Development of novel humanized CD19/BAFFR bicistronic chimeric antigen receptor T cells with potent antitumor activity against B-cell lineage neoplasms.

Chimeric antigen receptor T cell (CAR-T) therapy has shown remarkable efficacy in treating advanced B-cell malignancies by targeting CD19, but antigen-negative relapses and immune responses triggered by murine-derived antibodies remain significant challenges, necessitating the development of novel humanized multitarget CAR-T therapies. Here, we engineered a second-generation 4-1BB-CD3ζ-based CAR construct incorporating humanized CD19 single-chain variable fragments (scFvs) and BAFFR single-variable domains on heavy chains (VHHs), also known as nanobodies. The resultant CAR-T cells, with different constructs, were functionally compared both in vitro and in vivo. We found that the optimal tandem and bicistronic (BI) structures retained respective antigen-binding abilities, and both demonstrated specific activation when stimulated with target cells. At the same time, BI CAR-T cells (BI CARs) exhibited stronger tumour-killing ability and better secretion of interleukin-2 and tumour necrosis factor-alpha than single-target CAR-T cells. Additionally, BI CARs showed less exhaustion phenotype upon repeated antigen stimulation and demonstrated more potent and persistent antitumor effects in mouse xenograft models. Overall, we developed a novel humanized CD19/BAFFR bicistronic CAR (BI CAR) based on a combination of scFv and VHH, which showed potent and sustained antitumor ability both in vitro and in vivo, including against tumours with CD19 or BAFFR deficiencies.

High resolution coherent spectral analysis method based on Brillouin scattering in an optical fiber.

High-resolution optical spectral analysis method is of significant importance for those who want to explore the physical world from the frequency domain. Aiming at the resolution degradation of classical coherent optical spectrum analysis (COSA) caused by the mirror phenomenon, this paper modifies the COSA system by introducing two homologous Brillouin scattering beams to serve as the pre-filter and local oscillator (LO), respectively. The central frequencies of the pre-filtered signal and the LO are locked by the Brillouin frequency shifts of those two Brillouin scattering beams. By means of this modification, the pre-filtered signal is located at either the upper-frequency-shifted side or the lower-frequency-shifted sides of the LO but could not exist on both sides of the LO. The proposed method could cancel the mirror phenomenon and thus improve the systematic resolution to 1.3 MHz in theory and 2 MHz in practice.

Investigations of the distant metastatic non-small cell lung cancer without local lymph node involvement: Real world data from a large database.

INTRODUCTION: This study aimed to investigate the presentations and survival outcomes of the distant metastatic non-small cell lung cancer (NSCLC) without lymph node involvement to obtain a clearer picture of this special subgroup of metastatic NSCLC. METHOD: A least absolute shrinkage and selection operator (LASSO) penalized Cox regression analysis was used to select the prognostic variables. A nomogram and corresponding risk-classifying systems were constructed. The C-index and calibration curves were used to evaluate the performance of the model. Overall survival (OS) curves were plotted using the Kaplan-Meier method, and the log-rank test was used to compare OS differences between groups. Propensity score matching (PSM) was performed to reduce bias. RESULT: A total of 12 610 NSCLC patients with M1 category (N0 group: 3045 cases; N1-3 group: 9565 cases) were included. Regarding the N0 group, multivariate analysis demonstrated that age, sex, race, surgery, grade, tumor size, and M category were independent prognostic factors. A nomogram and corresponding risk-classifying systems were formulated. Favorable validation results were obtained from the C-index, calibration curves, and survival comparisons. Survival curves demonstrated that N0 NSCLC patients had better survival than N1-3 NSCLC patients both before and after PSM. Furthermore, the survival of resected N0M1 patients was superior to that of those without surgery. CONCLUSION: In this study, a prognostic nomogram and risk-classifying systems designed for the T1-4N0M1 NSCLC patients showed acceptable performance. Primary lung tumor resection might be a feasible treatment for this population subset. Additionally, we proposed that lymph node stage might have a place in the forthcoming tumor-node-metastasis (TNM) staging proposal for NSCLC patients with M1 category.

miR-124-3p targeting of TGF-ß1 inhibits the proliferation of hypertrophic scar fibroblasts.

BACKGROUND: microRNAs are involved in a variety of physiological and pathophysiological processes, but their role in the pathogenesis of hypertrophic scars (HS) is not fully understood. Transforming growth factor ß1 (TGF-ß1) plays an important role in the genesis and development of HS. OBJECTIVES: In this study, we hypothesized that a post-translational miRNA mechanism regulates the expression of TGF-ß1 in HS fibroblasts (HSFBs) and participates in the development of HS. MATERIAL AND METHODS: Predictions from EBCORI, PicTar and miRBase databases showed that miR-124-3p can target and regulate the expression of TGF-ß1. We collected HS tissue and corresponding normal tissue from 25 patients with HS who had been operated on for the first time. RESULTS: The expression level of miR-124-3p in HS tissue was significantly lower than in normal tissue, while the expression level of TGF-ß1 mRNA was significantly higher than in normal tissue (p < 0.05), showing a negative correlation between them. Results from a luciferase reporter assay showed that miR-124-3p targets the 3'-UTR of TGF-ß1 and inhibits its expression. After miR-124-3p mimics were transfected into HSFBs, the expression of TGF-ß1, α-smooth muscle actin (α-SMA), collagen I, survivin, and Bcl-2 were reduced and the expression of Bax was increased, with significant decreases in DNA synthesis, proliferation and survival. However, after a miR-124-3p inhibitor was transfected into HSFBs, these effects were reversed as the expression of TGF-ß1, α-SMA, collagen I, survivin, and Bcl-2 increased, expression of Bax decreased, and DNA synthesis, proliferation and survival cells increased significantly. CONCLUSIONS: miR-124-3p can inhibit the proliferation of HSFBs by targeting TGF-ß1, and miR-124-3p may thus be a potential therapeutic target in HS.

Serum concentrations of A disintegrin and metalloproteinase 9 (ADAM9) mRNA as a promising novel marker for the detection of pulmonary sarcoidosis.

AIMS: This prospective study investigated the potential role of serum A disintegrin and metalloproteinase 9 (ADAM9) mRNA in the differential diagnosis of pulmonary sarcoidosis from other diseases with similar clinical presentations, such as tuberculosis (TB). METHODS: Serum samples were collected from patients with sarcoidosis, and compared with those from patients with TB and healthy control subjects. Real-time quantitative reverse transcription-polymerase chain reaction was performed to measure the concentration of ADAM9 mRNA relative to the internal control, glyceraldehyde-3-phosphate dehydrogenase. RESULTS: A total of 100 patients with sarcoidosis were compared with 50 patients with TB and 50 healthy control subjects. The serum concentration of ADAM9 mRNA in patients with sarcoidosis was significantly lower than that in healthy control subjects and patients with TB. The serum concentration of ADAM9 mRNA in patients with TB was significantly higher than that in healthy control subjects. CONCLUSION: These data suggest that serum ADAM9 mRNA might be a potential biomarker for the differential diagnosis of pulmonary sarcoidosis.