1.Construction of autocatalytic caspase-3 driven by amplified human telomerase reverse transcriptase promoter and its enhanced efficacy of inducing apoptosis in human ovarian carcinoma
Yue SONG ; Keng SHEN ; Chun-Xia HE ;
Chinese Journal of Obstetrics and Gynecology 2000;0(09):-
Objective To construct recombinant adenoviral vector expressing autocatalysis caspase- 3 driven by human telomerase reverse transcriptase promoter amplified by two-step transcription amplification (hTERTp-TSTA),and investigate its antitumor effect in ovarian cancer iri vitro and in vivo.Methods Recombinant adenoviruses expressing autocatalytic caspase-3(rev-caspase-3)driven by hTERTp-TSTA were prepared,which were named as AdHTVP2G5-rev-casp3.AdHT-rev-casp3,Ad-rev-casp3 and AdHTVP2G5- EGEP,which express rev-easpase-3 driven by hTERTp,cytomegalovirus promoter(CMVp)and enhanced green fluorescent protein(EGFP),respectively,were used as controls.Western blot,cell counting kit (CCK-8),flow cytometry(FCM)and TdT-mediated dUTP-biotin nick end labeling(TUNEL)were used to detect the expression of p17,active subunit of caspase-3,and p85,and to measure cell survival rates, apoptotic rates and cell cycle distribution in ovarian cell line AO and normal human umbilical vein endothelial cell line HUVEC,following treatments of AdHTVP2G5-rev-casp3.subcutaneous tumor models and abdominally spread tumor models of human ovarian carcinoma using AO cells in BALB/e nude mice were established.Following treatments of AdHTVP2G5-rev-easp3,western blot was used to detect the expression of active caspase-3 in abdominally spread tumors and liver tissues,respectively,and the mouse survival rates and the volume of tumor nodules were measured,and the serum level of alanine transaminase (ALT)and aspartate transaminase(AST)were analyzed to monitor liver damages and HE staining was used to detect the histopathological changes of various organs.Results The levels of p17 expression in AdHTVP2G5-rev-casp3-treated AO cells were significantly higher than that in Ad-rev-casp3 or AdHT-rev- casp3 treated AO cells,while no expression was observed in AdHTVP2G5-rev-casp3-treated HUVEC.There was strong cell killing of AdHTVP2G5-rev-casp3 of hTERT positive AO cells,but not of the hTERT-negative HUVEC cells.Cell survival rate and apoptotic rate of AO cells treated with AdHTVP2G5-rev-casp3 were 17.8% and 40.2%,respectively,significantly different from that treated with AdHT-rev-casp3(75.2% and 16.1%)at the multiplicity of infection(MOI)of 70(P0.05) .Significant expressions of active caspase-3 were shown in AdHTVP2G5-rev-casp3-treated tumors,whereas no expression was shown in liver.In contrast,both tumors and liver tissues showed active caspase-3 expression following treatments of Ad-rev-casp3.AdHTVP2G5-rev-casp3 and Ad-rev-casp3 prolonged mouse survival[mean survival time of(259?14)d and(213?16)d],when compared with treatment with AdHT- rev-casp3[(177?12)d]and AdHTVP2G5-EGFP[(109?7)d;P
2.Relationship between mitochondrial dysfunction, oxidative stress and diabetic retinopathy
Song, YUE ; Yue-Dong, HU ; Xin-He, WANG ; Lei, CHEN
International Eye Science 2014;(12):2176-2178
As one of the serious complications of diabetes, diabetic retinopathy( DR) has become a main eye disease which causes blindness. The occurrence and development of DR is related to many factors. The pathogenesis is complicated, and the mechanism has not been clear. Early data suggest that the occurrence and development of DR has relations with many factors such as blood sugar level, diabetes duration and the environment. Among the factors, mitochondrial dysfunction and oxidative stress is the important mechanisms of DR and has become research focus in recent years. Consequences of mitochondrial dysfunction within cells include elevation of the rate of reactive oxygen species( ROS) production due to damage of electron transport chain proteins, mitochondrial DNA ( mtDNA ) damage, and loss of metabolic capacity. Clear understanding on the mechanism of mitochondrial functional change under high sugar level and oxidative stress response in the occurrence and development of DR is of great significance on prevention and cure of DR. ln this article, the development of mitochondrial metabolism and oxidative stress of DR is reviewed.
