CN117562937A

CN117562937A - Application of defatted flaxseed powder in the preparation of drugs against new coronavirus infectious diseases

Info

Publication number: CN117562937A
Application number: CN202311495813.8A
Authority: CN
Inventors: 刘树林; 赵营; 苑晓慧; 单万亭; 张幸华
Original assignee: Harbin Tianqi Human Second Genome Technology Development Application Technology Co ltd
Current assignee: Harbin Tianqi Human Second Genome Technology Development Application Technology Co ltd
Priority date: 2023-11-10
Filing date: 2023-11-10
Publication date: 2024-02-20

Abstract

The invention relates to the application of defatted flaxseed powder in the preparation of anti-coronavirus infectious disease drugs, and belongs to the field of biomedicine technology. In order to solve the problem of adverse reactions in existing drugs for treating new coronavirus infectious diseases, the present invention provides the application of defatted flaxseed powder in the preparation of anti-new coronavirus infectious disease drugs. The defatted flaxseed in the anti-new coronavirus infectious disease drugs The content of powder is 0.01~99.9wt.%. The present invention uses defatted flaxseed powder to treat novel coronavirus infectious diseases. The defatted flaxseed powder can be converted into enterodiol and enterolactone under the action of intestinal microorganisms. The antiviral effect of enterodiol is significantly better than that of chemical drugs, because it mobilizes the body's own immunity and is non-antagonistic antiviral, so it has no toxic side effects, is low-cost, easy to take, and has significant therapeutic and preventive effects. It is expected to achieve multi-year human The beautiful vision of anti-virus without harming body cells is worthy of clinical promotion.

Description

Application of defatted flaxseed powder in preparing medicine for treating new coronavirus infectious diseases

Technical Field

The invention belongs to the technical field of biological medicines, and particularly relates to application of defatted flaxseed powder in preparation of medicines for resisting new coronavirus infectious diseases.

Background

Some antiviral drugs have been used for the treatment of novel coronavirus infectious diseases such as interferon, oseltamivir, lopinavir, ritonavir, fepinat Weili bavin, adefovir, chloroquine, arbidol, etc.; however, most of the existing medicines have adverse reactions with different degrees, and especially for asymptomatic infected persons or mild patients, the clinical benefit risk ratio is low. In addition, the detection of the patient's novel coronavirus nucleic acid occurs when the patient turns negative and then returns positive; after part of the patient respiratory tract specimen virus nucleic acid turns negative, the fecal specimen is still positive for a long time and is reported; the present antiviral drugs are not capable of well controlling digestive system infection of the new coronaviruses. Persistent infection with new coronaviruses is an important factor in impeding patient recovery. Rapid removal of residual virus in infected individuals is critical to achieving a negative turn for new coronavirus nucleic acid detection.

How to provide a drug which is basically harmless to the organism and can treat the infectious diseases of the new coronaviruses is a problem to be solved in the field at present.

Disclosure of Invention

In order to solve the problem of adverse reaction of the existing novel coronavirus infectious disease treatment drugs, the invention provides application of defatted flaxseed powder in preparation of drugs for resisting the novel coronavirus infectious disease.

The technical scheme of the invention is as follows:

application of defatted Linseed powder in preparing medicine for treating infectious diseases caused by new coronavirus is provided.

Further, the defatted flaxseed powder is obtained by crushing defatted flaxseed meal to 80-120 meshes after flaxseed oil extraction by a low-temperature cold pressing method.

Further, the content of defatted flaxseed powder in the drug for resisting new coronavirus infectious diseases is 0.01-99.9 wt%.

Further, the content of the effective medicine component of defatted flaxseed in the medicine for treating the infectious diseases of the new coronavirus is 0.01 to 99.9 wt%.

Further, the extraction method of the effective medicine components of the defatted flaxseed comprises the following steps: adding the defatted flaxseed powder into pure water according to the mass ratio of the defatted flaxseed powder to the pure water of 1:5-10, heating to 100 ℃ and boiling for 2min to obtain water extract or drying the obtained water extract to obtain the water extract, namely the effective pharmaceutical ingredient of the defatted flaxseed.

