1.Optimization of low-power ultrasound with micro-bubbles in inducing early apoptosis of DU145 cells
Yanduan, LIN ; E, SHEN ; Shuliang, NAN ; Bing, HU
Chinese Journal of Medical Ultrasound (Electronic Edition) 2014;(3):67-70
Objective To optimize the parameters of the low-frequency/low-energy ultrasound combined with micro-bubbles in inducing early apoptosis of DU145 cells (an androgen-independent prostatic cancer cells). Methods In our study, the impact of ultrasonic power, micro-bubbles/cell suspension volume rate and irradiation time were investigated. Three levels of each factor were deifned as ultrasonic power (60, 80, 100 mW), micro-bubbles/cell suspension volume rate (10%, 20%, 30%), irradiation time (30, 60, 90 s). According to the three-factor three-level orthogonal design, nine experiments were carried out. The early apoptosis was detected by lfow cytometry. A new experiment was designed with the optimized parameters. Another group without ultrasound irradiation was designed as the control group. Flow cytometry and transmission electron microscope (TEM) were used to detect the early apoptosis. Results In descending order, the inlfuence of these factors on the cell early apoptosis were:ultrasonic power>micro-bubbles/cell suspension volume rate>irradiation time. Moreover, the inlfuence of each factor level were:80 mW>60 mW>100 mW in ultrasonic power, 20%>30%>10%in micro-bubbles/cell suspension volume rate, 60 s>90 s >30 s in irradiation time. The early apoptosis rate of experiment group was 10.41%, while the control group was 0.94%. TEM showed apoptotic cells in the experiment group. Conclusions The optimized parameter of low-frequency/low-energy ultrasound with micro-bubbles in inducing early apoptosis of DU145 cells are ultrasonic power of 80 mW, micro-bubbles/cell suspension volume rate of 20%, and irradiation time of 60 s. With the optimized parameters, the early apoptosis rate of the experiment group has signiifcant higher than the control group.
2.Experimental study of low-frequency ultrasound combined with microbubbles promote enhanced green lfuorescent protein plasmid transfecting into transplanted prostate cancer of nude mice
Shuliang, NAN ; E, SHEN ; Yanduan, LIN ; Wenkun, BAI ; Bing, HU
Chinese Journal of Medical Ultrasound (Electronic Edition) 2014;(5):434-438
Objective To investigate the feasibility of low-frequency ultrasound combined with microbubbles improving transfection of enhanced green lfuorescent protein plasmid (pEGFP) to subcutaneously transplanted prostate cancer in nude mice, and to optimize the parameters of intensity. Methods The model of nude mice of subcutaneously transplanted prostate cancer were built. Then they were divided into 2 groups including low-frequency ultrasound group and low-frequency ultrasound combined with microbubbles group, each group with 15 mice. Then 250μl of mixture of saline and pEGFP (50μg) were co-injected in low-frequency ultrasound group and 250μl of mixture of SonVue and pEGFP (50μg) were co-injected in low-frequency ultrasound combined with microbubbles group through tail vein. According to intensity, they were divided into 3 subgroups respectively, including group 1 W/cm2, group 2 W/cm2 and group 3 W/cm2, each group with 5 mice. Ultrasound was applied at 80 kHz input frequency with 50%duty cycle for 10 minutes after pEGFP injection. The tumors were collected on the third day after treatment. The expression of pEGFP in tumor was examined by laser scanning confocal microscope (ISCM) and the average lfuorescence intensity were estimated. At the same time, routine pathological examination was performed. The mean lfuorescence intensity of low frequency ultrasound group and low frequency ultrasound combined with microbubble group were compared with one-way ANOVA and those of any two groups were compared by SNQ-q test. Results TThe mean lfuorescent light in low-frequency ultrasound combined with microbubbles group were 23.75±2.54, 30.25±1.67 and 59.60±2.03, which were obviously stronger than those of low-frequency ultrasound treatment group (14.04±1.35, 14.66±0.98 and 14.32±1.20), the difference was statistically signiifcant (the value of q were 18.26, 14.12 and 13.88, P<0.05). The rate of gene transfection increased along with the enhancement of the ultrasound intensity (the value of q were 15.33, 17.81 and 13.79, P<0.05). Histology analyses performed by HE staining showed that there was no damage to the tumor tissues in all groups, tumor tissues were intact and without infection. Conclusions Low-frequency ultrasound combination with microbubbles can significantly enhance pEGFP transfecting into subcutaneously transplanted prostate cancer in nude mice. In certain range, improving the ultrasound intensity can increase the rate of gene transfection.
