1.Analysis of Active Components and Metabolites of Modified Ermiao Decoction Based on UPLC-Q-Exactive Orbitrap-MS/MS Technology
Xian GE ; Hongting ZHAO ; Li CHEN ; Ruoxi ZHANG ; Shichun ZHU ; Qingling REN
Journal of Nanjing University of Traditional Chinese Medicine 2025;41(8):998-1010
OBJECTIVE To conduct qualitative analysis of chemical constituents in the crude extract of Modified Ermiao Formu-la,along with its blood-absorbed components and metabolites in rats post-administration employing ultra-performance liquid chroma-tography coupled with quadrupole-exactive orbitrap tandem mass spectrometry(UPLC-Q-Exactive Orbitrap-MS/MS).METHODS Separation was performed on a Waters HSS T3 column(100 mm×2.1 mm,1.8 μm)using gradient elution with 0.1%formic acid in water(mobile phase A)and 0.1%formic acid in acetonitrile(mobile phase B),under controlled conditions:column temperature maintained at 40℃,flow rate set to 0.3 mL·min-1,and injection volume fixed at 2 μL.Mass spectrometric data acquisition utilized an electrospray ionization(ESI)source with scanning in both positive and negative ion modes.RESULTS By analyzing precise mo-lecular weights,retention times,and MS/MS fragmentation patterns,and cross-referencing these against established databases and published literature,173 chemical constituents were definitively identified in the Modified Ermiao Formula crude extract.These prima-rily comprised flavonoids,organic acids,and alkaloids.Additionally,32 prototype components and 13 metabolites were characterized in the serum of dosed rats.Organic acids constituted the predominant class of serum prototype components,undergoing in vivo metabo-lism principally through demethylation and carboxyl glucuronidation.CONCLUSION This study provides the initial delineation of blood-absorbed prototype components derived from Modified Ermiao Formula,with concomitant identification of their metabolites,thereby establishing a foundational framework for elucidating the pharmacologically active constituents of this formula.
2.Exploration on the Method of Soothing Liver and Strengthening Spleen in Treating Polycystic Ovary Syndrome from the Theory of"Simultaneous Upper and Lower Diseases,Treating the Middle Energizer"
Chinese Journal of Information on Traditional Chinese Medicine 2025;32(11):25-29
Polycystic ovary syndrome(PCOS)is one of the main causes of ovulatory dysfunction-induced female infertility.The dysfunction of the hypothalamic-pituitary-ovarian axis(HPOA)is a key factor promoting reproductive damage and metabolic abnormalities in PCOS.This article explored the correlation between HPOA dysfunction and the pathogenesis of heart-kidney disharmony in TCM based on Ye Tianshi's theory of"simultaneous upper and lower diseases,treating the middle energizer".It is believed that the treatment of PCOS should aim at restoring the ascending and descending of qi in the middle energizer,with the main therapeutic approach being soothing the liver and strengthening the spleen,which can provide ideas for the clinical treatment of PCOS.
3.Mechanisms of Wandai Decoction in Improving Vaginal Flora of Vulvovaginal Candidiasis of the Spleen Deficiency and Excessive Dampness Type:A Study Based on Metagenomics and Metabolomics
Yue HAN ; Su SHEN ; Jie FU ; Qingling REN
Journal of Sichuan University (Medical Sciences) 2025;56(1):68-73
Objective To explore the mechanism by which Wandai Decoction prevents and treats vulvovaginal candidiasis(VVC)of the spleen deficiency and excessive dampness type and restores the vaginal flora structure,and to identify the potential metabolic pathways involved using metagenomics and metabolomics.Methods Twenty VVC patients who met the inclusion criteria were randomly assigned to a Wandai Decoction group and a fluconazole group(n=10 in each group).Subjects in the fluconazole group were given a single oral dose of 150 mg fluconazole,while those in the Wandai Decoction group took the Wandai Decoction orally for 14 days.The vulvovaginal signs and symptoms(VSS)scores of both patient groups were evaluated before and after treatment.Vaginal secretions were collected before and after treatment.The Illumina sequencing and the liquid chromatography with tandem mass spectrometry(LC-MS/MS)platform were used to conduct metagenomic and metabolomics analyses of the vaginal secretions,respectively.Results The VSS score results showed that the VSS scores of both groups decreased after treatment compared with those before treatment(P<0.01),and there was no statistically significant difference in the VSS scores between the two groups after treatment.Metagenomics results showed that,after treatment,the vaginal microbial communities in the Wandai Decoction group were of CST Ⅱ and Ⅴ types(predominated by Lactobacillus gasseri and Lactobacillus jensenii),while those in the fluconazole group were Lactobacillus_intestinalis and Streptococcus_sp._oral_taxon_431.KEGG functional enrichment analysis results showed that,in terms of the cell cycle and meiosis functions of Candida albicans,statistically significant differences between the Wandai Decoction and fluconazole groups were observed(P<0.05).Metabolomic analysis identified 120 differential metabolites between the two groups after treatment.The results of KEGG metabolic pathway enrichment analysis of differential metabolites showed that the Wandai Decoction might be significantly superior to fluconazole in improving local vaginal metabolic pathways of α-linolenic acid,glycerophospholipid metabolism,pentose and glucuronic acid interconversion,and arachidonic acid.Conclusion The Wandai Decoction can improve the vaginal flora of VVC patients.It may be superior to fluconazole in the signaling pathways of the cell cycle and meiosis.The improvement of the vaginal flora by the Wandai Decoction may be associated with its effect on metabolic pathways of glycerophospholipid metabolism,pentose and glucuronic acid interconversion,and others in the vagina.