Furthermore, the medicine for resisting the new coronavirus infectious diseases also contains a pharmaceutically acceptable carrier or excipient.

Further, the pharmaceutically acceptable carrier includes a liposome, micelle, dendrimer, microsphere, or microcapsule.

Further, the preparation form of the drug for resisting the new coronavirus infectious diseases is tablets, granules, capsules, solutions, pills, liposome preparations or nanoparticle preparations.

Furthermore, the medicine for resisting the new coronavirus infectious diseases is administrated in an oral mode, wherein the oral dosage of the medicine is 40g calculated by defatted flaxseed powder, and the oral dosage is 1-2 times per day.

The invention has the beneficial effects that:

the defatted flaxseed powder is used for treating new coronavirus infectious diseases, and lignans contained in the defatted flaxseed powder can be converted into enterodiol END and enterolactone ENL under the action of intestinal microorganisms. The enterodiol antiviral effect is obviously superior to that of chemical medicines, and is non-antagonistic antiviral by regulating the autoimmune power of human body, so that the enterodiol antiviral effect has no toxic or side effect, and is expected to realize the expected antiviral effect of human for many years and have good prospect of no harm to body cells. Intestinal glycols are not currently used for pharmaceutical purposes worldwide due to their high price. The method for treating the novel coronavirus infectious diseases by converting the defatted flaxseed powder into the enterodiol has the characteristics of low cost, convenient administration and remarkable treatment and prevention effects, and is worthy of clinical popularization.

Drawings

FIG. 1 is a photograph of HE staining of large intestine tissue of a lignan-fed mouse;

FIG. 2 is a photograph of HE staining of normal mouse large intestine tissue;

FIG. 3 is a graph showing comparison of the abundance of END in the intestinal tract of example 1;

FIG. 4 is a graph showing comparison of the abundance of ENL in intestinal tract in example 1.

Detailed Description

The following embodiments are used for further illustrating the technical scheme of the present invention, but not limited thereto, and all modifications and equivalents of the technical scheme of the present invention are included in the scope of the present invention without departing from the spirit and scope of the technical scheme of the present invention. The process equipment or apparatus not specifically noted in the following examples are all conventional equipment or apparatus in the art, and the raw materials and the like used in the examples of the present invention are commercially available unless otherwise specified; unless specifically indicated, the technical means used in the embodiments of the present invention are conventional means well known to those skilled in the art.

Example 1

This example examined the clinical efficacy of defatted flaxseed meal in treating patients positive for new coronavirus infection.

The defatted linseed powder used in this example was obtained from red light oil plants in Shanxi county, heilongjiang province, and was pressed twice with a cold rolling mill to remove 70% of oil. Obtaining flaxseed cake, and grinding into 80-120 mesh fine powder by a grinder. Meets the quality standard of defatted flaxseed powder.

The experiments were divided into three groups:

model group: patients with a new coronavirus infection (nucleic acid positive) were selected while 40g of ordinary protein powder was administered orally per day.

SECO group: patients with a new coronavirus infection (nucleic acid positive) were selected while 40g of defatted flaxseed meal was orally administered daily.

Control group: normal persons not infected with the new coronavirus (nucleic acid negative) were selected while 40g of ordinary protein powder was orally administered per day.

Patients in SECO group take plant lignans, namely defatted flaxseed powder, 40g of defatted flaxseed powder is added into 10 times mass of pure water, heated to 100 ℃, boiled for 2 minutes, taken after cooling, 1-2 times a day for 7 days.

After continuously taking defatted flaxseed powder for one week, collecting 30g of feces of three groups before taking and on day 7 of taking, freezing at-18deg.C, and detecting intestinal diol content in feces by genome center of Harbin university of medical science.

Non-targeted metabolite detection (n=5) was performed on feces using ultra-high performance liquid chromatography tandem fourier transform mass spectrometry (LC-MS/MS), and MS and MSMS mass spectrometry information was matched to the metabolic public databases HMDB (http:// www.hmdb.ca /) and Metlin (https:// Metlin. Scripps. Edu /), resulting in metabolite information, and their relative abundance was calculated.

