Identification and analysis of a clinically isolated strain of Halomonas based on whole-genome sequencing and comparative genomics.

OBJECTIVE: The aim of this study was to identify the species of a Halomonas strain isolated from a neonatal blood sample and to understand the potential pathogenicity and characteristic genes of the strain. METHODS: The genomic DNA of strain 18071143 (identified as Halomonas by matrix-assisted laser desorption-ionization time of flight-mass spectrometry and the 16S ribosomal RNA (rRNA) gene sequence) was sequenced using Nanopore PromethION platforms. The average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) were calculated using the complete genome sequences of the strain. Comparative genomic analyses were performed on strain 18071143 and 3 strains of Halomonas (Halomonas stevensii S18214, Halomonas hamiltonii KCTC 22154, and Halomonas johnsoniae KCTC 22157) that were associated with human infections and had high genomic similarity to strain 18071143. RESULTS: Phylogenetic, ANI, and dDDH similarity analyses based on genome sequence indicated that strain 18071143 belonged to the species H stevensii. Similarities exist between strain 18071143 and the other 3 Halomonas strains in terms of gene structure and protein function. Nonetheless, strain 18071143 has greater potential for DNA replication, recombination, repair, and horizontal transfer. CONCLUSION: Whole-genome sequencing holds great promise for accurate strain identification in clinical microbiology. In addition, the results of this study provide data for understanding Halomonas from the perspective of pathogenic bacteria.

Impact of COVID-19 pandemic on influenza virus prevalence in children in Sichuan, China.

We performed a retrospective analysis of influenza A and B virus antigen detection data in children in Sichuan Province from January 2019 to December 2021, with the goal of studying the impact of the COVID-19 pandemic on influenza circulation in children in Sichuan, China. During the pandemic, both the number of specimens and the positive rates of the influenza virus fell dramatically. The positivity for influenza A virus decreased from 22.5% in 2019 to 9.9% in 2020 to 0.2% in 2021 (p < 0.001). The lowest and highest positive rates for the influenza B virus occurred in 2020 and 2021, respectively, with a statistically significant 3-year comparison (p < 0.001). During the pandemic, the annual positivity remained higher in school-age than in preschoolers, while there was no difference in the annual positivity between the two gender groups, both consistent with the prepandemic results. During the pandemic, the seasonality of influenza A and B was different from that before the pandemic. In 2019, the epidemic season for influenza A was autumn and winter, while the epidemic season for influenza B was winter and spring. Seasonal changes in influenza A were insignificant after the pandemic, and influenza B became predominant in 2021, with a high prevalence in the autumn. Although influenza activity decreased during the COVID-19 pandemic, one should be on the lookout for a possible rebound in influenza circulation in the future.

All-trans retinoic acid reduces cancer stem cell-like cell-mediated resistance to gefitinib in NSCLC adenocarcinoma cells.

BACKGROUND: The enrichment of cancer stem cell-like cells (CSCs) has been considered to be responsible for tumor progression after an initial response to epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (EGFR-TKIs) in patients with non-small cell lung adenocarcinoma (NSCLC/ADC). CSCs with ALDH1A1bright /CD44high expression contribute to the TKIs resistance in NSCLC/ADC cells. All-trans retinoic acid (ATRA) has been shown to be a potential targeted therapy against CSCs due to its ability to inhibit ALDH1A1 activity. We therefore investigated whether ATRA could circumvent the resistance to improve the response to gefitinib in NSCLC/ADC cells. METHODS: Treatment of NSCLC/ADC A549 and H1650 cells with gefitinib enriched the gefitinib surviving cells (GSCs). The expression of ALDH1A1 and CD44 and the IC50 values for gefitinib were determined by flow cytometry (FCM) and crystal violet assay in GSCs and ATRA-treated GSCs, respectively. Using DEAB as the positive control, direct inhibitory effect of ATRA on ALDH1A1 activity was determined by ALDEFLUOR assay, RESULTS: GSCs showed higher expression of ALDH1A1 and CD44 and IC50 values for gefitinib than their respective parental cells, suggesting that gefitinib can lead to propagation of CSC-enriched gefitinib-resistant cells. Treatment with ATRA was found to significantly reduce the increased expression of ALDH1A1 and CD44 and the IC50 values for gefitinib in A549GSC and H1650GSC cells, and ATRA could directly inhibit active ALDH1A1 as compared to DEAB. CONCLUSION: Our findings suggest that combination treatment with ATRA prevents gefitinib-induced enrichment of ALDH1A1bright/CD44high CSCs and enhances gefitinib-induced growth inhibition of NSCLC/ADC cells.

