[摘 要] 自2017年以来，已有12款嵌合抗原受体基因修饰T淋巴细胞（CAR-T细胞）产品相继被批准用于血液系统恶性肿瘤的治疗，包括复发性/难治性急性B淋巴细胞白血病、特定亚型B细胞淋巴瘤和多发性骨髓瘤。然而，CAR-T细胞疗法在应用过程中面临诸多挑战，如在治疗血液系统肿瘤中的抵抗、生产周期长、个体化/价格昂贵，在实体瘤中的肿瘤异质性强/抗原逃逸、浸润能力不足、免疫抑制微环境和反应性差等问题。随着肿瘤免疫学研究的深入和基因工程技术的发展，尝试了众多新策略来提升CAR-T细胞疗法的疗效和普适性。作者根据自身对该领域研究的认知，针对CAR-T细胞疗法的临床关键问题及其应对解决策略进行述评，为未来CAR-T细胞疗法的基础研究和临床转化提供重要思路。

Objective:To investigate the correlation between the lipocalin-2 (LCN-2) level and white matter hyperintensities (WMHs) in patients with ischemic stroke.Methods:Consecutive patients with ischemic stroke admitted to the Department of Neurology, Jinling Hospital, Medical School of Nanjing University from September 2021 to November 2021 and whose duration from onset to hospitalization <14 d were prospectively enrolled. Enzyme-linked immunosorbent assay was used to detect the serum LCN-2. Fazekas scale was used to assess the severity of periventricular and subcortical WMHs. A total WMHs score ≥3 was defined as severe WMHs. Multivariate logistic regression analysis was used to determine the correlation between serum LCN-2 level and WMHs. Results:A total of 179 patients were enrolled, including 122 males (68.2%), aged 64.7±11.6 years. The median serum LCN-2 level was 387.1 g/L, and 86 patients (48.0%) had severe WMHs. Serum LCN-2 in the severe WMH group was significantly higher than that in the non-severe WMH group (505.3±342.4 g/L vs. 367.8±224.5 g/L; t=3.110, P=0.002). Multivariable logistic regression analysis showed that after adjusting for the relevant confounding factors, there was a significant correlation between higher serum LCN-2 and severe WMHs (odds ratio 2.32, 95% confidence interval 1.17-4.63; P=0.017) and higher total WMHs score (odds ratio 1.62, 95% confidence interval 1.12-2.35; P=0.011). Conclusion:Higher serum LCN-2 level is associated with severe WMHs in patients with ischemic stroke.

The biological effects of low-dose radiation (LDR) are still a research hotspot in the field of radiobiology. As research deepens on LDR-induced biological effects and the mechanisms, growing evidence shows that LDR produces distinct biological effects from high-dose radiation, which questions the linear non-threshold model. This article reviews LDR-induced bystander effect, hormesis, adaptive response, and hyper-radiosensitivity, as well as the mechanisms, in order to provide a reference for the transformation of basic research on LDR-related biological effects to clinical application.

N 6-methyladenosine (m 6A) is the most abundant RNA base modification in mammals, especially in eukaryotic messenger RNA (mRNA). N 6-methyladenosine modification can regulate RNA splicing, translocation, stability and ultimately affect protein synthesis. m 6A modification is catalyzed by RNA writers, reduced by erasers and also be recognized by readers. Abnormal changes ofm 6A levels are closely related to tumor occurrence and development, including proliferation, growth, invasion and metastasis. In the process of tumor radiotherapy, m 6A modification affects the efficacy of radiotherapy by affecting DNA damage, tumor stem cell generation and tumor cell radiation sensitivity. This article reviews the role of m 6A-modified epigenetic regulation in malignant tumors and the research progress of its mechanism in tumor radiotherapy, in order to provide new ideas for the development of clinical tumor molecular targeted therapies and radiosensitizers.

