ABSTRACT
GI (GIGANTEA) is one of the output key genes for circadian clock in the plant. The JrGI gene was cloned and its expression in different tissues was analyzed to facilitate the functional research of JrGI. RT-PCR (reverse transcription-polymerase chain reaction) was used to clone JrGI gene in present study. This gene was then analyzed by bioinformatics, subcellular localization and gene expression. The coding sequence (CDS) full length of JrGI gene was 3 516 bp, encoding 1 171 amino acids with a molecular mass of 128.60 kDa and a theoretical isoelectric point of 6.13. It was a hydrophilic protein. Phylogenetic analysis showed that JrGI of 'Xinxin 2' was highly homologous to GI of Populus euphratica. The result of subcellular localization showed that JrGI protein was located in nucleus. The JrGI, JrCO and JrFT genes in female flower buds undifferentiated and early differentiated of 'Xinxin 2' were analyzed by RT-qPCR (real-time quantitative PCR). The results showed that the expression of JrGI, JrCO and JrFT genes were the highest on morphological differentiation, implying the temporal and special regulation of JrGI in the differential process of female flower buds of'Xinxin 2'. In addition, RT-qPCR analysis showed that JrGI gene was expressed in all tissues examined, whereas the expression level in leaves was the highest. It is suggested that JrGI gene plays a key role in the development of walnut leaves.
Subject(s)
Juglans/genetics , Phylogeny , Plant Leaves , Cloning, Molecular , Gene Expression Regulation, Plant , Plant Proteins/metabolismABSTRACT
A resistência antimicrobiana atingiu proporções alarmantes em todo o mundo, sendo que na Europa mortes causadas por micro-organismos multirresistentes superam os índices de mortalidade da AIDS, tuberculose e a gripe. Assim a fitoterapia desponta no combate a esta problemática, com as diversas atividades biológicas de plantas e seus derivados. Portanto os objetivos do presente estudo foram avaliar a ação antimicrobiana, anti-inflamatória, citotoxicidade, genotoxicidade e constituição fitoquímica dos extratos glicólicos de P. paniculata e J. regia. A ação sobre bactérias anaeróbias (Porphyromonas gingivalis, P. endodontalis, Parvimonas micra e Fusobacterium nucleatum) foi realizada por meio dos testes de microdiluição em caldo (Protocolo M11-A8 - CLSI) e sobre biofilmes monotípicos. Já a ação sobre aeróbios foi realizada sobre 3 cepas de Klebsiella pneumoniae multirresistente, com testes sobre culturas planctônicas (Protocolo M7-A9) e biofilmes; Foi realizada a verificação da atividade antimicrobiana sobre biofilmes heterotípicos de Candida albicans associada a Staphylococcus aureus, Enterococcus faecalis, Streptococcus mutans ou Pseudomonas aeruginosa. A citotoxicidade e a genotoxicdade dos extratos foram avaliadas sobre macrófagos de camundongos (RAW 264.7) e queratinócitos humanos (HaCat) pelos testes de MTT e micronúcleos, respectivamente. O potencial antiinflamatório foi verificado dosando os níveis de TNF-âº, IL-10 e IL-1ß pelo teste de ELISA. Os dados obtiveram distribuição normal sendo a análise estatística realizada por ANOVA e Tukey (p<0,05%). Os extratos de P. paniculata e J. regia promoveram CMM de 50 mg/mL para as anaeróbias. Os biofilmes de P. gingivalis e P. micra foram eliminados com 100 e 200 mg/mL dos extratos (5 min) e com as concentrações de 50 e 100 mg/mL por 24 h; F. nucleatum e P. endodontalis obtiveram reduções variando de 80 a 90%. Os biofilmes heterotípicos de C. albicans e S. mutans obtiveram reduções de até 80% após contato por 5 min. com J. regia e 71% para P. paniculata. Os biofilmes multirresistentes de K. pneumoniae obtiveram reduções na atividade metabólica de até 67,9%. P. paniculata promoveu viabilidade celular variando de 61,1% a 133,8% sobre queratinócitos humanos após 24 h de contato com as concentrações de 12,5 a 0,39 mg/mL, enquanto J. regia obteve 43,9 a 128,4% de viabilidade. Os macrófagos de camundongo obtiveram viabilidade de 18,1 a 101,9% com P. paniculata e 35,4 a 60,6% para J. regia. P. paniculata promoveu a