Foxp3+CD4+ regulatory T cells (Tregs) are crucial in maintaining self-tolerance and limiting immune responses to pathogens. Shifting the sensitive balance between Tregs and effector T cells requires extensive knowledge of the homeostatic properties of the different T cell populations. For the investigation of Treg homeostatic expansion, we introduce in this study novel BAC transgenic mice, designated Foxp3.LuciDTR, coexpressing enhanced GFP, luciferase for bioluminescence imaging of Tregs, and the diphtheria toxin receptor (DTR) for specific ablation of Tregs. Of several founder lines, Foxp3.LuciDTR-4 mice displayed ∼95% Treg depletion following injection of DT, resulting in activation of conventional CD4+ T cells, probably due to lack of control by Tregs. In contrast, Foxp3.LuciDTR-3 mice displayed only ∼70% Treg depletion without concomitant activation of CD4+ T cells and represented, therefore, a suitable model to study Treg homeostasis in an environment where other T cell populations were not altered. After depletion, the Treg compartment recovered to its original size in ∼2 wk. This recovery was mediated in a thymus-independent fashion by homeostatic proliferation of the surviving, nondepleted Tregs. The proliferating Tregs acquired an activated phenotype and maintained their suppressive capacity. Studies involving DT-mediated depletion of dendritic cells in CD11c.DOG mice showed that dendritic cells were required for optimal Treg homeostasis. In addition, IL-2 was identified as an essential factor for homeostatic recovery of the Treg compartment. These results show that Treg homeostasis is specifically regulated by the size of the Treg compartment and is independent of proliferation of conventional T cells.

During the coronavirus disease 2019 (COVID-19) outbreak in China, fear about COVID-19, together with worry about progression of cancer, caused strong emotional stress in patients with cancer. We evaluated patientreported outcome in 658 patients with breast cancer (BC) and survivors recruited from multiple BC centers in Hubei Province using 4 standardized assessment scales. Multivariable logistic regression analysis was used to identify potential affecting factors on mental health outcomes. High rates of anxiety, depression, distress, and insomnia were observed in patients with BC during the COVID-19 outbreak. Based on our results, living in Wuhan, poor general condition by self-identification, shorter duration after BC diagnosis, aggressive BC molecular subtypes, metastatic BC clinical stage, treatment discontinuation, central venous catheter flushing delay, or close contact with patients with COVID-19 are associated risk factors for poorer psychological status. Special attention should be paid to the psychological status of patients with BC, especially those with poor general condition, treatment discontinuation, aggressive molecular subtypes, and metastatic BC. Introduction: We aimed to analyze the psychological status in patients with breast cancer (BC) in the epicenter of the coronavirus disease 2019 (COVID-19) pandemic. Patients and Methods: A total of 658 individuals were recruited from multiple BC centers in Hubei Province. Online questionnaires were conducted, and these included demographic information, clinical features, and 4 patient-reported outcome scales (Generalized Anxiety Disorder Questionnaire [GAD-7], Patient Health Questionnaire [PHQ-9], Insomnia Severity Index [ISI], and Impact of Events Scale-Revised [IES-R]). Multivariable logistic regression analysis was designed to identify potential factors on mental health outcomes. Results: Questionnaires were collected from February 16, 2020 to February 19, 2020, the peak time point of the COVID-19 outbreak in China. Of patients with BC, 46.2% had to modify planned necessary anti-cancer treatment during the outbreak. Severe anxiety and severe depression were reported by 8.9% and 9.3% of patients, respectively.

