OBJECTIVES: This study investigated the effects of a polycaprolactone (PCL)-polyethylene glycol (PEG) scaffold incorporated with concentrated growth factor (CGF) on the adhesion, proliferation, and osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs). METHODS: The PCL-PEG-CGF composite scaffold was fabricated using an immersion and freeze-drying technique. Its microstructure, mechanical properties, and biocompatibility were systematically characterized. The hPDLSCs were isolated through enzymatic digestion, and the hPDLSCs were identified through flow cytometry. Third-passage hPDLSCs were seeded onto the composite scaffolds, and their adhesion, proliferation and osteogenic differentiation were assessed using CCK-8 assays, 4',6-diamidino-2-phenylindole (DAPI) staining, alkaline phosphatase (ALP) staining, alizarin red staining, and Western blot analysis of osteogenesis-related proteins [Runt-related transcription factor 2 (Runx2), ALP, and morphogenetic protein 2 (BMP2)]. RESULTS: Scanning electron microscopy revealed that the PCL-PEG-CGF composite scaffold exhibited a honeycomb-like structure with heterogeneous pore sizes. The composite scaffold exhibited excellent hydrophilicity, as evidenced by a contact angle (θ) approaching 0° within 6 s. Its elastic modulus was measured at (4.590 0±0.149 3) MPa, with comparable hydrophilicity, fracture tensile strength, and fracture elongation to PCL-PEG scaffold. The hPDLSCs exhibited significantly improved adhesion to the PCL-PEG-CGF composite scaffold compared with the PCL-PEG scaffold (P<0.01). Additionally, cell proliferation was markedly improved in all the experimental groups on days 3, 5, and 7 (P<0.01), and statistically significant differences were found between the PCL-PEG-CGF group and other groups (P<0.01). The PCL-PEG-CGF group showed significantly elevated ALP activity (P<0.05), increased mineralization nodule formation, and upregulated expression of osteogenic-related proteins (Runx2, BMP2 and ALP; P<0.05). CONCLUSIONS The PCL-PEG-CGF composite scaffold exhibited excellent mechanical properties and biocompatibility, enhancing the adhesion and proliferation of hPDLSCs and promoting their osteogenic differentiation by upregulating osteogenic-related proteins.
Humans ; Polyesters/chemistry* ; Periodontal Ligament/cytology* ; Polyethylene Glycols/chemistry* ; Stem Cells/cytology* ; Tissue Scaffolds ; Cell Proliferation ; Osteogenesis ; Cell Differentiation ; Cell Adhesion ; Bone Morphogenetic Protein 2/metabolism* ; Cells, Cultured ; Alkaline Phosphatase/metabolism* ; Core Binding Factor Alpha 1 Subunit/metabolism* ; Intercellular Signaling Peptides and Proteins/pharmacology* ; Tissue Engineering/methods*

Objective:To investigate the etiology, clinical features and outcome of isolated anterior cerebral artery (ACA) territory infarction.Methods:Patients with isolated ACA territory infarction admitted to the Affiliated Shuyang Hospital of Xuzhou Medical University from April 2019 to December 2023 were retrospectively included. The demographic and clinical data were collected. The etiology subtypes of stroke were divided into large artery atherosclerosis (LAA), cardioembolism (CE), small vessel occlusion (SVO), stroke of other determined etiology (SOE), and stroke of undetermined etiology (SUE). At 90 days after onset, the modified Rankin Scale was used to evaluate the outcome. 0~2 points were defined as good outcome, and >2 points were defined as poor outcome. Multivariate logistic regression analysis was used to identify independent risk factors for poor outcomes. Results:A total of 67 patients with isolated ACA territory infarction were enrolled, including 31 males (46.27%), aged 67.20±12.59 years. There were 26 patients (38.81%) of LAA type, 10 (14.93%) of CE type, 7 (10.45%) of SVO type, 12 (17.91%) of SOE type, and 12 (17.91%) of SUE type. Fifty-two patients (77.61%) had good outcome, while 15 (22.39%) had poor outcome. There were significant differences in gender, diabetes, atrial fibrillation, antithrombin-Ⅲ activity, urinary incontinence, cognitive impairment, and the baseline National Institutes of Health Stroke Scale (NIHSS) score among different etiological subtypes (all P<0.05). Multivariate logistic regression analysis showed that previous stroke history (odds ratio 17.995, 95% confidence interval 1.276-253.852; P=0.032) and high baseline NIHSS score (odds ratio 2.094, 95% confidence interval 1.333-3.292; P=0.001) were the independent risk factors for poor outcome. Conclusions:The most common etiology of isolated ACA territory infarction is LAA, and most patients have good outcome. Previous stroke history and high baseline NIHSS score are the independent risk factors for poor outcome.

