1.Differentiation and Treatment of Attention Deficit Hyperactivity Disorder from the Perspective of Deficiency,Stasis and Stagnation
Kangning ZHOU ; Meifang LI ; Yurou YAN ; Yuan LI ; Xi CHEN ; Wei LI ; Hongsheng YANG ; Junhong WANG
Journal of Traditional Chinese Medicine 2026;67(10):1111-1114
The core pathogenesis of attention deficit hyperactivity disorder (ADHD) lies in deficiency, stasis and stagnation. Deficiency arises from kidney essence depletion and spleen dysfunction in transportation and transformation, leading to inadequate nourishment of the marrow sea. Stasis caused by qi deficiency leads to obstruction in channels and collaterals, resulting in obstructed marrow transport. Stagnation is associated with the excess of the five minds transforming into fire, which scorches the brain orifices and leads to loss of control over marrow utilisation. Based on this, a "supplementation-unblocking-regulation" therapeutic approach is proposed. For deficiency, the focus is on supplementing kidney and fortifying spleen, and replenishing the marrow sea. For stasis, the priority is to unblock and open the orifices, and clear the marrow channels. For stagnation, the core is to clear fire and contain the mind, regulate and restore vital activity. In clinical practice, it is necessary to identify the primary and secondary pathogenic mechanisms and apply dynamic, combined treatment, integrating Chinese herbal medicine, acupuncture, and guiding exercises throughout the process, aiming to provide a reference for the diagnosis and treatment of ADHD with traditional Chinese medical.
2.A panel study on association of short-term air pollution exposure and peripheral blood microparticles in healthy adults
Bin ZHANG ; Xinghou HE ; Jiahui LIU ; Xuyang SHAN ; Yan FANG ; Huiying XU ; Erlu ZHAO ; Shengcong LIU ; Hongbing XU ; Jianping LI ; Wei HUANG
Journal of Environmental and Occupational Medicine 2026;43(1):1-7
Background Microparticles (MPs) are one of the main medium of inflammatory reaction with an important role in atherosclerotic progression. Studies on association of air pollution exposure and levels of peripheral blood MPs are limited among human. Objective To evaluate the effects of short-term exposure to air pollution on levels of peripheral blood MPs. Method A panel of 73 healthy adults was followed with 4 repeated follow-ups in Beijing, China, from November 2014 to January 2016. During each visit, we collected questionnaire information, fasting venous blood, urine, and exposures to fine particulate matter (PM2.5), black carbon, nitric oxide, nitrogen dioxide, nitrogen oxide, sulfur dioxide, carbon monoxide, and ozone. We used linear mixed-effect models to analyze associations of air pollution exposure with levels of total MPs (TMPs) and MPs derived from various cells. Stratified analysis was conducted by levels of C-reactive protein (CRP) and malondialdehyde (MDA). Results The results showed significant associations between air pollution exposure and peripheral blood TMPs at 2 h-6 d prior to the follow-ups (P<0.05), while no statistical associations were found for MPs derived from different cell types. Significant increases in TMPs of 7.8% (95%CI: 0.7%, 15.3%) and 14.3% (95%CI: 2.8%, 27.2%) were observed with each interquartile range (IQR) increase in PM2.5 (IQR=64.9 μg·m−3) at prior 18 h and NO (IQR=40.5 μg·m−3) at prior 48 h. Among participants with low levels of CRP and MDA, significantly positive associations were observed between air pollution exposure and levels of TMPs (P<0.05). Conclusion Short-term exposure to air pollution is significantly associated with increased levels of circulating MPs in healthy adults, and in people with lower systemic inflammation, peripheral blood MPs levels are more easily affected after exposure to air pollutants.
3.Development and Validation of a High-Performance Liquid Chromatography-Tandem Mass Spectrometry Method for Detecting Adrenocortical Hormones and Establishment of Age-Stratified Reference Intervals in Reproductive-Aged Women from Guangxi, China
Yixuan LIU ; Tingwei JIN ; Yushuang WEI ; Xuelian QIN ; Siyu DENG ; Jie ZHENG ; Boteng YAN ; Yuanyuan NONG ; Yu YE ; Shengzhu HUANG ; Yu LONG ; Jianmin LI ; Ganqin WANG ; Pei HUANG ; Jinghang JIANG ; Fan WU ; Zengnan MO ; Yonghua JIANG
Annals of Laboratory Medicine 2026;46(2):146-154
Background:
Adrenocortical hormones, particularly 11-oxygenated androgens, are pivotal in female reproductive health and fertility. Standardized detection kits and population-specific reference intervals are lacking in China, hindering related clinical applications.
