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.Targeting ER lipid raft-associated 1 reveals a coordinated cholesterol-dependent vulnerability in hepatocellular carcinoma
Yiming ZHANG ; Yushan HOU ; Xinxin WANG ; Kaikun XU ; Pei JIANG ; Siqi WANG ; Huimin KANG ; Hu ZHANG ; Jingzhuo JIN ; Xiaofen HUANG ; Zifeng LIU ; Songpeng YANG ; Jiaqi LIU ; Lingqiang ZHANG ; Fuchu HE ; Chunyan TIAN ; Aihua SUN
Clinical and Molecular Hepatology 2026;32(2):866-883
Background/Aims:
Dysregulated cholesterol metabolism is a hallmark of hepatocellular carcinoma (HCC) that drives tumor initiation and progression. However, clinical targeting of cholesterol metabolism has yielded limited benefits due to stringent feedback in tumor cells. Identifying a central mediator capable of restoring cholesterol homeostasis within the cell’s intrinsically fine-tuned regulatory framework is urgently needed.
Methods:
We integrated a proteomic dataset from patients with cholesterol-dysregulated HCC into a global cholesterol metabolic regulatory network to identify potential therapeutic targets for disrupted cholesterol homeostasis. The prognostic significance of the candidate targets was further validated in an independent cohort through immunohistochemistry. Functional and mechanistic studies were conducted in vitro using HCC cell lines and in vivo using mouse models. The pharmacological efficacy of the candidate agent was evaluated in both subcutaneous and orthotopic HCC mouse models.
Results:
ER lipid raft-associated 1 (ERLIN1), a pivotal regulator of cholesterol metabolism reprogramming, was identified as an independent favorable prognostic indicator in HCC. ERLIN1 constrains HCC progression both in vitro and in vivo by stabilizing the INSIG1–SCAP–SREBP2 axis and maintaining the metabolic balance of intracellular cholesterol. Under hypoxia, impaired factor-inhibiting hypoxia-1-dependent hydroxylation of ASB11 at asparagine residues 90 and 92 enhances ASB11-mediated ERLIN1 degradation. Pharmacological targeting of this axis using zoledronic acid (ZoA) attenuated HCC progression by weakening the ASB11–ERLIN1 interaction and restoring cholesterol homeostasis.
Conclusions
ERLIN1 represents a druggable metabolic vulnerability in cholesterol-dysregulated HCC. Targeting the ASB11–ERLIN1 axis with the clinically approved ZoA reestablishes cholesterol homeostasis and offers a promising therapeutic strategy to overcome the current limitations of cholesterol-targeted HCC therapies.
3.Risk Assessment for Ramadan Fasting in People With Diabetes in Hospital-Based Diabetes Clinics Using the Updated 2026 IDF-DAR Risk Calculator
Raja Nurazni Raja Azwan ; Chin Voon Tong ; Lisa Mohamed Nor ; Marisa Khatijah Borhan ; Syarifah Syahirah Syed Abas ; Poh Shean Wong ; Ying Jie Tan ; Shartiyah Ismail ; Eunice Yi Chwen Lau ; Yueh Chien Kuan ; Noor Hafis Md Tob ; Shu Teng Chai ; Pei Lin Chan ; Xe Hui Lee ; Wei Wei Ng ; Jin Hui Ho ; Miza Hiryanti Zakaria ; Rabeah Md Zuki ; Wan Mohd Hafez Wan Hamzah ; Melissa Vergis ; Choon Peng Sun ; Vanusha Devaraja Pillai ; Chee Koon Low ; Shazatul Reza Mohd Redzuan ; Xin-Yi Ooi ; Siti Sanaa Wan Azman ; Deviga Lachumanan ; Saiful Shahrizal Shudim ; Zanariah Hussein
Journal of the ASEAN Federation of Endocrine Societies 2026;41(S1):42-43
Introduction:
The 2021 IDF-DAR risk calculator had been previously
evaluated in multiple studies and subsequently widely
accepted and applied in clinical practice as a practical
standardized tool for patient risk stratification. Recently
updated, the 2026 IDF-DAR Risk calculator enables a more individualized, evidence-related evaluation of patientrelated and disease-related risk factors, incorporating
modern diabetes technologies, including continuous
glucose monitoring (CGM), automated insulin delivery
(AID) systems, and advanced insulin formulations to
enhance risk stratification. This tool allows medical
professionals to tailor Ramadan practices based on overall
factors toward promoting safe fasting.
