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Summary: Recapitulating spontaneous metastasis in preclinical models is crucial for understanding mechanisms underlying cancer progression and testing effective therapeutic interventions. We present a protocol for establishing and characterizing the spontaneous metastasis model in mice. We describe steps for generating primary tumors, tumor resection, monitoring metastatic dissemination, and evaluating metastatic burden using histological and imaging techniques. This protocol provides a valuable tool for studying metastasis in vivo and testing therapeutic strategies aimed at preventing or targeting metastatic diseases.For complete details on the use and execution of this protocol, please refer to Liu et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Abstract Distinguishing indolent from clinically significant localized prostate cancer is a major clinical challenge and influences clinical decision-making between treatment and active surveillance. The development of novel predictive biomarkers will help with risk stratification, and clinical decision-making, leading to a decrease in over or under-treatment of patients with prostate cancer. Here, we report that Trop2 is a prognostic tissue biomarker for clinically significant prostate cancer by utilizing the Canary Prostate Cancer Tissue Microarray (CPCTA) cohort composed of over 1100 patients from a multi-institutional study. We demonstrate that elevated Trop2 expression is correlated with worse clinical features including Gleason score, age, and pre-operative PSA levels. More importantly, we demonstrate that elevated Trop2 expression at radical prostatectomy predicts worse overall survival in men undergoing radical prostatectomy. Additionally, we detect shed Trop2 in urine from men with clinically significant prostate cancer. Our study identifies Trop2 as a novel tissue prognostic biomarker and a candidate non-invasive marker for prostate cancer.

Summary: Lysine crotonylation has attracted widespread attention in recent years. However, little is known about bacterial crotonylation, particularly crotonyltransferase and decrotonylase, and its effects on antibiotic resistance. Our study demonstrates the ubiquitous presence of crotonylation in E. coli, which promotes bacterial resistance to polymyxin. We identify the crotonyltransferase YjgM and its regulatory pathways in E. coli with a focus on crotonylation. Further studies show that YjgM upregulates the crotonylation of the substrate protein PmrA, thereby boosting PmrA’s affinity for binding to the promoter of eptA, which, in turn, promotes EptA expression and confers polymyxin resistance in E. coli. Additionally, we discover that PmrA’s crucial crotonylation site and functional site is Lys 164. These significant discoveries highlight the role of crotonylation in bacterial drug resistance and offer a fresh perspective on creating antibacterial compounds.

Currently, the quest for bamboo materials with high color fastness, rich colors and environmental friendliness is rapidly rising due to its potential applications in construction, furniture and decoration. However, finding an easy-to-operate and environmentally friendly dye for bamboo is a necessary task because of the difficulty in treating the dyeing waste liquid of acid dyes and the complexity of the production process of reactive dyes.Five formulations involving metal polyphenol complexes were employed to straightforwardly produce eco-friendly dyed bamboo and the impact of various formulations on the light aging resistance of the dyed veneers was examined. The results indicated that the light resistance of bamboo veneer dyed with the solution containing only FeSO4·7H2O and tannic acid reached level 4, surpassing the undyed bamboo veneer by three levels. The mechanism of enhanced lightfastness of dyed bamboo veneer was elucidated by XPS analysis. The polyphenol iron complex serves a dual purpose: it absorbs ultraviolet rays and scavenges free radicals within the system. Additionally, it reduced the oxidation of phenolics in the substrate, transforming them into dark-colored quinone structures. This process enhanced the light-aging resistance of the finishing materials. Therefore, this work provides a simple and environmentally friendly method for changing the color of bamboo and provides a new idea for the selection of dyes for bamboo dyeing in actual production.

