CN111265523A - Application of berberine and ginsenoside composition in preparation of medicine for preventing and/or treating diabetes accompanied depression - Google Patents
Application of berberine and ginsenoside composition in preparation of medicine for preventing and/or treating diabetes accompanied depression Download PDFInfo
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/4353—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems
- A61K31/4375—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems the heterocyclic ring system containing a six-membered ring having nitrogen as a ring heteroatom, e.g. quinolizines, naphthyridines, berberine, vincamine
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7028—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
- A61K31/7034—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin
- A61K31/704—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin attached to a condensed carbocyclic ring system, e.g. sennosides, thiocolchicosides, escin, daunorubicin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
- A61P25/24—Antidepressants
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
- A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
Abstract
The invention belongs to the field of medicines, and particularly relates to an application of a berberine and ginsenoside composition in preparation of a medicine for preventing and/or treating diabetes mellitus associated with depression. The invention also provides a medicament for preventing and/or treating diabetes mellitus accompanied by depression. Comprises berberine and ginsenoside, and the mass ratio of the berberine to the ginsenoside is as follows: 60: 1-15: 4. the experimental study of the intervention effect of berberine and ginsenoside on rats with diabetes and depression is carried out by taking the classical antidiabetic and depression treating drugs of metformin hydrochloride and fluoxetine hydrochloride as positive controls, and the study result shows that the berberine and the ginsenoside can regulate insulin resistance, improve glucose metabolism of rats with diabetes and depression and improve depression symptoms.
Description
Technical Field
The invention belongs to the field of medicines, and particularly relates to an application of a berberine and ginsenoside composition in preparation of a medicine for preventing and/or treating diabetes mellitus associated with depression.
Background
Diabetes seriously harms human health, and according to the global Diabetes overview released newly by the International Diabetes union in 11 months in 2019, the number of people suffering from Diabetes in China reaches 1.164 billion, the people are at the first position in the world, the related medical expenses reach $ 1090 billion, the prevention and treatment work of Diabetes is not smooth [ International Diabetes Federation.IDF Diabetes-9 th edition.2019], the morbidity is obviously higher than the people with normal glucose metabolism in the Diabetes, the Diabetes accompanied with depression accelerates the course of the disease, increases the difficulty of medication and the fatality rate of the disease [ Chen S, Zhang Q, Dai G, et al. a meta analysis [ J ] Endocrine,2016,53(1):35-46], further discussing the occurrence mechanism of diabetes accompanied depression, and further searching effective intervention measures, which are always the treatment difficulty and long-term target.
The traditional diabetes treatment mainly focuses on blood sugar control and prevention and treatment of micro-vascular and macro-vascular diseases, while the biological-psychological-social holistic concept medical model emphasizes psychological and social factors, researches show that the depression has higher morbidity in diabetic people, and researches show that 1/3 accounts for the total number of diabetic patients, the morbidity of the diabetes patients is 3 times that of the common people,
(Potyralska MM, Krawczyk AK. suppression in Patents with type 2diabetes mellitus clinical and therapeutic indications [ J ]. Wiad Lek,2007,60(9): 449-453; weinger K, Lee J.Psychoseal and psychiatric transformers of diabetes mellitis [ J ]. Nurs Clin North Am, 2006,41(4): 667-. The diabetes patients increase the risk of adult depression, and the depression can increase the occurrence of adult diabetes, and the two have a bidirectional co-morbid mechanism to form a vicious circle [ Chulna, Chen Jing bin, Radlo billo, and the like. Western medicine treatment for diabetes accompanied depression comprises conventional hypoglycemic and antidepressant treatment, relatively few medicine choices, large toxic and side effects, poor tolerance, high cost and the like, and more serious problems are that some antidepressants also have adverse reactions for inducing blood sugar disorder [ dawn wave, towering ] diabetes and depression co-disease research progress [ J ] world clinical medicine, 2013,34(6): 364-. The traditional Chinese medicine has the characteristics of multiple ways, multiple targets, multiple links and the like, can effectively control depression symptoms and blood sugar, can improve general symptoms of the whole body, and has the irreplaceable effect of western medicines.
