Hovenia dulcis Thunb. is a traditional hepatoprotective Chinese medicine, and in research, much effort has been focused on the protection against alcoholic liver injury. In this study, the protective effects of a fruit ethanol extract of Hovenia dulcis (FE) against APAP-induced acute hepatotoxicity in mice and the possibly involved molecular mechanisms were investigated. Hepatoprotective activity of FE is clearly indicated by histopathological and biochemical examination. Treatment with FE resulted in inhibition of CYP2E1 activity involved in the transformation of APAP in vivo. Expressions of the altered bile acid metabolism and transport-related genes and relative proteins of apoptosis were normalized by preconditioning with FE before APAP treatment. These results suggested FE to alleviate APAP-induced liver injury in a dose-dependent manner by inhibition of cytochrome P450 activity, hepatocyte apoptosis and regulation of bile acid homeostasis imbalance.
Background: YAP/TAZ have been reported to be highly expressed in malignant tumors. This research was aimed to study the expression of TAZ/YAP protein in malignant melanoma and to investigate its relationship to the clinicopathology and prognosis of patients with malignant melanoma. Methods: A total of 156 pairs of malignant melanoma tumor tissues and melanocytic nevi tissues were collected. The expression of TAZ and YAP proteins were detected by immunohistochemistry and western blotting. Relationship between TAZ/YAP and clinical data of malignant melanoma were analyzed. Results: The number of positive cells of TAZ and YAP in malignant melanoma tumor tissues was significantly higher than that in melanocytic nevi tissues, and the expression of both proteins was positively correlated with malignant melanoma (r = 0.687 P<0.01); TAZ and YAP protein expression in malignant melanoma tissue was significantly correlated with tumor thickness, invasive depth grade, lymph node metastasis and TNM stages (P <0.05), and increased with TNM staging. The expression of TAZ and YAP in patients with lymph node metastasis was significantly higher than that in non-metastatic patients (P <0.05). High expression of TAZ/YAP significantly reduced the postoperative survival rate in patients with malignant melanoma. Conclusion: TAZ/YAP is highly expressed in malignant melanoma tumor tissues. The high expression of TAZ/YAP promotes the progression of malignant melanoma and affects the postoperative survival of patients.
Aim: The aim of this study was to further elucidate the mechanism of pterostilbene against UVA/UVB irradiation and the Nuclear factor E2-related factor 2 (Nrf2) signal pathway. Methods: A photo-damage model with UVA/UVB irradiation in HaCat cells was established and used in this study. The dose of pterostilbene was selected through MTS assay. Cell proliferation and apoptosis in Nrf2 and knockdown Nrf2 cells was detected by MTS assay. Expression of CAT, HO-1, and SOD in Nrf2 and knockdown Nrf2 cells was explored by qPCR. Western blot was used to analysis of Nrf2 nuclear translocation changes in Nrf2 and knockdown Nrf2 cells. Protein carbonyl content and MDA content was tested. Results: Our photo-damage model was successfully established and 20J/cm(2) UVA and 57mJ/cm(2) UVB irradiation was the suitable dose for HaCaT cell damage study. UVA/UVB irradiation would affect Nrf2 protein location, especial for 9.75 mu M pterostilbene dose. In addition, cell proliferation could be significantly inhibited by UVA/UVB treatments (P<0.05), whereas, 9.75 mu M pterostilbene treatment can alleviate the photo-damage. UVA/UVB irradiation would lead to decreased expressions of CAT, HO-1, and SOD. Carbonyl content and MDA was significantly changed by UVA/UVB treatments (P<0.05). The adverse events could be reversed by adding 9.75 mu M pterostilbene. Western blot analysis showed that Nrf2 cytoplasm content in UVA/UVB treated cells was reduced and Nrf2 nuclear content was increased, which are different with the normal HaCaT cells without knockdown Nrf2 treatment (P<0.05). The results of cell proliferation, apoptosis, and cell antioxidant capacity in knockdown Nrf2 treated HaCaT cells were also significantly different with the normal HaCaT cells without knockdown Nrf2 treatment (P<0.05). Conclusion: We hypothesize that pterostilbene could play an anti-oxidation role via the Nrf2 signal pathway.
