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The clinical importance of assessing and combining data on TP53mutations and isoforms is discussed in this article. It gives a succinct overview of the structural makeup and key biological roles of the isoforms. It then provides a comprehensive summary of the roles that p53 isoforms play in cancer development, therapy response and resistance. The review provides a summary of studies demonstrating the role of p53 isoforms as potential prognostic indicators. It further provides evidence on how the presence of TP53mutations may affect one or more of these activities and the association of p53 isoforms with clinicopathological data in various tumour types. The review gives insight into the present diagnostic hurdles for identifying TP53isoforms and makes recommendations to improve their evaluation. In conclusion, this review offers suggestions for enhancing the identification and integration of TP53isoforms in conjunction with mutation data within the clinical context.

To investigate the molecular mechanism and identify potential drugs for subthreshold depression (SD), and elucidate the detalied mechanism of Danzhi Xiaoyao powder (DZXY) in SD.

Kadsura coccinea has been used as a folk medicine in China, treating various diseases, including chronic gastritis and rheumatoid arthritis (RA). Chemical studies have revealed that lignans and triterpenes are the main constituents of its roots and stems. In this paper, a chemical investigation on the stem of K. coccinea was performed and a previously undescribed 3,4-secolanostane (1) was afforded. The structure of 1 was determined through comprehensive NMR and HRESI-MS data analysis. The stereo-configuration of 1 was established based on a comparison of experimental and calculated ECD spectra. Bioactivity assay carried out in immune cells indicated that 1 could show moderate effects on ConA (5 μg/mL)-induced T-cell proliferation with an IC50 value of 13.28 μM (cyclosporine A as the positive drug, IC50 = 0.13 μM) while the IC50 value was 23.12 μM in LPS (10 μg/mL)-induced B cells (mycophenolate mofetil as the positive drug, IC50 = 16.71 μM).

Background: The p53 isoform Δ133p53β is known to be associated with cancers driven by inflammation. Many of the features associated with the development of inflammation in rheumatoid arthritis (RA) parallel those evident in cancer progression. However, the role of this isoform in RA has not yet been explored. The aim of this study was to determine whether Δ133p53β is driving aggressive disease in RA. Methods: Using RA patient synovia, we carried out RT-qPCR and RNAScope-ISH to determine both protein and mRNA levels of Δ133p53 and p53. We also used IHC to determine the location and type of cells with elevated levels of Δ133p53β. Plasma cytokines were also measured using a BioPlex cytokine panel and data analysed by the Milliplex Analyst software. Results: Elevated levels of pro-inflammatory plasma cytokines were associated with synovia from RA patients displaying extensive tissue inflammation, increased immune cell infiltration and the highest levels of Δ133TP53and TP53βmRNA. Located in perivascular regions of synovial sub-lining and surrounding ectopic lymphoid structures (ELS) were a subset of cells with high levels of CD90, a marker of ‘activated fibroblasts’ together with elevated levels of Δ133p53β. Conclusions: Induction of Δ133p53β in CD90+synovial fibroblasts leads to an increase in cytokine and chemokine expression and the recruitment of proinflammatory cells into the synovial joint, creating a persistently inflamed environment. Our results show that dysregulated expression of Δ133p53β could represent one of the early triggers in the immunopathogenesis of RA and actively perpetuates chronic synovial inflammation. Therefore, Δ133p53β could be used as a biomarker to identify RA patients more likely to develop aggressive disease who might benefit from targeted therapy to cytokines such as IL-6.

To identify potential serum protein candidates involved in linking the traditional Chinese medicine (TCM)-defined qi deficiency constitution (QDC) to Pi-qi-deficiency syndrome (PQDS) of chronic superficial gastritis (CSG).