4.Analysis of Active Components and Metabolites of Modified Ermiao Decoction Based on UPLC-Q-Exactive Orbitrap-MS/MS Technology
Xian GE ; Hongting ZHAO ; Li CHEN ; Ruoxi ZHANG ; Shichun ZHU ; Qingling REN
Journal of Nanjing University of Traditional Chinese Medicine 2025;41(8):998-1010
OBJECTIVE To conduct qualitative analysis of chemical constituents in the crude extract of Modified Ermiao Formu-la,along with its blood-absorbed components and metabolites in rats post-administration employing ultra-performance liquid chroma-tography coupled with quadrupole-exactive orbitrap tandem mass spectrometry(UPLC-Q-Exactive Orbitrap-MS/MS).METHODS Separation was performed on a Waters HSS T3 column(100 mm×2.1 mm,1.8 μm)using gradient elution with 0.1%formic acid in water(mobile phase A)and 0.1%formic acid in acetonitrile(mobile phase B),under controlled conditions:column temperature maintained at 40℃,flow rate set to 0.3 mL·min-1,and injection volume fixed at 2 μL.Mass spectrometric data acquisition utilized an electrospray ionization(ESI)source with scanning in both positive and negative ion modes.RESULTS By analyzing precise mo-lecular weights,retention times,and MS/MS fragmentation patterns,and cross-referencing these against established databases and published literature,173 chemical constituents were definitively identified in the Modified Ermiao Formula crude extract.These prima-rily comprised flavonoids,organic acids,and alkaloids.Additionally,32 prototype components and 13 metabolites were characterized in the serum of dosed rats.Organic acids constituted the predominant class of serum prototype components,undergoing in vivo metabo-lism principally through demethylation and carboxyl glucuronidation.CONCLUSION This study provides the initial delineation of blood-absorbed prototype components derived from Modified Ermiao Formula,with concomitant identification of their metabolites,thereby establishing a foundational framework for elucidating the pharmacologically active constituents of this formula.
5.Exploring the Role of Glucose Metabolism Reprogramming in Regulating Polycystic Ovary Syndrome Based on the Theory of"Spleen Qi to Dispersing Essence"
Tingting SHANG ; Wenyue CHEN ; Die WU ; Chenxi GU ; Shu ZOU ; Qingling REN
World Science and Technology-Modernization of Traditional Chinese Medicine 2025;27(3):878-886
Polycystic ovary syndrome(PCOS)is a complex disease that significantly impacts women's reproductive and metabolic health.The reprogramming of glucose metabolism points to fundamental changes in energy metabolism,which are central to the imbalance in energy metabolism,hormonal abnormalities,increasing insulin resistance,and chronic inflammatory states observed in PCOS.This represents a core pathological link in the pathogenesis of PCOS.The theory of"Spleen Qi to Dispersing Essence"provides a comprehensive overview of spleen function,essential for ensuring the normal operation of organs and the distribution of Qi,blood,and bodily fluids.It serves as a crucial theoretical support for the treatment of PCOS with traditional Chinese medicine.This article explores the role of regulating PCOS based on the theory of"Spleen Qi to Dispersing Essence"with a focus on the reprogramming of glucose metabolism.The mechanism of traditional Chinese medicinal treatments that assist Spleen Qi to Dispersing Essence is closely related to regulating glucose metabolism reprogramming,offering a novel research direction for the treatment of PCOS with TCM.
