The 3' Non-Coding Sequence Negatively Regulates PD-L1 Expression, and Its Regulators Are Systematically Identified in Pan-Cancer.

The 3'-untranslated region (3'-UTR) of PD-L1 is significantly longer than the coding sequences (CDSs). However, its role and regulators have been little studied. We deleted whole 3'-UTR region by CRISPR-Cas9. Prognostic analysis was performed using online tools. Immune infiltration analysis was performed using the Timer and Xcell packages. Immunotherapy response prediction and Cox regression was performed using the R software. MicroRNA network analysis was conducted by the Cytoscape software. The level of PD-L1 was significantly and dramatically up-regulated in cells after deleting the 3'-UTR. Additionally, we discovered a panel of 43 RNA-binding proteins (RBPs) whose expression correlates with PD-L1 in the majority of cancer cell lines and tumor tissues. Among these RBPs, PARP14 is widely associated with immune checkpoints, the tumor microenvironment, and immune-infiltrating cells in various cancer types. We also identified 38 microRNAs whose individual expressions are associated with PD-L1 across different cancers. Notably, miR-3139, miR-4761, and miR-15a-5p showed significant associations with PD-L1 in most cancer types. Furthermore, we revealed 21 m6A regulators that strongly correlate with PD-L1. Importantly, by combining the identified RBP and m6A regulators, we established an immune signature consisting of RBMS1, QKI, ZC3HAV1, and RBM38. This signature can be used to predict the responsiveness of cancer patients to immune checkpoint blockade treatment. We demonstrated the critical role of the 3'-UTR in the regulation of PD-L1 and identified a significant number of potential PD-L1 regulators across various types of cancer. The biomarker signature generated from our findings shows promise in predicting patient prognosis. However, further biological investigation is necessary to explore the potential of these PD-L1 regulators.

Perceived Occupational Noise Exposure and Depression in Young Finnish Adults.

We investigated the association between perceived occupational noise exposure and depressive symptoms in young Finnish adults and whether noise sensitivity moderates this association. This study was based on an ongoing longitudinal twin study. We included those who had been working daily (n = 521) or weekly (n = 245) during the past 12 months (mean age 22.4, SD 0.7, 53% female). We asked about occupational noise exposure at age 22 and assessed depressive symptoms using the General Behavior Inventory (GBI) at age 17 and 22. Noise sensitivity and covariates were used in linear regression models. Perceived daily occupational noise exposure was associated, as a statistically independent main effect with depressive symptoms at age 22 (beta 1.19; 95% CI 0.09, 2.29) among all, and separately for females (beta 2.22; 95% CI 0.34, 4.09) but not males (beta 0.22; 95% CI -1.08, 1.52). Noise sensitivity was independently associated with depressive symptoms among all (beta 1.35; 95% CI 0.54, 2.17), and separately for males (beta 1.96; 95% CI 0.68, 3.24) but not females (beta 1.05; 95 % CI -0.04, 2.13). Noise sensitivity was independent of perceived occupational noise exposure. Pre-existing depressive symptoms at age 17 were predictive of perceived occupational noise exposure, suggesting complex interactions of noise and depression.

Nrf2 as a novel diagnostic biomarker for papillary thyroid carcinoma.

Papillary thyroid carcinoma (PTC) is the most common thyroid malignancy. However, it is very difficult to distinguish PTC from benign carcinoma. Thus, specific diagnostic biomarkers are actively pursued. Previous studies observed that Nrf2 was highly expressed in PTC. Based on this research, we hypothesized that Nrf2 may serve as a novel specific diagnostic biomarker. A single-center retrospective study, including 60 patients with PTC and 60 patients with nodular goiter, who underwent thyroidectomy at the Central Theater General Hospital from 2018 to July 2020, was conducted. The clinical data of the patients were collected. Nrf2, BRAF V600E, CK-19, and Gal-3 proteins were compared from paraffin samples of the patients. Through this study, we obtained the following results: i) Nrf2 exhibits high abundance expression in PTC, but not in adjacent to PTC and nodular goiter; increased Nrf2 expression could serve as a valuable biomarker for PTC diagnosis; the sensitivity and specificity for the diagnosis of PTC were 96.70% and 89.40%, respectively. ii) Nrf2 also shows higher expression in PTC with lymph node metastasis, but not adjacent to PTC and nodular goiter, thus the increased Nrf2 expression might serve as a valuable predictor for lymph node metastasis in PTC patients; the sensitivity and specificity for the prediction in lymph node metastasis were 96.00% and 88.57%, respectively; excellent diagnostic agreements were found between Nrf2 and other routine parameters including HO-1, NQO1 and BRAF V600E. iii) The downstream molecular expression of Nrf2 including HO-1 and NQO1 consistently increased. In conclusion, Nrf2 displays a high abundance expression in human PTC, which leads to the higher expression of downstream transcriptional proteins: HO-1 and NQO1. Moreover, Nrf2 can be used as an extra biomarker for differential diagnosis of PTC and a predictive biomarker for lymph node metastasis of PTC.

