Patent Application
20220395551
The present invention relates to the technical fields of medicaments, health-care products or food compositions, and specifically relates to a composition and applications with functions to improve the diversity of personalized intestinal flora.
As medical innovation cutting edge of the post-antibiotic era, one of the most important scientific and technological achievements in the recent 10 years is to explain the mutualism, communalism, or parasitism relationships between gut microbiota (intestinal flora) and human body, as well as the important role of gut microbiota to maintain the homeostasis of physiological functions of the host, human body more and more clearly. Lots of diseases incapable of being treated by modern medical treatment, including metabolic, immune and mental/neurogenic diseases, are closely related to gut microbiota imbalance.
Gut microbiota makes up the shortages of substances and functions in human body with its diversity via a symbiotic relation with the host, human body. Human body has about 20,000-25,000 genes, and there are at least more than 3 million of genes in the microorganism symbiotic inner and outer surface. In other words, the genes possessed by human body by itself are lower than 1%, while the genes of microorganisms living inside and outside the surface of human body account for more than 99%. Gut microbiota is the largest community, and the vast types of genes provide extensive auxiliary functions for the host human body. The usefulness to human health can be simply generalized into: providing nutrition, helping to defense the invasion of harmful extraneous pathogens, accelerating the maturity of immune system, maintaining normal functions of immune system, helping to maintain the constant state of various physiological functions (including various mental/neural physiological activities), and the like. These progresses are renovating the traditional recognition to the metabolic, immune function, mental/neurogenic diseases in medicine such that the science of gut microbiota is entering into the stage of application and treatment from research level (Schimidt et al., Cell, 2018).
It is regretful that the eco-system of gut microbiota symbiotic to human body is worsening acutely. Led by antibiotics invented during the Second World War, the progress of public health, extensive use of pesticides and chemical drugs, environmental pollution, change of dietary structure and popularization of cesarean result in the crisis in inheritance and maintenance of intestinal bacteria. The diversity of gut microbiota is manifested by the existence of various types of vulnerable bacteria or minor groups. Minor groups (from phylum to species) may be in unfavorable conditions in competition and are poorly resistant to environmental factors. The impairment and even extinction of some strains functions thereof lead to change gut microbiota composition and function, or a shift in their local distribution called a state of dysbiosis. At the same time, the incidence rate of the metabolic, immune and mental/neurogenic diseases rapidly increases in multiples and even several tens of times. For example, for a typical disease caused by metabolic abnormality, type II diabetes, it was estimated that the number of patients with type II diabetes had been up to 114 million in 2017 (Epidemic and Control Status of Diabetes in China Adults). For another example, autism is originally a kind of rare disease, but it was shown in the Development Report 3 on Educational Rehabilitation of Autism in China that the number of Chinese patients had exceeded 10 million in 2019. Metabolic diseases mainly include: type II diabetes, obesity, cardiovascular diseases, cerebrovascular diseases, dyslipidemia, hypertension, hyperuricemia and the like; the immunologic diseases mainly include: food/drug/pollen allergy, eczema, type-I diabetes, irritable bowel syndrome, inflammatory bowel disease, Crohn's disease, multiple sclerosis, rheumatoid, asthma and cancer; the mental/neurogenic diseases mainly include: autism, depression, bipolar disorder, Parkinson's disease, Alzheimer's disease and the like. These diseases are related to multiple systems, but their common pathological basis may be associated with the imbalanced functions of gut microbiota. A professor of the New York University, Martin J. Blaser, has vividly called these diseases like modern epidemics in the book of Missing Microbes, and the reason for the epidemics may come from the imbalance of intestinal flora.
Gut microbiota is featured by personalization, and a healthy microecosystem is marked by diversity. Personalization refers to the specificity of intestinal bacteria kept by an individual, and is related to background factors such as, inheritance, culture and environment. Gut microbiota personalization generally forms around 3 years old and maintains the whole life basically. The similarity between two people with closest blood relationship (identical twins) is usually lower than 50%. Therefore, gut microbiota is compared to fingerprints by scientists. This is very important. The diversity of gut microbiota can be enhanced and maintained only in the premise of personalized gut microbiota, thus achieving the purpose of maintaining microecological balance.
Beneficial bacteria and harmful bacteria are the concepts prevailing in recent decades. However, the concepts are oversimplified and cannot correctly show the functions of gut microbiota and can even be misleading. Because the vast majority of intestinal bacteria are double-edged, namely, these intestinal bacteria will help the host in certain conditions, while in other conditions, these intestinal bacteria will damage the host human body. For example, Helicobacter pylori can cause gastric ulcer and gastric cancer. However, a large number of research data suggest that the disappearance of Helicobacter pylori causes substantially increased risk of multiple diseases such as, obesity, asthma, allergy and esophagus cancer (Li J. and Perez-Perez G I, 2018). For another example, Clostridium difficile is usually a kind of harmless common bacterium, but when the microecosystem of gut microbiota is out of balance due to antibiotics and the like, the abnormal increase of Clostridium difficile will cause pseudomembranous colitis which is a kind of fatal disease. Approximately 29,000 patients died within 30 days of the initial diagnosis of C. difficile in the United States (Press Release of the United States Department of Health and Human Services, Feb. 25, 2015). Therefore, all gut bacteria are doubled edged, the functional balance of the whole flora is the key to improve and maintain the diversity of gut microbiota. Academics have begun to note this point, and emphasized the beneficial roles for maintaining health condition, preventing and treating related chronic diseases by maintaining a harmonious microecosystem via the mechanism of interdependence and mutual inhibition of various intestinal bacteria (Cho and Blaser, Nature 2012).
Probiotics are utilized to improve the gut microbiota imbalance, which has a comparatively long history. The usefulness of probiotics is not denied, but there are three major limitations from the aspects of personalization and diversity. Firstly, as allochthonous bacteria, probiotics have been proved by multiple clinical trials to be not colonized inside the intestinal tract (Kristensen et al., Genome Med 2016). Secondly, the types of probiotics are limited, incapable of satisfying the requirements of diversity. Thirdly, as allochthonous bacteria, probiotics are competitors to the resident bacteria. Probiotics can hinder the rehabilitation of resident flora to be dangerous factors to break the balance of resident flora (Suez J, et al., Cell, 2018).
Lots of intestinal bacteria ferment ingredients incapable of being digested and absorbed by human body as own nutritional resources, such kind of nutritional resources are called prebiotics. Due to providing nutrients for inherent intestinal bacteria of an individual, prebiotics are theoretically superior to probiotics in the aspects of improving and maintaining personalized gut microbiota. However, up to now, studies are generally aimed at the increase of some specific beneficial bacteria, or decrease of harmful bacteria. Currently, there is lack of studies on the influences of prebiotics on the overall structure functions (Sanders M E et al., Nature Rev. 2019).
The present invention is coordinated with traditional Chinese herbs for strengthening spleen and invigorating Qi, clearing damp and promoting the circulation of blood based on traditional Chinese medicine acquired constitution theory, in combination with various kinds of gut microbiota's substrates to improve and maintain the personalization and diversity of gut microbiota, thereby solving the existing problems above.