6.Exploration on the Method of Soothing Liver and Strengthening Spleen in Treating Polycystic Ovary Syndrome from the Theory of"Simultaneous Upper and Lower Diseases,Treating the Middle Energizer"
Chinese Journal of Information on Traditional Chinese Medicine 2025;32(11):25-29
Polycystic ovary syndrome(PCOS)is one of the main causes of ovulatory dysfunction-induced female infertility.The dysfunction of the hypothalamic-pituitary-ovarian axis(HPOA)is a key factor promoting reproductive damage and metabolic abnormalities in PCOS.This article explored the correlation between HPOA dysfunction and the pathogenesis of heart-kidney disharmony in TCM based on Ye Tianshi's theory of"simultaneous upper and lower diseases,treating the middle energizer".It is believed that the treatment of PCOS should aim at restoring the ascending and descending of qi in the middle energizer,with the main therapeutic approach being soothing the liver and strengthening the spleen,which can provide ideas for the clinical treatment of PCOS.
7.Exploring the Role of Glucose Metabolism Reprogramming in Regulating Polycystic Ovary Syndrome Based on the Theory of"Spleen Qi to Dispersing Essence"
Tingting SHANG ; Wenyue CHEN ; Die WU ; Chenxi GU ; Shu ZOU ; Qingling REN
World Science and Technology-Modernization of Traditional Chinese Medicine 2025;27(3):878-886
Polycystic ovary syndrome(PCOS)is a complex disease that significantly impacts women's reproductive and metabolic health.The reprogramming of glucose metabolism points to fundamental changes in energy metabolism,which are central to the imbalance in energy metabolism,hormonal abnormalities,increasing insulin resistance,and chronic inflammatory states observed in PCOS.This represents a core pathological link in the pathogenesis of PCOS.The theory of"Spleen Qi to Dispersing Essence"provides a comprehensive overview of spleen function,essential for ensuring the normal operation of organs and the distribution of Qi,blood,and bodily fluids.It serves as a crucial theoretical support for the treatment of PCOS with traditional Chinese medicine.This article explores the role of regulating PCOS based on the theory of"Spleen Qi to Dispersing Essence"with a focus on the reprogramming of glucose metabolism.The mechanism of traditional Chinese medicinal treatments that assist Spleen Qi to Dispersing Essence is closely related to regulating glucose metabolism reprogramming,offering a novel research direction for the treatment of PCOS with TCM.
8.Clinical Efficacy of Wandai Decoction in Preventing and Treating VVC with Spleen Deficiency-Dampness Abundance Syn-drome and Its Impact on DNA Damage
Yue HAN ; Su SHEN ; Jie FU ; Qingling REN
Journal of Nanjing University of Traditional Chinese Medicine 2024;40(2):190-195
OBJECTIVE To explore the clinical efficacy of Wandai Decoction in the treatment of vulvovaginal candidiasis(VVC)with spleen deficiency-dampness abundance and its effect on DNA damage.METHODS 70 VVC patients with spleen deficiency-dampness abundance who met the inclusion criteria were randomly divided into Wandai Decoction and fluconazole groups,35 cases each.The fluconazole group took 150 mg fluconazole orally once;the Wandai Decoction group took Wandai Decoction orally for 14 d.After treatment,changes in TCM syndrome scores of the two groups of patients were compared;the clinical cure(test of cure,TOC)rate and clinical improvement(CI)rate of the two groups of patients were evaluated;8-hydroxydeoxyguanosine(8-OHDG)in the vaginal lavage fluid was detected by colorimetry to assess DNA damage;clinical complete remission(follow up,FU),mycological negative conversion and recurrence rate were evaluated 3 months after treatment.RESULTS After treatment,the TCM syndrome scores of the two groups of patients were improved to varying degrees(P<0.05,P<0.01);the Wandai Decoction group was superior to the fluconazole group(P<0.05,P<0.01);there was no significant difference in TOC,CI,mycology negative conversion between the Wandai Decoction group and the fluconazole group(P>0.05);however,the FU and recurrence rate of the Wandai Decoction group were significantly better than those of the fluconazole group(P<0.05,P<0.01).After treatment,the expression of 8-OHDG in the vaginal lavage fluid of the fluconazole group increased significantly(P<0.001),while there was no significant change in the Wandai Decoction group,which was significantly lower than that of the fluconazole group(P<0.001).CONCLUSION The overall clinical efficacy of Wandai Decoction is equivalent to that of fluconazole,but it is better than fluconazole in improving TCM syndromes and pre-venting recurrence,with the advantage of not aggravating DNA damage in vaginal cells.
