[System analysis of the ecological distribution of bacteriophages in hospital wastewater].

Phage therapy is one of the most important tools for the treatment of infections with multi-drug resistant bacteria. Such phages are usually isolated from hospital effluents, however, no systematic study on the distribution of phages in hospital effluents has been conducted so far. The aim of this study was to isolate the corresponding phages of common pathogenic bacteria isolated in the clinic as hosts, so as to assess the ecological distribution of phages in hospital wastewater and to provide a reference for the isolation and application of phages of drug-resistant bacteria in the clinic. A cross-sectional study design was used in this study. The 125 pathogenic bacteria (belonging to 16 different strains) isolated from the clinical microbiology laboratory of Qilu Hospital of Shandong University from May to June 2023 were selected as the target strains, and the phages corresponding to these strains were isolated and purified from the hospital wastewater by using the double-layer plate sandwich method. At the same time, the distribution of pathogenic bacteria in the same batch of wastewater was analyzed with the help of mNGS sequencing technology, so as to preliminarily investigate the abundance correspondence between pathogenic bacteria and phages in wastewater. The results showed that a total of 56 phage strains were isolated from 125 clinical pathogens as hosts, corresponding to six pathogens, including Acinetobacter baumannii, Klebsiella pneumoniae, Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, and Stenotrophomonas maltophilia. All six pathogenic bacteria contained strains with different degrees of drug resistance, with a higher percentage of multi-drug resistant strains in A. baumannii, Escherichia coli and P. aeruginosa. The phage acquisition rates of these six pathogens were, in descending order, Escherichia coli (80%), Stenotrophomonas maltophilia (75%), Pseudomonas aeruginosa (70%), Klebsiella pneumoniae (66.67%), Acinetobacter baumannii (36.36%) and Staphylococcus aureus (12.5%). Preliminary mNGS sequencing results showed that the pathogenic bacteria with higher abundance in the batch of effluent were Enterococcus faecalis, Klebsiella pneumoniae, Escherichia coli, Enterococcus faecalis, Acinetobacter baumannii, Klebsiella aerogenes, Klebsiella michiganensis and Pseudomonas aeruginosa. In conclusion, phages of most common clinical Gram-negative pathogens were isolated from hospital wastewater with high isolation rates; however, phages of Gram-positive pathogens were isolated at lower rates, and only phages corresponding to Staphylococcus aureus were isolated in this study. The corresponding mNGS sequencing results showed that the distribution of Gram-negative pathogens in sewage may had a positive correlation with the ecological distribution of phages.

[Clinical features and reasons for missed diagnosis of allergic bronchopulmonary aspergillosis].

Objective: To summarize the clinical features of allergic bronchopulmonary aspergillosis (ABPA) and analyze the common causes of missed diagnosis. Methods: The clinical data of patients with ABPA who were admitted into Qilu Hospital of Shandong University from October 2014 to November 2017 were retrospectively analyzed, including baseline data, eosinophil count in peripheral blood, serum total IgE, A. fumigates-specific antibody (sIgE, sIgG and sIgM), pulmonary function tests and chest CT, etc. Then the correlations between serum total IgE, sIgE and forced expiratory volume in one second (FEV1) as percentage of predicted value (FEV1%pred) was conducted, as well as that between serum total IgE, sIgE and FEV1/forced vital capacity (FEV1/FVC). The clinical features of the patients were summarized and the causes of missed diagnosis were analyzed. Results: There were 46 patients with ABPA (21 males and 25 females, 48.3±13.2 years old) in total. Only 2 cases (4.3%) were diagnosed after the first symptom onset, and 44 cases (95.7%) were treated repeatedly for many years before being diagnosed. The misdiagnoses were asthma (26 cases, 56.5%), bronchiectasis (13 cases, 28.3%), pneumonia (4 cases, 8.7%), pulmonary tuberculosis (3 cases, 6.5%), and lung cancer (4 cases, 8.7%). Common symptoms of ABPA included cough, expectoration, wheezing, chest tightness, fever, weight loss, chest pain, etc. Peripheral blood eosinophil count increased in 37 patients (80.4%). Serum total IgE in 37 patients (80.4%) were higher than 1 000 U/ml, and 9 cases (19.6%) were less than 1 000 U/ml. The positive rate of sIgE, sIgG and sIgM was 100.0%, 89.1% and 54.3%, respectively. All patients underwent pulmonary function tests and 35 cases (76.1%) had obstructive ventilation dysfunction, 5 cases (10.9%) with mixed ventilation dysfunction, 5 cases (10.9%) were normal and 1 case (2.2%) with restrictive ventilation dysfunction. All patients underwent Chest CT examination, 28 cases (60.9%) showed bronchiectasis, 8 cases (17.4%) manifested mucus plugs (among them, 4 cases with high-attenuation mucus) and 10 cases (21.7%) had other atypical imaging. Serum total IgE and sIgE had no correlations with FEV1%pred and FEV1/FVC. Conclusions: ABPA is characterized by recurrent episodes of wheezing, fleeting pulmonary opacities and bronchiectasis. The main reasons of misdiagnosis in ABPA patients were: atypical symptoms, variety of predisposing diseases, early atypical imaging changes, limitations and misunderstandings of screening indicators, interference with tumor markers, and the presence of pulmonary aspergillus overlap syndrome. The severity of asthma attacks is not related to the degree of fungal sensitization.

Screening miRNAs related to different subtypes of breast cancer with miRNAs microarray.

AIM: The aim of this study was to screen miRNAs related to different subtypes of breast cancer and their target genes to identify new markers of tumor subtype. MATERIALS AND METHODS: The miRNA expression profiles of breast cancer GSE38867 including 7 ductal carcinoma in situ breast (DCIS) cancer samples, 7 invasive breast cancer samples, 7 metastatic breast cancer samples, and 7 normal breast samples) were downloaded from Gene Expression Omnibus (GEO) database. Limma package in R software was applied to identify specific differentially expressed miRNAs of different subtypes of breast cancer. MicroRNA.org database source was used to predict the target genes of the identified differentially expressed miRNAs. We integrated the target genes and their interacted genes (predicted by STRING) into DAVID to perform the GO function and KEGG pathway analyses. RESULTS: Compared to the normal control, a total of 21, 47, and 107 differentially expressed miRNAs were screened in DCIS, invasive and metastatic breast cancer, respectively. Specific differentially expressed miRNAs of the three subtypes were identified, including hsa-miR-99a and hsa-miR-151-3p for DCIS breast cancer, hsa-miR-145 and hsa-miR-210 for invasive breast cancer, and has-miR-205 and has-miR-361-5p metastatic breast cancer. Furthermore, 220, 43, 446, 307, 587 and 328 interaction pairs of the specific miRNA targets were predicted. Multiple GO functions and KEGG pathways were enriched with the miRNA targets and their interacted genes. CONCLUSIONS: We screened the most representative miRNAs of the three different subtypes of breast cancer, which may act as the putative markers in the diagnosis of different subtypes of breast cancer.