1.In Vitro and in vivo Component Analysis of Total Phenolic Acids from Gei Herba and Its Effect on Promoting Acute Wound Healing and Inhibiting Scar Formation
Xixian KONG ; Guanghuan TIAN ; Tong WU ; Shaowei HU ; Jie ZHAO ; Fuzhu PAN ; Jingtong LIU ; Yong DENG ; Yi OUYANG ; Hongwei WU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(3):156-167
ObjectiveBased on ultra performance liquid chromatography-quadrupole-electrostatic field orbital trap high-resolution mass spectrometry(UPLC-Q-Orbitrap-MS), to identify the in vivo and in vitro chemical components of total phenolic acids in Gei Herba(TPAGH), and to clarify the pharmacological effects and potential mechanisms of the effective part in promoting acute wound healing and inhibiting scar formation. MethodsUPLC-Q-Orbitrap-MS was used to identify the chemical components of TPAGH and ingredients absorbed in vivo after topical administration. A total of 120 ICR mice were randomly divided into the model group, recombinant human epidermal growth factor(rhEGF) group(4 mg·kg-1), and low, medium, and high dose groups of TPAGH(3.5, 7, 14 mg·kg-1), with 24 mice in each group. A full-thickness skin excision model was constructed, and each administration group was coated with the drug at the wound site, and the model group was treated with an equal volume of normal saline, the treatment was continued for 30 days, during which 8 mice from each group were sacrificed on days 6, 12, and 30. The healing of the wounds in the mice was observed, and histopathological changes in the skin tissues were dynamically observed by hematoxylin-eosin(HE), Masson, and Sirius red staining, and enzyme-linked immunosorbent assay(ELISA) was used to dynamically measure the contents of interleukin-6(IL-6), tumor necrosis factor-α(TNF-α), vascular endothelial growth factor A(VEGFA), matrix metalloproteinase(MMP)-3 and MMP-9 in skin tissues. Network pharmacology was used to predict the targets related to the promotion of acute wound healing and the inhibition of scar formation by TPAGH, and molecular docking of key components and targets was performed. Gene Ontology(GO) biological process analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analysis were carried out for the related targets, so as to construct a network diagram of herbal material-compound-target-pathway-pharmacological effect-disease for further exploring its potential mechanisms. ResultsA total of 146 compounds were identified in TPAGH, including 28 phenylpropanoids, 31 tannins, 23 triterpenes, 49 flavonoids, and 15 others, and 16 prototype components were found in the serum of mice. Pharmacodynamic results showed that, compared with the model group, the TPAGH groups showed a significant increase in relative wound healing rate and relative scar inhibition rate(P<0.05), and the number of new capillaries, number of fibroblasts, number of new skin appendages, epidermal regeneration rate, collagen deposition ratio, and Ⅲ/Ⅰ collagen ratio in the tissue were significantly improved(P<0.05, 0.01), the levels of IL-6, TNF-α, MMP-3 and MMP-9 in the skin tissues were reduced to different degrees, while the level of VEGFA was increased. Network pharmacology analysis screened 10 core targets, including tumor protein 53(TP53), sarcoma receptor coactivator(SRC), protein kinase B(Akt)1, signal transducer and activator of transcription 3(STAT3), epidermal growth factor receptor(EGFR) and so on, participating in 75 signaling pathways such as advanced glycation end-products(AGE)-receptor for AGE(AGE/RAGE) signaling pathway, phosphatidylinositol 3-kinase(PI3K)/Akt signaling pathway, mitogen-activated protein kinase(MAPK) signaling pathway. Molecular docking confirmed that the key components genistein, geraniin, and casuariin had good binding ability to TP53, SRC, Akt1, STAT3 and EGFR. ConclusionThis study comprehensively reflects the chemical composition of TPAGH and the absorbed components after topical administration through UPLC-Q-Orbitrap-MS. TPAGH significantly regulates key indicators of skin healing and tissue reconstruction, thereby clarifying its role in promoting acute wound healing and inhibiting scar formation. By combining in vitro and in vivo component identification with network pharmacology, the study explores how key components may bind to targets such as TP53, Akt1 and EGFR, exerting therapeutic effects through related pathways such as immune inflammation and vascular regeneration.
