1.Effect of Chaibei Zhixian Decoction on intestinal flora and Th17/Treg cell immune balance in epileptic rats by regulating mTOR/HIF-1α pathway
Rui ZHAO ; Ling GAO ; Youfeng WANG ; Xian REN ; Fangjing SU ; Xiaoyan HAN
Chinese Journal of Immunology 2025;41(1):129-135
Objective:To explore the effect of Chaibei Zhixian Decoction on intestinal flora and T helper 17/regulatory T cell(Th17/Treg)immune balance in epileptic rats by regulating mammalian target of rapamycin/hypoxia-inducible factor-1α(mTOR/HIF-1α)pathway.Methods:Sixty rats were randomly divided into Sham group,Model group,low-dose Chaibei Zhixian Decoction group(CBZXD-Low)and high-dose Chaibei Zhixian Decoction group(CBZXD-High),with 15 rats in each group.Lateral ventricular injec-tion 1 μl kainic acid(1.5 μg/μl)was used to prepare epileptic rat model,Sham group was injected with an equal amount of physiological saline.CBZXD-Low group and CBZXD-High group were given Chaibei Zhixian Decoction by gavage daily(8.48 g/kg,16.96 g/kg),while Sham group and model group were given an equal amount of physiological saline by gavage.After continuous gavage for 4 weeks,HE staining and Nissl staining were used to detect pathological damage in hippocampal tissue;RT-qPCR was used to detect Th17/Treg related marker mRNA levels in spleen tissue and mTOR,HIF-1α mRNA levels in hippocampus tissue;flow cytometry was used to de-tect Th17/Treg cell ratio in spleen tissue;16S rRNA sequencing of feces in colon segment was performed;Western blot was used to de-tect mTOR and HIF-1α protein levels in hippocampal tissue.Results:Compared with Sham group,Model group rats suffered from spontaneous epilepsy,the arrangement of hippocampal neurons was disordered and severely lacking,intracellular Nissl bodies was de-creased,RORγt,IL-17A mRNA levels in spleen tissue were increased,Foxp3,CTLA-4 and GITR mRNA levels were decreased,the proportion of Th17 cells and Th17/Treg were increased,the proportion of Treg cell was decreased,intestinal flora α/β diversity were decreased,the proportion of harmful bacteria such as Cronobacter and Heliobacillus were increased,the proportion of beneficial bacte-ria such as Bacteroides,Lactobacillus,Prevotella and Akkermansia were decreased;mTOR and HIF-1α mRNA levels,p-mTOR/mTOR,HIF-1α protein levelsin hippocampal tissue were increased(P<0.05);compared with Model group,the seizures of rats in CBZXD-Low group and CBZXD-High group were significantly reduced,and the damage to hippocampal neurons was alleviated,RORγt,IL-17A mRNA levels in spleen tissue were decreased,Foxp3,CTLA-4 and GITR mRNA levels were increased,the propor-tion of Th17 cells and Th17/Treg were decreased,the proportion of Treg cell was increased,intestinal flora α/β diversity were in-creased,the proportion of harmful bacteria such as Cronobacter and Heliobacillus were decreased,the proportion of beneficial bacte-ria,such as Bacteroides,Lactobacillus,Prevotella,and Akkermansia were increased;mTOR and HIF-1α mRNA levels,p-mTOR/mTOR,HIF-1α protein levels in hippocampal tissue were decreased(P<0.05);and it showed a dose-dependent effect of Chaibei Zhixian Decoction(P<0.05).Conclusion:Chaibei Zhixian Decoction can restore intestinal flora and Th17/Treg cell immune balance in epileptic rats,it possibly plays a role by regulating mTOR/HIF-1α pathway.
