1.Exploration of the Application of Fengfu (GV 16) Acupoint in BIAN Que Heart Book (《扁鹊心书》)
Yawei ZHAO ; Haoying LI ; Lintong WEN ; Hefei WANG ; Wei WANG ; Hongyu WU ; Shijiang SUN
Journal of Traditional Chinese Medicine 2025;66(1):98-101
By examining the records related to the Fengfu (GV 16) acupoint in BIAN Que Heart Book (《扁鹊心书》) compiled by the Song Dynasty physician DOU Cai, this study analyzed various aspects, including the differentiation of conditions treated with Fengfu (GV 16) acupoint, the theoretical foundation for selection of Fengfu (GV 16) acupoint, the application of needling manipulation, and the sensation of obtaining qi during acupuncture. The findings suggest that DOU Cai's approach to utilizing Fengfu (GV 16) acupoint differs from traditional methods, particularly emphasizing the effectiveness of achieving a sensation of heat and numbness. His unique techniques include transverse insertion at Fengfu (GV 16) acupoint and penetrated insertion to Fengchi (GB 20) and Yifeng (TE 17) acupoints. The records of Fengfu (GV 16) acupoint in BIAN Que Heart Book provide a valuable reference for its modern clinical application and further development.
2.Clinical Efficacy of Gandou Fumu Granules in Intervention of Liver Fibrosis in WD Patients with Phlegm and Blood Stasis Syndrome and Effect on Cuproptosis-related Indicators
Fei WANG ; Zhenzhen JIANG ; Yimin CHEN ; Zhuang TAO ; Meixia WANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(4):174-181
ObjectiveTo observe the clinical efficacy of Gandou Fumu Granules (GDFMG) combined with sodium dimercaptosulphonate (DMPS) on liver fibrosis in Wilson disease (WD) patients with the syndrome of phlegm and blood stasis, evaluate its effect on cuproptosis-related indicators, and explore the possible mechanisms of cuproptosis in WD-related liver fibrosis. MethodsSixty WD patients diagnosed with the syndrome of phlegm and blood stasis between January 2023 and December 2023 were randomly divided into a control group and an observation group, with 30 patients in each group. The control group received the copper chelator DMPS for the first 6 days, followed by calcium gluconate injection for the next 2 days, completing an 8-day treatment cycle. The observation group received GDFMG in addition to the treatment regimen of the control group, with both groups treated for 21 cycles. A Beckman fully automated biochemical analyzer was used to detect levels of type Ⅳ collagen (CⅣ), hyaluronic acid (HA), laminin (LN), N-terminal propeptide of type Ⅲ procollagen (PⅢ-NP), and serum copper (SCu) before and after treatment in both groups. Enzyme-linked immunosorbent assay (ELISA) was used to measure levels of ferredoxin 1 (FDX1), lipoic acid synthetase (LIAS), and dihydrolipoamide S-acetyltransferase (DLAT). Atomic absorption spectroscopy measured 24-hour urine copper levels before treatment and after the 7, 14, and 21 treatment cycles in both groups. An Fibro Touch (FT) non-invasive liver fibrosis diagnostic device was used to measure liver stiffness (LSM) in both groups before and after treatment. Traditional Chinese medicine syndrome score (TCMSS) was evaluated at the same intervals. Clinical efficacy, adverse events, and safety indicators were also compared. ResultsAfter treatment, levels of CⅣ, HA, LN, and PⅢNP significantly decreased in both groups compared to pre-treatment levels (P<0.01). The observation group showed a more pronounced reduction compared to the control group (P<0.05). There were no statistically significant differences in SCu levels in both groups before and after treatment. After treatment, levels of FDX1,LIAS and DLAT significantly increased in both groups(P<0.01). The observation group showed more notable improvements in these indicators than the control group (P<0.05). After the 7, 14, 21 treatment cycles, 24-hour urine copper levels significantly increased in both groups compared to pre-treatment levels (P<0.01). The observation group had a greater increase in 24-hour urine copper levels than the control group after treatment (P<0.05,P<0.01), and although 24-hour urine copper levels increased after 7 cycles, a gradual decline was observed in subsequent cycles. After treatment, LSM levels significantly decreased in both groups compared to pre-treatment levels (P<0.01), with the observation group showing a greater reduction than the control group (P<0.05). Clinical efficacy was significantly better in the observation group than the control group (P<0.05). No significant differences in the incidence of adverse events or safety indicators were observed between the two groups after treatment. ConclusionGDFMG combined with DMPS can reduce LSM in WD patients with liver fibrosis and the syndrome of phlegm and blood stasis, inhibit cuproptosis, and improve clinical efficacy.
