BACKGROUND: P16 inactivation is frequently accompanied by telomerase reverse transcriptase ( TERT ) amplification in human cancer genomes. P16 inactivation by DNA methylation often occurs automatically during immortalization of normal cells by TERT . However, direct evidence remains to be obtained to support the causal effect of epigenetic changes, such as P16 methylation, on cancer development. This study aimed to provide experimental evidence that P16 methylation directly drives cancer development. METHODS: A zinc finger protein-based P16 -specific DNA methyltransferase (P16-Dnmt) vector containing a "Tet-On" switch was used to induce extensive methylation of P16 CpG islands in normal human fibroblast CCD-18Co cells. Battery assays were used to evaluate cell immortalization and transformation throughout their lifespan. Cell subcloning and DNA barcoding were used to track the diversity of cell evolution. RESULTS: Leaking P16-Dnmt expression (without doxycycline-induction) could specifically inactivate P16 expression by DNA methylation. P16 methylation only promoted proliferation and prolonged lifespan but did not induce immortalization of CCD-18Co cells. Notably, cell immortalization, loss of contact inhibition, and anchorage-independent growth were always prevalent in P16-Dnmt&TERT cells, indicating cell transformation. In contrast, almost all TERT cells died in the replicative crisis. Only a few TERT cells recovered from the crisis, in which spontaneous P16 inactivation by DNA methylation occurred. Furthermore, the subclone formation capacity of P16-Dnmt&TERT cells was two-fold that of TERT cells. DNA barcoding analysis showed that the diversity of the P16-Dnmt&TERT cell population was much greater than that of the TERT cell population. CONCLUSION P16 methylation drives TERT -mediated immortalization and transformation of normal human cells that may contribute to cancer development.
Humans ; Telomerase/genetics* ; DNA Methylation/physiology* ; Fibroblasts/cytology* ; Cyclin-Dependent Kinase Inhibitor p16/metabolism* ; Cell Line ; Cell Transformation, Neoplastic/genetics*

Objective To investigate the current status of nurses'knowledge,attitude and practice regarding sleep management of critically ill children in pediatric ICU,and to analyze its impact factors.Methods A self-designed questionnaire on general information and a questionnaire on knowledge and practical behaviors of pediatric ICU nurses on child's sleep management were used.In March 2023,902 pediatric ICU nurses from 24 hospitals in China were surveyed using a convenient sampling method,and the impact factors were analyzed using multiple stepwise linear regression.Results 893 valid questionnaires were collected and the recovery rate of valid questionnaires was 99.00％.Nurses in pediatric ICU scored(33.71±7.76)in knowledge dimension,(37.38±4.86)in attitude dimension and(80.60±16.78)in practice dimension,with a total score of(151.78±24.27).The scores of knowledge and attitude,knowledge and practice,attitude and practice are all positively correlated(r=0.393,P＜0.001;r=0.495,P＜0.001;r=0.320,P＜0.001).The results of multiple stepwise linear regression analysis showed that gender,region,whether they had received sleep management training were the influencing factors of pediatric ICU nurses'total score of knowledge,attitude and practice towards children's sleep management(P＜0.05).Conclusion Nurses in pediatric ICU are positive about sleep management for critically ill children,but their knowledge and practice levels need to improve.Nursing managers should strengthen the theoretical knowledge and practical behavioral training of pediatric ICU nurses on child sleep management,develop scientific sleep management plans,and guide nurses to make reasonable evaluation and interventions to improve children's sleep quality.

