1.Exploring Chemical Constituent Distribution in Blood/Brain(Hippocampus) and Emotional Regulatory Effect of Raw and Vinegar-processed Products of Citri Reticulatae Pericarpium Viride
Yi BAO ; Yonggui SONG ; Qianmin LI ; Zhifu AI ; Genhua ZHU ; Ming YANG ; Huanhua XU ; Qin ZHENG ; Yiting HUANG ; Zihan GAO ; Dan SU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(2):189-197
ObjectiveTo investigate the migration and distribution characteristics of chemical constituents in blood and hippocampal tissues before and after vinegar processing of Citri Reticulatae Pericarpium Viride(CRPV), and to explore the potential material basis and mechanisms underlying their regulatory effects on emotional disorders by comparing the effects of raw and vinegar-processed products of CRPV. MethodsUltra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS/MS) was employed to characterize and identify the chemical constituents of raw and vinegar-processed products of CRPV extracts, as well as their migrating components in blood and hippocampal tissues after oral administration. Reference standards, databases, and relevant literature were utilized for compound annotation, with data processing performed using PeakView 1.2 software. Seventy male C57BL/6 mice were randomly divided into seven groups, including the blank group, model group, diazepam group(2.5 mg·kg-1), raw CRPV low/high dose groups(0.6, 1.2 g·kg-1), and vinegar-processed CRPV low/high dose groups(0.6, 1.2 g·kg-1), with 10 mice per group. Except for the blank group, all other groups underwent chronic restraint stress(2 h·d-1) for 20 d. Each drug-treated group received oral administration at the predetermined dose starting 10 d after modeling, with a total treatment duration of 10 d. Following model-based drug administration, mice underwent open-field, forced swimming, and elevated plus maze tests. After anesthesia with isoflurane, whole brains were collected from each group of mice, and hippocampi were dissected. Reactive oxygen species(ROS) level in hippocampal tissues was quantified by enzyme-linked immunosorbent assay(ELISA). Hematoxylin-eosin(HE) staining was used to observe hippocampal tissue morphology. Immunofluorescence was performed to detect neuronal nuclei(NeuN) and peroxisome proliferator-activated receptor alpha(PPARα) expressions in hippocampal tissue. Then, pharmacodynamic evaluations were conducted to assess the effects of raw and vinegar-processed CRPV on mood disorders, exploring the potential mechanisms. ResultsVinegar processing caused significant changes in the chemical composition of CRPV, with 18 components showing increased relative content and 35 components showing decreased relative content. The primary changes occurred in flavonoid compounds, including 20 flavonoids, 20 flavonoid glycosides, 3 triterpenes, 3 phenolic acids, 1 alkaloid, and 6 other compounds. Twenty-one components were detected in blood(15 methoxyflavones, 4 flavonoid glycosides, and 2 phenolic acids), with 17 shared between raw and vinegar-processed CRPV. Seven components reached hippocampal tissues(all common to both forms). In regulating emotional disorders, Vinegar-processed CRPV exhibited superior antidepressant-like effects compared to raw products. HE staining revealed that both treatments improved hippocampal neuronal morphology, particularly in the damaged CA1 and CA3 regions. Immunofluorescence and ELISA analyses demonstrated that both raw and vinegar-processed CRPV significantly modulated NeuN and PPARα expressions in hippocampal tissue while alleviating oxidative stress induced by excessive ROS(P<0.05). ConclusionThe chemical composition of CRPV undergoes changes after vinegar processing, but the migrating components in blood and hippocampus are primarily methoxyflavonoids. These components may serve as the potential material basis for activating the PPARα pathway, thereby negatively regulating ROS generation in the hippocampus, reducing oxidative stress, and promoting the development of NeuN-positive neurons. These findings provide experimental evidence for enhancing quality standards, pharmacodynamic material research, and active drug development of raw and vinegar-processed CRPV.
