2.5-Aza-2 ’-deoxycytidine and 4-phenylbutyric acid exert a cooperative effect on the expression of miR-196 b in chronic myeloid leukemia cells
Yue LIU ; Chun SHUAI ; Jiesheng LI ; Hong YIN ; Yanbin SONG ; Wenli MA
Acta Anatomica Sinica 2014;(4):521-524
Objective To study if 5-Aza-2’-deoxycytidine along or together with 4-phenylbutyric acid could affect miR-196b expression levels in chronic myeloid leukemia cells .Methods K562 cells were treated with DNA methylation inhibitor 5-Aza-2’-deoxycytidine, histone deacetylase inhibitors 4-phenylbutyric acid separately and the combined treatment with both of them, then expression levels of miR-196b were detected using Real-time PCR.Results The half inhibition concentration of 4-phenylbutyric acid was 1.58mmol/L.Comparing with the expression level of miR-196b in normal human bone marrow cells, the expression levels of miR-196b were significantly lower in Aza group , PBA group and negative control cells and nearly consistent among three groups , and as high as normal cells in combined treatment group . Conclusion The expression level of miR-196b in K562 cells could not return to normal treated with 5-Aza-2 ’-deoxycytidine or 4-phenylbutyric acid separately , while could restore normal when treated with both agents , indicating that miR-196b expression level in K562 cells is related with both DNA methylation and histone acetylation .
3.Application of Cell Counting Kit-8 in detecting the growth inhibiting effect of 5-Aza-2 ’-deoxycytidine on chronic myeloid leukemia cells
Yue LIU ; Chun SHUAI ; Jiesheng LI ; Hong YIN ; Yanbin SONG ; Wenli MA
Acta Anatomica Sinica 2014;(4):582-584
Objective To study the application of Cell Counting Kit-8(CCK-8) in detecting the growth inhibiting effect of 5-Aza-2’-deoxycytidine on chronic myeloid leukemia cell .Methods The proliferation of K562 cells was detected by CCK-8 with different concentrations of 5-Aza-2 ’-deoxycytidine and the cell cycle and apoptosis of K 562 cells were detected after K562 treated by 50% inhibitory concentration of 5-Aza-2 ’-deoxycytidine .Results The 50% inhibitory concentration of 5-Aza-2’-deoxycytidine was 15.55nmol/L, after treated with this concentration , K562 cells showed that G2 phase arrest occurred , proliferation inhibited and apoptosis peaks appeared .Conclusion Inhibition of proliferation of K562 cells with different concentrations of 5-Aza-2’-deoxycytidine varied in a dose-dependent relationship , and 5-Aza-2’-deoxycytidine could promote apoptosis of K 562 cells.CCK-8 can be used in detecting the effect of 5-Aza-2’-deoxycytidine on chronic myeloid leukemia cells .
4.Effects of siRNA interference combined with hyperbaric oxygen treatment on cerebral edema and apoptosis in the brain tissue of rats after hemorrhage
Qiang PAN ; Lin ZHU ; Shuai ZHANG ; Puxian LI ; Xingtao DIAO ; Chunyu SONG ; Yong GAO ; Feng SI ; Qiang LI ; Xiao YUE
Chinese Journal of Geriatrics 2017;36(9):1010-1014
Objective To examine the effects and mechanisms of siRNA targeting aquaporin 4 (AQP 4) in combination with hyperbaric oxygen therapy(HBO) on cerebral edema and apoptosis in the brain tissue of rats after hemorrhage.Methods Rats were randomly divided into four groups,the control group,the hyperbaric oxygen group,the AQP-4 siRNA group and the combination therapy group (24 rats).Thrombin Ⅶ was injected into the caudate nucleus to establish the hemorrhage model.Construction of siRNA targeting aquaporin 4 was conducted.The mRNA expression of AQP-4 was detected by RT-PCR at day 3.Changes in brain moisture and blood-brain barrier perme ability were measured by a wet/dry weight method and Evans blue fluorometry.The nerve cell apoptosis rate was analyzed by Annexin V andTdT-mediated dUTP-biotin nick end labeling (TUNEL).The expression of proteins including AQP-4,MMP-2,MMP-9,Bcl-2 and caspase-3 was detected by Western Blotting.All the animals were given a score for their nerve function at day 3.Results AQP-4 siRNA treatment obtained better effects than HBO in decreasing the brain edema leveland silencing AQP-4 mRNA(P<0.05)while,the combination therapy group achieved the best results(P< 0.05).Compared with the control group,the percentage of apoptotic cells decreased in all the three treatment groups,with the most marked decrease observed in the combination treatment group(4.24± 0.04)%(F=13.76,P=0.001).The expression of AQP-4,MMP-2,MMP-9 and caspase-3 was lower (P<0.05) and the expression of Bcl-2 was higher(P<0.01)in the combination treatment group than in the other three groups.Compared with the control group,all the other three groups received better scores on nerve function defect evaluation at day 3 after hemorrhage(P<0.05),with the combination treatment group again achieving the most favorable score (4.7 ± 1.1) (F=7.21,P =0.013).Conclusions Targeted siRNA interference combined with hyperbaric oxygen can effectively reduce cerebral edema after cerebral hemorrhage,inhibit neuronal apoptosis and promote neuron function recovery.The underlying mechanisms may be related to down-regulation of AQP-4,MMP 2,MMP-9 and caspase-3 expression and up-regulation of Bcl-2 expression.
