Objective Optimizing the preparation method to improve the quality of mouse lung cryosections to help enhance the specificity of immunofluorescence staining and obtain more accurate and reliable experimental result.Methods C57BL/6 mouse lung tissue was used to make cryosections via the traditional post-freezing fixation method,pre-freezing fixation method,and a modified perfusion pre-freezing fixation method.A laser-scanning confocal microscope was used to observe lung tissue immunofluorescence staining.Whole areas of mouse lung slices were scanned by fluorescence microscope,and the numbers of intact airways per unit area of lung tissue were calculated.Results In the lung cryosections made via the traditional post-freezing fixation method,the alveoli structure was damaged,the airway wall was seriously disrupted,and there was non-specific staining.Lung cryosections made via the pre-freezing fixation method showed relatively intact alveolar and airway structures but collapsed alveoli and several destroyed airways.In the lung cryosections obtained via the modified perfusion pre-freezing fixation method,the structure and morphology of the alveoli and airways were intact and clear.Additionally,the locations of multiple proteins targeted with immunofluorescence staining were accurate.The number of intact airways(diameter ≥100 μm)per unit area in the lung cryosections obtained via the modified perfusion pre-freezing fixation method was higher than that from cryosections made using the pre-freezing fixation method((0.66±0.15)/mm2 vs(0.33±0.14)/mm2,P＜0.05)and was also significantly higher than that from sections made using the traditional post-freezing fixation method((0.66±0.15)/mm2 vs(0.02±0.04)/mm2,P＜0.01).Conclusions The modified perfusion pre-freezing fixation method for cryosections is conducive to maintaining the integrity of mouse lung tissue morphology and obtaining high-quality multiplex immunofluorescence staining result.

Background::Although existing mycological tests (bronchoalveolar lavage [BAL] galactomannan [GM], serum GM, serum (1,3)-β-D-glucan [BDG], and fungal culture) are widely used for diagnosing invasive pulmonary aspergillosis (IPA) in non-hematological patients with respiratory diseases, their clinical utility in this large population is actually unclear. We aimed to resolve this clinical uncertainty by evaluating the diagnostic accuracy and utility of existing tests and explore the efficacy of novel sputum-based Aspergillus assays. Methods::Existing tests were assessed in a prospective and consecutive cohort of patients with respiratory diseases in West China Hospital between 2016 and 2019 while novel sputum assays (especially sputum GM and Aspergillus-specific lateral-flow device [LFD]) in a case-controlled subcohort. IPA was defined according to the modified European Organization for Research and Treatment of Cancer/Mycoses Study Group criteria. Sensitivity and specificity were computed for each test and receiver operating characteristic (ROC) curve analysis was performed. Results::The entire cohort included 3530 admissions (proven/probable IPA = 66, no IPA = 3464) and the subcohort included 127 admissions (proven/probable IPA = 38, no IPA = 89). Sensitivity of BAL GM (≥1.0 optical density index [ODI]: 86% [24/28]) was substantially higher than that of serum GM (≥0.5 ODI: 38% [39/102]) ( χ2 = 19.83, P < 0.001), serum BDG (≥70 pg/mL: 33% [31/95]) ( χ2 = 24.65, P < 0.001), and fungal culture (33% [84/253]) ( χ2 = 29.38, P < 0.001). Specificity varied between BAL GM (≥1.0 ODI: 94% [377/402]), serum GM (≥0.5 ODI: 95% [2130/2248]), BDG (89% [1878/2106]), and culture (98% [4936/5055]). Sputum GM (≥2.0 ODI) had similar sensitivity (84% [32/38]) (Fisher’s exact P = 1.000) to and slightly lower specificity (87% [77/89]) ( χ2 = 5.52, P = 0.019) than BAL GM (≥1.0 ODI). Area under the ROC curve values were comparable between sputum GM (0.883 [0.812-0.953]) and BAL GM (0.901 [0.824-0.977]) ( P = 0.734). Sputum LFD had similar specificity (91% [81/89]) ( χ2 = 0.89, P = 0.345) to and lower sensitivity (63% [24/38]) ( χ2 = 4.14, P = 0.042) than BAL GM (≥1.0 ODI), but significantly higher sensitivity than serum GM (≥0.5 ODI) ( χ2 = 6.95, P = 0.008), BDG ( χ2 = 10.43, P = 0.001), and fungal culture ( χ2 = 12.70, P < 0.001). Conclusions::Serum GM, serum BDG, and fungal culture lack sufficient sensitivity for diagnosing IPA in respiratory patients. Sputum GM and LFD assays hold promise as rapid, sensitive, and non-invasive alternatives to the BAL GM test.