It was found by non-targeted metabolite detection that administration of defatted flaxseed meal 40g per day orally in patients with a new coronavirus infection (nucleic acid positive) significantly increased the relative abundance of END, ENL in the patient's feces compared to Model group (p < 0.001).

The relative abundance of END and END in the feces of each group of patients is statistically analyzed (single-factor analysis of variance), and after the defatted linseed powder is taken by the patients of the treatment group, the relative abundance of END and END in the feces of the SECO group is obviously improved compared with the Model group, so that the effect of bacteria in intestinal tracts of the defatted linseed powder can generate END and END, and the relative abundance of END and END in the intestinal tracts is further improved to achieve the treatment purpose.

Meanwhile, the clinical symptoms and assay detection indexes of the patient with positive coronavirus infection are observed and recorded, and the clinical symptoms of the patient are obviously improved until the patient is cured after taking the medicine for one week.

Typical cases 9 cases:

case 1, wang Mou, male, 88 years old. Identification number 23010719351110XXXX. The 2023, 6 and 2 days check the positive of nucleic acid. Symptoms: fever was 37.8 degrees, general weakness, slight cough, chest CT indicated that both lungs were scattered in flaky shadows, and a small amount of effusion in the right chest. 40g of defatted flaxseed powder is taken, decocted in water for 2 minutes, cooled, taken once a day, after taking for two days, fever is resolved, the whole body is hypodynamia is relieved, the sleeping is good, cough is relieved, the CT shadow of the lung is relieved after one week, and other antibiotics are not used.

Case 2, xu Mouhua, female, 88 years old. Identification number 23010719350418XXXX. The 2023, 6 and 2 days check the positive of nucleic acid. Symptoms: fever is 38.8 degrees, general weakness, pain, cough, chest CT shows changes in double lung interstitial inflammation. 40g of defatted flaxseed powder is taken, decocted in water for 2 minutes, cooled, taken once a day, after taking for two days, fever is resolved, the pain of the whole body is relieved after taking for three days, the sleeping is good, the cough is relieved, the CT interstitial inflammation of the lung is relieved after one week, and other antibiotics are not used.

Case 3, li Mouju, female, 53 years old. Identification number 23010719690526XXXX. Fever began on day 5 and 15 of 2023, body temperature 38.6 degrees, and positive nucleic acid was checked. General weakness and pain, cough, poor appetite, diarrhea, and pharyngalgia. No abnormalities were seen in chest CT. 40g of defatted flaxseed powder is taken, decocted in water for 2 minutes, cooled, taken once a day, after taking for two days, fever is resolved, and after taking for three days, the whole body is hypodynamia, pain is relieved, sleeping is good, cough is relieved, pharyngalgia is relieved, and diarrhea is stopped. No other medication was used.

Case 4, zhang Fu J, female, 53 years old, identification 23050519691103XXXX. Fever 38 degrees in 2022, 12 and 13 days, cough, pharyngalgia, fatigue and weakness, palpitation and positive nucleic acid detection. The defatted flaxseed powder is taken with 40g of water for decoction every day. The following day, debilitation is relieved, the third day starts to abate fever, the fever gradually improves, and the symptoms completely disappear until the sixth day.

Case 5, wu Shu F, female, 60 years old, identification 23232419621129XXXX. Fever 38.2 degrees in 2022, 12 and 25 days, cough, pharyngalgia, general fatigue and weakness, palpitation and positive nucleic acid detection. The defatted flaxseed powder is taken with 40g of water for decoction every day. The next day, debilitation is relieved, and the third day begins to abate fever, gradually improves, and symptoms completely disappear.

