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Effect of amantadine dimer adjuvant on humoral immune response induced by SARS-CoV-2 protein vaccine in mice@#Objective To investigate the effect of amantadine dimer adjuvant on humoral immune response induced by SARS-CoV-2 crown protein vaccine in mice.Methods The amantadine dimer was synthesized by substitution reaction ligation,hydrolytic acidification reaction ligation and amide condensation reaction ligation,with which as adjuvant,female BALB/c mice were immunized with the receptor-binding domain(RBD).The mice were randomly divided into five groups,six for each as follows:R6A+RBD group[21 μg(0.033 μmol)amantadine dimer+10 μg RBD],Ada+RBD group[10 μg(0.066 μmol)amantadine+10 μg RBD],Alu+RBD group(35 μg aluminum adjuvant+10 μg RBD),RBD group(10 μg RBD)and Blank group(0.9% normal saline),which were immunized i.m.on day 0,14 and 28 respectively.Serum samples were collected from tail vein of mice 7 d after the second dose and 14 d after the last dose and determined for specific IgG antibody levels by ELISA.Results The amantadine dimer was purified by thin layer chromatography(TLC)and identified by electrospray ionization-MS(ESI-MS)positive/negative ion mode.After two times of immunization,the antibody levels in sera at various dilutions of mice in R6A+RBD group were all higher than those of Ada+RBD group,while lower than those of Alu+RBD group.However,after three times of immunization,the antibody levels in sera at various dilutions of mice in R6A+RBD group were all significantly higher than those of Ada+RBD and Alu+RBD groups(each F > 30,each P < 0.000 1 and each P < 0.01).Conclusion Amantadine dimer adjuvant enhanced humoral immune response induced by SARS-CoV-2 protein vaccine in mice with good adjuvant effect,which may be used as an alternative adjuvant.This strategy based on existing drug transformation provided a new idea for the development of novel adjuvants.
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OBJECTIVE To study the extraction and enrichment technology of chrysosplenides A （CA） and I （CI） in Tibetan medicine Chrysosplenium axillare. METHODS HPLC method was used to determine the contents of CA and CI. The orthogonal experiment was used to optimize the extraction technology of CA and CI in C. axillare using total transfer rate of CA and CI as evaluation indexes， with volume fraction of ethanol， extraction temperature， extraction times and solid-liquid ratio as factors. The validation test was also performed. The enrichment technology of CA and CI in C. axillare was optimized using D101 macroporous adsorption resin as adsorbent， total contents of CA and CI as evaluation indexes， with the volume fraction and dosage of eluent for impurities and target components. The validation test was also performed. RESULTS The optimum extraction conditions of CA and CI from C. axillare were as follows： the medicinal powder of C. axillare was extracted by ultrasound at room temperature for 45 min at one time with 8 times of 50% ethanol. Results of validation tests showed that total transfer rate of CA and CI in C. axillare was 95.43% in average （RSD＝1.02%， n＝3）. The optimal enrichment technology was as follows： the sample solution was added into D101 macroporous adsorption resin column and stood for 1 hour； the impurities were eluted with 20% ethanol 4 BV （column volume）， and CA and CI were eluted with 50% ethanol 4 BV. The results of validation tests showed that total content of CA and CI was 322.7 mg/g in average （RSD＝1.05%， n＝3）， with average enrichment multiple of 11.61 times. CONCLUSIONS The study has successfully optimized the extraction and enrichment technology of CA and CI from C. axillare， and can provide reference for the development and utilization of CA and CI.
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Strengthening the training of communication ability between doctors and patients of clinical interns in the department of cardiovascular internal medicine, improving the communication ability between doctors and patients of interns, and reducing the occurrence of medical disputes are the key links in training qualified professionals of cardiovascular medicine. Strengthening the pre-job training of interns can stimulate the enthusiasm of interns to improve the communication ability between doctors and patients. Improving the teaching mode of internship will cultivate the communication ability of students and the application ability of professional knowledge in practice. Guiding students to strengthen the training of communication skills will constantly accumulate the experience and lessons of communication between doctors and patients in clinical practice. Enhancing the management supervision will attach importance to the operation test of communication skills between doctors and patients. Using new media technology will establish a platform for communication and information sharing between doctors and patients. Through the above measures, better clinical teaching effect can be achieved in the process of strengthening the communication ability between doctors and patients.
