1.Mechanism of Wenyang Jieyu Prescription in Regulating Activation of Mouse Hippocampal Microglia Based on JAK2/STAT3 Signaling Pathway
Ying WANG ; Zihan GONG ; Wenqing LIANG ; Jingwen YANG ; Guangxin YUE
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(8):88-96
ObjectiveTo explore the mechanism of the Wenyang Jieyu prescription in regulating depression-like behavior in mice after maternal-infant separation combined with secondary stress. MethodsAfter birth, the rats were randomly divided into blank (NC) group, maternal-infant separation (MS) group, restraint stress (RS) group, maternal-infant separation combined with restraint stress (MRS) group, Wenyang group, Jieyu group, Wenyang Jieyu (XSF) group, and minocycline group. Maternal-infant separation was performed on day 5 (PD5), followed by weaning at PD21 and prophylactic administration. The dose of Wenyang group, Xiaoyao group, XSF group and minocycline group were 5.85, 12.03, 16.71 g·kg-1 and 50 mg·kg-1, respectively. Restraint stress was applied on PD90. The model was evaluated using glucose, social interaction, open field, and O-maze behavior tests, as well as high-performance liquid chromatography to measure serotonin, dopamine, and other neurotransmitters. The expression level of ionized calcium-binding adaptor molecule-1 (Iba-1) protein, a marker of hippocampal microglia, was detected by immunohistochemistry. Protein expression levels of Janus kinase 2 (JAK2) and signal transducer and activator of transcription 3 (STAT3) in the hippocampus were analyzed by an automatic protein expression analysis system. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to detect mRNA expression levels of M1 markers, JAK2/STAT3 pathway-related genes, and cytokines in hippocampal microglia in each group. ResultsCompared with the NC group, the MRS group exhibited depression-like behavior, with significantly decreased levels of neurotransmitters in the hippocampus (P<0.05, P<0.01), increased expression of Iba-1 (P<0.01), and elevated protein levels of JAK2 and STAT3 (P<0.05). The mRNA expression levels of CD68, CD11b, IL-1β, JAK2, and STAT3 were significantly increased (P<0.01), while IL-10 mRNA expression was significantly decreased (P<0.01). Compared with the MRS group, the XSF and minocycline groups showed some improvement in depression-like behavior. In these groups, the hippocampal neurotransmitter content was significantly increased (P<0.05, P<0.01), and Iba-1 expression was significantly decreased (P<0.01). The protein levels of JAK2 and STAT3 in the XSF group showed a downward trend. The mRNA expression levels of CD68, CD11b, JAK2, STAT3, and IL-1β in the hippocampus were significantly decreased in the XSF and minocycline groups (P<0.05, P<0.01), while IL-10 mRNA expression was significantly increased (P<0.05, P<0.01). ConclusionWenyang Jieyu prescription can regulate depression-like behavior in maternal-infant separation mice combined with secondary stress by inhibiting the polarization of hippocampal microglia to the M1 phenotype. The regulation of hippocampal microglia polarization by Wenyang Jieyu prescription may be associated with the JAK2/STAT3 pathway.
