1.Effect of Scutellarin on the Expression and Activity of P-gp in Caco-2 Cells
Rui LIU ; Jing JIN ; Xiaoli GOU ; Ziyi ZHOU ; Yefeng CAI ; Min HUANG
China Pharmacist 2015;(4):529-532
Objective:To investigate the effect of scutellarin on P-gp protein expression and activity in Caco-2 cells. Methods:Scutellarin(25,50 and 100 μmol·L-1 )was incubated with Caco-2 cells respectively for 24 h,48 h and 72 h. The expression of P-gp was determined by western blot assay and the activity of P-gp was determined by Rhodamine-123 assay. Results:P-gp protein ex-pression levels were significantly increased by scutelarin. After the incubation for 24 h with scutellarin,P-gp protein expression was up-regulated 2. 34-,2. 65-and 2. 00-fold in Caco-2 cells. After the incubation with scutellarin for 48 h,P-gp protein expression was up-regulated 2. 70-,4. 66-and 3. 13-fold. After the incubation with scutellarin for 72 h,P-gp protein expression was up-regulated 2. 82-, 2. 62-and 1. 84-fold. The intracellular accumulation of rhodamine-123 was significantly decreased by scutellarin,indicating that the ef-flux transport activity of P-gp was increased by scutellarin in Caco-2 cells. Conclusion:Scutellarin can significantly up-regulate P-gp protein expression and increase the efflux transport activity of P-gp in Caco-2 cells.
2.Establishment of cell models for PXR, FXR and LXRαagonists high-throughput screening based on reporter gene assay
Jialang ZHUANG ; Hang ZENG ; Guoping ZHONG ; Jing JIN ; Xiaoli GOU ; Huichang BI ; Min HUANG
Chinese Pharmacological Bulletin 2015;(2):289-293
Aim To develop an in vitro high throughput drug screening system based on reporter gene assay for identification of novel compounds with PXR, FXR and LXRα agonist activity. Methods The expressions of exogenous PXR, FXR and LXRαgene in HEK293, HepG2 and LS174T cells were examined by Real-Time quantity PCR. pSG5-hPXR and pGL3-XREM-CYP3A4, pEGFP-N3-hFXR and EcRE-TK-Luc, pCMX-FLAG-hLXRα and pGL3-XREM-CYP3A4 were cotransfected into cells and the optimal ratio of three plasmids was determined. The dose-response relationship between the positive drug and the fold induction was determined. The specificity of the model was ex-amined, and the repeatability was also determined by Z′ value. Results ① The PXR, FXR and LXRα mRNA expression in HEK293 cell is low among three different cells. ②reporter gene vector and expression plasmid ratio of 1∶ 1, 2∶ 1 and 2∶ 1 were proved to be suitable for highest relative luciferase activity for PXR, FXR or LXRα agonist screening model. ③ The relative luciferase activity was induced by Rif, CDCA or T0901317 in a dose-dependent manner. ④Only Rif, CDCA or T0901317 could significantly increase the relative luciferase activity in PXR,FXR or LXRα agonist screening model, no effect of other nuclear re-ceptors agonist was observed, and the values of Z′-factor for PXR, FXR and LXRαagonist screening model were 0. 58, 0. 66 and 0. 63, respectively. Conclusion An in vitro PXR, FXR and LXRα agonist high-throughput screening models are devel-oped with acceptable specificity and repeatability, and the mod-els can be used to screen PXR, FXR and LXRα agonist.
3.Research on detection method and clinical countermeasure of clopidogrel-resisted metabolism-related genes after interventional operation of coronary heart disease (CHD)
Jun HE ; Yanhong ZHANG ; Zhiping GOU ; Yan WANG ; Xiaoli SUN ; Xiaohua LI
Journal of Chinese Physician 2015;17(7):1019-1023
Objective Study on cytochrome P450 (CYP) 2C19 gene single nucleotide polymorphism and the clinical prognosis of coronary heart disease (CHD) patients with percutaneous coronary intervention (PCI) after long-term use of clopidogrel.Methods A total of 150 cases of CHD patients was chosen prospectively between January 2013 and June 2014 who were hospitalized and PCI.All patients accepted dual antiplatelet therapy.Platelet aggregation rate and platelet aggregation inhibition rate were detected before taking the medicine and after PCI,which were used to classify the patients into clopidogrel-resistance groups (CR) and non-clopidogrel-resistance group (NCR).CYP2C19 gene single nucleotide polymorphism type was determined.The patients in CR group accepted clinical intervention countermeasures and NCR group was used as control,and postoperative recurrence angina and bleeding at the one year of operation were observed.Results About 24.67% of patients with CHD with clopidogrel treatment,platelet aggregation rate cannot recover to normal.The correlation analysis showed CYP2C19* 2 carriers had a significantly higher platelet aggregation rate.Comprehensive analysis found that CYP2C19 * 2,long-term smoking,increased platelet count,and diabetes were the independent risk factors that platelet aggregation rate cannot recover to normal.No significant differences were found in primary end point,secondary end points,and bleeding events between CR group after clinical intervention countermeasures and NCR group.Conclusions CYP2C19 * 2,long-term smoking,increased platelet count,and diabetes are the independent risk factors that platelet aggregation rate cannot be developed to standard.Patients with clopidogrel resistance,the clinical intervention countermeasures to strengthen antiplatelet therapy can improve high platelet reactivity after PCI in CHD patients,and does not increase the risk of bleeding.
