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Cancer remains a significant challenge in extending human life expectancy in the 21stcentury, with staggering numbers projected by the International Agency for Research on Cancer for upcoming years. While conventional cancer therapies exist, their limitations, in terms of efficacy and side effects, demand the development of novel treatments that selectively target cancer cells. Tumor immunotherapy has emerged as a promising approach, but low response rates and immune-related side effects present significant clinical challenges. Researchers have begun combining immunotherapy with nanomaterials to optimize tumor-killing effects. Stimuli-responsive nanomaterials have become a focus of cancer immunotherapy research due to their unique properties. These nanomaterials target specific signals in the tumor microenvironment, such as pH or temperature changes, to precisely deliver therapeutic agents and minimize damage to healthy tissue. This article reviews the recent developments and clinical applications of endogenous and exogenous stimuli-responsive nanomaterials for tumor immunotherapy, analyzing the advantages and limitations of these materials and highlighting their potential for enhancing the immune response to cancer and improving patient outcomes.

Purpose: Cross-sequence magnetic resonance image (MRI) registration and segmentation are two essential steps in a variety of medical image analysis tasks. And have attracted considerable research interest. However, they remain challenging due to domain shifts between different sequences. This study is aiming at proposing a novel method via disentangled representations, latent shape image learning (LSIL), for cross-sequence image registration and segmentation. Methods: Images from different sequences were firstly decomposed into a shared domain-invariant shape space and a domain-specific appearance space via an unsupervised image-to-image translation approach. A latent shape image learning model is then built on the disentangled shape representations to generate latent shape images. A series of experiments including cross-sequence image registration and segmentation were performed to qualitatively and quantitatively verify the validity of our method. Dice similarity coefficient (DSC) and 95th percentile Hausdorff distance (HD95) were adopted as our evaluation metrics. Results: The performance of our method was evaluated based on 2 datasets total of 50 MRIs. The experimental results showed the superiority of the proposed framework over the state-of-the-art cross-sequence registration and segmentation approaches. The proposed method shows the mean DSCs of 0.711 and 0.867, respectively, in cross-sequence registration and segmentation. Conclusion: We proposed a novel method based on representation disentangling to solve the cross-sequence registration and segmentation problem. Experimental results prove the feasibility and generalization of the generated latent shape images. The proposed method demonstrates significant potential for use in clinical environments of missing sequences. The source code is available at https://github.com/wujiong-hub/LSIL.

The proper regulation of transcription is essential for maintaining genome integrity and executing other downstream cellular functions1,2. Here we identify a stable association between the genome-stability regulator sensor of single-stranded DNA (SOSS)3and the transcription regulator Integrator-PP2A (INTAC)4–6. Through SSB1-mediated recognition of single-stranded DNA, SOSS–INTAC stimulates promoter-proximal termination of transcription and attenuates R-loops associated with paused RNA polymerase II to prevent R-loop-induced genome instability. SOSS–INTAC-dependent attenuation of R-loops is enhanced by the ability of SSB1 to form liquid-like condensates. Deletion of NABP2(encoding SSB1) or introduction of cancer-associated mutations into its intrinsically disordered region leads to a pervasive accumulation of R-loops, highlighting a genome surveillance function of SOSS–INTAC that enables timely termination of transcription at promoters to constrain R-loop accumulation and ensure genome stability.

Based on the proposed miniaturized polarization conversion metasurface (PCM), a checkerboard metasurface is designed, which can realize radar cross Section (RCS) reduction in the ultrawide range of 4.3–13.5 GHz (103%) by using the principle of reflection phase cancellation. The characteristic mode analysis (CMA) method is introduced to establish the checkerboard PCM as the radiation patch, and a wideband low-profile antenna for the C-band is achieved, with a bandwidth of 4.8–6 GHz (22.2%). The highlight and advantages are that the size of the checkerboard PCM is significantly reduced, and its side length is close to the radiation wavelength of the antenna. The antenna element based on this checkerboard PCM has the characteristics of small size and wide frequency range of RCS reduction, and the low-RCS antenna array can be realized by paving them seamlessly. Moreover, the checkerboard PCM not only suppresses scattering but also improves antenna performance and plays an important role in achieving wideband and enhancing radiation directivity.

Triple-negative breast cancer (TNBC) is a heterogeneous disease and lacks effective treatment. Our previous study classified TNBCs into four subtypes with putative therapeutic targets. Here, we report the final results of FUTURE, a phase II umbrella trial designed to explore whether the subtyping-based strategy may improve the outcomes in metastatic TNBC patients. A total of 141 patients with a median of three previous lines of therapies in the metastatic setting were enrolled in seven parallel arms. Confirmed objective responses were achieved in 42 patients (29.8%; 95% confidence interval [CI], 22.4–38.1). The median values of progression-free survival and overall survival were 3.4 (95% CI: 2.7–4.2) and 10.7 (95% CI: 9.1–12.3) months, respectively. Given Bayesian predictive probability, efficacy boundaries were achieved in four arms. Furthermore, integrated genomic and clinicopathological profiling illustrated associations of clinical and genomic parameters with treatment efficacy, and the efficacy of novel antibody–drug conjugates was explored in preclinical TNBC models of subtypes for which treatment was futile. In general, the FUTURE strategy recruits patients efficiently and provides promising efficacy with manageable toxicities, outlining a direction for further clinical exploration.

•DUSP4 enhances therapeutic efficacy in HER2-positive BC.•Elevated DUSP4 levels improved clinical outcomes in HER2-positive BC.•DUSP4 inhibits the ROS pathway.•DUSP4 hinders G6PD activity via ALDOB dephosphorylation.