2.Textual Research and Clinical Application Analysis of Classic Formula Fangji Fulingtang
Xiaoyang TIAN ; Lyuyuan LIANG ; Mengting ZHAO ; Jialei CAO ; Lan LIU ; Keke LIU ; Bingqi WEI ; Yihan LI ; Jing TANG ; Yujie CHANG ; Jingwen LI ; Bingxiang MA ; Weili DANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(11):270-277
The classic formula Fangji Fulingtang is from ZHANG Zhongjing's Synopsis of the Golden Chamber in the Eastern Han dynasty. It is composed of Stephaniae Tetrandrae Radix, Astragali Radix, Cinnamomi Ramulus, Poria, and Glycyrrhizae Radix et Rhizoma, with the effects of reinforcing Qi and invigorating spleen, warming Yang and promoting urination. By a review of ancient medical books, this paper summarizes the composition, original plants, processing, dosage, decocting methods, indications and other key information of Fangji Fulingtang, aiming to provide a literature basis for the research, development, and clinical application of preparations based on this formula. Synonyms of Fangji Fulingtang exist in ancient medical books, while the formula composition in the Synopsis of the Golden Chamber is more widespread and far-reaching. In this formula, Stephaniae Tetrandrae Radix, Astragali Radix, Cinnamomi Ramulus, Poria, and Glycyrrhizae Radix et Rhizoma are the dried root of Stephania tetrandra, the dried root of Astragalus embranaceus var. mongholicus, the dried shoot of Cinnamomum cassia, the dried sclerotium of Poria cocos, and the dried root and rhizome of Glycyrrhiza uralensis, respectively. Fangji Fulingtang is mainly produced into powder, with the dosage and decocting method used in the past dynasties basically following the original formula. Each bag is composed of Stephaniae Tetrandrae Radix 13.80 g, Astragali Radix 13.80 g, Cinnamomi Ramulus 13.80 g, Poria 27.60 g, and Glycyrrhizae Radix et Rhizoma 9.20 g. The raw materials are purified, decocted in water from 1 200 mL to 400 mL, and the decoction should be taken warm, 3 times a day. Fangji Fulingtang was originally designed for treating skin edema, and then it was used to treat impediment in the Qing dynasty. In modern times, it is mostly used to treat musculoskeletal and connective tissue diseases and circulatory system diseases, demonstrating definite effects on various types of edema and heart failure. This paper clarifies the inheritance of Fangji Fulingtang and reveals its key information (attached to the end of this paper), aiming to provide a theoretical basis for the development of preparations based on this formula.
3.Textual Research and Clinical Application Analysis of Classic Formula Fangji Fulingtang
Xiaoyang TIAN ; Lyuyuan LIANG ; Mengting ZHAO ; Jialei CAO ; Lan LIU ; Keke LIU ; Bingqi WEI ; Yihan LI ; Jing TANG ; Yujie CHANG ; Jingwen LI ; Bingxiang MA ; Weili DANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(11):270-277
The classic formula Fangji Fulingtang is from ZHANG Zhongjing's Synopsis of the Golden Chamber in the Eastern Han dynasty. It is composed of Stephaniae Tetrandrae Radix, Astragali Radix, Cinnamomi Ramulus, Poria, and Glycyrrhizae Radix et Rhizoma, with the effects of reinforcing Qi and invigorating spleen, warming Yang and promoting urination. By a review of ancient medical books, this paper summarizes the composition, original plants, processing, dosage, decocting methods, indications and other key information of Fangji Fulingtang, aiming to provide a literature basis for the research, development, and clinical application of preparations based on this formula. Synonyms of Fangji Fulingtang exist in ancient medical books, while the formula composition in the Synopsis of the Golden Chamber is more widespread and far-reaching. In this formula, Stephaniae Tetrandrae Radix, Astragali Radix, Cinnamomi Ramulus, Poria, and Glycyrrhizae Radix et Rhizoma are the dried root of Stephania tetrandra, the dried root of Astragalus embranaceus var. mongholicus, the dried shoot of Cinnamomum cassia, the dried sclerotium of Poria cocos, and the dried root and rhizome of Glycyrrhiza uralensis, respectively. Fangji Fulingtang is mainly produced into powder, with the dosage and decocting method used in the past dynasties basically following the original formula. Each bag is composed of Stephaniae Tetrandrae Radix 13.80 g, Astragali Radix 13.80 g, Cinnamomi Ramulus 13.80 g, Poria 27.60 g, and Glycyrrhizae Radix et Rhizoma 9.20 g. The raw materials are purified, decocted in water from 1 200 mL to 400 mL, and the decoction should be taken warm, 3 times a day. Fangji Fulingtang was originally designed for treating skin edema, and then it was used to treat impediment in the Qing dynasty. In modern times, it is mostly used to treat musculoskeletal and connective tissue diseases and circulatory system diseases, demonstrating definite effects on various types of edema and heart failure. This paper clarifies the inheritance of Fangji Fulingtang and reveals its key information (attached to the end of this paper), aiming to provide a theoretical basis for the development of preparations based on this formula.
