Sperm-nourishing and liver-soothing composition for treating kidney deficiency, liver stagnation and spleen deficiency type premature ovarian failure and preparation method of essence-nourishing and liver-soothing composition

Abstract

The invention discloses an essence-nourishing and liver-soothing composition for treating kidney deficiency, liver stagnation and spleen deficiency type premature ovarian failure and a preparation method of the essence-nourishing and liver-soothing composition, and belongs to the field of traditional Chinese medicines, the essence-nourishing and liver-soothing composition is prepared from fructus corni, radix rehmanniae preparata, radix angelicae sinensis, radix paeoniae alba, rhizoma dioscoreae, pericarpium citri reticulatae, semen cuscutae, fructus ligustri lucidi, herba lycopi and rhizoma cyperi. The essence-nourishing and liver-soothing soup is based on the essence-nourishing and seed-growing soup, is added and reduced by combining clinical practice and summarized experience, is prepared to tonify kidney and replenish essence, soothe liver, regulate and tonify Chong and Ren meridians, dredge the Ren and dredge and regulate qi and blood, effectively improves the ovarian function, and further achieves the effects of regulating menstruation and assisting pregnancy. The essence-nourishing and liver-soothing soup is mainly used for premature ovarian failure patients, and prophase clinical observation shows that the essence-nourishing and liver-soothing soup has good clinical effects in the aspects of improving serum sex hormone indexes, increasing the number of sinus follicles, promoting follicle maturation and smooth discharge and improving the success rate of pregnancy.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine, specifically relating to a composition for nourishing essence and soothing the liver for treating premature ovarian failure of the kidney deficiency, liver stagnation and spleen deficiency type, and its preparation method. Background Technology

[0002] With the accelerating pace of society, increased psychological stress on women, and the rise of adverse effects from chemotherapy and radiotherapy for cancer, the number of patients with premature ovarian failure (POF) is increasing year by year, and the trend is towards younger women. The main manifestation is hormonal imbalance, leading to hot flashes and ovarian atrophy, ultimately resulting in infertility. Currently, clinical treatment for POF primarily involves estrogen supplementation to alleviate symptoms. However, long-term estrogen use can cause significant side effects, mainly increasing the risk of endometrial and breast cancer. Worse still, the corresponding clinical symptoms reappear immediately after discontinuing medication, even leading to ovarian failure. Currently, the main clinical treatment for infertility caused by POF is assisted reproductive technology (ART). Although ART technology has been continuously optimized, with improvements in ovulation induction, artificial insemination, and embryo culture techniques, the success rate of embryo transfer remains low, and the cost is high. Therefore, it is necessary to explore more effective, safe, and feasible solutions to improve ovarian function and fertility in female patients. Numerous studies have shown that traditional Chinese medicine (TCM) treatment for POF is effective, safe, and inexpensive, and has gradually become a research hotspot in recent years. Traditional Chinese medicine (TCM) utilizes various methods, including syndrome differentiation and treatment, sequential cycle therapy, prepared Chinese medicines, acupuncture, herbal pastes, and retention enemas, to ultimately adjust the reproductive axis of "Kidney-Tian Gui-Chong Ren-Uterus," restoring its balance and thus improving ovarian function. TCM treatment for premature ovarian failure can promote ovarian responsiveness to gonadotropins and increase sex hormone levels, improve ovarian reserve, and promote ovarian function repair. Furthermore, the safety and stability of TCM treatment have been verified, making it of great significance in the prevention and treatment of pathological ovarian dysfunction in women. Summary of the Invention

[0003] The purpose of this invention is to develop a novel combination for treating premature ovarian failure of the kidney-deficiency, liver-stagnation, and spleen-deficiency type, based on the Yangjing Zhongyu Decoction and modified by clinical practice and experience. This combination (Yangjing Shugan Decoction) has the functions of tonifying the kidney and replenishing essence, soothing the liver and strengthening the spleen, regulating the Chong and Ren meridians, making the Ren meridian unobstructed, the Chong meridian full, and the Qi and blood harmonious, effectively improving ovarian function, and thus playing a role in regulating menstruation and assisting fertility.

[0004] The present invention relates to a formula for nourishing essence and soothing the liver, which is composed of 20-30 parts of Cornus officinalis, 10-20 parts of Rehmannia glutinosa, 5-15 parts of Angelica sinensis, 10-20 parts of Paeonia lactiflora, 10-20 parts of Dioscorea opposita, 5-15 parts of Citrus reticulata, 10-20 parts of Cuscuta chinensis, 10-20 parts of Ligustrum lucidum, 5-15 parts of Lycopus lucidus, and 10-20 parts of Cyperus rotundus.

[0005] Furthermore, the aforementioned Nourishing Essence and Soothing Liver Decoction is composed of 25-30 parts of Cornus officinalis, 15-20 parts of Rehmannia glutinosa, 10-15 parts of Angelica sinensis, 15-20 parts of Paeonia lactiflora, 15-20 parts of Dioscorea opposita, 10-15 parts of Citrus reticulata, 15-20 parts of Cuscuta chinensis, 15-20 parts of Ligustrum lucidum, 10-15 parts of Lycopus lucidus, and 15-20 parts of Cyperus rotundus, according to the weight proportions.

[0006] Furthermore, the aforementioned Nourishing Essence and Soothing Liver Decoction is composed of 25 parts Cornus officinalis, 16 parts Rehmannia glutinosa, 10 parts Angelica sinensis, 15 parts Paeonia lactiflora, 15 parts Dioscorea opposita, 10 parts Citrus reticulata, 15 parts Cuscuta chinensis, 15 parts Ligustrum lucidum, 10 parts Lycopus lucidus, and 15 parts Cyperus rotundus, according to the weight proportions.

[0007] Furthermore, the aforementioned Nourishing Essence and Soothing Liver Decoction is composed of 30 parts Cornus officinalis, 20 parts Rehmannia glutinosa, 15 parts Angelica sinensis, 15 parts Paeonia lactiflora, 15 parts Dioscorea opposita, 10 parts Citrus reticulata, 15 parts Cuscuta chinensis, 15 parts Ligustrum lucidum, 10 parts Lycopus lucidus, and 15 parts Cyperus rotundus, according to the weight proportions.

[0008] The method for preparing the essence-nourishing and liver-soothing decoction of the present invention is as follows:

[0009] Weigh out 20-30 parts of Cornus officinalis, 10-20 parts of Rehmannia glutinosa, 5-15 parts of Angelica sinensis, 10-20 parts of Paeonia lactiflora, 10-20 parts of Dioscorea opposita, 5-15 parts of Citrus reticulata, 10-20 parts of Cuscuta chinensis, 10-20 parts of Ligustrum lucidum, 5-15 parts of Lycopus lucidus, and 10-20 parts of Cyperus rotundus according to the following proportions: mix them together, add 8-10 times the amount of water, soak for half an hour, decoct for half an hour, filter out the liquid, add 10 times the amount of water, decoct for half an hour, filter out the liquid, repeat twice; combine the three liquids, concentrate, dry, and pulverize to obtain a dry extract powder.

[0010] Explanation of the formula for the Nourishing Essence and Soothing Liver Decoction of this invention:

[0011] Cornus officinalis: It tastes sour and astringent, and is slightly warm. It enters the liver and kidney meridians. It can both nourish the kidneys and replenish essence, and warm the kidneys and assist yang. It is an essential medicine for nourishing the liver and kidneys.

[0012] Rehmannia glutinosa: It is sweet in taste, slightly warm in nature, and moist in texture. It enters the liver and kidney meridians. It can nourish blood and yin, as well as replenish essence and marrow. It is an essential medicine for nourishing the liver and kidneys.

[0013] Angelica sinensis: It has a sweet and pungent taste, is warm in nature, and enters the liver, heart, and spleen meridians. Its functions are to nourish blood, promote blood circulation, regulate menstruation, and relieve pain.

[0014] White peony root: It tastes bitter, sour and sweet, and is slightly cold. It nourishes blood and regulates menstruation, nourishes blood and softens the liver, relieves pain, and astringes yin and stops sweating.

[0015] Yam: Sweet, neutral. It enters the spleen, lung, and kidney meridians. It tonifies the spleen and stomach, generates fluids and nourishes the lungs, and tonifies the kidneys and replenishes essence.

[0016] Dried tangerine peel: pungent and bitter in taste, warm in nature, and enters the spleen and lung meridians. It has the functions of regulating qi and strengthening the spleen, drying dampness and resolving phlegm. It is an excellent medicine for regulating qi and strengthening the spleen.

