Conventional cancer therapies, such as radiotherapy and chemotherapy, often damage normal cells and may induce new tumors. Oncolytic viruses (OVs) selectively target tumor cells while sparing normal cells. Most OVs used in clinical trials have been genetically engineered to enhance their ability to target tumor cells and activate immune responses. To develop a specific OV-based approach for treating cervical cancer, this study constructed an oncolytic adenovirus that delivered a base editor targeting oncogenes to achieve efficient killing of tumor cells through inhibiting tumor growth and directly lysing tumor cells. We utilized the human telomerase reverse transcriptase (TERT) promoter to drive the expression of adenovirus early region 1A (E1A) and successfully constructed the P-hTERT-E1A-GFP vector, which was validated for its activity in cervical cancer cells. Given the critical role of the MYC oncogene in the research of oncology, identifying efficient editing sites for the MYC oncogene is a key step in this study.Three MYC-targeting gRNAs were engineered and co-delivered with ABE8e base editor plasmids into HEK293T cells. Following puromycin selection, Sanger sequencing demonstrated differential editing efficiencies: MYC-1 (43%), MYC-2 (25%), and MYC-3 (35%), identifying MYC-1 as the most efficient editing locus. By constructing the P-ABEs-hTERT-E1A-GFP and P-MYC gRNA-hTERT-E1A-GFP vectors, we successfully packaged the virus and confirmed its specificity and efficacy. The experimental results demonstrate that this novel oncolytic adenovirus effectively inhibits the growth of HeLa cells in vitro, providing new experimental evidence and potential strategies for treating cervical cancer based on the HeLa cell model.
Humans ; Uterine Cervical Neoplasms/pathology* ; Oncolytic Viruses/genetics* ; Female ; HEK293 Cells ; Oncolytic Virotherapy/methods* ; Adenoviridae/genetics* ; Gene Editing/methods* ; Telomerase/genetics* ; Adenovirus E1A Proteins/genetics* ; Genetic Vectors/genetics* ; HeLa Cells

Objective:To evaluate the diagnostic value of native T1 mapping in differentiating Oxford classification (MEST-C) scores in patients with IgA nephropathy.Methods:In this prospective study, patients who underwent both T1 mapping and renal biopsy at the First Medical Center of the Chinese PLA General Hospital between April 2023 and October 2024 were consecutively enrolled. Two radiologists, blinded to clinical and pathological information, measured renal T1 mapping parameters, including cortical T1 (cT1), medullary T1 (mT1), the corticomedullary difference (ΔT1), and the corticomedullary ratio (T1 ratio). Clinical and renal biopsy data based on the Oxford classification from patients with IgA nephropathy were collected. The Oxford classification includes five indicators: Mesangial hypercellularity (M), Endocapillary hypercellularity (E), Segmental glomerulosclerosis or adhesion (S), Tubular atrophy/interstitial fibrosis (T), and Cellular or fibrocellular crescents (C). Spearman correlation analysis was applied to evaluate the associations between MEST-C scores and T1 parameters. The diagnostic performance of T1 parameters for discriminating among scores of the Oxford classification was analyzed using the receiver operating characteristic (ROC) curve.Results:A total of 124 patients with IgA nephropathy were included in this study [66 males, 58 females; age 19-70 years, 39 (30, 51) years]. Except for the E indicator, M, S, T, and C were significantly correlated with renal T1 values ( ρ=0.177-0.414, all P<0.05). cT1 showed the best diagnostic efficacy for the S score, with an area under the curve (AUC) of 0.798, a sensitivity of 68.7%, and a specificity of 88.0%. The best T1 parameter for differentiating the T score was the T1 ratio, with an AUC of 0.687, a sensitivity of 57.9%, and a specificity of 79.1%. Conclusion:Native T1 mapping can be used for the non-invasive assessment of the S and T scores in the Oxford classification of patients with IgA nephropathy.

