CN101869680B - New application of composition for nourishing kidney, tonifying energy, nourishing brain and allaying excitement - Google Patents
New application of composition for nourishing kidney, tonifying energy, nourishing brain and allaying excitement Download PDFInfo
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Abstract
The invention relates to application of a Chinese medicinal composition for nourishing kidney, tonifying energy, nourishing brain and allaying excitement in a medicament for treating cerebral arteriosclerosis diseases. The medicinal composition is mainly prepared from the following raw materials: ginseng, medlar, cherokee rose fruit, sealwort, Chinese dodder seed, glossy privet fruit, white peony root, epimedium herb and the like. A preparation method for the composition comprises extraction, concentration, alcohol precipitation and filtration, decolorization and decarbonization, solution preparation and other steps.
Description
The present invention is for dividing an application, and the original bill application number is 200810104844.5, and the original bill applying date is on April 24th, 2008, and the original bill name is called compositions and the preparation method and the method for quality control of kidney nourishing and replenishing vital essence, cerebral tonic tranquillizing.
Technical field
The present invention relates to a kind of Chinese medicine composition and preparation method thereof and method of quality control, particularly relate to a kind of Chinese medicine composition and preparation method thereof and method of quality control with kidney nourishing and replenishing vital essence, cerebral tonic tranquillizing.
Background technology
The people enters person in middle and old age's period, spirit and feel that animally energy lacks, and face's wrinkle begins to increase, wan and sallow not damp, dizziness and blurred vision is often arranged, Hiccough and deaf, Mental fatigue is forgetful, and melancholy nervous, frigidity and dribble of urine wait symptom not to the utmost, these symptoms, a lot of in suffering from a deficiency of the kidney, kidney-jing deficiency is the brains deficiency then, then All kinds of diseases and ailments break out for the brains deficiency, and people need the energy tonifying the kidney to consolidate the essence for operate as normal, improve body function, the curative of slow down aging.
The invention provides a kind of tonifying the kidney to consolidate the essence, improve body function, the medicine of slow down aging.
Summary of the invention
The object of the invention is to provide a kind of Chinese medicine composition with kidney nourishing and replenishing vital essence, cerebral tonic tranquillizing;
The object of the invention also is to provide a kind of Chinese medicine composition preparation method with kidney nourishing and replenishing vital essence, cerebral tonic tranquillizing;
The object of the invention also is to provide a kind of method of quality control with Chinese medicine composition of kidney nourishing and replenishing vital essence, cerebral tonic tranquillizing;
The object of the invention also is to provide the application in sexual impotence, the premature ejaculation medicine in preparation treatment cerebral arteriosclerosis of a kind of Chinese medicine composition;
The object of the invention also is to provide the application in the ocular fundus arteriosclerosis medicine in preparation treatment cerebral arteriosclerosis of a kind of Chinese medicine composition;
The object of the invention also is to provide a kind of Chinese medicine composition to have the application in the serum total cholesterol medicine in the reduction cerebral arteriosclerosis in preparation;
The object of the invention also is to provide the application in the hypertension disease medicament in preparation treatment cerebral arteriosclerosis of a kind of Chinese medicine composition;
The object of the invention also is to provide the application of a kind of Chinese medicine composition in preparation treatment prostatic hyperplasia medicine.
The present invention seeks to be achieved through the following technical solutions:
Scheme 1:
Chinese medicine composition with kidney nourishing and replenishing vital essence, cerebral tonic tranquillizing of the present invention is to be made by the crude drug of following weight ratio:
Radix Ginseng 1-6 weight portion Fructus Lycii 10-30 weight portion Fructus Rosae Laevigatae meat 10-30 weight portion
Rhizoma Polygonati 10-30 weight portion Semen Cuscutae 10-20 weight portion Fructus Ligustri Lucidi 10-20 weight portion
Radix Paeoniae Alba 10-30 weight portion Herba Epimedii 15-30 weight portion.
Chinese medicine composition with kidney nourishing and replenishing vital essence, cerebral tonic tranquillizing of the present invention is that the crude drug that is preferably as follows weight ratio is made:
Radix Ginseng 2-4 weight portion Fructus Lycii 15-30 weight portion Fructus Rosae Laevigatae meat 18-30 weight portion
Rhizoma Polygonati 15-30 weight portion Semen Cuscutae 10-20 weight portion Fructus Ligustri Lucidi 10-20 weight portion
Radix Paeoniae Alba 15-30 weight portion Herba Epimedii 20-30 weight portion.
Scheme 2:
Chinese medicine composition with kidney nourishing and replenishing vital essence, cerebral tonic tranquillizing of the present invention is to be made by the crude drug of following weight ratio:
Radix Ginseng 1-6 weight portion Fructus Lycii 10-30 weight portion Fructus Rosae Laevigatae meat 10-30 weight portion
Rhizoma Polygonati 10-30 weight portion Semen Cuscutae 10-20 weight portion Fructus Ligustri Lucidi 10-20 weight portion
Radix Paeoniae Alba 10-30 weight portion Herba Epimedii 15-30 weight portion Radix Astragali 10-30 weight portion.
Chinese medicine composition with kidney nourishing and replenishing vital essence, cerebral tonic tranquillizing of the present invention is that the crude drug that is preferably as follows weight ratio is made:
Radix Ginseng 2-4 weight portion Fructus Lycii 15-30 weight portion Fructus Rosae Laevigatae meat 18-30 weight portion
Rhizoma Polygonati 15-30 weight portion Semen Cuscutae 10-20 weight portion Fructus Ligustri Lucidi 10-20 weight portion
Radix Paeoniae Alba 15-30 weight portion Herba Epimedii 20-30 weight portion Radix Astragali 15-30 weight portion.
Scheme 3:
Chinese medicine composition with kidney nourishing and replenishing vital essence, cerebral tonic tranquillizing of the present invention is to be made by the crude drug of following weight ratio:
Radix Ginseng 1-6 weight portion Fructus Lycii 10-30 weight portion Fructus Rosae Laevigatae meat 10-30 weight portion
Rhizoma Polygonati 10-30 weight portion Semen Cuscutae 10-20 weight portion Fructus Ligustri Lucidi 10-20 weight portion
Radix Paeoniae Alba 10-30 weight portion Herba Epimedii 15-30 weight portion Radix Glycyrrhizae 3-8 weight portion
Radix Astragali 10-30 weight portion Fructus Hordei Germinatus 10-30 weight portion Mel 25-45 weight portion.
