Huaier mushroom in cancer therapy

Reading time 16 minutes

Updated - January 12, 2026

The Huaier mushroom has been known for over 1,600 years and has been used successfully in cancer therapy since its cultivation.

history

It was first mentioned in a medical work around 240 AD Zhou Hou Fang of the doctor Ge Hong. The title refers to the period after the Zhou dynasty. It translates as „Manual of Emergency Medicine“, which focuses on prescriptions for first aid and is still referenced in medical contexts today.
It can also be found in the book Tang Ben Cao (Tang Dynasty), which was regarded as a reference work on herbal medicine.

The Huaier mushroom (Trametes robiniophila Murr) was used to treat chronic ailments, to speed up convalescence and for general strengthening. Among other things, it was said to promote blood circulation and eliminate the symptoms of the root causes of tumors.
Due to a lack of sufficient availability - the mushroom only grew in remote areas on the trunks of old specimens of the Chinese Sophora tree - it soon fell into oblivion.

Towards the end of the 1970s, Chinese scientists developed a method of cultivating the medicinal mushroom. Standardized production with consistent active ingredient quality has been possible since the early 1990s.

The Polysaccharide protein-complex (PS-T), consisting of polysaccharides and proteins, is the main active ingredient: a combination of a 6 Monosaccharides existing Heteropolysaccharide and one from 18 Amino acids composite protein.

Manufacturer

The manufacturer of Huaier granules is the company founded in 1995. Gaitianli Medicine Co, Ltd. based in Qidong, Jiangsu. The company maintains research, development and production facilities, test laboratories and warehouses on its approximately 130,000 square meter site with 1,700 employees. The production capacity amounts to 9,500 tons of Huaier mushroom and 250 million bags of granules.

The research focus is on the treatment of tumor and immune diseases. In studies, Huaier granules have shown promising results for treating cancer and blocking recurrences (source).
The Huaier mushroom is considered a research focus, especially in breast cancer therapy.

The product was initially produced as a reference substance for clinical studies (32% polysaccharides and 8% β-glucans) and was finally officially approved in China as an adjuvant therapeutic agent in oncology.

Today it is sold via pharmacies (PZN 19253502 - only 30% polysaccharides) and online platforms (Nutrimentas granules (identical concentration to the study content with 32% polysaccharides)) are distributed worldwide.
The Nutrimentas granules follow the scientific standard of the original manufacturer Gaitianli Medicine Co., Ltd. (32% polysaccharides, 8% β-glucans)

Studies

The most recent study from 2024, which has since been confirmed several times and was first published in 2022, is currently the Tanaka study by Dr. Manami Tanaka, M.D., Ph. D., Kanagawa, Japan, who works in biomedical research. His very detailed work, on the occasion of which the modRNA-vaccinations (Corona) ribosomal RNA (rRNA) in relation to the effect of the Huaier mushroom showed, contrary to the actual intention of his work, that cancer also responds to Huaier.

The study shows that taking Huaier extract in cancer patients has several effects: it normalizes ribosomal function, reduces the production of harmful spike proteins and prevents cancer recurrence with continued use.

The Huaier fungus has an „all-round“ effect - with the exception of brain tumors, as here the molecules probably cannot pass the blood-brain barrier due to the molecular size of the Huaier active substances with TP-1: 2300 kDa, HP-1: 30 kDa* - and is not selectively limited to just a few types of cancer.
This is because the active ingredients merely ensure the functional normalization of cell functions. This sounds very succinct, but is highly complex, as the study clearly demonstrates.

*kDa is used to indicate the mass of molecules, especially proteins. The unit Dalton (Da) is defined as the twelfth part of the mass of the carbon isotope 12C and is 1.66053906660(50) - 10-²⁷ kg. kDa is practically identical to kg/mol.

Further studies:

• Immunoregulatory effects of Huaier (Trametes robiniophila Murr) and relevant clinical applications.
• An extraction from Trametes robiniophila Murr. (Huaier) inhibits non-small cell lung cancer proliferation via targeting to epidermal growth factor receptor.
• Polysaccharides Produced by the Mushroom Trametes robiniophila Murr Boosts the Sensitivity of Hepatoma Cells to Oxaliplatin via the miR-224-5p/ABCB1/P-gp Axis.

Function - explained for medical laypersons

Legal information: This information is for educational purposes and does not replace the advice of a specialist doctor/oncologist. Huaier granules are classified as a dietary supplement in Germany and not, as in China, as a medicinal product. Individual medical decisions should always be discussed with the treating oncologist.

Cancer cells undermine the body's control mechanism, the so-called. Hippo-Pathway (which determines whether a cell is healthy or defective and thus its cell death, the Apoptosis, ) and thus prevent apoptosis, which is why they continue to divide and multiply in an uncontrolled and uninhibited manner.

Huaier takes care of the repair of the Hippo-Pathways and thus enables the cell to resume its proper function, the switching on and off of various genes, and to correctly recognize and eliminate defective cells (Apoptosis).

