1.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.
2.Luoshi Neiyi Prescription Treats Endometriosis Through TLR4/NF-κB Signaling Pathway
Yuanyuan RUAN ; Sai XU ; Jiangyue TANG ; Xiang LI ; You ZOU ; Fangli PEI ; Lizheng WU ; Kaidi ZHENG ; Shuhong LIN ; Weilan ZHONG ; Cheng ZENG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(19):185-196
ObjectiveTo investigate the mechanism by which Luoshi Neiyi prescription treats endometriosis (EMs) through the Toll-like receptor 4 (TLR4)/nuclear factor-κB (NF-κB) signaling pathway. MethodsAnimal experiments were conducted with 50 female SD rats, which were randomized into a sham operation group (10 rats) and a modeling group (40 rats). An autologous endometrial transplantation method was used for the modeling of EMs. The 36 successfully modeled rats were randomly allocated into four groups (n=9 each): The model group, the low-dose (7.87 g·kg-1) Luoshi Neiyi prescription group, the high-dose (15.74 g·kg-1) Luoshi Neiyi prescription group, and the dienogest (0.20 mg·kg-1) group. The physiological status and ectopic lesion volumes of rats in each group were observed. Hematoxylin-eosin (HE) staining was used to observe the morphology of the eutopic endometrial tissue. Enzyme-linked immunosorbent assay (ELISA) was employed to measure the levels of inflammatory factors including interleukin-1β (IL-1β) and prostaglandin E2 (PGE2) in the serum of EMs rats. Immunohistochemistry was used to detect the expression of matrix metalloproteinase-9 (MMP-9) and vascular endothelial growth factor A (VEGFA) in the eutopic endometrial tissue. Western blot was adopted to determine the protein levels of TLR4, myeloid differentiation factor 88 (MyD88), phosphorylated nuclear factor-κB (p-NF-κB)/NF-κB, MMP-9, and VEGFA in the eutopic endometrial tissue. In the cell experiments, the cell-counting kit-8 (CCK-8) assay was employed to screen the optimal concentration of Luoshi Neiyi prescription-containing serum, and a scratch assay was performed to assess the migration ability of iheESCs cells. Interventions with Luoshi Neiyi prescription-containing serum, resatorvid (TAK-242, a TLR4 inhibitor), and lipopolysaccharides (LPS, a TLR4 agonist) were conducted, and Western blot was used to detect the expression of proteins related to the TLR4/NF-κB signaling pathway. ResultsThis experiment successfully replicated 36 EMs rat models. Compared with the sham operation group, the model group exhibited visible ectopic lesions on the abdominal wall and an increase in the writhing response score (P<0.01). Furthermore, HE staining revealed the model group exhibited a thickened eutopic epithelium, stromal cell disarrangement, and evident infiltration of inflammatory cells. In addition, the model group showed elevated serum levels of IL-1β and PGE2 (P<0.05, P<0.01), increased positive expression of MMP-9 and VEGFA in the ectopic endometrial tissue (P<0.01), and up-regulated protein levels of TLR4, MyD88, p-NF-κB/NF-κB, MMP-9, and VEGFA in the ectopic endometrial tissue (P<0.05, P<0.01). Compared with the model group, all treatment groups exhibited a reduction in the ectopic lesion volume (P<0.01). Furthermore, the writhing response scores were decreased in the high-dose Luoshi Neiyi prescription group and the dienogest group (P<0.05, P<0.01). The pathological state of the ectopic endometrial tissue was alleviated to varying degrees in the treatment groups. Low-dose Luoshi Neiyi prescription reduced serum PGE2 levels, and high-dose Luoshi Neiyi prescription and dienogest decreased serum IL-1β and PGE2 levels in EMs rats (P<0.05, P<0.01). The treatment groups showed decreased positive expression of MMP-9 and VEGFA in the ectopic endometrial tissue (P<0.05, P<0.01) and down-regulated protein levels of TLR4, MyD88, p-NF-κB/NF-κB, MMP-9, and VEGFA in the ectopic endometrial tissue (P<0.05, P<0.01). The cell experiments showed that 5%, 10%, and 20% Luoshi Neiyi prescription-containing sera significantly reduced the viability and inhibited the migration of iheESCs. Compared with the control group, 5%, 10%, and 20% Luoshi Neiyi prescription-containing serum groups and the TAK-242 group showed reduced protein levels of TLR4, MyD88, p-NF-κB/NF-κB, MMP-9, and VEGFA (P<0.01). Compared with the control group, the LPS group showed increased expression of the above proteins (P<0.01). Compared with the LPS group, the LPS+5%, 10%, and 20% Luoshi Neiyi prescription-containing serum groups showed reduced expression of the above proteins (P<0.05, P<0.01). ConclusionLuoshi Neiyi prescription may modulate the TLR4/NF-κB signaling pathway to reduce the inflammatory response and histopathological damage in the eutopic endometrium and suppress the adhesive, invasive, and angiogenic biological processes in the ectopic endometrial tissue, thereby exerting its therapeutic effect on EMs.
