BACKGROUND: Temozolomide (TMZ) resistance is a significant challenge in treating glioblastoma (GBM). Collagen remodeling has been shown to be a critical factor for therapy resistance in other cancers. This study aimed to investigate the mechanism of TMZ chemoresistance by GBM cells reprogramming collagens. METHODS: Key extracellular matrix components, including collagens, were examined in paired primary and recurrent GBM samples as well as in TMZ-treated spontaneous and grafted GBM murine models. Human GBM cell lines (U251, TS667) and mouse primary GBM cells were used for in vitro studies. RNA-sequencing analysis, chromatin immunoprecipitation, immunoprecipitation-mass spectrometry, and co-immunoprecipitation assays were conducted to explore the mechanisms involved in collagen accumulation. A series of in vitro and in vivo experiments were designed to assess the role of the collagen regulators prolyl 4-hydroxylase subunit alpha 1 (P4HA1) and yes-associated protein (YAP) in sensitizing GBM cells to TMZ. RESULTS: This study revealed that TMZ exposure significantly elevated collagen type I (COL I) expression in both GBM patients and murine models. Collagen accumulation sustained GBM cell survival under TMZ-induced stress, contributing to enhanced TMZ resistance. Mechanistically, P4HA1 directly binded to and hydroxylated YAP, preventing ubiquitination-mediated YAP degradation. Stabilized YAP robustly drove collagen type I alpha 1 ( COL1A1) transcription, leading to increased collagen deposition. Disruption of the P4HA1-YAP axis effectively reduced COL I deposition, sensitized GBM cells to TMZ, and significantly improved mouse survival. CONCLUSION P4HA1 maintained YAP-mediated COL1A1 transcription, leading to collagen accumulation and promoting chemoresistance in GBM.
Temozolomide ; Humans ; Glioblastoma/drug therapy* ; Animals ; Mice ; Cell Line, Tumor ; Drug Resistance, Neoplasm/genetics* ; YAP-Signaling Proteins ; Hydroxylation ; Dacarbazine/pharmacology* ; Adaptor Proteins, Signal Transducing/metabolism* ; Transcription Factors/metabolism* ; Collagen/biosynthesis* ; Collagen Type I/metabolism* ; Prolyl Hydroxylases/metabolism* ; Antineoplastic Agents, Alkylating/therapeutic use*

Objective:To explore the factors affecting the prognosis of severe pediatric acute respiratory distress syndrome (ARDS) after receiving extracorporeal membrane oxygenation (ECMO) support.Methods:It was a multicenter prospective observational study. A total of 95 children with severe ARDS who were treated with ECMO salvage therapy from January 2018 to December 2022 in 9 pediatric ECMO centers in China were enrolled in the study. The general data, disease severity, organ function, comprehensive treatment and prognosis were recorded, and they were divided into survival group and death group according to the outcome at discharge. T test, chi-square test, multivariate Logistic regression and mixed linear model were used to analyze the relationship among baseline before ECMO treatment, some important indicators (pediatric critical scores, platelet count, albumin, fibrinogen, etc) during ECMO treatment and prognosis. Results:Among the 95 children with severe ARDS who received ECMO, 55 (58%) were males and 40 (42%) were females, aged 36.9 (0.5, 72.0) months. Twelve children (13%) were immunodeficient. Sixty-eight (72%) children were treated with venous artery (VA) mode and 27 (28%) with venous vein (VV) mode. The discharge survival rates of overall, VA, and VV mode children were 51% (48/95), 47% (32/68), and 59% (16/27), respectively. The number of immunodeficient children in the death group was higher, and there were lower pediatric critical scores, platelet count, albumin, fibrinogen and arterial oxygen partial pressure/fraction of inspired oxygen (PaO 2/FiO 2), higher ventilator driving pressure (ΔP), oxygenaion index (OI), and longer ARDS duration before ECMO (all P<0.05). There were no statistically significant differences in other indicators, including age, gender, weight, and ECMO mode among different prognostic groups (all P>0.05). High ΔP, high OI, low P/F, and low albumin were high-risk factors affecting prognosis(all P<0.05). After further grouping, it was found that ΔP≥25 cmH 2O (1 cmH 2O=0.098 kPa), P/F≤67 mmHg (1 mmHg=0.133 kPa) and OI≥35 were the thresholds for predicting poor prognosis ( P<0.05). From 24 h after ECMO, there were significant differences in ΔP, P/F and OI between the dead group and the survival group (all P<0.05), and the differences gradually increased with the ECMO process. The platelet level was significant from 7 days after ECMO ( P<0.05) and gradually expanded. Blood lactate levels showed a significant difference between the 2 groups on before and after ECMO ( P<0.05) and gradually increased from 24 h after ECMO. Conclusions:The risk factors affecting the prognosis of severe ARDS in ECMO include high ΔP, high OI, low P/F and low albumin purification therapy before ECMO. The gradual decrease of ΔP, OI and increase of P/F from 24 h of ECMO predicted a good prognosis, while the gradual increase of lactate after ECMO application showed a poor prognosis.