Case 6, sun Chengxian, men, 78 years old, identification 23010319440731XXXX. Fever 37.8 degrees in 2022, 12 months, 24 days, cough, pharyngalgia, poor appetite, general fatigue and weakness, palpitation and positive nucleic acid detection. The defatted flaxseed powder is taken with 40g of water for decoction every day. The next day, the debilitation is relieved, the sleep is good, the stool is smooth, the fever starts to abate in the third day, the fever gradually improves, and the symptoms completely disappear.

Case 7, zhao Fenghua, female, 54 years old, identification 23213119681021XXXX.2022, 12, 24 days fever 39.4 degrees, cough, sore throat, poor appetite, general fatigue and weakness, palpitation, and positive nucleic acid detection. The defatted flaxseed powder is taken with 40g of water for decoction every day. The next day, the debilitation is relieved, the sleeping is better than that of the weekdays, the fever is reduced, the fever is gradually improved, and the symptoms are completely disappeared.

Case 8, li Huaxue, men, 77 years old, identification 23232519451123XXXX.

Onset of disease at 26/12/2022, body temperature of 37.6, pharyngalgia, cough, palpitation, anorexia, and general soreness and debilitation. The defatted flaxseed powder is taken with 40g of water for decoction every day. The sleep is better than that of the weekdays, the hypodynamia is relieved, the fever is reduced, the cough is relieved, the appetite is increased, and the effect is improved.

Case 9, liu Jun, men, 51 years old, identification 23210319710504XXXX.

Onset of disease at 28 days of 12 months 2022, body temperature 38.6, pharyngalgia, cough, palpitation, anorexia, and general soreness and debilitation. The defatted flaxseed powder is taken with 40g of water for decoction every day. The third day starts to abate fever, the appetite is increased, the sleep is better than that of the weekdays, the hypodynamia is relieved, the fever is obviously improved from the fifth day to the seventh day, and the symptoms are completely eliminated.

Control group, 7 cases of illness state without defatted linseed powder were returned:

case-1, fangbing, female, 30 years old. An identity card: 23070319931003XXXX.

2022, 12, 25 and fever 38.0 degrees. Sore throat, cough, palpitation, weakness, anorexia, and general soreness. The temperature of the next day is reduced to 37.1 ℃ after 1 granule of ibuprofen is taken. Pain began to abate after 8 days. Cough, debilitation and general soreness is relieved after 14 days.

Case-2, liu Xianglan, female, 59 years old. An identity card: 23100219631227XXXX. Nucleic acid positive at 2022, 11 and 3. Body temperature 38.2 degree, pharyngalgia, cough with little sputum, poor appetite, debilitation and muscular soreness. The oral tablet for relieving pain has body temperature reduced to 37.6 deg.C. Only 2000 ml of water is drunk every day, and no other medicines are used. Symptoms gradually abate 15 days after fever.

Case-3, xue Huina, female, 40 years old. Identification card 23070219820318XXXX. Nucleic acid positive at 2022, 11 and 3 days with fever of 38.9 degrees. Cough with white phlegm, sore throat, loss of appetite, weakness of the whole body and soreness of muscles. The body temperature is reduced to 37.4 ℃ when one ibuprofen granule is taken, and the cefalexin capsule is taken for 2 granules 3 times a day. Slightly improved appetite by day 8 and reduced after 10 coughs. The muscle soreness is relieved to cure after 18 days.

Case-4, ma Hailong, men, 50 years old, identification 23012119720227XXXX. Nucleic acid positive at 11 and 17 days 2022, fever 39 degrees, cough, sore throat, poor appetite, nausea, and general debilitation and aching pain. 1 granule of ibuprofen is taken 2 times a day, and the body temperature is reduced to 37.4 ℃. The effect is poor because 0.5 g of levofloxacin is added once a day for 5 consecutive days. After 12 days, the symptoms gradually lessened.

Case-5, zhao Jing, female, 34 years old. Identification card 23230119880923XXXX.

Nucleic acid positive at 2022, 11 and 11. Fever 39.2 degrees, sore throat, cough, chest pain, nausea, general debilitation and aching pain, poor appetite. 1 granule of ibuprofen is taken twice a day for 2 consecutive days, and the body temperature is reduced to 37.6 ℃. The antibiotic is orally taken in 2 tablets of Baifule three times a day. Cough is relieved after seven days, and appetite is improved. The sore throat, the debilitation and the ache of the whole body are gradually improved 14 days after the illness.