According to retrieval and survey, there is no similar patent literature or paper. Related patent literatures surveyed in Chinese mainland and Taiwan districts are as follows: CN201510036300, CN201510344591, CN201510417997, Cn201510471077, CN201610189936, CN201810722697, CN_105505992_A, CN_105532904_A, CN_105535580_A, CN_105641521_A, CN_105670047, CN_105685868_A, CN_105707628_A, CN_105749099_A, CN_105797003_A, CN_106036856_A, CN106943423A, CN_107115364_A, CN_107468716_A, CN_107495049_A, CN_108159090_A, CN_108159201_A, CN_108685005_A, CN_108795823, CN_109988725_A, TW_201641021_A, TW_201726152_A, TW_201914432_A, CN201711033077, CN201810719798, CN_105596445_A, CN_105962086_A, CN_105779343_A, CN_105962086_A, CN_105969680_A, TW_201703778_A, TW_201914596_A, CN108741036A, CN106942724A, CN110214816A, CN108077912A.
The present invention relates to an intervention plan to improve and maintain personalized gut microbiota diversity by utilizing the theory of the foundation of acquired constitution from the traditional Chinese medicine, a composition functioning to improve personalized gut microbiota diversity formulated according to the intervention plan, as well as a medicament or food prepared by the composition.
The present invention first proves the usefulness thereof to personalized gut microbiota diversity by the gene analysis technology of gut microbiota. In the recent decades, the progress of high through-put sequencing technology has provided means for the intestinal bacteria gene analysis at molecular level, and prediction of functional and metabolic pathways. The species structure of a community can be known precisely by 16S/ITS sequencing. 16S rRNA gene is a kind of gene to encode ribosomal small subunits of prokaryote, and is the most common and most useful sign in the studies of bacterial systematics. The gene sequence includes 9 variable regions and 10 conserved regions; the conserved region sequence reflects the genetic relationship between species; the variable region sequence reflects the difference between species. The present invention performs amplification and sequencing on the variable region v4 for Operational Taxonomic Units (OTUs) annotation. PICRUSt is a tool which is developed at the earliest and used for predicting microflora functions based on the 16SrRNA gene sequence. After analyzing the composition of the gene function of the existing sequencing microbial genomes, the composition of the functional gene in a sample can be predicted by the composition of the species obtained by 16s sequencing. Different levels (1-3) of prediction can be obtained by the PICRUSt analysis based on 16SrDNA sequencing.
The gene analysis technology of gut microbiota makes significant progress, it cannot fully explain the whole picture of the gut microbiota, and it can even be said that the understanding of the gut microbiota is still the tip of the iceberg. The present invention is coordinated with the analysis on the system phenotype level, which can make up the limitation of the gene analysis technology. Bristol Stool Scale is utilized in the invention as a physiological parameter for the diversity and health status of gut microbiota. The scientific grounds are as follows: firstly, the nomenclator of probiotics, professor Tomotari Mitsuoka of Riken Institute has pointed that to observe the shape of feces and smell the undesirable odor of feces are the best indicators to judge whether gut microbiota is in health state (Professor Tomotari Mitsuoka interview article, 2015). Secondly, Bristol Stool Scale is considered to be the optimal integrated index to reflect the diversity of gut microbiota based on the analysis on one of the largest human study (Falony et al. Science 2016).
To further prove the improvement effect of the composition on body function by improving gut microbiota diversity, some representative laboratory detection indexes to some related diseases are tested in the present invention. These indexes include: improvement effects on dyslipidemia (serum total cholesterol, serum triglyceride, serum low density lipoprotein cholesterol, serum high density lipoprotein cholesterol), improvement effects on pathoglycemia (fasting blood-glucose and glucose tolerance test), improvement effects on hyperuricemia (serum uric acid), improvement effects on renal function indexes (serum creatinine, estimated glomerular filtration rate), and improvement effects on liver function indexes (serum alanine aminotransferase and serum aspartate aminotransferase).
The core of the present invention is achieving the balanced overall functions of gut microbiota by improving and maintaining the personalized gut microbiota diversity, beyond the restriction to the increase or decrease of an individual strain and the concept of useful bacteria and harmful bacteria. From the three levels of molecule, function and system, the present invention proves the usefulness of the composition in improving and maintaining personalized gut microbiota diversity via two-way adjusting effects. These effects can assist human body to achieve the purpose of strengthening the foundation of acquired constitution based on traditional Chinese medicine, namely, helping the development and growth of human body, maintaining the homeostasis of physiology system after being an adult, prevention, or treatment or adjunctive treatment of gut microbiota imbalance-related diseases. By the detection on the representative laboratory parameters, the composition of the present invention is proved in its 4th level, namely, pathological level of its beneficial effect on preventing and treating related diseases, in particular to metabolic diseases, immune system diseases, dermatological diseases, mental/neurogenic diseases by improving and maintaining the personalized gut microbiota diversity.
The present invention is achieved by the following technical solutions: a novel intervention plan is provided; the present invention applies the theory of strengthening the foundation of acquired constitution based on traditional Chinese medicine, chooses and uses traditional Chinese herbs for strengthening spleen and invigorating Qi, clearing damp and promoting the circulation of blood as well as compatible with substrates demanded by personalized gut microbiota diversity to break through the limitations of the concept of harmful bacteria and beneficial bacteria. Moreover, aiming at the functional balance and diversity of the overall gut microbiota, the present invention plays the roles in prevention, treatment, or adjunctive treatment of the diseases caused by dysbiosis. The composition shows definite two-way adjusting effects on gut microbiota and their predicted function, indicating that the composition has good regulating effects on the structure and diversity of gut microbiota. Meanwhile, the composition shows two-way adjusting effects on Bristol Stool Scale, which supports the effectiveness of the gut microbiota diversity from the physiological phenotype level. Through the representative laboratory detection on related diseases, the composition has improvement effects on abnormal of blood glucose, blood lipid, serum uric acid, renal function, and liver function, which further proves the usefulness of improving and maintaining the personalized gut microbiota diversity from pathological levels. To sum up, the present invention proves that the intervention plan and the composition have beneficial effects on improving and maintaining the personalized gut microbiota diversity from four levels of molecule, function, overall human body, and representative diseases.
The present invention is achieved by the following technical solutions: a composition based on the intervention plan, and an application thereof. Compatibility of medicines include traditional Chinese herbs for strengthening spleen and invigorating Qi, clearing damp and promoting the circulation of blood as well as various kinds of gut microbiota substrates (water-soluble and water-insoluble dietary fiber). All the traditional Chinese medicines or extracts of traditional Chinese medicine can be purchased from medical companies or manufacturers; all the products accord with the national quality standards. All the dietary fibers accord with the national food-grade standards.
The present invention provides a composition functioning to improving personalized gut microbiota diversity; the composition may effectively improve the functions of gut microbiota in human body after being taken.
A composition functioning to improve personalized gut microbiota diversity includes ingredients for strengthening spleen and invigorating Qi, ingredients for clearing damp and promoting the circulation of blood, and intestinal bacteria substrates.
Preferably, the ingredients for strengthening spleen and invigorating Qi include but not limited to one or more of Panax ginseng, an extract of Panax ginseng, Panax quiquefolium, and an extract of Panax quiquefolium. Panax ginseng, the extract of Panax ginseng, Panax quiquefolium and the extract of Panax quiquefolium can reinforce vital energy, invigorate spleen for benefiting lung. The Panax ginseng includes crude Panax ginseng, or simply pretreated Panax ginseng intermediate products, or crushed or ground Panax ginseng powders or granules, or the like. The Panax quiquefolium includes crude Panax quiquefolium, or simply pretreated Panax quiquefolium intermediate products, or crushed or ground Panax quiquefolium powders or granules, or the like. The extract of Panax ginseng is a primary extract extracted from Panax ginseng by the existing extraction methods, or a simply pretreated extract; the major active ingredients are ginsenoside, as well as Panax ginseng polysaccharides, polypeptides, amino acids, volatile oils, inorganic elements, and the like; the general content is 0.0001-10%, or the like. The extract of Panax quiquefolium is a primary extract extracted from Panax quiquefolium by the existing extraction methods, or a simply pretreated extract; the major active ingredients are ginsenoside, as well as volatile oils, amino acids, polysaccharides, flavonoids, inorganic elements, and the like; the general content is 0.0001-10%, or the like. The extract of Panax ginseng or the extract of Panax quiquefolium may be extracted according the existing methods, and also may be selected from the existing products purchased from the market.
