1.Correlation between spinal sagittal imbalance and knee joint parameters detected by whole-body EOS imaging
Feng ZHOU ; Pengfei FU ; Yufan QIAN ; Pingcheng XU ; Jiongjiong GUO ; Lei ZHANG
Chinese Journal of Tissue Engineering Research 2026;30(3):596-603
BACKGROUND:With changing lifestyles and aging,sagittal spinal imbalance has become a common orthopedic issue significantly affecting knee and pelvic function.Understanding the impact of sagittal spinal imbalance and its compensatory mechanisms is crucial for improving the clinical management of chronic pain.OBJECTIVE:To evaluate the alignment of the spine-pelvis-lower extremities using whole-body EOS imaging,analyze the correlation between spinal sagittal imbalance and knee joint parameters,and explore their compensatory mechanisms.METHODS:A total of 71 patients with chronic low back pain or patellofemoral pain who visited Department of Orthopedics,First Affiliated Hospital of Soochow University between January 1,2021 and December 31,2023 were included.Radiographic measurements were performed using whole-body EOS to determine pelvic tilt,pelvic incidence,lumbar lordosis,sagittal vertical axis,global tilt,hip-knee-angle,knee flexion angle,lateral distal femoral angle,and medial proximal tibial angle.Patients were classified into normal group(pelvic incidence-lumbar lordosis<10°),compensated group(10°<pelvic incidence-lumbar lordosis<20°),and decompensated group(pelvic incidence-lumbar lordosis>20°)based on the SRS-Schwab spinal deformity classification according to pelvic incidence-lumbar lordosis difference.The differences in radiographic parameters among the groups were analyzed.The differences in American Knee Society Knee Score and Oswestry Disability Index scores were compared among each group.Patients were divided into chronic low back pain group and non-chronic low back pain group,patellofemoral pain group and non-patellofemoral pain group based on clinical symptoms,and the relationship between radiographic parameter differences and clinical symptoms was analyzed.RESULTS AND CONCLUSION:(1)When pelvic incidence-lumbar lordosis was less than 20°,lateral distal femoral angle and medial proximal tibial angle tended to stabilize.When pelvic incidence-lumbar lordosis was greater than 20°,it showed a linear correlation with lateral distal femoral angle and medial proximal tibial angle,with lateral distal femoral angle increasing and medial proximal tibial angle decreasing with increasing pelvic incidence-lumbar lordosis values.(2)Compared with the normal group,the compensated group had significantly increased pelvic tilt(P<0.01),while knee joint parameters hip-knee-angle and knee flexion angle showed no significant differences;the decompensated group showed significant increases in pelvic tilt(P<0.01),and decreases in hip-knee-angle,and knee flexion angle(P<0.01).Compared with the compensated group,the decompensated group showed a significant decrease in hip-knee-angle(P<0.05),but had no significant differences in pelvic tilt and knee flexion angle.(3)Compared with the non-patellofemoral pain group,patients with patellofemoral pain had significant decreases in spinal lumbar lordosis,lateral distal femoral angle,and medial proximal tibial angle(P<0.05)and a significant increase in pelvic incidence-lumbar lordosis(P<0.05).(4)Patients with low back pain had significant differences in radiographic parameters compared with the non-chronic low back pain group(P<0.05).(5)Compared with the normal group,both the compensated and decompensated groups showed a significant decrease in American Knee Society Knee Score scores and a significant increase in Oswestry Disability Index scores(P<0.05).Compared with the compensated group,the decompensated group showed a significant decrease in American Knee Society Knee Score scores and a significant increase in Oswestry Disability Index scores(P<0.05).(6)Pelvic incidence-lumbar lordosis values increased with age and were higher in females compared with males.(7)This study systematically reveals the spine and lower limbs play an important role in disease progression and clinical symptoms.Associated symptoms low back pain and patellofemoral pain are related to the stability of the spine-pelvis-lower extremity alignment.Furthermore,spinal sagittal imbalance is more severe in elderly and female patients.