Objective:To construct a three-dimensional calcaneal finite element model and study its biomechanical distribution by finite element analysis.Methods:Four large calcaneal specimens were taken from the Department of Human Anatomy of Kangda College of Nanjing Medical University in the second semester of 2021—2022. Micro CT scans were performed to obtain the image data of these specimens, and then the three-dimensional calcaneal model was reconstructed using the Mimics and Geomagic Wrap software. The obtained data were imported into the finite element analysis software to perform the material assignments and mesh delineation to obtain the three-dimensional finite element model. The boundary constraints were set, and loads were applied to the calcaneal model to perform finite element analysis calculations, and the stresses and displacements of the finite element model were extracted.Results:The three-dimensional calcaneal model had a high simulation quality. The morphology and structure were complete and without defects, and the size was consistent with that of the real calcaneus, which restores the original bony morphology and structure. The geometric appearance of the three-dimensional calcaneal model after meshing was vivid and lifelike, and without defect. The reconstructed morphology and structure of the three-dimensional model were the same as those of the three-dimensional reconstruction, the size was the same, and the main structure was clearly visible. The meshes and nodes of each component of the three-dimensional calcaneal model were uniformly distributed, and their numbers were comparable to those of the real heel bone. The maximum stresses of the three-dimensional calcaneal model were in the cortical bone, and the maximum stresses were in the same position. These two positions were the lowest part of the anterior part of heel bones. The maximum displacements in the three-dimensional calcaneal model were all less than 0.10 mm.Conclusions:A high-precision three-dimensional finite element calcaneal model has been constructed, which is of good mechanical validity.

Objective:To construct a finite element analysis model for the application of sustentaculum tail sustentacular screw in the treatment of calcaneal fracture of Sanders type Ⅱ. The finite element analysis method was used to study the effectiveness of the simulation treatment of Sanders type Ⅱ calcaneal fracture.Methods:A complete adult calcaneal specimen that was not affecting the normal quality of bone after teaching in the Department of Human Anatomy, Kangda College of Nanjing Medical University from 2021 to 2022 second semester was selected. A micro CT scan was performed to obtain data. The three-dimensional model of the calcaneus was reconstructed by the software of Mimics and Geomagic Wrap. The internal fixation and fracutre model were drawn using SolidWorks software. The obtained data were imported into the finite element analysis software for material assignment and meshing to obtain a three-dimensional finite element model. The load and boundary constraints were applied to each model to perform finite element analysis and calculation, and the stress and displacement of the model were extracted. Finally, the experimental screw placement of the calcaneus loading process was verified.Results:The three-dimensional model of calcaneal fracture had a high degree of appearance simulation, complete morphological structure, no defect, and was consistent with the size of the solid calcaneus. After surgical treatment, the anatomical reduction of each fracture model was good, the steel plate fit to the bone surface, and the screw did not protrude outside the model. There was no deformation or damage to the whole internal fixation, the grid division was clear, and the number was relatively uniform and consistent. The maximum stress in each type of calcaneal fracture model was in the internal fixation, and the overall stress was uniformly distributed in each component. The maximum displacement distance of each type of calcaneal fracture was less than 0.13 mm, and the angles and lengths of nails were accurate. The tip of the Kirschner wires was located in the sustentaculum tail. There was no rupture of the sustentaculum tali and no protrusion of the Kirschner wires.Conclusions:The internal fixation is an effective method to treat the Sanders type Ⅱ calcaneal fractures by using sustentaculum tail sustentacular screws.

The treatment of mediastinal tumor has always been a clinical difficulty due to its complex anatomical location and many important organs.Compared with traditional local treatment,endobronchial ultrasound guided laser multi-point ablation of mediastinal tumors has many advantages,including real-time monitoring of ablation range and effect,avoidance of damage to normal tissue and organs,few side effects and good tolerance.This article describes the standard operating procedure for endobronchial ultrasound guided laser multi-point ablation of mediastinal tumors.

Objective:To investigate the effects of multidisciplinary small-class teaching on expertise and skill acquisition and learning experience in standardized residency training in general surgery.Methods:Sixty residents of grade 2021 rotating in general surgery from January to August 2023 were divided into multidisciplinary teaching group ( n=30) and traditional teaching group ( n=30, to receive tradition one-on-one teaching). All the residents underwent a theoretical examination and Mini-Clinical Evaluation Exercise (Mini-CEX) skill assessment before admission, and the scores were compared between the two groups. At the end of training, the two groups were compared in terms of theoretical and Mini-CEX skill assessment scores and the degree of satisfaction with teaching. Statistical analysis was conducted using SPSS 26.0. Results:There were no significant differences in the theoretical assessment and Mini-CEX skill assessment scores before admission between the two groups ( P>0.05). At the end of training, the multidisciplinary teaching group had a significantly higher theoretical examination score [(88.15±3.45) vs. (72.25±4.36), P<0.05] and a significantly higher Mini-CEX score [(86.35±2.24) vs. (76.28±3.92), P<0.05] compared with the traditional teaching group. According to the survey, the residents in the multidisciplinary teaching group were more satisfied with teaching and more likely to recognize the teaching effects. Conclusions:Multidisciplinary small-class teaching can help improve the quality of standardized general surgery residency training on gastrointestinal tumor treatment, which is a highly accepted and effective attempt at standardized residency training.

