[摘 要] 目的：探讨RNA结合蛋白15（RBM15）通过三磷酸腺苷酶家族蛋白3A（ATAD3A）调控Wnt/β-catenin通路对宫颈癌细胞恶性生物学行为的影响。方法：利用TCGA数据库分析宫颈癌组织中RBM15 mRNA表达水平及其与患者预后的关系。收集2024年1月至10月期间在赣州市人民医院手术切除的32例宫颈癌组织及癌旁组织标本，以及宫颈癌细胞HeLa、MS-751、C-33A和SiHa，通过免疫组化、WB法检测宫颈癌组织和细胞中RBM15的表达水平。利用SRAMP在线数据库筛选ATAD3A mRNA的m6A修饰位点。通过RNA免疫沉淀实验、RNA衰变实验及挽救实验鉴定RBM15与ATAD3A mRNA的相互作用。采用RNA干扰技术和病毒感染技术，在宫颈癌HeLa和SiHa细胞敲低或过表达RBM15和ATAD3A，qPCR和WB法检测mRNA和蛋白表达，CCK-8法、划痕实验和Transwell实验检测各组细胞的增殖、迁移和侵袭能力。结果：宫颈癌组织中RBM15 mRNA和蛋白阳性率均显著高于癌旁组织（均P < 0.001）。宫颈癌HeLa、MS-751、C-33A和SiHa细胞RBM15蛋白表达水平显著高于正常宫颈细胞Ect1/E6E7（均P < 0.01）。RBM15 mRNA高表达组患者5年无进展生存率低于低表达组患者（P < 0.01）。宫颈癌组织中ATAD3A的表达水平显著高于癌旁组织（P < 0.001），RBM15 mRNA与ATAD3A mRNA呈正相关（r = 0.601，P < 0.05）。ATAD3A mRNA的501、5 312、12 137位点存在高可信度的m6A修饰位点。HeLa、SiHa细胞中过表达RBM15后，ATAD3A mRNA和蛋白表达升高，而敲低RBM15后，ATAD3A mRNA和蛋白表达降低（均P < 0.001）。RNA免疫沉淀实验显示，与IgG组相比，RBM15抗体的免疫沉淀中ATAD3A mRNA显著富集（均P < 0.001）。MeRIP-qPCR实验显示，ATAD3A mRNA 501、5 312、12 137位点均存在明显的m6A甲基化富集（均P < 0.001）。RNA衰变实验显示，敲低RBM15能降低HeLa、SiHa细胞ATAD3A mRNA的半衰期和稳定性（均P < 0.001）。敲低HeLa、SiHa细胞RBM15的表达，显著抑制癌细胞的增殖、迁移及侵袭，Wnt/β-catenin通路相关蛋白Wnt3、β-catenin、vimentin表达均显著降低（均P < 0.001）；而过表达ATAD3A可完全逆转上述抑制作用（均P < 0.001）。结论：RBM15通过m6A修饰ATAD3A mRNA调控Wnt/β-catenin信号通路，从而促进宫颈癌细胞增殖、迁移及侵袭。

More than 70%of tumor patients require radiotherapy.Medical electron linear accelerators are important high-end radiotherapy equipment for tumor radiotherapy.With the application of artificial intelligence technology in medical electron linear accelerator,radiotherapy has evolved from ordinary radiotherapy to today's intelligent radiotherapy.This study introduces the development history,working principles and system composition of medical electron linear accelerators.It outlines the key technologies for improving the performance of medical linear electron accelerators,including beam control,multi-leaf collimator,guiding technology and dose evaluation.It also looks forward to the development trend of major radiotherapy technologies,such as biological guided radiotherapy,FLASH radiotherapy and intelligent radiotherapy,which provides references for the development of medical electron linear accelerators.