2.Correlation between spinal sagittal imbalance and knee joint parameters detected by whole-body EOS imaging
Feng ZHOU ; Pengfei FU ; Yufan QIAN ; Pingcheng XU ; Jiongjiong GUO ; Lei ZHANG
Chinese Journal of Tissue Engineering Research 2026;30(3):596-603
BACKGROUND:With changing lifestyles and aging,sagittal spinal imbalance has become a common orthopedic issue significantly affecting knee and pelvic function.Understanding the impact of sagittal spinal imbalance and its compensatory mechanisms is crucial for improving the clinical management of chronic pain.OBJECTIVE:To evaluate the alignment of the spine-pelvis-lower extremities using whole-body EOS imaging,analyze the correlation between spinal sagittal imbalance and knee joint parameters,and explore their compensatory mechanisms.METHODS:A total of 71 patients with chronic low back pain or patellofemoral pain who visited Department of Orthopedics,First Affiliated Hospital of Soochow University between January 1,2021 and December 31,2023 were included.Radiographic measurements were performed using whole-body EOS to determine pelvic tilt,pelvic incidence,lumbar lordosis,sagittal vertical axis,global tilt,hip-knee-angle,knee flexion angle,lateral distal femoral angle,and medial proximal tibial angle.Patients were classified into normal group(pelvic incidence-lumbar lordosis<10°),compensated group(10°<pelvic incidence-lumbar lordosis<20°),and decompensated group(pelvic incidence-lumbar lordosis>20°)based on the SRS-Schwab spinal deformity classification according to pelvic incidence-lumbar lordosis difference.The differences in radiographic parameters among the groups were analyzed.The differences in American Knee Society Knee Score and Oswestry Disability Index scores were compared among each group.Patients were divided into chronic low back pain group and non-chronic low back pain group,patellofemoral pain group and non-patellofemoral pain group based on clinical symptoms,and the relationship between radiographic parameter differences and clinical symptoms was analyzed.RESULTS AND CONCLUSION:(1)When pelvic incidence-lumbar lordosis was less than 20°,lateral distal femoral angle and medial proximal tibial angle tended to stabilize.When pelvic incidence-lumbar lordosis was greater than 20°,it showed a linear correlation with lateral distal femoral angle and medial proximal tibial angle,with lateral distal femoral angle increasing and medial proximal tibial angle decreasing with increasing pelvic incidence-lumbar lordosis values.(2)Compared with the normal group,the compensated group had significantly increased pelvic tilt(P<0.01),while knee joint parameters hip-knee-angle and knee flexion angle showed no significant differences;the decompensated group showed significant increases in pelvic tilt(P<0.01),and decreases in hip-knee-angle,and knee flexion angle(P<0.01).Compared with the compensated group,the decompensated group showed a significant decrease in hip-knee-angle(P<0.05),but had no significant differences in pelvic tilt and knee flexion angle.(3)Compared with the non-patellofemoral pain group,patients with patellofemoral pain had significant decreases in spinal lumbar lordosis,lateral distal femoral angle,and medial proximal tibial angle(P<0.05)and a significant increase in pelvic incidence-lumbar lordosis(P<0.05).(4)Patients with low back pain had significant differences in radiographic parameters compared with the non-chronic low back pain group(P<0.05).(5)Compared with the normal group,both the compensated and decompensated groups showed a significant decrease in American Knee Society Knee Score scores and a significant increase in Oswestry Disability Index scores(P<0.05).Compared with the compensated group,the decompensated group showed a significant decrease in American Knee Society Knee Score scores and a significant increase in Oswestry Disability Index scores(P<0.05).(6)Pelvic incidence-lumbar lordosis values increased with age and were higher in females compared with males.(7)This study systematically reveals the spine and lower limbs play an important role in disease progression and clinical symptoms.Associated symptoms low back pain and patellofemoral pain are related to the stability of the spine-pelvis-lower extremity alignment.Furthermore,spinal sagittal imbalance is more severe in elderly and female patients.
