BACKGROUND: In animal experiments, ultrasound-mediated microbubbles can promote the homing of transplanted stem cells to the ischemic area, enhance angiogenesis and small arterial formation, improve local blood flow in the ischemic myocardium and restore myocardial contractility.OBJECTIVE: To investigate the effect of ultrasound-mediated microbubbles on intravenously transplanted bone marrow mesenchymal stem cell (BMSC) homing and the therapeutic efficiency on ischemic stroke. METHODS: A middle cerebral artery occlusion (MCAO) model was induced by plug wire preparation. At 72 hours after MCAO, model rats were randomized into four groups: PBS group (n=15), BMSCs group (n=18), ultrasound+BMSCs group (n=18), ultrasound+microbubble+BMSCs group (n=18). Corresponding treatment was done in each group: 2 mL of PBS was injected via tail vein in the PBS group; about 3×106 BMSCs diluted by 2 mL of PBS were injected via tail vein slowly in the BMSCs group; after skull ultrasound radiation (1 MHz, 2 W/cm2) for 120 seconds, BMSCs were injected via tail vein slowly in the ultrasound+BMSCs group; the same process as the ultrasound+BMSCs group was done following intravenous injection of 0.1 mL/kg microbubbles in the ultrasound+microbubble+BMSCs group.RESULTS AND CONCLUSION: (1) Forty-eight hours after BMSCs transplantation, the BMSCs homing rate in the brain was significantly higher in the ultrasound+microbubble+BMSCs group than the other two groups (P < 0.05). (2) Twenty-eight days after MCAO, nerve damage was significantly milder in the ultrasound+microbubble+BMSCs group than the other two groups (P < 0.05). (3) Seven days after transplantation, the water content in the brain tissue was significantly lower in the ultrasound+microbubble+BMSCs group than the other two groups (P < 0.05). (4) Seven days after transplantation, the cerebral infarction volume was significantly reduced in the ultrasound+microbubble+BMSCs group compared with the other two groups (P < 0.05). To conclude, ultrasound-mediated microbubbles can enhance the homing effect of intravenously transplanted BMSCs, reduce cerebral edema and cerebral infarction volume, improve the neurological function, and increase the therapeutic effect of BMSCs transplantation on ischemic stroke.

Objective Toinvestigatetheclinicalsignificanceofplasmamatrixmetalloproteinase9 (MMP-9)intheformationofdelayedcerebraledemaafterintracerebralhemorrhage.Methods The clinical data of 107 patients with spontaneous intracerebral hemorrhage treated with conservative medical treatment were analyzed retrospectively. According to the clinical features and imaging changes,they were divided into either a delayed cerebral edema group (case group n=39)or a non-delayed cerebral edema group (control group n =68 ). The plasma MMP-9 level was detected with enzyme-linked immunosorbent assay within 24 h after onset. The patients performed head CT scan again at day 7 and 14 after admission. The changes of hematoma and edema volume were detected. All the possible factors associated with the formation of delayed cerebral edema were firstly analyzed by the univariate analysis. Univariate analysis showed that the variables with significant differences were enrolled into multiple logistic regression analysis. Results TheplasmaMMP-9levelofthedelayedbrainedemagroupwassignificantlyhigherthanthatof the control group,they were 189 ± 51 and 118 ± 27 mg/L respectively (P<0. 01). The result of univariate analysis showed that age,history of smoking,blood glucose level,baseline hematoma volume,and National Institute of Health stroke scale (NIHSS )score on admission might be associated with the formation of delayed cerebral edema after intracerebral hemorrhage. Logistic regression analysis showed that MMP-9 level (OR,9. 745,95%CI 6. 754-15. 466,P<0. 01),baseline hematoma volume (OR,2. 411,95%CI 1. 190-2. 728,P =0. 018),blood glucose level on admission (OR,1. 327,95%CI 1. 133 -1. 850,P =0.004),and NIHSS score (OR,1. 867,95%CI 1. 272-2. 364,P=0. 020)were the independent risk factorsfortheformationofdelayedcerebraledemaafterintracerebralhemorrhage.Conclusion Theamount of bleeding,NIHSS score,and hyperglycemia are the risk factors for the formation of delayed cerebral edema in patients with spontaneous intracerebral hemorrhage,while high MMP-9 level on admission indicated that the risk of the formation of delayed cerebral edema is high.

