1.Effect of laminin subunit α3 on epithelial-mesenchymal transition, invasion, and metastasis abilities of pancreatic cancer
Nenghong YANG ; Likun REN ; She TIAN ; Min HAN ; Zhu LI ; Yuxiang ZHAO ; Peng LIU
Journal of Clinical Hepatology 2025;41(2):322-332
ObjectiveTo investigate the effect of laminin subunit α3 (LAMA3) on the epithelial-mesenchymal transition (EMT), invasion, and metastasis abilities of pancreatic cancer (PC). MethodsA comprehensive analysis was performed for tumor- and EMT-related databases to identify the EMT genes associated with PC, especially LAMA3. The methods of qRT-PCR and Western blot were used to measure the expression level of LAMA3 in PC tissue and cell lines; immunofluorescence assay was used to determine the localization of LAMA3 in PANC-1 cells; Transwell assay was used to investigate the effect of LAMA3 on the invasion and migration abilities of PC cells. The t-test was used for comparison of continuous data between groups. ResultsThe analysis of the TCGA database identified 3 EMT-related oncogenes for PC, i.e., LAMA3, AREG, and SDC1. The LASSO-Cox regression model showed that LAMA3 had the most significant impact on the prognosis of PC (risk score=0.256 1×LAMA3+0.043 1×SDC1+0.071 4×AREG). The Cox model and nomogram showed that the high expression of LAMA3 was an independent risk factor for the poor prognosis of PC (hazard ratio=1.32, 95% confidence interval: 1.07 — 1.62, P<0.01). Experimental results showed that there was a significant increase in the expression of LAMA3 in pancreatic cancer tissue compared with the normal pancreatic tissue. Compared with the HPDE cell line, there were varying degrees of increase in the expression of LAMA3 in pancreatic cancer AsPC-1, BxPC-3, PANC-1, MIA PaCa-2, and SW1990 cell lines, with the highest expression level in PANC-1 cells. The enrichment analysis showed that LAMA3 was associated with the biological processes and signaling pathways such as EMT, collagen metabolism, extracellular matrix degradation, the TGF-β pathway, and the PI3K pathway. After the knockdown of LAMA3, there were significant reductions in the expression levels of N-Cadherin, Vimentin, and Snail, while there was a significant increase in the expression level of E-Cadherin. Transwell assay showed that there were significant reductions in the invasion and migration abilities of PANC-1 cells after the knockdown of LAMA3. ConclusionLAMA3 is highly expressed in PC and can promote the EMT, invasion, and migration of PC cells, and therefore, LAMA3 may be used as a novel diagnostic marker and a new therapeutic target for PC.
2.An Enhancer LncRNA Regulates NFE2 Expression and Proliferation in Human Leukemic K562 Cells
Yan-Fei LU ; Song-Ya QU ; Jing-Jing ZHU ; Chao LIU ; Jian WANG ; Bing-She HAN ; Jun-Fang ZHANG
Progress in Biochemistry and Biophysics 2024;51(1):190-201
ObjectiveTranscription factor NFE2 was observed abnormal expression in myeloproliferative neoplasm (MPN) patients. However, how NFE2 is transcriptionally regulated remains ambiguous. This study aims to explore the elements and molecular mechanisms involved in the transcriptional regulation of NFE2. MethodsActive enhancers were predicted by public NGS data and conformed experimentally via dual luciferase reporter assay. After that, PRO-seq and GRO-seq data was used to detect enhancer RNAs transcribed from these enhancers. RACE was utilized to clone the full length enhancer RNA (eRNA) transcripts, and RT-qPCR was used to measure their expression in different leukemia cell lines as well as the transcript levels during induced differentiation. Finally, to investigate the molecular function of the eRNA, overexpression and knockdown of the eRNA via lentivirus system was performed in K562 cells. ResultsWe identified three enhancers regulating NFE2 transcription, which located at -3.6k, -6.2k and +6.3k from NFE2 transcription start site (TSS) respectively. At the -3.6k enhancer, we cloned an eRNA transcript and characterized that as a lncRNA which was expressed and located in the nucleus in three types of leukemia cell lines. When this lncRNA was overexpressed, expression of NFE2 was upregulated and decreases of K562 cell proliferation and migration ability were observed. While knocking down of this lncRNA, the level of NFE2 decreases correspondingly and the proliferation ability of K562 cells increases accordingly. ConclusionWe identified an enhancer lncRNA that regulates NFE2 transcription positively and suppresses K562 cell proliferation.
