ObjectiveTo optimize and validate the optimal sequential alcohol-water extraction process of modified Banxia Xiexintang（MBXT） based on pharmacodynamic evaluation， combined with the G1-entropy weight method and Box-Behnken response surface methodology， and to systematically and comprehensively analyze the material basis of this formula， providing a scientific basis for its quality control and industrial production. MethodsRats were randomly divided into the normal group， model group， metformin group， and MBXT water extraction， water extraction and ethanol precipitation， sequential ethanol-water extraction groups. Except for the normal group， a polycystic ovary syndrome with insulin resistance（PCOS-IR） model was established in all rats via a high-fat diet combined with letrozole induction. Enzyme-linked immunosorbent assay（ELISA） biochemical assay kits and hematoxylin-eosin（HE） staining were used to compare sex hormone levels in serum and ovarian histopathology， thereby screening extraction process routes. Based on this， a comprehensive score was constructed using the G1-entropy weight method based on the transfer rates of index components（berberine hydrochloride and baicalin） and the dry extract rate. Box-Behnken response surface methodology was then utilized to optimize the extraction process parameters. Finally， the chemical constituents of the sample from the optimal process were qualitatively analyzed by ultra-performance liquid chromatography-quadrupole-electrostatic field orbitrap mass spectrometry（UPLC-Q-Orbitrap-MS）. ResultsPharmacodynamic findings revealed that compared with the normal group， serum testosterone（T） and luteinizing hormone（LH） levels were significantly elevated in the model group， while estradiol（E₂） and follicle-stimulating hormone（FSH） levels were significantly decreased（P<0.01）， with polycystic changes observed in ovarian tissues. Compared with the model group， all treatment groups significantly reversed the changes in sex hormone levels， with the sequential ethanol-water extraction group showing the optimal effect in improving the aforementioned indicators and pathological morphology， followed by subsequent process optimization. The optimized process involved adding 12 times the amount of 70% ethanol for extracting twice， each lasting 120 min， and adding 12 times the amount of water for extracting thrice， each lasting 90 min. Validation test results showed that under optimal process conditions， the average transfer rates of berberine hydrochloride and baicalin were 76.05% and 93.38%， respectively. MS analysis identified a total of 377 compounds， including 112 flavonoids， 41 terpenoids， 28 organic acids， 22 coumarins， and 8 alkaloids， while elucidating the cleavage patterns of key components. ConclusionThe optimized sequential ethanol-water extraction process is stable and feasible， effectively preserving the material basis of MBXT for treating PCOS-IR. It further clarifies the main chemical composition of this formula， providing a scientific basis for the development and quality control of its preparations.

ObjectiveTo evaluate the therapeutic effects of modified Banxia Xiexintang（MBXT） on polycystic ovary syndrome with insulin resistance（PCOS-IR） rats and reveal its potential mechanisms based on the integrated analysis of transcriptomics and metabolomics. MethodsFemale SD rats were selected， and a PCOS-IR model was established by intragastric administration of letrozole combined with a high-fat diet for 21 days. The modeled rats were randomly divided into the model group， MBXT low-， medium-， and high-dose groups（6.62， 13.23， 26.46 g·kg^-1）， and metformin group（0.158 g·kg^-1）， with a normal group set up separately. After 14 days of administration， the estrous cycle was observed， ovarian morphology was examined by hematoxylin-eosin（HE） staining， and the levels of testosterone（T）， estradiol（E₂）， follicle-stimulating hormone（FSH）， and luteinizing hormone（LH） in serum were detected by enzyme-linked immunosorbent assay（ELISA）. Serum metabolites and ovarian tissue gene expression were detected using ultra-performance liquid chromatography-quadrupole-electrostatic orbitrap mass spectrometry（UPLC-Q-Orbitrap-MS） and RNA-Seq technology， respectively， followed by multi-omics integrated analysis. ResultsPharmacodynamic findings revealed that all MBXT dose groups could reversed abnormal estrous cycles in PCOS-IR rats， improve polycystic ovarian lesions， and normalize dysregulated serum hormone levels（T， LH， E₂， FS， P<0.05， P<0.01）. Metabolomic analysis revealed that compared with the model group， MBXT reversed 278 differential metabolites such as estrone