Effect of poria cocos on gastrointestinal motility in mice / 公共卫生与预防医学

Objective To investigate the regulatory effect of poria cocos on gastrointestinal motility in mice. Methods A total of 130 Kunming mice were randomly divided into negative control group, low-dose and high-dose groups of raw poria cocos powder, low-dose and high-dose groups of cooked poria cocos powder, low-dose and high-dose groups of poria cocos surrogate culture powder, low-dose, medium-dose and high-dose groups of poria cocos water extract, and low-dose, medium-dose and high-dose groups of poria cocos alcohol extract, with 10 mice in each group. The animals were administered by gavage for 7 days, once a day. After the last administration, the intestinal propulsion function test and gastric solid emptying test were conducted to observe the regulating effect of poria cocos on gastrointestinal motility of mice. Results Compared with the negative control group, the small intestine propulsion rate in the low-dose group of poria cocos surrogate culture powder was significantly increased (P<0.01). Except the high-dose group of raw poria cocos powder, the other poria cocos groups had higher gastric residual rate (P<0.05). Conclusion Poria cocos does not promote intestinal propulsion of mice under normal physiological condition, but it can inhibit gastric empting and exert a moderating effect on gastrointestinal function in normal mice.

Cloning and Expression Analysis of Squalene Epoxidase Gene in Poria cocos / 中国实验方剂学杂志

ObjectiveTo clone squalene epoxidase （SE）， a potential key rate-limiting enzyme involved in the synthesis pathway of Poria cocos triterpenes， from P. cocos and analyze for bioinformatics and expression. MethodThe total RNA was extracted by the kit and reverse-transcribed to cDNA. Specific primers were designed， and the cDNA was used as a template for cloning the SE gene， which was analyzed for bioinformatics. The expression of P. cocos qualene epoxidase（PcSE） was examined by Real-time polymerase chain reaction（Real-time PCR） in P. coco Shenzhou No. 10， Xiangjing 28， and 5.78 strains. ResultThe full length of PcSE is 1 571 bp， containing four exons and three introns. The obtained CDS sequence is 1 413 bp， encoding 470 amino acids. This protein is a hydrophobic protein with no signal peptide structure and has two transmembrane structural domains with a FAD/NAD （P） binding domain and SE structural domain localized to the mitochondrial membrane and the plasma membrane. The homologous sequence alignment with fungi of the Poriferae family is 80.92%， and the phylogenetic tree shows that PcSE protein is most closely related to P. cocos from the US. The results of Real-time PCR showed that the PcSE was expressed in all three strains， with the highest expression in 5.78 strain， and there was no significant difference in PcSE expression among the three strains. ConclusionFor the first time， the PcSE gene was cloned and analyzed from P. cocos， providing a basis for further research on the function of PcSE and the analysis of P. cocos triterpene biosynthesis pathway.

Extracts of Poria cocos polysaccharides improves alcoholic liver disease in mice via CYP2E1 and NF-κB inflammatory pathways / 中国中药杂志

The present study investigated the effect of extract of Poria cocos polysaccharides(PCP) on cytochrome P450 2 E1(CYP2 E1) and nuclear factor κB(NF-κB) inflammatory signaling pathways in alcoholic liver disease(ALD) mice and explored its protective effect and mechanism. Sixty male C57 BL/6 N mice of SPF grade were randomly divided into a control group, a model group, a positive drug group(bifendate, 200 mg·kg~(-1)), and high-(200 mg·kg~(-1)) and low-dose(50 mg·kg~(-1)) PCP groups. Gao-binge mo-del was induced and the mice in each group were treated correspondingly. Liver morphological and pathological changes were observed and organ index was calculated. Serum levels of alanine aminotransferase(ALT) and aspartate aminotransferase(AST) were detected. Malondialdehyde(MDA) and superoxide dismutase(SOD) in liver tissues were detected by assay kits. The levels of interleukin-6(IL-6) and tumor necrosis factor-α(TNF-α) were detected by ELISA. The activation of macrophages was observed by immunofluorescence staining and protein expression of CYP2 E1, Toll-like receptor 4(TLR4), NF-κB p65, and phosphorylated NF-κB p65(p-NF-κB p65) were analyzed by Western blot. The ALD model was properly induced. Compared with the model group, the PCP groups significantly improved the pathological injury of liver tissues. Immunofluorescence staining revealed that compared with the model group, the groups with drug intervention showed decreased macrophages in liver tissues. Additionally, the PCP groups showed reduced ALT, AST, MDA, IL-6, and TNF-α(P<0.05), and potentiated activity of SOD(P<0.01). PCP extract has the protective effect against alcoholic liver injury in mice, and the underlying mechanism may be related to the regulation of the expression of CYP2 E1 and inhibition of TLR4/NF-κB inflammatory signaling pathway to reduce oxidative stress and inflammatory injury, thereby inhibiting the development of ALD.

