Discovery of novel 3-(1H-pyrazol-4-yl)-1H-indazole derivatives as potent type II TRK inhibitors against acquired resistance.

Tropomyosin receptor kinase (TRK) is a promising target for treating NTRK fusion cancers. The solvent front and xDFG mutations induced by larotrectinib and entrectinib result in acquired resistance in advanced-stage patients. In this study, we report a highly potent and selective type II TRK inhibitor, 40l, developed using a structure-based design strategy. Compound 40l significantly suppressed Km-12, Ba/F3-TRKAG595R, and Ba/F3-TRKAG667C cell proliferation. In biochemical and cellular assays, 40l showed better inhibitory activity against TRKAG667C than that by the positive control, selitrectinib. Additionally, it induced apoptosis of Ba/F3-TRKAG595R and Ba/F3-TRKAG667C cells in a dose-dependent manner. Furthermore, 40l showed good selectivity for a panel of 41 kinases. In vitro assays indicated that 40l possessed outstanding plasma stability and moderate liver microsomal stability. Based on the above results, compound 40l could be further optimized to overcome the solvent front and xDFG TRK mutations.

Inhibition Roles of Calcium in Cadmium Uptake and Translocation in Rice: A Review.

Cadmium (Cd) contamination in rice grains is posing a significant threat to global food security. To restrict the transport of Cd in the soil-rice system, an efficient way is to use the ionomics strategy. Since calcium (Ca) and Cd have similar ionic radii, their uptake and translocation may be linked in multiple aspects in rice. However, the underlying antagonistic mechanisms are still not fully understood. Therefore, we first summarized the current knowledge on the physiological and molecular footprints of Cd translocation in plants and then explored the potential antagonistic points between Ca and Cd in rice, including exchange adsorption on roots, plant cell-wall composition, co-transporter gene expression, and transpiration inhibition. This review provides suggestions for Ca/Cd interaction studies on rice and introduces ionomics research as a means of better controlling the accumulation of Cd in plants.

Renal ischemia-reperfusion injury in rabbits evaluated with quantitative susceptibility mapping: an animal study.

Background: There were no effective noninvasive methods to diagnose renal ischemia-reperfusion injury (IRI), which is a major clinical problem. The objective of this study was to explore the feasibility of the quantitative susceptibility mapping (QSM) technique in evaluating the dynamic changes in the renal IRI process. Methods: A total of 36 New Zealand rabbits were randomly assigned to the IRI group (n=30) and the sham group (n=6). All rabbits underwent magnetic resonance imaging (MRI) examination, including T2-weighted imaging and QSM before the operation (pre-IRI) and 1, 12, 24, and 48 h after the operation (IRI-1h, IRI-12h, IRI-24h, and IRI-48h, respectively). Regions of interest were manually delineated in the outer medulla. All specimens were stained with hematoxylin and eosin (HE) and glutathione peroxidase 4 (GPX4). The pathological score of renal injury and the average optical density value of GPX4 were calculated. The repeated measurement analysis of variance (ANOVA) and Spearman correlation analysis were used to compare the differences between the susceptibility values and determine the correlation. Results: In the IRI group, the susceptibility values of the outer medulla at the pre-IRI, IRI-1h, IRI-12h, IRI-24h, and IRI-48h time points were (42.83±7.83)×10-3, (-5.33±6.28)×10-3, (6.50±3.94)×10-3, (12.00±3.74)×10-3, and (22.00±6.81)×10-3 ppm, respectively. The susceptibility values significantly differed among the different time points (P<0.001). The susceptibility values had a negative correlation with the scores of cell edema (ρ=-0.61; P=0.002) and the average optical density value of GPX4 (ρ=-0.70; P<0.001). The susceptibility values had a positive correlation with iron content (ρ=0.79; P<0.001), the scores of cell necrosis (ρ=0.71; P<0.001), interstitial inflammation (ρ=0.60; P=0.002), cast (ρ=0.75; P<0.001), and the total pathological score of renal injury (ρ=0.51; P=0.01). Conclusions: QSM can be used as a noninvasive method to assess the dynamic changes of the outer medulla in the early stage of renal IRI in rabbits.