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Objective:To develop a symptom assessment scale after oral thyroidectomy, and to test the reliability and validity of the scale.Methods:On the basis of sufficient literature review in the early stage, Delphi expert correspondence and other methods were used to compile the pre-survey scale for the symptom assessment scale after oral thyroidectomy. The formed pre-survey scale was tested and modified, and then 268 patients were selected for formal survey, and factor analysis was used to form the final version of the scale.Results:After exploratory factors, four common factors were formed, namely, physiological state, cognitive state, complication state, and rehabilitation state. There were 24 items in the scale, and the cumulative variance contribution rate was 67.362%. The Cronbach′s α coefficient was 0.923, the Spearman-Brown splitting reliability of the total scale was 0.914, and the Spearman-Brown splitting reliability of each dimension were 0.987, 0.891, 0.941, 0.936, respectively. The correlation coefficients between the patient′s self-perceived calibration and the total score of the scale and the scores of each dimension were 0.792, 0.567, 0.518, 0.551, 0.535, respectively. The correlation coefficients between the total score of the scale and the four dimensions were 0.562-0.714.Conclusion:The Symptom Evaluation Scale for Oral Thyroidectomy has good reliability and validity, and can be used to evaluate related symptoms after thyroidectomy.
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AMP-activated protein kinase (AMPK) is an energy sensor, which can regulate the energy metabolism of cells by regulating mitochondrial oxidation capacity and substrate utilization, affecting various pathways such as glucose and fatty acid transport. Recent studies have shown that energy metabolism disturbance is a "blasting fuse" leading to a series of secondary injuries after cerebral ischemia-reperfusion, including oxidative stress, acidosis, apoptosis, and excitotoxicity. AMPK participates in most pathophysiological processes of cerebral ischemia-reperfusion injury by regulating cellular energy metabolism. This article reviews the mechanism of action of AMPK signaling pathway in cerebral ischemia-reperfusion injury.
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ObjectiveTo determine the influence of Buzhong Yiqitang on miRNA expression in thyroid tissues of mice with autoimmune thyroiditis （AIT）. MethodThirty female 8-week-old NOD.H-2h4 mice were randomly assigned into normal control group， model group， and Buzhong Yiqitang group （BG）， 10 in each group. Mice were subjected to a diet containing 0.05% sodium iodide for 8 weeks to build the AIT mouse model. After 8 weeks of administration （ig）， samples were collected. A thyroid biopsy was performed on each group of mice， and differential miRNAs in thyroid tissues from each group of mice were analyzed based on experimental validation and bioinformatics. ResultCompared with the conditions of normal control group， thyroid lymphocytes had significant inflammatory infiltration， and there was an increase in serum TgAb level and interleukin（IL）-6 and IL-17 expression and a decrease in IL-1β expression in mice of the model group （P<0.05， P<0.01）. In addition， 154 differentially expressed miRNAs were found. Compared with the conditions of model group， the degree of thyroid tissue inflammation was alleviated， and serum TgAb level， and IL-1β， IL-6 and IL-17 expression of mice treated with the Buzhong Yiqitang were reduced （P<0.05， P<0.01）. Additionally， 112 differentially expressed miRNAs were identified in the BG group. Validation using real-time polymerase chain reaction（Real-time PCR） showed the same trend for miR-326-3p， miR-128-3p， miR-223-5p， miR-141-3p， miR-871-3p， and miR-204-3p as that obtained from miRNA sequencing. In particular， gene ontology（GO） functions were enriched for regulation of T cell activation， oxidative stress， and miRNA binding. Pathways identified by Kyoto encyclopedia of genes and genomes（KEGG）database tended to be enriched in phosphatidylinositol 3-kinases（PI3K）/protein kinase B（Akt）， mitogen-activated protein kinase（MAPK）， and cyclic adenosine monophosphate（cAMP） signaling pathways. Based on miRNA prediction differences， three key genes were identified： SMAD3， JAK2， and STAT3. ConclusionBushong Yiqitang might treat autoimmune thyroiditis by regulating 6 miRNAs.
