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 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.

Objective To study the diagnostic and therapeutic value of endoscopic irrigation for uncomplicated acute colonic diverticulitis.Methods From June 2021 to March 2022,4 patients suspected uncomplicated acute colonic diverticulitis who then underwent colonoscopy,and endoscopic irrigation treatment was performed after confirming.The endoscopic imaging of acute colonic diverticulitis was summarized.The changes of abdominal pain symptoms and prognosis of the patients were evaluated.The levels of white blood cell(WBC)and C-reactive protein(CRP)in serum before and after treatment were measured.Results 4 patients were diagnosed as uncomplicated acute colonic diverticulitis underwent colonoscopy.Two cases were located in the ascending colon and the other two in the cecum.Endoscopic imaging characteristics included mucosal hyperemia and swelling of diverticular orifice and nearby area,purulent secretions,purulent fur and fecalith at the diverticular orifice.After treatment,the symptom of abdominal pain was relieved,the levels of WBC and CRP in serum were lower than before treatment.Conclusion Endoscopic irrigation has a useful diagnostic and therapeutic value for uncomplicated acute diverticulitis of the colon.It is worthy of further clinical study.

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).

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 evaluate the quality of the clinical trial literatures published on the treatment of COVID-19 with TCM, find out the shortcomings and put forward corresponding suggestions, in order to promote TCM against COVID-19. Methods "COVID-19", "New Coronavirus Pneumonia", "TCM", "Chinese Medicine", "Clinical Trial", "Lianhua Qingwen", "Huoxiang Zhengqi", "Jinhua Qinggan", and other keywords were used to search relevant literatures in CNKI and PubMed database. Among the all the screened relevant literatures on the treatment of COVID-19 with TCM, the literature quality was assessed according to evaluation criteria of clinical trial literatures. Results A total of 463 papers related to the treatment of COVID-19 with TCM were obtained. 440 papers on theoretical research on the network pharmacology mechanism of Chinese medicine treatment of COVID-19 were excluded. Among the 23 articles included in the quality analysis, there are 3 randomized controlled studies, 1 multi-center prospective randomized controlled trial, 2 disease case report trials, and 5 uncontrolled single arm studies, 5 controlled trials and 7 retrospective studies. In the period of more than half a year, although many clinical trial documents of TCM for the treatment of COVID-19 have been published, the clinical trial design reflected in most TCM clinical trials were not standardized. There are problems in "randomness and rationality", such as no control group, no randomization design, only case studies, no blinding method in controlling bias, and insufficient objectivity in the evaluation criteria of efficacy. All of these need to be improved. Conclusion The treatment of new coronavirus pneumonia with TCM still requires more and standardized clinical trial verifications and publications to generate strong evidence-based results, such as adding control groups, increasing sample size, and using blinding methods to increase the credibility of clinical trials.

Objective:To screen the differentially expressed genes on whole expression profiles of the inflammation-related cytokines in mice infected with influenza virus by the gene chip technology, and to explore the intervention effect of Shufeng Xuanfei Jiedu formula.Methods:Male ICR mice were divided into normal group (N group), influenza virus infective model group (M group), Oseltamivir control group (C group) and Shufeng Xuanfei Jiedu formula high, medium and low dose groups (SH, SM, SL groups) according to the random number table method, with 10 rats in each group. A mouse model of influenza virus pneumonia was reproduced by nasal drip of influenza virus strain FM1 (0.05 mL). In N group, 0.05 mL normal saline was used. In SH, SM and SL groups, Shufeng Xuanfei Jiedu formula was used 2 hours after intranasal infection (2 times, equal and 1/2 of the clinical treatment dose, approximately 3.8, 1.9 and 1.0 g·mL -1·d -1) for 4 days. In C group, the dosage of Oseltamivir was 2.5 g·mL -1·d -1. In N group and M group, distilled water was given (0.2 mL once a day). On the 5th day, the whole lung of mice was taken. The lung index was calculated, and the pathological sections were observed. The total RNA of lung tissue was extracted and detected after hybridization with mice whole gene expression spectrum chip to select differentially expressed genes of chemokine pathways. The expression intensity ratio of the chip probe signal in each group vs. M group was calculated, and P < 0.05 and log 2ratio > 1 were up-regulated genes, while P < 0.05 and log 2ratio < -1 were down-regulated genes. Results:Compared with the N group, the lung index in the M group was significantly higher, and pathological changes were found in lung tissue, which suggested that the model of influenza virus infection was successfully established. Compared with the M group, the lung index of mice in C, SH, SM, SL groups was significantly lower (0.96±0.14, 1.45±0.22, 1.14±0.18, 1.22±0.21 vs. 1.72±0.15, all P < 0.05), and the extent and degree of lesions were reduced, however, there was no significant difference among the groups. Gene chip analysis showed that there were more differentially expressed genes in N group vs. M group, SH group vs. M group, SM group vs. M group, SL group vs. M group. It could be used for further signal transduction pathway screening. Compared with N group, the differential gene expression of chemokine C-C ligands (CCL-3, CCL-5) and chemokine C-X-C ligands (CXCL-9, CXCL-10) in M group were significantly up-regulated [log 2 (M group/N group) were 6.64, 3.51, 5.40, 6.64, respectively]. Compared with M group, the gene expressions of CCL-3, CCL-5, CXCL-9 and CXCL-10 were significantly down-regulated in C, SH, SM and SL groups [log 2 (C group/M group) were -3.96, -2.26, -3.12, -2.40; log 2 (SH group/M group) were -5.57, -2.37, -1.57, -1.01; log 2 (SM group/M group) were -4.35, -1.47, -1.26, -1.74; log 2 (SL group/M group) were -2.86, -1.86, -1.23, -1.39, respectively]. Conclusion:Shufeng Xuanfei Jiedu formula inhibits inflammatory damage in mice after influenza virus infection by down-regulating the expressions of CCL-3, CCL-5, CXCL-9 and CXCL-10 on chemokine pathways.

The new type of coronavirus pneumonia referred to as coronavirus disease 2019 (COVID-19), became pandemic globally, causing tens of millions of infections. The establishment of a sound prevention and control system is particularly important for disease prevention and control. In the process of admission and treatment of COVID-19 patients, we have established an effective rapid prevention and control system for special infections in the hospital. Through the establishment of a hospital-level emergency response and first aid training system, the closed-loop communication between hospitals, departments and medical care is quick to respond. We categorize treatment according to patients’ condition, provide rapid feedback and return, implement the specific responsibility system for special personnel, improve the rapid prevention and control system for special infections, and apply it to the prevention and control of the epidemic of COVID-19 patients in the region, and achieve the effectiveness of the anti-epidemic through comprehensive management measures. The control has achieved zero infection among medical staff and within the hospital, ensuring that the hospital can quickly resume daily work at the end of the epidemic, and has certain promotion and demonstration value.

【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.