3.Screening of the Monkeypox Virus Key Proteins Regulating Inflammation Based on NOD-like Receptor Family Pyrin Domain Containing 3 Inflammasome Signaling
Mingxing LUO ; Chenchen PEI ; Liyun GONG ; Xinlei ZHENG ; Jun QIAN ; Quan LIU ; Jun ZHENG
Journal of Sun Yat-sen University(Medical Sciences) 2026;47(2):259-268
ObjectiveTo screen and identify the key proteins of Monkeypox Virus (MPXV) that regulate the NOD-like receptor family pyrin domain containing 3 (NLRP3) inflammasome. MethodsUsing the genomic sequence of the domestic epidemic strain of MPXV as a template to construct an expression plasmid library containing 82 viral genes. To screen for viral genes that significantly inhibit the production of interleukin-1β (IL-1β), the expression plasmids encoding NLRP3, apoptosis-associated speck-like protein containing a CARD (ASC), cysteine protease-1(Caspase-1), and pro-IL-1β-Luc reporter gene were co-transfected into HEK293T cells, along with the viral gene plasmids. The signal of the NLRP3 inflammasome was monitored by measuring luciferase activity in the cell supernatant. The inhibition effects were further confirmed by Western Blot (WB), and the interactions between the viral proteins and the components of the NLRP3 inflammasome were determined by Co-immunoprecipitation (Co-IP). Additionally, site-directed mutagenesis was performed to identify the specific interaction motifs. ResultsAn NLRP3 inflammasome signal screening system were successfully established and optimized. Screening the MPXV plasmids using this system revealed that B10R most significantly inhibits IL-1β maturation in a dose-dependent manner (P<0.000 1). Further characterization demonstrated that B10R interacts directly with pro-IL-1β. Notably, the C-terminal motif mutant of B10R (B10Rmut-C, T96A, Y97A, I98A) lost its ability to interact with pro-IL-1β. ConclusionsThe MPXV B10R protein interacts with pro-IL-1β, thereby inhibiting the IL-1β maturation. The C-terminal motif (T96, Y97, I98) of the B10R protein is crucial for function. This study provides important clues and a foundation for further elucidating the molecular mechanisms by which MPXV regulates the NLRP3 inflammasome.
