[Mechanism of Qilongtian Capsules in treatment of acute lung injury based on network pharmacology prediction and experimental validation].

This study aimed to explore the mechanism of Qilongtian Capsules in treating acute lung injury(ALI) based on network pharmacology prediction and in vitro experimental validation. Firstly, UPLC-Q-TOF-MS/MS was used to analyze the main chemical components of Qilongtian Capsules, and related databases were used to obtain its action targets and ALI disease targets. STRING database was used to build a protein-protein interaction(PPI) network. Metascape database was used to conduct enrichment analysis of Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG). AutoDock software was used to perform molecular docking verification on the main active components and key targets. Then, the RAW264.7 cells were stimulated with lipopolysaccharide(LPS) for in vitro experiments. Cell viability was measured by MTT and ROS level was measured by DCFH-DA. NO content was measured by Griess assay, and IL-1ß, IL-6, and TNF-α mRNA expression was detected by RT-PCR. The predicted targets were preliminarily verified by investigating the effect of Qilongtian Capsules on downstream cytokines. Eighty-four compounds were identified by UPLC-Q-TOF-MS/MS. Through database retrieval, 44 active components with 589 target genes were screened out. There were 560 ALI disease targets, and 65 intersection targets. PPI network topology analysis revealed 10 core targets related to ALI, including STAT3, JUN, VEGFA, CASP3, and MMP9. KEGG enrichment analysis showed that Qilongtian Capsules mainly exerted an anti-ALI effect by regulating cancer pathway, AGE-RAGE, MAPK, and JAK-STAT signaling pathways. The results of molecular docking showed that the main active components in Qilongtian Capsules, including crenulatin, ginsenoside F_1, ginsenoside Rb_1, ginsenoside Rd, ginsenoside Rg_1, ginsenoside Rg_3, notoginsenoside Fe, notoginsenoside G, notoginsenoside R_1, notoginsenoside R_2, and notoginsenoside R_3, had good binding affinities with the corresponding protein targets STAT3, JUN, VEGFA, CASP3, and MMP9. Cellular experiments showed that Qilongtian Capsules at 0.1, 0.25, and 0.5 mg·mL~(-1) reduced the release of NO, while Qilongtian Capsules at 0.25 and 0.5 mg·mL~(-1) reduced ROS production, down-regulated mRNA expression of IL-1ß, IL-6, TNF-α, and inhibited the inflammatory cascade. In summary, Qilongtian Capsules may exert therapeutic effects on ALI through multiple components and targets.

Astragaloside IV attenuates high glucose-induced NF-κB-mediated inflammation through activation of PI3K/AKT-ERK-dependent Nrf2/ARE signaling pathway in glomerular mesangial cells.

Inflammation is a key contributor to diabetic kidney disease pathogenesis, including reactive oxidation stress (ROS)-mediated nuclear factor-κB (NF-κB) signaling pathway. In this study, we examined the effect of Astragaloside IV (AS-IV) on anti-inflammatory and anti-oxidative properties under high glucose (HG) condition and the potential mechanism in glomerular mesangial cells (GMCs). We showed that AS-IV concentration-dependently reduced GMCs proliferation, restrained ROS release and hydrogen peroxide content, and suppressed pro-inflammatory cytokines as well as pro-fibrotic factors expression, which were associated with the inhibition of NF-κB and nuclear factor-erythroid 2-related factor 2 (Nrf2) signaling activation. Accordingly, both NF-κB overexpression by using RNA plasmid and Nrf2 gene silencing by using RNA interference weakened the ability of AS-IV to ameliorate HG-induced oxidative stress, inflammation, and cell proliferation. Furthermore, phosphatidylinositide 3-kinases (PI3K)/serine/threonine protein kinase (Akt) and extracellular regulated protein kinases (ERK) signaling pathway regulated the process of AS-IV-induced Nrf2 activation and antioxidant capacity, which evidenced by using PI3K inhibitor LY294002 or ERK inhibitor PD98059 that largely abolished the AS-IV efficacy. Taken together, these results indicated that AS-IV protected against HG-induced GMCs damage by inhibiting ROS/NF-kB-induced increases of inflammatory cytokines, fibrosis biomarkers, and cell proliferation via up-regulation of Nrf2-dependent antioxidant enzyme expression, which were mediated by PI3K/Akt and ERK signaling pathway activation.

Mitochondrial FAD shortage in SLC25A32 deficiency affects folate-mediated one-carbon metabolism.