3.Effect and Mechanism of Xiao Qinglongtang Against Right Ventricular Dysfunction in Rats with Pulmonary Arterial Hypertension Induced by Monocrotaline
Lei QI ; Huifei ZHANG ; Ling GONG ; Jifu HE ; Wenjing CHEN ; Weipin NIU ; Xiao LI ; Yuehua JIANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(4):11-19
ObjectiveThis study aimed to establish a monocrotaline (MCT)-induced pulmonary arterial hypertension (PAH) rat model to systematically evaluate the protective effect of Xiao Qinglongtang (XQLT) on right cardiac function in model rats and further elucidate the underlying regulatory mechanism. MethodsSixty male SD rats were randomly assigned to the normal group, model group, XQLT low-, medium-, and high-dose groups (XQLT-L/M/H), and the beraprost sodium tablet group (BST). Except for the normal group, rats in all other groups were given a single subcutaneous injection of MCT (60 mg·kg-1) to induce PAH. Three weeks after injection, rats in the XQLT-L/M/H groups were administered XQLT intragastrically at 3.07, 6.14, 12.28 g·kg-1·d-1, respectively. Rats in the BST group received beraprost sodium at 12.6 μg·kg-1·d-1, and rats in the model group received an equal volume of saline. All treatments lasted for 3 weeks. Right ventricular systolic pressure (RVSP) was measured by right ventricular catheterization. Cardiac function was assessed by echocardiography. The right ventricle was weighed to calculate the right ventricular hypertrophy index (RVHI). Hematoxylin-eosin (HE) staining, Masson staining, and transmission electron microscopy were used to observe myocardial morphology. Serum metabolomic changes were analyzed using ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Data-independent acquisition (DIA) proteomics was used to detect differentially expressed (DE) proteins in the right ventricle, and Western blot was used to measure the expression of uncoupling protein 3 (UCP3), phosphatidylinositol 3-kinase catalytic subunit p110α (PIK3CA), L1 cell adhesion molecule (L1CAM), and quinone oxidoreductase (CRYZ). UPLC-MS/MS was used to analyze the chemical components of XQLT. ResultsCompared with the normal group, the model group showed significantly increased RVSP and RVHI (P<0.05), along with pathological changes in myocardial morphology. Compared with the model group, all XQLT-treated groups exhibited reductions in RVSP and RVHI as well as significant improvements in cardiac function and myocardial morphology. Among the XQLT groups, XQLT-M showed the most pronounced effects (P<0.05), comparable to the BST group. Serum metabolomics revealed 105 differential metabolites in the XQLT groups versus the model group [variable importance in projection (VIP) >1, P<0.05], including 58 upregulated and 47 downregulated metabolites. KEGG enrichment analysis indicated that XQLT intervention downregulated phenylalanine metabolism (P<0.01) and upregulated unsaturated fatty acid biosynthesis (P<0.05). Proteomics analysis showed that 982 DE proteins were identified in the MCT groups versus the normal group, including 455 upregulated and 527 downregulated proteins (|fold change (FC)| >1.3, P<0.05). Compared with the model group, 237 DE proteins were identified in the XQLT groups, including 124 upregulated and 113 downregulated proteins (|FC| >1.3, P<0.05), with 57 overlapping DE proteins. KEGG enrichment suggested that XQLT mainly modulated pathways related to mineral absorption, ribosomal biogenesis, peroxisomes, glycolysis/gluconeogenesis, spliceosomes, and thyroid hormone signaling. Western blot analysis showed that, compared with the model group, XQLT increased the expression of UCP3, PIK3CA, and L1CAM, while decreasing the expression of CRYZ (P<0.05). ConclusionXQLT exerts a protective effect on right heart function in MCT-induced PAH rats, and its mechanism is associated with maintaining myocardial homeostasis and alleviating right ventricular remodeling.