2.Effect of Chaibei Zhixian Decoction on intestinal flora and Th17/Treg cell immune balance in epileptic rats by regulating mTOR/HIF-1α pathway
Rui ZHAO ; Ling GAO ; Youfeng WANG ; Xian REN ; Fangjing SU ; Xiaoyan HAN
Chinese Journal of Immunology 2025;41(1):129-135
Objective:To explore the effect of Chaibei Zhixian Decoction on intestinal flora and T helper 17/regulatory T cell(Th17/Treg)immune balance in epileptic rats by regulating mammalian target of rapamycin/hypoxia-inducible factor-1α(mTOR/HIF-1α)pathway.Methods:Sixty rats were randomly divided into Sham group,Model group,low-dose Chaibei Zhixian Decoction group(CBZXD-Low)and high-dose Chaibei Zhixian Decoction group(CBZXD-High),with 15 rats in each group.Lateral ventricular injec-tion 1 μl kainic acid(1.5 μg/μl)was used to prepare epileptic rat model,Sham group was injected with an equal amount of physiological saline.CBZXD-Low group and CBZXD-High group were given Chaibei Zhixian Decoction by gavage daily(8.48 g/kg,16.96 g/kg),while Sham group and model group were given an equal amount of physiological saline by gavage.After continuous gavage for 4 weeks,HE staining and Nissl staining were used to detect pathological damage in hippocampal tissue;RT-qPCR was used to detect Th17/Treg related marker mRNA levels in spleen tissue and mTOR,HIF-1α mRNA levels in hippocampus tissue;flow cytometry was used to de-tect Th17/Treg cell ratio in spleen tissue;16S rRNA sequencing of feces in colon segment was performed;Western blot was used to de-tect mTOR and HIF-1α protein levels in hippocampal tissue.Results:Compared with Sham group,Model group rats suffered from spontaneous epilepsy,the arrangement of hippocampal neurons was disordered and severely lacking,intracellular Nissl bodies was de-creased,RORγt,IL-17A mRNA levels in spleen tissue were increased,Foxp3,CTLA-4 and GITR mRNA levels were decreased,the proportion of Th17 cells and Th17/Treg were increased,the proportion of Treg cell was decreased,intestinal flora α/β diversity were decreased,the proportion of harmful bacteria such as Cronobacter and Heliobacillus were increased,the proportion of beneficial bacte-ria such as Bacteroides,Lactobacillus,Prevotella and Akkermansia were decreased;mTOR and HIF-1α mRNA levels,p-mTOR/mTOR,HIF-1α protein levelsin hippocampal tissue were increased(P<0.05);compared with Model group,the seizures of rats in CBZXD-Low group and CBZXD-High group were significantly reduced,and the damage to hippocampal neurons was alleviated,RORγt,IL-17A mRNA levels in spleen tissue were decreased,Foxp3,CTLA-4 and GITR mRNA levels were increased,the propor-tion of Th17 cells and Th17/Treg were decreased,the proportion of Treg cell was increased,intestinal flora α/β diversity were in-creased,the proportion of harmful bacteria such as Cronobacter and Heliobacillus were decreased,the proportion of beneficial bacte-ria,such as Bacteroides,Lactobacillus,Prevotella,and Akkermansia were increased;mTOR and HIF-1α mRNA levels,p-mTOR/mTOR,HIF-1α protein levels in hippocampal tissue were decreased(P<0.05);and it showed a dose-dependent effect of Chaibei Zhixian Decoction(P<0.05).Conclusion:Chaibei Zhixian Decoction can restore intestinal flora and Th17/Treg cell immune balance in epileptic rats,it possibly plays a role by regulating mTOR/HIF-1α pathway.
3.Status of surgical treatment of aortic diseases: experience of multi-center aortic registry in Jiangsu Province
Cunhua SU ; Fangjing ZHENG ; Xin CHEN ; Fuhua HUANG
Chinese Journal of Thoracic and Cardiovascular Surgery 2022;38(12):716-720
Objective:To analyze the status of treatment of aortic diseases in nine centers included in Jiangsu Aortic Registry study.Methods:Medical records of 1 105 patients who underwent surgery for aortic diseases in nine cardiovascular centers in Jiangsu province from January 1, 2019 to December 31, 2020 were retrospectively collected. According to the types of diseases, they are divided into three categories: Category Ⅰ diseases: aortic diseases requiring only simple open surgery; Category Ⅱ: aortic disease requiring complex open surgery; Category Ⅲ disease: aortic disease requiring interventional or hybrid surgery. The diagnosis and treatment of the three diseases were compared by statistical analysis.Results:Patients aged 50-59 years and 60-69 years were the most in the whole group, accounting for 291(26.3%) and 319 (28.9%), respectively. That is, the age group between 50-69 years was the main age for the incidence of aortic disease, accounting for 55.2% (610/1 105) of the total incidence. The ratio of male to female was 803∶302, and male was 72.7% (803/1 105). The perioperative mortality of patients with type Ⅰ and Ⅲ diseases was lower [1.6%(6/385) and 1.0% (4/330), respectively]. The perioperative mortality of patients with type Ⅱ diseases was significantly increased (15.5% (51/330)) due to their dangerous onset and complicated operation. The highest postoperative complications were infection 11.3% (124/1 105), acute kidney injury 8.6% (95/1 105), and neurological complications 7.0% (77/1 105), respectively.Conclusion:Individualized treatment according to the lesion site, lesion type and scope of involvement of aortic disease can achieve a satisfactory prognosis.
