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 analyze the changes in the disease spectrum of hospitalized patients with neurological diseases in a tertiary hospital in the past 5 years,in order to effectively develop targeted disease prevention and treatment strategies.Methods Collect case data of neurological diseases in a tertiary hospital from January 1,2018 to December 31,2022,and use ICD-10 coding for disease classification,and analyze the disease type,gender,age groups,and other factors.Results A total of 9060 patients with neurological diseases were admitted in the past five years.In 2020,the number of discharged patients affected by the COVID-19 was the smallest,accounting for 15.96% ,and in 2022,the number was the largest,accounting for 24.05% .The number of cases showed an increasing trend.There was no statistically significant difference in the composition of male and female patients;There is a statistically significant difference in the number of cases among patients of different age groups,and the incidence categories are different;The top 10 diseases in the ranking of neurological diseases are:transient ischemic attack,headache,spinal nerve disease,neurological disorders,epilepsy,Parkinson's disease,sleep disorders,paralysis,other brain diseases,hydrocephalus.The distribution of the top 10 diseases by gender has statistical significance.Conclusion The hospital can formulate the diagnosis and treatment technology for different age groups according to the distribution characteristics of disease spectrum,carry out comprehensive prevention and treatment measures for key groups,strengthen the construction of key special-ties and allocate health resources properly.

In order to develop a positive quality control products for the detection of porcine repro-ductive and respiratory syndrome virus(PRRSV)nucleic acid by real-time fluorescent quantitative PCR(RT-qPCR),the positive quality control products of PRRSV-1 and PRRSV-2 M genes were prepared using armored RNA technology of MS2 phage.PRRSV-1 and PRRSV-2 M genes were amplified,purified and recovered,and ligated into pET28b vector containing MS2 mature enzyme protein gene and capsid protein.After transformed into BL21(DE3),the gene products were in-duced by IPTG and purified by PEG6000 precipitation method to prepare the armored RNA virus-like particles(AR-PRRSV)containing PRRSV M gene.Following the performance evaluation,as the positive quality control products of PRRSV-1 and PRRSV-2 M genes,AR-PRRSV1M and AR-PRRSV2M were calculated using YY/T 1652-2019 standard.Results showed that it had a good u-niformity,stable storage for the armored virus-like particles at-20,4,25 ℃ for 60 d,and 37 ℃ for 30 d.The prepared armored virus-like particles AR-PRRSV1M and AR-PRRSV2M were deter-mined by digital quantitative PCR(ddPCR)after preliminary quantification by RT-qPCR.The 104 copies/μL of AR-PRRSV1M and AR-PRRSV2M ddPCR fixation was(1.33+0.50)× 104 cop-ies/μL.The above results indicates that the AR-PRRSVM can be used as the quality control of the whole detection process(nucleic acid extraction,reverse transcription and RT-qPCR).

Objective To investigate the dosimetric difference between manual and inverse optimization in 3-dimensional (3D) brachytherapy for gynecologic tumors. Methods This retrospective study was conducted among a total of 110 patients with gynecologic tumors undergoing intracavitary combined with interstitial brachytherapy or interstitial brachytherapy. Based on the original images, the brachytherapy plans were optimized for each patient using Gro, IPSA1, IPSA2 (with increased volumetric dose limits on the basis of IPSA1) and HIPO algorithms. The dose-volume histogram (DVH) parameters of the clinical target volume (CTV) including V200, V150, V100, D90, D98 and CI, and the dosimetric parameters D2cc, D1cc, and D0.1cc for the bladder, rectum, and sigmoid colon were compared among the 4 plans. Results Among the 4 plans, Gro optimization took the longest time, followed by HIPO, IPSA2 and IPSA1 optimization. The mean D90, D98, and V100 of HIPO plans were significantly higher than those of Gro and IPSA plans, and D90 and V100 of IPSA1, IPSA2 and HIPO plans were higher than those of Gro plans (P<0.05), but the CI of the 4 plans were similar (P>0.05). For the organs at risk (OARs), the HIPO plan had the lowest D2cc of the bladder and rectum;the bladder absorbed dose of Gro plans were significantly greater than those of IPSA1 and HIPO (P<0.05). The D2cc and D1cc of the rectum in IPSA1, IPSA2 and HIPO plans were better than Gro (P<0.05). The D2cc and D1cc of the sigmoid colon did not differ significantly among the 4 plans. Conclusion Among the 4 algorithms, the HIPO algorithm can better improve dose coverage of the target and lower the radiation dose of the OARs, and is thus recommended for the initial plan optimization. Clinically, the combination of manual optimization can achieve more individualized dose distribution of the plan.

