Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry combined with chemometrics to identify the origin of Chinese medicinal materials.

Geographical origin and authenticity are two core factors to promote the development of traditional Chinese medicine (TCM) herbs perception in terms of quality and price. Therefore, they are important to both sellers and consumers. Herein, we propose an efficient, accurate method for discrimination of genuine and non-authentic producing areas of TCM by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Take Atractylodes macrocephala Koidz (AMK) of compositae as an example, the MALDI-TOF MS spectra data of 120 AMK samples aided by principal component analysis-linear discriminant analysis (PCA-LDA), partial least squares discriminant analysis (PLS-DA) and random forest (RF) successfully differentiated Zhejiang province, Anhui province and Hunan province AMK according to their geographical location of origin. The correct classification rates of test set were above 93.3%. Furthermore, 5 recollected AMK samples were used to verify the performance of the classification models. The outcome of this study can be a good resource in building a database for AMK. The combined utility of MALDI-TOF MS and chemometrics is expected to be expanded and applied to the origin traceability of other TCMs.

Application of Different Doses of Tranexamic Acid Plus Traditional Chinese Medicine in Hip Arthroplasty in Patients with Diabetes and Its Influence on Intraoperative Blood Loss and Postoperative Drainage.

Objective: To evaluate the efficacy of different doses of tranexamic acid plus traditional Chinese medicine (TCM) in hip arthroplasty in diabetic patients and the effect on intraoperative hemorrhage and postoperative drainage. Methods: One hundred patients admitted to our hospital from January 2019 to September 2021 were randomly divided into group B (n = 50) and group A (n = 50), and tranexamic acid was injected intravenously at a dose of 10 mg/kg and 20 mg/kg 30 min before skin incision, and then tranexamic acid 1.0 g was injected into the joint cavity through the drainage after incision closure, followed by 3 h of drainage clamping. The amount of blood loss, coagulation index, postoperative drainage, and incidence of venous thromboembolism (VTE) were compared between the groups. Results: Group A had significantly less total blood loss, dominant blood loss, and hidden blood loss than group B (P < 0.05). No significant difference in postoperative coagulation indexes and postoperative drainage flow was found between the two groups (P > 0.05). Serological examination results demonstrated no statistical difference in D-dimer (D-D) levels between the two groups. The absence of VTE in both groups was determined by imaging. Conclusion: Tranexamic acid is effective in reducing intraoperative hemorrhage in diabetic patients undergoing hip arthroplasty. The dose of 20 mg/kg outperforms 10 mg/kg in terms of clinical efficacy with a favorable safety profile, which can be applied according to the patient's actual condition.

Geographical origin traceability of traditional Chinese medicine Atractylodes macrocephala Koidz. by using multi-way fluorescence fingerprint and chemometric methods.

Atractylodes macrocephala Koidz. (AM) is an important plant of traditional Chinese medicine (TCM), and its status can be comparable with ginseng in China. The efficacy and quality of AM are closely related to the place of origin. Hence, we proposed a simple and fast strategy to classify AM from different geographical origins by using multi-way fluorescence fingerprint combined with chemometric methods. AM samples with different dilution levels have different fluorescence characteristics, resulting from different content of fluorescence components and chemical microenvironment. Therefore, AM samples were diluted 5-fold, 10-fold, and 20-fold with 40% ethanol aqueous solution to obtain excitation-emission matrix data, and multi-way (three-way and four-way) data arrays were constructed. And then, the fluorescence fingerprints of AM samples were characterized by three-way and four-way parallel factor analysis (PARAFAC). In addition, four pattern recognition methods were used to classify AM from different provinces. The results show that the four-way data array can provide more abundant information than three-way data arrays, so it is more conducive to sample classification. According to the results obtained from the analysis of four-way data array, the correct classification rate (CCR) of the cross-validation and prediction set obtained by partial least squares-discrimination analysis (PLS-DA) were 90.5% and 100%, respectively. To sum up, the proposed method can be regarded as a powerful, feasible, convenient, reliable, and universal classification tool for the classification of AM samples from different provinces and can be used as a promising method to realize the geographical origin traceability of other TCMs.

