An ULK1/2-PXN mechanotransduction pathway suppresses breast cancer cell migration.

The remodeling and stiffening of the extracellular matrix (ECM) is a well-recognized modulator of breast cancer progression. How changes in the mechanical properties of the ECM are converted into biochemical signals that direct tumor cell migration and metastasis remain poorly characterized. Here, we describe a new role for the autophagy-inducing serine/threonine kinases ULK1 and ULK2 in mechanotransduction. We show that ULK1/2 activity inhibits the assembly of actin stress fibers and focal adhesions (FAs) and as a consequence impedes cell contraction and migration, independent of its role in autophagy. Mechanistically, we identify PXN/paxillin, a key component of the mechanotransducing machinery, as a direct binding partner and substrate of ULK1/2. ULK-mediated phosphorylation of PXN at S32 and S119 weakens homotypic interactions and liquid-liquid phase separation of PXN, impairing FA assembly, which in turn alters the mechanical properties of breast cancer cells and their response to mechanical stimuli. ULK1/2 and the well-characterized PXN regulator, FAK/Src, have opposing functions on mechanotransduction and compete for phosphorylation of adjacent serine and tyrosine residues. Taken together, our study reveals ULK1/2 as important regulator of PXN-dependent mechanotransduction.

Impact of different dietary fibre sources on production performance, bacterial composition and metabolites in the caecal contents of rabbits.

This study was conducted to investigate the effects of different dietary fibre sources on growth performance, gastrointestinal tract development, caecal fermentation and bacterial composition in the caecal contents of rabbits. A total of 120 35-day-old weaned Minxinan black rabbits were divided into three groups and fed a diet composed of peanut straw powder (Group A), alfalfa powder (Group B) and soybean straw powder (Group C) as the main fibre source. The final body weight and average daily gain in Group B were higher than those in Group C, and the average daily feed intake and feed conversion ratio in Group A were lower than those in Group C (p < 0.05). The relative weights of the stomach, small intestine and caecum of rabbits in Group C were higher than those in Groups of B and A, and the relative weights of the caecal contents in Group C were lower than those in Groups A or B (p < 0.05). The pH value and propionic acid, butyric acid and valeric acid concentrations in the caecum of Group C were lower than those in the caecum of Groups A or B, and the concentration of acetic acid in the caecum was lower (p < 0.05). The dominant microbes in the caecal contents of Minxinan black rabbits were Firmicutes, Bacteroidetes and Proteobacteria at the phylum level, and the number of species, Chao1 index and ACE index measured was different between the B-C and A-C groups (p < 0.05). Different dietary fibre sources could affect the growth performance, gastrointestinal tract development and intestinal microbiota of rabbits, and the nutritional value of alfalfa powder was better than that of peanut straw and soybean straw.

Efficacy and safety of a single intra-articular injection of 6 ml Hylan G-F 20 compared to placebo in Chinese patients with symptomatic knee osteoarthritis : C-SOUND study, a 26-week multicenter double-blind randomized placebo-controlled trial in China.

BACKGROUND: Single 6 ml Hylan G-F 20 injection, is indicated for knee osteoarthritis patients who have failed to respond to non-pharmacologic therapy and/or simple analgesics. To obtain more thorough understanding of the clinical efficacy and safety, a randomized clinical trial was conducted comparing intra-articular (IA) administration of single 6 ml Hylan G-F 20 injection versus placebo in knee OA patients of Chinese ethnicity. METHODS: This was a randomized, multi-center, double-blind, placebo-controlled clinical trial conducted in 21 centers across China. Four hundred forty adults with knee OA received a single 6 ml Hylan G-F 20 or placebo injection and were evaluated for clinical efficacy and safety outcomes over 26 weeks. Western Ontario and McMaster Universities OA (WOMAC) A1 index, treatment-emergent adverse events (TEAEs) and standard safety parameters were measured at pre-injection, and at weeks 1, 4, 8, 12, 16, 20 and 26 post-injection. RESULTS: Four hundred forty patients (male: 98 [22.3%]; female: 342 [77.7%]) were randomized. The mean age [standard deviation (SD)] was 61.5 (7.9) years. All patients were of East Asian ethnicity. Mean WOMAC A1 score at baseline was 5.3 (1.2) and 5.2 (1.3) in single 6 ml Hylan G-F 20 injection and placebo groups, respectively. Significant reductions of WOMAC A1 score were observed in both treatment groups when compared to baseline at 26 weeks post-injection, with the mean reduction of [standard error (SE)/percentage] -2.146 (0.108)/- 40.5% and - 2.271 (0.110) /- 43.7% in the single 6 ml Hylan G-F 20 injection and the placebo groups, respectively. Additionally, clinically important reductions in pain at 26 weeks was reported in 67.0 and 68.2% in single 6 ml Hylan G-F 20 injection and placebo groups (p = 0.36). Regarding safety, TEAEs were similar between the two treatment groups (hylan G-F 20 single: 61.5%; placebo: 64.5%). CONCLUSIONS: While the magnitude of the effect of a single 6 ml Hylan G-F 20 injection in this study is consistent with previously published literature with respect to the efficacy and safety of the drug, the current study shows a strong IA placebo effect and did not established superiority of single 6 ml Hylan G-F 20 injection over IA placebo in Chinese knee OA patients. TRIAL REGISTRATION: Prospectively registered Jun 16, 2017 at www.clinicaltrials.gov ( NCT03190369 ).

