Summary of the invention
Purpose of the present invention just provides a kind of AIDS virus resisting HIV, improves the health-care pu'er tea of immune function of human body.
The object of the present invention is achieved like this: the health-care pu'er tea of a kind of AIDS virus resisting HIV, raising immune function of human body is characterized in that being made up of the raw material of following mass ratio:
Pu'er tea 30~50%
The stem of noble dendrobium 10~20%
Genseng 10~20%
Poria cocos 5~10%
Rhizoma Gastrodiae 10~20%
Glossy ganoderma 8~18%.
Described health-care pu'er tea makes through following method:
A, elder generation extract the stem of noble dendrobium, genseng, Poria cocos, rhizoma Gastrodiae, glossy ganoderma raw material respectively through following method: raw material pulverizing is powdered, be soaked in water 3~5 hours, and under 80 ℃ of temperature, heated 2~4 hours, filter, concentrated filtrate, concentration is 30~50% extract;
B, Pu'er tea and biology enzyme nutrient solution mixed by 1: 0.1~0.3 mass ratio after, mix with the full dose of each extract of A step gained again, wherein, the biology enzyme nutrient solution is made up of following component by mass percent:
Hydrolysising protease 0.1~0.3%
Glucose 10~35%
Fruit drink 15~30%
Water surplus;
C, B step gained mixture is packed in the case, temperature is under the 40-65 ℃ of condition in the mixture heap, ferments 20-25 days, promptly gets health-care pu'er tea;
D, step C gained fermented health-care Pu'er tea is made loose tea, fannings, cake tea, Tuo tea, sugar tea, the tea in bag of health care products, promptly get AIDS virus resisting HIV, improve the health-care pu'er tea product of immune function of human body.
It highly is rectangle or the circle or the polygon casing of 0.8-1 rice that the vanning fermentation of described C step is adopted, and carries out under the situation of case mouth capping, to improve ferment effect, satisfies the fermentation hygienic requirements.
The hydrolysising protease of described B step is a kind of in bromelain, the papain; Fruit drink is one or more in pear juice, cider, peach juice, watermelon juice, pineapple juice, mango juice, grape juice, Passiflora juice, Cantaloupe juice, Chinese flowering quince juice, orange juice, the juice of my pomegranate.
The pH value of the biology enzyme nutrient solution of described B step is 4.5~6.5.
The control of humidity in the described sweat is 10~30% to exceed with material moisture, and water content many and few are decided by the moisture of material in the water content of solar dried green tea and other raw material self and the sweat again.
Described Pu'er tea is that the solar dried green tea of big-leaf species in yunnan tea (has another name called: Pu'er raw tea).
It is 200610010625.1 in the patent No. of on January 11st, 2006 application that described health-care pu'er tea also can adopt the applicant, and name is called the method for Chinese patent of " a kind of method with the fermented by biological enzyme Pu'er tea " and layering stand ferments and get, that is:
A, the stem of noble dendrobium, genseng, Poria cocos, rhizoma Gastrodiae, glossy ganoderma raw material pulverizing is powdered, after Pu'er tea fully mixes, mixture;
B, the mixture with the A step, water and biology enzyme nutrient solution mix by following mass ratio;
1∶0.4~0.6∶0.1~0.3
Wherein, the biology enzyme nutrient solution is made up of following component by mass percent:
Hydrolysising protease 0.1~0.3%
Glucose 10~35%
Fruit drink 15~30%
Water surplus;
C, mixed above-mentioned tealeaves mixture is deposited on the stand of layering, controlling every layer of tealeaves height is 40-70cm;
D, be under the 40-65 ℃ of condition, fermented 20-25 days, promptly get health-care pu'er tea at the tea stack temperature;
E, make loose tea, fannings, cake tea, Tuo tea, sugar tea, the tea in bag of health care products, promptly get AIDS virus resisting HIV, improve the health-care pu'er tea product of immune function of human body with the existing conventional processing method.
