1.Role of PPARγ on γ-glutamylcysteine synthetase activity and expression in rats with chronic obstructive pulmonary disease
Jie LI ; Aiguo DAI ; Ruicheng HU ; Liming ZHU ; Meifang WANG
Chinese Journal of Pathophysiology 2010;26(2):302-308
AIM: To explore the effects of peroxisome proliferator-activated receptor γ (PPARγ) on the activity and expression of γ-glutamylcysteine synthetase (γ-GCS), and its role in rats with chronic obstructive pulmonary disease. METHODS: COPD model was established by the method of combining fumigation and lipopolysaccharide (LPS) intra-tracheal dripping. Meanwhile, some of the COPD rats were administered with rosiglitazone (RGZ), a PPARγ activator. The pulmonary function and the pathological changes were determined. ROS content and γ-GCS activity in lung tissues were detected. The levels of PPARγ, γ-GCS mRNA and protein expression in lung tissues were measured by immunohistochemistry, Western blotting, in situ hybridization (ISH) and reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: The pulmonary function (FEV_(0.3), FEV_(0.3)/FVC%, PEF) were significantly improved in RGZ group compared to COPD group. Under light microscope, lung pathological changes in COPD group conformed to pathological features of COPD. The pathological changes of lung tissue were obviously reduced in RGZ group compared to COPD group. In RGZ group, ROS content was obviously reduced and γ-GCS activity significantly increased compared to COPD group. Protein and mRNA expressions of PPARγ and γ-GCS in COPD group significantly higher than those in control group (all P<0.01), and those in RGZ group was markedly increased compared to COPD group (all P<0.05). Linear correlation analysis showed that PPARγ protein was positively correlated with γ-GCS activity (r=0.634, P<0.01), and was no significantly correlated with ROS content (r=0.214, P>0.05). PPARγ protein was positively correlated with γ-GCS protein and mRNA (r=0.553, r=0.442, all P<0.01). CONCLUSION: PPARγ activation by RGZ reduces the extent of COPD oxidant/antioxidant imbalance, which plays an important role in the prevention and treatment of COPD. In addition, PPARγ may play an important antioxidant protection role by reducing ROS production, and increasing activity and gene expression of γ-GCS in the lung.
3.The effects of 17-β estradiol on hepatocyte apoptosis and expression of Bcl-2 and Bax in the liver-resected-rats with ischemia reperfusion injury
Fusheng LIN ; Shiqiang SHEN ; Ruicheng YAN ; Ning LI ; Jianchao LIU
Chinese Journal of Hepatobiliary Surgery 2012;18(3):215-219
Objective To investigate the effects of 17-β estradiol on hepatocyte apoptosis and the expression of Bcl-2 and Bax in hepatic tissue after reduced-size ischemia reperfusion injury and its mechanism in liver protection.Methods A rat model of reduced-size hepatic ischemia-reperfusion injury was established in 75 male Sprague-Dawley rats.They were randomly allocated into three groups:Sham group,ischemia-reperfusion(IR)group,and 17-β estradiol(E2 + IR)group.Liver functions,liver histology and hepatocellular apoptosis rates were observed after reperfusion.Hepatocellular ap optosis was determined by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL)method and the expression of Bcl-2 and Bax were determined by Western blotting.Results The levels of ALT and AST were higher and peaked after 12 h of reperfusion in the IR group compared with the sham group.The histological changes in the liver of the IR group consisted of hepatocyte swelling,hepatic sinusoids narrowing,inflammatory cell infiltration and hepatocyte necrosis in some areas of the livers.The IR group also exhibited an increased rate of hepatocellular apoptosis at 12 h after reperfusion.The protein expression of Bcl-2 decreased while the expression of Bax increased.In the 17-β estradiol group,the levels of ALT and AST were lower,the pathological changes were milder and the rate of hepatocellular apoptosis was lower at 12 h in comparison to those of the IR group.The expression of Bcl-2 was higher and the expression of Bax was lower in the 17-β estradiol group in comparison to those of the IR group.Conclusions 17-β estradiol can relieve the hepatic ischemia reperfusion injury in rat livers.17-β estradiol may inhibit apoptosis in hepatic tissue by up regulating Bcl-2 and down-regulating Bax,thus producing a protective effect on hepatic ischemia-reperfusion injury.