2.Preliminary application of sacral neuromodulation in patients with benign prostatic hyperplasia complicated with underactive bladder after transurethral resection of the prostate
Ning LIU ; Yan ZHANG ; Tao LI ; Qiang HU ; Kai LU ; Lei ZHANG ; Jianping WU ; Shuqiu CHEN ; Bin XU ; Ming CHEN
Journal of Modern Urology 2025;30(1):39-42
[Objective] To evaluate the efficacy and safety of sacral neuromodulation (SNM) in the treatment of patients with benign prostatic hyperplasia (BPH) complicated with underactive bladder (UAB) who respond poorly to transurethral resection of the prostate (TURP). [Methods] A retrospective analysis was performed on 10 patients with BPH and UAB treated with TURP by the same surgeon in Zhongda Hospital Southeast University during Jan.2018 and Jan.2023.The residual urine volume was not significantly relieved after operation, and the maximum urine flow rate and urine volume per discharge were not significantly improved.All patients underwent phase I SNM, and urinary diaries were recorded before and after surgery to observe the average daily frequency of urination, volume per urination, maximum urine flow rate, and residual urine volume. [Results] The operation time was (97.6±11.2) min.During the postoperative test of 2-4 weeks, if the residual urine volume reduction by more than 50% was deemed as effective, SNM was effective in 6 patients (60.0%). Compared with preoperative results, the daily frequency of urination [(20.2±3.8) times vs. (13.2±3.2) times], volume per urination [(119.2±56.7) mL vs. (246.5±59.2) mL], maximum urine flow rate [(8.7±1.5) mL/s vs. (16.5±2.6) mL/s], and residual urine volume [(222.5±55.0) mL vs. (80.8±16.0) mL] were significantly improved, with statistical significance (P<0.05). There were no complications such as bleeding, infection, fever or pain.The 6 patients who had effective outcomes successfully completed phase II surgery, and the fistula was removed.During the follow-up of 1 year, the curative effect was stable, and there were no complications such as electrode displacement, incision infection, or pain in the irritation sites.The residual urine volume of the other 4 unsuccessful patients did not improve significantly, and the electrodes were removed and the vesicostomy tube was retained. [Conclusion] SNM is safe and effective in the treatment of BPH with UAB patients with poor curative effects after TURP.
3.Effect of Icariin on Steroid-induced Ferroptosis in Rat Bone Microvascular Endothelial Cells
Jiancheng TANG ; Yue ZHANG ; Ruichen JIANG ; Zhengrong YUE ; Ming LI ; Yaqi ZHANG ; Zetao YIN ; Weiguo WANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(5):131-140
ObjectiveTo investigate the effect of icariin (ICA) on steroid-induced ferroptosis in bone microvascular endothelial cells (BMECs). MethodsRat BMECs were selected and treated with 500 mg·L-1 hydrocortisone for 1.5 h to establish a ferroptosis model of BMECs. The experimental cells were divided into a blank group, hormone group (500 mg·L-1 hydrocortisone), ICA group (500 mg·L-1 hydrocortisone + 34 mg·L-1 ICA), and ferroptosis agonist group (500 mg·L-1 hydrocortisone + 34 mg·L-1 ICA + 2.7 mg·L-1 erastin). Cell viability was detected by CCK-8. The levels of ferrous ion, glutathione (GSH), malondialdehyde (MDA), superoxide dismutase (SOD), and reactive oxygen species (ROS) were detected by related kit species. The ferroptosis-related proteins, such as glutathione peroxidase 4(GPX4), ferritin light chain (FTL), and transferrin receptor protein1 (sTfR) were detected by Western blot, as well as autophagy-related proteins including microtubule-associated protein 1 light chain 3B (LC3B), Beclin1, B-cell lymphoma-2 (Bcl-2), and Caspase-3. Results500 mg·L-1 hydrocortisone intervention for 1.5 h could effectively induce ferroptosis in BMECs, and ferroptosis levels could reach a peak as the intervention continued. In terms of cellular antioxidant capacity, compared with those in the blank group, the cell vitality, GSH in the hormone group decreased significantly, and the levels of ROS, SOD, MDA, and ferrous ions were significantly increased (P<0.01). Compared with those in the hormone group, the cell viability, GSH were significantly increased, and