Methodology:
This prospective multicentre observational study recruited
adults with Type 1 and Type 2 diabetes attending public
hospitals nationwide. People with diabetes (PwD) intending
to perform Ramadan fasting were invited to participate
and assessed using the 2026 IDF-DAR Risk Calculator in
the 6-week pre-Ramadan period between 30th January and
19th March 2026.
Results:
A total of 458 PwD were evaluated and stratified into low
(15.7%), moderate (41%), and high risk (43.3%) categories.
Most participants had Type 2 diabetes (83.6%), with 60.3%
having a disease duration exceeding 10 years and 43%
exhibiting poor glycemic control (hemoglobin A1c >9%).
Insulin therapy was used by 76.4% of participants, including
two individuals with Type 1 diabetes using AID systems.
Most participants reported no recent hypoglycemia (76.4%),
81.0% performed glucose monitoring, and 3.3% used CGM.
Severe comorbidities were uncommon, with 1.1% having
unstable macrovascular disease and 4.4% advanced chronic
kidney disease (estimated glomerular filtration rate <30).
Notably, 72.2% received structured Ramadan education.
Conclusion
Majority of PwD attending tertiary diabetes clinics were
in the moderate- to high-risk category and intended to
fast despite medical advice against fasting in some cases.
Although most participants were on insulin therapy,
hypoglycemia was low in the pre-Ramadan period.
Integration of modern technologies, advanced insulin
therapies, and structured education may support safer
fasting practices.
Risk Assessment
;
Diabetes Mellitus
;
Hospitals
;
Fasting
4.A clinical study on the prognostic value of preoperative prognostic nutritional index in patients with renal cell carcinoma undergoing partial nephrectomy
Yishuai ZHANG ; Yanxin ZHUANG ; BAOERBIEKE YELINAER ; Ying WANG ; Chenhui MA ; Jin ZENG ; Xinqi PEI
Journal of Modern Urology 2026;31(5):428-434
Objective To investigate the clinical prognostic value of the prognostic nutritional index (PNI) in patients with renal cell carcinoma (RCC) undergoing partial nephrectomy.Methods A retrospective analysis was conducted on the clinical data of RCC patients who underwent partial nephrectomy at our hospital during 2020 and 2024.The optimal cutoff value of PNI was determined based on the maximum Youden index from the receiver operating characteristic (ROC) curve.The clinicopathological characteristics, overall survival (OS), and progression-free survival (PFS) between patients with different PNI levels were compared.Cox regression analysis was used to explore the independent prognostic factors for RCC patients. Results Based on the ROC curve, the optimal cutoff value corresponding to the maximum Youden index was calculated to be 46.675, at which point the sensitivity was 65.3% and the specificity was 62.2%.Of the 815 patients included in the study, 293 (36.0%) patients had a PNI<46.675.Cox regression analysis confirmed that preoperative PNI, tumor diameter, ISUP classification, and ECOG score were independent risk factors for postoperative OS in patients with RCC who underwent PN (P<0.05).In addition, preoperative PNI, tumor diameter, ISUP classification and ECOG score were independent risk factors for postoperative PFS (P<0.05).After covariates in the multivariable Cox regression model were adjusted, patients in the low PNI group demonstrated significantly shorter overall survival (HR=2.392, P<0.001) and significantly poorer progression-free survival (HR=2.076, P<0.001) compared to those in the high PNI group.Conclusion Preoperative PNI has significant clinical value in evaluating the prognosis of RCC patients undergoing partial nephrectomy.An elevated preoperative PNI is associated with a better prognosis.