Pathological puncture biopsy is the key to early diagnosis and treatment of thoracic tumours,and percutaneous puncture biopsy(PTNB)technology has been widely used in clinical practice.Common complications of PTNB include pneumothorax,air embolism and pulmonary hemorrhage.In order to optimize traditional PTNB,our center has developed a new percutaneous puncture diagnosis and ablation technology for lung lesions,which greatly reduces the occurrence of complications,increases the accuracy of pathological biopsy.At the same time,it can bridge target lesion ablation.This article describes the standard operating procedure for diagnosis and ablation techniques of filling operation assisted percutaneous transthoracic needle biopsy under rigid endoscopy with regular frequency controlled ventilation during general anesthesia.

Recent clinical studies have shown that mutation of phosphatase and tensin homolog deleted on chromosome 10 (PTEN) gene in cancer cells may be associated with immunosuppressive tumor microenvironment (TME) and poor response to immune checkpoint blockade (ICB) therapy. Therefore, efficiently restoring PTEN gene expression in cancer cells is critical to improving the responding rate to ICB therapy. Here, we screened an adeno-associated virus (AAV) capsid for efficient PTEN gene delivery into B16F10 tumor cells. We demonstrated that intratumorally injected AAV6-PTEN successfully restored the tumor cell PTEN gene expression and effectively inhibited tumor progression by inducing tumor cell immunogenic cell death (ICD) and increasing immune cell infiltration. Moreover, we developed an anti-PD-1 loaded phospholipid-based phase separation gel (PPSG), which formed an in situ depot and sustainably release anti-PD-1 drugs within 42 days in vivo. In order to effectively inhibit the recurrence of melanoma, we further applied a triple therapy based on AAV6-PTEN, PPSG^@anti-PD-1 and CpG, and showed that this triple therapy strategy enhanced the synergistic antitumor immune effect and also induced robust immune memory, which completely rejected tumor recurrence. We anticipate that this triple therapy could be used as a new tumor combination therapy with stronger immune activation capacity and tumor inhibition efficacy.

Background::It is of great clinical significance to further explore new strategies and potential combined therapeutic targets for gastric cancer. This study aimed to investigate the synthetic lethal effect of RBBP8 molecular intervention combined with a poly ADP ribose polymerase (PARP) inhibitor in non-BRCA mutant gastric cancer and clarify the mechanism by which RBBP8 regulates homologous recombination repair.Methods::The role of RBBP8 in DNA damage repair was observed using bioinformatic analysis, western blot analysis, and immunofluorescence. The synthetic lethal effect was verified using 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt (MTS)and flow cytometry apoptosis experiments.Results::Among the patients with gastric cancer treated with chemotherapy, the prognosis of patients with high RBBP8 expression levels was worse (homologous recombination [HR]= 1.54, p= 0.028). RBBP8 knockdown induced DNA damage and had a synergistic effect with PARP inhibitor treatment on cell viability inhibition and cell apoptosis in AGS (generic code for human gastric adenocarcinoma cells) ( t= 11.154, p < 0.001) and N87 ( t= 6.362, p < 0.001) cells. RBBP8 knockdown inhibited RAD51 activation and DNA terminal excision in homologous recombination repair. Conclusion::RBBP8 is involved in homologous recombination repair, and molecular intervention into RBBP8 could achieve a synthetic lethal effect with PARP inhibitor treatment in gastric cancer cells.

Background::It is of great clinical significance to further explore new strategies and potential combined therapeutic targets for gastric cancer. This study aimed to investigate the synthetic lethal effect of RBBP8 molecular intervention combined with a poly ADP ribose polymerase (PARP) inhibitor in non-BRCA mutant gastric cancer and clarify the mechanism by which RBBP8 regulates homologous recombination repair.Methods::The role of RBBP8 in DNA damage repair was observed using bioinformatic analysis, western blot analysis, and immunofluorescence. The synthetic lethal effect was verified using 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt (MTS)and flow cytometry apoptosis experiments.Results::Among the patients with gastric cancer treated with chemotherapy, the prognosis of patients with high RBBP8 expression levels was worse (homologous recombination [HR]= 1.54, p= 0.028). RBBP8 knockdown induced DNA damage and had a synergistic effect with PARP inhibitor treatment on cell viability inhibition and cell apoptosis in AGS (generic code for human gastric adenocarcinoma cells) ( t= 11.154, p < 0.001) and N87 ( t= 6.362, p < 0.001) cells. RBBP8 knockdown inhibited RAD51 activation and DNA terminal excision in homologous recombination repair. Conclusion::RBBP8 is involved in homologous recombination repair, and molecular intervention into RBBP8 could achieve a synthetic lethal effect with PARP inhibitor treatment in gastric cancer cells.