2.Mechanism of Shengmai Injection Against Cerebral Ischemia Based on Proteomics
Jingtong LIU ; Shaowei HU ; Mengli CHANG ; Jing XU ; Qingqing CAI ; Xinghong LI ; Liying TANG ; Huanhuan WANG ; Hongwei WU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(9):57-67
ObjectiveTo evaluate pharmacological effects of Shengmai injection(SMI)on cerebral ischemia and study its neuroprotective mechanism. MethodsMale specific pathogen-free (SPF) Sprague-Dawley (SD) rats were randomly divided into a sham group, a model group, a low-dose SMI group(3 mL·kg-1), a middle-dose SMI group(6 mL·kg-1), a high-dose SMI group(12 mL·kg-1), and a Ginaton group(4 mL·kg-1)according to the random number table method, with 12 rats in each group. The rat model of cerebral ischemia-reperfusion(MCAO/R)was prepared via the suture method. The administration groups were intraperitoneally injected with corresponding concentrations of SMI or Ginaton injection after reperfusion, which was conducted for 3 consecutive days. The sham group and model group were administered the equivalent volume of physiological saline. The pharmacological effects of SMI on brain injury in MCAO/R rats were evaluated by neurological function scores, cerebral infarction area, hematoxylin-eosin (HE) staining, Nissl staining, terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) staining, and Western blot. The dominant link and key protein of SMI treating cerebral injury were explored using proteomic analysis. The related mechanisms of SMI were further validated using enzyme-linked immunosorbent assay (ELISA), Western blot, and chloride ion fluorescence probe with oxygen-glucose deprivation/reoxygenation(OGD/R)-treated PC12 cells and MCAO/R rats. ResultsCompared with the sham group, the model group showed significantly increased neurological function scores, cerebral infarction area, neuronal apoptosis rate, and expression levels of apoptosis related proteins (P<0.05, P<0.01)and significantly decreased density of Nissl bodies and neurons(P<0.01). Compared with the model group, the SMI groups exhibited significantly decreased neurological function scores, cerebral infarction area, neuronal apoptosis rate, and expression levels of apoptosis related proteins (P<0.05, P<0.01)and significantly increased density of Nissl bodies and neurons (P<0.05). The proteomic analysis results showed that oxidative stress and inflammatory response were important processes of SMI intervening in MCAO/R injury, and the chloride intracellular channel protein 1 (CLIC1) was one of key proteins in its action network. The levels of representative indicators of oxidative stress and inflammatory response in the MCAO/R rats of the SMI groups were significantly reduced, compared with those in the model group(P<0.05, P<0.01), and the expression levels of CLIC1 and downstream NOD-like receptor protein 3 (NLRP3) decreased (P<0.01). In addition, the experimental results based on the OGD/R PC12 cells showed that SMI significantly increased the cell survival rate(P<0.01) and significantly decreased the intracellular chloride ion concentration(P<0.05). ConclusionSMI has neuroprotective effects. Oxidative stress and inflammatory response are key processes of SMI intervening in MCAO/R injury. The potential mechanism is closely related to the regulation of CLIC1.
4.Association between vertebral artery hypoplasia and posterior circulation infarction:Progresses of imaging researches
Jingtong XIONG ; Lingjun MEI ; Jiawen LUO ; Xiaofeng QU ; Jianlin WU
Chinese Journal of Medical Imaging Technology 2024;40(7):1104-1107
Vertebral artery hypoplasia(VAH)may lead changes of hemodynamic status in the posterior circulation,promote occurrence of intracranial atherosclerotic disease and formation of thrombi,hence result in hypoperfusion and infarction in posterior circulation cerebral tissue,which is potentially associated with posterior circulation cerebral vascular degeneration,vertebral artery dissection,cardiac disease and so on,also is a risk factor for posterior circulation infarction(PCI).The progresses of imaging researches revealing the association between VAH and PCI were reviewed in this article.
