A mitochondria-targeted fluorescent sensor for imaging endogenous peroxynitrite changes in acute lung injury.

Acute lung injury (ALI) is a serious pulmonary inflammatory disease resulting from excessive reactive oxygen species (ROS) which could cause the damage of the alveolar epithelial cells and capillary endothelial cells. Peroxynitrite, as one of short-lived reactive oxygen species, is closely related to the process of ALI. Thus, it is important to monitor the fluctuation of peroxynitrite in living system for understanding the process of ALI. Herein, the novel mitochondria-targeted fluorescent probe BHMT was designed to respond to peroxynitrite and pH with distinct fluorescence properties respectively. The absorption spectrum of the probe BHMT exhibited a notable red shift as the pH value declined from 8.8 to 2.6. Upon reaction with peroxynitrite, BHMT had a significant increase of fluorescence intensity (63-fold) with maintaining a detection limit of only 43.7 nM. Furthermore, BHMT could detect the levels of endogenous peroxynitrite and image the intracellular pH in ratiometric channels utilizing cell imaging. In addition, BHMT was successfully applied to revealing the relationship between the peroxynitrite and the extent of ALI. Thus, these results indicated the probe BHMT could be a potential tool for diagnosing the early stage of ALI and revealed the peroxynitrite was likely to be a crucial therapeutic target in ALI treatment.

FAK inhibitors in cancer, a patent review - an update on progress.

INTRODUCTION: Focal adhesion kinase (FAK) is a cytoplasmic non-receptor tyrosine kinase over-expressed in various malignancies which is related to various cellular functions such as adhesion, metastasis and proliferation. AREAS COVERED: There is growing evidence that FAK is a promising therapeutic target for designing inhibitors by regulating the downstream pathways of FAK. Some potential FAK inhibitors have entered clinical phase research. EXPERT OPINION: FAK could be an effective target in medicinal chemistry research and there were a variety of FAKIs have been patented recently. Here, we updated an overview of design, synthesis and structure-activity relationship of chemotherapeutic FAK inhibitors (FAKIs) from 2017 until now based on our previous work. We hope our efforts can broaden the understanding of FAKIs and provide new ideas and insights for future cancer treatment from medicinal chemistry point of view.

A highly chromogenic selective Rhodamine-chloride-based fluorescence probe activated by cysteine and application in living cells and zebrafish.

Cysteine (Cys), one of the biological thiols, which plays critical roles in biological system regulating the balance of redox homeostasis. In order to monitor the level of Cys in the living cells and organisms, a chromogenic fluorescence probe Rhocl-Cys based on Rhodamine chloride exhibiting the preferable performance of fluorescence turn-on response reacting with Cys was presented. Rhocl-Cys responded rapidly to Cys within 20 min, and had stable fluorescence intensity within pH 6.0-10.0, high selectivity towards Cys and the anti-inference capability with a low detection limit of 0.80 µM. In particular, Rhocl-Cys could qualitatively and quantitatively monitor the level of endogenous and exogenous Cys in living cells and successfully apply to zebrafish detecting Cys. Therefore, these results might further provide the basis exploring the role of Cys in biological system and facilitate as clinical diagnostic molecular tools.

Hydrogen sulfide treatment retrieves the inhibition of growth and development characteristics in silkworm (Bombyx mori) via phosphoacetyl glucosamine mutase gene knock down.

Phosphoacetyl glucosamine mutase (PGM) is the key gene for glycolysis of important metabolic pathways in silkworm, and H2 S (7.5 µM) can promote the growth and development of silkworm. Herein, we used body cavity injection of small-interfering RNA (siRNA) to interfere with the PGM gene in H2 S-treated silkworms. After RNA interference (RNAi), we investigated the growth and development of the silkworm. H2 S treatment could significantly recover the inhibition of body weight, cocoon weight, cocoon shell weight, and cocoon shell ratio by knocking down PGM gene in silkworm, without significant effects on eggs laying and production, and then analyzed the mRNA expression of PGM gene. The interference of siRNA significantly decreased the expression of targeted PGM gene and was concentrated in 48 h followed by gradual recovery. Three interference fragments also showed different interference effects, and siRNA of PGM-3 exerted the highest interference effect to the target gene expression. Fat body had the highest mRNA expression of PGM gene, and the best interference effect was observed after siRNA injection. The results showed that the gene based on H2 S treatment may have an important impact on the growth and development of silkworm by affecting its metabolic pathway.

