The culture of suspended Vero cells is facing difficulties such as low cell viability and long doubling time. To investigate the main reasons for the slow growth and low viability of suspended Vero cells, this study conducted transcriptomic analysis of suspended Vero cells (Vero-XF) and adherent Vero cells (Vero-AD) to screen the differentially expressed genes (DEGs) affecting the growth of suspended cells. In addition, epidermal growth factor (EGF) was supplemented to the culture system to improve the growth of Vero-XF. The results showed that compared with the Vero-AD group, the Vero-XF group had 7 376 significant DEGs. Kyoto encyclopedia of genes and genomes enrichment analysis revealed that the DEGs were mainly enriched in the autophagy and mitophagy pathways. Eleven DEGs were selected and verified by quantitative real-time PCR, which showed up-regulated expression of ATG9B, WIPI2, LAMP2, OPTN, Rab7a, and DEPTOR and down-regulated expression of ATG4D, being consistent with the results of transcriptomic analysis. In addition, the Vero-XF group showed significantly up-regulated expression of ATG101, ATG2A, and STX17 and insignificant change in the expression of NBR1, compared with the Vero-AD group. The protein levels of LC3 and P62 in Vero-XF and Vero-AD were determined by Western blotting, which showed up-regulated expression of LC3Ⅱ/Ⅰ and down-regulated expression of P62 in Vero-XF, indicating a higher level of autophagy. Finally, the exogenous supplementation of EGF at 10, 20, and 30 μg/L in the culture system reduced the autophagy level of Vero-XF by 22.35%, 48.15%, and 71.29%, increased the specific growth rate by 15.48%, 33.33%, and 57.14%, and decreased the apoptosis rate by 2.84%, 15.46%, and 16.23%, respectively. The results of this study preliminarily reveal that the activation of autophagy is one of the reasons for the slow growth of Vero-XF, which provides reference for the subsequent culture of suspended Vero cells.
Animals ; Vero Cells ; Autophagy/genetics* ; Chlorocebus aethiops ; Epidermal Growth Factor/pharmacology* ; Gene Expression Profiling ; Transcriptome ; Cell Survival

Objective To investigate the distribution and antimicrobial resistance profiles of clinically isolated Haemophilus influenzae and Moraxella catarrhalis in hospitals across China from 2015 to 2021,and provide evidence for rational use of antimicrobial agents.Methods Data of H.influenzae and M.catarrhalis strains isolated from 2015 to 2021 in CHINET program were collected for analysis,and antimicrobial susceptibility testing was performed by disc diffusion method or automated systems according to the uniform protocol of CHINET.The results were interpreted according to the CLSI breakpoints in 2022.Beta-lactamases was detected by using nitrocefin disk.Results From 2015 to 2021,a total of 43 642 strains of Haemophilus species were isolated,accounting for 2.91％of the total clinical isolates and 4.07％of Gram-negative bacteria in CHINET program.Among the 40 437 strains of H.influenzae,66.89％were isolated from children and 33.11％were isolated from adults.More than 90％of the H.influenzae strains were isolated from respiratory tract specimens.The prevalence of β-lactamase was 53.79％in H.influenzae strains.The H.influenzae strains isolated from children showed higher resistance rate than the strains isolated from adults.Overall,779 strains of H.influenzae did not produce β-lactamase but were resistant to ampicillin(BLNAR).Beta-lactamase-producing strains showed significantly higher resistance rates to these antimicrobial agents than the β-lactamase-nonproducing strains.Of the 16 191 M.catarrhalis strains,80.06％were isolated from children and 19.94％isolated from adults.M.catarrhalis strains were mostly susceptible to both amoxicillin-clavulanic acid and cefuroxime,evidenced by resistance rate lower than 2.0％.Conclusions The emergence of antibiotic-resistant H.influenzae due to β-lactamase production poses a challenge for clinical anti-infective treatment.Therefore,it is very important to implement antibiotic resistance surveillance for H.influenzae and guide rational antibiotic use.All local clinical microbiology laboratories should actively improve antibiotic susceptibility testing and strengthen antibiotic resistance surveillance for H.influenzae.

