1.Effect of microbubbles mediated ultrasound insonation on proliferation and apoptosis of vascular smooth muscle cells in different phases of ceil cycle
Ping ZHANG ; Yunhua GAO ; Ping LIU ; Zheng LIU ; Kaibin TAN
Chinese Journal of Ultrasonography 2009;18(6):533-536
Objective To investigate the effect of microbubbles mediated ultrasound insonation on proliferation and apoptosis of vascular smooth muscle cells (VSMCs) in different phase of cell cycle. Methods Rat thoracic aortic VSMCs were cultured in vitro by the method of tissue adherence. The cells were synchronized by the methods of serum starvation and double thymidine block. The synchronization results were detected by flow eytometer. VSMCs in different phases of cell cycle were exposed to 1 MHz continuous waves ultrasound for 120 s at intensity 0.3 W/cm2 in the presence of lipid-coated microbubbles (1 ml/L). Apoptosis of VSMCs was analyzed by AnnexinV/PI staining using flow eytometry. The proliferation and the proliferating cell nuclear antigen(PCNA) protein expression of VSMCs were detected by MTT assay and immunoeytochemistry, respectively. Results The synchronized G0/G1 and S phase VSMCs were achieved, with synchronized rates to 89.53 % and 66.87 %, respectively. Ultrasound sonication for 120 s with microbubbles could significantly inhibit the proliferation and downregulate the PCNA expression of S phase VSMCs,but the proliferation and PCNA expression of G0/G1 phase VSMCs were not affected. After treatment of ultrasound with microbubbles, the apoptotic ratio were found to reach (7.05 ± 2.04)% in G0/G1 phase VSMCs and (27.01 ±3.87)% in S phase VSMCs. Conclusions Microbubbles mediated ultrasound insonation can significantly inhabit the proliferation and induce apoptosis in VSMCs at proliferation stage.
2.Effect of ultrasound irradiation combined with liposome membrane microbubbles on the reorgnization of cytoskeleton in vascular smooth muscle cells
Ping ZHANG ; Yunhua GAO ; Zheng LIU ; Ping LIU ; Kaibin TAN
Chinese Journal of Ultrasonography 2009;18(5):444-447
Objective To investigate the effect of ultrasound irradiation combined with liposome membrane microbubbles on the reorgnization of cytoskeleton in vascular smooth muscle cells (VSMCs). Methods Rat thoracic aortic VSMCs were cultured in vitro. VSMCs were exposed to 1 MHz continuous waves ultrasound radiation for 120 s at intensity 0.3 W/cm2in the presence of liposome membrane microbubbles (1 μl/ml) after treated with platelet derived growth factor-BB (PDGF-BB). The reorganizations of microfilaments, microtubules and intermediate filaments were examined by using immunofluorescence and fluorocytochemistry techniques. Results There was a substantial increase in the expression of F-actin and assembly of long bundles of stress fibers in the transversed cell body when treated with PDGF-BB. Neither alterations of β-tubulin nor of vimentin cytoskeletal protein organization were observed in PDGF-BB treated cells as compared to those of the contol group. After ultrasound irradiation combined with liposome membrane microbubbles, the expression of F-actin, β-tubulin and vimentin were reduced along with the simultaneous changes in microfilaments, microtubles and intermediate filaments array. Conclusions Ultrasound irradiation combined with liposome membrane microbubbles can induce significant changes in cytoskeleton structure of VSMCs cultured in vitro.
3.The efficacy of microbubbles augmentation thrombolysis using low-frequency ultrasound: an in vitro study
Yali XU ; Zheng LIU ; Yunhua GAO ; Kaibin TAN
Chinese Journal of Ultrasonography 1993;0(02):-
Objective To promote in vitro tissue-plasminogen activator(t-PA)thrombolysis by using microbubbles augmented low frequency ultrasound(US) insonication.Two kinds of microbubbles,albumin-coated vs.lipid-coated were compared.Methods Human blood clot weighted 200~300 mg was made from of 0.8 ml fresh blood.Twenty kHz ultrasound,lipid and albumin-coated microbubbles were applied to separate clots groups with or without t-PA administration.The clots before and after processing were weighed and then the clot dissolution ratios were calculated.Results The simple ultrasound group and the basic t-PA group dissolved((24.72)?(4.83))% and((35.66)?(3.34))% of the clots,respectively.The clot dissolution ratio rised to((42.06)?(4.20))% when coordinated US and t-PA(P(0.05)).Conclusions The in vitro US augmentation of t-PA thrombolysis can be significantly promoted by introduction of microbubbles.But there is no significant thrombolysis difference between albumin and lipid-coated microbubbles.