Layering stand wherein is made of space framework and the horizontal platen that is arranged at intervals on the space framework, and horizontal platen is made as stationary platen, perhaps the drawing and pulling type movable mold; Space framework is set to rectangular frame or triangular framing or cylindrical frame or polygonal frame, at interval horizontal support member is set on the framework, places fixing or movable mold on the support member; The horizontal support member in twos on the framework or the spacing of horizontal platen are 70-85cm.
Above-mentioned gained health-care pu'er tea can be made loose tea, fannings, cake tea, Tuo tea, sugar tea, the tea in bag of compound health-care category with other drug regimens of preventing and treating AIDS.
After described health-care pu'er tea can the conventional method by prior art extracts, the conventional method that adopts prior art again was with the extract obtained oral formulations that allows on the pharmacy such as capsule, tablet, electuary, the oral liquid etc. of being prepared into.
Described health-care pu'er tea extract can be prepared into the ejection preparation that allows on the pharmacy such as parenteral solution, powder ampoule agent for injection, injection emulsion etc. by the conventional method of prior art.
Described health-care pu'er tea extract can be made compound oral administration preparation or the compound injection that allows on the pharmacy with other drug regimens of preventing and treating AIDS.
Described medicine is Chinese herbal medicine or Western medicine.
The use amount adult who adopts can be:
Directly the health-related product made from health-care pu'er tea be 5~10 grams/time, every day 1~3 time.
The health-related product made from the health-care pu'er tea extract be 0.15~0.2 gram/time, every day 1~3 time.
In the traditional Chinese medicine ingredients in the health-care pu'er tea provided by the invention, the sweet flat profit of its property of rhizoma Gastrodiae has breath wind and ends convulsion, the effect of removing obstruction in channels to relieve pain; It is warm in nature for glossy ganoderma, and bitter and puckery flavor can nourishing and body-strengthening, strengthens the body resistance to consolidate the constitution, enhances metabolism and delay senility, and the body immune system improves the resistivity of human body to disease; Its flavor of the stem of noble dendrobium is sweet, light, and cold nature can improve the SOP (super chlorine mutase) in the human body, reduces CPO (superchlorination lipid); Its property sweet and slightly bitter taste of genseng, temperature, have take off admittedly promote the production of body fluid, the effect of tranquilizing and blood nourishing; Its property slightly sweet flavor of Poria cocos, flat has eliminating dampness and diuresis, the effect of beneficial spleen and stomach antitoxic heart-soothing and sedative.
Health-care pu'er tea provided by the invention is a kind of after fermentation tea, during the fermentation except that adding the biology enzyme nutrient solution, also add glossy ganoderma, day crudefiber crop fungi material, and the drug ingedient of the tonifying Qi of refreshing oneself, make it to play beyond thought effect, this health-care pu'er tea the more important thing is that it has very strong inhibitory action to AIDS virus HIV except that the effect with Pu'er tea self, and can obviously increase human body CD4 immunocyte quantity, and have no side effect.
The specific embodiment
Embodiment 1
Get material by following mass ratio:
300 kilograms of Pu'er raw teas
200 kilograms of the stems of noble dendrobium
100 kilograms of gensengs
100 kilograms in Poria cocos
The rhizoma Gastrodiae 120 kg
180 kilograms of glossy ganodermas
Through the applicant's the method for patent of invention formerly, that is:
A, the stem of noble dendrobium, genseng, Poria cocos, rhizoma Gastrodiae, glossy ganoderma raw material pulverizing is powdered, after Pu'er tea fully mixes, mixture;
B, be that 4.5 biology enzyme nutrient solution mixes with 1 ton of mixture of A step and 0.4 ton of water and 0.1 ton of pH value, wherein the biology enzyme nutrient solution is evenly mixed at normal temperatures by following component:
0.1 kilogram of bromelain
10 kilograms of glucose
15 kilograms of ciders
74.9 kilograms in water
C, B step gained mixture is deposited on the platen on the stand with the height of every layer of 40cm;
D, stand is placed in the fermenting cellar, the tea stack temperature is 40 ℃ on the console pallet plate, ferments 25 days, promptly gets health-care pu'er tea;
E, D step gained health-care pu'er tea is made the cake tea of health care products, promptly get AIDS virus resisting HIV, improve the health-care pu'er tea product of immune function of human body with the existing conventional processing method.