3.The Impairment Attention Capture by Topological Change in Children With Autism Spectrum Disorder
Hui-Lin XU ; Huan-Jun XI ; Tao DUAN ; Jing LI ; Dan-Dan LI ; Kai WANG ; Chun-Yan ZHU
Progress in Biochemistry and Biophysics 2025;52(1):223-232
ObjectiveAutism spectrum disorder (ASD) is a neurodevelopmental condition characterized by difficulties with communication and social interaction, restricted and repetitive behaviors. Previous studies have indicated that individuals with ASD exhibit early and lifelong attention deficits, which are closely related to the core symptoms of ASD. Basic visual attention processes may provide a critical foundation for their social communication and interaction abilities. Therefore, this study explores the behavior of children with ASD in capturing attention to changes in topological properties. MethodsOur study recruited twenty-seven ASD children diagnosed by professional clinicians according to DSM-5 and twenty-eight typically developing (TD) age-matched controls. In an attention capture task, we recorded the saccadic behaviors of children with ASD and TD in response to topological change (TC) and non-topological change (nTC) stimuli. Saccadic reaction time (SRT), visual search time (VS), and first fixation dwell time (FFDT) were used as indicators of attentional bias. Pearson correlation tests between the clinical assessment scales and attentional bias were conducted. ResultsThis study found that TD children had significantly faster SRT (P<0.05) and VS (P<0.05) for the TC stimuli compared to the nTC stimuli, while the children with ASD did not exhibit significant differences in either measure (P>0.05). Additionally, ASD children demonstrated significantly less attention towards the TC targets (measured by FFDT), in comparison to TD children (P<0.05). Furthermore, ASD children exhibited a significant negative linear correlation between their attentional bias (measured by VS) and their scores on the compulsive subscale (P<0.05). ConclusionThe results suggest that children with ASD have difficulty shifting their attention to objects with topological changes during change detection. This atypical attention may affect the child’s cognitive and behavioral development, thereby impacting their social communication and interaction. In sum, our findings indicate that difficulties in attentional capture by TC may be a key feature of ASD.
4.Gandouling Regulates PI3K/Akt/mTOR Autophagy Signaling Pathway via LncRNA H19 for Treatment of Wilson Disease Liver Fibrosis
Xin YIN ; Han WANG ; Daiping HUA ; Lanting SUN ; Yunyun XU ; Wenming YANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(1):131-138
ObjectiveTo investigate the potential mechanisms and pathways through which Gandouling (GDL) exerts its effects in the treatment of liver fibrosis in Wilson disease. MethodsSixty male SD rats were randomly divided into six groups: the normal group, the model group, the GDL low-, medium-, and high-dose groups (0.24, 0.48, 0.96 g·kg-1), and the penicillamine group (90 mg·kg-1), with 10 rats in each group. A copper-loaded Wilson disease rat model was established by gavage administration of 300 mg·kg-1 copper sulfate pentahydrate to all groups except the normal group. Hematoxylin-eosin (HE) staining and Masson staining were used to observe the pathomorphological changes in the liver. Enzyme-linked immunosorbent assay (ELISA) was employed to measure the levels of hyaluronic acid (HA), laminin (LN), procollagen type-Ⅲ peptide (PC-Ⅲ), and collagen type-Ⅳ (C-Ⅳ). Transmission electron microscopy was used to examine the ultrastructure of liver tissues. Real-time quantitative polymerase chain reaction (Real-time PCR) was used to detect the expression levels of liver tissues and serum exosomal long noncoding RNA H19 (LncRNA H19), phosphatidylinositol 3-kinase (PI3K), protein kinase B (Akt), and mammalian target of rapamycin (mTOR). Western blot analysis was performed to assess the expression levels of PI3K, Akt, mTOR, and their phosphorylated forms, as well as autophagy-related proteins Beclin1 and microtubule-associated protein 1 light chain 3B (LC3-Ⅱ/LC3-Ⅰ) in liver tissues. Beclin1 and LC3-Ⅱ fluorescence signal intensity was observed by immunofluorescence. ResultsCompared with the normal group, the model group exhibited inflammatory cell infiltration in hepatocytes, unclear nuclear boundaries with cell cleavage and necrosis, and collagen fiber deposition around confluent areas. The levels of HA, LN, PC-Ⅲ, and C-Ⅳ were significantly elevated (P<0.01). Transmission electron microscopy revealed an increased number of autophagic vesicles, with autophagic lysosomes exhibiting a single-layer membrane structure following degradation of most envelopes. Expression levels of Beclin1 and LC3-Ⅱ/LC3-Ⅰ were significantly increased (P<0.01), and fluorescence signals of Beclin1 and LC3-Ⅱ were markedly enhanced. The protein expression levels of PI3K, Akt, mTOR, p-PI3K, p-Akt, and p-mTOR were reduced (P<0.01), while LncRNA H19 expression was increased (P<0.01), and mRNA expression levels of PI3K, Akt, and mTOR were decreased (P<0.01). After treatment with GDL, the degree of liver fibrosis was significantly improved, with decreased levels of HA, LN, PC-Ⅲ, and C-Ⅳ. The number of autophagic vesicles was significantly reduced, and expression levels of Beclin1 and LC3-Ⅱ/LC3-Ⅰ proteins were lower (P<0.01). The fluorescence signals of Beclin1 and LC3-Ⅱ weakened dose-dependently. The protein levels of PI3K, Akt, mTOR, p-PI3K, p-Akt, and p-mTOR were elevated (P<0.01), while the expression level of LncRNA H19 was reduced (P<0.01). Furthermore, the mRNA expression levels of PI3K, Akt, and mTOR increased (P<0.05, P<0.01). ConclusionGDL may alleviate liver fibrosis and reduce liver injury by regulating the PI3K/Akt/mTOR autophagy signaling pathway via LncRNA H19.
5.Clinical research on corneal epithelium remodeling after small incision lenticule extraction and femtosecond laser-assisted in situ keratomileusis
Yangjing ZHANG ; Liwei MA ; Fan ZHANG ; Chunmei KE ; Ruifu WANG ; Lipye ZU
International Eye Science 2025;25(1):37-41
AIM: To compare the changes in corneal epithelial thickness(CET)after small incision lenticule extraction(SMILE)and femtosecond laser-assisted in situ keratomileusis(FS-LASIK).METHODS: A total of 187 patients(187 eyes)who underwent either SMILE or FS-LASIK at Urumqi Aier Eye Hospital between December 2022 and November 2023 were collected. The patients were divided into SMILE group and FS-LASIK group according to surgical methods. The CET of the patients was measured by optical coherence tomography(OCT)system before surgery and at 1 wk, 1, 3, and 6 mo postoperatively.RESULTS: Changes in corneal epithelial thickness(△CET)in the central, paracentral, and mid-peripheral regions were compared at 6 mo postoperatively. The SMILE group was characterized by the most significant thickening in the central area and the least thickening in the mid-peripheral area; while the FS-LASIK group was characterized by the most significant thickening in the paracentral area and the least thickening in the mid-peripheral region. At 1 wk, 1, 3, and 6 mo postoperatively, within the 0-7 mm corneal area, the △CET for both the SMILE and FS-LASIK groups was correlated with the preoperative spherical equivalent.CONCLUSION: Within 6 mo postoperatively, both SMILE and FS-LASIK showed a similar trend in epithelial thickening but with distinct characteristics. The change in corneal epithelial thickness for both procedures was positively correlated with the preoperative diopter.