To make matters worse, the so-called. Killer cells of the body (immune cells, NK cells) are exhausted in cancer patients and can no longer fulfill their task of destroying cancer cells.

Huaier activates these killer cells through his β-Glucans, This allows the immune system to be retrained and cancer cells such as metastases to be actively attacked and destroyed.

There are 8 main mechanisms by means of which the Huaier fungus successfully performs its amazing work, described below in excerpts and easy to understand.

1. the restoration of cell memory

A cell has specific tasks that are stored in the cell nucleus in the RNA (- RiboNuclein Acid = DNA - DeoxyriboNuclein Acid). A section of it codes for proteins that determine the function and structure of the cell by switching certain genes on or off.
To prevent this from happening, a gatekeeper (Hippo-Pathway) to ensure that the cell only fulfills its intended function. If it fails to do so, it is given around half an hour to correct the error. If it remains faulty, cell death (Apoptosis) is initiated to prevent a cell with incorrect information from multiplying.
However, if the gatekeeper fails, the incorrectly encoded cell will continue to divide inexorably.

The Huaier fungus reactivates the gatekeeper and thus restores control of cell function.

2. the genetic chaos

If the wrong genes are activated or deactivated or incorrectly switched on or off as a result of the incorrect information, other proteins are produced than required. The transcription factors are disrupted. As a result, the cell loses its assigned function.

However, genes are not simply switched on or off in binary form as in digital logic circuits, but are also finely adjusted, similar to a volume control, i.e. set to very quiet, quiet, medium, etc.. According to this setting, they ensure a reaction (expression) to the transmitted signals that is adapted to the respective situation.

The active ingredients of the Huaier mushroom reactivate thousands of genes correctly, bringing them back to their natural state, allowing the cell to resume its originally assigned function.

3. the metastasis brake

The signal paths PI3K, ACT and mTOR are used for intracellular communication, which determine growth, division behavior and metabolic processes, for example. If these are disrupted, the cell cannot develop its original function. If the signaling pathways are hyperactive, coordination of the processes is impossible, with the result that the cell goes out of control and divides rapidly (forms metastases).

Huaier inhibits this hyperactivity of the signaling pathways and thus prevents uncontrolled cell growth, including division and proliferation in the organism.

4. the miRNA control system

Compared to a car, the miRNA-control system (microRNA) represents the ABS control (spinning wheels are braked while gripping wheels are supplied with drive power). It prevents cells from skidding due to incorrect information.
Around 1,000 miRNAs are encoded in the human genome and control the switching state of genes.

For cancer Oncogenes (mutated genes that cause the growth of the cell to get out of control) are no longer slowed down, which promotes the growth and spread of the cancer.

At the same time Tumor suppressor genes (genes that control cell growth and division) are too strongly inhibited, which in turn allows the cancer to grow unchecked.

The Huaier fungus restores the defective genes to their intended switching position, which stops uncontrolled cell growth and prevents the defective cell from continuing to divide.

5. the exhausted immune system

The immune system is ultimately overwhelmed by the multiple malfunctions and can no longer adequately fight the cancer or keep it in check. The smallest infection, be it a cold, can develop into a full-blown pneumonia - with often fatal consequences.

The substances of the Huaier mushroom activate the NK cells (natural killer cells) and Macrophages (phagocytes), which kill cancer cells (Apoptosis) and absorb them in order to break them down. This allows the immune system to resume its protective function and fight the cancer effectively.

6 The wanderlust of cancer cells

Metastasis is the appearance of cancer cells in places other than the original tumor site. Normally, cancer cells adhere to the tissue where they originated. Over time, however, the EMT (Epithelial-mesenchymal transition) for the loss of the adhesive that holds the cell in place. As a result, it migrates around the organism, forming a movement protein that makes it even easier for it to move on.

The Huaier fungus inhibits this EMT process and thus prevents metastasis.

7 The supply system of cancer cells

Migrating cancer cells want to live and therefore form new blood vessels (Angiogenesis) to take care of themselves. This is how new cancerous tumors develop in various places in the body.

Huaier inhibits the growth factor VEGF, which is regulated by the hypoxia-inducible factor HIF-1α. This cuts off the supply to the resulting ulcers, which is why the tumor dies.

8. the defect in ribosomal RNA after chemotherapy

Chemotherapeutic agents damage the ribosomal RNA because they do not selectively target the DNA, but also affect other cellular structures.
Some chemotherapeutic agents, such as Actinomycin D, The DNA synthesis inhibitors, which are directly incorporated into the DNA double strand, not only prevent the formation of new DNA, but also the synthesis of RNA, including ribosomal RNA (rRNA), which is essential for protein production.
The cell thus loses its ability to synthesize proteins, which leads to cell death.