and S-formylglutathione， mainly involving pathways such as steroid hormone biosynthesis， glutathione metabolism， and lipid peroxidation regulation. Transcriptomic analysis identified 434 differentially expressed genes， and enrichment analysis revealed that MBXT significantly regulated lipid peroxidation defense systems， including glutathione metabolism， peroxisome function， and fatty acid metabolism， thereby intervening in ferroptosis processes. It also engaged in inflammation-related pathways such as the chemokine signaling pathway. Integrated analysis revealed that both metabolomics and transcriptomics co-enriched metabolic pathways associated with ferroptosis and fatty acid metabolism. And key Hub genes［such as Ras-related C3 botulinum toxin substrate 2 gene（Rac2） and Fas ligand gene（Faslg）］ showed significant correlations with differential metabolites. ConclusionMBXT can effectively ameliorate reproductive dysfunction and metabolic disorders in PCOS-IR rats. Its mechanism may be related to remodeling the immune-metabolism network， particularly by regulating MHC-mediated immune responses， inhibiting local ovarian ferroptosis， and enhancing steroid hormone synthesis pathways.

ObjectiveTo investigate the mechanism of modified Banxia Xiexintang（MBXT） in improving ovarian dysfunction in polycystic ovary syndrome with insulin resistance（PCOS-IR） rats by inhibiting ferroptosis through the adenosine monophosphate（AMP）-activated protein kinase（AMPK）/fatty acid synthase（FASN）/glutathione peroxidase 4（GPX4） signaling pathway. MethodsSeventy-six female SD rats were randomly divided into a normal group（n=13） and a modeling group（n=63）. The modeling group established a PCOS-IR model by intragastric administration of letrozole combined with a high-fat diet for 21 days. After successful modeling， these rats were randomly divided into the model group， MBXT low-， medium-， and high-dose groups（6.62， 13.23， 26.46 g·kg^-1）， metformin group（0.158 g·kg^-1）， and high-dose of MBXT combined with ferroptosis inducer Erastin group（15 mg·kg^-1）， with 10 rats in each group. After 14 days of intervention， ovarian pathological morphology was observed by hematoxylin-eosin（HE） staining， the mitochondrial ultrastructure of granulosa cells was observed by transmission electron microscopy（TEM）， ovarian reactive oxygen species（ROS） levels were detected by dihydroethidium（DHE） probe， biochemical methods were used to detect Fe²⁺， malondialdehyde（MDA）， glutathione（GSH） and other indicators in ovarian tissues， serum sex hormone and insulin levels were measured by enzyme-linked immunosorbent assay（ELISA）， and the protein expressions of AMPK， FASN， acyl-CoA synthetase long-chain family member 4（ACSL4）， GPX4， and solute carrier family 7 member 11（SLC7A11） in ovarian tissues were detected by Western blot. ResultsCompared with the normal group， the model group showed polycystic changes in the ovaries， with atrophy of mitochondria in granulosa cells and increased membrane density. Serum levels of testosterone（T）， luteinizing hormone（LH）， and insulin were significantly increased（P<0.01）. The levels of ROS， MDA， 4-hydroxynonenal（4-HNE）， and Fe²⁺ in ovarian tissues were significantly elevated（P<0.01）， while adenosine triphosphate（ATP）， GSH， and reduced nicotinamide adenine dinucleotide phosphate （NADPH） levels were significantly decreased（P<0.01）. The phosphorylation levels of AMPK and acetyl-CoA carboxylase （ACC）， as well as the protein expressions of SLC7A11， GPX4， and ferroptosis suppressor protein 1（FSP1） were significantly downregulated（P<0.01）， whereas the expressions of FASN， ACSL4， and nuclear receptor coactivator 4（NCOA4） were significantly upregulated（P<0.01）. Compared with the model group， MBXT intervention at various doses improved the above pathological changes and biochemical indicators in a dose-dependent manner， with the high-dose group showing the most significant effect（P<0.01）. Compared with the MBXT high-dose group， the high-dose of MBXT combined with ferroptosis inducer Erastin group restored ovarian ferroptosis characteristics in rats， with increased ROS and lipid peroxidation products， and altered expressions of key proteins（P<0.05， P<0.01）. ConclusionMBXT can effectively improve ovarian function and metabolic disorders in PCOS-IR rats. Its mechanism may be related to activating the AMPK/ACC signaling pathway， downregulating FASN and ACSL4 to reduce lipid peroxidation substrates， and restoring glucose-6-phosphate dehydrogenase/phosphoglycerate dehydrogenase（G6PD/PHGDH） metabolic flux to enhance the GPX4/FSP1 antioxidant defense system， thereby inhibiting ferroptosis in ovarian granulosa cells.

