BACKGROUND: At present, bladder acellular matrix grafts have been successfully used for substituting animal bladder and urinary canal, and for repairing hypospadia. However, reports on bladder acellular matrix grafts for substituting albuginea penis need to be investigated. OBJECTIVE: Allogeneic bladder acellular grafts were used for substituting albuginea penis of rabbits, in order to observe repairing results. DESIGN: A randomized controlled observation. SETTING: West China Medical Laboratory Animal Center and West China Laboratory of Tissue Engineering of Sichuan University as well as Laboratory of Tissue Engineering of Guiyang Medical College. MATERIALS: Fifty male healthy New Zealand Rabbits of grade 3, weighing 2.6-3.0 kg, without phimosis and penis dysplasia, and without presence of phallocampsis after normal saline being perfused, were provided by Huaxi Laboratory Animal Center of Sichuan University. METHODS: This study was performed at the West China Laboratory Animal Center and West China Laboratory of Tissue Engineering of Sichuan University as well as Laboratory of Tissue Engineering of Guiyang Medical College between December 2005 and June 2007. Bladders were taken from 10 experimental rabbits for preparing bladder acellular matrix grafts. The other 40 New Zealand rabbits were randomly divided into the control group, and the bladder acellular matrix grafts group, with 20 in each. An area of 10 mm×5 mm of albuginea penis was resected from dorsum penis of each rabbit. Suture in situ of albuginea penis and bladder acellular matrix grafting were conducted in rabbits of the control group and bladder acellular matrix grafts group, respectively. In the 2nd, 6th, 12th and 24th weeks postoperatively, each rabbit was intracavernously perfused normal saline for inducing penile erection, separately, in order to observe phallocampsis. At above-mentioned each time point, experimental animals were sacrificed. Sample was taken from surgical region for haematoxylin-eosin (HE) staining and Masson trichrome staining, in order to observe the changes of tissue and structure of surgical region. Types Ⅰand Ⅲ collagen fiber areas were detected by Stirus red staining, and the expressions of inducible nitric oxide synthase(iNOS) and transforming growth factor-β1 (TGF-β1) were detected by immunohistochemical staining. MAIN OUTCOME MEASURES: ①Phallocampsis status. ② Changes of tissue and structure of surgical region. ③iNOS and TGF-β1 expressions. ④TypeⅠand Ⅲ collagen fiber areas.RESULTS: Forty experimental rabbits were involved in the penile surgery, two of them died from overdose anesthesia, two died from chordapsus, so the remaining thirty-six rabbits were involved in the final analysis. In the 6th week postoperatively, phallocampsis reached its highest level, and 2 rabbits in the control group and 1 rabbit in the bladder acellular matrix grafts group presented phallocampsis. In the 12th week, every rabbit presented phallocampsis. In the 24th week, 1 rabbit in the control group but none in the bladder acellular matrix grafts group presented phallocampsis. In the 2nd week, the structure of surgical regions of each rabbit was poorly clear, with remarkable inflammatory infiltration. In the bladder acellular matrix grafts group, grafting regions presented cells ingrowing the bladder acellular matrix grafts. Masson trichrome staining results showed that in the surgical region, tunica albuginea fibers were thin and poorly arranged. In the 6th week, tunica albuginea recovered its integrity, and bladder acellular matrix grafts could not be distinguished. No significant difference existed between two groups. In the 24th week, tunica albuginea was even and complete in the sugical region, and fibers restored their arrangement of circular muscle in inner layer and longitudinal muscle in outer layer, without difference from normal tunica albuginea. iNOS and TGF-β1 expressions were the strongest in the 2nd week, and they were found in the fibrocytes and vascular endothelial cells in the 6th week, but a little in the 12th and 24th weeks postoperatively. There were no remarkable differences in iNOS and TGF-β1 expressions between two groups at the same time point. In the 2nd week, typesⅠand Ⅲ collagen fibers co-existed with equivalent proportion. Then, typeⅠcollagen fibers were gradually increased, while type Ⅲ collagen fibers were on the contrary. In the 24th week, typeⅠcollagen fibers took the main place and type Ⅲ collagen fibers were unremarkable. CONCLUSION: Bladder acellular matrix grafts have no remarkable inflammatory reactions and fibrosis in repairing tunica albuginea of New Zealand rabbits, so they are very ideal grafting materials for penile surgery.