Purpose: Aiming at the key issue of interpersonal interaction process between successors and top management teams in the context of family business trans-generational entrepreneurship, this paper aims to explore the motivation, action and results of interpersonal conflict between these two subjects mentioned above. Design/methodology/approach: This paper uses grounded theoretical research methods based on the research objects of three family businesses with both inheritance and transformation needs. Findings: Motivation difference, mediation mode and relationship utilization, that is, the successor and top management team take the goal-orientation, interest orientation and rational tendency as the starting point for relationship coordination. Then it mediates the interpersonal conflicts through compromise, collaboration and compliance, and ultimately provide successors with resources, opportunities, capabilities and motivation advantages to promote trans-generational entrepreneurship. Second, the path of relationship coordination comes from the background of social embedding, that is, the motivation difference comes from the embedding of motivation seeking, the mediation mode comes from the embedding of mediation elements and the relationship utilization comes from the embedding of relationship optimization. The research conclusions not only provide a theoretical framework for family businesses to solve the problems of interpersonal conflicts faced by family businesses but also have practical guiding significance for the trans-generational entrepreneurship. Originality/value: There are two theoretical contributions in this study. First, the research starting point of social embeddedness theory from the perspective of interpersonal relationships at the microlevel is revised. Since Granovetter (Granovetter, 1985) put forward the theory of social embeddedness, its research scope has been gradually expanded, but the mainstream research in the past focused on analyzing the social network embeddedness of enterprises to obtain social capital from the macro- and meso-level (Nahapiet and Ghoshal, 1998). In fact, this may deviate from the essential interpretation of Granovetter’s theory of social embeddedness, while this study returns to the study of interpersonal relationships. Second, a theoretical model of relationship coordination for successors and top management team is put forward in general. On the basis of the motivation, action and result of interpersonal interaction between the successor and top management team, the interpersonal coordination action path and embedded logic during trans-generational entrepreneurship of family businesses are revealed, which enriched the research scope of social embedded theory in family business.

IMPORTANCE: The value of platinum-based adjuvant chemotherapy in patients with triple-negative breast cancer (TNBC) remains controversial, as does whether BRCA1 and BRCA2 (BRCA1/2) germline variants are associated with platinum treatment sensitivity. OBJECTIVE: To compare 6 cycles of paclitaxel plus carboplatin (PCb) with a standard-dose regimen of 3 cycles of cyclophosphamide, epirubicin, and fluorouracil followed by 3 cycles of docetaxel (CEF-T). DESIGN, SETTING, AND PARTICIPANTS: This phase 3 randomized clinical trial was conducted at 9 cancer centers and hospitals in China. Between July 1, 2011, and April 30, 2016, women aged 18 to 70 years with operable TNBC after definitive surgery (having pathologically confirmed regional node-positive disease or node-negative disease with tumor diameter >10 mm) were screened and enrolled. Exclusion criteria included having metastatic or locally advanced disease, having non-TNBC, or receiving preoperative anticancer therapy. Data were analyzed from December 1, 2019, to January 31, 2020, from the intent-to-treat population as prespecified in the protocol. INTERVENTIONS: Participants were randomized to receive PCb (paclitaxel 80 mg/m2 and carboplatin [area under the curve = 2] on days 1, 8, and 15 every 28 days for 6 cycles) or CEF-T (cyclophosphamide 500 mg/m2, epirubicin 100 mg/m2, and fluorouracil 500 mg/m2 every 3 weeks for 3 cycles followed by docetaxel 100 mg/m2 every 3 weeks for 3 cycles). MAIN OUTCOMES AND MEASURES: The primary end point was disease-free survival (DFS). Secondary end points included overall survival, distant DFS, relapse-free survival, DFS in patients with germline variants in BRCA1/2 or homologous recombination repair (HRR)–related genes, and toxicity. RESULTS: A total of 647 patients (mean [SD] age, 51 [44-57] years) with operable TNBC were randomized to receive CEF-T (n = 322) or PCb (n = 325). At a median follow-up of 62 months, DFS time was longer in those assigned to PCb compared with CEF-T (5-year DFS, 86.5% vs 80.3%, hazard ratio [HR] = 0.65; 95% CI, 0.44-0.96; P = .03). Similar outcomes were observed for distant DFS and relapse-free survival. There was no statistically significant difference in overall survival between the groups (HR = 0.71; 95% CI, 0.42-1.22, P = .22). In the exploratory and hypothesis-generating subgroup analyses of PCb vs CEF-T, the HR for DFS was 0.44 (95% CI, 0.15-1.31; P = .14) in patients with the BRCA1/2 variant and 0.39 (95% CI, 0.15-0.99; P = .04) in those with the HRR variant. Safety data were consistent with the known safety profiles of relevant drugs. CONCLUSIONS AND RELEVANCE: These findings suggest that a paclitaxel-plus-carboplatin regimen is an effective alternative adjuvant chemotherapy choice for patients with operable TNBC. In the era of molecular classification, subsets of TNBC sensitive to PCb should be further investigated. TRIAL REGISTRATION: ClinicalTrials.gov Identifier: NCT01216111