Transcriptome Analysis Reveals the Gene Expression Changes in the Silkworm (Bombyx mori) in Response to Hydrogen Sulfide Exposure.

Hydrogen sulfide (H2S) has been recognized for its beneficial influence on physiological alterations. The development (body weight) and economic characteristics (cocoon weight, cocoon shell ratio, and cocoon shell weight) of silkworms were increased after continuous 7.5 µM H2S treatment. In the present study, gene expression changes in the fat body of silkworms at the 5th instar larvae in response to the H2S were investigated through comparative transcriptome analysis. Moreover, the expression pattern of significant differentially expressed genes (DEGs) at the 5th instar larvae was confirmed by quantitative real-time PCR (qRT-PCR) after H2S exposure. A total of 1200 (DEGs) was identified, of which 977 DEGs were up-regulated and 223 DEGs were down-regulated. Most of the DEGs were involved in the transport pathway, cellular community, carbohydrate metabolism, and immune-associated signal transduction. The up regulated genes under H2S exposure were involved in endocytosis, glycolysis/gluconeogenesis, the citrate cycle (TCA cycle), and the synthesis of fibroin, while genes related to inflammation were down-regulated, indicating that H2S could promote energy metabolism, the transport pathway, silk synthesis, and inhibit inflammation in the silkworm. In addition, the expression levels of these genes were increased or decreased in a time-dependent manner during the 5th instar larvae. These results provided insight into the effects of H2S on silkworms at the transcriptional level and a substantial foundation for understanding H2S function.

Two birds with one stone: a NIR fluorescent probe for mitochondrial protein imaging and its application in photodynamic therapy.

Mitochondrial proteins, most of which are encoded in the nucleus and the rest of which are regulated by the mitochondrial genome, play pivotal roles in essential cellular functions. However, fluorescent probes that can be used for monitoring mitochondrial proteins have not yet been widely developed, thereby severely limiting the exploration of the functions of proteins in mitochondria. Towards this end, here we propose a near-infrared (NIR) fluorescence probe MPP to effectively illuminate the dynamic changes in mitochondrial proteins in live cells under oxidative stress, with excellent temporal and spatial resolution. Of particular importance, MPP extends the study of the pharmacology involved in apoptosis induced by anti-cancer drugs (hydroxycamptothecin (HCPT), epirubicin (Epi) and cyclophosphamide (CPA)) for the first time. Furthermore, employing a protein-activatable strategy, this probe could serve as an excellent phototherapeutic agent in photodynamic therapy (PDT). Finally, in vivo experiments suggest that this versatile probe can be used to image tumors in HeLa tumor-bearing mice for 24 h, which demonstrates that our probe could play a dual role as a robust phototherapeutic and imaging agent.

Evaluation of the Metabolic Effects of Hydrogen Sulfide on the Development of Bombyx mori (Lepidoptera: Bombycidae), Using Liquid Chromatography-Mass Spectrometry-Based Metabolomics.