Objective To understand the changing composition and antibiotic resistance of bacterial species in the clinical isolates from outpatient and emergency department(hereinafter referred to as outpatients)and inpatient children over time in various hospitals,and to provide laboratory evidence for rational antibiotic use.Methods The data on clinically isolated pathogenic bacteria and antimicrobial susceptibility of isolates from outpatients and inpatient children in the CHINET program from 2015 to 2021 were collected and analyzed.Results A total of 278 471 isolates were isolated from pediatric patients in the CHINET program from 2015 to 2021.About 17.1％of the strains were isolated from outpatients,primarily group A β-hemolytic Streptococcus,Escherichia coli,and Staphylococcus aureus.Most of the strains(82.9％)were isolated from inpatients,mainly SS.aureus,E.coli,and H.influenzae.The prevalence of methicillin-resistant S.aureus(MRSA)in outpatients(24.5％)was lower than that in inpatient children(31.5％).The MRSA isolates from outpatients showed lower resistance rates to the antibiotics tested than the strains isolated from inpatient children.The prevalence of vancomycin-resistant Enterococcus faecalis or E.faecium and penicillin-resistant S.pneumoniae was low in either outpatients or inpatient children.S.pneumoniae,β-hemolytic Streptococcus and S.viridans showed high resistance rates to erythromycin.The prevalence of erythromycin-resistant group A β-hemolytic Streptococcus was higher in outpatients than that in inpatient children.The prevalence of β-lactamase-producing H.influenzae showed an overall upward trend in children,but lower in outpatients(45.1％)than in inpatient children(59.4％).The prevalence of carbapenem-resistant Klebsiella pneumoniae(CRKpn),carbapenem-resistant Pseudomonas aeruginosa(CRPae)and carbapenem-resistant Acinetobacter baumannii(CRAba)was 14％,11.7％,47.8％in outpatients,but 24.2％,20.6％,and 52.8％in inpatient children,respectively.The prevalence of multidrug-resistant E.coli,K.pneumoniae,Proteus mirabilis,P.aeruginosa and A.baumannii strains was lower in outpatients than in inpatient children.The prevalence of fluoroquinolone-resistant E.coli,ESBLs-producing K.pneumoniae,ESBLs-producing P.mirabilis,carbapenem-resistant E.coli(CREco),CRKpn,and CRPae was lower in children in outpatients than in inpatient children,but the prevalence of CRAba in 2021 was higher than in inpatient children.Conclusions The distribution of clinical isolates from children is different between outpatients and inpatients.The prevalence of MRSA,ESBL,and CRO was higher in inpatient children than in outpatients.Antibiotics should be used rationally in clinical practice based on etiological diagnosis and antimicrobial susceptibility test results.Ongoing antimicrobial resistance surveillance and prevention and control of hospital infections are crucial to curbing bacterial resistance.

