest challenges in the management of Esophageal Squamous Cell Carcinoma (ESCC), of which molecular mechanisms remain elusive. The present study aimed to explore the roles and underlying mechanisms of Transmembrane protein 26 (TMEM26) in ESCC. Method TMEM26 expressions in tumorous and adjacent tissues from patients with ESCC and in normal esophageal epithelial and ESCC cell lines were detected by immunostaining and western blotting, respectively. The Epithelial-Mesenchymal Transition (EMT), a critical process during metastasis, was investigated by wound healing and Transwell assays, and EMT-related proteins were examined after the TMEM26 alteration in ESCC cell lines. NF-κB signaling activation and Tight Junction (TJ) protein expression were analyzed by western blotting and immunofluorescence, respectively. In vivo verification was performed on the liver metastatic murine model. Results Compared with non-cancerous esophageal tissues and cells, the TMEM26 expression level was higher in ESCC samples and cell lines, where the plasma membrane localization of TMEM26 was observed. The EMT-related processes of ESCC cells were suppressed by RNAi depletion of TMEM26 but aggravated by TMEM26 overexpression. Mechanistically, TMEM26 promoted NF-κB signaling to accelerate EMT in ESCC cells. The plasma membrane presentation and assembly of TJ proteins were impaired by TMEM26. Conclusion Overall, TMEM26 acts as a critical determinant for EMT in ESCC cells by disrupting TJ formation and promoting NF-κB signaling, which may be a potential therapeutic target for treating metastatic ESCC.

A stable and sensitive Au metal organic frameworks resonance Rayleigh scattering nanoprobe for detection of SO3 2- in food based on fuchsin addition reaction.

A stable Au metal organic frameworks (AuMOF) nanosol was prepared. It was characterized by electron microscopy and molecular spectral techniques. In pH 6.8 PBS buffer solution, AuMOF nanoprobes exhibit a strong resonance Rayleigh scattering (RRS) peak at 330 nm. After basic fuchsin (BF) adsorbing on the surface of AuMOF, the RRS energy of the nanoprobe donor can be transferred to BF receptor, resulting in a decrease in the RRS intensity at 330 nm. Both sulfite and BF taken place an addition reaction to form a colorless product (SBF) that exhibit weak RRS energy transfer (RRS-ET) between AuMOF and SBF, resulting in the enhancement of the RRS peak. As the concentration of SO3 2-increases, the RRS peak is linearly enhanced. Thus, a new and sensitive RRS-ET method for the detection of SO3 2- (0.160-5.00 µmol/L) was developed accordingly using AuMOF as nanoprobes, with a detection limit of 0.0800 µmol/L. This new RRS method was applied to determination of SO3 2- in food and SO2 in air samples. The recoveries of food and air samples were 97.1-106% and 92.9-106%, and the relative standard deviation (RSD) was 2.10-4.80% and 2.10-4.50%, respectively.

Synergy evaluation of China's economy-energy low-carbon transition and its improvement strategy for structure optimization.

Low-carbon economic development and energy transition are interactively linked. The synergetic development of the two subsystems is important to achieve the "double carbon" goal of sustainable development. First, this study proposes a model to measure the current synergy level of China's economy-energy low-carbon transition. Second, an optimization model is developed to improve industry and energy synergy levels through structure optimization. The synergy degree (SD) level of China's economy-energy low-carbon transition increased from 0 to 0.98 between 2005 and 2017. Furthermore, 69.2% of the periods are in a state of asynergy (SD < 0.6). By implementing the industry and energy structure optimization (OPT) scenario, the synergy level by 2035 can be 27.8% higher than the business-as-usual (BAU) scenario. Moreover, light synergy (0.6 ≤ SD < 0.8) could be achieved by 2025, and high-quality synergy (0.9 ≤ SD ≤ 1) by 2033 in the OPT scenario. Conversely, the synergy level can only achieve light synergy until 2035 in the BAU scenario. Compared to energy structure optimization, the low carbonization of the economic structure plays a more significant role in improving the synergy level of the transaction. These findings can provide support for China's policy-making regarding economic and energy transition.