Case-6, yao Min, female, 39 years old. Identification card 23010719830323XXXX. Nucleic acid positive at 2022, 11 and 13. Fever of 38.9 degrees, sore throat, cough, nausea, poor appetite, debilitation and aching pain of the whole body. Ibuprofen 1 granule is taken twice a day, and the body temperature is reduced to 37.6 ℃. After 8 days, the cough is relieved, and the appetite is improved. The sore throat, the debilitation and the ache of the whole body are gradually improved 14 days after the illness.

Case-7, lv Baoshan, male, 60 years old. Identification card 2301071919620324XXXX. Nucleic acid positive at 2022, 12 and 13. Fever of 38.8 degrees, sore throat, cough, nausea, poor appetite, debilitation and aching pain of the whole body. The pain relieving tablet 1 tablet is taken twice a day, and the body temperature is reduced to 37.5 ℃. After 9 days, the cough is relieved, and the appetite is improved. The sore throat, the debilitation and the ache of the whole body are gradually improved 15 days after the illness.

The above cases were delegated to the Heilongjiang kang Yuan neurologic specialty hospital for treatment and recording.

The intestinal flora can metabolize the secoisolariciresinol diglucoside (phytolignans) in defatted flaxseed powder into the mammalian lignans (enterodiol END) through biotransformation, and the main steps comprise hydrolysis and deglycosylation to produce ring-opened secoisolariciresinol, and then demethylation and dehydroxylation to produce the mammalian lignans enterodiol. After eating for 12 hours, the conversion products appear, and after 18-24 hours, the conversion peak is reached.

In vivo transformation is based on the symbiotic relationship of the intestinal flora with the human host, both of which benefit. Specific biochemical mechanisms include multiple metabolic steps of deglycosylation, demethylation, dehydroxylation, etc., with the end result that humans provide habitat (gut) and food (defatted flaxseed, etc.) for these bacteria, which provide beneficial metabolites (animal lignans such as enterodiol and enterolactone, etc.) for human hosts. In vivo transformation was complete and thorough, transformation was 100% successful.

Furthermore, the inventor of the application researches that the flora participating in bioconversion is not high in abundance before the defatted flaxseed is eaten, so that the flora is in a waste state. Once defatted flaxseed is ingested, the flora involved in bioconversion will become active and the abundance will increase dramatically. Thus, the amount of the transforming flora involved in the body is dynamic, depending on the ingestion of defatted flaxseed, the final amount of intestinal microbiota is sufficient to ensure complete transformation of the ingested defatted flaxseed. The use of defatted flaxseeds for bioconversion is not conventional with pure flaxseeds because the fat in the pure flaxseeds before degreasing is a disturbing factor and costs up to a hundred times, and more importantly, the endless fat intake is detrimental to health.

Example 2

In the embodiment, the mouse experiment proves that the taking of the defatted flaxseed powder can improve the immunity of the organism, thereby improving the capability of the organism to resist the infection of the new coronavirus.

By feeding the defatted flaxseed powder to model mice, the antiviral mechanism of lignans contained in the defatted flaxseed powder was found to have the following two points:

(1) Lignans can regulate body immunity and promote immune infiltration reaction;

(2) By modulating the intestinal flora, the intestinal immune response is activated.

This example presents a "relative dose of infection" experiment to demonstrate that defatted flaxseed meal contains lignans that enhance body immunity.

The basic principle of this embodiment is as follows:

there is a rough range of infectious doses for any microbial pathogen to infect a particular host, which is the "natural infectious dose (Natural infectious dose)" developed during evolution. "relative infection dose (Relative infectious dose)" means that after the host's resistance has been enhanced, the original infection dose has not been able to cause infection in the host, and more than 1000 times the original infection dose is required to cause infection; this "new" infection dose is due to the fact that the host's resistance is improved by more than 1000 fold over the original state. The measure of improving host resistance of the team is to use lignans to condition intestinal flora so as to achieve the optimal functional state and cooperate with an immune system to resist attack of pathogenic microorganisms.