AIMTo approach the effects of multi-site synchronous ventricular pacing on myocardial mechanics and cardiac work.

METHODSFive modes of multi-site synchronous ventricular pacing were randomly performed in 12 dogs with anesthetized, opened chest and artificial-ventilation. Some parameters were measured simultaneously including: the peak of left ventricular pressure rise and fall (+/- dp/ dt(max)), the time constant of left ventricular relaxation(tau), the muscle tensile force in left/right ventricular wall (V-tensile force, V-TF), SV, LVSW and RVSW.

RESULTSThe myocardial systolic mechanical parameters: +dp/dt(max) and LV-TF of cHisB-LVPL and RVA-LVPL pacing by biventricular pacing modes were increased than that of cHisB-RVA pacing in right ventricular bifocal pacing mode. +dp/dt(max) in above two groups of biventricular pacing was increased than that in cHisB-RVA pacing. Tau value of cHisB-LVPL and RVA-LVPL pacing modes were shorted than that of cHisB-RVA pacing. The above parameters of cHisB-RVA-LVPL and cHisB-RVA-LVA biventricular trifocal pacing were superior to that of cHisB-LVPL and RVA-LVPL biventricular pacing. The +dp/dt(max), LV-TF and RV-TF of cHisB-RVA-LVPL pacing were increased as compared with that of cHisB-RVA-LVA pacing (P > 0.05). The -dp/dt(max) in cHisB-RVA-LVPL pacing were increased by 6.0% and tau value was shorted by 3.7% compared with those in cHisB-RVA-LVA pacing (P > 0.05). SV, LVSW and RVSW of cHisB-LVPL and RVA-LVPL biventricular pacing were increased than those of cHisB-RVA bifocal pacing. The above parameters of cHisB-RVA-LVPL pacing were increased than that of cHisB-RVA-LVA and cHisB-LVPL pacing.

CONCLUSIONIt was explained that the cHisB-RVA-LVPL biventricular trifocal sites synchronous pacing mode would increase the velocity of ejection and filling during myocardial contraction and relaxation and enhance cardiac work by maintaining normal VSS.

Animals ; Cardiac Pacing, Artificial ; methods ; Dogs ; Female ; Heart ; physiology ; Heart Ventricles ; Male ; Myocardium

Objective To map the vision cortex of rats by dynamic manganese-enhanced functional magnetic resonance imaging and provide a method for researching the nervous function. Methods Six adult male Wistar rats were chosen and the process was divided into 4 continuous phases. No agent was injected into the rats in the first phase (5 min). Disrupting the BBB with marmitol and injecting manganese chloride were performed in the fight internal carotid artery (ICA) in the second phase (10 min). In the third phase (15 min), manganese chloride was administrated into theright ICA and vision stimulation was performed before the imaging process. The mixed liquor of manganese chloride and glutamate was injected into the rats in the forth phase (5 min). MRI was performed instantly after the handles in each phase. SPM and Matlab software were employed to help analyze the imaging data. Region-of-interest (ROI) was recorded to observe the stimulated regions and compare the signal intensity in the visual cortex. Results No specific enhanced region was found in the rat brain in the first and second phases. The right visual cortex was enhanced specifically on T1WI in the third phase. Many brain regions of the right hemisphere, the sites that agents was injected, were obviously enhanced in the forth 2008A1-E4011)phase. ROI analysis showed that the signal intensity in the third phrase (1.897±0.172) was significantly stronger as compared with that in the second phrase(1.549±0.163)(P<0.05). Conclusion The dynamic manganese-enhanced functional magnetic resonance imaging can analyze the functional activities of the vision cortex in rats and provide a new method for researching the function of the nervous system.