Objective: The study observed the efficacy and safety of dexmedetomidine, propofol and etomidate on brain functional areas in patients undergoing wake-up brain surgery under the guidance of entropy index. Methods: Sixty patients undergoing wake-up brain surgery on brain functional areas were enrolled, and randomly divided into three groups: dexmedetomidine group (group D), propofol group (group P) and etomidate group (group E), 20 in each. The vital signs, entropy indices of each time point, wake-up time, wake-up quality and adverse reaction in the wake-up period were observed and compared. Results: There were no differences in the duration of wake-up, duration of anesthesia, duration of surgery and postoperative wake-up time between the three groups (P> 0.05). The wake-up quality in group D was significantly better than group P and group E (P <0.05), group P was better than group E (P <0.05). The incidence of adverse events in group D was lower than that in groups P and E (P <0.05). The incidence of adverse events in group P was lower than that in group E (P <0.05). Conclusion: Under the guidance of entropy index, anesthesia induced by dexmedetomidine, propofol and etomidate combined with remifentanil can safely and effectively be used to the wake-up brain functional areas surgery, but the wake-up quality with use of dexmedetomidine is highest, and the incidence of adverse events is the lowest during wake-up period.
Levetiracetam (LEV) is an anti-epileptic drug with demonstrated efficacy against generalized seizures. Recent studies have found that LEV affects the release of neurotransmitters by binding to synaptic vesicle protein 2A (SV2A). However, the details of LEV regulation of synaptic transmission remain poorly understood. Here, we used the whole-cell patch-clamp technique to further characterize the effects of LEV on synaptic transmission at a large mammalian central synapse, the calyx of Held. Our results showed that two common forms of vesicle endocytosis, including slow and rapid endocytosis, were dramatically inhibited when slices were incubated in 100 mu M LEV for 1 h, however, the action potential (AP), calcium influx and exocytosis were not affected. Furthermore, by measuring the level of steady-state depression induced by 100 Hz stimulus trains, we found that the steady state level of depression was significantly stronger after LEV treatment, indicating that LEV enhanced short-term depression (STD). Thus, these findings suggested that the mechanisms of the antiepileptic of LEV seem to be mediated, at least partly, by regulating endocytosis and STD.
EAI045 represents a fourth generation allosteric EGFR TKI compound which targets T790M and C797S EGFR mutants. The study reported herein describes a method to explore the distribution of EAI045 in rat tissues as well as to quantify it in plasma. The method used here is an ultra-performance liquid chromatography-tandem mass spectrometry with high sensitivity and selectivity. An ACQUITY UPLC BEN HILIC column with dimensions of 2.1 x 100 mm, 1.7 mu m was used to separate the analytes and IS. As mobile phase acetonitrile as well as 0.1 % of formic acid/water was used combined with an elution gradient and 0.40 mL/min flow rate. This eluent was also used for electrospray ionization in positive ion mode. A mode on multiple reactions monitoring (MRM) was also employed in the quantification. This quantification included the use of targeted segment ions with m/z 384.1 -> 100.8 for EAI045, and m/z 285.1 -> 193.3 for IS, respectively. It was found that the linearity of this method was appropriate and the concentration range could be kept within a range of 2-2000 ng/mL for EAI045 in rat plasma and tissues. The level of EAI045 was found to be highest in the liver, followed by kidneys, lungs and heart. Furthermore, the results provided evidence that EAI045 could be absorbed quickly and distributed widely in different tissue types.
