4.Construction and identification of a lentiviral vector mediating RNA interference of tyrosine kinase B gene
Mei LI ; Yu ZHAO ; Chao JING ; Deqin GENG ; Baixiang ZHUANG ; Hongbin FAN
Chinese Journal of Behavioral Medicine and Brain Science 2013;(4):370-373
Objective To construct a tyrosine kinase B(TrkB) targeted RNA interference (RNAi) lentiviral vector.Methods Four oligonucleotides targeting rat TrkB gene were synthesized and cloned into lentiviral vector pXZRNAi 1.0 to construct recombinant lentiviral vectors pXZRNAi-shTrkB-1,2,3,4.Neural stem cells prepared from rat hippocampus were infected with these high-titer viruses.Real-time PCR was employed to detect the TrkB mRNA expression and western blot was used to assess the gene silencing efficacy of these recombinants.Results Enzyme digestion and DNA sequencing results demonstrated that these shRNAs were correctly inserted into lentiviral vectors and the four recombinants were constructed successfully with the titer of 8.6 × 105cfu/ml.The infection efficiency of the letivirus on neural stem cells reached 80%.Compared with the uninfection group,the expression levels of TrkB mRNA in neural stem cells decreased significantly after transfected with pXZRNAi-shTrkB-3 and 4((66.7 ± 5.5) % and(76.8 ± 4.9) % respectively,P < 0.05) ; and the protein expression levels were also significantly decreased ((68.5 ± 4.3)% and (78.2 ± 5.1)% respectively,P < 0.05).Conclusion The lentiviral vectors for TrkB have been successfully constructed with high yield of lentivirus,which provides versatile method for assessing gene function in neural stem cells.
5.Preparation of Polydiallyl Dimethyl Ammonium Chloride Modified Reduced Graphene Oxide/Zinc Oxide Composite Materials and Its Application in Detection of Uric Acid
Mei Yong FAN ; Mei Ji ZHANG ; Chao MI ; Ling Yu CAI ; Ping Li ZHANG ; Bin Hai ZHU
Chinese Journal of Analytical Chemistry 2018;46(1):33-38
By using Zinc nitrate as precursor and hydraZine hydrate as reducing agent, polydiallyl dimethyl ammonium chloride modified reduced graphene oxide/Zinc oxide composite materials ( PDDA-rGO/ZnO) were prepared by simultaneous reduction of graphene oxide ( GO) and Zinc nitrate.The composite materials were characteriZed by Fourier transform infrared ( FTIR ) spectroscopy, X-ray diffractometer ( XRD ) and transmission electron microscopy ( TEM) , and their electrochemical catalytic activity for uric acid was studied by cyclic voltammetry ( CV ) and linear sweep voltammetry ( LSV ) measurements.The result showed that PDDA-rGO/ZnO modified glassy carbon electrode prepared here was sensitive, reproducible and stable, and had significant electrocatalytic activity for UA.When using linear sweep voltammetry for detection of UA, the responses of modified electrode were linear with UA concentration in the ranges of 0.02-0.1 mmol/L and 0.1-1.0 mmol/L respectively, with detection limit of 15.9 nmol/L (S/N=3).
6.Evidence for determining the safe surgical margin for pleomorphic adenoma of parotid gland.
Yi-quan XU ; Chao LI ; Jin-chuan FAN ; Bing ZHANG ; Jian-chao CHEN ; Zhao-hui WANG ; Ying WANG ; Sheng QIN ; Xiao-yan QING ; Jun-feng WAN ; Lun LI ; Mei TANG ; Hong YANG ; Bo LIANG
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2012;47(2):137-141
OBJECTIVETo compare the treatment outcomes, complications and histopathologic features between conventional parotidectomy and functional regional parotidectomy in the treatment for pleomorphic adenoma of parotid gland and to provide clinical, and pathological evidence for determining the safe surgical margin.
METHODSOf 109 patients, 60 patients received conventional parotidectomy and 49 patients received functional regional parotidectomy. The rates of tumor recurrence and complications were compared between the groups of patients.
RESULTSThere was no significant difference in the incidence of tumor recurrence, the facial paralysis and sialosyrinx between two groups. The rates of Frey's syndrome, numbness of auricular region, and facial asymmetry were 30.0%, 61.7%, and 38.3% in the patients with conventional parotidectomy respectively, while the rates were 6.1%, 30.6%, and 8.2% in the patients with functional regional parotidectomy, with significant statistically difference, respectively (P < 0.05). Of 109 patients, 33 with incomplete capsule, 29 with capsule penetration, 25 with pseudopodia, and 13 with satellite nodules. There was no significant difference in the depth of tumor infiltration between two groups of patients. For the tumor smaller than 2 cm, the depth of infiltration in conventional group was from 0.061 to 1.122 mm, functional group was from 0.442 to 3.127 mm (Z = -1.093, P = 0.057); for the tumors between 2 - 4 cm, the depth in conventional group was from 0.081 to 7.908 mm, functional group was from 0.082 to 6.632 mm (Z = -0.214, P = 0.831); for the tumor larger than 4 cm, the depth of infiltration was from 0.340 to 8.476 mm.
