Screening and identification of antibacterial components in Artemisia argyi essential oil by TLC-direct bioautography combined with comprehensive 2D GC × GC-TOFMS.

One of the primary components that contribute to Artemisia argyi 's effectiveness is essential oil, which has an exceptional antibacterial effect that has been well documented. The actual cause of its antibacterial activity and associated mechanism, however, are still not completely understood. For the first time, comprehensive two-dimensional gas chromatography coupled with time-of-flight mass spectrometry (2D GC × GC-TOFMS) and thin-layer chromatography-direct bioautography (TLC-DB) were employed to investigate its antibacterial components. The antibacterial properties of A. argyi essential oil were investigated, and the antibacterial activity of six compounds was evaluated, using Staphylococcus aureus (S. aureus) and Escherichia coli (E. coil) as test microorganisms. TLC-direct bioautography was used to screen two bioactive clusters. Following 2D GC × GC-TOFMS identification of bioactive clusters, six compounds were evaluated for in vitro antibacterial activity verification. All the components tested displayed antibacterial action. Results showed that α-terpineol and eugenol had high potent antibacterial activity (MIC＜0.62 mg/mL, IC50＜2.00 mg/mL). For complex essential oils from traditional Chinese medicine, this method is efficient for quick screening and identifying antibacterial compounds.

Review of treatment options for a multidrug-resistant fungus: Candida auris.

Candida auris is a widely distributed, highly lethal, multidrug-resistant fungal pathogen. It was first identified in 2009 when it was isolated from fluid drained from the external ear canal of a patient in Japan. Since then, it has caused infectious outbreaks in over 45 countries, with mortality rates approaching 60%. Drug resistance is common in this species, with a large proportion of isolates displaying fluconazole resistance and nearly half are resistant to two or more antifungal drugs. In this review, we describe the drug resistance mechanism of C. auris and potential small-molecule drugs for treating C. auris infection. Among these antifungal agents, rezafungin was approved by the US Food and Drug Administration (FDA) for the treatment of candidemia and invasive candidiasis on March 22, 2023. Ibrexafungerp and fosmanogepix have entered phase III clinical trials.

Metagenomic Analysis of Gut Microbiome in Gout Patients with Different Chinese Traditional Medicine Treatments.

Introduction: Changes in eating habits have made gout a metabolic disease of increasing concern. Previous studies have indicated that there are significant differences in species composition and abundance of gut microbiome in gout patients compared with average. Considering that traditional Chinese medicine has a momentous effect in treating gout, the research study aimed to explore the differences of genomic and metabolomics of gut microbiome before and after traditional Chinese medicine treatment in patients with gout. Method: 30 patients with gout and 29 matched controls were recruited of which 16 patients took H treatment and 14 patients took T treatment. Stools were collected twice for patients before and after treatment and only once for controls. A total of 89 samples were annotated with metagenomic species and functions, and the enrichment analysis of differential genes and KO pathway was carried out. Result: The results showed a decrease in the diversity of gut microbiome in gout patients and the gene abundance and metabolomics had great differences among study groups. The number of bacterial genera also had significant differences among treatment groups. Moreover, among different groups, the regulation of different species was variously correlated. The correlation between species and clinical laboratory indicators in the rising group was stronger than that in the decreasing group and the upregulation of some strain was related to the content of urea nitrogen. Conclusion: After the traditional Chinese medicine treatment, the glutathione pathway was significantly enriched and some pathogenic bacteria were significantly inhibited. The study suggests that traditional Chinese medicine treatment may exert its therapeutic effect by inhibiting relevant pathways.

Fluorescent Probes and Mass Spectrometry-Based Methods to Quantify Thiols in Biological Systems.