Background: and Purpose Paroxysmal atrial fibrillation (PAF) underlying acute stroke frequently evades detection by standard practice, considered to be a combination of routine electrocardiogram (ECG) monitoring, and 24-hour Holter recordings. We hypothesized that nurse-led in-hospital intermittent monitoring approach would increase PAF detection rate. Methods: We recruited patients hospitalised for stroke/transient ischemic attack, without history of atrial fibrillation (AF), in a prospective multi-centre observational study. Patients were monitored using a smartphone-enabled handheld ECG (iECG) during routine nursing observations, and underwent 24-hour Holter monitoring according to local practice. The primary outcome was comparison of AF detection by nurse-led iECG versus Holter monitoring in patients who received both tests: secondary outcome was oral anticoagulant commencement at 3-month following PAF detection. Results: One thousand and seventy-nine patients underwent iECG monitoring: 294 had iECG and Holter monitoring. AF was detected in 25/294 (8.5%) by iECG, and 8/294 (2.8%) by 24-hour Holter recordings (P<0.001). Median duration from stroke onset to AF detection for iECG was 3 days (interquartile range [IQR], 2 to 6) compared with 7 days (IQR, 6 to 10) for Holter recordings (P=0.02). Of 25 patients with AF detected by iECG, 11 were commenced on oral anticoagulant, compared to 5/8 for Holter. AF was detected in 8.8% (69/785 patients) who underwent iECG recordings only (P=0.8 vs. those who had both iECG and 24-hour Holter). Conclusions Nurse-led in-hospital iECG surveillance after stroke is feasible and effective and detects more PAF earlier and more frequently than routine 24-hour Holter recordings. Screening with iECG could be incorporated into routine post-stroke nursing observations to increase diagnosis of PAF, and facilitate institution of guideline-recommended anticoagulation.

Objective To investigate the difference between histopathological changes of brain white matter in low-density lipoprotein receptor (LDLR) homozygous mutation rats with hypercholesterolemia and wild-type rats.Methods Thirty LDLR-/-rats and 28 wild-type rats were selected.Plasma cholesterol levels were measured by enzyme-linked immunosorbent assay at 15,18 and 26 weeks old respectively.The axonal structure of the corpus callosum area was observed by transmission electron microscopy.The myelin basic protein (MBP) of the corpus callosum area was quantitatively analyzed by Western blotting.In addition,at 26 weeks old,the myelin sheaths were stained by fast blue staining.The expression level of MBP in white matter was further detected by immunofluorescence staining,and the morphological changes of glial cells were observed.Results Compared with the wild-type rats,the plasma cholesterol concentration in LDLR-/-rats increased significantly,and it could be as high as 3.3 times at 26 weeks.The results of electron microscopy showed that the LDLR-/-rats had axonal injury at 15 weeks and aggravated gradually over time.At 26 weeks,Western blot analysis of the LDLR-/-rats showed that the MBP expression level of the corpus callosum area decreased significantly.Fast blue staining showed loosening of nerve fibers,diffuse vacuole formation,and myelinated nerve fiber loss in the corpus callosum area.In addition,it was also found that the number of oligodendrocytes in LDLR-/-rats was significantly reduced,and large numbers of astrocytes and microglia were activated.Conclusions LDLR-/-rats will have spontaneous hypercholesterolemia.Axonal injury,demyelination,decreased oligodendrocytes,as well as the abnormal activation of astrocytes and microglia are present in the early adult brain white matter area.

Objective To investigate the effect of human serum albumin (Alb) on the permeability of blood-brain barrier (BBB) after subarachnoid hemorrhage (SAH) and the pathways for Alb uptake in endothelial cells.Methods Mouse brain endothelial cells (bEnd.3) were cultured in the Transwell chamber was used to induce a BBB model.A SAH in vitro model was induced by adding 10 μmol/L oxyhemoglobin into the culture medium.The cells were divided into 3 groups:control group,SAH group,and Alb group (10 mg/ml).Transendothelial electric resistance (TEER) was used to detect the permeability of BBB.A confocal microscope was used to observe whether the fluorescent labeled Alb could be uptaken by bEnd.3cells.Immunoprecipitation was used to detect whether Alb could interact with the cells of caveolin 1 (Cav-1).According to the principle of siRNA,Cav-1 siRNA was transfected into bEnd.3 cells to inhibit the expression of Cav-1.Western blot analysis was used to detect whether bEnd.3 cells could uptake Alb.TEER was used to detect the permeability of BBB.Results Compared with the SAH group,the TEER value of the Alb group increased significantly (P =0.011).Alb was uptaken by bEnd.3 cells and interacted with Cav-1 in bEnd.3 cells.Cav-1 siRNA transfection could significantly inhibit the expression of Cav-1 in bEnd.3cells and reduce the uptake ability of Alb by cells (P=0.025),resulting in a significant decrease in the protective effect of Alb on BBB (P ＜ 0.001).Conclusion Cav-1 may be uptaken by endothelial cells under the participation of Cav-1 and improve the permeability of BBB after SAH.

