Adolescent ; Arteriovenous Fistula ; therapy ; Catheter Ablation ; methods ; Coronary Artery Disease ; therapy ; Female ; Heart Defects, Congenital ; therapy ; Humans ; Male ; Middle Aged

Adult ; Female ; Humans ; Myocardial Infarction ; Puerperal Disorders

Adolescent ; Catheterization ; adverse effects ; Endocarditis, Bacterial ; etiology ; Female ; Heart Septal Defects, Ventricular ; therapy ; Humans ; Postoperative Complications

Aged ; Coronary Angiography ; methods ; Coronary Artery Disease ; diagnostic imaging ; Female ; Femoral Artery ; Humans ; Male ; Middle Aged ; Prospective Studies ; Radial Artery

OBJECTIVETo study the changes of morphology and erectile function of the cavernous tissues in spontaneously hypertensive rats.

METHODSSpontaneously hypertensive male rats (SHR) (n = 15) and normotensive Wistar-Kyoto rats (WKY) (n = 15) were studied for 20 weeks. Systolic blood pressure (SBP) was measured weekly by the tail/cuff method. Erectile function was tested by injecting apomorphine (APO). The expression alpha-smooth muscle actin (alpha-SMA) and collagen III was examined by immunohistochemistry.

RESULTSSHR showed a higher systolic blood pressure (205.7 +/- 11.9 vs 114.3 +/- 10.2 mm Hg) and a lower erection frequency (0.6 +/- 0.5 vs 2.4 +/- 0.6). The expression of alpha-smooth muscle actin and collagen III in the cavernous tissues in the SHR was significantly higher than in the WKY.

CONCLUSIONThe erectile function of the penis is markedly affected by hypertension, and the pathological changes may be one of the most important mechanisms of decreased erectile function in SHR.

Actins ; biosynthesis ; Animals ; Apomorphine ; administration & dosage ; Blood Pressure ; physiology ; Collagen Type III ; biosynthesis ; Hypertension ; pathology ; physiopathology ; Immunohistochemistry ; Male ; Penile Erection ; physiology ; Penis ; metabolism ; pathology ; Rats ; Rats, Inbred SHR ; Rats, Inbred WKY

BACKGROUNDRevTet-On gene expression system was used to deliver the suicide gene tk to human breast cancer cell line MCF-7 and control the tk gene expression level. The animal model of human breast cancer on severe combined immune deficiency (SCID) mice was set up to explore the suicide gene therapy by the regulation of Tet-On.

METHODSHerpes simplex virus-thymidine kinase (HSVtk) gene was inserted into the plasmid pRevTRE and the recombinant retroviral vector pRevTRE/HSVtk was constructed. Using modified calcium phosphate co-precipitation method, two transfections, pRevTRE/HSVtk and pRevTet-On were performed for MCF-7 cell line and selected by hygromycin B and G418. MCF-7 cell line that stably expressed Tet-regulated tk gene was established. HSVtk gene expression in the MCF/TRE/tk/Tet-On cell line was under the control of Doxycycline (Dox). Cell viability was also determined by MTT assay, whereas HSVtk gene expression was analyzed by reverse transcription-PCR (RT-PCR).

RESULTSMCF/TRE/tk/Tet-On cell survival rate was decreased from 100% to less than 20% when ganciclovir (GCV) concentration was increased from 0 to 1000 microg/ml at 1 microg/ml of Dox after 72 hours of GCV administration. At 1 microg/ml of GCV concentration, the cell numbers decreased from 7 x 10(4) cells/ml to 2 x 10(4) cells/ml when Dox concentration was increased from 0 to 1500 ng/ml after 72 hours culture. In addition, bystander effects were generated in vitro when 10% - 25% of transduced MCF-7 cells were mixed in untransduced MCF-7 cells. On the other hand, the human breast cancer models in SCID mice were set up. The tk gene was expressed with the regulated character after MCF/TRE/tk/Tet-On cells were implanted into the female SCID mice 7 days after Dox induction followed by intraperitoneally administration of GCV for 23 days. Subcutaneous tumors in SCID mice that were implanted with MCF/TRE/tk/Tet-On cells shrank remarkably after Dox and GCV administration as compared with the control.

CONCLUSIONThe human breast tumor cells (MCF-7) expressing HSVtk gene can be eradicated by administration of GCV and induced with tetracycline or its derivative Dox in vitro and in vivo.