[0017] Cuscuta seed: It is sweet in taste and warm in nature. It nourishes the liver and kidneys, strengthens essence, and calms the fetus. It not only tonifies kidney yang but also assists kidney yin, and also nourishes the liver, warms the spleen, and aids the stomach.

[0018] Privet fruit: sweet and bitter, cool in nature, nourishes the liver and kidneys, and clears deficiency heat.

[0019] Cyperus rotundus: pungent, slightly bitter, slightly sweet, neutral; enters the liver, spleen, and triple burner meridians; soothes the liver and regulates qi, regulates menstruation and relieves pain; it is a qi-regulating herb in the blood.

[0020] Ze Lan: This herb is bitter and pungent in taste, slightly warm in nature, and enters the liver and spleen meridians. It has the functions of promoting blood circulation, removing blood stasis, regulating menstruation, and promoting diuresis and reducing swelling.

[0021] The formula uses Rehmannia glutinosa, Cornus officinalis, Dioscorea opposita, Cuscuta chinensis, and Ligustrum lucidum as the principal herbs to nourish the liver, spleen, and kidneys, and replenish blood and essence. Angelica sinensis, Paeonia lactiflora, Citrus reticulata peel, and Cyperus rotundus are the assistant herbs, working together to tonify the kidneys, soothe the liver, strengthen the spleen, nourish blood, and promote circulation without causing stagnation. Lycopus lucidus is used as an adjuvant herb to invigorate blood and regulate menstruation. These adjuvant herbs, based on the kidney-tonifying, liver-soothing, and spleen-strengthening properties of the principal and assistant herbs, invigorate blood and regulate the Chong and Ren meridians, thus regulating menstruation. This ensures that the entire formula tonifies while promoting circulation, nourishes without being cloying, and invigorates blood without harming the body.

[0022] The formula is meticulously formulated, addressing both the root cause and symptoms, nourishing without causing stagnation, and replenishing both essence and blood. It also harmonizes the liver, spleen, and kidneys, while simultaneously regulating qi, resolving phlegm, and removing blood stasis. This restores the balance and harmony of "kidney qi, yin and yang, spleen and uterus," resulting in regular menstruation and enabling conception.

[0023] Basic Formula: Nourishing Essence and Fertility Decoction, Source: *Fu Qingzhu's Gynecology* (Qing Dynasty). Composition: Cornus officinalis, Rehmannia glutinosa, Angelica sinensis, Paeonia lactiflora. Core Pathogenesis: Liver and kidney yin deficiency, insufficient essence and blood. Treatment Principle: Nourish the kidneys and blood, regulate and replenish the Chong and Ren meridians. Formula Explanation: Rehmannia glutinosa and Cornus officinalis: Greatly replenish kidney yin, fill essence and benefit marrow, serving as the principal herbs. Angelica sinensis and Paeonia lactiflora: Nourish blood and soften the liver, regulate and replenish the Chong and Ren meridians, serving as the assistant herbs. The four herbs work together, focusing on "nourishing essence" and "fertility" (i.e., assisting conception), regulating female reproductive function by nourishing water (kidney) and nourishing wood (liver). Characteristics: Simple and potent, but tends to be nourishing and stagnant. Targets liver and kidney yin deficiency syndrome of "pure deficiency without pathogenic factors," but if the patient also has qi stagnation, blood stasis, or spleen deficiency with dampness, using this formula alone may hinder digestion, leading to abdominal distension, poor appetite, and other problems. "Nourishing Essence and Soothing Liver Decoction" can be seen as an optimized version of "Nourishing Essence and Fertility Decoction." Building upon the original formula's core principle of "nourishing essence," this version incorporates multiple functionalities, making its clinical application broader, more precise, and more effective. The technological advantages are specifically reflected in the following aspects:

[0024] 1. Harmonizing the liver, spleen, and kidneys in unison deepens the understanding of the subject.

[0025] Foundation (Kidney and Liver): Retains all the core ingredients of the "Nourishing Essence and Cultivating Jade Decoction" (Cornus officinalis, Rehmannia glutinosa, Angelica sinensis, and Paeonia lactiflora) to ensure the fundamental function of nourishing the kidneys and liver and replenishing essence and blood remains unchanged. Strengthening (Spleen): Added Dioscorea opposita and Citrus reticulata peel. Dioscorea opposita: Strengthens the spleen and replenishes qi, consolidates the kidneys and benefits essence. It not only aids spleen function and prevents the cloying nature of Rehmannia glutinosa from hindering stomach function, but also assists in tonifying the kidneys, embodying the idea of ​​"nurturing the innate through the acquired." Citrus reticulata peel: Regulates qi and strengthens the spleen, dries dampness and harmonizes the stomach. It is the finishing touch in the formula, promoting the absorption and transformation of all the ingredients, replenishing without stagnation.

[0026] Compared with the "Nourishing Essence and Soothing Liver Decoction", the present invention improves the treatment from simply treating the liver and kidneys to regulating the liver, spleen and kidneys, resulting in a richer treatment level and better meeting the complex physical conditions of most patients in clinical practice.

[0027] 2. Combining "supplementation" and "unblocking" creates an even more ingenious structure.

[0028] "Tonifying": Rehmannia glutinosa, Cornus officinalis, Angelica sinensis, Paeonia lactiflora, Dioscorea opposita, Cuscuta chinensis, and Ligustrum lucidum are all tonifying herbs. "Unblocking": Cyperus rotundus, Lycopus lucidus, and Citrus reticulata are added. Stir-fried Cyperus rotundus: Revered as the "general manager of qi-related diseases and the commander-in-chief of gynecology," it can soothe the liver, regulate qi, regulate menstruation, and relieve pain. It directly addresses a major factor contributing to menstrual irregularities and infertility caused by liver qi stagnation. Lycopus lucidus: Promotes blood circulation, removes blood stasis, and reduces swelling. Adding a blood-activating herb to a large amount of tonifying herbs prevents stagnation, promotes blood circulation, and improves the pelvic environment, which is especially crucial for patients with blood stasis tendencies (such as dysmenorrhea, blood clots, and petechiae on the tongue).

[0029] Compared to the "Nourishing Essence and Soothing Liver Decoction," the present invention's Nourishing Essence and Soothing Liver Decoction changes the original formula's structure of "only replenishing without clearing," achieving "replenishment with circulation, stillness with movement." It can not only replenish deficient essence and blood but also clear stagnant qi and blood, which is more conducive to nourishing the uterus and its vessels and restoring their function.

[0030] 3. It nourishes kidney yin and warms kidney yang simultaneously, thus replenishing both yin and yang.

[0031] Nourishing Kidney Yin: Rehmannia glutinosa, Cornus officinalis, and Ligustrum lucidum are all excellent remedies for nourishing kidney yin. Warming Kidney Yang: Cuscuta chinensis is added. Cuscuta chinensis tonifies the kidneys, strengthens essence, nourishes the liver, and improves eyesight. It is neutral to slightly warm in nature, capable of both assisting yang and benefiting yin. Its addition allows the entire formula to nourish kidney yin while subtly generating kidney yang, conforming to the principle of "seeking yang within yin." Only when yin and yang are balanced in the kidneys can the uterus be better warmed, promoting ovulation and conception.

[0032] Compared with "Nourishing Essence and Soothing Liver Decoction", the present invention avoids the drawbacks of cold stagnation that may result from simply nourishing Yin, making the kidney-tonifying power more balanced and comprehensive, and closer to the state of "abundant kidney qi" required for reproductive function.

[0033] 4. Precisely targets modern causes of disease, making it more widely applicable.

[0034] Modern women face high levels of stress, which can easily lead to liver qi stagnation; prolonged sitting and lack of exercise can easily lead to poor blood circulation; and irregular eating habits can easily lead to spleen deficiency and dampness accumulation. The formulation of this invention's "Nourishing Essence and Soothing Liver Decoction" precisely addresses these common modern pathogenesis mechanisms:

[0035] Liver Qi stagnation: Cyperus rotundus and dried tangerine peel soothe the liver and regulate Qi. Blood stasis: Lycopus lucidus and Angelica sinensis invigorate blood circulation. Spleen deficiency: Dioscorea opposita and dried tangerine peel strengthen the spleen. Kidney deficiency (Yin and Yang deficiency): Rehmannia glutinosa, Cornus officinalis, Cuscuta chinensis, etc., tonify the kidneys.

[0036] Therefore, for menstrual disorders, premenstrual breast distension, dysmenorrhea, and infertility caused by complex syndromes such as kidney essence deficiency, liver stagnation, blood stasis, and spleen weakness, the therapeutic effect will be more significant than that of "Yangjing Zhongyu Tang" alone.

