OBJECTIVETo observe the three-dimensional distribution of vessels, and to establish a new method for measurement of blood flow velocity in mice cerebral cortex using two-photon laser scanning microscopy and fluorescence probe labeling technique.

METHODSThe mouse was made cranial window surgery and injected Texas-Red through tail vein after anesthetized. The three-dimensional imaging of vessel was obtained through z-stack scanning, and blood flow velocity was quantified through line scanning.

RESULTSWe could detect vascular distribution for more than 500 µm depth using two-photon microscopy. The velocity of blood flow was (0.59 ± 0.12) mm/s in capillary.

CONCLUSIONThe method for observing the brain blood flow by two-photon microscopy was established, which could achieve quantification of single vascular blood flow velocity and provide experimental evidence for basic research and medical applications.

Animals ; Blood Flow Velocity ; Brain ; blood supply ; Capillaries ; Cerebrovascular Circulation ; Fluorescent Dyes ; Hemodynamics ; Mice ; Microscopy, Fluorescence

Treatment based on syndrome differentiation is an essential feature of traditional Chinese medical diagnosis. The interventions based on changes of syndrome types in randomized controlled trials are complicated, leading to the difficulty of blind method enforcement. This article described a double-blind method. It could be used in randomized controlled trials under the condition of different syndrome types and different medications. It numbered drugs in two stages, and in two phases to achieve double-blind. This method not only guaranteed investigators and subjects to be in blinded conditions, but also achieved using different medications for patients of different syndromes. It also caused no drug waste. It was scientific and feasible.
Double-Blind Method ; Humans ; Medicine, Chinese Traditional ; Randomized Controlled Trials as Topic ; Single-Blind Method

Objective To study the scanning technique of 16-detector multic-slice spiral CT (MSCT) for combined pulmonary artery and deep vein of lower limb in pulmonary thromboembolism (PE) patients.Methods Forty suspected pulmonary thromboembolism patients were performed both pulmonary artery angiography (CTA) and indirect deep vein venography (CTV) on 16-detector MSCT. The parameters of the latter as following :total contrast volume 120-150 ml, injection rate 4.0-4.5 ml/s (from antecubital vein), delay time 4.0 for CTA 20-23 s, CTV 120-180 s, collimation for CTA 1.25 mm and 0.625 mm, CTV 2.5 mm, scan range of CTV: from popliteal vein to the level of bilateral renal vein into the inferior vena cava. Postprocessing include MPR, MIP, and VR. The test was used to analyzed the images.Results Twenty five patients had both pulmonary thromboembolism(PE) and deep vein thromboembolism (DVT), 8 patients had only DVT, 2 had only PE, and 5 had neither. There was no difference between different collimation in depicting thrombus. The CT value number of enhanced pulmonary artery and lower deep vein was obviously higher than the thrombus. The value of MPR, MIP, VR for PE was 100%, 100%, and 65%, The value of MPR,MIP,VR for DVT is 100%, 60%, and 50%.Conclusion The technique of combined pulmonary CTA and deep vein CTV of 16-detector MSCT will provide a new modality for pulmonary thromboembolism patients.

Diagnostic whole body scan (Dx-WBS) with 131I and serum Tg level are the main parameters to evaluate the effectiveness of thyroid remnant ablation in patients with DTC.Undetectable Tg and positive radioiodine uptake in the thyroid bed (Tg-/Dx-WBS+) may be found in some patients.However,the clinical significance is uncertain.A small amount of thyroidal remnant,a small DTC lesion,increased expression of NIS gene and autoimmune inflammation may all result in Tg-/Dx-WBS+.A wait-and-watch approach without rushing for high-dose radioiodine treatment might be a more reasonable approach for these patients.

Objective To evaluate the applications of magnetic resonance cholangiopancreatography (MRCP) after fat meal in the preoperative evaluation of biliary anatomy of living liver donors.Methods Fifty cases of the preoperative donors for living liver transplantation were included and all had the corresponding intraoperative cholangiography (IOC) information. The MRCP of the donors for living liver transplantation was performed before and after fat meal (two fried eggs). The visualization and diameter of the secondary bile duct were analyzed before and after the fat meal. The results of the biliary branching pattern by MRCP after fat meal were compared with the corresponding IOC results. The accuracy, sensitivity,specificity, positive predictive value and negative predictive value of MRCP after the fat meal in distinguishing normal and any type of variant biliary anatomy were calculated. Results In all cases,82% of the 50 cases in MRCP before the fat meal could meet the diagnosis needs of the preoperative evaluation,and 100% of the 50 cases in MRCP after the fat meal could meet the diagnosis needs. There was significant difference in the demonstration quality and diameter of the secondary bile duct in MRCP before and after the fat meal (P＜0. 05). MRCP showed accurate anatomy of the biliary system, using IOC as the reference standard, in 49(98%) subjects. The sensitivity, specificity, positive predictive value and negative predictive value of MRC in distinguishing normal and any type of variant biliary anatomy were 98%,94. 7%, 100%, 10% and 96. 9%,respectively. Conclusion The MRCP after fat meal can clearly demonstrate the secondary bile duct and perfectly meet the needs of the preoperative evaluation of the living liver transplantation. The MRCP after fat meal and routine MRCP should be considered complementary to one another in order to avoid complications in living liver transplantation donors.