4.Key Information Research and Ancient and Modern Application Analysis of Classic Prescription Houpo Sanwutang
Wenli SHI ; Qing TANG ; Huimin CHEN ; Jialei CAO ; Bingqi WEI ; Lan LIU ; Keke LIU ; Yun ZHANG ; Yujie CHANG ; Yihan LI ; Jingwen LI ; Bingxiang MA ; Lvyuan LIANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(20):214-221
Houpo Sanwutang, included in the Catalogue of Ancient Classical Prescriptions (Second Batch), was first recorded in the Synopsis of Golden Chamber written by ZHANG Zhongjing from the Eastern Han dynasty and was modified by successive generations of medical experts. A total of 37 pieces of effective data involving 37 ancient Chinese medical books were retrieved from different databases. Through literature mining, statistical analysis, and data processing, combined with modern articles, this study employed bibliometrics to investigate the historical origin, composition, decoction methods, clinical application, and other key information. The results showed that the medicinal origin of Houpo Sanwutang was clearly documented in classic books. Based on the conversion of the measurements from the Han Dynasty, it is recommended that 110.4 g Magnolia Officinalis Cortex, 55.2 g Rhei Radix et Rhizoma, and 72 g Aurantii Fructus Immaturus should be taken. Magnolia Officinalis Cortex and Aurantii Fructus Immaturus should be decocted with 2 400 mL water first, and 1 000 mL should be taken from the decocted liquid. Following this, Rhei Radix et Rhizoma should be added for further decoction, and then 600 mL should be taken from the decocted liquid. A single dose of administration is 200 mL, and the medication can be stopped when patients restore smooth bowel movement. Houpo Sanwutang has the effect of moving Qi, relieving stuffiness and fullness, removing food stagnation, and regulating bowels. It can be used in treating abdominal distending pain, guarding, constipation, and other diseases with the pathogenesis of stagnated heat and stagnated Qi in the stomach. The above results provide reference for the future development and research of Houpo Sanwutang.
5.Syndromes and Mechanisms of Depression Induced by Second Hit in Mice
Zihan GONG ; Ying WANG ; Jingwen YANG ; Wenqing LIANG ; Danhua MENG ; Kaijie SHE ; Yuan LIANG ; Guangxin YUE
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(6):29-38
ObjectiveTo explore the syndromes and mechanisms of depression induced by maternal separation (MS) combined with chronic restraint stress (RS) in mice. MethodOn postnatal day 0 (PD0), the offspring mice were randomized into a blank group (NC) and a modeling group. The mouse model of depression was established by MS+RS for 21 days. After removal of female mice on PD21, the modeled mice were randomized into model, Wenyang, Jieyu, Wenyang Jieyu, and fluoxetine groups, with 15 mice in each group. The sucrose preference, tail suspension, and open field tests were carried out to evaluate the anxiety and depression-like behavior in mice. Enzyme-linked immunosorbent assay was used to measure the adrenocorticotrophic hormone (ACTH) and corticosterone (CORT) levels in mouse plasma. High performance liquid chromatography-electrochemical detector was used to determine the content of monoamine neurotransmitters in the hippocampus. Real-time fluorescence quantitative polymerase chain reaction was employed to determine the mRNA levels of genes in the 5-hydroxytryptamine (5-HT) system, hypothalamic-pituitary-adrenal (HPA) axis, and brain-derived neurotrophic factor (BDNF) signaling pathway in the hippocampus. Immunohistochemistry was employed to determine the expression levels of proteins in the 5-HT system and HPA axis in the hippocampus. The Simple Western system was used to determine the protein levels of BDNF and tyrosine kinase receptor B (TrkB) in the hippocampus. ResultCompared with the NC group, the model group exhibited depression-like behavior, which was significantly relieved by Wenyang Jieyu prescription and fluoxetine. Compared with the NC group, the model group showed elevated levels of CORT and ACTH in the plasma (P<0.01), which, however, were lowered by Wenyang Jieyu prescription and fluoxetine (P<0.05, P<0.01). Compared with the NC group, the model group showed inhibited expression of neurotransmitters in the hippocampus (P<0.05, P<0.01), while Wenyang Jieyu prescription and fluoxetine restored the expression of neurotransmitters (P<0.05, P<0.01). Compared with NC group, the model group showed inhibition of the 5-HTergic nerve and abnormal activation of the HPA axis, and Wenyang Jieyu prescription and fluoxetine regulated the abnormal state of the 5-HTergic nerve and HPA axis. Compared with NC group, the modeling down-regulated the mRNA and protein levels of BDNF and TrkB in the hippocampus (P<0.05, P<0.01), which, however, were recovered in Wenyang, Jieyu, Wenyang Jieyu, and fluoxetine groups (P<0.05, P<0.01). ConclusionThe mouse model of depression induced by MS+RS may present the syndrome of Yang deficiency and liver depression. Wenyang Jieyu prescription may increase the content of hippocampal neurotransmitters by regulating the 5-HT system and the BDNF signaling pathway mediated by the HPA axis, thereby alleviating depression-like behavior in mice.