4.Action Mechanism of Huamoyan Granules in Treatment of Knee Osteoarthritis Based on TRPV1/p38 MAPK Pathway
Jin ZHANG ; Lili YANG ; Canwen ZHENG ; Jing KANG ; Yanlei MA ; Yue SHI ; Lei LI ; Hongxu MENG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(4):79-89
ObjectiveThis paper aims to observe the protective effect of Huamoyan granules on knee osteoarthritis (KOA) and explore whether its protective effect is oriented toward an anti-inflammatory direction by regulation of macrophage polarization, which can effectively inhibit the progression of pathological inflammatory response, reduce the release of inflammatory pain mediators, and downregulate the protein expression level of transient receptor potential vanilloid 1 (TRPV1), so as to provide experimental evidence for its clinical application and investigate its action mechanism. MethodsAfter adaptive feeding, Sprague-Dawley (SD) rats were randomly divided into six groups: sham group, model group, celecoxib group, and high, medium, and low-dose synovitis granule groups (9.6, 4.8, 2.4 g·kg-1). The administration dose of celecoxib capsules was 20 mg·kg-1. There were 10 rats in the sham group and 12 rats in the model group and each administration group. A KOA animal model was established by means of intra-articular injection of sodium iodoacetate into the knee joint. From the 10th day of the experiment, each administration group was given intragastric administration at a dose of 10 mL·kg-1 for 4 weeks. General conditions of rats in each group were assessed daily. The pressure pain threshold (PPT) to mechanical stimulation and joint diameter were recorded. X-ray examination was performed on the right knee joints of rats for imaging analysis. Enzyme linked immunosorbent assay (ELISA) was performed to detect the tumor necrosis factor-α (TNF-α), serum interleukin-1β (IL-1β), and other pro-inflammatory cytokines in rat serum samples, as well as the expression levels of neurogenic inflammatory mediators such as nerve growth factor (NGF) and calcitonin gene-related peptide (CGRP). Histopathological changes in the knee joint synovial tissues were examined by hematoxylineosin (HE) staining. Safranin O-fast green staining was performed to observe and evaluate the degree of knee cartilage lesions. Western blot was employed to quantitatively analyze TRPV1, p38 mitogen-activated protein kinase (p38 MAPK), and phosphorylated (p)-p38 MAPK in rat knee synovial tissues. Immunofluorescence (IF) was used to measure and assess M1/M2 macrophage polarization. ResultsCompared with those in the sham group, the circumference and joint diameter of the right knee were markedly enlarged in the model group (P<0.01), while PPTs of rats showed a significant reduction (P<0.01). The contents of IL-1β, TNF-α, CGRP, and NGF in rats' serum were significantly elevated (P<0.01), and the synovial Krenn score was increased (P<0.01). The Mankin score of cartilage tissue was increased (P<0.01), and the protein expressions of TRPV1 and p-p38 MAPK/p38 MAPK were significantly upregulated (P<0.01). The experimental intervention significantly reduced the proportion of pro-inflammatory M1 macrophages in the total macrophage population (P<0.01), and the percentage of M2 macrophages was decreased (P<0.01). The M1/M2 macrophage ratio was significantly elevated (P<0.01). Knee joint diameters of all dose groups of Huamoyan granules and the celecoxib group were reduced (P<0.01) compared with those of the model group, and the PPT recovery speeds in the high and medium-dose groups of Huamoyan granules were more obvious (P<0.05). The contents of IL-1β, CGRP, and NGF in the rats' serum in all administration groups were significantly reduced (P<0.05, P<0.01), and the content of TNF-α in rats' serum was significantly reduced (P<0.01). All dose groups of Huamoyan granules demonstrated significant reductions in both synovial Krenn score (P<0.05, P<0.01) and protein expression of TRPV1 and p-p38 MAPK/p38 MAPK in rats' synovial tissues (P<0.01). The percentage of M1 macrophages in the synovial tissues of the celecoxib group and all dose groups of Huamoyan granules was decreased (P<0.01). The percentage of M2 macrophages was increased (P<0.05), and the M1/M2 ratio was decreased (P<0.01). ConclusionHuamoyan granules can alleviate the inflammatory response of KOA, reduce the release of inflammatory pain mediators, and downregulate TRPV1 protein expression by regulating macrophage polarization. Its mechanism may be related to the TRPV1/p38 MAPK signaling pathway, thereby achieving the effect of improving peripheral pain hypersensitivity in KOA.
