Identification of early events in serrated pathway colorectal tumorigenesis by using digital spatial profiling.

INTRODUCTION: The colorectal serrated pathway involves precursor lesions known as sessile serrated lesions (SSL) and traditional serrated adenomas (TSA). Mutations in BRAF or KRAS are crucial early events in this pathway. Additional genetic and epigenetic changes contribute to the progression of these lesions into high-grade lesions and, eventually, invasive carcinoma. METHODS: We employed digital spatial profiling to investigate the transcriptional changes associated with SSL and TSA. The genes identified are confirmed by immunohistochemical (IHC) staining. Colorectal (CRC) cell lines with CEACAM6 overexpression and knockdown were established to study the roles of CEACAM6 on tumorigenesis of CRC. RESULTS: Ten genes were upregulated in SSL and TSA, and seven were upregulated in both types of lesions. IHC staining confirmed overexpression of CEACAM6, LCN2, KRT19, and lysozyme in SSL and TSA. CEACAM6 expression is an early event in the serrated pathway but a late event in the conventional pathway. Using cell line models, we confirmed that CEACAM6 promotes CRC cells' proliferation, migration, and invasion abilities. CONCLUSION: These results highlight that the transcriptional changes in the early stages of tumorigenesis exhibit relative uniformity. Identifying these early events may hold significant promise in elucidating the mechanisms behind tumor initiation.

Design and synthesis of versatile ligand precursors based on phosphonium ylides for palladalactam formation and catalytic investigation.

A new series of ligand precursors designed for the synthesis of palladalactams has been developed. These precursors are easily accessible through a one-step reaction involving 2-chloro-N-phenylacetamide and a wide choice of various monophosphines, offering tunable electronic and steric properties within the ligand framework. The stability of both ligand precursors and resulting palladalactams in ambient air enhances their practical applicability. A newly synthesized palladalactam, featuring an electron-donating triethylphosphine moiety on the anionic phosphonium ylide ligand scaffold exhibited promising catalytic activities in the Mizoroki-Heck coupling reaction between aryl chlorides and alkenes. Theoretical calculations further affirmed that the ligand system in the complex is the most electron-donating, forming the strongest Pd-C bond compared to other complexes with alternative phosphine moieties.

Laser-induced microbubble as an in vivo valve for optofluidic manipulation in living Mice's microvessels.

Optofluidic regulation of blood microflow in vivo represents a significant method for investigating illnesses linked to abnormal changes in blood circulation. Currently, non-invasive strategies are limited to regulation within capillaries of approximately 10 µm in diameter because the adaption to blood pressure levels in the order of several hundred pascals poses a significant challenge in larger microvessels. In this study, using laser-induced microbubble formation within microvessels of the mouse auricle, we regulate blood microflow in small vessels with diameters in the tens of micrometers. By controlling the laser power, we can control the growth and stability of microbubbles in vivo. This controlled approach enables the achievement of prolonged ischemia and subsequent reperfusion of blood flow, and it can also regulate the microbubbles to function as micro-pumps for reverse blood pumping. Furthermore, by controlling the microbubble, narrow microflow channels can be formed between the microbubbles and microvessels for assessing the apparent viscosity of leukocytes, which is 76.9 ± 11.8 Pa·s in the in vivo blood environment. The proposed design of in vivo microbubble valves opens new avenues for constructing real-time blood regulation and exploring cellular mechanics within living organisms.

Dual targeting total saponins of Pulsatilla of natural polymer crosslinked gel beads with multiple therapeutic effects for ulcerative colitis.

Oral colon targeted drug delivery system (OCTDDS) is desirable for the treatment of ulcerative colitis (UC). In this study, we designed a partially oxidized sodium alginate-chitosan crosslinked microsphere for UC treatment. Dissipative particle dynamics (DPD) was used to study the formation and enzyme response of gel beads from a molecular perspective. The formed gel beads have a narrow particle size distribution, a compact structure, low cytotoxicity and great colon targeting in vitro and in vivo. Animal experiments demonstrated that gel beads promoted colonic epithelial barrier integrity, decreased the level of pro-inflammatory factors, accelerated the recovery of intestinal microbial homeostasis in UC rats and restored the intestinal metabolic disorders. In conclusion, our gel bead is a promising approach for the treatment of UC and significant for the researches on the pathogenesis and treatment mechanism of UC.

STAT3 mediates ECM stiffness-dependent progression in ovarian cancer.

