mRNA-driven CAR-T cell therapy has emerged as an important direction in non-viral CAR-T development because it avoids genomic integration, enables the temporal regulation of CAR expression, and facilitates the in vivo generation of CAR-T cells. The key challenge is no longer simply whether CAR expression can be achieved, but how to balance a sufficient duration of effector activity with reversibility and efficient in vivo delivery. Conventional linear mRNA benefits from well-established manufacturing processes but is limited by its relatively short expression period, whereas circular mRNA can extend the expression window. Compared with conventional circular mRNA, scarless circular mRNA minimizes exogenous scar sequences introduced by classical circularization systems, thereby reducing uncertainty in molecular structure and simplifying quality control. These features may make it particularly suitable for delivering long coding sequences such as CAR constructs. Major challenges for future clinical translation include T cell-specific delivery, precise control of the in vivo expression duration, the safety of repeated administration, and the development of scalable manufacturing processes and quality standards. This review summarizes recent advances in mRNA-engineered CAR-T cell therapy, focusing on the construction, functional value, and translational potential of scarless circular mRNA.

Objective To establish a droplet digital PCR(ddPCR)method for detecting the Septin9 gene methylation in colorectal cancer(CRC)and evaluate its performance.Methods Specific primers and probes were designed for the methylation of Septin9 gene.The reaction conditions of ddPCR for Septin9 gene methyl-ation detection were optimized,including primer and probe concentrations,annealing temperature and cycle number.A ddPCR method to detect the methylation of Septin9 gene was established.The linearity,sensitivi-ty,specificity,precision,accuracy and clinical diagnostic accuracy of the established method were evaluated by detecting methylation reference materials and clinical samples.Results The optimal concentrations of primers and probes were 500 nmol/L and 200 nmol/L,respectively.The optimal annealing temperature was 56℃and the optimal number of cycles was 45.The linearity test showed good linearity in the range of 5-104 copy/re-action,and the preset detection limit of Septin9 gene methylation concentration detected by ddPCR was 0.422%,which could specifically identify Septin9 gene methylation positive control.In precision evaluation,the coefficients of variation of high concentration and low concentration reference materials were 1.340 0%and 3.330 0%,which met the relevant requirements.The results of accuracy test showed that the positive co-incidence rate and negative coincidence rate of 10 repeated tests of methylation quality control samples of 3 batches were both 100%.The results of clinical samples showed that the sensitivity and specificity of ddPCR detection of Septin9 gene methylation in CRC group were 82.61%(95%CI 66.10%—99.10%)and 78.38%(95%CI 65.20%—91.60%),respectively.The area under the curve was 0.881 3(95%CI 0.784-0.978 6).Conclusion The ddPCR method for detection of Septin9 gene methylation in CRC has high sensitivity,speci-ficity,precision,accuracy and clinical diagnostic accuracy.It can provide a reliable technical means for early di-agnosis,monitoring and treatment of CRC.

Objective:To evaluate the feasibility of using the domestic ArcherQA system for fast and simplified independent verification of CyberKnife (CK) stereotactic radiotherapy (SRT) plans.Methods:SRT plans of 57 patients treated with CK at Tianjin Medical University Cancer Institute and Hospital from August 2021 to August 2022 were retrospectively analyzed, including 15 intracranial, 30 pulmonary, and 12 abdominal tumors cases. Point-dose and planar-dose verifications were performed using an ionization chamber and radiochromic films embedded in a homogeneous phantom, and the results were compared with those calculated by the treatment planning system (TPS). The localization CT images and corresponding SRT plans were imported into the ArcherQA system for independent dose verification and analysis. The correlation between ArcherQA results and phantom measurements was analyzed, with comparisons of target mean dose differences and γ pass rates.Results:Phantom measurement results showed, the measured point-dose differences for intracranial, lung, and abdominal plans were -0.94% ± 3.22%, 1.92% ± 2.05%, and 2.12% ± 0.77%, respectively. The mean dose differences in target dose calculation between ArcherQA and TPS: intracranial in the gross tumor volume (GTV) regions were 0.34% ± 2.21%, lung tumor GTV were -2.47% ± 2.46%, and abdominal tumor GTV were 0.80% ± 2.61%, respectively. Among them, the abdominal GTV region showed the highest correlation between ArcherQA and measured results ( r=0.78). The average two-dimensional γ pass rates (2 mm/2%, threshold=10%) measured using phantom films were 95.92% ± 2.35% for intracranial, 95.70% ± 2.74% for lung, and 96.74% ± 3.41% for abdominal tumors plans, respectively. The three-dimensional ArcherQA results showed comparable γ pass rates (1 mm/2%, threshold=10%) for lung and abdominal GTV and PTV regions, with similar medians and data dispersion to film measurements. Conclusions:The ArcherQA system enables rapid and efficient independent dose verification of CK SRT plans without the need for additional hardware. The verification results show good correlation with phantom measurements, supporting its potential as an auxiliary quality assurance tool in clinical CK SRT implementation.