Objective: To investigate the effect of tumor-associated macrophages on the stemness of esophageal cancer cells and the potential mechanism of antiproliferative effects of aspirin (ASA). Methods: The effects of aspirin on the stemness characteristics of KYSE-450 cells and KYSE-450 cells co-cultured with M2 macrophages (KYSE-450+ M2) were performed using spheroid formation assay. After treatment with aspirin, the expression of different chemokines, the core pluripotency gene Nanog and the stem cell marker CD90 in different cell groups were determined by real-time quantitative PCR, flow cytometry and Western blot. Results: The number of spheres formed in the ASA and KYSE-450+ M2 cell groups were 7.00±1.23 and 34.33±2.33, respectively, showing statistically significant difference compared with that of control group (14.50±2.33, all P<0.05). The number of spheres in KYSE-450+ M2+ ASA cell group were 20.67±2.33, which was significantly lower than that of KYSE-450+ M2 group (P<0.05). The expression levels of Nanog gene in control and ASA groups were 1.00 and 0.50±0.10, respectively, and the difference was statistically significant (P<0.05). Moreover, the expression of Nanog gene in cells of KYSE-450+ M2 group and M2+ KYSE-450+ ASA group was 1.74±0.13 and 1.43±0.05, showing statistically significant difference (P<0.05). When chemokine CCL2 was knocked down, the levels of Nanog gene in M2+ shCCL2-KYSE450+ ASA group and M2+ shCCL2-KYSE450 group were decreased to 1.22±0.11 and 1.17±0.08, respectively, and there was no statistically significant difference between them (P=0.69). Flow cytometry analyses showed that the expression levels of CD90 in control and ASA cells were (2.93±0.52)% and (1.30±0.17)%, respectively, and the difference was statistically significant (P<0.05). Moreover, the expression levels of CD90 in M2+ shCCL2-KYSE450 cells and M2+ shCCL2-KYSE450+ ASA cells were (4.07±0.12)% and (4.73±0.38)%, respectively, showing no statistically significant difference (P=0.17). Conclusions Tumor-associated macrophages enhances the stemness of esophageal cancer cells, whereas aspirin attenuates the stemness by suppressing the expression of CCL2. Aspirin plays an anti-tumor effect in esophageal cancer cells.

White matter lesion is a major subtype of cerebral small vessel disease. Its pathophysiology and mechanism remain unclear. Because the risk factors often coexist in clinical research, it is difficult to judge the relationship between certain risk factors and white matter injury. Moreover, due to the differences in animal and human brain tissue structure, there is currently a lack of reproducible animal models of white matter lesions. Therefore, establishing a practical animal model and further exploring the pathogenesis and risk factors of white matter lesions from the basic research level is crucial for the preclinical study of the treatment of white matter damage. This article reviews the characteristics, optimization measures, and application prospects of the white matter lesion models.

Objective To investigate the relationship between the expression of 06-methyl guanine DNA methyltransferase (MGMT),X-ray repair cross complementation gene 1 (XRCC1) and the incidence of glioma.Methods From February 2015 to September 2017,53 glioma patients (glioma group) in our hospital were enrolled in the study.50 patients with hypertensive intracerebral hemorrhage were selected as control A group,and 106 healthy volunteers as control B group.Immunohistochemical staining was used to detect the expression of MGMT and XRCC1 in brain tissue of glioma group and control A group,and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) were used to detect the polymorphism of MGMT and XRCC1 gene in glioma group and control B group.Results The positive expression rates of MGMT and XRCC1 in the tissues of brain glioma were 47.17％ and 39.62％,respectively,which were significantly higher than those in the control A group (P ＜ 0.05).There was no significant difference in the positive expression rate of MGMT and XRCC1 in patients with grade Ⅰ,,Ⅲ and Ⅳ (P ＞ 0.05);There was no correlation between the expression of MGMT and XRCC1 in glioma tissues (rs =0.162,P ＞ 0.05);The proportion of XRCC1 genotype AG + GG in brain glioma group was 58.49％,which was significantly higher than that of control B group (P ＜ 0.05);The proportion of MGMT genotype LP + PP in brain glioma group was 28.30％,which was significantly higher than that of control B group (P ＜ 0.05).Conclusions MGMT and XRCC1 are increased significantly in glioma brain tissues,but not correlatedwith pathological grades;The polymorphism of MGMT and XRCC1 genes may be related to the susceptibilityof gliomas.