The traditional Chinese medicine for treating diabetes and complications thereof has a long history in China [ XL Tong, L Dong, L Chen, ZZhen, Treatment of diabetes using traditional Chinese medicine: past, present future. Am J Chin Med,2012,40(5): 877-; influence of Coptis chinensis and ginseng on the expression of spontaneous type 2diabetes rat adiponectin and its receptor [ J ], Chinese and Western medicine J, 2017(37)6:699 and 703], listing Coptis chinensis and ginseng as superior products in the earliest traditional Chinese medicine classic book Shen nong Ben Cao Jing, recording that Coptis chinensis is taken for a long time to forget, and the ginseng calms spirit, calms soul and benefits intelligence. Meanwhile, researches show that the coptis chinensis or the ginseng have certain treatment effect on depression [ Gong Ming, Huang Wen ya, Dong Hui. coptis chinensis and clinical application and modern research progress of compound treatment of depression [ J ]. Chinese Hospital pharmaceutical journal, 2018, (38)13: 1441-; the research progress of Huangshi Jing, ginseng antidepression [ J ]. Shizhen national medicine, 2014, (25)1: 175-; however, reports on the treatment of diabetes accompanied depression by coptis root, ginseng extract and combination thereof are not seen at present.
Disclosure of Invention
The invention aims to provide a new application of berberine and ginsenoside medicines.
The new application of the medicine of berberine and ginsenoside provided by the invention is as follows: application of berberine and ginsenoside in preparing medicine for preventing and/or treating diabetes accompanied depression is provided.
In the above application, the berberine can be specifically: berberine hydrochloride;
the ginsenoside can be ginsenoside Rb 1.
The invention also provides a medicament for preventing and/or treating diabetes mellitus accompanied by depression.
The medicine for preventing and/or treating diabetes accompanied depression provided by the invention comprises berberine and ginsenoside,
wherein the berberine can be specifically: berberine hydrochloride;
the ginsenoside can be ginsenoside Rb 1;
the mass ratio of the berberine to the ginsenoside is as follows: 60: 1-15: 4, more specifically 15: 2.
The medicine for preventing and/or treating diabetes accompanied depression can be prepared into various oral preparations, including: oral formulations, injections, etc., wherein the oral formulation may include: capsule, tablet, granule, powder, pill, dripping pill, sustained-release preparation, oral liquid, mixture and syrup.
The medicaments in various dosage forms can be prepared according to the conventional method in the pharmaceutical field.
The inventor further gives chronic unpredictable mild stress on the basis of a diabetic rat model, duplicates a diabetic depression-associated rat model [ Zhuyuying, bear paris, Jirongjing, and the like ] morphological research on change of hippocampal neurons of the depressed rat [ J ]. Hebei combined university report medical edition, 2013,15(5):652-653], and takes a classical antidiabetic and depression treatment drug of metformin hydrochloride combined with fluoxetine hydrochloride as a positive control to carry out experimental research on the intervention effect of the berberine and ginsenoside-involved diabetic depression rat, and research results show that the berberine and ginsenoside-can regulate insulin resistance, improve the glucose metabolism of the depression rat caused by diabetes and improve the depression symptom of the depression rat.
Aiming at the high incidence rate of the depression of the diabetes at present, the western medicine adopts the conventional blood sugar reduction and antidepressant for treatment, has relatively few medicine choices, larger toxic and side effects, poor tolerance, high cost and adverse reaction for inducing blood sugar disorder, and the invention exerts the multi-target treatment advantages of the traditional Chinese medicine by the matching combination of the berberine and the ginsenoside, performs comprehensive intervention, effectively controls the blood sugar and improves the depression symptom, and has the irreplaceable effect of the western medicine.
Detailed Description
The present invention will be described below with reference to specific examples, but the present invention is not limited thereto.
The experimental methods used in the following examples are all conventional methods unless otherwise specified; reagents, biomaterials, etc. used in the following examples are commercially available unless otherwise specified.
Examples
1. Materials and methods
1.1 test drugs
Berberine hydrochloride, Shenyang first pharmaceutical Co., Ltd, northeast pharmaceutical group.
Ginsenoside Rb1, Shanghai-derived leaf Biotech Co., Ltd.
1.2 animals and groups
Sprague-Dawleg (SD) line male rats of 50, body weight (200 s 20 g), SPF/VAF grade, supplied by Experimental animals technology, Inc., Viton, Beijing. The experimental animals are raised under the condition of no specific pathogen grade, the environmental temperature is controlled to be 22-25 ℃, the humidity is 55 +/-5%, and the food and the drinking water are freely obtained in 12/12-hour light and dark circulation.
After adaptive feeding for 1 week, the animals are randomly divided into a normal feed feeding group and a high-fat feed feeding group, 10 animals are fed with common feed, 40 rats are fed with high-fat feed, after the high-fat feed is fed for 8 weeks, 30mg/kg molding is carried out by tail vein injection of streptozotocin (STZ, prepared into 1% solution in ice bath by 0.lmol/L citric acid buffer solution, pH4.4), and the normal feed group is injected with citric acid-sodium citrate buffer solution with the same volume in tail vein. After one week, the blood sugar is measured by a tail vein tail-cutting method, and the random blood sugar value is more than 16.7mmo1/L, so that the diabetic rat model is successfully copied.
