1.Protective Effect of Huayu Jiedu Prescription Against PC12 Cell Damage Induced by Combination of NETs and OGD
Wuchaonan LIU ; Ruiyu HUANG ; Qianru ZENG ; Mengge ZHANG ; Yunhao YI ; Tao GAO ; Dingxiang LI ; Yihui DENG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(21):120-130
ObjectiveTo investigate the protective effect of Huayu Jiedu prescription against PC12 cell damage induced by a combination of neutrophil extracellular traps (NETs) and oxygen-glucose deprivation (OGD). MethodsFirst, dimethyl sulfoxide (DMSO) was used to induce the differentiation of the human promyelocytic leukemia cell line (HL-60) into neutrophil-like cells (dHL-60), and cell differentiation was assessed through Giemsa staining and flow cytometry. Next, phorbol myristate (PMA) was used to induce the formation of NETs in dHL-60 cells. The formation of NETs was assessed via SYTOX Green staining, and NETs were extracted by low-speed centrifugation. Finally, the NETs prepared through the conditioned medium were combined with OGD to induce a high-differentiation PC12 neuronal cell damage model through the co-culture method. This model was treated with the Huayu Jiedu prescription-containing serum. Cells were randomly allocated into a normal control group, a model (NETs + OGD) group, a traditional Chinese medicine (NETs + OGD + 5% Huayu Jiedu prescription-containing serum) group, and a NETosis inhibitor (100 mg·L-1 DNase I + NETs + OGD) group. The cell counting kit-8 (CCK-8) assay was used to evaluate cell viability and screen the optimal intervention concentrations of NETs and Huayu Jiedu Formula-containing serum, as well as the optimal intervention time for OGD. Enzyme-linked immunosorbent assay (ELISA) was adopted to measure the levels of myeloperoxidase-DNA complexes (MPO-DNA), citrullinated histone H3 (CitH3), and neutrophil elastase (NE). Western blot was employed to quantify the protein levels of CitH3 and cleaved Caspase-3. Terminal-deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay (TUNEL) staining and flow cytometry were employed to assess neuronal apoptosis. ResultsCompared with those in the normal control group, cells in the model group successfully differentiated into dHL60 cells after DMSO induction. Wright-Giemsa staining revealed that the cells in the model group had disappeared nucleoli, distinct reniform and lobulated changes of nuclei, highly condensed chromatin, and the cytoplasm containing pale purple fine granules. Flow cytometry showed that the mean fluorescence intensity (MFI) of CD11b in the model group was enhanced (P<0.01). Compared with those in the normal control group, dHL60 cells in the model group successfully generated NETs after PMA induction. SYTOX Green staining revealed loss of cell membrane integrity, with chromatin released from the cytoplasm to form abundant extracellular, cloud-like fibrillar structures, and a marked increase in fluorescence intensity (P<0.01). Through the CCK-8 assay for assessing the viability of highly differentiated PC12 cells, the optimal concentration for NETs was determined as 1 000 ng·L-1, the optimal concentration of Huayu Jiedu formula-containing serum as 5%, and the optimal duration for OGD as 2 h. Compared with the normal control group, the model group showed elevated levels of MPO-DNA, CitH3, and NE (P<0.01) and upregulated protein levels of CitH3 and cleaved Caspase-3 (P<0.01). Furthermore, TUNEL staining revealed an increased proportion of apoptotic cells (P<0.01) and flow cytometry showed an increased apoptosis rate (P<0.01) in the model group. Compared with the model group, the Huayu Jiedu formula-containing serum group and the NETosis inhibitor group exhibited declined levels of MPO-DNA, CitH3, and NE (P<0.05,P<0.01) and downregulated protein levels of CitH3 and cleaved Caspase-3 (P<0.01). In addition, TUNEL staining showed a decrease in the proportion of apoptotic cells (P<0.01), and flow cytometry revealed a decrease in the apoptosis rate (P<0.01). ConclusionHuayu Jiedu prescription may exert a protective effect against ischemic neuronal damage by inhibiting NET formation or alleviating NET-induced neurotoxicity
