1.Untargeted Metabolomics Reveals Mechanism of Modified Sinisan in Ameliorating Anxiety-like Behaviors Induced by Chronic Restraint Stress in Mice
Jie ZHAO ; Zhengyu FANG ; He XIAO ; Na GUO ; Hongwei WU ; Hongjun YANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(21):70-79
ObjectiveTo elucidate the potential mechanism of modified Sinisan (MSNS) in alleviating anxiety-like behaviors induced by chronic restraint stress (CRS) in mice at the metabolic level based on serum untargeted metabolomics and identify key metabolites and metabolic pathways regulated by MSNS. MethodsSeventy-two male C57BL/6 mice were randomly assigned into six groups: control, model, high-dose (2.4 g·kg-1) MSNS, medium-dose (1.2 g·kg-1) MSNS, low-dose (0.6 g·kg-1) MSNS, and positive control (fluoxetine, 2.6 mg·kg-1). Except the control group, the other groups were subjected to CRS for the modeling of anxiety. Mice were administrated with corresponding agents by gavage 2 h before daily restraint for 14 days. Anxiety-like behaviors were evaluated by the open field test (OFT), elevated plus maze (EPM) test, and light/dark box (LDB) test. Serum levels of corticotropin-releasing hormone (CRH), adrenocorticotrophic hormone (ACTH), and corticosterone (CORT) were measured via ELISA to assess stress levels. Ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was employed to detect 9 metabolites in the brain tissue and serum metabolites. Orthogonal partial least squares-discriminant analysis (OPLS-DA) was adopted to identify differential metabolites (VIP>1.0, P<0.05). MetaboAnalyst 5.0 was used for metabolic pathway enrichment analysis of the differential metabolites. ResultsCompared with the control group, the model group showed reductions in the central activity time and central distance in the OFT (P<0.05), the proportions of open-arm residence time and open-arm residence times in the EPM test (P<0.01), and the proportions of open box activity time and open box activity distance in the LDB test (P<0.05), which were increased in the medium- and high-dose MSNS groups compared with the model group (P<0.05). Compared with the control group, the model group showed elevated levels of CRH, ACTH, and CORT in the serum (P<0.01), and the elevations were diminished in the medium- and high-dose MSNS groups (P<0.05). UPLC-MS results indicated that compared with the control group, the model group presented declined DA, GABA, 5-HIAA, 5-HT, and 5-HT/Trp levels (P<0.05, P<0.01) and raised Glu, NE, Kyn, and Kyn/Trp levels (P<0.05). Compared with the model group, high-dose MSNS increased the GABA, 5-HIAA, and 5-HT/Trp levels (P<0.05) and lowered the Glu and Kyn/Trp levels (P<0.05). Untargeted metabolomics identified that 16 CRS-induced metabolic disturbances were reversed by MSNS. KEGG pathway analysis indicated that MSNS primarily modulated eight core pathways including alanine/aspartate/glutamate metabolism, butyrate metabolism, arginine-proline metabolism, TCA cycle, unsaturated fatty acid biosynthesis, and tryptophan metabolism. The mechanisms involved multidimensional biological processes, including neurotransmitter homeostasis regulation, TCA cycle energy metabolism optimization, and inflammatory response suppression. ConclusionMSNS alleviates CRS-induced anxiety-like behaviors in mice by mitigating hypothalamic-pituitary-adrenal axis hyperactivity, improving hippocampal neurotransmitter and tryptophan metabolic pathways, and regulating alanine/aspartate/glutamate metabolism, butyrate metabolism, arginine-proline metabolism, and TCA cycle.