Objective To explore the effects and related mechanism of nifedipine on vascular inflammation induced by cuff placement. Methods Adult male C57BL/6J mice ( 10 to 12 weeks of age)were assigned to control ( no cuff placement without nifedipine ) , cuff placement ( cuff placement without ( cuff placement without nifedipine) and treatment (cuff placement with nifedipine 1 or 5 mg · kg-1 · d-1) groups. Activity of NF-κB in injured artery was measured 5 days after operation. MCP-1 expression and nuclear translocation of NF-κB were examined in injured artery 7 days after operation. Results DNA-binding activity of NF-κB was significantly increased in the injured artery 5 days after cuff placement which could be downregulated by nifedipine 5mg · kg-1 · d-1. MCP-1 mRNA expression in the injured arteries was increased 7 days after cuff placement and which could be significantly attenuated by nifedipine 5 mg · kg-1 · d-1. Cuff placement decreased the cytoplasmic level of p50, IκBα, IκBβ, and increased the nuclear level of p50. Nifedipine 5 mg· kg-1 · d-1 significantly attenuated these changes. Conclusion Our results suggest that high dose nifedipine could suppresse expression of MCP-1 induced by injured arteries via the inhibing NF-κB DNA binding activity, thereby attenuating vascular inflammation.

This study was aimed to investigate the aggregation of rehydrated-lyophilized platelets. The aggregation rate of fresh and rehydrated-lyophilized platelets were measured by using thrombin, ristocetin, ADP and collagen as inductors and APACT2 aggregameter; the effects of intra- and extra-cellular trehalose on maximum aggregation rate of rehydrated-lyophilized platelets were detected by using ADP as an inductor. The results showed that the aggregation rate of fresh platelets was all about 100%, while aggregation rate of rehydrated lyophilized platelets was (70.17 +/- 7.36)%, (15.3 +/- 2.81)%, (68.67 +/- 6.86)%, (64.67 +/- 11.6)% respectively, when the concentration of thrombin, ristocetin, ADP and collagen was 1 U/ml, 1.6 mg/ml, 20 micromol/L and 2 microg/ml. The maximum aggregation rates of rehydrated-lyophilized platelets in intra- and extra-cellular trehalose, extracellular trehalose and blank control groups were (66.0 +/- 4.69)%, (25.3 +/- 2.42)% and (11.5 +/- 1.87)% (P < 0.01), meanwhile there was significant difference of rehydrated-lyophilized platelet aggregation rate between intra- and extra-cellular trehalose and extracellular trehalose groups (P < 0.01). It is concluded that the concentrations of thrombin (1 U/ml), ristocetin (1.6 mg/ml), ADP (20 micromol/L) and collagen (2 microg/ml) are optimal for platelets aggregation tests, the internal and extracellular trehalose significantly enhance the aggregation of rehydrated-lyophilized platelets.
Blood Platelets ; cytology ; drug effects ; metabolism ; Blood Preservation ; methods ; Freeze Drying ; methods ; Humans ; Platelet Aggregation ; physiology

The aim of this research was to study the technology and methods of loading lyoprotectant-trehalose into cytoplasm of human platelets before lyophilization, to optimize experimental conditions of loading trehalose, to investigate the changes of platelets response to agonists and activation after incubation of platelets for 4 hours at 37 degrees C in the presence of lyoprotectant-trehalose, to protract the figures of loading efficiency and intracellular trehalose concentration versus incubation time, temperature and external trehalose concentration, to optimize loading parameters. The response of platelets to different agonists--thrombin, ADP, collagen and ristocetin were measured respectively by APACT2 aggregometer before and after loading trehalose into platelets; the expressions of CD62p and PAC-1 on platelet membranes in the presence and absence of reversible platelets activation inhibitors were measured by flow cytometry respectively before and after loading trehalose into cytoplasm of platelets. The results showed that the loading efficiency was linear to incubation time (2 hours later) and incubation temperature (rang from 30 degrees C to 40 degrees C), respectively. The loading efficiency almost reached 60% when the platelets were incubated at 37 degrees C for 4 hours. The intracellular trehalose concentration was higher with the increase of the extracellular trehalose concentration (< 50 mmol/L). Compared to untreated groups, the values of MPV and aggregation to different agonists in treated groups showed no significant difference, respectively (P > 0.01). After incubation of platelets for 4 hours, the expression of CD62p increased to some extent, however, the expression of CD62p decreased again when the reversible platelets activation inhibitor PGE-1 and adenosine were added to the incubation buffer. It is concluded that 37 degrees C, 4 hours and the extracellular trehalose concentration < 50 mmol/L are the optimal conditions for loading with trehalose. The processing of loading with trehalose before platelet lyophilization has no significant effects on response of platelets to agonists and activation.
Blood Platelets ; cytology ; drug effects ; metabolism ; Blood Preservation ; methods ; Cell Survival ; Cryopreservation ; methods ; Freeze Drying ; Humans ; Trehalose ; blood ; pharmacology

