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.

Purpose: The poor retention and ambiguous differentiation of stem cells (SCs) within corpus cavernosum (CC) limit the cell application in erectile dysfunction (ED). Herein, the effects and mechanism of microRNA-145 (miR-145) gene modification on modulating the traits and fate of bone marrow-derived mesenchymal stem cells (BMSCs) were investigated. Materials and Methods: The effects of miR-145 on cell apoptosis, proliferation, migration, and differentiation were determined by flow cytometry, cell counting kit-8, transwell assays and myogenic induction. Then, the age-related ED rats were recruited to four groups including phosphate buffer saline, BMSC, vector-BMSC, overexpressed-miR-145-BMSC groups. After cell transplantation, the CC were harvested and prepared to demonstrate the retention and differentiation of BMSCs by immunofluorescent staining. Then, the target of miR-145 was verified by quantitative real-time polymerase chain reaction and immunohistochemical. After that, APTO-253, as an inducer of Krüppel-like factor 4 (KLF4), was introduced for rescue experiments in corpus cavernosum smooth muscle cells (CCSMCs) under the co-culture system. Results: In vitro, miR-145 inhibited the migration and apoptosis of BMSCs and promoted the differentiation of BMSCs into smooth muscle-like cells with stronger contractility. In vivo, the amount of 5-ethynyl-2′-deoxyuridine (EdU)+cells within CC was significantly enhanced and maintained in the miR-145 gene modified BMSC group. The EdU/CD31 co-staning was detected, however, no co-staining of EdU/α-actin was observed. Furthermore, miR-145, which secreted from the gene modified BMSCs, dampened the expression of KLF4. However, the effects of miR-145 on CCSMCs could be rescued by APTO-253. Conclusions Overall, miR-145 modification prolongs the retention of the transplanted BMSCs within the CC, and this effect might be attributed to the modulation of the miR-145/KLF4 axis. Consequently, our findings offer a promising and innovative strategy to enhance the local stem cell-based treatments.

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.

Objective:To investigate the expression of the B cell ectopic gene 2 (BTG2) in the pancreatic cancer tissue and analyze its relationship with the clinicopathological features and prognosis.Methods:46 pairs of pancreatic cancer tissues and corresponding adjacent tissues kept in paraffin in the pathology department, and 9 fresh pancreatic cancer tissues and corresponding adjacent tissues resected by surgery in Department of Pancreatic Surgery of Sinopharm Dongfeng General Hospital from June 2015 to December 2020 were collected. BTG2 gene expression in 46 pairs of pancreatic cancer tissues and corresponding adjacent tissues were detected by immunohistochemical staining, and high and low BTG2 expression groups were divided. BTG2 gene expression in 9 fresh pancreatic cancer tissues and corresponding adjacent tissues were detected by RT-PCR. The correlation between BTG2 protein expression level and clinicopathological features was analyzed. Furthermore, the survival curve and death risk curve were drawn using the Kaplan-Meier method, and the Cox regression hazards model was applied for the univariate and multivariate analysis of the factors affecting the prognosis of pancreatic cancer.Results:29 of 46 (63.04%) pancreatic cancer tissues had high BTG2 expression, and 38(82.61%) of corresponding adjacent tissues had high BTG2 expression; and BTG2 high expression rate of adjacent tissues was significantly higher than that of cancer tissues. Three out of 9 pancreatic cancer tissues were highly differentiated, and six cases had medium-and low differentiation. The BTG2 expression of highly differentiated pancreatic carcinoma was significantly higher than that of moderately and poorly differentiated carcinoma tissues [(0.66±0.07 vs 0.24±0.18); the expression level of adjacent tissues was significantly higher than that of cancer tissues (1.00±0.00 vs 0.38±0.30), and all differences were statistically significant (all P values <0.001). Low BTG2 expression in pancreatic cancer was associated with low tumor differentiation and vascular invasion (all P values <0.05), but was not correlated with tumor location, volume, lymph node metastasis, CA19-9 level and postoperative liver metastasis. The median survival of high BTG2 expression group was significantly longer than that of low BTG2 expression group (525 d vs 266 d, P<0.001). Among patients with survival time ≥300 d, the survival time was significantly higher in the high BTG2 expression group than in the BTG2 low expression group (616±135d vs 426±113 d), and the difference was statistically significant ( P<0.001). Among patients with survival time <300 d, there was no significant difference between BTG2 high and low expression group. The results of the univariate analysis showed that tumor differentiation degree, vascular invasion, BTG2 expression, CA19-9 levels, and postoperative liver metastasis were all associated with the prognosis of pancreatic cancer. The results of the multivariate analysis showed that BTG2 expression level ( HR=2.572, 95% CI1.140-5.802, P=0.023), vascular invasion ( HR=0.023, 95% CI0.072-0.572, P=0.003) and postoperative liver metastasis ( HR=0.240, 95% CI0.102-0.564, P<0.001) were independent risk factors affecting the prognosis of patients with pancreatic cancer. Conclusions:BTG2 expression in pancreatic cancer tissues was significantly lower than that in adjacent tissues, and its low expression was associated with strong aggressiveness, low differentiation degree and poor prognosis of pancreatic cancer. The effect of BTG2 on the prognosis in pancreatic cancer patients was mainly in the long term.

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.

[This corrects the article DOI: 10.1016/j.apsb.2019.03.006.].

