OBJECTIVES: To investigate the effect of BRD4 inhibition on migration of esophageal squamous cell carcinoma (ESCC) cells with low acetyl-CoA carboxylase 1 (ACC1) expression. METHODS: ESCC cell lines with lentivirus-mediated ACC1 knockdown or transfected with a negative control sequence (shNC) were treated with DMSO, JQ1 (a BRD4 inhibitor), co-transfection with shNC-siBRD4 or siNC with additional DMSO or C646 (an ahistone acetyltransferase inhibitor) treatment, or JQ1combined with 3-MA (an autophagy inhibitor). BRD4 mRNA expression in the cells was detected using RT-qPCR. The changes in cell proliferation, migration, autophagy, and epithelial-mesenchymal transition (EMT) were examined with CCK8 assay, Transwell migration assay, and Western blotting. RESULTS: ACC1 knockdown did not significantly affect BRD4 expression in the cells but obviously increased their sensitivity to JQ1. JQ1 treatment at 1 and 2 μmol/L significantly inhibited ESCC cell proliferation, while JQ1 at 0.2 and 2 μmol/L promoted cell migration. The cells with ACC1 knockdown and JQ1 treatment showed increased expresisons of vimentin and Slug and decreased expression of E-cadherin. BRD4 knockdown promoted migration of ESCC cells, and co-transfection with shACC1 and siBRD4 resulted in increased vimentin and Slug expressions and decreased E-cadherin expression in the cells. C646 treatment of the co-transfected cells reduced acetylation levels, decreased vimentin and Slug expressions, and increased E-cadherin expression. Treatment with JQ1 alone obviously increased LC3A/B-II levels in the cells either with or without ACC1 knockdown. In the cells with ACC1 knockdown and JQ1 treatment, additional 3-MA treatment significantly decreased the expressions of vimentin, Slug and LC3A/B-II and increased the expression of E-cadherin. CONCLUSIONS BRD4 inhibition promotes autophagy of ESCC cells via a histone acetylation-dependent mechanism, thereby enhancing EMT and ultimately increasing cell migration driven by ACC1 deficiency.
Humans ; Cell Movement ; Transcription Factors/metabolism* ; Esophageal Neoplasms/metabolism* ; Cell Line, Tumor ; Cell Cycle Proteins ; Azepines/pharmacology* ; Epithelial-Mesenchymal Transition ; Carcinoma, Squamous Cell/metabolism* ; Esophageal Squamous Cell Carcinoma ; Triazoles/pharmacology* ; Nuclear Proteins/genetics* ; Cell Proliferation ; Acetyl-CoA Carboxylase/genetics* ; Transfection ; Autophagy ; Bromodomain Containing Proteins

In-depth study of the components of polymyxins is the key to controlling the quality of this class of antibiotics. Similarities and variations of components present significant analytical challenges. A two-dimensional (2D) liquid chromatography-mass spectrometr (LC-MS) method was established for screening and comprehensive profiling of compositions of the antibiotic colistimethate sodium (CMS). A high concentration of phosphate buffer mobile phase was used in the first-dimensional LC system to get the components well separated. For efficient and high-accuracy screening of CMS, a targeted method based on a self-constructed high resolution (HR) mass spectrum database of CMS components was established. The database was built based on the commercial MassHunter Personal Compound Database and Library (PCDL) software and its accuracy of the compound matching result was verified with six known components before being applied to genuine sample screening. On this basis, the unknown peaks in the CMS chromatograms were deduced and assigned. The molecular formula, group composition, and origins of a total of 99 compounds, of which the combined area percentage accounted for more than 95% of CMS components, were deduced by this 2D-LC-MS method combined with the MassHunter PCDL. This profiling method was highly efficient and could distinguish hundreds of components within 3 h, providing reliable results for quality control of this kind of complex drugs.

