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 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 ; 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 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.

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.

0bjective To establish a standard curve method with more accuracy employed in fluorescence real-time PCR(RT-PCR)as a alternation of the general method.Methods β-actin and KLK11 plasmid DNA for quantitative standard curve were constructed in our study,and Plasmids of β-actin was employed as a internal control.After serial dilution these plasmid were used as DNA standard to obtained slope.Expression of these two genes in malignant prostate cancer cell line LNCAP were tested by real-time PCR,and we analyzed the RT-PCR results with two different methods and compared their accuracy.Results Thestandards curves made from these linear DNA standards showed good linearity (R2=0.991 and 0.992 for β-actin and KLK11 standards graphs),but also displayed a discrepancy in their PCR efficiency(β-actin 123% and KLK11 99%).There were different results after two different stand curve analytical method:the expression of KLK11 mRNA in LNCAP was downregulated in general standard curve method.In the new analytical method,howerer,KLK11 upregnlated for 4.46-fold.And there was a significant difference between aplification efficiency of targt gene and internal control gene(t=4.829,P<0.05).Conclusions Compared with general standard curve method,the new advanced standard curve method described here avoids an error which considers there is identical amplification efficiency between target gene and internal reference gene.It is considered to be a more correct analytical method in fluorescence real-time PCR.

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.

Objective To construct a large animal model of ischemic heart disease induced by acute myocardial infarction (AMI) . Methods Male Diannan Mini-pigs were selected (n=8), weight 20 ± 4 Kg. After general anesthesia and endotracheal intubation, blood pressure and electrocardiograph were monitored. Parasternal incision was taken and thoracotomyv was performed at 4th-to-5th intercostalgap. Pericardium was opened and left anterior descending artery was ligated. Echocardiography was performed at baseline, postoperative 28 and 60 day. Euthanasia and heart explanted was conducted at 60 day of experimental pig. After gross examination of the heart, the histological examination was used to assess myocardial infarction by using H&E staining. Results All of the pigs were presented with ventricular arrhythmias in varying degrees after ligation of the left anterior descending artery, of whom 2 died of refractory ventricular fibrillation, the mortality was 25%. Echocardiography assessment showed the loss left ventricular function, the thinning of left ventricular apical and the enlargement of left ventricule. Histological examination revealed that myocardial fibrosis and fibrous scar were formed in myocardial infarction area. Conclusion The ligation of left anterior descending artery through thoracotomy under direct vision is a reliable strategy for construction of AMI porcine model and the experimental porcine can develop ischemic cardiac dysfunction progressively.