The cadmium (Cd) pollution of farmland soil is serious in the world. The present study investigated the effects of intercropping Vicia fabaand the hyperaccumulator Sonchus asperon the Cd accumulation and root responses (morphology and secreted organic acids) of plants grown on soil from a mining area in Yunnan Province, China, under different Cd stress levels (0, 50, 100, and 200 mg kg–1). Intercropping increased the biomass of both S. asperand V. faba,as well as the Cd accumulation and Cd transfer coefficient from roots to shoots of S. asper,but decreased those of V. fabain the 200 mg kg–1Cd treatment. The Cd concentrations in roots, shoots, and grains from intercropped V. fabaplants were positively correlated (P <0.05) with the N,N-diethyl phenylacetamide-extractable Cd content in soil. In the 50 mg kg–1Cd treatment, intercropping decreased citric and malic acids in root secretions of S. asper.Intercropped V. fabasecreted more citric, oxalic, acetic, and malic acid compared with S. asper.Intercropping also increased root length and root surface area of both S. asperand V. faba.At 50 mg kg–1Cd, root length for S. asperwas higher than that at 0 mg kg–1Cd, whereas V. fabaroots had significantly decreased length and mean diameter. Intercropping of S. asperand V. fabais suggested as an in situphytoremediation strategy of Cd-contaminated soils and may improve the food quality of V. faba.

IMPORTANCE: During the past few decades, the incidence of breast cancer (BC) has been increasing rapidly in East Asia, and BC is currently the most common cancer in several countries. The rising incidence is likely related to changing lifestyle and environmental factors in addition to the increase in early diagnosis with BC awareness and screening. The understanding and management of BC are generally based on research and data from the West. However, emerging differences in BC epidemiology and tumor and host biology in Asian populations may be clinically relevant. OBSERVATIONS: A higher proportion of premenopausal BCs occur in Asia, although this factor is possibly an age-cohort effect. Although the relative frequencies of different immunohistochemical subtypes of BC may be similar between the East and West, the higher prevalence of luminal B subtypes with more frequent mutations in TP53 may be confounded by disparities in early detection. In addition, Asian BCs appear to harbor a more immune-active microenvironment than BCs in the West. The spectra of germline mutations in BC predisposition genes and single-nucleotide polymorphisms contributing to BC risk vary with ethnicity as well. Differences in tolerability of certain cytotoxic and targeted agents used in BC treatment may be associated with pharmacogenomic factors, whereas the lower body mass of the average woman in East Asia may contribute to higher toxicities from drugs administered at fixed doses. Phenotypic characteristics, such as lower breast volume, may influence the type of surgery performed in East Asian women. On the other hand, increased breast density may affect the sensitivity of mammography in detecting BCs, limiting the benefits of screening mammography. CONCLUSIONS AND RELEVANCE: Breast cancer has become a major health problem in Asia. The inclusion of more women from Asia in clinical trials and epidemiologic and translational studies may help unravel the interethnic heterogeneity of BCs and elucidate the complex interplay between environmental and intrinsic factors in its pathogenesis. These insights may help to refine prevention, diagnosis, and management strategies for BC in the setting of ethnic diversity.

Protein kinase B (PKB) has been implicated in the stimulation of glycogen synthesis by insulin in L6 muscle cells. This effect is dependent on phosphatidylinositol-3 kinase (PI-3 kinase), and mediated [...]

Antibodies labeled with positron-emitting isotopes have been used for tumor detection, predicting which patients may respond to tumor antigen-directed therapy, and assessing pharmacodynamic effects of drug interventions. Prolactin receptor (PRLR) is overexpressed in breast and prostate cancers and is a new target for cancer therapy. We evaluated REGN2878, an anti-PRLR monoclonal antibody, as an immunoPET reagent. REGN2878 was labeled with Zr-89 after conjugation with desferrioxamine B or labeled with I-131/I-124. In vitro determination of the half-maximal inhibitory concentration (IC50) of parental REGN2878, DFO-REGN2878, and iodinated REGN2878 was performed by examining the effect of the increasing amounts of these on uptake of trace-labeled I-131 REGN2878. REGN1932, a non-PRLR binding antibody, was used as a control. Imaging and biodistribution studies were performed in mice bearing tumor xenografts with various expression levels of PRLR, including MCF-7, transfected MCF-7/PRLR, PC3, and transfected PC3/PRLR and T4D7v11 cell lines. The specificity of uptake in tumors was evaluated by comparing Zr-89 REGN2878 and REGN1932, and in vivo competition compared Zr-89 REGN2878 uptake in tumor xenografts with and without prior injection of 2 mg of nonradioactive REGN2878. The competition binding assay of DFO-REGN2878 at ratios of 3.53–5.77 DFO per antibody showed IC50 values of 0.4917 and 0.7136 nM, respectively, compared to 0.3455 nM for parental REGN2878 and 0.3343 nM for I-124 REGN2878. Imaging and biodistribution studies showed excellent targeting of Zr-89 REGN2878 in PRLR-positive xenografts at delayed times of 189 h (presented as mean ± 1 SD, percent injected activity per mL (%IA/mL) 74.6 ± 33.8%IA/mL). In contrast, MCF-7/PRLR tumor xenografts showed a low uptake (7.0 ± 2.3%IA/mL) of control Zr-89 REGN1932 and a very low uptake and rapid clearance of I-124 REGN2878 (1.4 ± 0.6%IA/mL). Zr-89 REGN2878 has excellent antigen-specific targeting in various PRLR tumor xenograft models. We estimated, using image-based kinetic modeling, that PRLR antigen has a very rapid in vivo turnover half-life of ∼14 min from the cell membrane. Despite relatively modest estimated tumor PRLR expression numbers, PRLR-expressing cells have shown final retention of the Zr-89 REGN2878 antibody, with an uptake that appeared to be related to PRLR expression. This reagent has the potential to be used in clinical trials targeting PRLR.

