Urotensin Ⅱ,a cycle peptide originally found from the fish,is the most potent vasoconstrictor.Urotensin Ⅱand its receptor have been found in central nerve system,cardiovascular system and other organs and tissues.It has been suggested that urotensin Ⅱplays important roles in the physiological and pathological procedures.Based on the evidences obtained from experiments,urotensin Ⅱmay become a new target for treating many diseases,particularly for cardiovascular diseases.

To screen potential hamster chymase 2 inhibitors, a high-throughput screening (HTS) model was established. Recombinant hamster chymase 2 with active form was cloned and expressed in E. coli. The HTS model with total volume of 50 microL in 384-well microplate was based on fluorescence analysis and was proved sensitive as well as specific (Z' = 0.84). A total of 40 080 samples (including 28 060 compounds and 12 020 natural products) were screened, and 613 samples with inhibition greater than 90% were selected for further rescreening. Finally, compounds J16647 and J16648 were identified with high inhibitory activity on chymase 2, and whose IC50 values were 0.823 and 0.690 micromol x L(-1), respectively.

Experimental studies on the myocardial ischemia and reperfusion injury in 16 anaethetized SDrats,of which,8 animals were pretreated with morphine(5 mg/kg,i.p.)for preventing of arrhyth-mias,were studied immunocytochemically with anti-muscle actin specific monoclonal antibody (HHF_(35)),8 shan-operated rats were used as control.With HHF_(35) ABC immunocytochemical method,the area of myocardial ischemia and reperfusion injury(without morphine)showed decrease or ab-sence of staining,large areas of staining loss were also seen.In the group with morphine,only smallfoci of staining absence were shown.The myocardium in control animals showed evenly positive stain-ing.No change were seen with HE staining in all groups.The results obtained with HHF_(35) stainingsupport its important value in studying on myocardic reperfusion injury,and indicated that the degreeof myocardic damage may be relative to the arrhythmias in myocardial reperfusion injury.

Objective To investigate the efficacy of continuous veno-venuous hemodialysis/filtration(CVVHD/F) for the treatment of multiple organ dysfunction syndrome(MODS)caused by severe infection and to explore the mechanism in children.Method Nineteen cases of pediatric septic shock with MODS were treated with CVVHD/F in Children's Hospital Affiliated to Shanghai Jiaotong University from December 2002 to November 2007.The clinical data were studied including mortality rate,serum electrolytes,arterial partial pressure of oxygen (PO2),artery partial pressure of carbon dioxide(PCO2),FiO2/PO2,urine output,blood pressure,doses of vasoactive agents,Cr,BUN,etc.Results Cannulation and CVVHD/F were well performed in a total of 19 cases,with median age 33.4±36.5 months(from 3 months to 8 years) ,with their gender ratio of male(13 cases)to female (6 cases) to be 68.4% and 31.6%.The mean pediatric crifcal illness score(PCIS) was 69.1±10.4 and Median Pediatric Risk of Mortality score(PRMS Ⅲ)12.66±7.85,respectively.The duration of CWHD/F was 92 hours(ranged from 16 hours to480 hours).FiO2/PO2,PCO2,and PO2 were iraproved significantly after 12 to 24 hours CVVHD/F in patients with acute respiratory distress syndrome(ARDS) or lung edema (P<0.05).The concentrations of serum kalium,natrinm and HCO3- level resumed to well-balanced in 24 hours (P<0.05).The serum Cr and BUN were decreased to normal range(P<0.05).The mortality rate was 63.2%.Conclusions CVVHD/F was effective for treatment of septic shock with MODS in pediatric by improving oxygenation,maintaining normal serum electrolytes,conecting metabolic acidosis,increasing the tissue perfusion and eliminating the serum Cr and BUN.