2.Untargeted Metabolomics Reveals Mechanism of Modified Sinisan in Ameliorating Anxiety-like Behaviors Induced by Chronic Restraint Stress in Mice
Jie ZHAO ; Zhengyu FANG ; He XIAO ; Na GUO ; Hongwei WU ; Hongjun YANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(21):70-79
ObjectiveTo elucidate the potential mechanism of modified Sinisan (MSNS) in alleviating anxiety-like behaviors induced by chronic restraint stress (CRS) in mice at the metabolic level based on serum untargeted metabolomics and identify key metabolites and metabolic pathways regulated by MSNS. MethodsSeventy-two male C57BL/6 mice were randomly assigned into six groups: control, model, high-dose (2.4 g·kg-1) MSNS, medium-dose (1.2 g·kg-1) MSNS, low-dose (0.6 g·kg-1) MSNS, and positive control (fluoxetine, 2.6 mg·kg-1). Except the control group, the other groups were subjected to CRS for the modeling of anxiety. Mice were administrated with corresponding agents by gavage 2 h before daily restraint for 14 days. Anxiety-like behaviors were evaluated by the open field test (OFT), elevated plus maze (EPM) test, and light/dark box (LDB) test. Serum levels of corticotropin-releasing hormone (CRH), adrenocorticotrophic hormone (ACTH), and corticosterone (CORT) were measured via ELISA to assess stress levels. Ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was employed to detect 9 metabolites in the brain tissue and serum metabolites. Orthogonal partial least squares-discriminant analysis (OPLS-DA) was adopted to identify differential metabolites (VIP>1.0, P<0.05). MetaboAnalyst 5.0 was used for metabolic pathway enrichment analysis of the differential metabolites. ResultsCompared with the control group, the model group showed reductions in the central activity time and central distance in the OFT (P<0.05), the proportions of open-arm residence time and open-arm residence times in the EPM test (P<0.01), and the proportions of open box activity time and open box activity distance in the LDB test (P<0.05), which were increased in the medium- and high-dose MSNS groups compared with the model group (P<0.05). Compared with the control group, the model group showed elevated levels of CRH, ACTH, and CORT in the serum (P<0.01), and the elevations were diminished in the medium- and high-dose MSNS groups (P<0.05). UPLC-MS results indicated that compared with the control group, the model group presented declined DA, GABA, 5-HIAA, 5-HT, and 5-HT/Trp levels (P<0.05, P<0.01) and raised Glu, NE, Kyn, and Kyn/Trp levels (P<0.05). Compared with the model group, high-dose MSNS increased the GABA, 5-HIAA, and 5-HT/Trp levels (P<0.05) and lowered the Glu and Kyn/Trp levels (P<0.05). Untargeted metabolomics identified that 16 CRS-induced metabolic disturbances were reversed by MSNS. KEGG pathway analysis indicated that MSNS primarily modulated eight core pathways including alanine/aspartate/glutamate metabolism, butyrate metabolism, arginine-proline metabolism, TCA cycle, unsaturated fatty acid biosynthesis, and tryptophan metabolism. The mechanisms involved multidimensional biological processes, including neurotransmitter homeostasis regulation, TCA cycle energy metabolism optimization, and inflammatory response suppression. ConclusionMSNS alleviates CRS-induced anxiety-like behaviors in mice by mitigating hypothalamic-pituitary-adrenal axis hyperactivity, improving hippocampal neurotransmitter and tryptophan metabolic pathways, and regulating alanine/aspartate/glutamate metabolism, butyrate metabolism, arginine-proline metabolism, and TCA cycle.