5.Development and evaluation of classification system for drug-related problems in China
Shuang ZOU ; Tingting LU ; Lei BAO ; Yun LIAO ; Ling LI ; Ping ZHANG
China Pharmacy 2026;37(3):371-376
OBJECTIVE To establish a Chinese drug-related problem (DRP) classification system applicable to pharmacist-led pharmaceutical care in China, providing pharmacists with an effective and practical tool for pharmaceutical care. METHODS A multi-stage process was employed to construct the DRP classification system, including literature review and analysis, comparison of existing classification systems, refinement of classification items and framework development, two rounds of standard case validation, expert discussion, and system revision. The Fleiss′ kappa test was used to calculate the consistency coefficient κ, assessing the reliability of pharmacists participating in evaluating the classification system. An electronic questionnaire comprising six items was employed to evaluate the system’s applicability. RESULTS The constructed Chinese DRP classification system comprised six sections [problem(including potential problems), DRP evaluation, cause (including possible causes of potential problems), intervention, acceptance of intervention and DRP status], with 24 primary codes and 96 secondary codes. In the first round of case validation, κ values exceeded 0.4 for all sections except “intervention” and “DRP status”. In the second round, κ values exceeded 0.4 for all sections. In the applicability evaluation of the classification system, positive ratings (“strongly agree” or “agree”) exceeded 85% for all items. Specifically, positive ratings for“the classification system can provide appropriate category selection”,“ the classification system is comprehensive”,“ the classification system is convenient to use” and “the classification system is highly satisfactory” exceeded 92%. CONCLUSIONS The Chinese DRP classification system developed demonstrates both high reliability and applicability, providing an effective and practical classification tool for pharmacists in China to conduct pharmaceutical care.
6.Early warning of influenza epidemic based on CUSUM and EWMA models in Daxing District, Beijing
Hong LEI ; Qiuling LI ; Qi LIU ; Meichen LIU ; Enhuan DU ; Jinfeng TANG ; Zhiping LI ; Yadi GAN ; Lijie ZHANG
Journal of Public Health and Preventive Medicine 2026;37(1):13-17
Objective To explore the effectiveness of the cumulative sum (CUSUM) and the exponentially weighted moving average (EWMA) for early warning of influenza epidemic using two datasets of reported influenza cases and influenza-like illness (ILI) cases. Methods Using the reported cases of influenza and ILI in Daxing District, Beijing, from week 23 of 2018 to week 22 of 2024 as data sets, the CUSUM and EWMA models were established, respectively. The positive rate of influenza etiology was used as the “gold standard”, and the Youden index was used as the evaluation index to compare the early warning effect of the two models under different data sets and different parameters. Results In CUSUM, the optimal Youden indices of the reported influenza cases set and the ILI cases set were 0.751 and 0.635, respectively. In EWMA, the optimal Youden indices of the reported influenza cases set and the ILI cases set were 0.544 and 0.464, respectively. The optimal EWMA and CUSUM models could both issue early warning signals in advance of the “gold standard”. Conclusion In the influenza epidemic early warning in Daxing District, Beijing, the CUSUM model established with the reported cases of influenza can achieve good early warning effects, but the model parameters need to be dynamically adjusted according to the local epidemic characteristics.