3.Relationship between myocardial glucose metabolism in right heart and cardiac function in patients with idiopathic dilated cardiomyopathy
Xinghong MA ; Lei WANG ; Liwei XIANG ; Yong YANG ; Shuliang HU ; Shihua ZHAO ; Wei FANG
Chinese Journal of Nuclear Medicine and Molecular Imaging 2015;35(2):81-84
Objective To explore the relationship between myocardial glucose metabolism in the right heart and cardiac function in patients with idiopathic dilated cardiomyopathy (IDCM).Methods From October 2010 to May 2013,64 consecutive patients (45 males,19 females; mean age:(53 ± 12) years) with IDCM were enrolled in this study.All patients underwent 18F-FDG myocardial PET imaging,late-gadolinium enhancement cardiac MRI (cMRI-LGE) and UCG.TrueD software was used to determine the SUVmax in right (SUV-R) and left (SUV-L) ventricular free wall,and the ratio of SUV-R to SUV-L (SUV-R/L) was calculated.The cMRI-LGE was used to evaluate LVEF,RVEF and LGE in left heart.Pulmonary artery systolic pressure (PASP) was measured by UCG.Two-sample t test and Pearson correlation analysis were used to analyze the data.Results Both SUV-R and SUV-R/L had negative correlation with LVEF (r=-0.48,-0.33; both P<0.01) and also with RVEF (r=-0.45,-0.44; both P<0.01).SUV-R/L positively correlated with PASP (r =0.58,P<0.01).SUV-R/L of patients with and without LGE were statistically different (0.45±0.18 vs 0.36±0.12; t=2.50,P<0.05).SUV-R and SUV-L of patients with LGE were 2.0±0.9 and 4.7±1.6,and those of patients without LGE were 1.7±0.9 and 5.0±2.8.Both differences of SUV-R and SUV-L between the two groups were not statistically significant (t=0.54 and 0.63,both P>0.05).Conclusions Myocardial glucose metabolism in right heart is closely correlated with left and right ventricular function.The assessment of 18F-FDG uptake in right heart by PET imaging is potentially useful for evaluation of the severity and prognosis in IDCM patients.
4.Elevated level of renal xanthine oxidase mRNA transcription after nephropathogenic infectious bronchitis virus infection in growing layers.
Huayuan LIN ; Qiqi HUANG ; Xiaoquan GUO ; Ping LIU ; Weilian LIU ; Yuelong ZOU ; Shuliang ZHU ; Guangfu DENG ; Jun KUANG ; Caiying ZHANG ; Huabin CAO ; Guoliang HU
Journal of Veterinary Science 2015;16(4):423-429
To assess relationships between xanthine oxidase (XOD) and nephropathogenic infectious bronchitis virus (NIBV) infection, 240 growing layers (35 days old) were randomly divided into two groups (infected and control) of 120 chickens each. Each chicken in the control and infected group was intranasally inoculated with 0.2 mL sterile physiological saline and virus, respectively, after which serum antioxidant parameters and renal XOD mRNA expression in growing layers were evaluated at 8, 15 and 22 days post-inoculation (dpi). The results showed that serum glutathione peroxidase and superoxide dismutase activities in the infected group were significantly lower than in the control group at 8 and 15 dpi (p < 0.01), while serum malondialdehyde concentrations were significantly higher (p < 0.01). The serum uric acid was significantly higher than that of the control group at 15 dpi (p < 0.01). In addition, the kidney mRNA transcript level and serum activity of XOD in the infected group was significantly higher than that of the control group at 8, 15 and 22 dpi (p < 0.05). The results indicated that NIBV infection could cause the increases of renal XOD gene transcription and serum XOD activity, leading to hyperuricemia and reduction of antioxidants in the body.