redução nos níveis da citocina pró-inflamatória IL-1ß e aumento nos níveis da citocina antiinflamatória IL-10. Já J. regia promoveu a redução da citocina pró-inflamatória TNF-âº. Ambos os extratos não promoveram genotoxicidade frente as linhagens celulares. A análise fitoquímica evidenciou a presença de benzofenonas e ácido cafeoilquínico nos extratos de P. paniculata e J. regia, respectivamente. Em conclusão, os extratos de P. paniculata e J. regia demonstraram ação antimicrobiana sobre bactérias aeróbias e anaeróbias e multirresistentes com destaque a eliminação dos biofilmes de P. gingivalis, P. endodontalis, P. micra e K. pneumoniae (multirresistentes). Os extratos demonstraram a ausência de toxicidade e genotoxicidade conforme tempo de aplicação e concentração utilizada, além de possuírem potencial anti-inflamatório(AU)
Antimicrobial resistance has reached alarming proportions worldwide, with deaths in Europe caused by multi-resistant microorganisms exceeding the mortality rates from AIDS, tuberculosis and influenza. Thus phytotherapy emerges in the fight against this problem, with the various biological activities of plants and their derivatives. Therefore, the objectives of the present study were to evaluate the antimicrobial, antiinflammatory, cytotoxicity, genotoxicity and phytochemical constitution of the glycolic extracts of P. paniculata and J. regia. The action on anaerobic bacteria (Porphyromonas gingivalis, P. endodontalis, Parvimonas micra and Fusobacterium nucleatum) was carried out by means of broth microdilution tests (Protocol M11-A8 - CLSI) and on monotypic biofilms. The action on aerobes was performed on 3 strains of multi-resistant Klebsiella pneumoniae, with tests on planktonic cultures (Protocol M7-A9) and biofilms; The verification of antimicrobial activity on heterotypic biofilms of Candida albicans associated with Staphylococcus aureus, Enterococcus faecalis, Streptococcus mutans or Pseudomonas aeruginosa was also performed. The cytotoxicity and genotoxicity of the extracts were evaluated on mouse macrophages (RAW 264.7) and human keratinocytes (HaCat) by MTT and micronucleus tests, respectively. The anti-inflammatory potential was assessed by the ELISA test, TNF-âº, IL-10 and IL-1ß levels were measured. The data obtained a normal distribution and the statistical analysis was performed by ANOVA and Tukey (p <0.05%). The extracts of P. paniculata and J. regia promoted CMM of 50 mg / mL for anaerobes. The biofilms of P. gingivalis and P. micra were eradicated with 100 and 200 mg / mL of the extracts (5 min) and with the concentrations of 50 and 100 mg / mL (24 hours); F. nucleatum and P. endodontalis obtained reductions ranging from 80 to 90%. The heterotypic biofilms of C. albicans and S. mutans obtained reductions of up to 80% after contact for 5 minutes with J. regia and 71% for P. paniculata. The multidrug-resistant strains of K. pneumoniae obtained reductions in metabolic activity of up to 67.9%. The P. paniculata extract promoted cell viability ranging from 61.1% to 133.8% on human keratinocytes after 24 h of contact with concentrations of 12.5 to 0.39 mg / mL, while J. regia obtained 43, 9 to 128.4% viability. Mouse macrophages obtained viability from 18.1 to 101.9% with P. paniculata and 35.4 to 60.6% for J. regia. P. paniculata promoted a reduction in the levels of the pro-inflammatory cytokine IL-1ß and an increase in the levels of the anti-inflammatory cytokine IL-10. J. regia promoted the reduction of the pro-inflammatory cytokine TNF-âº. Both extracts did not promote genotoxicity against cell lines. Phytochemical analysis showed the presence of benzophenones and caffeoylquinic acid in the extracts of P. paniculata and J. regia, respectively. In conclusion, the extracts of P. paniculata and J. regia demonstrated antimicrobial action on aerobic and anaerobic and multiresistant bacteria, with emphasis on the elimination of the biofilms of P. gingivalis, P. endodontalis and P. micra, as well as the reductions of the biofilms of K. pneumoniae multidrug-resistant. The extracts demonstrated the absence of toxicity and genotoxicity according to the time of application and concentration used, in addition to having anti-inflammatory potential(AU)