Preferably, the ingredients for clearing damp and promoting the circulation of blood include an ingredient for clearing damp and an ingredient for promoting the circulation of blood, where the ingredient for clearing damp includes but not limited to one of or a mixture of more of Wolfiporia cocos and an extract of Wolfiporia cocos; and the ingredient for promoting the circulation of blood includes but not limited to one of or a mixture of more of nattokinase, natto, and an extract of natto. The Wolfiporia cocos includes one or a combination of more of crude Wolfiporia cocos, or treated Wolfiporia cocos pieces, blocks or slices, and also includes treated Wolfiporia cocos powders or granules, or the like. The extract of Wolfiporia cocos is total extracts extracted from Wolfiporia cocos by the existing extraction methods, or separated extracts, such as, pachymaran, which may be extracted by itself or selected from the products available on the market. Nattokinase may be directly selected from the existing products available on the market, or may be also replaced by natto, a natto powder or a natto extract. The final dose of the required natto, natto powder or natto extract may be determined by the amount of the fibrin degradation unit taken in need. In this solution, nattokinase dissolves thrombus by direct and indirect ways. Nattokinase may stimulate vascular endothelial cells to produce a tissue-type plasminogen activator (t-PA). t-PA activates plasminogen into plasmin to dissolve fibrous proteins, thus dissolving thrombus directly. Meanwhile, prourokinase in human body is activated into urokinase; urokinase and t-PA co-activate plasminogen, thus indirectly dissolving thrombus.
Preferably, the intestinal bacteria substrates include but not limited to a water-soluble dietary fiber, a water-insoluble dietary fiber, or any combination thereof.
The water-soluble dietary fiber includes one or more of konjaku flour, glucomannan, crude laminarin, pectin, alginic acid, glucan, guar gum, inulin and Pericarpium citri reticulatae; or one or more of food ingredients or medicinal materials containing the above ingredients. For example, Pericarpium citri reticulatae may be selected as replacement or supplementary pectin.
Preferably, the water-insoluble dietary fiber includes one or more of cellulose, hemicellulose, and methyl cellulose in lignin. Cellulose is a kind of macromolecular polysaccharide with linearly connected glucose and contained in plant cell wall. Cellulose molecules aggregate with each other (crystallization) to form microfibers, which is characterized in that adjacent microfibers are in the same direction to form firm binding bundles, insoluble into water. Hemicellulose is generally called water-insoluble polysaccharides contained in plant cell wall besides cellulose. Hemicellulose is characterized in that various saccharides (xylose, mannose, glucose, arabinose and the like) are bound irregularly into a heterogeneous multimer. Lignocellulose is a kind of organic flocculent fibrous matter obtained by processing natural timber. For example, common water-insoluble dietary fiber includes but not limited to Ganoderma lucidum, Auricularia auricual, Tremella fuciformis, Lentinus edodes, Polyporus, Hericium erinaceus, Rhizoma gastrodiae, Sanghuangporus lonicericola, Wolfiporia cocos, Psyllium seed husk, and the like; these are representative food or traditional Chinese medicines containing high-quality hemicellulose and fiber. These materials can be purchased from the market or drugstores.
Preferably, the composition further includes one or more of Siraitia grosvenorii, an extract of Siraitia grosvenorii, almond, an extract of almond, perilla seed, and an extract of perilla seed. Siraitia grosvenorii can be not only brought into spleen and large intestine channels, but also serve as a natural and healthy sweetening agent to improve the taste of the composition at the same time. Almond, extract of almond, perilla seed, and an extract of perilla seed may be further added to the composition. The almond dietary fiber has a content greater than 10%, which is rich in unsaturated fatty acids to lubricate the intestines. Perilla seed is rich in unsaturated fatty acids (>50%) and abundant essential amino acids. That is, perilla seed may provide substrates for gut microbiota, and also has the effect of lubricating the intestines and relaxing the bowels.
In the above components, a certain component may have two functions or more, for example, Wolfiporia cocos or extract of Wolfiporia cocos has the effects of strengthening spleen and clearing damp and thus, serves as a ministerial drug of the composition, and meanwhile, has more than 90% water-insoluble dietary fiber and thus, has adjuvant effects. For example, for psyllium seed husk, the component not only contains water-soluble dietary fiber, but also contains water-insoluble dietary fiber. Therefore, psyllium seed husk may both serve as the water-soluble dietary fiber and the water-insoluble dietary fiber.
As a preferred embodiment, the ingredients for strengthening spleen and invigorating Qi include one or more of Panax ginseng, the extract of Panax ginseng, Panax quiquefolium, and the extract of Panax quiquefolium; the ingredients for clearing damp and promoting the circulation of blood include an ingredient for clearing damp and an ingredient for promoting the circulation of blood, where the ingredient for clearing damp includes but not limited to one of or a mixture of more of Wolfiporia cocos and an extract of Wolfiporia cocos; and the ingredient for promoting the circulation of blood includes but not limited to one of or a mixture of more of nattokinase, natto, and an extract of natto; a daily dose mainly consists of:
an adding amount of the nattokinase is directly obtained by conversion according to the amount of the active ingredients in need; an adding amount of the natto or the extract of natto is calculated by the fibrin degradation unit of the nattokinase contained therein.
In this present invention, the daily dose may be taken once or twice or multiple times per day.
As a more preferred embodiment, the daily dose mainly consists of:
ginseng and Panax quiquefolium
Wolfiporia cocos and the extract of
Wolfiporia cocos
an adding amount of the nattokinase is directly obtained by conversion according to the amount of the active ingredients in need; an adding amount of the natto or the extract of natto is calculated by the fibrin degradation unit of the nattokinase contained therein.
As a more preferred embodiment, the composition further includes 0.1-3 g (further preferably, 0.3-1.5 g) Siraitia grosvenorii or 0.001-0.5 g (further preferably, 0.001-0.2 g) extract of Siraitia grosvenorii; or simultaneously or separately includes 0.25-1 g) almond and/or 0.25-1 g perilla seeds. The solution has treatment or relief effects on constipation patients.
In the above components, when Wolfiporia cocos is selected as the ingredient for clearing damp, 0.1-5 g crude Wolfiporia cocos need to be added; when the extract of Wolfiporia cocos is selected, 0.001-2 g extract of Wolfiporia cocos need to be added.
The water-soluble dietary fiber and the water-insoluble dietary fiber are further respectively and separately preferred as follows: the water-soluble dietary fiber accounts for 20%-60% by the total dose of the formula; the water-insoluble dietary fiber accounts for 20%-60% by the total dose of the formula. Calculated by the daily dose, the water-soluble dietary fiber or the water-insoluble dietary fiber has an adding amount of 1-10 g.
