The pGenesil-1-Beclin1 eukaryotic expression vectors were constructed to establish an SH-SY5Y cell line stably expressing shRNA-Beclin1. The shRNA was connected to pGenesil-1 to construct the recombinant plasmid pGenesil-1-Beclin1, which was transformed into JM109 E.coli. Positive clones were identified by digestion with restriction endonuclease and DNA sequencing. SH-SY5Y cells were cultured by the conventional method. The pGenesil-1-Beclin1 and pGenesil-1 plasmids were transfected into SH-SY5Ycells, and the cells were screened by G418 until the stable G418-resistant monoclonal cells were acquired. Beclin1 mRNA and Beclin1 protein were detected by RT-PCR and Western blot analysis respectively. The results of restriction endonuclease analysis and DNA sequencing confirmed the correct construction of the eukaryotic expression vector pGenesil-1-Beclin1. Two SH-SY5Y transfected cell lines were successfully selected. Compared with the control group, RT-PCR and Western blot showed that the expression of Beclin1 mRNA and protein were down regulated 71.28%±1.45%(P<0.05)and 75.50%±2.63%(P<0.05), respectively. The results indicated that the eukaryotic expression vector pGenesil-1-Beclin1 was successfully constructed and the SH-SY5Y cell lines with inhibited Beclin1 expression were established. It provides a useful cell model for studying the biological function of Beclin1.
ObjectivesTo explore the effect of the deep learning algorithm convolutional neural network (CNN) in screening of randomized controlled trials (RCTs) in Chinese medical literatures.MethodsLiterature with the topic " oral science” published in 2014 were retrieved from CNKI and exported citations containing title and abstract. RCTs screening was conducted by double independent screening, checking and peer discussion. The final results of the citations were used for CNN algorithm model training. After completing the algorithm model training, a prospective comparative trial was organized by searching all literature with the topic "oral science" published in CNKI from January to March 2018 to compare the sensitivity (SEN) and specificity (SPE) of algorithm with manual screening. The initial results of a single screener represented the performance of manual screening, and the final results after peer discussion were used as the gold standard. The best thresholds of algorithm were determined with the receptor operative characteristic (ROC) curve.ResultsA total of 1 246 RCTs and 4 754 non-RCTs were eventually included for training and testing of CNN algorithm model. 249 RCTs and 949 non-RCTs were included in the prospective trial. The SEN and SPE of manual screening were 98.01% and 98.82%. For the algorithm model, the SEN of RCTs screening decreased with the increase of threshold value while the SPE increased with the increase of threshold value. After 27 changes of threshold value, ROC curve were obtained. The area under the ROC curve was 0.9977, unveiling the optimal accuracy threshold (Threshold=0.4, SEN=98.39%, SPE=98.84%) and high sensitivity threshold (Threshold=0.06, SEN=99.60%, SPE=94.10%).ConclusionsA CNN algorithm model is trained with Chinese RCTs classification database established in this study and shows an excellent classification performance in screening RCTs of Chinese medical literature, which is proved to be comparable to the manual screening performance in the prospective controlled trial.
Abstract： Pulmonary metastasectomy is an option for patients with metastatic tumor of lung. Numerous retrospectivestudies have demonstrated that complete control of primary tumor and complete resection of metastases limited to thelungs may be associated with prolonged survival. Speci?c issues require consideration when planning pulmonary metastasectomy. Regardless of histological type of primary tumor, complete resection is the most important prognostic factor. The other two important prognostic factors are long disease interval and limited number of metastatic tumor of lung. Hand-assisted thoracoscopic surgery for bilateral lung metastasectomy through sternocostal triangle access is recommended. Pulmonary hilar and mediastinal lymph node metastases are some relative contraindications for this surgery. Nowdays preoperative imaging examinations still have limitations in detecting all the lung metastases. Some data emphasize the importance of considering patients for extended resection in metastatic tumor of lung. Repeat resection after previous metastasectomies can be of benefit under certain circumstances so we should remove as little healthy lung tissue as possible. In this review, we discuss about some disputed issues in order to establish a useful criterion for consideration of pulmonary metastasectomy.
OBJECTIVE: To investigate the selection and identification of human keratinocyte stem cells(KSC) in vitro. METHODS: According to the characteristics of KSC which can adhere to extracellular matrix very fast, we selected 3 groups of different time(5 minutes, 20 minutes and 60 minutes) and unselected as control group. And the cells were identified by monoclone antibody of beta 1-integrin and cytokeratin 19 (Ck19), then the image analysis was done. Furthermore we analyzed the cultured cells with flow cytometer(FCM) and observed the ultrastructure of the cell by transmission electron microscope(TEM). RESULTS: The cell clones formed in all groups after 10 to 14 days, while the cells of 5 minute group grew more slowly than those of the other groups, however, the clones of this group were bigger. The expression of beta 1-integrin and Ck19 were found in all groups. The positive rate of beta 1-integrin was significant difference between 5 minute group and the other groups (P lt; 0.05). And the expression of Ck19 was no significant difference between 5 minute group and 20 minute group(P gt; 0.05), and between 60 minute group and control group. But significant difference was observed between the former and the later groups(P lt; 0.05). The result of FCM showed that most cells of the 5 minute group lied in G1 period of cell cycle, which was different from those of the other groups. At the same time, the cells of 5 minute group were smaller and contained fewer organelles than those of the other groups. CONCLUSION: The above results demonstrate that the cells of 5 minute group have a slow cell cycle, characteristics of immaturity, and behaving like clonogenic cells in vitro. The cells have the general anticipated properties for KSC. So the KSC can be selected by rapid attachment to extracellular matrix and identified by monoclone antibody of beta 1-integrin and Ck19.