The method comprises continuously applying 50 diabetic molding rats with chronic unpredictable mild stress for molding, specifically comprising 5min of 4 deg.C ice water bath, 24h fasting, 24h of no water supply, 5min of 45 deg.C thermal stimulation, 8h of noise, 1min of tail clamping, 24h of day and night reversing, 24h of 45 deg.C squirrel cage pouring, and wet padding, wherein one stimulation is adopted every day, the same stimulation is discontinuous, and the stimulation lasts for 4 weeks. The rats with diabetes accompanied with depression are randomly divided into 5 groups: the model group is administrated with equal volume of drinking water for intragastric administration, the metformin hydrochloride and fluoxetine hydrochloride group is administrated with metformin hydrochloride 0.18g/kg and fluoxetine hydrochloride 1.8mg/kg for 1/day intragastric administration after water dissolving, the berberine and ginsenoside Rb1 are divided into A, B, C three groups, the berberine and ginsenoside A group is mixed with berberine 150mg/kg and ginsenoside Rb15mg/kg for intragastric administration of the traditional Chinese medicine composition every day, the berberine and ginsenoside B group is mixed with berberine 150mg/kg and ginsenoside Rb 120 mg/kg for intragastric administration of the traditional Chinese medicine composition every day, the berberine and ginsenoside C group is mixed with berberine 150mg/kg and ginsenoside Rb140mg/kg for intragastric administration every day, and the mixture is watered for 1 time/day for intragastric administration after water dissolving.
1.3 oral glucose tolerance test:
before the experiment, the rats are fasted and not forbidden to be watered overnight, and are subjected to intragastric administration (2g/kg) of 30% glucose solution after the experiment begins. Blood is taken 30, 60, 90 and 120 minutes after the fasting and sugar water gavage respectively to test blood sugar, and the area under the glucose curve (AUC) is calculated, wherein AUC is 1/2 (fasting blood sugar value +120 minutes value) +30 minute blood sugar value +60 minute blood sugar value +90 minute blood sugar value.
1.4 Tail suspension experiment
The tail of the experimental rat is fixed by an adhesive tape, the experimental rat is hung on a bracket, the head of the experimental rat is suspended for 5 minutes in a timing mode, and the immobility time of the rat is recorded through a camera system and small animal behavioral analysis software.
1.5 sugar Water preference test
The experimental rats were fasted for 24h with water deprivation and given two bottles of 100nl of liquid: one bottle is 1% cane sugar water, and the other bottle is common water for transportation. After 1 hour, consumption was calculated by weighing the drinking bottle and calculating the animal's preference for sugar water.
1.6 open field test
Adopting an open box of 80cm multiplied by 40cm, wherein the inner side and the ground are black, the bottom surface is divided into 25 grids with equal areas by yellow lines, placing an experimental animal in the center of the bottom surface in the box, and observing the activity condition of a rat within 5 minutes by utilizing computer-aided synchronous video recording timing: maximum moving distance (maximum movement length of rat in 5 min), total moving distance (total length of rat movement track); central zone residence time (residence time of rat in central zone); number of retentions in the central zone (number of retentions in the central zone of rats). The experimental animals are all put in from the same corner, and after the evaluation experiment is finished each time, the experimental apparatus is cleaned, wiped and aired, so as to avoid the effect of predecessors.
1.7 hyperinsulinemic euglucose clamp experiment
Rats are fasted at night, 2% pentobarbital sodium is used for intraperitoneal injection and anesthesia, the right common carotid artery and the left internal or external jugular vein are intubated, short-acting insulin is infused constantly, when the blood sugar value exceeds the range of the basic value +/-0.5 mmol/L, 20% glucose is infused, the glucose infusion speed (glucose infusion rate GIR) is adjusted, the blood sugar value is controlled to be within +/-0.5 mmol/L of the basic value, if the blood sugar value exceeds the range of the basic value +/-0.5 mmol/L, the glucose infusion rate is adjusted within the shortest time according to the blood sugar value, the blood sugar is recovered to be within +/-0.5 mmol/L of the basic value, and 3 GIRs with the clamp in a steady state are averaged.
2. Statistical method
Statistical analysis was performed using SPSS 18.0 statistical software, with the mean. + -. standard deviation of each experimental data set
Mean of or + -standard error
And (4) showing. After the test is in accordance with normal distribution, statistical analysis is carried out among the same batch of groups by adopting multi-group One-Way ANOVA (One-Way ANOVA).