Objective:To evaluate the effects of chlorinated organophosphate flame retardants (Cl-OPFRs) via respiratory and digestive tract exposure on multiple organs in mice.Methods:A short-term repeated exposure model of tris(2-chloroethyl) phosphate (TCEP), tris(1-chloro-2-propyl) phosphate (TCIPP) and tris(1, 3-dichloro-2-propyl) phosphate (TDCIPP) in mice was established through intratracheal instillation and oral gavage administration. The exposure doses were 0.7, 1 and 2 mg·kg -1·day -1, respectively, with continuous administration for 14 days. The organs of the heart, liver, spleen, lung, kidney, stomach, large intestine, small intestine, bladder and testis were collected and weighed to calculate the organ coefficients. The pathological and histological changes were observed by hematoxylin-eosin staining to quantitatively assess the effects of the three Cl-OPFRs on the various organs by using the pathology score. Results:Analysis of organ coefficients in tracheal drip-treated mice showed that the organ coefficients in the testes of the TCEP, TCIPP and TDCIPP groups were lower than those in the control group ( PTCEP-testis=0.045, PTCIPP-testis=0.012 and PTDCIPP-testis<0.001). The organ coefficients were lower in the lungs and small intestines of the TCEP group ( PTCEP-lung=0.006, PTCEP-small intestine=0.042). The organ coefficients for the stomach and large intestine were higher in the TDCIPP group ( PTDCIPP-stomach=0.014, PTDCIPP-large intestine=0.049). Analyses of gavage-contaminated mice showed that the organ coefficients for liver, stomach and small intestine in the TCEP and TDCIPP groups were higher than those in the control group ( PTCEP-liver=0.007, PTCEP-stomach=0.003, PTCEP-small intestine<0.001, PTDCIPP-liver=0.001, PTDCIPP-stomach=0.004, and PTDCIPP-small intestine<0.001). Histopathological analyses of the organs of tracheal drip dyed mice showed significant pathological damage in the lung tissue of the TCIPP group, mainly in the form of thickening of the interstitium, infiltration of inflammatory cells and alveolar collapse. The results of the analysis of gavage poisoned mice showed that TCIPP exposure could lead to blurring of the red and white medullary boundaries of spleen tissues, destruction of white medullary structures, etc., and induce small intestinal cryptitis. TDCIPP induced significant pathological damage to the liver tissues of mice, which mainly included cytoplasmic washout, infiltration of inflammatory cells, acute inflammation, and other injurious effects. Significant pathological damage to the intestinal tissues of mice was also observed. Conclusions:This study demonstrates that the toxic effects of Cl-OPFRs are significantly associated with exposure routes and compound specificity. Respiratory exposure predominantly induces TCIPP-mediated pulmonary injury, while digestive exposure causes TDCIPP-driven hepatointestinal toxicity. These findings provide preliminary evidence for the toxicity screening of Cl-OPFRs.

Aim To investigate the effect of irisin on endothelial inflammation and atherosclerosis(As)in mice.Methods ApoE-/-mice were randomly divided into control group,As model group,and irisin group(treated with irisin based on the As model),with 10 mice in each group.The carotid tissues were stained using pathological techniques and immunofluorescence.Human aortic endothelial cells(HAEC)were cultured in vitro,treated with irisin,and stimulated with cholesterol crystal(CC).The protein levels of vascular cell adhesion molecule-1(VCAM-1)and intercellular cell adhesion molecule-1(ICAM-1)were then detected by Western blot.The expression of inflammatory cytokines interleukin-1β(IL-1β),interleukin-6(IL-6)and chemokine(C-C motif)ligand 2(CCL2)were detected by RT-qPCR.The ad-hesion of monocytes was assessed using cell adhesion assay.Results The carotid plaque area in the mice of As model group was significantly increased compared with that in control group(P＜0.05).In contrast,the plaque area was re-duced in the irisin group compared with the As model group(P＜0.05).Compared with the control group,the expression of VCAM-1,the number of CD68+macrophages,and the deposition of CC were increased in the carotid arteries of the As model group(P＜0.05),while irisin could reduce the expression of VCAM-1,the number of CD68+macrophages,and the deposition of CC(P＜0.05).At the in vitro level,the expression of VCAM-1 and ICAM-1,as well as the adhesion of monocytes in CC-stimulated HAEC,were increased(P＜0.05).However,irisin could inhibit the increased expression of VCAM-1 and ICAM-1(P＜0.05),as well as the adhesion of monocytes induced by CC(P＜0.05).The mRNA levels of IL-1β,IL-6 and CCL2 in HAEC of CC stimulated group were increased(P＜0.05),while irisin could inhibit the mRNA expressions of IL-1β,IL-6 and CCL2 induced by CC(P＜0.05).Conclusion Irisin can inhibit vascular inflamma-tion,thereby reducing the occurrence and progression of atherosclerosis.

Metering unit is important component of medical service items and also serve as significant bases for scientifically estimating the costs of medical services.National Technical Specification of Medical Service Items(2023 Edition)has set up elements such as"metering unit"and"adjustment coefficient for resource consumption in special circumstances",which clearly stipulate the metering units of each medical service item and the multiples for discretionary increases or decreases in the overall resource consumption when special circumstances occur.These two elements are important references for the cost accounting in medical institutions.The project team has elaborated on the definitions,establishment principles and special circumstances of these two types of elements,namely"metering units"and"adjustment coefficient for resource consumption in special circumstances",providing references for the application of relevant governments and healthcare institutions.