Carbon dots derived from Poria cocos polysaccharide as an effective"on-off"fluorescence sensor for chromium(Ⅵ)detection / 药物分析学报

Chromium is a harmful contaminant showing mutagenicity and carcinogenicity.Therefore,detection of chromium requires the development of low-cost and high-sensitivity sensors.Herein,blue-fluorescent carbon quantum dots were synthesized by one-step hydrothermal method from alkali-soluble Poria cocos polysaccharide,which is green source,cheap and easy to obtain,and has no pharmacological ac-tivity due to low water solubility.These carbon quantum dots exhibit good fluorescence stability,water solubility,anti-interference and low cytotoxicity,and can be specifically combined with the detection of Cr(Ⅵ)to form a non-fluorescent complex that causes fluorescence quenching,so they can be used as a label-free nanosensor.High-sensitivity detection of Cr(Ⅵ)was achieved through internal filtering and static quenching effects.The fluorescence quenching degree of carbon dots fluorescent probe showed a good linear relationship with Cr(Ⅵ)concentration in the range of 1-100 μM.The linear equation was F0/F=0.9942+0.01472[Cr(Ⅵ)](R2=0.9922),and the detection limit can be as low as 0.25 μM(S/N=3),which has been successfully applied to Cr(Ⅵ)detection in actual water samples herein.

Study on the mechanism of Poria cocos polysaccharides on the regulation of gluconeogenesis in liver of type 2 diabetic mellitus model rats / 中国药房

OBJECTIVE To investigate the effect and mechanism of Poria cocos polysaccharides on the regulation of blood glucose in type 2 diabetes mellitus （T2DM）model rats by phosphatidylinositol 3-kinase（PI3K）/protein kinase B （Akt）/forked box transcription factor O 1（FoxO1）pathway. METHODS SD rats were randomly divided into blank control group （no modeling ，no administration），model group （modeling，no administration ），metformin group （modeling，200 mg/kg）and P. cocos polysaccharide low-dose，medium-dose and high-dose groups （modeling，100，200，400 mg/kg），8 in each group. Except for blank control group ， other groups were given high fat diet combined with streptozotocin to construct the model of T 2DM rats. At the same time ， administration groups were given relevant dose of medicine intragastrically ，and blank control group and model group were given constant volume of water intragastrically ，once a day ，for consecutive 42 days. During the experiment ，general condition and bodyweight of rats were observed every day ；fasting blood glucose （FBG）of rats were collected ，and oral glucose tolerance test were conducted and area under curve （AUC）was calculated the day before last administration. After last medication ，the heart ，liver， kidney organ index were calculated ；the levels of HbA 1c，TC，TG，MDA，SOD，GSH-Px and hepatic glycogen content were detected. HE staining was used to observe the pathological changes of liver and pancreatic tissue ，and the pathological grade score was calculated. Western blot assay was used to detect the protein expressions of p-PI 3K，p-Akt，p-FoxO1， PEPCK and G 6Pase in liver tissues. RESULTS Compared with blank control group ，the rats of model group suffered cc1965@163.com from polydipsia ，polyphagia and polyuria ；the body weight ， the levels of SOD and GSH-Px ，the protein expressions of p-PI 3K，p-Akt and p-FoxO 1 were significantly decreased （P＜0.05）；liver and kidney organ index ，blood glucose level at 0，0.5 and 2 hours after intragastric administration of glucose solution ，AUC， FBG，HbA1c，serum levels of MDA ，TC，TG and hepatic glycogen content ，liver and pancreatic pathological grade score ，the protein expressions of PEPCK and G 6Pase were all increased significantly （P＜0.05）. Compared with model group ，the general condition of rats in P. cocos polysaccharide groups were all improved ，and all of above indicators had been reversed to varying degrees. CONCLUSIONS P. cocos polysaccharide can downregulate protein expressions of PEPCK and G 6Pase which are key enzymes of gluconeogenesis ，inhibit hepatic gluconeogenesis ，effectively decrease blood glucose levels and regulate glucolipid metabolism in T 2DM model rats by weakening oxidative stress and upregulating PI 3K/Akt/FoxO1 pathway.

