1.Potassium channels in vascular smooth muscle and essential hypertension
Chinese Journal of Pathophysiology 1986;0(04):-
Essentialhypertension(EH)ischaracterizedbyanincreasedtotalperipheralresistance . Therearefourtypesofpotassiumchannelsinvascularsmoothmusclecells ,includingKca ,Kv ,Kir ,KATP , whichplayanimportantroleinregulatingthediameterofvascular .Thechangeofpotassiumchannelsmay havesomethingtodowiththepathogenesisofhypertension .Thisarticlereviewsthecharactersofpotassium channelsandtheirrolesinEH . [
2.Correlative cell signaling pathway for expression of heme oxygenase-1 induced by ginkgo biloba extract in rat vascular smooth muscle cells
Xiaoyu YANG ; Hongxia WANG ; Zongde WEI ; Weiyi HUANG
Chinese Journal of Pathophysiology 2000;0(12):-
AIM: To explore the effects of heme oxygenase-1(HO-1) protein expression induced by ginkgo biloba extract(EGB761) in rat vascular smooth muscle cells(RVSMC) and the correlative cell signaling pathway.METHODS: The RVSMC lines were revived.Serial passage to 6 generation was carried out and divided into different groups.The cells were treated respectively with vehicle,purely EGB761,EGB761 plus zinc protoporphyrin IX or other specific inhibitors of cell signaling pathway.Western blotting method was used to detect the expression of HO-1 in RVSMC.RESULTS: EGB761 induced HO-1 protein expression in a dose dependent manner.ZnPPⅨ and genitein significantly inhibited HO-1 protein expression induced by EGB761(0.10?0.01,0.07?0.01 vs 0.61?0.07,P0.05,respectively).CONCLUSION:(1) EGB761 significantly induces HO-1 protein expression in RVSMC,and the effect can be inhibited by a specific HO inhibitor ZnPPⅨ.(2) The HO-1 protein expression induced by EGB761 in RVSMC is mediated by tyrosine protein kinase pathway.
3.The auxiliary diagnosis value of enzyme-linked immunosorbent spot assay and flow cytometry in detection of IFN-γ in tuberculous pleurisy
Fengjiao DU ; Qiping GE ; Panjian WEI ; Hongyan JIA ; Shuxiang GU ; Zongde ZHANG
Chinese Journal of Laboratory Medicine 2011;34(7):617-622
Objective To explore the value of IFN-γ produced or secreted by CD+4 T Lymphocytes from pleural effusion mononuclear cells for the diagnosis of tuberculous pleurisy(plTB).Methods The PEMCs of 40 patients with tuberculous pleural effusion and 30 patients with malignancy pleural effusion were selected as the tuberculosis and disease control groups, then co-cultured with the early secretory antigenic target 6 (ESAT-6) and culture filtered protein 10 (CFP-10) fusion protein (E/C).The numbers of spot forming cells(SFC) secreting IFN-γ were enumerated by ELISpot and the ratios of cells producing IFN-γ were detected by flow cytometry and intracellular cytokine staining.Moreover, the two indicators were compared between tuberculosis and disease control groups to evaluate the 2 methods detecting IFN-γ in the diagnosis of plTB.Results After E/C stimulation, the numbers of SFC were 205(125-450)SFC/5×104 PEMC in tuberculosis group and 5(2-18)SFC/5×104 PEMC in disease control group by ELISpot.The difference between two groups was statistically significant (U= 20.00, P<0.01).The proportion of IFN-γ-secreting CD+4 T lymphocytes was 3.27% (1.81%-7.34%) in tuberculosis group and 0.12% (0.06%-0.46%) in control group detected by FCM. The difference between the two groups was statistically significant (U=45.00, P<0.01).The indicators of ELISpot in detection of IFN-γ which was secreted by PEMC after co-cultured with E/C were as follows: sensitivity 92.5% (37/40), specificity 80.0% (24/30), positive predictive value 0.86, negative predictive value 0.89, positive likelihood ratio 4.63, negative likelihood ratio 0.09 and accuracy 87.1%;and for FCM, they were 87.5% (35/40), 90.0% (27/30), 0.92, 0.84, 8.75 and 0.14, respectively and accuracy 88.6%.Conclusion After E/C stimulation, the assay for IFN-γ-secreting CD+4 T lymphocytes by FCM and ELISpot is highly sensitive and specific for diagnosis of plTB as an auxiliary method.
