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Background: Previous studies have demonstrated that long non-coding RNA maternally expressed gene 3 (MEG3) emerged as a key regulator in development and tumorigenesis. This study aims to investigate the function and mechanism of MEG3 in osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) and explores the use of MEG3 in skull defects bone repairing., Methods: Endogenous expression of MEG3 during BMSCs osteogenic differentiation was detected by quantitative real-time polymerase chain reaction (qPCR). MEG3 was knockdown in BMSCs by lentiviral transduction. The proliferation, osteogenic-related genes and proteins expression of MEG3 knockdown BMSCs were assessed by Cell Counting Kit-8 (CCK-8) assay, qPCR, alizarin red and alkaline phosphatase staining. Western blot was used to detect β-catenin expression in MEG3 knockdown BMSCs. Dickkopf 1 (DKK1) was used to block wnt/β-catenin pathway. The osteogenic-related genes and proteins expression of MEG3 knockdown BMSCs after wnt/β-catenin inhibition were assessed by qPCR, alizarin red and alkaline phosphatase staining. MEG3 knockdown BMSCs scaffold with PHMG were implanted in a critical-sized skull defects of rat model. Micro-computed tomography(micro-CT), hematoxylin and eosin staining and immunohistochemistry were performed to evaluate the bone repairing., Results: Endogenous expression of MEG3 was increased during osteogenic differentiation of BMSCs. Downregulation of MEG3 could promote osteogenic differentiation of BMSCs in vitro. Notably, a further mechanism study revealed that MEG3 knockdown could activate Wnt/β-catenin signaling pathway in BMSCs. Wnt/β-catenin inhibition would impair MEG3-induced osteogenic differentiation of BMSCs. By using poly (3-hydroxybutyrate-co-3-hydroxyhexanoate, PHBHHx)-mesoporous bioactive glass (PHMG) scaffold with MEG3 knockdown BMSCs, we found that downregulation of MEG3 in BMSCs could accelerate bone repairing in a critical-sized skull defects rat model., Conclusions: Our study reveals the important role of MEG3 during osteogenic differentiation and bone regeneration. Thus, MEG3 engineered BMSCs may be effective potential therapeutic targets for skull defects.

Bluetongue is an arthropod-borne viral disease of ruminants caused by bluetongue virus (BTV). In China, BTV is relatively common in Yunnan Province with the exception of northern regions around Shangri-La, where the average altitude is approximately 3,450 metres. Recently, the seroprevalence of BTV has been measured in yaks in Shangri-La; therefore, this study investigated BTV infections in this area. The serological investigation in five villages in Shangri-La showed that there were sporadic BTV infections in yaks (20 of 507 positive) during 2014 to 2017, while the seroprevalence of BTV at three goat farms in a nearby river valley was 35%-65% in 2017. Subsequently, 20 sentinel goats were kept on two separate farms in the river valley and monitored for seroconversion between May and September of 2017. Five of the sentinel animals were tested positive for antibodies to BTV by C-ELISA during the study period, and 13 BTV isolates were isolated from ten sentinel animals. All isolates were identified as the same serotype, and the complete nucleotide sequence of one was determined. The genomic sequences showed that the isolated BTV strain belonged to serotype 21 and had approximately 99.8%-100% homology with three Indonesian BTV-21 strains (D151, RIVS-66 and RIVS-60) between their coding sequences (CDSs) except for Seg4 (99.5%). Besides, our data suggested that this BTV-21 strain might have also infected some local yaks and sheep., (© 2019 The Authors. Transboundary and Emerging Diseases Published by Blackwell Verlag GmbH.)