3.Heterologous expression and product identification of diterpene synthase involved in the biosynthesis of brasilicardin A
Xiang-yu GE ; Guang-xin ZHOU ; Na XIONG ; Zi-han LU ; Xin-yu MI ; Zhi-xiang ZHU ; Xiao LIU ; Xiao-hui WANG ; Juan WANG ; She-po SHI
Acta Pharmaceutica Sinica 2024;59(7):2161-2170
Brasilicardin A, a diterpene glycoside isolated from pathogenic actinomycete
4.Application value of early enteral nutrition management based on severe feeding process in patients with acute exacerbation of chronic obstructive pulmonary disease after mechanical ventilation
Chinese Journal of Practical Nursing 2024;40(5):344-349
Objective:To explore the effects of early enteral nutrition (EEN) management based on severe feeding process on nutritional status and gastrointestinal tolerance in patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD) after mechanical ventilation.Methods:In the quasi trial study, 30 patients with AECOPD undergoing mechanical ventilation between January 2020 and June 2021 and other 30 patients undergoing mechanical ventilation between July 2021 and December 2022 in Liu′an Hospital of Traditional Chinese Medicine were enrolled as control group and observation group, respectively. The control group was given routine EEN intervention, while the observation group was given EEN management based on severe feeding process. The intakes of calorie and proteins within 7 d after intervention were compared between the two groups. The gastrointestinal tolerance and prognosis were recorded. The levels of hemoglobin, albumin and prealbumin were compared between the two groups before intervention and at 7 d after intervention.Results:In the control group, there were 17 males and 13 females, with age of (66.37±6.09) years old. In the observation group, there were 16 males and 14 females, with age of (65.49 ± 6.35) years old. After intervention, intakes of calorie and proteins within 7 d in the observation group were (3 513.62 ± 94.56) kJ/d and (51.06 ± 5.03) g/d, more than those in the control group (2 859.41 ± 87.23) kJ/d, (36.78 ± 3.14) g/d, the differences were statistically significant ( t=27.85, 13.19, both P<0.05). There were no significant difference in the levels of hemoglobin, albumin and prealbumin before intervention between the two groups (all P>0.05). After intervention, levels of albumin, hemoglobin and prealbumin were (37.16 ± 3.42), (135.43 ± 12.64) g/L and (218.54 ± 15.38) mg/L in the observation group, higher than those in the control group (34.25 ± 4.01), (127.51 ± 11.12) g/L and (201.33 ± 15.04) mg/L, the differences were statistically significant ( t=3.02, 2.58, 4.38, all P<0.05). The incidence rates of reflux aspiration and enteral nutrition interruption in the observation group were 6.67% (2/30), 6.67% (2/30), lower than those in the control group 26.67% (8/30), 30.00% (9/30), the differences were statistically significant ( χ2=4.32, 5.46, both P<0.05). The mechanical ventilation time and stay time in ICU in the observation group were (7.62 ± 1.35), (11.17 ± 2.04) d, shorter than those in the control group (8.57 ± 1.01), (12.83 ± 2.19) d, the differences were statistically significant ( t=3.09, 3.04, both P<0.05). Conclusions:EEN management based on severe feeding process can improve nutritional status, gastrointestinal tolerance and prognosis of AECOPD patients after mechanical ventilation.