According to current research, the acute toxicity of Azacitidine mediated almost entirely via RNA damage. RNA damage apparently plays a central role in the effect of such drugs. Other substances, such as Anthracyclines, act by forming free radicals, which can damage both DNA and RNA.
This damage to the ribosomal RNA disrupts the function of the ribosomes, which are responsible for translation (Translation) of mRNA into proteins, which can ultimately lead to cell death.

The Huaier fungus repairs damage to the ribosomal structures, which helps healthy cells to regenerate, but causes cancer cells to die.

Active ingredients

The main active ingredients of the Huaier mushroom are divided into

1. β-glucans (beta-glucans) - 20-30% of the extract

• Polysaccharides with 1,3- and 1,6-glycosidic bonds
• Activate Toll-like Receptors (TLR2, TLR3, TLR6) on immune cells
• Stimulate Natural Killer Cells (NK cells) and Macrophages
• Increase TH1 cytokine production (IFN-γ, IL-2, TNF-α)

2. polysaccharides (30-40% of the extract in total)

• Modify the intestinal microbiota
• Promote the production of short-chain fatty acids (SCFAs)
• This activates G protein-coupled receptors (GPR43, GPR109A)
• Leads to epigenetic changes in immune cells

3. bioactive metabolites

• Polysaccharides with branched structure
• Triterpenes
• Phenolic compounds with antioxidant effect

When does taking Huaier granules have an effect?

The intake should be in direct connection with

• conventional surgery (accelerates wound healing)
• Chemotherapy (regenerates ribosomal RNA, prevents side effects)
• Irradiation
  (after prior discussion with the treating oncologist and his knowledge of these contents)
• hormone therapy, as there are no known interactions
• immunotherapy, due to synergistic effect

If the Huaier granules are taken regularly at the recommended dosage, the following effects can be observed:

Day 1-7:

• β-Glucans activate macrophages & NK cells
• First immune response is launched

Week 1-2:

• Transcription factors are reactivated
• First gene expression changes in cancer cells

Week 2-4:

• Massive gene expression conversion (1000s of genes)
• Hippo-Pathway is being repaired
• First apoptosis (cell death) in cancer cells

Week 4-12:

• EMT is blocked (metastasis prevention)
• Angiogenesis is inhibited (tumor starves)
• Immune system is completely retrained

Month 3+:

• Stable control of the remaining cancer cells
• Prevents recurrences and metastases
• Regenerate normal cells (especially after chemo)

Dosage recommendation

Using the example of metastatic breast cancer after resection and removal of 7 affected lymph nodes, the evidence-based dosage recommendation based on the Nutrimentas granules with 32% polysaccharides as follows:

Phase 1: Acute phase - after resection Weeks 1-4

Tumor burden: high (7 affected lymph nodes, risk of metastasis)

Recommended total daily amount: 60 g

• Divided: 3 × 20 g daily (morning, noon, evening)
• Time: best on an empty stomach or between meals

Active ingredient content in this phase:

• 60 g × 32% = 19.2 g polysaccharides
• Of which at least: 60 g × 28% = 16.8 g β-glucans

Preparation per dose:

1. Pour 20 g of granules into a cup
2. Pour approx. 100 ml of hot water (80°C) over it
3. Stir well until completely dissolved
4. Fill up to approx. 250 ml with lukewarm water
5. Drink slowly

Phase 2: Consolidation phase - weeks 5-12

After stabilization and first check-up

Recommended total daily amount: 30 g

• Divided: 3 × 10 g

Active ingredient content in this phase:

• 30 g × 32% = 9.6 g polysaccharides
• Of which at least: 30 g × 28% = 8.4 g β-glucans

This represents the standard dose in oncology and is used in most studies.

Phase 3: Maintenance phase - from the 4th month for a further 6-12 months

Recurrence prevention and metastasis prevention

Recommended total daily amount: 15 g

• 3 × 5 g daily = 15 g

Active ingredient content per day

• 15 g × 32% = 4.8 g polysaccharides
• Of which at least: 15 g × 28% = 4.2 g β-glucans

Important notes:

• Consistency is important: Daily intake without interruption is essential for optimum effect
• Continuous application: In order to ensure the therapeutic effects, the intake should be continued for at least 6-12 months
• Can be combined with conventional medicine: There are no known interactions
• Gentle on the stomach: Better tolerated if the granules are taken on an empty stomach
• Compatibility: Slight detoxification reactions may occur in the first 1-2 weeks (tiredness, headaches). These are normal and subside quickly.