Although therapeutic targets have been developed for colorectal cancer (CRC) therapy, the therapeutic effects are not ideal and the survival rate for CRC patients remains poor. Therefore, it is crucial to recognize a specific target and develop an efficacious delivery system for CRC therapy. Herein, we demonstrate that reduced ALKBH5 mediates aberrant m6A modification and tumor progression in CRC. Mechanically, histone deacetylase 2-mediated H3K27 deacetylation inhibits ALKBH5 transcription in CRC, whereas ectopic ALKBH5 expression decreases tumorigenesis of CRC cells and protects mice from colitis-associated tumor development. Further, METTL14/ALKBH5/IGF2BPs combine to modulate JMJD8 stability in an m6A-dependent manner, which increases glycolysis and accelerates the development of CRC by enhancing the enzymatic activity of PKM2. Moreover, ALKBH5 mRNA-loaded folic acid-modified exosome–liposome hybrid nanoparticles were synthesized and significantly inhibit the progression of CRC in preclinical tumor models by modulating the ALKBH5/JMJD8/PKM2 axis and inhibiting glycolysis. Overall, our research confirms the crucial function of ALKBH5 in regulating the m6A status in CRC and provides a direct preclinical approach for using ALKBH5 mRNA nanotherapeutics for CRC.

Video Discussion by Karol A. Gutowski, M.D., is Available Online for this Article.

This paper presents a discrete element method to handle particle collision detection and responses in transport simulation (the simulation of transport of protons and neutrons in granular flow target geometric model) based on GPUs. Discrete element method was adopted in the realisation of large-scale particle visualisation. The method simulates and solves edge detection, position judging, motion direction, calculation of the next collision point using GPU acceleration during the process of transport, and demonstrates the complete interaction process through OpenGL. Results show that the model presented exploits the acceleration of GPUs and has gained remarkable functional improvement compared with traditional method using solely CPUs. In addition, we used the MCNPX to calculate this model with high-speed proton bombardment. The distribution of power energies verifies that the granular flow target model is reliable and feasible.

We investigate, simulate, and measure triple-band electromagnetically induced transparency-like metamaterial with different shapes and sizes of split-ring resonators. It exhibits three high transmissivities at 8.19 GHz, 8.65 GHz, and 9.19 GHz which can be fitted by the equivalent circuit model. Meanwhile, this has high values of group delay at three transparency peaks, exhibiting an excellent slow-light effect. On the other hand, this is applied to design the multi-band refractive index sensor, exhibiting a favorable sensing property. The physics mechanism of transparency peaks is analyzed by the electric field distributions.

This paper is states that training the students’ ability, combine with the course characters of the C programming language, and from the system of practice teaching, advance design of the content, the methods of practice teaching practice to do the deep exploration, the teaching effect is significant .

To determine the efficacy and toxicity of an oxaliplatin-based regimen as a neoadjuvant chemotherapy setting in triple-negative local advanced breast cancer (TNLABC) patients.

Dual-Fano resonances have been proposed with all-dielectric toroidal metamaterials in the microwave frequency. The presented structure consists of high permittivity rectangular bars of equal spacing. Two transparency peaks derived from the strong toroidal dipole with two modes can reach 0.92 and 0.9 at 10.51 GHz and 16.27 GHz, respectively. The excitation of these resonances could be explained by near-field distribution and multipoles scattered power. The trend of experimental results is virtually uniform with that of theoretical analysis and simulated curves. Such a compact all-dielectric structure may have important implications in many fields, such as ultra-sensitive sensors, stealth transmission, and so on.

To examine the relationship between large tumor size and breast cancer–specific mortality (BCSM), especially in a subset of patients with negative lymph nodes (LNs).

Due to the present condition of temperature examination in laser molten pool which is mainly limited to infrared thermometer and thermal infrared imager ,a temperature measurement system based on monochrome CCD with high speed is developed. By transforming gray value to temperature , grayscale images of molten pool will be changed into 2 dimension distribution with color, making examination more intuitive and comprehensive .

The research use an impact test enginery, choose T10 steel samples, and do a low stress repeated impact collisions experiment. By comparing two kinds of different heat treatment of T10 steel, analyze and research its macro plastic deformation phenomena and plastic accumulation law in low stress repeated collision impact load, and observe its hardness change by hardness microscopicand and its metallographic structure by metallographic microscope to discuss its deformation causes.

The aim of this study was to establish a differential high-resolution melting analysis for the detection of K-rascodons 12 and 13 mutations in pancreatic cancer tissue.

Due to the small volume and high density of breast tissue in Asian women, particularly younger women, mammographic diagnosis is sometimes non-conclusive, with a Breast Imaging Reporting and Data System (BI-RADS) result of 0. No alternative based on blood biomarkers has yet succeeded in discriminating between patients with breast cancer (BC) and those with benign breast disease (BBD) among BI-RADS 0 patients. In our study, 84 BC and 94 BBD patients with mammographic results and confirmed pathologic information were enrolled and categorized into two groups, namely, 79 BC and 73 BBD patients with BI-RADS 1-5 and 5 BC and 21 BBD patients with BI-RADS 0. RNA extracted from peripheral blood samples collected in PAXgeneTMtubes was analyzed after NuGEN WT-OvationTMRNA amplification using Affymetrix GeneChip