Hydrogen sulfide (H2S) is a highly poisonous gas with an unpleasant smell of rotten eggs. Previous studies of H2S have primarily focused on its effects on mammalian nervous and respiratory systems. In this study, silkworm developmental parameters and changes in metabolites in response to H2S exposure were investigated using a hemolymph metabolomic approach, based on liquid chromatography-mass spectrometry (LC-MS). The developmental parameters, body weight, cocoon weight, cocoon shell weight, and cocoon shell ratio, were noticeably increased following H2S exposure, with the greatest effects observed at 7.5-µM H2S. Metabolites upregulated under H2S exposure (7.5 µM) were related to inflammation, and included (6Z, 9Z, 12Z)-octadecatrienoic acid, choline phosphate, and malic acid, while hexadecanoic acid was downregulated. Identified metabolites were involved in biological processes, including pyrimidine, purine, and fatty acid metabolism, which are likely to affect silk gland function. These results demonstrate that H2S is beneficial to silkworm development and alters metabolic pathways related to spinning function and inflammation. The present study provides new information regarding the potential functions of H2S in insects and metabolic pathways related to this phenomenon.

Anticancerous potential of polysaccharides sequentially extracted from Polygonatum cyrtonema Hua in Human cervical cancer Hela cells.

The anticancerous effects of PCHPs (HBSS, CHSS, DASS, and CASS) were investigated on Human cervical cancer Hela cells proliferation inhibition, cytotoxicity, caspase-3 activity, cell cycle, and apoptosis. The inhibition rate was expressed as CASS > HBSS > CHSS > DASS, with the maximum inhibition of 74.453 ± 3.399%. Cell cytotoxicity was observed (CASS > CHSS > HBSS > DASS) with the maximum cell death rate of 82.472 ± 3.488%. The caspase-3 activity was induced by CASS > HBSS > DASS > CHSS, with the maximum multiple of 2.954 ± 0.103. CASS induced cell cycle block at the G2/M phase by elevating mRNA expression of CyclinD1, p21, p53 and Wee1, and lowering the expression of Survivin, CHK2, Wee1, CyclinB1, and CDK-1. CASS enhanced the mRNA expression of DR3, DR5, FasL, FADD, PARP, TNF- α, TNF- R1, TRDAA, caspases-8, caspases-10 and the protein expression of FasL and caspases-8, -10 in the death receptor pathway; while, lowered the mRNA expression of antiapoptotic genes (Bcl - 2 and Bcl-xL) and the protein expression of Bcl - 2. The mRNA expression of apoptosis genes (Bak, Cytc, Puma, and caspases-3, -7, -9) and the protein expression of caspases-3, -9 of mitochondria pathway was up regulated which led to cell apoptosis.

Effects of sulfated, phosphorylated and carboxymethylated modifications on the antioxidant activities in-vitro of polysaccharides sequentially extracted from Amana edulis.

In this study, four sequentially extracted polysaccharides (AEPs) from Amana edulis were modified by sulphation, phosphorylation, and carboxylation modifications (S-AEPs, P-AEPs, C-AEPs), and compared for their anti-oxidant activities. After modification, sugar and protein contents were decreased and uronic acid content was increased in comparison to native AEPs. UV absorption showed similar maximum absorption peaks of modified derivatives which indicated their homogeneous nature. FTIR spectra confirmed the conversion of hydroxyl groups to OS, COO, and POH bonds, respectively. The phosphorylated derivatives (P-AEPs) displayed the highest DPPH, hydroxyl radical, and ferrous ions radical scavenging abilities. Sulfated polysaccharides (S-AEPs) were observed with high reducing ability. The C-AEPs maintained the stable antioxidant properties after carboxylation modification. Our results indicated that the chemical modification of different polysaccharide components has significantly affected their antioxidant potential for their use in food industry and human health.

Transcriptome analysis reveals gene expression changes of the fat body of silkworm (Bombyx mori L.) in response to selenium treatment.

