Phage display technology is a kind of laboratory technique by means of genetic engineering to combine phage with exogenous genes, based on the presentation of a heterogeneous peptide or protein libraries on the surface of phage along with phage coat protein expressions. The displayed proteins or peptides could maintain their own relative spatial structure and biological activity, allowing them to direct phage binding to a target of interest. Therefore, the technology can be widely used as a simple and effective screening tool to screen successfully tumor-targeted peptides, deliver tumor-targeted drugs and prepare tumor antibody, which plays an important role in tumor-targeted therapy.

IFIT family genes are a kind of interferon stimulated genes (ISGs), and play important roles in antiviral sector and immunity regulation. To study the regulatory effect of IFIT family genes during influenza A virus (IAV) infection, we used RNA-sequencing analysis (RNA-Seq) technique and found that when 293T cells were infected by A/WSN/33 (WSN), the concentration of IFIT family genes were increased. Further study reveals that overexpression of IFIT2 or IFIT3 could inhibit IAV replication and transcription, and cause the dose-dependent inhibition of polymerase activity of vRNP. In addition, IFIT2 and IFIT3 encoding protein could colocalize with NS1 in 293T cells infected by WSN, indicating that they might interact with each other. The results suggest that IFIT family genes can inhibit the replication and transcription of IAV, which contributes to our understanding of the regulatory effect of host factors during influenza virus infection.
HEK293 Cells ; Humans ; Influenza A virus ; physiology ; Influenza, Human ; genetics ; Intracellular Signaling Peptides and Proteins ; genetics ; Proteins ; genetics ; Virus Replication

Objective To understand the clinical features of Behcet′s Disease( BD) with pulmonary hypertension( PH) .Methods The etiology, clinical features, treatment and prognosis of 25 patients with Bahcet′s disease ( BD) complicated with pulmonary hypertension ( PH) admitted in Peking Union Medical College Hospital from January 2000 to August 2015 were retrospectively reviewed.Results Total 912 BD patients were hospitalized during the same period, among whom 25 cases were complicated with PH accounting for 3%.There were 15 males (60%) and 10 females (40%), with the mean age of (33 ±12) years (range 19 to 66 years).The median interval from the diagnosis of BD to the onset of PH was 1 year (range 0 to 40 years).The most common cause of PH was heart valve disease (n=10, 40%), followed by pulmonary arterial stenosis or occlusion (n=6, 24%), pulmonary arterial aneurysm with thrombus (n=1, 4%), pulmonary thrombosis (n=1, 4%), cardiomyopathy (n=1, 4%);there were no causes identified in 6 cases (24%).Pulmonary arterial pressures estimated by echocardiography were 40 to 117 mmHg (1 mmHg=0.133 kPa) with a mean pressure of (60 ±22) mmHg.When the PH developed, elevated erythrocyte sedimentation rate ( ESR ) and increased hypersensitive C-reactive protein ( hs-CRP ) were founded in 48% ( 11/23 ) and 82% ( 14/17 ) of patients, respectively.After treated with glucocorticoid (96%,24/25), immunosuppressive agents(92%, 23/25), anticoagulation or thrombolysis(36%,9/25) and specific targeted vasodilator(32%,8/25), the levels of ESR and hs-CRP declined in 91% (10/11) and 71%(10/14) of patients, respectively; and pulmonary arterial systolic pressure declined in 50% of cases (8/16).Among 9 patients followed for 2 to 96 months, 4 died, 1 aggravated, 2 kept stable and 1 improved.Conclusion PH is an uncommon complication in disease.Heart valve disease, pulmonary artery involvement are the major causes of PH.The therapeutic effect and prognosis are poor.