Abstract BRCA1 deficient breast cancers are aggressive and chemoresistant due, in part, to their enrichment of cancer stem cells that can be generated from carcinoma cells by an epithelial-mesenchymal transition (EMT). We previously discovered that BRCA1 deficiency activates EMT in mammary tumorigenesis. How BRCA1 controls EMT and how to effectively target BRCA1-deficient cancers remain elusive. We analyzed murine and human tumors and identified a role for Tgfβr2 in governing the molecular aspects of EMT that occur with Brca1 loss. We utilized CRISPR to delete Tgfβr2 and specific inhibitors to block Tgfβr2 activity and followed up with the molecular analysis of assays for tumor growth and metastasis. We discovered that heterozygous germline deletion, or epithelia-specific deletion of Brca1 in mice, activates Tgfβr2 signaling pathways in mammary tumors. BRCA1 depletion promotes TGFβ-mediated EMT activation in cancer cells. BRCA1 binds to the TGFβR2 locus to repress its transcription. Targeted deletion or pharmaceutical inhibition of Tgfβr2 in Brca1-deficient tumor cells reduces EMT and suppresses tumorigenesis and metastasis. BRCA1 and TGFβR2 expression levels are inversely related in human breast cancers. This study reveals for the first time that a targetable TGFβR signaling pathway is directly activated by BRCA1-deficiency in the induction of EMT in breast cancer progression.

Abstract Rapid advances in flexible and wearable smart textiles demand low-cost, high-energy/power-density fiber-shaped supercapacitors (FSCs). The performance of FSCs is determined by the fabrication and assembly of fiber-shaped electrodes (FSEs), where an active charge-storage material is always clad around flexible charge transmission current collectors. Inspired by the tissue structure of natural bamboo, wherein parenchyma cells (PCs) that store nutrients are clad around bamboo fibers (BFs), we propose a strategy for converting bamboo cells into FSEs using conductive BFs and activated PCs as current collectors and active materials, respectively. The assembled electrode has a high specific capacitance of 1454 mF cm−2 at 0.64 mA cm−2. A solid-state FSC with a pair of bamboo-structured electrodes exhibited a substantially high energy density. Its mechanical flexibility enabled the knitting of wearable wristbands to drive ultra-small voltmeter indicators. This lightweight, low cost, and high-energy-density bamboo-structured FSC could enable numerous smart textile applications.

ABSTRACT Antibiotic resistance is increasingly becoming a challenge to public health. The regulation of bacterial metabolism by post-translational modifications (PTMs) has been widely studied. However, the mechanism underlying the regulation of acetylation in bacterial resistance to antibiotics is still unknown. Here, we performed a quantitative analysis of the acetylated proteome of a wild-type (WT) Escherichia coli (E. coli) sensitive strain and ampicillin- (Re-Amp), kanamycin- (Re-Kan), and polymyxin B-resistant (Re-Pol) strains. Based on bioinformatics analysis combined with biochemical validations, we found a common regulatory mechanism between the different resistant strains. Our results showed that protein acetylation negatively regulates bacterial metabolism to regulate antibiotic resistance and positively regulates bacterial motility. Further analyses revealed that key enzymes in various metabolic pathways were differentially acetylated. In particular, pyruvate kinase (PykF), a glycolytic enzyme that regulates bacterial metabolism, and its acetylated form were highly expressed in the three resistant strains and were identified as reversibly acetylated by the deacetylase CobB and the acetyl-transferase PatZ (peptidyl-lysine N-acetyltransferase). Results showed that PykF also could be acetylated by nonenzymatic acetyl phosphatase (AcP) in vitro. Furthermore, the deacetylation of Lys413 in PykF increased PykF enzymatic activity by changing the conformation of its ATP binding site, resulting in an increase in energy production which, in turn, increased the sensitivity of drug-resistant strains to antibiotics. This study provides novel insights for understanding bacterial resistance and lays the foundation for future research on the regulation of acetylation in antibiotic-resistant strains. IMPORTANCE The misuse of antibiotics has resulted in the emergence of many antibiotic-resistant strains which seriously threaten human health. Protein post-translational modifications, especially acetylation, tightly control bacterial metabolism. However, the comprehensive mechanism underlying the regulation of acetylation in bacterial resistance remains unexplored. Here, acetylation was found to positively regulate bacterial motility and negatively regulate energy metabolism, which was common in all antibiotic-resistant strains. Moreover, the acetylation and deacetylation process of PykF was uncovered, and deacetylation of the Lys 413 in PykF was found to contribute to bacterial sensitivity to antibiotics. This study provides a new direction for research on the development of bacterial resistance through post-translational modifications and a theoretical basis for developing antibacterial drugs.