5.Study on protective effects and mechanism of ziyuglycoside Ⅰ on acute lung injury in sepsis rats
Daiqin BAO ; Yiyan LIU ; Zisen ZHANG ; Han SHE ; Lei TAN ; Tao LI ; Qingxiang MAO ; Liangming LIU
China Pharmacy 2023;34(5):537-543
OBJECTIVE To investigate the protective effects and mechanism of ziyuglycoside Ⅰ on acute lung injury in sepsis rats based on network pharmacology, and conduct experimental verification. METHODS The network pharmacology was used to predict the potential target of ziyuglycoside Ⅰ in the treatment of acute lung injury following sepsis. The rat model of sepsis was reproduced by cecum ligation and puncture for experimental verification. Totally 192 SD rats were randomly divided into the sham operation group (Sham group), sepsis group (Sep group), conventional therapy group (CT group) and ziyuglycoside Ⅰ group (Zg Ⅰ group), respectively. Sham group and Sep group were given sterile normal saline, and CT group and ZgⅠ group were given relevant volume of Ringer’s solution and ziyuglycoside Ⅰ. The arterial blood gas, serum inflammatory factors, lung wet/dry mass ratio, pathological changes of lung tissue, pulmonary vascular permeability, the expressions of pulmonary vein tight junction protein 1 (ZO-1) and vascular endothelial cadherin (VE-cadherin) protein and 72-hour survival were observed in each group. RESULTS Results of network pharmacology showed that there were 47 potential targets of ziyuglycoside Ⅰ in the treatment of sepsis. The results of gene ontology function enrichment analysis and Kyoto encyclopedia of genes and genomes pathway enrichment analysis showed that the mechanism could 598486924@qq.com be correlated with biological processes such as positive regulation of reactive oxygen species metabolism, wound healing, regulation of endothelial cell proliferation, cell activation, blood vessel development, response to oxidative stress, etc., and with signaling pathway such as apoptosis, tight junction, HIF-1 signaling pathway, etc. The results of experimental verification showed that compared with Sham group, pH value and the level of partial arterial oxygen pressure were decreased significantly in Sep group (P<0.05), while the level of partial pressure of carbon dioxide, serum levels of tumor necrosis factor α, interleukin 6 were increased significantly (P<0.05); the ratio of lung wet/dry mass was increased significantly (P<0.05); the protein expressions of ZO-1 and VE-cadherin were decreased significantly (P<0.05); 72 h survival rate decreased,the survival time was significantly shortened (P<0.05); the results of pathological observation of lung tissue showed that the rats’ alveoli were extensively ruptured, the alveolar wall was thickened and accompanied with edema, and there was obvious inflammatory cell infiltration; the results of pulmonary vascular permeability observation showed that the lung surface of rats was dark, with a large amount of Evans blue exudation, and the left lower lung was obviously dark blue. Compared with Sep group, the levels of above indexes almost were reversed significantly in CT group and ZgⅠ group (P<0.05); the lung histopathology and pulmonary vascular permeability were significantly improved, and the recovery degree of ZgⅠ group was greater than that of CT group, which was close to the results of Sham group. CONCLUSIONS Ziyuglycoside Ⅰ can significantly reduce inflammatory reaction and acute lung injury in septic rats, which is related to vascular function and tight junction signal pathway.
