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Use of fungal immunomodulatory protein

A technology of immunomodulatory proteins and uses, applied in peptide/protein components, medical raw materials derived from fungi, plant/algae/fungus/moss components, etc., can solve problems such as cell apoptosis and uninvestigated molecular mechanisms

Inactive Publication Date: 2007-04-04
YEASTERN BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

A triad of Ganoderma tsugae was found to induce apoptosis and cell cycle arrest in human liver cancer cells Hep3B, but the molecular mechanism has not been investigated (Gan KH et al., J Nat Prod 1998; 61(4 ): 485-487.)

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Example 1: Change cell morphology

[0064] The A549 cell line belongs to human lung cancer epidermal cells, and this lung cancer cell line is rather malignant due to its high migration capability. In the present invention, the A549 lung cancer cell line is used as a cancer model to study the response of cancer cells to FIP-gts.

[0065] First, the present invention observes the morphological changes of cells treated with FIP-gts under a microscope. A549 cells were treated with FIP-gts at different concentrations (0, 1, 2, 4 and 10 μg / ml) and observed at different times and photographed. The morphological changes of the cells were recorded (Figure 1).

[0066] A549 cells were treated with 2, 4 or 10 μg / ml of FIP-gts, and the cell morphology changed significantly after 6 hours. The cell morphology changed from a state of adhering with little tentacles to round and A round and loosly-attached morphology, these round cells will float with gentle shaking of the dish. Afte...

Embodiment 2

[0068] Example 2: Cell viability assay

[0069] To further confirm the above experimental results, the present invention uses trypan blue staining to check cell viability. In the present invention, the cells were treated with the same FIP-gts concentration (0, 1, 2, 4 and 10 μg / ml of FIP-gts) as the previous experiment for 48 hours, and then trypan blue was added, and the living cells would not be stained by trypan blue, In the present invention, the number of unstained cells (live cells) is calculated as the viability of cells.

[0070] The present invention was inoculated in 2 x 10 in 6 cm petri dishes 5 H1355 and A549 cells, H1355 cell line is a cell model for general research on metabolism. After culturing at 37°C for 16 hours, replace the new culture medium, and add different concentrations of FIP-gts (0, 1, 2, 4 and 10 μg / g / g / ml).

[0071]The FIP-gts drug-treated cells were collected after 48 hours, the old culture medium was removed and the cells were collected in a...

Embodiment 3

[0073] Example 3: Counting the number of cells - Colony forming

[0074] In this experiment, the cell colony forming method will be used to analyze the poisoning effect of FIP-gts on cancer cells. A549 cells were treated with different concentrations of FIP-gts (0, 0.4, 2 and 10 μg / ml) and FIP-gts drugs. After 24 hours, the treated cells were seeded with 400 cells per 6 cm culture dish for 12 days.

[0075] First, the present invention was individually inoculated in 2×10 6 cm petri dishes 5 After culturing A549 cells at 37°C for 16 hours, the culture medium was replaced with a new medium, and different concentrations of FIP-gts (0, 0.4, 1, 2 and 10 μg / ml) were added at the same time (Figure 4). After 24 hours of treatment, the cells were washed twice with 1×PBS, and the drug-treated cells were reinserted, and 1 ml of TE buffer solution was added and allowed to stand at 37° C. for 1 minute to break up the cells. Serial dilutions were used to accurately estimate the number of ...

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Abstract

This present invention relates to the use of fungal immunomodulatory protein in immunotherapy, treating or activating natural killer cells, macrophage or increasing cytokines and serum antibody.

Description

technical field [0001] The present invention relates to the use of fungal immunomodulatory proteins for immunotherapy and lowering blood sugar. Background technique [0002] Ganoderma lucidum (Genus Ganoderma) is a rare and precious Chinese medicinal material. It is commonly known as "Ganoderma lucidum" in China and has been used for 5,000 years. Ganoderma lucidum currently in use includes: G.lucidum (red), G.applanatum (brown), G. tsugae (reddish brown), G. sinense (black) and G.oregonense (dark brown). [0003] Ganoderma lucidum has anti-allergy (Chen H.Y et al., J. Med. Mycol. 1992; 33: 505-512), liver protection (Lin J. M. et al., Am J Chin Med. 1993; 21(1): 59-69 ), anti-cancer (Wasser SP, CritRev Immunol 1999. 19: 65-96) and functions such as increasing immunity (Kino, J Biol. Chem. 1989. 264(1): 472-8). However, Ganoderma lucidum is used by direct extraction (Horner W.E. et al., Allergy 1993; 48: 110-116) or by using extracted small molecules (Kawagishi H., et al., ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/16A61K36/06A61K36/064A61K35/74A61P37/04A61P3/10
CPCC07K14/375A61K38/16G01N2333/9125G01N2333/37G01N33/57407A61K36/06A61P3/10A61P35/00A61P35/04A61P37/02A61P37/04Y02A50/30
Inventor 陈子智柯俊良
Owner YEASTERN BIOTECH
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