7.Mechanism of NAFLD-associated Intestinal Barrier Damage and Traditional Chinese Medicine Intervention Strategies Based on "Turbid Pathogenic Factors Entering the Blood" Theory
Haoyang QIN ; Lei LUO ; Mengge LI ; Xueqian KONG ; Fanghua ZHANG ; Zhongqin DANG ; Zhibo DANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(1):277-287
Intestinal barrier damage is a prominent feature of non-alcoholic fatty liver disease (NAFLD) and serves as a critical factor driving the progression from simple fatty liver to non-alcoholic steatohepatitis (NASH), fibrosis, and cirrhosis. The "turbid pathogenic factors entering the blood" theory integrates classical traditional Chinese medicine (TCM) principles with contemporary disease evolution trends and research findings. It posits that endogenous turbid pathogenic factors within the body infiltrate the blood vessels, leading to impure and viscous blood quality, thereby triggering various diseases. Based on this theory, this article elucidated the pathogenic mechanism of NAFLD-associated intestinal barrier damage. It argued that in NAFLD, the liver loses its dredging function, and the spleen becomes obstructed and dysfunctional. Moreover, essential nutrients fail to be properly transformed, resulting in the internal generation of turbid pathogenic factors. This subsequently initiates a series of pathological changes, namely, "infiltration of phlegm-turbidity into the blood, eroding the intestinal mucosa", "infiltration of glucose-turbidity into the blood, macerating and eroding the intestinal mucosa", "infiltration of heat-turbidity into the blood, scorching and eroding the intestinal mucosa", and "infiltration of stasis-turbidity into the blood, stagnating and eroding the intestinal mucosa", ultimately causing intestinal barrier damage. Furthermore, guided by the "turbid pathogenic factors entering the blood" theory, this article explored TCM intervention strategies: employing medicinals targeting the liver meridian to address the root cause and reduce the generation and deposition of turbid pathogenic factors in the liver, administering blood-system medicinals to clear the blood and purge turbidity, thereby intercepting the progression of the disease mechanism, and applying tonifying medicinals to bolster healthy Qi and defend against turbid invasion, allowing the damaged intestinal mucosa to gradually heal. This article presented novel theoretical and medicinal perspectives for analyzing NAFLD-associated intestinal barrier damage based on the "turbid pathogenic factors entering the blood" theory, aiming to provide new entry points and broader horizons for related research and clinical practice.
8.Microscopic Mechanism of Ulcerative Colitis and New Ideas on Medicine Management Based on Theory of Mutual Interference Between Lucidity and Turbidity
Yuying XU ; Changpu ZHAO ; Lei LUO ; Renwu CHEN ; Zishun LI ; Meiling LI ; Rongzhi LI ; Yu ZHANG ; Guangjie SHU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(1):288-299
The chapter Zhouyu in Guoyu says "Qi of the heaven and the earth moves without losing its order." With lucidity ascending and turbidity descending, Qi moves in a normal state, and Yin and Yang consolidate the foundation of the body. The mutual interference between lucidity and turbidity leads to the disorder of Qi movement, thus causing diseases. It is a pathological state of disorder between ascending and descending, as well as between entering and exiting, gradually evolving into a state of turbidity affecting lucidity and transforming into pathogen, which can be used to interpret and analyze the core of disease pathogenesis. The theory of lucidity and turbidity is connected with the harmony of nutrient and defensive aspects, Qi circulation, and sweat pore associating with Qi movement, and it has common implications with immune responses and nutrient metabolism system, intestinal mucosal barrier function, and mitochondrial energy synthesis. Modern studies have shown that intestinal flora imbalance, bile acid receptor inactivation, macrophage polarization imbalance, epithelial-mesenchymal transition, ferroptosis and other related microscopic pathological mechanisms are involved in the development and progression of ulcerative colitis. By delving into the common meaning of the classic theory of mutual interference between lucidity and turbidity in traditional Chinese medicine and modern medical pathological mechanisms, this paper summarizes the correspondence between the micropathological mechanism and the theory of mutual interference between lucidity and turbidity in the regulation and mamagement of ulcerative colitis. The combined use of sweet and warm medicinal materials consolidates the middle Qi and activates Qi circulation, thus ascending lucidity and descending turbidity. The combined use of pungent medicinal materials for dispersing and bitter medicinal materials for descending simultaneously raises warm and clear Qi. Wind-extinguishing medicinal materials facilitate the ascending of Qi and the opening of sweat pores. Accordingly, turbidity descends and lucidity ascends. The prescriptions incorporating these medication principles are in agreement with the therapeutic approach of following the normal flow of lucidity and turbidity. This paper clarifies the scientific connotation and micropathologic mechanism of ulcerative colitis from the perspective of mutual interference between lucidity and turbidity, providing new theories and prescriptions for the clinical diagnosis, treatment, and prevention of ulcerative colitis.