Significance: Fluorescent probes and mass spectrometry are the two most popular and complementary methods to quantify thiols in biological systems. In this review, we focus on the widely used and commercially available methods to detect and quantify thiols in living cells and the general approaches applied in mass spectrometry-based thiol quantification. We hope that this review can serve as a general guide for redox biologists who are interested in thiol species. Sulfur, one of the most important elements in living systems, contributes to every aspect of physiology and pathology. Thiols, including cysteine, homocysteine, glutathione, hydrogen sulfide, and hydropersulfides, are the main players in the redox biology system. Therefore, quantifying these thiol species in biological systems is one of the important steps to understand their roles in biology. Recent Advances: Fluorescent probes and mass spectrometry-based methods have been developed to detect and/or quantify thiols in biological systems. Mass spectrometry-based methods have been the gold standard for metabolite quantification in cells. Fluorescent probes can directly detect or quantify thiol species in living cells with spatial and temporal resolutions. Additionally, organelle-specific fluorescent probes have been widely developed. These two methods are complementary to each other. Critical Issues: Reliable quantification of thiol species using fluorescent probes remains challenging. Future Directions: When developing fluorescent probes, we suggest using both the fluorescent probes and mass spectrometry-based thiol quantification methods to cross-check the results. In addition, we call on chemical biologists to move beyond qualitative probes and focus on probes that can provide quantitative results in live cells. These quantitative measurements based on fluorescent probes should be validated with mass spectrometry-based methods. More importantly, chemical biologists should make their probes accessible to the biology end users. Regarding mass spectrometry-based methods, quantification of the derivatized thiol specifies should fit into the general metabolomics workflow. Antioxid. Redox Signal. 36, 354-365.

Comparative pharmacokinetic study of six lignans in normal and diabetic rats after oral administration of Saururus chinensis extract by LC-MS/MS.

Saururus chinensis (SC) possesses significant anti-diabetic activity and lignans were its major bioactive compounds. In this study, a rapid and sensitive ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was established for simultaneous quantification of six lignans, namely (-)-(7R,8R)-machilin D (1), verrucesin (2), rel-(7S,8S,7'R,8'R)-3,3',4,4',5,5'-hexamethoxy-7.O.7',8.8'-lignan (3), manassantin A (4), manassantin B (5), and saucerneol F (6) in rat's plasma. It was validated with acceptable linearity (r ≥ 0.9922), accuracy (80.42-95.17%), precision (RSD ≤ 12.08%), and extraction recovery (80.36-93.45%). The method was successfully applied to the comparative pharmacokinetic study of the six lignans in normal and diabetic rats after oral administration of SC extract. Results showed that the areas under the plasma concentration-time curve (AUC0 → t and AUC0 → ∞ ) of (-)-(7R,8R)-machilin D, rel-(7S,8S,7'R,8'R)-3,3',4,4',5,5'-hexamethoxy-7.O.7',8.8'-lignan, manassantin B, and saucerneol F in diabetic rats were significantly increased, and the plasma clearance (CL) of (-)-(7R,8R)-machilin D in diabetic rats was significantly decreased. However, the AUC0 → t and AUC0 → ∞ of verrucesin were significantly decreased, and its CL was significantly increased in diabetic rats compared with those in normal rats. These results indicated that there were remarkable differences in the pharmacokinetic parameters between the normal and diabetic rats. The pharmacokinetic studies might be beneficial for the clinical use of SC as hypoglycemic agent.

Acute and subacute toxicity evaluation of Houttuynia cordata ethanol extract and plasma metabolic profiling analysis in both male and female rats.

Houttuynia cordata has been used as a traditional medicine for more than 1500 years. It has aroused wide public concern about its safety in the past few years, for it contains various aristolactams. However, the safety of H. cordata extract remains unclear. In the present study, single dose (2000 mg/kg) and subacute (250, 500, and 1000 mg/kg/day for 28 days) oral toxicity studies of the 95% ethanol extract of H. cordata (HCE) were performed in both male and female Sprague-Dawley (SD) rats. Hematological, biochemical, histopathological parameters, and plasma metabolic profiling were assessed. The single-dose toxicity of HCE was more than 2000 mg/kg. The subacute toxicity results showed that no significant adverse effect of HCE was observed at 250 mg/kg/day. However, five rats died in 500 and 1000 mg/kg/day groups and exhibited toxicities to liver and kidney. Plasma metabolic profiling analysis suggested that a number of metabolic disturbances were induced by oral administration of HCE, focusing on energy metabolism, amino acid metabolism, and lipids metabolism. Moreover, it appeared that male rats were more susceptible to the toxic effects of HCE than female rats. Therefore, in this preliminary study, oral administration of HCE 250 mg/kg/day can be regarded as the no observed adverse effect level in rats over 28 days. However, long-term use of HCE with large doses exhibited some hepatotoxicity and nephrotoxicity in rats.

Metaproteomics reveals the molecular mechanism underlying bloom maintenance of a marine dinoflagellate under low ambient CO2 and inorganic nutrients.