The relative value of human resource consumption is a relative value scale of the workload and difficulty of medical personnel needed to provide medical service items.Based on"basic human resource consumption and time consumption","technical difficulty"and"risk level"of the medical service items,it is comprehensively calculated by"relative value indicator standardization","relative value indicator weight setting","relative value indicator calculation model construction",and"relative value indicator expert verification".By formulating the relative value of human resource consumption,it could be used in rationalizing the relative value scale of medical service items,promoting cost accounting in hospitals,and establishing a scientific and reasonable performance evaluation system as technical tools.

The assignment of the"item code"element in National Technical Specification of Medical Service Items(Version 2023)continues the classification framework and coding rules of National Specification of Medical Service Price Items(Version 2012).The project team experts are members of the WHO-Family of International Classifications(WHO-FIC)China Classification,Terminology and Standards Collaborating Center.The design concept of project coding is the same as that of the International Classification of Health Interventions(ICHI).Both adopt an alphanumeric characters hybrid coding classification method based on multi-axis classification,clarified the definition and scope of these four key classification axis which including anatomy system,anatomical site,surgical procedure and surgical approach,and optimize and update item property,sub-classifications and coding.Unified medical service item code can make contribution to the comparability of statistics related to health care items across regions,promote cross-regional healthcare and healthcare insurance settlement at the national level;improve its refined management level in hospital,and play an important role in performance management and data statistical analysis.

Objective:To explore the effects of neutrophil extracellular traps (NET) on the proliferation and migration of breast cancer cells and the related mechanisms.Methods:The peripheral blood neutrophils were separated using density gradient centrifugation, 100 nmol/L phorbol 12-myristate 13-acetate (PMA) was used to stimulate the formation of NET. Breast cancer cell line MDA-MB-231 was selected and treated with NET (NET group) or deoxyribonucleaseⅠ (DNase Ⅰ) digested NET (NET+DNase Ⅰ group),normal cultured MDA-MB-231 cells were used as the control group. In MDA-MB-231 cells of each group; cell proliferation ability was detected using CCK-8 assay; cell migration was detected by cell scratch assay and Transwell assay; real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the relative expressions of frizzled homolog 10 (FZD10), cyclin D1, E-cadherin and Vimentin immunofluorescence assay was performed to detect the formation of NET and the expressions of E-cadherin and Vimentin; Western blotting was used to detected the expression of FZD10-Wnt-β-catenin signaling pathway-related proteins.Results:Neutrophils were stimulated with PMA for 4 hours, and immunofluorescence assay results showed that NET exhibited fibrous reticular structure with high expression of citrullinated histone (citH3) and myeloperoxidase (MPO). The CCK-8 assay results showed that the absorbance values of cells in the control group, NET group and NET+DNase Ⅰ group were 1.25±0.06, 2.14±0.15 and 1.47±0.11, respectively, and the difference was statistically significant ( F = 80.60, P < 0.001). The results of scratch assay displayed that the percentage of wound closure in the control group, NET group and NET+DNase Ⅰ group were (36.2±1.3)%, (67.2±2.0)% and (46.3±3.2)%, respectively, and the difference was statistically significant ( F = 140.50, P < 0.001). Transwell assay indicated that the number of migrating cells in the control group, NET group and NET+DNase Ⅰ group were 317±18, 512±23 and 356±23, respectively, and the difference was statistically significant ( F = 75.39, P < 0.001). qRT-PCR assay revealed that the relative expressions of FZD10, cyclin D1 and Vimentin mRNA in the NET group were higher than those in the control group, while the relative expression of E-cadherin mRNA was lower than that in the control group, and the differences were statistically significant (all P < 0.001); the relative expressions of FZD10, cyclin D1 and Vimentin mRNA in the NET+DNase Ⅰ group were lower than those in the NET group, while the relative expression of E-cadherin mRNA was higher than that in the NET group, and the differences were statistically significant (all P < 0.001). Immunofluorescence assay indicated that the number of cells expressing Vimentin in the control group, NET group and NET+DNase Ⅰ group were 35±6, 86±13 and 51±6, respectively, and the difference was statistically significant ( F = 24.65, P = 0.001); the number of cells expressing E-cadherin were 31±4, 11±3 and 24±2, respectively, and the difference was statistically significant ( F = 38.36, P < 0.001). Western blotting results demonstrated that the relative expressions of FZD10, β-catenin and Vimentin proteins in the NET group were higher than those in the control group, while the relative expression of E-cadherin protein was lower than that in the control group; the relative expressions of FZD10, β-catenin and Vimentin proteins in the NET+DNase Ⅰ group were lower than those in the NET group, while the relative expression of E-cadherin protein was higher than that in the NET group. Conclusions:NET may promote the proliferation and migration of breast cancer cells by upregulating the expression of FZD10 and subsequently activating the Wnt-β-catenin signal pathway.