Design, synthesis and biological evaluation of novel indolin-2-one derivatives as potent second-generation TRKs inhibitors.

Tropomyosin receptor kinases (TRKs) are effective targets for anti-cancer drug discovery. The first-generation type I TRKs inhibitors, larotrectinib and entrectinib, exhibit durable disease control in the clinic. The emergence of acquired resistance mediated by secondary mutations in the TRKs domain significantly reduces the therapeutic efficacy of these two drugs, indicating an unmet clinical need. In this study, we designed a potent and orally bioavailable TRK inhibitor, compound 24b, using a molecular hybridization strategy. Compound 24b exhibited significant inhibitory potency against multiple TRK mutants in both biochemical and cellular assays. Furthermore, compound 24b induced apoptosis of Ba/F3-TRKAG595R and Ba/F3-TRKAG667C cells in a dose-dependent manner. Additionally, compound 24b exhibited moderate kinase selectivity. In vitro stability revealed that compound 24b showed excellent plasma stability (t1/2 > 289.1 min) and moderate liver microsomal stability (t1/2 = 44.3 min). Pharmacokinetic studies have revealed that compound 24b is an orally bioavailable TRK inhibitor with a good oral bioavailability of 116.07%. These results indicate that compound 24b be used as a lead molecule for further modifications to overcome drug-resistant mutants of TRK.

Design, synthesis and biological evaluation of pyrazolo[3,4-b]pyridine derivatives as TRK inhibitors.

Tropomyosin receptor kinases (TRKs) are associated with the proliferation and differentiation of cells, and thus their continuous activation and overexpression cause cancer. Herein, based on scaffold hopping and computer-aid drug design, 38 pyrazolo[3,4-b]pyridine derivatives were synthesised. Further, we evaluated their activities to inhibit TRKA. Among them, compound C03 showed acceptable activity with an IC50 value of 56 nM and it inhibited the proliferation of the Km-12 cell line with an IC50 value of 0.304 µM together with obvious selectivity for the MCF-7 cell line and HUVEC cell line. Furthermore, compound C03 possessed good plasma stability and low inhibitory activity to a panel of cytochrome P450 isoforms except CYP2C9. Overall, C03 has potential for further exploration.

Design, synthesis, and biological evaluation of potent FAK-degrading PROTACs.

FAK mediated tumour cell migration, invasion, survival, proliferation and regulation of tumour stem cells through its kinase-dependent enzymatic functions and kinase-independent scaffolding functions. At present, the development of FAK PROTACs has become one of the hotspots in current pharmaceutical research to solve above problems. Herein, we designed and synthesised a series of FAK-targeting PROTACs consisted of PF-562271 derivative 1 and Pomalidomide. All compounds showed significant in vitro FAK kinase inhibitory activity, the IC50 value of the optimised PROTAC A13 was 26.4 nM. Further, A13 exhibited optimal protein degradation (85% degradation at 10 nM). Meantime, compared with PF-562271, PROTAC A13 exhibited better antiproliferative activity and anti-invasion ability in A549 cells. More, A13 had excellent plasma stability with T1/2 >194.8 min. There are various signs that PROTAC A13 could be useful as expand tool for studying functions of FAK in biological system and as potential therapeutic agents.

[Arbuscular mycorrhizal fungi enhanced cadmium uptake in Photinia frase through altering root transcriptomes and root-associated microbial communities].