CONCLUSIONSCompared with conventional parotidectomy, functional regional parotidectomy has good outcomes and less complications. The surgical margins of pleomorphic adenomas of the parotid gland should be determined by the size of tumor. The 1 cm-surgical margins are safe for the tumors less than 4 cm, and the tumors more than 4 cm should be treated with superficial parotidectomy.
Adenoma, Pleomorphic ; pathology ; surgery ; Adolescent ; Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; Parotid Neoplasms ; pathology ; surgery ; Retrospective Studies ; Treatment Outcome ; Young Adult
7.Clinical features and prognosis of 188 Chinese patients with apical hypertrophic cardiomyopathy.
Yong-kang TAO ; Li-rong YAN ; Yi-shi LI ; Zhi-min XU ; Guo-qiang ZHANG ; Li-mei ZU ; Hai-yan DU ; Chao-mei FAN
Chinese Journal of Cardiology 2011;39(2):106-109
OBJECTIVETo analyze the clinical characteristics and prognosis of Chinese patients with apical hypertrophic cardiomyopathy (AHCM).
METHODSA total of 188 patients with AHCM diagnosed at Fuwai Hospital were included in this retrospective study. Clinical characteristics, mortality and cardiovascular morbidity were analyzed. A multiple logistic regression was performed to adjust for potential confounding factors.
RESULTSMales predominated with a number of 139 (73.9%) in this cohort. Patient's age ranged from 15 to 81 (51.9 ± 12.6) years. There were 120 patients (63.8%) with "pure" type and 68 patients (36.2%) with "mixed" type of AHCM, 171 patients were followed up for (5.0 ± 3.0) years, cardiovascular mortality was 1.2%, 28 patients (16.4%) experienced one or more cardiovascular events.
CONCLUSIONThe prevalence of AHCM is high in Chinese HCM patients, pure type AHCM is more common, and AHCM patients have a benign clinical course.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Cardiomyopathy, Hypertrophic ; diagnosis ; epidemiology ; Female ; Humans ; Male ; Middle Aged ; Prognosis ; Retrospective Studies ; Young Adult
8.Mitofusin2 decreases intracellular cholesterol of oxidized LDL-induced foam cells from rat vascular smooth muscle cells.
Chao HE ; Ying CHEN ; Chun LIU ; Ming CAO ; Yu-jin FAN ; Xiao-mei GUO
Journal of Huazhong University of Science and Technology (Medical Sciences) 2013;33(2):212-218
Mitofusin2 (Mfn2) plays a pivotal role in the proliferation and apoptosis of vascular smooth muscle cells (VSMCs). The purpose of this study was to investigate the effects of Mfn2 on the trafficking of intracellular cholesterol in the foam cells derived from rat VSMCs (rVSMCs) and also to investigate the effects of Mfn2 on the expression of adenosine triphosphate-binding cassette subfamily A member 1 (ABCA1), adenosine triphosphate-binding cassette subfamily G member 1 (ABCG1) and peroxisome proliferator-activated receptor gamma (PPARγ). The rVSMCs were co-cultured with oxidized low density lipoprotein (LDL, 80 μg/mL) to produce foam cells and cholesterol accumulation in cells. Before oxidized LDL treatment, different titers (20, 40 and 60 pfu/cell) of recombinant adenovirus containing Mfn2 gene (Adv-Mfn2) were added into the culture medium for 24 h to transfect the Mfn2 gene into the rVSMCs. Then the cells were harvested for analyses. The protein expression of Mfn2 was significantly higher in Adv-Mfn2-transfected group than in untransfected group (P<0.05), and the expression levels significantly increased when the titer of Adv-Mfn2 increased (P<0.05). At 24 or 48 h after oxidized LDL treatment, rVSMCs became irregular and their nuclei became larger, and their plasma abounded with red lipid droplets. However, the number of red lipid droplets was significantly decreased in Adv-Mfn2-transfected group as compared with untransfected group. At 48 h after oxidized LDL treatment, the intracellular cholesterol in rVSMCs was significantly increased (P<0.05), but it was significantly decreased in Adv-Mfn2-transfected group as compared with untransfected group (P<0.05), and it also significantly decreased when the titer of Adv-Mfn2 increased (P<0.05). The mRNA and protein expression levels of ABCA1 and ABCG1 were significantly increased in Adv-Mfn2-transfected group as compared with untransfected group (P<0.05). Though the mRNA and protein expression levels of PPARγ was not significantly increased (P>0.05), the phosporylation levels of PPARγ were significantly decreased in Adv-Mfn2-transfected group as compared with untransfected group (P<0.05). These results suggest that the transfection of Adv-Mfn2 can significantly reduce intracellular cholesterol in oxidized LDL-induced rVSMCs possibly by decreasing PPARγ phosporylation and then increasing protein expression levels of ABCA1 and ABCG1, which may be helpful to suppress the formation of foam cells.