9.Exploration on Scientific Connotation of "Removing Calyx" of Inulae Flos Based on Processing Experience of Old Medicine Workers
Qiao ZHOU ; Zhenni QU ; Menghan LIU ; Lei ZHANG ; Benzheng SU ; Yanpeng DAI ; Dianhua SHI
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(7):260-266
ObjectiveTo conduct a comparative quality analysis of Inulae Flos, fuzz of Inulae Flos and calyx of Inulae Flos, elucidating the scientific connotation of the "removing calyx" process in the traditional processing of Inulae Flos. MethodsInulae Flos decoction pieces were collected, and the fuzz and calyx of Inulae Flos were prepared according to the experiences of old medicine workers. Subsequently, according to the methods under the "Inulae Flos" item in the 2025 edition of the Pharmacopoeia of the People's Republic of China, the appearance characteristics and thin-layer chromatography(TLC) identification of these samples were tested, and the moisture content, total ash content, extract content were also measured. The characteristic fingerprint patterns of Inulae Flos and fuzz of Inulae Flos were established by high-performance liquid chromatography(HPLC), followed by similarity evaluation, principal component analysis(PCA), and partial least squares-discriminant analysis(PLS-DA). The contents of cryptochlorogenic acid, caffeic acid, 1,3-O-dicaffeoylqunic acid, 1,5-O-dicaffeoylqunic acid, and 1-O-acetyl britannilactone were determined to compare the quality differences of Inulae Flos, fuzz of Inulae Flos, and calyx of Inulae Flos. ResultsThe moisture content of Inulae Flos, fuzz of Inulae Flos, and calyx of Inulae Flos was all<10%. The determination results of total ash content were as follows:Calyx of Inulae Flos>Inulae Flos>fuzz of Inulae Flos, and the determination results of alcohol-soluble extract content were as follows:Fuzz of Inulae Flos>Inulae Flos>calyx of Inulae Flos. HPLC fingerprint patterns of Inulae Flos and fuzz of Inulae Flos were established, and 22 common peaks were identified. The similarity analysis and PCA showed that the overall quality of Inulae Flos and fuzz of Inulae Flos was similar, while the overall quality of calyx of Inulae Flos differed significantly from that of Inulae Flos and fuzz of Inulae Flos. PLS-DA results showed that Inulae Flos, fuzz of Inulae Flos, and calyx of Inulae Flos clustered into distinct groups, indicating significant differences among them. Cryptochlorogenic acid and caffeic acid had relatively high contents in calyx of Inulae Flos, the contents of 1,3-O-dicaffeoylqunic acid and 1,5-O-dicaffeoylqunic acid in Inulae Flos and fuzz of Inulae Flos were higher than those in calyx of Inulae Flos. The order of 1-O-acetyl britannilactone content was determined as follows:fuzz of Inulae Flos>Inulae Flos>calyx of Inulae Flos. ConclusionThe scientific nature of "Removing Calyx" process in the cleansing of Inulae Flos by old medicine workers is demonstrated by the resulting fuzz of Inulae Flos decoction pieces exhibiting enhanced cleanliness and higher content of the index component 1-O-acetyl britannilactone. This study provides a reference basis for further improving and enhancing the processing method and quality control standards of Inulae Flos.