Chinese medicine composition with kidney nourishing and replenishing vital essence, cerebral tonic tranquillizing of the present invention is that the crude drug that is preferably as follows weight ratio is made:
Radix Ginseng 2-4 weight portion Fructus Lycii 15-30 weight portion Fructus Rosae Laevigatae meat 18-30 weight portion
Rhizoma Polygonati 15-30 weight portion Semen Cuscutae 10-20 weight portion Fructus Ligustri Lucidi 10-20 weight portion
Radix Paeoniae Alba 15-30 weight portion Herba Epimedii 20-30 weight portion Radix Glycyrrhizae 3-8 weight portion
Radix Astragali 15-30 weight portion Fructus Hordei Germinatus 15-30 weight portion Mel 25-45 weight portion.
Chinese medicine composition with kidney nourishing and replenishing vital essence, cerebral tonic tranquillizing of the present invention is to be made by the crude drug of following weight ratio:
Radix Ginseng 1-6 weight portion Fructus Lycii 5-10 weight portion Fructus Rosae Laevigatae meat 5-10 weight portion
Rhizoma Polygonati 5-10 weight portion Semen Cuscutae 5-10 weight portion Fructus Ligustri Lucidi 5-10 weight portion
Radix Paeoniae Alba 5-10 weight portion Herba Epimedii 10-25 weight portion Radix Glycyrrhizae 1-6 weight portion
Radix Astragali 5-10 weight portion Fructus Hordei Germinatus 2-10 weight portion Mel 20-40 weight portion.
Chinese medicine composition with kidney nourishing and replenishing vital essence, cerebral tonic tranquillizing of the present invention is to be made by the crude drug of following weight ratio:
Radix Ginseng 1-4 weight portion Fructus Lycii 5-8 weight portion Fructus Rosae Laevigatae meat 5-8 weight portion
Rhizoma Polygonati 5-8 weight portion Semen Cuscutae 5-8 weight portion Fructus Ligustri Lucidi 5-8 weight portion
Radix Paeoniae Alba 5-8 weight portion Herba Epimedii 10-20 weight portion Radix Glycyrrhizae 1-4 weight portion
Radix Astragali 5-8 weight portion Fructus Hordei Germinatus 4-7 weight portion Mel 25-35 weight portion.
Chinese medicine composition with kidney nourishing and replenishing vital essence, cerebral tonic tranquillizing of the present invention is to be made by the crude drug of following weight ratio:
Radix Ginseng 4 weight portion Fructus Lycii 12 weight portion Fructus Rosae Laevigatae meat 15 weight portions
Rhizoma Polygonati 16 weight portion Semen Cuscutae 12 weight portion Fructus Ligustri Lucidi 15 weight portions
The Radix Paeoniae Alba 12 weight portion Herba Epimedii 25 weight portion Radix Glycyrrhizaes 3 weight portions
The Radix Astragali 12 weight portion Fructus Hordei Germinatus 10 weight portion Mel 30 weight portions.
The pharmaceutical composition of scheme 1,2,3 of the present invention technology adding adjuvant is routinely made clinical acceptable forms such as tablet, capsule, oral liquid, drop pill, granule; Described adjuvant comprises solvent, disintegrating agent, correctives, antiseptic, coloring agent, binding agent, lubricant, substrate etc.
The preparation method of the Chinese medicine composition oral liquid of scheme 3 of the present invention comprises the steps:
Step 1:
Radix Ginseng water or ethanol extraction 2-4 time, each time was respectively 1-3 hour, filtered, and merging filtrate gets extracting solution A; All the other Chinese crude drug water extraction 2-4 time, each time was respectively 1-3 hour, filtered, and merging filtrate gets extracting solution B;
Step 2:
Extracting solution B is adopted the vacuum concentration method, and control solution relative density is 1.05～1.15g/ml for 70-80 ℃, the reuse filter filter concentrated solution;
Step 3:
With after the concentrated solution cooling, constantly stir, and add 95% ethanol slowly, to adjust solution and become PH=8-9, ethanol content reaches 50-80%, staticly settles 48-72 hour, gets supernatant liquid filtering;
Step 4:
The active carbon that adds percentage by weight 0.3%, ebuillition of heated then, the gained destaining solution filters, and 20 ℃ of relative densities are 1.10-1.20g/ml;
Step 5:
Add Radix Ginseng extractive solution A mixing in the medicinal liquid behind the precipitate with ethanol, and add Mel again through after taking off charcoal; Make oral liquid through conventional technology.
The preparation method of Chinese medicine composition oral liquid of the present invention is preferably as follows step:
Step 1:
Radix Ginseng ethanol extraction 3 times, each time was respectively 2 hours, filtered, and merging filtrate gets extracting solution A;
All the other Chinese crude drug water extraction 2 times, each time was respectively 2 hours, 1.5 hours, filtered, and merging filtrate gets aqueous extract B;
Step 2:
Aqueous extract B adopts the vacuum concentration method, control solution relative density D=1.05～1.15g/ml (70-80 ℃), reuse filter filter concentrated solution;
Step 3:
With after the concentrated solution cooling, constantly stir, and add 95% ethanol slowly, to adjust solution and become PH=8-9, ethanol content reaches 60-75%, and the time that staticly settles is got the liquid filtration greater than after 48 hours;
Step 4:
Add 0.3% active carbon, ebuillition of heated then, the gained destaining solution, medicine juice becomes brown liquid after filtering again, little poverty of distinguishing the flavor of, but ethanol flavor, 20 ℃ of relative density D=1.10-1.20g/ml must not be arranged;
Step 5:
Add Radix Ginseng extractive solution A mixing in the medicinal liquid behind the precipitate with ethanol, and add Mel again through after taking off charcoal; Add 3 ‰ (g/g) benzoic acid or 0.25 ‰ (g/g) nipalgin, add the diluent of 0.1-0.3% (g/g) cream flavour, furnishing PH=4.0-6.0, relative density D=1.10-1.25g/ml; Medicinal liquid was boiled 30 minutes sterilization.