Numerical Analysis of Thermal Radiative Maxwell Nanofluid Flow Over-Stretching Porous Rotating Disk.

The fluid flow over a rotating disk is critically important due to its application in a broad spectrum of industries and engineering and scientific fields. In this article, the traditional swirling flow of Von Karman is optimized for Maxwell fluid over a porous spinning disc with a consistent suction/injection effect. Buongiorno's model, which incorporates the effect of both thermophoresis and Brownian motion, describes the Maxwell nanofluid nature. The dimensionless system of ordinary differential equations (ODEs) has been diminished from the system of modeled equations through a proper transformation framework. Which is numerically computed with the bvp4c method and for validity purposes, the results are compared with the RK4 technique. The effect of mathematical abstractions on velocity, energy, concentration, and magnetic power is sketched and debated. It is perceived that the mass transmission significantly rises with the thermophoresis parameter, while the velocities in angular and radial directions are reducing with enlarging of the viscosity parameter. Further, the influences of thermal radiation Rd and Brownian motion parameters are particularly more valuable to enhance fluid temperature. The fluid velocity is reduced by the action of suction effects. The suction effect grips the fluid particles towards the pores of the disk, which causes the momentum boundary layer reduction.

HHIP Overexpression Suppresses Human Gastric Cancer Progression and Metastasis by Reducing Its CpG Island Methylation.

Human hedgehog-interacting protein (HHIP), a negative regulator of hedgehog (HH) signaling pathway, has been reported to be dysregulated in many types of cancer, including gastric cancer. However, the inhibitory role of HHIP as well as the underlying molecular mechanism of HHIP regulation in gastric cancer haven't been fully elucidated yet. In this study, we demonstrated that HHIP overexpression significantly suppressed the proliferation and invasion of AGS cells evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and transwell assays, respectively. Interestingly, methylation-specific polymerase chain reaction (MS-PCR, MSP) showed that HHIP overexpression dramatically decreased its de novo promoter methylation levels in AGS cells. Furthermore, HHIP expression was higher in adjacent non-cancerous tissue compared to matched gastric cancer tissue. High HHIP level was negatively correlated with metastasis (p = 0.035) but not local recurrence (p = 0.58). Taken together, our study suggested that HHIP can modulate gastric cancer progression and metastasis via regulation of its de novo promoter methylation levels in a feedback manner. Lower HHIP levels is positively associated with gastric cancer metastasis, which not only indicates HHIP could be served as a protective marker for gastric cancer, but also suggests restoring HHIP expression might be a potential therapeutic strategy for clinical treatment.

RETINAL MICROVASCULATURE ALTERATION IN ACTIVE THYROID-ASSOCIATED OPHTHALMOPATHY.

OBJECTIVE: It is uncertain whether there is an increased retinal vessel density in patients with thyroid-associated ophthalmology (TAO), or if ocular hemodynamic changes are associated with their vision. Many techniques have been applied to analyze ocular hemodynamic changes in TAO patients; however, they suffer from limitations. Optical coherence tomography angiography (OCTA) is more sensitive to hemodynamic changes. We performed OCTA to determine whether there is an increased retinal vessel density in TAO patients, and investigate associations with their vision. METHODS: Twenty TAO patients (right eye) and 20 normal controls (right eye) were recruited for the study. Three-dimensional OCTA images were generated with 6 × 6-mm angiographic OCT scans of the superficial retinal layer (SRL) and deeper retinal layer (DRL) of the macular retina for each eye. We calculated the microvascular (MIR) and macrovascular (MAR) densities and compared MIR, MAR, and total MIR (TMI) density in the healthy control and TAO groups separately. We used the annular partition (C1-C6) and quadrant partition method to analyze retinal vessel density alterations. Correlation analyses were used to examine the association of MIR density and visual acuity. RESULTS: In the SRL, STMI (superficial TMI) and SMIR (superficial MIR) density increased in the macular area of TAO patients ( P<.05). For the DRL, the density of DMIR (deeper MIR) increased in macular area only ( P<.05). However, with the annular partition method, TAO patients showed significantly increased SMIR density in the C1 and C2 regions ( P<.05), and increased DMIR density only in the C1 region ( P<.05). In the SRL, the correlation coefficient of the STMI/SMIR density in the TAO group was -0.538 ( r = -0.538, P = .032), and visual acuity was -0.624 ( r = -0.624, P = .010). CONCLUSION: OCTA findings suggest that active TAO patients present with an increased retinal MIR density. Moreover, increased macular STMI and SMIR density might affect visual acuity. ABBREVIATIONS: CDI = color Doppler imaging; DMIR = deeper MIR; DRL = deeper retinal layer; EDV = end diastolic velocity; FAZ = foveal avascular zone; GO = Graves ophthalmopathy; MAR = macrovascular; MIR = microvascular; OCT = optical coherence tomography; OCTA = optical coherence tomography angiography; RI = resistance index; ROC = receiver operating characteristic; SMIR = superficial MIR; SOV = superior ophthalmic vein; SRL = superficial retinal layer; SSADA = split-spectrum amplitude-decor-relation angiography; STMI = superficial TMI; TAO = thyroid-associated ophthalmology; TMI = total MIR.