Background: Peri-menopausal syndrome (PMPS) has a high incidence rate and seriously affects the physical and mental health of women. Honghua Xiaoyao Pill (HHXYP) is a Chinese patent medicine, which has been reported to be used to treat PMPS. However, there is still a lack of randomized clinical trial to evaluate the efficacy and safety of HHXYP on life quality, mood and vasomotor symptoms for PMPS women. This study aims to investigate whether HHXYP is effective and safe in treating PMPS and the possible mechanism.Methods: A multicenter, randomized, controlled clinical trial will be conducted in China to evaluate the efficacy and safety of HHXYP. Sixty women with peri-menopausal syndrome will be recruited at three centers and randomly in a 1:1 ratio to a treatment group using HHXYP (HHXYP group) and a control group using oryzanol (OC group). Participants will be treated with HHXYP or oryzanol for 12 weeks and followed up for 4 weeks. The primary outcome is the modified Kupperman Index (KI), which will be measured at baseline and 4, 8, 12, 16 weeks after randomization. The secondary outcomes include Hot flash scale (HFs), Menopause-Specific Quality of Life Scale (MENQOL) and Hamilton Depression/Anxiety Scale (HAMD/HAMA). The HFs are measured at the same point as the KI, other secondary outcomes are measured at baseline and 12, 16 weeks after randomization. The other outcomes are the levels of serum sex hormone, monoamine neurotransmitter, vascular vasomotor factor and the expression of phosphatidylinositol 3-active enzyme (PI3K)/protein activator enzyme B (Akt), which will be measured at baseline and 12 weeks after randomization. Adverse events will also be reported.Discussion: HHXYP is a potential alternative Chinese patent medicine for PMPS. This trial will provide evidence for HHXYP on improving the quality of life, mood and vasomotor symptoms, and sex hormone levels of PMPS patients.

Abstract Breast cancer remains the second most lethal cancer among women in the United States and triple-negative breast cancer is the most aggressive subtype with limited treatment options. Trop2, a cell membrane glycoprotein, is overexpressed in almost all epithelial cancers. In this study, we demonstrate that Trop2 is overexpressed in triple-negative breast cancer (TNBC), and downregulation of Trop2 delays TNBC cell and tumor growth supporting the oncogenic role of Trop2 in breast cancer. Through proteomic profiling, we discovered a metabolic signature comprised of TALDO1, GPI, LDHA, SHMT2, and ADK proteins that were downregulated in Trop2-depleted breast cancer tumors. The identified oncogene-mediated metabolic gene signature is significantly upregulated in TNBC patients across multiple RNA-expression clinical datasets. Our study further reveals that the metabolic gene signature reliably predicts poor survival of breast cancer patients with early stages of the disease. Taken together, our study identified a new five-gene metabolic signature as an accurate predictor of breast cancer outcome.

Abstract Neuroendocrine prostate cancer (NEPC) is a lethal subtype of prostate cancer that rarely develops de novo in primary tumors and is commonly acquired during the development of treatment resistance. NEPC is characterized by gain of neuroendocrine markers and loss of androgen receptor (AR), making it resistant to current therapeutic strategies targeting the AR signaling axis. Here, we report that MCM2, MCM3, MCM4, and MCM6 (MCM2/3/4/6) are elevated in human NEPC and high levels of MCM2/3/4/6 are associated with liver metastasis and poor survival in prostate cancer patients. MCM2/3/4/6 are four out of six proteins that form a core DNA helicase (MCM2-7) responsible for unwinding DNA forks during DNA replication. Inhibition of MCM2-7 by treatment with ciprofloxacin inhibits NEPC cell proliferation and migration in vitro, significantly delays NEPC tumor xenograft growth, and partially reverses the neuroendocrine phenotype in vivo. Our study reveals the clinical relevance of MCM2/3/4/6 proteins in NEPC and suggests that inhibition of MCM2-7 may represent a new therapeutic strategy for NEPC.