BACKGROUND: Fibrin glue has been demonstrated to function as a kind of biomaterial with high quality. It has been used in nerve tissue engineering and proved to be a kind of scaffold for some cells.OBJECTIVE: To explore the biocompatibility of fibrin glue and olfactory ensheathing cells (OECs).DESIGN, TIME AND SETTING: An in vitro control trial based on cytology was performed at the Institute of Neurobiology,Nantong University from August 2007 to February 2008.MATERIALS: Fibrin glue was made of fibrin and catalyst, and OECs derived from rats' olfactory bulb were normally primary-cultured.METHODS: OECs were divided into control (OECs clone spheres were cultured alone) and in fibrin glue (OECs clone spheres were cultured and combined with fibrin glue) groups. After 1 week of culture, the proliferation of OECs were observed by convert microscope and detected by S-100 immunofluorescence histochemical staining.MAIN OUTCOME M EASURES: OECs morphology, cell count, the area of the cell bodies and the perimeter of the cell were determined.RESULTS: OECs could survive, migrate in fibrin glue, and float in the fibrin glue in the lower layer. After 7 days of incubation, cell body exhibited fusiform or triangle, predominantly bipolar or bipolar. The number of the S-100 positive cells was more, and cell bodies were larger in fibrin glue group than control group (P < 0.05). However, there was no obvious difference between two groups in cell perimeter (P > 0.05).CONCLUSION: Fibrin glue has good biocompatibility with OECs, and OECs can survive and migrate in fibrin glue.

The present research was aimed to explore the biocompatibility of IKVAV self-assembling peptide nanofiber scaffold with olfactory ensheathing cells (OECs) of rats. The OECs were seeded onto the surface of coverslips covered with IKVAV self-assembling peptide nanofiber scaffold hydrogel (2D culture system), and implanted within IKVAV self-assembling peptide nanofiber scaffold hydrogel (3D culture system), respectively. The adhesion, viability of OECs were observed with inverted microscope. Then the characteristics for survival and adhesion of cells by image processing were observed, and statistical analysis on the number of S-100 positive cell, the area of the cell bodies and the perimeter of the cell and MTT method were carried out. It was found that the OECs could survive and migrate in IKVAV self-assembling peptide nanofiber scaffold. The result of the cell MTT exam, of the shape and quantity of cells had no significant difference compared to those of the OECs cultured with poly-L-lysine (PLL). It has been proved that IKVAV self-assembling peptide nanofiber scaffold has good biocompatibility with rat OECs.
Animals ; Animals, Newborn ; Biocompatible Materials ; chemistry ; Cell Proliferation ; Cells, Cultured ; Hydrogel, Polyethylene Glycol Dimethacrylate ; chemistry ; Laminin ; chemistry ; Nanofibers ; chemistry ; Olfactory Bulb ; cytology ; drug effects ; Peptide Fragments ; chemistry ; Rats ; Rats, Sprague-Dawley ; Tissue Engineering ; methods ; Tissue Scaffolds ; chemistry

Objective To evaluate a new biodegradable inferior vena cava(IVC) filter by in vitro and in vivo experiments.Methods The biodegradable filter consisted of metal stent and absorbable suture,which acts as a degradable deformable switch of the filter.When the suture of the proximal metal filter degrades and lost its tension,the metal filter will change into a stent.In vitro experiments were performed to test the influence of the sterilization method and the tension of absorbable sutures to the filter's selfconvertible time,animal experiments were conducted to evaluate the effectiveness of emboli capture and to observe the in vivo self-convertible time of the filter.IVC stenosis,pulmonary embolus and intimal hyperplasia and inflammation were evaluated.Results Ethylene oxide sterilization did not have an adverse effect on the self-convertible time of the filters.The tension did not affect the degradation rates of the suture significantly.In animal experiments,the VCFs were successfully implanted via femoral vein approach.Not any tilt,migration or structural damage of the filters was found during the follow-up time.Postoperative fluoroscopy and autopsy confirmed that there was no stenosis or thrombosis,IVC perforation.After implantation large thrombi were captured.The sutures degraded and filters transformed into stents in 3 weeks.After 90 days IVC was patent with mild intimal hyperplasia and no thrombosis.Conclusions This study demonstrated the effectiveness and safety of the biodegradable filter we designed.

