Search

Chinese sprangletop [Leptochloa chinensis (L.) Nees] is one grass weed severely affecting rice (Oryza sativa L.) growth in paddies in China. Cyhalofop-butyl is the main herbicide used to control grass weeds in Chinese paddy fields, especially for controlling L. chinensis; however, L. chinensis has evolved resistance to cyhalofop-butyl due to continuous and extensive application. To investigate cyhalofop-butyl resistance levels and mechanisms in L. chinensis in some of the Chinese rice areas, 66 field populations were collected and treated with cyhalofop-butyl. Of these tested populations, 10 showed a high level of resistance to cyhalofop-butyl; the 50% effective dose ranged within 108.4 to 1,443.5 g ai ha−1 with resistance index values of 9.1 to 121.8 when compared with the susceptible population. Acetyl-coenzyme A carboxylase genes (ACCase) of susceptible and all 10 resistant populations were amplified and sequenced. Among them, Ile-1781-Leu, Trp-2027-Cys, Trp-2027-Ser, and Ile-2041-Asn mutations were found in five resistant populations. No known resistance-related mutations were found in the other five resistant populations, indicating that resistance to cyhalofop-butyl in these populations was likely to be endowed by non–target site resistance mechanisms. Notably, the Ile-1781-Leu and Trp-2027-Cys substitutions have previously been reported, but this is the first report of Trp-2027-Ser and Ile-2041-Asn mutations in L. chinensis. Furthermore, three derived cleaved amplified polymorphic sequence methods were developed to rapidly detect these mutations in L. chinensis.

Colorectal cancer occurs throughout the world but is most common in developed countries. Cancer progression is believed to be driven by genetic mutations in this complex condition. Risk factors for developing colorectal cancer include a genetic family history, long-term ulcerative colitis, and colonic polyps. The use of baicalin has been reported to be clinically efficacious against colon tumors in Asian countries despite an unclear mechanism of action. Several cancers have been found to be biologically dependent on the specificity protein 1 (sp1) transcription factor family. We hypothesized that baicalin may exert its chemotherapeutic effects by sp1 downregulation. Using the SW480 human colorectal cancer cell line, we investigated the physiological properties of baicalin. Our experiments were designed toward clarifying three goals: (a) to determine the mRNA expression profile of transcription factors in colorectal cancer patients using a microarray-based analysis; (b) to determine the effects of baicalin on the sp1 transcription factor with western blotting and reporter cell assays; and (c) to contrast the effects of mithramycin-A (an sp1 transcription factor inhibitor) and baicalin using western blotting and reporter cell assays. Both baicalin and mithramycin-A downregulated sp1 expression, attenuated SW480 cell proliferation, and increased cell apoptosis. Baicalin inhibited sp1 expression and led to SW480 apoptosis, thus clarifying the effect of this traditional Chinese medicine compound in the treatment of colon cancer.

Additional file 5: Fig. S5. sEV miRNA expression profile. (A–C) sEVs were isolated from glutamate (15 μM), GABA (500 μM), or PBS-treated neurons, and the miRNA composition of sEVs was compared by miRNA-sequencing. (A) miRNA sequencing showed 41 upregulated miRNAs in GABA sEVs and 14 downregulated miRNAs in glutamate sEVs compared to Ctrl sEVs. 4 repeated miRNAs were identified in both groups. (B) There were 64 downregulated miRNAs in GABA sEVs and 47 upregulated miRNAs in glutamate sEVs compared with Ctrl sEVs. A total of 12 miRNAs overlapped in both groups. (C) An aggregate of 16 miRNAs (p < 0.05).

Additional file 7: Fig. S7. sEVs with inhibited or over-expressed miR-132 abrogated Glutamate/GABA sEVs induced alterations of apoptotic molecules and synapse-related proteins in APP/PS1 mice, respectively. (A) Cy3-labeled agomir-132 was loaded into sEVs and injected into mice. Twenty-four hours later, the brain slices showed overlapped Cy3-labeled agomir-132-loaded sEVs (red) and NeuN+-neurons (green). (B-O) The mice were injected with glutamate sEVs/Ctrl sEVs and agomir-132/agomir ctrl (B-H) and GABA sEVs/Ctrl sEVs and antagomir-132/antagomir ctrl (I-O)-loaded sEVs at every other day for seven consecutive injections. The expression of apoptotic molecules (cleaved Caspase-3, Bax, and Bcl-2) and synapse-related proteins (Synapsin I, Synapsin I (phospho S9), postsynaptic density-95 (PSD-95)) in the brain lysate of mice was examined by Western blot. n = 3. Data are presented as the mean ± SEM, *p

Additional file 4: Fig. S4. Internalization of PKH-26 labeled sEVs by neurons in APP/PS1 mice. (A) sEVs were labeled with PKH26 (red) and injected via the tail vein in mice. Brain slices were harvested and subjected to the immunostaining of the neuron marker NeuN (green) and visualized. The control group mice were injected with PKH26 only without sEVs. n = 3.