Novel dibenzofuran and biphenyl phytoalexins from Sorbus pohuashanensis suspension cell and their antimicrobial activities.

Two novel sulfur-containing dibenzofurans, sorbusins A (1) and B (2), two unprecedented biphenyl glycosides, 2'-hydroxyaucuparin 2'-O-ɑ-L-rhamnoside (3) and noraucuparin 5-O-ɑ-L-rhamnoside (4), and four known analogues (5-8), were isolated from Sorbus pohuashanensis suspension cell induced by yeast extract. Their structures were elucidated based on spectroscopic analyses and quantum calculation of 13C NMR data. Structurally, compound 1 possessed a rare naturally occurring benzothiazole moiety and represents the first example of thiazole fused dibenzofuran. A plausible biosynthetic pathway for the sulfur-containing dibenzofurans is proposed. These dibenzofuran and biphenyl phytoalexins were evaluated for their antimicrobial activities against pathogenic fungi and drug-resistant bacteria. Compound 7 exhibited significant antibacterial activity against methicinllin-resistant Staphylococcus aureus with an MIC value of 3.13 µg/mL.

A new fusicoccane diterpene and a new polyene from the plant endophytic fungus Talaromyces pinophilus and their antimicrobial activities.

A new fusicoccane diterpene, pinophicin A (1), and a new polyene, pinophol A (2), were isolated from the plant endophytic fungus Talaromyces pinophilus obtained from the aerial parts of Salvia miltiorrhiza. The structures and relative configurations of 1-2 were determined by the analysis of extensive spectroscopic data, chemical method, and comparison with known compounds. Compound 2 exhibited weak antibacterial activity against Bacterium paratyphosum B with an MIC value of 50 µg/mL.

[Inhibition of Copper Transporter-1 by Ammonium Tetrathiocarbolybdate in the Treatment of Pancreatic Cancer].

OBJECTIVE: To examine copper transporter 1 (CTR1) expression in pancreatic carcinoma cells, orthotopic xenograft pancreatic tumor model and clinical samples, and verify the effect of copper chelating agent ammonium tetrathiomolybdate (TM) regulate the expression of CTR1 in pancreatic carcinoma cells and the inhibition of pancreatic carcinoma. METHODS: The expressions of copper transporter CTR1 and antioxidant protein 1 (ATOX1) in 22 clinical pancreatic ductal carcinoma and paracancer tissues 0.5-1 cm away from the tumor were measured by immunohistochemistry (IHC). PANC-1 cells were used to construct 5 orthotopic xenograft pancreatic tumor of nude mice models. Pancreatic cancer tissues and corresponding normal pancreatic tissues were collected, and the expressions of CTR1 and ATOX1 were detected by IHC and compared with clinical tissues. The proliferation of pancreatic carcinoma cells PANC-1 treated with 10, 30, 50, 100 µmol/L TM for 24 h, 48 h, 72 h was measured by CCK8 assay. The migration abilities of PANC-1 cells treated with 50 µmol/L TM for 24 h, 48 h were detected by scratch test. The expressions of CTR1, vascular endothelial growth factor (VEGF) and CyclinD1 proteins in PANC-1 cells treated with 10, 30, 50, 100 µmol/L TM for 48 h were measured by Western blot. Then the subcutaneous tumor-bearing model of nude mice were established with PANC-1 cells, and the growth of tumor was observed after oral administration of 0.3 mg/d and 1.0 mg/d of TM, respectively. RESULTS: The immunohistochemical results indicated that 19 of the 22 clinical pancreatic ductal cancer tissues of carcinoma patients had high expression of CTR1, and the same high expression of CTR1 was found in the orthotopic transplanted tumor tissues of PANC-1 nude mice. The proliferation inhibition of PANC-1 cells increased with the concentration of TM increased and the treatment time prolonged. The expressions of intracellular CTR1, VEGF and CyclinD1 all decreased with the concentration of TM increased. The cell migration ability decreased after the PANC-1 cells treated with TM. The tumor growth of PANC-1 tumor-bearing nude mice was inhibited after different doses of TM were delivered. The reduction in tumor volume and weight was more pronounced in the high-dose TM group (P<0.05). CONCLUSION: The expression of CTR1 is abnormally elevated in pancreatic carcinoma, and treatment with copper chelating agent for this target may help to inhibit pancreatic carcinoma.