6.Mechanism of Gushining Granules in Attenuating Dexamethasone-induced Apoptosis of Bone Marrow Mesenchymal Stem Cells via Activating PI3K/Akt/Bad Signalling Pathway
Chengyu CHU ; Lei ZHU ; Long LIANG ; Feng WANG ; Xuejian YU ; Wenwu LIANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(5):115-122
ObjectiveTo establish steroid-induced osteonecrosis of the femoral head (SANFH) cell model by using dexamethasone (DEX)-induced bone marrow mesenchymal stem cells (BMSCs) and demonstrate that Gushing Granules (GSNs) exert an improving effect by activating the phosphatidylinositol-3-kinase/protein kinase B/B-lymphoma-2 gene related promoter (PI3K/Akt/Bad) signalling pathway. MethodsFirstly, SD rats were orally administered with drugs at a dose of 0.9 g·kg-1 to prepare GSN-containing serum, and CCK-8 screening was used to determine the optimal dosage and duration of action. Then, BMSCs were cultured and treated with 1×10-6 mol·L-1 DEX, 10% GSN-containing serum, and inhibitor LY294002 of PI3K/Akt signalling pathway for 24 hours to model and group SANFH cells. Cell viability and proliferation were detected by using CCK-8 assay kit and EdU staining kit. Flow cytometry was used to detect cell apoptosis. An alkaline phosphatase (ALP) assay kit was employed to detect ALP expression. In order to detect the PI3K/Akt/Bad signalling pathway and protein and mRNA expression of apoptosis-related proteins such as apoptosis regulatory factors B-cell lymphoma-2 gene (Bcl-2), and Bcl-2-associated X protein (Bax), osteocalcin (OCN), and Collagen Ⅰ, we used Western blot and Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR). ResultsThe CCK-8 assay kit determined that the optimal dosage for GSN-containing serum is 10%, and the duration of action is 48 hours. After modelling and grouping the cells in each group, the detection results showed that the SANFH model group had significantly lower cell viability, cell proliferation, and ALP expression, as well as protein and mRNA expressions of PI3K, Akt, Bad, Bcl-2, OCN, and Collagen I compared to the blank group. The nucleic acid and protein levels of the Bax index and the cell apoptosis rate detected by flow cytometry significantly increased (P<0.05,P<0.01). After treatment with GSN-containing serum, cell viability, cell proliferation, and ALP expression, as well as expressions of PI3K, Akt, Bad, Bcl-2, OCN, and Collagen Ⅰ nucleic acids and proteins were significantly increased, while the nucleic acid and protein levels of the Bax index and the cell apoptosis rate detected by flow cytometry significantly decreased(P<0.05,P<0.01). Compared with the GSN drug-containing serum group, the simultaneous treatment with the inhibitor LY294002 and GSN drug-containing serum reversed the improvement effect of GSN. Specifically, the cell viability, cell proliferation, ALP expression, and the nucleic acid and protein levels of PI3K, Akt, Bad, Bcl-2, OCN, and Collagen Ⅰ were all significantly decreased, while the nucleic acid and protein levels of the Bax index and the cell apoptosis rate detected by flow cytometry were significantly increased (P<0.05, P<0.01). ConclusionGSNs antagonize DEX-induced apoptosis of BMSCs by activating the PI3K/Akt/Bad signalling pathway, providing a scientific theoretical basis for the clinical treatment of SANFH with GSNs.