Objective To investigate the dosimetric difference between manual and inverse optimization in 3-dimensional (3D) brachytherapy for gynecologic tumors. Methods This retrospective study was conducted among a total of 110 patients with gynecologic tumors undergoing intracavitary combined with interstitial brachytherapy or interstitial brachytherapy. Based on the original images, the brachytherapy plans were optimized for each patient using Gro, IPSA1, IPSA2 (with increased volumetric dose limits on the basis of IPSA1) and HIPO algorithms. The dose-volume histogram (DVH) parameters of the clinical target volume (CTV) including V200, V150, V100, D90, D98 and CI, and the dosimetric parameters D2cc, D1cc, and D0.1cc for the bladder, rectum, and sigmoid colon were compared among the 4 plans. Results Among the 4 plans, Gro optimization took the longest time, followed by HIPO, IPSA2 and IPSA1 optimization. The mean D90, D98, and V100 of HIPO plans were significantly higher than those of Gro and IPSA plans, and D90 and V100 of IPSA1, IPSA2 and HIPO plans were higher than those of Gro plans (P<0.05), but the CI of the 4 plans were similar (P>0.05). For the organs at risk (OARs), the HIPO plan had the lowest D2cc of the bladder and rectum;the bladder absorbed dose of Gro plans were significantly greater than those of IPSA1 and HIPO (P<0.05). The D2cc and D1cc of the rectum in IPSA1, IPSA2 and HIPO plans were better than Gro (P<0.05). The D2cc and D1cc of the sigmoid colon did not differ significantly among the 4 plans. Conclusion Among the 4 algorithms, the HIPO algorithm can better improve dose coverage of the target and lower the radiation dose of the OARs, and is thus recommended for the initial plan optimization. Clinically, the combination of manual optimization can achieve more individualized dose distribution of the plan.

The formation of learning and memory is regulated by synaptic plasticity in hippocampal neurons. Here we explored how gestational exposure to dexamethasone, a synthetic glucocorticoid commonly used in clinical practice, has lasting effects on offspring's learning and memory. Adult offspring rats of prenatal dexamethasone exposure (PDE) displayed significant impairments in novelty recognition and spatial learning memory, with some phenotypes maintained transgenerationally. PDE impaired synaptic transmission of hippocampal excitatory neurons in offspring of F1 to F3 generations, and abnormalities of neurotransmitters and receptors would impair synaptic plasticity and lead to impaired learning and memory, but these changes failed to carry over to offspring of F5 and F7 generations. Mechanistically, altered hippocampal miR-133a-3p-SIRT1-CDK5-NR2B signaling axis in PDE multigeneration caused inhibition of excitatory synaptic transmission, which might be related to oocyte-specific high expression and transmission of miR-133a-3p. Together, PDE affects hippocampal excitatory synaptic transmission, with lasting consequences across generations, and CDK5 in offspring's peripheral blood might be used as an early-warning marker for fetal-originated learning and memory impairment.

Objective:To explore the construction of evaluation index system of core competence of imaging specialized nurses and analysis of reliability and validity test.Methods:Firstly, a research group was established to consult the relevant literature. After repeated discussion among the members of the research group, imaging specialized nurses were invited to discuss, modify and improve the framework of the evaluation index system, and an expert consultation questionnaire for the evaluation index system of the core competence of imaging specialized nurses was formulated. Through two rounds of expert consultation, the core competence evaluation index system of imaging specialized nurses was constructed, and a questionnaire was compiled based on the constructed index system. Questionnaire survey method was used to verify the rationality and scientificity of the core competence evaluation index system of imaging specialized nurses constructed in this study.Results:The evaluation index system of core competence of imaging specialized nurses included 3 first-level indexes, 20 second-level indexes and 48 third-level indexes. The overall Cronbach's α coefficient of the evaluation index system was 0.972. The Cronbach's α coefficients of the theoretical knowledge of imaging specialized nursing, nursing practice skills of imaging specialized nurses and related abilities were 0.915, 0.934 and 0.924 respectively. The overall content validity index (S-CVI) of the index system was 0.977, and the CVI of each item (I-CVI) was 0.812-1.000. The overall correlation coefficient of the first-level indexes and index system was 0.800-0.936 ( P < 0.05), the correlation coefficient of the secondary indexes and the corresponding primary indexes was 0.562-0.972 ( P<0.05), and the correlation coefficient of the third-level indexes and the corresponding first-level index was 0.362-0.914 ( P<0.05). Conclusion:The constructed questionnaire for evaluation index of core competence of imaging specializied nurses has good reliability and consistent internal structure, and has good feasibility, reliability, rationality and scientificity, which provides a theoretical basis and practical guide for the cultivation, assessment and evaluation of core competence of imaging specialized nurses.

In the emergency of the outbreak of COVID-19 in December 2019, Shaanxi Provincial Health Committee mobilized several medical teams from major hospitals in the province, and, by relying on Xi’an Chest Hospital, jointly established an anti COVID-19 consortium to control and eradicate the epidemic in a short time. Information support is an important guarantee for winning this battle. In order to realize the efficient cooperation among multiple medical teams, we have carried out some exploratory and innovative information support services on the basis of the original information system of the chest hospital. In this process, we have gone through some detours. Some compromises were made on some problems that could not be solved in the short term. Finally, in an environment full of uncertainty, a set of information support management system with basically smooth operation was built through rapid trial and error adjustment. The system mainly includes the following aspects: support of the organizational structure and operation process of the anti-epidemic consortium, support for medical collaboration related businesses of multiple medical teams, and support for statistical reports and online meetings. Information support has played a very important role in this action, and this practice has also accumulated experience for us to deal with similar situations in the future.