Embodiment 2
Get material by following mass ratio:
500 kilograms of Pu'er raw teas
100 kilograms of the stems of noble dendrobium
170 kilograms of gensengs
50 kilograms in Poria cocos
100 kilograms in rhizoma Gastrodiae
80 kilograms of glossy ganodermas
Through the applicant's the method for patent of invention formerly, that is:
A, the stem of noble dendrobium, genseng, Poria cocos, rhizoma Gastrodiae, glossy ganoderma raw material pulverizing is powdered, after Pu'er tea fully mixes, mixture;
B, be that 6.5 biology enzyme nutrient solution mixes with 1 ton of mixture of A step and 0.6 ton of water and 0.3 ton of pH value, wherein the biology enzyme nutrient solution is evenly mixed at normal temperatures by following component:
0.3 kilogram of papain
30 kilograms of glucose
30 kilograms of ciders
39.7 kilograms in water
C, B step gained mixture is deposited on the platen on the stand with the height of every layer of 70cm;
D, stand is placed in the fermenting cellar, the tea stack temperature is 60 ℃ on the console pallet plate, ferments 22 days, promptly gets health-care pu'er tea;
E, D step gained health-care pu'er tea is made the bag tea of health care products, promptly get AIDS virus resisting HIV, improve the health-care pu'er tea product of immune function of human body with the existing conventional processing method.
Embodiment 3
Get material by following mass ratio:
400 kilograms of Pu'er teas
140 kilograms of the stems of noble dendrobium
150 kilograms of gensengs
70 kilograms in Poria cocos
150 kilograms in rhizoma Gastrodiae
90 kilograms of glossy ganodermas
The following method of process:
A, earlier the stem of noble dendrobium, genseng, Poria cocos, rhizoma Gastrodiae, glossy ganoderma raw material are extracted through following method respectively: raw material pulverizing is powdered, be soaked in water 3 hours, heating is 4 hours under 80 ℃ of temperature, filter, concentrated filtrate, concentration is 30% extract;
B, be after 5.5 biology enzyme nutrient solution mixes, to mix with the full dose of each extract of A step gained with 400 kilograms of Pu'er teas and 80 kilograms of pH values, wherein, the biology enzyme nutrient solution is made up of following component by mass percent again:
0.2 kilogram of papain
20 kilograms of glucose
25 kilograms of fruit drinks
54.8 kilograms in water
C, B step gained mixture is packed into highly is in 0.8 meter the rectangular box, and under the situation of case mouth capping, temperature is under 40 ℃ of conditions in the control mixture heap, ferments 25 days, promptly gets health-care pu'er tea;
D, C step gained health-care pu'er tea is pressed into Tuo tea with the method for prior art, perhaps extract: health-care pu'er tea is ground into Powdered with the method for following prior art, be soaked in water 3 hours, heating is 4 hours under 80 ℃ of temperature, filter, concentrated filtrate, concentration is 80% extract, after allocating an amount of liquid glucose into, make oral liquid.
Embodiment 4
Get material by following mass ratio:
320 kilograms of Pu'er teas
110 kilograms of the stems of noble dendrobium
200 kilograms of gensengs
60 kilograms in Poria cocos
200 kilograms in rhizoma Gastrodiae
110 kilograms of glossy ganodermas
The following method of process:
A, earlier the stem of noble dendrobium, genseng, Poria cocos, rhizoma Gastrodiae, glossy ganoderma raw material are extracted through following method respectively: raw material pulverizing is powdered, be soaked in water 5 hours, heating is 2 hours under 80 ℃ of temperature, filter, concentrated filtrate, concentration is 50% extract;
B, be after 6 biology enzyme nutrient solution mixes, to mix with the full dose of each extract of A step gained with 320 kilograms of Pu'er teas and 96 kilograms of pH values, wherein, the biology enzyme nutrient solution is made up of following component by mass percent again:
0.3 kilogram of bromelain
15 kilograms of glucose
30 kilograms of fruit drinks
54.7 kilograms in water
C, B step gained mixture is packed into highly is in 1 meter the round box, and under the situation of case mouth capping, temperature is under 65 ℃ of conditions in the control mixture heap, ferments 20 days, promptly gets health-care pu'er tea;
D, C step gained health-care pu'er tea is extracted with the method for following prior art: health-care pu'er tea is ground into Powdered, was soaked in water 4 hours, heating is 3 hours under 80 ℃ of temperature, filter, concentrated filtrate, the gained filter cake carried out drying after, pack in the capsule material, get capsule.
