Cordycepin, a metabolite of Cordyceps militaris, inhibits xenograft tumor growth of tongue squamous cell carcinoma in nude mice / 南方医科大学学报

OBJECTIVE@#To evaluate the inhibitory effect of cordycepin on oral cancer xenograft in nude mice and explore the underlying mechanisms.@*METHODS@#Sixteen BALB/c mice bearing subcutaneous human tongue squamous cell carcinoma (TSCC) TCA-8113 cell xenografts were randomized into model group and cordycepin treatment group for daily treatment with saline and cordycepin for 4 weeks. After the treatment, the tumor xenografts were dissected and weighed to assess the tumor inhibition rate. Histological changes in the heart, spleen, liver, kidney, and lung of the mice were evaluated with HE staining, and tumor cell apoptosis was examined using TUNEL staining; The expressions of Bax, Bcl-2, GRP78, CHOP, and caspase-12 in the xenografts were detected using RT-qPCR and Western blotting.@*RESULTS@#Cordycepin treatment resulted in a tumor inhibition rate of 56.09% in the nude mouse models, induced obvious changes in tumor cell morphology and significantly enhanced apoptotic death of the tumor cells without causing pathological changes in the vital organs. Cordycepin treatment also significantly reduced Bcl-2 expression (P < 0.05) and increased Bax, GRP78, CHOP, and caspase-12 expressions at both the RNA and protein levels in the tumor tissues.@*CONCLUSION@#Cordycepin treatment can induce apoptotic death of TCA-8113 cell xenografts in nude mice via the endogenous mitochondrial pathway and endoplasmic reticulum stress pathways.

Construction and optimization of cordycepin-producing Saccharomyces cerevisiae / 生物工程学报

Cordycepin is the key active component of medicinal fungus Cordyceps militaris, and it shows multiple functional activities such as anti-tumor and anti-virus. Cordycepin was conventionally produced by liquid fermentation of C. militaris, but the long production cycle and the low productivity constrained its development and application. In this study, two key genes for cordycepin biosynthesis (ScCNS1 and ScCNS2) were introduced into Saccharomyces cerevisiae S288C, producing 67.32 mg/L cordycepin at 240 h. Analysis of gene expression profiles indicated that ZWF1, PRS4, ADE4, ScCNS1 and ScCNS2 which encode enzymes involved in pentose phosphate pathway, purine metabolism and cordycepin biosynthesis pathway, were significantly up-regulated in the late phage of fermentation. Optimization of fermentation medium determined that 50 g/L initial glucose followed by feeding, supplemented with 5 mmol/L Cu²⁺ and 1.0 g/L adenine were the best condition. Fed-batch fermentation using the engineered yeast in a 5 L stirred fermenter produced 137.27 mg/L cordycepin at 144 h, with a productivity up to 0.95 mg/(L·h) reached, which was 240% higher than that of the control.

Synergistic Effect of Acoustic and Vacuum Drying to Antioxidant Attributes of Cordyceps militaris

Acoustic drying allows the utilization of lower temperatures than conventional methodology in the drying process.Vacuum drying is one of the most energy demanding processes. Water evaporation also takes place at lower temperatures under vacuumand hence the product processing temperature can be significantly lower, offering higher product quality. Cordyceps militarisis a well-known entamophagus fungus with wonderful health benefits such as adaptogenic, aphrodisiac, anti-oxidant, anti-aging, neuroprotective, nootropic, immunomodulatory, anti-cancer and hepatoprotective role by its phytochemical constituents. This study focused on the synergistic effects of acoustic and vacuum drying on antioxidant properties of Cordyceps militaris. We noticed that acoustic drying at power 800 W in frequency 40kHz combined with vacuum drying at pressure -0.8 bar were suitable for dehydration of this valuable material. From this approach, a combination of acoustic and vacuum drying created a synergistic effect consuming less energy than single drying method because it can be performed at low temperature while maintaining the product quality and wholesomeness. Moisture content is partly removed by acoustic drying and further dehydration in a vacuum dryer to reduce moisture to a stable level

Fermentation of mulberry leaves with Cordyceps militaris enhanced anti-adipogenesis activity in 3T3-L1 cells through down-regulation of PPAR-? pathway signaling

