Chimeric antigen receptor T (CAR-T) lymphocytes are at the forefront of adoptive immunotherapy research, and this technology has significantly advanced the prospects of tumor immunotherapy. CAR-T therapy has demonstrated remarkable efficacy in haematological tumours of lymphoid origin and provided therapeutic possibility for solid tumours. Currently, CAR-T cell preparation predominantly involves transfection of T cells with viral vectors. However, the production of viral vectors is time-consuming, expensive, and the vectors have low loading capacity, along with insertion instability. Consequently, there is a pressing need to develop more convenient and precise non-viral gene delivery methods. This paper reviews the most promising non-viral gene delivery technologies, including CRISPR/Cas9 gene editing, transposon systems such as Sleeping Beauty (SB) and PiggyBac (PB), and mRNA, and anticipates the future development of non-viral vector-based CAR-T therapies.
Humans ; Immunotherapy, Adoptive/methods* ; Receptors, Chimeric Antigen/immunology* ; Animals ; Gene Transfer Techniques ; Genetic Vectors/genetics* ; Gene Editing ; CRISPR-Cas Systems/genetics* ; DNA Transposable Elements/genetics* ; T-Lymphocytes/immunology* ; Neoplasms/immunology*

Immunotherapy has emerged as a revolutionary approach to treat immune-related diseases. Dendritic cells (DCs) play a pivotal role in orchestrating immune responses, making them an attractive target for immunotherapeutic interventions. Modulation of gene expression in DCs using genome editing techniques, such as the CRISPR-Cas system, is important for regulating DC functions. However, the precise delivery of CRISPR-based therapies to DCs has posed a significant challenge. While lipid nanoparticles (LNPs) have been extensively studied for gene editing in tumor cells, their potential application in DCs has remained relatively unexplored. This study investigates the important role of cholesterol in regulating the efficiency of BAMEA-O16B lipid-assisted nanoparticles (BLANs) as carriers of CRISPR/Cas9 for gene editing in DCs. Remarkably, BLANs with low cholesterol density exhibit exceptional mRNA uptake, improved endosomal escape, and efficient single-guide RNA release capabilities. Administration of BLAN_mCas9/gPD-L1 results in substantial PD-L1 gene knockout in conventional dendritic cells (cDCs), accompanied by heightened cDC1 activation, T cell stimulation, and significant suppression of tumor growth. The study underscores the pivotal role of cholesterol density within LNPs, revealing potent influence on gene editing efficacy within DCs. This strategy holds immense promise for the field of cancer immunotherapy, offering a novel avenue for treating immune-related diseases.

Objective To understand and assess the quality status of national drug sampling inspection in 2022 and the possible risks of drug quality.Methods With the aim of identifying issues and preventing risks,a sampling model of"dispersed sampling,centralized inspection,exploratory research,and comprehensive evaluation"was used to review the historical inspection data of national drug sampling.The overall quality and potential risks of national drug sampling in 2022 were analyzed,with a focus on identifying and examining the main quality problems and risks.Results The average pass rate of the sampling inspection in 2022 was 99.4% .The overall safety situation of China's post-marketing drug quality is stable and manageable,indicating that the drug quality is at a high level.Conclusions By carrying out exploratory studies related to drug safety,authenticity,and efficacy,national sampling plays an important role in comprehensively evaluating the quality status of drugs.It aids in improving the quality control level of drug varieties and industry standards,strengthening quality and safety supervision,combating illegal practices such as adulteration and falsification,and providing public warnings about drug safety.

Cryoablation (CRA) and microwave ablation (MWA) are two main local treatments for hepatocellular carcinoma (HCC). However, which one is more curative and suitable for combining with immunotherapy is still controversial. Herein, CRA induced higher tumoral PD-L1 expression and more T cells infiltration, but less PD-L1^highCD11b⁺ myeloid cells infiltration than MWA in HCC. Furthermore, CRA had better curative effect than MWA for anti-PD-L1 combination therapy in mouse models. Mechanistically, anti-PD-L1 antibody facilitated infiltration of CD8⁺ T cells by enhancing the secretion of CXCL9 from cDC1 cells after CRA therapy. On the other hand, anti-PD-L1 antibody promoted the infiltration of NK cells to eliminate PD-L1^highCD11b⁺ myeloid cells by antibody-dependent cell-mediated cytotoxicity (ADCC) effect after CRA therapy. Both aspects relieved the immunosuppressive microenvironment after CRA therapy. Notably, the wild-type PD-L1 Avelumab (Bavencio), compared to the mutant PD-L1 atezolizumab (Tecentriq), was better at inducing the ADCC effect to target PD-L1^highCD11b⁺ myeloid cells. Collectively, our study uncovered the novel insights that CRA showed superior curative effect than MWA in combining with anti-PD-L1 antibody by strengthening CTL/NK cell immune responses, which provided a strong rationale for combining CRA and PD-L1 blockade in the clinical treatment for HCC.