4.Exploring mechanism of action of hypericin in antidepressant effects based on single-cell sequencing
Hui-xin NI ; Hai-xin LIU ; Bing-can ZHOU ; Ming-heng CHEN ; Ping-yan LIN ; Zheng-tao GAO ; Xin-pei LIN ; Yao LIN ; Fang-zhen WU ; Qian XU
Chinese Pharmacological Bulletin 2025;41(5):837-843
Aim To investigate the antidepressant mechanism of hyperforin via the utilization of single-cell sequencing technology.Methods C57BL/6 mice were randomly divided into the control group,depres-sion model group,and hyperforin intervention group.The chronic unpredictable mild stress(CUMS)model was induced and drug interventions were administered for 28 d.Behavioral experiments were conducted to as-sess depressive symptoms,and hippocampal tissue was collected for single-cell RNA sequencing.Key cell populations and differentially expressed genes across groups were identified,followed by PPI network,GO,and KEGG enrichment analysis.Results Behavioral experiments indicated that CUMS successfully induced depressive symptoms in mice,while hyperforin im-proved depressive behavior.In the depression model group,the proportion of brain perivascular macrophages(PVM)increased,and this proportion decreased after hyperforin intervention,approaching the level seen in the control group.The top 20 common differentially ex-pressed genes in the PVM subpopulation were Saa3,Hbb-bs and Ccl24.PPI network analysis identified core targets,including Ccl2,Dhx9,C3,Msr1,Cxcl2 and Cx3cr1.KEGG enrichment analysis revealed pathways related to chemokines,phagosome formation,and inosi-tol phosphate metabolism.Conclusion The antide-pressant mechanism of hyperforin may be related to the regulation of Ccl24 and its related chemokine signaling pathway by PVM.
5.Research progress in radiation-induced esophageal injury
Qiang FU ; Yu LIN ; Fei ZHENG ; Yuanji XU ; Wenji XUE ; Ye ZHANG ; Qifeng WANG ; Jinbo YUE ; Pei YANG ; Wencheng ZHANG ; Junqiang CHEN
Chinese Journal of Radiation Oncology 2025;34(9):874-881
Radiation-induced esophageal injury (RIEI) is a frequent complication following radiotherapy for thoracic and head-neck malignancies, which may lead to severe sequelae including esophageal stricture and perforation, adversely affecting patients' quality of life and therapeutic outcomes. With advancements in radiotherapy techniques — particularly the adoption of unconventional fractionation regimens, concurrent chemoradiotherapy, and combined molecular targeted / immunotherapy — the incidence of RIEI has been increasing. In this review, recent advances in understanding the pathogenesis, clinical manifestations, risk factors, and management strategies for RIEI were comprehensively summarized. Current therapeutic approaches have evolved beyond conventional anti-inflammatory and nutritional support to include novel interventions such as targeted therapy, free radical scavengers, and microbiota modulation, etc. Future research should prioritize the development of optimized, individualized prevention and treatment protocols to mitigate RIEI risk and improve patient prognosis.
6.Impact of continuous light exposure on gut microbiota of mice
Yan LI ; Lu ZHENG ; Ru TANG ; Jian WEI ; Pei SHI ; Zhan LI ; Youqin WANG
Chinese Journal of Pathophysiology 2025;41(2):322-330
AIM:To observe the effect of continuous light on the structure and differential metabolites of gut microbiota in mice.METHODS:The mice were randomly divided into normal light(light/dark,LD)group and 24-hour continuous light(light/light,LL)group.The body weight,fasting blood glucose,serum free fatty acids,serum triacylglycerol and serum total cholesterol levels of each group of mice were measured after 10 weeks.Fresh feces were collected,and 16S rRNA sequencing technology was used to study the effect of continuous light on the diversity,structure,and species composition of gut microbiota in mice.Additionally,liquid chromatography-mass spectrometry(LC-MS)analysis was per-formed to observe the effect of continuous light on the metabolites in mice.RESULTS:Compared with the LD group,the body weight,fasting blood glucose and lipid levels of the LL group were increased(P<0.05).At the phylum level,the proportion of Firmicutes in the LL group increased,while the proportion of Bacteroidetes decreased.At the class level,the abundance of norank_f_Muribaculaceae and Prevotellaceae_UCG-001 in the LL group decreased significantly,while the abundance of Lactobacillus,Turicibacter and Odoribacter increased significantly.Non-targeted metabolomics analysis iden-tified 65 and 73 differential metabolites under positive and negative modes,involving six major metabolic pathways,in-cluding ABC transporters,purine metabolism,pyrimidine metabolism,secondary bile acid biosynthesis,protein digestion and absorption,and choline metabolism in cancer.CONCLUSION:The structure and metabolites of gut microbiota in mice exposed to continuous light are relatively specific,and inosine may be a key biomarker and potential therapeutic tar-get for biological clock disorders.
