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.Liuwei Dihuangwan Reduces Valproic Acid-induced ASD-like Behaviors in Rats by Modulating mTOR-mediated Synaptic Plasticity in Prefrontal Cortex-hippocampus Circuit
Ji WU ; Niya YIN ; Lei OUYANG ; Siyu HE ; Fangyu PEI ; Ying XIE ; Fan LI ; Qian LEI ; Sha ZHAO ; Di ZHANG ; Qinquan ZHU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(22):1-11
ObjectiveTo investigate the effects of Liuwei Dihuangwan on mammalian target of rapamycin (mTOR)-mediated synaptic plasticity in the prefrontal cortex-hippocampus circuit in the rat model of valproic acid (VPA)-induced autism spectrum disorder (ASD) and to explore its potential mechanism for reducing behavioral abnormalities in ASD rats. MethodsThirteen SD pregnant rats were randomized into the VPA group (10 rats) and the control group (3 rats). On gestational day 13, the VPA group was intraperitoneally injected with VPA solution (600 mg·kg-1), while the control group was injected with an equal volume of normal saline. Male pups of 21 days old from both groups underwent behavioral tests to screen the ASD model rats (32 rats). These rats were then randomly assigned to the model (distilled water), vitamin D (1 480 U·kg-1), low-dose (0.75 g·kg-1) Liuwei Dihuangwan, and high-dose (1.5 g·kg-1) Liuwei Dihuangwan groups, and normal male pups were selected as the blank group (distilled water), with 8 rats in each group. Each group was treated with the corresponding drug or an equal volume of distilled water via gavage daily. After drug administration, behavioral tests were conducted again, followed by anesthesia and tissue collection. Nissl staining was carried out to observe the morphological changes of neurons in the prefrontal cortex and hippocampus. Transmission electron microscopy was used to observe changes in hippocampal synaptic structure. Immunohistochemistry and Western blot were employed to determine the expression of mTOR, phosphatidylinositol 3-kinase (PI3K), and protein kinase B (Akt) in the prefrontal cortex, as well as synaptophysin-1 (SYP-1), postsynaptic density 95 (PSD95), and N-methyl-D-aspartate receptors (NMDARs) in the hippocampus. ResultsCompared with the control group, the model group exhibited severe growth retardation and developmental deformities, with abnormal behavioral performance and pathological damage to neurons and synapses in the prefrontal cortex and hippocampus. Compared with the model group, all treatment groups showed reduced behavioral abnormalities and pathological damage to neurons and synapses in the prefrontal cortex and hippocampus, with the most significant effects in the high-dose Liuwei Dihuangwan group. Immunohistochemistry and Western blotting results showed that compared with the control group, the model group exhibited significantly increased protein levels and phosphorylation ratios of p-mTOR, p-Akt, and p-PI3K in the prefrontal cortex (P<0.05, P<0.01), and decreased protein levels of SYP-1 and PSD95 and increased protein level of NMDARs in the hippocampus (P<0.01). Compared with those in the model group, the protein levels of p-mTOR, p-Akt, and p-PI3K in the prefrontal cortex decreased in the vitamin D and high-dose Liuwei Dihuangwan groups (P<0.05), and the p-mTOR/mTOR, p-Akt/Akt, and p-PI3K/PI3K ratios in the prefrontal cortex declined in the vitamin D group (P<0.05). The high-dose Liuwei Dihuangwan group showed a reduction in p-PI3K/PI3K ratio (P<0.05). The protein level of SYP-1 and PSD95 in the hippocampus increased in the vitamin D, low-dose Liuwei Dihuangwan, and high-dose Liuwei Dihuangwan groups (P<0.05, P<0.01), while the protein level of NMDARs decreased in the vitamin D and high-dose Liuwei Dihuangwan groups (P<0.01). ConclusionLiuwei Dihuangwan significantly reduces VPA-induced ASD-like behaviors and enhances the regeneration and repair of neurons in the prefrontal cortex and synaptic structures in the hippocampus by downregulating of PI3K/Akt/mTOR expression in the prefrontal cortex, upregulating SYP-1 and PSD95 expression in the hippocampus, and inhibiting NMDARs expression.
3.Predictive value of MR myocardial strain in predicting recent adverse cardiovascular events after primary PCI in patients with acute ST-segment elevation myocardial infarction
Siyu PEI ; Hao MIAO ; Xin LIU ; Chengwen CHEN ; Yang DUAN ; Min ZHANG ; Yuan LU ; Wu XU ; Cheng WANG
Chinese Journal of Radiology 2023;57(8):897-903
Objective:To investigate the clinical application value of left ventricular myocardial strain obtained by cardiac MR (CMR) in recent major adverse cardiovascular events (MACE) in patients with acute ST-segment elevation myocardial infarction (STEMI) undergoing primary percutaneous coronary intervention (PCI).Methods:From January 2020 to December 2020, a total of 163 patients successfully underwent primary PCI and underwent CMR examination within one week after surgery at Affiliated Hospital of Xuzhou Medical University. The scan sequences included rapid balance-fast field echo and late-gadolinium enhancement. CVI42 post-processing software was used to analyze and measure the left ventricular myocardial strain indices, including left ventricular global longitudinal strain (GLS), left ventricular global circumferential strain (GCS), and left ventricular global radial strain (GRS). According to the results of the 1-year follow-up after surgery, the patients were divided into the MACE group ( n=28) and the non-MACE group ( n=135). For continuous variables with a normal distribution, the t test of two independent samples was used for comparisons between groups. For continuous variables with an abnormal distribution, the variables were compared and analyzed by the rank sum test. For categorical variables, the χ 2 tests were used for between-group comparisons. Cox regression was used to analyze the prognostic value of myocardial strain on the development of MACE in patients with STEMI. The receiver operating characteristic (ROC) curve was used to analyze the diagnostic efficacy of myocardial strain parameters, and the optimal cut-off value was evaluated by calculating the Youden index. Results:The GLS, GCS, and GRS of the MACE group were (-10.4±3.3)%, [-11.9 (-14.5, -9.3)]%, and (18.3±6.3)%, respectively, and those of the non-MACE group were (-13.7±3.4)%, [-14.6 (-16.4, -11.7)]%, and (22.3±6.1)%, respectively. The difference between the two groups was statistically significant ( t/ Z=-4.71, -3.04, 3.21, P<0.05). Multivariate Cox regression analysis showed that GLS was an independent predictor of MACE (HR=1.546, 95%CI 1.180-2.027, P=0.002). The ROC curve analysis showed that GLS had the largest area under the curve (AUC) (AUC=0.754, 95%CI 0.658-0.851, P<0.001), with a cut-off value of -12.45%. Its diagnostic sensitivity was 71.4%, and the specificity was 67.4%. The value was better than that of the traditional predictor of STEMI prognosis, namely, left ventricular ejection fraction (AUC=0.680, 95%CI 0.567-0.793, P=0.003). Conclusion:GLS of CMR is an independent predictor of MACE in STEMI patients undergoing primary PCI.

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