Cancer is a serious global health problem and a major cause of human death. Conventional cancer treatments often run the risk of impairing vital organ functions. Anticancer peptides (ACPs) are considered to be one of the most promising therapeutic agents against common human cancers due to their small sizes, high specificity, and low toxicity. Since ACP recognition is highly limited to the laboratory, expensive, and time-consuming, we proposed pLM4ACP, a model for predicting ACPs based on machine learning and protein language models. In this model, the protein language model ProtT5 was used to extract the features of ACPs, and the extracted features were input into the support vector machine (SVM) classification algorithm for optimization and performance evaluation. The model showcased significantly higher accuracy than other methods, with the overall accuracy of 0.763, F1-score of 0.767, Matthews correlation coefficient of 0.527, and area under the curve of 0.827 on the independent test set. This study constructs an efficient anticancer peptide prediction model based on protein language models, further advancing the application of artificial intelligence in the biomedical field and promoting the development of precision medicine and computational biology.
Machine Learning ; Antineoplastic Agents/chemistry* ; Humans ; Peptides/chemistry* ; Support Vector Machine ; Algorithms ; Computational Biology/methods* ; Neoplasms/drug therapy*

ObjectiveTo investigate the possible mechanism of Huatan Sanjie Formula （化痰散结方， HSF） in treating Graves' disease （GD） from the perspective of thyroid angiogenesis. MethodsThirty-six BALB/c female mice were randomly divided into a normal control group （n=9） and a modeling group （n=27）. Mice in the modeling group were injected with 2.0×10⁹ PFU/ml of Ad-TSHR289 adenovirus into the tibialis anterior muscle to build GD model. Nine weeks after immunization， the successfully modeled mice were randomly divided into model group， methimazole （MMI） group and HSF group， with 9 mice in each group. The MMI group was given 5.2 mg/（kg·d） of methimazole tablets by gavage， while the HSF group was given HSF at a relative crude drug dosage of 7.02 g/（kg·d） by gavage. The normal control group and the model group were given 0.1 ml/10 g of pure water by gavage. All groups were administered intragastrically once a day for a total of 4 weeks. The levels of thyroxine （T4） and thyrotropin receptor autoantibodies （TRAb） in serum were detected by radioimmunoassay， while the pathological changes of the thyroid gland were assessed by HE staining. The vascular morphology of thyroid tissue was observed by CD34 immunohistochemical staining， and the microvessel density （MVD） was counted. The protein expression of vascular endothelial growth factor A （VEGFA） and vascular endothelial growth factor receptor 2 （VEGFR2） in thyroid was detected by Western-blot. ResultsCompared to those in the normal control group， the thyroid volume of the mice in the model group significantly increased with excessive congestion， and the pathology showed significant thyroid follicular hyperplasia， columnar and proliferated epithelial cells， and enlarged follicle size； serum T4 and TRAb significantly increased， as well as the count of thyroid MVD， and the protein expressions of thyroid VEGFA and VEGFR2 （P＜0.01）. Compared to those in the model group， the thyroid glands of the mice in the MMI group and the HSF group were significantly reduced， and the congestion was improved； pathology showed that thyroid follicular hyperplasia and epithelial cell proliferation were reduced， with smooth edges of the follicles and the significantly reduced inward protrusion； serum T4 and TRAb significantly decreased， as well as the thyroid MVD， thyroid VEGFA and VEGFR2 protein expressions （P＜0.05 or P＜0.01）. There was no significant difference in all indicators between the MMI group and the HSF group （P＞0.05）. ConclusionHSF may inhibit thyroid angiogenesis by down-regulating thyroid VEGFA/VEGFR2 signaling pathway， thereby improving goitre and hyperfunction in GD mice.