Objective: To establish an efficacious and efficient fermentation method of enhancing the anti-adipogenesis effect of mulberry (Morus alba) leaves using Cordyceps militais. Methods: Dried mulberry leaves, dried mulberry leaves with 50% raw silkworm pupa and raw silkworm pupa were fermented with Cordyceps militais for 4 weeks at 25 °C, after which the dried mulberry leaves and fermented product were extracted with 70% ethanol and subjected to high performance liquid chromatography (HPLC). The contents of cordycepin, pelargonidin, chlorogenic acid, iso-quercetin and caffeic acid were determined. We then used the 3T3-L1 cells to investigate whether extracts of fermentation enhanced anti-adipogenesis activity in vitro. Results: HPLC showed that fermentation changed the contents of cordycepin, pelargonidin, chlorogenic acid, iso-quercetin and caffeic acid. Furthermore, fermented dried mulberry leaves with 50% raw silkworm pupa had a better efficacy of anti-adipogenesis than dried mulberry leaves, fermented dried mulberry leaves and fermented silkworm pupa and inhibited triglycerides accumulation and glucose consumption. Additionally, fermented dried mulberry leaves with 50% raw silkworm pupa inhibited PPAR-? signaling. Conclusions: Fermentation with Cordyceps militaris enhanced anti-adipogenesis efficacy of mulberry leaves.

In vitro bioactive potential and chemical analysis of the n-hexane extract of the medicinal mushroom, Cordyceps militaris

@#Aims: Cordyceps militaris is a medicinal mushroom from Ascomycota. The aims of this study were to explore and identify the chemical compounds extracted in the non-polar fraction of the mushroom and to examine the biological potential of this extract. Methodology and results: The n-hexane extract metabolites were identified using gas chromatography-mass spectrometry (GC-MS) and results revealed the presence of 37 compounds delivered from different chemical classes and were mainly comprised of fatty acids and their esters (72%), carboxylic acids and their esters (10.39%), and a sulphur compound (7.1%). The n-hexane extract recorded a promising antioxidant effect (80.9±1.5%) at 80 mg/mL total extract; potent cholesterol reduction activity (100%) was obtained after 96 h incubation by the total metabolites (4%). The cytotoxicity of the compounds revealed 50% cytotoxicity concentration (CC50) ˃ 1 mg/mL and anti-rotavirus SA-11 effect where inhibition of virus attachment and penetration into infected cells was recorded at 50% effective concentration (IC50) of 300±0.2 µg/mL. Conclusion, significance and impact of study: This study confirmed the impact of the fatty acids produced by C. militaris as bioactive metabolites.

Advances in biosynthesis of cordycepin from Cordyceps militaris / 生物工程学报

Cordycepin as the main active ingredient of Cordyceps militaris, a traditional medicinal fungus in China, has many physiological functions such as anti-cancer, anti-tumor and anti-virus activity. The most potential route for effective cordycepin production has been considered as liquid fermentation of C. militaris though with low productivity at present. Thus, it is urgent to apply both process engineering strategy and metabolic engineering strategy to enhance the productivity of cordycepin. In this review, the effects of medium components (i.e. the carbon/nitrogen source, precursor substances and metal ions) and operation factors (i.e. pH, dissolved oxygen and light) on cordycepin biosynthesis in liquid fermentation system are summarized. Besides, separation of cordycepin, the gene cluster involved and predicted biosynthesis pathways of cordycepin are also discussed, providing possible solutions of finally realizing efficient production of cordycepin.

Fermentation of mulberry leaves with Cordyceps militaris enhanced anti-adipogenesis activity in 3T3-L1 cells through down-regulation of PPAR-? pathway signaling

Objective: To establish an efficacious and efficient fermentation method of enhancing the anti-adipogenesis effect of mulberry (Morus alba) leaves using Cordyceps militais. Methods: Dried mulberry leaves, dried mulberry leaves with 50% raw silkworm pupa and raw silkworm pupa were fermented with Cordyceps militais for 4 weeks at 25 °C, after which the dried mulberry leaves and fermented product were extracted with 70% ethanol and subjected to high performance liquid chromatography (HPLC). The contents of cordycepin, pelargonidin, chlorogenic acid, iso-quercetin and caffeic acid were determined. We then used the 3T3-L1 cells to investigate whether extracts of fermentation enhanced anti-adipogenesis activity in vitro. Results: HPLC showed that fermentation changed the contents of cordycepin, pelargonidin, chlorogenic acid, iso-quercetin and caffeic acid. Furthermore, fermented dried mulberry leaves with 50% raw silkworm pupa had a better efficacy of anti-adipogenesis than dried mulberry leaves, fermented dried mulberry leaves and fermented silkworm pupa and inhibited triglycerides accumulation and glucose consumption. Additionally, fermented dried mulberry leaves with 50% raw silkworm pupa inhibited PPAR-? signaling. Conclusions: Fermentation with Cordyceps militaris enhanced anti-adipogenesis efficacy of mulberry leaves.