Objective:To evaluate the feasibility of using the domestic ArcherQA system for fast and simplified independent verification of CyberKnife (CK) stereotactic radiotherapy (SRT) plans.Methods:SRT plans of 57 patients treated with CK at Tianjin Medical University Cancer Institute and Hospital from August 2021 to August 2022 were retrospectively analyzed, including 15 intracranial, 30 pulmonary, and 12 abdominal tumors cases. Point-dose and planar-dose verifications were performed using an ionization chamber and radiochromic films embedded in a homogeneous phantom, and the results were compared with those calculated by the treatment planning system (TPS). The localization CT images and corresponding SRT plans were imported into the ArcherQA system for independent dose verification and analysis. The correlation between ArcherQA results and phantom measurements was analyzed, with comparisons of target mean dose differences and γ pass rates.Results:Phantom measurement results showed, the measured point-dose differences for intracranial, lung, and abdominal plans were -0.94% ± 3.22%, 1.92% ± 2.05%, and 2.12% ± 0.77%, respectively. The mean dose differences in target dose calculation between ArcherQA and TPS: intracranial in the gross tumor volume (GTV) regions were 0.34% ± 2.21%, lung tumor GTV were -2.47% ± 2.46%, and abdominal tumor GTV were 0.80% ± 2.61%, respectively. Among them, the abdominal GTV region showed the highest correlation between ArcherQA and measured results ( r=0.78). The average two-dimensional γ pass rates (2 mm/2%, threshold=10%) measured using phantom films were 95.92% ± 2.35% for intracranial, 95.70% ± 2.74% for lung, and 96.74% ± 3.41% for abdominal tumors plans, respectively. The three-dimensional ArcherQA results showed comparable γ pass rates (1 mm/2%, threshold=10%) for lung and abdominal GTV and PTV regions, with similar medians and data dispersion to film measurements. Conclusions:The ArcherQA system enables rapid and efficient independent dose verification of CK SRT plans without the need for additional hardware. The verification results show good correlation with phantom measurements, supporting its potential as an auxiliary quality assurance tool in clinical CK SRT implementation.

Objective:The quality of domestic diagnostic accuracy research was evaluated to explore the quality level of domestic diagnostic accuracy research based on Standards for Reporting of Diagnostic Accuracy 2015 (STARD2015).Methods:The Chinese core journals of CNKI and Wanfang Database were searched, and research literature on diagnostic accuracy published from 2017 to 2022 were collected. The main search terms are diagnostic test, sensitivity, specificity, receiver operating characteristic (ROC) curve, etc. The literature was selected according to the inclusion criteria, and the quality of the included literature was evaluated independently by two reviewers, and the conformity rate of the STARD2015 report and the STARD2015 article was calculated. The report quality of the literature was divided into three levels (low, medium and high) according to the conformity rate of the STARD2015 report. The proportion of literature at each level and the proportion of literature at medium and high level in each year were counted. According to quality analysis on the contents of the included articles, articles were divided into standardized reports and unstandardized reports. Intragroup correlation coefficient ( ICC) was used to analyze the consistency of two reviewers. The trend Chi-square test was used to analyze the trend of the proportion of medium and high level literature in each year. One-way analysis of variance was used to compare the coincidence rates evaluated by STARD2015 for each year. Results:A total of 6 771 studies on diagnostic accuracy published from 2017 to 2022 were included. The compliance rate sccording to STARD2015 was 39.56%±4.90%, and the reported compliance rate ranged from 17.65% to 64.71% (the number of reported items ranged from 6 to 22), and 93.53% (6 333/6 771) literatures were in the middle level. Compliance rate of STARD2015 reports varied significantly among different years ( F=25.023, P<0.01), and the compliance rate of 2021 was significantly higher than that of other years ( P<0.01). The proportion of medium and high level literatures according to STARD2015 showed an increasing trend ( χ 2=14.099, P<0.01). The reporting situation of each item varied significantly, and the conformity rate of items raned from 0 to 100%. According to report item, non-standard report rate was 10.34% (569/5 503) for item 6, 4.15% (277/6 677) for item 8, 21.84% (1 447/6 626) for item 20, 66.67% (24/36) for item 22, and 26.03% (877/3 369) for item 26. Conclusions:The overall report quality of published domestic literature on diagnostic accuracy from 2017 to 2022 is at a medium level according to STARD2015, and the reports conformity rate of each item vary significantly, indicating significant knowledge gap on STARD2015 among domestic researchers. The promotion of STARD2015 needs to be strengthened.

Objective The purpose of this investigation was to study the effects of the Sildenafil citrate on mice hearing.Methods Seven-week-old adult male Kunming mice were used.The mice were randomly divided into four groups with 10 mice in each group.Sildenafil groups were orally administered daily with sildenafil [0.1 mg/(kg · d),1 mg/(kg · d),10 mg/(kg · d)] and control group was orally administered with normal saline.Then mice were tested for auditory brainstem response(ABR) to observe the changes of ABR's thresholds at before administration and 1,5,10,15,20 day afterwards.The mice basilar membrane samples were studied by immunofluorescent labeling.High performance liquid chromatography was used for determination the concentration of sidenafil in endolymph of mice cochlea.Statistical analysis was performed using SPSS 13.0.Results After 30 min following administration,the Sildenafil in endolymph of mice cochlear could be assayed by high performance liquid chromatography,and it was doserelated.Sildenafil increased the hearing thresholds with the time of administration.Hearing thresholds increased significantly in the sidenafil group at 20 d compared to the control group (P ＜ 0.05).After administered high dose of Sildenafil,on the 20th day,the ABR thresholds average threshold was (60.0 ± 10.0) dBnHL,and the control group was (14.5 ± 6.0) dBnHL.Hair cells damages in the base ring of cochlea could be observed in experimental group in a concentration-dependent manner.Conclusion Sidenafil can pass through blood-labyrinth barrier to the inner ear,and doses of sildenafil administration can induce hearing impairment in mice.