Hypertension is the primary disease that endangers human health. A convenient and accurate blood pressure measurement method can help to prevent the hypertension. This paper proposed a continuous blood pressure measurement method based on facial video signal. Firstly, color distortion filtering and independent component analysis were used to extract the video pulse wave of the region of interest in the facial video signal, and the multi-dimensional feature extraction of the pulse wave was preformed based on the time-frequency domain and physiological principles; Secondly, an integrated feature selection method was designed to extract the universal optimal feature subset; After that, we compared the single person blood pressure measurement models established by Elman neural network based on particle swarm optimization, support vector machine (SVM) and deep belief network; Finally, we used SVM algorithm to build a general blood pressure prediction model, which was compared and evaluated with the real blood pressure value. The experimental results showed that the blood pressure measurement results based on facial video were in good agreement with the standard blood pressure values. Comparing the estimated blood pressure from the video with standard blood pressure value, the mean absolute error (MAE) of systolic blood pressure was 4.9 mm Hg with a standard deviation (STD) of 5.9 mm Hg, and the MAE of diastolic blood pressure was 4.6 mm Hg with a STD of 5.0 mm Hg, which met the AAMI standards. The non-contact blood pressure measurement method based on video stream proposed in this paper can be used for blood pressure measurement.
Systematic reviews can provide important evidence support for clinical practice and health decision-making. In this process, literature screening and data extraction are extensively time-consuming procedures. Natural language processing (NLP), as one of the research directions of computer science and artificial intelligence, can accelerate the process of literature screening and data extraction in systematic reviews. This paper introduced the requirements of systematic reviews for rapid literature screening and data extraction, the development of NLP and types of machine learning; and systematically collated the NLP tools for the title and abstract screening, full-text screening and data extraction in systematic reviews; and discussed the problems in the application of NLP tools in the field of systematic reviews and proposed a prospect for its future development.
The emergence of regular short repetitive palindromic sequence clusters (CRISPR) and CRISPR- associated proteins 9 (Cas9) gene editing technology has greatly promoted the wide application of genetically modified pigs. Efficient single guide RNA (sgRNA) is the key to the success of gene editing using CRISPR/Cas9 technology. For large animals with a long reproductive cycle, such as pigs, it is necessary to screen out efficient sgRNA in vitro to avoid wasting time and resource costs before animal experiments. In addition, how to efficiently obtain positive gene editing monoclonal cells is a difficult problem to be solved. In this study, a rapid sgRNA screening method targeting the pig genome was established and we rapidly obtained Fah gene edited cells, laying a foundation for the subsequent production of Fah knockout pigs as human hepatocyte bioreactor. At the same time, the method of obtaining monoclonal cells using pattern microarray culture technology was explored.
随着人口的老龄化，越来越多的有症状或无症状的冠心病患者需接受非心脏外科手术。接受非心脏外科手术而死亡的患者大约有50%是由于心脏并发症所致［1］。围手术期发生的心脏并发症大约5%～10%为心肌梗死，主要发生于术后头3天，其病死率很高，可达32%～69%［2，3］。术后发生心肌梗死或不稳定型心绞痛的患者发生心血管问题的几率增加20倍［4］。因此，如何评估非心脏外科手术患者的心脏危险性，如何预防围手术期心脏并发症的发生，已成为外科医生十分关注的一个问题。
We aim to screen out the active components that may have therapeutic effect on coronavirus disease 2019 (COVID-19) from the severe and critical cases’ prescriptions in the “Coronavirus Disease 2019 Diagnosis and Treatment Plan (Trial Ninth Edition)” issued by the National Health Commission of the People’s Republic of China and explain its mechanism through the interactions with proteins. The ETCM database and SwissADME database were used to screen the active components contained in 25 traditional Chinese medicines in 3 prescriptions, and the PDB database was used to obtain the crystal structures of 4 proteins of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Molecular docking was performed using Autodock Vina and molecular dynamics simulations were performed using GROMACS. Binding energy results showed that 44 active ingredients including xambioona, gancaonin L, cynaroside, and baicalin showed good binding affinity with multiple targets of SARS-CoV-2, while molecular dynamics simulations analysis showed that xambioona bound more tightly to the nucleocapsid protein of SARS-CoV-2 and exerted a potent inhibitory effect. Modern technical methods are used to study the active components of traditional Chinese medicine and show that xambioona is an effective inhibitor of SARS-CoV-2 nucleocapsid protein, which provides a theoretical basis for the development of new anti-SARS-CoV-2 drugs and their treatment methods.