As a specific preferred embodiment, calculated by the daily dose, 0.1-1.2 g Panax ginseng and/or Panax quiquefolium may be selected as the ingredient for strengthening spleen and invigorating Qi; or 0.005-0.05 g extract of Panax ginseng and/or extract of Panax quiquefolium may be selected, or a compound of the above two solutions may be selected.
As a specific preferred embodiment, calculated by the daily dose, 0.2-4 g Wolfiporia cocos or 0.001-2 g extract of Wolfiporia cocos may be selected as the ingredient for clearing damp; 500-4000 Fu nattokinase or 10-100 g fresh natto may be selected as the ingredient for promoting the circulation of blood; further preferably, 10-50 g, or 5-50 g dry natto powder, or a crude extract of natto may be selected. Wolfiporia cocos and/or extract of Wolfiporia cocos may be not only used as a ministerial drug to assist the ingredient for strengthening spleen and invigorating Qi, but also may be used as a water-insoluble dietary fiber to have an adjuvant effect.
As a specific preferred embodiment, calculated by the daily dose, the composition includes the following components:
Wolfiporia cocos
Pericarpium citri reticulatae
Psyllium seed husk
Siraitia grosvenorii
As a specific preferred embodiment, calculated by the daily dose, the composition includes the following components:
Panax ginseng or extract of Panax quiquefolium
Wolfiporia cocos
Pericarpium citri reticulatae
Psyllium seed husk
Siraitia grosvenorii
The present invention provides an application of the composition of any one of the above technical solutions in the preparation of a medicament, food or health-care product functioning to improve personalized gut microbiota diversity at the same time.
The present invention provides an application of the composition of any one of the above technical solutions in prevention, or treatment, or adjunctive treatment of an gut microbiota imbalance-related disease.
The present invention provides an intervention plan to improve and maintain the gut microbiota diversity by means of the theory of the foundation of acquired constitution based on traditional Chinese medicine. The plan applies the traditional Chinese herbs for strengthening spleen and clearing damp, and promoting the circulation of blood, and is coordinated with compound intestinal bacteria substrates to improve and maintain the personalized gut microbiota diversity via two-way regulation on the gut microbiota functions.
Preferably, the medicament is a medicament for prevention, or treatment, or adjunctive treatment of a gut microbiota imbalance-related disease. The food or health-care product is a food or health-care product having a function of improving gut microbiota imbalance.
The present invention discloses an application of the composition of any one of the above technical solutions in improving personalized gut microbiota diversity at the same time. For example, the composition may be used in improving personalized gut microbiota diversity in a form of a medicament, food or a health-care product. Further, the present invention provides an application thereof in prevention, or treatment, or adjunctive treatment of an gut microbiota imbalance-related disease.
Preferably, the gut microbiota imbalance includes low diversity of types and functions of gut microbiota.
Preferably, the gut microbiota imbalance includes but not limited to relatively strong types and functional genomes of dominant gut microbiota, and relatively slow types and functional genomes of less abundant gut microbiota.
Preferably, the gut microbiota imbalance-related disease includes one or more of a metabolic disease, an immunologic disease, a disease of skin system, a mental/neurogenic disease and a digestive system disease which are associated with gut microbiota imbalance, as well as a low sex hormone-related disease.
Preferably, the metabolic dysfunction disease includes prediabetes, type-II diabetes, obesity, cardiovascular diseases, cerebrovascular diseases, hyperlipidemia, hypertension, hyperuricemia and renal dysfunction.
The disease caused by hyperuricemia mainly includes but not limited to gout, hyperuricemia-caused acute/chronic arthritis, uric acid-derived kidney stone, uric acid-caused acute or chronic renal dysfunction.
Preferably, the immunologic dysfunction disease includes food/drug/pollen allergy, eczema, type-I diabetes, irritable bowel syndrome, inflammatory bowel disease, Crohn's disease, multiple sclerosis, rheumatoid, chronic fatigue disease, asthma and cancer.
Preferably, the dermatological diseases includes atropine dermatitis, psoriasis, eczema, acne, pachylosis and skin aging which are associated with gut microbiota imbalance.
Preferably, the mental/neurogenic disease includes depression, bipolar disorder, autism, schizophrenia, fibromyalgia, insomnia, Parkinson's disease, and Alzheimer's disease.
Preferably, the digestive system disease includes constipation, diarrhea, fatty liver and liver cirrhosis.
Preferably, the low sex hormone-related disease includes developmental retardation of primary/secondary sex characters of children and adolescents, abnormality and disease associated with female physiological cycle and gestation period, osteoporosis caused by estrogen deficiency of postmenopausal female, male and female sexual hypofunction, and climacteric syndrome.
Preferably, the medicament may improve the body biodegradation to an exogenous toxic substance. The exogenous toxic substance includes pesticides, herbicides, food additives, plastic residues, rubber residues, environmental pollutants, heavy metals, and other hazardous chemical substances.
As a preferred embodiment, the exogenous toxic substance includes one or more of halobenzoic acid, halocyclohexane, halobenzene, caprolactam, 2-chloro-4-ethylamino-6-isopropylamino-1,3,5-triazine, dioxin, aminobenzoate and xylene.
The present invention proves that the composition may inhibit the dominant bacteria and functional genomes thereof, and strengthen the less abundant bacteria and functional genomes thereof, promote the convergence of body phenotype towards the intermediate type, and have good two-way regulation effects on gut microbiota. The composition may play the roles of prevention or therapeutical effects on metabolic, immune function, mental/neurogenic diseases and other gut microbiota imbalance-related diseases by improving and maintaining the diversity of personalized gut microbiota.
