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.Progress in study of effect of valproic acid on male reproductive function in epileptic adults
Kejing LI ; Pei LI ; Zheng YIN ; Xijun LI ; Xin LI
Chinese Journal of Reproduction and Contraception 2025;45(8):851-855
Valproic acid (VPA) is a traditional broad-spectrum antiepileptic drug, but long-term use of VPA will increase the risk of adverse drug reactions, with metabolic syndrome being the most common and often leading to infertility in men. Therefore, it is crucial to explore the relationship between epilepsy, VPA, metabolic syndrome and reproduction, the underlying pathogenesis, risk factors, and therapeutic measures. This exploration is essential for preventing adverse drug reactions and for guiding safe and rational use of drugs in clinical practice. This article summarizes the incidence of drug-induced metabolic syndrome and infertility in men with epilepsy who are treated with oral VPA, aiming to provide new research insights for optimizing treatment and reducing drug side effects.
5.Danggui Yinzi prescription combined with focused ultrasound in treatment of vulvar lichen sclerosus
Yingping LIU ; Jieqiong CAO ; Caifeng XU ; Chongchong PEI ; Yan WANG ; Jun ZHENG
China Modern Doctor 2025;63(11):42-44
Objective To observe clinical efficacy of Danggui Yinzi prescription combined with focused ultrasound in treatment of vulvar lichen sclerosus(VLS)with blood deficiency and dryness.Methods A total of 60 patients with blood deficiency and dryness VLS hospitalized in Gansu Provincial Hospital of Traditional Chinese Medicine from September 2023 to September 2024 were selected as study subjects,they were divided into control group(n=30)and treatment group(n=30)by random number table method.Control group was treated with focused ultrasound,and treatment group was treated with oral Danggui Yinzi prescription on the basis of control group.After 1 month of treatment,symptom scores(skin lesion area,color,etc.)and clinical efficacy of two groups were compared.Results After treatment,symptom scores of two groups were lower than those before treatment,and symptom scores of treatment group were lower than those of control group.Total effective rate of treatment group was higher than that of control group(P<0.05).Conclusion Danggui Yinzi prescription combined with focused ultrasound is effective in treatment of VLS with blood deficiency and dryness,which can significantly reduce or eliminate itching symptoms and promote skin healing.
6.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.
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.Autophagy in erectile dysfunction: focusing on apoptosis and fibrosis.
Pei-Yue LUO ; Jun-Rong ZOU ; Tao CHEN ; Jun ZOU ; Wei LI ; Qi CHEN ; Le CHENG ; Li-Ying ZHENG ; Biao QIAN
Asian Journal of Andrology 2025;27(2):166-176
In most types of erectile dysfunction, particularly in advanced stages, typical pathological features observed are reduced parenchymal cells coupled with increased tissue fibrosis. However, the current treatment methods have shown limited success in reversing these pathologic changes. Recent research has revealed that changes in autophagy levels, along with alterations in apoptosis and fibrosis-related proteins, are linked to the progression of erectile dysfunction, suggesting a significant association. Autophagy, known to significantly affect cell fate and tissue fibrosis, is currently being explored as a potential treatment modality for erectile dysfunction. However, these present studies are still in their nascent stage, and there are limited experimental data available. This review analyzes erectile dysfunction from a pathological perspective. It provides an in-depth overview of how autophagy is involved in the apoptotic processes of smooth muscle and endothelial cells and its role in the fibrotic processes occurring in the cavernosum. This study aimed to develop a theoretical framework for the potential effectiveness of autophagy in preventing and treating erectile dysfunction, thus encouraging further investigation among researchers in this area.
Male
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Humans
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Autophagy/physiology*
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Apoptosis/physiology*
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Erectile Dysfunction/physiopathology*
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Fibrosis
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Penis/pathology*
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Animals
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Endothelial Cells/pathology*
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Myocytes, Smooth Muscle/pathology*
9.Monitoring and model prediction of human brucellosis in Tangshan City, Hebei Province from 2016 to 2024
Xiangbo LIU ; Wen GAO ; Renjie E ; Ling ZHANG ; Guangyue XIE ; Jie PEI ; Hui WANG ; Zheng LIU ; Hongli LIU
Chinese Journal of Infectious Diseases 2025;43(4):232-238
Objective:To analyze the epidemic characteristics of human brucellosis in Tangshan City, Hebei Province from 2016 to 2024, and to establish a prediction model for forecasting incidence of human brucellosis in Tangshan City from 2025 to 2028, so that to provide evidence for prevention and control strategies.Methods:The incidence data of human brucellosis in Tangshan City from 2016 to 2024 were collected. Brucella strains isolated from blood cultures of patients with acute brucellosis were identified.The onset time and demographic distributions of brucellosis were analyzed using descriptive epidemiological methods. Chi-square test was used for statistical analysis. Python software was used to establish a seasonal autoregressive integrated moving average model (SARIMA model) and predict the incidence of brucellosis in Tangshan City from 2025 to 2028. Results:From 2016 to 2024, a total of 2 446 cases of human brucellosis in Tangshan City were reported, with the highest incidence in 2016 (378 cases) and the lowest in 2022 (277 cases).Seasonal variation was observed, with 54.87%(1 342/2 446) occurring in spring and summer (March to July). The incidence rate of male was 5.28/100 000, which was significantly higher than that of female (1.94/100 000) ( χ2=554.96, P<0.001). The cases spanned all age groups, with the highest incidence among those aged 50 to 59 (30.25%(740/2 446)). Farmers engaged in cattle/sheep breeding accounting for 85.73% (2 097/2 446) of cases. A total of 236 blood samples were collected from patients with acute brucellosis, and 12 Brucella strains were isolated and identified as sheep type Ⅲ Brucella. The optimal model constructed was SARIMA (1, 0, 0) (1, 0, 1) 12, which was used to predict the incidence of human brucellosis in Tangshan City from 2025 to 2028. The results showed that the overall incidence was relatively stable, retaining the characteristic single annual peak. Conclusions:Human brucellosis in Tangshan City peaks in spring/summer and predominantly affects cattle/sheep farmers. The SARIMA (1, 0, 0) (1, 0, 1) 12 model effectively fits the epidemiological data of human brucellosis in Tangshan City well and enables reliable future trend predictions, supporting scientific and effective prevention and control work.
10.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.

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