Methods:
A HPLC-tandem mass spectrometry (HPLC-MS/MS) pipeline was developed, rigorously validated, and applied to simultaneously quantify corticosterone, cortisone, cortisol, 18-OH cortisol, androstenedione (A4), 11β-hydroxyandrostenedione (11-OH A4), dehydroepiandrosterone, and dehydroepiandrosterone sulfate in serum samples from 455 reproductive-aged women (18–45 yrs) in Guangxi, China. Age-dependent concentration trends were analyzed, and reference intervals stratified by age (2.5th to 97.5th percentiles) were established. Correlations with body-composition metrics, ethnicity, and the menstrual cycle were investigated.
Results:
The HPLC-MS/MS method demonstrated high precision (intra- and inter-assay CVs < 15%), accuracy, and sensitivity. All eight hormones exhibited significant age-related declines (P < 0.001 for seven hormones; P = 0.001 for 11-OH A4). Notably, 11-OH A4 levels were significantly lower in the 35–45-yr (3.05 nmol/L) and 25–34-yr (3.09 nmol/L) age groups than in the 18–24-yr (3.57 nmol/L) age group, whereas no significant difference was observed between the 35–45-yr and 25–34-yr age groups. Weak negative correlations were observed between the body mass index and corticosterone and cortisone levels, whereas ethnicity and the menstrual cycle showed no significant associations with hormone levels.
Conclusions
We developed an HPLC-MS/MS-based method for simultaneously quantifying eight adrenocortical hormones, including 11-OH A4, and defined age-specific reference intervals for reproductive-aged Chinese women. These findings advance the clinical utility of adrenocortical hormones in diagnosing and managing reproductive disorders.
4.Chinese expert consensus on the diagnosis and treatment of chronic pain after lung surgery with integrated Traditional Chinese and Western medicine (2026 edition)
Jichen QU ; Wentian ZHANG ; Jianqiao CAI ; Zhigang CHEN ; Bin LI ; Wei DAI ; Xiangwu WANG ; Yan LI ; Xiang LÜ ; ; Yongfu ZHU ; Mingran XIE ; Sufang ZHANG ; Lei JIANG
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2026;33(04):522-534
Chronic post-surgical pain (CPSP) is a common long-term complication following lung surgery. Its high incidence significantly impacts patients’ quality of life and functional recovery, and imposes a substantial socioeconomic burden. This consensus aims to systematically establish a standardized integrated Chinese and Western medicine diagnostic and treatment framework for chronic post-lung surgery pain (CPLSP). Based on the latest domestic and international evidence-based medical research and multidisciplinary clinical experience, the working group comprehensively elaborates on core issues regarding CPLSP, including its definition, epidemiology, pathogenesis, clinical assessment, Western medical treatment, traditional Chinese medicine (TCM) treatment, and integrated strategies. The consensus emphasizes a patient-centered approach, adhering to the principles of multimodality, individualization, and stepwise management, highlighting the synergistic advantages of integrating Chinese and Western medicine throughout the entire perioperative management cycle encompassing "perioperative anti-inflammation, acute analgesia, and chronic rehabilitation." Through systematic literature retrieval and evidence integration, a total of 9 core recommendations were established to provide scientifically sound and clinically practical guidance.