Objective To investigate the changing antibiotic resistance profiles of E.coli isolated from patients in the 52 hospitals participating in the CHINET program from 2015 to 2021.Methods Antimicrobial susceptibility was tested for clinical isolates of E.coli according to the unified protocol of CHINET program.WHONET 5.6 and SPSS 20.0 software were used for data analysis.Results Atotal of 289 760 nonduplicate clinical strains ofE.coli were isolated from 2015 to 2021,mainly from urine samples(44.7±3.2)％.The proportion of E.coli strains isolated from urine samples was higher in females than in males(59.0％vs 29.5％).The proportion of E.coli strains isolated from respiratory tract and cerebrospinal fluid samples was significantly higher in children than in adults(16.7％vs 7.8％,0.8％vs 0.1％,both P＜0.05).The isolates from internal medicine department accounted for the largest proportion(28.9±2.8)％with an increasing trend over years.Overall,the prevalence of ESBLs-producing E.coli and carbapenem resistant E.coli(CREco)was 55.9％and 1.8％,respectively during the 7-year period.The prevalence of ESBLs-producing E.coli was the highest in tertiary hospitals each year from 2015 to 2021 compared to secondary hospitals.The prevalence of CREco was higher in children's hospitals compared to secondary and tertiary hospitals each year from 2015 to 2021.The prevalence of ESBLs-producing E.coli in tertiary hospitals and children's hospitals and the prevalence of CREco in children's hospitals showed a decreasing trend over the 7-year period.The prevalence of CREco in secondary and tertiary hospitals increased slowly.Antibiotic resistance rates changed slowly from 2015 to 2021.Carbapenem drugs(imipenem,meropenem)were the most active drugs amongβ-lactams against E.coli(resistance rate≤2.1％).The resistance rates of E.coli to β-lactam/β-lactam inhibitor combinations(piperacillin-tazobactam,cefoperazone-sulbactam),aminoglycosides(amikacin),nitrofurantoin and fosfomycin(for urinary isolates only)were all less than 10％.The resistance rate of E.coli strains to antibiotics varied with the level of hospitals and the departments where the strains were isolated,especially for cefazolin and ciprofloxacin,to which the resistance rate of E.coli strains from children in non-ICU departments was significantly lower than that of the strains isolated from other departments(P＜0.05).The E.coli isolates from ICU showed higher resistance rate to most antimicrobial agents tested(excluding tigecycline)than the strains isolated from other departments.The E.coli strains isolated from tertiary hospitals showed higher resistance rates to the antimicrobial agents tested(excluding tigecycline,polymyxin B,cefepime and carbapenems)than the strains from secondary hospitals and children's hospitals.Conclusions E.coli is an important pathogen causing clinical infection.More than half of the clinical isolates produced ESBL.The prevalence of CREco is increasing in secondary and tertiary hospitals over the 7-year period even though the overall prevalence is still low.This is an issue of concern.

Objective:To explore the relationship between serum omentin-1 and 25-hydroxyvitamin D 3 [25(OH)D 3] levels and bone age index (BAI), pelvic ultrasound findings in girls with central precocious puberty (CPP). Methods:This was a cross-sectional observational study. A total of 150 girls with CPP admitted to the Shaanxi Provincial People′s Hospital from October 2021 to October 2023 were selected as the CPP group, and 63 normal girls who underwent physical examination at the child health clinic during the same period were selected as the control group. Serum Omentin-1 and 25(OH)D 3 levels were detected; anteroposterior X-ray and pelvic ultrasound examinations were performed to obtain BAI, ovarian volume, and uterine volume. Pearson correlation analysis was used to evaluate the correlations of Omentin-1 and 25(OH)D 3 with BAI, ovarian volume, and uterine volume. Multivariate logistic regression analysis was used to screen the influencing factors of CPP, and receiver operating characteristic (ROC) curve was drawn to analyze the value of Omentin-1 and 25(OH)D 3 in diagnosing CPP. Results:The serum levels of Omentin-1 and 25(OH)D 3 in the CPP group were lower than those in the control group (all P<0.05); BAI, uterine volume, and ovarian volume in the CPP group were larger than those in the control group, and the number of follicles with diameter ≥4 mm was more than that in the control group (all P<0.05). In girls with CPP, serum Omentin-1 and 25(OH)D 3 levels were negatively correlated with BAI, uterine volume, ovarian volume, and the number of follicles with diameter ≥4 mm (all P<0.05). Multivariate logistic regression analysis showed that maternal menarche age ≤12 years, high BMI, high BAI, and high estradiol were risk factors for CPP (all P<0.05), while high Omentin-1 and high 25(OH)D 3 were protective factors for CPP (all P<0.05). The areas under the curve (AUC) of Omentin-1 and 25(OH)D 3 for diagnosing CPP were 0.799 and 0.808 respectively; the AUC of combined diagnosis was 0.886, which was higher than that of single diagnosis (all P<0.05). Conclusions:Decreased serum Omentin-1 and 25(OH)D 3 levels in girls with CPP are associated with high BAI and increased ovarian and uterine volumes. The combination of Omentin-1 and 25(OH)D 3 has high value in the diagnosis of CPP.