3.Cigarette Smoke Extract-TreatedMouse Airway Epithelial Cells-Derived Exosomal LncRNA MEG3 Promotes M1 Macrophage Polarization and Pyroptosis in Chronic Obstructive Pulmonary Disease by Upregulating TREM-1 via m6A Methylation
Lijing WANG ; Qiao YU ; Jian XIAO ; Qiong CHEN ; Min FANG ; Hongjun ZHAO
Immune Network 2024;24(2):e3-
Cigarette smoke extract (CSE)-treated mouse airway epithelial cells (MAECs)-derived exosomes accelerate the progression of chronic obstructive pulmonary disease (COPD) by upregulating triggering receptor expressed on myeloid cells 1 (TREM-1); however, the specific mechanism remains unclear. We aimed to explore the potential mechanisms of CSE-treated MAECs-derived exosomes on M1 macrophage polarization and pyroptosis in COPD. In vitro, exosomes were extracted from CSE-treated MAECs, followed by co-culture with macrophages.In vivo, mice exposed to cigarette smoke (CS) to induce COPD, followed by injection or/and intranasal instillation with oe-TREM-1 lentivirus. Lung function and pathological changes were evaluated. CD68 + cell number and the levels of iNOS, TNF-α, IL-1β (M1 macrophage marker), and pyroptosis-related proteins (NOD-like receptor family pyrin domain containing 3, apoptosis-associated speck-like protein containing a caspase-1 recruitment domain, caspase-1, cleaved-caspase-1, gasdermin D [GSDMD], and GSDMD-N) were examined. The expression of maternally expressed gene 3 (MEG3), spleen focus forming virus proviral integration oncogene (SPI1), methyltransferase 3 (METTL3), and TREM-1 was detected and the binding relationships among them were verified. MEG3 increased N6-methyladenosine methylation of TREM-1 by recruiting SPI1 to activate METTL3. Overexpression of TREM-1 or METTL3 negated the alleviative effects of MEG3 inhibition on M1 polarization and pyroptosis. In mice exposed to CS, EXO−CSE further aggravated lung injury, M1 polarization, and pyroptosis, which were reversed by MEG3 inhibition. TREM-1 overexpression negated the palliative effects of MEG3 inhibition on COPD mouse lung injury. Collectively, CSE-treated MAECs-derived exosomal long non-coding RNA MEG3 may expedite M1 macrophage polarization and pyroptosis in COPD via the SPI1/METTL3/TREM-1 axis.
4.Herbal Textual Research on Angelicae Pubescentis Radix and Notopterygii Rhizoma et Radix in Famous Classical Formulas
Xiao DU ; Yapeng WANG ; Jinxiu QIAN ; Hongjun YANG ; Huihui LIU ; Zhilai ZHAN
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(5):68-83
By reviewing ancient materia medica, prescription and medical books, combined with modern literature, the paper made textual research on the name, origin, producing area, quality evaluation, harvesting and processing methods of Angelicae Pubescentis Radix and Notopterygii Rhizoma et Radix, so as to provide a basis for the selection and use of these two herbs in the development of famous classical formulas. Through textual research, it can be found that Angelicae Pubescentis Radix and Notopterygii Rhizoma et Radix were mixed together in the early history of China, but the distinction was first made during the Southern and Northern dynasties, and since then there have been constant controversies, and it is not until contemporary times that they are distinguished clearly. In the past dynasties, Duhuo and Qianghuo were used as the rectification of names, some aliases and trade names were also seen. Angelica biserrata is the mainstream origin of Angelicae Pubescentis Radix in the past dynasties, and there are many plants belonging to Angelica, Heracleum and Aralia, which are also used as this medicine. However, the origin of Notopterygii Rhizoma et Radix used in the past dynasties is mostly Notopterygium incisum or N. franchetii, which is relatively uniform. The producing areas of Angelicae Pubescentis Radix and Notopterygii Rhizoma et Radix are mostly concentrated in the western and northwestern regions of China, among which Angelicae Pubescentis Radix is mainly produced in Hubei, Chongqing, Sichuan, Shaanxi and other places, and the border area between Hubei and Chongqing is the geo-authentic area. Notopterygii Rhizoma et Radix is mainly produced in Sichuan, Gansu, Qinghai, Shaanxi and others with the western and northern Sichuan and southern Gansu as the geo-authentic areas. Angelicae Pubescentis Radix and Notopterygii Rhizoma et Radix in the past dynasties were harvested in spring and autumn, especially in February and August of the lunar calendar. Angelicae Pubescentis Radix with strong main roots, few branches, firm texture and strong aroma is superior, and Notopterygii Rhizoma et Radix with strong rhizomes, tightly raised knots, purple-brown skin, tight cross-section, strong aroma and silkworm-like shape is superior. The processing methods of Angelicae Pubescentis Radix and Notopterygii Rhizoma et Radix are mostly cut after cutting the reeds, and the raw product is used as medicine. Based on the above research results, it is recommended that the roots of A. biserrata should be used for Angelicae Pubescentis Radix and the roots of N. incisum should be used for Notopterygii Rhizoma et Radix in the development of famous classical formulas, and raw products should be used in the formulas that do not specify processing requirements.