5.Mechanistic study of mitochondrial dysfunction in renal injury induced by maternal bone lead mobilization during pregnancy in rats
Ling LI ; Lin ZHANG ; Li LI ; Yuting WEI ; Man LYU ; Zeshi ZHANG ; Li MA ; Anxin LU ; Yin LIN ; Shaohua WANG ; Chonghuai YAN
Journal of Environmental and Occupational Medicine 2026;43(3):286-292
Background Lead is a typical persistent environmental pollutant that can accumulate in bones for decades. During pregnancy, alterations in calcium metabolism promote the mobilization of bone lead, resulting in secondary exposure; however, the mechanisms by which pregnancy-associated bone lead mobilization affects maternal renal function remain unclear. Objective To investigate the role of mitochondrial dysfunction in pregnancy-related bone lead mobilization-induced renal injury. Methods Newly weaned female Wistar rats were randomly assigned to a control or a lead-exposed group administered either 0.05% sodium acetate or 0.05% lead acetate in drinking water. Following a 4-week lead exposure and a 4-week washout period, the females were co-housed with healthy age-matched males for mating. Rats were sacrificed at early (gestational day 3) and late (gestational day 17) pregnancystages, respectively. Renal histopathology was assessed using hematoxylin and eosin staining staining. Mitochondria-related indicators, including oxidative stress, inflammatory responses, and energy metabolism, were measured. Differential metabolites were identified using serum metabolomics. Results Renal injury in the lead-exposed pregnant rats progressed in a time-dependent manner, characterized by degeneration of proximal tubular epithelial cells, glomerular hyaline changes, and interstitial inflammatory cell infiltration. Repeated measures ANOVA indicated a significant interaction between the treatment factor (lead exposure) and the temporal factor (gestational stage) on renal injury (P<0.001). Further analysis of mitochondrial function-related indicators in late-pregnancy renal tissue revealed that the lead exposure group exhibited significantly increased levels of malondialdehyde (MDA) and reactive oxygen species (ROS) (P<0.05), accompanied by a reduction in superoxide dismutase (SOD) and reduced glutathione (GSH) activities (P<0.05); regarding inflammatory markers, levels of interleukin-18 (IL-18) and interleukin-1β (IL-1β) were elevated (P<0.01), whereas interleukin-33 (IL-33) was decreased in the lead-exposed group (P<0.05); energy metabolism-related indicators, including adenosine triphosphate (ATP) level, Na+-K+-ATPase and Ca2+-Mg2+-ATPase activities, and mitochondrial respiratory chain complexes I, III, and V activities, were significantly reduced (P<0.05) in the lead-exposed gorup. The typical differential metabolite N-methylisoleucine, identified through serum metabolomics analysis, was negatively correlated with blood lead levels, kidney injury scores, and IL-1β, while positively correlated with catalase (CAT) activity and Ca2+-Mg2+-ATPase. Conclusions Mitochondrial dysfunction may play a critical role in renal injury induced by bone lead mobilization during late gestation.
6.Material Basis of Anti-Inflammatory Efficacy and Mechanism of Action of Bushen Tongdu Prescription Based on UPLC-LTQ-Orbitrap-MS and Network Pharmacology
Yan RONG ; Lulu JING ; Hongping HOU ; Huijun WANG ; Lihua CHEN ; Yunxin CHEN ; Liang LI ; Li LIN ; Xiaoqin LUO ; Haiyu ZHAO ; Xiaolu WEI
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(10):152-161
ObjectiveThis paper aims to investigate the material basis of the anti-inflammatory efficacy and mechanism of action of Bushen Tongdu prescription (BSTDP). MethodsThe chemical components of BSTDP and its blood-absorbed components in vivo were systematically identified by using ultra-performance liquid chromatography-linear ion trap-electrostatic field orbitrap high-resolution mass spectrometry (UPLC-LIT-Orbitrap-MS). Network pharmacology was employed to screen blood-absorbed bioactive components and potential targets of this formula. A protein-protein interaction (PPI) network of core targets was constructed to conduct enrichment analysis. Molecular docking was further utilized to verify the binding