19 cases with vertical alveolar bone resoption wer examined by CBCT and treated by orthodentic therapy, the changes of alveolar ridge height, bone defect and bone mineral density were analysed. After treatment, the defect area of alveolar bone was decreased(P＜ 0. 05), the density of defect area of alveolar bone was increased(P＜ 0. 05). CBCT plays an important role in the evaluation of vertical alveolar bone absorption and reconstruction treated by orthodontic therapy.

Population pharmacokinetics of vancomycin (VAN) in the Chinese patients was described by using nonlinear mixed-effects modeling (NONMEM). 619 VAN serum concentrations data from 260 patients including 177 males and 83 females were collected separately from two centers. A one-compartment model was used to describe this sparse data. No significant difference was observed between two center datasets by introducing SID covariate. The final model was as CL= (θ (base0+ θ(max) x(1 -e(-θ(Age)(Age/72) and V = θ x θ (Age)(Age/72). The creatinine clearance (CL(Cr)) and Age were identified as the most significant covariate in the final model. Typical values of clearance (CL) and volume of distribution (V) in the final model were 2.91 L x h(-1) and 54.76 L, respectively. Internal model validation by Bootstrap and NPDE were performed to evaluate the robustness and prediction of the final model. The median and 95% confidence intervals for the final model parameters were based on 1000 Bootstraps. External model evaluation was conducted using an independent dataset that consisted of 34 patients to predict model performance. Pharmacodynamic assessment for VAN by AUC (0-24 h) to MIC ratios of over 400 was considered to be the best to predict treatment outcomes for patients. AUC (0-24 h) was calculated by clearance based on the above population model. The results indicate that the conventional dosing regimen probably being suboptimal concentrations in aged patients. The approach via population pharmacokinetic of VAN combined with the relationship of MIC, Age, CL(Cr) and AUC(0-24 h)/MIC can predict the rational dose for attaining efficacy.
Aged ; Asian Continental Ancestry Group ; Female ; Humans ; Male ; Models, Biological ; Nonlinear Dynamics ; Vancomycin ; pharmacokinetics

Objective To investigate CTGF changes in the expression of the gum tissue before and after orthodontic treatment, and to preliminarily explore the modification mechanism of gingival tissue and the effectiveness of the intervention measures. Methods 72 male Sprague-Dawley rats of 12-week-old, weight about 250 g-300 g, randomly divided into 6 groups: blank control group (A), healthy teeth orthodontic group (B), low functional group (C), low occlusal function teeth orthodontic group (D), combined intervention group (E), bite orthodontic intervention group (F). The results of the study were compared at 1 w, 2 w, 4 w, 6 w.Results (1) HE staining results showed the atrophy of the gingival tissue, which suggested that occlusal hypofunction SD rats model were successfully established. (2) Fluorescence quantitative results of CTGF in gingival tissue: 6 w: group B was higher than group D and group F (P＜0.05). Conclusion (1) the expression of CTGF in the low occlusion group was lower than that of the normal control group with time, and the gum tissue was vulnerable to atrophy. (2) It remains to be further studied whether the bite force recovery is effective.

OBJECTIVETo observe the mitochondrial fragmentation and the expression of mito-fusion 1 gene in the cortical neurons of rats with chronic fluorosis, and to reveal their roles in mitochondria damage to neurons due to chronic fluorosis.

METHODSSD rats were divided randomly into three groups of 20 each (a half females and a half males housed individually in stainless-steel cages), and fed with the different doses of fluoride containing in drinking water (untreated control containing 0 mg/L fluoride, and low-fluoride and high supplemented with 10 and 50 mg/L fluoride, respectively). After 3 or 6 months exposure, the mitochondrial morphology of the neurons in rat brains were observed by transmission electron microscopy (TEM), then the expression of mitochondrial fusion gene, Mfn1, were detected by immunohistochemistry and western-blotting, respectively.