The Laboratory of Pulmonary Diseases at West China Hospital in Sichuan University starts with strengthening the laboratory safety education and research integrity, develops a special individualized experimental skill training, improves communication between project team and laboratory, and promotes the cooperation between the laboratory and university academic community. Combined with the new model of project hierarchical management, the "Innovation and Enterpreneurship Training Programs for Undergraduates" are dynamically evaluated and effectively supervised. With the implementation of the new management mode, the number of projects successfully obtained has increased by 16.7%, the number of published academic papers has increased by 6.5 times, and the quality of published papers has been also significantly improved.

Objective:To summarize the status and trends of papers published by Chinese scholars on the combination teaching model in the area of medical education in China.Methods:Literature retrieval was performed on CNKI, WanFang, and VIP database, and studies regarding to combination teaching model in medical education were selected and analyzed. BICOMS-2 analysis software was used to extract and sort out keywords, provinces, authors and their organizations and generate co-occurrence matrix and/or word matrix. NetDraw and gCLUTO software were used to draw the network relationship diagram and cluster analysis respectively.Results:Totally 99 articles were included this study, and the number of articles published each year showed an increasing trend. Authors from 15 provinces contributed to these articles, with uneven development and limited cooperation across regions. The authors were divided into 4 main groups, but the communication among these groups was not enough. And there were 3 main research topics.Conclusion:The number of articles on combination teaching model in medical education in China is increasing, covering many authors, teams, institutions and regions. However, the communication and cooperation among these authors and institutions needs to be strengthened; and the research topics should be expanded.

Objective:To investigate the cognition of the diagnosis and management of bronchiectasis among respiratory specialists.Methods:Between May and June 2020, a questionnaire survey based on expert consensus and guidelines was conducted among respiratory specialists from 50 hospitals from Sichuan and Yunnan provinces. Total 691 questionnaires were distributed and 641 were received. 601 valid questionnaires were chosen for further analysis with an effective recovery rate of 87.0%. The respondents were required to finish the e-questionnaires independently in terms of cognition of diagnosing, severity assessment, treatments and Chinese medicines of bronchiectasis. The responses were collected online and the cognitive levels were evaluated by calculating the correct rates of corresponding questions.Results:540 (89.9%) of the respondents agreed that high-resolution computed tomography (HRCT) was the gold standard for diagnosing of bronchiectasis, but 318 (52.9%) had an incomplete understanding of common radiographic manifestations of bronchiectasis, and different cognitive degrees of common radiographic manifestations of bronchiectasis existed among respondents with different qualifications or working in different levels of hospitals ( P<0.05). Only 118 (19.6%) of the respondents were familiar with severity assessments of bronchiectasis, but 65 (55.1%) of the 118 respondents said they won′t apply these severity assessments to patient during their clinical works. For the treatment of patients with stable bronchiectasis, airway clearance techniques were most recommended by specialists surveyed [410 (68.2%)], among which, postural drainage was the most known method [559 (93.0%)]. For patients undergoing an acute exacerbation, most respondents recommended antibiotics [600 (99.8%)] as the primary treatment, and examinations such as sputum culture [544 (90.5%)], inflammatory markers [523 (87.0%)] should be performed as well. 504 (83.9%) thought that pseudomonas aeruginosa was the most common conditioned pathogen for bronchiectasis. For patients with frequent exacerbations (≥3 per year), 385 (64.1%) of the respondents supported the therapeutic effect of long-term antibiotics, however, among the 385 respondents supporting long-term antibiotics, only [113 (29.4%)] were willing to recommend long-term antibiotic treatment actively during their clinical practice. Besides, 304 (50.6%) respondents held a positive attitude to the clinical effect of traditional Chinese medicine therapies. Among 304 respondents holds the point of supporting, only 86 (28.3%) were willing to recommend traditional Chinese medicine to patients actively, differences about the attitude and clinical behaviors were found between respondents working in different levels of hospitals ( P<0.05). Conclusions:Respiratory specialists′ cognition on diagnosis and treatments of bronchiectasis remains inadequate, and cognitive levels differ among respondents working in different levels of hospitals. There is a gap between respondents′ cognition and clinical practice. Further education and trainings are necessary for improving respiratory specialists′ knowledge for timely diagnosis and standard treatment of bronchiectasis.