For showing that health-care pu'er tea of the present invention has the effect of AIDS virus resisting HIV, will be verified by the test data that detects below:
I, material and method
One, measures medicine
The sample employing embodiment 3 gained concentration that detect test usefulness are 80% extract.Get this extract as testing sample, and it is dissolved in the RPMI-1640 complete medium, 0.22 μ m membrane filtration degerming, 4 ℃ of preservations, storage concentration is 5.0mg/ml.
Two, reagent and solution
(1) reagent
HEPES (N-2 (2-Hydroxyothyl) piperazine-N '-(2-ethanesufonic acid), MTT (3, (4,5-dimethylthiazol-2-y1)-2,5-diphenyl tetrazolium bromide), DMF (N, N '-Dimethyl formamine), penicillin (Penicillin), streptomycin sulphate (Streptomycin sulfate), glutamine (Glutamine) are all available from Sigma company; (2-ME 2-Mercaptoethan01) is Bio-Rad company product to 2 mercapto ethanol.RPMI-1640 and newborn calf serum are Gibeo company product.
(2) culture medium
The RPMI-1640 complete medium contains 10% newborn calf serum, 2mM L-glutamine, 10mM HEPES, 50 μ M2-mercaptoethanols, 100,000IU penicillin, 100 μ g/ml streptomysins.
Three, cell and virus
The human T lymphocyte is C8166, MT-4 and HIV-1 experiment strain HIV-1
IIIBBe so kind as to give by Britain MedicalResearchCouncil.AIDS Reagent Project.All cells and virus are all cultivated with the RPMI-1640 complete medium that contains 10% calf serum.Prepare HIV-1 according to a conventional method
IIIB, titration also calculates viral TCID
50Virus is put-70 ℃ of preservations after storing the liquid packing.The frozen according to a conventional method and recovery of cell and virus.
Four, HIV-1 infectious titration
HIV-1
IIIBPress Johnson﹠amp; The described method improvement of Byington (1990) carries out titration, is summarized as follows: HIV-1 is stored liquid on 96 orifice plates, do 4 times of dilutions, and 10 gradients, 6 repeating holes of every gradient are provided with control wells 6 holes simultaneously.Every hole: add C8166 cell 50 μ l, every hole final volume is 200 μ l.37 ℃, 5%CO
2Cultivate.Added fresh RPMI-1640 complete medium 1001 μ l on the 3rd day, under inverted microscope, observed cytopathic effect (the Cytopathic Effect that HIV-1 induces in every hole on the 7th day, CPE), whether there is the formation of plasomidum (Syncytium) to determine with every hole: to press Reed﹠amp; The Muench method is calculated the TCID of virus
50(50%Tissue Culture Infection Dose).
Five, to the toxicity test of C8166 cell
4 * 10
5/ ml C8166 cell suspension 100 μ l and different drug solution mixing to be measured establish 3 repeating holes.The control wells that does not contain medicine, 37 ℃, 5%CO are set simultaneously
2Cultivated 3 days, and adopted the MTT colorimetric method to detect cytotoxicity.The ELx800 ELIASA is measured the OD value, and the mensuration wavelength is 595nm, and reference wavelength is 630 nm.Calculate CC
50Value (50%Cytotoxic Concentration), the drug concentration during promptly to 50% normal T lymphocytic series C8166 toxigenicity.