Determining the minimum supercooling temperature (Tsc) of unsaturated saline soils is the key scientific problem in cold‐region agricultural and engineering, and theoretical models with good predictability are urgently needed. This study establishes a framework for pore solution freezing nucleation in unsaturated saline soils, the chemical potential energy balance equation was established by determining phase transition driving force and potential energy barrier for the formation of stable crystal embryos. Finally, the physical model was constructed. By citing 46 sets of data from different researchers, the validity of the prediction results of this paper's model under different soil types and cooling rates is demonstrated, and the microscopic mechanism of supercooling occurring in the pore solution of positive permafrost is elucidated. The analysis found that electrolyte action restricts the free diffusion of molecules and promotes nucleation to occur, so that a maximum value of Tscexists during the growth of salinity (in NaCl saline soils occurs at a solution concentration of 0.26 mol/L). Too fast cooling rate will lead to a lag in crystallization time and an increase in supercooling. The combined water content determines the Tscwhen rc< 1 μm (pore size), and the effect of changes in water content and cooling rate on supercooling is more significant in small pores; the electrolyte concentration determines the Tscwhen rc> 1 μm, Tsc‐maxfrom concentration changes will be more significant in large pores. This study can provide theoretical reserves for controlling the freezing process of soils, studying the mechanism of freezing‐thawing hysteresis, and improving the related numerical calculation models. Permafrost and seasonal permafrost cover more than 50% of land area in the world. The engineering problems caused by freezing‐thawing cycles seriously affect engineering safety and people's well‐being in many countries. Therefore, an in‐depth study of the soil freezing‐thawing cycle is necessary. The water in the soil pore space remains sometimes in liquid state when it is below the freezing temperature. This phenomenon is usually referred to as supercooling. There is still a lack of quantitative studies on the minimum temperature that can be reached by this phenomenon. In this paper, a model that can reasonably predict the minimum subcooling temperature of soil is developed based on thermodynamic methods. The model is verified to be of good accuracy by means of measured data from different researchers. Subsequently, the supercooling characteristics of the soil are discussed based on the model and the microscopic mechanism of the gradual development of freezing in the supercooling state is described. This study will help to recognize and control the soil freezing process in engineering and agricultural construction, and serve to protect engineering structures. A physical model is proposed to quantify the supercooling degree of unsaturated saline soilsA maximum value of soil minimum supercooling temperature will occur with increasing salt concentrationToo fast cooling rate will lead to a lag in crystallization time and an increase in supercooling A physical model is proposed to quantify the supercooling degree of unsaturated saline soils A maximum value of soil minimum supercooling temperature will occur with increasing salt concentration Too fast cooling rate will lead to a lag in crystallization time and an increase in supercooling

The clarification of freezing in a soil‐water system is critical for assessing the formation of a freezing zone and liquid water flow. The supercooling phenomenon of soil pore solutions has been found during the freezing process, but the mechanism remains poorly understood. In this study, we propose a free energy function of soil‐water systems based on the Classical Nucleation Theory. The analytical solution of the critical nucleation problem of saline soil‐water system is obtained by combining the initial freezing temperature model and Pitzer activity coefficients model in electrolyte solutions. Then, the freezing‐thawing experiments of saline soil with various salt contents were conducted for verifying the analytical solution. The derived boundary nucleation rate is the quantitative solution of the critical condition for the supercooling. The findings suggested that the theory results agreed well with the experiment results. For the salt‐free soil‐water system, the critical maximum radius of supercooling was 7.15 nm. We compared eight classical ice‐water interfacial tension models, and the “Reinhardt & Doye” and “DeMott & Rogers” models showed excellent performance when using the new free energy theoretical framework to predict the crystallization nucleation rate of soil‐water systems. A positive correlation between the boundary nucleation rate and soil‐water potential is detected. According to the influencing factors, the boundary nucleation rate of soil‐water system can be divided into three zones: salt nature control zone (R> 100 μm), salt‐pore mixed control zone (100 μm > R> 100 nm), and pore size control zone (R< 100 nm). Unfrozen water refers to the soil moisture that still exists in the form of liquid water in the soil at negative temperatures, which is critical for determining the material properties of frozen soil. The phenomenon of soil water not being able to freeze as soon as the temperature falls below the freezing temperature is referred to as “supercooling.” However, the critical condition of supercooling is still unclear. In this paper, we developed a nucleation model that can be applied to saline soils based on the thermodynamic approach. Subsequently, the critical conditions were verified by freezing‐thawing tests of soil samples with different salt contents. We further optimized the model of this paper by selecting the best model for the intermediate variables. Quantitative relationships between the initiation condition of supercooling and soil properties were established. We also delineated pore size intervals for distinguishing differences in supercooling properties at different pore sizes. The proposed theoretical framework is developed from Classical Nucleation Theory, which may provide a theoretical reference for revealing the freezing‐thawing mechanism of soil‐water systems. There is supercooling in the freezing process and not supercooling‐jump phenomenon in the melting processThe boundary nucleation rate is proportional to the soil water potentialThere exists a critical pore size for the non‐saline soil where supercooling‐jump occurs, and the value is at the 10 nm level (7.15 nm) There is supercooling in the freezing process and not supercooling‐jump phenomenon in the melting process The boundary nucleation rate is proportional to the soil water potential There exists a critical pore size for the non‐saline soil where supercooling‐jump occurs, and the value is at the 10 nm level (7.15 nm)

The rapid pace of technological change in the internet age has fundamentally transformed information technology (IT) outsourcing. Thus, organisations should be able to re-imagine their IT outsourcing strategies to meet the needs of increasingly turbulent business environments. Bimodal IT requires the shaping of IT services to be either agile oriented or efficient oriented, thereby bringing new demands to an organisations' adoption strategies. On the basis of the review of traditional IT outsourcing service research, this paper presents the traditional and nonlinear modes of IT outsourcing service, discusses risk-based IT outsourcing service adoption strategy, relationship and contract management strategies from the static and dynamic perspectives and provides several directions for future research.

