Ethanol extract of Vanilla planifolia stems reduces PAK6 expression and induces cell death in glioblastoma cells.

Glioblastoma multiforme (GBM) is a malignant tumour with a poor prognosis. Therefore, potential treatment strategies and novel therapeutic targets have gained increased attention. Our data showed that the ethanol extract of Vanilla planifolia stem (VAS) significantly decreased the viability and the colony formation of GBM cells. Moreover, VAS induced the cleavage of MAP1LC3, a marker of autophagy. Further RNA-seq and bioinformatic analysis revealed 4248 differentially expressed genes (DEGs) between VAS-treated GBM cells and the control cells. Protein-protein interactions between DEGs with fold changes less than -3 and more than 5 were further analysed, and we found that 16 and 9 hub DEGs, respectively, were correlated with other DEGs. Further qPCR experiments confirmed that 14 hub DEGs was significantly downregulated and 9 hub DEGs was significantly upregulated. In addition, another significantly downregulated DEG, p21-activated kinase 6 (PAK6), was correlated with the overall survival of GBM patients. Further validation experiments confirmed that VAS significantly reduced the mRNA and protein expression of PAK6, which led to the abolition of cell viability and colony formation. These findings demonstrated that VAS reduced cell viability, suppressed colony formation and induced autophagy and revealed PAK6 and other DEGs as potential therapeutic targets for GBM treatment.

Meis1, Hi1α, and GATA1 are integrated into a hierarchical regulatory network to mediate primitive erythropoiesis.

During development, erythroid cells are generated by two waves of hematopoiesis. In zebrafish, primitive erythropoiesis takes place in the intermediate cell mass region, and definitive erythropoiesis arises from the aorta-gonad mesonephros. TALE-homeoproteins Meis1 and Pbx1 function upstream of GATA1 to specify the erythroid lineage. Embryos lacking Meis1 or Pbx1 have weak gata1 expression and fail to produce primitive erythrocytes. Nevertheless, the underlying mechanism of how Meis1 and Pbx1 mediate gata1 transcription in erythrocytes remains unclear. Here we show that Hif1α acts downstream of Meis1 to mediate gata1 expression in zebrafish embryos. Inhibition of Meis1 expression resulted in suppression of hif1a expression and abrogated primitive erythropoiesis, while injection with in vitro-synthesized hif1α mRNA rescued gata1 transcription in Meis1 morphants and recovered their erythropoiesis. Ablation of Hif1α expression either by morpholino knockdown or Crispr-Cas9 knockout suppressed gata1 transcription and abrogated primitive erythropoiesis. Results of chromatin immunoprecipitation assays showed that Hif1α associates with hypoxia-response elements located in the 3'-flanking region of gata1 during development, suggesting that Hif1α regulates gata1 expression in vivo. Together, our results indicate that Meis1, Hif1α, and GATA1 indeed comprise a hierarchical regulatory network in which Hif1α acts downstream of Meis1 to activate gata1 transcription through direct interactions with its cis-acting elements in primitive erythrocytes.

Evodiamine has therapeutic efficacy in ulcerative colitis by increasing Lactobacillus acidophilus levels and acetate production.

Emerging evidence implicates gut microbiota have an important role in ulcerative colitis (UC). Previous study indicated that Evodiamine (EVO) can alleviate colitis through downregulating inflammatory pathways. However, specific relationship between EVO-treated colitis relief and regulation of gut microbiota is still unclear. Here, our goal was to determine the potential role of gut microbiota in the relief of UC by EVO. By using pathology-related indicators, 16S rRNA sequencing and metabolomics profiling, we assessed the pharmacological effect of EVO on dextran sulfate sodium (DSS)-induced colitis rats as well as on the change of gut microbiota and metabolism. Fecal derived from EVO-treated rats was transplanted into colitis rats to verify the effect of EVO on gut microbiota, and 'driver bacteria' was found and validated by 16S rRNA sequencing, metagenome and qRT-PCR. The effect of Lactobacillus acidophilus (L. acidophilus) was investigated by vivo experiment, microbiota analysis, Short-chain fatty acids (SCFAs) quantification and colon transcriptomics. EVO reduced the susceptibility to DSS-induced destruction of epithelial integrity and severe inflammatory response, and regulated the gut microbiota and metabolites. Fecal Microbiota Transplantation (FMT) alleviated DSS-induced colitis, increased the abundance of L. acidophilus and the level of acetate. Furthermore, gavaged with L. acidophilus reduced pro-inflammatory cytokines, promoted the increase of goblet cells and the secretion of antimicrobial peptides, regulated the ratio of Firmicutes/Bacteroidetes and increased the level of acetate. Our results indicated that EVO mitigation of DSS-induced colitis is associated with increased in L. acidophilus and protective acetate production, which may be a promising strategy for treating UC.