2.Protective Effect of Huayu Jiedu Prescription Against PC12 Cell Damage Induced by Combination of NETs and OGD
Wuchaonan LIU ; Ruiyu HUANG ; Qianru ZENG ; Mengge ZHANG ; Yunhao YI ; Tao GAO ; Dingxiang LI ; Yihui DENG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(21):120-130
ObjectiveTo investigate the protective effect of Huayu Jiedu prescription against PC12 cell damage induced by a combination of neutrophil extracellular traps (NETs) and oxygen-glucose deprivation (OGD). MethodsFirst, dimethyl sulfoxide (DMSO) was used to induce the differentiation of the human promyelocytic leukemia cell line (HL-60) into neutrophil-like cells (dHL-60), and cell differentiation was assessed through Giemsa staining and flow cytometry. Next, phorbol myristate (PMA) was used to induce the formation of NETs in dHL-60 cells. The formation of NETs was assessed via SYTOX Green staining, and NETs were extracted by low-speed centrifugation. Finally, the NETs prepared through the conditioned medium were combined with OGD to induce a high-differentiation PC12 neuronal cell damage model through the co-culture method. This model was treated with the Huayu Jiedu prescription-containing serum. Cells were randomly allocated into a normal control group, a model (NETs + OGD) group, a traditional Chinese medicine (NETs + OGD + 5% Huayu Jiedu prescription-containing serum) group, and a NETosis inhibitor (100 mg·L-1 DNase I + NETs + OGD) group. The cell counting kit-8 (CCK-8) assay was used to evaluate cell viability and screen the optimal intervention concentrations of NETs and Huayu Jiedu Formula-containing serum, as well as the optimal intervention time for OGD. Enzyme-linked immunosorbent assay (ELISA) was adopted to measure the levels of myeloperoxidase-DNA complexes (MPO-DNA), citrullinated histone H3 (CitH3), and neutrophil elastase (NE). Western blot was employed to quantify the protein levels of CitH3 and cleaved Caspase-3. Terminal-deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay (TUNEL) staining and flow cytometry were employed to assess neuronal apoptosis. ResultsCompared with those in the normal control group, cells in the model group successfully differentiated into dHL60 cells after DMSO induction. Wright-Giemsa staining revealed that the cells in the model group had disappeared nucleoli, distinct reniform and lobulated changes of nuclei, highly condensed chromatin, and the cytoplasm containing pale purple fine granules. Flow cytometry showed that the mean fluorescence intensity (MFI) of CD11b in the model group was enhanced (P<0.01). Compared with those in the normal control group, dHL60 cells in the model group successfully generated NETs after PMA induction. SYTOX Green staining revealed loss of cell membrane integrity, with chromatin released from the cytoplasm to form abundant extracellular, cloud-like fibrillar structures, and a marked increase in fluorescence intensity (P<0.01). Through the CCK-8 assay for assessing the viability of highly differentiated PC12 cells, the optimal concentration for NETs was determined as 1 000 ng·L-1, the optimal concentration of Huayu Jiedu formula-containing serum as 5%, and the optimal duration for OGD as 2 h. Compared with the normal control group, the model group showed elevated levels of MPO-DNA, CitH3, and NE (P<0.01) and upregulated protein levels of CitH3 and cleaved Caspase-3 (P<0.01). Furthermore, TUNEL staining revealed an increased proportion of apoptotic cells (P<0.01) and flow cytometry showed an increased apoptosis rate (P<0.01) in the model group. Compared with the model group, the Huayu Jiedu formula-containing serum group and the NETosis inhibitor group exhibited declined levels of MPO-DNA, CitH3, and NE (P<0.05,P<0.01) and downregulated protein levels of CitH3 and cleaved Caspase-3 (P<0.01). In addition, TUNEL staining showed a decrease in the proportion of apoptotic cells (P<0.01), and flow cytometry revealed a decrease in the apoptosis rate (P<0.01). ConclusionHuayu Jiedu prescription may exert a protective effect against ischemic neuronal damage by inhibiting NET formation or alleviating NET-induced neurotoxicity