To screen for blood leukocyte microRNA (miRNA) biomarkers responsible for the association between the traditional Chinese medicine (TCM) qi deficiency constitution (QDC) and the Pi-qi-deficiency syndrome (PQDS) of chronic superficial gastritis (CSG).

Lymphatic capillaries develop discontinuous cell-cell junctions that permit the absorption of large macromolecules, chylomicrons, and fluid from the interstitium. While excessive vascular endothelial growth factor 2 (VEGFR2) signaling can remodel and seal these junctions, whether and how VEGFR3 can alter lymphatic junctions remains incompletely understood. Here, we use lymphatic-specific Flt4knockout mice to investigate VEGFR3 signaling in lymphatic junctions. We show that loss of Flt4prevents specialized button junction formation in multiple tissues and impairs interstitial absorption. Knockdown of FLT4in human lymphatic endothelial cells results in impaired NOTCH1 expression and activation, and overexpression of the NOTCH1 intracellular domain in Flt4knockout vessels rescues the formation of button junctions and absorption of interstitial molecules. Together, our data reveal a requirement for VEGFR3 and NOTCH1 signaling in the development of button junctions during postnatal development and may hold clinical relevance to lymphatic diseases with impaired VEGFR3 signaling.

The clustered regularly interspaced short palindromic repeat (CRISPR) gene editing technique, based on the non-homologous end-joining (NHEJ) repair pathway, has been used to generate gene knock-outs with variable sizes of small insertion/deletions with high efficiency. More precise genome editing, either the insertion or deletion of a desired fragment, can be done by combining the homology-directed-repair (HDR) pathway with CRISPR cleavage. However, HDR-mediated gene knock-in experiments are typically inefficient, and there have been no reports of successful gene knock-in with DNA fragments larger than 4 kb. Here, we describe the targeted insertion of large DNA fragments (7.4 and 5.8 kb) into the genomes of mouse embryonic stem (ES) cells and zygotes, respectively, using the CRISPR/HDR technique without NHEJ inhibitors. Our data show that CRISPR/HDR without NHEJ inhibitors can result in highly efficient gene knock-in, equivalent to CRISPR/HDR with NHEJ inhibitors. Although NHEJ is the dominant repair pathway associated with CRISPR-mediated double-strand breaks (DSBs), and biallelic gene knock-ins are common, NHEJ and biallelic gene knock-ins were not detected. Our results demonstrate that efficient targeted insertion of large DNA fragments without NHEJ inhibitors is possible, a result that should stimulate interest in understanding the mechanisms of high efficiency CRISPR targeting in general.

Restriction enzymes have two major limitations for cloning: they cannot cleave at any desired location in a DNA sequence and may not cleave uniquely within a DNA sequence. In contrast, the clustered regularly interspaced short palindromic repeat (CRISPR)–associated enzyme 9 (Cas9), when coupled with single guide RNAs (sgRNA), has been used in vivo to cleave the genomes of many species at a single site, enabling generation of mutated cell lines and animals. The Cas9/sgRNA complex recognizes a 17–20 base target site, which can be of any sequence as long as it is located 5′ of the protospacer adjacent motif (PAM; sequence 5′-NRG, where R = G or A). Thus, it can be programmed to cleave almost anywhere with a stringency higher than that of one cleavage in a sequence of human genome size. Here, the Cas9 enzyme and a specific sgRNA were used to linearize a 22 kb plasmid in vitro. A DNA fragment was then inserted into the linearized vector seamlessly through Gibson assembly. Our technique can be used to directly, and seamlessly, clone fragments into vectors of any size as well as to modify existing constructs where no other methods are available.

A true one-step one-pot aldol-reaction procedure has been developed for the synthesis of β-hydroxy ketones and esters. The reaction can be run at room temperature by simply mixing four components in CH2Cl2, with medium-to-high yields of aldol products obtained after regular workup. Mechanistically, the process probably proceeds via Mg-enolate formation of the ketone or ester component, followed by addition to the electrophilic aldehyde.

In this paper, we report a novel double-ended tuning fork (DETF) with variable sections. The DETF is fabricated using a quartz micro-electro-mechanical system (MEMS) technique and packaged using the flip chip technique. The central part of the vibration beam is thinned to enhance the force-frequency sensitivity but not decrease the vibration frequency. The mechanical quality factor ($Q$ value) can also be maintained, which determines the mechanical noise performance. Three types of DETF are designed and fabricated with a fixed beam length of 4 mm. The vibration characteristics (vibration frequency, $Q$ value, and equivalent circuit parameters) are evaluated using an impedance analyzer 4294A. The finite element method (FEM) is used to simulate the natural frequency and force-frequency sensitivity. For the natural frequencies, the experimental results agreed well with the simulation results. High $Q$ values are achieved for all the DETFs, which are 9924, 7083, and 6335. By multiplying the force-frequency sensitivity ($\Delta F$) by the measured $Q$ values, the performance of the new DETF structure can be improved by 1.9 times.