1.Enhanced inhibitory effect and mechanisms of adenovirus-mediated hepatocellular carcinoma supressor gene 1 combined with 5-FU on growth of LoVo cells
Jian ZHANG ; Jingying HU ; Xianghua WU ; Weijian GUO
Chinese Journal of Digestion 2010;30(2):90-93
Objective To observe the enhanced inhibitory effect of adenovirus (Ad)-mediated HCCS1 combined with 5-FU on the growth of LoVo cells, and to explore the molecular mechanisms.Methods RT-PCR and Western blot were used to detect the expression of HCCS1 in LoVo cells infected with Ad HCCS1. CCK-8 assay was applied to observe different inhibitory effects of different treatments on growth of LoVo cells. The apoptotic rates were detected by using flow cytometry. The apoptotic proteins were detected by using Western blot. Results ① The recombinant adenovirus, Ad HCCS1, could trigger the expression of HCCS1 in LoVo cell. ② In comparison with controls (92.23%±3.77%), the cell viability rate of LoVo was only (11.23±4.61 )% on 96 h after the combination treatment of 5-FU and Ad-HCCS1 (P<0. 01). ③ The apoptotic rate was (27.57±1.78)% on 72 h after the combination treatment, which was higher than that in 5-FU treated cells (8.64±0.94)%, Ad-HCCS1 treated cells (13.19±1.32)% and 5-FU Ad treated cells (12.16±1.28)%, (P<0. 01). ④ Cathepsin D was only detected in Ad HCCS1-infected cells. When treated with 5-FU, the procaspase-8 was decreased and the cleaved Bid was increased in cytosol. The lowest level of Bax and the highest level of cytoso C and cleaved caspase-3 were detected in cytosols of 5-FU+Ad HCCS1 treated cells. Conclusion The inhibitory and proapoptotic effects are significantly enhanced in LoVo cells when treated with Ad-HCCS1+5-FU. The key protein of the cross-talk is Bax and these data provided a new strategy to treat colorectal carcinomas.
2.T, B and NK lymphocyte subgroups in the pathogenesis of chronic urticaria
Huilan ZHU ; Runxiang LI ; Qing GUO ; Jingying LUO ; Zhenjie LI ; Jiayan LI ; Zhihua WU
Chinese Journal of Dermatology 2008;41(6):391-393
Objective To study the relationship of T, B and NK lymphocytes with the pathogenesis of chronic urticaria. Methods Flow cytometry was applied to assess the proportion of T, B and NK lymphocyte subgroups in the peripheral blood of 51 patients with chronic urticaria and 30 sex and age-matched human controls. The CD4:CD8 ratio was calculated. Moreover, the symptoms, disease course and response to antihistamines of these patients were evaluated by one physician. Results The percentage of CD8+ T and NK cells, CD4:CD8 ratio were (27.20±8.22)%, (21.20±10.84)% and 1.48±0.62, respectively, in these patients,(29.9±3.74)%, (17.5±3.56)%, 1.24±0.27, respectively, in the controls; the differences were significant between the two groups (all P<0.05). Decreased levels of CD3+ T cells, CD8+ T cells and B cells were noted in patients resistant to antihistamines compared with those responsive to antihistamines[(61.81±11.70)% vs (75.74±2.36)%, (24.00±7.79)% vs (34.22±9.30)%, (10.78±2.07)% vs (15.25±4.10)%, P<0.05, 0.01, 0.05, respectively)], while the CD4:CD8 ratio and percentage of NK cells were increased in antihistamine-resistant patients compared to those in antihistamine-sensitive patients [1.67±0.76 vs 1.17±0.41, (28.61±12.62)% vs (12.78±6.02)%, both P<0.01 ]. In these patients with chronic urticaria, the percentages of CD3+ T and CD8+ T cells were negatively correlated with the symptom scores (R = -0.31, -0.28, respectively, both P<0.05 ), while the percentage of B cells was positively correlated with the symptom scores and disease course (R = 0.53, 0.55, respectively, both P<0.01 ). Conclusions There is an abnormality in the proportion of T, B and NK lymphocyte subgroups in patients with chronic urticaria,which indicates that humoral immunity may be involved in the pathogenesis of chronic urticaria and the mechanism for responsiveness to antihistamine.