Serum cystatin C is a potential predictor of short-term mortality and acute kidney injury in acute aortic dissection patients: a retrospective cohort study.

Background: Serum cystatin C concentration is associated with cardiovascular disease. However, the relationship between cystatin C and acute aortic dissection (AAD) remains unclear. In the current study, we aim to evaluate the predictive value of cystatin C in the occurrence of acute kidney injury (AKI) and the prognosis of AAD patients. Methods: The patients with AAD admitted to our hospital from November 2019 through January 2022 were consecutively included in the retrospective cohort study. A complete blood cell count, serum biochemistry tests, including cystatin C and creatinine, in-hospital mortality and the incidence of AKI were recorded. All the patients were categorized into four groups according to the quartile of their serum cystatin C levels. Multivariate logistic and Cox regression analyses were conducted to determine the independent risk factors for the incidence of AKI and the prognosis of AAD patients, respectively. Kaplan-Meier analyses and log-rank tests were used to evaluate differences in survival. Receiver operating characteristic (ROC) curves were used to assess the predictive value of cystatin C for short-term mortality and the incidence of AKI in AAD patients. Results: A total of 357 patients were included in this study. The results showed that the higher the concentration of cystatin C, the higher the level of serum creatinine and the higher the incidence of AKI. Mortality was significantly higher in the group with serum cystatin C levels >1.18 mg/L. Type A AAD, white blood cell count >10×109/L, platelet count <100×109/L, and serum cystatin C concentration >1.18 mg/L [adjusted hazards ratio (HR) =2.405, 95% confidence interval (CI), 1.029-4.063, P=0.041] were independent risk factors for in-hospital mortality. Cystatin C levels >1.18 mg/L remained an independent predictor of AKI in AAD after adjusting for the confounding [odds ratio (OR) 76.489, 95% CI, 25.586-228.660]. The areas under the ROC curves of cystatin C in predicting the mortality and incidence of AKI in AAD patients were 0.655 (95% CI, 0.551-0.760) and 0.807 (95% CI, 0.758-0.856), respectively. Conclusions: In sum, serum cystatin C concentration is a potential predictor of short-term mortality and the incidence of AKI in AAD patients.

Multiscale confocal photoacoustic dermoscopy to evaluate skin health.

Background: Photoacoustic dermoscopy (PAD) is a promising branch of photoacoustic microscopy (PAM) that can provide a range of functional and morphologic information for clinical assessment and diagnosis of dermatological conditions. However, most PAM setups are unsuitable for clinical dermatology because their single-scale mode and narrow frequency band result in insufficient imaging depth or poor spatiotemporal resolution when visualizing the internal texture of the skin. Methods: We developed a multiscale confocal photoacoustic dermoscopy (MC-PAD) with a multifunction opto-sono objective that could achieve high quality dermatological imaging. Using the objective to coordinate the spatial resolution and penetration depth, the MC-PAD was used to visualize pathophysiological biomarkers and vascular morphology from the epidermis (EP) to the dermis, which enabled us to quantify skin abnormalities without using exogenous contrast agents for human skin. Results: The MC-PAD was shown to have the ability to differentiate between different types of cells (such as red blood cells and melanoma cells), image and quantify pigment of the skin, and visualize skin morphology and blood capillary landmarks. The MC-PAD detected a significant difference in the structures of some pigmented and vascular lesions of skin diseases compared with that of healthy skin (P<0.01). The café au lait macule (CALM) skin type was found to have a relatively higher melanin concentration and thicker stratum basale (SB) in the EP than healthy skin. The dermal vascular network of skin that had a port wine stain (PWS) had greater diameters and a denser distribution than healthy skin, as reported in clinical trials. Conclusions: The MC-PAD has a broad range of applications for the diagnosis of human skin diseases and evaluation of the curative effect of treatments, and it can offer new perspectives in biomedical sciences.