Aims and background To investigate the clinicopathological characteristics and prognosis of breast cancer subtypes classified by quantitative estrogen receptor (ER), progesterone receptor (PR), and Her2.Methods and study design 923 patients with primary breast cancer having a median age of 53 years who were treated at the Cancer Hospital of Fudan University in Shanghai between January 2002 and June 2004 were retrospectively analyzed. Four molecular subtypes were constructed from the immunohistochemical results of quantitative hormone receptor (HR) and Her2 status. HR+ was defined as ER+ and PR+, HR± as ER/PR+ at lower levels or lacking either ER or PR, and HR- as both ER- and PR-. The four subtypes were HR+/Her2-, HR±/Her2-, HR-/Her2- (triple-negative), and Her2+. Clinical and pathological parameters, disease-free survival (DFS), and overall survival (OS) measurements were compared between patients with different molecular subtypes.Results The proportions of HR+/Her2-, HR±/Her2-, triple-negative, and Her2+ breast cancer were 36.6% (338/923), 22.9% (211/923), 20.6% (190/923), and 19.9% (194/923). The median follow-up was 49.0 months (4–77 months). In 145 cases disease recurrence or death occurred. In multivariate analysis with the HR+/Her2- subtype taken as the reference category, triple-negative and Her2+ subtypes were associated with increased recurrence and death with a hazard ratio (HR) of 2.05 (95% CI 1.31–3.20; P = 0.002) and 1.89 (95% CI 1.20–2.97, P = 0.006) for DFS and 2.84 (95% CI 1.45–5.55; P = 0.002) and 2.95 (95% CI 1.51–5.77, P = 0.002) for OS, respectively; the HR±/Her2- subtype was marginally associated with poor prognosis with HR 1.51 (95% CI 0.94–2.43; P = 0.088) and 1.90 (95% CI 0.92–3.94; P = 0.084) for DFS and OS, respectively.Conclusions Breast cancer subtypes based on quantitative ER, PR, and Her2 may be predictive of prognosis. Patients whose tumors were not HR+/Her2- had a worse outcome in our study.

Backgrounds Hemodynamics plays an important role in the natural history of the process of rupture and recurrence of intracranial aneurysms. This study aimed to investigate the role of hemodynamics for recurrence in a vertebral artery dissecting aneurysm (VADA).Methods A patient with a ruptured VADA firstly treated by low-profile visualized intraluminal support (LVIS)-assisted coiling, and was implanted with a Pipeline Embolization Device (PED) after aneurysm recurrence. Finite element analysis and computational fluid dynamics simulations were conducted in 6 serial imaging procedures, and the calculated hemodynamics was correlated with aneurysm recurrence.Results Wall shear stress (WSS) was not effectively suppressed, resulting in aneurysm recurrence with initial entry tear to occur above the protuberance after 7 months of LVIS stent-assisted coiling. With the implantation of PED, WSS, inflow stream and velocity at the aneurysm neck significantly decreased. During the 3-month follow-up after PED deployment, there was significant shrinkage of the sac and the blood flow in the sac was reduced considerably. The 27-month follow-up after PED deployment indicated the aneurysm was stable.Conclusions The present case study suggests that insufficient suppression of high WSS and high inflow velocity at the neck of the parent artery, especially near the posterior inferior cerebellar artery, might be associated with aneurysm recurrence.

Aims and background To evaluate the immunohistochemical characterization of CK5/6 and CK17 and whether the expression level of the two markers was correlated with clinical outcome or pathological feature in triple negative (ER-, PR-, HER-2-) patients with breast cancer.Methods and study design We carried out an immunohistochemical assay for CK5/6 and CK17 markers on formalin-fixed invasive carcinoma samples from 112 patients who were diagnosed between 2000 and 2002. All of them had an immunohistochemical triple negative status and follow-up information available.Results Of the 112 patients characterized by triple negative immunohistochemical status, 82 (73.2%) were disease free with no relapse or metastasis. In total, CK5/6 and CK17 were both determined positive in 33.9% (38/112) of the 112 tumor samples, and 46.4% (52/112) were regarded as positive for CK5/6 or CK17. The Kaplan-Meier curve showed that positive staining for CK5/6, CK17, or CK which means CK5/6 positive or CK17 positive, was associated with worse disease-free survival (P = 0.020, P = 0.032, P = 0.003), and positive staining for CK5/6 or CK was associated with worse overall survival (P = 0.027, P = 0.015). When we considered 91 patients whose pathological type was invasive ductal carcinoma, we found that there was also an association between CK5/6 or CK17 immunostaining and high grade (P = 0.030). In addition, these two markers were also associated with axillary lymph node status (P = 0.044). The Cox regression multiple-factor analysis showed that pathological stage, grade and expression of CK were the factors affecting both disease-free and overall survival, whereas age and menopausal status were independent factors affecting disease-free and overall survival, respectively.Conclusions Positive staining for CK5/6 or CK17 was associated with a worse prognosis, high tumor grade and positive axillary lymph nodes.