Objective To explore the experimental methods and conditions of 131I-labeled anti-epidermal growth factor receptor (EGFR) vⅢ preparation,and to evaluate the targeting distribution of 131I-Anti-EGFRvⅢ in malignant glioma-loading nude mice.Methods The 131I labeling on anti-EGFRvⅢ was performed by Iodogen method.The labeling rate was determined after separation and purification and paper chromatography was used for the determination of radioactive chemical purity.Twenty-eight U87-EGFRvⅢ malignant glioma-loading nude mice with glioma average diameter of 10-15 mm were chosen and randomly divided into group of 131I-Anti-EGFRvⅢ intravenous injection,group of Na131I intravenous injection,group of 131I-Anti-EGFRvⅢ intratumor injection and group of Na131I intratumor injection;7.5 MBq/0.1 mL labeled products with 131I-Anti-EGFRvⅢ or Na131I were injected in the veins or the tumors to observe the changes of the radioactivity distribution of malignant glioma-loading nude mice with SPECT imaging.Results The rate of 131I-labeled anti-EGFRvⅢ was (68.12±6.19)％,and the immediate rate of radiochemieal purity was (95.12±0.59)％,and (87.78 ±5.35)％ in room temperature and (85.12±3.58)％ in 37 ℃ serum placed for 24 h.SPECT scan showed that the tumor site had significantly stronger imaging than the thyroid gland with the labeled products either by intravenous or intratumor injection.Conclusions It is applicable to the 131I-labeled Anti-EGFRvⅢ with Iodogen method.131I-Anti-EGFRvⅢ has good radiation chemical purity and stability in vitro and in vivo,and could be combined with tumor tissue specificity.

Objective:To investigate the effects of high-energy shock and vibration on cortex injury and peripheral blood immune cells in goats.Methods:Seventeen Boer goats without gender preference were selected. By using random number tables, the goats were divided into normal control group ( n=5) and shock and vibration injury group ( n=12). The goats in the normal control group were anatomized routinely and their brain was collected after being sacrificed without any other treatment. The goats in the high-energy shock and vibration model group were placed on a loading table (part of the BY10-100 instant shock and vibration simulation platform) in a restrained state, and made into a high-energy shock and vibration injury model induced by a vertical impact waveform generator. The intravenous blood samples were taken from the goats in the shock and vibration injury group before and at 0, 3, 6 and 24 hours after injury.Then, the goats were sacrificed and the following procedures were the same as the normal control group. At 24 hours after injury, the brain injury and the histopathological changes of the cerebral cortex in the normal control group and shock and vibration injury group were observed by gross pathological and anatomical examination and HE staining. The mRNA expression of zonula occludens 1 (ZO-1), tight junction protein 5 (Claudin-5), glial fibrillary acidic protein (GFAP), ionized calcium binding adaptor molecule 1 (IBA-1), interleukin (IL)-1β, IL-6 and cluster of differentiation antigen 177 (CD177) of the cerebral cortex in the normal control group and shock and vibration injury group were measured through fluorogenic quantitative polymerase chain reaction. The expression of ZO-1 and Claudin-5 proteins of the cerebral cortex in the normal control group and shock and vibration injury group were detected by Western blotting. Hematology analyzer and coagulation analyzer were used to detect white blood cell count, neutrocyte, lymphocyte, monocyte, prothrombin time 1 (PT-1), prothrombin time international normalized ratio (PT-INR), activated partial thromboplastin time (APTT), thrombin time (TT), prothrombin activity (PTA) and fibrinogen (FIB) levels in goats of the shock and vibration injury group before and at 0, 3, 6 and 24 hours after injury, respectively. Results:At 24 hours after injury, no visible contusion or necrosis was found in goat brain tissue in the shock and vibration injury group; the cerebral micro-vessels presented with a local dilation, hyperemia, edema, aggregation of inflammatory cells, disruption of vessel walls and leakage of red blood cells. These changes were not observed in the normal control group. In the shock and vibration injury group, ZO-1 and Claudin-5 mRNA expressions in the cerebral cortex were 0.25±0.10 and 0.09(0.04, 0.44) respectively, which were significantly lower than those of the normal control group [1.00±0.15 and 0.99(0.80, 1.20)]; GFAP, IBA-1, IL-1β, IL-6 and CD177 mRNA expression levels were 4.40(3.88, 6.75), 2.60±1.07, 3.04±0.51, 2.71±0.45 and 2.93±0.62 respectively, which were significantly higher than those of the normal control group [1.00(0.78, 1.22), 1.00±0.37, 1.00±0.27, 1.00±0.57 and 1.00±0.35]; ZO-1 and Claudin-5 protein expression levels were 0.41±0.06 and 0.42±0.11 respectively, which were significantly lower than those of the normal control group (1.08±0.12 and 0.91±0.23) (all P<0.01). In the shock and vibration injury group, the levels of white blood count, neutrocyte, and lymphocyte in peripheral blood were (13.7±3.3)×10 9/L, (35.3±14.8)% and (57.2±15.1)% respectively before injury, (19.4±3.1)×10 9/L, (60.5±12.5)% and (33.6±14.2)% respectively at 3 hours after injury, and (20.6±3.6)×10 9/L, (63.6±13.0)% and (30.9±15.0)% respectively at 6 hours after injury. By contrast, the levels of white blood count and neutrocyte were significantly increased but the level of lymphocyte was significantly decreased at 3 and 6 hours after injury ( P<0.05 or 0.01); the levels of the above indicators showed no significant changes at 0 and 24 hours after injury (all P>0.05); the level of monocyte did not change significantly at all time points before and after injury (all P>0.05). The levels of PT-1, PT-INR, APTT, TT, PTA and FIB in the shock and vibration injury group did not change significantly at each time point before and after injury (all P>0.05). Conclusion:Cerebral cortex microvascular injury and disruption of blood-brain barrier can be initiated in the early stage of high-energy shock and vibration injury in goats, accompanied by the presence of central and peripheral inflammatory response.