the levels of ROS, SOD, MDA, and ferrous ions were decreased in the ICA group (P<0.01). Compared with those in the ICA group, the cell vitality, GSH in the ferroptosis agonist group decreased significantly, and the levels of ROS, SOD, MDA, and ferrous ions increased significantly (P<0.01). In terms of the relationship between ferroptosis and autophagy, compared with the blank group, the hormone group had significantly increased expression levels of LC3B, sTfR, Beclin1, and FTL and significantly decreased expression levels of GPX4 (P<0.01). Compared with the hormone group, The ICA group had significantly decreased expression levels of LC3B, sTfR, and FTL and significantly increased expression levels of Beclin 1 and GPX4 (P<0.01). Compared with those in the ICA group, the expression levels of LC3B, sTfR, and FTL increased in the rapamycin group, and those of Beclin 1 and GPX4 decreased (P<0.01). In terms of cell ferroptosis and apoptosis,compared with the blank group, the hormone group had significantly increased expression levels of FTL, sTfR and Caspase-3 and significantly decreased expression levels of GPX4, and Bcl-2 (P<0.01). Compared with the hormone group, the ICA group had significantly decreased expression levels of FTL, sTfR and Caspase-3 and significantly increased expression levels of GPX4, and Bcl-2 (P<0.01). Compared with those in the ICA group, the expression levels of FTL, sTfR and Caspase-3 in the ferroptosis agonist group were increased, and the expression levels of GPX4, and Bcl-2 were decreased (P<0.01). In terms of cell function,compared with that in the blank group, the ability of cell migration and tube formation was significantly decreased in the hormone group (P<0.01). Compared with that in the hormone group, the cell migration and tube formation ability in the ICA group were significantly increased (P<0.01). ConclusionFerroptosis is involved in steroid-induced damage in BMECs. ICA can inhibit steroid-induced ferroptosis in BMECs, and the mechanism may be associated with the inhibition of ferroptosis by regulating autophagy.
4.Effects of clopidogrel on the pharmacokinetics and pharmacodynamics of ciprofol in rats
Ming LU ; Xiaoyu YIN ; Wenli LI ; Shan LI ; Xiangchen LI ; Zhiqing ZHANG
China Pharmacy 2025;36(2):179-184
OBJECTIVE To investigate the effects of clopidogrel on the pharmacokinetics and pharmacodynamics of ciprofol in rats. METHODS Eighteen male SD rats were randomly divided into control group, clopidogrel normal-dose group and clopidogrel high-dose group, with 6 rats in each group. Among them, rats in the normal-dose group and high-dose group were given 7.5 mg/kg and 15 mg/kg clopidogrel by gavage, respectively, and rats in the control group were given the same volume of 0.5% sodium carboxymethyl cellulose solution, once a day, for 14 consecutive days. Afterward, 2.4 mg/kg ciprofol was injected by tailvein and blood samples were collected from the inner canthus of the eye at 2, 4, 8, 12, 16, 20, 30, 45 and 60 min after the end of the administration. During this period, the duration of the loss of righting reflex (LORR) in rats was counted. After the proteins were precipitated by acetonitrile, the rat plasma sample was analyzed by LC-MS/MS using deuterated ciprofol as the internal standard, Symmetry C18 as the chromatographic column, and acetonitrile-0.01% ammonia solution containing 5 mmol/L ammonium acetate (gradient elution) as the mobile phase to detect the concentration of ciprofol in the plasma. The pharmacokinetic parameters in rats were calculated by using DAS 2.0 software. RESULTS Compared with control group, area under the drug concentration-time curve and mean residence time of ciprofol increased or prolonged significantly, while plasma clearance decreased significantly in clopidogrel normal-dose and high-dose groups; the duration of LORR in rats was prolonged by 19.5% and 23.9%, with statistical difference (P<0.05). However, there was no statistically significant difference in the pharmacokinetic parameters or LORR duration of ciprofol between the different dose groups of clopidogrel (P>0.05). CONCLUSIONS Clopidogrel could inhibit the metabolism of ciprofol in rats and prolong the duration of LORR.