affinity between key components and targets. The inflammatory model was established and verified in vivo by using a transgenic zebrafish Tg (mpx: GFP). At three days post-fertilization (3 dpf), larvae of zebrafish were randomly assigned to blank group, model group, positive drug dexamethasone acetate group (75 μmol·L-1), and BSTDP groups with low, medium, and high doses (500, 1 000, and 2 000 mg·L-1). The distribution and quantity of neutrophils in the yolk sac region were observed under a fluorescence microscope. The mRNA expression levels of key genes in the toll-like receptor 4 (TLR4)/myeloid differentiation factor 88 (MyD88)/nuclear factor kappa-B (NF-κB) signaling pathway and inflammatory factors including interleukin (IL)-1β, IL-6, and tumor necrosis factor-α (TNF-α) were detected by Real-time quantitative polymerase chain reaction (Real-time PCR). ResultsA total of 120 chemical components were identified in BSTDP, among which 26 original components were confirmed by using serum pharmacochemical methods. A total of 227 common targets linking rheumatoid arthritis (RA) and the blood-absorbed components were screened by network pharmacology. It is suggested that pseudobrucine, vomicine, sinapine, rehmannioside, cinnamyl alcohol glycoside, and methylephedrine exert anti-inflammatory effects by acting on core targets including protein kinase B1 (Akt1), signal transducer and activator of transcription 3 (STAT3), tumor necrosis factor (TNF), TLR4, mitogen-activated protein kinase 14 (MAPK14), and phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit α (PIK3CA), thereby modulating multiple signaling pathways such as TLR4 and NF-κB. In vivo verification in zebrafish demonstrates that the maximum tolerable concentration of Bushen Tongdu Formula is 2 000 mg·L-1. Compared to those in the blank group, zebrafish in the model group showed a significantly higher number of neutrophils in the yolk sac region (P<0.01) and rising mRNA levels of TLR4, MyD88, NF-κB, TNF-α, IL-6, and IL-1β (P<0.01). Compared to that in the model group, the number of neutrophils was significantly reduced in BSTDP groups with medium and high doses, as well as the dexamethasone acetate group (P<0.05, P<0.01). There was no statistically significant difference in the low dose group. The mRNA expression levels of TLR4, MyD88, NF-κB, TNF-α, IL-6, and IL-1β were significantly down-regulated (P<0.05, P<0.01). ConclusionThis paper identifies the material basis of the efficacy of BSTDP, demonstrating that the formula can exert an anti-inflammatory effect through the TLR4/MyD88/NF-κB signaling pathway. The results provide scientific experimental evidence for its further clinical application.
7.Material Basis of Anti-Inflammatory Efficacy and Mechanism of Action of Bushen Tongdu Prescription Based on UPLC-LTQ-Orbitrap-MS and Network Pharmacology
Yan RONG ; Lulu JING ; Hongping HOU ; Huijun WANG ; Lihua CHEN ; Yunxin CHEN ; Liang LI ; Li LIN ; Xiaoqin LUO ; Haiyu ZHAO ; Xiaolu WEI
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(10):152-161
ObjectiveThis paper aims to investigate the material basis of the anti-inflammatory efficacy and mechanism of action of Bushen Tongdu prescription (BSTDP). MethodsThe chemical components of BSTDP and its blood-absorbed components in vivo were systematically identified by using ultra-performance liquid chromatography-linear ion trap-electrostatic field orbitrap high-resolution mass spectrometry (UPLC-LIT-Orbitrap-MS). Network pharmacology was employed to screen blood-absorbed bioactive components and potential targets of this formula. A protein-protein interaction (PPI) network of core targets was constructed to conduct enrichment analysis. Molecular docking was further utilized to verify the binding affinity between key components and targets. The inflammatory model was established and verified in vivo by using a transgenic zebrafish Tg (mpx: GFP). At three days post-fertilization (3 dpf), larvae of zebrafish were randomly assigned to blank group, model group, positive drug dexamethasone acetate group (75 μmol·L-1), and BSTDP groups with low, medium, and high doses (500, 1 000, and 2 000 mg·L-1). The distribution and quantity of