Animals ; Breast Neoplasms ; therapy ; Bystander Effect ; Cell Line, Tumor ; Cell Survival ; Doxycycline ; pharmacology ; Ganciclovir ; pharmacology ; Genes, Transgenic, Suicide ; Genetic Therapy ; methods ; Genetic Vectors ; Herpesviridae ; genetics ; Humans ; Mice ; Mice, SCID ; Retroviridae ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Thymidine Kinase ; genetics ; Transfection

AIM: To explore the expressive role of lipoprotein-associated phospholipase A_2, high sensitive C-reactive protein and matrix metalloproteinase-9 in vulnerable atherosclerotic plaques in a rabbit model. METHODS: Forty eight New Zealand white male rabbits were randomly divided into 4 groups (12 rabbits each): control group, stable plaque group, p53 group, and p53+drug group. Rabbits in control group were fed with a regular diet and underwent sham operation. Rabbits in stable plaque group, p53 group and p53+drug group underwent balloon induced arterial wall injury and then were fed on a diet with 1% cholesterol. The animals were all fed for 3 months, then the rabbits in p53 group and p53+drug group underwent Ad5-CMV p53 transfection at 10th week. Before killed, the animals in p53+drug group underwent pharmacological triggering with Russell's viper venom (RVV) and histamine to induce the rupture of the atherosclerotic plaques. At the 1st day and before sacrifice, the serum was collected for measuring Lp-PLA_2, hs-CRP, MMP-9, HDL, LDL and VLDL. The expressions of Lp-PLA_2, hs-CRP and MMP-9 in tissues were determined by the methods of hybridization and immunohistochemistry. RESULTS: At the end of 12th week, the serum and tissue levels of Lp-PLA_2 and MMP-9 in stable plaque group, p53 group and p53+drug group were significant different from those in control group and in each group at the first day (P<0.05). The serum levels of Lp-PLA_2 and hs-CRP in p53 group and p53+drug group were significantly higher than those in control group and stable group (P<0.05). The serum levels of Lp-PLA_2, hs-CRP and MMP-9 were all significantly different between p53 group and p53+drug group (P<0.05). At the end of 12th week, pathological results showed that 4 groups were normal artery, stable plaque, vulnerable plaque and rupture plaque, respectively. The fabric cap was thicker in plaque groups than that in normal group (P<0.05). The rupture and formation of thrombus were more significant in p53+drug group than those in p53 group. The serum level of Lp-PLA_2 had negative interrelated relationship with fabric cap in plaque groups (r=-0.710, P<0.01), and hs-CRP, MMP-9 had no interrelated relationships with fabric cap in plaque groups. CONCLUSION: Base on the successful establishment of the atherosclerotic plaque animal model, serum Lp-PLA_2 shows better interrelated relationships to plaques stability. Combination with hs-CRP and MMP-9, we can exactly evaluate the nature of plaques.

OBJECTIVETo investigate the effects of hydroxycamptothecin (HCPT) on proliferation and apoptosis of rat hepatic stellate cells (HSC).

METHODSRat HSC line (HSC-T6) and rat hepatocyte line (BRL-3A) were treated with different concentrations of HCPT (0, 0.008, 0.016, 0.031, 0.125, 0.25, 0.5, 1, 2, 4, 8, 16, 32 mg/L respectively) for 24 h. Cell proliferation was assessed by MTT colorimetric assay, apoptosis was detected with PI staging followed by flow cytometry, and by DNA ladder assay. The morphological change of apoptosis was observed under transmission electron microscopy (TEM).

RESULTSMTT assay indicated that HCPT significantly inhibited the proliferation of HSC-T6 and BRL-3A in a dose-dependent manner. 24 h after the treatment with different concentrations of HCPT (0.25, 0.5, 1 mg/L), the apoptosis rate (13.46%+/-2.42%, 26.25%+/-5.65%, 47.05%+/-8.76%, respectively) in HSC-T6 was significantly higher than that in control cells (4.89%+/-1.80%, F = 34.24, P less than 0.01). 24 h after 0.5 mg/L HCPT treatment, cell shrinkage, nucleoli disappearance, chromatin condensation were found under TEM, and DNA ladder was demonstrated by agarose gel electrophoresis.