DNA methylation and histone acetylation are involved in Wnt10b expression during the secondary hair follicle cycle in Angora rabbits.

Secondary hair follicles (SHFs) in the Angora rabbit exhibit classic cyclic hair development, but the multiple molecular signals involved in hair cycling are yet to be explored in detail. In the present study, we investigated the expression pattern, methylation and histone H3 acetylation status of Wnt10b, as a molecular signal participating in hair cycling, during the SHF cycle in the Angora rabbit. Expression of Wnt10b at the anagen phase was significantly higher than that at both the telogen and catagen phases, suggesting that Wnt10b might serve as a critical activator during cyclic transition of SHFs. Methylation frequency of the fifth CpG site (CpG5-175 bp) in CpG islands at the anagen phase was lower than that at both the catagen and telogen phases. The methylation status of the CpG5 site was negatively correlated with Wnt10b expression. This indicated that the methylation of CpG5 might participate in Wnt10b transcriptional suppression in SHFs. Furthermore, histone H3 acetylation status in the regions-256~-11 bp and 98 ~ 361 bp were significantly lower at both the catagen and telogen phases than at the anagen phase. The histone H3 acetylation level was significantly positively correlated with Wnt10b expression. This confirmed that histone acetylation was likely involved in upregulating Wnt10b transcription in SHFs. Additionally, potential binding to the transcription factors ZF57 and HDBP was predicted within the CpG5 site. In conclusion, our findings reveal the epigenetic mechanism of Wnt10b transcription and provide a new insight into epigenetic regulation during the SHF cycle in the Angora rabbit.

The functions of ocu-miR-205 in regulating hair follicle development in Rex rabbits.

BACKGROUND: Hair follicles are an appendage of the vertebrate epithelium in the skin that arise from the embryonic ectoderm and regenerate cyclically during adulthood. Dermal papilla cells (DPCs) are the key dermal component of the hair follicle that directly regulate hair follicle development, growth and regeneration. According to recent studies, miRNAs play an important role in regulating hair follicle morphogenesis and the proliferation, differentiation and apoptosis of hair follicle stem cells. RESULTS: The miRNA expression profile of the DPCs from Rex rabbits with different hair densities revealed 240 differentially expressed miRNAs (|log2(HD/LD)| > 1.00 and Q-value≤0.001). Among them, ocu-miR-205-5p was expressed at higher levels in DPCs from rabbits with low hair densities (LD) than in rabbits with high hair densities (HD), and it was expressed at high levels in the skin tissue from Rex rabbits (P < 0.05). Notably, ocu-miR-205 increased cell proliferation and the cell apoptosis rate, altered the progression of the cell cycle (P < 0.05), and modulated the expression of genes involved in the PI3K/Akt, Wnt, Notch and BMP signalling pathways in DPCs and skin tissue from Rex rabbits. It also inhibited the phosphorylation of the CTNNB1 and GSK-3ß proteins, decreased the level of the noggin (NOG) protein, and increased the level of phosphorylated Akt (P < 0.05). A significant change in the primary follicle density was not observed (P > 0.05), but the secondary follicle density and total follicle density (P < 0.05) were altered upon interference with ocu-miR-205-5p expression, and the secondary/primary ratio (S/P) in the ocu-miR-205-5p interfered expression group increased 14 days after the injection (P < 0.05). CONCLUSIONS: In the present study, ocu-miR-205 promoted the apoptosis of DPCs, altered the expression of genes and proteins involved in the PI3K/Akt, Wnt, Notch and BMP signalling pathways in DPCs and skin from Rex rabbits, promoted the transition of hair follicles from the growth phase to the regression and resting phase, and altered the hair density of Rex rabbits.

