Objective:To investigate the effect of glioma cell-derived apoptotic extracellular vesicles(apoEVs)on glioma tumorigen-esis and temozolomide(TMZ)resistance and its mechanism.Methods:The extracted apoEVs were characterized using nanoparticle tracking analysis and transmission electron microscopy,and Western blot,flow cytometry,CCK-8 assay,colony formation assay,and Transwell assay were used to investigate the effect of apoEVs on cell proliferation,migration,invasion,and apoptosis.Results:ApoEVs promoted TMZ resistance in glioma cells,significantly increased the half-maximal inhibitory concentration of TMZ(t=9.326,P=0.001),and inhibited cell apoptosis,and this effect could be reversed by the exosome inhibitor GW4869.ApoEVs also promoted the migration and invasion of glioma cells,increased the expression of vimentin and Twist proteins(t=8.762,P=0.002;t=7.941,P=0.004),and inhibited the expression of cleaved caspase-3(t=9.217,P=0.002).Further studies showed that apoEVs affected TMZ resistance of glioma cells by regulating the LncRNA-XIST/miR-29c/O6-methylguanine-DNA methyltransferase(MGMT)axis.Silencing of LncRNA-XIST could reduce the expression of MGMT and increase the expression of miR-29c,thereby enhancing the sensitivity to TMZ and inhibiting cell migration and invasion.Conclusion:Glioma cell-derived apoEVs promote the malignant progression and temo-zolomide resistance of glioma by delivering LncRNA-XIST to regulate the miR-29c/MGMT axis.

Objective To explore the efficacy and safety of doxycycline in the treatment of Mycoplasmal pneumoniae pneumonia(MPP)in children under 8 years old.Methods A retrospective selection was made of 258 children with MPP under 8 years old who were admitted to Women and Children's Hospital of Ningbo University from July 2023 to June 2024 as the research subjects.The children were divided into the azithromycin group(n=156)and the doxycycline group(n=102)according to the different drugs used.The clinical indicators,inflammation-related indicators,liver function indicators before and after treatment and adverse reactions of two groups of children were compared.Results After treatment,the defervescence time,cough relief time,pneumonia image absorption time and hospital stay in doxycycline group were shorter than those in azithromycin group(P＜0.05),the number of patients who needed to use steroids and bronchoscope intervention,and the number of patients with total hospitalization costs more than 10 000 yuan were less than those in azithromycin group(P＜0.05).The C-reactive protein(CRP),D-dimer(DD),serum ferritin(SF)and lactate dehydrogenase(LDH)in both groups of children were significantly lower than those before treatment in this group(P＜0.05),and the reduction amplitzes of CRP,DD,SF and LDH in doxycycline group were significantly greater than those in azithromycin group(P＜0.05).There were no significant changes in alanine aminotransferase,aspartate aminotransferase and prealbumin between two groups of children before and after treatment(P＞0.05).The adverse reactions of two groups of children were mainly gastrointestinal reactions,and there was no statistically significant difference in the incidence rate(x2=0.254,P=0.204).Conclusion Doxycycline in treatment of MPP in children under 8 years old can effectively relieve clinical symptoms,reduce inflammatory responses,decrease hormone and bronchoscopy interventions,shorten hospital stays and reduce hospitalization costs,and has good safety.

To investigate the epidemiological characteristics of co-infection involving macrolide-resistant Mycoplasma pneumoniae (MRMP) and non-bacterial pathogens in hospitalized children at Women and Children′s Hospital of Ningbo University from 2021 to 2024, a retrospective cross-sectional study was conducted. Throat swabs were collected from children admitted for respiratory tract infections between January 2021 and December 2024. A total of 22 311 children aged 0-18 years old were included, including 12 021 males and 10 290 females. The median age was 5.00 years and the mean age was (5.02±3.25) years. Real-time fluorescence quantitative polymerase chain reaction (PCR) and multiplex PCR were employed to determine the MP infection rate, the prevalence of MRMP, and the co-infection rates with non-bacterial pathogens across different years. Patients with MRMP were categorized into two groups based on the presence or absence of co-infections: simple infection group and mixed infection group. The differences in age, gender, and onset time between these groups were analyzed, along with the distribution of pathogens in the mixed infection group. Chi-square tests were used for intergroup comparisons. The results showed that the overall positive rate of MP among the 22 311 children was 39.42% (8 794/22 311), with a detection rate of MP drug resistance gene mutations at 70.42% (6 193/8 794). The co-infection rate of MRMP was 24.29% (1 504/6 193). Statistically significant differences were observed in the MP positive rate and the detection rate of drug resistance gene mutations from 2021 to 2024 ( χ2=1 674.420, P<0.05; χ2=67.733, P<0.05), with peak values in 2023 (50.87% and 73.83%, respectively). Among the annual co-infections, the highest rate was in 2024 (28.72%), while the lowest was in 2022 (7.30%). In the age distribution, the rate of mixed infections decreased with increasing age ( χ2=84.742, P<0.05). Seasonally, the infection rates in winter 2023 and the spring, summer, and autumn of 2024 were significantly higher than those in 2022 ( χ2=24.243, 13.101, 26.181, and 10.687, respectively; all P<0.05), with the lowest mixed infection rate observed in winter 2022 over the four-year period. Over the four years, the highest positive rate for rhinovirus was consistently observed in MRMP co-infections each year, particularly in the 3-to 6-year-old age group. The positive rate for mixed infections involving respiratory syncytial virus, human bocavirus, and parainfluenza virus was notably higher in the 0- to 3-year-old age group. Following the removal of non-pharmaceutical interventions (NPIs) in December 2022, the co-infection rate of other respiratory viruses and MRMP increased in Ningbo City. In conclusion, the positive rate of MRMP has shown an upward trend from 2021 to 2023. Post-NPI removal, MRMP mixed infections have become more prevalent in 2023 and 2024, predominantly affecting children aged 0-6 years, with rhinovirus being the most common co-pathogen.