Sensory processing is strongly modulated by different brain and behavioral states, and this is based on the top-down modulation. In the olfactory system, local neural circuits in the olfactory bulb (OB) are innervated by centrifugal afferents in order to regulate the processing of olfactory information in the OB under different behavioral states. The purpose of the present study was to explore the organization of neural networks in olfactory-related cortices and modulatory nuclei that give rise to direct and indirect innervations to the glomerular layer (GL) of the OB at the whole-brain scale. Injection of different recombinant attenuated neurotropic viruses into the GL showed that it received direct inputs from each layer in the OB, centrifugal inputs from the ipsilateralanterior olfactory nucleus (AON), anterior piriform cortex (Pir), and horizontal limb of diagonal band of Broca (HDB), and various indirect inputs from bilateral cortical neurons in the AON, Pir, amygdala, entorhinal cortex, hippocampus, HDB, dorsal raphe, median raphe and locus coeruleus. These results provide a circuitry basis that will help further understand the mechanism by which olfactory information-processing in the OB is regulated.

Objective To compare clinical characteristics between acute exacerbated chronic obstructive pulmonary disease (AECOPD) patients with chronic bronchitis phenotype and emphysema phenotype.Methods A total of 73 AECOPD patients with chronic bronchitis phenotype and 82 AECOPD patients with emphysema phenotype in Tibet Chengdu Branch Hospital,West China Hospital,Sichuan University from January 2014 to November 2016 were selected.All patients had completed lung function tests,basic information collection,modified medical research council dyspnea scale (MMRC),and COPD assessment test (CAT).The serum samples were collected to detect C-reactive protein (CRP).The clinical characteristics,lung function,complications and systemic inflammation between the two groups were compared.Results There was no difference in body mass index (BMI),CAT score,serum CRP level and length of hospital stay between the AECOPD patients with chronic bronchitis phenotype and emphysema phenotype (P＞0.05).Compared with AECOPD patients with chronic bronchitis phenotype,the AECOPD patients with emphysema phenotype have longer smoking history,higher MMRC score,and more severe lung function impairment (P＜0.05).Conclusion AECOPD patients with chronic bronchitis phenotype and emphysema phenotype have different clinical characteristics,those with emphysema phenotype have more severe dyspnea and lung function impairment.

