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Objective:To establish the method for detecting lower respiratory infections (LRIs) bacterialpathogens using nanopore sequencing, and evaluate the feasibility of this method.Methods:Bronchoalveolar lavage fluid (BALF) samples from 33 patients with LRIs who visited the Department of Respiratory and Critical Care Medicine of Beijing Hospital from July 2019 to September 2020 were collected.Nanopore 16S amplicon sequencing were performed on these samples. In order to evaluate the clinical value of the nanopore sequencing, χ 2 test was used to analyze the pathogen differences between the detection rate and pathogen types results found with using the nanopore 16S sequencing and the results found with bacterial culture. Results:The process and method of nanopore sequencing used in the detection of the LRIs pathogens were established. The pathogen detection rate of the 16S sequencing was higher than that of the traditional bacterial culture (75.8% [25/33], 45.5% [15/33], χ2=5.140, P<0.05). From the 25 positive samples found with nanopore 16S sequencing, 16 pathogens were detected, including Haemophilus parainfluenzae, Haemophilus influenzae, Streptococcus pneumoniae, Streptomonas maltophilia, Acinetobacter baumannii, and Acinetobacter junii, Staphylococcus aureus, Klebsiella pneumoniae, Enterococcus faecalis, Enterococcus gallinarum, Corynebacterium striatum, Mycobacterium paraintracellulare, Serratia marcescens, Achromobacter insuavis, Citrobacter murliniae and Mycoplasma pneumoniae. More than 6 pathogens were tested in clinical culture, including Haemophilus parainfluenzae, Acinetobacter baumannii, Pseudomonas aeruginosa, Staphylococcus aureus, Klebsiella pneumoniae and Streptomonas maltophilia (χ2=7.949, P<0.05). 16S sequencing aligned to species level sequences accounted for 80.0 (60.0, 86.0)% of the genus level. The results obtained by using16S sequencing and bacterial culture were consistent in 11 (33.3%) samples. Conclusions:Nanopore 16S amplicon sequencing can quickly identify pathogenic bacteria from BALF in LRIs patients. Nanopore 16S amplicon sequencing has a high detection rate, it can detect more pathogens than traditional bacterial culture, and it can also identify most bacteria to the species level. This technology is a very promising platform with broad application prospects.
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Objective:To investigate the changes in clinical signs and laboratory testing results and their risk-tiring diagnostic effectiveness in elderly patients with pulmonary embolization (PE) with different risk levels.Methods:A retrospective analysis was conducted on the clinical data of elderly hospitalized PE patients in Beijing Hospital and other coordinated hospital from 2012 to 2020.Differences in 43 clinical signs and detection indicators between patients with four different risk levels were compared.The univariate and multivariate regression models were used to analyze differences between high-risk and non-high-risk PE and between intermediate-risk and low-risk PE with ROC analysis.Results:In the multi-group comparison, there are 33 clinical tests having significant differences between four risk groups, 29 clinical tests having significant differences between three risk groups(high, intermediate and low groups), and 21 clinical tests having significant differences between two groups(high and non-high groups). In the ROC analysis of risk stratification in high-risk and non-high-risk groups, it was found that the range of area under the curves(AUC)of 14 significantly changed clinical tests were 0.611 to 0.802 in the univariate regression analysis.The AUC of the model of systolic blood pressure(SBP)combined with white blood cell count(WBC)and aspartate aminotransferase(AST)was 0.8593(95% CI: 0.795-0.924)in the multivariate regression analysis.While in the ROC analysis between intermediate-risk and low-risk, the range of AUC of 12 significantly changed clinical tests were 0.592 to 0.835 in the univariate regression analysis.The B-type natriuretic peptide(BNP)and N-terminal B-type natriuretic peptide(NT-proBNP)can assist the risk stratification in intermediate-risk and low-risk PE groups.No efficient combined diagnosis model was found. Conclusions:The basic vital signs and multiple clinical laboratory tests were significantly different among four risk levels of elderly PE patients, such as blood gas analysis, coagulative function, liver and kidney function and myocardial markers.The combination of SBP, WBC, and AST can effectively assist the risk stratification in high-risk and non-high-risk PE groups.
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To investigate whether the laboratory specimens preserved in Beijing Hospital Biobank during a specific period had been contaminated by SARS-Cov-2 through a cross-sectional study, and to establish a retrospective biobank safety screening system. Laboratory specimens were collected from the Department of Respiratory and Critical Care Medicine and the Fever Clinic of Beijing Hospital from November 1, 2019 to January 22, 2020, nucleic acid and serological antibody testing were performed for SARS-CoV-2 in these specimens (including 79 serum, 20 urine, 42 feces and 21 bronchoalveolar lavage fluid specimens). The safety of the stored samples during this period was defined by negative and positive results. Both the nucleic acid test and serological antibody test showed negative for SARS-CoV-2, indicating that these specimens were safely stored in the biobank. High-risk specimens collected in our hospital during the early stage of the COVID-19 outbreak are free of SARS-CoV-2, and a safety screening strategy for the clinical biobank is established to ensure the biosafety of these samples.
Subject(s)
Humans , Biological Specimen Banks , COVID-19 , Cross-Sectional Studies , Hospitals , Retrospective Studies , SARS-CoV-2ABSTRACT
Objective To explore the expression of b-cell lymphoma/leukemia-2 (bcl-2) and breast cancer drug-resistant protein (BCRP) in diffuse large B cell lymphoma (DLBCL) and their correlation. Methods Using flow cytometry (FCM) and semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) assay the expressions of BCRP gene was analysed in 40 DLBCL samples. bcl-2 protein were measured by immunohistochemistry on paraffin-embedded slices in 40 cases of DLBCL. Results The positive expression rate of bcl-2 and BCRP protein were 60.0 % (24/40) and 37.5 % (15/40) in 40 patients with DLBCL respectively. There was significant differences between positive group and negative groups (x2 = 5.7618,P 0.05). Conclusion BCRP may play an important role in DLBCL primary much medicine drug-resistant,thus it may be a useful prediction of chemotherapeutic treatment and risk of relapse. The level bcl-2 expression was closely related with the grade of malignancy and the prognosis of DLBCL. There was no significant correlation between bcl-2 and BCRP gene. The detection of bcl-2 and BCRP gene played an important role in evaluating DLBCL outcome.
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Objective To study clinical characteristics and prognostic factors in systemic anaplastic large cell lymphoma (S-ALCL). Methods Clinical date of 30 systemic ALCL patients from Department Hemotology of Shanxi Cancer Hospital from December 1998 to September 2009 were reviewed. Results The median age of the 30 patients was 36 years old. The male/female ratio was 1.5:1.18 (60.0 %) 18/30 of the patients had B symptoms, 22 (73.3 %) had Ann Arbor stage Ⅲ-Ⅳ, and 18 (60.0 %) had extranodal disease at diagnosis. 14 (46.7 %) had elevated lactate dehydrogenase (LDH) level. The positive rate of ALK was 18 (60.0 %). Compared with ALK negative patients, ALK-positive patients were younger (u = 3.92, P=0.001).ALK expression and LDH level were important prognostic factors by Long-rank test. Conclusion S-ALCL usually occur in younger patients, with good response to chemotherapy and good prognosis. ALK negative and elevated LDH level are correlated to unfavorable prognosis. Combination chemotherapy is the major choice to these patients. High-dose chemotherapy maybe benefit to the patients who have adverse prognostic factors.