Comparison of effects of different levels of cordyceps militaris power on non-specific immune function of immunodeficient mice / 中国药理学通报

Aim: To compare the immunomodulatory effects of Super V Cordyceps Militaris (Super V CM) and I star Cordyceps Militaris (I star CM) powder on the immune suppression induced by dexamethasone in mice. Methods: Fifty male BALB/c mice were randomly divided into five groups; control and model group, Super V CM (1 g · kg-1) group, I star CM (1 g · kg-1) group and Cordyceps capsule (1 g · kg-1) group. Apart from control group, mice of the remaining groups were injected with dexamethasone 20 mg · kg-1 on 22nd and 23rd day of the experiment. Dissection was performed 24h after the last administration, then blood routine and viscera index, the function of NK cells and abdominal macrophages and the cytokine secretion levels were analyzed. Results: Dexamethasone could significantly decrease the body mass, organ index and cellular function of immune cells in mice. The Super V CM and I star CM had significant improvement in all aspects of the immunodeficiency induced by dexamethasone. Super V CM was significantly superior to I star CM in terms of improving the spleen index, enhancing the killing ability of NK cells, and improving the secretion level of TNF-a by macrophages. Conclusion: The effect of Super V CM is better than that of I star CM in enhancing non-specific immunity.

Protective effect of Cordyceps militaris against hydrogen peroxide-induced oxidative stress in vitro

BACKGROUND/OBJECTIVES: Excessive production of reactive oxygen species (ROS) such as hydroxyl (·OH), nitric oxide (NO), and hydrogen peroxide (H2O2) is reported to induce oxidative stress. ROS generated by oxidative stress can potentially damage glial cells in the nervous system. Cordyceps militaris (CM), a kind of natural herb widely found in East Asia. In this study, we investigated the free radical scavenging activity of the CM extract and its neuroprotective effects in H2O2-induced C6 glial cells. MATERIALS/METHODS: The ethanol extract of CM (100–1,000 µg/mL) was used to measure DPPH, ·OH, and NO radical scavenging activities. In addition, hydrogen peroxide (H2O2)-induced C6 glial cells were treated with CM at 0.5–2.5 µg/mL for measurement of cell viability, ROS production, and protein expression resulting from oxidative stress. RESULTS: The CM extract showed high scavenging activities against DPPH, ·OH, and NO radicals at concentration of 1,000 µg/mL. Treatment of CM with H2O2-induced oxidative stress in C6 glial cells significantly increased cell viability, and decreased ROS production. Cyclooxygenase-2 and inducible nitric oxide synthase protein expression was down-regulated in CM-treated groups. In addition, the protein expression level of phospho-p38 mitogen-activated protein kinase (p-p38 MAPK), phospho-c-Jun N-terminal kinase (p-JNK), and phospho-extracellular regulated protein kinases (p-ERK) in H2O2-induced C6 glial cells was down-regulated upon CM administration. CONCLUSION: CM exhibited radical scavenging activity and protective effect against H2O2 as indicated by the increased cell viability, decreased ROS production, down-regulation of inflammation-related proteins as well as p-p38, p-JNK, and p-ERK protein levels. Therefore, we suggest that CM could play the protective role from oxidative stress in glial cells.

Production of antioxidative exopolysaccharides of Cordyceps militaris with Vernonia amygdalina leaves in substrate / 生物工程学报

Cordyceps militaris exopolysaccharides (EPS) have many pharmacological activities such as boosting immunity and antifatigue. To obtain EPS efficiently, we added moderate Vernonia amygdalina leaf powder as inducer to the fermentation medium to promote the production of Cordyceps militaris EPS and studied the infrared absorption spectrum and antioxidant activities of the EPS after optimization. The optimum liquid fermentation conditions were as follows: addition of Vernonia amygdalina leaf powder of 8 g/L, fermentation duration of 9 d, initial pH of 6.5, inoculation quantity of 5.0 mL. Under such a condition, the yield of Cordyceps militaris EPS reached (5.24±0.28) mg/mL, increased by 205.20% compared to the control group without adding Vernonia amygdalina leaf powder. Results of infrared analysis and antioxidant activity showed that the Vernonia amygdalina leaves had little effect on the structure and activities of Cordyceps militaris EPS. The results of this research suggest that Vernonia amygdalina leaf can enhance the production of Cordyceps militaris EPS effectively, and provides a novel method for efficient production of EPS in Cordyceps militaris.