6.Osteosarcoma with bone metastasis or pulmonary metastasis show distinct genomic manifestations
Zhenyu CAI ; Yanchun SHE ; Lu XIE ; Han WANG ; Zhiye DU ; Yuan LI ; Tingting REN ; Jie XU ; Xin SUN ; Kunkun SUN ; Danhua SHEN ; Xiaodong TANG ; Wei GUO
Chinese Journal of Orthopaedics 2023;43(9):581-590
Objective:To investigate the genomic manifestation and pathogenesis of osteosarcoma with different relapse pattens, which were respectively initially presented with bone metastasis or pulmonary metastasis.Methods:From May 1, 2021 to October 1, 2021, 38 fresh tumor specimens and some paraffin-embedded specimens of high-grade osteosarcoma were collected in Peking University People's Hospital, including 29 males and 9 females, aged 19.6±2.2 years (range, 6-61 years). Among the 38 cases, 12 cases had initial bone metastasis (group A) and 26 cases had initial lung metastasis (group B), of which 15 cases (40%, 15/38) had paired specimens of primary and metastatic lesions. Based on Illumina NovaSeq 6000, we analyzed whole-exome sequencing (WES) as well as transcriptome for osteosarcoma with paired samples in different relapse patterns. During all their treatment courses, we also collected their paired samples to reveal these tumors' evolution. We sought to redefine disease subclassifications for osteosarcoma based on genetic alterations and correlate these genetic profiles with clinical treatment courses to elucidate potential evolving cladograms.Results:We found that osteosarcoma in group A mainly carried single-nucleotide variations (83%, 10/12), displaying higher tumor mutation burden [4.9 (2.8, 12.0) & 2.4 (1.4, 4.5), P=0.010] and neoantigen load [743.0 (316.5, 1,034.5) & 128.5 (49.0, 200.5), P=0.003], while those in group B mainly exhibit structural variants (58%, 15/26). The mutation spectrum showed that there was a significant difference in age-related gene imprinting 1 between the bone metastasis group and the lung metastasis group ( P=0.005). Samples were randomly selected from group A (3 patients) to investigate immunologic landscape by multiplex immunohistochemistry, from which we noticed tertiary lymphatic structure from one patient from group A. High conservation of reported genetic sequencing over time was found in their evolving cladograms. Conclusion:Osteosarcoma with mainly single-nucleotide variations other than structural variants might exhibit biological behavior predisposing toward bone metastases with older in age as well as better immunogenicity in tumor microenvironment.
7.Characteristics of plasma Epstein-Barr virus DNA in children with primary infection.
Yu Han ZHANG ; Fei LI ; Yuan Yuan ZHOU ; Peng SHI ; Ling Feng CAO ; Jian She WANG ; Jun SHEN
Chinese Journal of Pediatrics 2023;61(3):245-249
Objective: To explore the characteristics of plasma Epstein-Barr virus (EBV) DNA in primary infection in pediatric cases. Methods: The laboratory and clinical data of 571 children diagnosed with EBV primary infection in Children's Hospital of Fudan University during September 1st, 2017 to September 30th, 2018 were retrospectively analyzed. According to the results of plasma EBV DNA, they were divided into positive group and negative group. According to the EBV DNA, they were devided into high plasma virol load group and low plasma virol load group. The Chi-square test, Wilcoxon rank sum test were used to compare the differences between groups. Results: Among the 571 children with EBV primary infection, 334 were males and 237 were females. The age of first diagnosis was 3.8 (2.2, 5.7) years. There were 255 cases in positive group and 316 cases in negative group. The percentage of cases with fever,hepatomegaly and (or) splenomegaly, elevated transaminase in the positive group were higher than those in the negative group (235 cases (92.2%) vs. 255 cases (80.7%), χ2=15.22, P<0.001; 169 cases (66.3%) vs. 85 cases (26.9%), χ2=96.80, P<0.001; and 144 cases (56.5%) vs. 120 cases (38.0%), χ2=18.27, P<0.001; respectively).In the positive group, 70 cases were followed up for 46 (27, 106) days, 68 cases (97.1%) turned negative within 28 days, with the exception of 2 cases (2.9%) developed chronic active EBV infection by follow-up revision.There were 218 cases in high plasma viral DNA copies group and 37 cases in low copies group. More cases presented with elevated transaminases in the high plasma viral DNA copies group than those in the low group (75.7% (28/37) vs. 56.0%(116/207), χ2=5.00, P=0.025).Both the positive rate of EBV DNA in peripheral blood leukocytes (84.2% (266/316) vs. 44.7% (255/571), χ2=76.26, P<0.001) and the copies of EBV DNA (7.0×107 (1.3×107, 3.0×108) vs. 3.1×106 (1.6×106, 6.1×106) copies /L, Z=15.23, P<0.001) were higher than that of plasma. Conclusions: In immunocompetent pediatric cases diagnosed as EBV primary infection, cases with positive plasma EBV DNA were prone to have fever, hepatomegaly and (or) splenomegaly, and elevated transaminase than those with negative plasma viral DNA. The plasma EBV DNA usually turns negative within 28 days after initial diagnosis.Most cases with high viral load in plasma showed elevated aminotransferase.
