Aim To detect the expression of miRNA-99b and mTOR in glioma tissues and to investigate the effect of miRNA-99b on the cell invasion ability of hu-man glioma cell line U251. Methods The expres-sions of miRNA-99b and mTOR mRNA in glioma tis-sues and normal brain tissues were detected by real-time PCR. After co-transfection with miRNA-99b mim-ics and wild or mutation type mTOR 3′UTR recombina-tion vector,the specific binding ability of miRNA-99b to 3′-UTR in mTOR gene was examined by luciferase gene reporter system. The expression levels of miRNA-99b,mTOR mRNA and mTOR protein in glioma cell line U251 after transfected with miRNA-99b mimics were measured by real-time PCR and Western blot,re-spectively. The cell invasion was measured by Tran-swell assay. The changes of mTOR and miRNA-99b expression levels in U251 cells after transfection with mTOR PsiCHECK were detected by real-time PCR. The correlation between the expression of miRNA-99b and prognosis was analyzed statistically. Results The expressed level of miRNA-99b was lower, and the ex-pression level of mTOR was higher in the glioma tissues than that in the normal brain tissues. The expression of miRNA-99b was up-regulated, and the expressions of mTOR mRNA and protein were down-regulated in U251 cells after transfection with miRNA-99b mimics. However, the abilities of invasion of U251 cells after transfection with miRNA-99b mimics were inhibited. The relative protein expression levels of mTOR in mTOR PsiCHECK group were significantly different from those in negative control group, but the relative expression levels of miRNA-99b had no signifi-cant difference compared with those in negative control group. Over-expression of mTOR restored the abilities of cell invasion in U251 cells, which was reduced by miRNA-99b. The Kaplan—Meier analysis and Log-Rank Test showed that there were significant differ-ences in overall survival (OS) between the miRNA-99b high-expression and low-expression group. Con-clusions The expression level of miRNA-99b is low in human glioma tissue. miRNA-99b may inhibit the cell invasion activity of glioma cell line U251 in vitro via inhibiting mTOR expression.

Objective: To investigate the effects of traditional Chinese medicine, Danzhi decoction, on the expression angiogenesis factors in human endometrial cells during the sequelae of pelvic inflammatory disease (SPID) and explore the role of Danzhi decction in improving the blood stasis microenvironment of SPID. Methods: A three-dimensional (3D) co-culture system including human vascular endothelial cells (VECs), endometrial stromal cells and glandular epithelial cells was established in vitro and treated with Danzhi decoction, sterilized water and aspirin respectively. A Milliplex multifunctional liquid chip technique was used to measure the expression levels of vascular endothelial growth factor (VEGF)-A/C/D, fibroblast growth factor -1/2, angiopoietin-2, epidermal growth factor (EGF), HB-EGF, bone morphogenetic protein-9, endoglin, endothelin-1, granulocyte colony stimulating factor, hepatocyte growth factor, interleukin-8, follistatin, placenta growth factor and leptin. The location of angiogenesis factors was monitored by immunofluorescence labeling and confocal laser scanning microscope 3D reconstruction. Results: Endometrial stromal cells and glandular epithelial cells were isolated and primary cultured for establishing a 3D co-culture system. The levels of VEGF-A/C/D in Danzhi decoction group and aspirin group were significantly lower than those in mock group (. P<0.05), while there was no significant difference between Danzhi decoction group and aspirin group (. P>0.05). Furthermore, the alterative location of VEGF-A/C/D was observed in the cytoplasm of endometrial glandular epithelial cells. Conclusions: Danzhi decoction may inhibit the expression of VEGF in the blood stasis microenvironment of SPID by targeting the cytoplasm of endometrial glandular epithelial cell.

Adolescent ; Adult ; Aged ; Female ; Humans ; Male ; Meningioma ; diagnosis ; Middle Aged ; Retrospective Studies ; Young Adult