ObjectiveToinvestigatetheriskfactorsofepilepticseizuresanditseffectonclinical outcome in patients w ith cerebral venous sinus thrombosis (CVST). Methods The patients w ith CVST w ere enrol ed retrospectively. The risk factors, clinical manifestations, and imaging data w ere col ected. The data of an epileptic seizure group and a non-epileptic seizure group w ere compared. Results A total of 69 patients with CVST were enroled, including 32 (46.38%) secondary epileptic seizures. In the aspect of clinical manifestations, more patients show ed hemiplegia in the epileptic seizure group (37.50%vs.15.63%; χ2 =5.240, P=0.020). Imaging examination show ed that more patients in the epileptic seizure group presented w ith bleeding ( 29.41%vs. 10.81%; χ2 = 3.818, P= 0.047 ), more lesion involving frontal lobe (31.25%vs.10.81%; χ2 =5.008, P=0.023), and temporal lobe (43.75%vs.8.11%; χ2 =7.318, P=0.005), and the thrombosis sites w ere more common in the superior sagittal sinuses (65.63%vs.40.54%;χ2 =4.264, P=0.036). Multivariate logistic regression analysis show ed that focal neurological deficits (odds ratio 5.167, 95% confidence interval 1.993-15.764; P=0.004) and superior sagittal sinus thrombosis (odds ratio 0.126, 95% confidence interval 0.042-0.370; P=0.039) w ere the independent risk factors for patients w ith secondary epileptic seizures. There w ere no significant differences in hospital mortality (6.25%vs.2.7%; χ2 =0.512, P=0.469 ) and 90 day 90-day ful recovery rate ( defined as Barthel Index >60) (81.25%vs.86.47%; χ2 =0.346, P=0.793) betw een the epileptic seizure group and the non-epileptic seizure group. Conclusions Focal neurologic deficits and superior sagittal sinus thrombosis are the independent risk factors for secondary epileptic seizures, how ever, secondary epileptic seizures is not associ-ated w ith in-hospital mortality risk and 90-day clinical outcomes in patients w ith CVST.

Objective To assess the safety of early subcutaneous injection of a low-dose low molecular weight heparin (LMWH) nadroparin for prevention of deep vein thrombosis (DVT) in patients with spontaneous intracerebral hemorrhage (sICH).Methods The patients with sICH who early using nadroparin or lower limb intermittent pneumatic compression (IPC) for prevention of DVT were enrolled.A nadroparin group continuously injected nadroparin 0.4 ml/d subcutaneously for 10 days at day 4 after admission and an IPC group used lower limb IPC.Head CT was reexamined and hematoma volume changes were evaluated at day 3,5,and 14 after admission.The hemorrhagic events during the course of treatment were documented,and the lower limb DVT was examined by color Doppler sonography.Results A total of 94 patients with acute sICH (n =41 in the nadroparin group,n =53 in the IPC group) who early use of nadroparin or IPC for prevention of DVT were enrolled.Fourteen patients had lower limb DVT,5 (12.2％) of them were in the nadroparin group and 9 (17.0％) of them were in the IPC group.However,there was no significant difference in the incidence of DVT between the two groups (x2 =0.418; P =0.518).During the treatment,no patient experienced increased intracranial hematoma and rebleeding.Conclusion Early subcutaneous injection of low-dose nadroparin for the prevention of DVT in patients with sICH is safe.

Objective To investigate immunophenotypic characteristics of uterine natural killer (uNK)cells and helper T cell 1/helper T cell 2(Th1/Th2)immunity in third trimester decidua in preeclampsia.Methods The proportions of uNK cell subsets,expression of CD_(69)and CD_(94)on uNK cells and Th1/Th2 immunity in decidua were determined in 20 cases of preeclampsia patients and 11 cases of normal term pregnancies by flow cytometric analysis.Results The percentage of CD_(56)~(bright)CD_(16)~-uNK cell subset in preeclampsia patients and the controls was(17.3?11.1)% vs(17.9?16.8)%,that of CD_(56)~(dim) CD_(16)~+uNK cell subset was(16.3?8.7)% vs(16.2?8.8)%;that of CD_(56)~+CD_(69)~+uNK cells was(37.9 ?18.9)% vs(36.8?19.7)%,that of CD_(56)~+CD_(94)~+uNK cells was(34.9?15.2)% vs(32.7?16.2)% and the ratio of CD_(56)~+CD_(69)~+/CD_(56)~+CD_(94)~+was 1.1?0.2,1.2?0.6.No statistical difference was shown in the above values between the preeclampsia patients and controls.The percentage of cytoto xic T cell(Tc)2 cells was significantly lower in the decidua of preeclampsia patients [(3.0?1.0)% vs(4.3?0.9)%,P= 0.001 ],and the ratio of Tc1/Tc2 in preeclampsia patients was significantly higher than that of normal term pregnancies(17.8?3.4 vs 11.8?4.6;P=0.001);the ratio of Th1/Th2 was increased(15.1?2.4 vs 13.2?3.1;P=0.06).Conclusions The immunophenotypie characteristics of uNK cells do not present any significant change in preeclampsia patients.Owing to Tc2 cell decrease,the Th1/Th2 immunity shifts to Th1 type immunity in the decidua,which might contribute to the pathogenesis of preeclampsia.