4.Optimization of diagnosis indicator selection and inspection plan by 3.0T MRI in breast cancer
Zhongbiao JIANG ; Yunhua WANG ; Zhong HE ; Lejun ZHANG ; Kai ZHENG
Journal of Central South University(Medical Sciences) 2013;38(8):830-837
Objective:To optimize 3.0T MRI diagnosis indicator in breast cancer and to select the best MRI scan program.
Methods:Totally 45 patients with breast cancers were collected, and another 35 patients with benign breast tumor served as the control group. All patients underwent 3.0T MRI, including T1-weighted imaging (T1WI), fat suppression of the T2-weighted imaging (T2WI), diffusion weighted imaging (DWI), 1H magnetic resonance spectroscopy (1H-MRS) and dynamic contrast enhanced (DCE) sequence. With operation pathology results as the gold standard in the diagnosis of breast diseases, the pathological results of benign and malignant served as dependent variables, and the diagnostic indicators of MRI were taken as independent variables. We put all the indicators of MRI examination under Logistic regression analysis, established the Logistic model, and optimized the diagnosis indicators of MRI examination to further improve MRI scan of breast cancer.
Results:By Logistic regression analysis, some indicators were selected in the equation, including the edge feature of the tumor, the time-signal intensity curve (TIC) type and the apparent diffusion coeffcient (ADC) value when b=500 s/mm2. hTe regression equation was Logit (P)=-21.936+20.478X6+3.267X7+21.488X3.
Conclusion:Valuable indicators in the diagnosis of breast cancer are the edge feature of the tumor, the TIC type and the ADC value when b=500 s/mm2. Combining conventional MRI scan, DWI and dynamic enhanced MRI is a better examination program, while MRS is the complementary program when diagnosis is diffcult.
5.Correlation of peripheral blood CD34+ cell level with cerebrovascular disease risk factors in patients with cerebral infarction
Le FANG ; Jiajun CHEN ; Xuxin YANG ; Yiyan YANG ; Xuebing ZHENG ; Shuang ZHANG ; Yunhua ZHAO
Chinese Journal of Geriatrics 2011;30(8):636-639
Objective To observe the change of peripheral blood CD34+ cell level in patients with acute cerebral infarction, and explore its relationships with cerebrovascular risk factors,neurological function and carotid artery intima-media thickness (IMT). Methods The 45 patients with acute cerebral infarction (onset within 72 h) (infarction group) and 27 patients with cerebr ovascular risk factors but without cerebral infarction (high-risk group) were chosen for the study. The cerebrovascular disease risk factors including history of alcohol abuse, smoking, coronary heart disease, hypertension, diabetes, abnormal levels of serum triglycerides, total cholesterol,low-density lipoprotein cholesterol (LDL-C), and high density lipoprotein cholesterol (HDL-C) were recorded in all subjects. The peripheral blood CD34+ cell levels were measured by flow cytometry.The correlations of peripheral blood CD34+ cell level with cerebrovascular disease risk factors were analyzed. The neurological function and carotid artery IMT were recorded in infarction group, and the correlations of peripheral blood CD34+ cell level with neurological function and carotid artery IMT were analyzed. Results (1) The peripheral blood CD34+ cell level was significantly negatively correlated with coronary heart disease, hypertension, diabetes and LDL-C level (r =- 0. 749,-0. 717, - 0. 688, - 0. 764, all P<0. 01) ; (2) Multiple linear regression analysis showed that peripheral blood CD34+ cell level was an independent relative factor of acute cerebral infarction (P<0.05); (3) The peripheral blood CD34+ cell level was lower in infarction group than in high-risk group, and was significantly negatively correlated with neurological deficit score (r=-0. 721, P<0.01) and carotid artery IMT (r= -0. 695, P<0. 01). Conclusions Peripheral blood CD34+ cell level could be an independent relative factor of acute cerebral infarction; The peripheral blood CD34+ cell level is significantly negatively correlated with neurological function and carotid artery IMT in patients with acute cerebral infarction; And it can be used as cytological marker which reflect early vascular endothelial function in patients with ischemic stroke.
6.Mechanism of impact of transcranial ultrasound contrast imaging on the blood-brain barrier: an earlier research
Ping LIU ; Kaibin TAN ; Lihui MA ; Zheng LIU ; Ping ZHANG ; Yunhua GAO
Chinese Journal of Ultrasonography 2009;18(2):159-162
Objective To investigate the mechanism of the increased permeability of the blood-brain barrier during transcranial ultrasound contrast imaging. Methods Sprague-Dawhy (SD) rats were performed transcranial ultrasound contrast imaging, the lanthanum nitrate and the evans-blue were used as tracers,the distribution of the tracers were observed and the transports mechanism were also investigated. Results The opening of the tight junction and increased permeability of the cellular membrane were observed after the transcranial ultrasound contrast imaging. Conclusions The main mechanism of the increased permeability of the blood-brain barrier was the opening of the tight junction and increased permeability of the cellular membrane.
