This study primarily aimed to investigate the prevalence of human papillomavirus (HPV) and other common pathogens of sexually transmitted infections (STIs) in spermatozoa of infertile men and their effects on semen parameters. These pathogens included Ureaplasma urealyticum, Ureaplasma parvum, Chlamydia trachomatis, Mycoplasma genitalium , herpes simplex virus 2, Neisseria gonorrhoeae, Enterococcus faecalis, Streptococcus agalactiae, Pseudomonas aeruginosa , and Staphylococcus aureus . A total of 1951 men of infertile couples were recruited between 23 March 2023, and 17 May 2023, at the Department of Reproductive Medicine of The First People's Hospital of Yunnan Province (Kunming, China). Multiplex polymerase chain reaction and capillary electrophoresis were used for HPV genotyping. Polymerase chain reaction and electrophoresis were also used to detect the presence of other STIs. The overall prevalence of HPV infection was 12.4%. The top five prevalent HPV subtypes were types 56, 52, 43, 16, and 53 among those tested positive for HPV. Other common infections with high prevalence rates were Ureaplasma urealyticum (28.3%), Ureaplasma parvum (20.4%), and Enterococcus faecalis (9.5%). The prevalence rates of HPV coinfection with Ureaplasma urealyticum, Ureaplasma parvum, Chlamydia trachomatis, Mycoplasma genitalium , herpes simplex virus 2, Neisseria gonorrhoeae, Enterococcus faecalis, Streptococcus agalactiae , and Staphylococcus aureus were 24.8%, 25.4%, 10.6%, 6.4%, 2.4%, 7.9%, 5.9%, 0.9%, and 1.3%, respectively. The semen volume and total sperm count were greatly decreased by HPV infection alone. Coinfection with HPV and Ureaplasma urealyticum significantly reduced sperm motility and viability. Our study shows that coinfection with STIs is highly prevalent in the semen of infertile men and that coinfection with pathogens can seriously affect semen parameters, emphasizing the necessity of semen screening for STIs.
Humans ; Male ; Infertility, Male/epidemiology* ; Coinfection/microbiology* ; Papillomavirus Infections/virology* ; Adult ; Sexually Transmitted Diseases/complications* ; China/epidemiology* ; Staphylococcus aureus/isolation & purification* ; Chlamydia trachomatis/isolation & purification* ; Prevalence ; Mycoplasma genitalium/isolation & purification* ; Ureaplasma urealyticum/isolation & purification* ; Neisseria gonorrhoeae/isolation & purification* ; Enterococcus faecalis/isolation & purification* ; Streptococcus agalactiae/isolation & purification* ; Herpesvirus 2, Human/genetics* ; Pseudomonas aeruginosa/isolation & purification* ; Semen/virology* ; Sperm Motility ; Spermatozoa/microbiology* ; Human Papillomavirus Viruses

Endosomes are characterized by the presence of various phosphoinositides that are essential for defining the membrane properties. However, the interplay between endosomal phosphoinositides metabolism and innate immunity is yet to be fully understood. Here, our findings highlight the evolutionary continuity of RAB-10/Rab10's involvement in regulating innate immunity. Upon infection of Caenorhabditis elegans with Pseudomonas aeruginosa, an increase in RAB-10 activity was observed in the intestine. Conversely, when RAB-10 was absent, the intestinal diacylglycerols (DAGs) decreased, and the animal's response to the pathogen was impaired. Further research revealed that UNC-16/JIP3 acts as an RAB-10 effector, facilitating the recruitment of phospholipase EGL-8 to endosomes. This leads to a decrease in endosomal phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2) and an elevation of DAGs, as well as the activation of the PMK-1/p38 MAPK innate immune pathway. It is noteworthy that the dimerization of UNC-16 is a prerequisite for its interaction with RAB-10(GTP) and the recruitment of EGL-8. Moreover, we ascertained that the rise in RAB-10 activity, due to infection, was attributed to the augmented expression of LET-413/Erbin, and the nuclear receptor NHR-25/NR5A1/2 was determined to be indispensable for this increase. Hence, this study illuminates the significance of endosomal PI(4,5)P2 catabolism in boosting innate immunity and outlines an NHR-25-mediated mechanism for pathogen detection in intestinal epithelia.
Animals ; Caenorhabditis elegans/genetics* ; Endosomes/immunology* ; Caenorhabditis elegans Proteins/immunology* ; Phosphatidylinositol 4,5-Diphosphate/immunology* ; Immunity, Innate ; Pseudomonas aeruginosa/immunology* ; rab GTP-Binding Proteins/genetics* ; Diglycerides/metabolism*