Objective:To explore the association between self-control and the co-occurrence of depres-sive symptoms and overweight or obesity from adolescence to early adulthood in the Chinese population,and to provide a scientific basis for personalized interventions targeting individuals with different risks in the future.Methods:From a prospective cohort study that lasted for 10 years:The China family panel studies(CFPS),a total of 608 children and adolescents meeting the following inclusion and exclusion criteria were included as study subjects:(1)Aged 10 to 19 years,at normal weight according to Chinese standards,and without depressive symptom in 2010;(2)Had self-control scores,and with at least two measurements of depressive symptoms and body mass index(BMI)between 2010 and 2020;(3)The only one or the youngest child and adolescent from each family.The co-occurrence of depressive symp-toms and overweight or obesity was defined in three ways:Both of the average level of standardized scores of depressive symptoms and BMI Z-scores across multiple measurements over time were at a high level,or both of the trajectories of depressive symptoms and BMI over time based on the latent classification trajec-tory model(LCTM)belonging to the"risk-type",or individuals had depressive symptoms and over-weight/obesity at the last follow-up survey.The multinomial Logistic regression model was used to examine the association between standardized scores of self-control and the co-occurrence of depressive symptoms and overweight or obesity.Results:The score of self-control was associated with the co-occur-rence of depressive symptoms and overweight or obesity when using healthy individuals as the reference group after adjusting for age(years),gender(male/female),area(urban/rural),weekly physical ac-tivity duration(high/low),parental education level(college or above/high school or below),parental weight status(overweight or obese or not),and parental depressive symptoms(with depressive symptoms or not),regardless of the definition of the risk population.Specifically,the risk of co-occurrence of de-pressive symptoms and overweight or obesity was reduced by 33％(95％CI:14％to 48％,based on the average level across multiple measurements over time)to 78％(95％CI:6％to 95％,based on the joint trajectories of depressive symptoms and BMI over time)per 1-standard deviation(1-SD)increase in self-control score.In addition,the risk of depressive-symptom-dominant and overweight-or-obesity-dominant was reduced by 25％(95％CI:4％to 42％,only based on the average level across multiple measure-ments over time)and 21％(95％CI:1％to 37％,only based on the joint trajectories of depressive symptoms and BMI over time)per 1-SD increase in self-control score,respectively.The results from sen-sitivity analysis that defined individuals'weight status according to World Health Organization(WHO)standards were consistent with our main findings.Conclusion:Individuals with higher self-control scores from adolescence to early adulthood have a lower risk of co-occurrence of depressive symptoms and over-weight or obesity,suggesting that personalized interventions for co-occurrence of depressive symptoms and overweight or obesity can be carried out based on self-control scores in the future.

Objective:To analyze the correlation between clinical features and prognosis or prognostic risk factors in patients with KMT2A::AFF1 gene positive B-ALL.Methods:Retrospective cohort study was conducted. 167 cases of B-ALL admitted to the Shanghai General Hospital and the Naval Medical University Affiliated First Hospital from April 1, 2011 to July 31, 2022 were divided into groups according to gene types. 22 cases with KMT2A::AFF1 positive B-ALL were enrolled as the experimental group, 54 cases with BCR::ABL gene positive B-ALL as control group 1 and 91 cases with KMT2A::AFF1 and BCR::ABL gene negative B-All as control group 2. The median age of first diagnosis in the experimental group, control group 1 and control group 2 were 43.5(30.5, 56), 43.5(34, 55) and 32(24, 46) respectively. The median white blood cell counts of the three groups were 142.4(25.7, 247.2)×10 9/L, 37.6(15.7, 102.2)×10 9/L and 13.4(4.3, 33.0)×10 9/L, respectively. Allo-HSCT rates in three groups were 45.5%, 72.2% and 72.5% respectively. Using SPSS 26.0 software, the statistical methods of nonparametric rank sum test, chi-square test, Kaplan-Meier and Cox regression were used to analyze and compare the differences in clinical characteristics, chemotherapy and prognosis between the experimental group and the control groups, and to analyze the risk factors and the differences in prognosis of allo-HSCT in the experimental group. Results:The age difference between the experimental group and the control group 2 was significant ( Z=-2.151, P=0.031). The white blood cell count in experimental group was significantly higher than that in control group 1 ( Z=-2.363, P=0.018) and control group 2 ( Z=-4.886, P<0.001). The rate of allo-HSCT in experimental group was lower than that in control group 1(45.5% vs 72.2%, χ 2=4.890, P=0.027) and control group 2 (45.5% vs 72.5%, χ 2=5.897, P=0.015). The remission rates of the patients in three groups after receiving one course of chemotherapy were 60%(12/20), 83.3%(45/54) and 76.6%(69/90); the remission rates after two courses of chemotherapy were 25%(5/20), 7.4%(4/54) and 12.2% (11/90), and the non-remission rates of more than two courses of treatment were 15%(3/20), 9.3%(5/54) and 11.1%(10/90), respectively. The effect of chemotherapy in experimental group was worse than that in control group 1 ( Z=-1.979, P=0.048). There was no significant difference between the three groups in sex, whether the chromosome is a standard-risk karyotype, hemoglobin at the time of initial onset, platelet count and percentage of bone marrow blast cells. The overall survival rate (OS) of experimental group was significantly lower than that of control group 1 and control group 2(23.9% vs 36.7%, χ 2=7.608, P=0.006 and 23.9% vs. 44.8%, χ 2=6.442, P=0.011), and the 3-year recurrence-free survival (RFS) was also lower than that of the other two groups (14.0% vs 57.6%, χ 2=17.823, P<0.001 and 14.0% vs 48.2%, χ 2=16.432, P<0.001). There was a significant difference in the total OS rate between the experimental group and the group without allo-HSCT (45.0% vs 9.2%, χ 2=15.254, P<0.001). Univariate analysis showed that age was the risk factor of RFS in the experimental group, and allo-HSCT therapy was the protective factor of OS. Multivariate analysis showed that allo-HSCT was an independent protective factor for OS in the experimental group. Conclusions:Patients with KMT2A::AFF1 positive B-ALL had higher white blood cells, less sensitivity to chemotherapy and poor prognosis. Age was a risk factor of RFS in KMT2A::AFF1 positive B-ALL, and allo-HSCT could improve the prognosis.