The SLC25A32 dysfunction is associated with neural tube defects (NTDs) and exercise intolerance, but very little is known about disease-specific mechanisms due to a paucity of animal models. Here, we generated homozygous (Slc25a32Y174C/Y174C and Slc25a32K235R/K235R) and compound heterozygous (Slc25a32Y174C/K235R) knock-in mice by mimicking the missense mutations identified from our patient. A homozygous knock-out (Slc25a32-/-) mouse was also generated. The Slc25a32K235R/K235R and Slc25a32Y174C/K235R mice presented with mild motor impairment and recapitulated the biochemical disturbances of the patient. While Slc25a32-/- mice die in utero with NTDs. None of the Slc25a32 mutations hindered the mitochondrial uptake of folate. Instead, the mitochondrial uptake of flavin adenine dinucleotide (FAD) was specifically blocked by Slc25a32Y174C/K235R, Slc25a32K235R/K235R, and Slc25a32-/- mutations. A positive correlation between SLC25A32 dysfunction and flavoenzyme deficiency was observed. Besides the flavoenzymes involved in fatty acid ß-oxidation and amino acid metabolism being impaired, Slc25a32-/- embryos also had a subunit of glycine cleavage system-dihydrolipoamide dehydrogenase damaged, resulting in glycine accumulation and glycine derived-formate reduction, which further disturbed folate-mediated one-carbon metabolism, leading to 5-methyltetrahydrofolate shortage and other folate intermediates accumulation. Maternal formate supplementation increased the 5-methyltetrahydrofolate levels and ameliorated the NTDs in Slc25a32-/- embryos. The Slc25a32K235R/K235R and Slc25a32Y174C/K235R mice had no glycine accumulation, but had another formate donor-dimethylglycine accumulated and formate deficiency. Meanwhile, they suffered from the absence of all folate intermediates in mitochondria. Formate supplementation increased the folate amounts, but this effect was not restricted to the Slc25a32 mutant mice only. In summary, we established novel animal models, which enabled us to understand the function of SLC25A32 better and to elucidate the role of SLC25A32 dysfunction in human disease development and progression.

Relevant effects of Taohong Siwu decoction on isolated rat aortic ring dependent on endothelium nitric oxide-cGMP pathway.

Using rat thoracic aortic rings to test the relaxing effects of the 95% ethanol extract and aqueous extract of Taohong Siwu decoction (THSW) on endothelium intact or endothelium removed aortic rings. Results showed that the 95% ethanol extract (0.1, 1, 10, 100, 1000 mg•L-1) and aqueous extract (0.1, 1, 10, 100, 1000 mg•L-1) of THSW were able to relax the intact endothelium aortic rings pre-contracted by 10-6 mol•L-1 PE. 10-4 mol•L-1 L-NAME and 10-5 mol•L-1 methylene blue both were able to inhibit the relaxation other than indomethacin. For the endothelium removed aortic rings, potassium channel blocker 3×10-3mol•L-1 tetraethylammonium chloride and 10-5 mol•L-1 glibenclamide had no effect on the relaxation effects caused by the 95% ethanol extract and aqueous extract of THSW. It could be concluded that the 95% ethanol extract and aqueous extract of THSW relax blood vessel by endothelium-dependent way.

Phenanthrenes from Juncus effusus.

A chemical investigation of the EtOAc-soluble fraction from the ethanol extract of the medullae of Juncus effusus led to the isolation of three new 9,10-dihydrophenanthrenes, juncuenins E-G (1-3); two new phenanthrenes, dehydrojuncuenins D-E (4-5); one new feruloylated glycoside (6); and one known 9,10-dihydrophenanthrene (7). The structures of these compounds were determined by analyzing their spectroscopic data. Metabolites 1-4 and 7 were further evaluated for their in vitro cytotoxic activities against seven human cancer lines (A549, MCF-7, BEL-7402, HeLa, COLO205, BGC-823, and SK-OV-3). Among them, compound 1 exhibited weak cytotoxicity against MCF-7 and HeLa cell lines. Compound 7 showed moderate cytotoxicity against MCF-7 and HeLa cell lines, with IC50 values of 9.17 and 19.6 µM, respectively.

A new acorane sesquiterpene from Lysionotus pauciflorus.

AIM: To study the chemical constituents of the whole plant of Lysionotus pauciflorus METHODS: Chromatographic separations on silica gel, Toyopearl HW-40F gel, and MCI gel were used to isolate the compounds. The structures were elucidated on the basis of spectral data. RESULTS: Three compounds were obtained and their structures were identified as 3, 10-dihydroxyacoronene (1), bis(2-butylhexyl)phthalate (2) and 3, 5-dimethoxy-4-hydroxy-trans-stilbene (3). CONCLUSION: Compound 1 is a new acorane sesquiterpene, this is the first report of acorane sesquiterpenes from the Lysionotus genus.

New dimeric phenanthrene and flavone from Spiranthes sinensis.