Control examinations

Baseline - before taking Huaier

Blood tests:

• Tumor markers: CEA (carcinoembryonic antigen) - relevant for breast cancer
• Tumor markers: CA 15-3 (particularly important for breast cancer)
• Tumor markers: CA 27.29 (additionally for chest)
• Tumor markers: HER2/new (if not yet known)
• Full blood count: RBC, WBC, Hemoglobin, Hematocrit, Thrombocytes
• Liver function: AST, OLD, GGT, Bilirubin (important, as liver damage is possible with metastases)
• Kidney function: Creatinine, BUN, GFR
• Inflammatory markers: CRP, erythrocyte sedimentation (ISR)
• Immune function: Lymphocyte count (CD4, CD8, NK cells, if possible)

Tumor markers - specific interpretation in breast cancer

CEA (Carcinoembryonic antigen)

• Normal: < 2.5 ng/mL (< 5 ng/mL for smokers)
• What does increase mean? Relapse or metastatic disease
• Sensitivity: 50-70% for metastases

CA 15-3 (Cancer Antigen 15-3)

• Normal: < 25 U/mL (some labs < 35 U/mL)
• What does increase mean? Particularly relevant for metastatic breast cancer
• Sensitivity: 70-80% for metastases, only 25% for early stage

CA 27.29

• Normal: < 38 U/mL
• Which means: Breast cancer-specific marker
• Additional information on CA 15-3

Interpretation under Huaier:

• Good sign: Markers fall continuously or stabilize at a low level
• Warning signal: Continuous increase despite Huaier (= possibly Non-responder*)
• Remark: Individual measured values are not too important, the trends are decisive!

*Recognize non-responders

Warning signals indicate a lack of Huaier efficacy at this dosage:

• Tumor markers rise continuously (despite regular intake of Huaier)
• Lymphocytes remain low (< 20%)
• CT/MRI shows tumor progression
• New metastases on imaging
• Clinical deterioration (weight loss, loss of performance)

In this case, the daily dose of Huaier should be increased to 30-40g/day.

Signs of a positive effect

Blood laboratories:

• ✓ Tumor markers fall continuously
• ✓ Lymphocytes increase
• ✓ Normalization of liver and kidney function
• ✓ CRP (inflammation value) normalizes after initial increase

Imaging:

• ✓ Tumor regression or stabilization
• ✓ Lymph node reduction
• ✓ No new metastases

Clinical condition:

• ✓ Rising energy
• ✓ Improved appetite
• ✓ Better quality of sleep
• ✓ Psychological stabilization
• ✓ Hair growth (signal for stem cell activation)

Blood count parameters

Changes expected while taking Huaier:

Lymphocytes (normal: 20-40% of the WBC)

• Expected change: ↑ increase (= good sign, immune activation)
• Target: > 30%, ideally > 35%

Hemoglobin (normal: 12-16 g/dL in women)

• Expected change: ↑ Stabilization/slight increase
• Huaier supports blood formation (important after chemo)

Platelet count (normal: 150-400 K/μL)

• Expected change: ↑ Stabilization/increase
• Huaier also supports blood formation here

CRP (normal: < 3-5 mg/L)

• Expected change: ↑ Slight increase in week 1-2 (= immune response)
• Then ↓ decline in week 3-4 (= good sign)
• Shows immune activation

Imaging (baseline):

• CT thorax + abdomen (looking for lung metastases and hepatic metastases)
• Skeletal scintigraphy or PET-CT (searches for bone metastases)
• Locoregional assessment (surgical site, axillary lymph nodes)
• Optional: MRI liver (if hepatic involvement is suspected)

Phase 1: Acute phase - weeks 1-4

Dosage: 3 × 20 g daily = 60 g/day

Week 2

• Clinical assessment:
  • Tolerance, side effects, energy level
  • Appetite, sleep quality
  • Gastrointestinal tolerance (nausea, diarrhea)
• Laboratories (optional, only if available):
  • Rapid blood count (WBC, Lymphocytes)
  • CRP (inflammation)
  • Tumor markers (CEA, CA 15-3) - often still too early for significant change

Week 4

• Clinical assessment: General condition, wound healing (if recently operated on)
• Blood tests:
  • Tumor markers: CEA, CA 15-3, CA 27.29 (first response check)
  • Full blood count (WBC, Lymphocytes, Hemoglobin)
  • Liver function (AST, OLD, GGT, Bilirubin)
  • Kidney function (Creatinine, GFR)
  • CRP (inflammation marker)
  • If available: Lymphocyte profile (CD4/CD8 ratio, NK cell count)
• Notes
  • ✓ Tumor markers may still rise slightly in this phase (first „detoxification“)
  • ✓ Lymphocyte count often increased (immune activation)
  • ✓ CRP may be slightly elevated (immune reaction)

Phase 2 - Consolidation phase - weeks 5-12

Reduce dosage to: 3 × 10 g daily = 30 g/day

week 6

• Clinical assessment: Energy level, symptom complaints

Week 8

• Imaging:
  • CT thorax + abdomen (first image control)
  • Question: Size regression of primary tumor? New metastases? Lymph node regression?
  • Comparison with baseline
• Blood tests:
  • Tumor markers: CEA, CA 15-3, CA 27.29
  • Full blood count
  • Liver function
  • Kidney function
  • Immune markers (if available)
• Notes
  • ✓ Tumor markers should now start to fall (or be stable)
  • ✓ Imaging should show initial regression or stabilization
  • ✓ Lymphocyte count persistently elevated (good sign)