Dinoflagellate blooming periods are paradoxically characterized by high biomass growth rate and low ambient dissolved CO2 and inorganic nutrients, however, the underlying mechanisms linking cell growth and nutrient acquisition are poorly understood. Here, we compared metaproteomes of non-bloom, mid-blooming and late-blooming cells of a marine dinoflagellate Prorocentrum donghaiense. Cell division, metabolism of carbon, nitrogen, phosphorus, lipid, porphyrin and chlorophyll were more active in blooming cells than in non-bloom cells. Up-regulation of carbonic anhydrase, ribulose-1,5-bisphosphate carboxylase/oxygenase II, and C4-cycle proteins enhanced CO2 assimilation of P. donghaiense. Proteins participating in external organic nutrient acquisition and conversion, such as transporters for fatty acids, peptides and amino acids, external- and internal-phosphomonoester hydrolase, and diverse peptidases and amino acid transaminases, exhibited higher expression in blooming cells relative to non-bloom cells. Interestingly, dissolved organic nitrogen (DON) such as urea and aspartate significantly down-regulated expression and activity of carbon assimilation proteins except for RuBisCO form II, suggesting that DON provided sufficient carbon source which reduced the need to concentrate internal CO2. This study demonstrates that coupling of efficient CO2 assimilation with DON utilization are essential for bloom maintenance of P. donghaiense, and future efforts should be devoted to dissolved organic nutrients for prevention and management of dinoflagelllate blooms.

Inulin supplementation ameliorates hyperuricemia and modulates gut microbiota in Uox-knockout mice.

PURPOSE: Inulin is a type of fermentable dietary fiber, which is non-digestible, and can improve metabolic function by modulating intestinal microbiota. This study aimed to evaluate the role of inulin in hyperuricemia and microbial composition of the gut microbiota in a mouse model of hyperuricemia established through knockout of Uox (urate oxidase) gene. METHODS: KO (Uox-knockout) and WT (wild-type) mice were given inulin or saline by gavage for 7 weeks. The effect of inulin to combat hyperuricemia was determined by assessing the changes in serum UA (uric acid) levels, inflammatory parameters, epithelial barrier integrity, fecal microbiota alterations, and SCFA (short-chain fatty acid) concentrations in KO mice. RESULTS: Inulin supplementation can effectively alleviate hyperuricemia, increase the expressions of ABCG2 in intestine, and downregulate expression and activity of hepatic XOD (xanthine oxidase) in KO mice. It was revealed that the levels of inflammatory cytokines and the LPS (lipopolysaccharide) were remarkably higher in the KO group than those in the WT group, indicating systemic inflammation of hyperuricemic mice, but inulin treatment ameliorated inflammation in KO mice. Besides, inulin treatment repaired the intestinal epithelial barrier as evidenced by increased levels of intestinal TJ (tight junction) proteins [ZO-1 (zonula occludens-1) and occluding] in KO mice. Moreover, serum levels of uremic toxins, including IS (indoxyl sulfate) and PCS (p-cresol sulfate), were reduced in inulin-treated KO mice. Further investigation unveiled that inulin supplementation enhanced microbial diversity and raised the relative abundance of beneficial bacteria, involving SCFAs-producing bacteria (e.g., Akkermansia and Ruminococcus). Additionally, inulin treatment increased the production of gut microbiota-derived SCFAs (acetate, propionate and butyrate concentrations) in KO mice, which was positively correlated with the effectiveness of hyperuricemia relief. CONCLUSIONS: Our findings showed that inulin may be a promising therapeutic candidate for the treatment of hyperuricemia. Moreover, alleviation of hyperuricemia by inulin supplementation was, at least, partially conciliated by modulation of gut microbiota and its metabolites.

Characterization, evaluation of nutritional parameters of Radix isatidis protein and its antioxidant activity in D-galactose induced ageing mice.