OBJECTIVETo explore effects of Yougui recipe (see text) and salmon calcitonin acetate in preventing osteolysis surrounding artificial prosthesis.

METHODSThirty-two SD male rats with weighted (250 +/- 20) g, aged 8 weeks, were randomly divided into four groups: blank group, model group, salmon calcitonin acetate group and Yougui recipe (see text) group, and 8 rats in each group. Blank group did not undergo any process, other 24 rats underwent anesthesia by chloral hydrate, their knee joints were exposed through medial patellar side,drilling from fermoral condyle nest to marrow cavity,high density of polythlene particles were injected into hole, titanium nail were put into, bone wax closed the window, then suturing step by step. After the molding, saline were used to gavaged in blank group and model group, Yougui recipe (see text) for Yougui recipe (see text) group, salmon calcitonin maximus injection for calcitonin group. After 10 weeks' mediation, rats were executed, and arterial blood and bilateral femoral organization were collected to biochemical, imaging morphology, tissue pathology and molecular biology detection.

RESULTSThe key gene expression of activiting osteoclast were inhibited in Yougui recipe (see text) group and calcitonin group. The level of OPG, Ca, ALP in Yougui recipe group were higher than calcitonin group (P<0.01); the content of RANKL were lower (P<0.01). There were no significance meaning in RANK, Trap5b, P between two groups.

CONCLUSIONBoth of Yougui recipe (see text) and calcitonin can slow and treat surrounding osteolysis of artificial joint prosthesis, and Yougui recipe (see text) has better effect in promoting bone formation. The effect of Yougui recipe (see text) in promoting bone formation, inhibiting osteoclasts to provide a new method to treating surrounding osteolysis of artificial joint prosthesis.

Animals ; Biomarkers ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Gene Expression Regulation ; drug effects ; Interleukin-1 ; genetics ; Interleukin-6 ; genetics ; Male ; Osteolysis ; etiology ; metabolism ; pathology ; prevention & control ; Prostheses and Implants ; adverse effects ; Rats ; Rats, Sprague-Dawley ; Tomography, X-Ray Computed ; Vacuolar Proton-Translocating ATPases ; genetics

This study was aimed to further optimize trehalose loading technique including loading temperature, loading time, loading solution and loading concentration of trehalose, based on the established parameters. Loading efficiency in plasma was compared with that in buffer at 37 degrees C; the curves of intracellular trehalose concentration versus loading time at 37 degrees C and 16 degrees C were measured; curves of mean platelet volume (MPV) versus loading time and loading concentration were investigated and compared. According to results obtained, the loaing time, loading temperature, loading solution and trehalose concentration were ascertained for high loading efficiency of trehalose into human platelet. The results showed that the loading efficiency in plasma was markedly higher than that in buffer at 37 degrees C, the loading efficiency in plasma at 37 degrees C was significantly higher than that at 16 degrees C and reached 19.51% after loading for 4 hours, but 6.16% at 16 degrees C. MPV at 16 degrees C was increased by 43.2% than that at 37 degrees C, but had no distinct changes with loading time and loading concentration. In loading at 37 degrees C, MPV increased with loading time and loading concentration positively. Loading time and loading concentration displayed synergetic effect on MPV. MPV increased with loading time and concentration while trehalose loading concentration was above 50 mmol/L. It is concluded that the optimization parameters of trehalose loading technique are 37 degrees C (temperature), 4 hours (leading time), plasma (loading solution), 50 mmol/L (feasible trehalose concentration). The trehalose concentration can be adjusted to meet the requirement of lyophilization.
Blood Platelets ; cytology ; drug effects ; metabolism ; Blood Preservation ; methods ; Cryopreservation ; methods ; Cryoprotective Agents ; metabolism ; pharmacology ; Dose-Response Relationship, Drug ; Freeze Drying ; Humans ; Trehalose ; metabolism ; pharmacology

