The value of narrow-band imaging bronchoscopy in diagnosing central lung cancer.

Aims: This research aimed to study the value of narrow-band imaging(NBI) in the diagnosis of central lung cancer. Materials and methods: This study included 916 patients with clinical suspected of central lung cancer or follow-up of patients after curative lung cancer surgery. All of the patients were examined by Olympus Evis Lucera electronic bronchoscope system, any sites that were abnormal when viewed by white-light bronchoscopy (WLB) or NBI were biopsied, four to six biopsies were taken at each site of the abnormal region visualized as lesions, we record the endoscopic features of NBI and compared with histopathology results, to evaluate the diagnostic value of NBI for central lung cancer and the relationship between vascular patterns of NBI and histological types of lung cancer, and try to establish a multinomial logistic regression model for predicting the histological types of lung cancer. The biopsy specimens were examined by CD34 antibody through immunohistochemistry (IHC) method, CD34 marked microvessel density(MVD), compared the number of microvessels between benign and malignant diseases and the number between different histological types of lung cancer, to verify the results of NBI. Results: NBI provided high sensitivity (91.7%), specificity (84.9%), positive predictive value (97.6%), negative predictive value (61.5%), and agreement rate (90.7%). The predominant vascular patterns in the well-defined histological types of lung cancer were dotted blood vessels (121 patients), tortuous blood vessels (248 patients), and abrupt-ending blood vessels (227 patients). Logistic regression analysis of the results showed that smoking status of the patient, combined with vascular patterns under NBI, and age partly affect the histological types of lung cancer. Conclusions: NBI is highly accurate for the diagnosis of central lung cancer.

Combined PARP1-Targeted Nuclear Contrast and Reflectance Contrast Enhance Confocal Microscopic Detection of Basal Cell Carcinoma.

Reflectance confocal microscopy (RCM) with endogenous backscattered contrast can noninvasively image basal cell carcinomas (BCCs) in skin. However, BCCs present with high nuclear density, and the relatively weak backscattering from nuclei imposes a fundamental limit on contrast, detectability, and diagnostic accuracy. We investigated PARPi-FL, an exogenous nuclear poly(adenosine diphosphate ribose) polymerase (PARP1)-targeted fluorescent contrast agent, and fluorescence confocal microscopy toward improving BCC diagnosis. Methods: We tested PARP1 expression in 95 BCC tissues using immunohistochemistry, followed by PARPi-FL staining in 32 fresh surgical BCC specimens. The diagnostic accuracy of PARPi-FL contrast was evaluated in 83 surgical specimens. The optimal parameters for permeability of PARPi-FL through intact skin was tested ex vivo on 5 human skin specimens and in vivo in 3 adult Yorkshire pigs. Results: We found significantly higher PARP1 expression and PARPi-FL binding in BCCs than in normal skin structures. Blinded reading of RCM-and-fluorescence confocal microscopy images by 2 experts demonstrated a higher diagnostic accuracy for BCCs with combined fluorescence and reflectance contrast than for RCM alone. Optimal parameters (time and concentration) for PARPi-FL transepidermal permeation through intact skin were successfully determined. Conclusion: Combined fluorescence and reflectance contrast may improve noninvasive BCC diagnosis with confocal microscopy.

Sterile cerebrospinal fluid ascites, hydrothorax and hydrocele as a complication of ventriculoperitoneal shunting in an elderly patient.

An 89-year-old man with a history of multiple abdominal surgeries and ventriculoperitoneal (VP) shunt placement for normal pressure hydrocephalus presented for intractable abdominal bloating and scrotal swelling, for which imaging revealed massive ascites, bilateral hydrocele and small bilateral pleural effusions. Cardiac, hepatic and renal workup were insignificant. Culture and cytology of ascitic fluid were negative for infection or malignancy. Aetiology of the ascites as secondary to Cerebrospinal fluid (CSF) from the VP shunt was confirmed via ligation of the shunt. Sterile CSF ascites, hydrothorax and hydrocele are rare complications of VP shunt for hydrocephalus and are mostly presented in paediatric patients. We report the first known case of concurrent CSF ascites, hydrothorax and hydrocele in an elderly patient. We examine the difficulty of shunt replacement as a diagnostic and treatment modality in this age group and propose the use of reversible shunt ligation as a diagnostic modality.

Fibril bending stiffness of 3D collagen matrices instructs spreading and clustering of invasive and non-invasive breast cancer cells.