BACKGROUND: Radix isatidis (Isatis indigotica Fort.) is an ancient medicinal herb, which has been applied to the prevention and treatment of influenza virus since ancient times. In recent years, the antioxidant activity of Radix isatidis has been widely concerned by researchers. Our previous studies have shown that Radix isatidis protein (RIP) has good antioxidant activity in vitro. In this study, the composition of the protein was characterized and its antioxidant activity in vivo was evaluated. METHODS: The model of oxidative damage in mice was established by subcutaneous injection of D-galactose for 7 weeks. Commercially available kits were used to determine the content of protein and several oxidation indexes in different tissues of mice. The tissue samples were stained with hematoxylin and eosin (H&E) and the pathological changes were observed by optical microscope. The molecular weight of RIP was analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The amino acid composition of RIP was determined by a non-derivative method developed by our research group. RESULTS: RIP significantly increased the activities of antioxidant enzymes such as SOD, CAT, GSH-Px and total antioxidant capability (TAOC) but decreased the MDA level in the serum, kidney and liver. H&E stained sections of liver and kidney revealed D-galactose could cause serious injury and RIP could substantially attenuate the injury. The analysis of SDS-PAGE showed that four bands with molecular weights of 19.2 kDa, 21.5 kDa, 24.8 kDa and 40.0 kDa were the main protein components of RIP. CONCLUSIONS: The results suggested that RIP had excellent antioxidant activity, which could be explored as a health-care product to retard aging and a good source of protein nutrition for human consumption.

Trapa natans pericarp extract ameliorates hyperglycemia and hyperlipidemia in type 2 diabetic mice

Abstract The pericarp of Trapa natans L., an annual aquatic floating herb belonging to Lythraceae family, is used as a folk medicine in China. In this study, extracts of Trapa natans pericarp were tested both in vitro and in vivo through a high-fat diet with a single medium dosage streptozotocin injection induced type 2 diabetic mice. Different solvent extracts of Trapa natans pericarp showed α-amylase and α-glucosidase inhibitory activity. After four weeks administration, the ethyl acetate extract of Trapa natans pericarp (50 and 100 mg/kg b.w.) reduced fasting blood glucose level, ameliorated oral glucose tolerance and insulin resistance, improved serum lipids alterations in type 2 diabetic mice as well. Additionally, ethyl acetate extract significantly elevated the insulin receptor substrate 1 and Akt serine/threonine kinase phosphorylation compared to diabetic group. HPLC-MS and HPLC-DAD analysis showed that the ethyl acetate extract was rich in hydrolysable tannins. Results support the notion that Trapa natans pericarp extract has a potential hypoglycemic activity.

Differences in Serum Amino Acid Phenotypes Among Patients with Diabetic Nephropathy, Hypertensive Nephropathy, and Chronic Nephritis.

BACKGROUND We assessed levels of circulating amino acids in different etiologies of chronic kidney disease (CKD) and the association of amino acids with risk factors of CKD progression. MATERIAL AND METHODS High-performance liquid chromatography-based analysis was used to determine amino acid profiles in patients with diabetic nephropathy (DN, n=20), hypertensive nephropathy (HN, n=26), and chronic nephritis (CN, n=33), and in healthy controls (HC, n=25). RESULTS All 3 types of CKD patients displayed decreased serum levels of serine, glycine, GABA, and tryptophan compared with healthy controls. Moreover, serine and tryptophan were positively correlated with glucose in DN cohorts. Total cholesterol was positively correlated with tryptophan levels in the DN cohort and negatively correlated with serine levels in the CN cohort. In the HN cohort, glycine was negatively correlated with triglyceride levels, and systolic blood pressure (SBP) was negatively correlated with GABA levels. CONCLUSIONS Patients with different etiologies of CKD have significantly different amino acids profiles, and this indicates specific supplementary nutritional needs in CKD patients.

Chemical constituents from the stems of Securidaca inappendiculata Hassk.

Three new neolignan glycosides, (7R,8S)-4-hydroxy-3,3'-dimethoxy-8,4'-oxyneoligna-7,9,9'-triol-4-O-ß-d-glucopyranosyl-(1 → 4)-ß-D-glucopyranoside (1), (7R,8S)-4-hydroxy-3,5'-dimethoxy-4',7-epoxy-8,3'-neoligna-9,9'-diol-9'-O-ß-d-glucopyranosyl-4-O-[ß-d-glucopyranosyl-(1 → 4)]-ß-D-glucopyranoside (2), and (7R,8S)-4-hydroxy-3,5,5'-trimethoxy-4',7-epoxy-8,3'-neoligna-9,9'-diol-9'-O-ß-d-glucopyranosyl-4-O-[ß-d-glucopyranosyl-(1 → 4)]-ß-D-glucopyranoside (3), one new phenolic glycoside, securiphenoside B (4) and two new hemiterpene glycosides, securiterpenoside E-F (5-6) were isolated from the stems of Securidaca inappendiculata Hassk. Their structures were elucidated on the basis of 1D and 2D NMR, HRESIMS, CD and chemical evidence. Furthermore, compound 2 showed moderate hepatoprotective activity compared with bicyclol in vitro.