Objective:To analyze the functional connectivity (FC) characteristics of sensory motor network (SMN) in patients with bipolar disorder type Ⅰ (BD-Ⅰ) by independent component analysis (ICA), and explore the correlation between abnormal SMN and clinical symptoms.Methods:Eighteen patients with BD-Ⅰ (BD-Ⅰ group) and 20 matched normal controls (HC group) were included.Both groups received resting state fMRI (rs-fMRI) scanning.Based on ICA-fMRI data, one-sample t-test and two-sample t-test were used to analyze the components of SMN and to explore abnormal brain regions between the two groups.Functional network analysis (FNC) was also used to explore the functional connectivity between SMN and other brain networks.Pearson correlation analysis were conducted by SPSS 17.0 to measure the potential associations between intra-and inter-network functional connectivity and age, education, score of Bech-Rafaelsen mania rating scale (BRMS), score of positive and negative syndrome scale (PANSS) and other indicators. Results:In BD-Ⅰ group, the functional connection in the right paracentral lobule (MIN: x=8, y=-32, z=68, t=4.86, P<0.001) and the right postcentral gyrus (MIN: x=41, y=-26, z=53, t=3.33, P<0.001) in SMN were higher than those in HC group.Compared with HC group, the connectivity value in patients with BD-Ⅰ increased between SMN-DAN (0.247±0.073, -0.078±0.080, t=-2.974, P<0.01, FDR adjusted), while the connectivity value decreased between SMN-DMN(-0.037±0.054, 0.272±0.067, t=3.520, P<0.01, FDR adjusted) and between SMN-rFPN(-0.034±0.055, 0.231±0.070, t=2.939, P<0.01, FDR adjusted). Conclusion:The sensorimotor network of patients with BD-Ⅰ has abnormal functional connections within and between networks, and FC values in some networks are positively correlated with manic symptoms, which may be part of the neural mechanisms of patients with BD-Ⅰ.

Objective:To analyze the clinical features of patients with severe dengue (SD) in Guangdong Province, and to improve the understanding of the diagnosis and treatment of SD in China.Methods:The clinical data, laboratory examination and etiological test results of 257 SD cases from 29 dengue fever designated hospitals in Guangdong Province from January 1, 2013 to December 31, 2019 were respectively collected. The relevant indicators of the criteria for severe organ involvement were quantified. Logistic regression analysis was performed to analyze the risk factors for the development of multiple organ failure in SD patients.Results:Among the 257 SD patients, age was (64.1±20.1) years old, with 65.4%(168/257) of them ≥60 years old, 142 were male and 115 were female. One hundred and fifty-two (59.1%) patients had underlying conditions, including 115(44.7%) patients with hypertension. The clinical manifestations were mainly fever (98.4%(253/257)), fatigue (70.0%(180/257)), cough or expectoration (44.4%(114/257)), lethargy or irritability (39.3%(101/257)), vomiting (30.4%(78/257)), abdominal pain or tenderness (20.6%(53/257)), hepatomegaly (2.3%(6/257)), bleeding tendency (59.5%(153/257)), and pleural effusion or ascites (43.6%(112/257)). Platelet count levels were decreased in 90.9%(231/254) of the cases, and 97.1%(234/241) of patients had normal or decreased hematocrit. The most common of severe manifestations were severe organ involvement (61.1%(157/257)), followed by severe bleeding (37.0%(95/257)) and severe plasma leakage (30.0%(77/257)). Severe organ involvements were more common in the kidney (27.6%(71/257)) and heart (26.8%(69/257)). Multivariate logistic regression analysis showed that age (odds ratio ( OR)=1.051, 95% confidence interval ( CI) 1.004 to 1.100, P=0.035), hypertension ( OR=5.224, 95% CI 1.272 to 21.462, P=0.022), elevated aspartate aminotransferase (AST) level ( OR=1.002, 95% CI 1.001 to 1.003, P=0.001), blood urea nitrogen (BUN) ( OR=1.050, 95% CI 1.005 to 1.098, P=0.030), and international normalized ratio (INR) ( OR=4.604, 95% CI 1.601 to 13.238, P=0.005) were risk factors for the development of multiple organ failure in SD patients. The detection results of serum samples form 113 SD patients in acute phase showed that dengue virus (DENV)-1 accounted for 89.4%(101/113), DENV-2 accounted for 9.7%(11/113), and DENV-3 accounted for 0.9% (1/113). Conclusions:Elderly and those with co-existing conditions such as hypertension in SD patients in Guangdong Province are more common. Severe organ involvement such as kidney and heart is the main cause of SD. DENV-1 infection is predominant. Significant elevated levels of AST, BUN and INR may be related to a poor prognosis.