Simultaneous determination of gallic acid, methyl gallate, and 1,3,6-tri-O-galloyl-ß-d-glucose from Turkish galls in rat plasma using liquid chromatography-tandem mass spectrometry and its application to pharmacokinetics study.

Turkish galls (TG) is a traditional Uygur medicine typically used in clinics for dental disease and chronic ulcerative colitis. In this study, a novel liquid chromatography-tandem mass spectrometry method was developed and validated for the simultaneous quantification of gallic acid, methyl gallate, and 1,3,6-tri-O-galloyl-ß-d-glucose in rat plasma, which are the major bioactive compounds of TG. After a feasible protein precipitation using acetonitrile for sample preparation, chromatographic separation was performed with a BDS Hypersil C18 column (2.1 × 100 mm, 5 µm) at 30°C, and water containing 10 mmol of ammonium acetate and acetonitrile was used as the mobile phase with a flow rate of 0.3 mL/min. The MS detector was operated in the selective reaction monitoring with negative-ionization mode. The results of the method validation, including selectivity, linearity, accuracy, precision, extraction recovery, matrix effect, and stability of the compounds in the biosamples, were all within the current acceptance criteria. The established method was successfully applied to the pharmacokinetics study of three analytes in rats after an oral administration of TG extract and laid the foundation for studying the active components and mechanism of TG in vivo.

[Research advances on astragaloside-IV in treatment of diabetes mellitus and its complications pharmacological effects].

Diabetes has become a global public health problem that seriously threatens human health. Traditional Chinese medicine, the characteristics of the role of multiple targets, has a unique advantage in the prevention and treatment of diabetes mellitus and its complications. Astragaloside-Ⅳ (AS-Ⅳ), one of the main activities of Astragalus membranaceus, has a series of pharmacological effects including improvement in the function of endothelial cells and neovascularization, anti-inflammatory, antioxidant, regulating energy metabolism, protectionnervous, anti-cancer and so on. In this paper, AS-Ⅳ to prevent and treat diabetes and its complications has been reviewed, which has effect on lowering blood sugar, lowering blood pressure, improving insulin resistance, inhibiting inflammatory reaction and oxidative stress. Additionally, it also can improve the diabetic animal and cell model of diabetic vascular disease, diabetic nephropathy, diabetic retinopathy, diabetic peripheral neuropathy, diabetic cardiomyopathy and other pathological damages. AS-Ⅳ may be a potential active substance for the treatment of diabetes and its complications.

Effect of Ginkgo biloba extract on lipopolysaccharide-induced anhedonic depressive-like behavior in male rats.

The peripheral administration of lipopolysaccharide (LPS) induces depressive-like behavior. Anhedonia is a core symptom of depression, defined as a loss of the capacity to experience pleasure. The present study used the sucrose preference test to investigate the influence of Ginkgo biloba extract (EGb 761) on LPS-induced anhedonia in male rats. The animals were randomly divided into four groups: (I) vehicle + saline, (II) vehicle + LPS, (III) EGb 761 + saline, and (IV) EGb 761 + LPS. Saline or LPS (100 µg/kg) was administered intraperitoneally 2 h before the sucrose preference test. Sucrose consumption was recorded 2, 4, 6, 13, and 24 h after 100 µg/kg of LPS or saline injection in the dark phase of the light/dark cycle. Dopamine and serotonin levels in the nucleus accumbens were measured. Our results indicated that the vehicle + LPS group exhibited a significant decrease in sucrose intake compared with the vehicle + saline group. The EGb 761 + LPS group showed more sucrose and food consumption than the vehicle + LPS group. Additionally, compared with the EGb 761 + LPS group, the vehicle + LPS group had less dopamine levels in the nucleus accumbens. Treatment with EGb 761 had no effect on water intake. Our results suggest that EGb 761 may be useful for reducing anhedonic depressive-like behavior.