Week 12

• Blood tests:
  • Tumor markers (CEA, CA 15-3, CA 27.29)
  • Full blood count
  • Liver function
• Clinical assessment:
  • Decision for phase 3?
  • Responder vs. non-responder assessment

Phase 3 - Maintenance phase - from month 4 for 6-12 months

Dosage: 3 × 5 g daily = 15 g/day (or alternatively 2 × 5g = 10g/day)

Month 4 (week 16)

• Blood tests:
  • Tumor markers: CEA, CA 15-3, CA 27.29 (response assessment)
  • Full blood count
  • Liver function, kidney function
  • Immune markers
• Clinical assessment:
  • Overall evaluation of therapy success to date
  • Compatibility, quality of life
  • Possible adjustment of dosage based on markers

Month 6 after start

• Imaging (CRITICAL):
  • CT thorax + abdomen or PET-CT
  • Comparison with week 8 imaging and baseline
  • Goal: Confirmation of stable disease or further regression
• Blood tests:
  • Tumor markers (CEA, CA 15-3, CA 27.29)
  • Full blood count
  • Liver function, kidney function
  • CRP
  • Hormonal markers (if hormone therapy is planned)

Month 9

• Blood tests:
  • Tumor markers
  • Full blood count

Month 12

• Imaging (FOLLOW-UP):
  • CT thorax + abdomen or PET-CT
  • Assessment of long-term responses
  • Search for delayed metastases
• Blood tests (COMPLETE):
  • Tumor markers: CEA, CA 15-3, CA 27.29
  • Full blood count
  • Liver function, kidney function
  • CRP
  • Immune markers (if available)

Long-term monitoring from year 2

Dosage: 2 × 3-5 g daily = 6-10 g/day (maintenance)

Every 3 months:

• Blood tests: Tumor markers (CEA, CA 15-3, CA 27.29) + complete blood count

Every 6 months:

• CT or MRI (depending on the oncologist's protocol)
• Complete blood test

Annually:

• Complete baseline examinations (as at the beginning)
• Comprehensive imaging

Function - medically and technically explained

1st Hippo-Pathway

The normal function of the Hippo-Pathway is as follows:

Hippo signaling pathway active
    ↓
YAP1/TAZ are phosphorylated and inactivated
    ↓
Transcription of growth genes is stopped
    ↓
Apoptosis (cellular suicide) or cell cycle arrest
    ↓
Tumor does not grow

In cancer (disturbed Hippo pathway):

Procedure in the event of a disrupted Hippo pathway, e.g. cancer:

Hippo signaling pathway inhibited/mutated
    ↓
YAP1/TAZ remain active (dephosphorylated)
    ↓
Uncontrolled transcription of growth genes
    ↓
Cellular growth is hyperactive
    ↓
Cancer grows uncontrolled

Taking Huaier activates the polysaccharides and metabolites of Huaier:

• LATS1/2 kinases (Upstream regulators of the Hippo pathway)
• This re-phosphorylated YAP1/TAZ
• YAP1/TAZ become again inactivated
• The normal cell cycle control mechanism is restored

2. correction of transcriptional dysregulation

Thousands of genes are switched off in cancer: Genes that should be switched on are switched off and vice versa.
Huaier reactivates the transcription factors:

• NF-κB (Controls immune response and cell survival)
• c-Myc, Oct3/4, Sox2, Klf4 (Pluripotency factors - activate stem cell function)
• p53 (tumor suppressor - induces apoptosis)
• TCF/LEF (Wnt signal pathway effectors)

In addition, mass gene expression is reversed (within 4 weeks according to the Tanaka study)

12,000 to 25,000 new genes (normal cells only have ~20,000 in total) and 8,000 to 15,000 are silenced (switched off)

This leads to a massive „reprogramming“ of the cancer cell:

• Return to stem cell-like properties (non-differentiated)
• Apoptosis pathways are activated
  Or:
• Differentiation into the normal cell type (cell specialization) takes place

Through the reactivation of stem cell genes (c-myc, Oct3/4), the cancer cell becomes sensitive to normal control mechanisms again.

3. PI3K/AKT/mTOR signal path modulation

Normal (inhibited):

PI3K active → AKT active → mTOR active → Cell growth inhibited ✓
(This is oversimplified, but the concept)

For cancer (hyperactive):

PI3K overactive → AKT overactive → mTOR hyperactive → Uncontrolled growth ✗
(This is one of the most common defects in cancer cells)

Huaier has the effect that

• PTEN activated (negative regulator of PI3K)
• TSC1/TSC2 complexes become restored (inhibit mTOR)
• PI3K/AKT/mTOR is converted into normal Balance returned
• Cell growth becomes controllable again

Note: This route is particularly suitable for HER2-negative and Triple negatives Breast cancer excessively active.