A s the most common cancer among women worldwide, breast cancer poses a great challenge to public health on a global scale (1). Identification of the presence of lymph node metastasis is pivotal for the pathologic staging, prognosis, and guidance of treatment in patients with breast cancer (2). Although several histopathologic findings, such as vascular and lymphatic invasion, epithelial hyperplasia, and necrosis, are associated with a higher risk for lymph node metastasis, they are available only postoperatively (3). The preoperative prediction of lymph node metastasis can provide valuable information for determining adjuvant therapy and developing surgical plans, thereby facilitating pretreatment decisions.Preoperative imaging assessment is of great value because of its convenient, comprehensive, and noninvasive properties. US plays a crucial role in detecting breast cancer and predicting lymph node metastasis (4). Most patients with early stage breast cancer who have clinically negative lymph nodes have no suspicious signs at either physical examination or imaging. Although radiologists often cannot find any signs of metastasis on US images of clinically negative lymph nodes, axillary lymph node metastasis is detected with sentinel lymph node biopsy in 15%-20% of patients (5). Several studies have found that numerous breast US characteristics are associated with lymph node metastasis. The distance

There are a number of factors that hamper immunotherapy of cancer. For example, tumors exhibit an aberrant vasculature that appears to form a barrier against T-cell infiltration. Another major obstacle is created by Treg. So far, conventional depletion of Treg with anti-CD25 antibodies, which eliminate only 70% of Treg, has failed to significantly reduce the growth of established tumors. Using Foxp3.LuciDTR-4 mice, we show here that 90-95% Treg depletion resulted in complete regression of large established tumors, whereas 70% depletion was ineffective. The extensive Treg depletion induced a number of processes that are critical for tumor rejection, including activation of tumorspecific CD81 T cells and enhanced infiltration of these cells into the tumor. The precise mechanism of enhanced infiltration is not known, but normalization of the tumor vasculature is assumed to assist infiltration. Indeed, we observed that 90% Treg depletion caused normalization of the tumor vasculature as indicated by a reduction in leakiness and the number of dilated vessels. These results suggest that for clinical immunotherapy of cancer, it would be desirable to have reagents that allow high-level depletion of Treg, which, in conjunction with treatment modalities such as vaccination, may concomitantly increase T-cell activation and infiltration.Key words: Immunotherapy . Treg . Tumor immunology Supporting Information available online IntroductionGrowing evidence from various malignant entities including melanoma, colorectal, ovarian, cervical, hepatocellular, gastric and urothelial carcinoma indicates that an increased number of tumor infiltrating lymphocytes correlates with favorable disease course and longer survival [1][2][3][4][5][6][7][8][9]. A major obstacle for infiltration appears to be the tumor endothelial barrier. Neoangiogenesis of blood vessels is necessary for tumor survival, but the resulting tumor vasculature usually displays an abnormal architecture characterized by dilated and fragile vessels resulting in leakiness, hypoxia, acidosis and high interstitial pressure [10,11]. Interestingly, normalization of the tumor vasculature by experimental manipulation leads to increased infiltration and tumor destruction [12]. Thus, normalization of the tumor vasculature emerges as a novel and promising approach for immunotherapy. Several studies have attempted to interfere with the growth of mouse transplantation tumors by PC-61 monotherapy. Usually an effect on tumor growth has been observed only in prophylactic settings when Treg were depleted before or no later than 2 days after tumor inoculation, whereas PC-61-mediated Treg depletion in animals carrying established tumors had generally no or only little effect on tumor growth [25][26][27][28]. Significant therapeutic success was only observed when complex combinatorial approaches were applied such as total body irradiation combined with adoptive T-cell transfer and vaccination [29], and additional treatment with anti-PDL1 antibody [30], but usually mice were treated when tu...

Microfluidic chips with nano-scale structures have shown great potential, but the fabrication and cost issues restrict their application. Herein, we propose a conceptually new "DNA nanolithography in a microfluidic chip" by using sub-10 nm three-dimensional DNA structures (TDNs) as frameworks with a pendant aptamer at the top vertex (ApTDN-Chip). The nano-scale framework ensures that the aptamer is in a highly ordered upright orientation, avoiding the undesired orientation or crowding effects caused by conventional microfluidic interface fabrication processes. Compared with a monovalent aptamer modified chip, the capture efficiency of ApTDN-Chip was enhanced nearly 60 % due to the highly precise dimension and rigid framework of TDNs. In addition, the scaffolds make DNase I more accessible to the aptamer with up to 83 % release efficiency and 91 % cell viability, which is fully compatible with downstream molecular analysis. Overall, this strategy provides a novel perspective on engineering nano-scaffolds to achieve a more ordered nanotopography of microfluidic chips.