5.Analysis of wideband acoustic immittance measurements in patients with unilateral Ménière's disease.
Hui PAN ; Min XIE ; Bo LIU ; Hongjun XIAO
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2023;37(10):809-814
Objective:To investigate the characteristics of wideband acoustic immittance(WAI) measurements in patients with unilateral Ménière's disease(MD) and evaluate the clinical value of WAI in diagnosis of MD. Methods:WAI was performed in 30 patients with unilateral MD(30 ears for symptomatic and 30 ears for asymptomatic) and in 26 healthy individuals(52 ears)(control group). The WAI measurements, including the frequency first appearing two peaks in energy absorbance(EA) tympanogram, resonance frequency(RF), the peak value of absorbance(PVA), the integral area of absorbance(IAA), EA curve at peak pressure, were analyzed. Results:①The occurrence of two peaks in EA tympanogram in both the MD symptomatic and asymptomatic ear was observed in 27 ears(84.4%), and 38 ears(70.4%) in the control group, with no significant difference in the frequency of first appearing in two peaks onset between the groups(all P>0.05). ②The RF of the MD symptomatic ears was significantly lower than that of the asymptomatic ears(t=-3.544, P=0.001) and that of the control subjects(t=2.084, P=0.041); there was no difference of RF between the MD asymptomatic ears and the control group(P>0.05). ③The PVA were significantly lower in both MD symptomatic(t=4.240, P<0.01) and asymptomatic ears(t=4.202, P=0.001) than in controls. ④The IAA in MD symptomatic(t=3.295, P=0.001) and asymptomatic ears(t=3.193, P=0.003) was significantly lower than in the control group. ⑤Comparison of the EA curve at peak pressure of the three groups: the EAs of MD symptomatic ears were lower than those of the control group at the range of 1 059-2 911 Hz(all P<0.05); the EAs of MD symptomatic ears were lower than those of MD asymptomatic ears within 1 000 Hz and 1 834-2 119 Hz(all P<0.05); the EAs of MD asymptomatic ears were lower than those of the control group at the range of 515-2 748 Hz(all P<0.05). Conclusion:Symptomatic ears in unilateral MD patients show alterations in some WAI measurements compared to asymptomatic ears and/or controls, suggesting that middle ear mechanical fuction of the affected side may be modified due to the endolymphatic hydrops. The clinical significance of WAI needs to be further explored in the context of evaluating MD.
Humans
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Meniere Disease/diagnosis*
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Endolymphatic Hydrops/diagnosis*
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Ear
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Hearing Tests
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Acoustics
6.A case report of prostate mucinous adenocarcinoma
Xinyuan YU ; Xiao LI ; Hongjun ZHAO ; Bao LI ; Huimin SUN ; Quan ZHOU
Chinese Journal of Urology 2023;44(3):224-225
Mucinous adenocarcinoma of the prostate is a rare pathological type of prostate cancer. We reported one case. The patient went to see a doctor because of intermittent hematuria. He was diagnosed as prostate adenocarcinoma before operation. He underwent laparoscopic radical prostatectomy. Postoperative pathological examination showed mucinous adenocarcinoma of the prostate. The patient was followed up for six months. Imaging showed no signs of recurrence and metastasis with normal tPSA.