As a non-essential metal, cadmium (Cd) pollution poses severe threats to plant growth, environment, and human health. Phytoextraction using nursery stocks prior to their transplantation is a potential useful approach for bioremediation of Cd contaminated soil. A greenhouse pot experiment was performed to investigate the growth, Cd accumulation, profiles of transcriptome as well as root-associated microbiomes of Photinia frase in Cd-added soil, upon inoculation of two types of arbuscular mycorrhizal fungi (AMF) Sieverdingia tortuosa and Funneliformis mosseae. Compared with the control, inoculation of F. mosseae increased Cd concentrations in root, stem and leaf by 57.2%, 44.1% and 71.1%, respectively, contributing to a total Cd content of 182 µg/plant. KEGG pathway analysis revealed that hundreds of genes involved in 'Mitogen-activated protein kinase (MAPK) signaling pathway', 'plant hormone signal transduction', 'biosynthesis of secondary metabolites' and 'glycolysis/gluconeogenesis' were enriched upon inoculation of F. mosseae. The relative abundance of Acidobacteria was increased upon inoculation of S. tortuosa, while Chloroflexi and Patescibacteria were increased upon inoculation of F. mosseae, and the abundance of Glomerales increased from 23.0% to above 70%. Correlation analysis indicated that ethylene-responsive transcription factor, alpha-aminoadipic semialdehyde synthase, isoamylase and agmatine deiminase related genes were negatively associated with the relative abundance of Glomerales operational taxonomic units (OTUs) upon inoculation of F. mosseae. In addition, plant cysteine oxidase, heat shock protein, cinnamoyl-CoA reductase and abscisic acid receptor related genes were positively associated with the relative abundance of Patescibacteria OTUs upon inoculation of F. mosseae. These finding suggested that AMF can enhance P. frase Cd uptake by modulating plant gene expression and altering the structure of the soil microbial community. This study provides a theoretical basis for better understanding the relationship between root-associated microbiomes and root transcriptomes of P. frase, from which a cost-effective and environment-friendly strategy for phytoextraction of Cd in Cd-polluted soil might be developed.

Fruit extracts from Phyllanthus emblica accentuate cadmium tolerance and accumulation in Platycladus orientalis: A new natural chelate for phytoextraction.

A key challenge for phytoextraction is the identification of high efficiency, growth-supporting, and low cost chelating agents. To date, no substance has satisfied all above criteria. This study investigated nine traditional Chinese herbs and found that Phyllanthus emblica fruit (FPE) extract could be utilised as an optimal chelate for the phytoextraction of cadmium (Cd)-contaminated soils. FPE application into soil at a ratio of 0.1% (w/w) significantly increased extractable Cd (by 43%) compared to the control. The success of FPE as a chelating agent was attributed to high quantities of polyphenol compounds (0.76%) and organic acids (9.6%), in particular, gallic acid (7.6%). Furthermore, antioxidative properties (1.4%) and free amino acids in FPE alleviated Cd-induced oxidant toxicity and enhanced plant biomass. FPE promoted 78% higher phytoextraction efficiency in Platycladus orientalis compared to traditional chelating agents (EDTA). Furthermore, 76% of FPE was degraded 90 days after the initial application, and there was no difference in extractable Cd between the treatment and control. FPE has been commercially produced at a lower market price than other biodegradable chelates. As a commercially available and cost-effective chelator, FPE could be utilised to treat Cd-contaminated soils without adverse environmental impacts.

Azolla incorporation under flooding reduces grain cadmium accumulation by decreasing soil redox potential.

Cadmium (Cd) presents severe risks to human health and environments. The present study proposed a green option to reduce bioavailable Cd. Rice pot experiments were conducted under continuous flooding with three treatments (T1: intercropping azolla with rice; T2: incorporating azolla into soil before rice transplantation; CK: no azolla). The results showed that azolla incorporation reduced soluble Cd by 37% compared with the CK treatment, which may be explained by the decreased soil redox potential (Eh) (r = 0.867, P < 0.01). The higher relative abundance of Methylobacter observed in azolla incorporation treatment may account for dissolved organic carbon increase (r = 0.694; P < 0.05), and hence decreased the Cd availability for rice. Azolla incorporation increased the abundance of Nitrospira, indicating the potentially prominent role of nitrogen mineralization in increasing rice yields. Further, lower soluble Cd decreased the expression of OsNramp5, but increased OsHMA3 levels in rice roots, which decreased Cd accumulation in grains. Through these effects, azolla incorporation decreased Cd concentrations in rice grains by 80.3% and increased the production by 13.4%. The negligible amount of Cd absorbed by azolla would not increase the risk of long-term application. Thus, intercropping azolla with early rice and incorporating azolla into soil before late rice transplantation can contribute to safe production at large scales of double rice cultivation.

