Pharmacological targeting PIKfyve and tubulin as an effective treatment strategy for double-hit lymphoma.

Double-hit lymphoma is one of the most aggressive and refractory lymphoma subtypes with recurrent genetic abnormalities of MYC and BCL-2 or BCL6 rearrangement, leading to a poor prognosis in the present clinical practice. Therefore, new therapeutic strategies for eliminating double-hit lymphomas are urgently needed. Here, we reported that HZX-02-059, a novel PIKfyve and tubulin dual-target inhibitor, showed a highly cytotoxic activity against double-hit lymphoma cell lines in vitro and in vivo through a noncanonical caspase-independent cell death, methuosis. Mechanistically, the cytotoxicity triggered by HZX-02-059 was contributed to the PIKfyve/TFEB axis-induced cell death of methuosis, as well as the inhibition of tubulin and mTOR/Myc axis-induced cell cycle arrest. In summary, the present findings suggest that HZX-02-059 represents a good starting point for developing targeted therapeutics against double-hit lymphomas.

A new ALK inhibitor overcomes resistance to first- and second-generation inhibitors in NSCLC.

More than 60% of nonsmall cell lung cancer (NSCLC) patients show a positive response to the first ALK inhibitor, crizotinib, which has been used as the standard treatment for newly diagnosed patients with ALK rearrangement. However, most patients inevitably develop crizotinib resistance due to acquired secondary mutations in the ALK kinase domain, such as the gatekeeper mutation L1196M and the most refractory mutation, G1202R. Here, we develop XMU-MP-5 as a new-generation ALK inhibitor to overcome crizotinib resistance mutations, including L1196M and G1202R. XMU-MP-5 blocks ALK signaling pathways and inhibits the proliferation of cells harboring either wild-type or mutant EML4-ALK in vitro and suppresses tumor growth in xenograft mouse models in vivo. Structural analysis provides insights into the mode of action of XMU-MP-5. In addition, XMU-MP-5 induces significant regression of lung tumors in two genetically engineered mouse (GEM) models, further demonstrating its pharmacological efficacy and potential for clinical application. These preclinical data support XMU-MP-5 as a novel selective ALK inhibitor with high potency and selectivity. XMU-MP-5 holds great promise as a new therapeutic against clinically relevant secondary ALK mutations.

Nigericin is effective against multidrug resistant gram-positive bacteria, persisters, and biofilms.

Multidrug-resistant (MDR) bacteria pose a significant clinical threat to human health, but the development of antibiotics cannot meet the urgent need for effective agents, especially those that can kill persisters and biofilms. Here, we reported that nigericin showed potent bactericidal activity against various clinical MDR Gram-positive bacteria, persisters and biofilms, with low frequencies of resistance development. Moreover, nigericin exhibited favorable in vivo efficacy in deep-seated mouse biofilm, murine skin and bloodstream infection models. With Staphylococcus aureus, nigericin disrupted ATP production and electron transport chain; cell death was associated with altered membrane structure and permeability. Obtaining nigericin-resistant/tolerant mutants required multiple rounds of challenge, and, cross-resistance to members of several antimicrobial classes was absent, probably due to distinct nigericin action with the GraSR two-component regulatory system. Thus, our work reveals that nigericin is a promising antibiotic candidate for the treatment of chronic or recurrent infections caused by Gram-positive bacteria.

The qualitative accuracy of clinical dermatophytes via matrix-assisted laser desorption ionization-time of flight mass spectrometry: A meta-analysis.

