Alkaloids from Stems of Ervatamia yunnanensis （Ⅱ）

Six indole alkaloids were isolated from the stems of Ervatamia yunnanensis. Among them, 10-hydroxy-19, 20-dihydroisositsirikine （I） is new. The other five are known alkaloids, namely： matrine （Ⅱ）, 19, 20-dihydroisositsirikine （Ⅲ）, 19-s-voacangarine （Ⅳ）, 1 l＇-methoxyl-19s- heyneanine （Ⅴ）, conodurine （Ⅵ）. The structural elucidation of the alkaloids was based on spectral means.

Alkaloids from stems of Ervatamia yunnanensis

Six indole alkaloids were isolated from the stems of Ervatamia yunnanensis. Among them, yunnanensine (I) is new. The other five are known alkaloids, namely: 19,20-E-vallesamine (II), 19s-heyneanine (III), ibogaine (IV), ibogamine (V), coronaridine (VI). The structural elucidation of the alkaloids was based on spectral means.

A SARS-CoV-2 neutralizing antibody discovery by single cell sequencing and molecular modeling

Although vaccines have been developed,mutations of SARS-CoV-2,especially the dominant B.1.617.2(delta)and B.1.529(omicron)strains with more than 30 mutations on their spike protein,have caused a significant decline in prophylaxis,calling for the need for drug improvement.Antibodies are drugs preferentially used in infectious diseases and are easy to get from immunized organisms.The current study combined molecular modeling and single memory B cell sequencing to assess candidate sequences before experiments,providing a strategy for the fabrication of SARS-CoV-2 neutralizing antibodies.A total of 128 sequences were obtained after sequencing 196 memory B cells,and 42 sequences were left after merging extremely similar ones and discarding incomplete ones,followed by homology modeling of the antibody variable region.Thirteen candidate sequences were expressed,of which three were tested positive for receptor binding domain recognition but only one was confirmed as having broad neutralization against several SARS-CoV-2 variants.The current study successfully obtained a SARS-CoV-2 antibody with broad neutralizing abilities and provided a strategy for antibody development in emerging infectious diseases using single memory B cell BCR sequencing and computer assistance in antibody fabrication.

The prognosis and short-term efficacy of corticosteroid therapy for COVID-19 patients

Dear Editor:Corticosteroids are widely used in the treatment of critically ill patients for active anti-inflammatory and immunosuppressive effects.However,the therapeutic effects of corticosteroid treatment are still disputable.Some studies have shown that corticosteroids can delay coronavirus RNA clearance[1],increase mortality[2]and cause adverse events[3].Conversely,other studies have confirmed the clinical value of corticosteroids in shortening hospital stays and decreasing the need for mechanical ventilation[4].

Gene transfer of a β_2-adrenergic receptor kinase inhibitor up-regulates the level of β_2-adrenergic receptor and cAMP in the asthmatic murine lung

Objective： To investigate the effects of gene transfer of a β-adrenergic receptor（β-AR） kinase inhibitor（β ARIct） on pulmonary β2-adrenergic receptor and cAMP following β2-AR agonist treatment in asthmatic mice, and to analyze the relationship between the routes of gene delivery and the changes of β2AR and cAMP. Methods： BALB/c mice were sensitized and challenged by ovalbumin to establish the asthmatic model treated with βAR agonist （salbutamol injected intramuscularly）. The plasmid with the expression of βARKct was constructed and βARKct gene transfer was performed through intravenous injection or intratracheal instillation in asthmatic mice. The gene expression was measured with Western blot analysis, and the changes of pulmonary β-AR and cAMP evaluated by Radioimmunoassay. Results： The expression of tranfered βARKct gene was detectable in lungs and it was expressed more in the lungs of the mice receiving intratracheally plasmid than those receiving intravenously. The levels of βAR and cAMP were upregulated after using plasmid-βARKct to the asthmatic mice treated with βAR agonist. Conclusion： Our results indicated that there were down-regulation of βAR and cAMP in asthmatic mice treated with βAR agonist. Gene transfer of βARKct could inhibit the extent of the down-regulation of βAR and cAMP. The route of gene delivery could also affect the degree of up-regulation of βAR and cAMP. Gene transfer βARKct may provide a novel approach to the therapeutic strategy for asthma.