5.MADS-Net: a Multi-scale Attention and Dynamic Sparse Mask Fusion Network for Electrical Impedance Tomography Image Reconstruction in Breast Screening
Cheng-Tao YU ; Jia-Hui WANG ; Song-Pei HU ; Huai-Jin CHEN ; Li DING ; Xia YE ; Jia-Feng YAO
Progress in Biochemistry and Biophysics 2026;53(9):2480-2496
ObjectiveTo address the key challenges in breast electrical impedance tomography (EIT) reconstruction including the severe ill‑posedness of the inverse problem, limited measurement information from the electrodes, low contrast of small lesions, as well as the common drawbacks of traditional reconstruction methods such as boundary blurring, low spatial resolution, and weak small-target detection capability, and to overcome the limitations of existing deep learning approaches namely low training efficiency, insufficient physical consistency, and substantial information loss during the low‑to‑high dimensional mapping process, this study proposes a multi‑scale attention and dynamic sparse mask fusion network (MADS‑Net). The proposed method aims to enhance high‑resolution reconstruction accuracy, small-lesion detectability, noise robustness, and computational efficiency in breast‑screening-oriented EIT, thereby providing a feasible solution for the application of EIT in breast functional imaging. MethodsThe proposed network adopts a strategy of “locate first, reconstruct later”. It realizes adaptive allocation of computational resources through a dynamic mask module, alleviates information loss in the low-to-high dimensional mapping process via a multi-scale attention mechanism, and embeds physical constraints to improve the physical rationality of reconstruction results. The performance of the proposed method was validated through systematic numerical simulations, agar phantom experiments, and porcine tissue experiments. Comparisons were made with mainstream methods including the Gauss-Newton algorithm, convolutional neural networks, U-Net, and Deep D-Bar. The image correlation coefficient (ICC), root mean square error (RMSE), and peak signal-to-noise ratio (PSNR) were adopted as quantitative evaluation metrics. Furthermore, lesion‑level evaluation metrics were introduced to assess the capability of different methods in reconstructing abnormal target boundaries and small‑sized target regions. ResultsExperimental results demonstrate that MADS-Net outperforms all comparison methods across all test scenarios. In numerical simulations, it achieves higher accuracy in target morphology and spatial location recovery as well as better background suppression. In agar phantom experiments, its detection and imaging reliability for small targets is significantly superior to the comparison methods. In porcine tissue experiments, the model maintains good generalization performance and can adapt to complex biological tissue imaging scenarios. Quantitative results show that MADS-Net achieves an average ICC of 0.917 6±0.048 8, an RMSE as low as0.060 6±0.031 6, and a PSNR of (23.893 9±2.685 3) dB. It successfully detects tiny targets as small as 5 mm within a 100 mm diameter sensing field, reduces the total training duration by 23.5% compared with the baseline method, and maintains stable reconstruction performance even under strong noise interference. ConclusionThe results demonstrate that MADS-Net achieves favorable EIT reconstruction performance in numerical simulations, agar phantom experiments, and porcine tissue experiments, even under common practical imaging challenges including low signal-to-noise ratio and blurred tissue boundaries. This study provides a methodological reference for further research on EIT in breast-screening-related scenarios and other medical functional imaging tasks, and also offers a low-cost, radiation-free alternative for breast lesion screening that aligns with the clinical demand for non-invasive diagnostic tools. It should be noted that real clinical breast data have not yet been used for validation in this study, and the clinical applicability of the proposed method requires further investigation. Future work will prioritize collecting multi-center real clinical breast datasets, and further optimize the model’s generalization ability to adapt to individual differences in breast tissue composition to promote its clinical application.
