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In the digital transformation of Chinese pharmaceutical industry, how to efficiently govern and analyze industrial data and excavate the valuable information contained therein to guide the production of drug products has always been a research hotspot and application difficulty. Generally, the Chinese pharmaceutical technique is relatively extensive, and the consistency of drug quality needs to be improved. To address this problem, we proposed an optimization method combining advanced calculation tools(e.g., Bayesian network, convolutional neural network, and Pareto multi-objective optimization algorithm) with lean six sigma tools(e.g., Shewhart control chart and process performance index) to dig deeply into historical industrial data and guide the continuous improvement of pharmaceutical processes. Further, we employed this strategy to optimize the manufacturing process of sporoderm-removal Ganoderma lucidum spore powder. After optimization, we preliminarily obtained the possible interval combination of critical parameters to ensure the P_(pk) values of the critical quality properties including moisture, fineness, crude polysaccharide, and total triterpenes of the sporoderm-removal G. lucidum spore powder to be no less than 1.33. The results indicate that the proposed strategy has an industrial application value.
Subject(s)
Bayes Theorem , Data Mining , Drug Industry , Powders , Reishi , Spores, FungalABSTRACT
Gemcitabine (GEM) is a commonly used drug in the clinical treatment of non-small cell lung cancer. Due to the accumulation of cells mediating immune escape and T cell depletion after chemotherapy, tumor microenvironment (TME) tends to be immunosuppressive status, which ultimately leads to tumor metastasis. The experimental protocol was approved by the Medical Laboratory Animal Ethics Committee of Jiangsu Provincial Academy of Chinese Medicine. Therefore, we observed the immunomodulatory effects of micro-particulate Ganoderma lucidum spore β-glucan (PGSG) on macrophages in vitro experiments. Next, mice subcutaneous Lewis lung cancer models were established to observe the anti-tumor effects of PGSG through oral administration of PGSG combined with GEM. Flow cytometry analysis was used to analyze the ratio of anti-tumor T cells in tumors and spleen, as well as the proportion of myeloid-derived suppressor cells (MDSC), tumor-associated macrophages (TAM) and regulatory cells (Tregs). The results showed that PGSG can up-regulate the expression of major histocompatibility antigens (MHC-II), CD40, CD86 and CD80 on the surface of macrophages, enhance the ability to phagocytosis of neutral red and further mediate the release of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), interleukin-4 (IL-4) and interleukin-10 (IL-10). In vivo experiments, combined administration can significantly decrease the volume and weight of tumors, reduce the ratio of MDSC (CD11b+Gr-1+), M-MDSC (CD11b+Ly6G-Ly6Chigh) and Treg (CD4+Foxp3+). At the same time, PGSG promoted the conversion of M2 (F4/80+CD206+) to M1 (F4/80+MHC-II+) and enhanced the response of helper T cell-1 (Th1) (CD4+IFN-γ+) and cytotoxic T lymphocyte (CTL) (CD8+IFN-γ+), which is of great significance for killing tumors. These results suggest that PGSG can regulate innate and adaptive antitumor immune responses, reshape the immunosuppressive microenvironment and enhance the anti-lung cancer effect of GEM.
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OBJECTIVE@#Ganoderma lucidum spore (GLS) is gaining recognition as a medicinal part of G. lucidum and has been reported to possess various pharmacological properties, such as antitumor activity. In this work, wall-broken GLS powder (BGLSP) and wall-removed GLS powder (RGLSP), two kinds of GLS powder with different manufacturing techniques, were compared in terms of contents of active constituents and in vivo and in vitro antitumor effects.@*METHODS@#The ultraviolet and visible spectrophotometry method was used to determine the contents of polysaccharides and total triterpenoids in BGLSP and RGLSP. Seventeen individual triterpenoids were further quantified using ultra-high-performance liquid chromatography and quantitative analysis of multi-components by single marker. The antitumor effects of BGLSP and RGLSP were evaluated using in vitro cell viability assay against human gastric carcinoma SGC-7901, lung carcinoma A549 and lymphoma Ramos and further validated by in vivo zebrafish xenograft models with transplanted SGC-7901, A549 and Ramos.@*RESULTS@#The results showed that the contents of polysaccharides, total triterpenoids and individual triterpenoids of RGLSP were significantly higher than those of BGLSP. Although both BGLSP and RGLSP inhibited the three tumor cell lines in vitro in a dose-dependent manner, the inhibitory effects of RGLSP were much better than those of BGLSP. In the in vivo zebrafish assay, RGLSP exhibited more potent inhibitory activities against tumors transplanted into the zebrafish compared with BGLSP, and the inhibition rates of RGLSP reached approximately 78%, 31% and 83% on SGC-7901, A549 and Ramos, respectively.@*CONCLUSION@#The results indicated that the antitumor effects of GLS were positively correlated with the contents of the polysaccharides and triterpenoids and demonstrated that the wall-removing manufacturing technique could significantly improve the levels of active constituents, and thereby enhance the antitumor activity.