Two novel eremophylane acetophenone conjugates from Colletotrichum gloeosporioides, an endophytic fungus in Salvia miltiorrhiza.

Two novel eremophylane acetophenone conjugates, colletotricholides A (1) and B (2), were isolated from the solid fermentation cultures of an endophytic fungus Colletotrichum gloeosporioides XL1200 isolated from the aerial parts of Salvia miltiorrhiza. The chemical structures of 1-2 were characterized by extensive spectroscopic methods and single-crystal X-ray crystallography. Structurally, compounds 1-2 are two unusual eremophylane acetophenone conjugates originating from the hybrid pathways of polyketide synthase and sesquiterpene synthase. In addition, compounds 1-2 were inactive against tested pathogens.

(±)-Preisomide: A new alkaloid featuring a rare naturally occurring tetrahydro-2H-1,2-oxazin skeleton from an endophytic fungus Preussia isomera by using OSMAC strategy.

A new alkaloid, named (±)-preisomide (1), together with five known polyketides (2-6), were isolated from an endophytic fungus Preussia isomera in Panax notoginseng by using one strain-many compounds (OSMAC) strategy. Their structures were identified by extensive spectroscopic experiments and comparison with literature data. Structurally, compound 1 possessed a rare naturally occurring tetrahydro-2H-1,2-oxazin ring. Compound 6 displayed significant antibacterial activity against multidrug-resistant Enterococcus faecium, methicinllin-resistant Staphylococcus aureus and multidrug-resistant Enterococcus faecalis with an MIC value of 25 µg/mL, as well as moderate antifungal activity against Gibberella saubinetii with an MIC value of 50 µg/mL.

Emerones A-C: three novel merosesquiterpenoids with unprecedented skeletons from Emericella sp. XL029.

Three novel highly oxygenated α-pyrone merosesquiterpenoids, emerones A-C (1-3), have been obtained from the fungus, Emericella sp. XL029, which was isolated from the leaves of the traditional Chinese medicinal plant, Panax notoginseng. The structures and absolute configurations of 1-3 were determined by NMR experiments, X-ray diffraction analysis, and computational methods. Structurally, compound 1 possessed an unprecedented 5/7 bicyclic ring architecture, compound 2 had an unusual substituted 10-membered ring, and compound 3 had an undescribed norsesquiterpene skeleton. In addition, compounds 1 and 2 showed moderate activity towards several types of bacteria and fungi, exhibiting MIC values ranging from 12.5 to 50 µg mL-1, whereas compound 3 showed no antimicrobial activity.

Five new polyphenolic derivatives with antimicrobial activities from the root barks of Periploca sepium.

Five new polyphenolic derivatives, sepiumols A-E (1-5), were isolated from the root barks of Periploca sepium. Their structures were elucidated by interpretation of NMR spectroscopic and mass spectrometric data. Compounds 1, 3 and 5 were found to exhibit significant antifungal activity, particularly for 3 with the remarkable activity against Gibberella saubinetii and Alternaria longipes with MIC values of 1.56 and 3.13 µg/mL (ketoconazole: 0.78 µg/mL), respectively. In addition, compounds 1, 3 and 5 also displayed significant antibacterial activity against methicillin-resistant Staphylococcus aureu with MIC values of 12.50-25 µg/mL (ciprofloxacin: 0.78 µg/mL).

Sesquiterpenoids and diterpenes with antimicrobial activity from Leptosphaeria sp. XL026, an endophytic fungus in Panax notoginseng.