Health risk assessment of heavy metal pollution in a soil-rice system: a case study in the Jin-Qu Basin of China.

A regional field survey of a total of 109 pairs of soil and rice samples was conducted to evaluate the health risks posed by heavy metals in the Jin-Qu Basin, China. The studied soils are characterized by acid (pH in mean level of 5.5), carbon rich (soil organic matter in mean of 33.6 g kg-1) and mainly contaminated by Cd (42.2% samples exceeded the standard value of 0.3 mg kg-1 (GB15618-2018)). The spatial distributions of Cd, Pb and Zn exhibited similar geographic trends. 34% and 30% of the rice samples containing Cd and Pb exceeded the threshold value of 0.2 mg kg-1 (GB2762-2017), respectively. The risk estimation of dietary intake had a target hazard quotient value of Cd of 0.918 and a hazard index value for rice consumption of 2.141. Totally, Cd and Pb were found to be the main components contributing to the potential health risks posed by non-carcinogenic effects for local inhabitants.

Salicylic Acid Signals Plant Defence against Cadmium Toxicity.

Salicylic acid (SA), as an enigmatic signalling molecule in plants, has been intensively studied to elucidate its role in defence against biotic and abiotic stresses. This review focuses on recent research on the role of the SA signalling pathway in regulating cadmium (Cd) tolerance in plants under various SA exposure methods, including pre-soaking, hydroponic exposure, and spraying. Pretreatment with appropriate levels of SA showed a mitigating effect on Cd damage, whereas an excessive dose of exogenous SA aggravated the toxic effects of Cd. SA signalling mechanisms are mainly associated with modification of reactive oxygen species (ROS) levels in plant tissues. Then, ROS, as second messengers, regulate a series of physiological and genetic adaptive responses, including remodelling cell wall construction, balancing the uptake of Cd and other ions, refining the antioxidant defence system, and regulating photosynthesis, glutathione synthesis and senescence. These findings together elucidate the expanding role of SA in phytotoxicology.

Endogenous salicylic acid is required for promoting cadmium tolerance of Arabidopsis by modulating glutathione metabolisms.

A few studies with NahG transgenic lines of Arabidopsis show that depletion of SA enhances cadmium (Cd) tolerance. However, it remains some uncertainties that the defence signaling may be a result of catechol accumulation in NahG transgenic lines but not SA deficiency. Here, we conducted a set of hydroponic assays with another SA-deficient mutant sid2 to examine the endogenous roles of SA in Cd tolerance, especially focusing on the glutathione (GSH) cycling. Our results showed that reduced SA resulted in negative effects on Cd tolerance, including decreased Fe uptake and chlorophyll concentration, aggravation of oxidative damage and growth inhibition. Cd exposure significantly increased SA concentration in wild-type leaves, but did not affect it in sid2 mutants. Depletion of SA did not disturb the Cd uptake in either roots or shoots. The reduced Cd tolerance in sid2 mutants is due to the lowered GSH status, which is associated with the decreased expression of serine acetyltransferase along with a decline in contents of non-protein thiols, phytochelatins, and the lowered transcription and activities of glutathione reductase1 (GR1) which reduced GSH regeneration. Finally, the possible mode of SA signaling through the GR/GSH pathway during Cd exposure is discussed.

Exon 47 skipping of fibrillin-1 leads preferentially to cardiovascular defects in patients with thoracic aortic aneurysms and dissections.