5.Operative cooperation in cochlear implantation with intraoperative CT scanning
Mei-Ning YUAN ; Yue-Shuai SONG ; Qiu-Ling ZHANG ; Li-Xia LI ; Yong-Yi YUAN
Chinese Journal of Modern Nursing 2013;19(2):145-148
Objective To explore the operative cooperation in cochlear implantation with intra-operative CT scanning.Methods Ten cases of cochlear implantation with intraoperative CT scanning between April 2010 and April 2011 were selected and summarized the characters of operations.Results The electrode arrays were placed adequately in all ten cases.The duration of operation was (2.3 ± 0.5)h,the times needed for scanning during operation was (1.4 ± 0.7) h,the duration of intraoperative CT scanning was (13.5 ± 6.7) min.Conclusions The complex cochlear implantation with intraoperative CT scanning can be smoothly finished with proper preoperative and scientific and reasonable cooperation.
6.Long-term results of surgical repair of ruptured sinus of aortic sinus aneurysm.
Ran DONG ; Bao-tian CHEN ; Xu MENG ; Tao-shuai LIU ; Yue SONG ; Ju-bing ZHENG ; Yi-hua HE
Chinese Journal of Surgery 2008;46(24):1913-1915
OBJECTIVETo summarize the clinic characteristics and effect of surgical repair of ruptured aortic sinus aneurysm.
METHODSFrom September 1997 to September 2007, 43 patients with ruptured aortic sinus aneurysm underwent surgical procedures. There were 32 male and 11 female patients. The age ranged from 11 to 50 years old with a mean of (29.0 +/- 11.5) years old. The origins of rupture were the right coronary sinus in 34 patients and the noncoronary sinus in 9 patients. The aneurysms ruptured into the right ventricle in 30 patients, the right atrium in 8 patients, the right ventricle and right atrium in 3 patients, and the ventricular septum and then the right ventricle in 2 patients. Associated cardiac anomalies included ventricular septal defect in 26 patients, aortic regurgitation in 15 patients, infectious endocarditis in 8 patients, tricuspid regurgitation in 6 patients, atrial septum defect in 4 patients, mitral valve regurgitation in 2 patients, patent ductus arteriosus in 2 patients, and pulmonary valve vegetation in 1 patient. All the patients underwent the repair of ruptured aortic sinus aneurysm and correction of associated anomalies with cardiopulmonary bypass.
RESULTSThere were no deaths after the operation and during the follow-up. The complications, including acute heart failure and III atrioventricular block, occurred in 5 patients. Follow-up was 6 to 120 months with a mean of (68.0 +/- 17.7) months. Two patients underwent reoperation for aortic valve replacement at the 6(th) and 8(th) year after the first operation. There were 2 patients which the aortic regurgitation deteriorated from grade I to II.
CONCLUSIONSRepair of ruptured aortic sinus aneurysm presents a satisfactory result. Aggressive treatment in early time, prevention of post-operative complications and long-term follow-up are recommended in the treatment for patients of ruptured aortic sinus aneurysm with infectious endocarditis and aortic regurgitation.
