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冬虫夏草的人工繁殖与规模种植条件 药用价值的开发

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Analysis of sterols and fatty acids in natural and cultured Cordyceps by one-step derivatization followed with gas chromatography–mass spectrometry 人工培植的虫草与自然野外生长的虫草营养成分含量的区别 Pharmaceutical and Biomedical Analysis
Ten free fatty acids namely lauric acid, myristic acid, pentadecanoic acid, palmitoleic acid, palmitic acid, linoleic acid, oleic acid, stearic acid, docosanoic acid and lignoceric acid and four free sterols including ergosterol, cholesterol, campesterol and β-sitosterol in natural (wild) Cordyceps sinensis, Cordyceps liangshanensis and Cordyceps gunnii, as well as cultured C. sinensis and Cordyceps militaris were first determined using pressurized liquid extraction (PLE), trimethylsilyl (TMS) derivatization and GC–MS analysis. The conditions such as the amount of reagent, temperature and time for TMS derivatization of analytes were optimized. Under the optimum conditions, all calibration curves showed good linearity within the tested ranges. The intra- and inter-day variations for 14 investigated compounds were less than 3.4% and 5.2%, respectively. The results showed that palmitic acid, linoleic acid, oleic acid, stearic acid and ergosterol are main components in natural and cultured Cordyceps which could be discriminated by hierarchical clustering analysis based on the contents of 14 investigated compounds or the 4 fatty acids, where the contents of palmitic acid and oleic acid in natural Cordyceps are significantly higher than those in the cultured ones.

Clinical Application of Cordyceps sinensis on Immunosuppressive Therapy in Renal Transplantation Transplantation Proceedings

虫草临床应用价值的开发
We sought to explore the adjunctive effects of Cordyceps sinensis (CS) in clinical renal transplantation.

Materials and Methods

Patients (n = 202) were divided randomly by lottery into a treatment (n = 93) and a control group (n = 109). Patients in the treatment group were treated with CS 1.0 g 3 times a day in addition to the immunosuppressive regimen given to the control group. We compared patient and graft survivals, incidence, time and severity of acute rejection episodes, chronic allograft nephropathy (CAN), hepatotoxicity and nephrotoxicity, biochemistry parameters including indicators of liver and kidney functions, fats, proteinuria, dosages, and whole blood concentrations of cyclosporine (CsA).

Results

Patient and graft survival rates, serum creatinine (SCr), and blood urea nitrogen (BUN) were not significantly different between the 2 groups (P > .05). Serum uric acid (UA) and 24-hour urinary total

protein (24-hour UTP) were significantly lower in the treatment group than in the control group (P < .05). The incidences (11.83% vs 15.60%) and times to acute renal allograft rejection (23.48 ± 7.22 vs 22.27 ± 8.03 days posttransplantation) were not significantly different between the treated and control groups (P > .05). Patients receiving thymoglobulin antirejection therapy (3 cases) were fewer in the heated versus control group (13 cases; P = .014). The incidences of hepatotoxicity and nephrotoxicity in the treated group were 12.90% and 19.35%, significantly lower than 24.77% and 33.94% in the control group, respectively (P < .05). At 2 to 6 months posttransplantation, the CsA dosages in the treated group were significantly lower than those in the control group (P < .05). The whole blood trough CsA concentrations in the treated group were significantly lower than those in the control group at 3 to 6 months posttransplantation (P < .05). The decreasing trends of the 2 aforementioned parameters in the treatment group were approximately linear among treated subjects compared with approximately quadratic in the control group (P < .05). The incidence of CAN in the treated group was 7.53%, which was significantly lower than 18.35% in the control group (P = .024). The 24-hour UTP level in CAN patients within the treated group was significantly lower than the control group after transplantation (P = .045). The differences in total bilirubin, SCr, serum UA, and total cholesterol levels among otherwise normal patients in the treated group were significantly lower than those among the control group (P < .05).

Conclusions

The use of CS may allow decreased dosages and concentrations of CsA causing fewer side effects without an increased risk of acute rejection. In addition, CS with reduced dose CsA may decrease proteinuria and retard CAN progression.

Food Chemistry Acidic degradation and enhanced antioxidant activities of exopolysaccharides from Cordyceps sinensis mycelial culture
An exopolysaccharide (EPS) fraction isolated from mycelial culture of a Cordyceps sinensis fungus, designated EPS-1 with an average molecular weight (MW) of 38 kDa, was hydrolysed in dilute sulphuric acid solution at pH 1 and 90 ° yielding two major MW fractions, 3.0 kDa and 30 kDa, respectively. C, While the proportion of lower MW fraction increased with the hydrolysis period (18% in 0.5 h and 92% in 10 h), the polydispersity (Mw/Mn) of EPS decreased steadily (from 1.47 initially to 1.10 in 10 h). The IR

spectra of hydrolysed EPS fractions showed changes only in the C–O–C and C–O–H band peaks from that of EPS-1. These results suggest that the hydrolysis of EPS in the acidic solution caused the EPS degradation mainly by cleaving the glycosidic linkage but no change in the primary molecular structure. The hydrolysed EPS fractions had much higher (30–80%) antioxidant and radical-scavenging activities.

