Objective　To detect clustered regularly interspaced short palindromic repeats (CRISPR) in Listeria monocytogenes, and analyze the structure and homology of CRISPR loci. Methods Totally 34 strains of Listeria monocytogenes isolated in our laboratory were identified, PCR amplified and sequenced. The repeat sequence structure and spacer sequence homology in CRISPR loci were analyzed by bioinformatics software. Results A total of 7 CRISPR loci were detected in 34 strains. The mutation rate of the first 2 and last 2 bases of the Repeat sequence of CRISPR loci was higher, while the mutation rate of the middle part was lower. Seven CRISPR sites form eight CRISPR structural types, among which the Repeat sequences of CRISPR1 and CRISPR2 are relatively conserved, while the Repeat sequences of CRISPR1 and CRISPR5 can form dumbbell shaped secondary structures. The number of Spacer sequences contained in each CRISPR site ranges from 2 to 15, with an average of 2.43. The 136 Spacer sequences detected were not only homologous to Listeria plasmids and bacteriophages, but also homologous to uncultured virus sequences, staphylococcal bacteriophages, and Listeria innocua. The same CRISPR genotype did not show large-scale clustering, but some strains in the same year were in the same evolutionary cluster with close genetic relationships. Conclusion The CRISPR structure of Listeria monocytogenes in this study exhibits high specificity, and its homology with bacteriophages provides a theoretical basis for the application of bacteriophages in the control and prevention of Listeria monocytogenes.

Human NAD(P)H: quinone oxidoreductase 1 (NQO1) is a flavoenzyme expressed at high levels in multiple solid tumors, making it an attractive target for anticancer drugs. Bioactivatable drugs targeting NQO1, such as β-lapachone (β-lap), are currently in clinical trials for the treatment of cancer. β-Lap selectively kills NQO1-positive (NQO1⁺) cancer cells by inducing reactive oxygen species (ROS) via catalytic activation of NQO1. In this study, we demonstrated that cryptotanshinone (CTS), a naturally occurring compound, induces NQO1-dependent necrosis without affecting NQO1 activity. CTS selectively kills NQO1⁺ cancer cells by inducing NQO1-dependent necrosis. Interestingly, CTS directly binds to NQO1 but does not activate its catalytic activity. In addition, CTS enables activation of JNK1/2 and PARP, accumulation of iron and Ca²⁺, and depletion of ATP and NAD⁺. Furthermore, CTS selectively suppressed tumor growth in the NQO1⁺ xenograft models, which was reversed by NQO1 inhibitor and NQO1 shRNA. In conclusion, CTS induces NQO1-dependent necrosis via the JNK1/2/iron/PARP/NAD⁺/Ca²⁺ signaling pathway. This study demonstrates the non-enzymatic function of NQO1 in inducing cell death and provides new avenues for the design and development of NQO1-targeted anticancer drugs.

Objective:To explore the effect of emotional imagery desensitization therapy on posttraumatic stress disorder among school- age children with fracture, to alleviate the adverse psychological state of the children.Methods:By a randomized controlled trials, a total of 78 school- age children with fracture at the Affiliated Hospital of Jining Medical University from July 2021 to July 2023 were divided into experimental group and control group according to the admission time, with 39 cases in each group. Both groups carried out routine care, the control group was given routine psychological intervention, the experimental group implemented emotional imagery desensitization therapy. Before and after 3 months of intervention, the effects was assessed by Post-Traumatic Stress Disorder Self-rating Scale (PTSD-SS) and Medical Coping Modes Questionnaire (MCMQ), respectively.Results:During the research process, there were 2 cases of dropout in the control group due to disconnection, and 4 cases in the experimental group due to disconnection and voluntary withdrawal. 37 children in the control group [20 males and 17 females, age (9.27 ± 1.62) years], and 35 children in the experimental group [21 males and 14 females, age (9.57 ± 1.63) years] completed the intervention. Before intervention, the difference of PTSD-SS and MCMQ scores between two groups was no statistically significant (all P>0.05). After intervention, the subjective assessment, re-experiencing, avoidance symptoms, hypervigilance, functional impairment factor scores and total PTSD-SS scores were (1.51 ± 1.06), (10.74 ± 3.30), (10.86 ± 2.93), (11.94 ± 3.18), (2.31 ± 1.30), (37.37 ± 6.37) in the experimental group, lower than in the control group (2.08 ± 0.80), (12.92 ± 3.63), (12.73 ± 2.99), (14.65 ± 4.23), (2.97 ± 0.87), (45.35 ± 5.86), the differences were statistically significant ( t=2.53-5.54, all P<0.05). After intervention, the confrontation subscale scores were (23.49 ± 2.48), higher than in the control group (21.89 ± 2.38), the avoidance, acceptance- resignation subscale were (10.31 ± 1.89) and (6.83 ± 1.98), lower than in the control group (11.57 ± 1.79) and (7.86 ± 2.12), the differences were statistically significant ( t=2.79, 2.89, 2.14, all P<0.05). Conclusions:Emotional imagery desensitization therapy can effectively alleviate post- traumatic stress disorder and promote positive coping mode in school-age children with fracture.