Additional file 1: Fig. S1. sEVs-depleted medium had no effect on the cell viability of Aβ-treated neurons. (A) Image of cultured primary cortical neurons on DIV 15. (B) Immunostaining of cortical cultures on DIV 15 for the detection of GFAP+ glial cells (red) vs. NeuN+ neurons (green). (C) The morphology of sEVs from glutamate or GABA-treated neurons was observed under a transmission electron microscope. Glutamate sEVs = sEVs derived from 10 μM glutamate-treated neurons, GABA sEVs = sEVs derived from 300 μM GABA-treated neurons. (D-F) Neurons were incubated with PBS (the same amount as the solvent of the sEVs), sEVs and sEV-depleted medium (Negative control, NC), then with Aβ as before. Neuronal viability in Ctrl-sEV-treated group (D), glutamate-sEV-treated group (E), GABA-sEV-treated group (F) and NC group were compared using CCK-8. n = 3. Data are presented as the mean ± SEM, ***p

Additional file 3: Fig. S3. Function of sEVs from neurons treated by various concentrations of glutamate or GABA. (A) Primary neurons on DIV 15 were treated with a gradient concentration of Aβ1–42 for 48 h. Cell viability was measured using a CCK-8 assay. (B–I) sEVs were isolated from low concentrations of glutamate (5 μM)/GABA (100 μM), or high concentrations of glutamate (15 μM)/GABA (500 μM) and added to the culture media. Cell viability was measured using CCK-8 (D, E, H, I) and TUNEL assays (B, C, F, G). Glutamate5 sEVs = sEVs derived from 5 μM glutamate treated neurons, GABA100 sEVs = sEVs derived from 100 μM GABA-treated neurons, Glutamate15 sEVs = sEVs derived from 15 μM glutamate-treated neurons, GABA500 sEVs = sEVs derived from 500 μM GABA treated neurons, Ctrl sEVs = sEVs derived from PBS-treated neurons. n = 3. Data are presented as the mean ± SEM, *p

Additional file 6: Fig. S6. sEVs with inhibited or over-expressed miR-132 abrogated Glutamate/GABA sEVs induced alterations of apoptotic molecules in Aβ treated neurons, respectively. (A, B) TUNEL- positive rate of neurons in Fig. 6C and D were calculated. (C–J) Neurons were cotreated with glutamate sEVs and miR-132 mimic-loaded sEVs (C, E–G) or GABA sEVs and miR-132 inhibitor-loaded sEVs (D, H–J). 24 h later, neurons were subjected to Aβ treatment for another 48 h. Then, the expression of apoptotic molecules (cleaved Caspase-3, Bax, and Bcl-2). (K, L) The proportion of TUNEL-positive cells in brain slices in Fig. 6E and F was detected. n = 3. Data are presented as the mean ± SEM, *p

Additional file 2: Fig. S2. Internalization of PKH26 labeled sEVs in primary cultured neurons. (A) sEVs were labeled with PKH26 (red) and added to primary neurons. A control group (only PKH26 without sEVs) was established to exclude false-positive staining caused by the dye. Then, the neurons were immunostained with βIII-tubulin (green) and visualized under a confocal microscope. PKH26 -labeled sEVs were indicated by white arrows. n = 3.

Background. Cerebral ischemic stroke is one of the severe diseases with a pathological condition that leads to nerve cell dysfunction with seldom available therapy options. Currently, there are few proven effective treatments available for improving cerebral ischemic stroke outcome. However, recently, there is increasing evidence that inhibition of histone deacetylase (HDAC) activity exerts a strong protective effect in in vivo and vitro models of ischemic stroke. Review Summary. HDAC is a posttranslational modification that is negatively regulated by histone acetyltransferase (HATS) and histone deacetylase. Based on function and DNA sequence similarity, histone deacetylases (HDACs) are organized into four different subclasses (I-IV). Modifications of histones play a crucial role in cerebral ischemic affair development after translation by modulating disrupted acetylation homeostasis. HDAC inhibitors (HDACi) mainly exert neuroprotective effects by enhancing histone and nonhistone acetylation levels and enhancing gene expression and protein modification functions. This article reviews HDAC and its inhibitors, hoping to find meaningful therapeutic targets. Conclusions. HDAC may be a new biological target for cerebral ischemic stroke. Future drug development targeting HDAC may make it a potentially effective anticerebral ischemic stroke drug.