5.Classification and advances in clinical research of artificial colloidal plasma substitutes
Zhengyang CHANG ; Ming LI ; Jianpeng GAO ; Jing ZHANG ; Hua LYU ; Licheng ZHANG
Chinese Journal of Blood Transfusion 2025;38(1):136-141
The number of patients with reduced blood volume due to haemorrhage, fractures, severe infections, extensive burns and tumours is increasing, and traditional blood products are no longer able to meet the increasing clinical demand. Therefore, plasma substitutes have become particularly important in fluid resuscitation, especially artificial colloidal solutions, which have a sustained volume expansion time and a good volume expansion effect, and can significantly improve the circulatory status of patients. This article aims to review the classification of artificial colloidal plasma substitutes and their research progress in clinical practice, in order provide a more rigorous, professional and standardized reference for medicine.
6.Effects of clopidogrel on the pharmacokinetics and pharmacodynamics of ciprofol in rats
Ming LU ; Xiaoyu YIN ; Wenli LI ; Shan LI ; Xiangchen LI ; Zhiqing ZHANG
China Pharmacy 2025;36(2):179-184
OBJECTIVE To investigate the effects of clopidogrel on the pharmacokinetics and pharmacodynamics of ciprofol in rats. METHODS Eighteen male SD rats were randomly divided into control group, clopidogrel normal-dose group and clopidogrel high-dose group, with 6 rats in each group. Among them, rats in the normal-dose group and high-dose group were given 7.5 mg/kg and 15 mg/kg clopidogrel by gavage, respectively, and rats in the control group were given the same volume of 0.5% sodium carboxymethyl cellulose solution, once a day, for 14 consecutive days. Afterward, 2.4 mg/kg ciprofol was injected by tailvein and blood samples were collected from the inner canthus of the eye at 2, 4, 8, 12, 16, 20, 30, 45 and 60 min after the end of the administration. During this period, the duration of the loss of righting reflex (LORR) in rats was counted. After the proteins were precipitated by acetonitrile, the rat plasma sample was analyzed by LC-MS/MS using deuterated ciprofol as the internal standard, Symmetry C18 as the chromatographic column, and acetonitrile-0.01% ammonia solution containing 5 mmol/L ammonium acetate (gradient elution) as the mobile phase to detect the concentration of ciprofol in the plasma. The pharmacokinetic parameters in rats were calculated by using DAS 2.0 software. RESULTS Compared with control group, area under the drug concentration-time curve and mean residence time of ciprofol increased or prolonged significantly, while plasma clearance decreased significantly in clopidogrel normal-dose and high-dose groups; the duration of LORR in rats was prolonged by 19.5% and 23.9%, with statistical difference (P<0.05). However, there was no statistically significant difference in the pharmacokinetic parameters or LORR duration of ciprofol between the different dose groups of clopidogrel (P>0.05). CONCLUSIONS Clopidogrel could inhibit the metabolism of ciprofol in rats and prolong the duration of LORR.
7.Research advances in liver venous deprivation
Bensong HE ; Ming XIAO ; Qijia ZHANG ; Canhong XIANG ; Yanxiong WANG ; Yingbo LI ; Zhishuo WANG
Journal of Clinical Hepatology 2025;41(1):183-188
Portal vein embolization (PVE) can induce atrophy of the embolized lobe and compensatory regeneration of the non-embolized lobe. However, due to inadequate regeneration of future liver remnant (FLR) after PVE, some patients remain unsuitable for hepatectomy after PVE. In recent years, liver venous deprivation (LVD), which combines PVE with hepatic vein embolization (HVE), has induced enhanced FLR regeneration. Compared with associating liver partition and portal vein ligation for staged hepatectomy (ALPPS), LVD triggers faster and more robust FLR regeneration, with lower incidence rate of postoperative complications and mortality rate. By reviewing related articles on LVD, this article introduces the effectiveness of LVD and analyzes the differences and safety of various technical paths, and it is believed that LVD is a safe and effective preoperative pretreatment method.