Objective To investigate the effect of adenotonsillectomy on immune function and sleep structure in children with obstructive sleep apnea-hypopnea syndrome(OSAHS).Methods A total of 94 children with OSAHS treated at Children's Hospital Affiliated to Zhengzhou University from November 2020 to June 2022 were selected as the observation group,and another 80 healthy children who underwent physical examinations at the same hospital during the same period were selected as the control group.Children in the observation group underwent bilateral tonsillectomy combined with endoscopic adenoidectomy.Changes in the sleep structure of children in the observation group were detected before surgery and at discharge by using a multi-channel sleep monitoring system.Venous blood samples were collected from children in the observation group before surgery,one month after surgery,and six months after surgery,while venous blood samples of children in the control group were collected on the day of physical examination.The serum levels of IgA,IgG and IgM were measured by using immunoturbidimetry,the percentages of CD3+,CD4+and CD8+in the plasma were measured by using flow cytometry,and the CD4+/CD8+ratio was calculated.Results Compared with before surgery,the proportion of non-rapid eye movement sleep(NREM)1,apnea-hypopnea index and obstructive apnea-hypopnea index of children in the observation group decreased at discharge,the proportions of NREM2,NREM3 and rapid eye movement sleep(REM),sleep efficiency and the lowest oxygen saturation increased,and REM time extended(P＜0.05).The preoperative serum levels of IgA,IgG and IgM in the observation group were significantly higher than those in the control group(P＜0.05).One month after surgery,the serum levels of IgA,IgG and IgM in the observation group were significantly lower than those in the control group(P＜0.05).There was no statistically significant difference in the serum levels of IgA,IgG and IgM between the observation group and the control group at six months after surgery(P＞0.05).One and six months after surgery,the serum levels of IgA,IgG and IgM in the observation group were significantly lower than those before surgery(P＜0.05),and the serum levels of IgA,IgG and IgM at six months after surgery were significantly higher than those one month after surgery(P＜0.05).The preoperative CD3+and CD4+percentages and the CD4+/CD8+ratio in the observation group were significantly lower than those in the control group,while the CD8+percentage was significantly higher than that in the control group(P＜0.05).One month after surgery,the CD3+and CD4+percentages and the CD4+/CD8+ratio in the observation group were significantly lower than those in the control group,while the CD8+percentage was significantly higher than that in the control group(P＜0.05).There was no statistically significant difference in the CD3+,CD4+and CD8+percentages and the CD4+/CD8+ratio between the observation group and the control group at six months after surgery(P＞0.05).One month after surgery,the CD3+and CD4+percentages and the CD4+/CD8+ratio in the observation group were significantly lower than those before surgery,while the CD8+percentage was significantly higher than that before surgery(P＜0.05);six months after surgery,the CD3+and CD4+percentages and the CD4+/CD8+ratio in the observation group were significantly higher than those before surgery,while the CD8+percentage was significantly lower than that before surgery(P＜0.05).The CD3+and CD4+percentages and the CD4+/CD8+ratio at six months after surgery were significantly higher than those at one month after surgery,while the CD8+percentage was significantly lower than that at one month after surgery in the observation group(P＜0.05).Conclusion Adenotonsillectomy can effectively prolong the REM time of OSAHS children and improve their sleep efficiency,sleep structure and the immune regulation function of the body.The immune function of the body decreases briefly after adenotonsillectomy,and then gradually returns to the normal level.