4.Research progress in diagnosis, treatment and rehabilitation of deltoid ligament injury of ankle joint
Xuting BIAN ; Xiaoli GOU ; Xuan ZHANG ; Kanglai TANG ; Xu TAO
Chinese Journal of Trauma 2022;38(9):840-848
As the main stabilizing structure of the medial ankle joint, deltoid ligament plays a role in counteracting excessive eversion of the hindfoot and external rotation of the talus during ankle movement so as to maintain the biomechanical stability of ankle joint. Although the incidence of deltoid ligament injury is low, improper diagnosis and treatment can affect the path of talus motion and eventually lead to chronic medial instability or traumatic arthritis of the ankle joint, seriously affecting the normal life and motor function of the patients. The diagnosis of deltoid ligament injury needs to be based on the characteristics of the injury, physical examination and imaging, among which X-ray, MRI and ultrasonography are most frequently used. There are various methods to treat deltoid ligament injury according to the type of injury, and thus the choice of treatment has been a hot topic in the field of foot and ankle surgery. The choice of non-surgical or surgical treatment for acute deltoid ligament injury remains controversial. For the treatment of chronic deltoid ligament injury, there is no consensus on direct repair or deltoid ligament reconstruction. In addition, the choice of autologous or allograft tendon or wire anchors for deltoid ligament reconstruction is also disputed. The rehabilitation of deltoid ligament injury is crucial to the early restoration of motor function of the ankle joint, but the related guidelines or consensus are scarce. In order to fully understand the characteristics of deltoid ligament injury, make accurate diagnosis and formulate reasonable treatment and rehabilitation programs, the authors review the research progress in deltoid ligament injury from aspects of anatomical characteristics, biomechanical mechanism of injury, diagnosis, treatment and postoperative functional rehabilitation, hoping to provide a reference for the clinical diagnosis and treatment of deltoid ligament injury.
5.The value of combined detection of serum neurogranin and hypoxia-inducible factor-1α on the prognosis in patients with severe craniocerebral trauma
Xiaoli HUANG ; Xin GOU ; Kaixuan FENG ; Jiebing LING ; Bin MA ; Hao ZHOU ; Xiangyang LUO
Chinese Journal of Postgraduates of Medicine 2021;44(7):621-626
Objective:To investigate the value of combined detection of serum neurogranin (NG) and hypoxia-inducible factor-1α (HIF-1α) in patients with severe craniocerebral trauma.Methods:Ninety-seven patients with severe craniocerebral trauma from June 2018 to March 2020 in Jinshan Branch of Shanghai Sixth People′s Hospital were selected. According to the Glasgow outcome score (GOS), 97 patients were divided into good prognosis group (GOS>3 scores, 46 cases) and poor prognosis group (GOS ≤ 3 scores, 51 cases). The NG, HIF-1α, Glasgow coma score (GCS), acute physiology and chronic health status score Ⅱ (APACHE Ⅱ) were compared between 2 groups. The independent risk factors of prognosis in patients with severe craniocerebral trauma were analyzed by multivariate Logistic regression analysis. The diagnostic efficacy of NG and HIF-1α on poor prognosis in patients with severe craniocerebral trauma was analyzed by receiver operating characteristic (ROC) curve. The correlation between serum NG, HIF-1α and APACHE Ⅱ in patients with severe craniocerebral trauma was analyzed by Pearson analysis.Results:The GCS in good prognosis group was significantly higher than that in poor prognosis group: (6.50 ± 1.74) scores vs. (4.76 ± 0.78) scores, the NG, HIF-1α and APACHE Ⅱwere significantly lower than those in poor prognosis group: (696.98 ± 158.96) ng/L vs. (875.92 ± 188.52) ng/L, (34.72 ± 13.98) μg/L vs. (51.29 ± 14.17) μg/L and (15.69 ± 3.45) scores vs. (22.58 ± 6.45) scores, and there were statistical differences ( P<0.01). Multivariate Logistic regression analysis