9.Role of Macrophage Polarization in Chronic Low-grade Inflammation of PCOS Based on Theory of Spleen Being in Charge of Defensive Function
Tingting SHANG ; Zixun ZHUANG ; Hua MA ; Bingying FAN ; Qingling REN
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(9):211-218
Polycystic ovary syndrome (PCOS) is a reproductive endocrine disorder characterized by coexisting reproductive dysfunction and glucolipid metabolic disturbance, affecting 8%-13% of women of reproductive age and 3%-11% of adolescent females. Due to the highly heterogeneous clinical features, symptom-oriented individualized strategies are commonly adopted for the treatment of PCOS. Chronic low-grade inflammation is one of the core mechanisms for the occurrence of PCOS. Macrophages, as foundational cells of innate immunity, play an indispensable role in modulating systemic inflammatory responses. The imbalance of macrophage M1/M2 polarization is involved in chronic low-grade inflammation in PCOS via pathways such as activating pro-inflammatory responses, disrupting ovarian tissue repair, stimulating excessive synthesis of androgens, and promoting the occurrence of insulin resistance. Reshaping the phenotype of macrophages might serve as a potential therapeutic strategy for PCOS. Traditional Chinese medicine (TCM) holds that spleen deficiency and phlegm dampness is a crucial pathogenesis of PCOS. The spleen, being in charge of defensive function, plays a key role in ensuring normal physiological functions such as transportation and defense against external pathogen during the occurrence and development of PCOS. The imbalance of macrophage polarization resembles the transition from spleen being in charge of defensive function to spleen losing its defensive role in TCM. Therefore, this paper, for the first time, explores the deep connection between macrophage polarization and the pathogenesis of chronic low-grade inflammation in PCOS from the TCM theory of spleen being in charge of defensive function, providing theoretical support and new research directions for the treatment and drug research of PCOS.
10.Effect and Mechanisms of Kaiyu Zhongyutang on Insulin Resistance, Glucose and Lipid Metabolism, Psychological State, and Embryo Outcome in Infertile Patients with PCOS Due to Liver Depression and Kidney Deficiency
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(24):130-137
ObjectiveTo explore the effects and mechanisms of Kaiyu Zhongyutang on insulin resistance, glucose and lipid metabolism, psychological state, and embryo outcome in the infertile patients with polycystic ovary syndrome (PCOS) due to liver depression and kidney deficiency. MethodThe 126 infertile patients with PCOS due to liver depression and kidney deficiency who underwent in vitro fertilization-embryo transfer (IVF-ET) in the Department of Reproduction of the affiliated hospital of Nanjing university of Chinese medicine were randomly assigned into the observation (Kaiyu Zhongyutang + metformin) and control (metformin) groups. The two groups were compared in terms of traditional Chinese medicine (TCM) syndrome scores, body mass index (BMI), glucose and lipid metabolism, homeostasis model assessment-insulin resistance (HOMA-IR), interleukin (IL)-6, IL-8, C-reactive protein (CRP), self-rating depression scale (SDS) score, self-rating anxiety scale (SAS) score, and clinical and laboratory scores of IVF after treatment. Result① After treatment, the observation group showed decreased scores of primary and secondary TCM syndromes and total TCM syndrome score (P < 0.05), and the control group presented decreased scores of irritability and depression and total TCM syndrome score (P<0.05). Compared with the control group, the observation group demonstrated reduced primary and secondary syndrome scores and total score after treatment (P<0.05). ② After treatment, both groups showed decreased BMI, lowered levels of fasting insulin (FINS), fasting plasma glucose (FPG), cholesterol (TC), triglyceride (TG), low-density lipoprotein (LDL), and HOMA-IR (P<0.05), and elevated high-density lipoprotein (HDL) level (P<0.05). Moreover, the observation group had lower BMI, FINS, FPG, TC, TG, LDL, and HOMA-IR and higher HDL than the control group (P<0.05). ③ The treatment in both groups lowered the levels of IL-6, IL-8, and CRP and decreased SDS and SAS scores (P<0.05). Moreover, the declines in the observation group were more obvious than those in the control group (P<0.05). ④ Correlation analysis before treatment, IL-6, IL-8, and CRP had positive correlations with BMI, HOMA-IR, TC, TG, LDL, SDS, and SAS (P<0.05) and a negative correlation with HDL (P<0.05). ⑤ The observation group showed reduced gonadotropin (Gn) using days and total Gn dose and higher two-pronuclear (2PN) fertilized oocytes, 2PN cleavage rate, normal fertilization rate, D3 transferable embryo number, D3 high-quality embryo number, high-quality embryo rate, and blastocyst formation rate than the control group (P<0.05). ConclusionKaiyu Zhongyutang can treat PCOS patients by improving the emotional and reproductive functions and alleviating insulin resistance and glucose and lipid metabolism disorders. Moreover, it can reduce the Gn dose and Gn using days in the IVF process, improve the quality and maturity of eggs, increase the egg fertilization rate, enhance the potential of embryo development, and increase the rate of blastocyst formation by inhibiting inflammation.

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