Quantitative and Anatomical Imaging of Human Skin by Noninvasive Photoacoustic Dermoscopy.

Imaging plays a vital role in the diagnosis and treatment of skin diseases. However, pure optical imaging technique is limited to the visualization of superficial skin tissues. Ultrasonic imaging technique can detect deep tissues, but it lacks detailed information on microscopic pathological structures. Photoacoustic imaging is an advanced technology that bridges the spatial-resolution gap between optical and ultrasonic techniques, by the modes of optical excitation and acoustic detection. Photoacoustic dermoscopy (PAD), based on photoacoustic technology, can noninvasively obtain high-resolution anatomical structures by endogenous absorbers, such as melanin, hemoglobin, lipids, etc. In the past years, PAD has gradually been developed in clinical dermatology for the diagnosis of melanoma, psoriasis, port-wine stains, dermatitis, skin grafting, and testing the efficacy of cosmetics. This protocol provides detailed procedures for PAD construction, including component selection, equipment setup, and system calibration. A step-by-step guide for human skin imaging is provided as an example application. Image reconstruction and troubleshooting procedures are also elaborated. PAD offers the 3D volumetric images of human skin, and quantitatively analyzes the vascular morphology in the dermis. The protocol will provide clinicians with standardized and reasonable guidance in dermatological imaging.

Quantitative multilayered assessment of skin lightening by photoacoustic microscopy.

BACKGROUND: With the emergence of various new skin-lightening products, there is an urgent need to scientifically evaluate the efficacy and toxicology of these products, and provide scientific guidance for their use based on physiological differences between individuals. Visualized imaging methods and quantitative evaluation criteria play key roles in evaluating the efficacy of skin-lightening products. In order to quantify the changes in the multilayered morphology and endogenous components of human skin before and after the use of lightening products, high-resolution three-dimensional (3D) imaging of human skin is required. METHODS: In this study, photoacoustic microscopy (PAM; SSPM-532, Guangdong Photoacoustic Medical Technology Co., Ltd.) was used to capture the morphological structures of human skin and reveal skin components quantitatively. The efficacy and safety of skin-lightening products were evaluated by measuring skin melanin concentration and observing skin morphology. The melanin concentration in the epidermis was obtained by examining the linear relationship between photoacoustic (PA) signals. Further, the epidermal thickness and the melanin distribution were obtained in the cross-sectional (x-z) and lateral (x-y) images. Finally, the efficacy of skin-lightening products was evaluated according to the concentration and distribution of melanin in the epidermis, and the safety of cosmetics was assessed by observing the vascular morphology in the dermis. RESULTS: PAM noninvasively could assess the multilayered morphological structures of human skin, which allowed for quantification of epidermal thickness and melanin concentration of different skin sites. Based on this, the efficacy and safety of skin-lightening products in multilayer structures were quantitatively evaluated. CONCLUSIONS: As a quantitative imaging method, PAM, has the potential to accurately evaluate the use of skin-lightening products. The method can also be extended to assessments within the larger field of aesthetic medicine.

Quantitative and anatomical imaging of dermal angiopathy by noninvasive photoacoustic microscopic biopsy.

The ability to noninvasively acquire the fine structure of deep tissues is highly valuable but remains a challenge. Here, a photoacoustic microscopic biopsy (PAMB) combined switchable spatial-scale optical excitation with single-element depth-resolved acoustic detection mode was developed, which effectively coordinated the spatial resolution and the penetration depth for visualizations of skin delamination and chromophore structures up to reticular dermis depth, with the lateral resolution from 1.5 to 104 µm and the axial resolution from 34 to 57 µm. The PAMB obtained anatomical imaging of the pigment distribution within the epidermis and the vascular patterns of the deep dermal tissue, enabling quantification of morphological abnormalities of angiopathy without the need for exogenous contrast agents. The features of healthy skin and scar skin, and the abnormal alteration of dermal vasculature in port wine stains (PWS) skin were first precisely displayed by PAMB-shown multi-layered imaging. Moreover, the quantitative vascular parameters evaluation of PWS were carried out by the detailed clinical PAMB data on 174 patients, which reveals distinct differences among different skin types. PAMB captured the PWS changes in capillary-loop depth, diameter, and vascular volume, making it possible to perform an objective clinical evaluation on the severity of PWS. All the results demonstrated the PAMB can provide vascular biopsy and new indexes deep into the dermal skin noninvasively, which should be meaningful to timely evaluate the pathological types and treatment response of skin diseases. This opens up a new perspective for label-free and non-invasive biopsies of dermal angiopathy.