Antioxidants
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Chickens
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Glutathione Peroxidase
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Hyperuricemia
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Infectious bronchitis virus*
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Kidney
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Malondialdehyde
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RNA, Messenger*
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Superoxide Dismutase
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Uric Acid
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Xanthine Oxidase*
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Xanthine*
5.Early and mid-term results of surgical treatment for complete atrioventricular septal defect
Fengxiang LI ; Minghui ZOU ; Yanqin CUI ; Li MA ; Xu ZHANG ; Shuliang XIA ; Chunmei HU ; Xinxin CHEN
Chinese Journal of Thoracic and Cardiovascular Surgery 2023;39(7):398-404
Objective:To summarize the results of surgical treatment for complete atrioventricular septal defect(CAVSD) in early and middle stages.Methods:147 children with CAVSD in Guangzhou Women and Children’s Medical Center from January 2010 to December 2019 were selected, Males 85, females 62, median age of surgery 5 months(1 months-10 years old), median body mass 5.5 kg(2.4-20.9 kg). Complete atrioventricular septal defect was diagnosed by ultrasonic cardiogram before surgery. All the children underwent atrial ventricular valve formation and underwent simultaneous repair.Outpatient follow-up was planned.Ultrasonic cardiogram and electrocardiogram were performed. SPSS 22 statistical software was used for data analysis.Results:All 147 CAVSD patients underwent one-time surgical correction.Early postoperative death occurred in 7 cases(4.76%). The causes of death were: 3 cases of pulmonary hypertension crisis, 3 cases of severe mitral insufficiency(MI), 1 case of postoperative malignant arrhythmia, and the rest of the children were cured and discharged. Permanent pacemaker was installed in 3 patients due to atrioventricular block(AVB). The follow-up time was 1-10 years old, and 2 patients died in late stage: 1 patient did not seek medical treatment in time due to infection, and 1 patient had unknown cause. Five patients underwent secondary surgery: 4 due to severe mitral/tricuspid insufficiency(MI/TI) and 1 due to delayed AVB. The mid-term follow-up showed 9 cases of severe MI and 4 cases of severe TI. Compared with children with surgical age<3 months and ≥3 months, there were statistically significant differences in postoperative ventilator-assisted ventilation time, severe MI before postoperative discharge and total mortality between the two groups( P<0.05). Mid-term follow-up results showed no difference between the two groups. There were statistically significant differences in surgical age, postoperative CICU stay time and total hospital stay between the children with trisomy 21-syndrome and those without trisomy 21-syndrome( P<0.05), and there was no difference between the two groups in mid-term follow-up results. Residual shunt of 1-3 mm VSD was found in 29 cases, 26 cases were closed during follow-up, and 3 cases had smaller residual shunt. Conclusion:Modified single patch technique treatment of CAVSD has good effect, low mortality and low re-operation rate. But age <3 months group, infant mortality was significantly increased, the duration of postoperative mechanical assisted ventilation was prolonged, and the proportion of early postoperative severe MI was high.Severe MI and TI is easy to occur after CAVSD, which requires long-term follow-up and timely treatment. The children with trisomy 21-syndrome were similar to those with normal chromosome except for longer stay in ICU and total hospital stay.
6.MicroRNA-182 modulates high glucose-induced cardiomyocyte hypertrophy via targeting Rac1.
Zheying MENG ; Yu WANG ; Yanduan LIN ; Shuliang NAN ; Weiping XU ; Bing HU ; E SHEN ; Email: SJLYXZCS@163.COM.
Chinese Journal of Cardiology 2015;43(7):619-624
OBJECTIVETo investigate the role and signalling of microRNA(miR)-182 on regulating high glucose-induced cardiomyocyte hypertrophy.
METHODSThe candidates of miR which might potentially be involved on targeting Rac1 were predicted by applying bioinformatics analysis. The expression of all related candidates miRs was verified by real-time reverse transcription-PCR (RT-PCR) in cardiac tissues of db/db mice and db/m mice. Then the relationship between candidates miR and Rac1 was investigated with Pearson relevant analysis. Neonatal mice cardiomyocytes were cultured and divided into 2 groups: normal glucose group and high glucose group. The level of selected miR and Rac1 in two groups was detected by RT-PCR. Neonatal mice cardiomyocytes were then randomly divided into 4 groups: normal glucose group, selected microRNA mimics control group, high glucose group, high glucose plus selected miR mimics control group. The morphology of cardiomyocyte in each group was detected under light microscope. Furthermore, Rac1, β-MHC and α-SMA expressions were detected in cultured cardiomyocyte treated by high glucose for 48 h after transfecting selected miR mimics by RT-PCR and Western blot.
RESULTSA total of 6 miR candidates potentially targeting Rac1 were screened by bioinformatics, which were miR-182, miR-142-3p, miR-140, miR-101a, miR-429 and miR-200b. Among these candidates, miR-182 and miR-142-3p expression was significantly downregulated in cardiac tissues of db/db mice compared with db/m controls (P < 0.05). MiR-182 was negatively correlated with Rac1 by person analysis (r = -0.891 02). Downregulation of miR-182 and upregulation of Rac1, β-MHC, α-SMA were found in high glucose-induced cardiomyocyte. After transfection of miR-182 mimics, hypertrophic changes were significantly reduced and Rac1 as well β-MHC expression was significantly downregulated in cardiomyocyte incubated with high glucose.
CONCLUSIONMiR-182 might be involved in the regulation of high glucose-induced myocardial hypertrophy process via targeting Rac1.
Animals ; Cardiomyopathy, Hypertrophic ; metabolism ; Down-Regulation ; Glucose ; physiology ; Mice ; MicroRNAs ; physiology ; Myocytes, Cardiac ; metabolism ; Neuropeptides ; metabolism ; Rats, Sprague-Dawley ; Transfection ; Up-Regulation ; rac1 GTP-Binding Protein ; metabolism