3.Mechanism of Modified Erxian Decoction Regulating Perimenopausal Syndrome via SIRT1/Kisspeptin/GnRH Signaling Pathway
Ruiyu HUANG ; Fang LEI ; Wuchaonan LIU ; Jingjing YANG ; Qianru ZENG ; Shengping LUO ; Yanling CHEN ; Mengge ZHANG ; Fanshun SHEN ; Yihui DENG ; Dingxiang LI
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(22):51-62
ObjectiveTo explore the regulation of hypothalamic-pituitary-gonadal (HPG) axis by modified Erxian decoction in rats with perimenopausal syndrome (PMS) and to further analyze the expression of proteins related to the silent information regulator 1 (SIRT1)/hypothalamic kisspeptin (Kisspeptin)/gonadotropin-releasing hormone (GnRH) signaling pathway in the arcuate nucleus region (ARC) of the hypothalamus, so as to reveal the potential target of action and molecular biological mechanism of modified Erxian decoction for the treatment of perimenopausal syndrome. MethodsAn animal model was established via the incomplete castration method, with successful modeling confirmed by the exfoliated cervical cell smear method. The 48 rats were divided into six groups based on the randomization principle after successful modeling, including a sham operation group, a model group, an estradiol valerate group (0.09 mg∙kg-1∙d-1), high-, medium-, and low-dose modified Erxian decoction groups (7.614, 3.807,1.903 5 g∙kg-1∙d-1), with 8 rats in each group. The estradiol valerate group and the high-, medium- and low-dose modified Erxian decoction groups were continuously administered by gavage for 28 days, and the indicators were detected 24 hours after the last administration. Body weights and uterine indices were measured. The pathological changes of the uterus were observed by hematoxylin-eosin (HE) staining. Enzyme-linked immunosorbent assay (ELISA) was performed to measure the levels of estradiol (E2), follicle-stimulating hormone (FSH), luteinizing hormone (LH), and gonadotropin-releasing hormone (GnRH). Real-time quantitative polymerase chain reaction (Real-time PCR) and Western blot were used to determine the expression levels of SIRT1, Kisspeptin, kisspeptin receptor (GPR54), and GnRH in the ARC region of the hypothalamus and gonadotropin-releasing hormone receptor (GnRH-R) in pituitary. ResultsCompared with the sham operation group, rats in the model group had a significantly increased body weight (P0.01), reduced wet weight and index of uterus (P0.01), endometrial thinning or atrophy, glandular atrophy, and a decreasing number of glands. Additionally, serum levels of E2 and the expression of SIRT1 in the ARC region of the hypothalamus significantly decreased (P0.01). Serum levels of FSH, LH, and GnRH, the expression of Kisspeptin, GPR54, and GnRH in the ARC region of the hypothalamus, and GnRH-R in pituitary significantly increased (P0.01). Compared with the model group, the estradiol valerate group and the high-, medium-dose modified Erxian decoction groups had significantly reduced body weight, serum levels of FSH, LH, and GnRH, and expression of Kisspeptin, GPR54, and GnRH in the ARC region of the hypothalamus and GnRH-R in pituitary (P0.05, P0.01) and significantly increased wet weight and index of uterus, serum level of E2, and expression of SIRT1 in the ARC region of the hypothalamus (P0.05, P0.01). In addition, they showed thickened endometrium, increased number of endometrial glands, and improved glandular atrophy. ConclusionModified Erxian decoction regulates the function of the HPG axis through multi-targets, and its mechanism of action may be related to the up-regulation of the expression of SIRT1 in the ARC region of the hypothalamus, the inhibition of the over-activation of the Kisspeptin/GnRH signaling pathway, the regulation of the expression of GnRH-R in the pituitary, the restoration of secretion balance of gonadotropins, and the elevation of the estrogen level. This study provides an experimental basis for the interpretation of the scientific connotation of modified Erxian decoction in the treatment of perimenopausal syndrome and a theoretical reference for the development of a novel therapeutic strategy based on the SIRT1/Kisspeptin/GnRH pathway.