Mutations or inactivation of parkin, an E3 ubiquitin ligase, are associated with familial form or sporadic Parkinson's disease (PD), respectively, which manifested with the selective vulnerability of neuronal cells in substantia nigra (SN) and striatum (STR) regions. However, the underlying molecular mechanism linking parkin with the etiology of PD remains elusive. Here we report that p62, a critical regulator for protein quality control, inclusion body formation, selective autophagy and diverse signaling pathways, is a new substrate of parkin. P62 levels were increased in the SN and STR regions, but not in other brain regions in parkin knockout mice. Parkin directly interacts with and ubiquitinates p62 at the K13 to promote proteasomal degradation of p62 even in the absence of ATG5. Pathogenic mutations, knockdown of parkin or mutation of p62 at K13 prevented the degradation of p62. We further showed that parkin deficiency mice have pronounced loss of tyrosine hydroxylase positive neurons and have worse performance in motor test when treated with 6-hydroxydopamine hydrochloride in aged mice. These results suggest that, in addition to their critical role in regulating autophagy, p62 are subjected to parkin mediated proteasomal degradation and implicate that the dysregulation of parkin/p62 axis may involve in the selective vulnerability of neuronal cells during the onset of PD pathogenesis.
Adaptor Proteins, Signal Transducing ; chemistry ; metabolism ; Animals ; HEK293 Cells ; Heat-Shock Proteins ; chemistry ; metabolism ; Humans ; Lysine ; metabolism ; Mice ; Neurons ; metabolism ; pathology ; Oxidopamine ; pharmacology ; Parkinson Disease ; metabolism ; pathology ; Proteasome Endopeptidase Complex ; metabolism ; Protein Stability ; Proteolysis ; drug effects ; Sequestosome-1 Protein ; Ubiquitin-Protein Ligases ; metabolism ; Ubiquitination ; drug effects