Objective To obtain an easy and high efficient method for gene transfer to cochlear hair cells ,by comparing three mediating green fluorescent protein (GFP) methods (electroporation ,adenovector and lentivirus vector) .Methods Cochlear sensory epithelium was dissected from anaesthetized P2 mice .Sensory epithelia were transferred onto poly -L -lysine treated cover slides and cultured overnight .Gene transfer was performed by elec‐troporation in medium containing pGPHI/GFP/Neo plasmid or by incubation with diluted recombined adenovirus/lentivirus vector .After 48 hours ,green fluorescence was checked under fluorescence microscope .To confirm the ef‐ficiency of exogenous gene transfer ,real-time PCR was performed using specific primers .Results The transfec‐tion efficiencies of electroporation and lentivirus vector mediated gene transfer were very low .Both immunofluores‐cence and real - time PCR results showed that the transfection efficiency of adenovirus mediated GFP and Bmi1 transfer were relatively higher .The proportion of GFP positive cells in outer hair cells and inner hair cells of middle turn were 90 .0% ± 4 .1% and 5% ± 0 .4% ,respectively .Conclusion Adenovector is more efficient for exogenous gene transfer to cochlear hair cells ,thus adenovector is a good carrier for gene transfer to cochlear hair cells .

Bronchi ; cytology ; Cell Culture Techniques ; Cells, Cultured ; Humans ; Myocytes, Smooth Muscle ; cytology

UNLABELLEDTo investigate the effect of immune response mediated by the T cells stimulated with the specific antigen (oxidized low-density lipoprotein, oxLDL) on plaque stability in coronary heart disease.

METHODSThis study involved 20 patients with acute myocardial infarction (AMI), 34 with unstable angina pectoris (UAP), 27 with stable angina pectoris (SAP) and 22 healthy control subjects. With MTS/PMS colorimetric assay, the T cells from all the subjects were tested for proliferative response to stimulation by 5 microg/ml oxLDL and 5 microg/ml low-density lipoprotein (LDL). Interferon-gamma (IFN-gamma) concentration in the proliferative response of the T cells was measured with enzyme-linked immunosorbent assay (ELISA).

RESULTSThe proliferative response of the T cells elicited by 5 microg/ml oxLDL stimulation was significantly higher in the AMI and UAP groups than in the SAP and control groups (P<0.05). Similarly, IFN-gamma concentration in the proliferative response of the T cells to 5 microg/ml oxLDL stimulation was significantly higher in the former two groups (P<0.05). In the AMI and UAP groups, 5 microg

CONCLUSIONThe immune response mediated by the T cells to specific antigen stimulation, especially the immune response mediated by T helper type 1 (Th1) cells secreting IFN-gamma, may play an important role in the plaque instability and the occurrence of acute coronary syndrome.

Aged ; Coronary Disease ; immunology ; Female ; Humans ; Interferon-gamma ; secretion ; Lipoproteins, LDL ; immunology ; Male ; Middle Aged ; T-Lymphocytes ; immunology ; metabolism ; T-Lymphocytes, Helper-Inducer ; immunology ; metabolism

0.05),but a significant difference at weeks 4,8,and 12 between two groups(P

Objective To compare the difference of the protein about the patient of hepatitis B who received adefovir dipivoxil(ADV)therapy,and seek the useful biomarker of effective therapy.Methods We used the two-dimensional gel electrophoresis technology to examine HBV infected serum samples aiming at searching protein's alteration after ADV therapy.Results After 1 year's treatment,haptoglobin, haptoglobin 2-alpha raised and alpha-l-antitrypsin precursor,Factor B,Chain B,transthyretin,glutathione peroxidase,alpha-2-HS-glycoprotein,retina]binding protein,retinol-binding protein precursor, apolipoprotein,apolipoprotein A-I precursor fell in viral response patients.Transthyretin raised and leucine- rich alpha-2-glyeoprotein,haptoglobin,alpha-2-actin,apolipoprotein A-I precursor fell in none viral response patients.To compare two groups:apolipoprotein A-I have the same change and haptoglobin, transthyretin have the opposite change.Conclusion Proteomics study can find the alteration of protein during the ADV treatment,and is helpful to searching the predictable biomarker to ADV.

To develop a reverse dot blot assay for rapid detection of HBV genotypes.Specific oligonucleotides probes were desighed and immobilized on nylon membranes.The DNA sample to be tested was PCR-amplified with DIG labeling primers and then hybridized with the immobilized probes.This procedure for detecting HBV genotypes was simple,rapid and specificity.30 specimens in Chongqing area were collected and detected by this method,and results were evaluated using direct sequencing.Results showed that: This new method was applicable to precise detection HBV genotypes for specimen with copies up to 103,and the HBV genotyping results showed that genotype B was the predominant genotype in Chongqing area.