Abstract: Objective　To investigate the clinical manifestations, diagnosis, treatment and prognosis of COVID-19-associated multisystem inflammatory syndrome in children (MIS-C) with multisystem organ failure, and to analyze the differences between the syndrome resembling Kawasaki disease and Kawasaki disease. Methods We report a case of child diagnosed with SARS-CoV-2 infection who developed MIS-C with multi-organ failure. Literature was reviewed to further discuss clinical symptoms, diagnosis, treatment, and prognosis of MIS-C. Results The patient 's condition progressed rapidly and was severe, with acute kidney injury at admission, presenting with urine output of 0 mL in 8 hours, serum creatinine of 675.1 μmol/L, urea nitrogen of 38.2 mmol/L, creatine kinase 4 932 U/L, along with continuously elevated serum amylase, peaking at 5 809.7 U/L, aspartate aminotransferase of 995 U/L, alanine aminotransferase of 240 U/L, and frequent vomiting. The patient was categorized as critically ill. The confirmed patient with MIS-C was managed in pediatric intensive care units (PICU), received early continuous blood purification, jejunal tube feeding, and combined treatment with intravenous methylprednisolone pulse and human immunoglobulin for intravenous injection (IVIG), resulting in complete recovery and discharge. Through literature review, it can be analyzed that MIS-C is systemic inflammatory response characterized by multi-organ dysfunction from a cytokine storm, which differentiates it from Kawasaki disease characterized by acute systemic vasculitis triggered by infectious factors, ultimately leading to cardiovascular accidents as the main cause of death.Conclusions When fever (≥38 ℃) persists for at least 24 hours, accompanied by high inflammatory symptoms and dysfunction in two or more systems, requiring intensive care treatment, and there is evidence of SARS-CoV-2 infection, MIS-C should be diagnosed early, and intervention and treatment should be initiated promptly to achieve favorable outcomes.

OBJECTIVE To put forward suggestions on strengthening the quality supervision of Chinese patent drugs and preventing and controlling safety risks by analyzing the current quality situation and quality risks of sampling inspection of Chinese patent drugs. METHODS Reviewed the quality status of Chinese patent drugs for national drug sampling and testing over the past years, introduced the supervision concept and mode of Chinese patent drugs, conducted an overall analysis of the overall quality of Chinese patent drugs for national drug sampling and testing in 2021, focused on the mining and analysis of the main quality problems and risks identified. RESULTS Although the qualified rate of sampling inspection of Chinese patent medicines had shown a positive trend year by year, through exploratory research, it was found that there were still some potential quality risks in the quality of Chinese patent medicines in terms of process prescriptions, internal control standards, raw medicinal materials, excipients, harmful substance residues, illegal addition, etc. CONCLUSION Drug quality sampling and testing plays an important role in strengthening the quality and safety supervision of traditional Chinese medicine. According to the characteristics of different industrial chains of traditional Chinese medicine, different regulatory strategies should adopt, build a more scientific and reasonable regulatory model for Chinese patent medicine, and improve the quality control level and industry standards of Chinese patent medicine.

Objective:To evaluate the value of enhanced CT radiomics feature model for predicting 5-year overall survival (OS) of esophageal squamous cell carcinoma patients after radiotherapy.Methods:Clinical data of 218 patients with esophageal squamous cell carcinoma treated with radical chemoradiotherapy in the Fourth Hospital of Hebei Medical University from July 2016 to December 2017 were retrospectively analyzed. Patients were randomly divided into the training group ( n=153) or a validation group ( n=65) at a 7 vs. 3 ratio. Enhanced CT radiomics features were extracted. The data in the training group was used to construct the prediction model, and the data in the validation group were utilized to validate the efficiency of this model for predicting the 5-year OS of patients. The predictive performance of this model was assessed by the receiver operating characteristic (ROC) curve, consistency index (C-index), and decision curve analysis (DCA). Results:The 1-, 3-, 5-year OS rates were 67.0%, 33.4%, 24.9%. Five radiomic features were selected from extracted features in the training group to construct the radiomic signature (RS) for predicting 5-year OS. The area under the ROC curve (AUC) was 0.760 in the training group and 0.707 in the validation group, and the C-index was 0.680 and 0.684, respectively. The radiomics nomogram, which incorporated the RS with clinical risk factors, were established to predict the 5-year OS of esophageal squamous cell carcinoma patients after radiotherapy. The AUC was 0.782 in the training group and 0.751 in the validation group, and the C-index was 0.708 and 0.688, respectively. According to the optimal cutoff of the model, all patients were divided into the high risk and low risk groups. The 1-, 3-, 5-year OS rates were 86.5%, 65.4%, 28.9% in the low risk group, and 58.4%, 17.8%, 5.9% in the high risk group, and the differences were statistically significant (all P<0.001). Similar conclusions were obtained in the validation group (all P<0.001). Conclusion:Enhanced CT radiomics features can be utilized to construct the prediction model for 5-year OS of esophageal squamous cell carcinoma patients after radiotherapy, which can be applied in clinical practice.