ATP Binding Cassette Transporter 1
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metabolism
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ATP Binding Cassette Transporter, Sub-Family G, Member 1
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ATP-Binding Cassette Transporters
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metabolism
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Animals
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Cell Differentiation
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physiology
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Cells, Cultured
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Cholesterol
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metabolism
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Foam Cells
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cytology
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metabolism
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Intracellular Fluid
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metabolism
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Lipoproteins, LDL
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metabolism
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Membrane Proteins
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genetics
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metabolism
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Mitochondrial Proteins
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genetics
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metabolism
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Muscle, Smooth, Vascular
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cytology
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metabolism
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Oxidation-Reduction
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PPAR gamma
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metabolism
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Rats
9.Impact of application time of aspirin and clopidogrel on platelet aggregation in patients with acute coronary syndrome.
Zhen LI ; Fan LIU ; Wei CUI ; Rui-qin XIE ; Xiu-chun YANG ; Jing-chao LU ; Hong-mei ZHENG ; Xiao-juan REN ; Jing LIU
Chinese Journal of Cardiology 2010;38(4):311-314
OBJECTIVETo observe the impact of various application time of aspirin and clopidogrel on the circadian rhythm changes of platelet aggregation in patients with acute coronary syndrome.
METHODSPatients with acute coronary syndrome were divided into day-time (8:00) and night-time (20:00) medication group (n = 15 each). After plasma concentration reached steady state, platelet aggregation was assessed at 5 time points within 24 hours with a mobile four-channel whole blood impedance aggregometer. The platelet aggregation was induced by ADP and arachidonic acid. Thereafter, the two groups were exchanged and platelet aggregation was assessed in the same way post plasma steady state.
RESULTSArachidonic acid-induced platelet aggregation was the highest at 10:00 Am [(7.96 +/- 3.64) ohm] and the lowest at 0:00 [(6.12 +/- 3.29) ohm, P > 0.05] in day-time group. Platelet aggregation was the highest at 20:00 [(9.40 +/- 5.39) ohm] and the lowest at 10:00 [(5.46 +/- 3.93) ohm], P < 0.05). ADP-induced platelet aggregation was the highest at 10:00 and the lowest at 16:00 in day-time group (P > 0.05) and was the highest at 20:00 and the lowest at 10:00 in night-time group (P > 0.05). Platelet aggregation induced by two inducers was significantly higher at 10:00 in day-time group compared to values in night-time group (all P < 0.05).
CONCLUSIONTaking aspirin and clopidogrel at 20:00 was superior to taking the same medications at 8:00 for inhibiting peak platelet aggregation in the morning.
Acute Coronary Syndrome ; drug therapy ; Adult ; Aged ; Aspirin ; administration & dosage ; therapeutic use ; Circadian Rhythm ; Humans ; Male ; Middle Aged ; Platelet Aggregation ; drug effects ; Platelet Aggregation Inhibitors ; administration & dosage ; therapeutic use ; Platelet Function Tests ; Ticlopidine ; administration & dosage ; analogs & derivatives ; therapeutic use ; Time Factors
10.3D image analysis of nasal orbit fossa through a laser scanner: a new method and application of 3D image.
Xiang-Dong QI ; Jia-Zeng QIN ; Wei-Dong ZHAO ; Ji-Hong FAN ; Jian-Tie LI ; Mei-Chao ZHANG ; Wen-Hua HUANG ; Shi-Zhen ZHONG
Chinese Journal of Plastic Surgery 2004;20(4):252-255
OBJECTIVEThis study was to introduce the method for obtaining accurate 3D data of soft tissues using a 3D scanner under non-contact condition and the standard 3D measurement of the nasal orbit fossa for plastic surgery.
METHODSA 3D laser scanner and the Geomagic software were used to obtain the standard facial contour of 30 Chinese people. The nasal orbit fossa, as the feature of beauty, was measured and the data were analyzed.
RESULTS3D measurement exhibited the three-dimensional facial shape at every meaningful angle, with the advantages of high precision of 0.01 mm. We determined the lowest point and described the 3D feature of the nasal orbit fossa.
CONCLUSIONSThis method can illustrate the relation of the nasal orbit fossa and the surrounding structure. It is a new approach to facilitate preoperative plans, operation simulation and postoperative evaluation.
Adult ; Female ; Humans ; Image Processing, Computer-Assisted ; Imaging, Three-Dimensional ; instrumentation ; methods ; Lasers ; Male ; Nose ; anatomy & histology ; Orbit ; anatomy & histology