Adolescent ; Adult ; Aortic Rupture ; surgery ; Child ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Sinus of Valsalva ; surgery ; Treatment Outcome
7.Effect and mechanism of hyperbaric oxygen combined with aquaporin-4 gene silencing on cognitive dysfunction in rats with traumatic brain injury
Qiang PAN ; Lin ZHU ; Yong GAO ; Chunyu SONG ; Xingtao DIAO ; Shuai ZHANG ; Xiao YUE
Chinese Journal of Behavioral Medicine and Brain Science 2021;30(8):686-693
Objective:To investigate the effect of hyperbaric oxygen combined with RNA interference (RNAi) technology targeting aquaporin-4 (AQP-4) on improving cognitive function in rats with traumatic brain injury (TBI), and to explore its mechanism.Methods:Totally 112 adult male SD rats were randomly divided into four groups: control group, hyperbaric oxygen(HBO) group, AQP-4 RNAi group and combined treatment group, with 28 rats in each group.The TBI model of rat was established by hydraulic percussion and siRNA targeting aquaporin 4 was constructed. Rats were given corresponding intervention according to their groups.Then the modified neurological severity scores(mNSS)was evaluated on the 7th day and 21th day after operation. Morris water maze test was carried out from the 21st day to 25th day after operation and the percentage of target quadrant and daily escape latency were recorded.The changes of the brain permeability of blood-brain barrier and moisture in brain tissues were measured by Evans blue fluorometry and a wet-dry-weighing technique respectively. The protein expression levels of AQP-4, Caspase-3, Bcl-2, MMP-2 and MMP-9 were detected by Western blot method. The mRNA expression of AQP-4 in TBI brain tissue was measured by RT-PCR method, and the apoptosis rate of TBI brain cells was detected by TUNEL and AnnexinV methods on the 7th day after operation. SPSS 23.0 and Graphpad Prism 7.0 softwares were used for data analysis.One-way ANOVA was used for inter group comparison.Repeated measurement ANOVA was used for Morris results, and the LSD- t test was used for pairwise comparisons. Results:The results of mNSS showed that there were significant differences among the groups on the 7th day and 21st day after operation ( F=4.89, 7.59, both P<0.05). The scores of each treatment group were lower than that of the control group, and the effect of the combined treatment group was the best (7th day: t=3.98, -7.75, both P<0.05; 21st day: t=47.82, 7.94, both P<0.05). The results of Morris water maze test showed that the time and group interaction of rats in the target quadrant residence time and escape latency were not statistically significant( F=1.83, 8.42, both P>0.05). The escape latency and the percentage of stay in the target quadrant in the combined treatment group were better than those in other groups on the 24th and 25th day after operation (all P<0.05). Evans blue staining showed that the contents of Evans blue in AQP-4 RNAi group, hyperbaric oxygen group and combined treatment group were lower than that in the control group(all P<0.05), and that in the combined treatment group was the lowest( t=6.19, P<0.05). The results of dry-wet specific gravity method showed that the water content of brain tissue in the combined treatment group((68.15±1.52)%) was the lowest, and that in the AQP-4 RNAi group((76.71±1.06)%) was lower than that in the HBO group ((80.23±1.43)%)( t=4.38, P<0.05). The results of Western blot showed that the protein levels of AQP-4, Caspase-3, MMP-2 and MMP-9 in the combined treatment group were significantly lower than those in other groups(all P<0.05), while the expression of Bcl-2 was increased in the combined treatment group( P<0.05). RT-PCR results (gray value ratio) showed that AQP-4 mRNA levels in AQP-4 RNAi group(0.61±0.21), HBO group (0.83±0.12), combined treatment group(0.22±0.05) and CON group (1.31 ± 0.25) were significantly different( F=175.05, P<0.05), while the AQP-4 mRNA levels decreased in AQP-4 RNAi group which was better than that in hyperbaric oxygen group ( t=5.25, P<0.05). The decrease was the most obvious in the combined treatment group ( t=58.94, P<0.05). The results of TUNEL and AnnexinV showed that the treatment groups were more effective than the control group in inhibiting neuronal apoptosis, especially in the combined treatment group ( P<0.01). Conclusion:The combination of targeted AQP-4 RNAi and hyperbaric oxgen can effectively promote the recovery of neurological and cognitive function, and the mechanism may be related to protecting the integrity of blood-brain barrier, alleviating brain edema and inhibiting apoptosis of nerve cells after TBI.
8.Evaluation of antinociceptive and anti-inflammatory effects of aqueous extract of Armadillidium vulgare Latreille.
Shen-Shen GUO ; Meng-Yue REN ; Shuai SONG ; Ping WEI ; Jia-Bo LUO
Chinese journal of integrative medicine 2017;23(2):138-145
OBJECTIVETo assess the antinociceptive and anti-inflammatory properties of the aqueous extract of Armadillidium vulgare (AV).
METHODSThe antinociceptive effect of AV (400, 600 and 800 mg/kg) was investigated in mice using the acetic acid-induced writhing, formalin-induced nociceptive, and hot plate tests. Phlogogen-induced paw edema using carrageenan, dextran, or compound 48/80 as phlogogen was used as inflammatory models to evaluate AV's anti-inflammatory effect. Additionally, the bioactive substances glucosamine (GLcN) and taurine in AV were determined using high-performance liquid chromatography.
RESULTSOral treatment of the mice with AV (600 and 800 mg/kg) significantly reduced the number of writhes in the acetic acid-induced writhing test (P<0.01) but not the hot plate test (P>0.05). All doses tested significantly inhibited paw-withdrawal during the second phase of the formalin-induced nociceptive model (P<0.01). AV demonstrated a strong anti-inflammatory effect in all those inflammatory models (P<0.05).