Background: Bluetongue disease of ruminants is a typical insect-borne disease caused by bluetongue virus (BTV) of the genus Orbivirus (family Reoviridae) and transmitted by some species of Culicoides (Diptera: Ceratopogonidae). Recently, the detection of BTV in yaks in high altitude meadows of the Shangri-La district of Yunnan Province, China, prompted an investigation of the Culicoides fauna as potential vectors of BTV., Methods: A total of 806 Culicoides midges were collected by light trapping at three sites at altitudes ranging from 1800 to 3300 m. The species were identified based on morphology and the DNA sequences of cytochrome c oxidase subunit 1 (cox1). PCR and quantitative PCR following reverse transcription were used to test for the presence of BTV RNA in Culicoides spp. A phylogenetic analysis was used to analyze the cox1 sequences of some specimens., Results: Four species dominated these collections and cox1 barcoding revealed that at least two of these appear to belong to species new to science. Culicoides tainanus and a cryptic species morphologically similar to C. tainanus dominated low altitude valley collections while C. nielamensis was the most abundant species in the high-altitude meadow. A species related to C. obsoletus occurred at all altitudes but did not dominate any of the collections. BTV RT-qPCR analysis detected BTV RNA in two specimens of C. tainanus, in one specimen closely related to C. tainanus and in one specimen closely related to C. obsoletus by barcode sequencing., Conclusions: This study suggests that BTV in high altitude areas of Yunnan is being transmitted by three species of Culicoides, two of which appear to be new to science. This research may be useful in improving understanding of the effects of global warming on arboviral disease epidemiology and further study is important in research into disease control and prevention.

Background: Difficult airway remains not only a challenge to the anesthesiologists, but also a life-threatening event to the patients. Awake intubation is the principal choice to deal with difficult airway, and a key point for awake intubation is airway topical anesthesia. Yet, so far there is no ideal topical anesthesia approach for awake intubation. This study aimed at evaluating the effect of pressure-driven (by 10 L/min oxygen flow) lidocaine spray on airway topical anesthesia in order to find a powerful and convenient method for airway topical anesthesia for conscious sedation intubation., Methods: Thirty adult patients referred for elective surgery under general anesthesia, aged 18 - C60 years and Mallampati class I or II, were recruited for the study. Before topical anesthesia, the observer's assessment of alert and sedation (OAA/S) scale was controlled between 3 and 4 by intravenous midazolam (0.03 mg/kg), propofol (2 mg×kg(-1)×h(-1)) and remifentanil (0.05 µg×kg(-1)×min(-1)). Ten minutes after sedation, topical anesthesia was performed with the pressure-driven lidocaine spray; the driving pressure was achieved by an oxygen flow of 10 L/min. After topical anesthesia, tracheal intubation was performed and the intubation condition was assessed with modified the Erhan's intubation condition score by an experienced anesthesiologist, and a score of less than 10 was considered to be satisfactory. Attempts to intubate the patient were recorded, and the complications such as local anesthetic toxicity, mucosa injury, and respiration depression were also recorded. The mean arterial blood pressure (MAP), heart rate (HR) and pulse oxygen saturation (SpO2) were recorded at different time points before and after intubation. Patients were asked 24 hours after the operation whether they could recall the events during intubation., Results: All patients were intubated at the first attempt, the average intubation condition score was 7.0 ± 1.1, from 6 to 10, satisfied intubation condition. MAP and HR increased significantly but mildly immediately after the tracheal intubation (P < 0.05), and decreased to the pre-intubation level soon after intubation. There were no related complications and patients had no recall of the intubation procedures., Conclusions: Topical anesthesia with pressure driven 2% lidocaine spray, where pressure is achieved by 10 L/min oxygen flow, can offer satisfactory intubation conditions for conscious sedation intubation.

Aim: To evaluate the antiapoptotic effect of the A20 gene in primary hippocampal neurons both in vivo and in vitro., Methods: Primary hippocampal neurons in embryonic day 18 (E18) rats were transfected with the A20 gene by using the new Nucleofector electroporation transfection method. We then examined, whether A20-neurons possessed anti-apoptotic abilities after TNF-alpha stimulation in vitro. A20-neurons and pcDNA3-neurons were transplanted into the penumbra of the brains of rats that had been subjected to 90-min of ischemia induced by left middle cerebral artery occlusion (MCAO)., Results: A20-neurons resisted TNF-alpha induced apoptosis in vitro. The apoptosis rate of neurons overexpressing A20 (28.46%+/-3.87%) was lower than that in neurons transfected with pcDNA3 (53.06%+/-5.36%). More A20-neurons survived in the penumbra both 3-d and 7-d after transplantation than did sham pcDNA3 neurons., Conclusion: The novel function of A20 may make it a potential targets for the gene therapy for neurological diseases.