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From April 2017 to June 2020, 46 patients with acute non-variceal upper gastrointestinal bleeding (ANVUGIB) underwent endoscopic vascular embolization (EVE) in the Department of Gastroenterology of the First Affiliated Hospital of University of Science and Technology of China for rebleeding after endoscopic hemostasis therapy (including local drug injection, electrocoagulation, hemostatic clamp and ligation, etc.). All 46 patients immediately stopped bleeding after EVE, and the effective rate of immediate hemostasis was 100.0%. Postoperative abdominal pain occurred in 13 cases (28.3%), abdominal distension in 3 cases (6.5%) and fever in 2 cases (4.3%). The mucosa healed gradually under gastroscopy 3 and 12 months after the operation. No gastrointestinal rebleeding occurred during the follow-up. Therefore, EVE is a safe and effective method for ANVUGIB patients with failure of initial endoscopic hemostasis, which is worthy of further clinical study and application.
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Objective:To construct a mutant strain of hypervirulent Klebsiella pneumoniae NTHU-K2044 with hfq gene deletion and to analyze its biological characteristics. Methods:The hfq gene of NTUH-K2044 was knocked out by homologous recombination technology to construct △ hfq mutant strain. Its biological characteristics including growth rate, environmental stress tolerance, biofilm formation, capsular polysaccharide synthesis, resistance to neutrophil phagocytosis and lethality to Galleria mellonella larvae were analyzed by comparing with the wild-type strain using phenotypic experiments. Results:The △ hfq mutant strain of hypervirulent Klebsiella pneumoniae NTHU-K2044 was successfully constructed. Phenotypic experiments showed that the △ hfq mutant strain had significantly slower growth rate, smaller colonies and decreased hypermucoviscosity. Its growth was significantly inhibited under different environmental stress conditions such as pH9, pH5.5, 0.7 mmol/L SDS, 5% NaCl, 0.1% H 2O 2 and high temperature of 50℃. In terms of virulence and pathogenicity, the △ hfq mutant strain showed decreased ability to form biofilm and capsule, significantly down-regulated expression of magA and rmpA genes required for capsule synthesis, lower survival rate in the neutrophil bactericidal test and obviously reduced lethality to Galleria mellonella larvae. Conclusions:As a RNA chaperone, Hfq protein could participate in post-transcriptional regulation and play an important role in regulating the physiology, environmental adaptability and virulence of hypervirulent Klebsiella pneumoniae. This study provided reference for further study on hypervirulent Klebsiella pneumoniae.
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Objective:To study the clinical characteristics of purulent meningitis complicated with hydrocephalus in neonates, and to analyze the risk factors of the disease.Methods:Neonates diagnosed with purulent meningitis complicated with hydrocephalus who hospitalized in the department of neonatology of the Second Affiliated Hospital of Wenzhou Medical University from January 2002 to August 2021 were selected as the case group. Neonates with positive pathogen cultures but no hydrocephalus during the same period were assigned by random number table method as the control group. The ratio of the control group and the case group was 2 ∶1. The clinical data such as bacteria distribution, cranial imaging, therapy and prognosis were compared between the two groups. The risk factors for hydrocephalus were predicted. Statistical analysis was conducted using chi-square test and multiple logistic regression analysis.Results:There were 33 cases in the case group and 66 cases in the control group. A total of 27 cases had confirmed pathogen results, of which 20 cases (74.1%) were Gram-negative bacteria and seven cases (25.9%) were Gram-positive bacteria. The time of diagnosis for hydrocephalus were 13.0(5.5, 28.5) days after the onset. Twenty-six cases received non-surgical treatment, while seven cases received surgery. The cure rate of case group was 42.4%(14/33), which was lower than that of control group (72.7%, 48/66), and the difference was statistically significant ( χ2=8.63, P=0.003). Univariate analysis showed that the incidences of protein>3 g/L in cerebrospinal fluid, glucose<2 mmol/L in cerebrospinal fluid, convulsions, central respiratory failure, intracranial hemorrhage and encephalomalacia in the case group were all higher than those in the control group, with statistical significance ( χ2=19.72, 12.04, 19.04, 5.73, 11.85 and 17.48, respectively, all P<0.050). Multivariate logistic regression analysis showed that convulsions (odds ratio ( OR)=4.476, 95% confidence interval ( CI) 1.091 to 18.363, P=0.037), intracranial hemorrhage ( OR=8.031, 95% CI 1.894 to 34.059, P=0.005) and encephalomalacia ( OR=35.189, 95% CI 2.954 to 419.150, P=0.005) were risk factors for neonatal purulent meningitis complicated with hydrocephalus. Conclusions:Gram-negative bacteria are common pathogen of neonatal purulent meningitis complicated with hydrocephalus. Convulsions, intracranial hemorrhage and encephalomalacia are important predictors for neonatal purulent meningitis complicated with hydrocephalus.