OBJECTIIVE:The positive role of exercise intervention in the prevention and treatment of sarcopenia has received widespread attention,but the optimal exercise dose for elderly sarcopenic patients still needs to be further determined.The article explored the dose-effect relationship between various elements of exercise prescription and the improvement of muscle mass,muscle strength and physical function in community-dwelling elderly patients with sarcopenia,aiming to provide scientific support for the development of exercise prescription for community-dwelling elderly patients with sarcopenia.METHODS:Literature published from the inception to October 9,2024 in PubMed,EMBASE,Scopus,Web of Science,CNKI,WanFang,VIP,and CBMdisc databases was systematically searched.Meta-analysis was performed using RevMan 5.3 software.Standardized mean difference(SMD)and its 95％CI were used as effect statistics.RESULTS:(1)A total of 11 randomized controlled trials were included,with 348 in the trial group and 304 in the control group.(2)Meta-analysis results showed that exercise improved appendicular skeletal muscle mass index,grip strength,and walking speed in elderly patients with sarcopenia(SMDs 0.46,0.63,0.67,P＜0.05).(3)When the frequency of exercise was 2-3 days/week,appendicular skeletal muscle mass index(SMD=0.57,95％CI:0.28-0.86,P＜0.001),grip strength(SMD=0.70,95％CI:0.37-1.02,P＜0.001),and walking speed(SMD=0.69,95％CI:0.20-1.18,P=0.006)were effectively improved in elderly patients with sarcopenia.(4)When the duration of exercise was 25-60 minutes per session,appendicular skeletal muscle mass index(SMD=0.28,95％CI:0.07-0.50,P=0.01),grip strength(SMD=0.37,95％CI:0.16-0.59,P＜0.001),and walking speed(SMD=0.39,95％CI:0.06-0.73,P=0.02)were effectively improved in elderly patients with sarcopenia.(5)When the exercise intensity was moderate,appendicular skeletal muscle mass index(SMD=0.69,95％CI:0.35-1.03,P＜0.001),and grip strength(SMD=0.36,95％CI:0.09-0.64,P=0.009),and walking speed(SMD=0.91,95％CI:0.34-1.47,P=0.002)were effectively improved in elderly patients with sarcopenia.(6)When the dose of exercise cycle was 8-12 weeks,appendicular skeletal muscle mass index(SMD=0.42,95％CI:0.20-0.64,P＜0.001),grip strength(SMD=0.45,95％CI:0.26-0.64,P＜0.001),and walking speed(SMD=0.76,95％CI:0.27-1.25,P=0.002)were effectively improved in elderly patients with sarcopenia.CONCLUSION:Active,regular exercise can improve muscle health in older adults with sarcopenia.It is recommended that older patients with sarcopenia exercise at least 2 to 3 days per week,25 to 60 minutes each time,lasting for 8 to 12 weeks of moderate intensity exercise to improve muscle health.

Objective To observe the value of T2 mapping for quantitatively evaluating the changes of junctional zone and outer myometrium caused by endometrial fibrosis.Methods A total of 73 infertility patients with endometrial fibrosis confirmed by hysteroscopy(disease group)and 33 healthy women of childbearing age(control group)were prospectively enrolled,and MR examinations were performed at the late proliferative phase of endometrium.The thickness and T2 value of junctional zone,T2 value of outer myometrium on anterior,posterior and fundus wall of midsagittal corpus uteri were measured,and the mean value of the above measurements on the three walls were calculated.Receiver operating characteristic curves were constructed,the areas under the curves(AUC)were calculated to explore the efficacy of those with significant difference among the mean thickness and the mean T2 value of junctional zone,the mean T2 value of outer myometrium and their combination for evaluating endometrial fibrosis.Results The thickness and T2 value of anterior wall,posterior wall,fundus wall and the mean junctional zone in disease group were all significantly higher than those in control group(all P＜0.001).No significant difference of T2 value of anterior wall,posterior wall,fundus wall nor the mean outer myometrium was found between groups(all P＞0.05).The mean thickness and the mean T2 value of junctional zone and their combination could be used to effectively evaluate endometrial fibrosis,with AUC of 0.839,0.822 and 0.922,respectively,and their combination had the best performance(both P＜0.01).Conclusion T2 mapping could be used to quantitatively evaluate the injury of junctional zone caused by endometrial fibrosis.