The molecular mechanisms involved in breast cancer metastasis still remain unclear to date. In our previous study, differential expression of peroxiredoxin 6 was found between the highly metastatic MDA-MB-435HM cells and their parental counterparts, MDA-MB-435 cells. In this study, we investigated the effects of peroxiredoxin 6 on the proliferation and metastatic potential of human breast cancer cells and their potential mechanism. Expression of peroxiredoxin 6 in the highly metastatic MDA-MB-231HM cells was investigated by RT-PCR, real-time PCR and western blot. A recombinant expression plasmid of the human peroxiredoxin 6 gene was constructed and transfected into MDA-MB-231 and MDA-MB-435 cells. The effects of peroxiredoxin 6 on the proliferation and invasion of MDA-MB-231 and MDA-MB-435 cells were investigated by the Cell Counting Kit-8 method, colony-formation assay, adhesion assay, flow cytometry and invasion assay in vitro. miRNA was used to downregulate the expression of peroxiredoxin 6. Genes related to the invasion and metastasis of cancer were determined by RT-PCR, real-time PCR and western blot. The tumorigenicity and spontaneously metastatic capability regulated by peroxiredoxin 6 were determined using an orthotopic xenograft tumor model in athymic mice. Overexpression of peroxiredoxin 6 in MDA-MB-231HM cells compared with their parental counterparts was confirmed. Upregulation of peroxiredoxin 6 enhanced the in vitroproliferation and invasion of breast cancer cells. The enhancement was associated with decreasing levels of tissue inhibitor of matrix metalloproteinase (TIMP)-2 and increasing levels of the urokinase-type plasminogen activator receptor (uPAR), Ets-1 (E26 transformation-specific-1), matrix metalloproteinase (MMP)-9 and RhoC (ras homolog gene family, member C) expression. The results were further demonstrated by RNA interference experiments in vitro. In an in vivostudy, we also demonstrated that peroxiredoxin 6-transfected breast cancer cells grew much faster and had more pulmonary metastases than control cells. By contrast, peroxiredoxin 6 knockdown breast cancer cells grew more slowly and had fewer pulmonary metastases. Effects similar to those of peroxiredoxin 6 on the uPAR, Ets-1, MMP-9, RhoC and TIMP-2 expression observed in in vitrostudies were found in the in vivostudy. Overexpression of peroxiredoxin 6 leads to a more invasive phenotype and metastatic potential in human breast cancer, at least in part, through regulation of the levels of uPAR, Ets-1, MMP-9, RhoC and TIMP-2 expression.

Abstract: Objective: To discuss the suitable immediate breast reconstruction modalities for Chinese patients by comparing the pedicled transverse rectus abdominis myocytaneous flap (TRAM) reconstruction with latissimus dorsi myocytaneous flap (LTD) reconstruction plus implants or not after mastectomy due to breast cancer. Methods: From Jan. 2000 to Jul. 2005, 74 staged 0-II patients (mean age 39) were performed immediate breast reconstruction with autologous tissue either using LTD flaps or pedicled TRAM flaps with supplemental implants when necessary after mastectomy due to breast cancer and the charts were reviewed. Results: The age, marriage and menses status did not affect the selection of modalities and the need of implants. In 74 patients, 62 cases (83.8%) were performed LTD reconstruction with 13 implants and 12 cases received TRAM with 1 implant. The difference in need of implants or not between the two modalities had no statistical significance (P=0.442, Fisher’ exact test). Aesthetic results judged as good or fair were in 88% patients and the cosmetic effects between LTD and TRAM groups or implant and non-implant groups had no differences. All reconstructions were successful, with 4.1% cumulative locoregional recurrence and 100% overall survival by following up to 66 months (median 9 months). The DFS and RFS between the two modalities had no significant differences by log rank test. Conclusion: Immediate autologous tissue reconstruction makes it possible to regain the natural and symmetric contour of breast without increased local recurrence. The LTD flap reconstruction is a suitable option for most Chinese women as well as the pedicled TRAM flap.

Twenty-five to thirty percent of patients with node-negative breast cancer are expected to relapse following surgery, therefore great efforts have been made to identify new prognostic markers that could be useful in defining patients for additional therapy. The expression of MMP-2 and MMP-9 has been associated with high potential of metastasis in several human carcinomas including breast cancer. In the present study we examined the prognostic value of immunoreactive MMP-2/MMP-9 protein in 270 consecutive lymph node negative cases who received radical mastectomy or modified radical mastectomy. Among the patients, 211 cases received adjuvant endocrine therapy and/or adjuvant chemotherapy. Using immunohistochemical assay, we found that 56.7% of the resected tumors were positive for MMP-2 whereas 59.6% of the samples were positive for MMP-9. χ2test demonstrated a significant direct association between MMP-2 and MMP-9 ( p< 0.001); positive immunostaining of MMP-2 was significantly related to higher tumor grade ( p< 0.001) and larger tumor size ( p=0.012); positive immunostaining of MMP-9 was significantly related to higher tumor grade ( p=0.002). In univariate analysis, using Cox-proportional hazard model we found MMP-2, MMP-9 and the co-expression of MMPs (MMP2/MMP9) were significantly associated with patients' relapse free survival ( p=0.016, 0.015 and 0.013 respectively) but not overall survival ( p=0.122, 0.320 and 0.091 respectively). Log-rank test also showed that MMP-2, MMP-9 or the co-expression of MMP2/MMP9 was unfavorable prognostic factor for relapse free survival but not overall survival. In subgroup analysis, we found MMPs were more prognostic for patients with no adjuvant treatment than for patients with adjuvant therapy. In multivariate analysis, using Cox-proportional hazard model we found co-expression of MMPs, larger tumor size and higher tumor grade were unfavorable for relapse free survival ( p=0.038, 0.007 and 0.015 for each). We concluded that MMP-2 and MMP-2 are unfavorable prognostic factors in breast cancer patients. They might be potential predictive factor for adjuvant systemic therapy. The co-expression of MMP-2 and MMP-9 has significantly prognostic value in node-negative patients.