8.Research and Application of Scalp Surface Laplacian Technique
Rui-Xin LUO ; Si-Ying GUO ; Xin-Yi LI ; Yu-He ZHAO ; Chun-Hou ZHENG ; Min-Peng XU ; Dong MING
Progress in Biochemistry and Biophysics 2025;52(2):425-438
Electroencephalogram (EEG) is a non-invasive, high temporal-resolution technique for monitoring brain activity. However, affected by the volume conduction effect, EEG has a low spatial resolution and is difficult to locate brain neuronal activity precisely. The surface Laplacian (SL) technique obtains the Laplacian EEG (LEEG) by estimating the second-order spatial derivative of the scalp potential. LEEG can reflect the radial current activity under the scalp, with positive values indicating current flow from the brain to the scalp (“source”) and negative values indicating current flow from the scalp to the brain (“sink”). It attenuates signals from volume conduction, effectively improving the spatial resolution of EEG, and is expected to contribute to breakthroughs in neural engineering. This paper provides a systematic overview of the principles and development of SL technology. Currently, there are two implementation paths for SL technology: current source density algorithms (CSD) and concentric ring electrodes (CRE). CSD performs the Laplace transform of the EEG signals acquired by conventional disc electrodes to indirectly estimate the LEEG. It can be mainly classified into local methods, global methods, and realistic Laplacian methods. The global method is the most commonly used approach in CSD, which can achieve more accurate estimation compared with the local method, and it does not require additional imaging equipment compared with the realistic Laplacian method. CRE employs new concentric ring electrodes instead of the traditional disc electrodes, and measures the LEEG directly by differential acquisition of the multi-ring signals. Depending on the structure, it can be divided into bipolar CRE, quasi-bipolar CRE, tripolar CRE, and multi-pole CRE. The tripolar CRE is widely used due to its optimal detection performance. While ensuring the quality of signal acquisition, the complexity of its preamplifier is relatively acceptable. Here, this paper introduces the study of the SL technique in resting rhythms, visual-related potentials, movement-related potentials, and sensorimotor rhythms. These studies demonstrate that SL technology can improve signal quality and enhance signal characteristics, confirming its potential applications in neuroscientific research, disease diagnosis, visual pathway detection, and brain-computer interfaces. CSD is frequently utilized in applications such as neuroscientific research and disease detection, where high-precision estimation of LEEG is required. And CRE tends to be used in brain-computer interfaces, that have stringent requirements for real-time data processing. Finally, this paper summarizes the strengths and weaknesses of SL technology and envisages its future development. SL technology boasts advantages such as reference independence, high spatial resolution, high temporal resolution, enhanced source connectivity analysis, and noise suppression. However, it also has shortcomings that can be further improved. Theoretically, simulation experiments should be conducted to investigate the theoretical characteristics of SL technology. For CSD methods, the algorithm needs to be optimized to improve the precision of LEEG estimation, reduce dependence on the number of channels, and decrease computational complexity and time consumption. For CRE methods, the electrodes need to be designed with appropriate structures and sizes, and the low-noise, high common-mode rejection ratio preamplifier should be developed. We hope that this paper can promote the in-depth research and wide application of SL technology.
9.Extraction and identification of primary rat brain microvascular endothelial cells by improved tissue block culture method
Fan Zhang ; Bolin Li ; Ming Chi ; Haiqin Liu ; Yuanyu Tang
Acta Universitatis Medicinalis Anhui 2025;60(1):10-14
Objective :
To investigate the brain microvascular tissue block culture method for extracting primary rat brain microvascular endothelial cells and identify its effect.