6.Hei Xiaoyaosan Ameliorates Cognitive Impairment in Rat Model of Alzheimer's Disease by Clearing Peripheral Aβ via AMPK/SREBP2/PCSK9/LRP-1 Signaling Pathway
Zifan NIE ; Yingchuan MA ; Wenli PEI ; Yumei HAN ; Jiao YANG ; Ruixue JIN ; Qian WANG ; Huping WANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(22):61-73
ObjectiveTo investigate the mechanism of Hei Xiaoyaosan (HXS) in ameliorating cognitive impairment in the rat model of Alzheimer's disease (AD) by regulating the adenosine 5'-monophosphate-activated protein kinase (AMPK)/sterol regulatory element-binding protein 2 (SREBP2)/proprotein convertase subtilisin/kexin type 9 (PCSK9)/low-density lipoprotein receptor-related protein 1 (LRP-1) signaling pathway to clear amyloid-β (Aβ) from the peripheral circulation. MethodsNinety 8-week-old specific pathogen-free (SPF)-grade male Sprague-Dawley (SD) rats were acclimatized for 1 week. After acclimatization, 10 rats were randomly selected as the blank control group, and the remaining 80 rats were prepared for surgical modeling. Among the 80 rats for modeling, 70 were anesthetized and injected with 1 μL of amyloid-β1-42 (Aβ1-42) solution into the bilateral hippocampal CA1 region for the modeling of AD, and the remaining 10 were assigned to the sham operation group, which received an injection of 1 μL normal saline at the same anatomical site. The successfully modeled rats were randomly assigned into 6 groups (n=10 per group): a model group, a donepezil hydrochloride group, an acadesine (AICAR, an AMPK agonist) group, and high-, medium-, and low-dose HXS groups. After 42 consecutive days of administration intervention, the Morris water maze test and T-maze test were performed to evaluate the learning and memory abilities of rats. Fluoro-Jade B (FJB) staining was conducted to observe pathological changes of neurons in the rat hippocampus, and thioflavin S staining to quantify the area of Aβ deposition in the hippocampal region. Immunofluorescence double staining (IF-DS) was performed to detect the expression of LRP-1 and Aβ signals in the liver tissue. Enzyme-linked immunosorbent assay (ELISA) was employed to measure the levels of Aβ1-42 and soluble LRP-1 (sLRP-1) in the serum. Western blot was performed to determine the protein levels of AMPK, phosphorylated AMPK (p-AMPK), SREBP2, PCSK9, and LRP-1 in the liver tissue. Real time polymerase chain reaction(Real-time PCR) was adopted to assess the mRNA levels of AMPK, SREBP2, PCSK9, and LRP-1 in the liver tissue. ResultsCompared with the blank control group, the model group showed increased total swimming distance on day 6 and path length in the target quadrant on day 7 (P<0.01), a decreased correct alternation rate in the T-maze test (P<0.01), disarrangement of hippocampal neurons, increased FJB-positive cells (P<0.01), enlarged area of Aβ deposition (P<0.01). reduced LRP-1 fluorescence signals and enhanced Aβ fluorescence signals in the liver tissue (P<0.01), elevated serum level of Aβ1-42, declined serum level of sLRP-1 (P<0.01), downregulated protein levels of p-AMPK and LRP-1 as well as mRNA levels of AMPK and LRP-1, and upregulated protein and mRNA levels of SREBP2 and PCSK9 in the liver tissue (P<0.01). Compared with the model group, the high-dose HXS group, donepezil hydrochloride group, and AICAR group exhibited reduced total swimming distance and target quadrant path length (P<0.05, P<0.01), an increased correct alternation rate in the T-maze test (P<0.05), decreased FJB-positive cells (P<0.01), reduced Aβ deposition (P<0.01) in the hippocampal tissue, enhanced LRP-1 signals and weakened Aβ signals in the liver tissue (P<0.01), a declined serum level of Aβ1-42 and an elevated serum level of sLRP-1 (P<0.01), upregulated protein levels of p-AMPK and LRP-1 as well as mRNA levels of AMPK and LRP-1, and downregulated protein and mRNA levels of SREBP2 and PCSK9 in the liver tissue (P<0.01). ConclusionHei Xiaoyaosan can regulate the AMPK/SREBP2/PCSK9/LRP-1 signaling pathway to enhance the clearance of peripheral Aβ by the liver tissue and reduce Aβ deposition in the brain, thereby improving the learning and memory abilities of AD rats.