Aflatoxin B1 (AFB1), as the secondary metabolite of molds, is the most predominant and toxic mycotoxin that seriously threatens the health of humans and animals. In this work, an AFB1-responsive hydrogel was synthesized for highly sensitive and portable detection of AFB1. The AFB1-responsive hydrogel was prepared using an AFB1 aptamer and its two short complementary DNA strands as cross-linkers. For visual detection of AFB1, the hydrogel is preloaded with gold nanoparticles (AuNPs). Upon introduction of AFB1, the AFB1 aptamer binds with AFB1, leading to the disruption of the hydrogel and release of the AuNPs with a distinct color change of the supernatant from colorless to red. In order to lower the detection limit and extend the method to quantitative analysis, a distance-readout volumetric bar chart chip (V-chip) was combined with an AFB1-responsive hydrogel preloaded with platinum nanoparticles (PtNPs). In the presence of AFB1, the hydrogel collapses and releases PtNPs which can catalyze the decomposition of H2O2 to generate O2. The increasing gas pressure moves a red ink bar in the V-chip and provides a quantitative relationship between the distance and the concentration of AFB1. The method was applied for detection of AFB1 in beer, with a detection limit of 1.77 nM (0.55 ppb) where an immunoaffinity column (IAC) of AFB1 was used to cleanup and pre-concentrate the sample, which satisfies the testing requirement of 2.0 ppb set by the European Union. The combination of an AFB1-responsive hydrogel with a distance-based readout V-chip offers a user-friendly POCT device, which has great potential for rapid, portable, selective, and quantitative detection of AFB1 in real samples to ensure food safety and avoid subsequent economic losses.

Background: Cancer patients had been profoundly affected by the outbreak of COVID-19 especially after quarantine restrictions in China. We aimed to explore the treatment changes and delays of early breast cancer (EBC) during the first quarter of 2020. Methods: We did this retrospective, multicentre, cohort study at 97 cancer centres in China. EBC patients who received treatment regardless of preoperative therapy, surgery or postoperative therapy during first quarter of 2020 were included. Findings: 8397 patients were eligible with a median age of 50 (IQR 43À56). 0¢2% (15/8397) of EBC patients were confirmed as COVID-19 infection. Only 5¢2% of breast cancer diagnosis occurred after quarantine in Hubei compared with 15¢3% in other provinces (OR= 0¢30, 95%CI 0¢24À0¢38). postoperative endocrine

IntroductionPrevious studies have indicated that the small leucine-rich proteoglycan (SLR) osteoglycin (OGN) is downregulated in various cancers, including squamous cervical carcinoma, gastric cancer, and colorectal adenoma, indicating that OGN is a putative tumor suppressor. However, its exact role in the pathology of human cancers, especially breast cancer (BC), is not clear.MethodsThe expression of OGN in BC tissues was examined using qRT-PCR. Online databases were employed to analyze the correlation between OGN expression and clinicopathological characteristics. CCK-8 assay, colony formation assay, transwell migration and invasion assays were applied to detect cell proliferation, colony formation, migration and invasion of BC cells, respectively. Xenograft tumor models were constructed to explore the role of OGN on tumor growth in vivo.ResultsOGN expression was reduced in 24 paired BC samples compared with normal tissue. Decreased expression of OGN was correlated with greater pathological grade, a more aggressive tumor subtype, and poor overall survival. In vitro experiments showed that OGN overexpressed by plasmid transfection significantly inhibited cell proliferation, colony formation, migration, and invasion of BC cell lines. In xenograft tumor models, overexpression of OGN repressed the growth of MCF-7 cells in vivo and alleviated the compression of the tumor on surrounding structures. We also observed that OGN expression reversed EMT via repressing the PI3K/Akt/mTOR pathway.ConclusionThis study revealed that OGN could function as a tumor suppressor during breast carcinogenesis, and we contribute new evidence to the body of research on the SLRP family.