Pharmaceutical composition method of quality control of the present invention comprises one or more in the following assay:
Assay 1: the assay of Radix Ginseng total saponins
Step 1: drawing standard curve
Take by weighing the ginsenoside Re and add dissolve with methanol; Get this solution respectively with microsyringe, and add methanol and make each part adding volume identical, place among the test tube respectively, add 3-7% vanillin-glacial acetic acid liquid, perchloric acid, mixing, close plug is put in 60 ℃ of waters bath with thermostatic control and is heated 10-20min, takes out and cools off with current, add glacial acetic acid, shake up, leave standstill, together with blank reagent, measure its trap, replication three times, drawing standard curve in the 560nm place with spectrophotometer;
Step 2: preparation is measured
Get present composition preparation in test tube, with after the ethyl acetate defat, with water saturated n-butyl alcohol supersound extraction 4-6 time, merge extractive liquid, is used 40-60%Na respectively
2Co
3, 0.1-0.3%NaOH, 3-6%NaOH eccysis flavone is used distilled water wash then, sloughs Na
2CO
3, NaOH, standing demix is given up water, the extract water bath method, residue is standby to 2ml with dissolve with methanol and standardize solution; Make the negative control sample by above-mentioned every step; Get the sample and the negative control product that prepare and respectively develop the color and colorimetric determination by standard curve, institute's value substitution regression equation is obtained the initial value of Radix Ginseng total saponins, deducts the negative sample measured value again, actual ginsenoside content.
Assay 2: icariine assay
Condition: high performance liquid chromatograph, chromatographic column HP ODS post ф 4.6 * 100mm detects wavelength 270nm, ABS Range 0.04, mobile phase: methanol: water=50-65: 40-55, flow 1.0ml/min, post oven temperature, degree: 36 ℃
Standard curve is drawn: getting icariine and be made into the 0.15mg/ml methanol solution, get the not commensurability HPLC that injects respectively and measure under above-mentioned analysis condition, is vertical coordinate with the peak area, and sample size is that abscissa gets standard curve;
Sample determination: the accurate sample of drawing places bottle, add ethyl acetate, ultrasonic 20-40 minute, draw the supernatant with suction pipe and place evaporating dish, so re-extract is 2-4 time, extract is put in the same evaporating dish, after volatilizing, standby in the 2ml volumetric flask with dissolve with methanol washing and standardize solution, this liquid is called A liquid; The accurate A liquid of drawing is in test tube, and the accurate 8-12%NaCl solution that adds shakes up, and freezing 50-70 minute, uses filtering with microporous membrane, the filtrate collection analysis.
According to icariine content in the sample what, sampling 2～4 μ l inject HPLC and analyze (the HPLC condition is the same), calculate content by following formula then:
μg/ml＝W/V
2×V
1×a×1/V
5
In the formula: W: the weight μ g that looks into icariine in institute's injected sample that standard curve tries to achieve by the peak area of icariine in the test sample.
V
1: the sample volume μ l for preparing.
V
2: the sample volume μ l that is injected into HPLC.
V
5: the present composition formulation samples amount ml that gets.
A: extension rate.
Assay 3: the mensuration of persticide residue
GC conditions:
Chromatographic column: elasticity capillary column ф: 1.2mmI.D.＞* 30m
Carrier gas: high-purity N
2
Split ratio: 20: 1
Column temperature: 175 ℃
Detector temperature: 280 ℃
Sample size: sample 4 μ l; Standard solution 0.4 μ l.
Get present composition oral liquid 5ml, put in the bottle, add normal hexane 4-6 time, each supersound extraction 30 minutes, merge extractive liquid, adds 2% anhydrous Na in separatory funnel
2SO
4The aqueous solution jolting is left standstill, and discards water layer, to wherein adding concentrated sulphuric acid, leaves standstill after the jolting again, and layering discards lower floor's acid solution, repeats 1～2 time by this operation, purifies to lower floor's acid solution to be colourless or light yellow, adds 2%Na again
2SO
4Aqueous solution is in separatory funnel, and jolting is left standstill, and discards water layer after the layering, repeats 2～3 times by this operation, to PH6～7, concentrates normal hexane extraction liquid, adds internal standard substance Heptachlor epoxide standardize solution, adds anhydrous sodium sulfate again, leaves standstill, and gets supernatant and carries out gas Chromatographic Determination.
The present composition has good drug effect, confirm through 300 routine clinical observation result: present composition oral liquid treatment differential diagnosis in tcm loses void, the cerebral arteriosclerosis of irritability, neurasthenia, hyperplasia of prostate determined curative effect for the kidney spleen, and total obvious effective rate of its treatment cerebral arteriosclerosis is 63.33%; Total effective rate 90.83%; Treating neurasthenic total obvious effective rate is 66.94; Total effective rate is 93.39%; The curative effect that is better than matched group respectively.Total obvious effective rate of treatment hyperplasia of prostate is 42.37%; Total effective rate is 93.22%.
Present composition oral liquid has clear improvement, and the kidney spleen is thanks to empty, the effect of disarmony between the heart and kidney syndrome, and total obvious effective rate of the curative effect of cerebral arteriosclerosis card is 66.67%, and total effective rate is 92.50%; Total obvious effective rate of the curative effect of neurasthenia's card is 67.77%, and total effective rate is 94.21%; The remarkable efficient of the curative effect of hyperplasia of prostate card is 61.02%, and total effective rate is 94.92%.The curative effect of treatment group card is better than the curative effect of matched group benefit essence in age and LAOJUNXIANLINGZHI KOUFUYE.
Present composition oral liquid can obviously improve the cerebral arteriosclerosis clinical symptoms, and its effect that improves symptoms such as dizziness is better than matched group, and can improve objective indicators such as rheoencephalogram, has the effect of cholesterol reducing.Present composition oral liquid drug safety does not have obvious influence to blood and hepatic and renal function, does not see obvious toxic and side effects and untoward reaction in the observation process, and taking convenience, welcome by extensive patients.
Following experimental example and embodiment are used to further specify but are not limited to the present invention.
Experimental example 1 cerebral arteriosclerosis symptom clinical trial
(1) data and method
1 physical data
1.1 case distribution situation
Table 1 case load distribution situation
1.2 treatment and matched group sex distribution situation: two groups of sex distribution situations of cerebral arteriosclerosis compare, through x
2Check, difference does not have significance (x
2=0.008, P＞0.05); The neurasthenia, two groups of sex distribution situations compare, and difference does not also have significance meaning (x
2=0.007, P＞0.05), comparability is arranged.