Eremophilane Sesquiterpenes and Benzene Derivatives from the Endophyte Microdiplodia sp. WGHS5.

Three new eremophilane sesquiterpenes phomadecalins G-I (1-3) and two new benzene derivatives microdiplzenes A and B (12 and 13), together with nine known eremophilane sesquiterpenes (4-11 and 14) were isolated from an endophytic fungus, Microdiplodia sp. WGHS5. Their structures were elucidated by the interpretation of HR-ESI-MS and NMR data; meanwhile, the absolute configurations of new compounds were determined on the base of ECD calculations. All compounds were evaluated for the antimicrobial activities and antiproliferative effect on human gastric cancer cell lines (BGC-823).

LINC00941 promotes CRC metastasis through preventing SMAD4 protein degradation and activating the TGF-ß/SMAD2/3 signaling pathway.

ABSTRICT: LINC00941 is a novel lncRNA that has been found to exhibit protumorigenic and prometastatic behaviors during tumorigenesis. However, its role in metastatic CRC remains unknown. We aimed to investigate the functions and mechanisms of LINC00941 in CRC metastasis. LINC00941 was shown to be upregulated in CRC, and upregulated LINC00941 was associated with poor prognosis. Functionally, LINC00941 promoted migratory and invasive capacities and accelerated lung metastasis in nude mice. Mechanistically, LINC00941 activated EMT in CRC cells, as indicated by the increased expression of key molecular markers of cell invasion and metastasis (Vimentin, Fibronectin, and Twist1) and simultaneous decreased expression of the main invasion suppressors E-cadherin and ZO-1. LINC00941 was found to activate EMT by directly binding the SMAD4 protein MH2 domain and competing with ß-TrCP to prevent SMAD4 protein degradation, thus activating the TGF-ß/SMAD2/3 signaling pathway. Our data reveal the essential role of LINC00941 in metastatic CRC via activation of the TGF-ß/SMAD2/3 axis, which provides new insight into the mechanism of metastatic CRC and a novel potential therapeutic target for advanced CRC.

Hyperglycemia associated with lymphopenia and disease severity of COVID-19 in type 2 diabetes mellitus.

BACKGROUND: Coronavirus disease 2019 (COVID-19) has been declared a global pandemic. COVID-19 is more severe in people with diabetes. The identification of risk factors for predicting disease severity in COVID-19 patients with type 2 diabetes mellitus (T2DM) is urgently needed. METHODS: Two hundred and thirty-six patients with COVID-19 were enrolled in our study. The patients were divided into 2 groups: COVID-19 patients with or without T2DM. The patients were further divided into four subgroups according to the severity of COVID-19 as follows: Subgroup A included moderate COVID-19 patients without diabetes, subgroup B included severe COVID-19 patients without diabetes, subgroup C included moderate COVID-19 patients with diabetes, and subgroup D included severe COVID-19 patients with diabetes. The clinical features and radiological assessments were collected and analyzed. We tracked the dynamic changes in laboratory parameters and clinical outcomes during the hospitalization period. Multivariate analysis was performed using logistic regression to analyze the risk factors that predict the severity of COVID-19 with T2DM. RESULTS: Firstly, compared with the nondiabetic group, the COVID-19 with T2DM group had a higher erythrocyte sedimentation rate (ESR) and levels of C-reactive protein (CRP), interleukin 6 (IL-6), tumor necrosis factor alpha (TNF-α), and procalcitonin (PCT) but lower lymphocyte counts and T lymphocyte subsets, including CD3+ T cells, CD8+ T cells, CD4+ T cells, CD16 + CD56 cells, and CD19+ cells. Secondly, compared with group A, group C had higher levels of Fasting blood glucose (FBG), IL-6, TNF-α, and neutrophils but lower lymphocyte, CD3+ T cell, CD8+ T cell, and CD4+ T cell counts. Similarly, group D had higher FBG, IL-6 and TNF-α levels and lower lymphocyte, CD3+ T cell, CD8+ T cell, and CD4+ T cell counts than group B. Thirdly, binary logistic regression analysis showed that HbA1c, IL-6, and lymphocyte count were risk factors for the severity of COVID-19 with T2DM. Importantly, COVID-19 patients with T2DM were more likely to worsen from moderate to severe COVID-19 than nondiabetic patients. Of note, lymphopenia and inflammatory responses remained more severe throughout hospitalization for COVID-19 patients with T2DM. CONCLUSION: Our data suggested that COVID-19 patients with T2DM are more likely to develop severe COVID-19 than those without T2DM and that hyperglycemia associated with the lymphopenia and inflammatory responses in COVID-19 patients with T2DM.