3.Effects of Fragile Mental Retardation Protein on Cerebellar Neuron Development and Migration in Mouse Model
Xiaohong SUN ; Yuxia DONG ; Yu FENG ; Weike SONG ; Yue HE ; Zhiyi HE
Journal of China Medical University 2010;(6):420-421
Objective To investigate the effects of fragile X mental retardation protein(FMRP)on the development and migration of cerebellar neurons in mouse model.Methods Plasmids containing FMRPmutant-EGFP or EGFP were established and transfected into the lateral ventricle of the embryo mouse.Fragile X syndrome(FXS)genotype of the mouse model was identified.Nissl staining and immunofluorescence staining were conducted to assess the changes in neuron development and migration.Results In the experimental group,Nissl staining showed that the deep cerebellar neuclei contracted and divided by white matter,and the non-polarized Purkinje cells retained in internal granular layer;while immunofluorescence staining showed that Tbr2-positive unipolar brush cells changed the migration pathway and accumulated in the ventricular zone.Conclusion Cerebellar neurons showed abnormal formation and migration with the absence of FMRP.
4.Effects of X-rays on CC-chemokine receptor 7 expression in human lung cancer A549 cells
Cuilan WANG ; Qisheng JIANG ; Yue ZOU ; Fengsheng LI ; Wei LI ; Xiujun SONG ; Rui HE ; Lu WANG
Chinese Journal of Radiological Medicine and Protection 2011;31(3):299-302
Objective To study the effects of X-ray radiation on CC-chemokine receptor 7(CCR7) expression in human non-small cell lung cancer (NSCLC) cells.Methods Humanadenocarcinoma cells of the line A549 were cultured and irradiated by X-ray at the absorbed doses of 2,4,6,and 8 Gy respectively by linear accelerator (with the source skin distance of 100 cm and dose rate of 442.89 cGy/min).The relative levels of CCR7 mRNA and protein expression in the A549 cells were respectively detected by real time-PCR and Western blotting 4,12,24,48,and 72 h after radiation.Untreated A549 cells were used as control group.Results The expression levels of CCR7 mRNA and protein in the A549 cells began to increase since 4 h after radiation and then decreased gradually after they reached the peak.The CCR7 mRNA expression levels 72 h after radiation of the 6 and 8 Gy groups were still significantly higher than those of the control group (t = 6.75-7.26,both P < 0.01),and the CCR7 protein expression levels of the 2 and 6 Gy group were still significantly higher than those of the control group(t=11.13-14.17,both P <0.01).Then the CCR7 protein expression levels of the 4 and 8 Gy groups decreased to the control group level 48 and 72 h after radiation respectively.Conclusions The CCR7 mRNA and protein expression levels in the NSCLC cells increase after X-ray irradiation,which may be correlated with the promotion of proliferation and metastasis of NSCLC cells by X-ray irradiation at a certain dose.
5.Clinical observation on 1g/L bromfenac sodium hydrate ophthalmic solution associated with glucocorticoid in high myopia after LASEK
Qing-He, LI ; Yue-Mei, LI ; Xue-Ying, SONG ; Hao, WANG ; Shao-Wen, QI
International Eye Science 2017;17(6):1133-1136
AIM:To observe the efficacy and safety of 1g/L bromfenac sodium hydrate ophthalmic solution in the partial substitution of glucocorticoid after laser subepithelial keratomileusis (LASEK).METHODS: Totally 104 cases (208 eyes) were received LASEK, which were selected and divided into study group and control group.The study group were adopted 1g/L bromfenac sodium hydrate ophthalmic solution combined with tobramycin dexamethasone eye drops and fluorometholone eye drops, the control group were adopted tobramycin dexamethasone eye drops and fluorometholone eye drops.The changes of visual acuity and intraocular pressure of two groups were recorded before and after surgery, and the score of painness and the occurrence of haze were observed after surgery.RESULTS: At postoperative 1, 2, 3d, respectively, the pain score of the study group were 1.70±0.35, 1.25±0.34, 0.82±0.32, the pain score of the control group were 2.30±0.43, 1.68±0.44, 1.12±0.33, the differences were significant (P<0.05).Before and at 2wk, 1, 3mo after surgery, respectively, uncorrected visual acuity of study group were 0.035±0.02, 0.71±0.13, 0.89±0.17, 0.88±0.18, while which of control group were 0.037±0.015, 0.73±0.15, 0.87±0.14, 0.86±0.15 (P>0.05), and the differences were not significant(P>0.05).At preoperative and postoperative 1, 2wk, 1 and 3mo of surgery, respectively, the intraocular pressure of study group were 17.33±1.58, 7.54±1.28, 7.23±1.58, 7.26±1.47, and 7.30±1.36 mmHg;the intraocular pressure of control group were 17.53±1.43,7.57±1.32,7.73±1.55,7.80±1.38,7.86±1.43 mmHg,the differences were not significant between before and at 2wk after surgery(P>0.05), the differences were significant between the two groups at 2wk, 1mo and 3mo (P<0.05).At postoperative 1, 3mo of surgery, the score of haze level of the study group were 91.7% and 93.3%, respectively and which in control group were 92.0% and 92.9%, respectively, and the differences were not significant between the two groups at every time point(P>0.05).CONCLUSION: It is safe and effective that 1g/L bromfenac sodium hydrate ophthalmic solution in the partial substitution of glucocorticoid after laser subepithelial keratomileusis.The patient has a lower intraocular pressure, has similar therapeutic effect as glucocorticoid in vision and antiinflammatory.