[0037] in conclusion:

[0038] This invention, "Nourishing Essence and Soothing Liver Decoction," is not a simple additive formula, but a hierarchical optimized formula. While firmly grasping the fundamental principle of "nourishing essence," it greatly enhances the original formula's clinical relevance, applicability, and therapeutic effectiveness by adding herbs that regulate qi, invigorate blood, and strengthen the spleen. It achieves an upgrade from "single-point breakthrough" to "systemic conditioning," serving as a model for the modern application of an ancient formula.

[0039] Raw materials and their functions:

[0040] Eupatorium fortunei: Bitter, pungent, slightly warm. Enters the liver and spleen meridians. Invigorates blood circulation and regulates menstruation, removes blood stasis and reduces swelling, promotes diuresis and reduces edema. Used for irregular menstruation, amenorrhea, dysmenorrhea, postpartum abdominal pain due to blood stasis, carbuncles and boils, edema and ascites. *Benjing Fengyuan* states: Eupatorium fortunei specifically treats postpartum blood stasis, flowing to the waist and thighs, causing cramps and pain, breaks up stagnant blood, eliminates masses, removes edema, and treats swelling of the body, face, and limbs. *Benjing* also states that it treats traumatic injuries, carbuncles, and boils, all due to its blood-dispersing effect, making it a key herb in obstetrics. When combined with Ligusticum chuanxiong, Angelica sinensis, and child's urine, its efficacy surpasses that of Leonurus japonicus.

[0041] Cornus officinalis: Sour, astringent, slightly warm. It enters the liver and kidney meridians. It tonifies the liver and kidneys, and astringes and consolidates the body. It is used for dizziness and tinnitus, lower back and knee pain, impotence and seminal emission, enuresis and frequent urination, metrorrhagia and leukorrhea, profuse sweating and collapse, and internal heat and thirst.

[0042] Rehmannia glutinosa (processed): Sweet, slightly warm. Enters the liver and kidney meridians. Nourishes blood and yin, replenishes essence and marrow. Used for blood deficiency and chlorosis, palpitations, irregular menstruation, metrorrhagia, liver and kidney yin deficiency, soreness and weakness of the lower back and knees, bone steaming fever, night sweats, seminal emission, internal heat and thirst, dizziness, tinnitus, and premature graying of hair. *Ben Cao Cong Xin* states: Nourishes kidney water, fills bone marrow, benefits blood vessels, replenishes true yin, improves hearing and vision, and blackens hair and beard. It also replenishes edema, treats chronic diarrhea, treats fatigue, wind-dampness, fever due to yin deficiency, dry cough with phlegm, shortness of breath, hunger in the stomach, smallpox with heart deficiency and no pus, post-illness leg and thigh pain, postpartum abdominal pain, yin deficiency due to pathogenic factors, anhidrosis, and various other ailments. It is the main medicine for strengthening yin.

[0043] Angelica sinensis: Sweet, pungent, and warm. It enters the liver, heart, and spleen meridians. It nourishes and invigorates blood, regulates menstruation and relieves pain, and moistens the intestines to relieve constipation. It is used for blood deficiency and chlorosis, dizziness and palpitations, irregular menstruation, amenorrhea and dysmenorrhea, abdominal pain due to deficiency and cold, rheumatic arthralgia, injuries from falls, carbuncles and sores, and constipation due to intestinal dryness. Angelica sinensis soaked in wine invigorates blood and regulates menstruation. It is used for amenorrhea and dysmenorrhea, rheumatic arthralgia, and injuries from falls. The *Annotations on the Treatise on Cold Damage* states: "The pulse is the residence of blood, and all blood belongs to the heart. Therefore, to unblock the pulse, one must first nourish the heart and replenish blood. Thus, Zhang Zhongjing treated cold extremities and a weak, almost imperceptible pulse by using the bitter and warm nature of Angelica sinensis to assist the heart blood." The *Secret Formulas for Treatment* states: "Angelica sinensis has three uses: first, it is a primary medicine for the heart meridian; second, it harmonizes blood; and third, it treats various diseases that worsen at night." For treating external ailments, it must be soaked in wine to break down hardened masses. It is essential for all diseases affecting the blood. For unbearable eye pain, a decoction of Coptis chinensis and Angelica sinensis root soaked in wine is taken orally. It is also said that when blood is congested and does not flow, there is pain. Angelica sinensis, with its pungent and warm properties, can disperse the blood and qi, allowing them to flow freely.

[0044] White peony root: Bitter, sour, slightly cold. It enters the liver and spleen meridians. It nourishes blood and regulates menstruation, astringes yin and stops sweating, softens the liver and relieves pain, and calms liver yang. It is used for blood deficiency and chlorosis, irregular menstruation, spontaneous sweating, night sweats, hypochondriac pain, abdominal pain, limb spasms, headache, and dizziness. The *Shennong Bencao Jing* states: "It treats abdominal pain caused by pathogenic factors, eliminates blood stasis, breaks up hard masses, treats hernia due to cold and heat, relieves pain, promotes urination, and benefits qi." The *Yaoxing Lun* states: "It treats lung pathogenic factors, abdominal pain, blood stasis, clears stagnation in the internal organs, treats pathogenic pain and blood stasis, treats seasonal diseases and bone fever, strengthens the five internal organs, tonifies kidney qi, treats abdominal distension, amenorrhea in women, eliminates blood stasis, and can erode pus."

[0045] Yam: Sweet, neutral. Enters the spleen, lung, and kidney meridians. Tonifies the spleen and stomach, generates fluids and benefits the lungs, tonifies the kidneys and astringes essence. Used for spleen deficiency with poor appetite, chronic diarrhea, lung deficiency with cough and asthma, kidney deficiency with seminal emission, leukorrhea, frequent urination, and thirst due to deficiency heat. Wheat bran-fried yam tonifies the spleen and stomach. Used for spleen deficiency with poor appetite, loose stools, and excessive leukorrhea. *Shennong's Classic of Materia Medica*: "Treats internal injuries, tonifies deficiency, eliminates cold and heat pathogens, tonifies the middle jiao and strengthens qi, builds muscle, and with prolonged use, improves hearing and vision." *Supplement to Shennong's Classic of Materia Medica*: "Treats facial and head wind, dizziness, lowers qi, stops lower back pain, treats weakness and emaciation, nourishes the five internal organs, eliminates vexation and heat, and strengthens the body."

[0046] Dried tangerine peel: bitter, pungent, and warm. It enters the lung and spleen meridians. It regulates qi and strengthens the spleen, dries dampness and resolves phlegm. It is used for abdominal distension, poor appetite, vomiting and diarrhea, and cough with excessive phlegm.

[0047] Cuscuta seed: pungent, sweet, neutral. Enters the liver, kidney, and spleen meridians. Tonifies the liver and kidneys, strengthens essence and reduces urination, calms the fetus, improves eyesight, and stops diarrhea; externally used to dispel wind and remove blemishes. Used for liver and kidney deficiency, lower back and knee weakness, impotence, seminal emission, enuresis, frequent urination, kidney deficiency with threatened abortion, threatened miscarriage, blurred vision and tinnitus, spleen and kidney deficiency with diarrhea; externally used to treat vitiligo. First recorded in the *Shennong Bencao Jing* (Shennong's Classic of Materia Medica), it was listed as a superior herb. Sweetness generally has a nourishing effect; sweet, pungent, and slightly warm, with a neutral nature, it can tonify both yang and yin, possessing the characteristics of being warm without being drying and tonifying without causing stagnation. Its effects on strengthening essence and calming the fetus are similar to those of sex hormones. Traditional Chinese medicine believes that the kidney is the root of innate essence, containing both primordial yin and primordial yang, the source of human reproduction and development, and the driving force of organ function.

[0048] Cyperus rotundus: pungent, slightly bitter, slightly sweet, neutral. It enters the liver, spleen, and triple burner meridians. It soothes the liver and relieves stagnation, regulates qi and relieves chest tightness, regulates menstruation and relieves pain. It is used for liver qi stagnation, chest and rib pain, hernia pain, breast pain, spleen and stomach qi stagnation, abdominal distension and pain, irregular menstruation, amenorrhea, and dysmenorrhea.

[0049] The present invention has the following beneficial effects:

[0050] This invention theoretically inherits the academic thought of Yang Zongmeng, clearly proposing that the core pathogenesis of premature ovarian failure is "kidney deficiency as the root cause and liver stagnation as the manifestation," thus establishing an innovative approach to treatment based on "kidney deficiency and liver stagnation." Guided by this theory, and based on the "Nourishing Essence and Cultivating Jade Decoction" from *Fu Qingzhu's Gynecology*, a self-formulated "Nourishing Essence and Soothing Liver Decoction" is developed for holistic treatment, using the principles of "tonifying the kidneys and replenishing essence, soothing the liver and strengthening the spleen, and regulating the Chong and Ren meridians." This effectively improves ovarian function, thereby regulating menstruation and aiding fertility.