neutrophils in the yolk sac region were observed under a fluorescence microscope. The mRNA expression levels of key genes in the toll-like receptor 4 (TLR4)/myeloid differentiation factor 88 (MyD88)/nuclear factor kappa-B (NF-κB) signaling pathway and inflammatory factors including interleukin (IL)-1β, IL-6, and tumor necrosis factor-α (TNF-α) were detected by Real-time quantitative polymerase chain reaction (Real-time PCR). ResultsA total of 120 chemical components were identified in BSTDP, among which 26 original components were confirmed by using serum pharmacochemical methods. A total of 227 common targets linking rheumatoid arthritis (RA) and the blood-absorbed components were screened by network pharmacology. It is suggested that pseudobrucine, vomicine, sinapine, rehmannioside, cinnamyl alcohol glycoside, and methylephedrine exert anti-inflammatory effects by acting on core targets including protein kinase B1 (Akt1), signal transducer and activator of transcription 3 (STAT3), tumor necrosis factor (TNF), TLR4, mitogen-activated protein kinase 14 (MAPK14), and phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit α (PIK3CA), thereby modulating multiple signaling pathways such as TLR4 and NF-κB. In vivo verification in zebrafish demonstrates that the maximum tolerable concentration of Bushen Tongdu Formula is 2 000 mg·L-1. Compared to those in the blank group, zebrafish in the model group showed a significantly higher number of neutrophils in the yolk sac region (P<0.01) and rising mRNA levels of TLR4, MyD88, NF-κB, TNF-α, IL-6, and IL-1β (P<0.01). Compared to that in the model group, the number of neutrophils was significantly reduced in BSTDP groups with medium and high doses, as well as the dexamethasone acetate group (P<0.05, P<0.01). There was no statistically significant difference in the low dose group. The mRNA expression levels of TLR4, MyD88, NF-κB, TNF-α, IL-6, and IL-1β were significantly down-regulated (P<0.05, P<0.01). ConclusionThis paper identifies the material basis of the efficacy of BSTDP, demonstrating that the formula can exert an anti-inflammatory effect through the TLR4/MyD88/NF-κB signaling pathway. The results provide scientific experimental evidence for its further clinical application.
8.Genetic analysis and prenatal diagnosis of structural brain abnormalities associated with TUBB gene c.155A>G variant.
Yifan LIU ; Wei SONG ; Xinlian WANG ; Yan RUAN ; Meng ZHANG ; Yujiao CHEN ; Yan LIU ; Puqing ZHANG ; Li WANG ; Yousheng YAN
Chinese Journal of Medical Genetics 2026;43(2):136-142
OBJECTIVE:
To explore the genotype-phenotype correlation in a Chinese family with structural brain abnormalities due to variant of the TUBB gene.
METHODS:
A family undergoing prenatal diagnosis at Beijing Obstetrics and Gynecology Hospital in October 2024 was selected as the study subject. Clinical data were collected. Amniotic fluid sample was subjected to chromosomal copy number variation sequencing (CNV-seq). Trio whole-exome sequencing (Trio-WES) was carried out on the amniotic fluid and parental blood samples, and candidate variant was verified by Sanger sequencing. This study was approved by the Medical Ethics Committee of the hospital (Ethics No.: 2023-KY-076-01).
RESULTS:
Both prenatal ultrasound and fetal MRI showed deviation of brain midline, unilateral lateral ventriculomegaly, and bilateral gyral asymmetry. Trio-WES revealed that the fetus has harbored a maternally derived heterozygous missense variant of the TUBB gene [NM_178014.4: c.155A>G (p.N52S)]. Sanger sequencing confirmed that the woman and a previously terminated fetus both harbored the same variant. Both the proband and two fetuses exhibited similar neuroimaging abnormalities including midline deviation and asymmetrical gyri. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the variant was classified as likely pathogenic (PM2_Supporting+PS2_Moderate+PS3).
CONCLUSION
The heterozygous c.155A>G (p.N52S) variant was the TUBB gene probably underlay the pathogenesis of the structural brain abnormalities in this family. Above findings have expanded the phenotypic spectrum associated with the variant and facilitated the prenatal diagnosis for this family.