1.3 treatment and matched group age distribution situation
Two groups of age distribution situations of cerebral arteriosclerosis are relatively analyzed μ=0.67, P＞0.05 through Ridit, difference does not also have the significance meaning, and two groups of age distribution situations of neurasthenia are relatively analyzed μ=0.54 through Ridit, P＞0.05, difference also do not have the significance meaning, have comparability.
1.4 treatment and matched group course of disease distribution situation:
Analyze and the t check through Ridit, cerebral arteriosclerosis and neurasthenia, hyperplasia of prostate treatment are compared with matched group course of disease situation, and difference does not have significance meaning (P＞0.05) and has comparability.
1.5 the comparison of symptom integral value before treatment and the treatment of control group:
Before cerebral arteriosclerosis and neurasthenia treatment and the treatment of control group symptom integral value relatively, through the t check, difference there are no significant meaning (P＞0.05) illustrate and treats quite with the matched group state of an illness to have comparability.
2 cases are selected and diagnostic criteria
The sick bonded mode of card is adopted in this test, selects Chinese medical discrimination empty for the kidney spleen loses, and the cerebral arteriosclerosis of lack of preservation of spirit card is a study subject.
2.1 dialectical and diagnostic criteria
2.1.1 the kidney spleen loses void, lack of preservation of spirit is demonstrate,proved dialectical standard (with reference to five editions teaching material standards of national high medical college " Diagnostics of Chinese Medicine "
2.1.2 the cerebral arteriosclerosis diagnostic criteria is according to the tentative standard of the academic meeting revision of the 3rd department of neurology in the whole nation in 1981
2.2 include the case standard in
Meet the kidney spleen and lose void, the dialectical standard of lack of preservation of spirit and Western medicine diagnose standard.
3 test methods
3.1 grouping and contrast
Adopt random packet, positive drug contrast method.Earlier according to different sick kinds patient by the prescription on individual diagnosis serial number, again according to the random packet table by 2: 1 pro rate to test group and matched group.
3.2 trial drug
Trial drug: present composition oral liquid, specification: 10ml/ props up, lot number 9503104.
Control drug: YILINGJING KOUFUYE, to produce by Jiuzhitang Pharmaceutical Factory, specification: 10ml/ props up, lot number: 950209.
3.3 therapeutic scheme
Treatment group: present composition oral liquid, each 10ml, every day 2 times.
Tester: YILINGJING KOUFUYE, each 10ml, is used for the cerebral arteriosclerosis matched group at every day 2 times.
3.4 the course of treatment: treatment was for 6 weeks the course of treatment with matched group.
4 efficacy assessment standards
4.1 the kidney spleen loses the curative effect determinate standard (adopting " primary symptom light and heavy degree scoring method " evaluation) of void, lack of preservation of spirit card
Clinic control: treatment back primary symptom integration reduces more than 91% before the treatment;
Produce effects: treatment back primary symptom integration reduces 70～91% before the treatment;
Effectively: treatment back primary symptom integration reduces 35～69% before the treatment;
Invalid: treatment back primary symptom integration reduces＜35% before the treatment.
4.2 the curative effect determinate standard of cerebral arteriosclerosis: with reference to the standard of " the clinical research guideline of new Chinese medicine treatment cerebral arteriosclerosis " formulation
Clinic control: clinical symptoms all disappears, and checks that mainly the index result is normal.
Produce effects: most of transference cure, treatment back primary symptom integration reduces more than 70% before the treatment, checks that mainly the index result is normal substantially.
Effectively: various symptoms take a favorable turn, and treatment back primary symptom integration reduces 35%～69% before the treatment, checks that mainly the index result has improvement.
Invalid: various symptoms and inspection index result all do not have improvement, or treatment back primary symptom integration reduces＜35% than treatment is preceding.
5 statistical procedures methods
Measurement data adopts the t check, and enumeration data adopts corrected X
2Check (the counting ranked data adopt Ridit to analyze).
(2) therapeutic outcome
1.1 treatment and matched group clinical efficacy and comparison thereof: see Table 2
Table 2 treatment is compared with the matched group clinical efficacy
As can be seen from Table 2: the clinical efficacy of treatment and treatment of control group cerebral arteriosclerosis is relatively analyzed through Ridit, and difference has significance meaning (μ=2.35, P＜0.05), and the curative effect of treatment group is better than matched group.
1.2 treatment and matched group therapeutic effect of syndrome and comparison thereof: see Table 3
Table 3 treatment is compared with the matched group therapeutic effect of syndrome
Treatment is compared with the curative effect that treatment of control group kidney spleen loses void, lack of preservation of spirit syndrome, analyzes through Ridit, and difference has significance meaning (μ=2.07, P＜0.05), and the curative effect of treatment group syndrome is better than matched group.
1.3 the variation and the comparison thereof of symptom integral value before and after treatment and the treatment of control group: see Table 4
The variation (X ± SD branch) of symptom integral before and after table 4 treatment and the treatment of control group
Compare * * * P＜0.01 before and after the treatment, treatment and matched group difference are relatively
△ △P＜0.05
The variation and the comparison thereof of integrated value before and after 1.4 treatment is treated with the matched group primary symptom
Treatment group and matched group kidney spleen are thanks to empty, lack of preservation of spirit is demonstrate,proved each primary symptom treatment back integration and all obviously is less than treatment preceding (p all＜0.01), and the dizziness of wherein treatment group, sexual impotence, premature ejaculation, nocturia frequency, all diseases of dribble of urine are reduced more obvious (p＜0.01) than matched group.
Integral result (X ± SD) relatively before and after table 5 treatment and the treatment of matched group primary symptom
With the preceding relatively * * of treatment * p＜0.01; Compare △ △ p＜0.05, △ △ △ p＜0.01 with matched group.
The objective indicator testing result 1.5 cerebral arteriosclerosis is correlated with
1.5.1 blood pressure
There are 19 examples, 9 routine systolic pressures to raise before the treatment among treatment group and the matched group cerebral arteriosclerosis patient respectively, 15 examples, 7 routine diastolic pressures raise, all generally reduce after the treatment, remarkable or the highly significant (p＜0.05～0.01) with comparing difference before the treatment, but compare zero difference (p all＞0.05) between group, see table 6 for details.
Blood pressure situation comparison before and after table 6 treatment and the matched group cerebral arteriosclerosis patient treatment (Kpa, X ± SD)
With the preceding relatively * * of treatment p＜0.05, * * * p＜0.01.