3.Role of TGF-β1/Smad signaling in angiotensinⅡmediated down-regula-tion of connexin 43
Jingying HOU ; Changqing ZHOU ; Shaoxin ZHENG ; Tianzhu GUO ; Huibao LONG ; Quanhua WU ; Tingting ZHONG ; Tong WANG
Chinese Journal of Pathophysiology 2016;32(10):1729-1736
AIM:To analyze the alterations of angiotensin Ⅱ (Ang Ⅱ), connexin 43 (Cx43), angiotenisinⅡreceptor type 1 (AT1) and signaling molecules in the TGF-β1/Smad pathway in different regions of the left ventricular heart tissue for exploring whether Ang Ⅱregulates Cx43 expression via the TGF-β1/Smad signaling pathway in myocardial infarction ( MI) rats.METHODS:MI was induced in 20 male Sprague-Dawley rats by the left anterior descending coronary artery ligation.The rats were then randomized into 2 groups.In the losartan group, 20 mg· kg-1· d-1 of losartan were ad-ministered for 2 weeks.Heart functions were assessed after surgery and 2 weeks later again following the above treatments . All the rats were sacrificed and relevant molecules , including Ang Ⅱ, AT1, and Cx43 were determined thereafter in diffe-rent areas of the left ventricle .TGF-β1 and its downstream signaling molecules , including Smad 2, Smad 3 and Smad 7, were also detected .RESULTS:In losartan group , both left ventricular internal dimension diastole ( LVIDd) and left ven-tricular internal dimension systole (LVIDs) were smaller, with diminished interventricular septal thickness (IVSd) and left ventricular posterior wall depth ( LVPWd ) and distinct improvement of left ventricular ejection fraction ( LVEF ) ( P<0.05 ) .Losartan therapy exhibited a reduction of Ang Ⅱin the infarct zone and the border zone in the cardiac tissues .AT1 was obviously attenuated in the infarct zone with an enhanced expression of Cx 43, which was also elevated in the border zone and none infarct zone .TGF-β1, Smad 2 and Smad 3 were decreased in different zones of the left ventricle , while Smad 7, in contrary to the above factors , presented a converse alteration .CONCLUSION:The activation of Ang Ⅱpro-vokes downregulation of Cx 43 through TGF-β1/Smad signaling pathway in MI rats .
4.Peroxisome proliferator-activated receptor gamma promotes exogenous mesenchymal stem cells to express connexin 43:its role and mechanism
Ping YAN ; Jingying HOU ; Shaoxin ZHENG ; Huibao LONG ; Changqing ZHOU ; Tianzhu GUO ; Quanhua WU ; Tingting ZHONG ; Tong WANG
Chinese Journal of Tissue Engineering Research 2016;20(23):3357-3365
BACKGROUND:Piglitazone, aperoxisome proliferator-activated receptor γ(PPAR-γ) agonist, has been demonstrated topromote survivalandcardiac differentiation ofexogenous bone marrow mesenchymal stem celsto improvecardiacfunction.In this study, we attempted to investigate whether pioglitazone couldinduce cardiac differentiation of endogenous bone marrow mesenchymal stem celsandimprove cardiacfunction, andmeanwhile, probed into the relevant mechanisms.
OBJECTIVE:To compare the therapeutic efficacy ofpioglitazone combined with bone marrow mesenchymal stem cel transplantation, pioglitazone alone and phosphate buffer solution(PBS)and to investigatetherelevant mechanisms.
METHODS:ThirtySprague-Dawley ratswith myocardial infarctioninducedby ligation of the left anterior descending coronary artery were randomized intocombined group (combination of bone marrow mesenchymal stem cels and pioglitazone), pioglitazone group andPBSgroup. Two weeks later, PKH26-labeled bone marrow mesenchymal stem cels inPBSorPBSalone wereinjected into the local infarct zone in the combinedgroup andthe other twogroups, respectively. Pioglitazone (3 mg/kg/d) was given by the oral gavage in the combinedand pioglitazone groups forcontinuous2weeks after cels transplantation. At 2weeks after treatment, cardiac functions were evaluated. In addition, expressions of PPAR-γ, connexin 43 and relative factors in transforming growth factor-β1/SMAD signaling pathway were examined in different areas of the left ventricle from each harvested heart.