7.Epidemiological investigation of a case with SARS-CoV-2 infection associated with overseas countries at an international harbor
Dongliang ZHANG ; Bo YI ; Yi CHEN ; Qunxiong HU ; Feng LING ; Xiao MA ; Song LEI ; Hongjun DONG ; Hongxia NI ; Yang MAO ; Qiaofang LI ; Yaorong CHEN ; Ye LU ; Zhenyu GONG ; Jian CAI ; Zhiping CHEN ; Jun LÜ ; Guozhang XU
Journal of Preventive Medicine 2022;34(4):380-384
Objective:
To investigate the origin of infection and risk factors of a case with SARS-CoV-2 infection associated with overseas countries in the Ningbo-Zhoushan Port, Zhejiang Province, so as to provide the evidence for improving the COVID-19 control measures at ports.
Methods:
Ningbo Center for Disease Control and Prevention ( CDC ) and Beilun CDC conducted case finding and epidemiological surveys immediately after being informed. The general information, history of vaccination and the travel during the latest 14 days were collected from the positive case, and all close contacts were tracked. Saliva samples were collected for SARS-CoV-2 nucleic acid testing and whole-genome sequencing, and the sequencing results were aligned with the GISAID's EpiCoV database. The origin of infection and transmission route of the positive case was investigated.
Results:
A case was identified positive for SARS-CoV-2 nucleic acid during company M's routine screening in the Ningbo-Zhoushan Port on August 10, 2021, and was confirmed positive for SARS-CoV-2 nucleic acid by Beilun CDC and Ningbo CDC on August 11. Whole-genome sequencing showed SARS-CoV-2 B.1.617.2 ( Delta ) variant, which shared the highest homology with the virus sequence uploaded by Russia on June, 2021 ( Russia/MOW-RII-MH27356S/2021 ). The case was a bundling worker for overseas container ships, and reported communicated with foreign boatmen and contacted materials without protected interventions on the SINOKOR AKITA Container Ship between August 4 and 5, 2021. This ship anchored at Vladivostok, Russia from July 27 to 29, anchored at Ningbo Harbor on August 4, and departed on August 5. Then, 11 boatmen from this ship were tested positive for SARS-CoV-2 nucleic acid on August 8. One asymptomatic case was reported in this epidemic; 254 close contacts and 617 secondary close contacts were identified, and all were tested negative for SARS-CoV-2 nucleic acid. No new cases with SARS-CoV-2 infections were detected until August 25, 2021, and the emergency response was therefore terminated.
Conclusions
The infection was a sporadic COVID-19 epidemic associated with overseas countries, which was caused by Delta variant infection through contacts with foreign boatmen or materials by a bundling worker in Ningbo-Zhoushan Port; fortunately, no epidemic spread occurred. Intensified closed-loop management and increased frequency of SARS-CoV-2 nucleic acid test among high-risk populations, and improving the precision and rapid emergency treatment of COVID-19 epidemics are required for the containment of COVID-19 at ports.
8.Epidemiological characteristics of coronavirus disease 2019 in Ningbo
ZHANG Dongliang ; YI Bo ; CHEN Yi ; DING Keqin ; WANG Haibo ; DONG Hongjun ; XU Guozhang ; WANG Aihong ; MA Xiao ; ZHANG Yan ; FANG Ting
Journal of Preventive Medicine 2020;32(4):330-333
Objective:
To analyze the epidemiological characteristics of coronavirus disease 2019 (COVID-19) cases reported in Ningbo from January 22 to February 22, 2020, so as to provide guidance for the prevention and control of the COVID-19 epidemic.
Methods:
The confirmed cases of COVID-19 reported by Ningbo were selected from National Diseases Prevention and Control Information System to analyze the epidemiological characteristics by descriptive epidemiological method, including time, spatial and population distribution, clinical symptoms and exposure history.
Results:
A total of 157 confirmed cases of COVID-19 were reported and there was no death. The first confirmed case was reported on January 22. On the incidence curve, the peak was from January 22 to February 4, with a maximum of 15 cases in a single day. The incidence curve presented sustained human-to-human transmission. The number of the cases showed a declining trend from February 5. The confirmed cases were reported in all 10 counties(cities or districts),among which 69(43.95%)cases were reported in Haishu District. The confirmed cases were mainly 30-69 years old,accounting for 78.34%;were mainly farmers,household workers and retired people,accounting for 59.87%;and were mainly clinically mild,accounting for 87.90%.There were 51 imported cases, accounting for 32.48%. The initial confirmed case was a local case. In the early stage of the epidemic, local cases and imported cases prevalent together,which was due to the outbreak caused by the large-scale buddhist activity on January 19.This event resulted in 67 confirmed cases and 15 asymptomatic cases. There totally reported 22 clusters with 138 (87.90%)confirmed cases.