Polycystic ovary syndrome （PCOS） with insulin resistance （IR） represents a common complex endocrine-metabolic disorder in reproductive-aged women，posing substantial threats to their reproductive and long-term health. The networked characteristics of pathological mechanisms pose severe challenges in the diagnosis and treatment modes with a single medical system. Western medicine identifies it as a pathological axis with the core of "IR-hyperinsulinemia-hyperandrogenemia" and offers effective symptomatic treatments，it is often faced with limitations such as insufficient overall regulation and drug side effects. Traditional Chinese medicine is characterized by a holistic concept centered on the dysfunction of "kidney-spleen-liver" and syndrome differentiation and treatment，yet its microscopic action mechanisms remain to be fully elucidated. An integrative diagnosis and treatment mode of traditional Chinese medicine and western medicine provides a vital approach to overcoming these challenges. This review systematically sorted out the understanding of polycystic ovary syndrome with insulin resistance （PCOS-IR） pathogenesis from both Chinese and western medicine perspectives. An explainable mapping relation potentially existing between the molecular pathological networks defined by western medicine and the macroscopic dialectical system of traditional Chinese medicine was hypothesized. The synergistic potentiation mechanisms of integrative strategies of traditional Chinese medicine and western medicine were analyzed，with a focus on clarifying the recent progress of integrative diagnosis and treatment strategies. The paper aims to provide a more comprehensive and profound reference for research and clinical practice in this field，advance the further development of the prevention and treatment of PCOS-IR with integrated traditional Chinese medicine and western medicine，and explore the establishment of a more precise，individualized diagnosis and treatment system to make optimal clinical decisions.