The treatment of ovarian cancer remains a medical challenge and its malignant progression is connected with obvious changes in both tissue and cell stiffness. However, the accurate mechanical-responsive molecules and mechanism remains unclear in ovarian cancer. Based on our previous results combined with the crucial regulatory role of STAT3 in the malignant progression of various cancer types, we want to investigate the relationship between STAT3 and matrix stiffness in ovarian cancer and further explore the potential mechanisms. Collagen-coated polyacrylamide gels (1, 6, and 60 kPa) were prepared to mimic soft or hard matrix stiffness. Western blotting, qRT-PCR, flow cytometry, IHC, EdU assays, and TEM were used to evaluate the effect of STAT3 in vitro under different matrix stiffnesses. Furthermore, a BALB/c nude mouse model was established to assess the relationship in vivo. Our results confirmed the differential expression of STAT3/p-STAT3 not only in normal and malignant ovarian tissues but also under different matrix stiffnesses. Furthermore, we verified that STAT3 was a mechanically responsive gene both in vitro and in vivo, and the mechanical response was carried out by altering the migration-related molecules (TNFAIP1) and adhesion-related molecules (LPXN, CNN3). The novel findings suggest that STAT3, a potential therapeutic target for clinical diagnosis and treatment, is a mechanically responsive gene that responds to matrix stiffness, particularly regulation in migration and adhesion in the progression of ovarian cancer.

Optical assembly of multi-particle arrays by opto-hydrodynamic binding of microparticles close to a one-dimensional chain of magnetic microparticles.

Two-dimensional materials possess a large number of interesting and important properties. Various methods have been developed to assemble two-dimensional aggregates. Assembly of colloidal particles can be achieved with laser-heating-induced thermal convective flow. In this paper, an opto-hydrodynamic binding method is proposed to assemble colloidal particles dispersed in a solution into multilayer structures. First, we use polystyrene (PS) microspheres to study the feasibility and characteristics of the assembly method. PS microspheres and monodispersed magnetic silica microspheres (SLEs) are dispersed in a solution to form a binary mixture system. Under the action of an external uniform magnetic field, SLEs in the solution form chains. An SLE chain is heated by a laser beam. Due to the photothermal effect, the SLE chain is heated to produce a thermal gradient, resulting in thermal convection. The thermal convection drives the PS beads to move toward the heated SLE chain and finally stably assemble into multilayer aggregates on both sides of the SLE chain. The laser power affects the speed and result of the assembly. When the laser power is constant, the degree of constraint of the PS microbeads in different layers is also different. At the same time, this method can also assemble the biological cells, and the spacing of different layers of cells can be changed by changing the electrolyte concentration of the solution. Our work provides an approach to assembling colloidal particles and cells, which has a potential application in the analysis of the collective dynamics of microparticles and microbes.

Optothermal Microparticle Oscillator Induced by Marangoni and Thermal Convection.

The light-fueled microparticle oscillator, exemplifying sustained driving in a static light source, potentially holds applications in fundamental physics, cellular manipulation, fluid dynamics, and various other soft-matter systems. The challenges of photodamage due to laser focusing on particles and the control of the oscillation direction have always been two major issues for microparticle oscillators. Here, we present an optical-thermal method for achieving a 3D microparticle oscillator with a fixed direction by employing laser heating of the gold film surface. First, the microparticle oscillation without direction limitation is studied. The photothermal conversion originates from the laser heating of a gold film. The oscillation mechanism is the coordination of the forces exerted on the particles, including the thermal convective force, thermophoresis force, and gravity. Subsequently, the additional Marangoni convection force, generated by the temperature gradient on the surface of a microbubble, is utilized to control the oscillation direction of the microparticle. Finally, a dual-channel oscillation mode is achieved by utilizing two microbubbles. During the oscillation process, the microparticle is influenced by flow field forces and temperature gradient force, completely avoiding optical damage to the oscillating microparticle.

Development of a rapid aptamer-chemiluminescence sensor for detecting glyphosate pesticide residue in soybeans.

Glyphosate (GLY) is a widely used herbicide worldwide, particularly in cultivating genetically modified soybeans resistant to GLY. However, routine multi-residue analysis does not include GLY due to the complexity of soybean matrix components that can interfere with the analysis. This study presented the development of an aptamer-based chemiluminescence (Apt-CL) sensor for rapidly screening GLY pesticide residue in soybeans. The GLY-binding aptamer (GBA) was developed to bind to GLY specifically, and the remaining unbound aptamers were adsorbed onto gold nanoparticles (AuNPs). The signal was in the form of luminol-H2O2 emission, catalyzed by the aggregation of AuNPs in a chemiluminescent reaction arising from the GLY-GBA complex. The outcomes demonstrated a robust linear relationship between the CL intensity of GLY-GBA and the GLY concentration. In the specificity test of the GBA, only GLY and Profenofos were distinguished among the fifteen tested pesticides. Furthermore, the Apt-CL sensor was conducted to determine GLY residue in organic soybeans immersed in GLY as a real sample, and an optimal linear concentration range for detection after extraction was found to be between 0.001 and 10 mg/L. The Apt-CL sensor exploits the feasibility of real-time pesticide screening in food safety.