Abstract Prostate cancer remains the most common non-cutaneous malignancy among men in the United States. To discover potential serum-based biomarkers for high-risk prostate cancer, we performed a high-multiplex immunoassay utilizing patient-matched pre-operative and post-operative serum samples from ten men with high-grade and high-volume prostate cancer. Our study identified six (CASP8, MSLN, FGFBP1, ICOSLG, TIE2 and S100A4) out of 174 proteins that were significantly decreased after radical prostatectomy. High levels of CASP8 were detected in pre-operative serum samples when compared to post-operative serum samples and serum samples from patients with benign prostate hyperplasia (BPH). By immunohistochemistry, CASP8 protein was expressed at higher levels in prostate cancer tissues compared to non-cancerous and BPH tissues. Likewise, CASP8 mRNA expression was significantly upregulated in prostate cancer when compared to benign prostate tissues in four independent clinical datasets. In addition, mRNA levels of CASP8 were higher in patients with recurrent prostate cancer when compared to patients with non-recurrent prostate cancer and high expression of CASP8 was associated with worse disease-free survival and overall survival in renal cancer. Together, our results suggest that CASP8 may potentially serve as a biomarker for high-risk prostate cancer and possibly renal cancer.

Abstract Background Basal-like breast cancers (BLBCs) are a leading cause of cancer death due to their capacity to metastasize and lack of effective therapies. More than half of BLBCs have a dysfunctional BRCA1. Although most BRCA1-deficient cancers respond to DNA-damaging agents, resistance and tumor recurrence remain a challenge to survival outcomes for BLBC patients. Additional therapies targeting the pathways aberrantly activated by BRCA1 deficiency are urgently needed. Methods Most BRCA1-deficient BLBCs carry a dysfunctional INK4-RB pathway. Thus, we created genetically engineered mice with Brca1 loss and deletion of p16 INK4A , or separately p18INK4C , to model the deficient INK4-RB signaling in human BLBC. By using these mutant mice and human BRCA1-deficient and proficient breast cancer tissues and cells, we tested if there exists a druggable target in BRCA1-deficient breast cancers. Results Heterozygous germline or epithelium-specific deletion of Brca1 in p18INK4C - or p16 INK4A -deficient mice activated Pdgfrβ signaling, induced epithelial-to-mesenchymal transition, and led to BLBCs. Confirming this role, targeted deletion of Pdgfrβ in Brca1-deficient tumor cells promoted cell death, induced mesenchymal-to-epithelial transition, and suppressed tumorigenesis. Importantly, we also found that pharmaceutical inhibition of Pdgfrβ and its downstream target Pkcα suppressed Brca1-deficient tumor initiation and progression and effectively killed BRCA1-deficient cancer cells. Conclusions Our work offers the first genetic and biochemical evidence that PDGFRβ-PKCα signaling is repressed by BRCA1, which establishes PDGFRβ-PKCα signaling as a therapeutic target for BRCA1-deficient breast cancers.

Abstract Background Primary dysmenorrhea (PD) is a leading cause of dysmenorrhea among adolescent girls. Manual acupuncture may be considered as an effective treatment for PD, but high-quality evidence remains limited. This trial aims to evaluate the efficacy and safety of acupuncture for the prevention of PD as compared with sham acupuncture and usual care. Methods/design This is a three-arm, randomized, controlled clinical trial in which the patients, assessors, and statisticians will be blinded. A total of 300 acupuncture-naive patients who were diagnosed as PD will be randomly allocated to the verum acupuncture, sham acupuncture, or usual care groups in a 2:2:1 ratio. Patients in the verum acupuncture group will receive manual acupuncture at specific acupuncture points with penetrating needling, while those in the sham acupuncture group will receive non-penetrating needling at non-acupuncture points. They will be given five sessions over a menstrual cycle for 3 menstrual cycles. Patients in the usual care group will receive health education and informed to receive manual acupuncture for free after waiting for 7 menstrual cycles. The primary outcome will be the change from baseline in the Cox Menstrual Symptom Scale Score (CMSS). The secondary outcomes will be the changes in Massachusetts General Hospital Acupuncture Sensation Scale (MASS), visual analog scale (VAS), Short-Form McGill Pain Questionnaire 2 (SF-MPQ-2), Pittsburgh Sleep Quality Index (PSQI), Beck Anxiety Inventory (BAI), Beck Depression Inventory II (BDI- II), Acupuncture Expectancy Scale (AES), 60-item NEO Personality Inventory-Short Form (NEO-FFI), and acute medication intake. The adverse events will be recorded at every visit. The analyses will be performed base on a full analysis set (FAS) and a per-protocol set (PPS). Discussion This study may provide high-quality evidence regarding the efficacy and safety of manual acupuncture for PD. In addition, the results of this study will help to identify the efficacy of acupuncture due to the specific effects of acupuncture or placebo effects of acupuncture ritual. Trial registration Clinical Trials.gov NCT02783534 . Registered on 26 May 2016