Effects of Cordyceps militaris supplementation on the immune response and upper respiratory infection in healthy adults: a randomized, double-blind, placebo-controlled study / 한국영양학회지

PURPOSE: Upper respiratory tract infections are major causes of the common cold throughout the world. Cordyceps militaris (C. militaris) is a well-known functional food for its anti-fatigue and immunomodulating activities. On the other hand, there are no reports on the protective effect against upper respiratory tract infections (URI). This study was a 12 week randomized, double-blind, and placebo-controlled trial in healthy volunteers. METHODS: A total of 100 subjects 20 ~ 70 years of age with a history of at least two colds in the year were enrolled in the study. The participants were required to record any adverse events and rate any cold-related incidents in a diary during the investigation period. The efficacy end point was the symptoms and incidence of URI, and changes in cytokines, IgA and natural killer (NK) cell activity. RESULTS: The Cordyceps militaris group over 12 weeks showed no significant impact on the incidence and symptomatology of URI compared to the placebo group. On the other hand, the experimental group showed significantly higher NK cell activity (p = 0.047) and IgA level (p = 0.035) compared to the placebo group. The NK-cell activity and IgA level were increased significantly by Cordyceps militaris over 12 weeks. CONCLUSION: The results suggest the possible beneficial immunomodulating effects, but the protective effects on URI could not be demonstrated under these conditions. Additional research will be needed to determine the efficacy and mechanisms of Cordyceps militaris function.

Optimization of liquid fermentation conditions for biotransformation zein by Cordyceps militaris 202 and characterization of physicochemical and functional properties of fermentative hydrolysates

Abstract Cordyceps militaris 202 is a potential fungus for biotransformation zein, due to its various proteases, high tolerance and viability in nature. In this article, single factor experiment and response surface methodology were applied to optimize the liquid fermentation conditions and improve the ability of biotransformation zein. The optimized fermentation conditions were as follows: inoculum concentration of 19%, volume of liquor of 130 mL/500 mL and pH of 4.7. Under this condition, the degree of hydrolysis (DH) was 27.31%. The zein hydrolysates from fungi fermentation maintained a high thermal stability. Compared to the original zein, the zein hydrolysates were found to have high solubility, which most likely results in improved foaming and emulsifying properties. Overall, this research demonstrates that hydrolysis of zein by C. militaris 202 is a potential method for improving the functional properties of zein, and the zein hydrolysates can be used as functional ingredients with an increased antioxidant effect in both food and non-food applications.

Quantitative analysis of five nucleosides in Cordyceps by HPLC and quality control of Cordyceps militaris / 中草药

Objective To establish an HPLC method for simultaneous determination of uridine, vernine, adenosine, cordycepin, and N6-(2-hydroxyethyl)-adenosine in Cordyceps (Cordyceps militaris, Cordyceps Fungus Powder CS-4, Hirsutella sinensis, and C. sinensis), and determine the characteristic components of C. militaris to provide the scientific basis for quality control of C. militaris and its extract. Methods HPLC was performed on Inertsil ODS-3 column (250 mm × 4.6 mm, 5 μm) with mobile phase A (acetonitrile) and B (water) for gradient elution. The flow rate was 0.8 mL/min, detection wavelength was 260 nm, and column temperature was 30 ℃. Results All mentioned five nucleosides can be detected from C. militaris. However, cordycepin and N6-(2-hydroxyethyl)-adenosine were undetected in the other three Cordyceps species. The sample preparation method of C. militaris has a great effect on the content of nucleosides. The content of uridine, vernine and adenosine was the highest in samples prepared by ultrasonic extraction 180 min. Cordycepin was stable form six sample preparation methods, and N6-(2-hydroxyethyl)-adenosine was unbearable to heat and acid. The contents of cordycepin and N6-(2-hydroxyethyl)-adenosine were the same in four preparation methods. Conclusion This experiment provides a basis for the quality analysis of C. militaris and its extracts. Cordycepin and N6-(2-hydroxyethyl)-adenosine can be used as markers for the quality control of C. militaris and its extracts.