Cellular behavior of carbon dots derived from Poria cocos polysaccharide / 药学学报

The biological behavior of carbon dots, especially the mechanism of cellular uptake and intracellular distribution, is the basis of its biomedical applications. In this paper, blue fluorescent carbon quantum dots were synthesized by hydrothermal method with Poria cocos polysaccharide as raw material, and the specific biological behavior of carbon dots entering cells was explored to evaluate its biological activity. It was characterized by transmission electron microscopy, UV-vis absorption spectroscopy, fluorescence spectroscopy, Fourier transform infrared spectroscopy, X-ray diffraction and X-ray photoelectron spectroscopy. Two different cell lines, immunocytes-RAW264.7 cells (mouse mononuclear macrophages cells) and cancer cells-4T1 cells (mouse breast cancer cells), were used as the research objects to study the uptake kinetics, uptake pathway, distribution and efflux of polysaccharide carbon dots in cells. The results showed that the carbon dots have a size distribution of 2 to 10 nm, and the average size was 6.85 nm. The carbon dots were mainly composed of C, O and N elements, with abundant surface functional groups such as -OH, C=O, C-N and C=C, and the fluorescence quantum yield was 4.72%. Carbon dots enter cells in a certain concentration and time dependence. Different cell lines have different uptake pathways. RAW264.7 cells enter the cells mainly by macrophage-specific phagocytosis, and a small part of the endocytosis is mediated by caveolin, while 4T1 cells are mainly mediated by grid protein endocytosis and giant cell drinking process. In summary, the synthesized carbon dots have good fluorescence properties, low cytotoxicity and excellent biocompatibility, which can be used for cell imaging applications.

Suitable planting area of Poria cocos in Jinzhai county of Dabie Mountains region / 中国中药杂志

Dabie Mountain in Anhui province is a genuine producing area of Poria cocos, commonly known as Anling. Jinzhai county in Anhui province is a traditional producing area of P. cocos, and it is also a key county for poverty alleviation in Dabie Mountains. Poverty alleviation of traditional Chinese medicine producing area is an important measure to implement the major strategic deployment of the central government. The planting of P. cocos is helpful to promote the development of traditional Chinese medicine industry in Dabie Mountains and help poverty alleviation. P. cocos is a saprophytic fungus with special demands on soil and ecological environment, and its planting appears a scattered and irregular distribution. Traditional investigation methods are time-consuming and laborious, and the results are greatly influenced by subjective factors. In order to obtain the suitable planting area of P. cocos in Jinzhai county, according to the field survey, the research team has explored the regional, biological characteristics and cultivation methods of P. cocos in the county, and obtained the altitude distribution area suitable for the growth of P. cocos. Then, the MaxEnt niche model was used to analyze the relationship between ecological factors and distribution areas, and the potential distribution zoning of P. cocos in Jinzhai county was studied. Combined with the characteristics of P. cocos planting pattern, taking ZY-3 remote sensing image as the data source, the maximum likelihood method was used to extract the area that could be used for P. cocos cultivation in Jinzhai county, and the reason why artificial planting P. cocos was mainly distributed in the west of Jinzhai county was analyzed. The suitable regional classification of P. cocos in Jinzhai county was obtained by superposition of suitable altitude distribution area, MaxEnt analysis and area extracted from remote sensing image, which provided data support for the planting planning of P. cocos in Jinzhai county.

Study on mechanism of enhancing antioxidant activity in fermented Ziziphi Spinosae Semen with Poria cocos / 中国中药杂志

In this study, the antioxidant property changes in fermented Ziziphi Spinosae Semen(FZSS) with Poria cocos were analyzed by DPPH, ABTS and FRAP methods. Then the content determination of active ingredients and ~1H nuclear magnetic resonance(~1H-NMR) spectroscopy were also used to investigate the mechanism of FZSS with P. cocos in enhancing the antioxidant activity. The results showed that the content of active ingredients such as total phenols, total saponins and total polysaccharides were significantly increased during the fermentation time. The results of ~1H-NMR metabonomics showed that the contents of amino acids such as leucine, lysine, valine and alanine, nitrogen compounds such as creatine, creatinine, and betaine, and secondary metabolites, for instance, jujuboside A and spinosin were higher after fermentation, and above components showed positive correlation with antioxidant capacity in Pearson correlation analysis. Therefore, it was inferred that the enhancement of antioxidant activity of FZSS may be the result of the joint action of various chemical components. This study preliminarily clarified the mechanism of FZSS in enhancing the antioxidant activity, and provided new research ideas for the product development and utilization of FZSS.