3. Results
3.1 comparison of area under blood glucose and glucose Curve for oral glucose tolerance test
As shown in Table 1, the blood sugar and AUC of the model group are obviously increased compared with those of the blank group, the treatment groups are obviously reduced, the area under the OGTT glucose curve can accurately reflect the sugar metabolism of the experimental rat, and the experiment shows that each treatment group has the effect of reducing the blood sugar to a certain degree. In the traditional Chinese medicine composition with different dosages, the berberine and ginsenoside B group has better effect. Therefore, the next series of experiments were performed at the B group dosage.
TABLE 1 Experimental rats OGTT blood sugar (mmol/L) and AUC (mmol min/L)
Note: AUC, model group vs blank group P <0.01, > P < 0.05; the # P <0.01 and # P <0.05 in the treatment group and model group
3.2 comparison of hyperinsulinemic euglucose clamp test results
The high insulin positive glucose clamp test is the gold standard of insulin resistance of a diabetes model, the glucose infusion rate of the model group is obviously reduced compared with that of a normal group, the insulin resistance of the model group is shown to be obvious, and the two treatment groups are adjusted upwards to different degrees (P <0.05) (see table 2).
Table 2 results of the hyperinsulinemic euglucose clamp experiments for each group
Note: model group vs blank group P <0.01, P < 0.05; the # P <0.01 and # P <0.05 in the treatment group and model group
3.3 Tail suspension experiment
The tail suspension immobility time of the blank group is (1.78 +/-0.19) s, the model group is (4.21 +/-0.53) s, the metformin + fluoxetine hydrochloride group is (2.33 +/-0.62) s, and the berberine + ginsenoside group is (2.15 +/-0.45) s. The results of the study showed that the tail overhang time was longer in the model group than in the blank group, while the metformin + fluoxetine hydrochloride group and the berberine + ginsenoside group were reduced to different degrees than in the model group (see table 3).
3.4 sweet Water preference experiment
As shown in table 3, the percentage of carbohydrate preference in the model group was significantly reduced compared to the blank group, and the two treatment groups were more up-regulated to different degrees compared to the model group.
TABLE 3 comparison of the tail suspension time of the experiment with the sugar water preference experiment results
Note: model group vs blank group P <0.01, P < 0.05; the # P <0.01 and # P <0.05 in the treatment group and model group
3.5 open field test
As shown in table 4, the maximum moving distance, total moving distance, central region retention time and central region retention times of the model groups were all reduced compared to the model groups, indicating that more animals in the model groups stayed at the corner, less movement and improved treatment groups in different degrees.
TABLE 4 comparison of the results of the open field experiments in the rats of each group
Note: model group vs blank group P <0.01, P < 0.05; the # P <0.01 and # P <0.05 in the treatment group and model group
4. Discussion of the related Art
Diabetes seriously jeopardizes human health, about 4.63 million people in 2019 of the global population of 20-79 years old suffer from diabetes, the vast majority are type 2diabetes, China is the country with the largest number of adult patients, and the trend is expected to continue to 2045 years. Diabetes mellitus is affecting people of all ages and in all regions and is one of the most rapidly growing emergencies worldwide in the 21 st century. Previous studies have focused on controlling blood glucose and preventing and treating microvascular, macrovascular and neuropathic conditions, often ignoring the mental and emotional changes they cause. On the basis of establishing a biological-psychological-social overall concept medical model, the research on diabetes is not limited to biological factors, but gradually causes the research on psychological and social factors of people [ Chuaixiaojun, Wang Lei, Chinese and Western medicine research progress of diabetes combined depression [ J ]. Heilongjiang Chinese medicine, 2018, (13)1: 100-.
Depression is the most common mood disorder in diabetic patients. Depression is a psychiatric disorder with marked and persistent mood depression as the major manifestation. There are studies showing that about 1/4 patients with type2 or type 1 Diabetes have depressive symptoms or disorders [ Anderson RJ, Freeland KE, Clouse RE, et al. the prediction of sympodifiedpressure in additives with Diabetes a meta-Analysis [ J ]. Diabetes Care,2001,24(6):1069-1078], and also studies showing that Diabetes-associated depression accounts for up to 38.35% [ AbdulRehman Arclad, Kamranyusaf Alvi. frequency of prediction in type 2Diabetes and an Analysis of prediction factors [ J. Pak Med Assoc,2016,66 (4): 425-429]. Evidence suggests that negative mood such as anxiety, depression, etc. exacerbates the diabetic condition [ de Groot M, Crick KA, Long M, et al. RR, Ma Y, Marreo DG, et al. elongated compression systems, insoluble compressive media use, and rise of the degrading Diabetes during the degrading expression program [ J ]. Diabetes Care,2008,31(3): 420-426; lustman PJ, Griffith LS, cloud RE. Defpression in additives with solvents. solvents of 5-yr follow-up study [ J ]. Diabetes Care,1988,11(8): 605-. At present, the effective treatment medicines for diabetes accompanied with depression are relatively few and have unsatisfactory curative effect. The research on the prevention and treatment of diabetes accompanied with depression by traditional Chinese medicines is underway.