Scratching Stimuli of Mycelia Influence Fruiting Body Production and ROS-Scavenging Gene Expression of Cordyceps militaris

The entomopathogenic fungus Cordyceps militaris is a valuable medicinal ascomycete, which degenerates frequently during subsequent culture. To avoid economic losses during industrialized production, scratching stimuli of mycelia was introduced to improve the fruiting body production. The present results indicated that higher yields and biological efficiency were obtained from two degenerate strains (YN1-14 and YN2-7) but not from g38 (an insertional mutant in Rhf1 gene with higher yields and shorter growth periods). Furthermore, the growth periods of the fruiting bodies were at least 5 days earlier when the mycelia were scratched before stromata differentiation. Three ROS-scavenging genes including Cu/Zn superoxide dismutase (CmSod1), Glutathione peroxidase (CmGpx), and Catalase A (CmCat A) were isolated and their expression profiles against scratching were determined in degenerate strain YN1-14 and mutant strain g38. At day 5 after scratching, the expression level of CmGpx significantly decreased for strain g38, but that of CmSod1 significantly increased for YN1-14. These results indicated that scratching is an effective way to promote fruiting body production of degenerate strain, which may be related at least with Rhf1 and active oxygen scavenging genes.

Echinacoside and Cistanche tubulosa(Schenk)R.wight have male reproductiveprotection effect by targeting hypothalamic and rogen receptor / 中国药理学与毒理学杂志

OBJECTIVE Cistanche deserticola Y. C. Ma is an official plant that grows in arid or semi-arid areas.Our early work demonstrated that Cistanche extracts protect against sperm damage in mice under bisphenol A induced reproductive damage. Echinacoside (ECH)is one of the major active components. The present study investigated the mechanisms behind the possible protective effects of ECH against oligoasthenospermia in rat and identified the interaction between ECH and AR. METH-ODS The distribution of ECH was assayed by HPLC,the quantity and quality of sperm was evaluated and hormone levels were determined by radioimmunosorbent assay. The levels of androgen receptor (AR)and key steroidogenic-related genes were reduced as determined by Western blotting and qPCR analysis.The interaction between ECH and AR were evaluated by fluorescence localization assay,indi-rect ELISA and molecular docking. RESULTS ECH significantly increased the quantity of sperm and secretions of luteinizing hormone and testosterone.ECH was distributed to the hypothalamus but not in the testes.ECH combined with hypothalamic AR in the pocket of Met-894 and Val-713 to inhibit transfer of AR from the cytoplasm to nuclei in the hypothalamus.While negative feedback of sex hormone regula-tion was inhibited, positive feedback was stimulated to increase the secretion of luteinizing hormone and testosterone subsequently enhancing the quantity of sperm. C. militaris significantly alleviated the BPA-induced reproductive damage by increasing testicular superoxide dismutase (SOD), glutathione peroxidase (GSH-PX) and glutathione (GSH); as well as by reducing serum malondialdehyde (MDA). C. militaris not only obviously enhanced the levels of serum LH and T, but it also improved the sperm count and motility compared to the BPA-treated group.CONCLUSION C.militaris and ECH protect the BPA induced reproductive damage.ECH blocks AR activity in the hypothalamus to increase the quantity of sperm and protect against oligoasthenospermia in rats.

Simultaneous Determination of 5 Nucleosides from Different Parts of Cultured Cordyceps militaris by HPLC / 中国药房

OBJECTIVE:To establish a method for simultaneous determination of 5 nucleoside from different parts of cultured Cordyceps militaris. METHODS:HPLC method was adopted. The determination was performed on InertSustain C18 column with mobile phase consisting of methanol-0.02 mol/L monobasic potassium phosphate solution (gradient elution) at the flow rate of 0.6 mL/min. The detection wavelength was set at 254 nm,the column temperature was 30 ℃. RESULTS:The linear ranges of uridine, inosine,guanosine,adenosine and cordycepin were 0.568-3.408 μg(r=0.9999),0.284-1.704 μg(r=0.9999),0.264-1.584 μg(r=0.9999),0.232-1.392 μg(r=0.9999)and 1.672-10.032 μg(r=0.9998),respectively. RSDs of precision,stability and repeatabili-ty tests were all lower than 3.0%. The recoveries were 98.2%-103.9%(RSD=1.97%,n=9),96.2%-101.6%(RSD=1.76%,n=9),96.7%-102.0%(RSD=1.94%,n=9),95.1%-99.4%(RSD=1.43%,n=9) and 95.6%-101.3%(RSD=1.82%,n=9). CON-CLUSIONS:The method is simple,precise,stable and repeatable,and can be used for simultaneous determination of 5 nucleoside from cultured C. militaris,the content of nucleosides are different in different parts of C. militaris.