Endobronchial Ultrasound Elastography for Diagnosing Mediastinal and Hilar Lymph Nodes

Background： Pathophysiological processes, such as malignancy, can lead to the formation of stiffer tissue in lung cancers. Endobronchial ultrasound rEBUS） elastography is a novel technique for measuring tissue stiffness during EBUS-guided transbronchial needle aspiration （EBUS-TBNA）. The current study was conducted to investigate the diagnostic value of EBUS elastography tbr mediastinal and hilar lymph node metastasis in lung cancers. Methods： From January 2014 to January 2015, 40 patients suspected of lung cancer were enrolled, and a total of 68 lymph nodes were evaluated by EBUS-TBNA. EBUS-guided elastography of lymph nodes was perfornaed prior to EBUS-TBNA. Standard EBUS characteristics were also described. Pathological determination of malignant or benign lymph nodes was used as the gold standard for this study. If EBUS-TBNA did not result in a tbrmal pathological diagnosis of malignancy, patients were referred for a surgical procedure. Comparisons of elastography and standard EBUS characteristics were made between benign and malignant lymph nodes. Results： Elastography grading scores and strain ratios showed significant differences between benign and malignant lymph nodes （P = 0.000）. The elastography strain ratio was more sensitive and specific for determining malignant lymph nodes than elastography grading score or standard EBUS criteria. The receiver operating characteristic curve for the elastography strain ratio showed an area under the curve of 0.933. The best cut-off point of the strain ratio for differentiating malignant from benign lymph nodes was 32.07. The elastography strain ratio had a sensitivity of 88.1%, the specificity of 80.8%, positive predictive value of 88.1%, and negative predictive value of 80.8% for distinguishing malignant from benign nodes. The overall accuracy of elastography strain ratio was 85.3%. The strain ratio of malignant and benign lymph nodes positively correlated with the elastography grading score （r = 0.561, P = 0.000）. Conclusions： EBUS elastography can be effectively used to predict mediastinal and hilar lymph node metastases in lung cancer. This noninvasive technique may thus complement standard EBUS and help guide EBUS-TBNA procedures.

Eligibility of C-BIOPRED severe asthma cohort for type-2 biologic therapies

To the Editor:Patients with severe persistent asthma experience greater morbidity with more impairment in quality of life despite higher use of health care resources and being treated with existing asthma treatments such as inhaled corticosteroids and b-agonists,and sometimes oral corticosteroid(OCS)therapy.Type-2(T2)high asthma has been identified as a phenotype that responds to targeted T2 biologic therapies such as anti-IgE,anti-interleukin(IL)5,or anti-IL5Ra and anti-IL4Ra monoclonal antibodies,which are currently available in Europe and North America,and are currently introduced in the rest of the world.[1]

Downregulation of miR-4772-3p promotes enhanced regulatory T cell capacity in malignant pleural effusion by elevating Helios levels

Background:Malignant pleural effusion(MPE)is a complicated condition of patients with advanced tumors.Further dissecting the microenvironment of infiltrated immune cells and malignant cells are warranted to understand the immune-evasion mechanisms of tumor development and progression.Methods:The possible involvement of microRNAs(miRNAs)in malignant pleural fluid was investigated using small RNA sequencing.Regulatory T cell(Treg)markers(CD4,CD25,forkhead box P3),and Helios(also known as IKAROS Family Zinc Finger 2[IKZF2])were detected using flow cytometry.The expression levels of IKZF2 and miR-4772-3p were measured using quantitative real-time reverse transcription polymerase chain reaction.The interaction between miR-4772-3p and Helios was determined using dual-luciferase reporter assays.The effects of miR-4772-3p on Helios expression were evaluated using an in vitro system.Correlation assays between miR-4772-3p and functional molecules of Tregs were performed.Results:Compared with non-malignant controls,patients with non-small cell lung cancer had an increased Tregs frequency with Helios expression in the MPE and peripheral blood mononuclear cells.The verified downregulation of miR-4772-3p was inversely related to the Helios*Tregs frequency and Helios expression in the MPE.Overexpression of miR-4772-3p could inhib让Helios expression in in vitro experiments.However,ectopic expression of Helios in induced Tregs reversed the effects induced by miR-4772-3p overexpression.Additionally,miR-4772-3p could regulate Helios expression by directly targeting IKZF2 mRNA.Conclusion:Downregulation of miR-4772-3p,by targeting Helios,contributes to enhanced Tregs activities in the MPE microenvironment.