Six, to HIV-1
IIIBThe inhibition experiment of cytopathogenic effect (CPE)
With 8 * 10
5/ ml C8166 cell 50 μ l/ holes are inoculated on the 96 porocyte culture plates that contain 100 μ l/ hole gradient doubling dilution medicines, add the HIV-1 of 50 μ l then
IIIBThe dilution supernatant, 1300 TCID
50/ hole.If 3 repeating holes.The normal cell control wells that does not contain medicine is set simultaneously.The positive medicine contrast of AZT.37 ℃, 5%CO
2Cultivated 3 days, (100 *) count plasmodial formation under the inverted microscope.EC
50(50%EffectiveConcentration) form 50% o'clock drug concentration for suppressing plasomidum.
Seven, to HIV-1
IIIBInfect the protection experiment of MT-4 cell death
On 96 porocyte culture plates, every kind of medicine is carried out 5 times of doubling dilutions, each dilution factor is established 4 holes, totally 6 dilution factors.Every hole 100 μ l.Not infection or the infected by HIV-1 that does not contain medicine is set simultaneously
IIIBThe cell control group.AZT is as the positive control medicine.2 holes in each dilution factor drip MT-4 cell suspension and HIV-1 respectively
IIIBEach 50 μ l of supernatant.2 holes drip MT-4 cell suspension 100 μ l in addition.Every hole final volume is 200 μ l.Put 37 ℃, 5%CO
2Cultivate in the incubator.Infect the back and added 100 μ l fresh cultures on the 3rd day.Measured OD with the MTT method with the ELx800 ELIASA on the 5th day
595/630The nm value.Calculate the inhibiting rate of medicine cell growth and to HIV-1
IIIBThe protective rate of infection cell.CC
50For to the toxigenous drug concentration of 50% host cell; EC
50For protecting 50% HIV-1
IIIBDrug concentration during the infection cell survival.
Eight, computing formula
Draw dose-effect curve according to experimental result, press Reed﹠amp; The Muench method calculates the 50% valid density (EC that compound suppresses virus
50), 50% suppresses cell growth concentration (CC
50) and the therapeutic index TI value (Therapeutic index) of anti-HIV-1 activity be: TI=CC
50/ EC
50
1, cell growth survival rate (%)=experimental port OD value/control wells OD value * 100
2, the cytopathogenic inhibiting rate of HIV-1 (%)=(1-experimental port plasomidum number/control wells plasomidum number) * 100
3, compound is to protective rate (%)=(experimental port OD value-HIV infection cell control wells OD value)/(normal cell control wells OD value HIV infection cell control wells 0D value) * 100 of HIV-1 infection cell
II, result
One, the cytotoxic effect of health-care pu'er tea extract
The health-care pu'er tea extract to the experimental data of the toxic action of C8166 cell see Table 1, Fig. 1.
CC
50(the health-care pu'er tea extract is tested for the first time): 378.26 μ g/ml; CC
50(the health-care pu'er tea extract is tested for the second time): 377.83 μ g/ml; CC
50(AZT) be: 1389.95 μ g/ml.
Two, the health-care pu'er tea extract is to HIV-1
IIIBInduce the cytopathic inhibitory action of C8166
The health-care pu'er tea extract is to HIV-1
IIIBInduce that the cytopathic inhibitory action experimental data of C8166 sees Table 2, Fig. 2.
EC
50(the health-care pu'er tea extract is tested for the first time): 11.62 μ g/ml; EC
50(the health-care pu'er tea extract is tested for the second time): 11.53 μ g/ml; EC
50(AZT) be: 4.01 μ g/ml
Three, the health-care pu'er tea extract is to HIV-1
IIIBInfect the protection experiment of MT-4 cell death
The health-care pu'er tea extract is to HIV-1
IIIBThe protection experimental data that infects the MT-4 cell death sees Table 3, Fig. 3.
CC
50(the health-care pu'er tea extract is tested for the first time): 93 μ g/ml; CC
50(the health-care pu'er tea extract is tested for the second time): 93.15 μ g/ml, EC
50(the health-care pu'er tea extract is tested for the first time): 19.88 μ g/ml; EC
50(the health-care pu'er tea extract is tested for the second time): 13.62 μ g/ml.CC
50(AZT)>1 μ g/ml; EC
50(AZT) be 0.0036 μ g/ml.