4.Comparison of clinicopathological and MRI imaging features between ductal carcinoma in situ with microinfiltration and ductal carcinoma in situ of the breast
Hong'en LI ; Li ZHANG ; Xia WANG ; Xing LIN ; Wei WANG ; Yihui ZENG
Chinese Journal of Oncology 2025;47(8):726-733
Objective:To investigate the differences in the clinicopathological and magnetic resonance imaging (MRI) imaging features between ductal carcinoma in situ (DCIS) and ductal carcinoma in situ with microinfiltration (DCIS-MI) of the breast, and to clarify the risk factors for the development of DCIS-MI.Methods:Forty-four patients diagnosed with DCIS and 21 patients diagnosed with DCIS-MI by postoperative pathology at Guangdong Maternal and Child Health Hospital from November 2017 to November 2022 were included, and the clinicopathological and preoperative breast MRI data of these patients were retrospectively collected. The patients' MRI images were categorized and diagnosed with reference to the Breast Imaging Reporting and Data System (BI-RADS) criteria. The χ2 test or Fisher exact probability method was used to compare the differences in the clinicopathological and MRI imaging characteristics between the two groups of patients, and generalized linear model analysis was used to clarify the influencing factors of DCIS-MI.Results:The differences in the histologic grading, estrogen receptor (ER) expression, progesterone receptor (PR) expression, human epidermal growth factor receptor 2 (HER-2) expression, Ki-67, and molecular typing between patients in the DCIS and DCIS-MI groups were statistically significant (all P<0.05). The results of generalized linear model analysis showed that Ki-67 expression and specific molecular typing (Luminal B and triple-negative types) were significantly associated with the risk of developing DCIS-MI ( P<0.05). Breast fibroglandular tissue density, lesion type, background parenchymal enhancement, type of time-intensity curves (TICs), distribution of non-mass enhancement, non-mass enhancement internal enhancement characteristics, mass morphology, mass boundary, mass enhancement mode, and other MRI imaging features were not statistically significant (all P>0.05).The MRI diagnostic accuracy of the DCIS group and the DCIS-MI group was 77.3% (34/44) and 95.2% (20/21), respectively, and the difference in the MRI BI-RADS classification of the patients in the two groups was not statistically significant ( P=0.227). Conclusions:There was no significant difference in the breast MRI imaging characteristics between patients in the DCIS and DCIS-MI groups. Patients in the DCIS-MI group were more likely to present with high histologic grades, negative ER, negative PR, positive HER-2, high Ki-67 expression, HER-2 overexpression, and triple-negative phenotypes. The association between Ki-67 expression and specific molecular typing (Luminal B and triple-negative phenotypes) and the risk of developing DCIS-MI risk were correlated.
5.Comparison of clinicopathological and MRI imaging features between ductal carcinoma in situ with microinfiltration and ductal carcinoma in situ of the breast
Hong'en LI ; Li ZHANG ; Xia WANG ; Xing LIN ; Wei WANG ; Yihui ZENG
Chinese Journal of Oncology 2025;47(8):726-733
Objective:To investigate the differences in the clinicopathological and magnetic resonance imaging (MRI) imaging features between ductal carcinoma in situ (DCIS) and ductal carcinoma in situ with microinfiltration (DCIS-MI) of the breast, and to clarify the risk factors for the development of DCIS-MI.Methods:Forty-four patients diagnosed with DCIS and 21 patients diagnosed with DCIS-MI by postoperative pathology at Guangdong Maternal and Child Health Hospital from November 2017 to November 2022 were included, and the clinicopathological and preoperative breast MRI data of these patients were retrospectively collected. The patients' MRI images were categorized and diagnosed with reference to the Breast Imaging Reporting and Data System (BI-RADS) criteria. The χ2 test or Fisher exact probability method was used to compare the differences in the clinicopathological and MRI imaging characteristics between the two groups of patients, and generalized linear model analysis was used to clarify the influencing factors of DCIS-MI.Results:The differences in the histologic grading, estrogen receptor (ER) expression, progesterone receptor (PR) expression, human epidermal growth factor receptor 2 (HER-2) expression, Ki-67, and molecular typing between patients in the DCIS and DCIS-MI groups were statistically significant (all P<0.05). The results of generalized linear model analysis showed that Ki-67 expression and specific molecular typing (Luminal B and triple-negative types) were significantly associated with the risk of developing DCIS-MI ( P<0.05). Breast fibroglandular tissue density, lesion type, background parenchymal enhancement, type of time-intensity curves (TICs), distribution of non-mass enhancement, non-mass enhancement internal enhancement characteristics, mass morphology, mass boundary, mass enhancement mode, and other MRI imaging features were not statistically significant (all P>0.05).The MRI diagnostic accuracy of the DCIS group and the DCIS-MI group was 77.3% (34/44) and 95.2% (20/21), respectively, and the difference in the MRI BI-RADS classification of the patients in the two groups was not statistically significant ( P=0.227). Conclusions:There was no significant difference in the breast MRI imaging characteristics between patients in the DCIS and DCIS-MI groups. Patients in the DCIS-MI group were more likely to present with high histologic grades, negative ER, negative PR, positive HER-2, high Ki-67 expression, HER-2 overexpression, and triple-negative phenotypes. The association between Ki-67 expression and specific molecular typing (Luminal B and triple-negative phenotypes) and the risk of developing DCIS-MI risk were correlated.