Female
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Male
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Humans
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Child
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DNA, Viral
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Herpesvirus 4, Human
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Epstein-Barr Virus Infections
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Hepatomegaly
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Retrospective Studies
;
Splenomegaly
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Fever
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Transaminases
8.Effects of continuous goal-directed analgesia on fluid resuscitation of massive burn patients during shock.
Da Wei HAN ; Huan Na YANG ; Yan Cang LI ; Ying WEI ; She Min TIAN ; Cheng De XIA ; Xiao Liang LI
Chinese Journal of Burns 2022;38(1):38-44
Objective: To investigate the effects of continuous goal-directed analgesia on fluid resuscitation during shock stage in patients with massive burns, providing a basis for rational optimization of analgesia protocols in patients with burn shock. Methods: A retrospective case series study was conducted. One hundred and thirty-six patients with massive burns who met the inclusion criteria were admitted to Zhengzhou First People's Hospital from January 2015 to December 2020, and the patients were divided into continuous analgesia (CA) group (68 cases,with average age of 44 years old) and intermittent analgesia (IA) group (68 cases,with average age of 45 years old) according to whether sufentanil injection was continuously used for intravenous analgesia during the shock stage. The patients in the 2 groups were predominantly male. Before and at 72 h of treatment, the severity of disease and trauma pain of patients in the 2 groups were scored by the acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) and the visual analogue scale (VAS). Hematocrit, heart rate, mean arterial pressure (MAP), central venous pressure (CVP), oxygen saturation in central venous blood (ScvO2), rehydration coefficient, blood lactate value, hourly urine output, and the adverse reactions such as hypotension, nausea, vomiting, dizziness, skeletal muscle tonicity, respiratory depression, bradycardia, pruritus, and drug addiction of patients in the 2 groups during the treatment were recorded at the 1st, 2nd, and 3rd 24 h post-injury. Data were statistically analyzed with analysis of variance for repeated measurement, paired or independent sample t test, Bonferroni correction,chi-square test and Mann-Whitney U test. Results: Before treatment, APACHE Ⅱ and VAS scores of patients in the 2 groups were close (with t values of -0.67 and 0.32, respectively, P>0.05); At 72 h of treatment, APACHE Ⅱ and VAS scores of patients in CA group were 8.5±2.2 and 2.5±1.6, both of which were significantly lower than (15.2±3.0) and (7.9±2.0) of patients in IA group, respectively (with t values of -14.94 and -17.46, respectively, P<0.01). Compared with the pre-treatment period, the APACHE Ⅱ and VAS scores of patients in IA group decreased significantly at 72 h of treatment (with t values of 11.35 and 30.59, respectively, P<0.01); the changes in APACHE Ⅱ and VAS scores of patients at 72 h of treatment in comparison with those of patients before treatment in CA group were all similar to those of patients in IA group (with t values of 4.00 and 4.82, respectively, P<0.01). Compared with those of patients in IA group, there were no significant changes in CVP, hematocrit, heart rate, ScvO2, and MAP of patients in CA group at all three 24 h post-injury (with t values of <0.01, 0.12, 2.10, 1.55, 0.03; 0.13, 0.22, <0.01, 0.17, 0.49; 0.63, 0.06, 0.04, 2.79, and 2.33, respectively, P>0.05). Compared with those of patients in IA group at the 1st 24 h post-injury, CVP, ScvO2 and MAP of patients were significantly higher at the 2nd and 3rd 24 h post-injury (with t values of -10.10, -9.31, -8.89; -10.81, -4.65, and -9.43, respectively, P<0.01), and the heart rate of patients was significantly lower at the 2nd and 3rd 24 h post-injury (with t values of 7.53 and 7.78, respectively, P<0.01), and the hematocrit of patients decreased significantly only at the 3rd 24 h post-injury (t=15.55, P<0.01); the changes of CVP, ScvO2, MAP and heart rate of patients at the 2nd and the 3rd 24 h post-injury, and HCT of patients at the 3rd 24 h post-injury, in comparison with those of patients at the 1st 24 h post-injury in CA group were