OBJECTIVE: Pseudomonas aeruginosa( P. aeruginosa) is a prevalent pathogenic bacterium involved in meningitis; however, the virulence factors contributing to this disease remain poorly understood. METHODS: The virulence of the P. aeruginosa A584, isolated from meningitis samples, was evaluated by constructing in vitro blood-brain barrier and in vivo systemic infection models. qPCR, whole-genome sequencing, and drug efflux assays of A584 were performed to analyze the virulence factors. RESULTS: Genomic sequencing showed that A584 formed a phylogenetic cluster with the reference strains NY7610, DDRC3, Pa58, and Pa124. Its genome includes abundant virulence factors, such as hemolysin, the Type IV secretion system, and pyoverdine. A584 is a multidrug-resistant strain, and its wide-spectrum resistance is associated with enhanced drug efflux. Moreover, this strain caused significantly more severe damage to the blood-brain barrier than the standard strain, PAO1. qPCR assays further revealed the downregulation of the blood-brain barrier-associated proteins Claudin-5 and Occludin by A584. During systemic infection, A584 exhibited a higher capacity of brain colonization than PAO1 (37.1 × 10 ⁶ CFU/g brain versus 2.5 × 10 ⁶ CFU/g brain), leading to higher levels of the pro-inflammatory factors IL-1β and TNF-α. CONCLUSION This study sheds light on the virulence factors of P. aeruginosa involved in meningitis.
Pseudomonas aeruginosa/genetics* ; Virulence Factors/metabolism* ; Animals ; Virulence ; Mice ; Pseudomonas Infections/microbiology* ; Blood-Brain Barrier/microbiology* ; Humans ; Female

2-substituted quinolines are the building blocks for the synthesis of natural products and pharmaceuticals. In comparison with classical methods, dehydroaromatization of 2-substituted-1,2,3,4-tetrahydroquinolines has emerged in recent years as an efficient and straightforward method to synthesize quinolines due to its high atom economy and sustainability. However, existing chemical methods need transition metal catalysts and harsh reaction conditions. Biocatalysis with high efficiency, high selectivity, and mild reaction conditions has become an important method of organic synthesis. We mined a strain Pseudomonas monteilii ZMU-T06 capable of producing monoamine oxidase for the dehydroaromatization of 2-substituted-1,2,3,4-tetrahydroquinolines to synthesize 2-substituted quinolines (8 substrates, yields of 45.7%-48.4%) and then hypothesized the catalytic mechanism, providing a new method for green synthesis of 2-substituted quinolines.
Quinolines/chemistry* ; Pseudomonas/classification* ; Oxidation-Reduction ; Monoamine Oxidase/biosynthesis* ; Biocatalysis

To develop biocontrol agents for the control of kiwifruit bacterial canker, we isolated a strain CXG2-5 with inhibitory activity against Pseudomonas syringae pv. actinidiae (Psa), the pathogen of kiwifruit bacterial canker, from the rhizosphere soil of kiwifruit by the plate confrontation test. The strain was identified by morphological observation, physiological and biochemical tests, and molecular biological methods. The indoor control efficacy of the strain was determined by the inoculation of the strain into detached branches with wounds and into leaf discs by vacuum infiltration. The ability of the strain to expand and colonize leaf veins was determined by fluorescent labeling and scanning electron microscopy. Subsequently, the strain was prepared into microcapsules, the field control efficacy of which was evaluated. The strain CXG2-5 was identified as Pseudomonas benzenivorans. It demonstrated good antagonistic activity against Psa, with an inhibition zone diameter of 22 mm and an inhibition rate of 72.7%. The preventive effects of the strain on kiwifruit bacterial canker were better than the therapeutic effects on both detached branches and leaves, with the preventive effects reaching 65% and 92.4%, respectively. The control effect of microcapsules of this strain in the field reached 60.89%, which was slightly lower than that of 20% kasugamycin and higher than that of Bacillus subtilis wettable powder. In conclusion, strain CXG2-5 serves as a candidate for the control of kiwifruit bacterial canker, and the prepared microcapsules have good value for development and application.
Actinidia/microbiology* ; Plant Diseases/prevention & control* ; Pseudomonas syringae ; Pseudomonas/isolation & purification* ; Capsules ; Antibiosis ; Biological Control Agents ; Pest Control, Biological/methods*

