Objective To evaluate the possibility and efficiency of nanoparticles as a new vector in vascular endothelial growth factor (VEGF) gene transfection. Methods Nanoparticle-VEGF (Np/VEGF)complex was prepared with poly (D, L-lactide-co-glycolide) (PLGA) loading VEGF165 gene using the multiple emulsion (w/o/w) technique. The envelopment efficiency and size of the complex were determined. Rat myocardial cells were cultured in vitro, and the Np/VEGF was transfected into the cultured myocardial cells. Then RT-PCR and ELISA were used to evaluate whether the Np/VEGF increased the level of gene expression. Four New Zealand rabbits were used, the suspension of Np/VEGF was injected into myocardial tissue of rabbits after thoracotomy. 96h after the operation, the tissue sections of the implant sites were observed with transmission electron microscope (TEM) to determine the process of nanoparticles as vectors for gene transfer to cardiac myocytes. Results The envelopment efficiency and size of the Np/VEGF complex thus prepared were 1.87% and 25-300nm respectively. RT-PCR and ELISA showed that VEGF gene could be successfully transfected into myocardial cells by nanoparticle, and NP/VEGF significantly enhanced gene transfection efficiency, and it was more effective than plasmid. 96h after the operation, a great number of nanoparticles were observed in myocardial cytoplasm and nucleus with TEM, and many nanoparticles began to dissolve and degrade, suggesting that the DNA was released slowly from the nanoparticles localized in the cytoplasmic compartment, and was then transferred into the nucleus. Conclusions NP/VEGF can act as a vector to transfect VEGF gene in vitro and in vivo, it significantly enhanced gene transfection efficiency, and it was more effective than plasmid.

Objective To investigate the effect of microRNA-21 (miR-21) antisense oligonucleoti-de on collagen synthesis in the rat hepatic satellite cells ( HSCs) .Methods Rat hepatic stellate cells were isolated and cultured; the miR-21 antisense oligonucleotide was transfected into HSCs with lipofectamine 2000;after incubation 48 h, the HSCs were collected.The expression of miR-21 was detected with reverse transcription polymerase chain reaction ( RT-PCR) , andα-smooth muscle actin (α-SMA) protein in HSCs with Western blot.The cell proliferation was assayed with methyl thiazolyl tetrazolium ( MTT) method.Re-sults Compared to scrambled control group, the expression of miR-21 was reduced by 76%( P <0.01), the proliferation activity of HSCs was reduced by(26 ±3)%( P <0.01), the expressions of type I and III collagen proteins were reduced by(61 ±7)%and (48 ±6)%( P <0.01).Conclusions The miR-21 an-tisense oligonucleotide could reduce significantly the expression of miR-21, and inhibit HSC proliferation and extracellular matrix production.

Antineoplastic Agents ; administration & dosage ; adverse effects ; Asparaginase ; administration & dosage ; adverse effects ; Child ; Child, Preschool ; Combined Modality Therapy ; Dose-Response Relationship, Drug ; Drug Hypersensitivity ; prevention & control ; Female ; Humans ; Hyperglycemia ; chemically induced ; prevention & control ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; drug therapy ; Severity of Illness Index ; Time Factors ; Treatment Outcome

Objective To investigate the effect of miRNA-200b-specific inhibitor on hepatic stellate cells(HSCs) activation,proliferation,and extracellular matrix production.Methods The miRNA-200b-specific inhibitors were designed,synthesized,and transfected into HSCs with lipofectamine 2000.The supernatant and HSCs were collected after incubation for 48 h.The expression of miR-200b was detected by quantitative reverse transcription polymerase chain reaction (qRT-PCR).The expression ofα-smooth muscle actin (oα-SMA) protein in HSCs was detected by Western blotting.The cell proliferation was assayed by methyl thiazolyl tetrazolium (MTT) method.Contents of type Ⅲ procollagen and hyaluronic acid in supernatant were determined by radioimmunoassay.Results Compared to the control group,miRNA-200b expression was decreased in the miRNA-200b inhibitor group by 82％ (P ＜ 0.01),α-SMA protein expression was reduced in the miRNA-200b inhibitor group by (19 ± 3) ％ (P ＜ 0.05),and the activity of HSCs proliferation was reduced by(33 ± 5)％ (P ＜0.01),and the contents of type Ⅲ procollagen and hyaluronic acid in supernatant were reduced in miRNA-200b inhibitor group by (35 ± 4)％ and (31 ± 2)％,respectively(P ＜0.01).Conclusions The miRNA-200b-specific inhibitor could significantly reduce the expression of miRNA-200b,and inhibit HSC proliferation,activation,and extracellular matrix production.

OBJECTIVETo observe the clinical efficacy of Wangbi Tablet (WT) on the treatment of knee osteoarthritis (KOA), and to study its treatment program by integrative medicine.

METHODSThe randomized, parallel control, and multi-center clinical observation method was used. 160 KOA patients of Gan-Shen deficiency complicated with stasis-blood blockade syndrome were assigned to three groups. Group A (72 cases) was treated by WT, Group B (42 cases) was treated by Votalin Tablet, and Group C (46 cases) was treated by WT + Votalin Tablet. All patients were treated for two months. Before and after treatment the clinical symptoms, the knee function activity, adverse reactions were observed and recorded. Erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) of each patient were respectively detected, statistically analyzed and compared.