7.Hei Xiaoyaosan Ameliorates Cognitive Impairment in Rat Model of Alzheimer's Disease by Activating Autophagy Through Regulating AMPK/ULK1 Signaling Pathway
Wenli PEI ; Yumei HAN ; Zifan NIE ; Yingchuan MA ; Ruixue JIN ; Qian WANG ; Jiao YANG ; Huping WANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(22):91-101
ObjectiveTo investigate the effect and mechanism of Hei Xiaoyaosan in preventing and treating Alzheimer's disease (AD) by activating autophagy through regulating the adenosine monophosphate-activated protein kinase (AMPK)/UNC-51 like autophagy activating kinase 1 (ULK1) signaling pathway. MethodsNinety 8-week-old male SPF-grade SD rats were selected and randomized into two groups: a blank control group (n=10) and a modeling group (n=80). From the modeling group, 10 rats were randomly assigned as a sham group (bilateral hippocampal injection of 1 μL normal saline), while the remaining 70 rats were injected with 1 μL Aβ1-42 solution bilaterally into the hippocampus to replicate the AD model. The Morris water maze test was conducted to screen the successfully modeled rats, which were then randomly allocated to the model, donepezil hydrochloride, 5-aminoimidazole-4-carboxamide 1-β-D-ribofuranoside (AICAR, an AMPK agonist), and high-, medium-, and low-dose Hei Xiaoyaosan groups, with 10 rats per group. After 42 days of drug intervention, the Morris water maze and open field tests were conducted to assess learning and memory abilities and motor function. Pathological changes in the hippocampal tissue were observed by Nissl staining, and autophagy-related pathological damage was examined via transmission electron microscopy (TEM). The expression of LC3 in the hippocampus was observed by immunofluorescence staining. The protein levels of AMPK, phosphorylate (p)-AMPK, ULK1, p-ULK1, p62, and autophagy related 7 (Atg7) were determined by Western blot, while the mRNA levels of AMPK and ULK1 were measured by Real-time polymerase chain reaction (Real-time PCR). ResultsCompared with the blank control group, the model group showed increased escape latency (P<0.01) and decreased platform crossings (P<0.01) on day 6 in the water maze test, reduced total distance traveled and average speed in the open field test (P<0.01), disarranged neurons with Nissl body loss and evident autophagic damage, weakened LC3 fluorescence signal (P<0.01), downregulated protein levels of p-AMPK, p-ULK1, and Atg7 and mRNA levels of AMPK and ULK1 in the hippocampus (P<0.01), and upregulated protein level of p62 (P<0.01). Compared with the model group, the positive drug group, high-dose Hei Xiaoyaosan group, and AICAR group showed decreased escape latency (P<0.05, P<0.01) and increased platform crossings (P<0.01) on day 6, and increased total distance traveled and average speed in the open field test (P<0.05). The positive drug group, high- and medium-dose Hei Xiaoyaosan groups, and the AICAR group showed alleviated cellular structural damage, regular neuronal arrangement, increased Nissl bodies, and orderly distribution of autophagosomes. Furthermore, the LC3 fluorescence intensity was enhanced in the positive drug group, high- and medium-dose Hei Xiaoyaosan groups, and the AICAR group (P<0.01). The positive drug group, high-dose Hei Xiaoyaosan group, and the AICAR group exhibited upregulated protein and mRNA levels of p-AMPK and p-ULK1 (P<0.05, P<0.01), upregulated protein level of Atg7 (P<0.05, P<0.01), and downregulated protein level of p62 (P<0.01). ConclusionHei Xiaoyaosan can improve learning and memory abilities and alleviate pathological status in the rat model of AD by regulating the AMPK/ULK1 signaling pathway, enhancing cellular autophagy, and repairing damaged nerve cells.
8.Application of Carbon Nanoparticle-Manganese Dioxide Injection in Gastrointestinal Staining-Based Marking and Localization in Rats and Pigs
Wanlin CHEN ; Qingrui DONG ; Mengtao LI ; Nan LIN ; Hu ZHAO ; Zaizhong ZHANG ; Lingjun ZENG ; Pei LI ; Jin YANG ; Yu WANG ; Aiwen HUANG
Laboratory Animal and Comparative Medicine 2026;46(5):727-739