Establishment of UPLC Fingerprint of Poria cocos Aqueous Extract and Study on Its Spectrum-effect Relationship with Sedative and Hypnotic Effect / 中国药房

OBJECTIVE：To establis h the UPLC fingerprint of Poria co cos aqueous extract ，and to investigate its relationship with sedative and hypnotic effect. METHODS ：Ten batches of P. cocos from different areas were extracted with water to obtain the aqueous extract. UPLC method was adopted. The determination was performed on Waters HSS-C 18 column with mobile phase consisted of 0.1％ phosphoric acid solution-acetonitrile-methanol （gradient elution ） at the flow rate of 0.4-0.2 mL/min. The detection wavelengths were set at 210 and 242 nm. The column temperature was 40 ℃，and sample size was 2 μL. The fingerprints of 10 batches of P. cocos aqueous extracts were established by using the Similarity Evaluation System of TCM Fingerprint （2012A version），and the common peaks were identified. The sedative and hypnotic effects of 10 batches of P. cocos aqueous extracts from different areas under the synergistic action of pentobarbital sodium were investigated by taking the sleeping rate ，sleep latency and sleep duration of mice as the single efficacy index. After data transformation of single efficacy index and total efficacy （single indexes calculated by analytic hierarchy process ），grey correlation analysis was used to analyze the correlation between the common peaks in fingerprint of P. cocos aqueous extract and the single efficacy index and total efficacy. RESULTS ：There were 24 common peaks in 10 batches of aqueous extract of P. cocos ， and 11 components were identified ， i.e. 16 α-hydroxydehydrotrametenolic acid （peak 6），16α-hydroxytrametendic acid （peak 7），poricoic acid B （peak 9），dehydrotumulosic acid（peak 10），poricoic acid A （peak 12），polyporenic acid C （peak 15），3-O-acetyl-16α-hydroxydehydrotrametenolic acid （peak 17），dehydropachymic acid （peak 20），pachymic acid （peak 21），dehydrotrametenolic acid （peak 22），dehydroeburicoic acid （peak 24）. Grey correlation analysis showed ，the correlation between 24 peaks and sleep duration was greater than 0.6（0.611 5- 0.811 8）；the correlation between 24 peaks and sleep latency was greater than 0.6（0.605 9-0.790 4），except for peaks 14，24 and 2；the correlation of 24 peaks between sleeping rate was greater than 0.6（0.606 4-0.721 6），except for peaks 23，19，17 and 5； the correlation of 24 peaks between total efficacy was greater than 0.6（0.619 0-0.781 2），except for peaks 2，5，19. The top 10 chromatographic peaks related to the total efficacy were peak 15（polyporenic acid C ），peak 16，peak 8，peak 11，peak 12 （poricoic acid A ）， peak 1， peak 7 （16 α-hydroxytrametendicacid）， peak 3， peak 9 （poricoic acid B ） and peak 20 （dehydropachymic acid ）. CONCLUSIONS ：UPLC fingerprint of P. cocos aqueous extract was established and 11 components were identified. Ten components such as polyporus acid C are closely related to the total efficacy of sedation and hypnosis ，which preliminarily reveal the material basis of the sedative and hypnotic effect of P. cocos .