BACKGROUND:Animal studies have indicated ultrasound-mediated microbubbles can significantly enhance the effect of stem cel transplantation to treat ischemic diseases. But its mechanism is stil unknown.
OBJECTIVE:To explore the mechanism of ultrasound-mediated microbubbles to significantly enhance the effect of stem cel transplantation in the treatment of ischemic diseases.
METHODS:Bone marrow mesenchymal stem cel s and vascular endothelial cel s of rats were cultured in vitro, and then randomized to three groups:control group with no intervention, ultrasound group exposed to ultrasound at 1 MHz, 1 W/cm2 for 90 seconds, and ultrasound-mediated microbubble group treated with 5μL liposomes ultrasound microbubbles containing fluorocarbon gases (about 2×1011/L) and ultrasound exposure at 1 MHz, 1 W/cm2 for 90 seconds.
RESULTS AND CONCLUSION:Compared to the control group, ultrasound-mediated microbubbles significantly increased expressions of vascular endothelial growth factor and stromal cel-derived factor 1 in the supernatant of
vascular endothelial cel s (P<0.05);ultrasound had no effect on the expression of vascular endothelial growth factor, but decreased the level of stromal cel-derived factor 1 (P<0.01). Ultrasound-mediated microbubbles and the ultrasound alone could significantly enhance the CXCR4 gene expression in bone marrow mesenchymal stem cel s as compared with the control group (P<0.01), but there was no difference between the ultrasound-mediated microbubble group and the ultrasound group (P>0.05). These findings suggest that 1 W/cm2 ultrasound-mediated microbubbles can promote vascular endothelial growth factor and stromal cel-derived factor 1 secretion by vascular endothelia cel s, and meanwhile promote CXCR4 gene expression in bone marrow mesenchymal stem cel s. This may be the mechanism of the ultrasound-mediated microbubbles enhancing homing effect of transplanted stem cel s.

AIM:To remark the effect of Qingguang'an Ⅱ on expression of PAX6, Ngn1, and Ngn2 mRNA of rats with chronic high intraocular pressure.METHODS:Totally 40 male SD rats were randomly divided into 6 groups, that was:A:blank group, B:model group, C:Qingguang'an Ⅱ low dose group, D:Qingguang'an Ⅱ moderate dose group, E:Qingguang'an Ⅱ high dose group, F:Yimaikang disket group.B, C, D, E, F groups of experimental rats were established the model of chronic high intraocular pressure (IOP) by cauterizing of superficial scleral vein.Animal model was established successfully by using monitoring IOP consistently keep above 25mmHg for 8wk as cut-off criterion.Tissues of Eyes were obtained after intragastric administration for 2wk and 4wk.The expressions of PAX6, Ngn1, and Ngn2 mRNA were investigated by Real-time PCR.RESULTS:At the time-point of 2wk, PAX6, Ngn1, and Ngn2 mRNA in group B were statistically expressed in lower level comparing with other groups (P<0.05).Moreover, at the time-point of 4wk, PAX6, Ngn1, and Ngn2 mRNA in group C, D and E were statistically expressed in higher level comparing with group F (P<0.05).Besides, PAX6, Ngn1, and Ngn2 mRNA in group C and D were statistically expressed in lower level comparing with group E (P<0.05).PAX6, Ngn1, and Ngn2 mRNA in group C and D were expressed in similar level(P>0.05).CONCLUSION:In summar, Qingguang'an Ⅱ and Yimaikang disket can remarkably increase the expressions of PAX6, Ngn1, and Ngn2, which suggest protecting the optic nerve of rats caused by chronic high IOP.What's more, this study indicated that, in the protection of optic nerve of rats with chronic high IOP, the high dose of Qingguang'an Ⅱ at the time-point of 4wk was the better choice.

OBJECTIVETo observe the expression of extracellular matrix metalloproteinase inducer (EMMPRIN) in the unstable plaque of patients with acute coronary syndrome (ACS), and the impact of leukotriene B4 (LTB4) on the EMMPRIN expression in macrophages.