Extracellular matrix stiffening of breast tissues has been clinically correlated with malignant transformation and poor prognosis. An increase of collagen fibril diameter and lysyl-oxidase mediated crosslinking has been observed in advanced tumor stages. Many current reports suggest that the local mechanical properties of single fibrillar components dominantly regulate cancer cell behavior. Here, we demonstrate by an independent control of fibril diameter and intrafibrillar crosslinking of three-dimensional (3D) collagen matrices that fibril bending stiffness instructs cell behavior of invasive and non-invasive breast cancer cells. Two types of collagen matrices with fibril diameter of either 650 nm or 800 nm at a similar pore size of 10 µm were reconstituted and further modified with the zero-length crosslinker 1-ethyl-3-(3-dimethyl aminopropyl)-carbodiimide (EDC) at concentrations of 0, 20, 100 and 500 mM. This approach yields two sets of collagen matrices with overlapping variation of matrix elasticity. With these matrices we could prove the common assumption that matrix elasticity of collagen networks is bending dominated with a linear dependence on fibril bending stiffness. We derive that the measured variation of matrix elasticity is directly correlated to the variation of fibril bending stiffness, being independently controlled either by fibril diameter or by intrafibrillar crosslinking. We use these defined matrices to demonstrate that the adjustment of fibril bending stiffness allows to instruct the behavior of two different breast cancer cell lines, invasive MDA-MB-231 (human breast carcinoma) and non-invasive MCF-7 cells (human breast adenocarcinoma). Invasiveness and spreading of invasive MDA-MB-231 cells as well as clustering of non-invasive MCF-7 cells is thereby investigated over a broad parameter range. Our results demonstrate and quantify the direct dependence of cancer cell phenotypes on the matrix mechanical properties on the scale of single fibrils.

Fibroblast fate regulation by time dependent TGF-ß1 and IL-10 stimulation in biomimetic 3D matrices.

The presentation of TGF-ß1 during the early stage of wound healing is a prerequisite for extracellular matrix (ECM) synthesis and remodeling by activated fibroblasts, called myofibroblasts. At later stages, clearance of myofibroblasts is needed to avoid overshooting ECM production. Apoptosis of myofibroblasts and the macrophage-released anti-inflammatory cytokine IL-10 are controversially discussed as regulating cues in this context. To reveal the regulating cues, defined biomaterial scaffolds are needed to conduct in-depth in vitro studies in a physiologically relevant context. In this work, we used an in vitro biomimetic wound healing model. It consists of a 3D fibrillar matrix from collagen I and fibronectin and different temporal stimuli by TGF-ß1 and IL-10. Human dermal fibroblast behavior was investigated in terms of myofibroblast differentiation (αSMA expression), matrix remodeling, proliferation and migration in the permanent or sequential presence of TGF-ß1 and IL-10 over 4 days. We could show that removal of TGF-ß1 after initial stimulation resulted in an increase of apoptosis of myofibroblasts. In contrast, TGF-ß1 stimulation followed by IL-10 treatment did not result in increased cell apoptosis but instead led to a significant increase of cell motility and reduction of myofibroblasts. The findings suggest that myofibroblasts are a transiently "activated" fibroblastic phenotype and can be de-differentiated to fibroblasts in the presence of IL-10. Overall, our 3D ECM model allows mimicking the early and late stages of wound healing and highlights the temporal sequence of TGF-ß1 and IL-10 as an important cue for completion of tissue formation and maintenance of tissue homeostasis.

CD24 Expression and differential resistance to chemotherapy in triple-negative breast cancer.

Breast cancer (BC) is a leading cause of cancer-related death in women. Adjuvant systemic chemotherapies are effective in reducing risks of recurrence and have contributed to reduced BC mortality. Although targeted adjuvant treatments determined by biomarkers for endocrine and HER2-directed therapies are largely successful, predicting clinical benefit from chemotherapy is more challenging. Drug resistance is a major reason for treatment failures. Efforts are ongoing to find biomarkers to select patients most likely to benefit from chemotherapy. Importantly, cell surface biomarkers CD44+/CD24- are linked to drug resistance in some reports, yet underlying mechanisms are largely unknown. This study focused on the potential role of CD24 expression in resistance to either docetaxel or doxorubicin in part by the use of triple-negative BC (TNBC) tissue microarrays. In vitro assays were also done to assess changes in CD24 expression and differential drug susceptibility after chemotherapy. Further, mouse tumor xenograft studies were done to confirm in vitro findings. Overall, the results show that patients with CD24-positive TNBC had significantly worse overall survival and disease-free survival after taxane-based treatment. Also, in vitro cell studies show that CD44+/CD24+/high cells are more resistant to docetaxel, while CD44+/CD24-/low cells are resistant to doxorubicin. Both in vitro and in vivo studies show that cells with CD24-knockdown are more sensitive to docetaxel, while CD24-overexpressing cells are more sensitive to doxorubicin. Further, mechanistic studies indicate that Bcl-2 and TGF-ßR1 signaling via ATM-NDRG2 pathways regulate CD24. Hence, CD24 may be a biomarker to select chemotherapeutics and a target to overcome TNBC drug resistance.