Objective: To report a case of acute postoperative endophthalmitis following cataract surgery due to Pseudomonas stutzeri in a healthy elderly male. Methods: This is a case report. Results: A non-hypertensive, non-diabetic male in his late 60s consulted due to eye pain and blurred vision 5 days after an uncomplicated extracapsular cataract extraction with posterior chamber intraocular lens implantation (PCIOL) on his left eye. On examination, the visual acuity was light perception. Slit-lamp examination showed ciliary injection, conjunctival congestion, mild corneal edema with Descemet membrane folds, hazy anterior chamber with fibrin and a 2-millimeter hypopyon, and a visible PCIOL. IOP was 10 mmHg with no leak on Seidel’s test, and there was poor view of the fundus. B-scan ultrasonography showed findings consistent with endophthalmitis. He was given topical, intravitreal, and systemic antibiotics, and emergency vitrectomy was done. The vitreous sample culture revealed Pseudomonas stutzeri. Despite aggressive medical and surgical management, vision loss was not prevented. Conclusion Acute postoperative endophthalmitis from Pseudomonas stutzeri is rare; if not recognized and treated promptly, this complication has devastating outcomes. It may present with a fulminant course regardless of the associated risks for infection. Prevention, early recognition, and timely management can prevent unfavorable visual outcomes.
Endophthalmitis ; Pseudomonas stutzeri

Pyocin S2 and S4 in Pseudomonas aeruginosa use the same uptake channels as the pyoverdine does in bacteria, indicating a possible connection between them. In this study, we characterized the single bacterial gene expression distribution of three S-type pyocins (Pys2, PA3866, and PyoS5) and examined the impact of pyocin S2 on bacterial uptake of pyoverdine. The findings demonstrated that the expression of the S-type pyocin genes was highly differentiated in bacterial population under DNAdamage stress. Moreover, exogenous addition of pyocin S2 reduces the bacterial uptake of pyoverdine so that the presence of pyocin S2 prevents the uptake of environmental pyoverdine by non-pyoverdine synthesizing 'cheaters', thereby reducing their resistance to oxidative stress. Furthermore, we discovered that overexpression of the SOS response regulator PrtN in bacteria significantly decreased the expression of genes involved in the synthesis of pyoverdine, significantly decreasing the overall synthesis and exocytosis of pyoverdine. These findings imply a connection between the function of the iron absorption system and the SOS stress response mechanism in bacteria.
Pyocins/metabolism* ; Pseudomonas aeruginosa/metabolism*

Manipulation of genes, including knock-out or knock-in, replacement of gene elements (such as promoters), fusion with a fluorescent protein gene, and construction of in situ gene reporter, is required in most of the biotechnological laboratories. The widely used gene manipulating methods based on two-step allelic exchange are cumbersome in terms of constructing plasmids, transforming and screening. In addition, the efficiency of using this method for long fragment knockout is low. To simplify the process of gene manipulation, we constructed a minimized integrative vector pln2. When a gene needs to be inactivated, an internal fragment of the target gene (non-frameshift) is cloned into the pln2 plasmid. Once the single-crossover recombination between genome and the constructed plasmid occurs, the endogenous gene is segmented by the plasmid backbone and thus inactivated. We developed a toolbox based on pln2 that can be used for different genomic operation mentioned above. With the help of this toolbox, we successfully knocked out large fragments of 20-270 kb.
Genetic Vectors/genetics* ; Pseudomonas aeruginosa/genetics* ; Plasmids/genetics* ; Promoter Regions, Genetic ; Genome