Objective To retrieve,appraise and integrate the relevant evidence of non-pharmacological sleep management measures in critically ill children.Methods We conducted a systematical search on the topic of non-pharmacological sleep management measures in critically ill children across multiple databases,including BMJ best practice,UpToDate,International Guidelines International Network,National Institute for Health and Care Excellence,Scottish Intercollegiate Guidelines Network,Registered Nurses'Association of Ontario,Joanna Briggs Institute Library,the American Academy of Sleep Medicine,International Pediatric Sleep Association,CNKI,WanFang database,VIP database,SinoMed,Cochrane Library,PubMed,Web of Science,and CINAHL.The literature retrieval time limit was from the establishment of the database to February 2023.The study design includes clinical practice guideline,evidence summary,clinical decision-making,expert consensus,and systematic review.The evidence was extracted and summarized according to the subject after the independent literature quality evaluation by 2 researchers.Results 10 relevant pieces of literature were identified,including 2 guidelines,1 consensus statement,4 systematic reviews,2 evidence summaries,and 1 top clinical decision.These sources provided a total of 28 pieces of evidence across 5 key themes,including sleep assessment,management principles and education,environmental management and sleep promotion.Conclusion This study summarizes the best available evidence on non-pharmacological sleep management measures in critically ill children.It is recommended that healthcare professionals should consider the clinical context when implementing evidence-based interventions,aiming to reduce nocturnal awakenings,extend sleep duration,and improve sleep quality in critically ill children.

Objective:To observe and analyze the morphological characteristic of bone marrow and peripheral blood in patients diagnosed with de novo acute leukemia.Methods:From October 1, 2015 to December 31, 2021, 1151 patients aged 47 (26, 62) years, consisting of 602 males and 549 females with newly diagnosed acute leukemia in the Department of Hematology, Affiliated Hospital of Xuzhou Medical University, were collected to preform the morphological analysis in bone marrow and peripheral blood smears. Based on the comprehensive diagnosis results of morphology, immunology, cytogenetics, and molecular biology, comparison between acute myeloid leukemia (AML) and acute lymphoblastic leukemia (ALL), AML with RUNX1-RUNXITI gene, AML with CBFβ/MYH11 gene, acute promyelocytic leukemia (APL) with PML/RARA gene, AML with NPM1 gene, the rest of the AML, Ph+ALL and Ph-ALL were performed by Chi-square test along with analysis of the differences in the ratio of wood bundle cells, pseudo-Chediak-Higashi (PCH) inclusions, cytoplasmic small particles, nuclear notches, leukemia cells with cup-like changes (cup cells); as well as the differences in the micromeganuclei, early immature granulocytes, plasma cells, high eosinophils and other accompanying cells and the distribution of "grape-like" aggregation. Finally, the morphological characteristics of acute leukemia cells, the appearance and arrangement of accompanying cells were summarized.Results:Between AML and ALL, there were statistically significant differences in cytoplasmic Auer bodies[(45.5%, 0%), χ 2=211.400, P<0.01], PCH inclusion bodies[(28.9%, 0%), χ 2=114.100, P<0.01], cytoplasmic fine particles[(20.7%, 2.9%), χ 2=53.798, P<0.01], nuclear notches[(0.7%, 6.1%), χ 2=30.906, P<0.01], and goblet cells[(4.9%, 0.3%), χ 2=13.495, P<0.01], micromegakaryus [(22.4%, 0.3%), χ 2=80.398, P<0.01], plasma cells[(87.6%, 10.6%), χ 2=604.241, P<0.01], hyperacidophils[(15.3%, 1.0%), χ 2=46.116, P<0.01] showed significant differences in the "grape-like" aggregation distribution. In AML with RUNX1-RUNXITI gene, the changes of vacuoles and PCH inclusion bodies are more obvious; in AML with CBFβ/MYH11 gene, the increase of hypereosinophils is more obvious; in APL with PML/RARA gene, the increase of woodbundle is more obvious. The morphology of nuclei chromatin, nucleolus, and vacuoles were also different among the groups. Comparison between Ph+ALL and Ph-ALL showed that Ph+ALL was more prone to develop early immature granulocytes and plasma cells (all P<0.05). Conclusion:There are significant differences between AML and ALL in the characteristics of leukemia cells, the regularity of accompanying cells, and the aggregation and distribution patterns. The subtypes of AML with specific genetic abnormalities have their own characteristics in the appearance of vacuoles, PCH inclusions, hypereosinophils, woodbundle cells, and goblet cells. Ph+ALL is more prone to present early immature granulocytes and plasma cells.