ObjectiveTo systematically investigate the identification characteristics of medicinal amber， elucidate its microscopic features， crystal structural properties， and elemental composition， and thereby provide a scientific foundation for quality control and authenticity verification. MethodsThirty-nine batches of amber samples were collected and analyzed through integrated techniques including morphological analysis， microscopic identification， powder X-ray diffraction （XRD）， Raman spectroscopy， Fourier-transform infrared （FTIR） spectroscopy， and inductively coupled plasma mass spectrometry （ICP-MS） to evaluate their morphological attributes， phase composition， molecular vibrational modes， and trace element profiles. Among them， the XRD experiment used Cu Kα radiation （λ=1.540 6 Å）， with a scanning angle range of 10° to 70° （2θ） and a step size of 0.02°， the Raman spectroscopy experiment employed a 785 nm laser， with a spectral measurement range of 3 400 to 50 cm^-1， a laser power of 300 mW， a laser intensity of 30%， and a scanning time of 100 to 1 000 ms， the infrared spectroscopy experiment used a carbon-sulfur lamp， with a scanning range of 4 000 to 500 cm^-1， a resolution of 4 cm^-1， and 3 scans， the ICP-MS experiment utilized frequency power of 1.2 kW， a double-pass cyclonic spray chamber， a sample introduction system flow rate of 0.7-1.0 L·min^-1， and an auxiliary gas flow of 0.2 L·min^-1. ResultsUnder orthogonal polarized light microscopy， medicinal amber exhibited an isotropic homogeneous structure， with partial samples containing inorganic impurities such as AsS and SiO₂. FTIR spectra revealed characteristic absorption peaks at 2 932-2 939 cm^-1 （C-H stretching vibrations）， 1 705-1 728 cm^-1 （C=O stretching vibrations）， and 880-887 cm^-1 （C=C deformation vibrations）， confirming the oxidative polymerization of terpenoid resin. Raman spectroscopy further identified distinctive peaks at 2 925 cm^-1， 2 870 cm^-1 （saturated C-H stretching）， and 1 648 cm^-1 （C=C stretching）， consistent with the structural features of oxidized-polymerized resin. ICP-MS analysis demonstrated that S， Al， Si， Fe， Na， and Ca were the predominant trace elements in medicinal amber. ConclusionThis study comprehensively evaluated medicinal amber's morphological attributes， phase composition， molecular vibrational modes， and trace elements through multimodal analytical techniques. The findings establish data support for establishing quality standards for medicinal amber and distinguishing it from synthetic resin imitations.

ObjectiveTo systematically investigate the identification characteristics of medicinal amber， elucidate its microscopic features， crystal structural properties， and elemental composition， and thereby provide a scientific foundation for quality control and authenticity verification. MethodsThirty-nine batches of amber samples were collected and analyzed through integrated techniques including morphological analysis， microscopic identification， powder X-ray diffraction （XRD）， Raman spectroscopy， Fourier-transform infrared （FTIR） spectroscopy， and inductively coupled plasma mass spectrometry （ICP-MS） to evaluate their morphological attributes， phase composition， molecular vibrational modes， and trace element profiles. Among them， the XRD experiment used Cu Kα radiation （λ=1.540 6 Å）， with a scanning angle range of 10° to 70° （2θ） and a step size of 0.02°， the Raman spectroscopy experiment employed a 785 nm laser， with a spectral measurement range of 3 400 to 50 cm^-1， a laser power of 300 mW， a laser intensity of 30%， and a scanning time of 100 to 1 000 ms， the infrared spectroscopy experiment used a carbon-sulfur lamp， with a scanning range of 4 000 to 500 cm^-1， a resolution of 4 cm^-1， and 3 scans， the ICP-MS experiment utilized frequency power of 1.2 kW， a double-pass cyclonic spray chamber， a sample introduction system flow rate of 0.7-1.0 L·min^-1， and an auxiliary gas flow of 0.2 L·min^-1. ResultsUnder orthogonal polarized light microscopy， medicinal amber exhibited an isotropic homogeneous structure， with partial samples containing inorganic impurities such as AsS and SiO₂. FTIR spectra revealed characteristic absorption peaks at 2 932-2 939 cm^-1 （C-H stretching vibrations）， 1 705-1 728 cm^-1 （C=O stretching vibrations）， and 880-887 cm^-1 （C=C deformation vibrations）， confirming the oxidative polymerization of terpenoid resin. Raman spectroscopy further identified distinctive peaks at 2 925 cm^-1， 2 870 cm^-1 （saturated C-H stretching）， and 1 648 cm^-1 （C=C stretching）， consistent with the structural features of oxidized-polymerized resin. ICP-MS analysis demonstrated that S， Al， Si， Fe， Na， and Ca were the predominant trace elements in medicinal amber. ConclusionThis study comprehensively evaluated medicinal amber's morphological attributes， phase composition， molecular vibrational modes， and trace elements through multimodal analytical techniques. The findings establish data support for establishing quality standards for medicinal amber and distinguishing it from synthetic resin imitations.