A comparative transcriptome analysis was conducted to investigate the gene expression changes in the fat body of silkworm after treatment with different concentrations (50 µM and 200 µM) of selenium (Se). 912 differential expression genes (DEGs) (371 up-regulated and 541 down-regulated) and 1420 DEGs (1078 up-regulated and 342 down-regulated) were identified in silkworm fat body treated with 50 µM and 200 µM of Se, respectively. In case of 50 µM group, DEGs were mainly enriched in the peroxisome pathway and fatty acid metabolism pathway, and later were associated with antioxidant defense and nutrition regulation. After 200 µM Se-treatment, DEGs were mainly located in the glycerolipid metabolism and arachidonic acid metabolism pathways, which further encoded detoxification related genes. Furthermore, 32 candidate DEGs from these pathways had been selected to confirm the RNA-seq data. Among these DEGs, 14 genes were up-regulated in the 50 µM Se-treated group (only three genes in the 200 µM Se-treated group) which were involved in lipid metabolism and antioxidant defense, and 13 up-regulated genes (only two genes were up-regulated in the 50 µM Se-treated group) were involved in detoxification of the 200 µM Se-treated group. These changes showed that lower concentration of Se could regulate the nutrition and promote antioxidation pathways; whereas, high levels of Se promoted the detoxification of silkworm. These findings can be helpful to understand the possible mechanisms of Se action and detoxification in silkworm and other insects.

Effect of Dietary Selenium Supplementation on Growth and Reproduction of Silkworm Bombyx mori L.

The effects of selenium (Se) on the growth and reproduction of the Lepidoptera insect, the silkworm, Bombyx mori L were investigated. Initially, the silkworms were divided into eight groups (150 larvae/group) on the basis of feeding with mulberry leaves saturated with different concentrations of Se (25, 50, 100, 125, 150, 175, and 200 µM) and control from the first day of the fourth instar larvae. After feeding, growth and reproductive performance of B. mori L. were investigated with standard techniques used in sericulture. After the data analysis, 50 µM of Se was recognized as the optimal level which positively influenced the growth and production, with prolonged stage of larvae, increased larval, cocoon, and pupal weights, and enhanced number of eggs laid by the female moth as compared to the control group. On the contrary, 200 µM of Se treatment displayed toxic to silkworm and induced significant decrease in the growth, cocoon production, and reproduction. The weight of the cocoon shell, the cocoon shell ratio, number of eggs produced, and fertilization ratio in all the Se-treated groups were lower than the control group. The present study indicated that lower levels of (50 µM) of Se can promote the larval and pupal growth of the B. mori L. resulting in the higher yield of cocoon crop and significantly influencing the fecundity, while high concentration was toxic to silkworm. Our data supply the novel application of Se which could be highly beneficial to sericulture farmers.

Liquiritin from Glycyrrhiza uralensis Attenuating Rheumatoid Arthritis via Reducing Inflammation, Suppressing Angiogenesis, and Inhibiting MAPK Signaling Pathway.

Among the various treatments, induction of synoviocyte apoptosis by natural products during a rheumatoid arthritis (RA) pathological condition can be considered to have vast potential. However, it is unclear that liquiritin, a kind of natural flavonoid extracted from the roots of Glycyrrhiza uralensis, induced the apoptosis of the synovial membrane and its molecular mechanism. In this study, interleukin-1ß (IL-1ß)-RA-FLS cells were incubated with different concentrations of liquiritin. An MTT assay, Hoechst 33342 staining, JC-1 staining, and Western blot were used to check the viability, cell apoptosis, mitochondrial membrane potential changes, and the expression of related proteins, respectively. In vivo, a TUNEL assay and HE staining of tissue were used for histopathological evaluation. Our results showed that liquiritin significantly inhibited the proliferation of IL-1ß-induced-RA-FLS, promoted nuclear DNA fragmentation, and changed the mitochondrial membrane potential to accelerate cell apoptosis. Liquiritin downregulated the ratio of Bcl-2/Bax and inhibited the VEGF expression and phosphorylation of JNK and P38. Moreover, liquiritin improved the clinical score of rheumatism, inflammatory infiltration, and angiogenesis and induced apoptosis of the synovial tissue in vivo. Hence, liquiritin ameliorates RA by reducing inflammation, blocking MAPK signaling, and restraining angiogenesis.