6.Effect of Shortening PET/CT Acquisition Time on the Diagnosis of Parkinson's Disease
Jingwen LI ; Ruiyue ZHAO ; Yimin FU ; Lifu XU ; Sihao LIANG ; Xinlu WANG
Chinese Journal of Medical Imaging 2024;32(3):240-244
Purpose To investigate the effect of shortening the acquisition time of 18F-D6-AV133 PET/CT on image quality and diagnostic efficacy in Parkinson's disease.Materials and Methods A total of 51 participants(27 of Parkinson's disease,24 of healthy-controls)from the First Affiliated Hospital of Guangzhou Medical University from October 2021 to June 2022 were retrospectively selected.Images were obtained after the injection of the tracer 18F-D6-AV133(371.04±16.30)MBq for 60 min,with collection times of 10 min.Four sets of images were reconstructed using CT attenuation correction with acquisition times of 3,5,7 and 10 min.Semi-quantitative analysis was performed on the PET images,calculating the striatum-to-occipital lobe standardized uptake value ratio(SUVR).Two physicians independently conducted qualitative evaluations for each image group.The differences of SUVR and visual score results among four sets of images were performed.The optimal critical value of SUVR was obtained by analyzing the receiver operating characteristic curve of the subjects.Results The visual analysis of image quality had a strong consistency between the two doctors(ICC=0.853,P<0.001).The images with acquisition time of 5 min could reach the common quality level in clinical work,accounting for 78.4%(40/51).The semi-quantitative results of image quality showed that there was no significant difference between SUVR and diagnostic efficiency obtained by acquisition time 5 min and 10 min(Z=1.821,P=0.069),and the best critical value of the two groups was equal to 3.Therefore,properly shortening the acquisition time had no effect on the diagnosis of Parkinson's disease.Conclusion With the development and advancement of technical equipment,PET image quality is gradually improving and the scanning time is also gradually shortening.The acquisition time for 18F-D6-AV133 can be shortened from the conventional 10 min to 5 min.
7.Comprehensive Comparative Analysis of Sepsis Animal Models and Characteristics of Clinical Syndromes in Integrated Traditional Chinese and Western Medicine
Shan LI ; Chao LI ; Yiwen WU ; Jingwen XUE ; Qingqing CHEN ; Tengxiao LIANG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(20):212-218
This study, anchored in the traditional Chinese medicine (TCM) syndrome differentiation and treatment principles alongside the clinical characteristics of sepsis in Western medicine, extensively gathers and meticulously dissects the latest research findings on sepsis animal models from both Chinese and international sources. Adhering strictly to TCM syndrome diagnostic criteria for sepsis, the study conducts a thorough evaluation of various animal models across multiple dimensions, including clinical manifestations, pathological changes, and biomarker expressions, so as to reflect the degree of resemblance these models have to human sepsis TCM syndromes. The results reveal that the colon stent implantation model exhibits a higher degree of congruence with both TCM and Western medicine standards, particularly aligning with the diagnosis of the "Fu-Qi obstruction syndrome". Conversely, the extraperitoneal sepsis model shows a higher degree of congruence with TCM, fitting more closely with the diagnosis of "acute deficiency syndrome" and emphasizing the core pathogenesis of Qi deficiency in sepsis. These findings not only augment the diversity of sepsis animal models but also highlight the necessity and potential of integrated TCM and western medicine research. Current sepsis animal models predominantly focus on western pathophysiological mechanisms, with limited direct incorporation of TCM syndrome differentiation elements. This underscores the need, in future study designs, to actively explore integrating TCM syndrome classification and intervention principles into model development. This could be achieved by manipulating model-inducing factors and observing more TCM-specific symptoms and signs among other strategies, so as to establish sepsis models that more closely resemble clinical reality and incorporate both TCM and western medical perspectives.