6.Urinary Metabolomics Aanlysis of Differences in Effect of Aconiti Coreani Radix and Typhonii Rhizoma on Gerbils with Stroke
Liting ZHOU ; Wanting ZENG ; Ru JIA ; Huiying XU ; Yihui DING ; Hao DONG ; Haowen MA ; Yang QU ; Qian CAI
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(8):157-166
ObjectiveTo investigate the effects of Aconiti Coreani Radix and Typhonii Rhizoma on the urinary metabolites of gerbils with stroke by non-targeted metabolomics technique, and then to clarify the mechanism of the two, as well as their similarities and differences. MethodTwenty-four gerbils were randomly divided into control group(CG), model group(MG), Aconiti Coreani Radix group(RA) and Typhonii Rhizoma group(RT). Except for the CG, ischemic stroke model was constructed using right unilateral ligation of gerbil carotid artery in the remaining groups. Except for the CG and MG, rats in the other groups received whole powder suspension(0.586 mg·g-1) was administered for 14 days. The neurological deficit in each group was scored by Longa scoring on days 0, 3, 7 and 14. After the end of administration, the serum, brain tissue and urine of gerbils in each group were collected, and the rate of cerebral infarction was detected by 2,3,5-triphenyltetrazolium chloride(TTC), and the levels of interleukin(IL)-6, tumor necrosis factor(TNF)-α, malondialdehyde(MDA), superoxide dismutase(SOD), glutathione(GSH), and nitric oxide(NO) in serum and brain tissue were determined by enzyme-linked immunosorbent assay(ELISA). The urine metabolomics of gerbils in each group was studied by ultra performance liquid chromatography-quadrupole-electrostatic field orbitrap high resolution mass spectrometry(UPLC-Q-Orbitrap-MS), and the data were processed by multivariate statistical analysis, and differential metabolites were screened based on value of variable importance in the projection(VIP) of the first principal component>1 and t-test P<0.05. Metabolic pathway analysis of the screened differential metabolites was performed using Kyoto Encyclopedia of Genes and Genomes(KEGG) database and Metaboanalyst 5.0. ResultCompared with the CG, the neurological deficit score was significantly increased in the MG(P<0.05), compared with the MG, the neurological deficit scores in the RA and RT were significantly reduced after 7 d and 14 d(P<0.05). Compared with the CG, the rate of cerebral infarction was significantly increased in the MG(P<0.05), compared with the MG, the rates of cerebral infarction in the RA and RT were significantly reduced(P<0.05). Compared with the CG, the levels of IL-6, TNF-α, and MDA in the serum and brain tissue of gerbils from the MG were significantly increased(P<0.05), and the levels of SOD, GSH and NO were significantly reduced(P<0.05). Compared with the MG, Aconiti Coreani Radix and Typhonii Rhizoma could down-regulate the levels of IL-6, TNF-α and MDA, and up-regulated the levels of SOD, GSH and NO. A total of 112 endogenous differential metabolites were screened by urine metabolomics, of which 16 and 26 metabolites were called back by Aconiti Coreani Radix and Typhonii Rhizoma, and could be used as potential biomarkers for both treatments in stroke gerbils, respectively. The results of the pathway analysis showed that both Aconiti Coreani Radix and Typhonii Rhizoma had regulatory effects on arginine and proline metabolism, pyrimidine metabolism, and aminoacyl-tRNA biosynthesis. In addition, Aconiti Coreani Radix could also regulate riboflavin metabolism, Typhonii Rhizoma could also regulate purine metabolism, glycine, serine and threonine metabolism, arachidonic acid metabolism, biosynthesis of pantothenate and coenzyme A, and β-alanine metabolism. ConclusionBoth Aconiti Coreani Radix and Typhonii Rhizoma have better therapeutic effects on stroke, with Aconiti Coreani Radix having stronger effects. From the metabolomics results, the main metabolic pathways regulated by Aconiti Coreani Radix involve amino acid metabolism, oxidative stress and so on, while Typhonii Rhizoma mainly involve amino acid metabolism, lipid metabolism, energy metabolism, etc.