5.Clinical Observation of Acupuncture-activating Method for Deglutition Disorder Due to Pseudobulbar Palsy After Cerebral Stroke
Fawen ZHENG ; Ziming WU ; Jingtong MA ; Jiankang QIN
Shanghai Journal of Acupuncture and Moxibustion 2017;36(6):668-672
Objective To observe the clinical efficacy of ZHU Lian's acupuncture-activatingmethod in treating deglutition disorder due to pseudobulbar palsy after cerebral stroke.Method Sixty patients with deglutition disorder due to pseudobulbar palsy after cerebral stroke in acute stage were randomized into an observation group and a control group, 30 cases in each group. The two groups both received symptomatic neurological treatment. In the two groups, acupoints including Lianquan (CV23), Huiyan (Extra), Baihui (GV20), and bilateral Shuaigu (GB8), Wangu (GB12), Zhaohai (KI6), Lieque (LU7),and Yinlingquan (SP9) were selected. The observation group was intervened by ZHU Lian's acupuncture-activating method; the control group was intervened by ordinary needling method plus G6805 therapeutic appliance with sparse-dense wave for 20 min. Prior tothe treatment and after 10 treatment courses, the two groups were evaluated by using Videofluoroscopic Swallowing Study (VFSS) and Kubota's water drinking test for swallowing function.Result After 10 treatment courses, the total effective rate was 96.7%in the observation group versus 83.3% in the control group, and the between-group difference was statistically significant (P<0.05); the VFSS score in the observation group was significantly different from that in the control group (P<0.01); the water drinking test score in the observation group was significantly different from that in the control group (P<0.01). The results indicated that the therapeutic efficacy was more significant in the observation group compared to that in the control group.Conclusion ZHU Lian's acupuncture-activating method can produce a more significant efficacy in treating deglutition disorder due to pseudobulbar palsy after cerebral stroke in acute stage compared with sparse-dense-wave electroacupuncture.
6.Correlation between venous blood phase values detected by enhanced gradient echo T2 star weighted angiography sequence and neurological severity scale scores in mild and severe traumatic brain injury models of rabbits
Jingtong XIONG ; Jingwen ZHANG ; Jianlin WU ; Yuguo FANG
Chinese Journal of Radiology 2016;(2):132-137
Objective To explore the change of venous blood phase values detected by enhanced gradient echo T2 star weighted angiography(ESWAN)sequence in mild and severe traumatic brain injury (TBI) models of rabbits in diverse phases and investigate their association with neurological severity scale (NSS) scores. Methods Fifty-one New Zealand rabbits, which were randomly divided into control group (n=3) ,mild injured group (n=24) , and severe injured group (n=24) by random digital table method, underwent routine MRI and ESWAN sequence at the time points of baseline, 1 h, 6 h, 12 h, 24 h, 48 h, 72 h, 1 week, and 2 weeks after injury(3 rabbits per subgroup)respectively. Blood phase values in veins of interest were recorded. Observation of behavior characteristics and abnormalities, followed by NSS, was executed post injury, and the correlation between venous blood phase values and NSS scores was statistically analyzed. Two independent-samples t-test was applied to compare venous blood phase values of diverse measured veins in each group separately at every time ponits. One-way ANOVA was used to analyze venous blood phase values varying over time of measured vessels in each injured group. Least significant difference t-test was applied to compare blood phase values within the subgroups with each other at each time point. NSS scores of mild and severe injured groups were compared by two independent-samples t-test. Correlations between venous blood phase values and NSS scores in each vein and group was analyzed with Spearman correlation analysis. Results Blood phase values in veins of interest presented an inclination of descending, which was more obvious in severe injured groups than in the mild. Change of venous blood phase values over time featured continuing reduction in earlier phases which reached to the minimum in 24-48h, and then increased gradually, especially in VMV, VLV, DSS, and MDVB (P<0.05). ICV and DCV also showed similar trend. Values of NSS scores in the two group were (15.5 ± 3.1) and (33.2 ± 6.5) respectively (t=3.543,P=0.001). Blood phase values in most of the measured veins correlated with NSS scores (P<0.05) after 6h post injury, especially significant during 24-72h among all the measured veins(P<0.05). Higher accuracy and sensitivity were presented in the supratentorial or superficial veins compared with subtentorial or profound veins. Conclusions It is feasible to evaluate oxygen saturation of veins after TBI by measuring venous blood phase values on ESWAN images. The method is effective in the assessment of the degree of injury and clinical status, indicating a favorable application prospect.
