Objective To explore the effect of lysophosphatidic acid(LPA)on blood-brain barrier(BBB) permeability and its possible mechanism.Methods LPA or LPA+suramin(L+S)were stereotaxically injected into the right eaudate nucleus in SD rats in vivo.Evans blue(EB)was used to quantitatively measure the permeability of BBB at different time points.The expression of matrix metalloproteinase-9 was detected by immunohistochemistry technique.The pathological ultrastruetural changes of BBB were assessed by transmission electron microscopy.Results The BBB permeability began to increase after LPA administered into ipsilateral eaudate nucleus,and reached the peak at 24h.Then the permeability of BBB gradually lowered after 48h.In comparison with the same time points of control group,there were quite significant differences(P＜0.01).After L+S was injected,the change of BBB permeability had differences in comparison with those of LPA group in the same time points,(P＜0.05).MMP-9 positive cells were mainly vascular endothelial cells.The numbers of MMP-9 positive blood vessels grew at 6h in LPA group,and the expression of it reached maximum at 24h,then the number of it decreased at 48h,showing significant statistical differences in comparison with the L+S group(P＜0.01),It was observed microscopically that ultrastrueture of BBB of the LPA group was changed sharply,such as basement membrane roughed and fragmented,astroeyte end-feet swolled markedly and perivaseular space enlarged obviously.But there were no remarkable changes in BBB in L+S group.Conclusion LPA can induce increase of BBB permeability and its possible mechanism is the strong expression of MMP-9 protein produeted by endothelial cells through the mediation of LPA receptor,leading to degradation of basement membrane.

Objective To investigate the renal level of 1o-hydroxylase and the change of serum calcium in rats with severe acute pancreatitis,and their correlation.Methods Eighty male Wistar rats were randomly (random number) divided into two groups:sham operation group (SO group),severe acute pancreatitis group (SAP group),and each group was further randomly divided into 1 h,3 h,6 h,and 12 h subgroups (n =10).Severe acute pancreatitis model was made by retrograde infusion with 5％ sodium taurocholate solution into the biliopancreatic duct,rats were sacrificed at 1 h,3 h,6 h,and 12 h separately after modeling.The levels of serum amylase,serum calcium,serum urea nitrogen,serum creatinine,serum 1,25-dihydroxy vitamin D3 were measured,and the level of lα-hydroxylase protein in the kidney was determined with immunohistochemistry and Western blotting.The histopathologic changes of kidney tissue were observed under light microscope and the changes of the proximal tubular epithelial cell were observed under electron microscope.Results Compared with SO group,the levels of serum amylase,serum urea nitrogen,and serum creatinine were higher in SAP group,but the levels of serum calcium and 1,25-dihydroxy vitamin D3 decreased at 3,6,and 12 h,and the renal level of 1 α-hydroxylase also decreased at 3,6,and 12 h after modeling.In SAP group,the levels of serum calcium,serum 1,25-dihydroxy vitamin D3 and the renal level of 1 α-hydroxylase gradually decreased,and the renal level of 1 α-hydroxylase and the level of serum 1,25-dihydroxy vitamin D3 had positive correlation at 3 h,6 h,and 12 h (r =0.93,P ＜0.01; r=0.951,P ＜0.01; r =0.92,P ＜0.01; r =0.878,P ＜0.01),and the renal level of 1α-hydroxylase and the level of serum calcium had positive correlation at 3 h,6 h,and 12 h (r =0.975,P ＜0.01; r=0.946,P＜0.01; r=0.747,P＜0.01).Conclusions Intheearly course of SAP,the lowered activity of 1 α-hydroxylase may play an important role in the development of hypocalcemia.

Objective To investigate the perfusion characteristics of intraductal breast lesion by real-time gray-scale contrast ultrasound and to determine the value of real contrast ultrasound in the diagnosis of breast masses.Methods A total of 30 breast lumps by ultrasound contrast enhancement were observed from the enhanced level.An enhanced mode and enhanced border were observed when the lesion was clear.The perfusion characteristics were compared between the benign and malignant lesions.Results Thirty breast lumps include 17 benign lumps and 13 benign lumps by pathological operation.After injected with the microbubble contrast medium,all breast lumps enhanced to varied extent.And malignant lesions showed significant enhancement for more than 3 grade(69.2%,9/13).The radial enhancement around lesion were mainly observed in the malignant lesions (P＜0.05).Conclusion The microvascular perfusion of breast intraductal lesions can be clearly displayed by real-time gray-scale contrast-enhanced ultrasound.The feasibility of differentiation between benign and malignant lesions according to their perfusion characteristics appears to be promising.