Since the 1980 s, AIDS has been a significant global public health threat. With the "95-95-95" targets for HIV/AIDS prevention and control, the HIV/AIDS "prevention, testing, and treatment" care cascade covering the entire process has become a core strategy. The present study examines combs the connotation and development of the HIV/AIDS care cascade, exploring application throughout the entire process and at each linkage. It analyzes extension value in key populations and HIV co-infected other infectious diseases, identifies obstacles in practice, and proposes strategies to help optimize, innovate, and precisely target HIV/AIDS prevention and control in China.

Objective:To investigate the preventive effect of a subanesthetic dose of esketamine on emergence agitation (EA) in pediatric patients undergoing fracture fixation surgery under general anesthesia.Methods:This study was a randomized, double-blind, placebo-controlled trial. Eighty-two pediatric patients, aged 3-6 yr, with American Society of Anesthesiologists Physical Status classification Ⅰ or Ⅱ, scheduled for elective closed reduction and internal fixation of humeral fractures at the People′s Hospital of Fuyang City from March 2023 to August 2024, were divided into 2 groups ( n=41 each) using simple random sampling: esketamine group (group E) and control group (group C). Group E received an intravenous injection of esketamine at a dose of 0.3 mg/kg before anesthesia induction, while the equal volume of normal saline was given instead in group C. The induction and maintenance of general anesthesia were the same in both groups. The drug was discontinued after operation and the children were transferred to the post-anesthesia care unit. The primary outcome was the occurrence of EA assessed using the Pediatric Anesthesia Emergence Delirium scale. Secondary outcome measures included the acceptance of the facemask during anesthesia induction, the rate of propofol use during the emergence period, the Children′s Hospital of Eastern Ontario Pain Scale scores at awakening and at 2 and 6 h postoperatively, the requirement for rescue analgesia in the emergence period, and the occurrence of adverse reactions. Results:Compared with group C, the incidence of EA [38% (15/39) vs. 15% (6/40)] was significantly decreased, the usage rate of propofol during the emergence period was decreased, the degree of acceptance of the facemask during anesthesia induction was increased, and Children′s Hospital of Eastern Ontario Pain Scale scores at all time points and the rate of rescue analgesia in the emergence period were decreased in group E ( P<0.05). There was no statistically significant difference in the incidence of postoperative adverse reactions between the two groups ( P>0.05). Conclusions:Administering a subanesthetic dose of esketamine before anesthesia induction can reduce the risk of EA in pediatric patients undergoing fracture fixation surgery under general anesthesia and raise the the quality of recovery with a high safety.

Objective This study employs bibliometrics and visualization analysis techniques to systematically review and analyze the current status and evolutionary trends of research on Dysosma versipellis,both domestically and internationally.The discussion encompasses the current state of research on Dysosma versipellis as well as the latest hot topics in the field.Methods The literatures related to Dysosma versipellis and meeting the inclusion criteria were searched through China Knowledge Network(CNKI)and Web of science(WOS)databases,and the core author groups,cooperative relationships among research institutions and key words were analyzed by CiteSpace software.Results Eventually,1793 Chinese documents and 771 English documents were included.The author with the highest number of Chinese articles is Zeng Kang;The authors with the highest number of articles in English are Arora Rajesh;The number of articles published in China generally increases first and then decreases;The Shanghai Institute of Pharmaceutical Industry is the institution that studies Dysosma versipellis the most,and the exchanges and cooperation among institutions in the research field need to be further deepened.The keyword analysis shows that the research content of Dysosma versipellis mainly focuses on anti-tumor application,extraction and synthesis of derivatives.Conclusion At present,the extraction and synthesis of lignans and their related derivatives from Dysosma versipellis and their clinical application in the treatment of condyloma acuminatum and anti-tumor are the research hotspots of Dysosma versipellis.The toxicological mechanisms of damage caused by Dysosma versipellis and its active components,as well as potential therapeutic approaches,are expected to become a focal point of future research.