Field Measurement of the Dynamic Interaction between Urban Surfaces and Microclimates in Humid Subtropical Climates with Multiple Sensors.

Forcing pathways between urban surfaces (impervious and pervious pavers) and near-surface air temperature were measured and investigated with a network of multiple sensors. Utilizing field data measured between April 2021 and May 2022, and assuming that the influential variables follow the basic heat-transfer energy-balance equations, multiple regression-based statistical models were built to predict the surface temperature and near-surface air temperature (0.05 m, 0.5 m, 1 m, 2 m, and 3 m) of one impervious paver site and one pervious paver site in Taipei City, Taiwan. Evaporative cooling was found to be more influential on the pervious paver with a statistically significant influence on the microclimate up to 1.8 m (and up to 0.7 m for the impervious paver), using in situ data with an ambient air temperature higher than 24 °C. The surface temperature is mainly affected by solar shortwave radiation and ambient air temperature. As for near-surface air temperature, ambient air temperature is the most influential factor, followed by surface temperature. The importance of surface temperature indicates the influence of upwelling longwave radiation on the microclimate. The predictive equations show that pervious surfaces can help cities with hot and humid climates fight the changing climate in the future.

One-Step Synthesis of 3-(Fmoc-amino acid)-3,4-diaminobenzoic Acids.

A one-step method for synthesizing 3-(Fmoc-amino acid)-3,4-diaminobenzoic acids was used to prepare preloaded diaminobenzoate resin. The coupling of free diaminobenzoic acid and Fmoc-amino acids gave pure products in 40-94% yield without any purification step in addition to precipitation except for histidine. For the proline residue, crude products were collected and used for solid-phase peptide synthesis to give a moderate yield of a pentapeptide. In addition, this method was used to prepare unusual amino acid derivatives, namely, (2-naphthyl) alanine and 6-aminohexanoic acid derivatives, in 50 and 65% yield, respectively.

Measurement of red blood cell deformability during morphological changes using rotating-glass-plate-based scanning optical tweezers.

It is important to measure the deformability of red blood cells (RBCs) before transfusion, which is a key factor in the gas transport ability of RBCs and changes during storage of RBCs in vitro. Moreover, the morphology of RBCs also changes during storage. It is proposed that the change in morphology is related to the change in deformability. However, the efficiency of typical methods that use particles as handles is low, especially in the deformability measurement of echinocyte and spherocytes. Therefore, the deformability of RBCs with different morphologies is hard to be measured and compared in the same experiment. In this study, we developed a cost-effective and efficient rotating-glass-plate-based scanning optical tweezers device for the measurement of deformability of RBCs. The performance of this device was evaluated, and the deformability of three types of RBCs was measured using this device. Our results clearly show that the change of erythrocyte morphology from discocyte to echinocyte and spherocyte during storage in vitro is accompanied by a decrease in deformability.

Deformability of mouse erythrocytes in different diluents measured using optical tweezers.

Optical tweezers are widely used to measure the mechanical properties of erythrocytes, which is crucial to the study of pathology and clinical diagnosis of disease. During the measurement, the blood sample is diluted and suspended in an exogenous physiological fluid, which may affect the elastic properties of the cells in vitro. Here, we investigate the effect of different diluents on the elastic properties of mouse erythrocytes by quantitatively evaluating their elastic constants using optical tweezers. The diluents are plasma extracted from mouse blood, veterinary blood diluent (V-52D), Dulbecco's modified Eagle's medium (DMEM), phosphate-buffered saline (PBS), and normal saline (NS). To create an environment that closely resembles in vivo conditions, the experiment is performed at 36.5 °C. The results show that the spring constant of mouse erythrocytes in plasma is 6.23 ± 0.41 µN m-1. The elasticity of mouse erythrocytes in V-52D and DMEM is 8.21 ± 0.91 and 6.95 ± 0.85 µN m-1, which are higher than that in plasma extracted from blood, whereas, the elasticity in PBS and NS is 4.23 ± 0.85 and 4.68 ± 0.79 µN m-1, which are less than that in plasma extracted from blood. At last, we observe the size and circularity of erythrocytes in different diluents, and consider that the erythrocyte diameter and circularity may affect cell deformability. Our results provide a reference of the diluent choice for measuring the mechanical properties of erythrocytes in vitro.