7.Research progress of microRNA-mediated immune function of CD4+T cells subsets in immune thrombocytopenia
Hui SONG ; Yihui GUO ; Jiawei XU ; Qing ZENG ; Weimin CHENG
Chinese Journal of Immunology 2024;40(3):663-667
MicroRNA(miRNA)is a kind of small non-coding single stranded RNA that can participate in multiple biological processes.It also plays an important role in regulating the immune function of the body.Immune thrombocytopenia(ITP)is an autoim-mune disease,whose cause and deterioration are closely related to miRNA regulates immune function of CD4+T cells subsets.In ITP patients,different expression of miRNA can affect the immune function of CD4+T cells subsets,which causes not only unbalanced ex-pression of Th1/Th2,Th17/Treg and excessive differentiation of TFH,but also abnormal cytokine secretion furthermore.This paper summarizes the unbalanced mechanism of miRNA regulating immune function of CD4+T cells subsets in ITP,so as to provide inspira-tion for exploring the immunology and immunotherapy of ITP.
8.Application of rice bag in 1.5T MR imaging in maxillofacial
Hong'en LI ; Yuelong LI ; Hanmin XIE ; Peifeng LÜ ; Chengwei LI ; Yihui ZENG
Journal of Practical Radiology 2024;40(2):319-322
Objective To investigate the application value of rice bag in 1.5T MR imaging of maxillofacial frequency selective saturated fat suppression(FS)sequence.Methods Forty-one patients with maxillofacial MR examination were collected.Rapid fast spin echo(FSE)FS T2 routine scan and rice bag FSE FS T2 special scan were used to evaluate and analyze the sequence images of the two kinds by two experienced associate chief physicians in the department according to the Likert 7-point scoring system.The evaluation included FS,magnetic sensitive artifact,motion artifact,and overall image quality.Wilcoxon sign rank sum test was used to analyze the difference of image quality scores,and Kappa consistency test was used to analyze the correlation between two viewers.Results All the 41 patients completed the examination,and the FSE FS T2 plus rice bag group had significantly higher FS,magnetic sensitive artifact,motion artifact and overall image quality than the conventional FSE FS T2 group,with statistical significance(P<0.01).Kappa consistency was good in correlation analysis between the two viewers(Kappa value=0.737-0.877,P<0.01).Conclusion The application of rice bag in the 1.5T MR maxillofacial FS sequence scan has the advantages of good image quality,good FS effect and few artifacts,which can better display the maxillofacial anatomical structure and lesions,and has important clinical application value for the diagnosis of maxillofacial diseases.
9.The diagnostic value of ultrasound combined with MRI in fetal urinary system abnormalities
Hong'en LI ; Simin HUANG ; Dawei TANG ; Siyue WANG ; Chengwei LI ; Yihui ZENG
Journal of Practical Radiology 2024;40(5):768-771
Objective To investigate the application value of ultrasound combined with MRI in the diagnosis of fetal urinary system abnormalities.Methods The clinical data of 647 cases of fetal urinary system abnormalities suspected by prenatal ultrasound were ana-lyzed retrospectively.All pregnant women underwent MRI examination within 2 days after ultrasound examination.Using pathology and follow-up as the gold standard,the accuracy and imaging features of ultrasound and MRI in detecting fetal urinary system abnor-malities were analyzed.Results Among 600 cases confirmed by pathology and follow-up,the accuracy of MRI,ultrasound and com-bined diagnosis of fetal urinary system abnormalities were 96.50%,96.67%,97.00%,sensitivity were 95.28%,96.23%,98.11%,and specificity were 96.76%,96.76%,96.76%,respectively.There were no significant differences in accuracy,sensitivity,specificity,positive predictive value and negative predictive value between MRI,ultrasound and combined diagnosis(P>0.05).The area under the curve(AUC)of MRI,ultrasound and combined diagnosis of fetal urinary system abnormalities were 0.960,0.965 and 0.974,respectively,with no statistical significance(P>0.05).Conclusion Compared with prenatal ultrasound,MRI also has a higher accuracy(96.50%)in the diagnosis of fetal urinary system abnormalities,and can be used as an important supplement to prenatal ultrasound.