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OBJECTIVE@#To explore the correlation of coagulation function with the severity and prognosis of acute pancreatitis (AP) and identify the laboratory markers for early prediction and dynamic monitoring of the prognosis of AP.@*METHODS@#We retrospectively analyzed the clinical data of patients with AP admitted less than 72 h after onset to our hospital from December 1, 2017 to November 30, 2018. The correlation of coagulation function-related markers at admission and their changes during hospitalization with the prognosis of the patients was analyzed.@*RESULTS@#We screened the data of a total of 1260 patients with AP against the inclusion and exclusion criteria, and eventually 175 patients were enrolled in this analysis, among whom 52 patients had severe AP (SAP) and 12 patients died. Logistic regression analysis identified vWF: Ag, PT, PC, AT Ⅲ and D-dimer markers at admission as independent risk factors for predicting SAP and death. Dynamic monitoring of the changes in coagulation function-related markers in the disease course had greater predictive value of the patients' prognosis, and the indicators including vWF: Agmax, PTmax, APTTmax, TTmax, FIBmin, D-dimermax, PLTmin, PCmin, PLGmin, AT Ⅲmin, and their variations were all independent risk factors for predicting SAP and death. ROC analysis suggested that dynamic monitoring of the changes in the indicators, especially those of △vWF: Ag, △PT, △APTT, △FIB, △TT, △D-dimer, △PLT, △PC, △AT Ⅲ, △PLG, could effectively predict SAP and death in these patients (with AUC range of 0.63-0.84).@*CONCLUSION@#Patients with AP have vascular endothelial injuries and coagulation disorders. The markers including vWF: Ag, PT, PC, AT Ⅲ and D-dimer at admission are independent risk factors for predicting SAP and death, and dynamic monitoring of the changes in vWF: Ag、PT、APTT、TT、FIB、D-dimer、PLT、PC、AT Ⅲ and PLG can further increase the predictive value.
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Humans , Acute Disease , Biomarkers , Pancreatitis/diagnosis , Prognosis , ROC Curve , Retrospective Studies , Severity of Illness Index , von Willebrand FactorABSTRACT
Objective:To observe the effect of electroacupuncture based on transcranial magnetic stimulation (TMS) on wakefulness for consciousness disorder. Methods:From January, 2015 to October, 2019, 30 inpatients with consciousness disorder from the General Hospital of Western Theater Command were randomly divided into control group (n = 15) and observetion group (n = 15), Both groups received TMS, and the observetion group received electroacupuncture at Baihui (GV20), Shenting (GV24), bilateral Neiguan (PC6) bilateral Hegu (LI4) and bilateral Yongquan (KI1) in addition, for three months. They were tested the latency of the mismatch negativity (MMN), and assessed with Glasgow Coma Scale (GCS) and persistent vegetative state (PVS) score before, and one month and three months after treatment. Results:The latency of MMN decreased more in the observetion group than in the control group three months after treatment (t = 2.159, P < 0.05). Conclusion:Combination of electroacupuncture on TMS is more effective on wakefulness for consciousness disorder.
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Objective To investigate the influence of Tai Chi posture on knee joint load and the strategy of coordinated muscle contraction. Methods Twenty Tai Chi practitioners with more than 3 years of Tai Chi practice was recruited. By using high-speed motion capture system andthree-dimensional force plate, the adduction angle, tibial angle, joint force, joint torque of the supporting leg during the Tai Chi step lunge movement were collected, and the quadriceps and gastrocnemius muscle strength were obtained through simulation method. The muscle strength performance, recruitment mode and activation method of baseline step, toe-out step and toe-in step were compared. ResultsThe tibial angle and adduction force increased in the toe-out step but those parameters in the toe-in step remained unchanged; the strength of the medial femoris, semimembranosus, semitendinosus and lateral gastrocnemius muscle increased in the toe-out step; the external femoral muscle and medial gastrocnemius muscle increased in the toe-in step. The gastrocnemius muscle was preferentially activated than the quadriceps muscle, and the way of muscle recruitment changed. Conclusions Tai Chi posture will change the way of muscle force recruitment and affect the knee joint function. Standardized movements can be used as the reference for osteoarthritis exercise therapy and have some significance in clinical application.