Bradykinin potentially stimulates cell proliferation in rabbit corneal endothelial cells through the ZO­1/ZONAB pathway.

Bradykinin (BK) has been demonstrated to induce proliferation in several types of cell in ex vivo corneas. However, the mechanisms underlying the action of BK on corneal endothelial cells (CECs) remain largely unknown. The present study aimed to investigate the effect of BK on rabbit corneal endothelial cell (RCEC) proliferation, and assess the involvement of the zonula occludens­1(ZO­1)/ZO­1associated nucleic acid binding protein (ZONAB) pathway. Cell proliferation and cell cycle distribution was analyzed following treatment with BK (0.01, 0.1,1.0 or 10.0 µM) for the indicated time intervals (24, 48, 72 and 96 h), or following BK treatment combined with transfection of ZONAB­small interfering (si)RNA for 72 h. In addition, the expression of tight junction ZO­1, nuclear ZONAB, proliferating cell nuclear antigen(PCNA) and cyclin D1 were evaluated using western blotting or immunofluorescence. BK treatment was demonstrated to induce time­ and concentration­dependent cell proliferation and cell cycle progression, along with the upregulation of tight junction ZO­1 and nuclear ZONAB, as well as PCNA and cyclin D1 protein expression. Furthermore, knockdown with ZONAB­siRNA inhibited cell proliferation, induced cell cycle arrest and downregulated PCNA and cyclin D1 protein expression. ZONAB knockdown therefore successfully reversed the increase in proliferation induced by BK treatment. Taken together, these results suggested that BK stimulated RCEC proliferation, potentially via the ZO­1/ZONAB pathway. The signaling paradigm disclosed in the present study potentially serves as an important therapeutic target for cornea regeneration and transplantation.

Ecological Effect of Arginine on Oral Microbiota.

Dental caries is closely associated with the microbial dybiosis between acidogenic/aciduric pathogens and alkali-generating commensal bacteria colonized in the oral cavity. Our recent studies have shown that arginine may represent a promising anti-caries agent by modulating microbial composition in an in vitro consortium. However, the effect of arginine on the oral microbiota has yet to be comprehensively delineated in either clinical cohort or in vitro biofilm models that better represent the microbial diversity of oral cavity. Here, by employing a clinical cohort and a saliva-derived biofilm model, we demonstrated that arginine treatment could favorably modulate the oral microbiota of caries-active individuals. Specifically, treatment with arginine-containing dentifrice normalized the oral microbiota of caries-active individuals similar to that of caries-free controls in terms of microbial structure, abundance of typical species, enzymatic activities of glycolysis and alkali-generation related enzymes and their corresponding transcripts. Moreover, we found that combinatory use of arginine with fluoride could better enrich alkali-generating Streptococcus sanguinis and suppress acidogenic/aciduric Streptococcus mutans, and thus significantly retard the demineralizing capability of saliva-derived oral biofilm. Hence, we propose that fluoride and arginine have a potential synergistic effect in maintaining an eco-friendly oral microbial equilibrium in favor of better caries management.

Analysis of chemosensory gene families in the beetle Monochamus alternatus and its parasitoid Dastarcus helophoroides.