7.Effect of Huangling Jidu Xizhuo Granules on NLRP3, Intestinal Flora and Short-term Prognosis in Patients with Gout
Dandan TIAN ; Shanping WANG ; Li YANG ; Tingting ZHANG ; Xi CHEN ; Chuanbing HUANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(5):150-156
ObjectiveTo investigate the effects of retention enema with Huangling Jiedu Xiezhuo granules(HJXG) on Nod-like receptor protein 3(NLRP3), intestinal flora, and short-term prognosis in patients with gout. MethodsA total of 60 patients with gout admitted to the hospital from January 2021 to December 2023 were selected and divided into a control group and an observation group according to the random number table method, with 30 cases in each group. The control group was treated with febuxostat, and the observation group was treated with retention enema with HJXG on the basis of the control group. After 14 days of continuous treatment, the clinical efficacy, traditional Chinese medicine (TCM) syndrome score, and visual analogue scale (VAS) pain index of the two groups were compared, and serum creatinine(SCr), blood urea nitrogen(BUN), uric acid(UA), cystatin C(CysC), β2- microglobulin(β2-MG), glomerular filtration rate test(GFR), creatinine clearance rate (Ccr), erythrocyte sedimentation rate(ESR), hypersensitive C-reactive protein,(hs-CRP), interleukin 6(IL-6), interleukin-1β(IL-1β), interleukin-18 (IL-18), NLRP3 inflammasome levels, and the number of intestinal flora were detected in the two groups. The prognosis of patients was followed up within 12 weeks. COX regression analysis was used to analyze the effect of short-term prognosis. ResultsAfter treatment, TCM syndrome scores and VAS pain index in both groups were reduced (P<0.05), and TCM syndrome scores in the observation group were significantly lower than those in the control group. After treatment, ESR, hs-CRP, IL-6, NLRP3, IL-18, and IL-1β were significantly decreased in both groups (P<0.01), and the levels of IL-6, ESR, NLRP3, and IL-18 were significantly improved in the observation group compared with the control group (P<0.05). BUN, SCr, UA, β2-MG, GFR indexes in both groups were significantly lower after treatment, Ccr indexes in both groups were significantly higher after treatment, and the levels of SCr, UA, CysC, and Ccr in the observation group were significantly better than those in the control group (P<0.05). After treatment, the intestinal flora in both groups was improved, and the observation group was significantly improved compared with the control group in terms of Lactobacillus, Proteus, Bacteroides, and Escherichia coli (P<0.05). COX regression analysis showed that retention enema with HJXG could reduce the risk of poor short-term prognosis in patients with gout compared with Western medicine alone. ConclusionThe retention enema with HJXG can improve the curative effect of patients with gout, improve the TCM syndromes, reduce inflammation, and enhance renal function, intestinal flora, and short-term prognosis.
8.Analysis on Dynamic Change of Stir-fried Glycyrrhizae Radix et Rhizoma Quality Based on "Exterior-interior Correlation"
Yue XU ; Zhe JIA ; Yun WANG ; Bing LI ; Deling WU ; Cun ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(5):194-202
ObjectiveIn order to provide a reference for the optimization of preparation process of stir-fried Glycyrrhizae Radix et Rhizoma(sf-GRR), the quality changes during the processing was studied. MethodsGlycyrrhizae Radix et Rhizoma was processed by stir-frying for 17 min, and samples were collected every 1 min during the processing. The appearance color of the samples was determined by visual analysis technology, the moisture and extract of the process samples were detected by the drying method and the hot extraction method of alcohol-soluble extract in the general rules of the 2020 edition of Chinese Pharmacopoeia(part Ⅳ), and the contents of liquiritin apioside, liquiritin, isoliquiritin apioside, isoliquiritin, licoricesaponin G2 and glycyrrhizic acid in the process samples were determined by high performance liquid chromatography(HPLC). Then principal component analysis(PCA), partial least squares-discriminant analysis(PLS-DA) and Spearman correlation analysis were used for clustering, discrimination and correlation analysis of the appearance color, moisture, extract and the contents of six internal components. Based on artificial neural network and random forest algorithm, the prediction model of processing degree of sf-GRR was established. On this basis, based on the five principles of quality marker(Q-Maker), explore the monitoring Q-Maker of sf-GRR. ResultsThe color of Glycyrrhizae Radix et Rhizoma deepened after stir-frying, and the appearance color of the sample changed from light yellow to dark yellow during processing. During the stir-frying process, the moisture content showed a decreasing trend with the extension of processing time, while the extract content showed an increasing trend with the extension of processing time. After stir-frying, the contents of liquiritin apioside, liquiritin and licoricesaponin G2 showed an overall decreasing