Objective:To investigate the effect of miRNA-296-5p (miR-296-5p) on the migration and invasion of hypoxia-induced pancreatic cancer cells and its related mechanisms.Methods:Human pancreatic cancer cell line PANC-1 was selected. Pancreatic cancer tissues from 55 pancreatic cancer patients who underwent the resection and adjacent carcinoma normal pancreatic tissues from 10 patients at Shanghai Fengxian District Central Hospital and Bengbu Medical College First Affiliated Hospital between January 2010 and December 2014 were collected. The expression levels of hypoxia-inducible factor 1α (HIF-1α) and miR-296-5p in tissue microarray of pancreatic cancer and adjacent carcinoma normal pancreatic tissues were detected by using immunohistochemistry and in situ hybridization. The relationship between miR-296-5p and HIF-1α as well as their correlation with clinicopathological characteristics of patients were analyzed. PANC-1 cells were divided into hypoxic group and normoxic group. Transwell assay was used to detect the cell migration and invasion ability of both groups. Real-time quantitative polymerase chain reaction (qRT-PCR) was used to examine the expressions of HIF-1α and miR-296-5p under hypoxic environment of both groups. The expression of HIF-1α was interfered by transfecting small interfering RNA (siRNA). PANC-1 cells were divided into PANC-1 group (the empty control), PANC-1-NC group (the negative control) and PANC-1-siRNA group. The expression of miR-296-5p was measured. After co-transfecting miR-296-5p agonist and miR-296-5p inhibitor, the cells were divided into Agomir-miR-296-5p group (agonist group), Agomir-miR-296-5p-NC group (agonist negative control group), Antagomir-miR-296-5p group (inhibitor group) and Antagomir-miR-296-5p-NC group (inhibitor negative control group). Transwell assay was used to detect the cell migration and invasion ability of all groups. Luciferase reporter gene system was used to verify whether miR-296-5p promoter region had binding site of HIF-1α.Results:The high expression rate of HIF-1α in pancreatic cancer tissues was higher than that of adjacent carcinoma normal pancreatic tissues [81.8% (45/55) vs. 0 (0/10), P<0.01], and the high expression rate of miR-296-5p in pancreatic cancer tissues was lower than that of adjacent carcinoma normal pancreatic tissues [12.7% (7/55) vs. 90.0% (9/10), χ2 = 27.23, P<0.01]. The expression of HIF-1α was negatively correlated with that of miR-296-5p ( r = -0.53, P<0.01). The low expression of miR-296-5p was closely related with the tumor diameter, TNM staging, lymph node metastasis (all P<0.05). The number of PANC-1 invasion cell was 15.3±2.1 in normoxic group and 24.7±1.5 in hypoxic group, and the difference was statistically significant ( t = 0.26, P = 0.003). The number of PANC-1 migration cell was 20.7±3.8 in hypoxic group and 32.7±1.2 in normoxic group, and the difference was statistically significant ( t = 5.25, P = 0.006). The relative expression level of HIF-1α mRNA in PANC-1 cell of hypoxic group was higher than that of normoxic group [(1.00±0.01) vs. (0.30±0.02)], and the difference was statistically significant ( t = 56.45, P<0.01); the relative expression level of miR-296-5p in PANC-1 cell of hypoxic group was lower than that of normoxic group [(1.14±0.04) vs. (3.05±0.20)], and the difference was statistically significant ( t = 16.05, P<0.01). The number of invasion cells in PANC-1 group, PANC-1-NC group and PANC-1-siRNA group was 24.7±1.5, 25.7±1.5, 12.0±1.7, respectively, and the difference was statistically significant ( F = 68.13, P<0.01).The cell invasion ability in PANC-1-siRNA group was decreased compared with that in PANC-1 group ( t = 9.50, P = 0.001). The number of cell migration was 32.7±1.2, 37±1.0, 17.3±1.2, respectively in PANC-1 group, PANC-1-NC group and PANC-1-siRNA group, and the difference was statistically significant ( F = 262.09, P<0.01). The cell migration ability in PANC-1-siRNA group was decreased compared with that in PANC-1 group ( t = 16.26, P<0.01). The cell invasion and migration ability in Antagomir-miR-296-5p group was increased compared with that in PANC-1 group (all P<0.05); the cell invasion and migration ability in Agomir-miR-296-5p group was decreased compared with that in PANC-1 group (all P<0.05). The results of luciferase activity detected by luciferase reporter gene system showed that miR-296-5p had the target binding to HIF-1α. Conclusions:HIF-1α plays a key role in the invasion and migration of hypoxia-induced pancreatic cancer cells through negatively reducing miR-296-5p.