Analysis of triterpenoic acids in different medicinal parts of Poria cocos (Schw.) Wolf using supercritical fluid chromatography / 药学学报

Qualitative and quantitative methods were used to establish the quality of different medicinal parts of Poria cocos (Poriae Cutis, rubra Poria, white Poria, Poria cum Radix Pini) by using ultra-performance convergence chromatography coupled with photo-diode array and quadrupole time-of-flight mass spectrometry (UPC2-PDA-Q-TOF/MSE). A total of 18 chromatographic peaks were detected from Poria cocos by UPC2-PDA. Principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) were used to compare the four medicinal parts. The results showed that there were significant differences in the components of different medicinal parts, and the main triterpenoic acids were poricoic acid A, poricoic acid B, dehydroeburicoic acid, and dehydrotrametenolic acid. When combined with the common active component polyporenic acid C, a method for determination of five triterpenoic acids in different parts of Poria cocos was established. These components could be separated within 15 min, and the amount of methanol was 3.63% of that of HPLC method. Taking the five triterpenoid acids as an index, the content of triterpenoid acids in different parts of Poria cocos from high to low were Poriae Cutis, rubra Poria, white Poria and Poria cum Radix Pini. The method is simple, rapid, and uses minimal solvent. The mobile phase of environment-friendly gas carbon dioxide has unique advantages in reducing environmental pollution, which can provide a basis for the development and standard formulation of Poria cocos and its related products.

Poricoic acid A induces apoptosis and autophagy in ovarian cancer via modulating the mTOR/p70s6k signaling axis

Due to the high mortality and rapid disease progression, ovarian cancer remains one of the most common malignancies threatening the health of women. The present study was conducted to explore the anticancer effects and the underlying mechanisms of poricoic acid A (PAA), the main components of Poria cocos, on ovarian cancer. We investigated the anticancer effects of different concentrations of PAA in the SKOV3 cell line. Cell viability and proliferation were examined by CCK-8 assay. Cellular migration and invasion were assessed by the scratch and Transwell migration assays, respectively. The effect of PPA on cell apoptosis was measured by flow cytometry and caspase-3/8/9 colorimetric assay. Western blot was performed to detect protein level changes related to apoptosis and mTOR signaling pathways. The in vivo anticancer effect of PAA was evaluated using xenograft tumorigenesis model in nude mice. Our results showed that PAA suppressed SKOV3 cellular viability, migration, and invasion in a dosage-dependent manner. Flow cytometry results demonstrated PAA treatment could induce SKOV3 cell apoptosis. In addition, increased ratio of LC3-II/LC3-I (a marker for autophagosome formation) was observed after PAA treatment, as well as inhibition of m-TOR and p70s6k phosphorylation. In nude mice, PAA treatment reduced the xenograft tumor weight by 70% (P<0.05). In conclusion, our data suggested that PAA induced apoptosis and autophagy in ovarian cancer via modulating the mTOR/p70s6k signaling axis.

Regulatory effect of polar extract of Poria cocos on neurotransmitter and circadian rhythm disorder in CUMS rats / 中草药

Objective: To explore the regulation of polar extract of Poria cocos on neurotransmitter and circadian rhythm metabolic pathway. Methods: Male SD rats were randomly divided into blank group, chronic unpredictable mild stress (CUMS) model group, positive drug venlafaxine hydrochloride group (35 mg/kg), and polar extract of P. cocos group (15 g/kg). Rats were modeled and ig administrated for 28 d simultaneously, and the body weight, sugar preference rate, and opening behavior were observed dynamically after administration. The levels of tryptophan (Trp), 5-hydroxytryptophan (5-HT), and 5-hydroxyindole acetic acid (5-HIAA), noradrenaline (NE), dopamine (DA), acetylcholiner (Ach), γ-aminobutyric acid (GABA), glutamic acid (Glu), the proportion of GABA/Glu in serum were measured by LC-MS. The expression of key genes in the circadian rhythm pathway in liver tissue was detected by using qRT-PCR. Results: Polar extracts of P. cocos significantly improved the abnormality of the sugar preference rate and opening behavior. And polar extracts of P. cocos significantly restored the levels of 5-HT, 5-HIAA, Trp, NE, Ach, GABA, the proportion of GABA/Glu in serum and regulated the gene expression levels of Arntl, Per1, Per2, Per3, and Nr1d1 in hepatocyte. Conclusion: The polar extract of P. cocos could improve neurotransmitters and circadian rhythm disorders in CUMS rats through enhancing the 5-HT metabolic pathway, regulating Ach-NE signaling interaction and the equilibrium of proportion of amino acids neurotransmitter and restoring the expression levels of Arntl, Per1, Per2, Per3, and Nr1d1 involved in the circadian rhythm.