Two new sesquiterpenoids, leptosphins A (1) and B (2), and a new cyclopiane diterpene, leptosphin C (3), along with four known diterpenes (4-7) were isolated from the solid fermentation cultures of an endophytic fungus Leptosphaeria sp. XL026 isolated from the leaves of Panax notoginseng. Their structures were elucidated by extensive spectroscopic methods and single-crystal X-ray diffraction (data). Compound 1 represents the first sulfur-containing eremophilane sesquiterpene. Compounds 5 and 7 displayed medium antifungal activity against Rhizoctonia cerealis, as well as 6 against Verticillium dahliae Kleb with an MIC value of 12.5 µg/mL. Furthermore, compounds 2, 5, 6 and 7 showed medium antibacterial activity against Bacillus cereus with MIC values of 12.5-6.25 µg/mL, as well as 6 also against Pseudomonas aeruginosa with an MIC value of 12.5 µg/mL.

Three new hopane-type triterpenoids from the aerial part of Adiantum capillus-veneris and their antimicrobial activities.

Three new hopane-type triterpenoids (1-3), fern-7(8)-en-19α, 28-diol (1), pteron-14-ene-7α,19α,28-triol (2) and 3ß,4α,25-trihydroxyfilican (3), were isolated from the aerial parts of Adiantum capillus-veneris. Their structures were determined by NMR spectroscopic and mass spectrometric data. Compounds 2 and 3 exhibited remarkable antifungal activity against Helminthosporium maydis and Alternaria alternata with MIC values of 12.5-3.125 µg/mL, and compound 3 also against Verticillium dahliae Kleb with an MIC value of 3.125 µg/mL. In addition, compounds 1-3 also displayed weak antibacterial activity against Micrococcus lysodeikticus, Bacterium paratyphosum B and Pseudomonas aeruginosa with an MIC value of 100 µg/mL.

Two new triterpenoids with antimicrobial activity from the leaves and twigs of Orophea yunnanensis.

Two new triterpenoids, 3-O-(4',5'-dihydroxybenzoyl)-lup-20(29)-en (1) and 3-O-(6'-desmethysyringyl)-13α-methyl-27-norolean-14-en-3ß-ol (2), were isolated from the leaves and twigs of Orophea yunnanensis. Their structures were identified by extensive spectroscopic experiments (NMR and MS) and comparison with literature data. Compounds 1 and 2 exhibited the moderate inhibitory activity against Sclerotinia sclerotiorum, Ceratocystis fimbriata and Verticillium dahliae Kleb with MIC values from 50 to 25 µg/mL, and also displayed the weak activity selectively against tested bacteria strains with MIC values from 100 to 50 µg/mL.

Emericellins A and B: Two sesquiterpenoids with an unprecedented tricyclo[4,4,2,1]hendecane scaffold from the liquid cultures of endophytic fungus Emericella sp. XL 029.

Two novel sesquiterpenoids with an unprecedented tricyclo[4,4,2,1]hendecane scaffold, namely emericellins A (1) and B (2) representing a new skeleton, were isolated from the liquid cultures of an endophytic fungus Emericella sp. XL 029 associated with the leaves of Panax notoginseng. Their structures and relative configurations were characterized by extensive spectroscopic methods. Compounds 1-2 displayed moderate activities against three fungal strains (Verticillium dahliae Kleb, Helminthosporium maydis, and Botryosphaeria dothidea) and three bacterial strains (Bacillus subtilis, Bacillus cereus and Escherichia coli) with MIC values of 25-50 µg/mL.

Three new phenanthrenes with antimicrobial activities from the aerial parts of Juncus effusus.

Three new phenanthrenes (1-3), designated as 2-methoxy-1,6-dimethyl-5-vinyl-9, 10-dihydrophenanthren-7-ol, 1,6-dimethyl-4,5-dihydropyrene-2,7-diol and 1-(3,7- dihydroxy-2,8-dimethyl-9,10-dihydrophenanthren-1-yl)ethanone, were isolated from the aerial parts of Juncus effusus. Their structures were determined by extensive spectroscopic experiments (NMR and MS) and comparing with those related known compounds. The antifungal and antibacterial activities of 1-3 were evaluated. Compound 1 showed remarkable antifungal activities against six agricultural pathogenic fungi (Rhizoctonia solani, Verticillium dahliae Kleb, Sclerotinia sclerotiorum, Gibberella saubinetii, Bipolaris zeicola, and Phytophthora parasitica) with minimum inhibitory concentration (MIC) values ranging from 3.125 to 12.5 µg/mL, and also displayed significant antibacterial activities against two human pathogenic bacteria (Bacterium paratyphosum B and Micrococcus lysodeikticus) with MIC values of 12.5 and 25 µg/mL, respectively.