trend, while the contents of isoliquiritin apioside and isoliquiritin increased, and the content of glycyrrhizic acid increased slightly. The correlation analysis showed that moisture was positively correlated with brightness(L*) and red/green value(a*), and negatively correlated with yellow/blue value(b*) and total color difference(E*ab). Isoliquiritin apioside and isoliquiritin had negative correlation with L* and a*, and positive correlation with b* and E*ab. The processing process of sf-GRR could be divided into two stages of the early stage(0-14 min) and the late stage(15-17 min), and could be divided into three stages of the early stage(0-6 min), the middle stage(7-14 min) and the late stage(15-17 min) by combining the moisture, extract, the contents of 6 components and color values. Based on artificial neural network analysis and random forest algorithm, isoliquiritin apioside, isoliquiritin, liquiritin and glycyrrhizic acid were selected as monitoring markers for sf-GRR. ConclusionBased on the analysis of the exterior-interior indicators of process samples of sf-GRR, this paper ultimately identifies four processing monitoring markers, which can provide a basis for optimizing the processing technology of sf-GRR.
9.Protective Effect of Taohong Siwutang on Cerebral Ischemia-reperfusion Injury Based on A1/A2 Phenotype Transformation of Astrocytes Mediated by JAK2/STAT3 Pathway
Huifang WANG ; Xinru CHEN ; Mengyuan CHEN ; Xian ZHOU ; Lan HAN ; Weidong CHEN ; Zhaojie JI
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(7):25-34
ObjectiveTo investigate whether the effect of Taohong Siwutang on cerebral ischemia-reperfusion (CIRI) injury in rats is related to the regulation of astrocyte polarization and explore the related mechanism. MethodsEighty-four male SD rats were randomly assigned to the following groups: A sham operation group, a model group, Taohong Siwutang treatment groups (low dose, medium dose, and high dose), ligustrazine phosphate tablet (LPT) group, and AG490 group. All groups, except for the sham operation group, underwent middle cerebral artery occlusion/reperfusion (MCAO/R) modeling and were treated for seven days. The neurological impairment was evaluated using the Longa score. The volume of cerebral infarction was assessed through 2,3,5-triphenyltetrazolium chloride (TTC) staining. Real-time fluorescent quantitative polymerase chain reaction (Real-time PCR) and Western blot analyses were performed to analyze the mRNA and protein expression levels of cortical complement 3 (C3), S100 calcium-binding protein A10 (S100A10), Janus kinase 2 (JAK2), and signal transducer and activator of transcription 3 (STAT3). Additionally, protein expression levels of vascular endothelial growth factor-A (VEGF-A) were assessed, and the mRNA expression levels of inflammatory factors, including interleukin-6 (IL-6), interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α), were evaluated. Glial fibrillary acidic protein (GFAP) and C3, S100A10 and Co-localization was detected via immunofluorescence double staining. Lastly, VEGF expression levels were measured using enzyme-linked immunosorbent assay (ELISA). ResultsCompared with the sham operation group, the model group showed a significant increase in cerebral infarction volume and neurological impairment (P<0.01). C3 protein levels were elevated, while S100A10 levels were decreased. Pathway-related markers were significantly upregulated (P<0.05, P<0.01), and VEGF-A protein levels were significantly reduced (P<0.01). The mRNA expression of inflammatory factors was significantly upregulated (P<0.01). Co-localization analysis showed significantly increased GFAP and C3 fluorescence intensity (P<0.01) and greatly decreased GFAP and S100A10 fluorescence intensity (P<0.01). Additionally, VEGF content was significantly elevated (P<0.01). Compared with the model group, medium- and high-dose Taohong Siwutang and LPT groups exhibited a significant reduction in cerebral infarction volume and neurological impairment (P<0.01). Groups treated with low, medium, and high doses of Taohong Siwutang and LPT group exhibited a decrease in C3 protein expression levels and an increase in S100A10 expression levels (P<0.01). In the high-dose Taohong Siwutang and AG490 groups, both protein and mRNA expression of C3 and pathway-related markers were significantly downregulated (P<0.05, P<0.01), while S100A10 expression and VEGF-A protein levels were significantly increased (P<0.01). Additionally, the mRNA expression levels of inflammatory factors were significantly reduced (P<0.01). The co-localization fluorescence intensity of GFAP and C3 significantly decreased (P<0.01), while that of GFAP and S100A10 greatly increased (P<0.01). Furthermore, VEGF content exhibited a marked elevation (P<0.01). ConclusionTaohong Siwutang exerts a protective effect in rats with cerebral CIRI injury. The underlying mechanism is associated with the downregulation of the JAK2/STAT3 signaling pathway, promotion of A2-type astrocyte polarization, reduction of inflammatory factor release, and enhancement of VEGF production.