CONCLUSIONSAV has antinociceptive and anti-inflammatory effects, providing scientific evidence of the efficacy of its traditional use in pain treatment. Furthermore, GLcN and taurine contribute, at least in part, to the anti-inflammatory activity of AV.
Analgesics ; pharmacology ; Animals ; Anti-Inflammatory Agents ; pharmacology ; Edema ; drug therapy ; Female ; Inflammation ; drug therapy ; Isopoda ; chemistry ; Male ; Mice ; Pain ; drug therapy ; Pain Measurement ; Phytotherapy ; Plant Extracts ; chemistry ; pharmacology ; Water ; chemistry
9.Optimization of β-globin Stable Expression Using the Third Generation Lentiviral Vector for β-thalassemia Therapy.
Zhen YU ; Shuai TONG ; Yue BAI ; Xiao-Song ZHONG
Journal of Experimental Hematology 2022;30(3):844-850
OBJECTIVE:
To provide a research basis for a safe and effective cell therapy for β-thalassemia through optimization of HS4 region of the third generation lentiviral vector for stable expression of β-globin.
METHODS:
The human β-globin HS4 region in the third generation lentiviral expression vector was optimized to construct the lenti-HBB, and the transcription and translation of β-globin gene were analyzed by RT-PCR and Western blot after the transduction of lenti-HBB in MEL cell line. Furthermore, the erythroid differentiation of CD34+ cells which were transduced lentiviral virus carrying human β-globin from normal human umbilical cord blood cells and peripheral blood cells of patients with β-thalassemia major were confirmed by colony formation assay, cell smear assay and flow cytometry. The safety and effectiveness of the optimized lenti-HBB were verified by NSG mouse in vivo test.
RESULTS:
The human β-globin was expressed stably in the MEL cells, and CD34+ cells from health umbilical cord blood as well as PBMC from patient with β-thalassemia major transduced with lenti-HBB could be differentiated to mature red blood cells. The β-globin expression and differentiation in CD34+ cells were demonstrated successfully in the NSG mouse for about 35 months after post-transplant.
CONCLUSION
Stable β-globin expression through the optimization of HS4 from CD34+ in the third generation lentiviral vector is safe and effective for patients with severe β-thalassemia and other β-globin abnormal diseases.
Animals
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Genetic Therapy
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Genetic Vectors
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Humans
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Lentivirus/genetics*
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Leukocytes, Mononuclear
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Mice
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beta-Globins/genetics*
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beta-Thalassemia/therapy*
10.Evaluation of life cycle management system on patients'prognosis after transcatheter aortic valve replacement
Ruo-Yun LIU ; Ran LIU ; Mei-Fang DAI ; Yue-Miao JIAO ; Yang LI ; San-Shuai CHANG ; Ye XU ; Zhi-Nan LU ; Li ZHAO ; Cheng-Qian YIN ; Guang-Yuan SONG
Chinese Journal of Interventional Cardiology 2024;32(6):311-316
Objective With the widespread of transcatheter aortic valve replacement(TAVR)in patients with severe symptomatic aortic stenosis(AS),the life-cycle management has become a major determinant of prognosis.Methods A total of 408 AS patients who underwent successfully TAVR from June 2021 to August 2023 were consecutively enrolled in Hospital Valve Intervention Center.Patients were assigned to the Usual Care(UC)group between June 2021 and October 2022,while patients were assigned to the Heart Multi-parameter Monitoring(HMM)group between November 2022 and August 2023.The primary endpoint was defined as composite endpoint within 6 months post-TAVR,including all-cause death,cardiovascular death,stroke/transient ischemic attack,conduction block,myocardial infarction,heart failure rehospitalization,and major bleeding events.Secondary endpoints were the time interval(in hours)from event occurrence to medical consultation or advice and patient satisfaction.Statistical analysis was performed using Kaplan-Meier and multivariable Cox proportional hazards models.Results The incidence of primary endpoint in HMM group was significantly lower than that in UC group(8.9%vs.17.7%,P=0.016),the driving event was the rate of diagnosis and recognition of conduction block.The average time intervals from event occurrence to receiving medical advice were 3.02 h in HHM group vs.97.09 h in UC group(P<0.001).Using cardiac monitoring devices and smart healthcare platforms provided significant improving in patients long-term management(HR 0.439,95%CI 0.244-0.790,P=0.006).Conclusions The utilization of cardiac monitoring devices and smart healthcare platforms effectively alerted clinical events and improved postoperative quality of life during long-term management post TAVR.