Dual Raster-Scanning Photoacoustic Small-Animal Imager for Vascular Visualization.

Imaging of vascular networks on small animals has played an important role in basic biomedical research. Photoacoustic imaging technology has great potential for application in the imageology of small animals. The wide-field photoacoustic imaging of small animals can provide images with high spatiotemporal resolution, deep penetration, and multiple contrasts. Also, the real-time photoacoustic imaging system is desirable to observe the hemodynamic activities of small-animal vasculature, which can be used to research the dynamic monitoring of small-animal physiological features. Here, a dual-raster-scanning photoacoustic imager is presented, featuring a switchable double-mode imaging function. The wide-field imaging is driven by a two-dimensional motorized translation stage, while the real-time imaging is realized with galvanometers. By setting different parameters and imaging modes, in vivo visualization of small-animal vascular network can be performed. The real-time imaging can be used to observe pulse change and blood flow change of drug-induced, etc. The wide-field imaging can be used to track the growth change of tumor vasculature. These are easy to be adopted in various areas of basic biomedicine research.

Cadmium sulfate induces apoptosis in planarians.

With rapid socio-economic development, heavy metal pollution in water has become common and affects both environment and human health. Cadmium (Cd) has been recognized as one of the heavy metals which cause acute or chronic toxic effects if ingested. Although its toxicity is undisputed, the underlying molecular mechanisms in vivo are not fully understood. Planarians, a model organism famous for their regenerative prowess, have long been utilized to study the effects of chemical exposure. In this study, we observed apoptosis with TUNEL assay in planarians induced by cadmium sulfate (CdSO4) in a dose-dependent manner. The apoptosis-related genes were detected with quantitative RT-PCR. Significant changes in c-Myc, P53, and BcL-2 were indicated, which may play a partial role in the regulation of the process of apoptosis in the planarians. H&E staining showed that Cd had obvious biological toxicity in the planarians. Here, new insights on metal toxicity mechanisms are provided, contributing to understand how CdSO4 induces the pathological and physiological processes of apoptosis in the living bodies. Meanwhile, planarians are proved to be a freshwater pollution indicator and toxicological research model.

Photoacoustic and ultrasound (PAUS) dermoscope with high sensitivity and penetration depth by using a bimorph transducer.

A bimorph transducer was proposed to improve the detection sensitivity and imaging depth of photoacoustic and ultrasound (PAUS) dermoscope. By applying the bimorph transducer, the imaging depth and sensitivity of PAUS dermoscope were enhanced by simultaneously improving excitation efficiency and reception bandwidth. The integrated design of the imaging head of the dermoscope makes it highly convenient for detecting human skin. The PAUS imaging performance was demonstrated via visualizing subcutaneous tumor and depicting full structures of different skin layers from epidermis to subcutaneous tissue. The results confirm that the dermoscope with the bimorph transducer is well suited for PA and US dual-modality imaging, which can provide multi-information for skin disease.

Wide-field monitoring and real-time local recording of microvascular networks on small animals with a dual-raster-scanned photoacoustic microscope.