Research progress on chemical composition and pharmacological effects of Poria cocos and predictive analysis on quality markers / 中草药

Poria cocos is a traditional Chinese medicine with homology of medicine and food in China. It has the effects of promoting diuresis, eliminating dampness, invigorating spleen, calming heart and so on. It is widely used in medicine, food and health care products. With the in-depth study of P. cocos, its triterpenes, polysaccharides and other major chemical components, as well as its wide range of pharmacological effects and application development research have attracted much attention. This paper systematically reviewed the chemical components and pharmacological effects of P. cocos, according to the concept of quality markers, the quality markers of P. cocos were predicted and analyzed from the aspects of the biosynthetic approach and specificity of chemical components, traditional medicinal efficacy, traditional medicinal properties, measurable components, different processing methods and so on, which provides a scientific basis for quality evaluation and product development of P. cocos.

Prescription regularity of traditional Chinese medicine for treating alcoholic liver disease based on traditional Chinese medicine inheritance support system / 中草药

Objective: To summarize the prescription rules of traditional Chinese medicine (TCM) for treating alcoholic liver disease (ALD) based on TCM inheritance support system (V2.50). Methods: The literatures about TCM prescriptions for treating ALD were collected from CNKI, Wanfang database, and VIP database. The TCM inheritance platform system was used to analyze the prescription rules of TCM in the treatment of alcoholic liver disease. Results: Statistics showed that the majority of prescriptions were used to treat alcoholic fatty liver, alcoholic hepatitis, and alcoholic cirrhosis. Through "frequency statistics" analysis, 107 prescriptions were found involving 149 flavors of TCM, with a cumulative frequency of 1 195 times. Twenty-three Chinese medicines with a frequency of ≥ 15 times were used, and the cumulative frequency was 737 times (62%). The most frequently used medicines were blood-activating and stasis-removing drugs, water-diffusing and damp-permeating drugs, tonics, heat-clearing drugs, antialcoholic poisons and qi-regulating drugs. The commonly used doses of Salvia miltiorrhiza, Poria cocos, Bupleurum chinense, Glycyrrhiza uralensis, Atractylodes macrocephala, Alismatis Rhizoma, and Curcumae Radix in the top 10 medicines ranked in the frequency of medication accorded with the prescribed doses in the Pharmacopoeia of the People's Republic of China (2015 edition), while Crataegi Fructus, Artemisiae Scopariae Herba, and Puerariae Lobatae Radix exceeded the prescribed doses. In the frequency analysis of drug pairs, the combination of S.miltiorrhiza and B. chinense was the most widely used. According to the association rules of drug combination, the correlation between Curcumae Radix and S. miltiorrhiza was the strongest, that was, the probability of S. miltiorrhiza appearing with the emergence of Curcumae Radix was 88%. From the network display chart, it was indicated that S. miltiorrhiza and P. cocos were the main herbs for treatment. Through unsupervised entropy hierarchical clustering algorithm, 14 core combinations for new clustering were extracted, and seven new prescriptions can be obtained by further clustering. Conclusion: The basic principles of TCM treatment of ALD include promoting blood circulation and removing blood stasis, removing dampness, tonifying, detoxifying alcohol, and promoting qi, and with "protecting spleen and stomach function" as its purpose, which accords with the theoretical basis of traditional Chinese medicine in treating alcoholic liver disease. Core combinations and new prescriptions provide references for clinical drug use and new drug research and development, but new prescriptions must be further evaluated with the combination of traditional Chinese medicine theory and clinical practice.

Substance basis of warm nature of Poria cocos / 中草药·英文版

Objective: The research indicated that the nature of Chinese medicine is mainly related to body's substance and energy metabolism. The purpose of the study is to elucidate the substance basis for warm nature of Poria cocos (called Fuling (FL) in Chinese). Methods: In terms of the effects of its separated fractions on the substance and energy metabolism in rat models of cold-deficiency with Aconiti Lateralis Radix Praeparata (called Fuzi (FZ) in Chinese), with hot nature, as reference drug. Biochemical indexes in the material metabolism, energy metabolism, endocrine system, nervous system and nucleotide system were determined, then analyzed by additive, cluster and principal component analysis (PCA). Results: The medicinal natures of oligosaccharides and amino acids fractions were attributable to plain and crude polysaccharides, volatile oils and triterpenoids fractions were attributable to mild warm. Conclusion: The nature of FL was regarded as mild warm based on the old records of Chinese medicine and fractions of crude polysaccharides, volatile oils and triterpenoids might be the main substance basis for the warm nature of FL. It is the first time that substance basis of FL was elucidated from view point of medicinal nature.