Under the uncertain statistical framework by Liu [19], there is still a lack of an effective fitting method for uncertain linear models with Box-Cox transformation of response variables. For example, for the transformation parameter Λ , the uncertain least squares estimation will produce a severely low estimation result. In this paper, we propose uncertain Box-Cox regression analysis by utilizing the uncertainty theory to model the imprecise data and applying a generalized Box-Cox transformation indexed by its parameter to validate classic regression assumptions. We use rescaled least squares to estimate unknown parameters and provide an estimate for noises followed by residual analysis for these uncertain Box-Cox regression models. We also give the forecast values and confidence intervals and use a numerical example to demonstrate our methodology. Our work sets a uniformed framework for Box-Cox transformation on the uncertain regression, and extends such regression from linear to nonlinear cases, taking the Johnson-Schumacher growth model as an example.

We propose and numerically demonstrate a secure optical communication scheme in a small-world semiconductor lasers network based on cluster chaos synchronization and false-message scrambling. According to the symmetries of network, the semiconductor lasers are divided into sets of heterogeneous coupled clusters. The properties of cluster chaos synchronization, the performance and security of optical chaotic communication, as well as the influence of false message on the synchronization of clusters are systematically investigated. It is demonstrated that, by properly setting the parameters of the lasers network, high-quality chaos synchronization can be achieved between the intra-cluster lasers, whereas incoherent states are observed between inter-cluster lasers. Moreover, by inserting false messages into the inter-cluster couplings, the privacy of chaotic carriers transmitted over the inter-cluster public links can be significantly enhanced, as such secure bidirectional communication among intra-cluster lasers is achievable with a message bit rate of several Gbit/s. Compared with the conventional point-to-point communication systems, the proposed scheme supports multipoint-to-multipoint secure communications.

Wood-based products manufactured from fast-growing wood species such as eucalyptus have gained increasing attraction with the demand of using wood in architecture, furniture, and decoration. In this paper, a new type of wood scrimber based on eucalyptus veneers complexed with ferrous ions was prepared and its properties were characterized. The results showed that the presence of complexes did not affect the mechanical properties of eucalyptus wood scrimber, but made its surface more hydrophobic (contact angle increased by 38.48% and dimensional stability improved (thickness swelling rate decreased by 32.26%). Most importantly, the color of eucalyptus wood scrimber changed significantly, from the original brown to dark blue, and its anti-photoaging property also greatly improved. These advantages would make this type of wood scrimber based on the eucalyptus veneer complexes with ferrous ions more widely applicable in decorations and buildings.

Regression analysis is to establish the regression relationship between dependent variables and independent variables. The data of traditional regression model are often assumed to be observed precisely. However this assumption holds only sometimes. Due to the influence of various uncertain factors, the data in reality is often inaccurate. Therefore, We treat real data as uncertain variables. Uncertain regression analysis is likely to provide an effective analysis method. Based on the uncertainty theory, the residual analysis of verhulst-pearl model is discussed in this paper. We use the least square method to estimate the parameters. And we also obtained the confidence interval of the response variables for the new explanatory variables. In the uncertain regression analysis, we propose a leave-p-out cross-validation method for model selection under imprecise observation. We end up the paper with a numerical example of uncertain Verhulst-Pearl regression model and show that the model has a better prediction accuracy.