Introduction::The triglyceride to high-density lipoprotein cholesterol (TG/HDL-C) ratio, a novel biomarker for metabolic syndrome (MetS), has been validated in the general population as being significantly correlated with cardiovascular disease (CVD) risk. However, its capabilities to predict CVD in people living with human immunodeficiency virus (HIV; PLWH) remain underexplored.Methods::We conducted a retrospective cohort study of 16,081 PLWH who initiated antiretroviral therapy (ART) at the Third People’s Hospital of Shenzhen (China) from 2005 to 2022. The baseline TG/HDL-C ratio was calculated as TG (mmol/L) divided by HDL-C (mmol/L). We employed a multivariate Cox proportional hazards model to assess the association between the TG/HDL-C ratio and CVD occurrence, using Kaplan-Meier curves and log-rank tests to compare survival distributions. The increase in prediction risk upon the addition of the biomarker to the conventional risk model was examined through the assessment of changes in net reclassification improvement (NRI) and integrated discrimination improvement (IDI). Nonlinear relationships were investigated using a restricted cubic spline plot, complemented by a two-piecewise Cox proportional hazards model to analyze threshold effects.Results::At the median follow-up of 70 months, 213 PLWH developed CVD. Kaplan-Meier curves demonstrated a significant association between the increased risk of CVD and a higher TG/HDL-C ratio (log-rank P <0.001). The multivariate-adjusted Cox proportional hazards regression model indicated that the CVD hazard ratios (HR) (95% confidence intervals [95% CIs]) for Q2, Q3, and Q4 versus Q1 of the TG/HDL-C ratio were 2.07 (1.24, 3.45), 2.17 (1.32, 3.57), and 2.20 (1.35, 3.58), respectively ( P <0.05). The consideration of the TG/HDL-C ratio in the model, which included all significant factors for CVD incidence, improved the predictive risk, as indicated by the reclassification metrics (NRI 16.43%, 95% CI 3.35%-29.52%, P = 0.014). The restriction cubic spline plot demonstrated an upward trend between the TG/HDL-C ratio and the CVD occurrence ( P for nonlinear association = 0.027, P for overall significance = 0.009), with the threshold at 1.013. Significantly positive correlations between the TG/HDL-C ratio and CVD were observed below the TG/HDL-C ratio threshold with HR 5.88 (95% CI 1.58-21.88, P = 0.008), but not above the threshold with HR 1.01 (95% CI 0.88-1.15, P = 0.880). Conclusion::Our study confirms the effectiveness of the TG/HDL-C ratio as a predictor of CVD risk in PLWH, which demonstrates a significant nonlinear association. These findings indicate the potential of the TG/HDL-C ratio in facilitating early prevention and treatment strategies for CVD among PLWH.

Objective To investigate the relationship between systemic immune-inflammation index (SII)and lower extremity vascular disease in patients with type 2 diabetes mellitus (T2DM).Methods A cross-sectional study was conducted on 390 T2DM patients admitted in our department from January 2013 to January 2024.According to the diagnostic criteria for lower extremity vascular disease in T2DM patients,they were divided into a lower extremity vascular disease group (n=158)and a control group (n=232).General data and results of laboratory tests were compared between the 2 groups.Spearman correlation analysis was used to identify the related factors for lower extremity vascular diseases in T2DM patients.The correlation between SII and lower extremity vascular diseases in T2DM patients was analyzed using the Row Mean Scores and Cochran-Armitage Trend analysis.Multivariate logistic regression analysis was applied to identify the risk factors for lower limb vascular lesions in T2DM patients.Receiver operating characteristic (ROC)curve was plotted to evaluate the diagnostic efficacy of SII for lower extremity vascular disease in the patients.Results Compared with T2DMpatients without lower extremity vascular disease,those with lower extremity vascular disease were older,had higher levels of total cholesterol (TC),low-density lipoprotein cholesterol (LDL-C),SII,larger proportion of carotid vascular lesions,and increased proportion of no-taking statins.The lower extremity vascular disease in T2DM patients was positively correlated with SII/100 (r=0.429,P<0.001),age (r=0.517,P<0.001),TC (r=0.161,P=0.001),LDL-C (r=0.117,P=0.021),carotid artery lesions (r=0.101,P=0.047),no-taking statins (r=0.266,P<0.001).Logistic regression analysis showed that SII,age,LDL-C,and no-taking statins were the risk factors for lower extremity vascular lesions in T2DM patients (P<0.01).The area under the curve (AUC)value of SII combined with age,LDL-C,and no-taking statins in predicting lower extremity vascular disease in T2DM patients was 0.896.Conclusion SII is not only a risk factor,but also a simple marker for lower extremity vascular disease in T2DM patients,suggesting that inflammatory response plays an important role in the occurrence and development of lower extremity vascular disease in T2DM.