From the whole plant of Spiranthes sinensis, one novel dimeric phenanthrene, 2,2'-dihydroxy-5,5',7,7'-tetramethoxy-9,9',10,10'-tetrahydro-3,3'-biphenanthrene (1) and flavone 5-hydroxy-3,7-dimethoxy-4'-(1-hydroxy-3-methylbut-3-en-2-yloxy)-flavone (2) were isolated along with three other known flavonoids, which are reported for the first time from this species. Their structures were established after an extensive analysis by 1D and 2D NMR spectroscopy (NOESY, HSQC, and HMBC) as well as HR-TOF-MS.

Tomato TFT1 is required for PAMP-triggered immunity and mutations that prevent T3S effector XopN from binding to TFT1 attenuate Xanthomonas virulence.

XopN is a type III effector protein from Xanthomonas campestris pathovar vesicatoria that suppresses PAMP-triggered immunity (PTI) in tomato. Previous work reported that XopN interacts with the tomato 14-3-3 isoform TFT1; however, TFT1's role in PTI and/or XopN virulence was not determined. Here we show that TFT1 functions in PTI and is a XopN virulence target. Virus-induced gene silencing of TFT1 mRNA in tomato leaves resulted in increased growth of Xcv ΔxopN and Xcv ΔhrpF demonstrating that TFT1 is required to inhibit Xcv multiplication. TFT1 expression was required for Xcv-induced accumulation of PTI5, GRAS4, WRKY28, and LRR22 mRNAs, four PTI marker genes in tomato. Deletion analysis revealed that the XopN C-terminal domain (amino acids 344-733) is sufficient to bind TFT1. Removal of amino acids 605-733 disrupts XopN binding to TFT1 in plant extracts and inhibits XopN-dependent virulence in tomato, demonstrating that these residues are necessary for the XopN/TFT1 interaction. Phos-tag gel analysis and mass spectrometry showed that XopN is phosphorylated in plant extracts at serine 688 in a putative 14-3-3 recognition motif. Mutation of S688 reduced XopN's phosphorylation state but was not sufficient to inhibit binding to TFT1 or reduce XopN virulence. Mutation of S688 and two leucines (L64,L65) in XopN, however, eliminated XopN binding to TFT1 in plant extracts and XopN virulence. L64 and L65 are required for XopN to bind TARK1, a tomato atypical receptor kinase required for PTI. This suggested that TFT1 binding to XopN's C-terminal domain might be stabilized via TARK1/XopN interaction. Pull-down and BiFC analyses show that XopN promotes TARK1/TFT1 complex formation in vitro and in planta by functioning as a molecular scaffold. This is the first report showing that a type III effector targets a host 14-3-3 involved in PTI to promote bacterial pathogenesis.

Effects of Tongguan Capsule on post-myocardial infarction ventricular remodeling and cardiac function in rats.

OBJECTIVE: To observe the effects of Tongguan Capsule (TGC) on post-myocardial infarction ventricular remodeling and heart function in rats. METHODS: A rat model of acute myocardial infarction (AMI) was established by coronary ligation. Experimental rats were randomized to 4 groups including three model groups (Group A: captopril 5 mg/kg * day, n=7; Group B: TGC 10 g/kg * day, n=7; and Group C: placebo, n=8), and a sham-control group (Group D: blank control, n=6). Animals were treated for 4 weeks. The cardiac function of rats was assessed at the end of the experiment based on left ventricular ejection fraction (LVEF) and left ventricular short axis fractional shortening (LVFS) detected by colored echocardiography; meanwhile, the condition of ventricular remodeling was observed through the levels of left ventricular mass (LVM), plasma aldosterone (ALD), myocardial angiotensin II (Ang II) and myocardial collagen measurements. RESULTS: At the end of the experiment, LVEF and LVFS in Group A and B were improved significantly, while those in Group C were unchanged, the LVEF in Group A, B, C, and D was 0.57+/-0.46, 0.61+/-0.08, 0.36+/-0.55 and 0.76+/-0.02, respectively; and their LVFS was 0.31+/-0.52, 0.34+/-0.04, 0.23+/-0.57 and 0.45+/-0.03, respectively. The difference was statistically significant when comparing the two indexes in Group A and B with those in Group C and D (P<0.05). LVM, levels of plasma ALD and myocardial Ang II were lower in Group A and B than in Group C, but a comparison between Group A and B showed an insignificant difference in lowering LVM and ALD, while the lowering of Ang II was more significant in Group B than in Group A (754.7 +/- 18.7 pg/mL vs 952.6+/-17.6 pg/mL, P<0.05). Morphological examination showed that in Group A and B the swollen myocardial cells had shrunk, with regularly arranged myocardial fibers and decreased collagen proliferation, but the improvements in Group B were more significant. CONCLUSION: TGC could markedly improve the post-infarction ventricular remodeling and cardiac function in rats, showing that the efficacy was better than or equal to that of captopril.

