Mechanism of Magnolia Volatile Oil in the Treatment of Acute Pancreatitis Based on GC-MS, Network Pharmacology, and Molecular Docking.

Objective: Magnoliae officinalis cortex (MOC) is one of the most frequently used traditional Chinese medicine (TCM) for the treatment of acute pancreatitis (AP). Magnolia volatile oil (MVO) is considered to be one of the main active ingredients in MOC for AP treatment. However, the underlying mechanism of MVO in AP therapy is unknown. Methods: An integrated strategy of gas chromatography-mass spectrum (GC-MS), network pharmacology, and molecular docking simulation was employed to predict underlying mechanism of MVO in AP treatment. First, the compounds of MVO were identified by GC-MS, and the targets of the identified characteristic compounds were collected from several databases, as well as AP-related targets. Next, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis were carried out to obtain the mechanism. Moreover, the binding activity between core therapeutic targets and their corresponding compounds was evaluated by molecular docking simulation. Results: GC-MS results showed a total of 35 compounds that appeared in at least 18 out of 20 chromatograms were considered as characteristic compounds of MVO, and 33 compounds of those were identified. Network analysis demonstrated that 33 compounds regulated 142 AP-related targets. Of those, 8 compounds (α-eudesmol, γ-eudesmol, (-)-terpinen-4-ol, terpineol, hinesol, linalool, borneol, and ß-eudesmol) and 8 targets (TNF, IL-1ß, PPARγ, PPARα, PTGS2, NCOA1, CNR1, and ESR1) have a close relationship with AP treatment and were recognized as the key active compounds and the core therapeutic targets, respectively. The 142 targets were involved in both inflammation and calcium overload-related biological pathways, such as neuroactive ligand-receptor interaction, estrogen, MAPK, and calcium signaling pathway. Moreover, molecular docking simulation indicated that the 8 core therapeutic targets strongly interacted with their corresponding compounds. Conclusions: In summary, the present study elucidated that the efficacy of MVO in AP treatment might be attributed to anti-inflammation and inhibition of calcium overload through multicomponents and multitargets.

Visual detection of aflatoxin B1 and zearalenone via activating a new catalytic reaction of "naked" DNAzyme.

Here, we have found for the first time that the catalytic activity of "naked" DNAzyme, a single-stranded G-quadruplex DNAzyme (S.DNAzyme), can be modulated by aflatoxin B1 (AFB1) and zearalenone (ZEN). In fact, S.DNAzyme can mimic the activity of horseradish peroxidase to perform oxidation of 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) to colored ABTS˙+ (dark green). This catalytic activity can be inhibited upon addition of AFB1, leading to color fade of the solution. But with ZEN, the solution color will change to a yellowish-brown or yellow color. Thus, we have exploited this finding to achieve label-free, naked-eye detection of AFB1 or ZEN, and have explored the possible detection mechanisms. This approach is able to detect AFB1 and ZEN concentrations as low as 0.18 µM and 0.29 µM, respectively. Even in real samples, the limit of detection values of these two analytes are lower than 3 µM. Notably, S.DNAzyme cross-reacted with three other tested aflatoxins, including aflatoxin B2, aflatoxin G1 and aflatoxin M1, revealing the potential for a broadly applicable method to identify the family of aflatoxins. The AFB1- or ZEN-triggered new catalytic reaction between "naked" DNAzyme and ABTS may offer new opportunities for on-site visual detection of mycotoxins.

Comparison of Volatile Oil between the Ligusticum sinese Oliv. and Ligusticum jeholense Nakai et Kitag. Based on GC-MS and Chemical Pattern Recognition Analysis.