7.Diagnosis and treatment of 47 cases of primary splenic tumors
Jianbao ZHENG ; Xuejun SUN ; Mao MA ; Yunhua WU ; Qi GAO ; Jingsen SHI
Chinese Journal of Hepatobiliary Surgery 2015;21(12):833-835
Objective To investigate the clinical characteristics, diagnosis and treatment of primary splenic tumors.Methods The clinical data of 47 patients with spleen tumors treated in the First Affiliated Hospital of Medical College, Xi'an Jiaotong University from January 2008 to October 2014 were retrospectively analyzed.Results 28 patients had no symptoms and only on physical examination was a mass found in some patients.There were 12 patients who manifested with upper abdominal discomfort or pain, 2 patients with an epigastric mass, l patient manifested with fever and 2 patients manifested with nausea and vomiting.Preoperative examination showed anemia in 7 patients.Three patients manifested with hypersplenism.Preoperative ultrasonic examination was performed on 39 patients with a diagnostic rate of 89.7%.43 patients underwent CT examination which had a diagnostic rate of 90.7%, MRI was performed in 4 patients and the diagnoses were all correct.Of the 47 patients with splenic tumor, 38 patients had a benign tumor.34 patients were treated by surgery, including 22 patients who underwent open splenectomy, 9 patients laparoscopic splenectomy, 3 patients laparoscopic fenestration of splenic cyst.Malignant tumors were found in 9 patients.Four patients underwent splenectomy, 2 patients were treated by laparoscopic splenectomy.Preoperative examination and postoperative pathological examination showed a benign tumor in 38 patients, including 1 1 patients with a splenic cyst, 6 patients with a cavernous hemangioma, 5 patients with an inflammatory pseudo tumor of spleen (accounting for 57.3% of all the benign tumor).Malignant tumors included 5 patients with malignant lymphoma, 1 patient with splenic angiosarcoma, 1 patient with gastric carcinoma which metastasizes to the spleen, 1 patient with cervical carcinoma metastasizing to the spleen and 1 patient with liver carcinoma metastasizing to the spleen.Adjuvant chemotherapy and (or) radiotherapy after surgery were performed for patients with malignant lymphoma of the spleen.For 2 patients who were diagnosed early, surgery combined with radiotherapy or chemotherapy and immunotherapy were alive for more than 18 months.Conclusions The clinical manifestations of splenic tumors lack specificity.The diagnosis mainly depends on ultrasonic examination, CT and MRI.Early diagnosis, radical operation and comprehensive treatment are important to improve the prognosis of patients with malignant tumors of the spleen.
8.Changes and clinical significance of peripheral blood CD34+ cells in patients with acute cerebral infarction
Jiajun CHEN ; Xuexin YANG ; Xuebing ZHENG ; Lihong HUANG ; Shuang ZHANG ; Yunhua ZHAO ; Xiya JIN ; Yuanbing LI
Chinese Journal of Geriatrics 2012;31(11):991-993
Objective To determine the level of peripheral blood CD34 positive (CD34+) cells in patients with acute cerebral infarction (ACI),and to explore its clinical significance.Methods The level of peripheral blood CD34+ cells was determined by flow cytometry within 72 hours of onset of patients with acute cerebral infarction (infarct group,n=45),cerebrovascular risk factors in patients without cerebral infarction (high risk group,n=27) and healthy subjects (control group,n=20).The neural function defect score,infarction lesion volume and carotid artery intima-media thickness (IMT) were determined in patients with infarction group.Results The percentages of peripheral blood CD34 cells in infarction group (0.034 ±0.012)% and the high risk group of patients (0.047±0.009)% were lower than that of control group(0.063±0.009)%,and were lower in infarction group than in high-risk groups (all P<0.05).The percentages of peripheral blood CD34+cells were significantly decreased compared with control group (P<0.05) in infarction patients with mild [(0.047±0.009)%],moderate [(0.036±0.009)%],severe [(0.022±0.007)%] infarction nervous function defect score.Wherein,the percentages were lower in severe group than in the moderate group,moderate group was lower than in mild group (all P<0.05).The percentages of peripheral blood CD34 cells in infarction patients with small,moderate,large infaret lesion volume were lower than in control group (P<0.05),wherein,were lower in large group than in moderate group,lower in moderate group than in treatment group (all P<0.05).Infarction patients were confirmed with carotid atherosclerosis (CAS) by carotid ultrasound.The extent of lesion were divided into carotid artery intimal thickening group [(0.043±0.010)%],carotid artery plaque group [(0.036±0.010)%],and carotid artery stenosis group [(0.023±0.009)%].The levels of peripheral blood CD34+ cells in three groups of patients were decreased compared with control group.The levels were lower in carotid artery stenosis group than in carotid artery plaque group,lower in carotid artery plaque group than in carotid artery intimal thickening group (all P<0.05).Conclusions The level of peripheral blood CD34+ cells in acute cerebral ischemia is reduced,it can become a sensitive and early indicator of cerebral ischemia,and its level is related to neurologic impairment,infarction size and the degree of carotid artery atherosclerosis.