10.Analysis of Differential Compounds of Poria cocos Medicinal Materials by Integrated Qualitative Strategy Based on UPLC-Q-Orbitrap-MS
Jiayuan WANG ; Xiaohan FAN ; Xiaoxiao WEI ; Rong CAO ; Jin WANG ; Lei WANG ; Fengqing XU ; Shunwang HUANG ; Deling WU ; Hongsu ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(7):148-156
ObjectiveTo establish a rapid analytical method for identifying the differential components in Poria cocos medicinal materials based on ultra performance liquid chromatography-quadrupole-electrostatic field orbital trap high-resolution mass spectrometry(UPLC-Q-Orbitrap-MS), combined with mass defect filtering(MDF) and molecular network integration techniques. MethodsUPLC-Q-Orbitrap-MS was used for MS data acquisition and identification of P. cocos medicinal materials, with the help of MDF for the study of cleavage behavior and structural identification of triterpenoids. According to the similarity of MS/MS fragmentation patterns of each component, global natural product social molecular network(GNPS) was established, and Cytoscape 3.6.1 was used to screen molecular clusters with similar structures and the the structure of main compound classes were identified and confirmed. Multivariate statistical analyses such as principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA) were used to screen the differential components of the five P. cocos medicinal materials with the variable importance in the projection(VIP) value>1 and P<0.05 as the criteria. ResultsA total of 66 compounds were identified by database comparison, 8 compounds were newly identified by MDF, 28 compounds were newly identified by GNPS, and a total of 102 chemical compounds were identified, including 43 triterpenoids, 16 saccharides, 26 amino acids and peptides, 3 nucleosides, and 14 other compounds. Triterpenoids were predominant in Poriae Cutis and wild Fushen, amino acids and peptides were the most abundant in Poria and cultivated Fushen, carbohydrates were the most abundant in Poriae Cutis. Type Ⅰ and Ⅱ triterpenoids had higher amounts in Poria and cultivated Fushen, type Ⅲ triterpenoids were more abundant in Poriae Cutis, all four types of triterpenoids were higher in Fushenmu, and type Ⅰ, Ⅱ, and Ⅳ triterpenoids were higher in wild Fushen. A total of 12 common differential chemical constituents were screened, including serine, guanosine, gallic acid, 2-octenal, maltotriose, trametenolic acid, dehydroeburicoic acid, dehydrotrametenolic acid, poricoic acid A, poricoic acid B, poricoic acid E and G, but the relative contents of them varied significantly among different medicinal materials. ConclusionAmong the five P. cocos medicinal materials, the types of constituents are generally similar, but their relative contents differed significantly among these medicinal materials, especially in the distribution of triterpenoids. The integration of UPLC-Q-Orbitrap-MS, MDF and GNPS can provide a reference for the rapid qualitative analysis of other Chinese medicines.

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