Cordysinocan, a polysaccharide isolated from cultured Cordyceps, activates immune responses in cultured T-lymphocytes and macrophages: Signaling cascade and induction of cytokines Ethnopharmacology
Cordyceps sinensis, a well-known traditional Chinese medicine, possesses activities in anti-tumor, anti-oxidation and stimulating the immune response; however, the identity of active component(s) is not determined. A strain of Cordyceps sinensis, namely UST 2000, has been isolated. By using activity-guided purification, a novel polysaccharide of molecular weight 82 kDa was isolated from the

conditioned medium of cultured Cordyceps. The isolated exo-polysaccharide, namely cordysinocan, contains glucose, mannose, galactose in a ratio of 2.4:2:1. In cultured T-lymphocytes, application of cordysinocan induced the cell proliferation and the secretion of interleukin-2, interleukin-6 and interleukin-8. In addition, the phosphorylation of extracellular signal-regulated kinases (ERK) was induced transiently by the treatment of cordysinocan. Moreover, application of cordysinocan in cultured macrophages increased the phagocytosis activity and the enzymatic activity of acid phosphatase. These results therefore verify the important role of Cordyceps polysaccharide in triggering such immune responses. 虫草免疫价值的重新认识与实证研究

CORDYCEPS SINENSIS: A precious parasitic fungus infecting Tibet Field Mycology

Exopolysaccharide production in batch and semi-continuous fermentation of Cordyceps sinensis Biotechnology

Analysis of sterols and fatty acids in natural and cultured Cordyceps by one-step derivatization followed with gas chromatography–mass spectrometry Pharmaceutical and Biomedical Analysis
Ten free fatty acids namely lauric acid, myristic acid, pentadecanoic acid, palmitoleic acid, palmitic acid, linoleic acid, oleic acid, stearic acid, docosanoic acid and lignoceric acid and four free sterols including ergosterol, cholesterol, campesterol and β-sitosterol in natural (wild) Cordyceps sinensis, Cordyceps liangshanensis and Cordyceps gunnii, as well as cultured C. sinensis and Cordyceps militaris were first

determined using pressurized liquid extraction (PLE), trimethylsilyl (TMS) derivatization and GC–MS analysis. The conditions such as the amount of reagent, temperature and time for TMS derivatization of analytes were optimized. Under the optimum conditions, all calibration curves showed good linearity within the tested ranges. The intra- and inter-day variations for 14 investigated compounds were less than 3.4% and 5.2%, respectively. The results showed that palmitic acid, linoleic acid, oleic acid, stearic acid and ergosterol are main components in natural and cultured Cordyceps which could be discriminated by hierarchical clustering analysis based on the contents of 14 investigated compounds or the 4 fatty acids, where the contents of palmitic acid and oleic acid in natural Cordyceps are significantly higher than those in the cultured ones.

Chromatography Determination of nucleotides, nucleosides and their transformation products in Cordyceps by ion-pairing reversed-phase liquid chromatography–mass spectrometry
An ion-pairing reversed-phase liquid chromatography–mass spectrometry (IP-RP-LC–MS) was developed for the determination of nucleotides, nucleosides and their transformation products in Cordyceps. Perfluorinated carboxylic acid, namely pentadecafluorooctanoic acid (PDFOA, 0.25 mM), was used as volatile ion-paring agent and a reversed-phase column (Agilent ZORBAX SB-Aq column) was used for the separation of three nucleotides namely uridine-5′-monophosphate (UMP, 0.638–10.200 g/mL), adenosine-5′-monophosphate (AMP, 0.24–7.80 g/mL) and

guanosine-5′-monophosphate (GMP, 0.42–13.50 g/mL), seven nucleosides including adenosine (0.55–8.85 g/mL), (0.21–6.60 g/mL), guanosine cytidine (0.42–6.75 g/mL), uridine (0.33–10.50 g/mL), and inosine cordycepin

(0.48–15.30 g/mL),

thymidine

(0.20–6.30 g/mL)

(0.09–1.50 g/mL), as well as six nucleobases, adenine (0.22–6.90 g/mL), guanine (0.26–4.20 g/mL), uracil (0.38–12.15 g/mL), hypoxanthine (0.13–4.20 g/mL), cytosine (0.39–12.45 g/mL) and thymine (0.26–8.25 g/mL) with 5-chlorocytosine arabinoside as the internal standard. The overall LODs and LOQs were between 0.01–0.16 g/mL and 0.04–0.41 g/mL for the 16 analytes, respectively. The contents of 16 investigated compounds in natural and cultured Cordyceps were also determined and compared after validation of the developed IP-RP-LC-MS method. The transformations of nucleotides and nucleosides in Cordyceps were evaluated based on the quantification of the investigated compounds in three extracts, including boiling water extraction (BWE), 24 h ambient temperature water immersion (ATWE) and 56 h ATWE extracts. Two transformation pathways including UMP → uridine → uracil and GMP → guanosine → guanine were proposed in both natural Cordyceps sinensis and cultured Cordyceps militaris. The pathway of AMP → adenosine → inosine → hypoxanthine was proposed in natural C. sinensis, while AMP → adenosine → adenine in cultured C. militaris. However, the transformation of nucleotides and nucleosides was not found in commercial cultured C. sinensis.




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