The aim of this study was to search a procedure of platelet lyophilization and find a way of long-term storage of human platelets at normal temperature with smaller size and lighter weight, to be convenient to transport at long distance thus to meet the demands in accidents and war time. Human platelets were pretreated by freezing, the first and the second desiccation, and were added with reversible activation-inhibitors of platelets, DMSO and trehalose, then were rehydrated. At the same time, the recovery rate of platelets, platelet maximal aggregation induced by thrombin, coagulation of platelets, CD62p expression and PAC-1 expression were assayed. The results indicated that the recovery rate of the platelets was 56.29%. The platelet maximal aggregation induced by thrombin had no significant difference between lyophilized platelets and the fresh platelet-rich plasma (FPRP), but the aggregation of platelets induced by ADP or propyl gallate was decreased by 49.34% and 26.25%. Coagulation of the lyophilized platelets was not significantly different from FPRP. CD62p expression of the lyophilized platelets (42.36%) was higher than that in FPRP while PAC-1 expression was 2.12%. CD62p re-expression rate induced by thrombin was 50.88% and PAC-1 re-expression was 54.55%. It is concluded that the ability of recovered lyophilized platelets added with reversible activation-inhibitors, DMSO and trehalose to aggregate and coagulate has showed no significant difference as compared with FPRP. The reversible activation-inhibitors can decrease CD62p expression of lyophilized platelets, and may enhance their survival ability and prolongate survival time. Therefore the efficiency of lyophilizing platelets can be improved based on this freeze-drying procedure.
Blood Platelets ; cytology ; drug effects ; metabolism ; Blood Preservation ; methods ; Cell Survival ; Freeze Drying ; methods ; Humans ; Trehalose ; pharmacology

Objective To analyze the feasibility of local LINGO-1 polyclonal antibody administration for treatment of spinal cord injury in adult rats. Methods Twenty-four adult female SD rats were randomized into sham-operated group, rabbit IgG group and LINGO-1 antibody group. In the latter two groups, partial transaction of the T9 segment of the spinal cord was performed to completely sever the dorsal eorticospinal tract, followed immediately by administration of rabbit IgG and anti-LINGO polyclonal antibody via a mini-osmotic pump, respectively. At 3 and 28 days after the operation, the T8～10 segments of the spinal cord were harvested to prepare cryosections, and immunofluorescence staining was used to analyze the penetration of LINGO-1 polyclonal antibody into the spinal cord tissue and its specific binding to LINGO-1 molecules. Results In LINGO-1 antibody group, the presence of rabbit antibodies was detected at the injured sites of the spinal cord at 3 and 28 days after the operation. The mean immunofluorescence density was significantly lower in L1NGO-1 antibody group than in rabbit IgG group at 3 days after the operation (P<0.05). In rabbit IgG group, the mean immunofluorescence density for LINGO-1 in the crysections pre-treated with LINGO-1 polyclonal antibody was significantly lower than that in sections pre-treated with rabbit IgG(P<0.05). Conclusion Locally administered LINGO-1 polyclonal antibody can penetrate into the injured sites in the spinal cord in a wide time window and recognizes LINGO-1 molecule specifically, suggesting the feasibility of passive immunotherapy for spinal cord injury.