III, experimental result
The active settlement of exchange of health-care pu'er tea extract cytotoxicity and anti-HIV-1 summary table sees Table 4.
IV, conclusion
The health-care pu'er tea extract has certain toxicity, CC to the C8166 cell
50Be 377.83~378.26 μ g/ml, induce the C8166 cell to form plasomidum to HIV-1 and suppress better active; EC
50Be 11.53~11.62 μ g/ml, therapeutic index (TI) is 32.55~32.76.The health-care pu'er tea extract infects the MT-4 cell to HIV-1 and has certain protective role, its EC
50Be 13.62~19.88 μ g/ml, therapeutic index (TI) value is 4.68~6.84.Illustrate that this health-care pu'er tea has external anti-HIV-1 activity, its therapeutic index is 32.55~32.76.HIV-1 is infected the MT-4 cell certain protection effect is arranged.
For showing that health-care pu'er tea of the present invention has the curative effect of treatment AIDS virus HIV, below will be by its effect of clinical observation:
One, physical data
Selection meets patient's 80 examples of country's free AIDS virus resisting treatment medical science inclusion criteria, wherein, the male sex's 60 examples, women's 20 examples, the age is at 19-65 between year, average 24.5 years old, at random 80 routine patients are divided into two groups, the medicine of the single clothes of control group 40 examples AIDS virus resisting, test group 40 examples add the cake tea that the embodiment of the invention 1 provides in the clothes AIDS virus resisting medicine identical with control group.Both at aspects such as clinical manifestation, sex, ages by statistics credit analyse there was no significant difference (P>0.05).
Two, methods of treatment
All patient's antiviral therapy schemes all adopt standard one line scheme, promptly national free AIDS antiviral therapy handbook (D4T+3TC+NVP or EFV), and test group adds the cake tea that the embodiment of the invention 1 provides, one time 5 gram, boiling water brews, every day 3 times, total 4 months courses of treatment.
Three, observation index
Observation index comprises clinical symptoms, sign and laboratory indexes.Understand patient's medicine toxicity situation and assessment curative effect.Monitored body weight, CBC, blood sugar, creatinine, ALT etc., and monitored CD4 per February in every month
+Cell number.
Four, result
Test group toxicity incidence is starkly lower than control group, particularly symptom of digestive tract and hepatotoxicity wind agitation incidence, and two groups of contrasts have notable difference (P>0.05), see table 1 for details.Test group CD4
+The cell count climbing is higher than control group, sees table 2 for details.
A situation arises for table 1 toxicity
Group |
Symptom of digestive tract |
Hepatotoxicity wind agitation |
Fash |
Peripheral neuritis |
Bone marrow suppression |
Test group (40 example) |
10 examples (25%) |
2 examples (5%) |
4 examples (20%) |
2 examples (5%) |
0 example |
Control group (40 example) |
24 examples (60%) |
10 examples (25%) |
6 examples (15%) |
2 examples (5%) |
0 example |
(P>0.05)
The assessment of table 2 curative effect
Group |
CD4
+The cell count climbing
|
Opportunistic infections are sent out again |
Test group (40 example) |
36 examples (90%) [the highest rising 426/mm
3]
|
0 example |
Control group (40 example) |
24 examples (70%) |
8 examples (20%) |
(P>0.05)
Five, discuss
Composition in the health-care pu'er tea can slow down the damage of immunocyte, promotes the recovery of impaired immunocyte, adjusts effects such as intestinal absorption environment, and can improve the human immune system, gastrointestinal absorption.From this case research result, health-care pu'er tea can reduce toxicity incidence, particularly symptom of digestive tract and the hepatotoxicity wind agitation that the patient takes the AIDS virus resisting medicine, improves the compliance that the patient takes medicine, thereby improves the curative effect of antiviral therapy.This test group CD4
+The cell count climbing is higher than control group.