A comprehensive profile of TCF1+ progenitor and TCF1- terminally exhausted PD-1+CD8+ T cells in head and neck squamous cell carcinoma: implications for prognosis and immunotherapy / 国际口腔科学杂志·英文版

The heterogeneity of exhausted T cells (Tex) is a critical determinant of immune checkpoint blockade therapy efficacy. However, few studies have explored exhausted T cell subpopulations in human cancers. In the present study, we examined samples from two cohorts of 175 patients with head and neck squamous cell cancer (HNSCC) by multiplex immunohistochemistry (mIHC) to investigate two subsets of Tex, CD8+PD1+TCF1+ progenitor exhausted T cells (TCF1+Texprog) and CD8+PD1+TCF1- terminally exhausted T cells (TCF1-Texterm). Moreover, fresh tumor samples from 34 patients with HNSCC were examined by flow cytometry and immunohistochemistry to further investigate their properties and cytotoxic capabilities and their correlation with regulatory T cells (Tregs) in the tumor immune microenvironment (TIME). mIHC and flow cytometry analysis showed that TCF1-Texterm represented a greater proportion of CD8+PD1+Tex than TCF1+Texprog in most patients. TCF1+Texprog produced abundant TNFα, while TCF1-Texterm expressed higher levels of CD103, TIM-3, CTLA-4, and TIGIT. TCF1-Texterm exhibited a polyfunctional TNFα+GZMB+IFNγ+ phenotype; and were associated with better overall survival and recurrence-free survival. The results also indicated that larger proportions of TCF1-Texterm were accompanied by an increase in the proportion of Tregs. Therefore, it was concluded that TCF1-Texterm was the major CD8+PD1+Tex subset in the HNSCC TIME and that these cells favor patient survival. A high proportion of TCF1-Texterm was associated with greater Treg abundance.

Bone Morphogenetic Protein Receptor in the Osteogenic Differentiation of Rat Bone Marrow Stromal Cells

PURPOSE: Several signaling pathways have been shown to regulate the lineage commitment and terminal differentiation of bone marrow stromal cells (BMSCs). Bone morphogenetic protein (BMP) signaling has important effects on the process of skeletogenesis. In the present study, we tested the role of bone morphogenetic protein receptor (BMPR) in the osteogenic differentiation of rat bone marrow stromal cells in osteogenic medium (OM) with or without BMP-2. MATERIALS AND METHODS: BMSCs were harvested from rats and cultured in OM containing dexamethasone, beta-glycerophosphate, and ascorbic acid, with or without BMP-2 in order to induce osteogenic differentiation. The alkaline phosphatase (ALP) activity assay and von kossa staining were used to assess the osteogenic differentiation of the BMSCs. BMPR mRNA expression was assessed using reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: The BMSCs that underwent osteogenic differentiation in OM showed a higher level of ALP activity and matrix mineralization. BMP-2 alone induced a low level of ALP activity and matrix mineralization in BMSCs, but enhanced the osteogenic differentiation of BMSCs when combined with OM. The OM significantly induced the expression of type IA receptor of BMPR (BMPRIA) and type II receptor of BMPR (BMPRII) in BMSCs after three days of stimulation, while BMP-2 significantly induced BMPRIA and BMPRII in BMSCs after nine or six days of stimulation, respectively. CONCLUSION: BMSCs commit to osteoblastic differentiation in OM, which is enhanced by BMP-2. In addition, BMP signaling through BMPRIA and BMPRII regulates the osteogenic differentiation of rat BMSCs in OM with or without BMP-2.

Establishment of 3-dimensional finite element model of post-inlay restoration of the first mandibular residual molar crown / 中国组织工程研究

