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.Full spectrum analysis of chemical constituents of Sargassum fusiforme and its in vitro anti-neuroinflammatory activity
Hong PENG ; Pinzhe HUANG ; Yonggui SONG ; Huanhua XU ; Mingyue ZHOU ; Genhua ZHU ; Ming YANG ; Zhifu AI ; Dan SU
China Pharmacy 2022;33(7):800-807
OBJECTIVE To study the composition of chemical constituents of Sargassum fusiforme and its in vitro anti- neuroinflammatory activity ,and to provide reference for its development and utilization and the study of pharmacodynamic substances. METHODS UHPLC-QTOF-MS/MS analysis method and GC-MS/MS method were used to analyze the chemical constituents of S. fusiforme . The lipopolysaccharide (1 μg/mL)was adopted to establish the inflammatory model of neuromicroglia BV2. Using paroxetine (5 μg/mL)as positive control ,CCK-8 assay was used to detect the effects of the extracts of S. fusiforme (20,40,60,80,100 μg/mL)on the activity and morphology of neuromicroglia BV 2. The effects of the extracts of S. fusiforme (40,60,80 μg/mL)on the contents of tumor necrosis factor α(TNF-α)and interleukin- 6(IL-6)in cell supernatant were detected by ELISA. RESULTS A total of 103 non-volatile constituents were identified by UHPLC-QTOF-MS/MS ,and 60 volatile constituents were obtained by GC-MS/MS. The extracts of S. fusiforme (40,60,80 μ g/mL) could significantly reduce the abnormally increased activation of neuromicroglia BV 2 and the contents of TNF-α and IL-6 due to lipopolysaccharide (P<0.05 or P<0.01). CONCLUSIONS The study establish the full spectrum of chemical constituents of S. fusiforme ,and it is confirmed that fusiforme has certain in vitro anti-neuroinflammatory activity.
3.Validation of the Thyrotoxicosis-associated Insomnia Model Induced by Thyroxine through Sympathetic Stimulation: Face, Construct and Predictive Perspectives
Zhifu AI ; Hongwei HE ; Tingting WANG ; Liling CHEN ; Chunhua HUANG ; Changlian CHEN ; Pengfei XU ; Genhua ZHU ; Ming YANG ; Yonggui SONG ; Dan SU
Experimental Neurobiology 2021;30(6):387-400
Insomnia has become a common central nervous system disease. At present, the pathogenesis of insomnia is not clear. Animal models can help us understand the pathogenesis of the disease and can be used in transformational medicine. Therefore, it is very necessary to establish an appropriate model of insomnia. Clinical data show that insomnia patients with high levels of thyroxine and often accompanied by cardiovascular problems, a common mechanism underlying all of these physiological disruptions is the sympathetic nervous system. Combined with the characteristics of chronic onset of clinical insomnia, an insomnia model induced by long-term intraperitoneal injection of thyroid hormone has been created in our laboratory. In this paper, the insomnia-like state of the model was evaluated based on three validity criteria. Face validity has been demonstrated in metabolism, the Morris water maze, electrocardiogram (ECG) and electroencephalogram (EEG). Structure validity has been proved by the results of targeted metabolomics. After treatment with diazepam, a commonly used clinical anti-insomnia drug, the above physiological and pathological disorders were reversed. The results of comprehensive analysis show that the established thyrotoxicosis-associated insomnia model meets the validity requirement to establish an appropriate animal model of insomnia. The model presented in this article might help to study pathogenetic mechanisms of clinical insomnia, as well as to test promising methods of insomnia treatment.
4.Chemical constituents from the rhizomas of Smilax glauco-china
Min LIU ; Xiaohua ZHENG ; Genhua ZHU ; Huilian HUANG ; Jicheng SHU ; Feng SHAO ; Ronghua LIU
Chinese Traditional Patent Medicine 2017;39(3):540-543
AIM To study the chemical constituents from the rhizomas of Smilax glauco-china Warb.METHODS The n-butanol fraction of ethanol extract of S.glauco-china was isolated and purified by silica,Sephadex LH-20 and semi-preparative column,then the structures of obtained compounds were identified by physicochemical properties and spectral data.RESULTS Ten compounds were isolated and identified as phenethanol-β-D-gentiobioside (1),2-phenylethyl-O-β-D-xylopyranosyl-(1 →6)-β-D-glucopyranoside (2),phenylethyl D-rutinoside (3),phenylethyl β-D-glucoside (4),hydrangeifolin Ⅰ (5),icariside D1 (6),calophymembranside B (7),2-hydroxyphenol-1-O-β-D-glucopyranosyl-(6 → 1)-α-L-rhamnopyranoside (8),β-sitosterol (9),daucosterol (10).CONCLUSION All the compounds are isolated from this plant for the first time.
5.Study on the cisplatin-sensitization effect of tripterygium glycosides on cisplatin-resistant human epithelial ovarian cancer cells (SKOV3/DDP) in vitro
Wencheng LIU ; Buzhen TAN ; Xinlu ZHAN ; Hui HU ; Genhua HUANG ; Qing CAO
Chinese Journal of Clinical Pharmacology and Therapeutics 2017;22(12):1364-1370
AIM:To study the cisplatin-sensitization effect and mechanism of tripterygium glycosides on cisplatin-resistant human epithelial ovarian cancer cells (SKOV3/DDP).METHODS:The SKOV3/DDP cells in exponential phase of growth were randomly divided into eight groups:blank control group,10 μg/mL DDP group,50 μg/mL GTW group,800 μg/mL GTW group,3 200 μg/mL GTW group,10 μg/mL DDP + 50 μg/mL GTW group,10 μg/mL DDP + 800 μg/mL GTW group and 10 μg/mL DDP + 3 200 μg/mL GTW group.Cell counting kit 8 and flow cytometry and western blot were used to detect the growth inhibiting rate and apoptosis rate and relative expression of GST-π,MDR1,STAT3,p-STAT3 of SKOV3/DDP cells of every group.RESULTS:DDP and GTW produce an additive effect when used concurrently in inhibiting growth of SKOV3/DDP cells.800 μg/mL GTW or 3 200 μg/mL GTW combined with 10 μg/mL DDP can significantly induce apoptosis of SKOV3/DDP cells and downregulate the expression of p-STAT3 (P < 0.05).CONCLUSION:GTW can significantly enhance sensitivity of SKOV3/DDP cells to DDP.The underlying mechanism may be related with down-regulating the expression of p-STAT3 in STAT3 pathway.
6.Respiratory System Infection in Heavy Type Hepatitis:Etiology Analysis of Phlegm and Prevention Measure
Genhua DONG ; Lijuan FAN ; Xiaoying HUANG
Chinese Journal of Nosocomiology 2006;0(07):-
OBJECTIVE To investigate the phlegm etiology and the prevention measures of respiratory tract infection in 89 cases with heavy type hepatitis in hospital.METHODS From Jan 2004 to May 2007,the phlegm from respiratory tract hospital infection patients were studied retrospectively.RESULTS The main respiratory tract pathogens were the G-bacillui(44.5%),G+ cocci(23.2%),Candida albicans(16.8%),Aspergillus(9.0%) and yeast-like fungi(4.5%).CONCLUSIONS The investigation of the etiology suggests that the prevention of respiratory tract infection be the main measure to decrease hospital infection in heavy type hepatitis patients.

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