Ligustici Rhizoma et Radix (LReR) is the dried rhizomes and roots of Ligusticum sinese Oliv. (LS) or Ligusticum jeholense Nakai et Kitag. (LJ). However, in the market, LS and LJ are frequently confused with each other. Since the volatile oils are both the main active components and quality control indicators of LReR, a strategy combining gas chromatography-mass spectrometry (GC-MS) and chemical pattern recognition (CPR) was used to compare the volatile components of LJ and LS. Total ion chromatography (TIC) revealed that phthalides (i.e., neocnidilide) and phenylpropanoids (i.e., myristicin) could be thought of as the most critical components in the volatile oils of LJ and LS, respectively. In addition, the chemical components of the volatile oils in LJ and LS were successfully distinguished by hierarchical cluster analysis (HCA) and principal component analysis (PCA). Moreover, two quality markers, including myristicin and neocnidilide, with a very high discriminative value for the classification of LJ and LS, were found by orthogonal partial least squares discriminant analysis (OPLS-DA). The relative contents of myristicin and neocnidilide were 10.86 ± 6.18% and 26.43 ± 19.63% for LJ, and 47.43 ± 12.66% and 2.87 ± 2.31% for LS. In conclusion, this research has developed an effective approach to discriminating LJ and LS based on volatile oils by combining GC-MS with chemical pattern recognition analysis.

Comparison of Volatile Oil between the Fruits of Amomum villosum Lour. and Amomum villosum Lour. var. xanthioides T. L. Wu et Senjen Based on GC-MS and Chemometric Techniques.

Fructus Amomi (FA) is usually regarded as the dried ripe fruit of Amomum villosum Lour. (FAL) or Amomum villosum Lour. var. xanthioides T. L. Wu et Senjen (FALX.). However, FAL, which always has a much higher price because of its better quality, is often confused with FALX. in the market. As volatile oil is the main constituent of FA, a strategy combining gas chromatography-mass spectrometry (GC-MS) and chemometric approaches was applied to compare the chemical composition of FAL and FALX. The results showed that the oil yield of FAL was significantly higher than that of FALX. Total ion chromatography (TIC) showed that cis-nerolidol existed only in FALX. Bornyl acetate and camphor can be considered the most important volatile components in FAL and FALX., respectively. Moreover, hierarchical cluster analysis (HCA) and principal component analysis (PCA) successfully distinguished the chemical constituents of the volatile oils in FAL and FALX. Additionally, bornyl acetate, α-cadinol, linalool, ß-myrcene, camphor, d-limonene, terpinolene and borneol were selected as the potential markers for discriminating FAL and FALX. by partial least squares discrimination analysis (PLS-DA). In conclusion, this present study has developed a scientific approach to separate FAL and FALX. based on volatile oils, by GC-MS combined with chemometric techniques.

Treatment of symptomatic middle cerebral artery stenosis with balloon-mounted stents: long-term follow-up at a single center.

OBJECTIVE: Primary stenting is a well-established treatment for coronary artery disease and has been applied to symptomatic intracranial stenosis in selected patients. This study reports a large case series of middle cerebral artery revascularization using balloon-mounted coronary stents. METHODS: The series consisted of 113 consecutive patients with symptomatic middle cerebral artery stenosis and more than 70% occlusion who underwent intracranial stenting at a single center from 2001 to 2006. Technical success, periprocedural complications, recurrent symptoms, and restenosis were retrospectively reviewed. Risk factors for restenosis were analyzed using logistic regression and the chi(2) test. RESULTS: The mean age of the patients was 48 +/- 11 years (range, 25-79 years). Seventy-three patients presented with transient ischemic attacks, and 40 patients were diagnosed with acute stroke. The technical success rate was 96.46%. The mean stenosis was reduced from 80.83 +/- 8.77% to 3.71 +/- 8.15%. The rate of stroke and death within 30 days was 4.42%. Eighty-nine patients were followed for an average period of 29 +/- 16 months (range, 9 months-5 years). Seventy-nine vessels were followed with transcranial Doppler and 36 vessels with angiography or computed tomographic angiography. Recurrent stroke or transient ischemic attacks occurred in 6 patients (6.74%). The restenosis rate was 20.25%. Restenosis was associated with diabetes and hyperlipidemia but not with age, sex, hypertension, or drug-eluting stent. CONCLUSION: Primary stenting of symptomatic middle cerebral artery stenosis can be performed with high success and low complication rates. Randomized clinical studies are warranted to compare the safety and efficacy of various endovascular revascularization techniques with maximal medical therapy in patients with symptomatic intracranial atherosclerosis.