AhlX, an N-acylhomoserine Lactonase with Unique Properties.

N-Acylhomoserine lactonase degrades the lactone ring of N-acylhomoserine lactones (AHLs) and has been widely suggested as a promising candidate for use in bacterial disease control. While a number of AHL lactonases have been characterized, none of them has been developed as a commercially available enzymatic product for in vitro AHL quenching due to their low stability. In this study, a highly stable AHL lactonase (AhlX) was identified and isolated from the marine bacterium Salinicola salaria MCCC1A01339. AhlX is encoded by a 768-bp gene and has a predicted molecular mass of 29 kDa. The enzyme retained approximately 97% activity after incubating at 25 °C for 12 days and ~100% activity after incubating at 60 °C for 2 h. Furthermore, AhlX exhibited a high salt tolerance, retaining approximately 60% of its activity observed in the presence of 25% NaCl. In addition, an AhlX powder made by an industrial spray-drying process attenuated Erwinia carotovora infection. These results suggest that AhlX has great potential for use as an in vitro preventive and therapeutic agent for bacterial diseases.

Eight new cytotoxic annonaceous acetogenins from the seeds of Annona squamosa.

Eight new annonaceous acetogenins, squamotin A-D (1-4), annosquatin IV-V (5 and 6), muricin O (7) and squamosten B (8), together with four known ones (9-12) were isolated from the seeds of Annona squamosa. Their structures were elucidated by chemical methods and spectral data. The inhibitory activities of compound 1-9 against three multidrug resistance cell lines were evaluated. All tested compounds showed strong cytotoxicity.

Fusariumins C and D, two novel antimicrobial agents from Fusarium oxysporum ZZP-R1 symbiotic on Rumex madaio Makino.

Bioassay-guided fractionation of the crude extract of fermentation broth of one symbiotic strain Fusarium oxysporum ZZP-R1 derived from coastal plant Rumex madaio Makino, one traditional Chinese medicine used as a treatment of inflammation and toxication, yielded two novel compounds, fusariumins C (1) and D (2). Chemical structures of 1 and 2 were respectively determined as one meroterpene with cyclohexanone moiety and a sesquiterpene ester with a conjugated triene and an unusual oxetene ring by a combination of spectroscopic methods, including 1D and 2D NMR, mass spectrometry, and optical rotation analysis, as well as by comparison with literature data. Bioassay results indicated that compound 1 displayed potent activity against Staphyloccocus aureus with an MIC value of 6.25 µM, and compound 2 had a moderate inhibitory effect on S. aureus with an MIC value of 25.0 µM. It was the first report that phytochemical investigation of Fusarium strain from R. madaio Makino led to isolation of new antimicrobial agents.

Quorum sensing inhibitors from marine bacteria Oceanobacillus sp. XC22919.

In this study, three active compounds isolated from Oceanobacillus sp. XC22919 were identified as 2-methyl-N-(2'-phenylethyl) butyramide (1), 3-methyl-N-(2'-phenylethyl)-butyramide (2) and benzyl benzoate (3), and were first reported to exhibit the apparent quorum sensing inhibitory activities against C. violaceum 026 and P. aeruginosa. Compounds 1-3 inhibited violacein production of C. violaceum 026 by 10.5-55.7, 11.2-55.7, and 27.2%-95.7%, respectively, and inhibited pyocyanin production of P. aeruginosa by 1.7-50.8, 39.1-90.7, and 57.2%-98.7%, respectively. The azocasein-degrading proteolytic rates of P. aeruginosa were observed by 13.4-31.5, 13.4-28.8, and 11.3%-21.1%, respectively. With respect to elastase, the range of inhibition of activity of compounds 1-3 was 2.1-30.3, 4.2-18.2, and 8.9%-15.7%, respectively. Compounds 1 and 3 also showed a concentration-dependent attenuation in biofilm formation, with the maximum of 50.6% inhibition, and 37.7% inhibition at 100 µg/mL, respectively.

Matrix Solid-Phase Dispersion Combined with HPLC-DAD for Simultaneous Determination of Nine Lignans in Saururus chinensis.