The compositions in Examples 1-7 are specified as follows: Panax ginseng is selected as a monarch drug of the compositions. Panax ginseng tastes sweet and a little bitter, and is warm and mild-natured. Panax ginseng is governed by the spleen, lung and heart meridians. Panax ginseng can reinforce vital energy, invigorate spleen for benefiting lung, promote the secretion of saliva and calm the nerves. Panax ginseng is selected as a representative drug for invigorating Qi and strengthening the spleen in the composition. Spleen governs movement and transformation, and is the foundation of acquired constitution; the nourishment of Qi and spleen makes body full of energy and blood, and full of vitality. Weak Qi and spleen lead to Weak movement and transformation, causing damp obstruction, Qi stagnation and blood stasis. Spleen prefers dryness to dampness, if the spleen is damp, movement and transformation are abnormal. Wolfiporia cocos tastes sweet and slight, is mild-natured. Wolfiporia cocos is governed by heart, lung, spleen and kidney channels, capable of clearing damp and promoting diuresis, strengthening spleen and calming heart. Wolfiporia cocos serves as the representative drug for strengthening spleen and clearing damp in the composition, and is a ministerial drug of the composition. Nattokinase is a kind of serine protease produced by Bacillus subtilis natto during soybean fermentation, found and named by a Japanese doctor, Hiroyuki Sumi. Nattokinase dissolves thrombus by direct and indirect ways. Nattokinase may stimulate vascular endothelial cells to produce a tissue-type plasminogen activator (t-PA). t-PA activates plasminogen into plasmin to dissolve fibrous proteins, thus dissolving thrombus directly. Meanwhile, prourokinase in human body is activated into urokinase; urokinase and t-PA co-activate plasminogen, thus indirectly dissolving thrombus. Therefore, the thrombolysis activity of nattokinase is about four times of plasmin. Nattokinase is selected as the representative drug for promoting the circulation of blood and removing stasis in the composition, and is another ministerial drug of the prescription. Compound types of gut microbiota substrates are combined in the composition. The substrates basically include different structures of water-soluble dietary fibers mainly derived from plants, including pectin, glucomannan, alginic acid and guar gum. Pectin is a kind of water-soluble dietary fiber which is abundant in fruits; galacturonic acid having carboxyl is connected with methyl galacturonicate whose carboxyl is esterified by methyl in a linear chain. Pericarpium citri reticulatae (total pectin is about 33%) rich in multiple types of pectin is selected in the composition (Akiko Kawabata, et al, Journal of Nutrition, 1974). Pericarpium citri reticulatae is warm-natured and governed by lung and spleen channels, and has the effects of regulating Qi and strengthening spleen, eliminating dampness to reduce phlegm. Glucomannan is a kind of water-soluble dietary fiber which is abundant in konjak plants. Glucomannan mainly consists of glucose and its diastereoisomers; β (1-4) bonds form a backbone chain, some form branched chains via β (1-3) or β (1-6) bonds. Alginic acid is a kind of water-soluble dietary fiber contained in brown seaweed, and is a linear polymer, divided into two types of β-D-mannuronic acid (M) and C-5 epimer α-L-guluronic acid (G) (both are carboxyl-carrying monosaccharides) (1-4). Crude laminarin is also called laminarin and laminaran, and mainly contains algin, fucoidan, fucose and other components, and formed by linking β(1-3)-glucan to some β(1-6)-glucosidic bonds, and is a neutral glucan. Guar gum is a kind of water-soluble dietary fiber derived from the seeds of annual leguminous plants cultivated in Pakistan and India; guar gum is a kind of polysaccharide composed of repetitive units; the backbone consists of mannose and the side chain consists of galactose. Psyllium seed husk contains about 90% dietary fiber, and is a mixture of insoluble and water-soluble dietary fibers. Psyllium seed husk contains about 18% heteropolysaccharide xylan, and is a kind of water-soluble dietary fiber. The above compound types of water-soluble dietary fibers may provide respective substrates demanded for different types of gut microbiota. The water-insoluble dietary fiber includes cellulose, hemicellulose, and lignin. Wolfiporia cocos is a kind of dry sclerotium of Poria cocos (Schw.) Wolf. Wolfiporia cocos not only has the effects of strengthening spleen and clearing damp and thus, serves as a ministerial drug of the composition, and but also contains more than 90% water-insoluble dietary fiber and thus, has adjuvant effects. Psyllium seed husk contains about 72% water-insoluble dietary fibers. The water-insoluble dietary fiber may provide a small amount of substrates for gut microbiota, what's more, may increase the water content of feces several times to hundreds of times, thus increasing the volume of excrement, beneficial to defecation. The above water-soluble and water-insoluble dietary fibers constitute the adjuvant drug of the composition. Siraitia grosvenorii is brought into spleen and large intestine channels, and serves as a conductant drug. The mogmside contained in the Siraitia grosvenorii is sweeter 300 times than sucrose, and featured by no energy, high sweetness, pure sweet taste and high safety, and is one of the most excellent natural and healthy sweetening agents. Moreover, Siraitia grosvenorii also plays the role of improving the taste of the composition.
Panax ginseng
Wolfiporia cocos
Pericarpium citri
reticulatae
Psyllium seed husk
Siraitia grosvenorii
The above components were crushed, sieved and mixed well to prepare a unit amount of the composition 1 in the present invention (namely, the amount of the following “per bag”). The composition may exert two-way regulation on the types and functions of gut microbiota, thus improving and maintaining personalized gut microbiota diversity. Moreover, the composition may keep healthy and have treatment or prevention effects on chronic diseases, in particular to metabolic diseases, immune diseases, mental/neurogenic diseases and other diseases incapable of being treated very well at present. A bag of the composition was brewed and taken once or twice per day with warm water.
Panax ginseng
Wolfiporia cocos
Pericarpium citri
reticulatae
Psyllium seed husk
Siraitia grosvenorii
The formula of this example may be used as an introducing prescription, or applied to the inadaptable subjects, such as, patients accompanied with continuous abdominal distension, or stomachache after taking the composition 1. The composition may exert two-way regulation on the types and functions of gut microbiota, thus improving and maintaining personalized gut microbiota diversity. Moreover, the composition may keep healthy and have treatment or prevention effects on chronic diseases, in particular to metabolic diseases, immune diseases, mental/neurogenic diseases and other diseases incapable of being treated very well at present. A bag of the composition (namely, the components and contents as shown in the above corresponding table) was brewed and taken once or twice per day with warm water.
Panax ginseng
Wolfiporia cocos
Pericarpium citri
reticulatae
Psyllium seed husk
Siraitia grosvenorii
The formula in the example is suitable for the patient whose feces is not improved obviously or improved slowly towards the intermediate type (type 4) of the Bristol Stool Scale after taking the composition 1 in Example 1. The above components were crushed, sieved and mixed well to prepare a unit amount of the composition 3 in the present invention. The composition may exert two-way regulation on the types and functions of gut microbiota, thus improving and maintaining personalized gut microbiota diversity. Moreover, the composition may keep healthy and have treatment or prevention effects on chronic diseases in particular to metabolic diseases, immune diseases, mental/neurogenic diseases and other diseases incapable of being treated very well at present. A bag of the composition was brewed and taken once to twice per day with warm water.
In this example, the total extract of Panax ginseng may be directly obtained from the extract of Panax ginseng purchased on the market, or the primary extract product of Panax ginseng.
Wolfiporia cocos
Pericarpium citri
reticulatae
Psyllium seed husk
Siraitia grosvenorii
The above components were crushed, sieved and mixed well to prepare a unit amount of the composition 4 in the present invention. The composition 4 may replace the composition 1. The composition may exert two-way regulation on the types and functions of gut microbiota, thus improving and maintaining personalized gut microbiota diversity. Moreover, the composition may keep healthy and have treatment or prevention effects on chronic diseases in particular to metabolic diseases, immune diseases, mental/neurogenic diseases and other diseases incapable of being treated very well at present. A bag of the composition was brewed and taken once or twice per day with warm water.
Wolfiporia cocos
Pericarpium citri
reticulatae
Psyllium seed husk
Siraitia grosvenorii
The formula of this example may be used as an introducing prescription, or applied to the inadaptable patients, such as, patients accompanied with continuous abdominal distension, or stomachache after taking the composition 1 provided in Example 1 or the composition 4 provided in Example 4. The composition 5 may replace the composition 2 provided in Example 2. The composition may exert two-way regulation on the types and functions of gut microbiota, thus improving and maintaining personalized gut microbiota diversity. Moreover, the composition may keep healthy and have treatment or prevention effects on chronic diseases in particular to metabolic diseases, immune diseases, mental/neurogenic diseases and other diseases incapable of being treated very well at present. A bag of the composition was brewed and taken once or twice per day with warm water.
Wolfiporia cocos
Pericarpium citri
reticulatae
Psyllium seed husk
Siraitia grosvenorii
The formula in the example is suitable for the patient whose feces is not improved obviously or improved slowly towards the intermediate type (type 4) of the Bristol Stool Scale after taking the composition 4 provided in Example 4. The composition 6 may replace the composition 3 provided in Example 3. The above components were crushed, sieved and mixed well to prepare a unit amount of the composition 6 in the present invention. The composition may exert two-way regulation on the types and functions of gut microbiota, thus improving and maintaining personalized gut microbiota diversity. Moreover, the composition may keep healthy and have treatment or prevention effects on chronic diseases in particular to metabolic diseases, immune diseases, mental/neurogenic diseases and other diseases incapable of being treated very well at present. A bag of the composition was brewed and taken once to twice per day with warm water.