Conclusions
In the early stage of the COVID-19 epidemic in Ningbo, the imported cases and local cases prevalent at the same time. Most of the cases were female, aged from 30 to 69 years, and their occupation were farmers, household workers and retired people. With comprehensive measures taken, the epidemic of COVID-19 in Ningbo have been under control.
9.Clinical significance of magnetic resonance spectroscopy in patients with prostate specific antigenabnormal prostate
Chao SONG ; Yuliang WANG ; Xiao ZHENG ; Xueping ZHANG ; Yongjian JI ; Yang ZHOU ; Hongjun ZHAO ; Yongxiang LI ; Boli Liang2Lin QIAO
Chinese Journal of Postgraduates of Medicine 2019;42(6):543-546
Objective To evaluate the diagnostic value of magnetic resonance spectroscopy analysis in patients with prostate specific antigen (PSA) abnormal prostate disease. Methods The patients who had abnormal PSA from January 2017 to December 2018 in Weifang People′s Hospital were selected. Patients with prostate puncture indications were as research target. A total of 137 patients including 44 patients with prostate cancer and 93 patients with benign prostatic hyperplasia were diagnosed by puncture. All patients underwent magnetic resonance scan and enhancement and spectroscopy before surgery. The comparative value of magnetic resonance scan + enhancement, magnetic resonance spectroscopy and joint examination was obtained. Results Sensitivity of magnetic resonance imaging and enhancement was 77.3% (34/44), specificity was 86.0% (80/93), and accuracy was 83.2% (114/137). Sensitivity of magnetic resonance spectroscopy was 52.3% (23/44), specificity was 77.4% (72/93), and accuracy was 69.3% (95/137). The sensitivity of the combined application was 90.9% (40/44), specificity was 91.4% (85/93), and accuracy was 91.2% (125/137). Conclusions The application of magnetic resonance spectroscopy can increase the sensitivity, specificity and accuracy of magnetic resonance plain scan and intensive examination for diagnosis of prostate cancer.
10.Study on the Attitude towards the Protection of Traditional Knowledge of Chinese Medicine among Students in a University of Traditional Chinese Medicine in Shaanxi Province
Xiao TANG ; Jun LIANG ; Xinxin FENG ; Hongjun HOU ; Yuan ZHANG ; Xiaoqin WANG ; Jianan FENG ; Mingxu WANG
Chinese Medical Ethics 2018;31(6):693-699
Objective:To investigate and analyze the attitude and its influencing factors of traditional knowledge of Chinese medicine among students in a Traditional Chinese Medicine ( TCM) college in Shaanxi Province, and provide scientific evidence for improving the relevant institution and policies. Method:A stratified cluster sampling method was used to conduct a questionnaire survey of 1,162 students in a TCM college in Shaanxi Province. Rank sum test and multiple linear regression model were used to explore the factors affecting the attitude of students. Result:The score of attitudes towards TCM protection was(43. 09 ± 6. 50), while specialty, parental education, family members' attitudes towards TCM, evaluation of courses in TCM, had taken related courses or not, clinical practice or internship experience of TCM, and the score of correlative knowledge were the influencing factors of attitudes ( P<0 . 05 ) . Conclusion:Students of this school had a relatively positive attitude towards the protection of traditional knowledge of Chinese medicine. In order to cultivate the fresh troop for the development of TCM, it is suggested that the system of TCM courses and textbooks should be reformed so as to improve students' enthusiasm for protecting traditional knowledge of Chinese medicine and establishing a special institution.


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