7.Rotational animal brain injury models dynamically and quantitatively detected by diffusion tensor imaging correlated with β-amyloid precursor protein expression
Jingtong XIONG ; Jingwen ZHANG ; Jianlin WU ; Yuguo FANG ; Lin LIN
Chinese Journal of Radiology 2016;50(6):469-474
Objective To investigate dynamic regulation of quantifying fractional anisotropy (FA) and apparent diffusion coefficient (ADC) values in rotational brain injury models of rabbit using diffusion tensor imaging (DTI), and its correlation with β?amyloid precursor protein (β-APP). Methods Forty-two 6-month-old New Zealand rabbits were randomly divided into three groups, including control group (n=6), mild injured group (n=18) and severe injured group (n=18), and preformed on autonomous rapidly rotational brain injury device. The rotational angles of 45° for mild injured group and 90° for severe injured group were condemned, and MRI and pathology were conducted at 6 h, 12 h, 24 h, 48 h, 72 h and 1 week after injury (3 rabbits per subgroup). Routine sequences and DTI technique were performed on 3.0 T MRI. FA and ADC values in subcortical white matter, corpus callosum and brain stem were measured. Independent t?test was performed to evaluate the significance of the intergroup difference in FA and ADC values in mild and severe injured groups of verious brain regions by timing, one?way ANOVA was performed to evaluate its timing variation and its correlation with the number of the β-APP positive axons was analyzed by Pearson correlation analysis. Results FA and ADC values of the severe injured group were lower than that of the mild in most brain regions(P<0.05), and the difference in mild injured group was smaller than that in severe injured group. Both FA and ADC values in brain stem of the severe injured group were lower at 6 h after injury compared to mild injured group, which were sensitive to injury. Furthermore, FA and ADC values in each brain regions of mild and severe injured groups showed similar dynamic trends, namely gradually decreasing by time, and FA values were more sensitive to injury than ADC values. FA values in subcortical white matter and brain stem reduced in severe injured group at 6 h after injury compared with that before injury(P<0.05), and decreased in various brain area of both injured groups at 12 h after injury(P<0.05). Meanwhile, ADC values in all regions were declined in the severe injured group at 12 h after injury(P<0.05), and decreased in various regions in both injured groups at 24—48 h after injury(P<0.05)except for subcortical white matter in mild injured group. There were statistically negative correlations between FA and ADC values and the number of β-APP positive axons in 12—48 h after injury in most regions(P<0.05). Conclusions DTI can quantitatively detect and assess the pathological process in white matter and axons of TBI in earlier stage of the brain injury, and can be applied in evaluation and quantitative diagnose in these patients.
8.Expression and clinical significance of T regulatory cells in patients with gastric cancer
Jingtong WANG ; Yuanyuan ZHANG ; Tao PENG ; Yan WANG ; Jian YAO ; Shanshan ZHANG ; Jiang WU ; Yulan LIU
Chinese Journal of General Surgery 2008;23(11):850-852
Objective To investigate the proportion changes of the CD4 + CD25 + T cells and CD8 + CD28 - T cells in the peripheral blood of gastric cancer patients. Methods The proportions of CD4 + CD25 + T cells and CD8 + CD28 - T cells in the peripheral blood of gastric cancer patients were determined by flow cytometry. 30 patients with gastric cancer and 30 patients with chronic gastritis were enrolled in this study. Results Compared with the chronic gastritis controls, the proportion of the CD4 + CD25 + T cells in the peripheral blood of patients with gastric cancer (8.7%±1.3%) was not significantly changed. But the proportion of the CD8 + CD28 - T cell in the peripheral blood of patients with gastric cancer (30.3%±3.3%) was significantly higher than that of control group (20.3%±2.7%), P<0.05. There was no significant association between the proportions of T regulatory cells and the lymphomatic metastasis of gastric cancer. No relation was found between the proportions of T regulatory cells and pathological types of gastric cancer. Conclusion CD8 + CD28 - T ceils in peripheral blood may be of some value in the progression of gastric cancer.

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