The composition was specified below: almond (0.25-1 g) and perilla seed (0.25-1 g) were added to the composition as an adjuvant drug. The composition 7 is suitable for the patient suffering gut microbiota imbalance and diagnosed with constipation due to intestinal dryness based on traditional Chinese medicine (TCM). The composition 7 may be used flexibly according to TCM diagnosis. The almond dietary fiber has a content greater than 10% and the abundant unsaturated fatty acids therein has the effect of lubricating the intestines. Perilla seed is rich in unsaturated fatty acids (>50%) and abundant essential amino acids. That is, perilla seed may provide substrates for gut microbiota, and also have the effect of lubricating the intestines and relaxing the bowels for constipation due to intestinal dryness.
The above components were crushed, sieved and mixed well to prepare a unit amount of the composition 7 in the present invention. The composition 7 is suitable for constipation patients. The composition may exert two-way regulation on the types and functions of gut microbiota, thus improving and maintaining personalized gut microbiota diversity. Moreover, the composition may keep healthy and have treatment or prevention effects on chronic diseases in particular to metabolic diseases, immune diseases, mental/neurogenic diseases and other diseases incapable of being treated very well at present. A bag of the composition was brewed and taken once or twice per day with warm water.
Ingredients for strengthening spleen and invigorating Qi include but not limited to a crude drug or extract of Panax ginseng and/or Panax quiquefolium.
The ingredient for clearing damp includes but not limited to a crude drug or extract of Wolfiporia cocos.
The ingredient for promoting the circulation of blood includes but not limited to a crude drug or extract of nattokinase or natto.
The water-soluble dietary fiber includes but not limited to one or more of glucomannan, pectin, alginic acid, β-glucan and inulin. Food ingredients or medicinal materials, or the like capable of extracting the above these components are certainly available.
The water-insoluble dietary fiber includes but not limited to one or more of cellulose, hemicellulose, and lignin.
In the daily dose (weight) of the composition, the water-soluble dietary fiber and the water-insoluble dietary fiber are calculated by total weight percentage of the composition, including 0.01-2 g crude drug of Panax ginseng and Panax quiquefolium, or 0.001-0.2 g extract of Panax ginseng and Panax quiquefolium, 0.1-5 g crude drug of Wolfiporia cocos, or 0.001-2 g extract of Wolfiporia cocos, 10-50 g natto powder, or 500-4000 Fu nattokinase; 0.1-3 g crude drug of Siraitia grosvenorii, or 0.001-0.2 g extract of Siraitia grosvenorii. The water-soluble dietary fiber accounts for 10%-70% by the total dose of the formula; the water-insoluble dietary fiber accounts for 10%-70% by the total dose of the formula.
The present invention is achieved by the following technical solutions: the technical solutions of the present invention are proved from four levels of molecule (gene analysis of intestinal bacteria), function (functional genomes analysis), system phenotype (Bristol Stool Scale), and some representative laboratory detection indexes, including blood lipid (total cholesterol, triglyceride, low density cholesterol and high density cholesterol), blood glucose (fasting blood-glucose, glucose tolerance test), serum uric acid, renal function indexes (serum creatinine and estimated glomerular filtration rate), and liver function indexes (alanine aminotransferase and serum aspartate aminotransferase). The composition may improve and maintain the personalized gut microbiota diversity via two-way regulation on gut microbiota, thus preventing or treating various diseases associated with gut microbiota imbalance.
The molecular level analysis was performed by OTU clustering and species annotation. Vsearchv2.4.4 was used for (OTU) analysis; the sequence was subjected to repetitive sequence removal (--derep_fulllength), clustering (--cluster_fast,-id 0.97) and chimera removal (--uchime_ref) (Rognes2016), then clustered into OTUs according to 97% similarity by default. Default parameters were used to select an OTU representative sequence; and the representative sequence was subjected to species annotation via VSEARCH based on SILVA128 database (Quastet al. 2013) to generate an OTU list, thus obtaining the community composition in each sample at the level of Phylum, Class, Order, Family, Genus and Species, as well as the abundance and classification of all the OTUs in each sample. For the quality control of the raw sequence data, the data size of the tags and clean tags was firstly judged whether of satisfying the sequencing requirements; generally, when the data size of the tags was up to 30,000 reads above, the sequencing requirements were satisfied. Reads of the mean clean tags of the 16 s sample in this test were 107670.08, completely capable of satisfying the sequencing requirements. OTUs whose content was lower than 0.001% of the total sequences in the sample were not taken in this experiment due to the current detection sensitivity.
For the functional genome analysis, the number of Species and Genus of the raw 16s sequencing data was standardized. A comparatively mature tool PICRUSt (PICRUSt Nature Biotechnology, 1-10. 8 2013) which is developed at the earliest and used for predicting microflora functions based on 16S rRNA gene sequence was then utilized to obtain the predictive functions of the genome.
As for the system phenotype analysis, Bristol Stool Chart, as shown in
Modified Cleveland Stool Score, as an auxiliary index of defecation, is also taken in the present invention. The method may be used to achieve better comprehensive analysis on the improvement effects on constipation based on the point of Bristol Stool Scale plus indexes listed in the two tables.
The administration method, administration time, detection method and statistical treatment are concluded and specified as follows:
Moreover, it should be noted that in the table above, the subjects administered for 14 days and the subjects administered for 30 days were from the same group of patients. Feces of these subjects were tested twice to confirm the intervention effect of the composition on the subacute (14 days) and long-term (30 days) of gut microbiota. Meanwhile, repeatability of the drug effect was also verified by the comparison of the two test results.
The composition has good two-way regulation effect on Phylum, Class, Order, and Family. The so-called two-way regulation effect mainly refers that the composition may selectively enhance the relatively less abundant strain and their function. The two-way regulation effect is reflected in that the lower the relative abundance of the bacteria, or functional genomes is before administration, the higher the relative abundance of the bacteria, or functional genomes is after administration. The increase of the relatively less abundant intestinal bacteria or functional genomes proportionally lower the ratio of dominant bacteria or functional genomes, capable of strengthening and maintaining the microecological balance of gut microbiota. It will be abbreviated for two-way regulation in the following description of the present invention, and will be not repeated any more.
As shown in
The composition has an inhibitory effect on the most dominant bacterium at the level of Phylum, Bac_Firmicutes. As shown in
The composition significantly improves the abundance of the second dominant bacterium at the level of Phylum, Bac_Bacteroidetes. As shown in
The composition significantly improves the abundance of the third dominant bacterium at the level of Phylum, Bac_Proteobacteria. As shown in
The composition may improve the abundance of other less abundant bacteria at the level of Phylum. As shown in
As shown in
To sum up, the composition may inhibit the relative abundance of dominant bacteria and enhance the relative abundance of the less abundant bacteria (or relatively less abundant dominant bacteria) at the level of Phylum, and shows good two-way regulation. The enhancement of the less abundant bacteria at the level of Phylum indicates that the composition may help the reproduction and maintenance of low-abundance bacteria, thus helping improving and maintaining the diversity of gut microbiota, and keeping the health condition of the gut microbiota microecosystem.