10.Effect and Mechanism of Qixian Tongluo Formula on Contralateral Corticospinal Tract Remodeling and Motor Functional Recovery in Rats with Cerebral Infarction
Shengqiang ZHOU ; Dahua WU ; Bo LI ; Yanjun CHEN ; Jia HUANG ; Qi WANG ; Wen ZENG ; Lingjuan TAN ; Yihui DENG ; Fang LIU
Traditional Chinese Drug Research & Clinical Pharmacology 2024;35(9):1319-1328
Objective To observe the effect of Qixian Tongluo Formula on contralateral corticospinal tract(CST)remodeling and motor functional recovery in rats with cerebral infarction,and to explore its potential molecular mechanism from the perspective of regulating factors related to never remodeling.Methods The rat middle cerebral artery occlusion(MCAO)model was established by silk thread ligation.Fifty model rats were randomly divided into model group,citicoline group(0.054 g·kg-1),Qixian Tongluo Formula low-,medium-and high-dose(7.83,15.66,31.32 g·kg-1)groups,and sham operation group,with 10 rats in each group.The intervention administration was started on the 3rd day after operation once a day for 26 consecutive days.On the 3rd,14th and 28th day after operation,the gross motor function was evaluated by Longa score,and the fine motor function was evaluated by beam-walking test(BWT)score.The contralateral motor cortex was injected with the nerve tracer biotin dextran amine(BDA)on the 14 th day after operation to anterogradely trace the CST.On the 28th day after operation,the expression of axonal growth associated protein-43(GAP-43)and BDA positive fibers in the contralateral motor cortex and cervical spinal cord were detected by immunohistochemistry.The co-localization areas of BDA positive fibers and presynaptic marker protein vesicular glutamate transporter 1(VGLUT1)in the cervical spinal cord gray matter were detected by immunofluorescence.The expressions of brain-derived neurotrophic factor(BDNF),glial cell-derived neurotrophic factor(GDNF),nerve growth factor(NGF)and nerve remodeling-associated inhibitory factor[Nogo-A,oligodendrocyte myelin glycoprotein(OMgp)and myelin-associated glycoprotein(MAG)]in the contralateral motor cortex were detected by Western Blot.Pearson correlation analysis was used to analyze the correlation between Longa score or BWT score and BDA/VGLUT1 co-localization area,respectively.Results Compared with the sham operation group,rats in the model group had obvious symptoms of motor function deficits,and the Longa scores were significantly increased(P<0.01)and the BWT scores were significantly decreased(P<0.05,P<0.01)at each time point.The expression of GAP-43 in the contralateral motor cortex and cervical spinal cord was up-regulated(P<0.05),the number of edge-crossing fibers from the posterior funiculus in cervical cord was increased(P<0.05),the co-localization area of BDA/VGLUT1 in the gray matter of the cervical spinal cord was increased(P<0.05),the expressions of BDNF,GDNF and NGF in the contralateral motor cortex were up-regulated(P<0.05),while the expressions of Nogo-A,OMgp and MAG were down-regulated(P<0.05).Compared with the model group,the Longa scores in each administration group on the 14th and 28th day after MCAO operation were significantly decreased(P<0.01),the BWT scores were significantly increased(P<0.01),the expression of GAP-43 in the contralateral motor cortex and cervical spinal cord was significantly up-regulated(P<0.01).The number of edge-crossing fibers from the posterior funiculus in cervical cord was significantly increased(P<0.01),the co-localization area of BDA/VGLUT1 in the gray matter of the cervical spinal cord was significantly increased(P<0.01).The expressions of BDNF,GDNF and NGF in the contralateral motor cortex were significantly up-regulated(P<0.01,P<0.05),while the expressions of Nogo-A,OMgp and MAG were significantly down-regulated(P<0.05),and the most significant effect was observed in the high dose group.The Longa score was negatively correlated with the co-localization area of BDA/VGLUT1(r=-0.89,P<0.01),and the BWT score was positively correlated with the co-localization area of BDA/VGLUT1(r=0.84,P<0.01).Conclusion Qixian Tongluo Formula can improve motor function through promoting contralateral CST remodeling in MCAO rats after cerebral infarction,and the molecular mechanism may be related to the regulation of the expression of nerve remodeling-associated factor in the contralateral motor cortex.

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