Adult ; Bone Cysts, Aneurysmal ; diagnosis ; surgery ; Bone Neoplasms ; diagnosis ; surgery ; Chondroblastoma ; diagnosis ; surgery ; Female ; Humans ; Male ; Talus ; surgery ; Young Adult

Objective To investigate the sensitivity and variability factors that were assessed on the forced swimming test (FST) using BALB/C and Kunming mice. Methods The immobility time of FST was compared using Kunming and BALB/C mice in different experimental conditions including circadian rhythm ( day and night) ,gender and water temperature ( 12,22 and 32℃ ) . Results (①) The immobility time of BALB/C during the daytime( ( 142.42 ± 33.58) s) was significantly increased than that at night ( ( 104.89 ± 34.33 ) s). (② The immobility time of Kunming mice( (91.95 ± 40.32) s) was significantly decreased than that of BALB/C mice ( ( 142.42 ± 33.58 ) s). (③)The immobility time under the water temperature of 22 C ( ( 92.24 ± 25.81 ) s) was significant longer than that under the water temperature of 32C ( (60.72 ± 11.11 ) s). Conclusion BALB/C stain,male mice,daytime and water temperature of 22℃ should be chosen in the FST.

Objective To manufacture magnetic microbubbles with dual-response to ultrasound and magnetic fields.Methods Microbubbles of ultrasound contrast agent (ST68) based on a surfactant were prepared by the acoustic cavitation method.Fe3O4 magnetic nanoparticles with negative charge were synthesized using the polyol procedure.Magnetic microbubbles were generated by depositing polyethylenimine and Fe3O4 magnetic nanoparticles alternately onto the microbubbles using the layer-by-layer self-assembly.In vitro ultrasonography was performed on a silicone tube with/without magnetic microbubbles (3 × 108/ml) by a self-made device to observe the movement of magnetic microbubbles under the effects of magnetic field.In vivo imaging was performed on the kidney of New Zealand rabbits before and after the injection of magnetic microbubbles.Results The Fe3O4 nanoparticles carried a stable negative charge of (-24.6 ± 6.7) mV and more than 98％ of the particles were less than 8 μm in diameter,meeting the size requirement of an ultrasound contrast agent for intravenous administration.There was no echoic signal in the silicone tube before injection of magnetic microbubbles,but there were strong echoic signals after injection.After applying a magnetic field,the magnetic microbubbles moved along the direction of the magnetic flux.In vivo ultrasound imaging could not visualize the kidney before injection of magnetic microbubbles,but could remarkably visualize the kidney after injection.Conclusions The magnetic microbubbles exhibit favorable magnetic targeting and ultrasound contrast enhancement characteristics.Such properties may serve as the foundation to study their potential for simultaneous diagnosis and treatment in the future.

Objective To fabricate an ultrasound/fluorescence bi-modal contrast agent by encapsulating fluorescent quantum dots into polymeric ultrasound contrast agent microbubbles.Methods Polylactic acid (PLA,500 mg),(1R)-(+)-camphor (50 mg) and CdSe/ZnS quantum dots (0.5 ml,2.3 μmol/L)were dissolved or dispersed in dichloromethane (10 ml) to form in an organic phase.Ammonium carbonate solution and poly (vinyl alcohol) solution were employed as the internal and external water phase,respectively.The fluorescent microbubbles were generated using double emulsion solvent evaporation and lyophilization methods.The morphology and illumination were characterized by scanning electron microscopy (SEM) and fluorescence spectrophotometry.Synchronized contrast-enhanced ultrasound and fluorescence imaging was acquired by injecting fluorescent microbubbles into the silicone tube coupled to a self-made ultrasound/fluorescence imaging device.Ultrasound/fluorescence bi-modal in vivo imaging was acquired on the kidney of New Zealand rabbits and suckling mice.Results The fluorescent microbubbles were hollow spheres with an averaged diameter of (1.62 ± 1.47) μm.More than 99％ of these microbubbles were less than 8 μm in diameter,which meeted the size criteria for ultrasound contrast agents.The fluorescence emission peak of the microbubbles appeared at 632 nm,indicating that good luminescence properties of quantum dots were maintained.In vitro ultrasound/fluorescence imaging showed no echoic signal when the silicone tube was filled with saline,but there was a strong echo when filled with fluorescent microbubbles.The liquid column with fluorescent microbubbles emitted red luminescence under ultraviolet irradiation.The kidney of the rabbit was remarkably enhanced after the administration of fluorescent microbubbles.Bright fluorescence could be observed at the injection site of the suckling mice via subcutaneous injection.Conclusions A bi-modal but single contrast agent based on polymeric microbubbles has been successfully fabricated for the use of ultrasound and fluorescence imaging.It retains the good characteristics of both echogenicity and fluorescence,which complement each other in case of limitations imposed by uni-modal,single agents.