Objective: To investigate the incidence and influencing factors of falls among the elderly in Ningbo City, Zhejiang Province, so as to provide the basis for developing effective prevention strategies. Methods: The residents aged 60 years and above in Haishu District and Yuyao City of Ningbo City were selected by the multi-stage cluster random sampling method from June to October 2022. Demographic information, fall incidence in the past year, history of disease and self-rated health were collected through questionnaire surveys. Incidence of falls was descriptively analyzed, and factors affecting falls were identified using a multivariable logistic regression model stratified by gender and age. Results: A total of 1 275 elderly people were surveyed, including 635 men and 640 women. The median age was 72.00 (interquartile range, 13.00) years. In the past year, 158 residents fell, accounting for 12.39%. Additionally, 14 individuals experienced two or more falls, accounting for 8.86%. The incidence of falls was 14.69% in women, which was higher than the 10.08% in men (P<0.05). The incidence of falls was 14.86% in the elderly over 70 years, which was higher than the 9.39% in those aged 60 to 70 years (P<0.05). Multivariable logistic regression showed that the educational level (primary school and above, OR=0.501, 95%CI: 0.301-0.836), heart disease (present, OR=1.996, 95%CI: 1.076-3.703), and self-rated health status (good, OR=0.529, 95%CI: 0.319-0.875) were factors affecting falls in women; educational level (primary school and above, OR=0.514, 95%CI: 0.285-0.928) and self-rated health status (good, OR=0.456, 95%CI: 0.253-0.824) were factors affecting falls in residents aged 60 to 70 years. Conclusion Fall risk among the elderly is associated with gender, age, heart disease, educational level and self-rated health status, and the influencing factors for falls vary in different genders and ages.

Objective:To analyze the clinical characteristics, treatment, and prognosis of ectopic bronchogenic cysts in children.Methods:A retrospective analysis was conducted on the data including the clinical characteristics, auxiliary examination and treatment of 21 children with ectopic bronchogenic cysts diagnosed pathologically at Children′s Hospital Affiliated to Zhengzhou University from July 2015 to December 2023. There were 16 males and 5 females, with a male-female ratio of 3.2∶1, and the age ranged from 4 days to 8 years old (median age 2 years and 8 months).Results:Among the 21 cases of ectopic bronchogenic cysts, 11 cases were found in the pharynx, with symptoms including dyspnea (4 cases), snoring during sleep (3 cases), and choking on milk(4 cases).Ten cases were found in the head, neck or anterior chest, 5 of these cases had infection history, and 5 showed progressive mass growth.Imaging and endoscopy showed 9 patients underwent preoperative color ultrasonography revealed cystic masses with well-defined boundaries. CT examination was performed on 13 patients, which showed round or nearly round masses with homogeneous density, smooth margins, and regular cyst walls. CT attenuation values ranged from 2 to 52 Hounsfield Units (HU). Four cystic lesions were assessed via MRI, 3 cases demonstrated long T1 and long T2 signals, while 1 case had a slight short T1 and long T2 signal, with high signal intensity on fat-suppressed images. Eleven cases of pharyngopharyngeal cysts were examined by electronic nasopharyngoscopy. The cysts appeared as spherical or ovoid masses with smooth surfaces, close to or slightly light in color with the surrounding tissue, with one cyst presenting with a bluish blue in the oropharynx. All 11 pharyngeal cysts were excised using low-temperature plasma under general anesthesia and intubation assisted by a nasal endoscope. The cysts were pulled and excised as completely as possible.Ten cases of neck and anterior chest cysts were completely excised. Postoperative histopathology confirmed bronchogenic cyst. Twenty-one children were followed up postoperatively for 4 months to 7 years without recurrence, except for 1 patient who was lost to follow-up.Conclusions:Ectopic bronchogenic cysts are uncommon and lack of typical imaging and clinical features.Combination of ultrasonography, CT and MRI is recommended for cases occuered in neck and anterior chest, while electronic nasopharyngoscopy complements pharyngeal evaluations. Surgical intervention is the preferred treatment choice for this disease.