CONCLUSIONHCPT could significantly inhibit proliferation and induce apoptosis of HSC-T6 in a dose-dependent manner.

Animals ; Apoptosis ; drug effects ; Camptothecin ; analogs & derivatives ; pharmacology ; Cell Line ; Cell Proliferation ; drug effects ; Hepatic Stellate Cells ; drug effects ; Rats

BACKGROUNDPercutaneous vertebroplasty (PVP) has become a popular procedure for painful vertebral osteoporotic fracture (VOF), with immediate pain relief and improved mobility; however, polymethylmethacrylate (PMMA) injected into the vertebral body is not absorbable and little information is available concerning the long-term results. In this retrospective study, we evaluated the long-term clinical results and radiological changes after PVPs for VOFs.

METHODSFifty-one patients with VOFs were treated by PVPs with PMMA between 2000 and 2004. After > 7 years of follow-up, eight patients had died from causes unrelated to the intervention and 12 patients were lost to follow-up, thus leaving 31 patients available for evaluation with an average length of follow-up of 9.2 years (follow-up rate, 72.1%). Among these 31 patients, the PMMA was injected at 43 levels with a mean volume of 4.3 ml per level (range, 2 - 6 ml). The pain was assessed with a visual analog scale (VAS), and the mobility was graded as walking without difficulty (grade 1), walking with assistance (grade 2), and bedridden (grade 3). Plain radiographs and computed tomography (CT) were obtained and assessed pre-operatively, immediately post-operatively, and after 7 years of follow-up. The PMMA, vertebral height, and Cobb angle were assessed and compared.

RESULTSAll of the patients experienced pain relief and improved mobility after intervention and during the follow-up period. Cement leakage was detected in post-operative CT scans in 9 of 51 patients, but without neurological compromise. For the 31 patients followed up over 7 years, the VAS decreased from 8.3 ± 2.6 pre-operatively, to 2.1 ± 1.6 immediately post-operatively, and 1.0 ± 0.9 at the final follow-up evaluation, with significantly improved mobility. Additional compression fractures occurred at adjacent levels in three patients, and there were no new fractures at the augmented vertebrae. Based on a review of the radiographs, neither loose nor displaced cement was detected. The changes in vertebral height and Cobb angle were not significant. On CT scans, the cement closely contacted or infiltrated the trabecular bone. The boundary between the cement and trabecular bone was indistinct and there was no evident radiolucent gap between the cement and trabecular bone.

CONCLUSIONSAt an average follow-up of 9.2 years, PVPs provided sustained pain relief and improved mobility in patients with VOFs. The PMMA injected into the vertebral body combined closely with the host trabecular bone without adverse reactions.

Aged ; Aged, 80 and over ; Female ; Humans ; Male ; Middle Aged ; Osteoporotic Fractures ; surgery ; Polymethyl Methacrylate ; therapeutic use ; Retrospective Studies ; Spinal Fractures ; surgery ; Vertebroplasty ; methods

OBJECTIVETo explore the HSVtk gene expression mediated by the retroviral vector and to obtain high titer recombinant retroviral virus.

METHODSThe recombinant vector pRevTRE/HSVtk was constructed by inserting HSVtk gene into pRevTRE. The recombinant retrovirus, which was produced from cloned PA317 cells screened by hygromycin B after "micro-pingpong" technique transferring with pRevTRE/HSVtk plasmids DNA by using modified calcium phosphate precipitation method. HSVtk gene expression was performed on target cells and virus titers were detected in different cultured temper, time and sodium butyrate concentration.

RESULTSThe recombinant retroviral vector pRevTRE/HSVtk was constructed and HSVtk gene expression was detected on target cells after they were infected with the recombinant retrovirus.

CONCLUSIONHigh titer of retroviruses could be obtained in the culture medium of PA317 cell line through "micro-pingpong" technique at 30 hours and 10 mmol/L sodium butyrate concentration followed by frozen ultrafiltration.

Animals ; Breast Neoplasms ; enzymology ; pathology ; virology ; Cell Line ; Cell Line, Tumor ; Female ; Gene Expression Regulation, Enzymologic ; Genetic Vectors ; Humans ; Mice ; NIH 3T3 Cells ; Recombination, Genetic ; Retroviridae ; genetics ; Simplexvirus ; enzymology ; genetics ; Thymidine Kinase ; biosynthesis ; genetics ; Titrimetry ; Transfection