Objective:To explore the efforts of applying mindfulness stress reduction therapy in alleviating stress reactions and burnout among head nurses in the context of the Coronavirus disease 2019 (COVID-19).Methods:This was a quasi experimental study. In June 2021, 109 head nurses with mild or above burnout measured by Maslach Burnout Inventory (MBI) from 12 hospitals in Zhangjiagang City were selected as the research objects to implement mindfulness-based stress reduction therapy for 8 weeks, and the scores of the Five Facet Mindfulness Question-naire (FFMQ), Stress Response Questionnaire (SRQ) and MBI of the head nurses were compared before and after the intervention.Results:The five dimensions of observation, description, perceived action, non-judgment, and non-response and the total scores of the FFMQ were (26.28 ± 1.32), (26.94 ± 1.29), (25.49 ± 0.99), (25.45 ± 1.05), (23.48 ± 1.01), and (127.63 ± 3.78) of the post-intervention, which were higher than the pre-intervention scores of (25.04 ± 1.37), (25.04 ± 1.37), (25.37 ± 1.18), (24.15 ± 1.00), (24.09 ± 0.98), (22.26 ± 1.04), and (120.90 ± 4.06), with statistically significant differences ( t values were -39.21- -15.36, all P<0.01). The three dimensions of emotional, somatic, and behavioral responses and total scores of the post-intervention SRQ were (27.70 ± 6.12), (20.75 ± 4.20), (15.19 ± 3.11), and (65.99 ± 12.43), respectively, lower than the pre-intervention scores of (29.19 ± 6.91), (21.86 ± 5.20), (16.48 ± 4.16), and (70.12 ± 15.97), with statistically significant differences ( t values were 5.70-9.33, all P<0.01); the scores of dimensions of emotional exhaustion and depersonalization in post-intervention MBI were (26.24 ± 4.60) and (5.96 ± 1.25), lower than the pre-intervention scores of (29.66 ± 6.02) and (6.90 ± 1.59). The post-intervention scores of personal fulfillment (32.37 ± 5.02), higher than the pre-intervention scores of (28.60 ± 6.04), all with statistically significant differences ( t=15.36, 13.50, -9.65, all P<0.01). Conclusions:The implementation of mindfulness stress reduction therapy for the head nurses can relieve their work pressure due to the prevention and control of the COVID-19 epidemic, reshape their healthy psychology, and reduce job burnout.

Objective:To explore the relationship between constitutional types of Chinese medicine and Alzheimer's disease (AD) and to construct an early warning model for AD risk.Methods:In the established multimodal interventions to delay dementia and disability in rural China (MIND-China) study, 4 033 elderly subjects aged ≥60 years old were included. The data including demographic, underlying disease and neuropsychological data were collected.The Chinese medicine service record form for the elderly was used to assess constitutional types of Chinese medicine and to apply the NIA-AA diagnostic criteria published by the National Institute on Aging and the Alzheimer's Association in 2011 for the diagnosis of clinically likely AD. Logistic regression analysis and AD risk prediction models were constructed using R statistical software, and the final prediction results were presented using columnar plots.Results:The MIND-China cohort was dominated by the abnormal constitution (69.28%), of which Phlegm-wetness type was the most common (58.05%), followed by Yang-deficiency type (23.85%). The most constitutional type of Chinese medicine among AD patients was Phlegm-wetness type (54.35%), followed by Qi-depression type (38.04%). Multi-factorial logistic regression analysis suggested that increasing age ( β=0.101, P<0.001, OR=1.107, 95% CI=1.069-1.146) and Qi-depression type ( β=0.622, P=0.016, OR=1.862, 95% CI=1.116-3.076) were able to increase the risk of developing AD, while education ( β=-1.047, P<0.001, OR=0.351, 95% CI=0.205-0.584) was able to reduce the risk of developing AD. By using the risk score model to calculate the total risk score for each subject and plotting the receiver operating characteristic curve (ROC), the area under the ROC was 0.769 and the calibration curve showed excellent consistency between prediction and reality. Conclusion:Older adults with Qi-depression type are significantly associated with an increased likelihood of AD.