Objective To investigate the curative effect of acute severe brain injury complicated ARDS, Methods 31 patients who had acute severe brain injury complicated ARDS were divided into two groups:A group was early discovery of ARDS and given treatment.B group was late discovery of ARDS and treated late.Then the curative effects were compared.Results A group was significantly higher than B group in blood gas analysis(P

Aim To investigate the effects of salidroside on NeuN and Egrs in the ischemic side of middle cerebral artery occlusion (MCAO) rats by inhibiting complement C3.Methods The rats were subjected to MCAO with suture-occluded method,and the neurologic injury was evaluated.The rats underwent l h ischemia/reperfusion with 1 d and 2d salidroside treatment,and the expressions of NeuN,Egr4,C3 and C1 were tested.Male Sprague Dawley rats were separately injected ventricle C3aR inhibitor and artificial cerebrospinal fluid with the help of ventricle stereotaxic apparatus.Thirty min later,the models of MCAO were finished with 1h reperfusion,drug administration and intracerebroventricular injection for 2d.The expressions of NeuN,Egr4,C3 were detected.Results Compared with models of MCAO,the expression of C3 in MCAO rats treated with salidroside 1 d and 2d decreased significantly,and the expression of NeuN increased markedly.Salidroside had no apparent effect on Egr4 and C1 of administration of 1d,but it could significantly enhance the expression of Egr4 after 2d,and reduce the expression of C1 significantly after 2d.The rats administrated with C3aR inhibitor into cerebral ventricle continueously showed the same result in accordance with the treatment of salidroside.And the treatment of salidroside and C3aR inhibitor did not show remarkable additive effects.Conclusion The neuroprotective effect of salidroside on acute cerebral ischemia/reperfusion injury may be related to the inhibition of the activation of the classical pathway of complement,the regulation of Egrs and the reduction of apoptosis.

OBJECTIVETo investigate the effects of losartan on left ventricular hypertrophy (LVH) and plasma transforming growth factor-beta1 (TGF-beta1) in elderly patients with essential hypertension (EH).

METHODSThe elderly patients with EH were divided into two groups, namely EH+LVH group and EH group according to the data of echocardiogram. The systolic and diastolic blood pressures of the patients were monitored. Plasma TGF-beta1 was measured before and after 6 months' treatment with losartan, and the relationship between TGF-beta1 and other index were analyzed.

RESULTSAfter 6 months' treatment, the blood pressure of EH+LVH group and EH group were significantly lowered (P<0.01). Significant improvement of IVSTd, LVPWd, E/A, and LVMI (P<0.01) and obvious reduction of plasma TGF-beta1 (P<0.01) occurred in EH+LVH group after 6 months' treatment. Correlation analyses indicated that the plasma TGF-beta1 level was positively correlated to LVMI (P<0.01).

CONCLUSIONLosartan can reversed LVH in elderly patients with EH partially by lowering plasma TGF-beta1 level.

Aged ; Antihypertensive Agents ; therapeutic use ; Female ; Humans ; Hypertension ; blood ; complications ; drug therapy ; Hypertrophy, Left Ventricular ; blood ; drug therapy ; etiology ; Losartan ; therapeutic use ; Male ; Middle Aged ; Transforming Growth Factor beta1 ; blood

OBJECTIVETo study single nucleotide polymorphism (SNP) of all the coding region in ABCA1 gene in 112 patients with coronary heart diseases.

METHODSWith polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) combining argentation and glue retrieval, DNA sequencing, and restriction fragment length polymorphism (RFLP), the SNP of the 50 exons in all the coding regions of ABCA1 gene was detected in 112 patients with established diagnosis of coronary heart disease.

RESULTSIn the Chinese population with coronary heart disease, besides the SNP variation at R219K and M883I as widely reported, a new single base variation at A1092G in exon 7 was detected, which led to a conversion of the amino-side residue to M223V. This variation was confirmed to represent a novel SNP by RFLP in 108 normal subjects.

CONCLUSIONSThe Chinese population with coronary heart disease has not only the reported SNP changes at the sites R219K and M883I, but also changes at the new SNP site of M233V, which is discovered for the first time in M233V of ABCA1 gene. This variation may increase the risks for coronary heart diseases, and its exact function awaits examination in further epidemiologic survey.

ATP Binding Cassette Transporter 1 ; ATP-Binding Cassette Transporters ; genetics ; Adult ; Aged ; Base Sequence ; Coronary Disease ; genetics ; Female ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Open Reading Frames ; genetics ; Point Mutation ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Polymorphism, Single Nucleotide ; Polymorphism, Single-Stranded Conformational ; Sequence Analysis, DNA