[0051] The Yangjing Shugan Decoction is mainly for patients with premature ovarian failure. Preliminary clinical observations have shown that the Yangjing Shugan Decoction has good clinical effects in improving serum sex hormone levels, increasing the number of antral follicles, promoting follicle development and maturation, facilitating ovulation, and improving the success rate of pregnancy. Attached Figure Description

[0052] Figure 1The LC-MS / MS total ion chromatograms of YJSGD are shown in the following modes: (A) negative ion mode; (B) positive ion mode. In the chromatograms, 1: rehmannia glycoside D; 2: monoglucoside; 3: loganin; 4: paeoniflorin; 5: ferulic acid; 6: hyperoside; 7: privetin; 8: hesperidin; 9: luteolin; 10: ursolic acid.

[0053] Figure 2 This is a graph showing the changes in mouse body weight.

[0054] Figure 3 Comparison of serum E2, FSH and LH levels in mice of different groups (Mean±SD, n=10). ,

[0055] vs. blank group; , vs. model group;

[0056] Figure 4 The effect of Yangjing Shugan Decoction on the serum levels of SOD, CAT, GSH-Px and MDA in mice with oxidative damage (Mean±SD, n=10). , vs. blank group; , vs. model group;

[0057] Figure 5 Figure showing the effect of Yangjing Shugan Decoction on serum IL-1β, IL-6 and TNF-α levels in mice with oxidative damage; (Mean±SD, n=10) , vs. blank group; , vs. model group;

[0058] Figure 6 Figure 1: Effect of Yangjing Shugan Decoction on the gut microbiota of POF mice; A: Dilution curve; B: Shannon curve; CF: Alpha diversity analysis results (C: ACE; D: Chao1; E: Simpson; F: Shannon); GH: Beta diversity analysis (G: PCA; H: PCoA).

[0059] Figure 7 The effect of Yangjing Shugan Decoction on the phylum-level intestinal flora of POF mice; A: Phylum-level bar chart; BC: Comparison of characteristic flora;

[0060] Figure 8 Effects of Yangjing Shugan Decoction on the genus-level intestinal flora of POF mice; A: Genus-level bar chart; BE: Comparison of characteristic flora; F: Genus-level heatmap;

[0061] Figure 9 Figure 1 shows the serum metabolomics analysis (n=6); (A) OPLS-DA analysis of samples under positive ion mode; (B) OPLS-DA analysis of control group and model group; (C) OPLS-DA analysis of model group and YJSGD-M group; (D) OPLS-DA analysis of samples under negative ion mode; (E) OPLS-DA analysis of control group and model group; (F) OPLS-DA analysis of model group and YJSGD-M group.

[0062] Figure 10 Metabolomics analysis of Yangjing Shugan Decoction in POF mice; A: Cluster analysis of differentially expressed metabolites; BC: Metabolic pathway analysis of differentially expressed metabolites. Detailed Implementation

[0063] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the spirit of the contents disclosed in the present invention will be described in detail below. After understanding the embodiments of the present invention, any person skilled in the art can make changes and modifications based on the technology taught in the present invention without departing from the spirit and scope of the present invention.

[0064] The illustrative embodiments and descriptions of the present invention are used to explain the present invention, but are not intended to limit the present invention.

[0065] Example 1

[0066] The essence-nourishing and liver-soothing decoction in this embodiment is made from 20g of Cornus officinalis, 15g of Rehmannia glutinosa, 10g of Angelica sinensis, 15g of Paeonia lactiflora, 15g of Dioscorea opposita, 15g of Citrus reticulata peel, 15g of Cuscuta chinensis, 15g of Ligustrum lucidum, 10g of Lycopus lucidus, and 15g of Cyperus rotundus.

[0067] 1. Preparation process of Nourishing Essence and Soothing Liver Decoction

[0068] Take 20g of Cornus officinalis, 15g of Rehmannia glutinosa, 10g of Angelica sinensis, 15g of Paeonia lactiflora, 15g of Dioscorea opposita, 15g of Citrus reticulata peel, 15g of Cuscuta chinensis, 15g of Ligustrum lucidum, 10g of Lycopus lucidus, and 15g of Cyperus rotundus. Add 8-10 times the amount of water, soak for half an hour, decoct for half an hour, filter the liquid, add 10 times the amount of water, decoct for half an hour, filter the liquid, and repeat twice. Combine the three liquids, concentrate, dry, and pulverize to obtain a dry extract powder (64.28g), with an extract yield of 44.32%.

[0069] Example 2

[0070] The drug for treating premature ovarian failure prepared in the above embodiments was applied clinically. A total of 18 patients were included in the study. All patients visited the outpatient clinics of the Affiliated Hospital and the Third Affiliated Clinical Hospital of Changchun University of Traditional Chinese Medicine from January 2021 to June 2025 and met the diagnostic criteria for delayed menstruation and infertility caused by premature ovarian failure of the liver stagnation, kidney deficiency and spleen deficiency type.

[0071] Traditional Chinese Medicine (TCM) diagnostic criteria: Referring to the relevant chapters on "scanty menstruation, delayed menstruation, and amenorrhea" in the "Guidelines for Diagnosis and Treatment of Common Gynecological Diseases in Traditional Chinese Medicine" (2012) and "Traditional Chinese Medicine Gynecology", the criteria for diagnosing premature ovarian failure as "kidney deficiency, liver stagnation, and spleen deficiency syndrome" were established, and the diagnosis and scoring were carried out in combination with the main symptoms, secondary symptoms, and tongue and pulse.

[0072] (1) Main symptoms: delayed menstrual cycle or even amenorrhea, and scanty menstrual flow.

[0073] (2) Secondary symptoms: hot flashes and night sweats, soreness and weakness of the waist and knees, insomnia and dreaminess, dizziness and tinnitus, fatigue, vaginal dryness, irritability and anger, dry mouth and bitter taste, and irregular bowel movements.

[0074] (3) Tongue and pulse: The tongue is red or dark red with little coating, teeth marks on the sides, and the pulse is thin and hesitant.

[0075] All patients were treated with a decoction for nourishing essence and soothing the liver, one dose per day. The decoction was prepared in 300mL portions, divided into two warm doses of 150mL each, taken twice daily (morning and evening). One course of treatment lasted 28 days, with three consecutive courses administered. Serum hormone levels (FSH, LH, E2, AMH) and ovarian reserve indicators (OV, AFC) before and after treatment were used as the primary evaluation indicators.

[0076] Before and after treatment, the Yangjing Shugan Decoction significantly reduced FSH and LH levels, increased E2 levels, helped reverse the decline in ovarian function, promoted follicle development and menstrual recovery, and enhanced natural pregnancy. It improved related symptoms and laboratory indicators in a short period, and long-term follow-up showed a relatively lasting effect and stable overall efficacy. Typical cases are as follows:

[0077] Case 1: Ms. Zhang, 36 years old. Initial visit: May 6, 2018.

[0078] Chief complaint: Amenorrhea for 4 months, accompanied by paroxysmal hot flashes and sweating, and irritability for 1 month.

[0079] Present Illness: The patient had her first menstruation at age 13. Her previous menstrual cycles were regular, with a cycle of 5-7 days / 30-35 days, moderate flow, bright red color, no blood clots, and no dysmenorrhea. LMP: Absent since January 6, 2018. For the past month, she has experienced paroxysmal hot flashes and sweating, accompanied by irritability; significantly reduced vaginal discharge, fatigue, good appetite, poor sleep with frequent awakenings, normal urination, and loose stools 1-2 times / day. Physical Examination: Red tongue with a thin, yellow coating and teeth marks on the edges; deep, thready, weak, and slightly hesitant pulse.

[0080] Obstetric history: G2P1, 1 cesarean section, 1 induced abortion, currently using barrier contraception.

[0081] Physical and chemical examination (May 6, 2018): Gynecological examination: No obvious abnormalities. AMH: 0.19 ng / mL; Sex hormones: FSH: 45.56 mIU / mL, LH: 38.47 mIU / mL, E2: 73.9 pg / mL. Vaginal ultrasound: Uterus anteverted, size: 3.7cm × 4.1cm × 3.5cm, endometrial thickness 0.4cm; right ovary size: 1.8cm × 1.6cm, no obvious follicles seen; left ovary size: 2.3cm × 1.5cm, one follicle seen, size 0.6cm × 0.5cm.