Simultaneous Determination of Contents of 4 Heavy Metal Elements in Cultured Cordyceps militaris by ICP-MS / 中国药房

OBJECTIVE:To establish the method for the simultaneous determination of contents of 4 heavy metal elements in cultured Cordyceps militaris. METHODS:The samples were digested by nitric acid heating method and then determined by ICP-MS including radio-frequency power of 1.15 kW,sampling depth of 65 nm,auxiliary gas flow rate of 1.0 L/min,cooling gas flow of 13.0 L/min,temperature of 25 ℃,automatic plasma. RESULTS:The linear ranges of arsenic(As),cadmium(Cd),mer-cury (Hg) and lead (Pb) were 0-20 μg/mL(r=0.9970),0-10 μg/mL(r=0.9995),0-5 μg/mL(r=0.9955),0-20 μg/mL(r=0.9960),respectively. Detectio limit were 0.128,0.003,0.002,0.004 mg. RSDs of precision,stability and reproducibility tests were all lower than 6.5%. Recoveries of them were 90.4%-100.6%(RSD=3.45%,n=9),94.3%-101.3%(RSD=2.93%,n=9), 90.0%-102.3%(RSD=4.03%,n=9),92.3%-103.0%(RSD=3.53%,n=9),respectively. CONCLUSIONS:The method is sim-ple,precise,stable,reproducible,and can be used for simultaneous determination of contents of 4 heavy metal elements in cul-tured C. militaris. The contents of As,Cd,Hg and Pb in 10 batches of sample are all in line with related national standards.

Enhancement effect of Superfine Cordyceps militarisPowder on immune functions of mice / 吉林大学学报(医学版)

Objective:To investigate the effect of Superfine Cordyceps militarisv Powder on the immune functions of the mice,and to provide basis for improving the utilizable ratio of Cordyceps militaris.Methods:A total of 40 mice were randomly divided into negative control group,low dose (0.166 5 g·kg-1),middle dose (0.333 3 g·kg-1),and high dose (0.999 9 g·kg-1) of Superfine Cordyceps militaris Powder groups,and there were 10 mice in each group.The mice in different doses of Superfine Cordyceps militaris Powder groups were administered with the Superfine Cordyceps militaris Powder for 30 d by intragastric infusion respectively,whereas,the mice in negative control group were administered with the same volume of water for 30 d by intragastric infusion.The body weights,the spleen indexes and thymus indexes of the mice in various groups were measured;the lymphocyte transformation abilities of the mice in various groups were observed by lymphocyte transformation experiment;the levels of delayed-type hypersensitivity reaction of the mice were examined with toe incrassation;the number of antibody forming cells,phagocytic rate and phagocytic index of chicken erythrocytes phagocytized by peritoneal macrophages of the mice were detected.Results:Compared with negative control group,the body weights,the spleen indexes a nd thymus indexes of the mice in different doses of Superfine Cordyceps militaris Powder groups had no significant differences (P>0.05).The lymphocyte transformation abilities of the mice in middle and high doses of Superfine Cordyceps militarisPowder groups were higher than that in negative control group(P<0.05).The levels of delayed-type hypersensitivity reaction of the mice in middle and high doses of Superfine Cordyceps militaris Powder groups were higher than those in low dose of Superfine Cordyceps militaris powder group and negative control group(P<0.05).The numbers of antibody forming cells of the mice in low,middle and high doses of Superfine Cordyceps militaris Powder groups were higher than that in negative control group(P<0.05).The phagocytic rates and phagocytic indexes of chicken erythrocytes phagocytized by peritoneal macrophage of the mice in low,middle and high doses of Superfine Cordyceps militaris Powder groups were higher than that in negative control group(P<0.05).Conclusion:Superfine Cordyceps militaris Powder can strengthen the immune functions of the mice,and the recommended doses are 0.333 3 and 0.999 9 g·kg-1.