ABSTRACTInsulin receptor substrate 1 (IRS-1) plays an important role in the insulin signaling cascade. In vitro and in vivo studies from many investigators have suggested that lowering of IRS-1 cellular levels may be a mechanism of disordered insulin action (so-called insulin resistance). We previously reported that the protein levels of IRS-1 were selectively regulated by a proteasome degradation pathway in CHO/IR/IRS-1 cells and 3T3-L1 adipocytes during prolonged insulin exposure, whereas IRS-2 was unaffected. We have now studied the signaling events that are involved in activation of the IRS-1 proteasome degradation pathway. Additionally, we have addressed structural elements in IRS-1 versus IRS-2 that are required for its specific proteasome degradation. Using ts20 cells, which express a temperature-sensitive mutant of ubiquitin-activating enzyme E1, ubiquitination of IRS-1 was shown to be a prerequisite for insulin-induced IRS-1 proteasome degradation. Using IRS-1/IRS-2 chimeric proteins, the N-terminal region of IRS-1 including the PH and PTB domains was identified as essential for targeting IRS-1 to the ubiquitin-proteasome degradation pathway. Activation of phosphatidylinositol 3-kinase is necessary but not sufficient for activating and sustaining the IRS-1 ubiquitin-proteasome degradation pathway. In contrast, activation of mTOR is not required for IRS-1 degradation in CHO/IR cells. Thus, our data provide insight into the molecular mechanism of insulin-induced activation of the IRS-1 ubiquitin-proteasome degradation pathway.

Time-resolved Förster resonance energy transfer (trFRET) has been used to obtain interdye distance distributions. These distributions give the most probable distance as well as a parameter, σ, that characterize the width of the distribution. This latter parameter contains information not only on the flexibility of the dyes tethered to macromolecules, but on the flexibility of the macromolecules. Both the most probable interdye distance as well as σ provide insight into DNA static bending and DNA flexibility. Time-resolved fluorescence anisotropy and static anisotropy measurements can be combined to provide a measure of the cone angle within which the tethered dyes appear to wobble. When this motion is an order of magnitude faster than the average lifetime that characterizes transfer, an average value of the dipolar orientational parameter κ² can be calculated for various mutual dye orientations. The resulting κ² distribution is very much narrower than the limiting values of 0 and 4, allowing more precise distances and distance changes to be determined. Static and time-resolved fluorescence data can be combined to constrain the analyses of DNA–protein kinetics to provide thermodynamic parameters for binding and for conformational changes along a reaction coordinate. The parameter σ can be used to model multiple DNA–protein complexes with varying DNA bend angles in a global fitting of trFRET data. Such a global fitting approach has shown how the range of bends in single base DNA variants, when bound by the TATA binding protein (TBP), can be understood in terms of two limiting forms. Time-resolved FRET, combined with steady-state FRET, can be used to show not only how osmolytes affect the binding of DNA to proteins, but also how DNA bending depends on osmolyte concentration in the DNA–protein complexes. © 2002 Wiley Periodicals, Inc. Biopoly (Nucleic Acid Sci) 61: 180–200, 2002

The TATA-binding protein (TBP) initiates assembly of transcription preinitiation complexes on eukaryotic class II promoters, binding to and restructuring consensus and variant “TATA box” sequences. The sequence dependence of the DNA structure in TBP·TATA complexes has been investigated in solution using fluorescence resonance energy transfer. The mean 5′dye-3′dye distance varies significantly among oligomers bearing the adenovirusmajor late promoter sequence (AdMLP) and five single-site variants bound to Saccharomyces cerevisiaeTBP, consistent with solution bend angles for AdMLP of 76° and for the variants ranging from 30° to 62°. These solution bends contrast sharply with the corresponding co-crystal structures, which show ∼80° bends for all sequences. Transcription activities for these TATA sequences are strongly correlated with the solution bend angles but not with TBP·DNA binding affinities. Our results support a model in which transcription efficiency derives primarily from the sequence-dependent structure of the TBP·TATA binary complex. Specifically, the distance distribution for the average solution structure of the TBP·TATA complex may reflect the sequence-dependent probability for the complex to assume a conformation in which the TATA box DNA is severely bent. Upon assumption of this geometry, the binary complex becomes a target for binding and correctly orienting the other components of the preinitiation complex.

The complex formed between the TATA-binding protein (TBP) and the “TATA box” of eukaryotic class II promoters is the foundation for assembly of the complex to which RNA polymerase II is ultimately recruited. TBP binds productively to canonical and diverse variant TATA sequences with >100-fold differences in transcription efficiency. Co-crystals of canonical sequences and >11 variant sequences bound to various TBP molecules all have ∼80° bends. In contrast, the bend angles for TBP·TATA complexes in solution, derived from distance distributions, are ∼80° for a canonical sequence but range from 30° to 62° for five variant sequences (1). We show in this study that the osmolytes used to crystallize TBP·TATA complexes induce profound increases in the DNA bends of two transcriptionally active TBP-bound variant sequences to a common angle of ∼80° but have little effect on a transcriptionally inactive variant. The effect of osmolyte on the TBP-induced DNA bend of a variant TATA box sequence is also manifest in the kinetics of association, demonstrating a functional consequence of an osmolyte-induced structural change.

In previous work we showed that the phosphatidylinositol 3-kinase (PI3-kinase), not the mitogen-activated protein kinase, pathway is necessary and sufficient to account for insulin- and epidermal growth factor (EGF)-induced DNA synthesis in rat hepatocytes. Here, using a dominant-negative p85, we confirmed the key role of EGF-induced PI3-kinase activation and sought to identify the mechanism by which this is effected. Our results show that EGF activates PI3-kinase with a time course similar to that of the association of p85 with three principal phosphotyrosine proteins (i.e.PY180, PY105, and PY52). We demonstrated that each formed a distinct p85-associated complex. PY180 and PY52 each constituted about 10% of EGF-activated PI3-kinase, whereas PY105 was responsible for 80%. PY105 associated with Grb2 and SHP-2, and although it behaved like Gab1, none of the latter was detected in rat liver. We therefore cloned a cDNA from rat liver, which was found to be 95% homologous to the mouse Grb2-associated binder 2 (Gab2) cDNA sequence. Using a specific Gab2 antibody, we demonstrated its expression in and association with p85, SHP-2, and Grb2 upon EGF treatment of rat hepatocytes. Gab2 accounted for most if not all of the PY105 species, since immunoprecipitation of Gab2 with specific antibodies demonstrated parallel immunodepletion of Gab2 and PY105 from the residual supernatants. We also found that the PI3-kinase activity associated with Gab2 was totally abolished by dominant negative p85. Thus, Gab2 appears to be the principal EGF-induced PY protein recruiting and activating PI3-kinase and mitogenesis.