7.International clinical practice guideline on the use of traditional Chinese medicine for functional dyspepsia (2025).
Sheng-Sheng ZHANG ; Lu-Qing ZHAO ; Xiao-Hua HOU ; Zhao-Xiang BIAN ; Jian-Hua ZHENG ; Hai-He TIAN ; Guan-Hu YANG ; Won-Sook HONG ; Yu-Ying HE ; Li LIU ; Hong SHEN ; Yan-Ping LI ; Sheng XIE ; Jin SHU ; Bin-Fang ZENG ; Jun-Xiang LI ; Zhen LIU ; Zheng-Hua XIAO ; Jing-Dong XIAO ; Pei-Yong ZHENG ; Shao-Gang HUANG ; Sheng-Liang CHEN ; Gui-Jun FEI
Journal of Integrative Medicine 2025;23(5):502-518
Functional dyspepsia (FD), characterized by persistent or recurrent dyspeptic symptoms without identifiable organic, systemic or metabolic causes, is an increasingly recognized global health issue. The objective of this guideline is to equip clinicians and nursing professionals with evidence-based strategies for the management and treatment of adult patients with FD using traditional Chinese medicine (TCM). The Guideline Development Group consulted existing TCM consensus documents on FD and convened a panel of 35 clinicians to generate initial clinical queries. To address these queries, a systematic literature search was conducted across PubMed, EMBASE, the Cochrane Library, China National Knowledge Infrastructure (CNKI), VIP Database, China Biology Medicine (SinoMed) Database, Wanfang Database, Traditional Medicine Research Data Expanded (TMRDE), and the Traditional Chinese Medical Literature Analysis and Retrieval System (TCMLARS). The evidence from the literature was critically appraised using the Grading of Recommendations Assessment, Development, and Evaluation (GRADE) approach. The strength of the recommendations was ascertained through a consensus-building process involving TCM and allopathic medicine experts, methodologists, pharmacologists, nursing specialists, and health economists, leveraging their collective expertise and empirical knowledge. The guideline comprises a total of 43 evidence-informed recommendations that span a range of clinical aspects, including the pathogenesis according to TCM, diagnostic approaches, therapeutic interventions, efficacy assessments, and prognostic considerations. Please cite this article as: Zhang SS, Zhao LQ, Hou XH, Bian ZX, Zheng JH, Tian HH, Yang GH, Hong WS, He YY, Liu L, Shen H, Li YP, Xie S, Shu J, Zeng BF, Li JX, Liu Z, Xiao ZH, Xiao JD, Zheng PY, Huang SG, Chen SL, Fei GJ. International clinical practice guideline on the use of traditional Chinese medicine for functional dyspepsia (2025). J Integr Med. 2025; 23(5):502-518.
Dyspepsia/drug therapy*
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Humans
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Medicine, Chinese Traditional/methods*
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Practice Guidelines as Topic
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Drugs, Chinese Herbal/therapeutic use*
8.Ionizing Radiation Alters Circadian Gene Per1 Expression Profiles and Intracellular Distribution in HT22 and BV2 Cells.