BACKGROUND:During the restoration of residual molar crown, a little part of tooth is still remained commonly. After the restoration, with various forces, stress distribution affects directly the results after restoration. Finite element method is gradually applied in stress analysis on artificial tooth.OBJECTIVE: To establish the three-dimensional (3-D) finite element model of post-inlay restoration of the first residual mandibular molar crown so as to provide experimental data for improving model establishment of complicated teeth and analysis on the property of stress distribution of restoring methods.DESIGN: Repeated observation and measurement were given.SETTING: Department of Stomatology and Department of Radiology of First Hospital affiliated to Sun Yat-sen University;Department of Solid Mechanics,College of Traffics and Communications, South China University of Technology; Department of Restoration of Guanghua College of Stomatology.MATERIALS: The experiment was performed in Department of Solid Mechanics, College of Traffics and Communications of South China University of Technology from November 2003 to December 2004. Six first mandibular molars on the right side with normal morphology in vitro were collected, and Toshiba Xpress/SX spiral CT machine, image photo synthesis software and finite element analysis software ANSYS were applied in the experiment.METHODS: 1 of the 6 first mandibular molars on the right side with normal morphology in vitro was selected for pulpectomy, which was the best in density and near to clinical requirement in morphology. With pulpectomy, the prosthesis of braking-lock post-inlay restoration was prepared. Spiral CT-cross scanning was performed in premolar crown before the restoration, the residual crown with post-inlay in main root canal after restoration and the residual crown with braking-lock second post-inlay restoration. With image photosynthesis software, 3-D digital model of residual tooth and metal part was established and the entire tooth model was prepared after adhesion of two parts. In order to provide better boundary conditions of simulated natural tooth in practice, alveolar bone was considered. Under Mesh order in ANSYS software, automatic mesh generation was performed in the model directly.MAIN OUTCOME MEASURES: Establishment of 3-D finite element models of residual tooth before restoration, post inlay, and alveolar bone and tooth after restoration and the results of mesh generation.RESULTS: By establishing 3-D finite element models of residual tooth before restoration, post inlay, alveolar bone and tooth after restoration and automatic mesh generation, there were altogether 117720 units and 20988nodes. Good geometric similarity presents between the construction model of 3-D finite element model and solid tissue.CONCLUSION: Combination of 3-D finite-element model with spiral Ctcross technology establishes complex dental models, simulates practical conditions authentically and is good in operation.

Therapeutic effect of AdCMVCD/5-FC system and metabolism of 5-FC in the treatment of human tongue squamous cell carcinoma / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To investigate the therapeutic effect and metabolism of 5-fluorocytosine (5-FC) in human tongue squamous carcinoma cells after treatment with adenovirus-mediated cytosine deaminase (AdCMVCD)/5-FC system.</p><p><b>METHODS</b>Human tongue squamous carcinoma cells (Tca8113 cell line) and its xenografts in BALB/c nude mice were treated with AdCMVCD/5-FC system. The killing effect in vitro and bystander effect were detected by microculture tetrazolium (MTT) assay. Tumor inhibition effect and histopathological changes were observed in vivo. High-performance liquid chromatography (HPLC) was performed to determine the metabolism of 5-FC in vitro and in vivo.</p><p><b>RESULTS</b>AdCMVCD/5-FC system had strong killing effect and bystander effect on Tca8113 cells. Both condition media and cell extracts showed two peaks identified as 5-FC and 5-fluorouracil (5-FU) by HPLC and a time-dependent generation of 5-FU and concomitant time-dependent decreases of 5-FC. Compared to the control groups, mice treated with AdCMVCD/5-FC system demonstrated significant tumor regression (P < 0.001); the tumor doubling time prolonged and inhibition rate was 92.62%. There were substantial tumor necrotic areas and infiltrative lymphocytes around necrotic areas in the AdCMVCD/5-FC treated group under light microscope. There was a significantly low concentration of 5-FC and high concentration of 5-FU in tumor tissue, but only 5-FC was found in blood.</p><p><b>CONCLUSION</b>AdCMVCD/5-FC suicide gene system had significant in vitro and in vivo anti-tumor effect on human tongue squamous cell carcinoma due to convert 5-FC into 5-FU.</p>

The therapeutic mechanism of adenovirus mediated CD/5-FC system in the treatment of oral squamous carcinoma cells / 实用口腔医学杂志

Objective: To study the therapeutic mechanism of adenovirus mediated cytosine deaminase /5-fluorocytosine (AdCMVCD/ 5-FC) suicide gene system in the treatment of oral squamous carcinoma cells in vitro. Methods: 3H-thymidine ( 3H-TdR) incorporation assay, flow cytometry (FCM), transmission electron microscope and TUNEL (TdT-mediated dUTP Nick End Labeling)assay were used to detected the changes of Tca8113 cells after the treatment with AdCMVCD/5-FC system. Results: After treatment with CD/5-FC system, 3H-TdR incorporation rate of the cells decreased and significantly decreased between different MOI (multiple of infection) at 5-FC 10 -3 mol/L (P0.05); many apoptotic cells were found under transmission electron microscope and the positive rate of apoptotic cells increased from (4.40?0.87)% (before treatment) to (15.80?1.55)% (P