RESULTS AND CONCLUSION:There were no differences in the baseline parameters of cardiac function between the two groups.Twoweeksafter treatment, left ventricular end-diastolic diameter, left ventricular end-systolic diameter and left ventricular ejection fraction were significantlyimprovedin the combined groupcompared with the other two groups; the expression of PPAR-γ was significantly increased in different zones of the left ventriclein the combined andpioglitazone groups.In the combined group, there was a significantlyhigher expression of connexin 43, and the levels of transforming growth factor-β1, SMAD2 and SMAD3 were obviously attenuated in the infarctand marginal zones.However, no differences were found in the abovedeterminants between the pioglitazone andPBSgroups. To conclude, pioglitazone cannot induce the differentiation andproliferation of endogenous bone marrow mesenchymal stem cels, but pioglitazone combined with exogenous bone marrow mesenchymal stem cels can improve cardiac function post myocardial infarction.In this process,PPAR-γmight promote the connexin 43 expression inexogenous bone marrow mesenchymal stem celsviathe blockade oftransforming growth factor-β1/SMAD signaling pathway.
5.Cardiac stem cells improve the electrophysiological stability and ventricular fibrillation threshold via ANGII/AT1R/TGF-beta1/SMAD/CX43 signaling pathway
Ping YAN ; Jingying HOU ; Shaoxin ZHENG ; Huibao LONG ; Tingting ZHONG ; Changqing ZHOU ; Tianzhu GUO ; Quanhua WU ; Tong WANG
Chinese Journal of Tissue Engineering Research 2016;20(28):4226-4233
BACKGROUND:Previous studies have demonstrated that the electrophysiological stability and ventricular fibril ation threshold after myocardial infarction in rats are significantly improved in the mid-term of cardiac stem cel transplantation, but relative regulatory mechanism and pathway remain unclear. OBJECTIVE:To explore the relative molecular regulatory mechanism of cardiac stem cel s improving the electrophysiological stability and ventricular fibril ation threshold after myocardial infarction in rats. METHODS:Myocardial infarction was induced in 20 Sprague-Dawley rats by ligation of the left anterior descending coronary, which were then randomized into two groups (n=10 per group) and were subjected to the injection of cardiac stem cel s labeled with PKH26 in phosphate buffer solution (cardiac stem cel group) or the same amount of phosphate buffer solution (PBS) alone (PBS group) into the local infarct zone at 2 weeks after modeling, respectively. Six weeks later, relevant signaling molecules involved in the ANGII/AT1R/TGF-β1/SMAD/Cx43 pathway were al examined in myocardial tissues of the left ventricle and harvested blood samples. RESULTS AND CONCLUSION:Compared with the PBS group, expressions of connexin 43 in different zones of the left ventricle were significantly increased in the cardiac stem cel group (P<0.01);there was a significant reduction of the angiotensin II level in plasma and different regions of the left ventricular (P<0.05;P<0.01). Furthermore, in the cardiac stem cel group, expressions of angiotensin II type I receptor, transforming growth factor-β1, SMAD2 and SMAD3 were significantly decreased (P<0.01). Whereas SMAD7 was significantly elevated (P<0.05) in different areas of the left ventricle compared with the phosphate buffer solution group. These findings suggest that the cardiac stem cel transplantation can improve the electrophysiological stability and ventricular fibril ation threshold after myocardial infarction by enhancing the expression of connexin 43 via ANGII/AT1R/TGF-beta1/SMAD/CX43 signaling pathway.
6.Clinical significance of MR-guided prostate biopsy based on diffusion-weighted imaging abnormalities for prostate cancer diagnosis in elderly men
Tao GU ; Jingying YU ; Chunmei LI ; Xiaotao DENG ; Zhipeng ZHANG ; Tan GUO ; Jingtao ZHANG ; Hong WANG ; Min CHEN ; Jianye WANG
Chinese Journal of Geriatrics 2017;36(6):673-676
Objective To evaluate the diagnostic efficacy of magnetic resonance(MR)-guided prostate biopsy based on abnormal diffusion weighted imaging for prostate cancer in elder men.Methods From July 2014 to Dec 2016,56 patients (age≥ 65 years) with diffusion weighted imaging (DWI)abnormalities undergoing MR-guided prostate biopsy were retrospectively evaluated.According to pathological diagnosis,patients were divided into prostate cancer and non-prostate cancer groups.The clinical data of two groups were analyzed.The correlation of Gleason score,prostate specific antigen(PSA),and the location and number of biopsy were compared.Results Among 56 cases,32 (57.1%)were diagnosed as prostate cancer,and 24 (42.9%)as chronic prostatitis and prostatic hyperplasia (non-prostate cancer).There were statistically significant differences between prostate cancer and non-prostate cancer in age[(73.0±5.7)vs.(70.1±4.3)year]and PSA[(9.3±6.0)μg/L vs.(6.0± 3.9)μg/L] (both P<0.05),and no statistical differences in biopsy location and biopsy number(t =2.08,2.37,P> 0.05).Gleason score had no correlation with PSA level and biopsy location and biopsy number(r=0.189、-0.183、0.082,P>0.05).Conclusions MR-guided prostate biopsy based on MR-DWI has some merits,such as accurate positioning,fewer numbers of biopsy,and lower false-negative rate.