Optical trapping of microparticles with two tilted-focused laser beams.

We present an optical method for the manipulation of microparticles using two tilted-focused beams. First, the action on the microparticles is studied with a single tilted-focused beam. The beam is used to drive the directional motion of a dielectric particle. When the optical scattering force is larger than the optical gradient force, the particle is pushed to the tilted side of the optical axis by the optical force. Second, two tilted-focused beams with the same power and complementary tilt angles are used to assemble an optical trap. The trap can be used to realize the optical trapping of the dielectric particles and opto-thermal trapping of the light absorbing particles. The trapping mechanism is the balance of the forces exerted on the particles, including the optical scattering force, optical gradient force, gravity, and thermal gradient force. The trap center is away from the focal spots, which effectively prevents the laser beam from being focused on the trapped object.

An opto-thermal approach for assembling yeast cells by laser heating of a trapped light absorbing particle.

Cell assembly has important applications in biomedical research, which can be achieved with laser-heating induced thermal convective flow. In this paper, an opto-thermal approach is developed to assemble the yeast cells dispersed in solution. At first, polystyrene (PS) microbeads are used instead of cells to explore the method of microparticle assembly. The PS microbeads and light absorbing particles (APs) are dispersed in solution and form a binary mixture system. Optical tweezers are used to trap an AP at the substrate glass of the sample cell. Due to the optothermal effect, the trapped AP is heated and a thermal gradient is generated, which induces a thermal convective flow. The convective flow drives the microbeads moving toward and assembling around the trapped AP. Then, the method is used to assemble the yeast cells. The results show that the initial concentration ratio of yeast cells to APs affects the eventual assembly pattern. The binary microparticles with different initial concentration ratios assemble into aggregates with different area ratios. The experiment and simulation results show that the dominant factor in the area ratio of yeast cells in the binary aggregate is the velocity ratio of the yeast cells to the APs. Our work provides an approach to assemble the cells, which has a potential application in the analysis of microbes.

Synergistic anticancer effect of flavonoids from Sophora alopecuroides with Sorafenib against hepatocellular carcinoma.

Sorafenib (SF), a multi-kinase inhibitor, is the first FDA-approved systemic chemotherapy drug for advanced hepatocellular carcinoma (HCC). However, its clinical application is limited by severe toxicity and side effects associated with high applied doses. Sophora alopecuroides L. is traditionally used as Chinese herbal medicine for treating gastrointestinal diseases, bacillary dysentery, viral hepatitis, and other diseases, and exerts an important role in anti-tumor. Hence, we investigated the synergistic actions of seventeen flavonoids from this herb combined with SF against HCC cell lines and their primary mechanism. In the experiment, most compounds were found to prominently enhance the inhibitory effects of SF on HCC cells than their alone treatment. Among them, three compounds leachianone A (1), sophoraflavanone G (3), and trifolirhizin (17) exhibited significantly synergistic anticancer activities against MHCC97H cells at low concentration with IC50 of SF reduced by 5.8-fold, 3.6-fold, and 3.5-fold corresponding their CI values of 0.49, 0.66, and 0.46 respectively. Importantly, compounds 3 or 17 combined with SF could synergistically induce MHCC97H cells apoptosis via the endogenously mitochondrial-mediated apoptotic pathway, involving higher Bax/Bcl-2 expressions with the activation of caspase-9 and -3, and arrest the cell cycle in G1 phases. Strikingly, this synergistic effect was also closely related to the co-suppression of ERK and AKT signaling pathways. Furthermore, compound 3 significantly enhanced the suppression of SF on tumor growth in the HepG2 xenograft model, with a 79.3% inhibition ratio at high concentration, without systemic toxicity, compared to either agent alone. These results demonstrate that the combination treatment of flavonoid 3 and SF at low doses exert synergistic anticancer effects on HCC cells in vitro and in vivo.

Micropeptides: origins, identification, and potential role in metabolism-related diseases / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

With the development of modern sequencing techniques and bioinformatics, genomes that were once thought to be noncoding have been found to encode abundant functional micropeptides (miPs), a kind of small polypeptides. Although miPs are difficult to analyze and identify, a number of studies have begun to focus on them. More and more miPs have been revealed as essential for energy metabolism homeostasis, immune regulation, and tumor growth and development. Many reports have shown that miPs are especially essential for regulating glucose and lipid metabolism and regulating mitochondrial function. MiPs are also involved in the progression of related diseases. This paper reviews the sources and identification of miPs, as well as the functional significance of miPs for metabolism-related diseases, with the aim of revealing their potential clinical applications.