Subject(s)
Anti-Bacterial Agents/immunology , Drug Resistance, Microbial/drug effects , Juglans/adverse effects , Phytotherapy/methodsABSTRACT
BACKGROUND: Juglone is a naphthoquinone currently obtained by chemical synthesis with biological activities including antitumor activity. Additionally, juglone is present in the green husk of walnut, which suggests evaluating the effect of GH extracts on carcinogenic cell lines. RESULTS: Walnut green husk ethanolic extract was obtained as 169.1 mg juglone/100 g Green Husk and antioxidant activity (ORAC) of 44,920 µmol Trolox Equivalent/100 g DW Green Husk. At 1 µM juglone in HL-60 cell culture, green husk extract showed an antiproliferative effect, but pure juglone did not; under these conditions, normal fibroblast cells were not affected. A dose-dependent effect on mitochondrial membrane potential loss was observed. Apoptosis of HL-60 was detected at 10 µM juglone. Despite high ORAC values, neither purified juglone nor the extract showed protective effects on HL-60 cells under oxidative conditions. CONCLUSIONS: Green husk extract generates an antiproliferative effect in HL-60 cells, which is related to an induction of the early stages of apoptosis and a loss of mitochondrial membrane potential. The normal cells were not affected when juglone is present at concentrations of 1 µM, while at higher concentrations, there is loss of viability of both cancerous and healthy cells.
Subject(s)
Apoptosis , HL-60 Cells/metabolism , Juglans/chemistry , Polyphenols/metabolism , Antioxidants/metabolism , Cell Survival , Chromatography, High Pressure Liquid , Cell Culture Techniques , Membrane Potential, MitochondrialABSTRACT
Objective: To investigate the diarylheptanoids from the flower of Juglans regia. Methods: The compounds were isolated and purified by various column chromatographies, and their structures were identified by physiochemical properties and spectroscopic data. And MTT assays were performed to detect the in vitro cytotoxicity of compounds. Results: A total of 11 diarylheptanoids were isolated from the EtOH extract of the flower of J. regia, which were identified as tsaokoarylone (1), rhoiptelol C (2), (3S,7S)-3,7-dihydroxy-1-(4-hydroxyphenyl)-7-(3-methoxy-4-hydroxyphenyl)-heptane (3), 3-hydroxy-1-(3-methoxy-4-hydroxyphenyl)-7- (4-hydroxyphenyl) heptane (4), 1-(4-hydroxyphenyl)-7-(3-methoxy-4-hydroxyphenyl)-4-heptene-3-one (5), juglanin A (6), 2-oxatrycyclo[13.2.2.13,7]eicosa-3,5,7-(20),15,17,18-hexane-10,16-diol (7), 2-oxatrycyclo[13.2.2.13,7]eicosa-3,5,7-(20),15,17,18-hexane- 10-one (8), pterocarine (9), (8R)-32-methoxy-2-oxa-1(1,3),3(1,4)-dibenzenacyclodecaphane-16,8-diol (10), and juglanin B (11). Conclusion: Compounds 1-2, 4-5, and 7-8 are isolated from this genus for the first time. Compounds 1-11 are isolated from this plant for the first time, and compound 8 inhibited the proliferation of HCT-116, HepG2, BGC-823, NCI-H1650, and A2780 cancer cell lines in vitro, with IC50 values of 3.56, 2.26, 1.39, 2.62, and 1.18 μmol/L, respectively.
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JugLans regia is the main economic crop in China and an important medicinal plant, which have antibacterial, anti-oxidant, anti-tumor and hypoglycemic effects. It mainly contains a variety of active small molecule compounds such as polysaccharides, flavonoids, phenolic acids, esters, saponins and quinones.It can be used not only for the development of pharmaceuticals or functional foods, but also for its market demand in the food industry with good application base. It is a resource material with great development potential in walnuts. In this paper, the chemical composition and pharmacological effects of walnut were systematically combed, and on this basis, the industrial application and development path were analyzed to provide a basis for the development and utilization of walnut resources in China.
