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.

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 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.

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.

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 study the diagnostic value of common inflammatory markers in patients with infectious diseases. Methods One hundred sepsis patients, 100 viral infection patients,100 pulmonary tuberculosis patients and 100 gonorrhea patients were analyzed retrospectively. The contents of procalcitonin (PCT), C-reactive protein (CRP), haptoglobin (HP), ceruloplasmin (CER), α1-acid glycoprotein (α1-AAG), α1-antitrypsin (α1-AAT), white blood cell count (WBC) and erythrocyte sedimetation rate (ESR) were measured. The receiver operating characteristic curve (ROC curve), sensitivity, specificity, positive predictive value, negative predictive value, Youden's index,positive and negative likelihood ratios and total coincidence rate were calculated respectively. Results The area under the ROC curve, sensitivity, specificity, Youden's index and positive likelihood ratios,positive predictive value and total coincidence rate of PCT in sepsis patients were 0. 895, 0.84, 0.92,0.76, 10.50, 0.91 and 0.88, respectively, which were superior to CRP, HP, CER, α1-AAG, α1-AAT, WBC and ESR. Conclusions PCT is a better inflammatory reactive parameter than other parameters currently applied in practice and may serve as a rapid and sensitive test in the early stage of severe bacterial infections.

Objective To investigate the effect of TGFβ1 siRNA on hepatic satellite cells (HSCs) activation, proliferation and extracellular matrix production. Methods The TGFβ1, siRNA plasmid was transfected into HSCs with Lipofectamine 2000. The supernatant and HSCs were collected after incubation for 72h. The expression of TGFβ1, and a-SMA protein in HSCs was detected by. Western blotting. The expression of type Ⅰ and Ⅲ collagen mRNA was detected by RT-PCR. The cell proliferation was assayed by MTT method. Contents of typeⅣ collagen and hyaluronic acid in supernatant were determined by radioimmuno-assay.Results Compared with scrambled control group, the TGFβ1, and a-SMA protein expression,the activity of HSCs proliferation,the expression of typeⅠ and Ⅲ collagen mRNA,and the contents of type Ⅳ collagen and hyaluronic acid in supernatant were reduced in TGFβ1, siRNA group by (79±5)%,(55±4)%, (25±4)% ,(63±6)% ,(57±4)% ,(53±8)% ,(46±8)% ( P<0.01),respectively.Conclusion TGFβ1, siRNA could significantly reduce the expression of TGFβ1,inhibited HSC activation,proliferation and extracellular matrix production.

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

ObjectiveToevaluatethefeasibilityandresultsof transorallaparoscopic thyroidectomy. MethodThyroidectomy was attempted in 5 cases,including 4 females and 1 male with the average age of 42 years( range 35 -60 years).All patients was diagnosed as single nodule of the thyroid gland confirmed by B-mode ultrasound examination before the operation.The average diameter of nodule was 2.5 cm (range 2 - 3.4 cm). ResultTransoral laparoscopic thyroidectomy was perfoormed successfully in all 5 patients.The operation time was 120 - 210 min,averaging 170 min,blood loss during the operation was 15-60 ml,the postoperative hospitalization was 4 days. There was no conversion to open surgery,no recurrent laryngeal nerve injury,nor parathyroid glands dysfunction. ConclusionsTransoral laparoscopic thyroidectomy is feasible and safe,giving excellent cosmetic results.