In order to search for coumarin-based anti-platelet aggregation compounds with high efficacy and good druggability,twenty-five 3-acetyl-7-hydroxy-coumarin oxime derivatives(6a-6y)were synthesized via Vilsmeier-Haack reaction,Knoevenagel reaction,Williamson reaction,electrophilic substitution reaction and oximation reaction from resorcinol.Their structures were confirmed by HRMS and 1H NMR spectra.The anti-platelet aggregation activity of the target compounds was evaluated using Born's turbidimetric method.The results revealed that most of them could significantly inhibit platelet aggregation induced by adenosine diphosphate(ADP),collagen,arachidonic acid(AA)and thrombin.Among them,the target compounds 6a and 6b not only had strong inhibitory activity on platelet aggregation induced by the four inducers,but also exhibited good water solubility(3.46 mg/mL and 3.85 mg/mL,respectively)and lipid-water partition coefficient(2.56 and 2.85,respectively)and were expected to become a preclinical candidate compound with multi-target action against platelet aggregation.

Objective To explore the viral pathogen spectrum characteristics of acute intestinal infection in Hanzhong from 2019 to 2022. Methods Fecal samples from patients with acute intestinal infection in the outpatient clinic of 3201 Hospital from January 2019 to December 2022 were collected. Common enteroviruses such as enterovirus 71 (EV-A71), coxsackievirus 16 (CV-A16), CV-A10, CV-A6, CV-A2, CV-A4, and CV-B3 were detected and analyzed by real-time fluorescence quantitative PCR. Results A total of 5 194 fecal samples were collected, and the positive rate of nucleic acid detection was 23.95%. In terms of the enteroviruses, the highest detection rate was 9.82% for EV-A71, followed by 4.58% for CV-A16 and 3.37% for CV-A6. The positive detection rate of common enteroviruses showed statistical difference among different age groups (P<0.05), with the highest detection rate of 41.49% in 0-4 years old group. There was no significant difference in the positive virus detection rate between different genders (P>0.05). EV-A71 infection showed no seasonal characteristics, whereas the detection of CV-A16 and CV-A6 infections was concentrated in summer and autumn. There were 106 cases of mixed infection, and the prevalence rate was 2.04%, with EV-A71 and CV-A6 mixed infections accounting for the majority of cases. Conclusion The main pathogens of acute intestinal infections in the Hanzhong area from 2019 to 2022 are EV-A71, CV-A16, and CV-A6. It is necessary to strengthen the monitoring of acute intestinal infections in children aged 4 years and below.

Objective To study the role of microRNA(miR)-483-3p in reducing myocardial fibrosis in rats,and explore the relationship between its mechanism and autophagy.Methods A total of 24 male SD rats were randomly divided into sham operation group,model group,blank transfec-tion group and high expression group,with 6 rats in each group.The blank transfection group and the high-expression group were pretreated with a single injection of adeno-associated virus(AAV)-blank transfection and AAV-miR-483-3p(5×1011 vg)in the tail vein,respectively.In 14 d later,the sham group was injected with 2.5 ml/(kg·d)normal saline for 14 d,and rat model of myocardial fibrosis was established by 2 mg/ml isoproterenol[2.5 ml/(kg·d)]injection through tail vein for 14 consecutive days.Myocardial pathological damage,severity of myocardial fibrosis,and expression levels of collagen-Ⅰ,microtubule-associated protein light chain 3(LC3),autoph-agy-related protein 5(Atg5)and autophagy degradation substrate(P62)in cardiomyocytes were evaluated and measured.Results Compared with the sham operation group,the model group had obviously larger myocardial fibrosis area,higher positive expression of Collagen-Ⅰ,and increased protein levels of Atg5 and LC3-Ⅱ/LC3-Ⅰ,and decreased expression level of P62 protein(P＜0.05).The myocardial fibrosis area,positive expression of Collagen-Ⅰ,the expression levels of Atg5 and LC3-Ⅱ/LC3-Ⅰ protein[(13.64±1.51)％vs(27.47±1.55)％,(13.48±3.07)％vs(30.91±2.45)％,0.98±0.17 vs 1.24±0.28,0.66±0.05 vs 1.26±0.09,P＜0.05]were significant-ly decreased,and the expression level of P62 was notably increased(0.91±0.11 vs 0.74±0.06,P＜0.05)in the high expression group than the model group.Conclusion MiR-483-3p attenuates myocardial fibrosis in rats,and the mechanism may be related to the inhibition of cardiomyocyte autophagy.