METHODSThe EMMPRIN expression was detected by immunohistochemistry in 11 unstable plaques from patients with ACS. Protein expression of EMMPRIN was evaluated by Western blot on macrophages differentiated from THP-1 which were stimulated with LTB4 in the absence or presence of LTB4 antagonist U75302. There are 8 study groups: 1-THP-1, 2-8-the macrophages derived from THP-1, 2-6-macrophages were stimulated by LTB4 (0, 10(-10), 10(-9), 10(-8) and 10(-7) mol/L) for 24 h, 7-8-the macrophages were pretreated by 10(-6) mol/L or 10(-7) mol/L U75302 2 h before the LTB4 (10(-7) mol/L) stimulation.

RESULTSAbundant EMMPRIN expression was detected in macrophages and smooth muscle cells of unstable plaques from ACS patients. As to the THP-1 derived macrophages, EMMPRIN expression was significantly upregulated in a concentration-dependent manner in LTB4 stimulated groups, which was significantly higher in group 3-6 than in the THP-1 group (group 1) and macrophages group (group 2) (all P < 0.05) and pretreatment with U75302 significantly reduced the LTB4 induced upregulation of EMMPRIN in a dose-dependent manner (P < 0.05).

CONCLUSIONEMMPRIN expression is enhanced in macrophages and smooth muscle cells on unstable coronary artery plaques from ACS patients. LTB4 could stimulate EMMPRIN expression on THP-1 derived macrophages suggesting that LTB4 and EMMPRIN might be both involved in the formation and progression of unstable plaques, future studies are warranted to explore if LTB4 and EMMPRIN antagonists are effective or not for treating patients with ACS.

Acute Coronary Syndrome ; metabolism ; pathology ; Basigin ; metabolism ; Cell Line ; Humans ; Leukotriene B4 ; metabolism ; pharmacology ; Macrophages ; drug effects ; metabolism ; Myocytes, Smooth Muscle ; drug effects ; metabolism ; Plaque, Atherosclerotic ; metabolism

OBJECTIVETo evaluate the clinical effect of one stage anterior and posterior fusion and posterior fixation for the treatment of thoracic and lumbar spinal tuberculosis.

METHODSFrom March 2003 to December 2006, one stage anterior and posterior fusion and posterior fixation were performed to treat 23 patients who suffered thoracic and lumbar spinal tuberculosis. There were 15 males and 8 females with an average of 37.6 years (17-61 years). 4 cases' tuberculose focus were in thoracic vertebra, 8 cases in thoracolumbar, 11 cases in lumbar.

RESULTSThe average follow up period was 28.7 months (9-40 months). The symptoms of all patients had primarily relieved and the patients can ambulate at 2-3 weeks after treatment. At the 6th after operation, the X-ray showed interbody fusion. Frankel grading of 16 patients with incomplete paraplegia were improved averagely 1.62 grades. The major complications including 2 cases of temporary sinus formation, 1 case of fixtor breaking and 1 case of recurring (owing to an inadequate postoperative chemotherapy).

CONCLUSIONOne stage anterior and posterior fusion and posterior fixation can effectually resect focus, rebuild stability of spine, promote interbody fusion and recovery of incomplete paraplegia in treating thoracic and lumbar spinal tuberculosis.

Adolescent ; Adult ; Female ; Fracture Fixation, Internal ; Humans ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Postoperative Complications ; Spinal Fusion ; Thoracic Vertebrae ; surgery ; Treatment Outcome ; Tuberculosis, Spinal ; surgery ; Young Adult

OBJECTIVETo establish the two-dimensional electrophoresis profiles with high resolution and reproducibility from the brain tissues of mice with human cytomegalovirus (HCMV) infection.

METHODSForty Kunming mice were randomized into HCMV infection group (n=20) with HCMVAD(169) injection and control group (n=20) with saline injection in the brain. Thirty days after the injections, the brain tissue of the mice were taken and the protein fractions were isolated by two-dimensional gel electrophoresis (2-DE). Image Master 2D software was used to identify the differentially expressed proteins, and the peptide mass fingerprint (PMF) data were obtained for identification of the differential protein spots via database searching. Western blotting was performed to verify the expressions of some of the differential proteins.

RESULTS AND CONCLUSIONThe 2-D maps of the brain tissues with high Well resolution and reproducibility were obtained. Some of the differentially expressed proteins identified by mass spectrometry (MS) matched their counterparts in the SWISS-2DPAGE database. Western blotting analyses verified the differential expression of the individual proteins. These data can be of value for studying the diagnosis, pathogenesis and effective therapeutic targets of the disease.

Animals ; Animals, Newborn ; Brain ; metabolism ; virology ; Cytomegalovirus Infections ; metabolism ; Electrophoresis, Gel, Two-Dimensional ; Mice ; Proteins ; analysis ; Proteome ; analysis ; Proteomics ; methods ; Software