Lysis is a common functional module in synthetic biology and is widely used in genetic circuit design. Lysis could be achieved by inducing expression of lysis cassettes originated from phages. However, detailed characterization of lysis cassettes hasn't been reported yet. Here, we first adopted arabinose- and rhamnose-inducible systems to develop inducible expression of five lysis cassettes (S₁₀₅, A52G, C51S S76C, LKD, LUZ) in Escherichia coli Top10. By measuring OD₆₀₀, we characterized the lysis behavior of strains harboring different lysis cassettes. These strains were harvested at different growth stages, induced with different concentrations of chemical inducers, or contained plasmids with different copy numbers. We found that although all five lysis cassettes could induce bacterial lysis in Top10, lysis behaviors differed a lot at various conditions. We further found that due to the difference in background expression levels between strain Top10 and Pseudomonas aeruginosa PAO1, it was hard to construct inducible lysis systems in strain PAO1. The lysis cassette controlled by rhamnose-inducible system was finally inserted into the chromosome of strain PAO1 to construct lysis strains after careful screen. The results indicated that LUZ and LKD were more effective in strain PAO1 than S₁₀₅, A52G and C51S S76C. At last, we constructed an engineered bacteria Q16 using an optogenetic module BphS and the lysis cassette LUZ. The engineered strain was capable of adhering to target surface and achieving light-induced lysis by tuning the strength of ribosome binding sites (RBSs), showing great potential in surface modification.
Rhamnose/pharmacology* ; Plasmids/genetics* ; Pseudomonas aeruginosa ; Escherichia coli/metabolism*

OBJECTIVE: To investigate the distribution and drug sensitivity of pathogenic bacteria isolated from patients in hematology department, in order to provide evidence for rational use of antibiotics in clinic. METHODS: The distribution of pathogenic bacteria and drug sensitivity data of patients in the hematology department of The First Affiliated Hospital of Nanjing Medical University from 2015 to 2020 were retrospectively analyzed, and the pathogens isolated from different specimen types were compared. RESULTS: A total of 2 029 strains of pathogenic bacteria were isolated from 1 501 patients in the hematology department from 2015 to 2020, and 62.2% of which were Gram-negative bacilli, mainly Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Stenotrophomonas maltophilia and Acinetobacter baumannii. Gram-positive coccus accounted for 18.8%, mainly Coagulase-negative staphylococcus (CoNS) and Staphylococcus aureus. Fungi (17.4%) were mainly candida. The 2 029 strains were mainly isolated from respiratory tract (35.1%), blood (31.8%) and urine (19.2%) specimens. Gram-negative bacilli were the main pathogenic bacteria in different specimen types (>60%). K. pneumoniae, S. maltophilia and A. baumannii were the most common pathogens in respiratory specimens, E. coli, CoNS, K. pneumoniae and P. aeruginosa were common in blood samples, and E. coli and Enterococcus were most common in urine samples. Enterobacteriaceae had the highest susceptibility to amikacin and carbapenems (>90.0%), followed by piperacillin/tazobactam. P. aeruginosa strains had high sensitivity to antibiotics except aztreonam (<50.0%). The susceptibility of A. baumannii to multiple antibiotics was less than 70.0%. The antimicrobial resistance rates of E. coli and K. pneumoniae in respiratory tract specimens were higher than those in blood specimens and urine specimens. CONCLUSION Gram-negative bacilli are the main pathogenic bacteria isolated from patients in hematology department. The distribution of pathogens is different in different types of specimens, and the sensitivity of each strain to antibiotics is different. The rational use of antibiotics should be based on different parts of infection to prevent the occurrence of drug resistance.
Humans ; Escherichia coli ; Retrospective Studies ; Bacteria ; Anti-Bacterial Agents/therapeutic use* ; Gram-Negative Bacteria ; Drug Resistance ; Pseudomonas aeruginosa ; Hematology