BACKGROUND:Currently, there are few reports on the effects of Caspase inhibitors combined with Cocktail protease inhibitors on isolation and purification of islet cel s. OBJECTIVE:To explore the effects of Caspase inhibitors and Cocktail protease as protectors on islet cel s during isolation and purification process. METHODS:New-born pigs were selected to separate, purify and culture the islet cel s, and after 24 hours of culture, the specimens were divided into three groups:blank control group, experimental group A (Caspase inhibitors and Cocktail protease were added only during digestion process), and experimental group B (Caspase inhibitors and Cocktail protease were added both during digestion and culture processes). AO-EB staining was used to qualitatively observe cel morphology and apoptosis. Flow cytometry was employed to quantitatively detect cel viability and apoptosis. RESULTS AND CONCLUSION:Percentages ofβcel s were 66.91%in blank control group, 84.58%in the experimental group A, and 87.15%in the experimental group B. The proportions of living cel s, apoptotic cel s and dead cel s were 56.52%, 16.15%, 21.25%in the blank control group, 62.27%, 14.66%, 14.47%in the experimental group A, and 73.09%, 6.83%, 10.28%in the experimental group B, respectively. These findings indicate that in the process of digestion and culture in vitro, Caspase inhibitors and Cocktail protease inhibitors can both obviously decrease the loss of cel s, and increase the percentage of islet beta cel s.

Objective To discuss the treatment efficacy and radiotherapy side effects of the preoperative long-course radiochemotherapy and preoperative short-course radiotherapy.Methods 64 patients with local advanced middle and low rectal cancer who got the treatment from April 2004 to April 2010 were analyzed retrospectively.40 patients got the preoperative long-course radiochemotherapy under the dose of DT 45-50 Gy/25 F,1.8-2.0 Gy/F,5 F/W,combining with the synchronous capecitabine chemotherapy (1 650 mg/m2,2 F/d,d1-14/d21-35),and accepted operation 4-6 weeks after the radiotherapy.The rest 24 patients underwent the preoperative short-course radiotherapy under the dose of DT 25 Gy/5 F,5 Gy/F,5 F/W,and got the operation in 2 weeks after the radiotherapy.Results The radical and anus reservation rates in preoperative long-course radiochemotherapy group [85.0 ％ (34/40),65.0 ％ (26/40)] were higher than those in preoperative short-course radiotherapy group [58.3 ％ (14/24),33.3 ％ (8/24)] (x2 =5.689,P =0.019;x2 =6.040,P =0.041).There were no significant differences between the two groups on the index of remission rates,radiation injury,surgical complications,and overall survival rate of 1,3,5 years (all P ＞ 0.05).Conclusions The remission rate and overall survival time between the preoperative long-course radiochemotherapy group and preoperative short-course radiotherapy have no significant difference.But the preoperative long-course radiochemotherapy may improve the anus reservation rate and the radical resection rate,without increasing the radiation injury and surgical complications.

OBJECTIVETo investigate the effects of growth hormone (GH) on the erectile function and the number of neuronal nitric oxide synthase (nNOS) -containing nerve fibers in the penis of aged rats.

METHODSTwenty-four aged male SD rats (18 months) were randomized into 2 groups: GH intervention group and control group. After four and eight weeks, a half of each group were selected and tested for erectile function after apomorphine (APO) injection and then sacrificed for the detection of nNOS-containing nerve fibers in the penis by streptavidin-peroxidase conjugated method (SP method).

RESULTSAfter four weeks, the erectile function and the number of nNOS-containing nerve fibers showed no significant difference between the GH intervention group and the control group (P > 0.05). After eight weeks, the erection frequency was significantly different (P < 0.05) between the two groups, while the erection rate was not. The number of nNOS-containing nerve fibers in the GH intervention group was significantly larger than that in the control group (P < 0.01).

CONCLUSIONGH enhances the regeneration of nNOS-containing nerve fibers in the penis and improves the erectile function of the aged rat.

Animals ; Apomorphine ; pharmacology ; Human Growth Hormone ; pharmacology ; Immunohistochemistry ; Male ; Nerve Fibers ; enzymology ; physiology ; Nerve Regeneration ; drug effects ; Nitric Oxide Synthase Type I ; analysis ; Penile Erection ; drug effects ; Penis ; enzymology ; innervation ; Random Allocation ; Rats ; Rats, Sprague-Dawley

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

OBJECTIVETo investigate the changes in peripheral blood bone marrow stem cells and tumor necrosis factor-alpha gene expression in the ischemic myocardium in rabbit models of hibernating myocardium.

METHODSTwenty-four male Japanese white rabbits were randomized into 4 groups, including a sham-operated group and 3 model groups with hibernating myocardium induced by partial ligation of the left anterior descending coronary artery. The percentage of CD34-positive cells in the peripheral blood was evaluated by flow cytometry, and TNF-alpha mRNA expression in the ischemic myocardium was determined by real-time RT-PCR in the 3 model groups (at 3, 7, or 28 days after the operation) and in the sham-operated group.