Objective:To summarize the perinatal management and prognosis of congenital chylous ascites (CCA).Methods:Clinical data of 20 infants diagnosed with CCA and treated in Guangdong Women and Children Hospital from June 2015 to November 2020 were retrospectively analyzed and described.Results:There were ten patients with isolated CCA and ten with non-isolated CCA. In isolated CCA cases, seven were cured after conservative treatment and three after postoperative conservative treatment following an ineffective surgery. Non-isolated CCA cases were complicated by intrauterine cytomegalovirus infection ( n=1), malrotation of intestine ( n=4) or bilateral chylothorax ( n=5). In addition to conservative treatment for CCA, non-isolated CCA patients also received antiviral therapy, Ladd's procedure or intrauterine/extrauterine closed thoracic drainage. Of eight patients who were firstly diagnosed with ascites before 30 gestational weeks, including four isolated and four non-isolated cases, only one underwent surgical intervention. During hospitalization, serious infections occurred in three infants with isolated CCA and two with non-isolated CCA, and were all controlled by anti-infection treatment. During a follow-up to median age of 29 months (15-82 months), none of the patients had any abnormalities except for the one with intrauterine cytomegalovirus infection who was deaf at the age of two. Conclusions:Conservative management is effective and the prognosis is generally good for most cases with isolated CCA. Treatment and prognosis of non-isolated CCA depend on its comorbidities. Gestational age at diagnosis may not be a prognostic predictor.

Background::Joint dislocations significantly impact public health. However, a comprehensive study on the incidence, distribution, and risk factors for joint dislocations in China is lacking. We conducted the China National Joint Dislocation Study, which is a part of the China National Fracture Study conducted to obtain the national incidence and risk factors for traumatic fractures, and to investigate the incidence and risk factors for joint dislocations.Methods::For this national retrospective epidemiological study, 512,187 participants were recruited using stratified random sampling and probability-proportional-to-size method from January 19 to May 16, 2015. Participants who sustained joint dislocations of the trunk, arms, or legs (skull, sternum, and ribs being excluded) in 2014 were personally interviewed to obtain data on age, educational background, ethnic origin, occupation, geographic region, and urbanization degree. The joint-dislocation incidence was calculated based on age, sex, body site, and demographic factors. The risk factors for different groups were examined using multiple logistic regression.Results::One hundred and nineteen participants sustained 121 joint dislocations in 2014. The population-weighted incidence rate of joint dislocations of the trunk, arms, or legs was 0.22 (95% confidence interval [CI]: 0.16, 0.27) per 1000 population in 2014 (men, 0.27 [0.20, 0.34]; women, 0.16 [0.10, 0.23]). For all ages, previous dislocation history (male: OR 42.33, 95% confidence interval [CI]: 12.03–148.90; female: OR 54.43, 95% CI: 17.37–170.50) and alcohol consumption (male: OR 3.50, 95% CI: 1.49–8.22; female: OR 2.65, 95% CI: 1.08–6.50) were risk factors for joint dislocation. Sleeping less than 7 h/day was a risk factor for men. Compared with children, women aged ≥15 years (female 15–64 years: OR 0.16, 95% CI: 0.04–0.61; female ≥65 years: OR 0.06, 95% CI: 0.01–0.41) were less likely to sustain joint dislocations. Women with more than three children were at higher dislocation risk than women without children (OR 6.92, 95% CI: 1.18–40.78).Conclusions::The up-to-date data on joint dislocation incidence, distribution, and risk factors can be used as a reference for national healthcare, prevention, and management in China. Specific strategies for decreasing alcohol consumption and encouraging adequate sleeping hours should be developed to prevent or reduce dislocation incidents.Trial Registration::Chinese Clinical Trial Registry, ChiCTR-EPR-15005878.