7.Short-term efficacy of cardiac stem cells in improving the electrophysiological stability and ventricular fibrillation threshold after myocardial infarction
Tingting ZHONG ; Jingying HOU ; Tianzhu GUO ; Shaoxin ZHENG ; Changqing ZHOU ; Huibao LONG ; Quanhua WU ; Hao WU ; Tong WANG
Chinese Journal of Tissue Engineering Research 2015;(28):4537-4543
BACKGROUND:Our previous work demonstrated that cardiac stem cel s (CSCs) transplantation could significantly improve the electrophysiological stability and ventricular fibril ation threshold in rats with myocardial infarction. OBJECTIVE:To compare the influence of CSCs and bone marrow mesenchymal stem cel s (BMSCs) transplantation on the electrophysiological stability and ventricular fibril ation threshold in rats with myocardial infarction in the short term.
METHODS:Thirty male healthy Sprague-Dawley rats were selected to make myocardial infarction models induced by the left anterior descending coronary artery ligation. Then, animals were randomly divided into three groups, CSCs group, BMSCs group and the PBS group, with 10 rats in each group. Two weeks after modeling, animals were respectively given the injection of 5×106 CSCs labeled with PKH26 in 0.1 mL PBS, 5×106 BMSCs labeled with PKH26 in 0.1 mL PBS or 0.1 mL PBS alone into the infracted anterior ventricular free wal . Two weeks after intervention, the electrophysiological characteristics and ventricular fibril ation threshold were measured respectively at the infarct zone, the infarct marginal zone and the non-infarct zone. Labeled CSCs and BMSCs were detected, and the expression of connexin-43 was examined in 5 μm cryostat sections from the infarct marginal zone of each heart.
RESULTS AND CONCLUSION:Compared with the BMSCs group and PBS group, significant differences were revealed in the correct unipolar electrograms activation recovery time dispersion (ARTcd), electrical stimulation-induced malignant ventricular arrhythmias and ventricular fibril ation threshold at the infarct zone, the infarct marginal zone and the non-infarct zone in the CSCs group (P<0.05). Obvious differences were discovered in the ARTcd, electrical stimulation-induced malignant ventricular arrhythmias and ventricular fibril ation threshold on the non-infarct area in the BMSCs group in contrast to the PBS group. Labeled CSCs or BMSCs were identified at the infarct marginal zone and expressed connexin-43. Connexin-43 was abundantly expressed in the CSCs group whereas it was rarely expressed in the BMSCs group, and even not expressed in the PBS group. These findings suggest that CSCs are superior to BMSCs in modulating the electrophysiological stability and the ventricular fibril ation threshold in the short term after transplantation, which is closely correlated with the expression of connexin-43.
8.Identification of targeting relationship between mouse lncRNA-H19 and miR-199 a-5 p by dual-luciferase reporter assay
Jingying HOU ; Changqing ZHOU ; Shaoxin ZHENG ; Tianzhu GUO ; Huibao LONG ; Quanhua WU ; Tingting ZHONG ; Hao WU ; Lei WANG ; Tong WANG
Chinese Journal of Pathophysiology 2016;32(12):2256-2260
AIM:To validate the association between long noncoding (lncRNA)-H19 and microRNA-199a-5p (miR-199a-5p) through the dual-luciferase reporter gene system by construction of a luciferase reporter vector containing the gene of lncRNA-H19.METHODS:The potential complementary binding sites of lncRNA-H19 and miR-199a-5p were predicted by RegRNA 2.0.The H19 gene or its mutant ( Mut) fragment was cloned into luciferase reporter vector psi-CHECK-2.Restriction enzyme analysis and sequence analysis were used to identify whether the recombinant plasmids of the H19 and H19-Mut were successfully constructed .miR-199a-5p mimics, miR-199a-5p inhibitor, miR-199a-5p mimics neg-ative control or miR-199a-5p inhibitor negative control was co-transfected into the 293T cells with the luciferase reporters containing H19 or H19-Mut.Dual-luciferase reporter assay was performed to detect the luciferase activity in different groups in order to verify the relationship between lncRNA-H19 and miR-199a-5p.RESULTS:The results of double enzyme diges-tion and DNA sequencing showed that the sequence of luciferase reporter vector was correct .The results of dual-luciferase reporter assay indicated that the H 19 reporter gene luciferase activity significantly decreased in miR-199a-5p mimics group by 49%(P<0.01), and the H19 reporter gene luciferase activity was obviously upregulated in miR-199a-5p inhibitor group compared with miR-199a-5p mimics group ( P<0.01).However, miR-199a-5p mimics, miR-199a-5p inhibitor, miR-199a-5p mimics negative control and miR-199a-5p inhibitor negative control showed no effect at H 19-Mut reporter gene.CONCLUSION:lncRNA-H19 binds to miR-199a-5p to exert an inhibitory effect at transcriptional level .