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@#Models of acute and chronic liver injury in mice were established using carbon tetrachloride (CCl4) and ethanol to explore the protective effects of Ganoderma lucidum spore glycopeptide on liver injury.Different dosage of Ganoderma lucidum spore glycopeptide (65,130,260 mg/kg) were given by gavage.The liver index and the levels of serum aspartate transaminase (AST) and alanine transaminase (ALT) were determined.The contents of liver interleukin-6 (IL-6), tumor necrosis factor (TNF-α) and inducible nitric oxide synthase (iNOS) were tested by enzyme-linked immunosorbent assay (ELISA).The pathological injury of liver tissue was observed by HE staining.The results showed that Ganoderma lucidum spore glycopeptide could significantly reduce the liver index and the contents of serum AST and ALT in mice of acute and chronic liver injury.In mice of chronic liver injury induced by CCl4, Ganoderma lucidum spore glycopeptide could significantly decrease the contents of liver IL-6, TNF-α and iNOS, and alleviate the pathological damage of liver tissue.Results suggested that Ganoderma lucidum spore glycopeptide might reduce acute and chronic liver injury with anti-inflammatory effects in mice.
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@#To investigate the antitumor activity induced by Ganoderma lucidum spore oil on different tumor cells. Human hepatoma cells(HepG2), human non-small cells lung cancer cells(A549)and human colon cancer cells(HCT116)were selected and tested. Cell viability was determined by MTT assay. Western blot analysis was performed to measure the expression of NF-κB and Caspase-3 activity in order to elucidate the mechanism of apoptotic activity caused by Ganoderma lucidum spore oil and to screen out the most sensitive cancer cell lines to Ganoderma lucidum spore oil. MTT assay demonstrated that Ganoderma lucidum spore oil had a strong inhibitory effect on the growth of three cancer cell lines. Among these cells, A549 cells were most sensitive to Ganoderma lucidum spore oil, followed by HepG2 cells and then by HCT116 cells. The results of Western blot showed that Ganoderma lucidum spore oil could promote the activation of NF-κB pathway, and that the activation of NF-κB signaling pathway in cancer cells treated by Ganoderma lucidum spore oil was stronger in A549 cells, HepG2 cells, HCT116 cells respectively. The detection of Caspase-3 activity showed that ganoderma spore oil could activate Caspase-3 dependent apoptosis pathways, which was more important in A549 cells, HepG2 cells and HCT116 cells. This study found that Ganoderma lucidum spore oil had inhibitory effects on A549, HepG2 and HCT116 cells growth and that its antitumor activity was in time-dose dependence. The mechanism may be related to the activation of NF-κB pathway and the Caspase-3 apoptotic pathway, which could accelerate apoptosis and necrosis of tumor cells. Among the three kinds of cancer cells, A549 cells was most sensitive to Ganoderma lucidum spore oil, followed by the HepG2 cells, and then by HCT116 cells.
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Ganoderma lucidum is a kind of medicine and food fungi of genus Ganoderma, Polyporaceae. It’s regarded as traditional and precious medicinal material with high medicinal value in China. G. lucidum spore polysaccharide is the main active ingredient from G. lucidum. Now it is proved that it has wide medicinal value and wide application prospects in medicine. This article gives an overview of the research progress in the polysaccharide structure, extraction technology, and bioactivity of G. lucidum spore polysaccharides, which provides an important reference for the research and the medicinal value of natural products.