RESULTSAfter eight weeks of treatment significant difference was shown in the time of morning stiffness in joints, joint tendemess, swelling, pain, joint functional activities, ESR, and CRP between Group A and Group B (P<0.01). Besides, better effects were obtained in Group C (P<0.01). Improvement of ESR and CRP: They were obviously improved in the three groups after treatment (P<0.01). As for the markedly effective rate, better effects were obtained in Group C (P<0.05).

CONCLUSIONSIn the treatment of KOA, WT showed favorable effects with no obvious adverse reaction. The combination of WT and Voltaren Tablet was significantly superior to use of WT or Votalin Tablet alone. It could reduce the dosage of Voltaren Tablet and avoid adverse reactions of the gastrointestinal tract.

Adult ; Aged ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Integrative Medicine ; Male ; Middle Aged ; Osteoarthritis, Knee ; drug therapy ; Phytotherapy ; Tablets

OBJECTIVEExtracellular signal-regulated kinases (ERKs) can be activated by calcium signals. In this study, we investigated whether calcium-dependent kinases were involved in ERKs cascade activation after global cerebral ischemia.

METHODSCerebral ischemia was induced by four-vessel occlusion, and the calcium-dependent proteins were detected by immunoblot.

RESULTSLethal-simulated ischemia significantly resulted in ERKs activation in N-methyl-D-aspartate (NMDA) receptor-dependent manner, accompanying with differential upregulation of Src kinase and Ca2+/calmodulin-dependent protein kinase II (CaMKII) activities. With the inhibition of Src family tyrosine kinases or CaMKII by administration of PP2 or KN62, the phosphorylation of ERKs was impaired dramatically during post-ischemia recovery. However, ischemic challenge also repressed ERKs activity when Src kinase was excessively activated.

CONCLUSIONSrc family tyrosine kinases and CaMKII might be involved in the activation of ERKs mediated by NMDA receptor in response to acute ischemic stimuli in vivo, but the intense activation of Src kinase resulted from ischemia may play a reverse role in the ERKs cascade.

Analysis of Variance ; Animals ; Brain Ischemia ; enzymology ; pathology ; Calcium-Calmodulin-Dependent Protein Kinase Type 2 ; Calcium-Calmodulin-Dependent Protein Kinases ; metabolism ; Disease Models, Animal ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Gene Expression Regulation ; physiology ; Hippocampus ; cytology ; enzymology ; Male ; Neurons ; enzymology ; pathology ; Phosphorylation ; Rats ; Rats, Sprague-Dawley ; Receptors, N-Methyl-D-Aspartate ; metabolism ; Signal Transduction ; physiology ; Statistics, Nonparametric ; src-Family Kinases ; metabolism

Antineoplastic Agents ; therapeutic use ; Apoptosis ; drug effects ; Caspase Inhibitors ; Drug Resistance, Neoplasm ; Gene Expression Regulation, Neoplastic ; Inhibitor of Apoptosis Proteins ; metabolism ; Neoplasms ; drug therapy ; metabolism ; pathology

Objective To evaluate the outcome of surgical treatment of tricuspid valve regurgitation after valve replacement.Methods Twenty one patients with tricuspid valve insufficiency after valve replacement were performed surgical treatment.Tricuspid valve-plasty was performed in 17 patients and tricuspid valve replacement was done in 4 patients.Three patients underwent edge-to-edge tricuspid valveplasty.DeVega procedure was performed in 5 patients.Cosgrove-Edward annuloplasty ring was used in 15 patients.Four patients' valve were replaced by SJM bileallet mechanical prostheses.Results There was 3 patients dead early after operation with a mortality of 14.3％(3/21).The causes of death including multiple organ failure,cardiac arrest and low cardiac output syndrome.The rate of early postoperative complications was 33.3％ (7/21),including pulmonary edema,arrhythmia,acute renal failure and low cardiac output syndrome.The patients were followed up 9-60 months.The rate of readmission was 23.8％ (5/21).Two patients went to hospital again for pleural effusion,1 patient for left ventricular dysfunction and 2 patients for right ventricular dysfunction.The other patients recovered well.Conclusions Surgical therapy is effective on severe tricuspid valve regurgitation after valve replacement.Preoperative aggressive treatment of heart failure,a reasonable grasp of surgical indications and timing of surgery,strict perioperative management are the keys to guarantee patients a smooth recovery.

Objective To analyse the difference of effects of different tidal volume mechanical ventilation strategies in infants with severe congenital heart disease after curative.Methods Sixty-eight cases were chosen from CICU of Anhui Provincial Children's Hospital during Oct 2010 to Jan 2012.Thirty-two cases in A group were used in ventilation strategy by lower tidal volumes (6 ～ 10 ml/kg),36 cases in B group were used in ventilation strategy by larger tidal volumes (10 ～ 15 ml/kg).The time of mechanical ventilation,duration of ICU stay and postoperative complications were compared between the two groups.Results The time of mechanical ventilation of B group was shorter than that of A group [(8.6 ± 2.5) d vs (11.7 ± 3.2) d],duration of ICU stay of B group was shorter than that of A group [(11.4 ± 4.8) d vs (15.6 ± 5.7) d],there were statistical differences between two groups(P ＜ 0.0l).The incidence rate of ventilator associated pneumonia and pneumothorax were 6.3％ and 3.1％ in A group,which were 5.6％ and 5.6％ in B group,there were no statistical differences between two groups (P ＞ 0.05).Conclusion The effects of mechanical ventilation with larger tidal volumes is better than that with lower tidal volumes in infants with congenital heart disease postoperative therapy.