Aims: Adipose-derived stem cells (ADSCs), a source of mesenchymal stem cells, are able to differentiate into numerous cell lineages, including epithelial and smooth muscle cells. The use of ADSCs in tissue engineering technology has become the most promising therapeutic approach for urethral reconstruction. This study aimed to explore the effect of IncRNA highly upregulated in liver cancer (HULC) on the induction of ADSCs to differentiate into epithelial and smooth-muscle-like cells. Methods: ADSCs were isolated from a male dog, and the expression of HULC in ADSCs was overexpressed by transfection with HULC expressing vector lentivirus. The transfected ADSCs were then incubated with 5 mu M ATRA or 2.5 ng/ml TGF-beta 1 and 5 ng/ml PDGF-BB for 21 days. The expression of epithelial differentiation and smooth-muscle-like differentiation markers were monitored. Besides, cross-regulation between HULC and BMP9 was detected in the differentiated epithelial cells and smooth-muscle-like cells. Results: HULC increased cell viability of ADSCs, but has no impact on ADSCs apoptosis. HULC promotes ADSCs to differentiate into epithelial and smooth-muscle-like cells, as evidenced by the increases in the expression of Uroplakin-II, AE1/AE3, alpha-SMA, SM-MHC, Calponin, and SM-22 alpha. In addition, HULC could positively regulate BMP9, and BMP9 silence abolished HULC-promoted ADSC's differentiation. Furthermore, HULC activated Wnt/beta-catenin pathway while deactivated Notch pathway. Conclusion: HULC was demonstrated to be a promoter during the epithelial and smooth-muscle-like differentiation of ADSCs via the BMP9/Wnt/beta-catenin/Notch network. This study provides the first in vitro evidence that HULC-based therapy could be a valuable approach to promote urethral reconstruction.
Aims: Long non-coding RNAs (lncRNAs) play key roles in cancers, yet their potential molecular mechanisms are not well understood. The objective of this study is to examine the expression, biological functions and mechanism of lncRNA CCAL in gastric cancer (GC). Methods: MTT and Colony formation assay were used to detect cell proliferation and the colony formation ability of gastric cancer cells. Wound healing, Migration, and invasion assay were respectively used to explore the migration, and invasion in gastric cancer cell lines. Realtime polymerase chain reaction (RT-PCR) was performed to determine the expression level of CCAL. Western Blot was used to determine the expression of related proteins. Results: In the present study, we found that CCAL was upregulated in gastric cancer cell lines. Patients whose tumors had high CCAL expression had a shorter overall survival than patients whose tumors had low CCAL expression. Overexpression CCAL promoted the proliferation, migration and invasion of GC by regulating the expression of myc. Conclusion: The present study reveals that CCAL is an oncogenic lncRNA that promotes the tumorigenesis and progression of GC.
Aim: MicroRNAs (miRs) are endogenous substances that act as important diagnostic and treatment targets in renal diseases. miR-146 plays an important role in the development of endotoxin tolerance through NF-kappa B pathway, but the underlying mechanism is not clearly understood. The aim of this study was to determine the molecular regulation and function of miR-146 and also the expression of miR-146 in an experimental model of renal ischemia reperfusion injury (IRI). Methods: IRI was induced in mouse by bilateral IRI for 45 min followed by reperfusion. The male mice were randomized as: sham, I/R, I/R+miR-146, and I/R+antago-miR-146 groups. Renal function, histological damage, and cell apoptosis were evaluated at 24 h after reperfusion. Results: Overexpression of miR-146 protected renal function. Renal cells with upregulated miR-146 had lower plasma levels of blood urea nitrogen (BUN) and creatinine, decreased apoptosis and active caspase-3 protein expressions. miR-146 was shown to have a role in renal IR injury. miR-146 has a protective effect on renal function and plays a significant role in apoptosis. IGSF1 acts as a target of miR-146. IGSF1 rescued the effects of miR-146 on renal IRI. miR-146 protected renal function by activation of PI3K/AKT. Conclusion: These findings suggest that miR-146 might regulate apoptosis and can cause injury in I/R via targeting IGSF1 and also exert renal protection property.
