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*

Background/Aims: The World Health Organization set the goal of eliminating hepatitis C virus (HCV) by 2030, with 80% and 65% reductions in HCV incidence and mortality rates, respectively. We aimed to evaluate the health benefits, cost-effectiveness and return on investment (ROI) of HCV elimination. Methods: Using an HCV transmission compartmental model, we evaluated the benefits and costs of different strategies combining screening and treatment for Chinese populations. We identified strategies to achieve HCV elimination and calculated the incremental cost-effectiveness ratios (ICERs) per disability-adjusted life year (DALY) averted for 2022–2030 to identify the optimal elimination strategy. Furthermore, we estimated the ROI by 2050 by comparing the required investment with the economic productivity gains from reduced HCV incidence and deaths. Results: The strategy that results in the most significant health benefits involves conducting annual primary screening at a rate of 14%, re-screening people who inject drugs annually and the general population every five years, and treating 95% of those diagnosed (P14-R4-T95), preventing approximately 5.75 and 0.44 million HCV infections and deaths, respectively, during 2022–2030. At a willingness-to-pay threshold of $12,615, the P14-R4-T95 strategy is the most cost-effective, with an ICER of $5,449/DALY. By 2050, this strategy would have a net benefit of $120,997 million (ROI=0.868). Conclusions Achieving HCV elimination in China by 2030 will require significant investment in large-scale universal screening and treatment, but it will yield substantial health and economic benefits and is cost-effective.

Background/Aims: The World Health Organization set the goal of eliminating hepatitis C virus (HCV) by 2030, with 80% and 65% reductions in HCV incidence and mortality rates, respectively. We aimed to evaluate the health benefits, cost-effectiveness and return on investment (ROI) of HCV elimination. Methods: Using an HCV transmission compartmental model, we evaluated the benefits and costs of different strategies combining screening and treatment for Chinese populations. We identified strategies to achieve HCV elimination and calculated the incremental cost-effectiveness ratios (ICERs) per disability-adjusted life year (DALY) averted for 2022–2030 to identify the optimal elimination strategy. Furthermore, we estimated the ROI by 2050 by comparing the required investment with the economic productivity gains from reduced HCV incidence and deaths. Results: The strategy that results in the most significant health benefits involves conducting annual primary screening at a rate of 14%, re-screening people who inject drugs annually and the general population every five years, and treating 95% of those diagnosed (P14-R4-T95), preventing approximately 5.75 and 0.44 million HCV infections and deaths, respectively, during 2022–2030. At a willingness-to-pay threshold of $12,615, the P14-R4-T95 strategy is the most cost-effective, with an ICER of $5,449/DALY. By 2050, this strategy would have a net benefit of $120,997 million (ROI=0.868). Conclusions Achieving HCV elimination in China by 2030 will require significant investment in large-scale universal screening and treatment, but it will yield substantial health and economic benefits and is cost-effective.

Background/Aims: The World Health Organization set the goal of eliminating hepatitis C virus (HCV) by 2030, with 80% and 65% reductions in HCV incidence and mortality rates, respectively. We aimed to evaluate the health benefits, cost-effectiveness and return on investment (ROI) of HCV elimination. Methods: Using an HCV transmission compartmental model, we evaluated the benefits and costs of different strategies combining screening and treatment for Chinese populations. We identified strategies to achieve HCV elimination and calculated the incremental cost-effectiveness ratios (ICERs) per disability-adjusted life year (DALY) averted for 2022–2030 to identify the optimal elimination strategy. Furthermore, we estimated the ROI by 2050 by comparing the required investment with the economic productivity gains from reduced HCV incidence and deaths. Results: The strategy that results in the most significant health benefits involves conducting annual primary screening at a rate of 14%, re-screening people who inject drugs annually and the general population every five years, and treating 95% of those diagnosed (P14-R4-T95), preventing approximately 5.75 and 0.44 million HCV infections and deaths, respectively, during 2022–2030. At a willingness-to-pay threshold of $12,615, the P14-R4-T95 strategy is the most cost-effective, with an ICER of $5,449/DALY. By 2050, this strategy would have a net benefit of $120,997 million (ROI=0.868). Conclusions Achieving HCV elimination in China by 2030 will require significant investment in large-scale universal screening and treatment, but it will yield substantial health and economic benefits and is cost-effective.