Prognostic factors for overall survival in patients with spinal metastasis secondary to prostate cancer: a systematic review and meta-analysis.

BACKGROUND: To guide the selection of treatments for spinal metastases, the expected survival time is one of the most important determinants. Few scoring systems are fully applicable for spinal metastasis secondary to prostate cancer (PCa). This study aimed to identify the independent factors to predict the overall survival (OS) of patients with spinal metastases from PCa. METHODS: The PubMed, Embase and CENTRAL were retrieved by two reviewers independently, to identify studies analyzed the prognostic effect of different factors in spinal metastasis from PCa. A systematic review and quantitative meta-analysis was conducted with hazard ratio (HR) and 95% confidence interval (95%CI) as the effect size. RESULTS: A total of 12 retrospective cohort studies (1566 patients) were eligible for qualitative synthesis and 10 for quantitative meta-analyses. The OS was significantly influenced by performance status, visceral metastasis, ambulatory status and time from PCa diagnosis in more than half of the available studies. The meta-analyses demonstrated that OS was significantly influenced by visceral metastasis (HR = 2.24, 95%CI:1.53-3.27, p < 0.001), pre-treatment ambulatory status (HR = 2.64, 95%CI:1.82-3.83, p < 0.001), KPS (HR = 4.45, 95%CI:2.01-9.85, p < 0.001), ECOG (HR = 2.96, 95%CI:2.02-4.35, p < 0.001), extraspinal bone metastasis (HR = 2.04, 95%CI:1.13-3.68, p = 0.018), time developing motor deficit (HR = 1.57, 95%CI:1.30-1.88, p < 0.001) and time from PCa diagnosis (HR = 1.37, 95%CI:1.17-1.59, p < 0.001). CONCLUSIONS: Visceral metastasis, ambulatory status, extraspinal bone metastasis, performance status, time developing motor deficit and time interval from primary tumor diagnosis were significantly associated with the OS for spinal metastasis from PCa. When selecting the treatment modality, clinicians should fully consider the patients' systematic status based on all potential prognostic factors. LEVEL OF EVIDENCE: I Meta-analysis.

Extracorporeal shock wave therapy versus other therapeutic methods for chronic plantar fasciitis.

BACKGROUND: To conduct a meta-analysis comparing the efficacy of general ESWT with that of other therapies and to assess its effectiveness in chronic plantar fasciitis. METHODS: A literature search was performed in PubMed, Embase, Web of Science and the Cochrane Library for information from the earliest date of data collection to March 2018. Studies comparing the benefits and risks of extracorporeal shock wave therapy with those of other therapies for chronic plantar fasciitis were included. Statistical heterogeneity was quantitatively evaluated by a X2 test with the significance set as P<0.10 or I2>50%. RESULTS: Thirteen trials consisting of 1,185 patients were included (637 patients were treated with ESWT; 548 patients, with OT). The results showed that patients treated with ESWT had increased success or improvement rates, an increased modified Roles & Maudsley (RM) score, a reduction of pain scales, reduced return to work time, and fewer complications than patients treated with other therapy methods (P<0.1). CONCLUSIONS: Compared with patients who received other therapies for chronic plantar fasciitis, patients treated with ESWT responded better, had less complications and showed a clear difference in efficacy between ESWT and other therapy in chronic plantar fasciitis. LEVEL OF EVIDENCE: Level IV, therapeutic study.

Effects of Wnt10b on dermal papilla cells via the canonical Wnt/ß-catenin signalling pathway in the Angora rabbit.