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OBJECTIVE: To develop a method for determination of monosaccharide composition of Ganoderma lucidum spore polysaccharide. METHODS: Ganoderma lucidum spore polysaccharide was extracted in boiling water and hydrolyzed by 2 mol · L-1 TFA. The monosaccharides were determined by ion chromatography on a CarboPac PA10 column, using 2.5 mmol · L-1 KOH as mobile phase. The column temperature was 30°C. The flow rate was 1.0 mL · min-1. RESULTS: In the certain range, the concentration of each monosaccharide had good linearity with its chromatographic peak area. The recoveries of monosaccharides ranged from 85.6% to 97.0% with RSDs less than 3%. The results showed that Ganoderma lucidum spore polysaccharide was composed of glucose, galactose, mannose, xylose, arabinose, fucose, etc. CONCLUSION: The proposed method has good separation effect and high sensitivity, which can be used for the determination of monosaccharide composition of Ganoderma lucidum spore polysaccharide.
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Objective To study the role of ganoderma lucidum spore oil and ganoderma lucidum extraction spore oil in neovascularization of human high malignant breast cancer .Methods Human high malignant breast cancer cell MDA-MB-231 and tumor-bearing nude mice established with MDA-MB-231 were treated with different doses of ganoderma lucidum spore oil and ganoderma lucidum extraction spore oil .Epidermal growth factor receptor ( EGFR) expression level was examined by Western blotting and the RNA expression levels of neovascularization related molecules such as EGFR , vascular endothelial growth factor (VEGF), metalloproteinases(MMPs), thrombospondin(TSP-1), platelet derived growth factor( PDGF) , fibroblast growth factor ( FGF) were detected by Real-time PCR.Results Both ganoderma lucidum spore oil and ganoderma lucidum extraction spore oil inhibited the expression of EGFR in vitro and in vivo in a dose-dependent way.Both compounds induced down-regulation of VEGF and up-regulation of TSP-1 at RNA level.The effect of Ganoderma lucidum extraction spore oil was more significant than that of ganoderma lucidum spore oil .Conclusion Both ganoderma lucidum spore oil and ganoderma lucidum extraction spore oil inhibite the expression of neovascularization related molecules and increase the expression of molecules inhibiting neovascularization , whereas the effect of ganoderma lucidum extraction spore oil is more obvious .
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Objective To observe the effect of Ganoderma lucidum spore powder on sugar tolerance and intestinal flora of diabetes rats induced by streptozotocin (STZ). Methods The diabetes model rats (divided into 2 groups: the therapy group and the control) were intragastric administration with equal volume of Ganoderma lucidum spore powder and distilled water for 10 weeks, respectively. Then the blood glucose level and intestinal flora amount were tested. Results The abnormal sugar tolerance of therapy group rats returned to normal and the amount of Bacillus bifidus in intestinal flora also recovered. There were significant differences compared with control. Conclusion Canoderma lucidum spore powder can decrease blood sugar level and adjusts intestinal flora amount of experiment rats.
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Objective:To explore the effects of Ganoderma Lucidum spore on the immunity of the immunosuppression mice with Glucocorticiods(GC).Methods:Different dosages of Ganoderma Lucidum spore were intragastriced to the immunosuppression mice with Glucocorticiods, and the weight of the immune organs and the phagocytic function of the macrophages and neutrophile,the expression in of CD3+,CD4+,CD8+,CD49b+ and CD19+ in the blood cell were measured;And the total complement activity, the content of the cytokine and the valence of hemolysin in the mice serum were detected.Results:The immune function of the GC immunosuppression mice were remarkable effected with Ganoderma Lucidum spore, it could significantly increase the index of the thymus and spleen, improve the valence of hemolysin and the phagocytic function of the macrophages and neutrophile of the mice.The total complement activity, the content of the cytokine in serum,and the quantity of T,B and NK cell in the mice blood were influenced by Ganoderma Lucidum spore.Conclusion:The results show that Ganoderma Lucidum spore remarkable effect on the immunity of the immunosuppression mice with GC, and it can promote accommodation the immune function of mouse.