result showed that the NG, HIF-1α, APACHEⅡ, GCS and type of craniocerebral trauma were independent risk factors on the prognosis in patients with severe craniocerebral trauma ( P<0.05 or<0.01). ROC curve analysis result showed that the AUC of NG and HIF-1αNG and HIF-1α combined detection to assess the poor prognosis in patients with severe craniocerebral trauma was significantly higher than NG and HIF-1α alone detection (0.873 vs. 0.772 and 0.821, Z = 2.276 and 1.949, P<0.05). Pearson correlation analysis result showed that APACHE Ⅱ was positive correlation with serum NG and HIF-1α in severe craniocerebral trauma patients with poor or good prognosis ( r = 0.852 and 0.889, P<0.01; r = 0.717 and 0.851, P<0.01). Conclusions:The combined detection of serum NG and HIF-1α can be used as an evaluation index for the prognosis in patients with severe craniocerebral trauma, which helps to determine the severity of craniocerebral trauma and has great value for clinical diagnosis and treatment.
6.Effect of caffeine on hyperoxic lung injury of premature rats and its relationship with p38 motigen ̄acti ̄vated protein kinase signal pathway
Yanan SONG ; Yi YANG ; Enjin GOU ; Huiling SONG ; Qing LI ; Huan WANG ; Xiang ZHANG ; Kaifeng WU ; Xiaoli LIU ; Xinghui ZHENG ; Bo HUANG
Chinese Pediatric Emergency Medicine 2019;26(6):454-460
Objective To study the effect of caffeine on hyperoxic lung injury of premature rats and its relationship with p38 motigen-activated protein kinase( MAPK) signal pathway. Methods Sixty Wistar premature rats were divided into 4 groups(n=15) according to the random number table:air + normal sa-line group(A+N group),air + caffeine group( A+C group),hyperoxia + normal saline group( H+N group),and hyperoxia + caffeine group(H+C group). Among them,H+N group and H+C group were continually exposed to hyperoxia ( oxygen concentration was 60% ~70%). For A + C group and H + C group,the premature rats were injected with caffeine of 29 mg/(kg·d) into their peritoneal cavities every day after birth. For A+N group and H+N group,the premature rats were injected with normal saline of the same volume into their peritoneal cavities. In each group,the lung tissues of 5 premature rats were randomly select-ed on the third, seventh and fourteenth day respectively. The pathological changes of lung tissue, radiated alveolar count(RAC) and collagen content in lung tissue were observed under a light microscope. The wet/dry ratio ( W/D) was measured. Two-step immunohistochemistry was used to detect the distribution of p38MAPK in lung tissue. The content of phosphorylated p38MAPK( p-p38MAPK) protein was detected by western blot. Results Compared with the air groups,the lung tissues of premature rats in high oxygen expo-sure groups showed different degree of inflammatory changes on the third,seventh,and fourteenth day. The changes were more obvious with the prolonged exposure to hyperoxia. Pulmonary fibrosis was visible on the fourteenth day,which was improved after caffeine intervention. The RAC value of premature rats in hyperoxia exposure groups was significantly lower than that in air-exposure groups(P<0. 05),and the W/D ratio and collagen content in lung tissue increased significantly (P<0. 05),which were improved after caffeine inter-vention(P<0. 05). The results of two-step immunohistochemistry showed that the number of p-p38MAPK positive cells in the lung tissue of premature rats in hyperoxia exposure groups increased and widely distribu-ted, but decreased after caffeine intervention. The results of western blot showed that the content of p-p38MAPK protein in lung tissue of premature rats in hyperoxia exposure groups was significantly higher than that of air groups(P<0. 05),but it decreased after caffeine intervention(P<0. 05). Conclusion Hy-peroxia can promote the formation of pulmonary fibrosis by activating p38MAPK signal pathway. Caffeine can interdict the expression of p38MAPK to alleviate the fibrosis of lung tissue exposed to hyperoxia and thus protects the lung tissue.