[0082] Western medical diagnosis: premature ovarian failure.

[0083] Traditional Chinese Medicine diagnosis: Delayed menstruation (Kidney deficiency, liver stagnation, and spleen deficiency syndrome).

[0084] Treatment: Tonify the kidneys and replenish essence, soothe the liver and strengthen the spleen, and regulate the Chong and Ren meridians.

[0085] Prescription: Nourishing Essence and Soothing Liver Decoction. One dose of the above Chinese herbs daily. Decoct to obtain 300mL of liquid, divided into two warm doses, 150mL each time, morning and evening. One course of treatment is 28 days, with three consecutive courses. After the first course (June 8, 2018), the patient's hot flashes and sweating symptoms significantly decreased, occasional irritability and fatigue were relieved, appetite improved, sleep improved, and bowel movements were normal. Follow-up sex hormone tests showed: FSH: 26.7 mIU / mL, LH: 19.09 mIU / mL, E2: 29.99 pg / mL, indicating a reversal trend in ovarian function. After the second course (July 10, 2018), menstruation occurred during treatment (July 9, 2018), with light flow, dark red color, occasional hot flashes, no sweating, occasional fatigue, significantly increased vaginal discharge, appetite improved, sleep significantly improved, and bowel movements were normal. Follow-up sex hormone tests: FSH: 15.05 mIU / mL, LH: 10.46 mIU / mL, E2: 14.6 pg / mL. Follow-up vaginal ultrasound: Uterus anteverted, measuring 4.1cm × 4.3cm × 3.5cm, En 0.4cm; Right ovary: 1.8cm × 1.8cm, containing one follicle measuring 0.6cm × 0.3cm; Left ovary: 2.3cm × 1.5cm, containing two follicles measuring 0.8cm × 0.5cm and 0.6cm × 0.5cm respectively. After the third course of treatment (August 20, 2018), LMP: Menstruation began on August 19, 2018, with slightly increased flow, dark red color, no hot flashes, no sweating, moderate discharge, good appetite, good sleep, and normal bowel movements. Follow-up sex hormone tests: FSH: 12.05 mIU / mL, LH: 9.06 mIU / mL, E2: 30.6 pg / mL. Follow-up transvaginal ultrasound: Uterus anteverted, measuring 4.3cm × 4.3cm × 3.5cm, En 0.67cm; Right ovary: 2.1cm × 1.8cm, containing 2 follicles, the larger one measuring 0.6cm × 0.3cm; Left ovary: 2.3cm × 1.5cm, containing 2 follicles, the larger one measuring 0.6cm × 0.6cm. Six-month follow-up showed regular menstruation, vaginal lubrication, significantly increased discharge, and improved mood.

[0086] Case 2: Zhao, female, 31 years old. Initial visit: September 4, 2022.

[0087] Chief complaint: Infertility after 5 years of marriage, menstrual cycle is 1 year late.

[0088] Present Illness: The patient had her first menstruation at age 12. She reported irregular menstrual cycles of 3-7 / 40-60 days for the past year, with scanty flow, bright red color, no blood clots, and no dysmenorrhea. Last menstrual period: August 22, 2022, M14. Current symptoms: Occasional hot flashes and sweating, accompanied by irritability; very little vaginal discharge, no lower back or abdominal pain, good appetite, poor sleep, easily awakened, normal urination, and stools that are sometimes dry and sometimes loose. Physical Examination: Pale and swollen tongue with a thin, slightly dark coating and teeth marks on the edges; deep, thready, and slightly hesitant pulse.

[0089] Obstetric history: Married for 5 years, normal sexual activity, G0, no pregnancy for 3 years without contraception.

[0090] Physical and chemical examinations (September 4, 2022): Gynecological examination: No abnormalities. AMH: 0.69 ng / mL; Sex hormone tests: FSH: 40.89 mIU / mL, LH: 35.41 mIU / mL, E2: 55.9 pg / mL. Vaginal ultrasound: Uterus anteverted, size: 3.5cm × 4.3cm × 3.4cm, endometrial thickness 0.4cm, left ovary 2.3cm × 1.5cm, containing 2 follicles, the larger one 0.4cm × 0.4cm; right ovary 1.8cm × 1.6cm, no follicles seen.

[0091] Western medical diagnosis: premature ovarian failure.

[0092] Traditional Chinese Medicine diagnosis: Delayed menstruation, infertility (Kidney deficiency, liver stagnation, and spleen deficiency syndrome).

[0093] Treatment: Tonify the kidneys and replenish essence, soothe the liver and strengthen the spleen, and regulate the Chong and Ren meridians.

[0094] Prescription: Nourishing Essence and Soothing Liver Decoction. One dose of the above Chinese herbs daily. Decoct to obtain 300mL of liquid, divided into two warm doses, 150mL each time, morning and evening. One course of treatment is 28 days, with three consecutive courses. After the first course (September 22, 2022), the patient's hot flashes and sweating symptoms significantly decreased, occasional irritability improved, sleep improved, urination was normal, and bowel movements were sometimes dry and sometimes loose. Follow-up sex hormone tests showed: FSH: 29.04 mIU / mL, LH: 22.70 mIU / mL, E2: 76.47 pg / mL, indicating initial improvement in ovarian function. After the second course (October 28, 2022), menstruation began on October 25, 2022, with very light flow, brownish color, and lasting only two days. Sleep significantly improved, and bowel movements became regular. Follow-up sex hormone tests: FSH: 17.06 mIU / mL, LH: 10.56 mIU / mL, E2: 53.2 pg / mL. Follow-up vaginal ultrasound: Uterus anteverted, measuring 3.8cm × 4.3cm × 3.6cm, En 0.6cm; Right ovary: 2.1cm × 1.6cm, containing two very small follicles measuring 0.3cm × 0.2cm and 0.4cm × 0.3cm; Left ovary: 2.3cm × 1.5cm, containing two follicles measuring 0.7cm × 0.5cm and 0.6cm × 0.5cm. After the third course of treatment: Menstrual cycle returned to 30-33 days, with moderate flow, bright red color, and regular menstruation. Appetite and sleep were good, and bowel movements were normal. After the fifth course of treatment (January 31, 2023), the patient had amenorrhea for 41 days (LMP: December 22, 2022), accompanied by breast tenderness. A blood test for β-HCG showed a level of 122 mIU / mL, confirming pregnancy. The pregnancy proceeded smoothly, culminating in the delivery of a healthy baby boy at full term. Both mother and child are safe and healthy.

[0095] Case 3: Ms. Sun, 38 years old. Initial visit: October 31, 2023.

[0096] Chief complaint: Reduced menstrual flow for 3 years, amenorrhea for 5 months.

[0097] Present Illness: The patient had her first menstruation at age 14. Her previous menstrual cycle was 5 / 28–60 days. In the past 3 years, her menstrual flow has significantly decreased by 2 / 3, becoming dark red with occasional blood clots. She has no dysmenorrhea. Last Menstrual Period: No menstruation since May 14, 2023. Current Symptoms: Paroxysmal hot flashes, significant sweating, accompanied by irritability, vaginal dryness, good appetite, poor sleep with frequent awakenings, normal urination, and loose stools 1-2 times / day. Physical Examination: Red tongue with petechiae on the edges, thin yellow coating with teeth marks on the edges, and a deep, thready, slightly hesitant pulse.

[0098] Obstetric history: Married, G1P1, gave birth to one girl vaginally, currently using condoms for contraception.

[0099] Physical and chemical examination (October 16, 2023): Gynecological examination: No obvious abnormalities. Sex hormone test: FSH: 75.56 mIU / mL, LH: 45.47 mIU / mL, E2: 30.7 pmol / L. Vaginal ultrasound: Uterus anteverted, size: 3.6cm × 4.0cm × 3.7cm, endometrial thickness: 0.3cm; right ovary, size: 2.0cm × 1.2cm, containing one follicle, size: 0.4cm × 0.5cm; left ovary, size: 1.8cm × 1.3cm, no obvious follicle seen, echogenicity is solid.

[0100] Western medical diagnosis: premature ovarian failure.

[0101] Traditional Chinese Medicine diagnosis: Amenorrhea (Kidney deficiency, liver stagnation, and spleen deficiency syndrome).

[0102] Treatment: Tonify the kidneys and replenish essence, soothe the liver and strengthen the spleen, and regulate the Chong and Ren meridians.