Excessive activation of the transforming growth factor beta signaling pathway and disorganized cellular skeleton caused by genetic mutations are known to be responsible for the inherited thoracic aortic aneurysms and dissections (TAAD), a life-threatening vascular disease. To investigate the genotype-phenotype correlation, we screened genetic mutations of fibrillin-1 (FBN1), transforming growth factor-ß receptor-1 (TGFBR1) and transforming growth factor-ß receptor-2 (TGFBR2) for TAAD in 7 affected families and 22 sporadic patients. Of 19 potential mutations identified in FBN1, 11 appeared novel while the others were recurrent. Two mutations were detected in TGFBR2. Eight patients carried no mutation in either of these genes. Characterization of FBN1 c.5917+6T>C in transfected HEK293 cells demonstrated that it caused skipping of exon 47, leading to the loss of the 33th calcium binding epidermal growth factor-like domain associated with Marfan syndrome. Compared with exon 46, skipping of 47 did not cause patients ectopia lentis in all carriers. To correlate genotypes with phenotypes in different human ancestries, we reviewed the published mutational studies on FBN1 and found that the probability of cardiovascular defects were significantly increased in Chinese patients with premature termination codon or splicing mutations than those with missense mutations (91.7 % vs 54.2 %, P = 0.0307) or with noncysteine-involved point mutations than those with cysteine-involved mutations (88.9 % vs 33.3 %, P = 0.0131). Thus, we conclude that exon 47 skipping of FBN1 leads preferentially to cardiovascular defects and human ancestries influence genotype-phenotype correlation in TAAD.

[Risk factors of cognitive impairment after off-pump coronary artery bypass grafting].

OBJECTIVE: To analyze the risk factors of cognitive impairment after off-pump coronary artery bypass grafting (OPCABG). METHODS: A total of 102 patients [male: 82, age: (65.7 ± 7.1) years] undergoing OPCABG in our hospital between January 2009 and December 2010 were divided into postoperative cognitive dysfunction (POCD) group and non-POCD group by the MMSE questionnaire survey conducted at 7 days pre- and post-operation respectively. RESULTS: The incidence of POCD was 48.0% (49/102). Multivariate logistic stepwise regression analysis showed: advanced age (OR = 1.32, 95%CI: 1.10 - 1.46, P = 0.002), smoking (OR = 1.26, 95%CI: 1.18 - 1.32, P = 0.001), hypertension (OR = 1.66, 95%CI: 1.36 - 1.78, P = 0.023), diabetes (OR = 1.62, 95%CI: 1.02 - 2.84, P = 0.032), stroke (OR = 3.32, 95%CI: 1.68 - 6.49, P < 0.001), mitral regurgitation (OR = 1.48, 95%CI: 1.26 - 1.89, P < 0.001), and time of wall clamp (OR = 4.84, 95%CI: 1.08 - 7.28, P < 0.001) were independent risk factors of POCD. CONCLUSION: Advanced age, smoking, hypertension, diabetes, stroke, mitral regurgitation, and prolonged time of wall clamp are major risk factors for POCD in patients undergoing OPCABG.

The efficacy and safety of melatonin in concurrent chemotherapy or radiotherapy for solid tumors: a meta-analysis of randomized controlled trials.

BACKGROUND: Recently, melatonin has been associated with cancer both in vitro and in vivo. However, the value of melatonin in the treatment of cancer remains disputable. Hence, we performed a systematic review of randomized controlled trials (RCTs) of melatonin in solid tumor cancer patients and observed its effect on tumor remission, 1-year survival, and side effects due to radiochemotherapy. METHODS: An electronic search was conducted using the databases Pubmed, Medline, EMBASE, Cochrane library, and CNKI, from inception to November 2011. Trials using melatonin as adjunct treatment concurrent with chemotherapy or radiotherapy for cancer were included. Pooled relative risk (RR) for the tumor remission, 1-year survival, and radiochemotherapy-related side effects were calculated using the software Revman 5.0. RESULTS: The search strategy identified 8 eligible RCTs (n = 761), all of which studied solid tumor cancers. The dosage of melatonin used in the 8 included RCTs was 20 mg orally, once a day. Melatonin significantly improved the complete and partial remission (16.5 vs. 32.6%; RR = 1.95, 95% CI, 1.49-2.54; P < 0.00001) as well as 1-year survival rate (28.4 vs. 52.2%; RR = 1.90; 95% CI, 1.28-2.83; P = 0.001), and dramatically decreased radiochemotherapy-related side effects including thrombocytopenia (19.7 vs. 2.2%; RR = 0.13; 95% CI, 0.06-0.28; P < 0.00001), neurotoxicity (15.2 vs. 2.5%; RR = 0.19; 95% CI, 0.09-0.40; P < 0.0001), and fatigue (49.1 vs. 17.2%; RR = 0.37; 95% CI, 0.28-0.48; P < 0.00001). Effects were consistent across different types of cancer. No severe adverse events were reported. CONCLUSIONS: Melatonin as an adjuvant therapy for cancer led to substantial improvements in tumor remission, 1-year survival, and alleviation of radiochemotherapy-related side effects.