ObjectiveTo develop a carbon nanoparticle-manganese dioxide (CN-MnO2) injection integrating visually identifiable staining and magnetic resonance (MR) T1/T2 dual-modal imaging, and to systematically evaluate its physicochemical properties, critical quality attributes, biocompatibility, and efficacy for gastrointestinal marking and localization. MethodsCN-MnO2 was prepared using a mild liquid-phase in situ redox deposition strategy and characterized by transmission electron microscopy (TEM), dynamic light scattering (DLS), zeta potential analysis, X-ray photoelectron spectroscopy (XPS), and X-ray diffraction (XRD). The pH of CN-MnO2 was directly measured using a pH meter. The osmolality pressure of CN-MnO2 was determined by the freezing point depression method. The bacterial endotoxin was detected using the gel-clot Limulus amebocyte lysate test, which did not rely on absorbance readings. The appearance of the CN-MnO2 injection solution stored at 4 ℃ under light shielding and sealed for 0, 5, and 10 days was observed, and its hydrated particle size, polydispersity index, zeta potential, and pH were also measured. With gadopentetate dimeglumine (Gd-DTPA) as the positive control, different concentration gradients were established, and T1 and T2 relaxation times were measured to construct concentration-dependent plots of the longitudinal and transverse relaxation rates (R1 and R2) and calculate the corresponding relaxivities (r1 and r2), compare relaxivity performance, and determine the working concentration for subsequent in vivo experiments. Cytocompatibility was assessed using the methyl thiazolyl tetrazolium (MTT) assay and live/dead staining. Sixteen male 6-week-old SD rats were randomly assigned to an experimental group and a control group (n=8 per group) and subcutaneously injected with CN-MnO2 injection at a total Mn concentration of 0.80 mmol/L or an equal volume of normal saline, respectively. Short-term in vivo safety was evaluated by regular body weight monitoring, hematoxylin-eosin (HE) staining of major organs, hematological and serum biochemical analyses, inflammatory cytokine detection, and inductively coupled plasma optical emission spectrometry (ICP-OES) analysis. Eight male 5–6-month-old Bama minipigs were randomly assigned to a CN-MnO2 group (n=6) and a carbon nanoparticle suspension injection (CNSI) group (n=2). CN-MnO2 injection or CNSI was injected subserosally into the stomach, small intestine, and colon. Within 5 min after each injection, the maximum staining diameter at each injection site was measured and recorded. MR T1WI/T2WI scans were performed on days 3 and 7 after injection to evaluate MR visualization at the injection sites. At the end of observation, the injection sites and major organs were collected for histopathological evaluation by hematoxylin-eosin staining. ResultsCN-MnO2 appeared as a black suspension, with no obvious aggregation or sedimentation after standing at room temperature (20-25 ℃) for 15 days. Its average hydrodynamic diameter was approximately (228.7±15.3) nm, the polydispersity index was 0.184±0.021, and the zeta potential was (-24.6±2.8) mV. XPS showed that CN-MnO2 was mainly composed of C, O, and Mn, with oxygen accounting for approximately 68.82%. The Mn 2p spectrum exhibited two characteristic peaks at 642.4 and 653.1 eV. XRD indicated that low-crystallinity δ-MnO2 was dispersed within the carbon matrix. The pH was 6.82±0.06, the osmolality was (298.4±6.7) mOsm/kg, and the bacterial endotoxin level was below the detection limit. In vitro MR imaging showed a clear concentration-dependent response, with r1 and r2 values of 9.6 L·mmol-1·s-1 and 17.2 L·mmol-1·s-1, respectively, indicating favorable T1/T2 contrast performance superior to that of Gd-DTPA. At 0.80 mmol/L, CN-MnO2 showed no obvious cytotoxicity in vitro. Short-term safety evaluation in SD rats revealed no obvious inflammatory response, impairment of liver or renal function, or abnormal manganese accumulation in major organs. In Bama minipigs, the mean staining diameters of the CN-MnO2 group in the stomach, small intestine, and colon were (0.65±0.12) cm, (0.61±0.11) cm, and (0.58±0.10) cm, respectively, with no significant difference among the three injection sites (P>0.05). The overall mean staining diameter was smaller in the CN-MnO2 group than in the CNSI group (P<0.05). MR examinations showed that the injection sites were identifiable on both day 3 and day 7, with stronger signals observed on day 3. At the end of observation, no obvious pathological abnormalities were observed at the injection sites or in major organs. ConclusionA CN-MnO2 injection with uniform particle size and stable physicochemical properties can be successfully prepared. This formulation integrates visible gastrointestinal staining with MR T1/T2 dual-modal imaging, providing a new strategy for preoperative gastrointestinal marking and localization as well as precise intraoperative resection.