Molecular Characteristics of Methicillin-Resistant Staphylococcus epidermidis on the Abdominal Skin of Females before Laparotomy.

Staphylococcus epidermidis, especially methicillin-resistant strains, may be the source of surgical site infections and may be a reservoir of staphylococcal cassette chromosome mec (SCCmec) for S. aureus. The aim of this study was to investigate the prevalence of methicillin-resistant S. epidermidis (MRSE) on the abdominal skin of females before laparotomy and determine the molecular characteristics and antimicrobial susceptibility patterns of these isolates. MRSE was found in 54 of 157 isolates based on mecA gene detection, and there was no difference in icaA gene carriage rate between MRSE and methicillin-susceptible S. epidermidis (MSSE) isolates. Antimicrobial susceptibility profiles were determined by broth microdilution antimicrobial susceptibility testing according to the latest CLSI manuals. All MRSE isolates had unfavorable antimicrobial susceptibility patterns. Twenty-three MRSE strains (42.6%) were multi-drug resistant. SCCmec typing and pulsed field gel electrophoresis (PFGE) typing was performed. Thirty-nine (72.2%) had a single SCCmec type, whereas 1.9% had two types. Fourteen strains (25.9%) were non-typeable (NT). The most frequent MRSE genotype was SCCmec type IVa. High diversity with PFGE patterns was obtained for MRSE, and there were no isolates exhibiting identical pulsotype. The results confirm that methicillin-resistant strains are frequently present among S. epidermidis on the abdominal skin of females before laparotomy. Moreover, resistance profiles seem to have no association with the SCCmec types or PFGE types for most common antibiotics.

Anti-Trichomonas vaginalis properties of the oil of Amomum tsao-ko and its major component, geraniol.

CONTEXT: Trichomonosis, caused by the flagellate protozoan Trichomonas vaginalis, is the most common non-viral sexually transmitted disease (STD) and 5-nitroimidazole drugs are used for the treatment. However, a growing number of T. vaginalis isolates are resistant to these drugs, which make it becomes an urgent issue. OBJECTIVE: The current study was designed to evaluate the anti-T. vaginalis activity of the essential oil from A. tsao-ko used in traditional Chinese medicine and as a spice and its main component, geraniol. MATERIALS AND METHODS: The anti-T. vaginalis activities of A. tsao-ko essential oil and geraniol were evaluated by the minimum lethal concentration (MLC) and 50% inhibitory concentration (IC50) in vitro. The morphological changes of T. vaginalis were observed by transmission electron microscopy (TEM). Additionally, sub-MLC concentration treatment with sub-MLC A. tsao-ko essential oil and geraniol was also performed. RESULTS: This study shows that MLC/IC50 of A. tsao-ko essential oil was 44.97 µg/ml/22.49 µg/ml for T. vaginalis isolate Tv1, and 89.93 µg/ml/44.97 µg/ml for T. vaginalis isolate Tv2. Those of geraniol were 342.96 µg/ml/171.48 µg/ml, respectively. After A. tsao-ko essential oil or geraniol treatment, obvious similar morphological changes of T. vaginalis were observed by TEM: the nuclear membrane was damaged, nuclei were dissolved, and the chromatin was accumulated; in the cytoplasm, numerous vacuoles appeared, rough endoplasmic reticulum dilated, the number of ribosomes were reduced, organelles disintegrated, the cell membrane was partially damaged, with cytoplasmic leakage, and cell disintegration was observed. The action time did not increase the effect of A. tsao-ko essential oil or geraniol against T. vaginalis, as no significant difference was observed after sub-MLC concentration treatment for 1, 3, and 5 h with A. tsao-ko essential oil and geraniol. DISCUSSION AND CONCLUSION: The study describes the first report on the activity and morphological changes of A. tsao-ko essential oil and geraniol against T. vaginalis. The results obtained herein presented new opportunities for antitrichomonal drugs.

[Mycoplasma hominis symbiosis and Trichomonas vaginalis metronidazole resistance].