4.Effect of liver depression on Th17 and treg in asthma ratsi
Cheng LI ; Aiguo DAI ; Ruicheng HU ; Yongliang JIANG
International Journal of Traditional Chinese Medicine 2012;34(11):989-993
Objective To investigate the effect of liver depression (Liver Qi Stagnation) on Th17,Treg,IL-17,IL-10 and airway inflammation in asthmatic rats,and to clarify the immune mechanism of asthma with liver depression.Methods Established the combined with disease and syndrome model of asthma with liver depression.Collected the bronchoalveolar lavage fluid (BALF) to count the total and differential cell.Lung tissue was observed in microscope; the proportion of Th17 cells and Treg cells of CD4 +T cells in peripheral blood was measured by flow cytometry; the levels of IL-17 and IL-10 were determined by ELISA.Results The total number of inflammatory cells[(96.86±4.43)× 107/L,(88.22±3.22)× 107/L],the proportion of eosinophils [(27.58 ±4.65) %,(22.67±2.43) %],Th17 cells[(6.86±0.98) %,(6.01 ±0.77) %] and IL-17 level [(48.88± 8.06)pg/ml,(43.24± 6.32) pg/ml] of asthma in liver depression group and asthma group were significantly higher than the control group [(30.58 ± 2.49) × 107/L,(0.78 ± 0.12) %,(2.80± 0.82) %,(24.11 ±3.40)pg/ml]; Treg cells [(3.09±0.55) %,(3.96±0.66) %] and IL-10 level [(19.79±2.80) pg/ml,(20.29±3.12) pg/ml] were significantly lower than the control group [(8.02± 1.26) %,(30.79 ± 4.01) pg/ml].The total number of inflammatory cells (96.86 ±4.43) × 107/L,the proportion of eosinophils (27.58±4.65) % and Th17 cells(6.86±0.98) % and IL-17 level (48.88±8.06)pg/mL of asthma in liver depression group were significantly higher than the asthma group (88.22 ± 3.22) × 107/L,(22.67 ± 2.43) %,(6.01 ± 0.77) %,(43.24 ± 6.32) pg/ml;the proportion of Treg cells (3.09 ±0.55)% was significantly lower than the asthma group (3.96± 0.66)%; and the lung histopathology symptoms was more severe than asthma group.Conclusion Liver Qi Stagnation can promote the inflammation of asthma,the imbalance of Th17/Treg and IL-17 level to aggravate the asthma.Liver depression is one of the major internal factors in recurrent episodes of asthma.
5. Application value of detection of 157-114 protein level combined with ADA and CEA levels in pleural effusion in differential diagnosis of pleural effusion
Journal of Jilin University(Medicine Edition) 2020;46(3):614-619
Objective: To detect the levels of B7-H4 protein, adenosine deaminase (ADA) and carcinoembryonic antigen (CEA) in the malignant pleural effusion associated with lung cancer (LC-MPE) and tuberculous pleural effusion (TPK), and to evaluate the values of single and combined detection of B7-H4 protein, ADA and CEA in differential diagnosis of LC-MPE and TPK. Methods: A total of 202 samples of pleural effusion (PE) from 120 LC-MPE patients (LC-MPE group) and 82 TPE patients (TPE group) were collected. The levels of ADA and CEA and B7-H4 protein in PE were detected by the enzymatic, electrochemiluminescence and ELISA methods, respectively; and the receiver operating characteristic curve (ROC curve) was used to determine the diagnostic efficiencies of the above indexes alone or in combination, such as the sensitivity, the specificity, the Yoden index (YI)» and the area under ROC curve ( AUC). Results: The level of ADA in PE of the patients in TPE group was higher than that in LC-MPE group (P<0. 05); the levels of CEA and B7-H4 protein in PE of the patients in LC-MPE group were higher than those in TPE group ( P'-CO. 05). The ROC curve analysis results showed that the sensitivities of single detection of ADA, CEA and B7-H4 protein were 93.30%, 83.33% and 79.90%, respectively; the specificities were 86. 59%, 96.34% and 72. 50%, respectively; the AUC were 0. 927, 0. 925 and 0. 836, respectively. The combined detection of the three indexes had the highest diagnostic value; the sensitivity was 93. 90% , the specificity was 97. 50% , the YI was 0. 971, and the AUC was 0. 998. Conclusion: The combined detection of levels of ADA, CEA and B7-H4 protein in PE is superior to the single evaluation of each index, which can greatly improve the differential diagnosis efficiencies of LC-MPE and TPE.
6.The preparation of recombinant adenovirus Ad-Rad50-GFP and detection of the optimal multiplicity of infection in CNE1 transfected hv Ad-Rad50-GFP.
Ruicheng YAN ; Jiancong HUANG ; Ling ZHU ; Lihong CHANG ; Jingjia LI ; Xifu WU ; Jin YE ; Gehua ZHANG
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2015;29(24):2143-2146
OBJECTIVE:
The optimal multiplicity of infection (MOI) of the recombinant adenovirus Ad-Rad50-GFP carrying a mutant Rad50 gene expression region on the cell growth of nasopharyngeal carcinoma and the viral amplification efficiency of CNE1 cell infected by this adenovirus were studied.
METHOD:
The biological titer of Ad-Rad50-GFP was measured by end point dilution method. The impact of recombinant adenoviral vector transfection on the growth of CNE1 cells was observed by cell growth curve. Transfection efficacy of recombinant adenoviral vector was observed and calculated through fluorescence microscope. The expression f mutant Rad50 in the Ad-Rad50-GFP transfected CNE1 cells with optimal MOI was detected by Western Blot after transfection.