1.5.2 optical fundus
The person has 35 example and 25 examples respectively to show the ocular fundus arteriosclerosis through ophthalmofundoscopy among treatment group and the matched group cerebral arteriosclerosis patient before the treatment, has 11 examples, 2 examples to improve after the treatment respectively, and comparing difference not remarkable (p＞0.05) sees table 7 for details between group.
Table 7 treatment is compared with matched group cerebral arteriosclerosis patient ocular fundus arteriosclerosis therapeutic outcome
X
2＝3.44，P＞0.05。
1.5.3 rheoencephalogram
There are 37 examples and 25 examples to show that through the rheoencephalogram inspection cerebrovascular tensity increases, blood vessel elasticity goes down respectively among preceding treatment group of treatment and the matched group cerebral arteriosclerosis patient, the normal again or improvement of the most of patient in treatment back.Compare there was no significant difference (p＞0.05) between group.See table 8 for details.
Rheoencephalogram change situation relatively behind table 8 treatment and the matched group cerebral arteriosclerosis patient treatment
μ＝0.933，p＞0.05.
1.5.4 serum total cholesterol
Treatment group and matched group cerebral arteriosclerosis patient have 42 examples and 30 routine patient Tch values to increase respectively before the treatment, generally reduce after the treatment, with comparing difference highly significant before the treatment (p all＜0.001).Compare there was no significant difference (p＞0.5) between group.See table 9 for details.
Serum total cholesterol comparison before and after table 9 treatment and the matched group cerebral arteries selenizing disease patient treatment (mmol/L, X ± SD)
With the preceding relatively * * of treatment * p＜0.01.
Experimental example 2 neurasthenia's clinical trials
(1) data and method
1 physical data, include case standard, test method, the course of treatment, statistical procedures method in experimental example 1.
2 efficacy assessment standards
2.1 the kidney spleen loses the curative effect determinate standard (adopting " primary symptom light and heavy degree scoring method " evaluation) of void, lack of preservation of spirit card
With experimental example 1.
2.2 neurasthenic curative effect determinate standard
With reference to The General Logistics Department of PLA's Ministry of Public Health " clinical disease diagnosis is according to curing the improvement standard ".
Recovery from illness: mental symptom and somatization disappear, and feel good, and recover sick preceding ability to work and life;
Produce effects: mental symptom and somatization disappear substantially, and treatment back primary symptom integration reduces more than 70% before the treatment, can be engaged in mentality and physical work;
Effectively: mental symptom and somatization alleviate, and treatment back primary symptom integration reduces 35%～69% before the treatment, can the short time carry out mentality and physical work;
Invalid: mental symptom and somatization do not have improvement, or treatment back primary symptom integration reduces＜35% before the treatment.
3 test methods
3.1 grouping and contrast
Adopt random packet, positive drug contrast method.Earlier according to different sick kinds patient by the prescription on individual diagnosis serial number, again according to the random packet table by 2: 1 pro rate to test group and matched group.
3.2 trial drug
Trial drug: present composition oral liquid, specification: 10ml/ props up, lot number 9503104.
Control drug: the just clear Pharmaceutical limited company of LAOJUNXIANLINGZHI KOUFUYE, White Cloud Mountain produces, and specification: 10ml/ props up, lot number: 950117.
3.3 therapeutic scheme
Treatment group: present composition oral liquid, each 10ml, every day 2 times.
Tester: LAOJUNXIANLINGZHI KOUFUYE, each 10ml, is used for neurasthenia's matched group at every day 2 times.
(2) therapeutic outcome
1 treatment and matched group clinical efficacy and comparison thereof: see Table 10
Table 10 treatment is compared with the matched group clinical efficacy
Treatment is compared with the neurasthenic clinical efficacy of treatment of control group, analyzes through Ridit, and difference has significance meaning (μ=2.05, p＜0.05), and the curative effect of treatment group is better than matched group.
2 treatments and matched group therapeutic effect of syndrome and comparison thereof: see Table 11
Table 11 treatment is compared with the matched group therapeutic effect of syndrome
Treatment is lost the curative effect empty, that lack of preservation of spirit is demonstrate,proved relatively with treatment of control group kidney spleen, analyzes through Ridit, and difference has significance meaning (μ=2.12, p＜0.05), and the curative effect of treatment group card is better than matched group.
The variation and the comparison of symptom integral value before and after 3 treatments and the treatment of control group: see Table 12
The variation (X ± SD branch) of symptom integral before and after table 12 treatment is treated with contrast is thin
Annotate: relatively preceding with treatment, compare between group * * * p＜0.01
△ △P＜0.05.
The variation and the comparison thereof of primary symptom integration before and after 4 treatments and the treatment of control group: see Table 13.
Treatment group and matched group kidney spleen lose void, lack of preservation of spirit is demonstrate,proved each primary symptom treatment back integration all than obvious minimizing the before the treatment (p all＜0.01), treatment is suitable with matched group this primary symptom integration situation of change of having a sleepless night, but the dizziness of treatment group, waist knee joint are weak, sexual impotence, premature ejaculation, nocturia frequency, all diseases of dribble of urine reduce more obvious (p＜0.05 or p＜0.01) than matched group.See table 13 for details
Integral result (X ± SD) relatively before and after table 13 treatment and the treatment of matched group primary symptom
Relatively preceding with treatment: * * p＜0.05, * * * p＜0.01; Compare between group:
△ △P＜0.05,
△ △ △P＜0.01.
5 treat and the influence of matched group to sleep electroencephalogram: treatment is all carried out sleep electroencephalogram examiner 32 examples with the treatment of control group front and back, and wherein 20 examples are organized in treatment, and matched group 12 examples (the results are shown in Table 14) compare not statistically significant (p＞0.05) between group.
Table 14 treatment and the influence (example) of matched group to sleep electroencephalogram
Experimental example 3 treatment prostatic hyperplasia clinical trials
(1) data and method
1 physical data, include case standard, test method, therapeutic scheme, the course of treatment, statistical procedures method in experimental example 1.
2 cases are selected and diagnostic criteria: the sick bonded mode of card is adopted in this test, selects Chinese medical discrimination empty for the kidney spleen loses, and the hyperplasia of prostate patient of lack of preservation of spirit card classifies study subject as.
3 dialectical and diagnostic criterias
3.1 the kidney spleen loses void, lack of preservation of spirit is demonstrate,proved dialectical standard with experimental example 1.