8.MiR-6838-5p overexpression inhibits proliferation of breast cancer MCF-7 cells by downregulating DDR1 expression
Liangjun XUE ; Qiuyu TAN ; Jingwen XU ; Lu FENG ; Wenjin LI ; Liang YAN ; Yulei LI
Journal of Southern Medical University 2024;44(9):1677-1684
Objective To explore the regulatory effect of miR-6838-5p on DDR1 gene expression and proliferation of breast cancer cells.Methods The expression levels of miR-6838-5p in normal breast epithelial cells and breast cancer cells were detected using qRT-PCR,and the potential target genes of miR-6838-5p was predicted using TargetscanV 8.0.Double luciferase reporter gene experiment was performed to verify the binding between miR-6838-5p and DDR1.Breast cancer MCF-7 cells were transfected via liposome,miR-6838-5p mimic,miR-6838-5p inhibitor,DDR1 siRNA,DDR1-overexpresisng vector,or both miR-6838-5p mimic and DDR1-overexpressing vector,and the changes in cell proliferation were examined with CCK-8 and EdU assays;Western blotting was used to detect the expression of DDR1.The mediating role of DDR1 in miR-6838-5p overexpression-induced inhibition of MCF-7 cell proliferation was verified in a nude mouse model bearing MCF-7 cell xenografts.Results The expression of miR-6838-5p was significantly lower in breast cancer cells than in normal breast epithelial cells.In MCF-7 cells,miR-6838-5p overexpression induced significant inhibition of cell proliferation.Dual luciferase reporter gene experiment demonstrated a binding relationship between miR-6838-5p and DDR1(P<0.01).Western blotting showed that miR-6838-5p overexpression significantly lowered DDR1 expression in MCF-7 cells,and DDR1 overexpression promoted proliferation of the cells;co-transfection of the cells with DDR1-overexpressing vector significantly attenuated the inhibitory effect of miR-6838-5p mimic on cell proliferation.In the tumor-bearing nude mice,the xenografts overexpressing miR-6838-5p showed a significantly smaller volum with obviously the expression of DDR1.Conclusion Overexpression of miR-6838-5p inhibits breast cancer cell proliferation by regulating DDR1 expression.
9.Reduced expression of semaphorin 3A in osteoclasts causes lymphatic expansion in a Gorham-Stout disease(GSD)mouse model
ZHANG DONGFANG ; XU HAO ; QIN CHI ; CAI KANGMING ; ZHANG JING ; XIA XINQIU ; BI JINGWEN ; ZHANG LI ; XING LIANPING ; LIANG QIANQIAN ; WANG WENSHENG
Journal of Zhejiang University. Science. B 2024;25(1):38-50,中插1-中插8
Gorham-Stout disease(GSD)is a sporadic chronic disease characterized by progressive bone dissolution,absorption,and disappearance along with lymphatic vessel infiltration in bone-marrow cavities.Although the osteolytic mechanism of GSD has been widely studied,the cause of lymphatic hyperplasia in GSD is rarely investigated.In this study,by comparing the RNA expression profile of osteoclasts(OCs)with that of OC precursors(OCPs)by RNA sequencing,we identified a new factor,semaphorin 3A(Sema3A),which is an osteoprotective factor involved in the lymphatic expansion of GSD.Compared to OCPs,OCs enhanced the growth,migration,and tube formation of lymphatic endothelial cells(LECs),in which the expression of Sema3A is low compared to that in OCPs.In the presence of recombinant Sema3A,the growth,migration,and tube formation of LECs were inhibited,further confirming the inhibitory effect of Sema3A on LECs in vitro.Using an LEC-induced GSD mouse model,the effect of Sema3A was examined by injecting lentivirus-expressing Sema3A into the tibiae in vivo.We found that the overexpression of Sema3A in tibiae suppressed the expansion of LECs and alleviated bone loss,whereas the injection of lentivirus expressing Sema3A short hairpin RNA(shRNA)into the tibiae caused GSD-like phenotypes.Histological staining further demonstrated that OCs decreased and osteocalcin increased after Sema3A lentiviral treatment,compared with the control.Based on the above results,we propose that reduced Sema3A in OCs is one of the mechanisms contributing to the pathogeneses of GSD and that expressing Sema3A represents a new approach for the treatment of GSD.