6.Study on quality of life of patients with breast cancer after breast conserving surgery
Li XIA ; Baosan HAN ; Jinghua HE ; Hongxia SONG ; Yue ZHAO ; Lin GU
Chinese Journal of Practical Nursing 2010;26(22):1-4
Objective To evaluate the quality of life and influencing factors in patients with breast cancer after breast-conserving surgery, so as to provide theoretical support for preoperative health education and postoperative rehabilitative intervention for these patients. Methods 120 breast conserving surgery (BSC) and 140 modified radical mastectomy (MRM) patients were assessed with Functional Assessment of Cancer Therapy-Breast (FACT-B V4.0) one week after operation. Life quality was evaluated between the two groups, and the influencing factors were also analyzed. Results Postoperative breast cancer patients after breast conserving surgery were with good breast complementary attention, lower emotion state. The other scores of quality of life had no statistical difference. Influencing factors on the quality of life of breast cancer patients were anxiety state, relationship of family members, position of surgery and character type. Conclusions It suggests that nursing specialists should make out corresponding, scientific and reasonable nursing intervention schemes based on the characteristics of different groups of people to ease psychological burden, and elevate the quality of life effectively for postoperative breast conserving surgery patients.
7.Fabrication of a bionic artificial bone scaffold using a room temperature three dimensional printing technique
Kaifeng LIN ; Shu HE ; Yue SONG ; Zheng WANG ; Long BI ; Guoxian PEI
Chinese Journal of Orthopaedic Trauma 2016;18(5):421-427
Objective To prepare a bionic artificial bone scaffold using a room temperature three dimensional (3D) printing technique and evaluate its biocompatibility and bioactivity in vitro.Methods A room temperature 3D printing technique was applied to fabricate 3D bionic artificial bone scaffolds using collagen/hydroxyapatite.The physico-chemical structure,porosity and mechanical strength of the scaffolds were assessed.The extract liquid of scaffolds was cocultured with bone mesenchymal stem cells (BMSCs) to evaluate the toxicity of scaffolds.There were 3 experimental groups:blank control with no scaffolds,printed scaffolds group and non-printed scaffolds group.The condition of BMSCs on the scaffolds was observed via scanning electron microscopy(SEM) and immunostaining.3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay and SEM were applied to monitor the proliferation of BMSCs on the scaffolds.At last,alkaline phosphatase (ALP) activity and mRNA expression levels of osteogenesis-related genes were detected to assess the osteoinductive property of the scaffolds.Results The 3D printed scaffolds fabricated in the present study were characterized by highly interconnected pores which were controllable and even in size.The cross section of the scaffolds presented an irregular honeycomb-like microstructure.The porosity of printed 3D scaffolds (71.14% ± 2.24%) was significantly higher than that of non-printed scaffolds (59.04% ±2.98%) (P < 0.05).The physico-chemical structures of the materials were preserved after printing without additional cytotoxicity.The MTT results at 7 and 14 days revealed that the printed scaffolds had a significantly more cell numbers than the non-printed scaffolds(P < 0.05).SEM showed that the BMSCs adhered well onto the printed scaffolds and proliferated and migrated through the pores.Compared with the blank control,the printed scaffolds showed obviously better osteogenic outcomes.Conclusion The 3D bionic artificial bone scaffolds of collagen/hydroxyapatite manufactured by a room temperature 3D printing technique can provide a good extracellular matrix for BMSCs to proliferate and differentiate.