BACKGROUND:There is still no consensus on the optimal internal fixation for the treatment of Pauwels Ⅲ femoral neck fracture,and most of the related finite element analyses have been performed using a single simplified loading condition,and the biomechanical properties of commonly used internal fixation devices need to be further investigated. OBJECTIVE:To analyze the biomechanical characteristics of Pauwels Ⅲ femoral neck fractures treated with cannulated compression screw,dynamic hip screw,and femoral neck system by finite element method under different loading conditions of single-leg standing loads and sideways fall loads. METHODS:The DICOM data of healthy adult femur were obtained by CT scanning,imported into Mimics 15.0 software to obtain the rough model of bone tissue.The data exported from Mimics were optimized by Geomagics software,and then three internal fixation models were built and assembled with the femur model according to the parameters of the clinical application of the cannulated compression screw,dynamic hip screw,and femoral neck system by using Pro/E software.Finally,the three internal fixation models were imported into Ansys software for loading and calculation to analyze the stress distribution and displacement of the femur and the internal fixation under different working conditions of single-leg standing loads and sideways fall loads,as well as the stress characteristics of the calcar femorale and Ward's triangle. RESULTS AND CONCLUSION:(1)Under the single-leg standing load and the sideways fall load,the proximal femoral stress of the three internal fixation models was mainly distributed above the fracture end of the femoral neck.The peak stress of the proximal femoral end,fracture end,Ward triangle,and calcar femorale of the three internal fixation models were the smallest in the femoral neck system model and the largest in the cannulated compression screw model.(2)Under the single-leg standing load and the sideways fall load,the peak displacement of the proximal femur of the three internal fixation models was all located at the top of the femoral head,and the peak displacement was the smallest in the femoral neck system model and the largest in the cannulated compression screw model.(3)The peak displacement of the three internal fixation models was all located at the top of the internal fixation device under the single-leg standing and sideways fall loading conditions,and the peak displacement values were the smallest in the femoral neck system internal fixation model and the largest in the cannulated compression screw internal fixation model.(4)The internal fixation stress of the three internal fixation models was mainly distributed in the area near the fracture end of the internal fixation device under the single-leg standing and sideways fall loads,and the peak value of internal fixation stress was the smallest in the femoral neck system model and the largest in the cannulated compression screw model.(5)These results suggest that the mechanical stability of the femoral neck system is the best,but there may be a risk of stress shielding of the fracture end and calcar femorale.The stress of the internal fixation device of the femoral neck system is more dispersed,and the risk of internal fixation break is lower.

Objective To compare the clinical efficacy and safety of lumbar intervertebral space implantation with single and two fusion devices in treatment of lumbar instability under intervertebral foraminoscopy.Methods 101 patients(101 vertebral bodies)diagnosed with L4,L5 or S1 segment Ⅰ° to Ⅱ° spondylolisthesis from January 2019 to June 2022 were enrolled and the patients were randomly divided into group A(single fusion device group)and group B(double fusion devices group)by intervertebral foraminoscopy.Among of them,there were 51 cases of vertebral spondylolisthesis in group A and 50 cases of vertebral spondylolisthesis in group B.The patients were followed up at 1 week,1 month,6 months,9 months and 15 months after surgery,and the differences in spondylolisthesis distance,intervertebral space height,intervertebral space fusion,vertebral sedimentation rate,lumbar pain visual analogue scale(VAS)and lumbar Oswerry disability index(ODI)between the two groups were compared.Results All the 101 patients were followed up throughout the whole process,and there were no significant differences in operation time,intraoperative blood loss,and postoperative drainage between the two groups(P＞0.05),and the VAS was(7.32±0.57)in group A and(7.14±0.61)in group B one week after operation,there was no significant difference between the two groups(P＜0.05).At 6 months after surgery,the ODI of group A was(27.44±9.52)%and group B was(21.89±8.46)%,and there was no significant difference between the two groups(P＞0.05).And at 6,9 and 15 months follow-up,the fusion rates of group A were 11.76%,68.63%and 90.20%,respectively,and the fusion rates of group B were 18.00%,80.00%and 96.00%,respectively,and there was significant difference between the two groups(P＜0.05).At 15 months postoperative follow-up,the intervertebral space height of group A(0.45±0.07)cm was significantly smaller than that of group B(0.58±0.06)cm,and the difference was statistically significant(P=0.038).There was no significant difference between group A and group B(0.33±0.18)mm and(0.28±0.21)mm(P=0.079)of spondylolisthesis distance.Conclusion Implantation of the double fusion devices can provide more contact area and better stability,shorten the vertebral fusion time,increase the vertebral fusion rate,and reduce the occurrence of vertebral settlement.It is worthy of clinical promotion and application.

Objective To investigate the association between extensive perivascular space(EPVS)burden in different locations and ischemic stroke(IS),its subtypes,and transient ischemic attack(TIA)through Mendelian randomization(MR)analysis.Methods The summary data from large-scale Genome-wide Association Studies(GWAS)and various MR methods were employed.We applied multivariable MR to mitigate potential confounding factors and conduct sensitivity analyses to enhance result robustness.Subsequently,meta-analysis was utilized to integrate causal relationships between EPVS burden in different locations and IS from various sources.Additionally,reverse MR was employed to observe the impact of various IS types on EPVS burden.Finally,linkage disequilibrium score regression was conducted to assess genetic correlations between exposures and outcomes.Results EPVS burden in both the white matter(OR=1.12,95％CI:1.01-1.25;P=0.04)and basal ganglia(OR=1.57,95％CI:1.30-1.89;P＜0.01)are significant risk factors for IS.EPVS burden in the basal ganglia is also a risk for IS(small-vessel)(OR=4.56,95％CI:2.57-8.27;P=5.95× 10-7).After IS and TIA there seems to be a potential increase in extensive basal ganglia perivascular space burden.Conclusions Extensive white matter perivascular space burden and extensive basal ganglia perivascular space burden may serve as important indicators to predict IS.