The dried seeds of Iris lactea Pall.var. chinensis (Fisch.) Koidz, an important traditional Chinese medicine, are regarded to have effects of clearing heat, eliminating dampness and pharyngitis and so on. It has been used in the treatment of jaundice, diarrhea, leucorrhea and carbuncles. Previous phytochemical studies of Iris species showed the presence of flavones, isoflavones, triterpenes and stilbenes. In our previous research, we isolated five known oligostilbenes, vitisin A, vitisin B, vitisin C, vitisin D, and cis-vitisin A were successfully isolated from Iris lactea for the first time. The aim of this study was to assess the effects of these oligostilbenes on the differentiation and adipogenes in 3T3-L1 cells. Our results showed that vitisin A, vitisin B, cis-vitisin A significantly inhibited adipocytes differentiation and reduced lipid accumulation in 3T3-L1 cells. In addition, vitisin A, vitisin B, cis-vitisin A strongly suppressed the expression levels of adipocyte-specific genes including peroxisome proliferator activated receptor-gamma (PPAR gamma), CCAAT/enhancer binding protein-alpha (C/EBP alpha) and adipocyte fatty acid binding protein 4 (FABP4). In contrast, vitisin C and vitisin D significantly promoted adipogenesis and increased intracellular lipid accumulation, while the two oligostilbenes markedly increased the expression of adipocyte marker genes. In the present study, we found that vitisin A, vitisin B and cis-vitisin A inhibit the adipogenesis and adipocytes differentiation by their influence on the expression of PPAR gamma, which leads to subsequenet downregulation of PPAR gamma mediated adipocyte-specific gene during adipogenesis.
High mobility group box 1 (HMGB1), a non-histone DNA-binding protein, is massively released into the extracellular space from neuronal cells after ischemic injury, initiates inflammatory response and aggravates brain tissue damage. Acetylpuerarin (AP), an acetylated derivative of puerarin, was reported to protect against cerebrovascular ischemia-reperfusion injury in rats through anti-inflammation. In the present study, we aim to investigate whether AP inhibited HMGB1 release in oxygen-glucose deprivation (OGD)-treated BV2 microglia. BV2 microglia viability after OGD with or without AP was measured by CCK-8 assay, apoptosis of BV2 microglia was determined by Hoechst 33258 staining and FITC-Annexin V/7-AAD staining. HMGB1 protein level and release was detected by western blotting and immunofluorescent FITC-staining. The results demonstrated that AP significantly rescued OGD-induced cell death and apoptosis in a dose-dependent manner. AP inhibited OGD-induced HMGB1secretion at the level of nuclear to cytoplasmic translocation, decreased cytoplasmic HMGB1 at protein level, and the effects showed dose-dependent. The findings suggest that AP can protect against OGD-induced cellular injury in BV2 microglia by inhibition of HMGB1 release.
Ganoderma lucidum extracts have shown antiepileptic effects in in vivo and in vitro studies. In this work, primary hippocampal neurons cultured in magnesium-free medium were used to study the neuroprotective effects of ganoderic acid A and B (GA-A and GA-B) on superoxide dismutase (SOD) activity and mitochondrial membrane potential, to improve our understanding of their antiepileptic effect. The activity of SOD was determined by the xanthine oxidase assay, the variations of mitochondrial membrane potential and cell apoptosis were measured by JC-1 fluorescent staining and flow cytometry. It was found that the SOD activity and mitochondrial membrane potential (118.84 U/mg protein and 244.08 Delta psi(m)) of the epileptic hippocampal neurons were significantly lower than control values (135.95 U/mg protein and 409.81 Delta psi(m)), associated with an increase of cell apoptosis (31.88% vs. 8.84%). These circumstances can be improved by treatment of GA-A/GA-B (for SOD, 127.15 +/- 3.82 / 120.52 +/- 4.30 U/mg protein; for membrane potential (Delta psi(m)), 372.35 / 347.28; and for cell apoptosis (%), 14.93 / 20.52). Results indicated that GA-A significantly improved SOD activity, while both GA-A/GA-B tranquillized the mitochondrial membrane potential of hippocampal neurons, and thereby protected these neurons by inhibiting apoptosis.