A simple and rapid method, based on matrix solid-phase dispersion (MSPD) and high-performance liquid chromatography (HPLC) was developed for simultaneous determination of nine lignans, including (-)-(7R,8R)-machilin D (Wang, C., Wang, P., Chen, X., Wang, W., Jin, Y.; Saururus chinensis (Lour.) Baill blocks enterovirus 71 infection by hijacking MEK1-ERK signaling pathway; Antiviral Research, (2015); 119:47-56), dihydroguaiaretic acid (Quan, Z., Lee, Y.J., Yang, J.H., Lu,Y., Li,Y., Lee,Y.K., et al.; Ethanol extracts of Saururus chinensis suppress ovalbumin-sensitization airway inflammation; Journal of Ethnopharmacology, (2010); 132:143-149.), sauchinone (Zhuang, T., Liang, J.Y., Sun, J.B., Wu, Y., Huang, L.R., Qu, W.; Secondary metabolites from Saururus chinensis and their chemotaxonomic significance; Biochemical Systematics and Ecology, (2014); 56:95-98.), rel-(7S,8S,7'R,8'R)-3,3',4,4',5,5'-hexamethoxy-7.O.7',8.8'-lignan (Tsai, W.J., Shen, C.C., Tsai, T.H., Lin, L.C.; Lignans from the aerial parts of Saururus chinensis: isolation, structural characterization, and their effects on platelet aggregation; Journal of Natural Products, (2014); 77:125-131), licarin A (Cui, H., Xu, B., Wu, T., Xu, J., Yuan, Y., Gu, Q.; Potential antiviral lignans from the roots of Saururus chinensis with activity against Epstein-Barr virus lytic replication; Journal of Natural Products, (2014); 77:100-110.), manassantin A (Lu, Y., Piao, D., Zhang, H., Li, X., Chao, G.H., Park, S.J., et al.; Saucerneol F inhibits tumor necrosis factor-alpha and IL-6 production by suppressing Fyn-mediated pathways in FcepsilonRI-mediated mast cells; Food and Chemical Toxicology, (2013); 59:696-702.), saurucinol I (Kwon, O.E., Lee, H.S., Lee, S.W., Chung, M.Y., Bae, K.H., Rho, M.C., et al.; Manassantin A and B isolated from Saururus chinensis inhibit TNF-α-induced cell adhesion molecule expression of human umbilical vein endothelial cells; Archives of Pharmacal Research, (2005); 28:55-60.), manassantin B (Hwang, B.Y., Lee, J.H., Jung, H.S., Kim, K.S., Nam, J.B., Hong, Y.S., et al.; Sauchinone, a lignan from Saururus chinensis, suppresses iNOS expression through the inhibition of transactivation activity of RelA of NF-κB; Planta Medica, (2003); 69:1096-01.) and licarin B (Hwang, B.Y., Lee, J.H., Nam, J.B., Hong, Y.S., Lee, J.J.; Lignans from Saururus chinensis inhibiting the transcription factor NF-κB; Phytochemistry, (2003); 64:765-771.) in Saururus chinensis. The parameters of MSPD were optimized to be that 0.2 g of sample, blended with 0.4 g silica gel, and eluted with 5 mL of methanol. The separation was carried out on a C18 column with acidified aqueous acetonitrile gradients. The established method was fully validated in terms of linearity (r2 ≥ 0.9994), sensitivity, precision (RSD ≤ 3.18%), repeatability (RSD ≤ 3.02%) as well as recovery (93.49-103.52%), and subsequently applied to six samples of S. chinensis from different areas. The MSPD extraction promoted higher extraction yields of nine lignans, lower solvent sample and solvent consumption.

Metabolic profiling investigation of Fritillaria thunbergii Miq. by gas chromatography-mass spectrometry.

Thunberg fritillary bulb (the dry bulbs of Fritillaria thunbergii Miq.), a traditional Chinese Medicine, is widely applied as an expectorant and antitussive. In this investigation, the primary metabolites of bulbs, flowers, leaves, and stems of F. thunbergii were analyzed by gas chromatography-mass spectrometry. Principal component analysis, partial least squares-discriminate analysis, orthogonal projection to latent structures-discriminate analysis, and heat map analysis showed that there were dissimilar metabolites, and a negative correlation between amino acids and saccharides in different analytes. Furthermore, carbodiimide, tryptophan, glucose-6-phosphate, xylose, 2-piperidinecarboxylic acid, monoamidomalonic acid, phenylalanine, and histidine were found to play an important role in the plant metabolism net of F. thunbergii.