Objective To investigate whether gastroesophageal reflux can cause asthma-like pathophysiological changes and its mechanism.Methods Sixty BALB/c mice were randomly divided into 4 groups:group A was gastroesophageal reflux control group,group B was asthma control group,group C was gastroesophageal reflux group,and group D was asthmatic group.The asthmatic models were replicated with ovalbumin(OVA) and aluminum hydroxide,and gastroesophageal reflux models were replicated with hydrochloric acid solution pepsin.After the last inhaling ovalbumin and slow perfusion,the airway pressure was detected,and eosinophil (EOS) and neutrophil granulocyte in bronchial lavage fluid were counted.The flow cytometer was used to determinate IL-4,IFN-γ,and Th1/Th2 ratio changes of spleen cells;Lung tissue and esophagus sections were stained with HE,and pathological changes of lung tissue and esophagus were observed.Results In group C and group D,the airway hyper-responsiveness was significantly increased compared with group A and group B,and the differences were statistically significant (all P ＜ 0.05).In group C and group D,IL-4were significantly increased,while IFN-γand Thl/Th2 ratio were significantly decreased than that in the group A and B group,the differences were statistically significant (all P ＜ 0.05).EOS of group C and group D accounted for a high percentage of total cells and they were significantly higher than that in group A and group B,and the differences were statistically significant (all P ＜ 0.05).Through lung tissue biopsy,in group C and group D,bronchial lumen deformation,infiltration of inflammatory cells around the wall,basement membrane thickening,inflammatory cell infiltration around the wall,peripheral vascular edema,enlarging alveolar cavity,alveolar wall thinning,fracture,part of alveolar fusion into bullace of lung,could be observed.The lung pathological section showed that the endothelial cells in group A and group B were integrated and had no denaturation or necrosis,and there was no inflammation cell infiltrate around.EOS biopsy could be observed in group A and group B of lower esophagus markedly submucosal edema,submucosal inflammatory cell infiltration,and keratin hyper function,visible bacteria group A group B and group D were basic ally normal,with no pathological changes.Conclusions Gastroesophageal reflux can induce Th1/Th2 decreasing,airway hyper-responsiveness and pathophysiological changes similar to asthma.

Objective To supply microanatomic data for the application of the transcallosal-transchoroidal approach, and discuss its clinical value of this application. Methods Twelve adult cadaveric wet brains were carefully dissected under microscope with a magnification ranging for 4-10 times, and the anatomic features of the paracele, the choroids plexus, the choroidal fissure and the diatela were successively explored. Three adult cadaveric fresh heads were studied,simulating the procedures of the transcallosal-transchoroidal approach. Observation was performed on related anatomic structures, and measurement of relative parameter was taken, too. Results In the body of the lateral ventricle, the choroid plexus is located in the medial part of the floor. The choroidal fissure is a C-shaped natural cleft between the thalamus and the fomix, being white translucent membranes, and it could be identified by following the choroid plexus in the lateral ventricle. After carefully opening the taenia fornicis, the contents of the velum interpositum are exposed. The cavity of the third vertricle is reached through opening the roof of the third vertricle between the 2 internal cerebral veins. The distances between the coronal suture at the medial border of the brain and the callosal sulcus were (39.12±3.94) mm; the distances between the coronal suture at the medial border of the brain and the interventricular foramen were (61.53±4.02) mm; the distances between the posterior border of the interventricular foremen and the coalesced area of the fornix and the callosum were (28.65±2.23) mm; the AC-PC distances were (25.94±2.16) mm; the distances between the interventricular foremen and the corpus albicans were (19.62±1.79) mm. Conclusion As compared with other approaches to the third ventricle, the transcallosal-transchoroidal approach, which follows a natural route, is of advantages in few injury and wide exposure, we suggest, therefore, that the transcallosal-transchoroidal approach be extensively adopted in clinical neurosurgery.

OBJECTIVETo observe the effects of neural stem cells (NSCs) transplantation on the brain derived neurotrophic factor (BDNF) after the spinal cord injury (SCI) of rats, and to investigate the mechanism of repairing the SCI by NSCs transplantation.

METHODSNeural stem cells were cultured from the hippocampus of rats' embryo and identified by immunocytochemistry. Seven days after the operation of SCI, the NSCs were transplanted into the injured site. Sixty adult Wistar rats were randomly divided into three groups: SCI cured with NSCs transplantation (group A), SCI received DMEM solution (group B), control group (group C). Then the expression of BDNF of the lesion and neighbor areas were examined by reverse transcsription polymerase chain reaction (RT-PCR) and immunohistochemistry, so as to investigated the mechanism of repairing the SCI after NSCS transplantation.

RESULTSAccording the RT-PCR results analysis, the expression of BDNF mRNA of group A enhanced higher than that of group B on the 1st, 3rd, 5th day after transplantation of NSCs. According the immunohistochemistry results analysis, the expression of BDNF mRNA of group A enhanced higher than that of group B on the 7th, 14th, 28th day similarly.

CONCLUSIONThe transplantation of NSCs can change the tiny-entironment by upregulating the expression of BDNF. It maybe one of the mechanism of repairing the SCI by NSCs transplantation.

Animals ; Brain-Derived Neurotrophic Factor ; genetics ; metabolism ; Cells, Cultured ; Disease Models, Animal ; Gene Expression ; Humans ; Male ; Mesenchymal Stem Cell Transplantation ; Neurons ; metabolism ; transplantation ; Random Allocation ; Rats ; Rats, Wistar ; Spinal Cord Injuries ; genetics ; metabolism ; surgery ; therapy ; Up-Regulation