Cisplatin is an effective chemotherapeutic agent for osteosarcoma (OS) and has been shown to induce endoplasmic reticulum (ER) stress-associated apoptosis in human cancer cells. Ganglioside GD2-specific antibodies can inhibit tumor cell viability without involvement of the immune system. A recent study has shown that antiGD2 monoclonal antibody (mAb) 14G2a effectively inhibits the viability and invasiveness of human OS cells. In this study, we explored the effect of anti-GD2 mAb and cisplatin alone and in combination on ER stress-associated apoptosis in osteosarcoma cells. MG-63 and Saos-2 human OS cells were treated with cisplatin and/or an-GD2 mAb 14G2a for 48 hours. Cisplatin and 14G2a dose-dependently induced apoptosis in MG-63 and Saos-2 cells. They in combination induced 70%-77% of apoptosis in MG-63 cells and 79%-85% of apoptosis in Saos-2 cells, exhibiting a synergistic effect stronger than addition of their individual effects over the control level. Showing no significant effect on the expression of protein kinase RNA -like ER kinase (PERK), cisplatin and 14G2a exhibited a marked synergistic effect on inducing phosphorylation/activation of PERK, phosphorylation/inactivation of eukaryotic translation initiation factor 2 alpha (eIF2 alpha), expression of CHOP, in parallel to inducing the caspase-3 activity and apoptosis in MG-63 and Saos-2 cells. The effects were abolished by lentivirus-mediated knockdown of PERK. Particularly, PERK knockdown abolished 63% and 65% of the combined apoptotic effect of cisplatin and 14G2a on MG-63 and Saos-2 cells, respectively. In conclusion, this study provides the first evidence supporting that cisplatin and 14G2a synergize to induce ER stress-associated apoptosis in human OS cells through activating the PERK ER stress pathway by synergistically inducing phosphorylation/activation of PERK. Our findings add new insights into the pharmacologic effects of anti-GD2 mAb in anticancer treatment and suggest that cisplatin plus anti-GD2 mAb could be a new effective therapeutic strategy for OS.
The objective of this study was to prepare a new compound fenbendazole tablet containing 29.7 % fenbendazole, 1.50 % praziquantel and 0.059 % ivermectin for oral administration. The tablets were successfully prepared using mannitol as filler agent, polyvinyl polypyrrolidone as disintegrant, 5 % povidone (PVA(K30)) as a binder agent and magnesium stearate as lubricant. The appearance, hardness, fragility, time limit of disintegration and fenbendazole dissolution at 45 min all met the technical standards of the Ministry of Agriculture for the People's Republic of China. We used high performance liquid chromatography and electrospray-mass spectrometry for drug detection. Oral administration of 100 mg/kg fenbendazole, 5 mg/kg praziquantel and 0.2 mg/kg ivermectin using a non-compartmental model defined peak plasma concentrations (C-max) of 495, 826, 73 ng/mL, and 218 ng/mL for the metabolite oxfendazole, respectively. The area under the curve (AUC(last)) values for these drugs were 4653, 1045, 1971 and 5525 hxng/mL, respectively. This study enriches the pharmacokinetic data of compound fenbendazole tablets using dogs as a model system. The new tablet formulation was assimilated quickly and systemically and this study will be beneficial for the clinical application of parasite treatments in dogs.
Tamarix ramosissima is a traditional Chinese herbal medicine used for rheumatoid arthritis (RA) treatment in Northwest China. Chemical investigation of EtOH/H2O extracts of T. ramosissima led to the discovery of a new flavonol, ramosissimin (1), together with the known flavonoids compounds quercetin (2), quercetin-3'4'-dimethylether (3) and kaempferol (4). By means of high resolution electrospray ionization mass spectroscopy (HRESIMS) and 1D and 2D-NMR experiments, and after comparison with literature data, the structures of the compounds were determined. The effect of compound 1 on the viability of RA fibroblast-like synoviocytes (RA-FLS) was evaluated by MIT assay. Apoptosis-inducing effect of compound 1 in RA-FLS was further investigated by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay and activated caspase-3/7 level assessment using luminescent assay. The results revealed that ramosissimin displayed remarkable proliferation inhibitory effect in RA-FLS. Furthermore, compound 1 could significantly induce cellular apoptosis of RA-FLS and increase activated caspase-3/7 levels. It is suggested that ramosissimin may inhibit the proliferation of RA-FLS by inducing apoptosis.