8.A finite element analysis of torque capabilities of the DamonQ system using the double wire technique
Yanan WANG ; Yang QU ; Song HE ; Siyuan HUANG ; Siwei YANG ; Yue HUANG
Chinese Journal of Tissue Engineering Research 2016;20(33):4919-4925
BACKGROUND:In the orthodontic treatment, double wire technique is commonly used to correct the torque of anterior teeth;however, the underlying mechanical mechanisms remain unclear. OBJECTIVE:To study the torque capabilities of DamonQ system through the three-dimensional finite element method. METHODS:Three-dimensional finite element models of Damon Q brackets with different dimension arch wires were established and were grouped according to arch wire with different dimensions. The torque capabilities of Damon Q system with different dimensions and materials arch wire were analyzed. RESULTS AND CONCLUSION:The Damon Q brackets with single or double wires had the same variation of torque, torque angle curve changes under the condition of different size and material arch wires. Torque moment increased gradually with increasing torque angle. Rectangular-round double wires produced larger torque moment compared with single rectangular wire, whereas double round wires produced smal er torque moment compared with single rectangular wire. The torque moment produced by rectangular-round double wires or by single rectangular wire was approximately 2-8 times or 1-5 times more than that by double round wires. The torque moment that rectangular-round double wires produced was larger than that single wire produced in the DamonQ system. In the early clinical practice, we can use double round wires to control root movement. In the middle-term of the treatment, we can use rectangular-round double wires to control root movement as the mild forces produced by them reduce root absorption. This study lays a foundation for the application of Damon Q bracket with double wire technology in the clinic.
9.Study on Classification Methods for Adjuvant Drugs
Li WANG ; Defang CAI ; Yong CHEN ; Qin HE ; Song ZHANG ; Yuan HOU ; Yue MA
China Pharmacist 2015;18(12):2156-2159
Objective:To define the range and classification of adjuvant drugs. Methods:In order to explore the definition meth-od for the range and classification of adjuvant drugs, the information of definition and classification of adjuvant drugs was obtained by searching PubMed, CNKI, Wanfang database and medicine monographs such as Clinical Medication Notice, New Pharmacology and Martindale:The Complete Drug Reference. Results:The preliminary conclusion on definition, range and classification method for ad-juvant drugs was achieved. Adjuvant drugs were classified into ten categories, so that the adjuvant drugs in our hospital were super-vised. Conclusion:In order to promote the standardized management of clinical application of adjuvant drugs, the range and classifica-tion of adjuvant drugs still need further discussion and standardization.
10.Reparation of human epithelial growth factor receptor 2-targeted polymer ultrasound contrast agents with liquid perfluorocarbons and in vitro experiment
Qinglan KE ; Zhigang WANG ; Zipeng HE ; Yuanyuan YUE ; Pan LI ; Weixiang SONG
Chinese Journal of Ultrasonography 2014;23(1):66-70
Objective To prepare a novel extravascular human epithelial growth factor receptor 2 (HER2)-targeted polylactic-co-glycolic acid (PLGA)-COOH ultrasound contrast agent with liquid perfluorocarbons(PFH-PLGA-tra),and to observe it's general properties and it's effects on ultrasound imaging in vitro.Methods Polymeric nanocapsules of liquid perfluorocarbons were prepared by the single emulsion technique and conjugated with trastuzumab monoclonal antibody by EDC/NHS.The general characteristics were observed.The conjugation was demonstrated by immunofluorescence and the targeting performance of the agent was checked in human MCF-7 cells line in vitro.The encapsulation of liquid fluorocarbons was detected by heating in vitro.The effects on ultrasound imaging were observed in vitro.Results The mean diameter of agents was (261 ± 28)nm.The targeted nanocapsules were positive in immunofluorescence.In the targeted study,it was shown that a number of targeted nanocapsules conjugated with MCF-7 cells in vitro.Droplet-to-bubble transition experiment in vitro showed that there were no nano/microbubbles formed by heating in control groups at 80℃,while there were a lot of nano/microbubbles appeared in nanocapsules formulations at 80 ℃.The agents presented a good ultrasound imaging in vitro.Conclusions The HER2-targeted polymeric nanocapsules of liquid perfluorocarbons are successfully prepared,which may become a novel extravascular targeted ultrasound contrast agents.