Wnt10b is a member of Wnt family that plays a variety of roles in biological functions, including those in the development of hair follicles. To investigate the effect of Wnt10b on hair growth in the Angora rabbit and to determine the underlying molecular mechanism, we cultured dermal papilla (DP) cells with exogenous Wnt10b in vitro. We observed the expressions of downstream critical gene ß-catenin and lymphoid enhancer-binding factor 1 (LEF1) in Wnt/ß-catenin pathway. The levels of ß-catenin mRNA and protein were higher in the Wnt10b group of DP cells than in the Control group, and the mRNA level of LEF1 in the Wnt10b group was higher than in the Control group. Moreover, translocation of ß-catenin from cytoplasm to nucleus was activated in the Wnt10b group. Furthermore, the mRNA levels of the hair follicle-regulatory genes, insulin-like growth factor-1 (IGF-1) and alkaline phosphatase (ALP), and the protein activity of ALP was also upregulated in the Wnt10b group compared to their corresponding levels in the Control group. These data suggest that Wnt10b could activate the canonical Wnt/ß-catenin signalling pathway to induce DP cells in the Angora rabbit. In addition, the proliferation of DP cells was significantly promoted when cultured with Wnt10b for 48 and 72 hr, suggesting that Wnt10b plays a pivotal role in the proliferation and maintenance of DP cells in vitro. In conclusion, this study demonstrates that Wnt10b may promote hair follicle growth in Angora rabbit through the canonical Wnt/ß-catenin signalling pathway that promotes the proliferation of DP cells.

The effects of pulsed electromagnetic fields combined with a static magnetic intramedullary implant on the repair of bone defects: A preliminary study.

There is a basic consensus on the biological effects of pulsed electromagnetic fields (PEMFs) on bone formation and bone reconstruction. PEMFs have been widely used in clinical treatment of osteoporosis, bone nonunion and delayed fracture healing. PEMFs is an intervention method of physiotherapy in vitro. In order to optimize the effect of PEMFs intervention, this study combined with the orthopedics clinic to construct a static magnetic intramedullary implant using NdFeB magnets as components. At the same time, it combines external-pulsed electromagnetic field to achieve locally targeted magnetic microenvironment. Rabbits were randomly divided into a combined magnetic field group (Implantation of static magnetic intramedullary implant in vivo combined with external-pulsed electromagnetic field), pulsed electromagnetic field group and control group. Micro CT and histopathology were used to estimate the effect of each group on bone formation and reconstruction in the early stage (5 weeks) of bone defect repair. Our data showed that the combined magnetic field group had relatively better new bone volume and trabecular structure in the bone defect area. The results showed that the combined magnetic field intervention method was feasible and had relatively preferably osteogenesis promoting effect. This study provides a new idea of magnetic field intervention, and also preliminarily verifies the feasibility of adding magnetic field to traditional orthopedic implant materials. However, the magnetic field strength of implanted materials still needs to be further refined.

Underlying Signaling Pathways and Therapeutic Applications of Pulsed Electromagnetic Fields in Bone Repair.

Pulsed electromagnetic field (PEMF) stimulation, as a prospective, noninvasive, and safe physical therapy strategy to accelerate bone repair has received tremendous attention in recent decades. Physical PEMF stimulation initiates the signaling cascades, which effectively promote osteogenesis and angiogenesis in an orchestrated spatiotemporal manner and ultimately enhance the self-repair capability of bone tissues. Considerable research progresses have been made in exploring the underlying cellular and subcellular mechanisms of PEMF promotion effect in bone repair. Moreover, the promotion effect has shown strikingly positive benefits in the treatment of various skeletal diseases. However, many preclinical and clinical efficacy evaluation studies are still needed to make PEMFs more effective and extensive in clinical application. In this review, we briefly introduce the basic knowledge of PEMFs on bone repair, systematically elaborate several key signaling pathways involved in PEMFs-induced bone repair, and then discuss the therapeutic applications of PEMFs alone or in combination with other available therapies in bone repair, and evaluate the treatment effect by analyzing and summarizing recent literature.

Discrete fringe phase unwrapping algorithm based on Kalman motion estimation for high-speed I/Q-interferometry.