ObjectiveTo study the changes of primary metabolites and phenols in the fruits of Acanthopanax senticosus at different development stages, so as to provide a theoretical basis for the rational utilization of A. senticosus fruit resources. MethodThe primary metabolites and phenols in the fruits at different development stages were determined via gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-mass spectrometry (LC-MS) and then compared by multivariate statistical analysis. ResultA total of 274 chromatographic peaks were obtained by GC-MS-based non-targeted metabonomics and 24 differential metabolites were screened out by multivariate statistical analysis. The differential metabolites were mainly concentrated in pentose phosphate pathway, galactose metabolism, ascorbic acid and aldose metabolism pathways. After color conversion, the pentose phosphate pathway and galactose metabolism were activated and increasing sugars were accumulated. The ascorbic acid and aldose metabolism pathways were active before color conversion, with high accumulation of the end product ascorbic acid. The ultra-high liquid chromatography-mass spectrometry (UPLC-MS) identified 28 phenols in the fruits at different development stages. Flavonoids were accumulated mainly at the green ripening stage before color conversion, and phenolic acids were accumulated mainly after color conversion. ConclusionThe accumulation of primary metabolites and phenols in A. senticosus fruits varies significantly among different development stages

Our endocrinology teaching group has carried out the application and exploration of mobile APP teaching mode based on Superstar learning. Through the questionnaire survey, students thought there was no significant difference between mobile APP teaching and traditional teaching in knowledge understanding and memory. But the classroom atmosphere of mobile APP teaching was relaxed and active, which helps to improve learning interest and self-learning ability, and the problems after class can be solved by feedback in time. Through the sharing of experience on the whole course management in endocrinology mobile APP teaching, it is expected to provide positive reference for the mobile teaching mode of medical education in the future.

Objective:To explore the evaluation dimensions and indicators of research hospital, provide empirical evidence for the construction of research hospital.Methods:Non-probability sampling was adopted, physicians from 9 hospitals in Shanghai that have a National Clinical Medical Research Center or Shanghai Clinical Medical Research Center were invited as survey participants. The physician participants were divided into senior and junior groups. For senior group, a questionnaire including 5 dimensions and 21 indicators was provided. The survey participants were consulted to determine whether the indicators can be used to measure this dimension, and also invited to propose additional dimensions and indicators for improvement.For junior group, two open questions were proposed to explore their needs of support in both clinical service and research.Results:Based on suggestions from the research participants, the research team, and other expert consultant, this article tried to propose 5 dimensions and 21 indicators for evaluating research hospitals. Among them, 11 were indicators compiled by the research team based on the literature review and agreed by more than 2/3 of senior physicians surveyed, while others were proposed based on the survey results. The research team planned to use the entropy method to determine the weights of different indicators, thus, the participants were not required to make judgments on the weights of dimensions and indicators.Conclusions:The evaluation of research hospitals has not yet formed a systematic consensus. Through the exploration and establishment of evaluation dimensions and indicators, identify hospitals that are closer to the development goals of research hospitals can provide a basis for future next-step practices.

Objective:To study the basic characteristics and gene evolution of M segment of fever and thrombocytopenia syndrome virus (SFTSV)in China.Methods:The full sequence of M fragment of SFTSV strain isolated in China before June 25, 2020 was obtained from GenBank. The BioEdit software was used for multi-sequence alignment and MEGA5.0 software was used to construct the phylogenetic tree to compare the homology of SFTSV, which came from different types and sources.Results:M fragments of 203 SFTSV isolated in China was collected from GenBank database.. Among them, 185 strains were isolated from humans (91.1%), 11 strains were isolated from ticks(5.4%), and 5 strains were isolated from goat, mice and hedgehog (3.5%). Phylogenetic tree analysis showed that C2 genotype was dominant in China, accounting for 42.4% of total. The nucleotide sequence and amino acid sequence homology were 96.4%~100.0%, 94.5%~100.0%, respectively. The homology of human and animal isolates was very high. The nucleotide sequence and amino acid sequence homology were 92.5%~100.0% and 92.0%~100.0%, respectively.Conclusions:Phylogenetic tree analysis of SFTSV isolates in China based on the M fragment showed that C2 genotype was dominant, and there are multiple types of co-circulation. By comparing M and S-segment-based phylogenetic tree, it suggests that SFTSV may potential undergo adaptive conversion from C-type to J-type.