As shown in
Gut-brain axis is an existing concept. The research progress on gut microbiota brings new contents for such a concept. For example, autism is always accompanied with digestive tract symptoms such as, constipation and partial eclipse and thus, has been concerned extensively in the correlation with gut microbiota for the past few years. The latest shocking discovery has proved that the decrease of several specific bacterial genus, including Bacteroides and Parabacteroides, and the increase of Lachnospriraceae are the reasons (causative) to cause abnormal behaviors of autism (Sharon et al. Cell 2019). The composition may significantly decrease the relative abundance of Lachnospriraceae (
Researches show that compare to the composition of bacteria, there is always a smaller inter-individual functional variation, which may reflect the condition of the gut microbiota microecosystem better. The present invention further specifies the effects of the composition from the functional level by analyzing the functional genomes of gut microbiota.
The present invention further performs in-depth studies on the usefulness of the composition from the following four aspects. Firstly, from the angle of core function of gut microbiota, namely, maintaining self-survival and reproduction by bacteria,
As shown in
The cellular process includes repair and replication functions of DNA. As shown in
As shown in
The cellular process includes cell apoptosis and protein degradation through Ubiquitin system. As shown in
Ubiquitin system is another cell protein degradation chain reaction. Ubiquitin system participates in most of the short-period protein degradation, and participates in lots of cell life processes by regulating the metabolism of intracellular proteins.
Peptidoglycan is an important component of bacterial cell wall. The composition shows certain two-way regulation on Glycan Biosynthesis. As shown in
Xenobiotics biodegradation is an important function to gut microbiota. Xenobiotics include medicaments, pesticides, plastics or rubber residues, environmental pollutants, food additives, and the like; most of them are harmful to human health. As shown in
The table below summarizes the acceleration effects of the composition on some important functional genomes of xenobiotics biodegradation. These xenobiotics include DDT raw materials, the level-I cancerogenic substance, Dioxin, and atrazine, the herbicide applied most widely in the world, and the like.
112%
As shown in
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As shown in
As shown in
As shown in
Besides the precursor as shown in
Lysine is one of the essential amino acids to human and mammal, cannot be synthesized by the body itself and has to be supplemented from the outside. Gut microbiota has the genes to synthesize lysine. As shown in
As shown in
As shown in
Gut microbiota plays a decisive role to the maturity of the host human body immune system and function maintenance. As foreign bodies, gut microbiota needs to be monitored by immune system; but as a mutualistic group, gut microbiota cannot be attacked by human immune cells. Such kind of balance actually plays the role of training body's immune system to recognize harmful and innocuous exogenous substances. Gut microbiota are vividly compared to the accelerator and brake to control the host, the body's immune system by Justin L. Sonnenberg, a professor of Stanford University. At present, the most important problem is hypofunction of the brake, the light one suffers food or pollen allergy, for the serious one, immune cells will attack tissues of the body. Lipopolysaccharide (LPS) plays a crucial role to the accelerator and brake during the interaction between gut microbiota and the host body's immune system. LPS acts on the pattern recognition receptor of Toll-like receptors (TLRs). LPS promotes the development and maturity of the host body's immune system by the TLR4 receptor existing in target cells and plays the role similar to an accelerator. Meanwhile, LPS possessed by gut microbiota also plays a crucial role in adjusting the brake functions of immune system. Such a function is mainly achieved by activating the regulatory T cells to control the activity of immune system (Belkaid Y et al., Cell 2014; Tokuhara D et al., Allergology Int. 2019). The decrease of regulatory functions will cause immunologic dysfunction, and is closely related to the attack of various allergic diseases, such as food/drug/pollen body allergy, eczema; and autoimmune diseases, such as type-I diabetes, irritable bowel syndrome, inflammatory bowel disease, Crohn's disease, multiple sclerosis, rheumatoid, asthma and cancer.
Natural immune system is an important line of defense for human body to resist pathogen invasion. Innate immunity further has RIG-I like Receptors (RLRs) besides the above pattern recognition receptor of Toll-like Receptors (TLRs). RIG-I belongs to RLRs. Interferon is triggered for expression after detecting the 5′-triphosphoric acid double-stranded RNA (5′ppp-dsRNA) in cells derived from virus, thus inhibiting the virus infection.
Bristol Stool Score is utilized in this present invention as an integrated index of gut microbiota diversity to analyze the effect of the composition from the system level. It can be seen from observation in human body that the composition has significant two-way regulation. As shown in
The Cleveland Stool Score improved on the basis of the Bristol Stool Score also shows obvious improvement effects. As shown in
To further prove the improvement effect of the composition on body functions, representative laboratory detection indexes directed to some volunteers suffering from hyperglycemia, dyslipidemia or high serum uric acid were detected. These indexes include blood lipid (serum total cholesterol, serum triglyceride, serum low density lipoprotein cholesterol, serum high density lipoprotein cholesterol), blood glucose (fasting blood-glucose and glucose tolerance test), serum uric acid, renal function indexes (serum creatinine, estimated glomerular filtration rate), and liver function indexes (serum alanine aminotransferase and serum aspartate aminotransferase).
The administration method was the same as that for the volunteers with gut microbiota detection. Administration method is briefly specified as follows: 1) the composition 1 or 4 was taken first (in this example, the subsequent test data was obtained on the condition that the composition 1 was mixed with 250-300 ml warm water and taken twice per day as the starting formula). After taking the composition, some of patients would suffer abdominal distension and even stomachache, and the symptoms of abdominal distension and stomachache would be improved or removed within two or three days generally, and then, the composition may be continuously taken. 2) If the reaction of abdominal distension and even stomachache last a week around, the composition 2 or 5 was used for replacement (in this example, the subsequent test data was obtained on the condition that the composition 2 was taken as the adjustment formula). After the patient adapted to the composition one week later, if the condition of the feces (Bristol Stool Scale) was good, the composition 2 or 5 may serve as the maintenance prescription. If the condition of feces (Bristol Stool Scale) had no obvious response, the composition 1 or 4 was taken again once the abdominal distension or stomachache was improved or removed. 3) If the condition of feces (Bristol Stool Scale) had no obvious response one week later after taking the composition 1 or 4, the composition 3 or 6 was used (in this example, the subsequent test data was obtained on the condition that the composition 3 was taken as the adjustment formula). If the condition of feces (Bristol Stool Scale) had good response, the composition 1 or 4 was taken again as the maintenance prescription. 4) The patients accompanied with constipation started to take the composition 7. When the constipation was improved after administration, the composition 1 or 4 was taken as the maintenance prescription. The administration time is one month; other indexes were detected with 30-day dosing except the glucose tolerance test was performed once respectively with 14- and 30-day dosing.
Statistical treatment: Pearson correlation coefficient was taken to the correlation, and non-correlation was excluded by paired T test. As for the intergroup control, paired T test was used to compare the change before and after administration (p<0.05 shows significant differences).
<Improvement Effect on Dyslipidemia>
Test Method:
The volunteers were divided into a normal group and an abnormal group according to the Third Report of the National Cholesterol Education Program (NCEP), NIH Publication No. 02-5215. The volunteers received blood examination before administration and with 30-day dosing. The volunteers were forbidden to take any food except drinking water after 21:00 the previous night of the examination. Serum cholesterol was subjected to conventional determination via a cholesterol oxidase method, and triglyceride was subjected to conventional determination via a GPO-PAP enzyme method in a hospital laboratory.