Dermatophytes are an important part of superficial fungal infections, and accurate diagnosis is paramount for successful treatment. Recently, matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) has emerged as a powerful tool to identify clinical pathogens; its advantages are cost-effectiveness, rapid detection, and high accuracy. However, as the accurate identification of clinical dermatophytes via MALDI-TOF MS has still not been fully evaluated, we performed a meta-analysis for its systematic evaluation. Fifteen eligible studies were involved and showed high accuracy with an identification ratio of 0.96 (95% CI = 0.92─1.01) and 0.91 (95% CI = 0.86─0.96) at the genus and species levels, respectively. The results showed higher accuracy ratio of Vitek MS (91%) than MALDI Biotyper (85%). Dermatophytes such as Trichophyton interdigitale (0.99, 95% CI = 0.97─1.02), T. mentagrophytes var interdigitale (1.00, 95% CI = 0.98─1.02), and Microsporum canis (0.97, 95% CI = 0.89─1.04) showed high accuracy in detected clinical dermatophytes. Moreover, a library with self-built database set up by laboratories showed higher accuracy than commercial database, and 15-day cultivation for dermatophytes showed highest accuracy considering culture time. High heterogeneity was observed and decreased only with the subgroup analysis of species. The subgroup analysis of mass spectrometry, library database, and culture time also exhibited high heterogeneity. In summary, our results showed that MALDI-TOF MS could be used for highly accurate detection of clinically pathogenic dermatophytes, which could be an alternative diagnostic method in addition to morphological and molecular methods.

This meta-analysis comprehensively investigated the qualitative accuracy of clinical dermatophytes through MALDI-TOF MS. Owing to the high accuracy observed at both genus and species levels, this approach could be an alternative diagnostic method in addition to morphological and molecular methods.

Laboratory investigation of permeability property of natural gravels with different particle sizes under different laying conditions.

The seepage properties of natural gravel are one of the problems to be considered in seepage project designs. In this paper, the seepage properties of the natural gravel with particle sizes of 5, 20 and 60 mm were investigated under different laying conditions. The effect of the particle size, laying depth, bulk density and pressurized head on the seepage properties of the natural gravel was analyzed by using the combined methods of theoretical analysis with physical model test. The results showed that the seepage flow in the natural gravel was non-laminar flow in the test conditions described in this paper. Meanwhile, the relationship between particle size, laying depth, bulk density, pressurized heads and seepage property was established. The seepage discharge increased with the increase of the pressurized head and particle size, and decreased with the increasing of laying depth and bulk density. The critical laying depth and bulk density can be obtained when the seepage discharge becomes zero. The empirical formula of the seepage discharge of natural gravel with different particle sizes, laying depths, bulk densities and pressurized heads was obtained with the method of nonlinear regression, which can be expressed as: Q=5.9546d0.3713γ-0.2974L-0.1156H0.1307-5.6614. The empirical formula was experimentally validated. The maximum relative error did not exceed 6.73%, proving that the empirical formula of the seepage discharge of natural gravel was rational. The results can provide an important reference to further studying the seepage properties of macropore media, and form a theoretical basis for applying the natural gravel in the seepage projects.

Study on the velocity characteristics of spiral flow in pipeline with different numbers of guide bars installed on the surface of the piped vehicle body.

The piped hydraulic transportation of tube-contained raw material is a new low-carbon and environmental protection technique for transporting materials. The velocity characteristics of the spiral flow in the pipe with different numbers of guide bars installed on the surface of the piped vehicle body during piped hydraulic transportation of tube-contained raw material were studied by the theoretical analysis and model test. In the test, the numbers of guide bars placed on the surface of the piped vehicle body was respectively 3, 4, 5, and 6, and the studied sections were the section of the piped vehicle body and its rear section, and the flow discharge was 40 m3/h. The results showed that as the number of guide bars increased, the axial velocity of the section of the piped vehicle body increased gradually, while the axial velocity of the rear section of the piped vehicle reduced first and then increased. The section circumferential and radial flow velocity of the piped vehicle body and its rear section both increased first and then reduced. When the number of guide bars installed on the surface of the piped vehicle body was 4, the section circumferential flow velocity of the piped vehicle body and its rear section reached the maximum value, and their distributions were relatively uniform. The results offer theoretical basis so that we can optimize the structure of the piped vehicle and further popularize the piped hydraulic transportation technique of tube-contained raw material.

Discovery and Identification of Small Molecules as Methuosis Inducers with in Vivo Antitumor Activities.