Cadmium stabilization with nursery stocks through transplantation: a new approach to phytoremediation.

Disposal of heavy metal contaminated biomass after phytoremediation is still unfeasible. This paper presents a viable phyto-extraction approach in which metals in contaminated soils are stabilized by nursery stocks before transplantation for greening. In this respect, two pot-experiments are reported comparing seven nursery stocks species exposed to different Cd levels. The first experiment revealed that Cd was mainly stabilized in the roots of all species studied. Greater amounts of Cd were accumulated in the epidermis than cortex plus stele. Cupressus Blue Ice showed greatest tolerance to the 100 and 200 mg kg(-1) Cd stresses. The second experiment additionally evaluated the possible risk of Cd release after transplanting the Cd treated plants into uncontaminated soil. After 120 days of transplantation, the relatively trace amounts of Cd in the roots of Euonymus japonicus, Pittosporum tobira and C. Blue Ice had either been partially transferred into the shoots or released into the soil. The highest Cd concentration increase in bulk soil (0.428 mg kg(-1)), however, was much lower than the environmental quality standard for soils of China (1 mg kg(-1)). The potential effectiveness of this technique in the use of Cd-contaminated soil and further investigation needed in the field trials were also evaluated.

[Effects of open heart surgery under normothermic and hypothermic cardiopulmonary bypass on cytokines and complements].

OBJECTIVE: To explore the detrimental influence of normothermic and hypothermic cardiopulmonary bypass during open heart surgery on cytokines and complements. METHOD: Forty patients with congenital or rheumatic heart disease were randomized into 2 groups to receive normothermic cardiopulmonary bypass (CPB, study group, n=20) or hypothermic CPB (control group, n=20). Venous blood samples were respectively collected at scheduled time points preoperatively, at the end of CPB, and 1,4,7,14 d postoperatively to examine the level of interleukin (IL)-2, tumor necrosis factor (TNF)-alpha, C3, and C4. RESULTS: IL-2 in both groups decreased significantly at the end of CPB, postoperative day 1 and 4, but recovered the normal level at day 7 postoperatively. IL-2 in control group was significantly lower than that in the study group at each time points at the end of CPB and day 1 and 4 postoperatively. TNF-alpha in two groups was both elevated at the time points cited above, and in the study group, recovery of normal TNF-alpha level occurred at day 7 postoperatively, whereas in the control group, the recovery was not achieved until postoperative day 14. C3 in the study group was significantly lower at the time points of the end of CPB, day 1, 7 postoperatively than that in control group, but both elevated above normal at the end of CPB, day 1, and 4 postoperatively; in the study group, C3 became normal at day 7 postoperatively, which occurred in the control group only till day 14 postoperatively. At the end of CPB and day 1 postoperatively, C4 was significantly lower in the study group than in the control group, both below the level measured preoperatively at the time points of the end of CPB, day 1 and 4 postoperatively. CONCLUSION: Open-heart surgery under normothermic CPB has less detrimental influence on cytokines and complements than the operation under hypothermic CPB for better recovery of the patient.