[0103] Prescription: Nourishing Essence and Soothing Liver Decoction. One dose of the above Chinese herbs daily. Decoct to obtain 300mL of liquid, divided into two warm doses, 150mL each time, morning and evening. One course of treatment is 28 days, with three consecutive courses. After the first course (November 17, 2023), menstruation did not occur; however, symptoms such as hot flashes, sweating, and irritability significantly decreased, vaginal discharge increased, sleep improved, and bowel movements normalized. Hormone re-examination: FSH: 50.16 mIU / mL, LH: 35.77 mIU / mL, E2: 80.1 pg / mL. After the second course (December 17, 2024), menstruation still did not occur, but occasional hot flashes and sweating occurred, irritability disappeared, vaginal discharge increased, sleep improved, and bowel movements normalized. Follow-up hormone tests: FSH: 35.16 mIU / mL, LH: 20.77 mIU / mL, E2: 105.1 pg / mL. After the third course of treatment (January 17, 2025), menstruation resumed on January 15, 2024, with very light flow and pale red color. Vaginal moisture and normal discharge. Appetite and sleep were good, and bowel movements were regular. Hormone tests: FSH: 16.23 mIU / mL, LH: 10.66 mIU / mL, E2: 88.7 pg / mL. Follow-up transvaginal ultrasound: Uterus anteverted, size: 4.0cm × 4.0cm × 3.2cm, En 0.6cm; Right ovary size: 2.3cm × 2.2cm, containing 3 follicles, the largest measuring 0.4cm × 0.5cm; Left ovary size: 2.6cm × 1.6cm, containing 1 very small follicle measuring 0.4cm × 0.2cm. During a six-month follow-up, menstruation became regular with moderate flow, and clinical symptoms largely disappeared. Ultrasound monitoring showed a significant increase in the number of antral follicles compared to before treatment, indicating that ovarian function had recovered and remained stable.

[0104] Case 4: Chen, female, 29 years old. Initial visit: May 15, 2024.

[0105] Chief complaint: Menstrual cycle delayed by 14 years.

[0106] Present Illness: The patient had her first menstruation at age 15, with irregular periods lasting 4-5 days / 28 days to every six months. Menstrual flow was moderate, bright red, with no clots or dysmenorrhea. At age 16, she was diagnosed with premature ovarian failure at another hospital and has been taking oral hormones (Femoston 2 / 10) to maintain her menstrual cycle. Last Menstrual Period: May 13, 2024 (after taking Femoston), M3, scanty menstrual flow, bright red, no clots or dysmenorrhea. She typically has a poor appetite, is easily awakened, has many dreams, and has loose stools twice daily. Physical Examination: Dark red tongue with a thin white coating and teeth marks on the edges; deep, thready, weak, and slightly hesitant pulse.

[0107] Obstetric history: Married for six months, G0, no pregnancy despite not using contraception, planning to conceive.

[0108] Physical and chemical examination (May 15, 2024): Gynecological examination: No abnormalities found. Sex hormone tests: FSH: 89.22 mIU / mL, LH: 48.53 mIU / mL, E2: 31.03 pmol / L. Vaginal ultrasound: Uterus anteverted, size: 3.1cm × 3.0cm × 2.9cm, endometrial thickness: 0.3cm; right ovary, size: 1.5cm × 1.2cm, no follicles seen, solid echo; left ovary, size: 1.8cm × 0.7cm, no obvious follicles seen.

[0109] Western medical diagnosis: premature ovarian failure.

[0110] Traditional Chinese Medicine diagnosis: Delayed menstruation (Kidney deficiency, liver stagnation, and spleen deficiency syndrome).

[0111] Treatment: Tonify the kidneys and replenish essence, soothe the liver and strengthen the spleen, and regulate the Chong and Ren meridians.

[0112] Prescription: Instruct the patient to immediately discontinue oral Femoston. Administer Yangjing Shugan Decoction. One dose of the above Chinese medicine is prescribed daily. Decoct to obtain 300mL of liquid, divided into two warm doses of 150mL each, morning and evening. One course of treatment is 28 days, with three consecutive courses. After the first course (June 14, 2024), menstruation did not occur, but vaginal discharge increased, sleep improved, appetite improved, and bowel movements were loose twice daily. Follow-up sex hormone tests showed: FSH: 57.24 mIU / mL, LH: 33.53 mIU / mL, E2: 78.5 pmol / L, indicating improved ovarian function. After the second course (July 15, 2024), menstruation still did not occur, discharge continued to increase, appetite and sleep were good, and bowel movements decreased to once daily. Follow-up sex hormone tests: FSH: 47.24 mIU / mL, LH: 23.53 mIU / mL, E2: 68.3 pmol / L. Follow-up vaginal ultrasound: Uterus size same as before, En 0.5cm; right ovary size: 1.8cm × 1.7cm, with one follicle visible inside, approximately 0.2cm × 0.1cm in size; left ovary size: 1.8cm × 1.5cm, no follicle seen inside. After the third course of treatment (August 15, 2024), menstruation occurred naturally on August 13, 2024, with very light flow, brownish color, and stopped on the same day. Appetite was good, sleep was fair, and bowel movements were normal. Follow-up sex hormone tests: FSH: 22.72 mIU / mL, LH: 18.35 mIU / mL, E2: 108.3 pmol / L. Follow-up transvaginal ultrasound: Uterus size 3.5cm × 3.6cm × 2.9cm, En 0.5cm; Right ovary size: 2.2cm × 1.2cm, containing one follicle measuring 0.7cm × 0.5cm and 0.6cm × 0.5cm; Left ovary size: 1.8cm × 0.7cm, containing what appears to be one follicle measuring 0.3cm × 0.2cm. These findings suggest significant recovery of ovarian function. During a six-month follow-up, the patient established an independent menstrual cycle (3-4 / 40-65 days), with normal menstrual flow, requiring no exogenous hormone support, and clinical indicators remained stable.

[0113] Example 3

[0114] Component analysis will be conducted on the compound drug for treating premature ovarian failure.

[0115] The compound formula of Yangjing Shugan Decoction was analyzed using an ultra-high performance liquid chromatography-high resolution mass spectrometry (UPLC-Q-TOF / MS) system. The chromatographic column was an Agilent ZORBA×SB-Aq (2.1×100 mm, 1.8 µm). The flow rate was set to 0.3 mL / min, and the injection volume was 2 μL. The mobile phase consisted of acetonitrile (A) and 0.1% formic acid aqueous solution (B). The multi-step linear elution gradient program was as follows: 0–5 min, 0–10% A; 5–13 min, 10–15% A; 13–24 min, 15–28% A; 24–30 min, 28–70% A; 30–33 min, 70–95% A; 33–36 min, 95% A; 36–36.1 min, 95–0% A; 36.1–39 min, 0% A.

[0116] An AB Scie×Triple TOF® 4600 high-resolution mass spectrometry system (SCIE) was used with AnalystTF 1.7.1 and PeakView 1.2 software. MS and MS / MS data were acquired based on the electrospray ionization source in negative / positive ion mode (ESI-Negative / Positive). The mass spectrometry parameters were set as follows: TOF mass scan range: 50–1700; ion source gas 1: 50 psi; ion source gas 2: 50 psi; curtain gas: 35 psi; ion spray voltage fluctuation: -4500 / 5000 V; ion source temperature: 500℃; declustering voltage: 100 V; collision energy: 10 eV. MS / MS mass scan range: 50–1250; declustering voltage: 100 V; collision energy: ±40 eV; collision energy broadening: 20 eV; ion release delay: 30 ms; ion release width: 15 ms.

[0117] To identify the main components of YJSGD, we used ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) to analyze the chemical composition of YJSGD. Dual detection in positive and negative ion modes ensured comprehensive analysis of the ZSD extract. Figure 1Based on a comprehensive analysis of high-resolution mass spectrometry databases of natural products and relevant literature, combined with full-scan mode, primary mass spectrometry ion data, secondary collision energy, and fragment characteristics, 10 compounds in YJSGD were finally identified (Table 3). These components include: rehmannia glycoside D (CAS: 81720-08-3), monoglobin (CAS: 25406-64-8), loganin (CAS: 18524-94-2), paeoniflorin (CAS: 23180-57-6), ferulic acid (CAS: 1135-24-6), hyperoside (CAS: 482-36-0), privetin (CAS: 449733-84-0), hesperidin (CAS: 520-26-3), luteolin (CAS: 491-70-3), and ursolic acid (CAS: 77-52-1).

[0118] Example 4

[0119] Pharmacological experiments will be conducted on a compound drug for treating premature ovarian failure.

[0120] 1. Establishment of animal models

[0121] 1.1 Laboratory Animals

[0122] Sixty 6-week-old female (SPF) grade C57BL / 6J mice, weighing 20±2g, were housed in an environment with a 12-hour light / dark cycle, a temperature of 22-25℃, and a humidity of 40%-70%. After 3 days of acclimatization, they were randomly divided into 6 groups: model group, blank group, positive group, and high, medium, and low dose treatment groups.