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OBJECTIVE:To optimize the extraction technology of polyphenols from Juglans regia branch and evaluate its anti-oxidant activity in vitro. METHODS:Using extraction amount of polyphenols from J. regia branch as response value,solvent-solid ratio,extraction temperature and ethanol volume fraction as response factors,based on single factor test,response surface method was used to optimize the extraction technology of polyphenols from J. regia branch. Using vitamin C as positive control,scaveng-ing on hydroxyl radicals and 1,1-diphenyl-2-picrylhydrazyl(DPPH)radicals and total reducing activities of polyphenols from J. re-gia branch were investigated. And its antioxidant activity in vitro was evaluated. RESULTS:Optimized extraction conditions for polyphenols from J. regia branch was as follow as solid-liquid ratio of 1:25(g/mL),extraction temperature of 50 ℃,ethanol vol-ume fraction of 70%. The extraction amount of polyphenols from J. regia branch was 9.30 mg/g(RSD=0.57%,n=3)in the veri-fication test. Clearance rate on hydroxyl radicals and DPPH radicals and total reducing activity of polyphenols from J. regia branch were respectively 50.24%,95.42%,1.118 when it was under the mass concentration of 12.0,3.0,3.0μg/mL;and the related data of vitamin C was 93.71%,46.17%,0.628 under the same mass concentration(P<0.05). CONCLUSIONS:Extraction technology of polyphenols from J. regia branch optimized by response surface method is stable and feasible;polyphenols from J. regia branch shows certain antioxidant activity in vitro.
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Objective: To study the chemical constituents in green walnut husks of Juglans regia. Methods: The chemical constituents were separated and purified by silica gel column chromatography and HPLC. Their structures were determined on the basis of spectroscopic analyses. Results: Fifteen compounds were isolated and the structures were identified as epi-dihydrophaseic acid (1), 4-butoxy-5, 8-dihydroxy-3, 4-dihydronaphthalen-1-one (2), 4-ethoxy-5, 8-dihydroxy-3, 4-dihydronaphthalen-1-one (3), myricatomento-genin (4), nodulisporone (5), 5, 8-dihydroxy-4S-methoxy-β-tethalone (6), 5-hydroxy-4-methoxy-α-naphthalen-1-one (7), isosclerone (8), 4, 5, 8-trihydroxy-1, 2, 3, 4-tetrahydronaphthalene-1-one (9), 1-ethyl malate (10), 1-buthyl malate (11), succinic acid (12), ethyl-O-β-D-glucopyranoside (13), 1α, 2α, 4β-trihydroxy-1, 2, 3, 4-tetrahydronaphthalene (14), and L-2-O-methyl-chiroinosicol (15). Conclusion: Compounds 5 and 10-14 are isolated from the green walnut husks of J. regia for the first time, and compounds 1, 4, and 15 are isolated from this plant for the first time.
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PURPOSE: Walnut is known to have unique favorable fatty acids, phytochemicals, and other nutrient profiles. As a result, there has been growing interest in evaluation of its health benefit related to cardiovascular disease (CVD). Although inverse associations of nut consumption and risk factors of cardiovascular disease have been reported in many epidemiological studies and qualitative reviews, few meta-analysis studies have been reported. This meta-analysis was conducted in order to evaluate the effect of a walnut-enhanced diet on CVD risk factors. METHODS: We searched Pubmed, Cochrane, Science Direct, and KISS (Korean studies Information Service System) through July 2014. A random-effects meta-analysis was conducted on 17 trials reporting total cholesterol (TC), 14 trials reporting LDL cholesterol (LDL-C), 15 trials reporting HDL cholesterol (HDL-C), 17 trials reporting triglyceride (TG), and four trials reporting flow-mediated dilation (FMD). RESULTS: In meta-analysis, intake of a walnut-enhanced diet resulted in significantly lowered TC, LDL-C, and TG by -0.124 mmol/l (95% CI, -0.209, -0.039; p = 0.004), -0.085 mmol/lL (95% CI, -0.167, -0.004; p = l0.039), and -0.080 mmol/l (95% CI, -0.155, -0.004; p = 0.039), respectively. The overall pooled estimate of the effect on FMD was +1.313% (95% CI, 0.744, 1.882, p = 0.000). HDL-C was not affected by walnut intake. No statistical heterogeneity was observed for any analysis. Results of funnel plots and Egger's regression suggested a low likelihood of publication bias in all biomarkers (p > 0.05). CONCLUSION: Findings of this meta-analysis provide consistent evidence that walnut-enhanced diet intake reduces the CVD risk factors.