Purpose: This study aimed to investigate the clinicopathologic features and mutational landscape of patients with hepatitis B virus (HBV)–related advanced hepatocellular carcinomas (HCC) undergoing transcatheter arterial chemoembolization (TACE). Materials and Methods: From January 2017 to December 2018, 38 patients newly diagnosed with HBV-related advanced HCC were enrolled in the final analysis. Their pathological tissues and corresponding blood samples before TACE treatment were collected for whole-exome sequencing. Response to TACE was evaluated at 1-3 months after two consecutive use of TACE. Predictive factors were analyzed by univariate and multivariate analyses in a bivariate Logistic regression model. Enrichment of related pathways of all driver genes were acquired using the gene set enrichment analysis (GSEA). Results: Among 38 patients, 23 (60.5%) exhibited TACE failure/refractoriness. Patients with TACE failure/refractoriness showed higher frequency of TP53 mutation than their counterparts (p=0.020). Univariate and multivariate analyses showed that only vascular invasion and TP53 mutation were significantly correlated with TACE failure/refractoriness in HBV-related advanced HCC. Of the 16 patients without vascular invasion, eight (50.0%) had TP53 mutations, and TP53 mutation was associated with TACE failure/refractoriness (p=0.041). Moreover, GSEA showed that mitogen-activated protein kinase and apoptosis pathways induced by TP53 mutation were possibly associated with TACE failure/refractoriness. Conclusion Our study suggested that TP53 mutation was independently related with TACE efficacy, which may work via mitogen-activated protein kinase and apoptosis pathways. These findings may provide evidence to help distinguish patients who will particularly benefit from TACE from those who require more personalized therapeutic regimens and rigorous surveillance in HBV-related advanced HCC.

Objective Toevaluatehysterosalpingography(HSG)and MRImanifestationsofintrauterineadhesion(IUA).Methods 40patientswithIUAconfirmedbyhysteroscopyunderwentHSGand MRIexamination.Wefoundoutthefeaturesofthoseimages, andworkedoutthedetectablerateofeachexamination.Results Inall40patients,19patientsweremildIUAprovedbyhysteroscopy,in which10patientswerediscoveredbyHSG,10patientsby MRIT2WI,16patientsby MRICUBE.16patientsweremiddleIUA,in which9patientswerediscoveredbyHSG,10patientsbyMRIT2WI,16patientsbyMRICUBE.5patientsweresevereIUA,allpatients werediscoveredbyHSG,MRIT2WIand MRICUBE.Andinallcases,therewere12casesofmembraneadhesions,inwhich5cases wereshowninHSG,8casesinMRIT2WIand10casesinMRICUBE.23caseswerefibrousadhesions,inwhich12caseswereshown in HSG,17casesinMRIT2WIand22casesinMRICUBE.5patientsweremuscularadhesions,allpatientswerediscoveredbyHSG, MRIT2WIandMRICUBE.Conclusion ComparingtoHSG,MRIexaminationshowshigherdetectablerateofIUA.MRICUBEsequences playsanimportantpartinthediagnosisofIUA.

Objective To explore the CT dominant phase and optimal classification model in differenting clear cell renal cell carcinoma (ccRCC) from fat‐poor angiomyolipoma (fpAML) through quantitative multiple‐phase CT radiomic features analysis. Methods Clinical and imaging data of 195 cases pathologically confirmed ccRCC (n=131) and fpAML (n=64) were retrospectively studied. All the patients underwent non‐contrast enhanced CT scans and dynamic multi‐phase (corticomedullary phase, medullary phase and excretion phase) contrast‐enhanced CT scans. Regions of interest (ROIs) were manually delineated based on the selected image slices with the maximal diameter of the lesion using ITK‐SNAP software, followed by the acquisition of candidate CT radiomic feature sets from each phase with statistically significant differences by using Mann‐Whitney U test. Then, using the synthetic minority oversampling technique (SMOTE), 232 classification models which are composed of 29 different feature selection algorithms (top 10 features were chosen by the backward elimination method) and 8 different classifiers were constructed. Employing the 5‐fold cross‐validation method, the performance of each classification models for each phase was evaluated using accuracy (ACC), sensitivity (SEN), specificity (SPE) and area under receiver operating characteristic curve (AUC), to acquire dominant CT phases and the optimal classification models for distingushing ccRCC and fpAML, along with the key imaging radiomic features. Results In this study, the mean maximal diameter of ccRCC and fpAML lesions were (3.9±1.4) cm, and (3.5±1.7) cm, respectively, and there was no statistically significant difference in the size of the tumor between two groups (P>0.05). From 102 initial imaging feature sets, the total number of candidate imaging feature sets (P<0.05) were:non‐enhanced phase (n=26), corticomedullary phase (n=71), medullary phase (n=68), excretion phase (n=62). Among the 232 classification models through different combination of classifiers and feature selectors, the amount of classification models which achieved the maximum of AUC value (AUCmax) from different CT phases were: non‐enhanced phase (n=106, 45.7%), corticomedullary phase (n=94, 40.5%), medullary phase (n=23, 9.9%), excretion phase (n=9, 3.9%). Imaging features from non‐enhanced phase and corticomedullary phase yielded higher performance compared with medullary phase and excretion phase, with the corresponding optimal prediction models were SVM‐fisher_score (AUC: 0.897, ACC: 83%, SEN: 84%, SPE:80%) and Logistic Regression‐RFS (AUC: 0.891, ACC: 83%, SEN: 81%, SPE: 89%), respectively. Conclusions The quantitative imaging features from non‐enhanced and corticomedullary phase have better performance among proposed classification models than that from medullary phase and excretion phase. Furthermore, it is feasible to acquire proper combination of feature selection and classifiers to achieve high performance in identifying ccRCC and fpAML.