Test indexes of the normal and abnormal blood are shown in the table below respectively:
<Effect on Serum Total Cholesterol>
Table for basic information of the 30 volunteers (16 in the normal group and 14 in the abnormal group) and data change before and after administration
<Effect on Low Density Lipoprotein Cholesterol and High Density Lipoprotein Cholesterol>
Serum total cholesterol is the sum of cholesterol contained in lipoprotein of blood, and mainly consists of low density lipoprotein cholesterol and high density lipoprotein cholesterol. The therapeutic effects of 30 volunteers (12 in the normal low density lipoprotein cholesterol group and 18 in the abnormal low density lipoprotein cholesterol group) were observed in this present invention. Table for basic information of the volunteers and data change before and after administration
The above result indicates that the composition has a good effect of decreasing the low density lipoprotein cholesterol. Low density lipoprotein binds to cholesterol to transport cholesterol into peripheral tissue cells. Too much low density lipoprotein in blood will form residual particles to be accumulated within the blood vessel wall under blood vessel endothelium. This is the critical first step in the formation of coronary atherosclerosis, which breaks the integrity of endothelial barrier, activates proinflammatory cytokines and chemokines, promotes the generation of reactive oxygen (ROS) (Weber, C. et al. Nat. Med. 2011), finally leading to serious diseases endangering life, such as, angina pectoris, heart stroke and cerebral stroke. Therefore, low density lipoprotein cholesterol is generally called harmful cholesterol. The composition of the present invention may significantly decrease the excessive low density lipoprotein cholesterol, and is of important meaning in preventing the occurrence of cardiovascular and cerebrovascular diseases, and their progress.
Currently, the common clinical antilipemic agents mainly include HMG-CoA reductase inhibitors (statins drugs), cation exchange resin preparations, and cholesterol transferase inhibitors of small intestines. These therapeutics not only have the side effects of dissolving striated muscle, impairment of liver and renal functions, but also will affect the physiological functions of cholesterol with excessive amount. The composition consists of medicine-food homologous food, and has a very small possibility of side effect, and may obviously decrease the low density lipoprotein cholesterol (harmful cholesterol), specifically improve hypercholesteremia, free of affecting the physiological level of cholesterol and thus, is of great clinical significance.
<Effect on Triglyceride>
Triglyceride was tested in this present invention. Table for basic information of the 30 volunteers (9 in the normal group and 21 in the abnormal group) and data change before and after administration
<Improvement Effect on Fasting Blood-Glucose>
Test Method
Prediabetes volunteers or volunteers in the early stage of diabetes (fasting blood-glucose (FBG)>5.56 to <7.5 mmol/L) were selected as an abnormal FBG group and volunteers with FBG≤5.56 mmol/L were selected as a normal group based on the standards of American Diabetes Association (ADA). The volunteers received blood examination before administration and with 30-day dosing. The volunteers were forbidden to take any food except drinking water after 21:00 the previous night of the examination. The FBG was determined by a glucose oxidase-peroxidase coupling method in a hospital laboratory. Table for basic information of the 32 volunteers (13 in the normal group and 19 in the abnormal group) and changes of the FBG values before and after administration
The present invention further utilizes glucose tolerance tests to observe the improvement effect of the composition on the insulin resistance of the 8 prediabetes or diabetes volunteers. The volunteers received blood examination before administration and with 14- and 30-day dosing. The volunteers were forbidden to take any food except drinking water after 21:00 the previous night of the examination. In the next morning, venous blood was respectively drawn at 0 min (before taking glucose), 30 min, 1 h and 2 h later after orally taking 75 g glucose solution, thus determining the blood glucose level.
<Improvement Effect on High Uric Acid in Serum>
Test Method
Volunteers with serum uric acid ≥420 mol/L (male) and ≥360 mol/L (female) served as an abnormal serum uric acid group, and volunteers with serum uric acid lower than the level served as a normal serum uric acid group (Liu A D et al, J Atheroscler. Theromb. 2013). The table below shows the basic information of the 28 volunteers (15 in the normal serum uric acid group and 13 in the abnormal serum uric acid group) and changes of the serum uric acid values before and after administration. The volunteers received blood examination before administration and with 30-day dosing. The volunteers were forbidden to take any food except drinking water after 21:00 the previous night of the examination. Serum uric acid was determined by coupled-enzyme assay.
Traditionally, the diseases associated with high-level serum uric acid mainly include symptomatic high-uric acid diseases caused by uric acid crystal, such as gout, acute and chronic arthritis, uric acid-derived kidney stones, acute or chronic renal damage, and the like. In recent years, lots of epidemiological, clinical and experimental studies have indicated that the asymptomatic high-uric acid patients among the crowd have a higher proportion, and the asymptomatic high-uric acid is associated with the pathogenesis of cardiovascular and cerebrovascular diseases, central nervous disorders, diseases of immune and renal systems, erectile dysfunction of disordered sex activity, and sexual apathy and other multiple systemic diseases. The extent of harm on human health caused by these effects of soluble high uric acid is much larger than gout; therefore, high uric acid has been highly concerned in medical and science fields. Hyperuricemia has huge potential risk to human body, but uric acid is an important physiological activator and is of great significance to human health. Uric acid is a kind of powerful ROS antioxidant and peroxynitrite scavenger. Uric acid can scavenge free radicals and chelate metal ions (Frei B, et al., PNAS 1988; Glantzounis G K, et al. Curr Pharm Des 2005; Sautin Y Y, Johnson R J. Nucleosides Nucleotides Nucl Acids 2008). Half of plasma oxidation resistance of human body is from uric acid, and such kind of function is stronger than that of folic acid (Yeum K J, et al., 2004). The physiological level of uric acid may play a protective effect on the integrity of blood vessel endothelium, and is also an important medium to human type-II immunoreaction and plays a crucial role in response to the protective antibody of vaccines (Kool M, et al. J Exp Med 2008; Kool M, et al. Immunity 2011).
The composition may not affect the normal physiological level of uric acid while controlling hyperuricemia, and is of great significance in maintaining the physiological level of uric acid, and preventing or treating diseases associated with high uric acid.
<Improvement Effect on Renal Functions>
Test Method
Serum creatinine and glomerular filtration rate are the basic methods to determine liver functions. The present invention utilized the detection of serum creatinine level and estimated glomerular filtration rate to prove the improvement effect of the composition on renal functions. The volunteers received blood examination before administration and with 30-day dosing. The volunteers were forbidden to take any food except drinking water after 21:00 the previous night of the examination. In the next morning, venous blood was drawn to conventionally determine the serum creatinine level by a creatinine enzyme method. The table below shows the basic information of the 23 volunteers and changes of the serum creatinine level before and after administration.
Effect on the Estimated Glomerular Filtration Rate (eGFR)
Estimation Method
eGFR was calculated by the China's improved approach based on the measured value of the above serum creatinine. Volunteers with eGFR<90 mi/L served as an abnormal glomerular filtration rate group. The table below shows basic information of the volunteers and data change after administration.
<Improvement Effect on Liver Functions>
Test Method
Hepatocellular damage index threshold values obtained by the studies on non-alcoholic fatty liver diseases were used in this present invention (Bedogni G et al. Hepatology 2005). Volunteers with alanine aminotransferase or aspartate aminotransferase >30 U/L served as objects to observe the changes before administration and with 30-day dosing. The volunteers were forbidden to take any food except drinking water after 21:00 the previous night of the examination. In the next morning, venous blood was drawn to determine the activity of alanine aminotransferase or aspartate aminotransferase by a velocity method. The table below shows the basic information of the 10 volunteers and activity changes of the alanine aminotransferase or aspartate aminotransferase before and after administration.
| Number | Date | Country | Kind |
|---|---|---|---|
| 201911239497.1 | Dec 2019 | CN | national |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/CN2020/130277 | 11/20/2020 | WO |