4.Influence of prior percutaneous coronary intervention on outcome of coronary artery bypass grafting: A multi-center clinical study
Hongwei JIANG ; Hang ZHANG ; Wen CHEN ; Fangjing ZHENG ; Yongfeng SHAO ; Yongxiang QIAN ; Guoxiang WANG ; Mingqiu LI ; Qingsheng YOU ; Zhiyong LIU ; Yong WANG ; Zhenya SHEN ; Wei LI ; Demin LI ; Su HUANG ; Chongjun ZHONG ; Rui WANG ; Xin CHEN
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2022;29(11):1436-1441
Objective To investigate the influence of prior percutaneous coronary intervention (PCI) on the outcome of coronary artery bypass grafting (CABG). Methods Clinical data of 5 216 patients from Jiangsu Province CABG registry who underwent primary isolated CABG from 2016 to 2019 were retrospectively analyzed. Patients were divided into a PCI group (n=673) and a non-PCI group (n=4 543) according to whether they had received PCI treatment. The PCI group included 491 males and 182 females, aged 62.6±8.2 years, and the non-PCI group included 3 335 males and 1 208 females, aged 63.7±8.7 years. Multivariable logistic regression and propensity score matching (PSM) were used to compare 30-day mortality, incidence of major complications and 1-year follow-up outcomes between the two groups. Results Both in original cohort and matched cohort, there was no statistical difference in the 30-day mortality [14 (2.1%) vs. 77 (1.7%), P=0.579; 14 (2.1%) vs. 11 (1.6%), P=0.686], or the incidence of major complications (myocardial infarction, stroke, mechanical ventilation≥24 h, dialysis for new-onset renal failure, deep sternal wound infection and atrial fibrillation) (all P>0.05). The rate of reoperation for bleeding in the PCI group was higher than that in the non-PCI group [19 (2.8%) vs. 67 (1.5%), P=0.016; 19 (2.8%) vs. 7 (1.0%), P=0.029]. Both in original cohort and matched cohort, there was no statistical difference in 1-year survival rate between the two groups [613 (93.1%) vs. 4 225 (94.6%), P=0.119; 613 (93.1%) vs. 630 (95.2%), P=0.124], while the re-admission rate in the PCI group was significantly higher than that in the non-PCI group [32 (4.9%) vs. 113 (2.5%), P=0.001; 32 (4.9%) vs. 17 (2.6%), P=0.040]. Conclusion This study shows that a history of PCI treatment does not significantly increase the perioperative mortality and major complications of CABG, but increases the rate of cardiogenic re-admission 1 year postoperatively.
5.Effects of andrographolide on invasion and apoptosis of ovarian cancer SKOV-3 cells
Fangjing SU ; Bin ZHANG ; Linyan TIAN
Chinese Journal of Pathophysiology 2017;33(7):1328-1331
AIM: To investigate the effects of andrographolide on the invasion and apoptosis of ovarian cancer cell line SKOV-3, and to explore the possible mechanisms.METHODS: SKOV-3 cells were treated with different concentrations (0, 5, 10, 20 or 40 μmol/L) of andrographolide for different time (12, 24, 36 or 48 h), and then the cell viability was determined by CCK-8 assay.The cell invasion ability was analyzed by Transwell assay and cell apoptosis was detected by TUNEL staining.The protein levels of p-PI3K, p-Akt and p-mTOR were examined by Western blot.RESULTS: The results of CCK-8 assay revealed that andrographolide inhibited the growth of SKOV-3 cells in a dose-and time-dependent manner.Treatment with andrographolide at 20 μmol/L for 36 h significantly decreased the invasion ability of SKOV-3 cells, while increased cell apoptosis.In addition, the protein levels of p-PI3K, p-Akt and p-mTOR were reduced after andrographolide treatment.CONCLUSION: Andrographolide inhibits the growth and invasion of ovarian cancer SKOV-3 cells by suppression of PI3K/Akt/mTOR signaling pathway.

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