RESULT:
The biological titer of Ad-Rad50-GFP was 1.26 x 10¹¹ pfu/ml. CNE1 cell growth was not influenced significantly as they were transfected by recombinant adenoviral vector with MOI less than 50. Transfection efficacy of recombinant adenoviral vector was most salient at 24 hours after transfection, with the high expression of mutant Rad50, and the efficiency still remained about 70% after 72 hours.
CONCLUSION
Recombinant adenoviral vector Ad-Rad50-GFP could transfect CNE1 cells as well as result in the expression of mutant Rad50 in CNE1 cells effectively. MOI = 50 was the optimal multiplicity of infection of CNE1 cells transfected by recombinant adenoviral vector Ad-Rad50-GFP.
Adenoviridae
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Carcinoma
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Cell Line, Tumor
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Genetic Vectors
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Humans
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Nasopharyngeal Carcinoma
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Nasopharyngeal Neoplasms
;
Transfection
7.Translational medicine used inZhang-Chongquan old TCM inheritance pattern academic experience
Huiping LIU ; Chongquan ZHANG ; Rong YU ; Guomin ZHANG ; Weining ZHANG ; Xianchun PU ; Fenglei WANG ; Zepu NING ; Zhi LI ; Ruicheng ZHAO
International Journal of Traditional Chinese Medicine 2014;(10):932-934
Objective To promote application and explore the old Chinese tradition of academic experience and their experience side mode.Methods In the study, the model of translational medicine was studied combined with the characteristics of Chinese medicine, taken the model from clinical to basic (experimental) to clinical research, combined with prospective and review of the clinical and research methods, with the modern information technology and other technological, and Tonifying Five Internal Organs(TFIO) byZhang-Chongquanfor treatment of coronary heart disease, vascular dementia.Results The article was Clarified the theory of TFIO byZhang-Chongquan,with clinical evidence.Conclusion The model of translational medicine can be used to summarize the academic experience.
8.Detection of carcinoembryonic antigen levels in pleural effusion and serum and their ratio for differential diagnosis of pleural effusion resulting from tuberculosis and lung cancer.
Ruicheng LI ; Zhaowei GAO ; Ke DONG ; Huiping WANG ; Huizhong ZHANG
Journal of Southern Medical University 2019;39(2):175-180
OBJECTIVE:
To study the clinical value of detecting carcinoembryonic antigen levels in pleural effusion (PCEA) and serum (SCEA) and their ratio (P/S) in the differential diagnosis of pleural effusions resulting from tuberculosis and lung cancer.
METHODS:
This retrospectively study was conducted among 82 patients with pleural effusion caused by pulmonary tuberculous (TB; control group) and 120 patients with pleural effusion resulting from lung cancer in our hospital between April, 2016 and March, 2018. PCEA, SCEA and P/S were compared between the two groups and among the subgroups of lung cancer patients with squamous cell carcinoma (SqCa), adenocarcinoma (ACA), small cell carcinoma (SCLC). The receiveroperating characteristic curve (ROC) analysis was used to confirm the optimal critical value to evaluate the diagnostic efficiency of different combinations of PCEA, SCEA and P/S.
RESULTS:
PCEA, SCEA and P/S were significantly higher in the overall cancer patients and in all the 3 subgroups of cancer patients than in the patients with TB ( < 0.05). The areas under the ROC curve of PCEA, SCEA and P/S were 0.925, 0.866 and 0.796, respectively; PCEA had the highest diagnostic value, whose diagnostic sensitivity, specificity, accurate rate, and diagnostic threshold were 83.33%, 96.34, 88.61%, and 3.26 ng/ml, respectively; SCEA had the lowest diagnostic performance; the diagnostic performance of P/S was between that of SCEA and PCEA, but its combination with SCEA greatly improved the diagnostic performance and reduced the rates of misdiagnosis and missed diagnosis. Parallel tests showed that the 3 indexes combined had significantly higher diagnostic sensitivity than each or any two of the single indexes ( < 0.05), but the diagnostic specificity did not differ significantly. The area under the ROC curve of combined detections of the 3 indexes was 0.941 for diagnosis of lung cancer-related pleural effusion, higher than those of any other combinations of the indexes.
CONCLUSIONS
The combined detection of PCEA, SCEA and P/S has a high sensitivity for diagnosis of lung cancer-related pleural effusion and provides important information for rapid and accurate diagnosis of suspected cases.
Carcinoembryonic Antigen
;
analysis
;
blood
;
Case-Control Studies
;
Diagnosis, Differential
;
Humans
;
Lung Neoplasms
;
blood
;
complications
;
Pleural Effusion
;
blood
;
diagnosis
;
immunology
;
Pleural Effusion, Malignant
;
blood
;
chemistry
;
diagnosis
;
ROC Curve
;
Retrospective Studies
;
Sensitivity and Specificity
;
Tuberculosis, Pulmonary
;
complications