Advances in mechanism and regulatory factors of corneal stromal repair / 国际眼科杂志(Guoji Yanke Zazhi)

Corneal stromal is an important structure to maintain corneal transparency. The corneal stroma can be injured by trauma, infection and surgery. Therefore, corneal stromal wound starts repair with phenotype changes in stromal cell, extracellular matrix remodeling and immune cells migration. The corneal scar was the leading cause of blindness worldwide, which can be caused by corneal stromal fibrosis from increased myofibroblasts and deposited extracellular matrix after sever damage. At present, corneal transplantation is the main treatment for corneal scar, which has limited therapeutic effect because of corneal donor shortage, surgical requirements and the risk of postoperative immune rejection. Recently, great progress has been made in the study of control mechanism of corneal stromal wound healing with various molecules, cells and tissues. This paper reviews the repair mechanism of corneal stromal injury and the regulation mechanism of cause of corneal injury, corneal structure and molecule factors towards corneal stromal injury. It aims at providing new ideas for exploring the mechanism of corneal stromal repair and regeneration, which is supposed to help prevent corneal scar clinically.

Automatic evaluation system for ultrasound biomicroscopy images of anterior chamber angle based on deep learning algorithm / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To explore the clinical application value of analysis system for ultrasound biomicroscopy(UBM)images of anterior chamber angle(ACA)based on deep learning algorithm.METHODS: A total of 4 196 UBM images were obtained from 675 patients(1 130 eyes)at the Eye Center of Renmin Hospital of Wuhan University from January 2021 to June 2022 were collected to build an image dataset. Using Unet++network to automatically segment ACA tissue, a support vector machine(SVM)algorithm was developed to automatically classify opening and closing of chamber angle, and an algorithm to automatically locate the sclera spur and measure ACA parameters was developed. Furthermore, a total of 631 UBM images of 127 subjects(221 eyes)at Huangshi Aier Eye Hospital and 594 UBM images of 188 subjects(257 eyes)at Zhongnan Hospital of Wuhan University were selected to evaluate the performance of the system under different environments.RESULTS: The accuracy of the analysis system constructed in this study for chamber angle opening and closing was 95.71%. The intra-class correlation coefficient(ICC)values of all ACA angle parameters were greater than 0.960. ICC values of all ACA thickness parameters were greater than 0.884. The accurate measurement of ACA parameters depended in part on the accurate location of the scleral spur.CONCLUSION: The intelligent analysis system constructed in this study can accurately and effectively evaluate ACA images automatically and is a potential screening tool for the rapid identification of ACA structures.

An opto-thermal approach for rotating a trapped core-shell magnetic microparticle with patchy shell.

The ability to trap and rotate magnetic particles has important applications in biophysical research and optical micromachines. However, it is difficult to achieve the spin rotation of magnetic particles with optical tweezers due to the limit in transferring spin angular momentum of light. Here, we propose a method to obtain controlled spin rotation of a magnetic microparticle by the phoretic torque, which is originated from inhomogeneous heating of the microparticle's surface. The microparticle is trapped and rotated nearby the laser focus center. The rotation frequency is several Hertz and can be controlled by adjusting the laser power. Our work provides a method to the study of optical rotation of microscopic magnetic particles, which will push toward both translational and rotational manipulation of the microparticles simultaneously in a single optical trap.

Non-Invasive Dynamic Reperfusion of Microvessels In Vivo Controlled by Optical Tweezers.

Distributive shock is considered to be a condition of microvascular hypoperfusion, which can be fatal in severe cases. However, traditional therapeutic methods to restore the macro blood flow are difficult to accurately control the blood perfusion of microvessels, and the currently developed manipulation techniques are inevitably incompatible with biological systems. In our approach, infrared optical tweezers are used to dynamically control the microvascular reperfusion within subdermal capillaries in the pinna of mice. Furthermore, we estimate the effect of different optical trap positions on reperfusion at branch and investigate the effect of the laser power on reperfusion. The results demonstrate the ability of optical tweezers to control microvascular reperfusion. This strategy allows near-noninvasive reperfusion of the microvascular hypoperfusion in vivo. Hence, our work is expected to provide unprecedented insights into the treatment of distributive shock.