ABSTRACTHistone (de)acetylation is important for the regulation of fundamental biological processes such as gene expression and DNA recombination. Distinct classes of histone deacetylases (HDACs) have been identified, but how they are regulated in vivo remains largely unexplored. Here we describe results demonstrating that HDAC4, a member of class II human HDACs, is localized in the cytoplasm and/or the nucleus. Moreover, we have found that HDAC4 interacts with the 14-3-3 family of proteins that are known to bind specifically to conserved phosphoserine-containing motifs. Deletion analyses suggested that S246, S467, and S632 of HDAC4 mediate this interaction. Consistent with this, alanine substitutions of these serine residues abrogated 14-3-3 binding. Although these substitutions had minimal effects on the deacetylase activity of HDAC4, they stimulated its nuclear localization and thus led to enhanced transcriptional repression. These results indicate that 14-3-3 proteins negatively regulate HDAC4 by preventing its nuclear localization and thereby uncover a novel regulatory mechanism for HDACs.

Objective: To study the prognostic effect of bcl-2 oncogene and its gene family members bax, bcl-x expression in breast cancer patients. Methods: Expression of bcl-2, bax proteins in 91 human breast cancer tissue sections were studied by immunohistochemical method, bcl-x1 mRNA expression in frozen tissues from 16 breast cancer patients were detected using Northern blot method. Results: bcl-2 protein positivity was found in 60/91 (65.9%) patients, and bax positivity 59/91 (64.8%). bcl-2 and bax expression levels were associated with apoptotic index(AI), histological grade, axillary lymph node metastasis, postoperative local recurrence and metastasis, bcl-2 expression was related to ER positivity. In univariate analysis for disease free survival (DFS), bcl-2 and bax protein levels, and Al were all found to have prognostic value. The result of Cox’s model multivariate analysis showed that bcl-2 protein level was an independent prognostic factor. In 16 frozen breast cancer tissues, 8/16(50%) had higher level of bcl-x1 mRNA, which showed correlation with bcl-2 protein expression and axillary lymph node metastasis. Conclusion: The findings indicate that dysregulated expressions of bcl-2, bax and bcl-x1 apoptosis-related genes, suggestive of serious deregulation of apoptotic process, may contribute to the biologic aggressiveness of breast cancer, bcl-2 protein is an independent indicator of prognosis in breast cancer patients.

De-escalating anthracycline is gaining popularity for breast cancer patients. We aim to evaluate the non-inferiority of an anthracycline-free or short-term regimen to the standard anthracycline-based regimen for operable patients with human epidermal growth factor receptor 2 (HER2)-negative breast cancer.

Proteins which are secreted or associated with the cell envelope of Mycobacterium tuberculosismay contain protective T‐cell epitopes. Prior to this study, a recombinant clone bank of enzymatically active M. tuberculosis‐alkaline phosphatase fusions, were screened for immunogenicity in a murine T‐cell model. Five of these were selected for further study, and the IFN‐γ secretion and proliferation of human PBMC from purified protein derivative‐ (PPD)‐positive and PPD‐negative donors were measured in response to oligopeptides, Mtb‐PhoA fusions and one full‐length protein. Epitopes from four of the five selected antigens were immunoreactive in the human model and corresponded to cytochrome dubiquinol oxidase, cytochrome coxidase subunit II, MTV005.02 and MTV033.08. Thus, this strategy identified novel human immunogenic peptides as possible candidates for a subunit vaccine.

The sgpgene from Streptococcus mutanshas been previously isolated, characterized, and demonstrated to encode a G‐protein. In order to investigate the function of this gene, a novel antisense RNA strategy was developed. Expression of sgpantisense RNA in Escherichia coliled to transient inhibition of growth. In addition, sgpantisense RNA expression in S. mutansresulted in decreased growth under environmental stress conditions (44°C, acidic pH, and high osmolarity). Therefore, these results suggest that the sgpgene plays a role in modulating the stress responses of S. mutans. This approach could be applicable for investigating the function of essential genes in other organisms for which mutants are not available.

Abstract: Objective: This study was undertaken to determine the expression of apoptosis in breast cancer and to evaluate its significance as a prognostic marker. Methods: A series of 91 invasive breast cancer was analysed for the expression of apoptosis by using the 3-end-labeling method of DNA in tissue sections. The apoptotic indexes were the percentages of apoptotic cells among tumor cells. Results: The end-labeling method allowed a precise evaluation of the expression of apoptosis. Apoptosis occurred in 91.1% of breast cancer patients, and apoptotic indexes were divided into two groups, 0–0.21 and 0.28–0.62. Low apoptotic index was related to axillary lymph node metastasis (P<0.01). In survival analysis, higher apoptotic index was related to disease free survival (P=0.0095) and overall survival (P=0.0348) in the entire cohort. Cox’s analysis showed that apoptotic index had no independent prognostic value. Conclusion: The apoptosis was a spontaneous phenomenon in breast cancer tissue, and the expression was different from each other. Further analysis was needed to clarify the relationship between apoptosis and prognosis, especially the response to adjuvant therapy.