Objective:To evaluate the effect of electroacupuncture postconditioning on the expression of microRNA-1 (miRNA-1) in rats with arrhythmias induced by myocardial ischemia-reperfusion (I/R).Methods:Twenty-four clean-grade healthy adult male Sprague-Dawley rats, aged 2-3 months, weighing 280-320 g, were divided into 3 groups ( n=8 each) using a random number table method: sham operation group (S group), I/R group and electroacupuncture postconditioning group (EP group). The model of myocardial reperfusion arrhythmia was established by ligating the left anterior descending branch of coronary artery for 30 min followed by 30-min reperfusion in anesthetized rats.The artery was only isolated without ligation after thoracotomy in group S. Bilateral Neiguan acupoints were stimulated for 30 min starting from the time point immediately after reperfusion in EP group.The occurrence of arrhythmia during reperfusion was recorded.The rats were sacrificed at 30 min of reperfusion, and hearts were removed for examination of the pathological changes of the myocardium and for determination of miRNA-1 expression (by quantitative real-time polymerase chain reaction) and expression of Kir2.1 and connexin 43 (Cx43) (by Western blot). Results:Compared with S group, the incidence of arrhythmia was significantly increased, the expression of miRNA-1 was up-regulated, and the expression of Kir2.1 and Cx43 was down-regulated in I/R and EP groups ( P<0.05). Compared with I/R group, the incidence of arrhythmia was significantly decreased, the expression of miRNA-1 was down-regulated, and the expression of Kir2.1 and Cx43 was up-regulated in EP group ( P<0.05). The pathological changes of myocardium were accentuated in I/R and EP groups when compared with S group, and the pathological changes were significantly attenuated in EP group when compared with I/R group. Conclusion:The mechanism by which electroacupuncture postconditioning reduces the occurrence of reperfusion arrhythmia is related to up-regulation of Kir2.1 and Cx43 expression after down-regulation of miRNA-1 expression in rats.

Objective:To evaluate the effect of electroacupuncture (EA) on electrophysiological characteristics of ventricular myocardium during myocardial ischemia-reperfusion (I/R) in rats.Methods:Twenty-four clean-grade healthy adult male Sprague-Dawley rats, weighing 280-320 g, were divided into 3 groups ( n=8 each) using a random number table method: sham operation group (group SH), I/R group and group EA.The model of myocardial I/R injury was established by ligating the left anterior descending branch of coronary artery for 30 min followed by 30-min reperfusion in anesthetized rats.Bilateral Neiguan acupoints in forelimbs were stimulated for 30 min during the period of reperfusion in group EA.Heart rate (HR), monophasic action potential amplitude (MAPA), maximum depolarization rate (V max), and monophasic action potential duration at 90% repolarization (MAPD 90) were recorded.The development of arrhythmias and arrhythmias score were recorded during reperfusion. Results:Compared with group SH, HR was significantly decreased, MAPA and V max were decreased, MAPD 90 was prolonged, and the incidence of ventricular premature beat, ventricular tachycardia and ventricular fibrillation and arrhythmias score were increased at T 1, 2 in I/R and EA groups ( P<0.05). Compared with group I/R, HR was significantly increased, MAPA and V max were increased, MAPD 90 was shortened, and the incidence of ventricular tachycardia and ventricular fibrillation and arrhythmias score were decreased at T 2 in EA group ( P<0.05). Conclusion:EA can accelerate myocardial depolarization and shorten repolarization, thus decreasing the occurrence of reperfusion arrhythmia in rats.