Objective To explore the relationship between serum nerve growth factor(NGF)and neurofil-amentlight-chain(NF-L)levels with the severity and prognosis of traumatic brain injury(TBI)patients,as well as their predictive value through a prospective research.Methods A total of 132 TBI patients and 132 ordinary trauma patients were collected as the observation group and the control group,respectively.TBI pa-tients in the observation group were divided into three subgroups of mild,moderate,and severe according to Glasgow Coma Scale(GCS)scores,and they were also divided into good and poor subgroups according to Glasgow Outcome Scale(GOS)scores at 6-month follow-up.Enzyme-linked immuno sorbent assay(ELISA)was used to detect serum NGF and NF-L levels in all patients after admission.Through comparison between the observation group and the control group,the relationship between TBI and serum NGF and NF-L levels was clarified.By comparing the differences between different subgroups of the observation group,the relation-ship between serum NGF and NF-L levels and the severity and prognosis of TBI was determined.Through correlation analysis,receiver operating characteristic(ROC)curve and multivariate Logistic regression analy-sis,and the predictive value of serum NGF and NF-L levels for TBI was clarified.Results Compared with the control group,serum NGF and NF-L levels were significantly increased while GCS scores were significantly decreased in the observation group(P＜0.01).In addition,serum NGF and NF-L levels gradually increased a-mong subgroups of mild,moderate,and severe groups in the observation group(P＜0.01).The area under the curve(AUC)of serum NGF and NF-L levels for predicting mild TBI were 0.924(95％CI:0.889-0.986)and 0.991(95％CI:0.982-1.000),and the cut off values were 12.5 ng/mL and 66.5 μg/mL,respectively.AUC of serum NGF and NF-L levels for predicting moderate TBI were 0.776(95％CI:0.686-0.862)and 0.837(95％CI:0.763-0.911),respectively,and AUC for predicting severe TBI was 0.950(95％CI:0.900-0.987)and 0.988(95％CI:0.971-1.000),respectively,with cut off values of 14.5 ng/mL and 157.0 μg/mL,respec-tively.Spearman correlation analysis showed that serum NGF and NF-L levels were negatively correlated with GOS scores of TBI patients(P＜0.01).AUC for predicting the prognosis of TBI patients with serum NGF and NF-L levels were 0.858(95％CI:0.776-0.932)and 0.909(95％CI:0.824-0.978),respectively,with cut off values of 13.5 ng/mL and 71.5 μg/mL,respectively.Logistic regression analysis showed that serum NGF and NF-L levels were risk factors affecting the prognosis of TBI patients.Conclusion Serum NGF and NF-L levels are significantly elevated in TBI patients and could provide reference for early assessment of inju-ry severity in clinical practice.Furthermore,elevated serum NGF and NF-L levels are associated with poor prognosis of TBI patients and are good biological indicators for predicting prognosis.

The CRISPR/Cas9 gene editing technology directs Cas9 protein to recognize, bind and cleave the target site specifically by using artificial single-guide RNA (sgRNA), through non-homologous end joining or homologous end-recombinant repair mechanisms of cells, which can be engineered to knockout or knock-in of genomes. RIG-I is a pattern recognition receptor that recognizes the 5'-triphosphate-containing RNA in the cytoplasm and activates IRF3/7 and NF-κB by interacting with the downstream signaling molecule MAVS, thus initiating the expression of type I interferons and inflammatory factors. Previous studies found that influenza B virus (IBV) can up-regulate the expression of RIG-I. In the present study, to explore whether RIG-I is the major receptor for IBV to active the antiviral innate immune response and its effect on IBV replication, RIG-I gene in 293T cells was knocked out by CRISPR-Cas9 system, and a stable RIG-I knockout 293T (RIG-I(-/-) 293T) cell line was screened by puromycin pressure. The results of Western blotting showed that RIG-I was not expressed in this cell line after IBV or Sendai virus (SeV) infection, indicating that the RIG-I(-/-) 293T cell line was successfully constructed. The transcription levels of interferons, inflammatory factors and interferon-stimulated genes in RIG-I(-/-) 293T cells which were infected by IBV decreased significantly compared with those in wild-type 293T cells. Moreover, the phosphorylation of p65 and IRF3 were not detected in IBV or SeV infected RIG-I(-/-) 293T cells. It is indicated that the expression of cytokines mainly depends on the RIG-I-mediated signaling pathway at the early stage of IBV infection. Furthermore, the multi-step growth curves of IBV in the wild type and RIG-I(-/-) 293T cells showed that RIG-I inhibited the replication of IBV. Collectively, the RIG-I knockout 293T cell line was successfully constructed. We found that RIG-I is the main receptor for IBV to active the antiviral innate immune response and is critical for inhibiting IBV replication, which lays the foundation for further study of IBV infection mechanism.

H9 subtype avian influenza virus causes worldwide epidemic, resulting in enormous economic losses of poultry production. In the present study, an indirect ELISA method was established for more accurate and specific detection. The recombinant protein of the globular head domain of HA of H9 subtype avian influenza virus was used as antigen. Specific blocking buffers and dilution buffers were determined to increase the sensitivity and specificity. The sensitivity of ELISA was higher than that of hemagglutination inhibition (HI) test. The coating antigen is very specific and no cross-reactivity with positive serum against H3N2, H5N2 and H7N9 subtype influenza viruses, Newcastle disease virus, avian infectious bronchitis virus, avian infectious disease virus, and egg drop syndrome virus. Two hundred of clinical sera samples were examined. The results indicate the coincidence rate between ELISA and HI test reached 97%. In addition, there was a positive correlation between OD450 values and the logarithm of HI titer to the base 2 of an individual serum sample (R2=0.981 1).