Content Determination and Cluster Analysis of 7 Kinds of Triterpenes in Poria cocos from Different Habitats / 中国药房

OBJECTIVE：To establish a method for th e content determination of 7 kinds of triterpenoids in Poria cocos ，and to compare the differences of the above components in P. cocos from different habitats ，so as to provide reference for the quality control of P. cocos . METHODS ：Using 36 batches of P. cocos from different habitats as samples ，HPLC method was used for content determination of dehydrotomorphic acid ， polyporhinic acid C ， 3-epidehydrotomorphic acid ， 3-O-acetyl-16 α-hydroxy-hydrogenolysaccharic acid ，dehydrotomorphic acid ，pachymic acid and dehydrotrametenolic acid. The column was performed on Thermo Acclaim 120 C18 with mobile phase consisted of acetonitrile-phosphoric acid water （gradient elution ）at the flow rate of 1 mL/min. The detection wavelength was set at 210 nm. The column temperature was 30 ℃，and the injection volume was 20 μL. SPSS 21.0 statistical software was used for cluster analysis of 36 batches of P. cocos from different habitats. RESULTS ： The linearity of 7 triterpenoids was good in the range of their mass concentration （all r≥0.999 0）；average recoveries were 96.74％-104.04％（RSD＝0.54％-1.55％，n＝6）. RSDs of precision ，and reproducibility stability （24 h）tests were all lower than 3.0％（n＝6）. RSD of durability test was lower than 5.0％（n＝2）. There were some differences in the single content of 7 indicator components among samples from different habitats ，but the total content difference was not obvious （the total content of most samples was in the range of 1.3-1.9 mg/g）. After cluster analysis ，36 batches of sample were clustered into 5 categories，i.e. S 27 was clustered into class Ⅰ；S30 and S 34 were clustered into class Ⅱ；S2，S8 and S 9 were clustered into class Ⅲ；S10，S11，S12 and S 14 were clustered into class Ⅳ；and the remaining 26 batches of samples were clustered into class Ⅴ. CONCLUSIONS ：The method is simple ，and has good precision ，repeatability and durability. It can be used for the simultaneous determination of above 7 components in P. cocos . There has no significant difference in the quality of P. cocos from different habitats. The content of P. cocos in most producing areas is uniform in content and stable in quality ，only a few of them are different. Δ 基金项目 ：国家重点研发计划中医药现代化研究重点专项

Comparison of Total Protein Content Difference of Poria cocos from Different Producing Areas / 中国药房

OBJECTIVE：Compare the difference of total protein content of Poria cocos from different producing areas. METHODs：Using bovine serum albumin （BSA） as control ，0.4 mol/L sodium hydroxide solution as exctraction solution ， Coomassie brilliant blue G- 250 as chromogenic reagent ，visible spectrophotometry at 595 nm was used to determine the contents of total protein of P. cocos ；cluster analysis was used to classify 34 batches（S1-S34）of P. cocos from different producing areas. RESULTS：The linear range of BSA was 1.45-17.40 μ g/mL （r＝0.999 6）. RSDs of precision ，stability （20 min） and reproducibility tests were all lower than 3％；recoveries were 100.14％-104.26％（RSD＝1.43％，n＝9）. The contents of total protein in 34 batches of P. cocos from different producing areas were 0.388 4％-1.129 7％. The results of cluster analysis showed that among 34 batches of P. cocos ，the total protein content of P. cocos produced in Yingshan county of Hubei province （S2，S3） was higher than 1％，clustered into one category ；the total protein contents of P. cocos produced in Hubei ，Yunnan，Anhui and Hunan（S1，S5-S10，S12，S13，S16，S17，S19-S21，S23-S25，S28，S30，S31）were 0.653 5％-0.946 1％，clustered into one categony，and the remaining batch content were 0.388 4％-0.601 2％，clustered into one category. CONCLUSIONS ：Established method is suitable for the content determination of total protein content of P. cocos . The protein content of P. cocos from Yingshan county of Hubei province is the highest ，followed by Yunnan and Anhui in 34 batches of P. cocos from different producing areas.