Simultaneous determination of four amides in Saururus chinensis by matrix solid phase dispersion and high-performance liquid chromatography method.

A rapid and simple analytical method was established for the determination of four amides (N-p-trans-coumaroyltyramine, aristolactam AⅡ, sauristolactam and aristolactam BⅡ) in Saururus chinensis by matrix solid phase dispersion (MSPD) and high-performance liquid chromatography-diode array detector (HPLC-DAD). In the optimized MSPD, 0.2 g S. chinensis powder was blended with 0.4 g silica gel, and 5 mL methanol was selected as elution solvent. The MSPD extraction achieved higher extraction recovery of four amides, and required less sample, solvent and preparation time, comparing with the conventional methods (Soxhlet and ultrasonic extraction). The assay was performed on a TSK gel ODS-100Z column (4.6 mm × 250 mm, 5 µm) at 30 °C. Acetonitrile and 0.4% acetic acid aqueous solution was used as mobile phase by gradient elution at the flow rate of 1.0 mL/min. The detection wavelength was 280 nm. All the analytes showed good linear regression (R2 ≥ 0.9998) within the concentration ranges. The validated method showed good precision and stability with relative standard deviations (RSDs) ≤ 3.18%. The recoveries were in the range of 96.57-99.65%, with RSDs less than 2.74%.

[Protective effect of Ligustri Lucidi Ait Polysaccharide against lipopolysaccharide-induced inflammatory injury of Sertoli cells in rats].

OBJECTIVE: To explore the effect of Ligustri Lucidi Ait Polysaccharide (LLP) on lipopolysaccharide (LPS)-induced inflammatory injury of Sertoli cells. METHODS: Rat Sertoli cells were isolated and cultured in vitro and then divided into five groups, blank control, LPS, LPS + low-dose LLP, LPS + medium-dose LLP, and LPS + high-dose LLP. After 48 hours of treatment, the proliferation of the cells was detected by CCK-8, their apoptosis determined by FMC, and the levels of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px) activity and malondialdehyde (MDA) in the supernatant of the cell culture medium measured by ultraviolet spectrophotometry. The contents of IL-1α, IL-6 and TGF-ß in the culture medium were detected by ELISA before and after removal of LPS. RESULTS: The proliferation of the cells showed statistically significant differences among different groups (F = 153.93, P < 0.01), markedly reduced in the LPS group as compared with the blank control (P < 0.01), but remarkably increased in the high- and medium-dose LLP groups in comparison with the LPS group (both P < 0.01), and so did the apoptosis of the cells (F = 64.06, P < 0.01), significantly increased in the LPS group as compared with the blank control (P < 0.05), but markedly decreased in the high- and medium-dose LLP groups in comparison with the LPS group (both P < 0.01). Statistically significant differences were also observed among different groups in the levels of SOD (F = 56.07, P < 0.01), CAT (F = 41.57, P < 0.01), GSH-Px activity (F = 238.46, P < 0.01), and MDA (F = 285.31, P < 0.01), with decreased SOD, CAT and GSH-Px activity (P < 0.01) and increased MDA (P < 0.01) in the LPS group as compared with the control, but elevated SOD and CAT in the high- and medium-dose LLP groups and increased GSH-Px activity and decreased MDA concentration in all the three LLP groups in comparison with the LPS group (P < 0.01). Before the removal of LPS, the contents of IL-1α, IL-6 and TGF-ß in the culture medium were markedly higher in the LPS than in the control group (all P < 0.01), that of IL-1α was increased significantly in the high- and medium-dose LLP groups (P < 0.01 and P < 0.05) while those of IL-6 and TGF-ß showed no statistically significant differences in the three LPS groups as compared with the LLP group (P > 0.05). After the removal of LPS, the contents of IL-1α and IL-6 were remarkably reduced (t = 25.26 and 61.43, P < 0.01) and that of TGF-ß increased (t = －18.16, P < 0.01), even more significantly in the LLP+LPS groups (P < 0.01). CONCLUSIONS: Ligustri Lucidi Ait Polysaccharide plays a protective role in LPS-induced inflammatory injury of Sertoli cells by reducing cell apoptosis and regulating the contents of IL-1α, IL-6 and TGF-ß from Sertoli cells in inflammation.