A discrete fringe phase unwrapping algorithm (DFPUA) based on Kalman motion estimation is proposed to accurately demodulate the phases of I/Q-interferometers with deeply under-sampled quadrature signals, thus to break through the limitations of the Nyquist frequency for high-speed measurement. The basic concept of DFPUA is to estimate the current displacement according to the former motion state, then confirm the actual phase integer number by comparing the estimated phase decimal with the actual phase decimal; in this way, peak acceleration/jerk instead of peak velocity becomes the factor that determines the sampling rate. Two types of DFPUA including velocity estimation and velocity-acceleration estimation are illustrated in detail. Simulation experiment results indicate that the DFPUA realizes a significant reduction in the sampling rate and the amount of data for low frequency vibration measurement, proposing a practical approach for high-speed and long-time measurement such as ultra-low frequency vibration calibration.

Association of facet tropism and orientation with lumbar disc herniation in young patients.

Facet tropism and orientation are thought to be associated with lumbar disc herniation (LDH), but the relationship is not well established. Moreover, the effect of facet joint on LDH has not been outlined in young patients. The objective of this study was to investigate the associations of facet joint tropism and orientation with LDH in young patients (18-35 years) by computed tomography (CT). Fifty-three patients with LDH and 129 with neither LDH nor low back pain (18-35 years) were included in this study. The facet joint angles were measured for each facet joint by CT as per the method described by Noren et al. We defined facet tropism as a bilateral angle difference > 5°. Young cases with neither LDH nor low back pain were used as the control group. The results showed that LDH was significantly associated with more coronal facet joint orientation at L1-2 (p = 0.009), L2-3 (p = 0.004), and L3-4 (p = 0.004). No association was established between facet tropism and LDH. This study revealed that facet joint orientation was associated with LDH in young patients (18-35 years); they were more of coronal facing at upper levels. Also, the facet tropism was not associated with LDH.

Lanthanoid tagging via an unnatural amino acid for protein structure characterization.

Lanthanoid pseudo-contact shift (PCS) provides long-range structural information between a paramagnetic tag and protein nuclei. However, for proteins with native cysteines, site-specific attachment may only utilize functional groups orthogonal to sulfhydryl chemistry. Here we report two lanthanoid probes, DTTA-C3-yne and DTTA-C4-yne, which can be conjugated to an unnatural amino acid pAzF in the target protein via azide-alkyne cycloaddition. Demonstrated with ubiquitin and cysteine-containing enzyme EIIB, we show that large PCSs of distinct profiles can be generated for each tag/lanthanoid combination. The DTTA-based lanthanoid tags are associated with large magnetic susceptibility tensors owing to the rigidity of the tags. In particular, introduction of the DTTA-C3 tag affords intermolecular PCSs and enables structural characterization of a transient protein complex between ubiquitin and a UBA domain. Together, we have expanded the repertoire of paramagnetic tags and the applicability of paramagnetic NMR.

Comparison of three-dimensional printing and vacuum freeze-dried techniques for fabricating composite scaffolds.

In this study, the performances of different preparation methods of the scaffolds were analyzed for chondrocyte tissue engineering. Silk fibroin/collagen (SF/C) was fabricated using a vacuum freeze-dried technique and by 3D printing. The porosity, water absorption expansion rates, mechanical properties, and pore sizes of the resulting materials were evaluated. The proliferation and metabolism of the cells was detected at different time points using an MTT assay. Cell morphologies and distributions were observed by histological analysis and scanning electron microscopy (SEM). The porosity, water absorption expansion rate, and Young's modulus of the material obtained via 3D printing were significantly higher than those obtained by the freeze-dried method, while the pore size did not differ significantly between the two methods. MTT assay results showed that the metabolism of cells seeded on the 3D printed scaffolds was more viable than the metabolism on the freeze-dried material. H&E staining of the scaffolds revealed that the number of cells in the 3D printed scaffold was higher in comparison to a similar measurement on the freeze-dried material. Consequently, stem cells grew well inside the 3D printed scaffolds, as measured by SEM, while the internal structure of the freeze-dried scaffold was disordered. Compared with the freeze-dried technique, the 3D printed scaffold exhibited better overall performance and was more suitable for cartilage tissue engineering.

Comparisons of Emu Necrotic Femoral Head Micro Structure Repaired in Two Different Methods.