10.Application of SAT-TB in the evaluation of curative effect of initial treatment of smear-positive pulmonary tuberculosis patients
Jingwen LAI ; Kehao PENG ; Liuyue XU ; Xiaoyu LAI ; Jingzhong LI ; Meiling YU ; Hongdi LIANG ; Xunxun CHEN
International Journal of Laboratory Medicine 2024;45(21):2577-2581,2587
Objective To evaluate the application value of simultaneous amplification and testing for Myco-bacterium tuberculosis(SAT-TB)in evaluating the curative effect of initial treatment of smear-positive pul-monary tuberculosis patients.Methods A total of 62 newly treated smear positive pulmonary tuberculosis pa-tients from June 2022 to June 2023 in Guangzhou Panyu District Chronic Disease Control Station were selected as the study objects,and the curative effect was evaluated by liquid-based sandwich cup method,Roche culture method and SAT-TB detection method.All patients received the standard anti-tuberculosis treatment regimen of 2HRZE/4HR standard regimen,and sputum samples were detected by liquid-based sandwich cup method,Roche culture method,and SAT-TB detection method at the 2nd,4th,and 8th week of treatment,respectively.Results Among 62 patients,54 cases were positive and 8 cases were negative using Roche culture method,47 cases were positive and 15 cases were negative using SAT-TB detection method.60 cases were positive and 2 cases were negative by Mycobacterium tuberculosis(MTB)-DNA test.The positive rates of the three methods were 87.10%(54/62),75.81%(47/62)and 96.77%(60/62),respectively.Taking Roche culture method re-sults as the standard,the sensitivity of SAT-TB detection method and MTB-DNA was 97.87%(46/47)and 90.00%(54/60),and the specificity was 46.67%(7/15)and 100.00%(2/2),respectively.There were signifi-cant differences between Roche culture method and SAT-TB detection method and MTB-DNA test(x2=20.070,P<0.05,x2=13.985,P<0.05),the difference between the results of SAT-TB detection method and MTB-DNA test was also statistically significant(x2=8.365,P<0.05).The negative conversion rates of MTB in sputum samples were 69.35%(43/62),29.03%(18/62)and 41.94%(26/62)at the 2nd,4 th,and 8 th weeks,respectively.77.42%(48/62),59.68%(37/62),58.06%(36/62),82.26%(51/62),79.03%(49/62),75.81%(47/62).There were significant differences in the negative conversion rates of MTB in sputum sam-ples between SAT-TB and liquid-based sandwich cup method at the 2nd and 4th weeks(x2=8.365,P<0.05,x2=4.465,P<0.05),while there was no significant difference between the results of Roche culture method at the 2nd,4th and 8th weeks(x2=1.726,P>0.05,x2=0.000,P>0.05,x2=0.046,P>0.05).Conclusion The use of SAT-TB detection method in clinical practice to evaluate the efficacy of smear positive pulmonary tuberculo-sis patients could accurately and quickly assess the negative conversion rate and treatment effect of patients,and provide a reliable basis for guiding clinical treatment.It could be considered as an effective auxiliary diag-nosis and evaluation method of curative effect,worthy of promotion and practical application.

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