To investigate the epidemiological characteristics of co-infection involving macrolide-resistant Mycoplasma pneumoniae (MRMP) and non-bacterial pathogens in hospitalized children at Women and Children′s Hospital of Ningbo University from 2021 to 2024, a retrospective cross-sectional study was conducted. Throat swabs were collected from children admitted for respiratory tract infections between January 2021 and December 2024. A total of 22 311 children aged 0-18 years old were included, including 12 021 males and 10 290 females. The median age was 5.00 years and the mean age was (5.02±3.25) years. Real-time fluorescence quantitative polymerase chain reaction (PCR) and multiplex PCR were employed to determine the MP infection rate, the prevalence of MRMP, and the co-infection rates with non-bacterial pathogens across different years. Patients with MRMP were categorized into two groups based on the presence or absence of co-infections: simple infection group and mixed infection group. The differences in age, gender, and onset time between these groups were analyzed, along with the distribution of pathogens in the mixed infection group. Chi-square tests were used for intergroup comparisons. The results showed that the overall positive rate of MP among the 22 311 children was 39.42% (8 794/22 311), with a detection rate of MP drug resistance gene mutations at 70.42% (6 193/8 794). The co-infection rate of MRMP was 24.29% (1 504/6 193). Statistically significant differences were observed in the MP positive rate and the detection rate of drug resistance gene mutations from 2021 to 2024 ( χ2=1 674.420, P<0.05; χ2=67.733, P<0.05), with peak values in 2023 (50.87% and 73.83%, respectively). Among the annual co-infections, the highest rate was in 2024 (28.72%), while the lowest was in 2022 (7.30%). In the age distribution, the rate of mixed infections decreased with increasing age ( χ2=84.742, P<0.05). Seasonally, the infection rates in winter 2023 and the spring, summer, and autumn of 2024 were significantly higher than those in 2022 ( χ2=24.243, 13.101, 26.181, and 10.687, respectively; all P<0.05), with the lowest mixed infection rate observed in winter 2022 over the four-year period. Over the four years, the highest positive rate for rhinovirus was consistently observed in MRMP co-infections each year, particularly in the 3-to 6-year-old age group. The positive rate for mixed infections involving respiratory syncytial virus, human bocavirus, and parainfluenza virus was notably higher in the 0- to 3-year-old age group. Following the removal of non-pharmaceutical interventions (NPIs) in December 2022, the co-infection rate of other respiratory viruses and MRMP increased in Ningbo City. In conclusion, the positive rate of MRMP has shown an upward trend from 2021 to 2023. Post-NPI removal, MRMP mixed infections have become more prevalent in 2023 and 2024, predominantly affecting children aged 0-6 years, with rhinovirus being the most common co-pathogen.

Objective To explore the efficacy and safety of doxycycline in the treatment of Mycoplasmal pneumoniae pneumonia(MPP)in children under 8 years old.Methods A retrospective selection was made of 258 children with MPP under 8 years old who were admitted to Women and Children's Hospital of Ningbo University from July 2023 to June 2024 as the research subjects.The children were divided into the azithromycin group(n=156)and the doxycycline group(n=102)according to the different drugs used.The clinical indicators,inflammation-related indicators,liver function indicators before and after treatment and adverse reactions of two groups of children were compared.Results After treatment,the defervescence time,cough relief time,pneumonia image absorption time and hospital stay in doxycycline group were shorter than those in azithromycin group(P＜0.05),the number of patients who needed to use steroids and bronchoscope intervention,and the number of patients with total hospitalization costs more than 10 000 yuan were less than those in azithromycin group(P＜0.05).The C-reactive protein(CRP),D-dimer(DD),serum ferritin(SF)and lactate dehydrogenase(LDH)in both groups of children were significantly lower than those before treatment in this group(P＜0.05),and the reduction amplitzes of CRP,DD,SF and LDH in doxycycline group were significantly greater than those in azithromycin group(P＜0.05).There were no significant changes in alanine aminotransferase,aspartate aminotransferase and prealbumin between two groups of children before and after treatment(P＞0.05).The adverse reactions of two groups of children were mainly gastrointestinal reactions,and there was no statistically significant difference in the incidence rate(x2=0.254,P=0.204).Conclusion Doxycycline in treatment of MPP in children under 8 years old can effectively relieve clinical symptoms,reduce inflammatory responses,decrease hormone and bronchoscopy interventions,shorten hospital stays and reduce hospitalization costs,and has good safety.