The test protocol of this example is as follows:

the experiment uses mice as experimental animals and Salmonella typhimurium (Salmonella paratyphi C RKS 4594) as pathogens to infect the mice.

Design of experiment

First, half of the infection amount of the salmonella paratyphi RKS4594 strain of the C-type Salmonella against the experimental animals was determined by a preliminary experiment as "natural infection dose". Based on this, an experimental group was designed.

Experimental animals: 160 Balb/C mice

Bacterial strain: salmonella paratyphi C RKS4594

Experimental grouping:

1. infection control group: infecting 20 animals with a "natural infective dose";

2. infection-free control group: phosphate buffer sham "infects" 20 animals;

3. experimental group: the bacterial inoculum size of the natural infection dosage, 10 times, hundred times, thousand times, ten thousand times and hundred thousand times of the natural infection dosage is 20 respectively.

Administration of lignans: animals in the experimental group received lignans and animals in the control group were perfused with phosphate buffer.

The experimental procedure is as follows:

1. lignans condition the intestinal flora:

animals of the experimental group received lignans (0.5 ml lavage, once daily) to condition the intestinal flora for 3 weeks, and animals of the control group and the non-infected control group were lavaged with 0.5 ml phosphate buffer instead of lignans as controls.

2. Preparing bacterial liquid:

1) Strains stored at-80℃were streaked in three sections on solid LB medium and incubated overnight at 37 ℃.

2) Single colonies were selected and cultured with three 3ml liquid LB medium to log phase overright (16-18 hours),

3) At this time, the concentration was 5X 10 ⁹ /ml (stock solution).

4) Taking 10 μl of stock solution, adding 2490 μl of fresh LB culture solution, mixing to obtain 2×10 ⁷ /ml。

5) The concentration of the mixture is 2 multiplied by 10 after being diluted by equal ratio ⁶ /ml、2×10 ⁵ /ml、2×10 ⁴ /ml。

3. Treatment of mice:

1) Mice were housed in separate cages and marked with markers.

2) Gastric lavage: each mouse was perfused 500 at the above concentrations. l (L)

3) Three days after infection, mice were sacrificed by cervical dislocation.

4. Grinding liver, spleen, kidney and lung

1) 160 groups of mortar sterilization treatments were prepared in advance.

2) PBS buffer, LB solid medium, 3ml LB liquid medium.

3) The mice were taken from the liver, spleen, kidney and lung, sheared, added with 1ml PBS buffer solution and ground thoroughly.

4) 100 μl of each stock solution and 10-fold serial dilutions of tissue solution were used to seal solid LB plates, and marked with 37 degrees of overlapping.

5) 100 μl of the remaining bacterial liquid was added to 3ml of the liquid medium for overnight culture. If no colony grows on the upper plate, the upper plate is then densely coated with liquid culture medium.

5. Peripheral blood test procedure

1. Blood collection

The animals were anticoagulated from the orbital vein Cong Caixie ml, EDTA-K2 under ether anesthesia after 12h of empty stomach, and were examined using a whole blood cell analyzer to obtain white blood cell and platelet counts. Taking liver, spleen, kidney, lung and blood bone marrow to culture bacteria; plate colonies were counted and liquid culture confirmed the presence or absence of bacteria.

2. Smear for painting

1. EDTA anticoagulated peripheral blood was pipetted in 5-7. Mu.l or mouse peripheral blood was collected directly and the blood was dropped to about 1CM at one end of the slide or 3/4 of the whole piece. The slide with frosted slice head is used, if the slide has grease, the slide is degreased by alcohol, and then is wiped or air-dried for standby. The pushing ends of the pushing sheets are flush.

2. The slide glass is held by the left hand and the slide glass is held by the right hand, the push piece is close to the blood drop, and then the blood drop is lightly contacted and pressed on the blood drop, so that the blood spreads in a U shape and fills the width of the push piece.