Background: Insomnia is a prevalent sleep disorder and strong risk factor for poor quality of life, depression, and other lifestylerelated diseases. Objectives: To investigate the effect of the transcutaneous vagus nerve stimulation (taVNS) on sleep electroencephalographic (EEG) in the para-chlorophenylalanine (PCPA) insomnia rats. Methods: Rats were divided into control, model, taVNS and sham taVNS (stnVNS) (stimulate the auricular margin, as transcutaneous none VNS, stnVNS) group (n=6 in each group). A week before the experiment, the electrodes were fixed to the skull of all the rats for recording the sleep EEG. PCPA was used to establish insomnia models. The rats of taVNS and stnVNS group were treated via an electroacupuncture apparatus for seven consecutive days, and simultaneously, the sleep EEG were recorded for all groups after the treatment daily. And the power spectrum analysis was used in this experiment. Results: After modeling, the percentage of power spectrum of delta frequency band significantly decreased, while the theta, alpha, and beta frequency bands significantly increased in the model group compared to the control group. After intervention, the percentage of the delta frequency band significantly increased in the taVNS group as compared to the stnVNS group. Conclusion: These results suggest that taVNS can significantly modulate the power spectrum of the delta frequency band and may constitute a potential low-cost alternative for the treatment of insomnia.

Hyperspectral image (HSI) analysis has become one of the most active topics in the field of remote sensing, which could provide powerful assistance for sensing a larger-scale environment. Nevertheless, a large number of high-correlation and redundancy bands in HSI data provide a massive challenge for image recognition and classification. Hybrid Rice Optimization (HRO) is a novel meta-heuristic, and its population is approximately divided into three groups with an equal number of individuals according to self-equilibrium and symmetry, which has been successfully applied in band selection. However, there are some limitations of primary HRO with respect to the local search for better solutions and this may result in overlooking a promising solution. Therefore, a modified HRO (MHRO) based on an opposition-based-learning (OBL) strategy and differential evolution (DE) operators is proposed for band selection in this paper. Firstly, OBL is adopted in the initialization phase of MHRO to increase the diversity of the population. Then, the exploitation ability is enhanced by embedding DE operators into the search process at each iteration. Experimental results verify that the proposed method shows superiority in both the classification accuracy and selected number of bands compared to other algorithms involved in the paper.

Uncertain differential equations (UDEs) with jumps are an essential tool to model the dynamic uncertain systems with dramatic changes. The interest rates, impacted heavily by human uncertainty, are assumed to follow UDEs with jumps in ideal markets. Based on this assumption, two derivatives, namely, interest-rate caps (IRCs) and interest-rate floors (IRFs), are investigated. Some formulas are presented to calculate their prices, which are of too complex forms for calculation in practice. For this reason, numerical algorithms are designed by using the formulas in order to compute the prices of these structured products. Numerical experiments are performed to illustrate the effectiveness and efficiency, which also show the prices of IRCs are strictly increasing with respect to the diffusion parameter while the prices of IRFs are strictly decreasing with respect to the diffusion parameter.

CAR (chimeric antigen receptor) T cell therapy has shown clinical success in treating hematological malignancies, but its treatment of solid tumors has been limited. One major challenge is on-target, off-tumor toxicity, where CAR T cells also damage normal tissues that express the targeted antigen. To reduce this detrimental side-effect, Boolean-logic gates like AND-NOT gates have utilized an inhibitory CAR (iCAR) to specifically curb CAR T cell activity at selected nonmalignant tissue sites. However, the strategy seems inefficient, requiring high levels of iCAR and its target antigen for inhibition. Using a TROP2-targeting iCAR with a single PD1 inhibitory domain to inhibit a CEACAM5-targeting CAR (CEACAR), we observed that the inefficiency was due to a kinetic delay in iCAR inhibition of cytotoxicity. To improve iCAR efficiency, we modified three features of the iCAR--the avidity, the affinity, and the intracellular signaling domains. Increasing the avidity but not the affinity of the iCAR led to significant reductions in the delay. iCARs containing twelve different inhibitory signaling domains were screened for improved inhibition, and three domains (BTLA, LAIR-1, and SIGLEC-9) each suppressed CAR T function but did not enhance inhibitory kinetics. When inhibitory domains of LAIR-1 or SIGLEC-9 were combined with PD-1 into a single dual-inhibitory domain iCAR (DiCARs) and tested with the CEACAR, inhibition efficiency improved as evidenced by a significant reduction in the inhibitory delay. These data indicate that a delicate balance between CAR and iCAR signaling strength and kinetics must be achieved to regulate AND-NOT gate CAR T cell selectivity. [ABSTRACT FROM AUTHOR]