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Even patients after standard surgery and adjuvant chemotherapy still have a high risk of recurrence and metastasis. With the success of immunotherapy in advanced non-small cell lung cancer (NSCLC), the application of immunotherapy in locally advanced NSCLC has being investigated to reduce the recurrence and metastasis. Pre-clinical studies and several phase II clinical studies had provided theoretical support and clinical evidence for neoadjuvant immunotherapy for NSCLC. This review describes the mechanism of neoadjuvant immuno-chemotherapy, summarizes up-to-date clinical studies, and analyzes efficiency and feasibility of neoadjuvant immune monotherapy or immuno-chemotherapy. Results from four studies (NCT02259621, NEOSTAR, LCMC3 and ChiCTR-OIC-17013726) showed efficiency and feasibility of neoadjuvant anti-programmed cell death 1 (PD-1)/programmed cell death ligand 1 (PD-L1) monotherapy. Neoadjuvant nivolumab plus ipilimumab achieved higher major pathological response rate than nivolumab monotherapy. However, the combination of nivolumab plus ipilimumab led to more severe adverse events as is seen in the NEOSTAR trial. Results from NCT02716038, SAKK 16/14 and NADIM studies suggest that the pathological response rate of neoadjuvant immune-chemotherapy is higher than neoadjuvant immune checkpoint inhibitor monotherapy. This review also elaborates the mechanism of chemotherapy combined with immunotherapy, and discusses the efficacy evaluation after neoadjuvant immunotherapy. .
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Objective:To investigate the clinical characteristics and pathogenic gene mutation of lateral meningocele syndrome(LMS).Methods:We retrospectively collected the clinical manifestations, laboratory examination, imaging examinations, and genetic analysis of a neonate with LMS which was diagnosed at the Department of Neonatology of the Second Affiliated Hospital of Wenzhou Medical University in May 2020. Relevant literature up to February 2021, retrieved from PubMed, OMIM, CNKI, Wanfang, and CQVIP database with the terms of "lateral meningocele syndrome", " NOTCH3", were reviewed to summarize the clinical characteristics, pathogenesis, and genetic etiology of this disease. Results:A full-term male newborn was admitted to our hospital due to feeding difficulty 7 d after birth. The clinical characteristics included hypotonia, dysphagia, hypertension, lateral spinal meningocele, craniofacial anomaly, and cryptorchidism. Abnormal spinal MRI and brainstem evoked potential were also observed. Whole exome sequencing revealed a heterozygous frameshift variation c.6667_6686del(p.Ala2223Profs*12) of NOTCH3 gene located in 19p13.12, which was not detected in the parents. Only 12 English literature were retrieved, with 17 patients from 15 pedigrees. Out of the 18 patients including the index case, 10 were genetically diagnosed as LMS. The age at diagnosis ranged from 15 d to 55 years. Regarding the clinical features, multiple lateral thoracolumbar spinal meningoceles (18/18) was the most common one, followed by retrognathia and low-set ears (16/18), eyelid ptosis and down slanting palpebral fissures (15/18), hypotonia (13/18), hypertension (11/18), developmental delay (9/18), mixed or conductive hearing loss (9/18), cardiovascular dysplasia (7/18), and cryptorchidism (7/10). A total of nine NOTCH3 gene variants were detected, all were heterozygous variants, including six frameshift and three nonsense variants. Conclusions:LMS is caused by NOTCH3 gene mutation with the clinical characteristics including multiple lateral thoracolumbar spinal meningoceles, craniofacial dysmorphisms, hypotonia, hypertension, developmental delay, difficulty in feeding, cryptorchidism, etc.