The experimental study shows that the blood sugar of the model rat is obviously increased compared with the normal group, the insulin resistance is also obviously increased compared with the normal group, chronic unpredictable mild stress is continuously given, and the model rat shows the common behavior characteristics of depression such as psychomotor inhibition, interest decline, anhedonia and the like through the detection of the behavioral indexes, thereby indicating that the diabetes concomitant depression model is successfully copied.
The area under a glucose curve of an oral glucose experiment can well reflect the blood sugar level of an experimental rat, a glucose forceps experiment is a gold standard for detecting insulin resistance of a diabetes model, and the experiment result shows that the metformin + fluoxetine hydrochloride group and the berberine + ginsenoside group have the blood sugar reducing effects of different degrees, and the mechanism of the metformin + fluoxetine hydrochloride group and the berberine + ginsenoside group is related to the improvement of the insulin resistance of experimental animals. In the behavioral test related to depression, the sugar water preference test sugar water intake, namely the preference degree of a rat to sugar water, reflects the euphoria of the rat to food, belongs to one of natural reward components, and is similar to the anhedonia of depression patients. In the tail suspension experiment, the experimental animal struggles and twists the expected turning body position at the initial position of tail suspension, and then falls into the immobile state due to hopelessness. Open field experimental behavior assessment was used to detect the initiation of rats in a new and different environment, exploration of behavior, state of stress and fear of the new environment and alertness. The three experiments are classic experiments for evaluating the curative effect of depression animals, and the research shows that the two treatment groups have different curative effects, and the berberine and ginsenoside group has the effect of improving depression to a certain extent for rats with diabetes accompanied with depression.
Insulin resistance is an important pathological basis for Diabetes, and also has an important role in the onset of Diabetes-associated depression, which can affect brain tissue glucose utilization, leading to reduced neuronal excitability, slower conduction rates, and ultimately promoting the development of depression [ Dunbar J a, Reddy P, Davis-lamellise N, et. depression: an importability with metabolic syndrome a genetic depression [ J ]. Diabetes Care,2008,31: 2368-. There are studies showing that [ Duckweed, Prunus salicina, Zingiberaceae, type 2diabetes with depressive disorder patients have changes in C-reactive protein and pancreatic islet function [ J ] test medicine, 2009,24(4): 267-one 270], and insulin resistance of diabetes with depressive disorder patients is markedly worsened, and it is further speculated that this may be related to the increase of night cortisol level, disappearance of circadian rhythm, and thus reduction of insulin sensitivity in diabetes with depressive disorder individuals. The research result shows that the berberine and ginsenoside group can regulate insulin resistance, improve glucose metabolism of rats with diabetes accompanied with depression and improve depression symptoms.
Claims (5)
1. Application of berberine and ginsenoside in preparing medicine for preventing and/or treating diabetes accompanied depression is provided.
2. A medicine for preventing and/or treating diabetes accompanied with depression comprises berberine and ginsenoside.
3. The medicament of claim 2, wherein: the berberine is as follows: berberine hydrochloride; the ginsenoside is ginsenoside Rb 1.
4. The medicament according to claim 2 or 3, characterized in that: the mass ratio of the berberine to the ginsenoside is as follows: 60: 1-15: 4.
5. the medicament according to claim 2 or 3, characterized in that: the mass ratio of the berberine to the ginsenoside is as follows: 15: 2.
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| CN102166239A (en) * | 2011-04-12 | 2011-08-31 | 中国中医科学院广安门医院 | Product for preventing and/or treating diabetes |
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| CN102166239A (en) * | 2011-04-12 | 2011-08-31 | 中国中医科学院广安门医院 | Product for preventing and/or treating diabetes |
Non-Patent Citations (3)
| Title |
|---|
| 孙亚南等: "小檗碱抗抑郁的研究进展", 《药学学报》 * |
| 尚文斌等: "人参皂苷Rb1 与小檗碱配伍对糖尿病小鼠糖脂代谢的影响", 《时珍国医国药》 * |
| 李玉倩等: "人参皂苷Rb1 抗抑郁作用的动物行为学研究", 《河南医学研究》 * |
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