similar to those of patients in IA group (with t values of -12.25, -10.24, -8.99, 9.42, -8.83, -7.53, -11.57, 10.44, and 12.91, respectively, P<0.01). Compared with those of patients in IA group, the rehydration coefficient of patients in CA group was significantly higher only at the 3rd 24 h post-injury (t=5.60, P<0.05), blood lactate value of patients in CA group was significantly lower at the 1st and 2nd 24 h post-injury (with t values of 4.32 and 14.52, respectively, P<0.05 or P<0.01), the hourly urine output of patients in CA group increased significantly at the 1st, 2nd, and 3rd 24 h post-injury (with t values of 24.65, 13.12, and 5.63, respectively, P<0.05 or P<0.01). Compared with the those of patients at the 1st 24 h post-injury, the rehydration coefficient of patients in IA group decreased significantly at the 2nd and the 3rd 24 h post-injury (with t values of 33.98 and 36.91, respectively, P<0.01), the blood lactate values of patients in IA group decreased significantly at the 2nd and the 3rd 24 h post-injury (with t values of 8.20 and 11.68, respectively, P<0.01), and the hourly urine output of patients in IA group was significantly increased at the 2nd and the 3rd 24 h post-injury (with t values of -3.52 and -5.92, respectively, P<0.01); the changes of rehydration coefficients and blood lactate values of patients at the 2nd and the 3rd 24 h post-injury in comparison with those of patients at the 1st 24 h post-injury in CA group were similar to those of patients in IA group (with t values of 35.64, 33.64, 9.86, and 12.56, respectively, P<0.01), but hourly urine output of patients in CA group increased significantly only at the 3rd 24 h compared with that of patients at the 1st 24 h post-injury (t=-3.07, P<0.01). Adverse reactions such as hypotension, nausea, vomiting, dizziness, bradycardia, and pruritus occurred rarely in patients of the 2 groups, and none of the patients had skeletal muscle tonicity, respiratory depression, or drug addiction. The incidence of adverse reactions of patients in CA group was similar to that in IA group (χ2=0.08, P>0.05). Conclusions: Continuous goal-directed analgesia can effectively relieve pain and improve vital signs of patients with large burns. Meanwhile it has little impact on volume load, which can assist in correcting ischemia and hypoxia during the shock period and help patients get through the shock period smoothly.
Adult
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Analgesia
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Burns/therapy*
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Fluid Therapy
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Goals
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Humans
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Male
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Middle Aged
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Pain
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Resuscitation
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Retrospective Studies
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Shock/therapy*
9.Effect of Wenyang, Jieyu, and Wenyang Jieyu Prescriptions on Hippocampal Microglia of Mice with Depression-Like Behavior Induced by Secondary LPS Exposure
Zi-han GONG ; Jing-jing GAO ; Kai-jie SHE ; Huan-run ZHANG ; Yang ZUO ; Jing-wen LIANG ; Yuan LIANG ; Guang-xin YUE
Chinese Journal of Experimental Traditional Medical Formulae 2021;27(21):55-62
Objective:To observe the activation of microglia and the expression of inflammatory factors in hippocampus of mice with depression-like behavior after mother-infant separation (MS) combined with lipopolysaccharide (LPS) stress, and to explore the possible anti-depression mechanisms of Wenyang (WY), Jieyu (JY), and Wenyang Jieyu (WYJY) prescriptions from the perspective of warming Yang and relieving depression. Method:Seventy offspring mice were randomly divided into a normal group (
10.Changes of Microglia in Hippocampus of Mice Induced by Maternal Separation with Restraint Stress and Regulatory Effect of Wenyang Jieyu Prescription
Kai-jie SHE ; Jing-jing GAO ; Zi-han GONG ; Huan-run ZHANG ; Yang ZUO ; Jing-wen YANG ; Guang-xin YUE ; Yuan LIANG
Chinese Journal of Experimental Traditional Medical Formulae 2021;27(18):49-57
Objective:To observe the activation of microglia in hippocampus of depressed and anxious mice induced by maternal separation with acute restraint stress and the expression of interleukin-1

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