Objective To analyze the concentration and exposure of vancomycin in children with gram-positive coccal infection,and the corresponding clinical efficacy and safety to support rational use of vancomycin in children.Methods We prospectively collected the clinical and laboratory data of 87 children with gram-positive coccal infection in the Children's Hospital of Fudan University from January 2012 to March 2021.Therapeutic drug monitoring(TDM)was conducted for vancomycin simultaneously,to acquire the data of serum through concentration(Cmin),peak concentration(Cmax),the area under the drug concentration-time curve in a 24-h interval(AUC0-24h)and the ratio ofAUC0-24h to the minimum inhibitory concentration(AUC0-24h/MIC).Results The median(P25,P75)age of the children enrolled in this study was 3.60(1.20,20.00)months.The median dose of vancomycin was 39.23(30.00,46.51)mg/kg.The median serum Cmin was 3.30(1.50,7.10)mg/L.Cmin achieved the target(5-15 mg/L)in 23 cases(26.4％).The median AUC0-24h was 213(174,293)mg·h/L and the median AUC0-24h/MIC was 221(128,349).Adaily dose of above 60 mg/kg in children could achieve the median value of AUC0-24h and AUC0-24h/MIC greater than 400,and the corresponding median age was 28.50(6.85,36.00)months.Multivariate logistic analysis showed a good correlation between Cmin and AUC0-24h(P=0.002).At the end of treatment,the clinical efficacy rate was 85.1％(74/87)and the bacterial eradication rate was 95.4％(83/87).No renal injury occurred during the treatment.Conclusions In this study,the median daily dosage,Cmin and A UC0-24h/MIC of vancomycin were below the recommended range at home and abroad.However,good clinical and microbiological efficacy were achieved in children at low vancomycin exposure.The probability of target attainment(PTA)for A UC0-24h and AUC0-24h/MIC ≥400 increased when the daily dose of vancomycin was＞60 mg/kg(corresponding to the median age of 28.50 months)or Cmin ≥ 5 mg/L.

Objective:To investigate the driving effect of prostate cancer associated transcript 4 (PCAT4) on the up-regulation of upregulator of cell proliferation (URGCP) expression in breast cancer progression through sponging miR-508-5p.Methods:The microarray data of lncRNA and miRNA with differential expression in breast cancer tissue were analyzed by Cancer Genome Atlas. The expression of PCAT4 in breast cancer was evaluated by real-time quantitative polymerase chain reaction (RT-qPCR). Cell proliferation was measured by MTT and colony formation, cell apoptosis was analyzed by TUNEL, and cell migration and invasion were analyzed by Transwell. The correlation between PCAT4 and miR-508-5p, and miR-508-5p and URGCP was analyzed by RNA pull-down and double luciferase assay. Tumor xenograft studies were performed to analyze the correlation between PCAT4/miR-508-5p/URGCP axis and breast cancer cell growth in vivo. Measurement data were expressed as mean ± standard deviation ( ± s). T-test was used for comparison between two groups, and one-way analysis of variance was used for comparison between multiple groups. The correlation between PCAT4 and URGCP and miR-508-5p expression was evaluated by Pearson correlation analysis. Results:The expression level of PCAT4 was up-regulated in breast cancer tissues and cells. Knockout of PCAT4 inhibited cell proliferation and metastasis and promoted cell apoptosis. miR-508-5p was the target of PCAT4 and was negatively correlated with PCAT4. Overexpression of miR-508-5p in breast cancer can inhibit cell growth, migration and invasion, and promote cell apoptosis. URGCP is the target of miR-508-5p and induces progression of breast cancer. Tumor xenograft studies showed that PCAT4 drives breast cancer progression by affecting miR-508-5p/URGCP.Conclusion:The expression of PCAT4 is up-regulated in breast cancer tissues and cells, and PCAT4 can act as a molecular sponge of miR-508-5p, and significantly promote breast cancer progression by activating URGCP protein expression.