Zhi Ang SHAO ; Yuan WANG ; Pei QU ; Zhou Hang ZHENG ; Yi Xuan LI ; Wei WANG ; Qing Feng WU ; Dan XU ; Ju Fang WANG ; Nan DING
Biomedical and Environmental Sciences 2025;38(11):1451-1457
9.Alterations in striatal functional connectivity in schizophrenia patients with predominant negative symptoms
Yao ZHNAG ; Qin-yu LYU ; Xin-xin HUANG ; Chong-ze WANG ; Qi YAN ; Pei-juan WANG ; Zheng-hui YI
Fudan University Journal of Medical Sciences 2025;52(4):492-499
(rsFC)and their relationship with negative symptoms in schizophrenia patients with predominant negative symptoms(PNS).Methods Fifty-four schizophrenia patients with PNS and sixty-one healthy controls underwent resting-state functional magnetic resonance imaging(fMRI)scans.Data were collected on general demographic information,the Positive and Negative Syndrome Scale(PANSS),the Scale for the Assessment of Negative Symptoms(SANS),and the Temporal Experience of Pleasure Scale(TEPS).Twelve striatal subregions were selected as regions of interest(ROIs)to analyze the rsFC between each ROI and whole-brain voxels.The rsFC values of areas with significant differences were extracted for Pearson correlation analysis with negative symptoms.Results Compared with healthy controls,schizophrenia patients with PNS exhibited decreased rsFC between the right dorsal caudal putamen(DCP)and right insula,left middle frontal gyrus(MFG),right median cingulate and paracingulate gyri(MCC);between the left DCP and right putamen,left insula,left MFG;between the right dorsal rostral putamen(DRP)and bilateral MFG,left insula,right MCC;between the left DRP and right insula,left rolandic operculum;between the right ventral rostral putamen(VRP)and bilateral putamen,left MFG,right MCC;between the left VRP and right insula,left putamen,bilateral MFG,right MCC,left inferior parietal gyrus,excluding supramarginal and angular gyri.Decreased rsFC was also observed between the left ventral caudate/nucleus accumbens(inferior)and right insula,left anterior cingulate cortex,supracallosal,bilateral precuneus(a threshold of P<0.001 in voxel-level with P<0.05 in cluster-lever,corrected for family-wise error,PFWE<0.05/12=0.004).No regions showed increased rsFC in schizophrenia patients with PNS relative to healthy controls.And no significant correlations were found between striatal rsFC and negative symptoms(PBonferroni>0.05).Conclusion Schizophrenia patients with PNS exhibited widespread cortical-striatal functional connectivity abnormalities,particularly reduced rsFC between the putamen and the MFG,MCC and insula.
10.Exploring mechanism of action of hypericin in antidepressant effects based on single-cell sequencing
Hui-xin NI ; Hai-xin LIU ; Bing-can ZHOU ; Ming-heng CHEN ; Ping-yan LIN ; Zheng-tao GAO ; Xin-pei LIN ; Yao LIN ; Fang-zhen WU ; Qian XU
Chinese Pharmacological Bulletin 2025;41(5):837-843
Aim To investigate the antidepressant mechanism of hyperforin via the utilization of single-cell sequencing technology.Methods C57BL/6 mice were randomly divided into the control group,depres-sion model group,and hyperforin intervention group.The chronic unpredictable mild stress(CUMS)model was induced and drug interventions were administered for 28 d.Behavioral experiments were conducted to as-sess depressive symptoms,and hippocampal tissue was collected for single-cell RNA sequencing.Key cell populations and differentially expressed genes across groups were identified,followed by PPI network,GO,and KEGG enrichment analysis.Results Behavioral experiments indicated that CUMS successfully induced depressive symptoms in mice,while hyperforin im-proved depressive behavior.In the depression model group,the proportion of brain perivascular macrophages(PVM)increased,and this proportion decreased after hyperforin intervention,approaching the level seen in the control group.The top 20 common differentially ex-pressed genes in the PVM subpopulation were Saa3,Hbb-bs and Ccl24.PPI network analysis identified core targets,including Ccl2,Dhx9,C3,Msr1,Cxcl2 and Cx3cr1.KEGG enrichment analysis revealed pathways related to chemokines,phagosome formation,and inosi-tol phosphate metabolism.Conclusion The antide-pressant mechanism of hyperforin may be related to the regulation of Ccl24 and its related chemokine signaling pathway by PVM.

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