9.Pioglitazone administration combined with bone marrow mesenchymal stem cells transplantation improved the heart function of rats with myocardial infarction
Quanhua WU ; Jingying HOU ; Tianzhu GUO ; Tingting ZHONG ; Huibao LONG ; Yue XING ; Changqing ZHOU ; Shaoxin ZHENG ; Tong WANG
Chinese Journal of Tissue Engineering Research 2015;(23):3698-3704
BACKGROUND:Our previous work has demonstrated that bone marrow mesenchymal stem cels (BMSCs) transplantation can improve the heart function of rats with myocardial infarction. However, the overal efficacy is not satisfactory. OBJECTIVE: To adopt pioglitazone as a peroxisome proliferator-activated receptor gamma (PPAR-γ) agonist combined with BMSCs transplantation therapy, thereby further improving cardiac function of rats with myocardial infarction as wel as investigating the relevant mechanisms. METHODS:Twenty Sprague-Dawley rats with myocardial infarction were induced by the left anterior descending coronary artery ligation. The animals were randomized into two groups: BMSCs and BMSCs+pioglitazone. Two weeks later, al the animals received the injection of BMSCs labeled with PKH26 in PBS into the local infarct zone, and then pioglitazone (3 mg/kg/d) was given by the oral gavage for 2 weeks in the BMSCs+pioglitazone group after the cel transplantation. After 2 weeks of cel transplantation, cardiac functions were evaluated by echocardiography. The expressions of PPAR-γ, Connexin 43 and molecules in TGF-β1/SMAD signaling pathway were examined in different areas of the left ventricle from each harvested heart using immunofluorescent staining, western blot assay and qRT-PCR. RESULTS AND CONCLUSION:There were no differences in the baseline parameters of cardiac function between the two groups. At 2 weeks after cel transplantation, the left ventricular internal diameter at end-diastole, left ventricular internal diameter at end-systole and left ventricular ejection fraction were significantly improved in the BMSCs+ pioglitazone group; the expressions of PPAR-γ and Connexin 43 were distinctly increased in different zones of the left ventricle; the levels of TGF-β1, SMAD2 and SMAD3 were obviously attenuated in the infarct zone and border zone. The above-mentioned findings suggest that pioglitazone, a PPAR-γ agonist, can enhance BMSCs potential in improvingthe heart function after myocardial infarction, and PPAR-γ may elevate the expression of Connexin 43via the blockade of the TGF-β1/SMAD signaling pathway in the procedure.
10.Research progress of hypoxia-inducible factor-prolyl hydroxylase inhibitors in the treatment of renal anemia and ischemic heart disease
Haoqi SUN ; Zhentao GUO ; Xiangming WANG ; Jingying WANG
Clinical Medicine of China 2022;38(5):475-480
The hypoxia-inducible factor is a significant regulator of adaptive transcriptional response in hypoxia or hypoxia. Hypoxia-inducible factor (hypoXIA-inducible factor) loses its activity after hydroxylation by proline hydroxylase in a normoxic environment. Proline hydroxylase inhibitor is a kind of new small molecule oral preparation by inhibiting the proline hydroxylase, reducing the degradation of HIF, activating the hypoxia-induced way, adjusting including stimulates erythropoiesis, iron absorption and mobilization, angiogenesis, lipid, and glucose metabolism, inflammation, energy metabolism, cell growth and differentiation, and other physiological reaction, Showed more clinical benefits. In recent years, the application of proline hydroxylase inhibitors in the field of renal anemia has achieved apparent efficacy, and the research in the field of ischemic heart disease has also made significant progress in the future in the treatment of ischemic heart disease and other aspects of good application prospects.