Objective:To understand the current situations and existing problems of pediatric emergency in Shanghai city and provide a basis for the construction and management of pediatric emergency.Methods:The questionnaire survey was used to investigate the current situations of pediatric emergency department in secondary and tertiary public hospitals in Shanghai city.Results:A total of 28 questionnaires were delivered, and 28 were responded.Six of the 28 hospitals had no administratively independent pediatric emergency or no separate pediatric emergency area.Of the 22 hospitals, each had an average of 9.7 professional emergency pediatricians, with the exception of one secondary hospital that lacked professional emergency pediatricians.Professional emergency pediatricians accounted for an average of 70.0% of all pediatricians.Of the 22 hospitals, 18(81.8%) were open 24 hours a day.Fourteen(63.6%) had independent triage.Eight children′s emergency rooms(36.4%) were shared with adults.Among the 22 hospitals, 20 hospitals filled in the annual total number of pediatric emergency visits, and the total number of pediatric emergency visits ranged from 2 791 to 467 428, with an average of 93 966.65.Conclusion:There are still some problems in the development of pediatric emergency department in secondary and tertiary public hospitals in Shanghai city, such as insufficient human resources, substandard personnel training, incomplete equipment and lack of critical first-aid techniques.Therefore, the construction and management of pediatric emergency department still need to be improved.

Infectious diseases are clinically very common, among which bacterial infections are the most common ones.Therefore, antibiotics have become ones of the most widely used drugs in clinical practice.While saving lives of many infected patients, antibiotics can also lead to adverse reactions, which can cause damage to organs function in severe cases and even life-threatening.Common adverse reactions include allergic reactions and various organs damage.In this review, we aimed to focus on the evaluation of common adverse reactions and organs damage caused by antibiotics, in order to improve the prevention and treatment of adverse reactions and promote rational use of antibiotics.

Objective:To investigate the effect of activation of microglia in prefrontal cortex on long-term spatial memory in post-stroke depression mice.Methods:Forty-eight male C57BL/6 mice were divided into sham operation group, stroke group, post-stroke depression group and depression group according to the random number table method with 12 in each group, and 36 mice were divided into solvent group, enrofloxacin group and minocycline group according to the random number table method with 12 in each group.Middle cerebral artery occlusion (MCAO) was use to establish the stroke model, and forced swimming was used to establish the depression model.The post-stroke depression model mice were received MCAO first and then received forced swimming on the 4th day after stroke to establish the model.Mice in enrofloxacin group and minocycline group were treated with enrofloxacin and minocycline injection once a clay for 14 days from the 5th day after stroke, respectively.Forced swimming test and sugar water preference test were used to evaluate the depression of mice in each group, Morris water maze test was used to detect the spatial memory function of mice in each group, and Nissl staining and immunofluorescence staining were used to detect the neuronal function and the number and type of microglia activation.The expression of inflammatory cytokines IL-6 and IL-1β were detected by Western blot.GraphPad Prism 8.0.1 statistical software was used for statistical analysis.The single factor variance analysis was used to compare the difference among multiple groups, and pairwise comparison was performed with SNK- q test. Results:(1) There were statistically significant differences in depression, learning and memory, neuron damage, activation of microglia, inflammatory factors and other indicators in sham operation group, stroke group, post-stroke depression group and depression group ( F=43.58-255.70, all P<0.05). Compared with stroke group, post-stroke depression group had longer floating immobility time ((222.70±29.12) s, (79.25±46.78) s, P<0.05), the preference rate of sugar water was significantly lower ( (49.44±6.19) %, (84.49±4.73) %, P<0.05), and the average value of platform approach after correction was higher((125.00±9.95) mm, (96.79±12.57) mm, P<0.05), Nissl bodies expression was lower ((53.50±15.78) cells /mm 2, (85.67±17.52) cells /mm 2, P<0.05), NeuN positive expression rate was lower ((29.78±3.70) %, (45.73±4.51) %, P<0.05), the percent of M1 microglia expression was significantly higher ((75.55±8.84) %, (58.19±5.69) %, P<0.05), the percent of M2 microglia expression was lower ((43.46±5.11)%, (57.14±5.40)%, P<0.05), and the expression levels of IL-6 ((1.14±0.03), (0.94±0.05), P<0.05) and IL-1β((1.17±0.03), (0.56±0.04), P<0.05) were significantly higher.(2) Depression, learning and memory, neuron injury, activation of microglia, inflammatory factors and other indicators of mice in solvent group, enrofloxacin group and minocycline group were significantly different ( F=7.13-94.35, all P<0.05). Compared with enrofloxacin group, mice in minocycline group had shorter floating immobility time ((169.30±13.04) s, (224.30±22.60) s, P<0.05) and higher sugar water preference rate ((62.81±7.75) %, (47.71±8.11) %, P<0.05), the mean value of platform approach estimation after water maze correction was lower ((97.66±14.56) mm, (120.20±12.08) mm, P<0.05), and the expression level of Nissl bodies was higher ((80.17±10.55) cells /mm 2, (52.00±8.94) cells /mm 2, P<0.05), NeuN expression rate was high ((45.04±3.62) %, (28.88±4.50) %, P<0.05), Iba-1 expression was lower ((97.33±10.67) cells/mm 2, (112.50±6.54)cells/mm 2, P<0.05), the percent of M1 microglia expression was lower ((54.43±5.22) %, (73.82±6.88) %, P<0.05), and the percent of M2 microglia expression was significantly higher ((51.86±6.22) %, (36.30±5.72) %, P<0.05). The expression levels of IL-6 ((0.75±0.06), (1.21±0.07), P<0.05) and IL-1β ((0.61±0.06) (1.09±0.09), P<0.05) were lower. Conclusion:The long-term spatial memory impairment of post-stroke depression mice is aggravated, which is related to the neuron damage caused by increased activation of M1 microglia in PFC area.Inhibition of M1 microglia by minocycline can effectively improve the spatial memory ability of mice.