Comparative Study on the Contents of Fatty Acids in Isaria cicadae, Cordyceps militaris and Cordyceps sinensis / 中国药房

OBJECTIVE:To establish a method for simultaneous determination of fatty acids in Isaria cicadae,Cordyceps militaris and C.sinensis,and to compare the difference of the contents of fatty acids among above medicinal herbs.METHODS:GC-MS method was adopted.Chromatographic condition:the determination was performed on TG-5MS gas phase capillary column with carrier gas of nitrogen at the flow rate of 1.2 mL/min.The inlet temperature was 290 ℃ by splitlesssampling;valve opening time was 1 min,and volume of sample was 1 μL.Mass spectrometry condition:the ion source is an electrospray ionization source.The temperatures of ion source and transmission line were 280 ℃,the initial temperature of the chromatographic column was 80 ℃ (gradient elution),ionization voltage was 70 eV.Solvent delay time was 5 min,and scan mass range was m/z 30-400.RESULTS:The linear ranges of tetradecanoic acid,pentadecanoic acid,palmitic acid,palmitoleic acid,heptadecanoic acid,heptadecenoic acid,docosahexaenoic acid,methyl oleate,linoleic acid,arachidonic acid,eicosenoic acid,diolefinic acid,eicosatrienoic acid,heneicosanoic acid,behenic acid,tricosanoic acid,lignoceric acid were 1.400-44.520 μg/mL(r=0.999 8),2.091-93.721 μg/mL(r=0.999 7),3.146-85.856 μg/mL(r=0.998 2),1.664-61.444 g/rnL(r=0.998 7),1.773-64.983 g/mL(r=0.999 5),1.781-68.421 μg/ mL (r=0.999 7),1.706-55.606 μg/mL (r=0.999 8),1.439-47.989 μg/mL (r=0.999 6),1.738-66.908 μg/mL (r=0.999 6),2.086-94.206 μg/mL(r=0.999 5),1.356-44.966 μg/mL(r=0.999 4),1.444-56.814 μg/mL(r=0.999 7),1.375-52.335 μg/mL(r =0.999 8),1.512-60.312 μg/mL(r=0.999 5),1.450-59.760 μg/mL(r=0.999 7),1.427-58.757 μg/mL(r=0.999 1),1.269-58.109 μg/ mL(r=0.999 3),respectively.The limit of quantitation was no more than 1 764.71 μg/mL,and the limit of detection was no more than 529.42 μg/mL.RSDs of precision,stability and reproducibility tests were all lower than 2.0％.The recoveries were 84.87％-108.93％ (RSD ranged 0.19％-2.23％,n=6).There were 17 fatty acids in C.militaris,16 fatty acids in Ⅰ.cicadae and 16 fatty acids in C.sinensis.The contents of unsaturated fatty acids in above medicinal herbs were higher than that of saturated fatty acids.The content of fatty acids in artificial cultivated Ⅰ.cicadae was mostly higher than other medicinal herbs.COCLUSIONS:The method is simple,accurate,stable and reproducible.It can be used for simultaneous determination of fatty acids in I.cicadae,C.militaris and C.sinensis.Above 3 kinds of medicinal materials.From the perspective of fatty acid content,the quality of artificial cultivated I.cicadae is best.

HPLC fingerprint of Cordyceps sinensis and its confused species and identification of common composition / 中草药

Objective: To establish an HPLC fingerprint method for water-soluble components in Cordyceps sinensis which could provide the basis for authenticity identification and quality evaluation of C. sinensis. Methods: HPLC method was adopted to construct the fingerprint of water soluble components in C. sinensis and its main confused species (C. liangshanensis, C. gunnii and artificial C. militaris). Total 16 common peaks were marked, and 12 common peaks were confirmed by the method with tandem high performance liquid chromatography (HPLC) and quadrupole time-of-flight mass spectrometry (Q-TOF MS). Results: The common mode of HPLC fingerprint for C. sinensis was developed. The method could distinguish between C. sinensis and its confused species, and 12 common peaks were identified, including cytosine, uracil, cytidine, hypoxanthine + guanine, uridine, thymine, adenine, inosine, guanosine, thymidine, adenosine, and cordycepin respectively. Conclusion: This method is simple and easy, providing scientific basis for identification of C. sinensis.