Integrating network pharmacology and transcriptomics to study the potential mechanism of Jingzhi Niuhuang Jiedu tablet in rats with accumulation of heat in the lungs and stomach.

ETHNOPHARMACOLOGICAL RELEVANCE: Accumulation of heat in the lungs and stomach (AHLS) is an important syndrome within the realm of traditional Chinese medicine (TCM). It is the fundamental reason behind numerous illnesses, including mouth ulcers, dermatological conditions, acne, and pharyngitis. Jingzhi Niuhuang Jiedu tablet (JN) serves as the representative prescription for treatment of AHLS clinically. However, the effective components and mechanism of JN's impact on AHLS remain unexplored. AIM OF THE STUDY: The objective of this research was to analyze the effective components of JN and investigate the therapeutic effect and potential mechanism of JN on AHLS. MATERIALS AND METHODS: The effective compounds of JN extract were analyzed and identified using UHPLC-Q-Exactive/HRMS. Utilizing network pharmacology to investigate JN's multi-target, multi-pathway process in treating AHLS. Subsequently, anti-inflammatory activities of JN extract were evaluated in the RAW264.7 cells stimulated by lipopolysaccharide (LPS). In addition, a rat AHLS model induced by LPS and dried ginger was established. Pathological changes in rat lung and stomach tissues observed by HE staining and Masson's trichrome staining. Additionally, the expression of TNF-α, IL-6, and IL-1ß in bronchoalveolar lavage fluid (BALF) was identified through the ELISA assay. For a deeper understanding of how JN might affect AHLS, transcriptomics was utilized to examine differential genes and their underlying mechanisms. Concurrently, techniques like quantitative polymerase chain reaction (q-PCR), immunofluorescence, and western blotting (WB) were employed to confirm various mRNA and protein expression, including Il17ra, Il17re, IL-17A, IL-1ß, IL-6, PPARγ, PGC1-α and UCP1. RESULTS: We identified 178 potential effective components in the JN extract. Network pharmacology analysis showed that the 144 components in JN act on 200 key targets for the treatment of AHLS by suppressing inflammation, regulating energy metabolism, and gastric function. In addition, JN suppressed the LPS-stimulated generation of NO, TNF-α, IL-1ß, and IL-6 in RAW264.7 cells. And JN treatment effectively alleviated lung and stomach injury and reduced inflammation in rats. Analysis of RNA-seq from lung tissues revealed JN's substantial control over crucial genes in the IL-17 signaling pathway, including Il1b and Il17ra. Likewise, RNA sequencing of stomach tissues revealed that JN markedly decreased crucial genes in the Thermogenesis pathway, including Ppargc1a and Ppara. Additional experimental findings confirmed that treatment with JN significantly reduced the expression levels of mRNA (Il17ra, Il17re, Il1b, Ppargc1a and Ucp1), and the expression levels of protein (IL-17A, IL-1ß, IL-6, PPARγ, PGC1-α and UCP1). CONCLUSION: This study not only analyzes the effective components of JN but also reveals that JN could effectively ameliorate AHLS by inhibiting IL-17 signaling pathway and Thermogenesis pathway, which provides evidence for subsequent clinical studies and drug development.

[Analysis on Serology and FUT1 Gene of Para-Bombay Phenotype in Zhangzhou Area].

OBJECTIVE: To investigate blood group serological manifestations and molecular mechanism of para-Bombay phenotype. METHODS: The serological manifestations of para-Bombay phenotype was identified by serological assay. Molecular mechanism of para-Bombay phenotype was detected by PCR amplification. RESULTS: Eighteen cases of para-Bombay phenotype were detected through serology and PCR, including 5 cases of Amh, 4 cases of Bmh, and 10 cases of Omh. Eight cases produced anti-HI antibody. CONCLUSIONS: Blood group serology combined with molecular biology can accurately identify para-Bombay phenotype. Special transfusion strategy should be adopted for individual with para-Bombay phenotype to avoid hemolytic reactions.

Phosphine/thiolate-containing dinitrosyl cobalt complexes (DNCCs): synthesis, characterization, interconversion, X-ray diffraction identification and NO release.