OBJECTIVE: To investigate the relation of Mycoplasma hominis symbiosis and the resistance of Trichomonas vaginalis to metronidazole. METHODS: From November 2010 to July 2011, 160 isolates of T. vaginalis were collected from the genital tract secretion of gynecological out-patients at the Sichuan Provincial Hospital for Women and Children. The minimum lethal concentration (MLC) to metronidazole of these isolates was determined by an in vitro sensitivity assay with different concentration gradients of metronidazole (from 1 to 1 024 microg/ml), and M. hominis DNA in T. vaginalis was detected by polymerase chain reaction (PCR) technique with specific 16S rRNA primers. After clearance of M. hominis from the parasites by 32 microg/ml doxycycline, MLC was determined and compared with that before clearance. RESULTS: MLC of metronidazole in T. vaginalis ranged from 1 to 256 microg/ml, with 61.3% isolates (98/160) ranging from 1 to 8 microg/ml, 26.3% isolates (42/160) ranging from 16 to 32 microg/ml, and 12.5% isolates (20/160) ranging from 64 to 256 microg/ml. 61 isolates were PCR positive for M. hominis DNA in the 160 isolates of T. vaginalis. The M. hominis DNA positive rate was significantly higher in the T. vaginalis isolates with higher MLC than those isolates with lower MLC (P<0.01). However, when M. hominis was cleared by doxycycline from 8 isolates among the 61 ones, no change was observed in sensitivity of the isolates to metronidazole. CONCLUSION: M. hominis symbiosis might be associated with the metronidazole-resistance of T. vaginalis. However, it needs direct evidence.

Identification and distribution of the clinical isolates of imipenem-resistant Pseudomonas aeruginosa carrying metallo-beta-lactamase and/or class 1 integron genes.

To investigate the distribution of the genes of two major metallo-beta-lactamases (MBL; i.e., IMP and VIM) and class 1 integrons (intI) in the clinical imipenem-resistant Pseudomonas aeruginosa, a total of 65 isolates, from a university hospital in Sichuan between December 2004 and April 2005 were screened for MBL genes by PCR using primers specific for bla(IMP-1), bla(VIM) and bla(VIM-2) genes. The MBL-positive isolates were further assessed for class 1 integrons by PCR using specific primers. The nucleotide sequences of several PCR products were also determined. The results revealed that the bla(VIM) gene was found in 81.5% (53/65) of all isolates, bla(VIM-2) gene was found in only 1 isolate and the intI gene was observed in 45.3% (24/53) of bla(VIM)-positive isolates. One isolate carried simultaneously both bla(IMP-1) and intI genes, and to the best of our knowledge this is the first report of such isolate in southwest China. These observations highlight that the genes for VIM beta-lactamase and class 1 integrons were predominantly present among the imipenem-resistant P. aeruginosa tested, confirming the current widespread threat of imipenem-resistant, integron-borne P. aeruginosa.

Identification and Distribution of the Clinical Isolates of Imipenem-resistant Pseudomonas aeruginosa Carrying Metallo-β-lactamase and/or Class 1 Integron Genes / 华中科技大学学报（医学）（英德文版）

To investigate the distribution of the genes of two major metallo-β-lactamases (MBL; i.e., IMP and VIM) and class 1 integrons (intI) in the clinical imipenem-resistant Pseudomonas aeruginosa, a total of 65 isolates, from a university hospital in Sichuan between December 2004 and April 2005 were screened for MBL genes by PCR using primers specific for blaIMP-1, blaVIM and blaVIM-2 genes. The MBL-positive isolates were further assessed for class 1 integrons by PCR using specific primers. The nucleotide sequences of several PCR products were also determined. The results revealed that the blaVIM gene was found in 81.5% (53/65) of all isolates, bla<VIM-2> gene was found in only 1 isolate and the intI gene was observed in 45.3% (24/53) of blaVIM-positive isolates. One isolate carried simultaneously both blaIMP-1 and intI genes, and to the best of our knowledge this is the first report of such isolate in southwest China. These observations highlight that the genes for VIM β-lactamase and class 1 integrons were predominantly present among the imipenem-resistant P. aeruginosa tested, confirming the current widespread threat of imipenem-resistant, integron-borne P. aeruginosa.