We assembled antennal transcriptomes of pest Monochamus alternatus and its parasitoid Dastarcus helophoroides to identify the members of the major chemosensory multi-gene families. Gene ontology (GO) annotation indicated that the relative abundance of transcripts associated with specific GO terms was highly similar in the two species. In chemosensory gene families, we identified 52 transcripts encoding putative odorant-binding proteins (OBPs), 19 chemosensory proteins (CSPs), 10 olfactory receptors (ORs), 8 ionotropic receptors (IRs), 2 gustatory receptors (GRs), and 5 sensory neuron membrane proteins (SNMPs) in these two transcriptomes. Predicted protein sequences were compared with Dendroctonus ponderosae, Tribolium castaneum and Drosophila melanogaster. The results of phylogenetic trees showed that some clusters included only OBPs or CSPs from D. helophoroides, some clusters included only OBPs or CSPs from M. alternatus, while some clusters included OBPs or CSPs from both M. alternatus and D. helophoroides. The identification of the chemosensory genes and the phylogenetic relationship of these genes between two species might provide new ideas for controlling M. alternatus and improving current strategies for biological control.

De novo analysis of the Nilaparvata lugens (Stål) antenna transcriptome and expression patterns of olfactory genes.

We sequenced the antenna transcriptome of the brown planthopper (BPH), Nilaparvata lugens (Stål), a global rice pest, and performed transcriptome analysis on BPH antenna. We obtained about 40million 90bp reads that were assembled into 75,874 unigenes with a mean size of 456bp. Among the antenna transcripts, 32,856 (43%) showed significant similarity (E-value <1e(-5)) to known proteins in the NCBI database. Gene ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were used to classify functions of BPH antenna genes. We identified 10 odorant-binding proteins (OBPs), including 7 previously unidentified, and 11 chemosensory proteins (CSPs), including two new members. The expression profiles of 4 OBPs and 2 CSPs were determined by q-PCR for antenna, abdomen, leg and wing of insects of different age, gender, and mating status including two BPH adult wing-morphology types. NlugCSP10 and 4 OBPs appeared to be antenna-specific because they were highly and differentially expressed in male and female antennae. NlugCSP11 was expressed ubiquitously, with particularly high expression in wings. The transcript levels of several olfactory genes depended on adult wing form, age, gender, and mating status, although no clear expression patterns were determined.

Transgenic Bt rice does not challenge host preference of the target pest of rice leaffolder, Cnaphalocrocis medinalis (Lepidoptera: Pyralidae).

BACKGROUND: Transgenic Bt rice line T2A-1 expresses a synthesized cry2A gene that shows high resistance to Lepidoptera pests, including Cnaphalocrocis medinalis (Guenée) (Lepidoptera: Pyralidae). Plant volatile orientation cues and the physical characteristics of the leaf surface play key roles in host location or host-plant acceptance of phytophagous insects. These volatile compounds and physical traits may become altered in Bt rice and it is not known whether this influences the behavior of C. medinalis when searching for oviposition sites. RESULTS: The results of electronic nose analysis showed that the Radar map of Bt rice cultivars was analogous to the non- Bt rice cultivars at each growing stage. PCA analysis was able to partly discriminate between some of the Bt vs. non-Bt rice sensors, but could not to separate Bt cultivars from non-Bt cultivars. The total ion chromatogram between Bt and non-Bt rice cultivars at the seedling, booting and tillering stages were similar and 25 main compounds were identified by GC-MS. For most compounds, there was no significant difference in compound quantities between Bt and non-Bt rice cultivars at equivalent growth stages. The densities of the tubercle papicles and the trichomes on the upper and lower surfaces were statistically equal in Bt and non-Bt rice. The target pest, C. medinalis, was attracted to host rice plants, but it could not distinguish between the transgenic and the isogenic rice lines. CONCLUSIONS: There were no significant differences between the Bt rice line, T2A-1 and the non-Bt rice for volatiles produced or in its physical characteristics and there were no negative impacts on C. medinalis oviposition behavior. These results add to the mounting evidence that Bt rice has no negative impact on the target insect oviposition behavior.