OBJECTIVE To investigate the efficacy and safety of doxycycline in the treatment of macrolide-unresponsive Mycoplasma pneumoniae pneumonia(MUMPP)in children under 8 years of age.METHODS The medical records of children with MUMPP admitted to the Women and Children's Hospital Affiliated to Ningbo University were collected from January 1st,2023 to December 31st,2023.They were divided into doxycycline group(44 cases),doxycycline combined with methylprednisolone group(35 cases),and azithromycin combined with methylprednisolone group(35 cases)according to the treatment methods.Doxycycline group was given Doxycycline hyclate enteric-coated capsules;doxycycline combined with methylprednisolone group was given Doxycycline hyclate enteric-coated capsules and Methylprednisolone sodium succinate for injection;azithromycin combined with methylprednisolone group was given Azithromycin for injection and Methylprednisolone sodium succinate for injection.Treatment courses of 3 groups lasted for 10 d.The fever reduction rate,the time of fever reduction and improvement rate of lung infection absorption were compared among the three groups.The occurrence of adverse drug reactions was recorded during their hospitalization and followed up within 5 months after discharge.RESULTS The fever reduction rats 48,72 h after treatment and improvement rate of lung infection absorption in doxycycline group and doxycycline combined with methylprednisolone group were significantly higher than azithromycin combined with methylprednisolone group;the time of fever reduction was significantly shorter than azithromycin combined with methylprednisolone group(P＜0.05);there was no statistical significance in the difference between the doxycycline group and the doxycycline combined with methylprednisolone group(P＞0.05).There was no statistical significance in the incidence of rash,vomiting,abdominal pain,diarrhea,and elevated transaminases among the three groups during hospitalization and within 5 months after discharge(P＞0.05).None of the children treated with doxycycline suffered from tooth discoloration or enamel hypoplasia.CONCLUSIONS Doxycycline has good efficacy and safety in therapy of MUMPP in children under 8 years of age;adjunctive coadministration of low-dose glucocorticoids does not necessarily result in significant additional efficacy.

Objective:To explore potential predictors of refractory Mycoplasma pneumoniae pneumonia (RMPP) in early stage. Methods:The prospective multicenter study was conducted in Zhejiang, China from May 1 st, 2019 to January 31 st, 2020. A total of 1 428 patients with fever >48 hours to <120 hours were studied. Their clinical data and oral pharyngeal swab samples were collected; Mycoplasma pneumoniae DNA in pharyngeal swab specimens was detected. Patients with positive Mycoplasma pneumoniae DNA results underwent a series of tests, including chest X-ray, complete blood count, C-reactive protein, lactate dehydrogenase (LDH), and procalcitonin. According to the occurrence of RMPP, the patients were divided into two groups, RMPP group and general Mycoplasma pneumoniae pneumonia (GMPP) group. Measurement data between the 2 groups were compared using Mann-Whitney U test. Logistic regression analyses were used to examine the associations between clinical data and RMPP. Receiver operating characteristic (ROC) curves were used to analyse the power of the markers for predicting RMPP. Results:A total of 1 428 patients finished the study, with 801 boys and 627 girls, aged 4.3 (2.7, 6.3) years. Mycoplasma pneumoniae DNA was positive in 534 cases (37.4%), of whom 446 cases (83.5%) were diagnosed with Mycoplasma pneumoniae pneumonia, including 251 boys and 195 girls, aged 5.2 (3.3, 6.9) years. Macrolides-resistant variation was positive in 410 cases (91.9%). Fifty-five cases were with RMPP, 391 cases with GMPP. The peak body temperature before the first visit and LDH levels in RMPP patients were higher than that in GMPP patients (39.6 (39.1, 40.0) vs. 39.2 (38.9, 39.7) ℃, 333 (279, 392) vs. 311 (259, 359) U/L, both P<0.05). Logistic regression showed the prediction probability π=exp (-29.7+0.667×Peak body temperature (℃)+0.004×LDH (U/L))/(1+exp (-29.7+0.667×Peak body temperature (℃)+0.004 × LDH (U/L))), the cut-off value to predict RMPP was 0.12, with a consensus of probability forecast of 0.89, sensitivity of 0.89, and specificity of 0.67; and the area under ROC curve was 0.682 (95% CI 0.593-0.771, P<0.01). Conclusion:In MPP patients with fever over 48 to <120 hours, a prediction probability π of RMPP can be calculated based on the peak body temperature and LDH level before the first visit, which can facilitate early identification of RMPP.