9.The anti-HIV activity of three 2-alkylsulfanyl-6-benzyl-3, 4-dihydropyrimidin-4 (3H)-one derivatives acting as non-nucleoside reverse transcriptase inhibitor in vitro.
Jing LONG ; Dehua ZHANG ; Gaohong ZHANG ; Zhikun RAO ; Yunhua WANG ; Siucheung TAM ; Yanping HE ; Yongtang ZHENG
Acta Pharmaceutica Sinica 2010;45(2):228-34
It was recently shown that several new synthetic 2-alkylsulfanyl-6-benzyl-3, 4-dihydropyrimidin-4(3H)-one (S-DABO) derivatives demonstrated anti-HIV-1 activity. Three of the derivatives namely RZK-4, RZK-5 and RZK-6 were used in this study to explore their inhibitory effects on a variety of HIV strains. These compounds at a concentration of 200 microg mL(-1) almost completely inhibited the activity of recombinant HIV-1 reverse transcriptase. All of the three compounds reduced replication of HIV-1 laboratory-derived strains, low-passage clinical isolated strain, and the drug resistant strain. In particular RZK-6 showed potent activity against the HIV-1 drug resistant strain. In general, the antiviral activities are similar in magnitude to nevirapine (NVP), which is a non-nucleoside reverse transcriptase inhibitor approved by FDA. The therapeutic indexes of these compounds were remarkable, ranging from 3704 to 38462 indicating extremely low cytotoxicity. These results suggest that the three S-DABO derivatives in this study have good potential for further development in anti-HIV-1 therapy. It may be particularly useful to target at the non-nucleoside reverse transcriptase inhibitors resistant HIV-1 strain.
10.Mesenchymal stem cells transplantation under the diagnostic ultrasound mediated microbubbles destruction in improving myocardial perfusion after rabbit myocardial ischemia
Yali XU ; Yunhua GAO ; Zheng LIU ; Kaibin TAN ; Zhenqiang FANG ; Yongliang LIU ; Pin QIAN ; Xi YANG
Chinese Journal of Ultrasonography 2009;18(9):792-796
Objective To explore the value of diagnostic ultrasound mediated microbubble destruction in improving the myocardial perfusion and left ventricular systolic function when cooperated with the mecsenchymal stem cells(MSCs) transplantation in rabbit myocardial ischemia. Methods One week after myocardial ischemia (MI) modeling,36 rabbits were divided into 3 groups,the control group(group Ⅰ) ,intravenous injection of MSCs group(group Ⅱ) and ultrasound + microbubble + MSCs group (group Ⅲ). Myocardial contrast enhancement (MCE) was performed and quantification analysis of anterior wall was assessed with Photoshop. Left ventrieular systolic function was assessed with M-mode echocardiography and bi-plane Simpson's method. CD34 expression in heart was detected with immunohistochemisty(IHC). Western blotting was applied to detect the level of VEGF in three groups. Results The differences of gray scale analyzed with histogram of Photoshop in anterior wall of ischemia myocardium between the group Ⅰ and group Ⅱ or group Ⅲ were significant,and P value was 0. 032 and 0. 000 , respectively. There were significant differences of FS between group Ⅲ (30. 43±4.09)% and group Ⅱ (26.29±2.93)%, P<0.01, and similar to group Ⅰ (19.28 ± 2.84)%. The difference of EF(%) between group Ⅲ and group Ⅱ was significant [(61.5±5.8 vs 53.6±4. 71), P<0. 05] ,or markedly significant between group Ⅲ and group Ⅰ [(61.5±5.8 vs 42.6± 5.0), P <0.01]. EF(%) assessed with bi-plane Simpson's method was significantly increased from (34.64 ± 4.59) in group Ⅰ to (41.78 ± 4.21) in group Ⅱ and (48.6±3.96) in group Ⅲ. The expression of CD34 assessed with immunohistochemistry was the highest in group Ⅲ. The level of VEGF with western blotting in group Ⅲ was significantly higher than other two groups. Conclusions It is an efficacious transplantation means of MSCs infusion under the ultrasound mediated microbubles destruction in improving the myocardial perfusion and cardiac systolic function.