10.Liuwei Dihuangwan Promote Mitophagy to Modulate Neuroinflammation and Behavioral Impairments in Rat Model of Autism Spectrum Disorder (ASD)
Pengjue HUANG ; Mingyue JIANG ; Ji WU ; Niya YIN ; Lei OUYANG ; Qinquan ZHU ; Di ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(2):52-60
ObjectiveTo observe the effect of Liuwei Dihuangwan on behavioral impairments in the rat model of autism spectrum disorder (ASD) and explore the mechanism of action. MethodsTwelve SD pregnant rats were intraperitoneally injected with valproic acid (VPA) (10 rats) or normal saline (2 rats), and male offspring were selected to establish the model of ASD and the control rats. Rats were randomly assigned into model, low-dose (0.75 g·kg-1) and high-dose (1.5 g·kg-1) Liuwei Dihuangwan, vitamin D (positive drug, 3.7×10-5 g·kg-1), and blank groups. Each group was administrated with the corresponding concentration of drugs or the same volume of normal saline by gavage for 2 weeks. After the intervention, the three-chamber social test was conducted to evaluate social interaction and social preference. The open field test was carried out to observe spontaneous behavior and anxiety state. Hematoxylin-eosin staining (HE) was used to observe the pathological changes of the prefrontal tissue. Transmission electron microscopy was employed to observe the ultrastructure of mitochondria in prefrontal neurons. Immunofluorescence was used to detect the expression of ionized calcium-binding adapter molecule-1 (Iba-1) in the prefrontal tissue. Enzyme-linked immunosorbent assay (ELISA) was adopted to measure the levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6). Western blot was employed to assess the expression differences of phosphorylated adenosine monophosphate-activated protein kinase (p-AMPK), adenosine monophosphate-activated protein kinase (AMPK), phosphorylated Unc-51-like autophagy-activating kinase 1 (p-ULK1), Unc-51-like autophagy-activating kinase 1 (ULK1), and FUN14 domain-containing protein 1 (FUNDC1). ResultsCompared with the blank group, the model group spent less time sniffing stranger 1 and stranger 2 in the three-chamber social test (P<0.01) and showed reductions in the total distance traveled, average speed, distance traveled in the central area, and time spent in the central area in the open field test (P<0.01). In addition, the model group showed extensive apoptosis of neurons, with shrunken nuclei and red-stained cytoplasm, and extensive necrosis of neurons in the prefrontal tissue, mitochondrial swelling, decreased matrix density, disrupted cristae, and autophagic lysosomes in neurons, increases in the rate of Iba-1 positive cells in the prefrontal area (P<0.01) and the levels of TNF-α and IL-6 (P<0.01), and down-regulation in the expression of p-AMPK/AMPK, p-ULK1/ULK1, and FUNDC1 (P<0.01). Compared with the model group, low-dose and high-dose Liuwei Dihuangwan and the vitamin D prolonged the time spent sniffing stranger 1 and stranger 2 in the three-chamber social test (P<0.05, P<0.01), increased the total distance traveled, average speed, distance traveled in the central area, and time spent in the central area in the open field test (P<0.05, P<0.01), restored the morphology of neurons in the prefrontal tissue, decreased the number of apoptotic cells, alleviated the swelling of mitochondria in neurons, increased the matrix density, mitigated the fragmentation and disorder of cristae, and increased the number of autophagosomes. Moreover, the drugs decreased the rate of Iba-1 positive cells in the prefrontal area (P<0.01), lowered the levels of TNF-α and IL-6 (P<0.01), and up-regulated the expression of p-AMPK/AMPK, p-ULK1/ULK1, and FUNDC1 (P<0.01). ConclusionLiuwei Dihuangwan ameliorate autism-like behaviors and reduce neuronal apoptosis and neuroinflammatory damage in the rat model of ASD by promoting mitophagy mediated by the AMPK/ULK1/FUNDC1 pathway.


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