Subject(s)
Biomarkers , Cardiovascular Diseases , Cholesterol , Cholesterol, HDL , Cholesterol, LDL , Diet , Fatty Acids , Information Services , Insurance Benefits , Juglans , Nuts , Phytochemicals , Population Characteristics , Publication Bias , Risk Factors , TriglyceridesABSTRACT
Objective: To investigate the purification technology of total flavonoids from the diaphragma of Juglans regia by macroporous resin. Methods: Twelve macroporous resins were chosn with static and dynamic adsorption and desorption experiments to optimize the purification parameters. Results: AB-8 macroporous resin was found to have good adsorption and desorption effects. The optimal purification conditions were pH value of 4.70, sample mass concentration of 0.713 5 mg/mL, the loaded amount of 4 mL/g, and loading flow rate of 1.5 BV/h. The sample was eluted by water with 2 BV and 50% ethanol of 2.5 BV, respectively. The purity of total flavonoids increased to 72.25% after the purification, and the yield was 93.94%. Conclusion: AB-8 is an ideal resin with the best enrichment for separating and purifying the total flavonoids from diaphragma of J. regia.
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The aim of our study was to evaluate the gastro protective effect of aqueous extract of Juglans regia.L leaves in albino rats. Albino rats of wistar variety weighing 140-165gms were used in the experiment. The sexes were evenly divided into different treatment groups. The aqueous leaf extract of Juglans regia.L was investigated for its anti- ulcer activity against pylorus ligation, aspirin induced and ethanol induced gastric ulcer in rats at 500mg/kg body weight p.o. Histopathological assessment of rat stomach was carried out. A significant reduction (p<0.01) in ulcer index was seen in leaf extracts of Juglans regia.L treated rats of pylorus ligation, aspirin induced and ethanol induced gastric ulcer models. The gastro protective effect was further confirmed by histopathological examination of rat stomach. Thus the present study concludes the Juglans regia.L leaf extract having potential gastro protective effect in the three models tested.
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The proximate composition of eleven walnut (Juglans regia L.) genotypes (28 ŞK 010, 28 ŞK 055, 28 ŞK 041, 28 ŞK 601, 28 ŞK 925, 28 ŞK 028, 28 ŞK 118, 28 ŞK 350, 28 ŞK 930, 28 ŞK 850, 28 ŞK 036) and three walnut cultivars (Şebin, Bilecik, Kaman 1) produced in Turkey were determined. The oil content of the samples ranged from 61.32 to 69.35%, corresponding to an energy value of approximately 710 kcal per 100 g of kernel. The protein content ranged from 10.58 to 18.19%, and the carbohydrate composition was between 9.05 and 18.92%. The ash content ranged from 1.53 to 1.99%, and the moisture content of the kernels was between 1.91 and 4.48% the oleic acid content of the oils ranged from 17.90 to 33.35% of the total fatty acids. The linoleic acid content ranged from 43.15 to 60.20%. The linolenic acid content ranged from 9.98 to 13.00%. The palmitic acid content was between 5.21 and 8.40%. Stearic acid ranged from 2.36 to 4.25%. Potassium was the major mineral in all the samples, ranging from 359.73 to 482.97 mg/100 g. Calcium was the next most abundant mineral, ranging from 109.45 to 335.97 mg/100 g, followed by magnesium, ranging from 126.01 to 165.15 mg/100 g.
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Antidepressant, anti-inflammatory, antihypoxic and antioxidant activities of methanol extract of Juglans regia L., Juglandaceae, flower were investigated. Antidepressant activity was examined by forced swimming test and tail suspension test in mice. Antihypoxic activity was investigated in haemic and circulatory models. The effects were pronounced in both models. It produced statistically significant anti-inflammatory activity in carrageenan induced edema at nearly all doses, compared to control groups. IC50 for DPPH radical-scavenging activity was 674±27.6 µg mL-1. Extract showed good Fe2+ chelating ability (IC50 43±1.5 µg mL-1). It exhibited low antioxidant activity in linoleic acid peroxidation test. Its pharmacological effects may be attributed, in part, to the presence of phenols and ISSN 0102-695X flavonoids in the extract.
