Accurate prediction of peritoneal metastasis for gastric cancer (GC) with serosal invasion is crucial in clinic. The presence of collagen in the tumour microenvironment affects the metastasis of cancer cells. Herein, we propose a collagen signature, which is composed of multiple collagen features in the tumour microenvironment of the serosa derived from multiphoton imaging, to describe the extent of collagen alterations. We find that a high collagen signature is significantly associated with a high risk of peritoneal metastasis (P < 0.001). A competing-risk nomogram including the collagen signature, tumour size, tumour differentiation status and lymph node metastasis is constructed. The nomogram demonstrates satisfactory discrimination and calibration. Thus, the collagen signature in the tumour microenvironment of the gastric serosa is associated with peritoneal metastasis in GC with serosal invasion, and the nomogram can be conveniently used to individually predict the risk of peritoneal metastasis in GC with serosal invasion after radical surgery.

AimsTo investigate the effect of survivin (SVV)-engineered mesenchymal stem cells (MSCs) on post-infarction cardiac performance and remodelling in rats. Methods and resultsMesenchymal stem cells from male Sprague-Dawley rat bone marrow were infected with the self-inactive lentiviral vector GFP-wre-CMV/LTR and Flap-Ubiqutin promoter (GCFU) carrying green fluorescent protein (GFP) gene and SVV recombinant vector (GCFU-SVV). In vitro, modification with SVV increased the secretion of vascular endothelial growth factor (VEGF) by 1.28-fold under hypoxic conditions. In vivo, after permanent left anterior descending artery occlusion, rats were randomized (n ¼ 18 per group) to receive intra-myocardial injections of 100 mL of phosphatebuffered saline without cells (group vehicle) or containing 2 million MSC GFP (group MSC GFP ) or MSC SVV (group MSC SVV ) cells. Cellular survival assessed by reverse transcriptase-polymerase chain reaction for GFP in the MSC SVV group was 2.5-fold higher at 7 days and 4.3-fold higher at 28 days after transplantation than in the MSC GFP group. When compared with transplantation with MSC GFP , transplantation with MSC SVV further upregulated VEGF expression at 7 and 28 days after myocardial infarction (MI), increased capillary density by 38%, reduced the infarct size by 12.7%, significantly inhibited collagen deposition, and further improved cardiac function at 28 days after MI. ConclusionTransplantation with SVV-engineered MSCs by lentiviral vector leads to better prognosis for MI by enhancing cellular survival.--

The notion of personalized medicine demands proper prognostic biomarkers to guide the optimal therapy for an invasive breast cancer patient. However, various risk prediction models based on conventional clinicopathological factors and emergent molecular assays have been frequently limited by either a low strength of prognosis or restricted applicability to specific types of patients. Therefore, there is a critical need to develop a strong and general prognosticator. Methods: We observed five large-scale tumor-associated collagen signatures (TACS4-8) obtained by multiphoton microscopy at the invasion front of the breast primary tumor, which contrasted with the three tumor-associated collagen signatures (TACS1-3) discovered by Keely and coworkers at a smaller scale. Highly concordant TACS1-8 classifications were obtained by three independent observers. Using the ridge regression analysis, we obtained a TACS-score for each patient based on the combined TACS1-8 and established a risk prediction model based on the TACS-score. In a blind fashion, consistent retrospective prognosis was obtained from 995 breast cancer patients in both a training cohort ( n = 431) and an internal validation cohort ( n = 300) collected from one clinical center, and in an external validation cohort ( n = 264) collected from a different clinical center. Results: TACS1-8 model alone competed favorably with all reported models in predicting disease-free survival (AUC: 0.838, [0.800-0.872]; 0.827, [0.779-0.868]; 0.807, [0.754-0.853] in the three cohorts) and stratifying low- and high-risk patients (HR 7.032, [4.869-10.158]; 6.846, [4.370-10.726], 4.423, [2.917-6.708]). The combination of these factors with the TACS-score into a nomogram model further improved the prognosis (AUC: 0.865, [0.829-0.896]; 0.861, [0.816-0.898]; 0.854, [0.805-0.894]; HR 7.882, [5.487-11.323]; 9.176, [5.683-14.816], and 5.548, [3.705-8.307]). The nomogram identified 72 of 357 (~20%) patients with unsuccessful 5-year disease-free survival that might have been undertreated postoperatively. Conclusions: The risk prediction model based on TACS1-8 considerably outperforms the contextual clinical model and may thus convince pathologists to pursue a TACS-based breast cancer prognosis. Our methodology identifies a significant portion of patients susceptible to undertreatment (high-risk patients), in contrast to the multigene assays that often strive to mitigate overtreatment. The compatibility of our methodology with standard histology using traditional (non-tissue-microarray) formalin-fixed paraffin-embedded (FFPE) tissue sections could simplify subsequent clinical translation.