7.Biplane osteotomy to treat malunion of Stephens-Sanders type Ⅱ calcaneal fracture
Zhenyu WANG ; Fangcheng YANG ; Mei CHENG ; Guo ZHENG ; Qiankun NI ; Yuanqiang LI ; Xiaoli GOU ; Kanglai TANG ; Xu TAO
Chinese Journal of Orthopaedic Trauma 2023;25(10):833-839
Objective:To evaluate the clinical efficacy of biplane osteotomy in the treatment of malunion of Stephens-Sanders type Ⅱ calcaneal fracture.Methods:A retrospective study was conducted to analyze the clinical data of 31 patients who had been treated by biplane osteotomy at Sports Medicine Center, The First Affiliated Hospital of Army Medical University for malunion of Stephens-Sanders type Ⅱ calcaneal fracture from January 2019 to January 2022. There were 21 males and 10 females, with an age of (41.4±13.9) years and a duration from injury to diagnosis of (12.8±8.9) months. Functional and image scores were compared before surgery, 6 months after surgery, and at the last follow-up. Functional scores included the visual analogue scale (VAS) score, the American Orthopedic Foot and Ankle Society (AOFAS) score, and the pain interference (PI) and physical function (PF) indices in the Patient-Reported Outcomes Measurement Information System (PROMIS). Image scores included the Gissane angle, B?hler's angle, calcaneal pitch angle, length of the calcaneus, absolute foot height, and axial calcaneal width as measured on X-rays.Results:The operation time was (106.6±29.9) minutes for this cohort. All the 31 patients were followed up for (18.4±5.8) months. At 6 months after surgery and the last follow-up, the VAS scores [3 (2, 3), 2 (1, 3)], AOFAS scores [83 (76, 87), 85 (83, 87)], PI scores [(57±9), (48±6)], PF scores [53 (39, 61), 56 (54, 66)], Gissane angles (109.6°±14.1°, 109.3°±14.9°), B?hler angles (26.5°±11.6°, 26.9°±11.8°), calcaneal pitch angles [19.1° (14.5°, 23.9°), 19.9° (14.5°, 23.9°)], absolute foot heights [(76.5±9.6) mm, (76.0±9.9) mm], and axial calcaneal widths [(38.5±4.1) mm, (38.3±4.1) mm] were all significantly improved compared to the preoperative values [5 (4, 6), 62 (56, 67), (62±6), 47 (38, 51), 126.8°±13.1°, 11.8°±10.9°, 13.8° (8.2°, 18.7°), (71.0±9.1) mm, (42.8±5.5) mm] (all P<0.05). However, there was no statistically significant difference in the length of the calcaneus among pre-surgery, 6 months after surgery, and the last follow-up ( P>0.05). Conclusion:Biplane osteotomy is a surgical technique that demonstrates good clinical efficacy in the treatment of malunion of Stephens-Sanders type Ⅱ calcaneal fracture so that it should be promoted in clinic.
8. LncRNA ANRIL target miR-195 experimental study of radiation sensitivity of HCT116 cells and nude mouse transplant tumors
Xiaoyan CHEN ; Chenbin WU ; Xin TIAN ; Xiaoli GOU ; Yingqiang WU ; Kui ZHAO ; Rui XIE
Chinese Journal of Radiation Oncology 2019;28(11):858-861
Objective:
To investigate the effect and mechanism of LncRNA ANRIL on the radiosensitivity of HCT116 cells line and nude mouse transplant tumors.
Methods:
The expression of LncRNA ANRIL in colorectal cancer cells was detected by qPCR. The negative control siRNA, ANRIL siRNA, miR-NC mimic, miR-195 mimic, miR-NC inhibitor and miR-195 inhibitor were transfected into HCT116 cells, and marked as negative control group, silencing ANRIL group, overexpressing miR-NC group, overexpressing miR-195 group, inhibiting miR-NC group and inhibiting miR-195 group, and the HCT116 cells without any treatment were marked as the blank control group. The clone formation assay was used to detect radiosensitivity of colorectal cancer cells, flow cytometry was used to detect apoptosis. The web site, StarBase, was used to predict the downstream miRNAs of ANRIL and dual luciferase reporter gene assay was used to further verify. Subcutaneous tumor transplantation assay was used to detect the effect of ANRIL on the growth of colorectal cancer cells after irradiation.
Results:
After irradiation with 2, 4, 6 and 8 Gy, the cell survival fraction of silencing ANRIL group was significantly decreased when compared with that of negative control group (