Methuosis is a novel nonapoptotic mode of cell death characterized by vacuole accumulation in the cytoplasm. In this article, we describe a series of azaindole-based compounds that cause vacuolization in MDA-MB-231 cells. The most potent vacuole inducer, compound 13 (compound 13), displayed differential cytotoxicities against a broad panel of cancer cell lines, such as MDA-MB-231, A375, HCT116, and MCF-7, but it did not inhibit the growth of the nontumorigenic epithelial cell line MCF-10A. A mechanism study confirmed that the cell death was caused by inducing methuosis. Furthermore, compound 13 exhibited substantial pharmacological efficacy in the suppression of tumor growth in a xenograft mouse model of MDA-MB-231 cells without apparent side effects, which makes this compound the first example of a methuosis inducer with potent in vivo efficacy. These results demonstrate that methuosis inducers might serve as novel therapeutics for the treatment of cancer.

Kinetics and thermodynamics of urea hydrolysis under the coupling of nitrogen application rate and temperature.

This study aimed to determine the coupled effects of temperature and urea application rate on kinetic and thermodynamic parameters to supplement the mechanism of urea hydrolysis and modify the Arrhenius model to improve the prediction accuracy of urea content. Laboratory experiments were conducted for sandy loam soil under different temperatures (T) (288, 293, 298, and 308 K) and urea application rates (F) (247, 309, 371, and 433 mg kg-1). Urea content was determined daily through high-performance liquid chromatography. Results showed that the interaction between temperature and urea application rate had a significant effect on reaction rate (Ku) and half-life (H1/2), whereas no significant effect on activation degree (lgN), activation free energy (ΔG), activation enthalpy (ΔH), and activation entropy (ΔS). The new Ku(T)-2 model with a determination coefficient (R2) = 0.990 was more accurate than the Arrhenius model with R2 = 0.965. The new U(T, F) model with a mean absolute percentage error (MAPE) = 3.62% was more accurate than the traditional U(T) model with a MAPE = 6.38%. The effects of T and F were observed mainly during the preparatory stage and the most critical transition stage of the chemical reaction, respectively. The findings ΔH > 0, ΔG > 0, and ΔS < 0 indicated that urea hydrolysis was endothermic and controlled by enthalpy. These results supplemented the mechanism of urea hydrolysis and improved the prediction accuracy of urea content.

Prediction of soil urea conversion and quantification of the importance degrees of influencing factors through a new combinatorial model based on cluster method and artificial neural network.

Quantitative prediction of soil urea conversion is crucial in determining the mechanism of nitrogen transformation and understanding the dynamics of soil nutrients. This study aimed to establish a combinatorial prediction model (MCA-F-ANN) for soil urea conversion and quantify the relative importance degrees (RIDs) of influencing factors with the MCA-F-ANN method. Data samples were obtained from laboratory culture experiments, and soil nitrogen content and physicochemical properties were measured every other day. Results showed that when MCA-F-ANN was used, the mean-absolute-percent error values of NH4+-N, NO3--N, and NH3 contents were 3.180%, 2.756%, and 3.656%, respectively. MCA-F-ANN predicted urea transformation under multi-factor coupling conditions more accurately than traditional models did. The RIDs of reaction time (RT), electrical conductivity (EC), temperature (T), pH, nitrogen application rate (F), and moisture content (W) were 32.2%-36.5%, 24.0%-28.9%, 12.8%-15.2%, 9.8%-12.5%, 7.8%-11.0%, and 3.5%-6.0%, respectively. The RIDs of the influencing factors in a descending order showed the pattern RT > EC > T > pH > F > W. RT and EC were the key factors in the urea conversion process. The prediction accuracy of urea transformation process was improved, and the RIDs of the influencing factors were quantified.

Mycoepoxydiene suppresses HeLa cell growth by inhibiting glycolysis and the pentose phosphate pathway.