Liver surface is covered by a collagenous layer called the Glisson’s capsule. The structure of the Glisson’s capsule is barely seen in the biopsy samples for histology assessment, thus the changes of the collagen network from the Glisson’s capsule during the liver disease progression are not well studied. In this report, we investigated whether non-linear optical imaging of the Glisson’s capsule at liver surface would yield sufficient information to allow quantitative staging of liver fibrosis. In contrast to conventional tissue sections whereby tissues are cut perpendicular to the liver surface and interior information from the liver biopsy samples were used, we have established a capsule index based on significant parameters extracted from the second harmonic generation (SHG) microscopy images of capsule collagen from anterior surface of rat livers. Thioacetamide (TAA) induced liver fibrosis animal models was used in this study. The capsule index is capable of differentiating different fibrosis stages, with area under receiver operating characteristics curve (AUC) up to 0.91, making it possible to quantitatively stage liver fibrosis via liver surface imaging potentially with endomicroscopy.

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Abstract. Intrinsic two-photon excited fluorescence ͑TPEF͒ and second-harmonic generation ͑SHG͒ signals are shown to differentiate between normal and neoplastic human esophageal stroma. It was found that TPEF and SHG signals from normal and neoplastic stroma exhibit different organization features, providing quantitative information about the biomorphology and biochemistry of tissue. By comparing normal with neoplastic stroma, there were significant differences in collagen-related changes, elastinrelated changes, and alteration in proportions of matrix molecules, giving insight into the stromal changes associated with cancer progression and providing substantial potential to be applied in vivo to the clinical diagnosis of epithelial precancers and cancers. A number of optical spectroscopic and imaging techniques have widely been used for the in vivo, real-time detection of epithelial precancers and cancers.1 However, most studies focus only on the epithelium to analyze morphological, structural, and architectural changes that accompany development of epithelial precancers.2 Recently, it has been recognized that stromal biology is also altered significantly with various pathological processes.2 However, there is a lack of utilizing the alterations in the stroma as an intrinsic indicator of disease states, which can extract quantitative information about the biomorphology and biochemistry and can provide a new means to improve early detection of neoplastic changes.Multiphoton microscopy has several advantages over traditional confocal microscopy, providing high-resolution images at increased imaging depths, minimal out-of-plane absorption, and inherent optical sectioning.3 It is well suited for the observation of unstained samples based on intrinsic sources of nonlinear signals, 4 such as two-photon excited fluorescence ͑TPEF͒ and second-harmonic generation ͑SHG͒. It also has been used for in vivo cancer imaging. 3Our primary goals in this study are to investigate the intrinsic TPEF and SHG signals in epithelial stroma and to extract biomorphologic and biochemical features for potential tissue diagnosis.The multiphoton system used in this study has been described previously. 4 Briefly, it is an Axiovert 200 microscope ͑Zeiss LSM 510 META͒ equipped with a mode-locked femtosecond Ti:sapphire laser ͑Coherent Mira 900-F͒. The polarization of laser light is the linear polarization. To suppress TPEF signals from collagen, the excitation wavelength at 850 nm was used in this work. 4 An oil immersion objective ͑ϫ63 and NA= 1.4͒ was employed. The META detector was used to detect all signals in backscattered geometry. The average laser power at the sample was 5 mW. In this study, all images were 512ϫ 512 pixels. The images were obtained at 2.56 s per pixel. In this work, considering that the surface of the sections is not absolutely flat and that the signals in the surface of sections is not absolutely maximal, we defined a depth of 0 m, referring to the position where the multiphoton signals of reflection from the interfac...

Multiphoton microscopic imaging of collagen plays an important role in noninvasive diagnoses of human tissue. In this study, two-photon fluorescence and second-harmonic generation (SHG) imaging of collagen in human skin dermis and submucosa of colon and stomach tissues were investigated based on multiphoton microscopy (MPM). Our results show that multiphoton microscopic image of collagen bundles exhibits apparently different pattern in human tissues. The collagen bundles can simultaneously reveal its SHG and two-photon excited fluorescence images in the submucosa of colon and stomach, whereas it solely emit SHG signal in skin dermis. The intensity spectral information from tissues further demonstrated the above results. This indicates that collagen bundles have completely different space arrangement in these tissues. Our experimental results bring more detailed information of collagen for the application of MPM in human noninvasive imaging.

Bile canaliculi expand and contract in response to the amount of secreted bile, and resistance from the surrounding actin bundles. Further expansion due to bile duct blockade leads to the formation of inward blebs, which carry away excess bile to prevent bile build up in the canaliculi.