4.Observation on the functional characteristics of multidrug-resistant Mycobacterium tuberculosis-infected macrophage model
Xiali LYU ; Tingting LIN ; Jingtao GAO ; Hongyan JIA ; Chuanzhi ZHU ; Zihui LI ; Jing DONG ; Qi SUN ; Wei SHU ; Liping PAN ; Zongde ZHANG ; Qi LI
Chinese Journal of Microbiology and Immunology 2021;41(10):741-748
Objective:To observe the characteristics of the phagocytosis and bactericidal function of multidrug-resistant Mycobacterium tuberculosis(MDR- Mtb)-infected macrophage model, and the changes of the immune response and metabolic function in the process of phagocytosis and bactericidal function, aiming to provide reference for studying the role and mechanism of macrophages in the occurrence and development of multidrug-resistant tuberculosis(MDR-TB). Methods:We established MDR- Mtb and H37Rv-infected macrophage models, and used the colony-forming unit (CFU), Magnetic Luminex ? Assay and Cholesterol Assay kit to observe the effects on phagocytosis and bactericidal function, the secretion of Th1(IL-12/23 p40, IL-27 and TNF-α) and Th2 cytokines (IL-6 and IL-10) and cholesterol metabolism. The data were analyzed by SPSS25.0 software. The data were expressed as Mean± SD and analyzed by t test or F test. P<0.05 was considered statistically significant. Results:(1) After MDR- Mtb-infected macrophages, the intracellular CFU gradually increased and reached the highest at 24 h, while the extracellular CFU gradually decreased and reached the lowest at 24 h. The intracellular CFU at 48 h was lower than that at 24 h, while the extracellular CFU was higher than that at 24 h ( P<0.05). Both intracellular and extracellular CFU at 48 h were close to those at 4 h ( P>0.05). The intracellular CFU was lower than the H37Rv group at 8-48 h, while the extracellular CFU was higher than the H37Rv group ( P<0.05). (2) The level of IL-12/23 p40, IL-27, TNF-α, IL-6 and IL-10 of MDR-TB group were higher than those of blank group ( P<0.05), but the level of TNF-α and IL-6 at 24 h and 48 h were higher than that at 4 h ( P<0.05). IL-12/23 p40 and TNF-α at 48 h and IL-6 at 24 h were lower than those of the H37Rv group, while IL-27 at 48 h was higher than that of the H37Rv group ( P<0.05). (3) The levels of cholesterol of MDR-TB group at 24 h and 48 h were lower than those of 4 h and blank group ( P<0.05), but the level of cholesterol was similar to the H37Rv group at any time ( P>0.05). (4) TNF-α reached the highest when the intracellular CFU reached the highest at 24 h, and IL-6 reached the highest when the intracellular CFU decreased at 48 h. With the decreasing of cholesterol expression, the intracellular CFU increased and then decreased. Conclusions:MDR- Mtb could induce the phagocytosis and bactericidal function of macrophages, increase the expression of Th1 and Th2 cytokines and promote the utilization and consumption of cholesterol, but this function was weaker than that of H37Rv strain.
5.The optimal reference genes for miRNA quantitative analysis in cerebrospinal fluid of patients with tuberculous meningitis
Hongyan JIA ; Mailing HUANG ; Fei LIU ; Jing DONG ; Qi SUN ; Rongrong WEI ; Aiying XING ; Boping DU ; Zongde ZHANG ; Liping PAN
Chinese Journal of Laboratory Medicine 2020;43(7):732-738
Objective:To explore the feasibility of U6 and Cel-miR-39 as reference genes for quantitative detection of microRNA (miRNA) in cerebrospinal fluid (CSF) of tuberculous meningitis (TBM), and validate the difference of miRNAs between tuberculous and viral meningitis (VM).Methods:The remaining CSF specimens after routine examination were collected in Beijing Chest Hospital of Capital Medical University. A total of 36 TBM and 34 VM patients were enrolled based on the information in the medical records. Total RNA were extracted from the CSF samples, and Taqman based real-time quantitative PCR (RT-CR) analysis were performed to determine the concentration of the miRNAs in CSF. GeNorm, NormFinder and Bestkeeper software were used for stability analysis of the two reference genes. 2 -ΔCt method was used to determine the relative gene expression. Accordance of repeated tests was analyzed by Pearson correlation test. Continuous variables were compared by the t-test. Results:Among the 70 samples, the average cycle threshold (Ct) value of U6 was 30.40±3.30, while the average Ct value of Cel-miR-39 was 21.49±0.70. The expression level of Cel-miR-39 was higher than that of U6. Correlation analysis showed good accordance of the repeated tests among the reference genes and target genes analysis in the randomly selected 10 samples ( r>0.931, P<0.001). Based on the analyses results of the three software, including GeNorm, NormFinder and Bestkeeper, Cel-miR-39 presented better stability in RT-PCR analysis and was more suitable as a reference gene for miRNA quantitative determination in CSF sample of TBM patients. The relative expression levels of the three target miRNAs were calculated using Cel-miR-39 as the reference gene, and miR-126-3p (1.13±0.41 vs 3.34±0.82, t=2.452, P=0.016), miR-130a-3p (0.56±0.10 vs 2.59±0.70, t=2.960, P=0.004) and miR-151a-3p (0.64±0.25 vs 2.11±0.33, t=3.536, P<0.001) were showed significant lower expression levels in CSF in TBM group than that in VM group. Conclusions:Cel-miR-39 can be used as a reference gene for quantitative detection of miRNAs in CSF of TBM patients. Significant differences were detected in expression level of miR-126-3p, miR-130a-3p and miR-151a-3p between TBM and VM group.