Upregulation of glycolysis and the pentose phosphate pathway (PPP) is a major characteristic of the metabolic reprogramming of cancer and provides cancer cells with energy and vital metabolites to support their rapid proliferation. Targeting glycolysis and the PPP has emerged as a promising antitumor therapeutic strategy. Marine natural products are attractive sources for anticancer therapeutics, as evidenced by the antitumor drug Yondelis. Mycoepoxydiene (MED) is a natural product isolated from a marine fungus that has shown promising inhibitory efficacy against HeLa cells in vitro. We used a proteomic approach with two-dimensional gel electrophoresis (2-DE) coupled with mass spectrometry to explore the cellular targets of MED and to unravel the molecular mechanisms underlying the antitumor activity of MED in HeLa cells. Our proteomic data showed that triosephosphate isomerase (TPI) and 6-phosphogluconolactonase (PGLS), which participate in glycolysis and the PPP, respectively, were significantly downregulated by MED treatment. Functional studies revealed that the expression levels of several other enzymes involved in glycolysis and the PPP, including hexokinase 2 (HK2), phosphofructokinase 1 (PFKM), aldolase A (ALDOA), enolase 1 (ENO1), lactate dehydrogenase A (LDHA), and glucose-6-phosphate dehydrogenase (G6PD), were also reduced in a dose-dependent manner. Moreover, the LDHA and G6PD enzymatic activities in HeLa cells were inhibited by MED, and overexpression of these downregulated enzymes rescued HeLa cells from the growth inhibition induced by MED. Our data suggest that MED suppresses HeLa cell growth by inhibiting glycolysis and the PPP, which provides a mechanistic basis for the development of new therapeutics against cervical cancer.

Targeting BRK-Positive Breast Cancers with Small-Molecule Kinase Inhibitors.

Approximately 80% of breast cancers overexpress the kinase breast tumor kinase (BRK)/protein tyrosine kinase 6, which has various oncogenic roles in breast cancer cell proliferation, survival, and migration. However, BRK inhibitors have yet to be explored as possible therapeutic tools. In this study, we used a parallel compound-centric approach to discover a new class of pharmaceutical agents, exemplified by XMU-MP-2, as potent and selective BRK inhibitors. XMU-MP-2 exhibited target-specific inhibition of BRK kinase activity and disrupted signaling pathways mediated by this activity, thereby reducing proliferation in BRK-positive breast cancer cells. In mouse xenograft models, XMU-MP-2 repressed the growth of tumors driven by oncogenic BRK, including BRK-transformed Ba/F3 cells and BRK-positive breast cancer cells. Notably, XMU-MP-2 cooperated strongly with HER2 inhibitor or ER blockade to block breast cancer cell proliferation in vitro and in vivo Overall, our findings offer a preclinical proof of concept for therapeutic targeting of the BRK kinase in breast cancer. Cancer Res; 77(1); 175-86. ©2016 AACR.

Structure Determinants of Lagunamide A for Anticancer Activity and Its Molecular Mechanism of Mitochondrial Apoptosis.

Marine natural products are served as attractive source of anticancer therapeutics, with the great success of "first-in-class" drugs, such as Yondelis, Halaven, and Brentuximab vendotin. Lagunamides A-C from marine cyanobacterium, Lyngbya majuscula, exhibit exquisite growth inhibitory activities against cancer cells. In this study, we have systematically investigated the structure-activity relationships (SARs) of a concise collection of lagunamide A and its analogues constructed by total chemical synthesis against a broad panel of cancer cells derived from various tissues or organs, including A549, HeLa, U2OS, HepG2, BEL-7404, BGC-823, HCT116, MCF-7, HL-60, and A375. The R configuration of lagunamide A at C-39 position was found to be the structure determinant for anticancer activity. Further molecular mechanism study in A549 cells revealed that lagunamide A induced caspase-mediated mitochondrial apoptosis. Accompanied with the dissipation of mitochondrial membrane potential (Δφm) and overproduction of reactive oxygen species (ROS), lagunamide A led to mitochondrial dysfunction and finally caused cell death. Moreover, both anti- and pro-apoptotic B-cell lymphoma 2 (Bcl-2) family proteins participated in lagunamide A-induced mitochondrial apoptosis, especially myeloid cell leukemia-1 (Mcl-1). Overexpression of Mcl-1 partly rescued A549 cells from lagunamide A-induced apoptosis. This study suggests that lagunamide A may exert anticancer property through mitochondrial apoptosis. Together, our findings would provide insightful information for the design of new anticancer drugs derived from lagunamides.

Pharmacological targeting of kinases MST1 and MST2 augments tissue repair and regeneration.