3.2 the standard that the hyperplasia of prostate diagnostic criteria is recommended according to 1993 the 1st edition " Urology Surgery " (Wu Jieping chief editor, science and technology publishing house in Shandong publishes), the rectal touch prostate size calibration that proposes in conjunction with Rous in 1985 and estimate heavy method diagnosis.
4 efficacy assessment standards
4.1 the kidney spleen loses the curative effect determinate standard (adopting " primary symptom light and heavy degree scoring method " evaluation) of void, lack of preservation of spirit card
With experimental example 1.
4.2 hyperplasia of prostate criterion of therapeutical effect
Clinic control: clinical symptoms all disappears, and checks that mainly the index result is normal.
Produce effects: most of transference cure, treatment back primary symptom integration reduces more than 70% before the treatment, checks that mainly the index result is normal substantially.
Effectively: clinical symptoms takes a turn for the better, and treatment back primary symptom integration reduces 35%～69% before the treatment, checks that mainly the index result has improvement.
Invalid: clinical symptoms and check result all do not have improvement, and perhaps treatment back primary symptom integration reduces＜35% before the treatment.
(2) therapeutic outcome
The clinical efficacy of 1 group treatment hyperplasia of prostate: this is organized in 59 examples, clinic control 11 examples, and produce effects 14 examples, effective 30 examples, invalid 4 examples, total obvious effective rate is 42.37%, total effective rate is 93.22%.
2 group treatment kidney spleens lose curative effect empty, the lack of preservation of spirit card: this is organized in 59 examples, clinic control 7 examples, and produce effects 29 examples, effective 20 examples, invalid 3 examples, total obvious effective rate 61.02%, total effective rate is 94.92%.
The variation of 3 group treatment front and back syndrome integrations
The variation (X ± SD branch) of syndrome integration before and after table 15 treatment
Annotate: relatively preceding with treatment, * * * p＜0.01.
Prostate digital rectal examination result relatively sees Table 16 before and after 4 treatments, and difference had the significance meaning before and after the result showed treatment, illustrates that present composition oral liquid has the effect of control glandular hyperplasia.
Prostate digital rectal examination result relatively before and after table 16 treatment
Through ultrasound diagnosis, weight of prostate all was higher than normally, generally alleviates after the treatment before weight of prostate was relatively treated before and after 5 treatments, and the front and back comparing difference has the significance meaning, sees Table 17.
Prostate B ultrasonic weight ratio (g, X ± SD) before and after table 17 treatment
6 safety evaluatios
Treatment is all no abnormal with treatment of control group front and back blood, excrement, routine urinalysis and hepatic and renal function inspection.These parameters does not all have significant difference between various comparisons.
The assay of experimental example 4 present composition preparation Radix Ginseng total saponinss adds recovery test and precision test
1, adds recovery test
Precision takes by weighing some parts of Re2～3mg, use a small amount of anhydrous alcohol solution, accurately add sample (through repeatedly measuring its ginsenoside content's) or negative control product (through repeatedly measure its ginsenoside content's background) 5ml, fully the mixing back is undertaken by the sample determination operations, records interpolation response rate result to be:
Sample Re addition mg records addition mg response rate %
Present composition preparation finished product 1 0.400 0.397 99.3
Present composition preparation finished product 2 0.400 0.398 99.5
Present composition preparation negative control product 1 0.400 0.408 102.0
Present composition preparation negative control product 2 0.430 0.421 97.9
Present composition preparation negative control product 3 0.400 0.384 96.0
2, precision test
Get 9001030 with 6 parts in lot number sample, extract, measure its result: Precision test result by top experimental technique:
The Radix Ginseng total saponins amount of recording mg/ml
Absolute deviation relative deviation % CV%
0.568 0.013 0.023
0.522 -0.033 -0.059
0.560 0.005 0.009
0.552 -0.003 -0.005
0.582 0.027 0.049
0.549 0.006 0.011
The mensuration precision of experimental example 5 Herba Epimedii glycosides, interpolation recovery test
1, precision test
With same lot number present composition preparation number, pour in the same exsiccant tool plug conical flask, shake up the back and use for the precision test.
Above-mentioned sample difference precision is drawn 8 parts of 2ml, handles and mensuration, record the result in following by the method in (six) sample determination:
Numbering is μ l/ml as a result
1 403
2 404
3 390
4 400
5 398
6 399
7 390
8 398
Average 398
S＝5.26
CV＝1.32％
2, add recovery test
Get negative sample number, pour in the same dry tool plug triangular flask and shake up.Accurate 8 parts in this sample of 2ml of drawing adds respectively after 600 μ g icariine standard substance (i.e. the icariine standard substance methanol solution of 300 μ l 1.2.0mg/ml) shake up, and handles and measures by the method in (six) sample determination, survey
Get the result in following:
The numbering addition μ g/ml amount of recording μ g/ml response rate %
1 300 296 98.4
2 300 302 100.9
3 300 304 101.3
4 300 solution loss
5 300 304 101.3
6 300 306 102.0
7 300 310 103.5
8 300 302 100.9
Average 101.2
S＝1.52
CV＝1.52％
Show this method accurately and reliably from precision test and interpolation recovery Tibetan experiment, can be used for test evaluation present composition formulation products quality and controlling of production process; This method experimental expenses is cheap, just can improve the separation icariine with short (100mm) post especially, is well suited for the daily big BT(batch testing) of factory and uses; In research process, find, as protein matter not being eliminated, and, be easy to damage chromatographic column, reduce post and imitate through ultrasonic membrane filtration.