Diabetic retinopathy(DR)is one of the most common retinal complications of diabetes could cause irreversible loss of central vision in the working-age population. Current studies showed that systemic risk factors, inflammatory response, and oxidative stress played a central role in the development of DR. Although traditional sequencing methods have provided valuable insights into the pathogenesis of DR, offering crucial guidance for clinical diagnosis and treatment, they still possess certain limitations. In recent years, the emerging single-cell RNA sequencing technology(scRNA-seq)has enabled precise analysis of mRNA transcriptomes at the single-cell level. This technique accurately identifies novel cell subtypes in retinal diseases, detects rare cells, and reveals intercellular heterogeneity. It contributes to elucidating the pathogenesis and development of retinal diseases, and facilitates exploration of gene regulatory relationships associated with these disorders to provide valuable insights for future precision medicine. This article reviews the technology of single-cell sequencing and its application in DR research. It also explores the mechanisms of different types of cells associated with DR, aiming to enhance the utilization of scRNA-seq in DR research and identify potential therapeutic targets to improve clinical diagnosis and treatment of DR.

IKBKG is the essential modulator for nuclear factor-κB(NF-κB) signaling pathway, and mutations within this gene can lead to anhidrotic ectodermal dysplasia and immunodeficiency (EDA-ID). Here we report a male patient, who presented with mild frontal bossing, sparse hair, skin pigmentation, conical teeth, and recurrent infections involving bacteria, fungi, and viruses after one month of age, together with hypogammaglobulinemia. These symptoms were consistent with the phenotype of EDA-ID. Genetic analysis revealed a hemizygous mutation c.1249T > G (p.Cys417Gly) in exon 10 of the IKBKG gene in the patient. The mother of the patient was identified as heterozygous carriers of the same mutation. Immunological assessment revealed a significant reduction in memory B cells. The patient showed no skewed TCR diversity. PHA-induced T-cell proliferation was impaired. IFN-γ secretion by Th cells was significantly reduced after PHA stimulation. IκBα degradation rate retained the same in the patient. We summarized the clinical features of the patient and conducted immunological analysis, in order to increase pediatricians′awareness of this rare disease. For boys with early-onset recurrent infections together with ectodermal dysplasia, IKBKG mutation should be considered. In addition, genetic analysis is needed to exclude pseudogene interference and functional evaluations are required.

Objective:To report the results of national surveillance on the distribution and antimicrobial resistance profile of clinical Gram-negative bacteria isolates from bloodstream infections in China in 2022.Methods:The clinical isolates of Gram-negative bacteria from blood cultures in member hospitals of national bloodstream infection Bacterial Resistant Investigation Collaborative System（BRICS）were collected during January 2022 to December 2022. Antibiotic susceptibility tests were conducted by agar dilution or broth dilution methods recommended by Clinical and Laboratory Standards Institute（CLSI）. WHONET 5.6 and SPSS 25.0 software were used to analyze the data.Results:During the study period，9 035 strains of Gram-negative bacteria were collected from 51 hospitals，of which 7 895（87.4%）were Enterobacteriaceae and 1 140（12.6%）were non-fermenting bacteria. The top 5 bacterial species were Escherichia coli（ n=4 510，49.9%）， Klebsiella pneumoniae（ n=2 340，25.9%）， Pseudomonas aeruginosa（ n=534，5.9%）， Acinetobacter baumannii complex（ n=405，4.5%）and Enterobacter cloacae（ n=327，3.6%）. The ESBLs-producing rates in Escherichia coli， Klebsiella pneumoniae and Proteus spp. were 47.1%（2 095/4 452），21.0%（427/2 033）and 41.1%（58/141），respectively. The prevalence of carbapenem-resistant Escherichia coli（CREC）and carbapenem-resistant Klebsiella pneumoniae（CRKP）were 1.3%（58/4 510）and 13.1%（307/2 340）；62.1%（36/58）and 9.8%（30/307）of CREC and CRKP were resistant to ceftazidime/avibactam combination，respectively. The prevalence of carbapenem-resistant Acinetobacter baumannii（CRAB）complex was 59.5%（241/405），while less than 5% of Acinetobacter baumannii complex was resistant to tigecycline and polymyxin B. The prevalence of carbapenem-resistant Pseudomonas aeruginosa（CRPA）was 18.4%（98/534）. There were differences in the composition ratio of Gram-negative bacteria in bloodstream infections and the prevalence of main Gram-negative bacteria resistance among different regions，with statistically significant differences in the prevalence of CRKP and CRPA（ χ2=20.489 and 20.252， P<0.001）. The prevalence of CREC，CRKP，CRPA，CRAB，ESBLs-producing Escherichia coli and Klebsiella pneumoniae were higher in provinicial hospitals than those in municipal hospitals（ χ2=11.953，81.183，10.404，5.915，12.415 and 6.459， P<0.01 or <0.05），while the prevalence of CRPA was higher in economically developed regions（per capita GDP ≥ 92 059 Yuan）than that in economically less-developed regions（per capita GDP <92 059 Yuan）（ χ2=6.240， P=0.012）. Conclusions:The proportion of Gram-negative bacteria in bloodstream infections shows an increasing trend，and Escherichia coli is ranked in the top，while the trend of CRKP decreases continuously with time. Decreasing trends are noted in ESBLs-producing Escherichia coli and Klebsiella pneumoniae. Low prevalence of carbapenem resistance in Escherichia coli and high prevalence in CRAB complex have been observed. The composition ratio and antibacterial spectrum of bloodstream infections in different regions of China are slightly different，and the proportion of main drug resistant bacteria in provincial hospitals is higher than those in municipal hospitals.