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Objective:To investigate the role and mechanism of (histone deacetylase 6, HDAC6) in the epithelial-mesenchymal transition (EMT) of renal tubular epithelial cells and the activation of renal interstitial fibroblasts.Methods:Human renal tubular epithelial cells (HK-2) and rat renal interstitial fibroblast (NRK-49F) were cultured in vitro, and divided into 4 groups: control group, Tubastatin A (TA) group (treated with 10 μmol/L HDAC6 inhibitor TA for 36 h), transforming growth factor-β1 (TGF-β1) group (10 ng/ml TGF-β1 for 36 h), and TGF-β1+TA group (treated with 10 ng/ml TGF-β1 and 10 μmol/L TA for 36 h). The expression levels of fibronectin, α-smooth muscle actin (α-SMA), collagen I, E-cadherin, HDAC6, acetyl histone H3, histone H3, acetyl α-tubulin, α-tubulin, TGF-β receptor (TGF-βR) 1, p-Smad3, Smad3, connective tissue growth factor (CTGF), epidermal growth factor receptor (EGFR) and p-EGFR in HK-2 and NRK-49F cell samples were detected by Western blotting, and quantitative analysis was performed according to gray level. Results:(1) In HK-2 cells stimulated by TGF-β1, TA decreased the expression of fibronectin, α-SMA, collagen I, and increased the expression of epithelial cell marker E-cadherin. Meanwhile, TA decreased the expression of HDAC6 and increased the expression levels of acetyl histone H3 and acetyl α-tubulin (all P<0.05). (2) Compared with the TGF-β1 group, the expressions of TGF-βR1, p-Smad3, CTGF and p-EGFR in TGF-β1+TA group were decreased (all P<0.05), while the total protein levels of Smad3 and EGFR were not significantly different (both P>0.05). (3) In NRK-49F cells stimulated by TGF-β1, TA decreased the expressions of fibronectin, α-SMA, collagen I, TGF-βR1 and p-Smad3 (all P<0.05). Conclusions:Blockade of HDAC6 by TA may inhibit the EMT of renal tubular epithelial cells and the activation of renal interstitial fibroblasts via regulating multiple signaling pathways including TGF-β/Smad3, CTGF and EGFR.
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Parathyroid hormone plays an important role in maintaining the homeostasis of calcium and phosphorus metabolism via acting on bone, kidney, and intestine. However, little is known about the effects of parathyroid diseases on the reproductive system. This article describes the association of parathyroid diseases with reproductive function and health.
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Objective:To evaluate the effect of teacher-assisted jaw thrust maneuver on the learning effectiveness of Bonfils fiberscope-guided tracheal intubation for the beginners.Methods:Forty-eight accompanying physicians who were receiving residents standardized training in the Department of Anesthesiology, Peking Union Medical College from April, 2020 to March 2021 and served as trainees were enrolled in this study.The trainees received Bonfils fiberscope training and were randomly divided into independent group ( n=24) and assisted group ( n=24). After induction of general anesthesia, the trainees independently performed the Bonfils fiberscope-guided endotracheal intubation operation in independent group, and trainees performed the operation with the assistance of the teacher′s jaw thrust maneuver in assisted group.The success of intubation at first attempt, duration of intubation, times of teachers′ guidance and scores for trainees′ learning confidence were recorded. Results:Compared with independent group, the success rate of intubation at first attempt was significantly increased, the duration of intubation was shortened, scores for trainees′ learning confidence was increased ( P<0.05), and no significant change was found in the times of teachers′ guidance in assisted group ( P>0.05). Conclusion:For the beginners learning Bonfils fiberscope-guided tracheal intubation, teacher-assisted jaw thrust maneuver is helpful in raising the learning effectiveness and increasing the beginners′ learning confidence.