RESULTSDental fluorosis was obvious in the rats exposed to excessive fluoride in their drinking water, that is, (16 rats out of 20) numbers of I° detal fluorosis in the low-fluoride group, and (11 rats out of 20) numbers of I° and (9 rats out of 20) numbers of II° detal fluorosis in the high-fluoride group were observed after 3 months exposure. Moreover, (14 rats out of 20) numbers of I° and (6 rats out of 20) numbers of II° detal fluorosis in the low-fluoride group and (6 rats out of 20) numbers of Io, (13 rats out of 20) numbers of II°, and (1 rats out of 20) numbers of III° detal fluorosis in the high-fluoride group were observed after 6 months exposure. And both of untreated controls without detal fluorosis were also observed. The urinary level of fluoride in the low-fluoride group (3.30 ± 1.18) mg/L and in the high-fluoride group (5.10 ± 0.35) were observed after 3 months exposure (F = 3.18, P < 0.05). Moreover, the urinary level of fluoride in the low-fluoride group (4.16 ± 1.39) mg/L and in the high-fluoride group (5.70 ± 1.70) mg/L were also observed after 6 months exposure (F = 3.17, P < 0.05). The normal mitochondrial morphology of neurons in rats without fluorosis was observed after 3 and 6 months, while the abnormal mitochondrial morphology of neurons with fluorosis was shown, presenting mitochondrial fragmentation with swollen cristae and even the fragmented, shortened or stacked punctuate membranes (section observation of three bullous mitochondrial-mitochondrial fission process) by TEM. As compared with controls (53.0 ± 4.54 and 1.21 ± 0.18) at the experiment period of 3 months, Mif1 protein analysis with immunocytochemical (the numbers of positive cells: 51.09 ± 6.25) and western-blotting (1.22 ± 0.26) were no significant difference for low fluoride group (t = 1.7, 1.1, P > 0.05); Mif1 protein analysis with immunocytochemical (the numbers of positive cells: 59.71 ± 5.64) and western-blotting (1.66 ± 0.20) were significantly increasing for high fluoride group (t = 2.1, 2.1, P < 0.05). As compared with controls (36.43 ± 4.04 and 1.00 ± 0.13) at the experiment period of 6 months, Mif1 protein analysis with immunocytochemical (the numbers of positive cells 20.05 ± 4.55 and 17.10 ± 3.86) and western-blotting (0.64 ± 0.08 and 0.39 ± 0.06) were significantly decreasing for the two fluoride group (t = 2.1, 2.2; 2.2, 2.2 respectively, all P value were < 0.05).

CONCLUSIONSTaking excessive amount of fluoride might result in the mitochondrial fragmentation for the changed expression of Mfn1, and the neurons damage from the chronic fluorosis might be associated with the dysfunction of mitochondrial fusion.

Animals ; Drinking Water ; chemistry ; Female ; Fluoride Poisoning ; metabolism ; pathology ; Fluorosis, Dental ; metabolism ; Male ; Membrane Proteins ; metabolism ; Mitochondria ; pathology ; Mitochondrial Proteins ; metabolism ; Neurons ; metabolism ; pathology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo study mechanisms of reduction of the malignant activities of human naso-pharyngeal carcinoma cell CNE-2L2 induced by ectopic expression of BCSC-1 gene.

METHODSDNA was stained with propidium iodide and assayed upon a flow cytometer. Chromosomes were stained with Hoechest 33258. Adhesion of CNE-2L2 cells was detected by cell aggregation test. Protein expression on CNE-2L2 cells was examined by Western blot.

RESULTSCell cycle analysis showed that the percentage of CNE-2L2 cells was 55.1%, 43.4%, and 39.4% in G0/G1 phase, 25.2%, 28.7%, and 30.9% in S phase, and 19.7%, 27.9%, and 29.7% in G2/M phase for the cell with ectopic expression of BCSC-1 gene, wild type cell (W cells), and the cell transduced with the mock (M cell). Many mitotic cells were found in W cells and M cells. In contrast, almost no mitotic cell was observed in the cells with ectopic expression of BCSC-1 gene. Ectopic BCSC-1 expression resulted in cell aggregation, enhanced expression of E-cadherin, cx-catenin, and p53.

CONCLUSIONSEctopic BCSC-1 expression causes enhancement of adhesion of CNE-2L2 cells associated with enhanced expression of E-cadherin and alpha-catenin, arrest of cell in G1 phase, which may be associated with enhanced expression of p53. These alteration may play a role in the reduction of malignant activities of the cells with ectopic expression of BCSC-1 gene.

Cell Adhesion ; Cell Cycle ; physiology ; Cell Line, Tumor ; Humans ; Nasopharyngeal Neoplasms ; Neoplasm Proteins ; biosynthesis ; genetics