Objective:To investigate the prognostic value of tumor infiltrating lymphocytes (TILs) and their phenotypes, CD4 + TILs, CD8 + TILs and FOXP3 + TILs, in short-term prognosis (PCR) and long-term prognosis (DFS, OS) of triple negative breast cancer (TNBC), and to provide theoretical support for the immunotherapy of TNBC. Methods:A systematic surch of PubMed, Medline, Embase, Web of Science, CBM, CNKI was conducted to identified eligible articles. All prospective trials and observational controlled trials published before August 2020 were retrieved.The two authors independently evaluated the quality of the included literature, extracted the data, and conducted systematic evaluation and analysis using RevMan5.3 software.Results:A total of 48 literatures were included.Every 10% increase in TILs and TILs expression in the high-expression group predicted a higher PCR, OR were 1.81 (95% CI: 1.44-2.28) and 1.09 (95% CI: 1.02-1.15). For longer survival, the HR for DFS and OS were 0.99 (95% CI: 0.98-0.99) and 0.90 (95% CI: 0.86-0.94), 0.87 (95% CI: 0.79-0.96) and 0.88 (95% CI: 0.83-0.92). Phenotypic CD4 + TILs, CD8 + TILs, and FOXP3 + TILs did not show statistically significant differences in PCR among patients, but in long-term prognosis, higher infiltration predicted longer survival, with HR for DFS of 0.54 (95% CI: 0.36-0.80), 0.46(95% CI: 0.33-0.64) and 0.45(95% CI: 0.31-0.68). HR of OS were 0.49 (95% CI: 0.32-0.76), 0.59 (95% CI: 0.54-0.66) and 0.44 (95% CI: 0.27-0.71). Conclusion:The infiltration degree of TILs and the increase of TILs expression by 10% can be used as a prognostic index of TNBC.High levels of phenotypic infiltration of CD4 + TILs, CD8 + TILs, and FOXP3 + TILs predicted better long-term survival, but there was no statistically significant difference in short-term survival.

【Objective】 To understand the current status of medical staff’s awareness of hospital infection prevention and control during the epidemic of COVID-19 in Shaanxi Province and analyze its influencing factors. 【Methods】 The questionnaire was designed on the "Questionnaire Star" website. Based on the WeChat platform, a voluntary sampling method was used to invite online questionnaires. From March 13 to 29, we collected a total of 8037 questionnaires, 30 of which did not meet the requirements and had logical problems, and finally 8 007 valid questionnaires were obtained. 【Results】 A total of 8 007 medical staff were surveyed. Among them, Medical staff of Grade 3A, Grade 3B, Grade 2A, and Grade 2B hospital accounted for 39.6%, 2.3%, 55.3%, and 2.6%, respectively. The average age of the respondents was (32.1±7.2) years old, including 7 199 nurses and 501 doctors. The training effect was statistically significant in different regions, different hospital levels, whether it was a designated hospital and whether there were sensor control supervisors, as well as medical staff of different specialty, professional title, and work area (all P<0.01). The basic cognitive situation of the surveyed subjects showed that most medical staff in Shaanxi Province had a correct rate of less than 30% in whether they needed to disinfect before de-protection and how to disinfect the hospital environment. Multivariate linear regression analysis showed that the cognition level of medical staff in Grade A hospitals was significantly lower than that in Grade A hospitals (P<0.01). The cognition level of medical staff in designated hospitals was significantly higher than that of others (P<0.01). The cognition level of medical staff in hospitals with sensory control supervisors was significantly higher than that of others (P<0.01). The cognition level of people aged 25-34 and 35-44 was significantly lower than those aged 45 and above (all P<0.01). The cognition level of medical technicians and service personnel was significantly lower than that of doctors (P=0.02 and <0.01, respectively). The cognition level of medical staff with intermediate, associate senior, and senior professional titles was significantly higher than the cognition level of those with junior and below professional titles (all P<0.01). The cognition level of medical staff in fever clinics, emergency departments, isolation wards, ICU and other surgeries was significantly higher than that of those working in ordinary outpatient department (P=0.01, 0.03, <0.01, 0.02, and <0.01 respectively). 【Conclusion】 Most medical staff in Shaanxi Province have misunderstandings about whether they need to disinfect before de-protection and how to disinfect the hospital environment. Moreover, we found that the awareness of medical staff in Shaanxi Province of hospital infection prevention and control during the epidemic of COVID-19 was affected by the hospital’s level, whether it was a designated hospital, whether there were sensor control supervisors, as well as the age, specialty, professional title and work area of the medical staff.