3. The pusher was angled 30 degrees from the slide and blood was pushed toward the other end of the slide at a uniform rate. When the hematocrit changes, the angle between the slide and the slide should be properly adjusted, as well as the speed at which blood is pushed. The standard blood smear should be made with distinct head, body and tail, with gaps left on both sides and ends.

3. After dyeing

1. The mixed staining of Rui-Giemsa combines the advantages of both Rui cytoplasmic staining and Giemsa nuclear staining.

2. The dyeing method comprises the following steps: marking a blood smear (HB pencil, frosted part), putting the blood smear on a staining rack horizontally, dripping A liquid to cover the whole blood membrane, dripping B liquid after 30 seconds-1 minute, uniformly mixing A, B liquid, lightly blowing a liquid level by using an ear washing ball, wherein the ratio of A, B liquid is 1:2, standing for 5-10 minutes at a horizontal position, taking up the glass slide, and gently shaking to ensure that the dye is not attached to the blood membrane any more. The water flow is regulated to the minimum, the glass slide is held flat, the water flow direction is 90 degrees, the glass slide is flushed in the head of the blood film, the angle of the glass slide is regulated towards the tail end of the blood film until the dye is flushed out, the flushed blood smear is placed on a blood slide frame to naturally dry the water, or the glass slide is inclined, and the glass slide is placed on two folded roll papers to absorb the water, and waits for natural drying.

3. Judging the dyeing result: under normal conditions, the good staining result is a bloodfilm appearance of light purple; under the low power lens, the cells are uniformly distributed; cells are pink, rarely stained particles, and white cell cells can display the characteristic colors of various cells, and white cell nuclei are stained into purplish red.

6 step of bone marrow detection

1. Bone marrow specimen collection

The mice are killed by cervical vertebra removal, two femur bone marrow of the mice are completely blown out, and femur with bone marrow is taken aseptically for bacterial culture.

2. Smear preparation

The extracted marrow tissue is rapidly smeared for 3 to 5 pieces, and the smeared blood film can be separated into a head part, a body part and a tail part for examination. The smear should be quickly fan dried after being made so as to avoid shrinkage and deformation of cells during natural drying. If histochemical staining is performed, the number of smears is increased.

3. Dyeing

Cell morphology is generally selected from Rayleigh staining and Giemsa staining (staining procedure is similar to blood platelets).

4. Judgment standard

The bone marrow smear has bone marrow granules, and special cells in the bone marrow such as megakaryocyte, plasma cell, osteoblast, osteoclast and the like can be seen under the microscope, and the ratio of the neutrophil-shaped granulocyte to the foliate granulocyte is larger than that of the blood smear, so that the above points indicate that the material is good.

5. Microscopic morphological examination

(1) Low power microscopy: (1) observing the conditions of sampling, smearing and staining. (2) The degree of myeloproliferation is determined based on the ratio of mature red blood cells to nucleated cells. (3) Observing whether special abnormal cells exist in the picture. (4) The whole megakaryocyte count was counted with a low power microscope.

(2) And (5) oil microscopic examination: (1) the presence or absence of abnormal cells and parasites was observed. (2) A satisfactory blood membrane site was selected, 200 or 500 nucleated cells were carefully observed, counted by cell type, developmental stage classification, and the percentage of each segment was calculated. (3) And calculating the particle-red ratio. (4) And checking whether the cells have pathological morphological changes at each system stage.

Experimental results

The pre-experiment result shows that the half-number infection rate of the mice in the batch is 10 by Salmonella paratyphi C RKS4594 ³ Bacteria/mouse. Based on this result, 10 was used ³ Individual bacteria/mice were vaccinated with lignan-free groups and with lignan-infused 3 week experimental groups. Each group had 20 mice. Table 1 shows the number of infected mice among 20 mice.

TABLE 1

Note that: 20 control mice were not vaccinated with bacteria, did not receive lignan lavage, and did not have infection.