Following embodiment all can realize the described effect of above-mentioned experimental example
The specific embodiment
Embodiment 1: oral liquid
Radix Ginseng 4g Fructus Lycii 30g Fructus Rosae Laevigatae meat 30g Rhizoma Polygonati 15g
Semen Cuscutae 20g Fructus Ligustri Lucidi 20g Radix Paeoniae Alba 15g Herba Epimedii 30g
Preparation technology:
Step l:
Radix Ginseng ethanol extraction 2 times, each time was respectively 2 hours, 1.5 hours, filtered, and merging filtrate gets alcohol extract;
All the other Chinese crude drug water extraction 2 times, each time was respectively 2 hours, 1.5 hours, filtered, and merging filtrate gets aqueous extract;
Step 2:
With aqueous extract, adopt the vacuum concentration method, control solution relative density D=1.05～1.15g/ml (75 ℃), reuse filter filter concentrated solution;
Step 3:
With after the concentrated solution cooling, constantly stir, and add 95% ethanol slowly, to adjust solution and become PH=8-9, ethanol content reaches 50-80%, best 60-75%, the time that staticly settles is got the liquid filtration greater than after 48 hours;
Step 4:
Add 0.3% active carbon, ebuillition of heated then, the gained destaining solution, medicine juice becomes brown liquid after filtering again, little poverty of distinguishing the flavor of, but ethanol flavor, relative density D=1.10-1.20g/ml (20 ℃) must not be arranged;
Step 5:
Add Radix Ginseng extractive solution's mixing in the medicinal liquid behind the precipitate with ethanol, and add Mel again through after taking off charcoal.Perhaps can also be at an amount of auxiliary agent of central adding, it is antiseptic, the diluent of deodorant tune, its percentage by weight is 0.1-0.3%, furnishing PH=4.0-6.5, the best is PH=4.0-6.0, relative density D=1.10-1.25g/ml, its antiseptic can be with benzoic acid or nipalgin, and deodorant tune is a cream flavour; Medicinal liquid was boiled 30 minutes sterilization.
Embodiment 2: granule
Radix Ginseng 3g Fructus Lycii 25g Fructus Rosae Laevigatae meat 25g
Rhizoma Polygonati 30g Semen Cuscutae 20g Fructus Ligustri Lucidi 20g
Radix Paeoniae Alba 30g Herba Epimedii 30g Radix Astragali 30g
Technology is made granule routinely.
Embodiment 3: capsule
Radix Ginseng 2g Fructus Lycii 30g Fructus Rosae Laevigatae meat 30g
Rhizoma Polygonati 15g Semen Cuscutae 20g Fructus Ligustri Lucidi 20g
Radix Paeoniae Alba 30g Herba Epimedii 30g Radix Astragali 30g
Technology is made capsule routinely.
Embodiment 4: tablet
Radix Ginseng 4g Fructus Lycii 15g Fructus Rosae Laevigatae meat 25g
Rhizoma Polygonati 25g Semen Cuscutae 15g Fructus Ligustri Lucidi 15g
Radix Paeoniae Alba 15g Herba Epimedii 30g Radix Glycyrrhizae 3g
Radix Astragali 30g Fructus Hordei Germinatus 15g
Technology is made tablet routinely.
Embodiment 5: oral liquid
Radix Ginseng 4g Fructus Lycii 12g Fructus Rosae Laevigatae meat 15g
Rhizoma Polygonati 16g Semen Cuscutae 12g Fructus Ligustri Lucidi 15g
Radix Paeoniae Alba 12g Herba Epimedii 25g Radix Glycyrrhizae 3g
Radix Astragali 12g Fructus Hordei Germinatus 10g Mel 30g
Preparation technology:
Step 1:
Radix Ginseng is with all the other Chinese crude drug water extraction 2 times, and each time was respectively 2 hours, 1.5 hours, filter, merging filtrate, aqueous extract;
Step 2:
With aqueous extract, adopt the vacuum concentration method, control solution relative density D=1.05～1.15g/ml, reuse filter filter concentrated solution;
Step 3:
With after the concentrated solution cooling, constantly stir, and add 95% ethanol slowly, to adjust solution and become (1) PH=8-9, ethanol content reaches 50-80%, best 60-75%, the time that staticly settles is got the liquid filtration greater than after 48 hours;
Step 4:
Add 0.3% active carbon, ebuillition of heated then, the gained destaining solution, medicine juice becomes brown liquid after filtering again, little poverty of distinguishing the flavor of, but ethanol flavor, relative density D=1.10-1.20g/ml (20 ℃) must not be arranged;
Step 5:
Add Radix Ginseng extractive solution's mixing in the medicinal liquid behind the precipitate with ethanol, and add Mel again through after taking off charcoal.Perhaps can also be at an amount of auxiliary agent of central adding, it is antiseptic, the diluent of deodorant tune, its percentage by weight is 0.1-0.3%, furnishing PH=4.0-6.5, the best is PH=4.0-6.0, relative density D=1.10-1.25g/ml, its antiseptic can be with benzoic acid or nipalgin, and deodorant tune is a cream flavour; Medicinal liquid was boiled 30 minutes sterilization.
Embodiment 6: oral liquid
Radix Ginseng 1-4g Fructus Lycii 5-8g Fructus Rosae Laevigatae meat 5-8g
Rhizoma Polygonati 5-8g Semen Cuscutae 5-8g Fructus Ligustri Lucidi 5-8g
Radix Paeoniae Alba 5-8g Herba Epimedii 10-20g Radix Glycyrrhizae 1-4g
Radix Astragali 5-8g Fructus Hordei Germinatus 4-7g Mel 25-35g
Preparation technology:
Step 1:
Radix Ginseng ethanol extraction 2 times, each time was respectively 2 hours, 1.5 hours, filtered, and merging filtrate gets alcohol extract A;
All the other Chinese crude drug water extraction 2 times, each time was respectively 2 hours, 1.5 hours, filtered, and merging filtrate gets aqueous extract B;
Step 2:
Aqueous extract B is adopted the vacuum concentration method, control solution relative density D=1.05～1.15g/ml, reuse filter filter concentrated solution;
Step 3:
With after the concentrated solution cooling, constantly stir, and add 95% ethanol slowly, to adjust solution and become PH=8-9, ethanol content reaches 60-75%, and the time that staticly settles is got the liquid filtration greater than after 48 hours;
Step 4:
Add 0.3% active carbon, ebuillition of heated then, the gained destaining solution, medicine juice becomes brown liquid after filtering again, little poverty of distinguishing the flavor of, but ethanol flavor, relative density D=1.10-1.20g/ml (20 ℃) must not be arranged;
Step 5:
Add Radix Ginseng extractive solution A mixing in the medicinal liquid behind the precipitate with ethanol, and add Mel again through after taking off charcoal.Perhaps can also be at an amount of auxiliary agent of central adding, it is antiseptic, the diluent of deodorant tune, its percentage by weight is 0.1-0.3%, furnishing PH=4.0-6.5, the best is PH=4.0-6.0, relative density D=1.10-1.25g/ml, its antiseptic can be with benzoic acid or nipalgin, and deodorant tune is a cream flavour; Medicinal liquid was boiled 30 minutes sterilization.