---

4. miRNA- and piRNA-mediated transcriptional control

MicroRNAs (miRNA, small pieces of RNA (molecules) with a length of 20-22 nucleotides) are normally the „brakes“ for faulty genes. In cancer, these brakes are disrupted:

• Oncogenes are no longer inhibited
• Tumor suppressor genes are slowed down too much

Huaier provides the Restoration of miRNA function:

• miR-122 (inhibits HCC growth)
• miR-145 (inhibits stem cell genes in normal cells)
• miR-17/92 cluster (is activated by c-myc, can then induce apoptosis)

New miRNAs are activated, which:

• Oncogenes (e.g. KRAS, PIK3CA) shut down
• Tumor suppressor genes (TP53, RB) reinforce
• Angiogenesis (blood vessel formation) inhibit
• Epithelial-Mesenchymal Transition (EMT) block → Blocks metastasis

Tanaka study: Hundreds of new miRNA-variants that specifically „mute“ cancer cells.

---

5. immune activation (innate immune system)

β-Glucans as pattern recognition ligands:

β-Glucans (from Huaier)
    ↓
Binding to Dectin-1 and TLR receptors on immune cells
    ↓
Activation of macrophages and NK cells
    ↓
Secretion of pro-inflammatory cytokines:
    - TNF-α (tumor necrosis factor)
    - IL-12 (interleukin-12)
    - IFN-γ (interferon-gamma)
    ↓
Activation of cytotoxic T cells (CD8+)
    ↓
Recognition and lysis of tumor cells

The immune system is virtually „shaken awake“ and recognizes cancer cells as enemies again.

---

6. blockade of epithelial-mesenchymal transition (EMT)

The EMT process causes cancer cells to lose adhesion to their base, allowing them to migrate around the organism and lead to metastasis:

• Cells lose E-Cadherin (cellular adhesive)
• Cells express vimentin (movement protein)

Huaier provides the

• Stabilization from E-Cadherin (cells „stick“ together again)
• curtailment from Vimentin (cells can „migrate“ less)
• inhibition from Snail-, Slug- and Twist factors (EMT inducers)
• Stabilization from β-Catenin (maintains normal epithelial function)

This mechanically blocks the formation of metastases, even in the case of existing lymph node metastases.

7. blockage of angiogenesis (blood vessel formation)

Tumors can only grow if they form new blood vessels (angiogenesis). This is driven by VEGF (vascular endothelial growth factor).

Huaier counteracts this by

• VEGF expression inhibited becomes
• VEGFR signaling pathways blocked become
• HIF-1α (hypoxia inducible factor) downregulated becomes
• alternative pro-angiogenic pathways (FGF, Notch) inhibited become

Result: The tumor loses its blood supply - tumor growth is inhibited.

8. ribosomal RNA structure repair

The problem after chemotherapy:

• Chemotherapeutic agents, especially platinum complexes such as cisplatin, destroy ribosomal RNA structures
• Ribosomes are the protein factories of the cell
• Without functional ribosomes, the cell cannot produce proteins

Even if the tumor dies, healthy cells cannot regenerate

Huaier intervenes here and

• repairs ribosomal RNA structures
• provides the Restore protein synthesis capacity

This allows healthy cells to regenerate, while cancer cells die again. This explains why Huaier patients undergoing chemotherapy have fewer side effects and recover more quickly.

Immunologically relevant genes

Regulatory behavior of genes

Genes can be linked to 0% Expression (corresponds practically to OFF) or with any percentage of their maximum capacity, or with 100% Expression (for complete AN).

Tumor necrosis factor α

The regulatory behavior and its consequences are explained using the example of the tumor necrosis factor α (TNFα):

The normal “value“ is 40% expression, enough to protect against infections, too little to attack tissue.

If the value rises to 100% (or even higher), e.g. in rheumatoid arthritis, this results in

• a massive excess of TNF-α
• Permanent joint inflammation
• Destruction of cartilage and bone
• systemic inflammation

and the symptoms of permanent joint pain and swelling.

If, however, the value is reduced to only 5%, for example, then this results in

• Too little TNF-α to kill pathogens
• Unlimited bacterial growth
• Systemic organ failure
• Fatalities possible

Conclusion: TNF is vital!