Conclusion:

salmonella paratyphi C RKS4594 half-number of infected mice of this batch had an infection of 10 ³ Bacteria/mouse (natural infection dose). After conditioning the intestinal flora with lignans, the half-number of infections of Salmonella paratyphi C RKS4594 on mice of this batch was 10 ⁷ The bacteria/mouse (relative infection dose), i.e. lignan conditioning, improved the anti-infective resistance of the animal to the bacteria by a factor of 1000. 10 ⁷ The infectious dose of the individual bacteria is called the "relative infectious dose" and this result can be used as a reference for other infectious disease control.

The new method of the bacterial test can also explain the large-scale invasion of other pathogenic microorganisms such as new coronaviruses and the like, and the intestinal diol improves the relative infection dose of the pathogenic microorganisms.

Fig. 1 is a photograph of HE staining of the large intestine tissue of a normal mouse, and fig. 2 is a photograph of HE staining of the large intestine tissue of a mouse fed with defatted flaxseed meal group, and it was found by comparison that the large intestine tissue of a mouse fed with defatted flaxseed meal group had a large amount of lymphocyte infiltrates.

The effect of Enterodiol (END) is not only to inhibit the virus directly, but also to cause programmed viral death by stimulating the internal antiviral mechanisms of the human body, including the antiviral activity of the immune system and of the intestinal flora, thus avoiding the vicious circle of the same length as the army competition. The invention adopts a biological conversion method, namely a pure green biological manufacturing method, is harmless to the environment and can reduce the price by thousands of times. Has wide application value.

Claims

1. Application of defatted flaxseed powder in the preparation of drugs against COVID-19 infectious diseases.

2. The application of defatted flaxseed powder in the preparation of medicines against new coronavirus infectious diseases according to claim 1, characterized in that the defatted flaxseed powder is obtained by extracting flaxseed oil by low-temperature cold pressing. Defatted linseed meal is ground into fine powder of 80 to 120 mesh.

3. The application of defatted flaxseed powder in the preparation of medicines against new coronavirus infectious diseases according to claim 1, characterized in that the content of defatted flaxseed powder in the medicine against new coronavirus infectious diseases is 0.01~ 99.9wt.%.

4. The application of defatted flaxseed powder in the preparation of anti-COVID-19 infectious disease drugs according to claim 1, characterized in that the content of defatted flaxseed active pharmaceutical ingredients in the anti-COVID-19 infectious disease drugs is 0.01 ~99.9wt.%.

5. The application of defatted flaxseed powder in the preparation of anti-COVID-19 infectious disease drugs according to claim 4, characterized in that the extraction method of the defatted flaxseed effective pharmaceutical ingredients is: defatted flaxseed powder and pure water The mass ratio of defatted flaxseed powder is 1:5~10, add defatted flaxseed powder to pure water, heat to 100°C and boil for 2 minutes. The obtained water extract or the water extract obtained by drying the obtained water extract is the effective drug of defatted flaxseed. Element.

6. The application of defatted flaxseed powder according to any one of claims 2 to 5 in the preparation of drugs against novel coronavirus infectious diseases, characterized in that the drug against novel coronavirus infectious diseases also contains pharmaceutically acceptable carrier or excipient.

7. The application of defatted flaxseed powder in preparing anti-COVID-19 infectious disease drugs according to claim 6, characterized in that the pharmaceutically acceptable carrier includes liposomes, micelles, dendrimers, Microspheres or microcapsules.

8. The application of defatted flaxseed powder in the preparation of drugs against novel coronavirus infectious diseases according to claim 7, characterized in that the dosage forms of the drugs against novel coronavirus infectious diseases are tablets, granules, and capsules. , solution, pill, liposome preparation or nanoparticle preparation.

9. The application of defatted flaxseed powder in the preparation of drugs against novel coronavirus infectious diseases according to claim 8, characterized in that the drug against novel coronavirus infectious diseases is administered orally, and the oral dosage per time is Take 40g of defatted flaxseed powder and take it orally 1 to 2 times a day.