Objective:To explore the correlation between clinical and CT imaging features and epidermal growth factor receptor (EGFR) gene mutation in patients with non-small cell lung cancer (NSCLC) and screening of mutation prediction indicators.Methods:A retrospective case-control study was conducted. The clinical data of 178 NSCLC patients who were confirmed by pathology and underwent pre-treatment chest-enhanced CT scan and EGFR gene mutation testing in General Hospital of Ningxia Medical University from January 2015 to December 2019 were retrospectively analyzed. Patients were classified into EGFR mutation-positive and mutation-negative groups based on genetic testing results, and the clinical and CT imaging features were compared between the two groups; the multivariate logistic regression model was used to identify the independent influencing factors for EGFR gene mutation in NSCLC patients.Results:Among 178 NSCLC patients, 115 cases (64.6%) were EGFR gene mutation-positive and 63 cases (35.4%) were mutation-negative. Among the 115 EGFR gene mutation-positive patients, there were 61 cases (53.0%) of exon 19 deletion (19del) mutation, 45 cases (39.1%) of exon 21 L858R mutation, 8 cases (7.0%) of exon 20 mutation, and 1 case (0.9%) of exon 18 mutation. The proportions of female patients [60.0% (69/115) vs. 30.2% (19/63)] and patients with out smoking history [74.8% (86/115) vs. 36.5% (23/63)] in EGFR gene mutation-positive group were higher than those in the mutation-negative group, and the differences were statistically significant (both P < 0.001), while the proportions of patients with different pathological types and clinical stages in the two groups showed no statistically significant differences (both P > 0.05). The median maximum diameter of tumor [ M ( Q1, Q3)] detected by CT in the EGFR gene mutation-positive group was 3.70 (2.90, 4.70) cm, while in the mutation-negative group it was 5.30 (3.40, 6.80) cm, and the difference was statistically significant ( Z = -3.66, P < 0.001). The proportions of patients with air bronchogram [27.8% (32/115) vs. 7.9% (5/63)] and without emphysema [83.5% (96/115) vs. 55.6% (35/63)] in the EGFR gene mutation-positive group were higher than those in the mutation-negative group, and the differences were statistically significant (both P < 0.01). The results of multivariate logistic regression analysis showed that no smoking history (yes vs. no, OR = 0.218, 95% CI: 0.073-0.647), short maximum diameter of tumor detected by CT ( OR = 0.814, 95% CI: 0.676-0.981), air bronchogram (yes vs. no, OR = 5.354, 95% CI: 1.782-16.090), and no emphysema (yes vs. no, OR = 0.289, 95% CI: 0.128-0.653) were independent risk factors for EGFR gene mutation in NSCLC patients (all P < 0.05). Conclusions:Clinical and CT imaging features may relate to EGFR gene mutation status in NSCLC patients, and no smoking history, short maximum diameter of tumor detected by CT, air bronchogram and no emphysema may predict EGFR gene mutation.

Objective To observe the therapeutic effects and mechanism of Xiezhuo Qutong Formula on gout.Methods(1)Animal experiments:A hyperuricemia(HUA)model was established in SD rats using hypoxanthine combined with potassium oxonate,and an acute gouty arthritis(AGA)model was induced by monosodium urate.The efficacy and safety of Xiezhuo Qutong Formula in reducing serum uric acid levels and exerting anti-inflammatory effects were evaluated.(2)Network pharmacology:The main active components and potential targets of Xiezhuo Qutong Formula were collected,along with gout-related disease targets.The intersection of these targets yielded potential therapeutic targets.A protein-protein interaction(PPI)network was constructed,followed by Gene Ontology(GO)functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis.Results Xiezhuo Qutong Formula effectively reduced serum uric acid levels in HUA model rats without observed hepatorenal toxicity.It also decreased joint inflammation index,joint swelling degree,and inflammatory cytokine levels in AGA model rats.Network pharmacology analysis identified 156 overlapping targets between the drug and disease,among which TP53,STAT3,PIK3CA,BCL2,PIK3CD,PIK3CB,JUN,HDAC1,ESR1,and RELA may be key targets of Xiezhuo Qutong Formula in treating gout.Combined with GO and KEGG enrichment analyses,the main involved pathways include PI3K/AKT,JAK2/STAT3,and NF-κB signaling pathways.Conclusion Xiezhuo Qutong Formula exerts uric acid-lowering and anti-inflammatory effects in the treatment of gout,and its potential mechanism involves the PI3K/AKT,JAK2/STAT3,and NF-κB signaling pathways.