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Objective:Chemical constituents in hypoglycemic effective fractions of Longan Folium were isolated and identified by ultra performance liquid chromatography-quadrupole/electrostatic field orbitrap high resolution mass spectrometry （UPLC-Q-Orbitrap HRMS） to clarify the hypoglycemic substance basis of Longan Folium. Method:Chemical constituents in hypoglycemic effective fractions of Longan Folium were isolated on a Thermo Hypersil GOLD C<sub>18</sub> column （2.1 mm×100 mm， 1.9 μm）， the mobile phase was 0.1% formic acid acetonitrile solution and 0.1% formic acid solution （containing and 10 mmol ammonium acetate） for gradient elution. HRMS was operated in the positive and negative ion modes with the scanning range of <italic>m</italic>/<italic>z</italic> 100-1 500. Result:The secondary fragment ion information of target compounds was selected and compared with the compounds reported in the databases and related literature to further confirm these compounds. Nine compounds were identified in the ethanol fraction of Longan Folium， including cynaroside， kaempferol， quercitrin， luteolin， shikimic acid， citric acid， <italic>L</italic>-tyrosine， adenosine and nicotinamide. A total of 11 compounds were determined in the ethyl acetate fraction （cynaroside， quercitrin， kaempferol， luteolin， shikimic acid， gallic acid， protocatechuic acid， adenosine， nicotinamide， <italic>L</italic>-phenylalanine and scopoletin）， and 10 compounds were identified in the <italic>n</italic>-butanol fraction （cynaroside， kaempferol-3-<italic>O</italic>-rutinoside， kaempferol， astragalin， luteolin， citric acid， gallic acid， adenosine， nicotinamide and 5-hydroxymethylfurfural）. And five common compounds were identified in these three hypoglycemic effective fractions. Conclusion:The established UPLC-Q-Orbitrap HRMS can quickly identify chemical constituents in three hypoglycemic effective fractions of Longan Folium， their main chemical constituents are flavonoids and their glycosides， organic acids and nitrogen-containing compounds， which provides technical support and scientific evidence for the study on pharmacodynamic material basis and quality control of Longan Folium.
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Objective:To investigate the mechanism of Xiangsha Liujunzi Tang in improving liver lipid deposition in ApoE<sup>-/-</sup> atherosclerotic （AS） mice by affecting long noncoding RNA-HC （Lnc-HC）/microRNA-130b （miR-130b） in the regulation of cholesterol metabolism. Method:Totolly 10 C57BL/6J mice were selected as normal controls， and 30 healthy ApoE<sup>-/-</sup> mice fed with high fat diet for 12 weeks were then randomly divided into the model group， Xiangsha Liujunzi Tang group（19.12 g·kg<sup>-1</sup>·d<sup>-1</sup>） and simvastatin group（2.275 mg·kg<sup>-1</sup>·d<sup>-1</sup>）， with gavage administration for 4 weeks. The serum lipid level of mice was detected by automatic biochemistry analyzer， and the histopathological changes of liver cells were observed by hematoxylin-eosin （HE） staining. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to detect expression of long noncoding RNA-HC， and miR-130b. Real-time PCR and Western blot assay were used to detect gene and protein expression of peroxisome proliferator-activated receptor gamma （PPAR<italic>γ</italic>）， liver X receptor （LXR）， ATP-binding cassette transporters A1 （ABCA1）， ATP-binding cassette transporters G1 （ABCG1）， ATP-binding cassette transporters G5 （ABCG5）， and ATP-binding cassette transporters G8 （ABCG8）. Result:Compared with the normal control group， the mice in the model group showed abnormal blood lipids， larger liver cells， obvious fat vacuoles， significantly increased expression of Lnc-HC， miR-130b in liver， and significantly decreased gene and protein expression of PPAR<italic>γ</italic>， LXR， ABCA1， ABCG1， ABCG5， and ABCG8 in mice liver （<italic>P</italic><0.05，<italic>P</italic><0.01）. Compared with the model group， the abnormal blood lipid levels of the mice in the Xiangsha Liujunzi Tang group and the simvastatin group were improved， and the number of fatty vacuoles of liver cells was significantly reduced， the expression of liver Lnc-HC， miR-130b in Xiangsha Liujunzi Tang group decreased significantly （<italic>P</italic><0.05，<italic>P</italic><0.01）， the gene and protein levels of liver PPAR<italic>γ</italic>， ABCA1， ABCG1， ABCG5， ABCG8 in mice of the Xiangsha Liujunzi Tang group and the simvastatin group showed an upward trend. Among them， the gene and protein expression of LXR protein in the liver of the Xiangsha Liujunzi Tang group was significantly up-regulated （<italic>P</italic><0.05）. Conclusion:Xiangsha Liujunzi Tang may improve the lipid deposition in the liver of ApoE<sup>-/- </sup>AS mice by affecting Lnc-HC/miR-130b to regulate the cholesterol metabolism process mediated by PPAR<italic>γ</italic>， thus playing a role in preventing and treating AS.
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Peptide inhibition of the interactions of the tumor suppressor protein P53 with its negative regulators MDM2 and MDMX activates P53