Photoacoustic microscopy (PAM) provides a new method for the imaging of small-animals with high-contrast and deep-penetration. However, the established PAM systems have suffered from a limited field-of-view or imaging speed, which are difficult to both monitor wide-field activity of organ and record real-time change of local tissue. Here, we reported a dual-raster-scanned photoacoustic microscope (DRS-PAM) that integrates a two-dimensional motorized translation stage for large field-of-view imaging and a two-axis fast galvanometer scanner for real-time imaging. The DRS-PAM provides a flexible transition from wide-field monitoring the vasculature of organs to real-time imaging of local dynamics. To test the performance of DRS-PAM, clear characterization of angiogenesis and functional detail was illustrated, hemodynamic activities of vasculature in cerebral cortex of a mouse were investigated. Furthermore, response of tumor to treatment were successfully monitored during treatment. The experimental results demonstrate the DRS-PAM holds the great potential for biomedical research of basic biology.

Long non-coding RNA MYOSLID functions as a competing endogenous RNA to regulate MCL-1 expression by sponging miR-29c-3p in gastric cancer.

OBJECTIVE: Long non-coding RNA (lncRNA) has become an important regulator of many human malignancies. However, the biological role and clinical significance of most lncRNA in gastric cancer (GC) remain unclear. METHODS: We investigate the biological function, mechanism of action and clinical expression of lncRNA MYOSLID in GC. First, we analysed the differential expression of lncRNA MYOSLID in GC tissues and non-cancerous tissues by analysing the sequencing data obtained from The Cancer Genome Atlas. Subsequently, we verified that lncRNA MYOSLID regulates the proliferation and apoptosis of GC cells by acting as a ceRNA against miR-29c-3p. The nude mouse xenograft was used to further confirm the functional significance of lncRNA MYOSLID in vivo. RESULTS: We found for the first time that the expression of lncRNA MYOSLID was significantly up-regulated in GC tissues, and the up-regulation of lncRNA MYOSLID in GC was correlated with tumour size, AJCC stage, depth of invasion and survival time. In addition, apoptosis and growth arrest can be induced in vitro after knockdown of lncRNA MYOSLID, which inhibits tumorigenesis in mouse xenografts in vivo. Further in-depth studies revealed that lncRNA MYOSLID acts as a ceRNA of miR-29c-3p, resulting in de-repression of its downstream target gene MCL-1. CONCLUSION: The lncRNA MYOSLID-miR-29c-3p-MCL-1 axis plays a key role in the development of GC. Our findings may provide potential new targets for the diagnosis and treatment of human GC.

Efficacy and safety of bupropion hydrochloride extended-release versus escitalopram oxalate in Chinese patients with major depressive disorder: Results from a randomized, double-blind, non-inferiority trial.

BACKGROUND: This study evaluated the non-inferiority of bupropion extended-release (XL) compared to escitalopram for acute-phase treatment of Chinese patients with major depressive disorder (MDD). METHODS: This randomized (1:1), double-blind, active-control study conducted between February 2015 and October 2016 included patients with MDD (DSM-IV) (N = 538). The treatment phase had three dose levels (level 1 [Week 1], level 2 [Week 2-4], and level 3 [Week 5-8]), which included either bupropion XL 150 mg, 300 mg, 300 mg or escitalopram 10 mg, 10 mg, 10-20 mg (once-daily), respectively. Primary outcome was mean change from baseline in Hamilton Depression Rating Scale-17 (HAMD-17) total score at Week 8. RESULTS: Overall, 534 patients (bupropion XL, n = 266; escitalopram, n = 268) received at least one dose of study medication. The least square mean (standard error) change from baseline in HAMD-17 total score at Week 8 was -14.5 (0.41) in bupropion XL group and -15.4 (0.39) in escitalopram group (mean difference: 0.8 [-0.27, 1.94]). The response rate was 69.6% versus 72.9%, remission rate was 39.7% versus 47.2%, sustained response rate was 51.6% versus 56.3%, and sustained remission rate was 25.5% versus 28.6%, respectively, for bupropion XL versus escitalopram group. Adverse events were reported by 313 patients (bupropion XL, n = 157; escitalopram, n = 156); the most common on-treatment adverse event in both groups was nausea (10.5% versus 18.7%, respectively). LIMITATIONS: A non-inferiority short-term (8 weeks) study without a placebo arm. CONCLUSION: Results from this study demonstrated that the efficacy of bupropion XL was non-inferior to that of escitalopram in Chinese patients with MDD.

Miniaturized photoacoustic probe for in vivo imaging of subcutaneous microvessels within human skin.