OBJECTIVE: To compare emu necrotic femoral head micro structure repaired in two different methods. METHODS: Fifteen adult emus were divided into 3 groups (all n=5), and the right femoral head was selected to research. The first group was the control group; in the second group, femoral head necrosis was made by cryogen with liquid nitrogen; and in the third group, femoral head necrosis was made by local pure ethanol injection. Right femurs were taken for micro CT examination,then femoral head micro structures were compared among these three groups. RESULTS: No infection or unexpected death was found in all groups. Compared with normal group, necrotic femoral heads in cryogen group showed that bone mineral density significantly reduced after repaire (P=0.015), trabecular space significantly reduced (P=0.001), bone volume fraction significantly enlarged (P=0.036), bone surface/volume fraction (P=0.032) and trabecular numbers (P=0.002) significantly enlarged; trabecular thickness showed no significant difference (P=0.060). Compared with control group, necrotic femoral heads in ethanol group showed that bone mineral density significantly enlarged after repaire (P=0.001), trabecular thickness (P=0.003) and bone surface/volume fraction (P=0.022) significantly enlarged, trabecular space (P=0.001) and bone volume fraction (P=0.001) significantly reduced; the trabecular numbers showed no significant difference (P=0.143). Compared with ethanol group, necrotic femoral heads in cryogen group showed significant lower bone mineral density after repair (P=0.001), significantly lower bone volume fraction (P=0.001), significantly lower trabecular thickness (P=0.001), significantly higher bone surface/volume fraction (P=0.022) and higher trabecular numbers (P=0.003); the trabecular space showed no significant difference (P=0.398). CONCLUSION: Different repair methods make reconstructed femoral head weight bearing area have different bone structure and bone mineral density, along with different bone trabecular quality.

[Effect of stem fixing methods on stress and bone mineral density in the proximal femur following hip resurfacing arthroplasty].

OBJECTIVE: To construct different 3-D finite element resurfacing arthroplasty of hip models and to explore the biomechanics influences of cement mantle varying in thickness on stress distribution and stressshielding in the proximal femur following hip resurfacing arthroplasty (RSAH), and evaluate the changes of bone mineral density in femur after RSAH , in order to find the adaptive methods to stem fixing. METHODS: A comparative study was carrie out in patients who underwent RSAH. The bone mineral Density of the proximal femur was measured through dual energy X-ray absorptiomenty of the four zone at post-operation and unoperation 6 months. Changes of bone mineral density ratio in proximal femur between post-operation and unoperation were compared and analyzed.the finite element models of RSAH with different thickness of cement mantle and normal femur were developed and stress analysis was performed under the load of body weight. RESULTS: The mean ration of bone mineral density of proximal femur in zone 1, 2 , 3 increased which was no significantly different from other areas (P > 0.05), the mean ration of bone mineral density of proximal femur in zone 4 and 5 increased which was significantly different from other areas (P < 0.05). Comparing with the 0 mm cement mantle model, the ratio of stress shielding of the cancellous bone around the stem in 1 mm cement mantle model has slightly increased, but also was less than 17%. The ratio of stress shielding was increased obviously in the 2 and 3 mm cement mantle mode and more than 17% in 1, 2 and 5 zone. CONCLUSION: The cement around stem not thicker than 1 mm produced slightly effect to the stress decreasing and did not result in marked stress shielding in the proximal femur following RSAH, and the bone stock of proximal femur can be well perserves and recovered.

[Study of animal model of osteonecrosis induced by local ethanol injection in emu].

OBJECTIVE: To establish a new animal model of osteonecrosis of the femoral head by local ethanol injection in emu. METHODS: Eight milliliter ethanol was injected slowly to the operated femoral head with customized probe in twenty adult male emus. Postoperatively, hip magnetic resonance imaging was performed at 1, 4, 8, 12 weeks. After emus were sacrificed, the femurs were collected for micro-computed tomography and histological analysis. RESULTS: No emu demonstrated signs of infection or died unexpectedly. Magnetic resonance imaging examination showed broad edema at proximal femur at 1(th) week, and the edema decreased with time, till local edema at femoral head at the 12(th) week. Histological images showed human-like osteonecrotic changes with active bone repair. There were significant differences in trabecular structure and bone mineral density between the operated and intact femoral heads. No collapse was found 6 months after the operation. CONCLUSIONS: This emu model of femoral head osteonecrosis by local ethanol injection can progress to early stage osteonecrosis. The different repair methods may have certain correlation with the results of osteonecrosis of the femoral heads.

[A method based on endogenous fluorescence determination for screening NAMPT inhibitors].