9.Correlation Analysis Between Microbial Community Changes and Medicinal Quality Formation During Processing of Angelicae Dahuricae Radix
Xiaoyan CHEN ; Xinglong ZHU ; Qingxia GAN ; Jiahao WANG ; Guangqin AN ; Qinghua WU ; Jin PEI ; Yuntong MA
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(4):198-207
ObjectiveTo compare the differences in color, odor, coumarin content and microbial community composition of Angelicae Dahuricae Radix(ADR) during different drying processes, and to explore the correlation between changes in microbial community composition and changes in quality indexes of ADR. MethodsThe fresh ADR was processed at three drying temperatures(50, 70, 100 ℃) by drying and steaming cutting, semi-fresh cutting and drying, fresh cutting and drying, and sulfur fumigation methods. The color values of samples were extracted by Adobe Photoshop 2022 software and subjected to principal component analysis(PCA), electronic nose was used to identify the odor information of medicinal powders and subjected to loadings analysis, PCA, and linear discriminant analysis(LDA), and high performance liquid chromatography(HPLC) was used to determine the contents of five coumarins(bergapten, oxypeucedanin, imperatorin, phellopterin, isoimperatorin). The samples for microbial detection were taken from fresh dried samples, 50 ℃(dried and steamed cut, sulfur fumigated) samples, and 100 ℃(dried and steamed cut) samples when the water content was 50% and 14%, respectively. And the changes of microbial community composition during processing were determined by high-throughput sequencing method. The relationship between the changes of microbial community composition and the changes of odor, color and active component content of ADR during drying process was analyzed by Pearson correlation analysis. ResultsThe color quantification results showed that an increase in drying temperature led to the decrease of brightness value(L), and the increases of red-green value(a) and yellow-blue value(b), and the change of processing method had no obvious effect on the color of medicinal materials. The results of odor quantification showed that W1S, W2S, W5S, W2W and W1W sensor were sensitive to the odor changes of ADR and could be used to distinguish ADR decoction pieces from different processing methods. The results of HPLC showed that the coumarin content of ADR decreased with the increase of drying temperature and the delay of processing time, the optimal processing method was drying and steaming cutting method, and the optimal temperature was 50 ℃. High-throughput sequencing results showed that the dominant bacteria in ADR during processing were Achromobacter, Agrobacterium, Nocardioides, Mycobacterium and Enterobacter, the dominant fungi were Coprinopsis, Meyerozyma and Apiotrichum. The results of correlation analysis showed that the quality indexes of ADR were positively correlated with Agrobacterium, Mycobacterium in bacteria, Candida in fungi, and negatively correlated with Bacillus in bacteria. ConclusionThere are significant differences in the color, odor, coumarin content and microbial community composition of ADR in different drying processes, and the best drying method is drying and steaming cutting at 50 ℃. The relative abundance changes of 9 bacterial genera and 4 fungal genera are closely related to the quality formation of ADR during the drying process.
10.Construction and Application of"On-Off-On"Fluorescence Sensor for Chlorpromazine Hydrochloride Based on Near Infrared Carbon Quantum Dots
Yu LIN ; Feng TAN ; Yu-Hua SHEN ; Li-Qin ZHU ; Pei-Yao YAN ; Jin-Tao PAN ; Kai-Shun LIU
Chinese Journal of Analytical Chemistry 2025;53(6):934-943
In this work,near infrared carbon quantum dots(NIR-CDs)were synthesized by hydrothermal method using biomass material Clausena lansium leaves.The synthesized NIR-CDs emitted maximum fluorescence signal at 677 nm,which was independent of excitation wavelength.The characterization results showed that there were abundant groups on the surface of NIR-CDs.Pd2+could form non-fluorescent compounds with the surface groups of NIR-CDs,resulting in fluorescence quenching(Fluorescence signal was denoted as F0).Because chlorpromazine hydrochloride(CPZ)parent nucleus contained unoxidized S atom,CPZ could form stable colored complex with Pd2+under acidic conditions.In the presence of CPZ,Pd2+dissociated from the surface of NIR-CDs and bonded with CPZ,so that the fluorescence signal could be restored(Fluorescence signal was denoted as F).An"on-off-on"fluorescence sensor was thus constructed.The fluorescence signal recovery value of NIR-CDs(△F=F-F0)showed a good linear relationship with the concentration of CPZ in the range of 5.68-28.43 μg/mL,and the detection limit(3σ)was 0.078 μg/mL.The sensor was applied to determination of CPZ in pharmaceutical preparations,and the recoveries were 94%-106%.The developed fluorescence sensor was expected to be used in quality control of actual pharmaceutical preparations.


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