[0123] 1.2 Model Construction

[0124] Except for the control group, aging mouse models were established by continuous intraperitoneal injection of D-galactose (D-galactose, 0.2 mL, 1.35 g / kg) for 42 days. Mice in the control group and model group were administered physiological saline (15 mL / kg, 0.1 mL) by gavage daily; mice in the positive control group were administered vitamin C (100 mg / kg, 0.1 mL) by gavage daily; mice in the essence-nourishing and liver-soothing group were administered essence-nourishing and liver-soothing decoction by gavage daily, with low dose (25 mg / kg, 0.22 mL), medium dose (50 mg / kg, 0.22 mL), and high dose (100 mg / kg, 0.22 mL); mice in the essence-nourishing and liver-soothing group were administered essence-nourishing and liver-soothing decoction by gavage daily, with low dose (25 mg / kg, 0.22 mL), medium dose (50 mg / kg, 0.22 mL), and high dose (100 mg / kg, 0.22 mL).

[0125] 1.3 Sample collection: After the administration of the drug, blood was collected from the eyeballs, and the ovaries were removed and weighed.

[0126] 1.4 Index Measurement:

[0127] ① Calculate the ovarian index using the following formula [Ovarian index = ovarian weight (g) / animal body weight (kg)];

[0128] ② The serum levels of E2, FSH, and LH in each group of mice were measured;

[0129] ③ Determine the activities of SOD, CAT, GSH-Px and MDA content in the serum of mice in each group.

[0130] ④ Measure the serum levels of IL-6, IL-1β, and TNF-α in each group of mice.

[0131] In this embodiment, a POF mouse model was constructed by continuous injection of D-galactose. Figure 1 (B) and monitored changes in mouse body weight throughout the experiment. Figure 2 As shown, during the entire experiment, the body weight of the control group mice steadily increased, while the body weight gain of the model group mice was slower than that of the control group. After treatment with Yangjing Shugan Decoction, the body weight gain trend of the model mice gradually became consistent with that of the control group. As shown in Table 3, compared with the control group, the organ index of the model group mice was significantly reduced (p<0.05); after intervention with Yangjing Shugan Decoction, the organ index of the mice rebounded, with the 50 mg / kg (YJSGD-M) dose showing the most significant improvement (Table 1).

[0132] Table 1. Effects of Yangjing Shugan Decoction on organ index in mice with ovarian aging (Mean±SD, n=10)

[0133]

[0134] like Figure 3 As shown, compared with the control group, the FSH and LH levels in the model group mice were significantly increased (p<0.001), while the E2 level and LH concentration were significantly decreased (p<0.001). In contrast, after YJSGD administration, the serum FSH level in POF mice was significantly decreased (p<0.01, p<0.05), while the E2 and LH levels were significantly increased (p<0.01, p<0.05). Overall, YJSGD effectively improved the estrous cycle disorder in POF mice and reversed the abnormal hormone levels, with the YJSGD-M group showing the best effect, indicating that YJSGD can effectively improve ovarian function in POF mice.

[0135] like Figure 4As shown in A-4D, compared with the control group, the serum MDA level in the model group mice was significantly increased (p<0.001), while the activities of CAT, SOD, and GSH-Px were significantly decreased (p<0.05, p<0.001). Intervention with Yangjing Shugan Decoction significantly reduced MDA levels in POF mice and increased the activities of CAT, SOD, and GSH-Px (p<0.05, p<0.01). Notably, the antioxidant activity of the medium-dose group of Yangjing Shugan Decoction was significantly enhanced (p<0.05, p<0.01). These results indicate that Yangjing Shugan Decoction (especially the medium dose) effectively improved D-galactose-induced oxidative damage in POF mice by upregulating antioxidant enzyme activity and inhibiting lipid peroxidation.

[0136] like Figure 5 As shown in A-5C, compared with the control group, the serum levels of IL-1β, IL-6, and TNF-α in the model group mice were significantly increased (p<0.01). After intervention with medium and high doses of Yangjing Shugan Decoction, the levels of IL-1β and IL-6 decreased significantly (p<0.05, p<0.01). Notably, the TNF-α level in the medium-dose YJSGD intervention group was also significantly inhibited (p<0.05).

[0137] 1.5 Gut Microbiota Analysis

[0138] Approximately 0.5 grams of fecal samples were collected from different groups for subsequent analysis. To detect differences in gut microbiota (GM) among samples, we performed alpha and beta diversity analyses. For alpha diversity, “Vegan” was used to assess the richness and diversity of each sample, and beta diversity was assessed using principal coordinate analysis (PCoA) and similarity analysis (ANOSIM) based on the Bray-Curtis difference index.

[0139] 1.5.1 YJSGD altered the gut microbiota composition of POF mice

[0140] Gut microbiota diversity was analyzed using single-molecule real-time sequencing technology. Results are as follows: Figure 6 C-6D analysis showed that the ACE and Chao1 indices of the YJSGD-M group were significantly higher than those of the model group (P<0.01), indicating higher species richness in this group. Furthermore, beta diversity analysis based on PCA and PCoA revealed a significant separation between the microbial community structure of the YJSGD-M group and the model group (6G-6H). Specifically, PC1 and PC2 in the PCA group explained 21.97% and 16.93% of the variation, respectively, while PCoA explained 9.79% and 7.86%, respectively. These results confirm that YJSGD-M can significantly alter the gut microbiota composition of POF mice.

[0141] like Figure 7As shown, at the phylum level, Firmicutes and Bacteroidetes were the dominant bacterial groups in the control group. Compared with the control group, the F / B ratio in the model group mice was significantly increased, the relative abundance of Desulfobacterota increased, while the relative abundance of Proteobacteria and Verrucomicrobia decreased. After YJSGD intervention, the F / B ratio in POF mice was significantly reduced, the abundance of Desulfobacterota decreased, and the abundance of Actinobacteria increased. These results suggest that YJSGD can effectively reverse the phylum-level imbalance of gut microbiota induced by POF.

[0142] like Figure 8 As shown, at the genus level, compared with the control group, the relative abundance of beneficial bacteria *Ligilactobacillus*, *unclassified_Muribaculaceae*, *Lactobacillus*, *Bacteroides*, *Parabacteroides*, and *Muribaculum* decreased in the model group mice, while the relative abundance of harmful bacteria *Parasutterella*, *Alloprevotella*, and *unclassified_Desulfovibrionaceae* increased. Compared with the model group mice, the relative abundance of beneficial bacteria *Lactobacillus*, *Akkermansia*, *Dubosiella*, and *Ileibacterium* increased in the YJSGD group, while the relative abundance of harmful bacteria such as *Alloprevotella* and *unclassified_Desulfovibrionaceae* decreased. These results indicate that YJSGD can improve the diversity and richness of the gut microbiota.

[0143] At the genus level, the model group showed a significant reduction in beneficial bacteria compared to the control group, including *Ligilactobacillus*, *unclassified_Muribaculaceae*, *Lactobacillus*, *Bacteroides*, *Parabacteroides*, and *Muribaculum*; while harmful bacteria were significantly enriched, such as *Parasutterella*, *Alloprevotella*, and *unclassified_Desulfovibrionaceae*. After YJSGD intervention, the abundance of beneficial bacteria such as *Lactobacillus*, *Akkermansia*, *Dubosiella*, and *Ileibacterium* rebounded, while the abundance of the aforementioned harmful bacteria decreased. This indicates that YJSGD restores the balance of the gut microbiota by precisely regulating key bacterial genera.

[0144] 1.6 Determination of short-chain fatty acid content in mouse feces

[0145] The levels of short-chain fatty acids (SCFAs) in mouse feces were determined by gas chromatography-mass spectrometry (GC-MS). A Trace1310-TSQ800 GC-MS system equipped with a TR-FFAP column (30 mm × 0.25 mm × 0.25 μm) was used to analyze the SCFA content. A flame ionization detector (FID) was used, with an injection port temperature of 230 °C, an injection volume of 1.0 μL, and an ion source temperature of 280 °C. The temperature program was as follows: initial temperature 100 °C, held for 1 min; increased to 160 °C at a rate of 5 °C / min, held for 5 min; then increased to 200 °C at a rate of 25 °C / min, held for 2 min. Helium was used as the carrier gas at a flow rate of 1.0 mL / min, with a split ratio of 10:1.

[0146] 1.6.1 YJSGD increased the level of fecal SCFAs in POF mice.