Objective:To evaluate the effectiveness of integrated pediatric training course of extracorporeal membrane oxygenation(ECMO)based on simulation teaching.Methods:The instructors of ECMO team of Children′s Hospital of Fudan University developed the teaching curriculum.Team members as a unit were recruited to study simulation-based courses, whose theoretical knowledge, skills and teamwork were assessed, and feedback from the trainees were gathered.Results:Since May 2018, 22 teams of pediatric specialized hospitals or general hospitals have taken part in our ECMO simulation, with totally 132 participants, including 45 ICU doctors(34.1%), 60 ICU nurses(45.5%), 23 thoracic surgeons(17.4%) and four anesthesiologists(3.0%). Twelve of them(9.1%)had a little ECMO experience.After training, the trainees had a high evaluation on each part of the course, with average score more than four, of which the theoretical score was lowest.Their self-evaluation on the theory, skills, teamwork and confidence in the implementation of ECMO had been significantly improved.Most(80.3%)of the trainees were confident to carry out ECMO in their local hospitals.The trainees′ baseline score of theoretical knowledge was 55.2±7.6, and increased to 67.1±7.3 after training( P<0.001). The average teamwork score of the 22 teams was 70.2±8.2, and the qualified rate was up to 86.4%.The most prominent skill problems in operation assessment were no albumin and blood priming(90.9%), non-skill problems were extremely anxious during the overall assessment(84.8%)of the participants, poor team work(74.2%), poor leadership(68.2%). For the feedback of the whole course, 97.0% and 94.7% of the trainees thought that integrated training and simulation teaching were the highlights of the course.At present, 13(59.1%)units have successfully developed ECMO technology after the course, and the number of children treated has been up to 83, with a survival rate of discharge of 54.2%.However, it was found that the skill level of some participants decreased about 2 weeks after the course. Conclusion:As a new attempt of ECMO teaching, this curriculum has achieved certain efficiency in both teaching and clinical practice.At the same time, our study also plays a role in promoting the development of ECMO in pediatrics.

Objective:To explore the value of body temperature, pediatric clinical illness score(PCIS), white blood cell count (WBC), plasma C-reactive protein (CRP), procalcitonin (PCT) and pro-adrenomedullin (pro-ADM) in predicting nosocomial infection in PICU.Methods:From June 2016 to March 2017, the critically ill children in PICU of Children′s Hospital of Fudan University were selected and divided into nosocomial infection group and non nosocomial infection group according to the diagnostic criteria of nosocomial infection.The body temperature, PCIS, WBC, CRP, PCT and pro-ADM were recorded at 4 hours (T1), (48±1) hours (T2), (120±1) hours (T3) and (192±1) hours (T4) after admission, and their predictive value of each index, which was the closest time point (Th) to nosocomial infection was analyzed.Receiver-operating characteristic (ROC) curves were performed to calculate the areas under the curves (AUC), sensitivity and specificity, and multivariate Logistic regression analysis was used to study the risk factors of nosocomial infection.Results:A total of 85 cases were included, including 27 cases in nosocomial infection group and 58 cases in non nosocomial infection group.There was no significant difference in age, weight, body temperature, WBC, PCT, pro-ADM, primary disease and invasive operation between two groups (all P>0.05). There were significant differences in gender, PCIS, CRP, intubation rate and central venous catheterization rate ( P<0.05), when patients were admitted to PICU.At Th, the differences of body temperature, PCIS, CRP, PCT and pro-ADM between two groups were statistically significant ( P<0.05), as well as the AUC were 0.787, 0.755, 0.709, 0.704 and 0.809, respectively, as well as the best cut-off values for predicting nosocomial infection were 38.0 ℃, 87 points, 14.5 mg/L, 0.28 ng/mL and 0.67 nmol/L, respectively.There was no significant difference regarding WBC between two groups ( P>0.05). PCIS may be an independent risk factor for nosocomial infection( OR=0.978, 95% CI 95.9-99.9, P<0.05). Conclusion:Pro-ADM has high sensitivity and specificity in predicting nosocomial infection, and PCIS is an independent risk factor for nosocomial infection.