Objective To explore the microanatomic features of the transcallosal-interforniceal approach, and discuss the value of its clinical application. Methods Fifteen adult cadaveric head specimens were dissected by microsurgical anatomic skills to simulate the procedures of the transcallosal-interforniceal approach. Observation and measurement were performed on related anatomic structures. In clinical, 21 patients with the third ventricle tumors underwent tumor resection via the transcallosal-interforniceal approach. Results Using two points on the cortical surface as references that were located 5 and 7 cm anterior to the central sulcus respectively, mean values of related measurements on P5-foramen of Monro (FM) and P7-FM were obtained as follows: (1) the depth of the interhemispheric fissure was 38.46 and 37.62 ram; (2) the height of the corpus callosum was 7.18 and 7.78 nun; (3) the height of the septum pellucidum was 7.53 and 9.88 mm; (4) the thickness of the fornix was 4.72 and 5.16 mm. Under the operative microscope, the tumors were totally removed in 11 cases,subtotally in 8 cases, and partially in 2 cases. Conclusions The corridor of the transcallosal-interforniceal approach should be limited between the lines of P5-FM and P7-FM. The quantitative information obtained in this study permits the preservation of important anatomic structures in operation, such as the motor strip, genu of the corpus callosum, fornical commissure and anterior commissure. This approach is deserved to be applied generally for providing a quite large operative field,making total tumor removal easier, and decreasing the incidence of postoperative complications.

Adenoma ; blood ; pathology ; Adult ; Estrogens ; blood ; Female ; Human Growth Hormone ; blood ; Humans ; Magnetic Resonance Imaging ; Middle Aged ; Pituitary Neoplasms ; blood ; pathology ; Prolactin ; blood ; Remission, Spontaneous

OBJECTIVETo evaluate the effects and mechanism of radiation-sterilized allogeneic bone sheets in inducing vertebral plate regeneration after laminectomy in sheep.

METHODSTwelve adult male sheep (aged 1.5 years and weighing 27 kg on average) provided by China Institute for Radiation Protection underwent L3-4 and L4-5 laminectomy. Then they were randomly divided into two groups: Group A (n=6) and Group B (n=6). The operated sites of L4-5 in Group A and L3-4 in Group B were covered by "H-shaped" freeze-drying and radiation-sterilized allogeneic bone sheets (the experimental segments), while the operated sites of L3-4 in Group A and L4-5 in Group B were uncovered as the self controls (the control segments). The regeneration process of the vertebral plate and the adhesion degree of the dura were observed at 4, 8, 12, 16, 20 and 24 weeks after operation. X-ray and CT scan were performed in both segments of L3-4 and L4-5 at 4 and 24 weeks after operation.

RESULTSIn the experimental segments, the bone sheets were located in the anatomical site of vertebral plate, and no lumbar spinal stenosis or compression of the dura was observed. The bone sheets were absorbed gradually and fused well with the regenerated vertebral plate. While in the control segments, the regeneration of vertebral plate was not completed yet, the scar was inserted into the spinal canal, compressing the dura and the spinal cord, and the epidural area almost disappeared. Compared with the control segments, the dura adhesion degree in the experimental regenerated segments was much milder (P less than 0.01), the internal volume of the vertebral canal had no obvious change and the shape of the dura sack remained well without obvious compression.

CONCLUSIONSFreeze-drying and radiation-sterilized allogeneic bone sheets are ideal materials for extradural laminoplasty due to their good biocompatibility, biomechanical characteristics and osteogenic ability. They can effectively reduce formation of post-laminectomy scars, prevent recurrence of post-laminectomy spinal stenosis, and induce regeneration of vertebral plates.

Animals ; Bone Transplantation ; methods ; Laminectomy ; methods ; Regeneration ; Sheep ; Spinal Stenosis ; prevention & control ; Spine ; physiology ; Transplantation, Homologous

Objective To understand epidemiological characteristics of chronic obstructive pulmonary disease(COPD)in people aged over 40 years in a rural area of Tianjin.Methods Using cluster sampling,1 508 subjects over 40 years old at five villages in Xinkaikou Township,Baodi District,Tianjin were investigated with respiratory questionnaire,lung function test and physical examination.Confirmed patients with COPD were examined by chest roentgenography and electrocardiography.Results One hundred and forty-two subjects in that area suffered from COPD,with prevalence of 9.4%,24 of them (16.9%)were diagnosed as cor pulmonale.Prevalence of COPD increased with age,higher in men (13.5%)than that in women(6.2%),higher in smokers(12.2%)than that in non-smokers(7.2%), higher in those with family history(21.4%)than that in those without it(8.45%),and higher in those with coughing history during their childhood(75.0%)than that in those without it(9.2%),all with a P-value of less than 0.01.Univariate analysis showed that out-door air pollution,cooking,time length of burning firewood during cooking,smoking,coughing history during childhood,gender,age,family history all were predisposing factors for COPD.Multivariate analysis with logistic regression model showed that gender,age, family history were independently predisposing factors for COPD.Quality of life was better in non-COPD subjects than in those with COPD,with statistically significant difference.Conclusions Prevalence of COPD was relatively higher in people of rural Tianjin,with gender,age,family history and outdoor air pollution as main risk factors.