Objective:To investigate the pathogenic spectrum of children with severe infection by metagenomic next generation sequencing (mNGS).Methods:This study was a cross-sectional study. We collected 212 cases of severely infected pediatric patients admitted to the Intensive Care Unit (ICU) of the Women and Children′s Hospital of Ningbo University from January 2022 to June 2023, and performed metagenomic next-generation sequencing (mNGS) on 249 samples to analyze the pathogenic distribution characteristics.Results:Among the 249 samples of 212 children, the positive detection rate was 49.80% (124/249), including 14 cases of mixed infections, accounting for 6.60% (14/212). According to the mNGS technology, the pathogen distribution of severely infected children showed that the most common Gram-positive bacteria were Staphylococcus aureus (3.61%, 9/249), Streptococcus pneumoniae (2.81%, 7/249), and Staphylococcus epidermidis (2.41%, 6/249); the most common Gram-negative bacteria were Klebsiella aerogenes (2.41%, 6/249), Klebsiella pneumoniae (2.41%, 6/249), and Haemophilus parainfluenzae (2.01%, 5/249). The most common fungus was Candida parapsilosis (2.01%, 5/249). The most common virus was Human Cytomegalovirus (HCMV) (6.02%, 15/249), Human Herpesvirus 1 (HHV-1) (1.61%, 4/249), and Epstein-Barr virus (EBV) (1.61%, 4/249). The most common atypical pathogen was Mycoplasma pneumoniae (3.21%, 8/249). Conclusions:This study explored the pathogen spectrum in severely infected pediatric patients through mNGS, contributing to the diagnosis of mixed infections or infections caused by uncommon or rare pathogens, which enables rapid and efficient identification of pathogens.

OBJECTIVE: To explore the influencing factors of perioperative blood transfusion in the treatment of elderly femoral intertrochanteric fractures with proximal femoral nail antirotation(PFNA). METHODS: The clinical data of 109 elderly patients with intertrochanteric fractures who received PFNA treatment from July 2018 to January 2020 were retrospectively analyzed. Both pelvic hip X-rays and CT plain scans were performed before surgery. All patients were diagnosed by X-ray and CT plain scan of pelvis and hip before operation. Through the statistical analysis of the basic data of patients before and during operation, the risk factors of perioperative blood transfusion were explored. RESULTS: Logistic regression analysis showed that age ( CONCLUSION Age, fracture type, diabetes history, and preoperative hemoglobin are independent risk factors for perioperative blood transfusion in the treatment of elderly intertrochanteric fractures with PFNA. The older the patient, the history of diabetes, the more unstable the fracture, and the lower preoperative hemoglobin, the more likely it is to require a blood transfusion, which may provide a reference for clinical perioperative blood transfusion decisions.
Aged ; Blood Transfusion ; Bone Nails ; Fracture Fixation, Intramedullary/adverse effects* ; Hip Fractures/surgery* ; Humans ; Retrospective Studies ; Risk Factors ; Treatment Outcome

Objective To generate HLA - A * 0201 and A * 2403 restricted HBcAg - specific cytotoxic T lymphocyte (CTL) clones. Methods Peripheral blood mononuclear cells (PBMCs) were derived from a HLA - A * 0201/2403 - positive patient with chron- ic hepatitis B . PBMCs were stimulated respectively using two synthetic peptides( HBc18 ～ 27 and HBc 117 ～ 125) , and epitope -specific CTL clones were generated by limiting dilution technique with PHA alone or combined with synthetic peptides. The cell clones were then characterized by IF staining,FCM and LDH release. Results After 2 weeks of in vitro stimulating PBMCs, specific CTL lines were established. 29 T clones were generated from those CTL lines using HBc18 ～27 stimulating. Among the 29 clones ,28 clones belonged to CD8~+ T cells and all displayed cytolytic activity. 12 CD8 +T clones were generated from those CTL lines using HBc117 ～ 125 stimulating. Specific cytotoxic activity was observed in 9 of those clones. The other three displayed less cytolytic activity. In the process of cloning, PHA was used alone or combined with synthetic peptides,and the achievement showed a rate of 15.62% and 14.58% . Conclusion HBc18 ～ 27 and HBc117 ～ 125 are capable of activating CD8~+ T cells in PBMCs of HLA - A * 0201 /2403 patient with chronic hepatitis B. In the process of CD8~+T cloning, synthetic peptides could not increase the rate of successful cloning.