RESULTSIn rabbits with partial ligation of the left anterior descending coronary artery, the percentage of CD34-positive cells in the peripheral blood and myocardial TNF-alpha mRNA expression were significantly increased at 3 and 7 days after the operation in comparison with those in the sham-operated group and those at 28 days postoperatively (P<0.01). No significant differences were found in the percentage of CD34 positive cells or myocardial TNF-alpha mRNA expression between the sham-operated group and the rabbits 28 days after the coronary artery ligation (P>0.05).

CONCLUSIONBone marrow stem cell can be mobilized into the peripheral blood in rabbit hibernating myocardium model possibly by increasing TNF-alpha gene expression in the ischemic myocardium.

Animals ; Antigens, CD34 ; metabolism ; Bone Marrow Cells ; cytology ; Coronary Vessels ; surgery ; Disease Models, Animal ; Gene Expression Regulation ; Hematopoietic Stem Cell Mobilization ; Hibernation ; Ligation ; Male ; Myocardial Ischemia ; metabolism ; physiopathology ; surgery ; therapy ; RNA, Messenger ; genetics ; metabolism ; Rabbits ; Tumor Necrosis Factor-alpha ; genetics

Adenovirus vectors are one of the most promising gene transfer systems. They are of great value for gene therapy because these vectors achieve temporal high-level transgene expression and high gene transfer efficiency. To meet increasing needs of adenovirus vectors for gene therapy programs, parallel development of efficient, scalable and reproducible production processes is required. Perfusion cultivation of 293 cells is one of the most commonly used methods to produce adenovirus vectors and it is suitable for industrialized production specially. Experimental studies had been carried out to produce recombinant adenovirus containing the green fluorescent protein gene (Ad-GFP) by perfusion cultivation of HEK-293 N3S cells in a 5L stirring bioreactors. Perfusion rate was 1-2 volume/day. To infect the 293 N3S cells with Ad-GFP at the density of (2-4) x 10(6) cells/ ml. The time of collecting cells was 48 hours post infection. After three rounds of freeze/thaw and centrifugation, the crude viral lysates were stored at--80 degrees C until use. Then to get the Ad-GFP products by 2 x CsCl-gradient purification. The purity of the products was determined by the A260/A280 ratio and a high performance liquid chromatography (HPLC) assay. The infective titer was determined by a TCID50 assay. The culture term was 10-12 days. The infectious titer, the number of virus particle and the ratio of infectious titer to virus particle for the product were 1.0 x 10(11) IU/mL, 1.68 x 10(12) VP/mL and 6.0% IU/VP respectively. The A260/A280 ratio was 1.33, and the purity determined by HPLC was 99.2%. The cell specific productivity was around 1000 IU/cell. By perfusion cultivation of 293 N3S cells in a 5L stirring bioreactors, we established the production process for Ad-GFP, which paves a way to produce other recombinant adenovirus for gene therapy.
Adenoviridae ; genetics ; growth & development ; isolation & purification ; Bioreactors ; microbiology ; Cell Line ; Gene Transfer Techniques ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; Humans ; Kidney ; cytology ; virology ; Recombinant Proteins ; biosynthesis ; genetics ; Recombination, Genetic ; Virus Cultivation ; instrumentation ; methods

OBJECTIVE: To assess the effects of intracoronary diltiazem on no-reflow phenomenon of infarct-related artery (IRA) after emergent percutaneous transluminal coronary angioplasty or/and intracoronary stenting (PTCA/Stenting) in the patients with acute myocardial infarction (AMI). METHODS: We studied 34 AMI patients with no-reflow phenomenon of IRA after emergent PTCA/Stenting between January 1999 and August 2005. Urokinase-treated group (n=16) was given intracoronary urokinase 30,0000 - 50,0000 units within 15 - 30 minutes between January 1999 and April 2002 while diltiazem-treated group (n=18) was given intracoronary diltiazem 0.5 - 2 mg within 10 - 30 minutes between May 2002 and August 2005. Fifteen minutes later, coronary arteriography (CAG) was performed and the thrombolysis in myocardial infarction (TIMI) flow grade was measured. RESULTS: No apparent change of TIMI flow grade was found between pre-administration and post-administration of intracoronary urokinase, but TIMI flow grade was significantly improved after intracoronary diltiazem (P<0.01). TIMI flow grade of diltiazem-treated group was significantly higher than that of urokinase-treated group after the administration (P<0.05). The percentage of the patients who reached TIMI flow grade 3 after the intracoronary administration was higher in the diltiazem-treated group than that in the urokinase-treated group (P<0.01). CONCLUSION The intracoronary administration of diltiazem 0.5~2mg can effectively improve the no-reflow phenomenon after emergent PTCA/Stenting in patients with AMI.
Adult ; Aged ; Angioplasty, Balloon, Coronary ; Diltiazem ; administration & dosage ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Myocardial Infarction ; therapy ; No-Reflow Phenomenon ; drug therapy ; Stents ; Treatment Outcome