Spectrum of the cytokine IL-6 (interleukin 6)

• Silent – 0-5% des Kontrollwertes
  No acute phase reaction, no fever
  Infection blindness
• Very quiet – 5-15% des Kontrollwertes
  Minimal inflammatory response
  Weak immunity
• Quiet – 15-30% des Kontrollwertes
  Mild local inflammation
  NORMAL after a minor infection
• Moderate – 30-50% des Kontrollwertes
  Clear but limited inflammation
  NORMAL in case of infection
• Loud – 50-80% des Kontrollwertes
  Severe systemic inflammation
  Too much? For RA, IBD
• Very loud – 80-95% des Kontrollwertes
  Massive systemic inflammation
  Sepsis, shock
• Maximum – 95-100%+ des Kontrollwertes
  Cytokine storm, organ failure
  Fatal (COVID-19, sepsis)

Beispiel für zu niedrige Expression

• TNF-α bei 90% statt 40% des Kontrollwertes
  Autoimmun-Entzündung
• IL-17 bei 85% statt 30% des Kontrollwertes
  Überproduktion von Th17 führt zu überschießenden Entzündungsreaktionen
• IL-6 bei 95% statt 45% des Kontrollwertes
  Chronische Arthritis

Beispiel für zu hohe Expression

• TNF-α bei 10% statt 40% des Kontrollwertes
  Tuberkulose-Risiko
• IL-10 bei 8% statt 35% des Kontrollwertes
  Entzündung unkontrolliert
• IFN-γ bei 12% statt 50% des Kontrollwertes
  Virale Anfälligkeit

Messmethoden

Die Regulation von Genen erfolgt auf mehreren biologischen Ebenen. Um diese Ebenen zu verstehen, gibt es vier Hauptmessmethoden, die verschiedene Aspekte der Gen-Expression quantifizieren:

1. Ebene 1: Transkription (DNA → mRNA)
   Messmethode: qRT-PCR
   
2. Ebene Proteinproduktion (mRNA → Protein in der Zelle)
   Messmethode: Western Blotting
   
3. Ebene 3: Sekretion/Zirkulation (Protein im Serum/Plasma)
   Messmethode: ELISA
   
4. Ebene 4: Zelluläre Expression auf Einzelzell-Ebene
   Messmethode: Flow Cytometry

1. qRT-PCR (Quantitative Reverse Transcription PCR)

qRT-PCR misst die Menge der mRNA in Zellen oder Geweben in der Messgröße „Vielfaches“

• Fold-Change (Vielfaches): Beispiel: TNF-α mRNA ist 2.5-fold erhöht
  • Bedeutung: 2.5× höher als die Kontroll-Gruppe
  • Ein Wert von 0.45 bedeutet: 45% der Kontrolle (also herunterreguliert)
• Cycle Threshold (Ct): Rohwert, wie viele PCR-Zyklen bis zur Detektion nötig sind
  • Niedrigere Ct = mehr mRNA vorhanden
  • Höhere Ct = weniger mRNA vorhanden

Was qRT-PCR NICHT misst:

• die absolute Menge des Proteins
• die Aktivität des Proteins
• ob das Protein sekretiert wurde
• die Konzentration im Serum

Klinische Interpretation

qRT-PCR: TNF-α = 2.5-fold

Bedeutet: "TNF-α mRNA ist 2.5× höher als normal"
          "Der Gen-'Lautstärkeregler' ist lauter gestellt"
          
ABER: Das sagt NICHTS über die tatsächliche TNF-α-Proteinmenge im Serum aus!
Flow Cytometry zeigt, dass auch bei hoher mRNA nicht automatisch viel Protein pro Zelle entsteht, und selbst wenn, muss es noch sekretiert werden. Die 8.3-fold mRNA-Erhöhung im folgenden Beispiel kann also zu viel oder wenig Protein in den Zellen führen.

Praktisches Beispiel

Patient mit bakterieller Infektion:
qRT-PCR (Blut-Leukozyten): TNF-α = 8.3-fold erhöht
→ Die Zellen produzieren viel mRNA
→ die aber nicht sofort im Serum messbar ist
→ denn das Protein kommt erst nach etwa 30 Minuten bis Stunden im Serum an.

2. Western Blot

Der Western Blot misst die Menge von Protein innerhalb von Zellen oder Geweben in der Messgrlße „Bandintensität“, den Phosphorylierungsstatus (aktiviertes vs. inaktives Protein) und verschiedene Protein-Isoformen.

• Relative Bandintensität: 0-100% oder als Vielfaches zur Kontrolle
• Beispiel: IL-6 Protein = 65% der Kontroll-Intensität
  • Bedeutung: Das Protein ist zu 65% so stark exprimiert wie in der Kontrolle

Was Western Blotting NICHT misst:

• ob das Protein aktiv ist (nur Präsenz)
• ob das Protein sekretiert wurde
• die Konzentration im Serum/Blut
• auf Einzelzell-Ebene

Klinische Interpretation:

Western Blot: TNF-α Protein = 72% der Kontroll-Intensität

Bedeutet: "TNF-α Protein ist zu 72% im Zell-Lysat vorhanden"
          "72% so viel Protein wie in der Kontroll-Zellkultur"
          
ABER: Das sagt NICHTS über:
      - Wie viel TNF-α tatsächlich sekretiert wurde
      - Wie viel TNF-α im Serum ist
      - Ob das Protein aktiv ist oder nicht

Praktisches Beispiel:

Makrophagen-Kultur mit LPS-Stimulation:
Western Blotting (Zelllysat): TNF-α = 85% der Kontrolle
ELISA (Kulturüberstand): TNF-α = 2,800 pg/mL

Fazit: Es wurde viel TNF-α Protein hergestellt UND sekretiert

3. ELISA (Enzyme-Linked Immunosorbent Assay)

ELISA misst die absolute Konzentration von Protein im Serum, Plasma, Zellkultur-Überstand oder anderen Körperflüssigkeiten in absoluter Konzentration.