Quality evaluation of Poria based on specific chromatogram and quantitative analysis of multicomponents / 中国中药杂志

HPLC specific chromatograms of Poria were established, and the concentrations of 10 triterpenoids(16α-hydroxydehydrotrametenolic acid, poricoic acid B, dehydrotumulosic acid, poricoic acid A, polyporenic acid C, poricoic acid AM, 3-O-acetyl-16α-hydroxydehydrotrametenolic acid, dehydropachymic acid, pachymic acid, and dehydrotrametenolic acid) were simultaneously determined. Chromatographic analysis was conducted on a Welch Ultimate XB C_(18) column(4.6 mm × 250 mm,5 μm). Acetonitrile solution(contain 3% tetrahydrofuran)(A) and 0.1% formic acid aqueous solution(B) were used as the mobile phase with gradient elution at a flow rate of 1.0 mL·min~(-1). The column temperature was 30 ℃ and the injection volume was 20 μL. The experimental data were analyzed by the SPSS 22.0 and GraphPad Prism 7.0. The established triterpenoids fingerprints were specific, and the 10 components were well separated and showed good linearity(r≥0.999 6) within the concentration ranges tested. The mean recoveries were between 98.53%-103.8%(RSD 1.7%-2.7%). The method was specific and repeatable, and could be used for identification and quality evaluation of Poria. The results showed that the contents of 10 triterpenoids were positively correlated with each other. The contents of 10 triterpenoids of samples collected from producing areas were higher than that collected from markets. The total contents of 10 triterpenoids of samples collected from Hubei and Yunnan province were slightly higher than that from Anhui province, but the contents of samples from Anhui province were varied in smaller ranges.

Isolation of homogeneous polysaccharide from Poria cocos and effect of its sulfated derivatives on migration of human breast cancer MDA-MB-231 cells / 中国中药杂志

SATB1 plays a crucial role in the invasion and metastasis of breast cancer,and inhibition of SATB1 expression can effectively control breast cancer metastasis. In this study,homogeneous polysaccharides were isolated from Poria cocos and their sulfated derivatives were prepared to screen out the polysaccharide compositions with inhibitory effects on SATB1 expression. Smal-molecule components were removed from P. cocos by ethanol extraction,and P. cocos crude polysaccharide PPS was obtained by water extraction and ethanol precipitation. Then PPS was successively separated by DEAE Sepharose fast flow anion-exchange and Superdex-75 gel permeation chromatographic steps to give PPSW-1. The structure of PPSW-1 was identified and its sulfated derivatives were prepared. Then their inhibitory effects on human breast cancer MDA-MB-231 cells were investigated. A kind of polysaccharide,PPSW-1 with inhibitory effect on human breast cancer MDA-MB-231 cells,was obtained from P. cocos,with a relative molecular weight of 3. 06×104,and structure of 1,6-branched 1,3-α-D-galactan. PPSW-1 and its sulfated derivative Sul-W-1 showed good inhibitory effect on cells migration,and the water solubility of Sul-W-1 was better than that of PPSW-1. In addition,it was found that polysaccharide of P. cocos and its sulfated derivative can inhibit expression of SATB1. In this study,a kind of homogeneous polysaccharide with inhibitory effect on human breast cancer MDA-MB-231 cells was isolated from P. cocos,and its sulfated derivative with similar efficacy but better solubility was prepared,laying the foundation for the substance basis study of P. cocos.

Non-polysaccharide chemical constituents of Poria cocos and their anti-complementary activity / 中草药

Objective: To investigate the non-polysaccharide chemical constituents of Poria cocos and their anti-complementary activity. Methods: The anti-complementary bioassay-guided isolation was carried out with the hemolysis test as guide. All isolates were evaluated for their in vitro anti-complementary activities on the classical pathway. The structures were identified by various spectroscopic data including ESI-MS, 1H-NMR, and 13C-NMR data. Results: Eleven compounds were isolated from the EtOAc fraction of P. cocos extracts, including stigmasterol (1), lupeol (2), oleanolic acid (3), ursolic acid (4), polyporenic acid C (5), tumulosic acid (6), dehydrotumulosic acid (7), 3-epi-dehydrotumulosic acid (8), pachymic acid (9), dehydropachymic acid (10), and dehydrotrametenolic acid (11). Compounds 1-4 were obtained from this plant for the first time, and compounds 3-11 showed the anti-complementary activity in different degrees. Conclusion: Triterpenoid acids are the main anti-complementary constituents in the chemical constituents of P. cocos non-polysaccharides (CH50 0.10-0.27 g/L).