Advanced radiotherapy technology enables the dose to more accurately conform to the tumor target area of the patient, providing accurate treatment for the patient, but the gradient of the patient's radiation dose at the tumor edge is getting larger, which putting forward higher requirements for radiotherapy dose verification. The dose verification system software KylinRay-Dose4D can verify the patient's pre-treatment plan and the in vivo/on-line dose during the patient's treatment, providing important reference for the physicist to modify the radiotherapy plan and ensuring that the patient receives accurate treatment. This study introduces the overall design and key technologies of KylinRay-Dose4D, and tests the pre-treatment plan dose checking calculation and 2D/3D dose verification through clinical cases. The test results showed that the 2D/3D gamma pass rate (3 mm/3%) of KylinRay-Dose4D reconstructed dose compared with TPS plan dose and measured dose is larger than 95%, which indicating that the reconstructed dose of KylinRay-Dose4D meets the requirement of clinical application.
Humans ; Radiotherapy Dosage ; Radiotherapy Planning, Computer-Assisted/methods* ; Radiotherapy, Intensity-Modulated/methods* ; Software ; Neoplasms ; Phantoms, Imaging ; Radiometry/methods*

Background::Understanding the characteristics of newly diagnosed primary human deficiency virus-1 (HIV-1) infection in the context of the post-antiretroviral therapy era and HIV drug prophylaxis is essential for achieving the new target of 95-95-95-95 by 2025. This study reported the characteristics of newly diagnosed primary HIV-1 infection in Shenzhen.Methods::This is a real-world retrospective study. Eighty-seven newly diagnosed primary HIV-1-infected patients were recruited from January 2021 to March 2022 at the Third People’s Hospital of Shenzhen. Demographic, epidemiological, diagnostic, drug resistance, and medical data were described and analyzed.Results::Overall, 96.6% (84/87) of the newly identified primary HIV-1-infected patients were male, including 88.5% (77/87) men have sex with men (MSM), with a median age of 29.0 years (Q 1-Q 3: 24.0-34.0 years); of these, 85.1% (74/87) reported high-risk sexual behaviors with casual partners. The rate of condom usage was only 28.7% (25/87). The overall rate of pre-exposure prophylaxis (PrEP) and post-exposure prophylaxis (PEP) was 8.0% (7/87, including 4 PrEP and 3 PEP cases) around the potential exposure, although 41.4% of the patients had prior awareness of such interventions. Moreover, only 19.5% (17/87) had previously used PrEP or PEP. Of those, 58.8% (10/17) of the patients obtained drugs from the internet, and only 35.3% (6/17) reported good compliance. A total of 54.0% (47/87) of subjects were diagnosed by the HIV nucleic acid test. Acute retroviral syndrome appeared in 54.0% (47/87) of patients. The prevalence of transmitted drug resistance (TDR) mutation was 33.9% (19/56), including 6 (10.7%) against nucleoside reverse transcriptase inhibitor (NRTI) plus non-nucleoside reverse transcriptase inhibitor (NNRTI), 8 (14.3%) against NNRTI, and 5 (8.9%) against protease inhibitor (PI) only. Conclusions::Owing to the low utilization rate and incorrect usage of PrEP and PEP, massive efforts are needed to promote HIV-preventive strategies in the MSM population. The extremely high prevalence of TDR mutation in this population implies the need for future pretreatment drug resistance surveillance.