Objective To summarize the changing prevalence of carbapenem resistance in Enterobacterales based on the data of CHINET Antimicrobial Resistance Surveillance Program from 2015 to 2021 for improving antimicrobial treatment in clinical practice.Methods Antimicrobial susceptibility testing was performed using a commercial automated susceptibility testing system according to the unified CHINET protocol.The results were interpreted according to the breakpoints of the Clinical & Laboratory Standards Institute(CLSI)M100 31st ed in 2021.Results Over the seven-year period(2015-2021),the overall prevalence of carbapenem-resistant Enterobacterales(CRE)was 9.43％(62 342/661 235).The prevalence of CRE strains in Klebsiella pneumoniae,Citrobacter freundii,and Enterobacter cloacae was 22.38％,9.73％,and 8.47％,respectively.The prevalence of CRE strains in Escherichia coli was 1.99％.A few CRE strains were also identified in Salmonella and Shigella.The CRE strains were mainly isolated from respiratory specimens(44.23±2.80)％,followed by blood(20.88±3.40)％and urine(18.40±3.45)％.Intensive care units(ICUs)were the major source of the CRE strains(27.43±5.20)％.CRE strains were resistant to all the β-lactam antibiotics tested and most non-β-lactam antimicrobial agents.The CRE strains were relatively susceptible to tigecycline and polymyxins with low resistance rates.Conclusions The prevalence of CRE strains was increasing from 2015 to 2021.CRE strains were highly resistant to most of the antibacterial drugs used in clinical practice.Clinicians should prescribe antimicrobial agents rationally.Hospitals should strengthen antibiotic stewardship in key clinical settings such as ICUs,and take effective infection control measures to curb CRE outbreak and epidemic in hospitals.

Objective To investigate the correlation between serum levels of macrophage inflammatory protein-1α(MIP-1α),apolipoprotein C1(APOC1),cysteine leukotrienes(CysLTs)and prognosis in children with severe Mycoplasma pneumoniae pneumonia.Methods The children with severe M.pneumoniae pneumonia admitted to the Second Hospital of Handan City from January 2022 to December 2023 were included as case group.The children in case group were assigned to a good prognosis group(n=69)or poor prognosis group(n=24)according to patient outcome.Additionally,93 healthy children who underwent physical examination were included as control group.Enzyme linked immunosorbent assay(ELISA)was applied to determine the serum levels of MIP-1α,APOC1,and CysLTs.Pearson correlation was used to analyze the correlation between serum levels of MIP-1α,APOC1,CysLTs and other biomarkers.Receiver operating characteristic(ROC)curve was plotted to analyze the value of MIP-1α,APOC1,and CysLTs levels for predicting the outcome of children with severe M.pneumoniae pneumonia.Results The serum levels of MIP-1α,APOC1,and CysLTs in case group were higher than those in the control group(P＜0.05).The serum levels of MIP-1α,APOC1,and CysLTs were higher in poor prognosis group compared with good prognosis group(P＜0.05).The levels of MIP-1α,APOC1,and CysLTs in the serum of case group were positively correlated with platelet count,mean platelet volume,white blood cell,neutrophil count,lactate dehydrogenase,erythrocyte sedimentation rate,C-reactive protein,and D-dimer(P＜0.05),and negatively correlated with AT-Ⅲ(P＜0.05).ROC analyses showed that the area under the curve(AUC)of MIP-1α,APOC1,and CysLTs combined was 0.881 in predicting prognosis,significantly higher than MIP-1α(Z=2.096,P=0.036),APOC1(Z=2.236,P=0.025),and CysLTs(Z=2.058,P=0.040)alone,with a sensitivity of 70.80％and specificity of 89.90％.Conclusions The levels of serum MIP-1α,APOC1,and CysLTs are elevated in children with severe M.pneumoniae pneumonia.Serum MIP-1α,APOC1,and CysLTs combined can provide higher value for predicting the prognosis of children with severe M.pneumoniae pneumonia.