Clinical effects of acupuncture combined with nimodipine for treatment of vascular dementia in 30 cases.

OBJECTIVE: To study the therapeutic effects of acupuncture combined with nimodipine for vascular dementia. METHODS: Acupuncture was applied at Baihui (GV 20), Shenshu (BL 23), Geshu (BL 17), and the points selected according to the midnight-noon, ebb-flow eight methods of the intelligent turtle, combined with the drug nimodipine. The treatment was continued for 8 consecutive weeks. RESULTS: Of the 30 cases treated, 6 cases were cured, 21 cases improved, and 3 cases failed, with a total effective rate of 90%. CONCLUSION: Acupuncture at Baihui (GV 20), Shenshu (BL 23), Geshu (BL 17), and the points selected according to the midnight-noon, ebb-flow eight methods of the intelligent turtle combined with the drug nimodipine can yield definite therapeutic effects for vascular dementia.

[Effects of top pruning on fruiting characters of Platycodon grandiflorum].

OBJECTIVE: To investigate the effects of top pruning on fruiting characters of Platycodon grandiflorum, and find the suitable stage, in which seed growth and development furtherly. METHOD: One-year old seedlings were chosen and planted in field. Plant height, branching number, fruit number per plant, 1000 grains weight were measured during growth and development period, respectively. RESULT: The treatment of top pruning postponed in turn the flowering date, lowered the plant heights and the fruit number per plant, increased the branching number and influenced significantly on 1000 grains weights. CONCLUSION: The suitable stage of top pruning for producing seeds was from June 20th to July 5th.

[Study on changes of TCM syndrome in patients with coronary heart disease before and after intervention treatment].

OBJECTIVE: To observe the changing laws of TCM syndrome type in patients with coronary heart disease (CHD) before and after intervention treatment (IT) and to explore the influence of IT on TCM syndrome type. METHODS: The TCM syndrome type of 71 patients with "Chest-Bi" was differentiated before and after percutaneous coronary intervention (PCI) treatment, of which the most common syndrome types were qi deficiency, yang deficiency, yin deficiency, qi stagnation, blood stasis, phlegm, cold coagulation, heat-syndrome, etc. RESULTS: Before PCI treatment, syndrome types of blood stasis (53 cases, 74.6%), qi deficiency (46 cases, 64.8%), and phlegm (28 cases, 39.4%) were the commonest, while there were 12 cases of qi stagnation (16.9%) and 12 cases of cold coagulation (16.9%); One week after PCI treatment, the most commonly seen types were blood stasis (47 cases, 66.2%), qi deficiency (39 cases, 54.9%) and phlegm (23 cases, 32.4%), while qi stagnation (2 cases, 2.8%) and cold coagulation (1 case, 1.4%) were also found; One month after PCI, qi deficiency (47 cases,85.4%), blood stasis (40 cases,72.7%), phlegm (31 cases, 56.4%) were the most commonly seen types. Comparison of the syndrome types between before and after PCI showed that the syndromes of qi deficiency and phlegm were progressively aggravating, while syndromes of qi stagnation and cold coagulation were alleviated after PCI. CONCLUSION: Although PCI treatment could relieve patients' symptoms of excess in superficiality, it can't radically change the pathogenetic nature of CHD, namely, the deficiency in origin and excess in superficiality, which indicates that one should pay full attention to the importance and necessity of CHD after PCI treatment.

[Analysis of RAPD on medicinal plants of Codonopsis pilosula].

OBJECTIVE: To study genetic diversity and genetic relationship among 9 cultivated and 4 nature populations of Codonopsis pilosula and C. pilosula var. modesta form Gansu Province at DNA level. METHODS: DNA of 217 individuals was amplified by 14 decamer primers from 200 radom primers. The polymorphic bands of RAPD were counted. And the results were analyzed by clustering analysis NTSYS-PC software. RESULTS: A total of 125 loci were detected by 14 arbitrary primers, among which 109 and 106 loci were polymorphic in C. pilosula and C. pilosula var. modesta respectively, and the percentage of polymorphic bands (PPB) were 87.20% and 84. 80%. The cluster analysis showed that 13 populations were divided into 2 groups. The one included 8 cultivated populations of C. pilosula, the other group consisted of a cultivated population and 4 nature populations of C. pilosula var. modesta. CONCLUSION: The RAPD results indicated that there was abundant genetic diversity in cultivated populations of C. pilosula and C. pilosula var. modesta. The genetic differentiation among 8 cultivated populations of C. pilosula was very small and related to their geographical distance. Only the cultivation in Wen County originated from C. pilosula var. modesta, while the other populations cultivated in Gansu Province belongs to C. pilosula.