OBJECTIVE: To study the pathologic characteristics of bone marrow for CD5 positive small B cell lymphoma (SBL).
 METHODS: The pathologic profiles of 92 patients with CD5 positive SBL were retrospectively analyzed. The morphologic and immunophenotypic features were analyzed by flow cytometry and immunohistochemistry. IgH/CCND1 was examined by fluorescence in situ hybridization (FISH).
 RESULTS: A total of 92 patients with CD5 positive SBL were enrolled in this study, including 56 (60.9%) chronic lymphocytic leukemia /small lymphocytic lymphoma (CLL/SLL), 23 (25.0%) mantle cell lymphoma (MCL) and 13 other SBL (14.1%). Among the 13 other cases, 5, 4 and 4 cases were follicular lymphoma (FL), lymphoplasmacytic lymphoma (LPL) and splenic marginal zone lymphoma (SMZL), respectively. The frequency of patterns for bone marrow infiltration was as follow: diffuse pattern (19/92), mixed pattern (15/92), nodular pattern (9/92), interstitial pattern (8/92), and intrasinusodial pattern (2/92). All patients expressed CD19, CD20 and CD5. According to the immunophenotypic score system, all the CLL patients had 4-5 scores, while SMCL and other SBL patients had less than 3 scores. For the other SBL patients, 5 FL expressed CD10, while 3 FL, 1 LPL and 3 SMZL expressed CD23. There was a significant difference in the expression of CD23, sIgM, FMC7, CD11C and CD22 between the CLL and MCL groups (P<0.01). All 23 MCL patients expressed cyclin D1 and showed IgH/CCND1 gene translocation by FISH detection.
 CONCLUSION CD5 positive SBL includes a variety of types of lymphoma. Patterns of bone marrow for CD5 positive SBL are diversity. Immunophenotypic analysis by flow cytometry is essential in the diagnosis and differential diagnosis of CD5 positive SBL, especially for CLL.
Bone Marrow ; pathology ; CD5 Antigens ; metabolism ; Diagnosis, Differential ; Flow Cytometry ; Humans ; Immunohistochemistry ; In Situ Hybridization, Fluorescence ; Leukemia, Lymphocytic, Chronic, B-Cell ; diagnosis ; Lymphoma, B-Cell ; diagnosis ; Lymphoma, Follicular ; diagnosis ; Lymphoma, Mantle-Cell ; diagnosis ; Oncogene Proteins, Fusion ; metabolism ; Retrospective Studies ; Splenic Neoplasms ; diagnosis

Bronchial Diseases ; diagnosis ; etiology ; Child, Preschool ; Humans ; Male ; Pulmonary Artery ; pathology

BACKGROUNDCigarette smoke induced airway inflammation plays a role in pathogenesis of airway inflammation. Resolvin-D1 derived from omega-3 polyunsaturated fatty acids is an endogenous anti-inflammatory and proresolving lipid mediator. Resolvin-D1 ameliorated inflammatory responses in lung injury, asthma, peritonitis and atherosclerosis. We investigated whether resolvin-D1 suppressed the productions of chemokines and oxidative stress induced by cigarette smoke extract (CSE) in vitro and its possible mechanism.

METHODSWe examined the proinflammatory chemokine interleukin-8 and hydrogen peroxide (H2O2) productions induced by CSE in 16 human bronchial epithelial (16HBE) cells after resolvin-D1 treatment and their mechanisms. 16HBE cells were treated with resolvin-D1 at up to 10 nmol/L, for 30 minutes before CSE up to 16% (v/v) exposure. Release of interlukin-8 proteins was assessed by enzyme linked immunosort assay (ELISA) and its mRNA level by RT-PCR. We evaluated extracellular H2O2 expression in the supernatant. Phosphorylation of NF-κB/p65 and degradation of I-κB in 16HBE cells were determined by Western blotting analysis and NF-κB DNA binding activity by electrophoretic mobility shift assay (EMSA).

RESULTS16HBE cells treated with 8% CSE showed significantly higher interlukin-8 production. Resolvin-D1 pretreatment inhibited CSE induced interlukin-8 production (mRNA and protein) in a dose and time dependent manner. Extracellular H2O2 level decreased after resolvin-D1 treatment. Resolvin-D1 attenuated CSE triggered I-κB degradation and NF-κB/p65 activation dose dependently and inhibited NF-κB DNA binding activity.

CONCLUSIONResolvin-D1 inhibits CSE induced interlukin-8 and H2O2 production in 16HBE cells by modulating NF-κB activation and has therapeutic potential for pulmonary inflammation.

Blotting, Western ; Cell Line ; Cell Survival ; drug effects ; Docosahexaenoic Acids ; pharmacology ; Electrophoretic Mobility Shift Assay ; Enzyme-Linked Immunosorbent Assay ; Humans ; Hydrogen Peroxide ; metabolism ; Interleukin-8 ; metabolism ; NF-kappa B ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Smoking ; adverse effects