[0147] SCFAs are metabolites produced by the fermentation of dietary fiber such as polysaccharides by the gut microbiota. They can affect intestinal motility, enhance intestinal barrier function, and host metabolism. The total ion chromatogram of mixed-standard fatty acids and the content of short-chain fatty acids in the feces of mice in each group are shown in the table below. The results showed that acetic acid, propionic acid, and butyric acid were the main components of SCFAs in the feces of mice in each group, while the contents of isobutyric acid, valeric acid, and isovaleric acid were low. Compared with the control group, the SCFA content of mice in the model group was significantly reduced (P<0.05). After YJSGD intervention, the levels of acetic acid, propionic acid, butyric acid, isobutyric acid, valeric acid, and isovaleric acid all increased (P<0.05).

[0148] Table 2. Analysis of fecal SCFAs levels

[0149] 1.7 Non-targeted metabolomics analysis of mouse serum

[0150] Targeted metabolomics analysis of mouse serum was performed according to the method of Li et al. 100 µL of serum was taken, 300 µL of methanol was added, vortexed for 30 s, centrifuged at 12000 rpm / min for 15 min, the supernatant was dried under nitrogen, reconstituted with 1 mL of ultrapure water, vortexed for 15 s, and centrifuged again (12000 rpm / min, 15 min). The sample was then filtered through a 0.22 µm filter before injection. QC samples were prepared by mixing equal volumes of each serum sample.

[0151] Chromatographic analysis was performed using a Thermo Golden C18 column (50 × 2.1 mm, 1.9 µm) at 35 °C and a sample tray temperature of 4 °C. Mobile phase: A was 0.1% formic acid in water, and B was 0.1% formic acid in acetonitrile; flow rate: 0.3 mL / min. Gradient program: 0–1 min, 3% B; 1–16 min, 3%–68% B; 16–18 min, 68% B; 18–25 min, 68%–90% B; 25–26 min, 90%–100% B; 26–32 min, 100% B. Injection volume: 2.0 µL.

[0152] Mass spectrometry was performed using an electrospray ionization (ESI) source in both positive and negative ion modes, with full MS / dd-MS2 (TopN=5) scans. The spray voltage was 3.5 kV, the capillary temperature was 320 °C, and the auxiliary gas and heater temperatures were 350 °C. Collision energies were 10%, 20%, and 40%. Data were processed using ×calibur software. Samples were randomly analyzed by UHPLC-ESI-QE×active Orbitrap / MS, interspersed with blanks (50% acetonitrile-water) and QC samples to monitor system stability.

[0153] 1.7.1 Analysis of differences in metabolites between groups

[0154] First, the raw mass spectrometry data of each group were preprocessed, including noise reduction, baseline correction, normalization, and peak alignment, and then analyzed uniformly using SIMCA14.1 software. The PCA analysis results are shown in the figure. Most of the blood samples from the blank control group mice were close together, but never overlapped. Although there was very little overlap between groups, the distances between groups of different colors were significant, indicating clear differences in both negative and positive ion modes.

[0155] OPLS-DA analysis of differentially expressed metabolites in mouse blood showed significant separation between the blank control group and the model group, as well as between the model group and the medium-dose group, according to the scoring plot. This indicates that there are significant differences in metabolites between the groups. Figure 9 A-9F).

[0156] 1.7.2 Screening for Differential Metabolites

[0157] Non-targeted metabolomics technology, combined with bioinformatics methods, was used to screen differentially expressed metabolites and perform pathway enrichment analysis to accurately capture metabolic changes in POF mice. Based on the screening criteria of VIP>1 and P<0.05, differentially expressed metabolites were selected using ESI. + / ESI – A total of 18 common potential biomarkers were identified in the dual-ion mode (Table 3). The results showed that the compound significantly restored the levels of 18 differentially expressed metabolites that had changed in the model group.

[0158] Table 3 Information on common potential biomarkers

[0159]

[0160] Note: Blank control group (C), model group (M), medium-dose compound extract treatment group (M2). Trend: "↓" indicates downward adjustment, "↑" indicates upward adjustment.

[0161] 1.7.3 Metabolic pathway analysis

[0162] To investigate the effects of YJSGD on blood biomarkers and related metabolic pathways in POF model mice, MetaboAnalyst 5.0 was used to perform pathway enrichment analysis on differentially expressed metabolites. Figure 4 A). KEGG metabolic pathways were screened based on the criteria of VIP>1 and p<0.05, and pathway importance was assessed through topological analysis: the pathway impact was plotted on the x-axis, and enrichment significance (-log) was used to evaluate the pathway importance. 10 Plotting P as the ordinate ( Figure 4 B). The red circles in the upper right quadrant of the graph (with area and color depth increasing with significance) indicate key pathways, whose -log 10 A higher P-value indicates a stronger metabolic association between groups. The results showed that differentially metabolites were significantly enriched in two core pathways: steroid hormone biosynthesis and acrylonic acid metabolism (P<0.01), as well as beta-alanine metabolism and histidine metabolism.

Claims

1. A composition for nourishing essence and soothing the liver, used to treat premature ovarian failure of the kidney-deficiency, liver-stagnation, and spleen-deficiency type, characterized in that, The aforementioned Nourishing Essence and Soothing Liver Decoction is composed of 20-30 parts of Cornus officinalis, 10-20 parts of Rehmannia glutinosa, 5-15 parts of Angelica sinensis, 10-20 parts of Paeonia lactiflora, 10-20 parts of Dioscorea opposita, 5-15 parts of Citrus reticulata, 10-20 parts of Cuscuta chinensis, 10-20 parts of Ligustrum lucidum, 5-15 parts of Lycopus lucidus, and 10-20 parts of Cyperus rotundus, by weight.

2. The composition for nourishing essence and soothing the liver for treating premature ovarian failure of the kidney deficiency, liver stagnation, and spleen deficiency type according to claim 1, characterized in that, The aforementioned Nourishing Essence and Soothing Liver Decoction is composed of 25-30 parts of Cornus officinalis, 15-20 parts of Rehmannia glutinosa, 10-15 parts of Angelica sinensis, 15-20 parts of Paeonia lactiflora, 15-20 parts of Dioscorea opposita, 10-15 parts of Citrus reticulata, 15-20 parts of Cuscuta chinensis, 15-20 parts of Ligustrum lucidum, 10-15 parts of Lycopus lucidus, and 15-20 parts of Cyperus rotundus, by weight.

3. A composition for nourishing essence and soothing the liver for treating premature ovarian failure of the kidney-deficiency, liver-stagnation, and spleen-deficiency type, as described in claim 1 or 2, characterized in that... The aforementioned Nourishing Essence and Soothing Liver Decoction is composed of 25 parts Cornus officinalis, 16 parts Rehmannia glutinosa, 10 parts Angelica sinensis, 15 parts Paeonia lactiflora, 15 parts Dioscorea opposita, 10 parts Citrus reticulata, 15 parts Cuscuta chinensis, 15 parts Ligustrum lucidum, 10 parts Lycopus lucidus, and 15 parts Cyperus rotundus, according to the weight proportions.

4. A composition for nourishing essence and soothing the liver for treating premature ovarian failure of the kidney-deficiency, liver-stagnation, and spleen-deficiency type, as described in claim 1 or 2, characterized in that, The aforementioned Nourishing Essence and Soothing Liver Decoction is composed of 30 parts Cornus officinalis, 20 parts Rehmannia glutinosa, 15 parts Angelica sinensis, 15 parts Paeonia lactiflora, 15 parts Dioscorea opposita, 10 parts Citrus reticulata peel, 15 parts Cuscuta chinensis, 15 parts Ligustrum lucidum, 10 parts Lycopus lucidus, and 15 parts Cyperus rotundus, according to the weight proportions.

5. A method for preparing the essence-nourishing and liver-soothing composition according to claim 1 for treating premature ovarian failure of the kidney-deficiency, liver-stagnation, and spleen-deficiency type, characterized in that... The preparation method is as follows: Weigh out 20-30 parts of Cornus officinalis, 10-20 parts of Rehmannia glutinosa, 5-15 parts of Angelica sinensis, 10-20 parts of Paeonia lactiflora, 10-20 parts of Dioscorea opposita, 5-15 parts of Citrus reticulata, 10-20 parts of Cuscuta chinensis, 10-20 parts of Ligustrum lucidum, 5-15 parts of Lycopus lucidus, and 10-20 parts of Cyperus rotundus according to the following proportions: mix them together, add 8-10 times the amount of water, soak for half an hour, decoct for half an hour, filter out the liquid, add 10 times the amount of water, decoct for half an hour, filter out the liquid, repeat twice; combine the three liquids, concentrate, dry, and pulverize to obtain a dry extract powder.

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