New alkenylated tetrahydropyran derivatives from the marine sediment-derived fungus Westerdykella dispersa and their bioactivities.

Six new alkenylated tetrahydropyran derivatives belonged to polyketides, designated as (12R,13R)-dihydroxylanomycinol (1), (12S,13S)-dihydroxylanomycinol (2), (12R,13S)-dihydroxylanomycinol (3) and (12S,13R)-dihydroxylanomycinol (4), (12S,13R)-N-acetyl-dihydroxylanomycin (5) and (12S,13S)-N-acetyl-dihydroxylanomycin (6), together with one related known compound lanomycinol (7) were isolated from the liquid cultures of Westerdykella dispersa obtained from the marine sediments. Their structures and absolute configurations were determined on the basis of extensive analysis of NMR spectra combined with comparison of NMR data to those of known compound, and computational method via calculation of the electronic circular dichroism (ECD). The anti-agricultural pathogenic fungal and antibacterial activities of all isolated compounds were evaluated. Compounds 1-7 exhibited moderate antifungal activities selectively against tested fungal strains with minimum inhibitory concentration (MIC) values ranging from 12.5 to 50µg/mL, and weak antibacterial activities selectively against tested antibacterial strains with MIC value at 100µg/mL.

Dietary flavonoid luteolin attenuates uropathogenic Escherichia. Coli invasion of the urinary bladder.

Uropathogenic Escherichia coli (UPEC), the primary uropathogen, adhere to and invade bladder epithelial cells (BECs) to establish a successful urinary tract infection (UTI). Emerging antibiotic resistance requires novel nonantibiotic strategies. Our previous study indicated that luteolin attenuated adhesive and invasive abilities as well as cytotoxicity of UPEC on T24 BECs through down-regulating UPEC virulence factors. The aims of this study were to investigate the possible function of the flavonoid luteolin and the mechanisms by which luteolin functions in UPEC-induced bladder infection. Firstly, obvious reduction of UPEC invasion but not adhesion were observed in luteolin-pretreated 5637 and T24 BECs sa well as mice bladder via colony counting. The luteolin-mediated suppression of UPEC invasion was linked to elevated levels of intracellular cAMP induced by inhibiting the activity of cAMP-phosphodiesterases (cAMP-PDEs), which resulting activation of protein kinase A, thereby negatively regulating Rac1-GTPase-mediated actin polymerization. Furthermore, p38 MAPK was primarily and ERK1/2 was partially involved in luteolin-mediated suppression of UPEC invasion and actin polymerization, as confirmed with chemical activators of p38 MAPK and ERK1/2. These data suggest that luteolin can protect bladder epithelial cells against UPEC invasion. Therefore, luteolin or luteolin-rich products as dietary supplement may be beneficial to control the UPEC-related bladder infections, and cAMP-PDEs may be a therapy target for UTIs treatment. © 2016 BioFactors, 42(6):674-685, 2016.

Dihydroberkleasmin A: a new eremophilane sesquiterpenoid from the fermentation broth of the plant endophytic fungus Pestalotiopsis photiniae.

Dihydroberkleasmin A (1), a new ester-substituted sesquiterpenoid related to the eremophilane class, together with the known compound berkleasmin C (2), were isolated from the fermentation broth of the plant endophytic fungus Pestalotiopsis photiniae. The structure of dihydroberkleasmin A (1) was elucidated by extensive spectroscopic analysis. The stereochemistry was assigned by comparison of the NMR spectroscopic data with those of berkleasmin A.