Tissue repair and regenerative medicine address the important medical needs to replace damaged tissue with functional tissue. Most regenerative medicine strategies have focused on delivering biomaterials and cells, yet there is the untapped potential for drug-induced regeneration with good specificity and safety profiles. The Hippo pathway is a key regulator of organ size and regeneration by inhibiting cell proliferation and promoting apoptosis. Kinases MST1 and MST2 (MST1/2), the mammalian Hippo orthologs, are central components of this pathway and are, therefore, strong target candidates for pharmacologically induced tissue regeneration. We report the discovery of a reversible and selective MST1/2 inhibitor, 4-((5,10-dimethyl-6-oxo-6,10-dihydro-5H-pyrimido[5,4-b]thieno[3,2-e][1,4]diazepin-2-yl)amino)benzenesulfonamide (XMU-MP-1), using an enzyme-linked immunosorbent assay-based high-throughput biochemical assay. The cocrystal structure and the structure-activity relationship confirmed that XMU-MP-1 is on-target to MST1/2. XMU-MP-1 blocked MST1/2 kinase activities, thereby activating the downstream effector Yes-associated protein and promoting cell growth. XMU-MP-1 displayed excellent in vivo pharmacokinetics and was able to augment mouse intestinal repair, as well as liver repair and regeneration, in both acute and chronic liver injury mouse models at a dose of 1 to 3 mg/kg via intraperitoneal injection. XMU-MP-1 treatment exhibited substantially greater repopulation rate of human hepatocytes in the Fah-deficient mouse model than in the vehicle-treated control, indicating that XMU-MP-1 treatment might facilitate human liver regeneration. Thus, the pharmacological modulation of MST1/2 kinase activities provides a novel approach to potentiate tissue repair and regeneration, with XMU-MP-1 as the first lead for the development of targeted regenerative therapeutics.

[Determination of physiological indices in early kernel at different ear position in maize (Zea mays L.) with UV-visible spectrophotometry].

To examine the difference between early kernels at different ear position in maize and the effect of nitrogen supply rates on kernel development, physiological indices in apical and mid-basal kernel 5-20 days after pollination in maize under different nitrogen supply rates (0, 120, 180 and 240 kg x ha(-1)) were determined with UV-Vis spectrophotometry. The results showed that nitrogen supply obviously increased the contents of soluble sugar, sucrose and starch, and the activities of enzymes related to sucrose inversion and starch synthesis. Twenty days after pollination, the kernel volume, dry weight and the contents of soluble sugar, sucrose and starch in apical kernel under nitrogen supply rate of 180 kg x ha(-1) were higher than those with other treatments. Five to twenty days after pollination, higher activities of acid sucrose invertase (AI), neutral sucrose invertase (NI), sucrose synthase (SS), ADPGase and starch synthase in apical kernel were also obtained under nitrogen supply rate of 180 kg x ha(-1), indicating that the sucrose utilization and starch synthesis were improved, the kernel development was promoted and the kernel abortion was reduced, which resulted in higher yield.

Clinical observation on 930 child epilepsy cases treated with anti-epilepsy capsules.

1090 cases of child epilepsy were divided randomly into two groups: the treatment group (930 cases treated with anti-epilepsy capsules) and the control group (160 cases treated with luminal). The results showed that in the treatment group, 534 cases were markedly effective, 241 effective, 96 improved, 46 ineffective, and 13 aggravated, with a total effective rate of 83.33%; while in the control group, 64 cases were markedly effective, 19 effective, 38 improved, 29 ineffective, and 10 aggravated, with a total effective rate of 51.88%. The treatment group showed an obviously higher total effective rate than that in the control group (P < 0.01). After treatment, cases in the two groups all had lower frequency of epilepsy attacks and shorter duration of each attack as compared with that before treatment (P < 0.01), but the situation was obviously better in the treatment group. The anti-epilepsy capsules had very good effect on various types of epilepsy, especially on autonomic epilepsy and on epilepsies due to wind, phlegm, or terror as differentiated in TCM. After treatment, the recovery rate shown by EEG examination was 54.3% in the treatment group, while 38.4% in the control group, the former being obviously higher than the latter (P < 0.01).