OBJECTIVE: To establish a method for screening nicotinamide phosphoribosyl transferase (NAMPT) inhibitors based on endogenous fluorescence determination. METHODS: The double mutants of NAMPT, G355C/D393C, was cross-linked by using 1, 4-Bismaleimidobutane (BMB) to block the entrance of enzymatic active site of NAMPT. The binding of compounds to NAMPT was evaluated according to the change of spontaneous fluorescence of NAMPT and BMB-NAMPT with 280 nm excitation and 333 nm emmision. The in vitro enzamatic activity of NAMPT was determined by nuclear magnetic resonance. The cell viability was determined by MTT assay. RESULTS: FK866 significantly decreased the spontaneous fluorescence of NAMPT but not of BMB-NAMPT. Rosmaric, cynarine and 1, 3-dicaffeoylquinic acid also decreased the spontaneous fluorescence of both NAMPT and BMB-NAMPT. However, the inhibition on two proteins was equivalent. FK866 significantly inhibit the catalysis of NAMPT. Rosmarinic acid, cynarine and 1, 3-dicaffeoylquinic acid failed to inhibit the catalysis of NAMPT. FK866 inhibited the viability of A549 cells, but rosmarinic acid, cynarine and 1, 3-dicaffeoylquinic acid did not. CONCLUSION: Endogenous fluorescence spectrometry based on NAMPT and BMB-NAMPT protein can be used for screening compounds that bind with NAMPT, and distinguishing the binding site - either within the enzymatic active site or not. Rosmarinic acid, cynarine and 1, 3-dicoffeoylquinic acid can bind to NAMPT out its enzymatic active site.

Detection and analysis of potential landslides based on SBAS-InSAR technology in alpine canyon region.

The Lancang River flows through the alpine canyon region of southwest China, an area that has experienced frequent geological disasters over the years. Early monitoring of geological hazards is essential for disaster prevention and mitigation. However, traditional ground monitoring techniques are limited by the complex terrain conditions in high-altitude valley regions. In contrast, interferometric synthetic aperture radar (InSAR) technology can provide a high-precision, wide-range monitoring of slow rock-slope deformation, making it an effective tool for studying geological hazards. Within the study area, multiple synthetic aperture radar (SAR) images from the Sentinel-1A satellite were collected, and surface deformation was obtained using the small baseline subset InSAR (SBAS-InSAR). The results demonstrate that combining ascending and descending orbit images can be successfully applied to landslide monitoring in complex mountainous areas. Over 30 potential landslides were identified by combining InSAR results with optical images. The Line-Of-Sight (LOS) direction deformation features and their relationship with precipitation were analyzed based on two typical landslides, and two-dimensional/three-dimensional (2D/3D) deformation decomposition was carried out to reveal its motion characteristics. It was found that the cumulative deformation fluctuation amplitude was higher during the rainy season, and the main movement direction of the landslide was east-west. In addition, based on the spatial distribution and statistical analysis of deformation points along with meteorological data, geological elements, human activities, and topographic conditions, it is inferred that factors such as low vegetation coverage, tectonic movements, human activities, and high-altitude glacier thawing may contribute to the occurrence of disasters. And it was found that areas with high vegetation cover, high rainfall, and snow cover exhibit lower coherence coefficients. This study offers valuable insights for investigating large-scale geological in alpine canyon regions.

Paxillin phase separation promotes focal adhesion assembly and integrin signaling.

Focal adhesions (FAs) are transmembrane protein assemblies mediating cell-matrix connection. Although protein liquid-liquid phase separation (LLPS) has been tied to the organization and dynamics of FAs, the underlying mechanisms remain unclear. Here, we experimentally tune the LLPS of PXN/Paxillin, an essential scaffold protein of FAs, by utilizing a light-inducible Cry2 system in different cell types. In addition to nucleating FA components, light-triggered PXN LLPS potently activates integrin signaling and subsequently accelerates cell spreading. In contrast to the homotypic interaction-driven LLPS of PXN in vitro, PXN condensates in cells are associated with the plasma membrane and modulated by actomyosin contraction and client proteins of FAs. Interestingly, non-specific weak intermolecular interactions synergize with specific molecular interactions to mediate the multicomponent condensation of PXN and are efficient in promoting FA assembly and integrin signaling. Thus, our data establish an active role of the PXN phase transition into a condensed membrane-associated compartment in promoting the assembly/maturation of FAs.