Humans
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Female
;
Pregnancy
;
Prenatal Diagnosis
;
Tubulin/genetics*
;
Adult
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Brain/diagnostic imaging*
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Male
;
Pedigree
;
DNA Copy Number Variations/genetics*
;
Exome Sequencing
;
Genetic Association Studies
;
Magnetic Resonance Imaging
9.Expert Consensus on Clinical Application of Qidong Yixin Oral Liquid
Changkuan FU ; Xiaochang MA ; Mingjun ZHU ; Yue DENG ; Hongxu LIU ; Mingxue ZHANG ; Ying CHEN ; Yan ZHOU ; Ling ZHANG ; Jianhua FU ; Wei YANG ; Yu'er HU ; Ming CHEN ; Yanming XIE ; Yuanyuan LI
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(4):147-158
The prescription of Qidong Yixin oral liquid is derived from the experience of national medical master Ren Jixue in treating viral myocarditis (VMC). It has the functions of tonifying Qi, nourishing the heart,calming the mind, and relieving palpitations. It is used to treat VMC and angina pectoris of coronary heart disease caused by deficiency of both Qi and Yin. However,the understanding of its efficacy evidence, advantageous aspects, dosage and administration, and medication safety remains insufficient in clinical practice. Therefore,the development of the Expert Consensus on the Clinical Application of Qidong Yixin Oral Liquid (hereinafter referred to as consensus) was initiated. Consensus strictly followed the process and methods of the expert consensus on the clinical application of Chinese patent medicines of the China Association of Chinese Medicine,successively completing multiple tasks such as the consensus project initiation,determination of clinical problems,evidence search and evaluation,formation of recommendation opinions and consensus suggestions,solicitation of opinions,peer review, submission for review and release, and so on. Consensus formed a total of 10 recommendation opinions and 12 consensus suggestions,clarifying the clinical positioning,efficacy advantages,syndrome differentiation,dosage and administration,combination therapy,timing of medication,adverse reactions,contraindications, and precautions of Qidong Yixin oral liquid,indicating that it has good clinical advantages and safety in the treatment of VMC and angina pectoris of coronary heart disease,providing norms and references for physicians to safely and rationally apply Qidong Yixin oral liquid. Consensus was reviewed and approved for release by the Standardization Office of the China Association of Chinese Medicine on December 23, 2024. Standard number:GSCACM-376-2024.
10.Sanren Runchang Formula Regulates Brain-gut Axis to Treat IBS-C: A Randomized Controlled Trial
Teng LI ; Xinrong FAN ; He YAN ; Zhuozhi GONG ; Mengxi YAO ; Na YANG ; Yuhan WANG ; Huikai HU ; Wei WEI ; Tao LIU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(2):154-161
ObjectiveTo observe the clinical efficacy of Sanren Runchang formula in treating constipation-predominant irritable bowel syndrome (IBS-C) by regulating the brain-gut axis and the effects of the formula on serum levels of 5-hydroxytryptamine (5-HT), vasoactive intestinal peptide (VIP), and substance P (SP). MethodsA randomized controlled design was adopted, and 72 IBS-C patients meeting Rome Ⅳ criteria were randomized into observation and control groups (36 cases).The observation group received Sanren Runchang formula granules twice daily, and the control group received lactulose oral solution daily for 4 weeks. IBS Symptom Severity Scale (IBS-SSS), IBS Quality of Life Scale (IBS-QOL), and Bristol Stool Form Scale (BSFS) were used to assess clinical symptoms, and bowel movement frequency was recorded. The Self-Rating Anxiety Scale (SAS) and Self-Rating Depression Scale (SDS) were employed to evaluate psychological status. ELISA was employed to measure the serum levels of 5-HT, VIP, and SP. ResultsThe total response rate in the observation group was 91.67% (33/36), which was higher than that (77.78%, 28/36) in the control group (χ2=4.50, P<0.05). After treatment, both groups showed increased defecation frequency and BSFS scores, decreased IBS-SSS total score, abdominal pain and bloating scores, IBS-QOL health anxiety, anxiety, food avoidance, and behavioral disorders scores, SAS and SDS scores, serum 5-HT and VIP levels, and increased SP levels (P<0.05, P<0.01). Moreover, the observation group showed more significant changes in the indicators above than the control group (P<0.05, P<0.01). The SP level showed no significant difference between the two groups. During the 4-week follow-up, the recurrence rate was 5.88% in the observation group and 31.25% in the control group. No adverse events occurred in observation group, and 2 cases of mild diarrhea occurred in the control group. ConclusionSanren Runchang formula demonstrated definitive efficacy in alleviating gastrointestinal symptoms and improving the psychological status and quality of life in IBS-C patients, with a low recurrence rate. The formula can regulate serum levels of neurotransmitters such as 5-HT and VIP, suggesting its potential regulatory effect on the brain-gut axis through modulating neurotransmitters and neuropeptides. However, its complete mechanism of action requires further investigation through detection of additional brain-gut axis-related biomarkers.

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