Cobalt carbonyl/nitrosyl complexes, (PPh3)(CO)2Co(NO) (1) and (PPh3)2(CO)Co(NO) (2), were obtained by reacting (CO)3Co(NO) with one equiv. and two equiv. of PPh3, respectively. The process of isoelectronic replacement of CO with NO+ resulted in the formation of a cationic complex {Co(NO)2}10 [(PPh3)2Co(NO)2][BF4] (3). Complex (PPh3)(SPh)Co(NO)2 (4), which contains a thiophenolate ligand, was synthesized by ligand exchange of complex 3 with [PPh4][SPh] in a 1 : 1 molar ratio in THF solution. The addition of one equiv. of [PPh4][SPh] to complex 4 led to the formation of complex [PPh4][(SPh)2Co(NO)2] (5). The interconversions among complexes 1-5 were substantiated with the application of IR spectroscopy and X-ray single-crystal diffraction techniques. Notably, complex 4 exhibited commendable NOs (nitric oxide species: NO+/˙NO/NO-) transfer capabilities in the presence of [Fe(TPP)Cl] (5,10,15,20-tetraphenyl-21H,23H-porphine iron(III) chloride).

Effect of De-Twinning on Tensile Strength of Nano-Twinned Cu Films.

Tensile tests were carried on the electroplated Cu films with various densities of twin grain boundary. With TEM images and a selected area diffraction pattern, nano-twinned structure can be observed and defined in the electroplated Cu films. The density of the nano-twin grain structure can be manipulated with the concentration of gelatin in the Cu-sulfate electrolyte solution. We found that the strength of the Cu films is highly related to the twin-boundary density. The Cu film with a greater twin-boundary density has a larger fracture strength than the Cu film with a lesser twin-boundary density. After tensile tests, necking phenomenon (about 20 µm) occurred in the fractured Cu films. Moreover, by focused ion beam (FIB) cross-sectional analysis, the de-twinning can be observed in the region where necking begins. Thus, we believe that the de-twinning of the nano-twinned structure initiates the plastic deformation of the nano-twinned Cu films. Furthermore, with the analysis of the TEM images on the nano-twinned structure in the necking region of the fractured Cu films, the de-twinning mechanism attributes to two processes: (1) the ledge formation by the engagement of the dislocations with the twin boundaries and (2) the collapse of the ledges with the opposite twin-boundaries. In conclusion, the plastic deformation of nano-twinned Cu films is governed by the de-twinning of the nano-twinned structure. Moreover, the fracture strength of the nano-twinned Cu films is proportional to the twin-boundaries density.

[Analysis of Related Factors of Adverse Transfusion Reactions].

OBJECTIVE: To investigate the related factors of adverse reactions of blood transfusion, and clinical precautions so as to reduce the adverse reactions. METHODS: Data of 2108 patients with allogeneic transfusion in our hospital from January 2017 to June 2017 collected and analyzed. RESULTS: These patients received 15 244 time of blood transfusion, and 213 time of adverse reactions occurred in 178 patients in totality, the incidence is 1.4%, and there was no significant difference between the male (1.31%) and female (1.53%). The main type of transfusion reaction were allergy (73.23%), FNHTR (11.74%) and TACO (10.80%). Among all kinds of blood components, the incidence of adverse reactions of apheresis platelet transfusion was the highest (4.31%), significantly higher than that of cryoprecipitate and other blood components. The incidence rate of adverse reactions of blood transfusion in the hematopathy patients was 2.56%, significantly higher than that of immune diseases (1.48%), cancer diseases (1.28%) and other diseases (1.08%), (P＜0.01). The rate of transfusion history of apheresis platelets was 42.67% (the incidence of adverse reactions was 4.31%), significantly higher than other groups (P＜0.01); the rate of transfusion history of cryoprecipitate was 4.11% (the incidence of adverse reaction was 0.45%), significantly lower than other groups (P＜0.05). Among the disease types, the rate of transfusion history in the hematopathy patients was 48.79% (the incidence of adverse reaction was 2.56%), significantly higher than that of other groups (P＜0.01). The incidence of drug allergy in patients with the adverse reactions to blood transfusion was 11.25%, significantly higher than that of patients without adverse reactions (4.71%) (P＜0.01). CONCLUSION: The main risk factors of adverse reactions of blood transfusion are as follows: blood varieties, disease type, transfusion history and drug allergy history. For the patients with transfusion, multiple factors should be controlled, so as to reduce the adverse reactions.