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The aim of this work was to study the influence of a walnut (Juglans regia) extract on the growth of Escherichia coli (E. coli) AB1157, on the plasmid DNA topology and on the labeling of blood constituents with technetium-99m (99mTc). An E. coli AB1157 culture, in stationary phase, was incubated with walnut and the growth of the culture was evaluated by optical density at 600 nm for 7 hours. Plasmid DNA samples were incubated with SnCl2 in presence or absence of walnut for 40 minutes, 0.8 percent agarose gel electrophoresis was performed, the gel was stained and the plasmid topological forms were visualized. Blood samples from Wistar rats were incubated with walnut extract and an assay of labeling of blood constituents with technetium-99m (99mTc) was performed. Blood cells and plasma were separated. The radioactivity in each fraction was counted and percentage of incorporated radioactivity ( percentATI) was determined. The results presented an inhibitory action of the growth of the E. coli AB1157 culture, no protective action of the walnut extract in plasmid DNA treated with SnCl2. Moreover, walnut was also not capable to induce modifications in the DNA mobility in agarose gel but walnut was capable to decrease the distribution of 99mTc on the blood cell compartment. In conclusion, our experimental data suggest that in the walnut extract has substances with an effect on the growth of E. coli culture, a potential action to increase the SnCl2 effect on plasmid DNA and also is capable to alter the distribution of 99mTc on the blood cell compartment probably due to redoxi properties.
O objetivo desse trabalho foi estudar a influência de um extrato de nogueira (Juglans regia) no crescimento de Escherichia coli (E. coli) AB1157, na topologia do DNA plasmidial e na marcação de constituintes sanguíneos com tecnécio-99m (99mTc). Uma cultura de E. coli AB1157, em faseestacionária, foi incubada com nogueira e o crescimento da cultura foi avaliado por densidade óptica a 600nm por 7 horas. Amostras de DNA plasmidial foram incubadas com SnCl2 napresença ou ausência de nogueira por 40 minutos, a eletroforese em agarose 0.8% foi realizada, o gel foi corado e as formas topológicas do plasmídioforam visualizadas. Amostras de sangue de ratos Wistar foram incubadas com extrato de nogueira e um ensaio de marcação de constituintes sanguíneos com tecnécio-99m (99mTc) foirealizado. Células sanguíneas e plasma foram separadas. A radioatividade em cada fração foi contada e a percentagem de radioatividade incorporada (%ATI) foi determinada. Os resultados apresentaram uma ação inibitória docrescimento da cultura de E. coli AB1157, nenhuma ação protetora do extrato de nogueira em DNA plasmidial tratado com SnCl2. Além disso,na nogueira também não foi capaz de induzir modificações na mobilidade do DNA em gel de agarose, mas a nogueira foi capaz de diminuir a distribuição de 99mTc no compartimento sanguíneocelular. Concluindo, nosso resultado experimental sugere que no extrato de nogueira existem substâncias com um efeito no crescimento de cultura de E. coli, uma ação capaz de aumentar oefeito do SnCl2 no DNA plasmidial e também ser capaz de alterar a distribuição de 99mTc no compartimento sanguíneo celular provavelmentedevido a propriedades redoxi.
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OBJECIVE:To study the preparation technology of Psoralea corylifolia-Juglans regia tincture.METHODS: Taking Psoralea corylifolia and Juglans regia as raw material,which were extracted respectively with ethanol and mixed after having been condensed,then added with surface-active agent to prepare the compound preparation Psoralea corylifolia-Juglans regia tincture.RESULTS: The tincture was bright and uniformly sable in color.CONCLUSION: The preformulation and the preparation technology are both reasonable and the tincture is stable in quality.
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A method for the infrared spectrophotometric determination of total ph-ospholipids in seed of Juglans regia and acinus of Citrullus vulgaris was reported. The average recovery of total phospholipid in this test was 97.2%, the CV was 1.66%. The contents of total phospholipid in seed of Juglans regia and acinus of Citrullus vulgaris were 438.46-528.85mg% and 241.78 -247.47mg% respectively. The rapid quantitative analysis of the distribution of various phospholipid components in these two specimens was carried out by using thin layer chromatographic scanning and the corrective method of absorbance proportional coefficient. The results showed that the main phospholipid components in both specimens were PC and PE. The fatty acid composition of the total phospholipids of both specimens were analysed by GC-MS-DS (gas chromatography-mass spectrometry-data system).