Methods :
Brain tissue from 4-week-old Sprague Dawley rats was screened, pre-digested and solidified to obtain brain microvascular segments. These segments were subsequently placed in a CO2incubator for primary culture. The target cells were identified by cell morphology and immunocytochemical staining for factor Ⅷ-related antigen.
Results :
After a 48-hour culture periodin vitro, the short spindle cells crawled out from around the brain microvascular segments. After 72 hours, island-like cell culsters formed. After 96 hours the clusters fused and the cells formed a typical monolayer, cobble stone-like, and mosaic arrangement. Factor Ⅷ-related antigen immunocytochemical staining showed that the cytoplasm of the cells appeared brown-red, indicating positive expression; DAB stained the nucleus, showing blue-dark.
Conclusion
The brain microvascular tissue block culture method can isolate and culture primary rat brain microvascular endothelial cells.
10.Protective effect of Shenfu injection against neonatal hypoxic-ischemic brain injury by inhibiting the ferroptosis
Xiaotong Zhang ; Meng Zhang ; Gang Li ; Yang Hu ; Yajing Xun ; Hui Ding ; Donglin Shen ; Ming Wu
Acta Universitatis Medicinalis Anhui 2025;60(1):31-40
Objective :
To observe the brain tissue injury during hypoxia-ischemia, as well as the pathological changes and the expression of ferroptosis-related factors after the use of Shenfu injection(SFI), and to explore the protective effect of SFI on hypoxic-ischemic brain injury(HIBD) by inhibiting ferroptosis.
Methods :
An animal model of HIBD in SD rats was constructed and intervened with SFI. Pathologic changes in brain tissue were observed by HE staining methods. Nissen staining was used to observe neuron survival. Glutathione Peroxidase 4(GPX4) and Divalent Metal Transporter 1(DMT1) expression were detected in brain tissue by Western blot, immunohistochemistry and immunofluorescence. Reduced Glutathione(GSH), Lactate Dehydrogenase(LDH), Malondialdehyde(MDA), Superoxide Dismutase(SOD) and tissue iron content were determined with the kits. BV-2 microglial cell line(BV2) cells were culturedin vitroand divided into control group(Ctrl group), oxygen-glucose deprivation group(OGD group), iron ferroptosis-inducing group(Erastin group), iron ferroptosis-inhibiting group(Fer-1 group), Shenfu injection group(SFI group), and Erastin+Shenfu injection group(Erastin+SFI group). 2′,7′-Dichlorodihydrofluorescein diacetate(DCFH-DA) reactive oxygen species(ROS) fluorescent probe was used to detect the ROS release level; Immunofluorescence was used to observe intracellular GPX4, DMT1 expression.
Results :
Compared with the Sham group, rats in the HIBD group showed significant neuronal cell damage in brain tissue, decreased GPX4 expression(P<0.01), increased DMT1 expression(P<0.01), decreased GSH and SOD levels(P<0.01), and increased LDH, MDA and tissue iron levels(P<0.05,P<0.05,P<0.01). In contrast, after the intervention of SFI, GPX4 expression was elevated(P<0.01), DMT1 expression decreased(P<0.01), GSH and SOD levels were elevated(P<0.01), and LDH, MDA, and tissue iron levels decreased(P<0.05,P<0.05,P<0.01). The cells experiments showed that compared with the Ctrl group, the OGD group had a significantly higher ROS content and a decrease in the expression of GPX4 fluorescence intensity, and an increase in the fluorescence intensity of DMT1(P<0.01), compared with the OGD group, the ROS content was reduced in the SFI group, while the expression of GPX4 was elevated and the expression of DMT1 was reduced(P<0.01).
Conclusion
Hippocampal and cortical regions are severely damaged after HIBD in neonatal rats, and their brain tissues show decreased expression of GPX4 and increased expression of DMT1. The above suggests that ferroptosis is involved in HIBD brain injury in neonatal rats. In contrast, Shenfu injection has a protective effect on HIBD experimental animal model and BV2 cell injury model by reducing iron aggregation and ROS production.


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