Objective: To compare the difference of cognitive fusion, empirical avoidance between patients with obsessive-compulsive disorder and healthy control group, and to explore the relationship between obsessive-compulsive score and cognitive fusion, empirical avoidance. Methods: The cognitive fusion questionnaire(CFQ), acceptance and action questionnaire-2nd edition(AAQ-II) and Yale-Brown scale for obsessive-compulsive symptoms (Y-BOCS) were used to investigate 100 subjects with obsessive-compulsive disorder (OCD) and 166 healthy controls.And the differences in cognitive fusion and empirical avoidance scores were compared between the two groups.The relationship of obsessive-compulsive symptoms and cognitive fusion, empirical avoidance was analyzed using multiple linear regression analysis. Results: The scores of cognitive fusion ((49.89±10.62) vs (33.88±11.44), t=-11.345, P<0.01)and empirical avoidance ((29.75±9.53) vs (21.59±7.03), t=-7.995, P<0.01) in obsessive-compulsive disorder group were significantly higher than those in healthy control group.There were significant differences in cognitive fusion and empirical avoidance in age variables (F=8.63, P<0.01). In AAQ-Ⅱ, item 2 (r=0.246, P<0.05), item 6 (r=0.223, P<0.05) and total score (r=0.240, P<0.05) were positively correlated with the total score of Y-BOCS.Item 2 (r=0.311, P<0.01), item 3 (r=0.286, P<0.05), item 6 (r=0.248, P<0.05) and total score (r=0.229, P<0.05) were positively correlated with the scores of obsessive-thinking.In CFQ, item 2 (r=0.231, P<0.0) 5), item 4 (r=0.242, P<0.05), item 7 (r=0.308, P<0.05), item 8 (r=0.277, P<0.05) and item 9 (r=0.249, P<0.05) were positively correlated with the total score of Y-BOCS.Item 8 (r=0.261, P<0.05) was positively correlated with the scores of obsessive-thinking, item 7 (r=0.237, P<0.05) and item 9 (r=0.238, P<0.05) were positively correlated with scores of obsessive-compulsive behavior.When predicting total obsessive-compulsive scores, only CF item 7 of Q (B=1.827, P<0.01), item 3 (B=0.956, P<0.05), and item 6 of AAQ-Ⅱ (B=0.584, P<0.05) entered the equation with a joint explanatory variation of 19%.When predicting the score of obsessive-thinking, only item 2 of AAQ-Ⅱ (B=0.446, P<0.01) entered the equation, explaining the variance was 9.7%.In the prediction of obsessive-compulsive behavior, only item 9 (B=0.815, P<0.05) of CFQ entered the equation, and the explanatory variation was 5.6%. Conclusion Cognitive fusion in the patients with obsessive-compulsive disorder and the high level of empirical avoidance may be an important factor for the maintenance of the symptoms.

Objective This study was to study the effect of monovision contact lens (MV-CL) on distant and near vision,distant and near stereopsis,and to identify the effect of degree of monovision on vision function.Methods A prospective case observational study was designed.The clinical data of sixty-eight eyes of 47 cases were collected from November 2010 to August 2014 in Henan Provincial Corps Hospital,Chinese People's Armed Police Forces.Sixty-eight eyes of 47 cases wore contact lenses to improve near vision,in which the other eye of 21 cases with hyperopia wearing contact lenses to improve distant vision.Besides,far and near stereopsis were measured before and two weeks after wearing lenses respectively;and scotopia were measrued with lenses and in naked eyes respectively.Compared the changes of vision and stereopsis in patients before and after the MV-CL,and conducted the patient satisfaction survey.Results There were 89％ (42/47 cases) and 6％ (3/47 cases) of the patients whose naked far and near vision was 0.8/0.33 (J4) at 2 weeks after MV-CL and before MV-CL,respectively.There were no statistical significance about change of near stereopsis with Titmus stereopicture and the Yan's near random-dot stereogram (x2=1.30,0.56;both at P＞0.05).There were no statistical significance about change with apparent machine random dot stereogram iterative crossed parallax and uncrossed disparity (x2 =0.16,0.11;both at P＞0.05).The far distance fusion range and far stereopsis were not significantly different between before and after MV-CL (x2 =0.22,0.16;both at P＞0.05).Titmus stereopicture tests showed that there was a decrease in the number of people who had foveal stereopsis,while an increase was found in people who had macular stereopsis after MV-CL,but the difference had no statistical significance (x2 =2.28,P ＞ 0.05).Conclusions Near vision can be obviously increased and presbytism can be relieved by MV-CL.Stereopsis of some patients are decreased after MV-CL,but this can be accepted by the patients and there is no obvious decrease in far range of fusion.