OBJECTIVE: To determine the effect of different doses of atorvastatin on the serum soluble intercellular adhesion molecules-1 (sICAM-1) in patients undergoing percutaneous coronary intervention (PCI). METHODS: The study consisted of 38 patients with unstable angina and 10 patients with old infarction who underwent elected PCI for stenotic lesions of the coronary artery. Patients were randomly assigned to either aggressive group or conventional one. After PCI the patients took atorvastatin 20 mg per day or 10 mg per day. Blood lipid profile was examined before, and 3 months after the PCI. SICAM-1 was examined before the PCI, 48 hours and 3 months after the PCI. RESULTS: The total cholesterol and LDL-Cholesterol 3 months after the PCI in the 2 groups were lower than those before the PCI (P<0.01). The aggressive group showed greater reduction in concentrations of TC and LDL-C than the conventional group (P<0.01). The changes in concentrations of HDL-C between pre-PCI and 3 months after the PCI and TG were not obvious (P>0.05). sICAM-1 in the 2 groups 48 hours after the PCI significantly higher than that before the PCI (P<0.01). But sICAM-1 in the 2 groups 3 months after the PCI significantly lower than that before the PCI (P<0.01 or P<0.05). The aggressive group showed greater reduction than the conventional group (P<0.01). TC and LDL-C were positively correlated with sICAM-1(r=0.2413, r=0.2691, all P<0.05). CONCLUSION Atorvastatin 20 mg per day reduces TC, LDL-C, and sICAM-1 to a greater extent than atorvastatin 10 mg per day. The effect on sICAM-1 is partly related to reduce lipid profile.
Aged ; Atorvastatin ; Female ; Heptanoic Acids ; administration & dosage ; therapeutic use ; Humans ; Intercellular Adhesion Molecule-1 ; blood ; Male ; Middle Aged ; Percutaneous Coronary Intervention ; Pyrroles ; administration & dosage ; therapeutic use

OBJECTIVE: To access the possibility, methods and efficacy of simultaneous transcatheter therapy for ventricular septal defect ( VSD ) combined with atrial septal defect (ASD). METHODS: In 68 patients with VSD, four patients ranging from 3 to 24 years old were combined with ASD. The diameters of perimembranous VSD were 2 approximately 10.5 mm, and the diameters of secundum ASD were 4.6 approximately 7 mm under the echocardiography before the operation. Another 4 patients with VSD occluded by left ventriculography: 3 patients were occluded by VSD occluder first, and then occluded by ASD occuder. The other was only occluded by VSD occluder. RESULTS: All VSD was treated successfully at one time in 4 patients. The diameters of VSD occluder were 4, 8, 10, and 16 mm. ASD was occluded successfully at one time in 3 patients. The diameters of ASD occluder were 8, 10, and 10 mm. The successful rate of the operation was 100%. No complication occurred in the operation and follow-up. CONCLUSION Simultaneous transcatheter closure for VSD combined with ASD is a safe, feasible and effective therapy.
Abnormalities, Multiple ; therapy ; Adolescent ; Adult ; Balloon Occlusion ; instrumentation ; methods ; Cardiac Catheterization ; methods ; Child ; Child, Preschool ; Echocardiography ; Female ; Heart Septal Defects, Atrial ; therapy ; Heart Septal Defects, Ventricular ; therapy ; Humans ; Male ; Treatment Outcome

OBJECTIVE: To evaluate the short and mid-term changes of the cardiac morphology after percutaneous transcatheter closure of ventricular septal defects (VSD) with transthoracic echocardiography (TTE). METHODS: The left ventricular end-diastolic diameter (LVEDD), left ventricular end-diastolic volume (LVEDV), left atrial diameter (LAd), and right ventricular diameter (RVd) in 30 VSD patients were measured before the VSD closure,and on the 3rd day, 3rd month, and 6th month after the VSD closure by TTE. RESULTS: LVEDD and LVEDV significantly decreased on the 3rd day after the VSD closure compared with pre-VSD closure. LVEDD and LVEDV continuously decreased on the 3rd month and 6th month after the VSD closure. LAd was smaller on the 3rd month and 6th month after the VSD closure, but there was not significant difference between the 3rd and 6th month. RVd increased on the 3rd day after the VSD closure, while no significant difference was found among the 3rd month and 6th month before and after VSD closure. CONCLUSION Percutaneous transcatheter VSD closure may effectively improve the cardiac remodeling in VSD patients in the short and mid-term follow-up.
Adolescent ; Adult ; Cardiac Catheterization ; methods ; Child ; Child, Preschool ; Echocardiography ; Female ; Follow-Up Studies ; Heart Septal Defects, Ventricular ; diagnostic imaging ; therapy ; Humans ; Male ; Prosthesis Implantation ; methods ; Time Factors ; Ventricular Remodeling