BACKGROUND: Subcutaneous microvascular visualization is important for accurate diagnosis and precise treatment of those diseases that are associated with subcutaneous microangiopathy. Pure optical imaging technology and ultrasound imaging technology are commonly used to observe subcutaneous blood vessels non-invasively. However, pure optical imaging is limited to visualizing superficial skin features due to the strong scattering of light by biological tissues, while ultrasound imaging which can detect deep tissues has poor resolution and low contrast to reveal microvascular networks. This results in a lack of intuitive understanding of the disease lesion. METHODS: A miniaturized photoacoustic (PA) probe, which is capable of imaging subcutaneous microvessels with high resolution and deep penetration, was built in this work. The probe is small enough to be hand-held and takes 16 seconds to obtain a maximum amplitude projection image of 400×400 pixels with the imaging area of 2×2 mm2. RESULTS: The miniaturized PA probe was measured to have a lateral resolution of about 8.9 µm and an imaging depth of about 2.4 mm. Besides, in vivo animal experiments and human skin imaging have been implemented. The results show that the miniaturized PA probe not only visualizes the subcutaneous microvessels, but also obtains quantitative information such as the diameters and the depths of blood vessels. CONCLUSIONS: The miniaturized PA probe has potential been used into clinic, and providing quantitative blood vessel information for the diagnosis and monitoring of vascular diseases.

Tenascin-c renders a proangiogenic phenotype in macrophage via annexin II.

Tenascin-c is an extracellular matrix glycoprotein, the expression of which relates to the progression of atherosclerosis, myocardial infarction and heart failure. Annexin II acts as a cell surface receptor of tenascin-c. This study aimed to delineate the role of tenascin-c and annexin II in macrophages presented in atherosclerotic plaque. Animal models with atherosclerotic lesions were established using ApoE-KO mice fed with high-cholesterol diet. The expression of tenascin-c and annexin II in atherosclerotic lesions was determined by qRT-PCR, Western blot and immunohistochemistry analysis. Raw 264.7 macrophages and human primary macrophages were exposed to 5, 10 and 15 µg/ml tenascin-c for 12 hrs. Cell migration as well as the proangiogenic ability of macrophages was examined. Additionally, annexin II expression was delineated in raw 264.7 macrophages under normal condition (20% O2 ) for 12 hrs or hypoxic condition (1% O2 ) for 6-12 hrs. The expression of tenascin-c and annexin II was markedly augmented in lesion aorta. Tenascin-c positively regulated macrophage migration, which was dependent on the expression of annexin II in macrophages. VEGF release from macrophages and endothelial tube induction by macrophage were boosted by tenascin-c and attenuated by annexin II blocking. Furthermore, tenascin-c activated Akt/NF-κB and ERK signalling through annexin II. Lastly, hypoxia conditioning remarkably facilitates annexin II expression in macrophages through hypoxia-inducible factor (HIF)-1α but not HIF-2α. In conclusion, tenascin-c promoted macrophage migration and VEGF expression through annexin II, the expression of which was modulated by HIF-1α.

Ubenimex attenuates acquired sorafenib resistance in renal cell carcinoma by inhibiting Akt signaling in a lipophagy associated mechanism.

Sorafenib is used as first line treatment of renal cell carcinoma (RCC) due to the poor sensitivity to radiotherapy and chemotherapy of this malignancy; however, acquired resistance limits the application of sorafenib and its analogues. In this study, we explored a new strategy to overcome acquired resistance to sorafenib. The RCC cell lines 786-O and ACHN were cultured in presence of increasing concentrations of sorafenib to generate sorafenib-resistant cell lines, 786-O-R and ACHN-R. Interestingly, treatment with ubenimex (0.25 mg/ml) and 3-MA (2 mM) restored the sensitivity of resistant cell lines to sorafenib, indicating the involvement of autophagy in acquired resistance. High levels of autophagy flux were observed in resistant cells, and the opposite effects of ubenimex and 3-MA suggested a complex role for autophagy. While 3-MA abolished protection in sorafenib-resistant cells, ubenimex induced uncontrolled autophagy and autophagic cell death. Lipophagy, characterized by a lipid droplet cargo, was observed in RCC tissues and cells. In sorafenib-resistant cells, ubenimex inhibited the Akt signaling pathway that regulates autophagy. In summary, lipophagy participates in sorafenib-resistance of RCC, which could be reversed by interventions targeting the Akt pathway.