[Studies on chemical constituents in n-butyl alcohol extract of roots of Actinidia deliciosa in Guangxi].

OBJECTIVE: To study the chemical constituents of n-butyl alcohol extract in the roots of Actinidia deliciosa in Guangxi. METHOD: The constituents were separated with various chromatographic techniques and their structures were elucidated by means of physicochemical properties and the analysis of their spectral data. RESULT: Six compounds were isolated and identified as eriantic acid B (1), 2alpha, 3beta, 24-trihydroxyursa-12-en-28-oic acid (2), 2alpha, 3alpha, 24-trihydroxyursa-12-en-28-oic acid (3), 2alpha, 3alpha, 23-tri-hydroxyursa-12, 20 (30)-dien-28-oic acid (4), 2alpha, 3alpha, 24-trihydroxyursa-12, 20 (30)-dien-28-oic acid (5), n-butyl-O-beta-D-fruto-pyranoside (6). CONCLUSION: Compounds 1-4, 6 were obtained from this plant for the first time. Compound 6 was obtained from the genus Actinidia for the first time.

[Screening of polypeptides interacting with Rel homology domain of NF-kappaB p50 subunit].

AIM: To obtain the polypeptides interacting with NF-kappaB p50 Rel homology domain (RHD) by yeast two-hybrid technique. METHODS: Using NF-kappaB p50 RHD as bait, the polypeptides interacting with NF-kappaB p50 RHD were screened from a 16-peptide cDNA library by yeast two-hybrid technique. The false positive clones were screened out but the positive clone were identified by beta-GAL assay, yeast mating, and one to one yeast two-hybrid method. RESULTS: 8 positive clones were obtained. The gene sequences of the eight polypeptides were different from each other and their homologous proteins were not found in GenBank. CONCLUSION: 8 novel polypeptides interacting with NF-kappaB p50 RHD were obtained, but their functions need to be validated in further experiments.

[Cloning and identification of NF-kappaB p50 Rel homology domain and detection of its autonomous reporter gene activity].

AIM: To clone NF-kappaB p50 Rel homology domain (RHD) gene and construct the "bait" vector in yeast two-hybrid system, and detect the yeast cell toxicity and autonomous reporter gene activity of target gene. METHODS: Total RNA was extracted from human peripheral blood mononuclear cells and NF-kappaB p50 RHD gene was amplified by RT-PCR, and cloned into pGBKT7. The recombinant plasmid was transformed into yeast AH109. The growth condition of the transformants was observed in the selected medium SD/-Trp. The reporter gene activity of target gene in the yeast cells was verified by filter blotting. RESULTS: Using restriction enzyme digestion analysis and PCR, the length of inserted gene was confirmed correct. Sequencing result indicated that the sequence of the inserted gene and its open reading frame were completely correct, and then the recombinant plasmid was named pGBKT7-p50. p50 RHD gene had neither autonomous reporter gene activity nor yeast cytotoxicity. CONCLUSION: As a bait plasmid, pGBKT7-p50 could be used in yeast two-hybrid system to screen and capture the polypeptides which interact with p50 RHD.

[Progress in locked nucleic acid research].

Locked nucleic acid (LNA) is a novel oligonucleotide analogue in which 2'-O and 4'-C positions in the b-D-ribofuranosyl ring are joined via an O-methylene, S-methylene or amino-methylene moiety, locked in a C3'-endol3E north (N)-type furanose conformation. LNAs posses many properties, such as extraordinarily high hybridize affinities for complementary DNA/RNA sequences, remarkable antisense activity, nuclease resistance, good aqueous solubility and none detactable toxicity, et al. LNAs is a most promising molecule for development of diagnostics and therapeutics. For example, apply LNAs to single--nucleotide polymorphisms genotyping, telomerase activity have been efficiently suppressed by LNA oligomers, efficient cleavage of highly structured RNA has been achieved using LNA-modified DNAzymes (LNAzymes), and so on.