• pg/mL (Pikogramm pro Milliliter)
  für Zytokine wie TNF-α, IL-6
• ng/mL (Nanogramm pro Milliliter)
  für konzentriertere Proteine
• µg/mL (Mikrogramm pro Milliliter)
  für sehr hohe Konzentrationen

Normalwerte für TNF-α (Beispiel):

Gesund:              < 5-20 pg/mL
Leichte Infektion:   20-100 pg/mL
Moderate Infektion:  100-500 pg/mL
Schwere Infektion:   500-5,000 pg/mL
Sepsis/Cytokine-Storm: > 5,000 pg/mL (kann tödlich sein)

Was ELISA misst:

• absolute Menge des sekretierten/zirkulierenden Proteins
• ob das Protein tatsächlich im Blut/Serum angekommen ist
• die systemische Auswirkung (nicht nur lokal in der Zelle)

Was ELISA NICHT misst:

• wie viel mRNA vorhanden ist
• wie viel Protein in den Zellen ist
• ob das Protein aktiv ist
• auf Einzelzell-Ebene

Klinische Interpretation:

ELISA: TNF-α im Serum = 65 pg/mL

Bedeutet: "Es sind 65 Pikogramm TNF-α pro Milliliter Serum vorhanden"
          "Das ist 3-13× über dem Normalwert"
          "Es liegt moderate Entzündung vor"
          
Dies ist eine ABSOLUTE Konzentration, nicht relativ!

Praktisches Beispiel:

Patient mit rheumatoider Arthritis:
ELISA: TNF-α = 85 pg/mL (normal: < 20 pg/mL)
qRT-PCR (Blut): TNF-α mRNA = 3.2-fold erhöht
Western Blotting (Gelenksynovia): TNF-α = 95% (sehr hoch lokal)

Fazit: Überall zu viel TNF-α,von der mRNA über zelluläres Protein bis zum Serum

4. Flow Cytometry

Flow Cytometry misst die Expression von Proteinen oder Markern auf der Oberfläche oder im Inneren einzelner Zellen in Prozent und Fluoreszent-Intensität.

• % positive Zellen: Beispiel: 78% von CD4+ T-Zellen exprimieren IL-2
  • Bedeutung: 78% dieser Zellpopulation hat das Merkmal
• Mean Fluorescence Intensity (MFI): 0-10,000+ (je nach Instrument)
  • Beispiel: IL-2 Expression MFI = 450 in CD4+ T-Zellen
  • Höhere MFI = mehr Protein pro Zelle

Was Flow Cytometry misst:

• Wie viele Zellen einer bestimmten Population ein Antigen exprimieren (%)
• Wie viel Antigen pro Zelle vorhanden ist (MFI)
• Zelluläre Heterogenität (nicht alle Zellen sind gleich!)
• Zelloberflächen-Marker und intrazelluläre Proteine

Was Flow Cytometry NICHT misst:

• Die Serum-Konzentration (misst Zellen, nicht Serum)
• Die mRNA-Menge – Wie viel insgesamt im Körper insgesamt ist –
  Mit zusätzlichen Daten (Zellzahl, Gewicht, etc.) kann man indirekt hochrechnen:
  Diese Hochrechnung ist jedoch nur eine Schätzung, nicht so exakt wie ELISA
  und sie erfasst nur die gemessenen Zellen (z.B. Blut-Makrophagen), nicht Gewebe-Makrophagen!

Klinische Interpretation:

Flow Cytometry: 73% von CD8+ T-Zellen exprimieren IFN-γ
                MFI = 520

Bedeutet: "73% der cytotoxischen T-Zellen haben IFN-γ Protein"
          "Der durchschnittliche IFN-γ-Gehalt pro Zelle ist 520 (MFI)"
          "Die T-Zell-Antwort ist aktiv"
          
ABER: Das sagt NICHTS über:
      - Wie viel IFN-γ insgesamt im Serum ist
      - Wie viel IFN-γ mRNA vorhanden ist

Praktisches Beispiel:

COVID-19 Patient (Tag 3 nach Infektion):
Flow Cytometry: 
  - 91% CD8+ T-Zellen exprimieren IFN-γ (high!)
  - MFI = 1,250 (sehr hoch)
  
ELISA: IFN-γ im Serum = 180 pg/mL (normal: < 50)

Fazit: Starke T-Zell-aktivierte IFN-γ-Produktion, systemisch messbar