Embodiment 7: the icariine assay
Condition: the LC-4A high performance liquid chromatograph, chromatographic column HP ODS post ф 4.6 * 100mm detects wavelength 270nm, ABS Range 0.04, mobile phase: methanol: water=55: 45, flow 1.0ml/min, post oven temperature, degree: 36 ℃
Standard curve is drawn: accurately take by weighing icariine and be made into the 0.15mg/ml methanol solution, get 2 μ l (0.03 μ l), 3 μ l (0.45 μ g), 4 μ l (0.60 μ g), 5 μ l (0.75 μ g), 6 μ l (0.9 μ g) respectively, injecting HPLC under above-mentioned analysis condition measures, with the peak area is vertical coordinate, and sample size is that abscissa gets standard curve;
Sample determination: the accurate sample 2ml that draws, place 50ml tool plug triangular flask, add ethyl acetate 6ml, ultrasonic half an hour, draw the supernatant with suction pipe and place the 50ml evaporating dish, so re-extract is three times, extract is put in the same evaporating dish, at 60 ℃ (or under the room temperature
*) volatilize after, standby in the 2ml volumetric flask with dissolve with methanol washing and standardize solution.This liquid is called A liquid.The accurate A liquid 1ml that draws is in tool plug small test tube, and the accurate 10%NaCl solution 1ml that adds shakes up, and freezing 1 hour, use filtering with microporous membrane, filtrate collection is to be analyzed in tool grinding port plug small test tube.
According to icariine content in the sample what, sampling 2～4 μ l inject HPLC and analyze (the HPLC condition is the same), calculate content by following formula then:
μg/ml＝W/V
2×V
1×a×1/V
5
In the formula: W: the weight μ g that looks into icariine in institute's injected sample that standard curve tries to achieve by the peak area of icariine in the test sample.
V
1: the sample volume μ l for preparing.
V
2: the sample volume μ l that is injected into HPLC.
V
5: the present composition formulation samples amount ml that gets.
A: extension rate.
Embodiment 8: the assay of Radix Ginseng total saponins
Take by weighing ginsenoside Re 2.00mg and put in the 2ml volumetric flask, add dissolve with methanol, and be diluted to scale, shake up standby; Get this solution 30 respectively with microsyringe, 60,90,120,150ul, and add methanol and make each part adding volume be 150ul, place respectively among the tool plug test tube, accurately add 5% vanillin-glacial acetic acid liquid 0.20ml, perchloric acid 0.8ml, mixing, close plug is put in 60 ℃ of waters bath with thermostatic control and is heated 15min, take out with current cooling from the beginning, add glacial acetic acid 5.00ml, shake up, leave standstill 15min, together with the test solution blank, measure its trap, replication three times, drawing standard curve in the 560nm place with 722 type spectrophotometers;
Sample determination:
Get the 5.00ml sample in tool plug test tube, after the defat of 5ml ethyl acetate, with water saturated n-butyl alcohol supersound extraction 5 times, 5ml at every turn, merge extractive liquid, is used 50%Na respectively
2Co
3, 0.2%NaOH, the each 15ml eccysis of 4%NaOH flavone is used distilled water wash then, sloughs Na
2CO
3, NaOH, standing demix is given up water, the extract water bath method, residue is standby to 2ml with dissolve with methanol and standardize solution.Make the negative control sample by above-mentioned every step.Get sample and each 50ul of negative control product of preparing, develop the color and colorimetric determination by standard curve, institute's value substitution regression equation is obtained the initial value of Radix Ginseng total saponins, deducts the negative sample measured value again, actual ginsenoside content.
Embodiment 9: the mensuration of persticide residue
One, GC conditions
Instrument: Tianjin, island charged son of GC-9A is pounced on and is obtained detector and C-R3A digital treating meter
Chromatographic column: OV-17 elasticity capillary column ф: 1.2mmI.D.＞* 30m
Carrier gas: high-purity N
2
Split ratio: 20: 1
Column temperature: 175 ℃
Detector temperature: 280 ℃
Sample size: sample 4 μ l; Standard solution 0.4 μ l.
Two, reagent
1, normal hexane: get 300ml behind the analytical pure redistillation, be concentrated into 5ml with rotary evaporator, injection 5 μ l carry out gas Chromatographic Determination under the GC conditions of using, and except that this solvent peak, the height at other peak must not surpass 2 * 10
-11The peak height of g third body one Gamma Hexaochlorocyclohexane.
2, acetone: analytical pure redistillation
3, anhydrous sodium sulfate: analytical pure was burnt 4 hours for 650 ℃, was stored in the hermetic container standby.
4, distilled water: three water
5, sulphuric acid: top grade is pure
6, BHC standard substance, DDT standard substance are all available from State Standard Matter Research Centre, and purity is greater than 99%.
7, Heptachlor epoxide: Britain's import, Beijing commodity inspection and testing bureau provides
Three, sample pre-treatments
Get present composition embodiment 6 oral liquid 5ml, put in the triangular flask, add normal hexane 15ml 3 times, l0ml 2 times, each supersound extraction 30 minutes, merge extractive liquid, adds 2% anhydrous Na in separatory funnel
2SO
4Aqueous solution 40ml jolting is left standstill, and discards water layer, to wherein adding concentrated sulphuric acid 10ml, leaves standstill after the jolting again, and layering discards lower floor's acid solution, repeats 1～2 time by this operation, purifies to lower floor's acid solution to be colourless or light yellow, adds 40ml 2%Na again
2SO
4Aqueous solution is in separatory funnel, jolting is left standstill, discard water layer after the layering, repeat 2～3 times by this operation,, on the rotary evaporation ware, concentrate normal hexane extraction liquid with the special tool plug bottle of being with quantity tube to PH6～7, looking the residual content of sample middle peasant adds a certain amount of internal standard substance Heptachlor epoxide and is settled to 2ml, add anhydrous sodium sulfate 2～3g again, leave standstill, get supernatant and carry out gas Chromatographic Determination.
Claims (2)
1. the application of Chinese medicine composition in preparation treatment cerebral arteriosclerosis accompanied with hypertension medicine with kidney nourishing and replenishing vital essence, cerebral tonic tranquillizing, this pharmaceutical composition is to be made by following crude drug:
2. application as claimed in claim 1, it is characterized in that pharmaceutical composition routinely technology add adjuvant and make tablet, capsule, oral liquid, drop pill or granule.
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