Objective To evaluate the protection of 3-aminobenzamide (3-AB),an inhibitor of Poly (ADP-ribose) polymerase (PARP),on severe acute pancreatitis associated adenohypophysis injury in rats.Method Forty Wistar rats were randomly divided into 4 groups:sham operation group (SO group,n=8),SAP group (n=12),3-AB pretreatment group (n =12),drug control group (n =8).The bilepancreatic duct was cannulated through the duodenum and SAP model was induced by a standardized pressure-controlled retrograde infusion of 5％ sodium taurocholate (0.1 ml/100 g) into the bile-pancreatic duct.In 3-AB group,3-AB (20 mg/kg) was administered via femoral vein 30 min prior to the operation;other procedures were identical to SAP group.In SO group,pancreas was flipped several times only.In drug control group,3-AB (20 mg/kg) was administered via femoral vein 30 min prior to the operation.Serum amylase,lipase were measured.Pancreas and pituitary tissue were taken for pathological examination under light microscope.PARP and NF-κB antibodies for adenohypophysis immunohistochemical stains.Adenohypophysis cell was observed under electronic microscope.Result Serum amylase,lipase and pancreas pathological scores were significantly higher in 3-AB group compared with SO group (P ＜ 0.05),but lower than that in SAP group (P ＜ 0.05).Adenohypophysis pathological injury was less severe in 3-AB group.Expressions of PARP and NF-κB in adenohypophysis cells were significantly higher in 3-AB group compared with SO group,but lower than that in SAP group (P ＜ 0.05).Ultrastructural change of thyrotroph cell was relieved in 3-AB group.No significant difference was observed between SO group and drug control group in PARP and NF-κB expression nor adenohypophysis pathological injury.Conclusions 3-AB exerts the protective effect against acute pancreatitis associated adenohypophysis injury by inhibition of PARP and NF-κB.

Objective To investigate the changes of poly-ADP-ribose polymerase (PARP) and NF kappa B (NF-κB) in adenohypophysis in rat model of severe acute pancreatitis (SAP),and their role in the mechanism of adenohypophysis injury in SAP.Methods Forty Wistar rats were randomly (random number) divided into 5 groups:the sham operation group (SO group,n =8),SAP 1 h,3 h,6 h and 12 h groups (n =8 in each group).SAP model was induced by retrograde injection of 5％ sodium taurocholate into the biliopancreatic duct.Serum levels of amylase,lipase and ascites were measured.After sacrifice of experiment rats,pancreas and adenohypophysis tissues were taken for pathological examination under light microscope.Adenohypophysis cells were observed under electronic microscopy as well.PARP and NF-κB expressions in adenohypophysis cell was studied by using immunohistochemisty assay.Results After modelling,serum levels of amylase,lipase and ascites in SAP group increased gradually,which were higher than those in SO group (P ＜ 0.05).Adenohypophysis cell swelling and partial necrosis were observed under light microscope.As the time prolonged,their nuclei became dark and pyknotic more and more,and the endoplasmic reticulum and mitochondrial swelling in adenohypophysis cells were observed under electronic microscopy.The expressions of PARP and NF-κB in SAP group increased gradually,which were higher than those in SO group.Conclusions Significant pathological and ultrastructural injuries were observed in adenohypophysis cells in severe acute pancreatitis.These changes might correlate with PARP and NF-κB signaling pathway.