Objective:To report the results of national surveillance on the distribution and antimicrobial resistance profile of clinical Gram-positive bacteria isolates from bloodstream infections in China in 2022.Methods:The clinical isolates of Gram-positive bacteria from blood cultures in member hospitals of National Bloodstream Infection Bacterial Resistant Investigation Collaborative System（BRICS）were collected during January 2022 to December 2022. Antibiotic susceptibility tests were conducted by agar dilution or broth dilution methods recommended by Clinical and Laboratory Standards Institute（CLSI）. WHONET 5.6 and SPSS 25.0 software were used to analyze the data.Results:A total of 3 163 strains of Gram-positive pathogens were collected from 51 member units，and the top five bacteria were Staphylococcus aureus（ n=1 147，36.3%），coagulase-negative Staphylococci（ n=928，29.3%）， Enterococcus faecalis（ n=369，11.7%）， Enterococcus faecium（ n=296，9.4%）and alpha-hemolyticus Streptococci（ n=192，6.1%）. The detection rates of methicillin-resistant Staphylococcus aureus（MRSA）and methicillin-resistant coagulase-negative Staphylococci（MRCNS）were 26.4%（303/1 147）and 66.7%（619/928），respectively. No glycopeptide and daptomycin-resistant Staphylococci were detected. The sensitivity rates of Staphylococcus aureus to cefpirome，rifampin，compound sulfamethoxazole，linezolid，minocycline and tigecycline were all >95.0%. Enterococcus faecium was more prevalent than Enterococcus faecalis. The resistance rates of Enterococcus faecium to vancomycin and teicoplanin were both 0.5%（2/369），and no vancomycin-resistant Enterococcus faecium was detected. The detection rate of MRSA in southern China was significantly lower than that in other regions（ χ2=14.578， P=0.002），while the detection rate of MRCNS in northern China was significantly higher than that in other regions（ χ2=15.195， P=0.002）. The detection rates of MRSA and MRCNS in provincial hospitals were higher than those in municipal hospitals（ χ2=13.519 and 12.136， P<0.001）. The detection rates of MRSA and MRCNS in economically more advanced regions（per capita GDP≥92 059 Yuan in 2022）were higher than those in economically less advanced regions（per capita GDP<92 059 Yuan）（ χ2=9.969 and 7.606， P=0.002和0.006）. Conclusions:Among the Gram-positive pathogens causing bloodstream infections in China， Staphylococci is the most common while the MRSA incidence decreases continuously with time；the detection rate of Enterococcus faecium exceeds that of Enterococcus faecalis. The overall prevalence of vancomycin-resistant Enterococci is still at a low level. The composition ratio of Gram-positive pathogens and resistant profiles varies slightly across regions of China，with the prevalence of MRSA and MRCNS being more pronounced in provincial hospitals and areas with a per capita GDP≥92 059 yuan.