Objective To investigate the possibility of ectomesenchymal stem cell of human embryo facial process in differentiating into osteoblasts.Methods Ectomesenchymal stem cells of human embryo facial process were isolated and cultured in mineralized promoting solution containing 10 mmol/L β-glycerophosphate, 100 μg/ml ascorbic acid and 10 nmol/L dexamethasone supplemented with 15% FBS. The morphological change was observed by phase contrast microscopy. The characteristics of cells was identified by immunohistochemistry assay. Alkaline phosphatase activity was tested and the form of mineralized nodules was tested with Von Kossa staining. The expression of osteocalcin was identified by RT-PCR.Results There were significant changes in the shape of the cells after 3 days cultured in mineralized promoting solution. The cells became larger and the shape changed from fibroblast-like to multilateral. The result for anticollogen typeⅠstaining was positive. The alkaline phosphatase activity increased. Mineralized nodules were formed aftercultured 25 days by Von Kossa staining. RT-PCR assay showed induced cells expressed osteocalcin.Conclusion Ectomesenchymal stem cells of humanembryo facial process can be induced to differentiate into osteoblasts by mineralized promoting solution.
Objective To investigate the effect of NGF on fracture heal ing, and to study the role of BMP-2 induced osteoblast. Methods Sixty cleaned male Kunming mice (aging 6-8 weeks and weighing 23-25 g) were made fracture models in the middle of femoral shaft and randomly divided into four groups (groups A, B, C and D, n=15). Fracture was treated with NGF/ normal sal ine, BMP-2, BMP-2 /NGF/normal sal ine, and normal sal ine in groups A, B, C and D, respectively. After 14, 21 and 28 days, the specimens were selected from 5 mice each group to do the biochemical and histological analysis. Beforethe mice were killed, the arteriovenous blood was taken from their eye-ball to test the ALP activity. Results After 14 days,21 days and 28 days, the gross observation showed that the size and hardness of bone tissue, and callus tissue growth increased in groups A, B and C order and were higher than those in group D; the X-ray films showed that the calcified area increased in groups A, B and C order and were higher than those in group D; the histological observation showed that the trabecular maturity increased in groups A, B and C order and were higher than those in group D. The osteoblast area, the gray degree value of the radiographs in callus tissue, the ALP contents of serum and callus tissue, calcium content of callus tissue and net weight of callus were higher in groups A, B and C than in group D. There were significant differences (P lt; 0.05) in osteoblast area and gray degree values of the radiographs at 14, 21 and 28 days; in ALP contents of serum at 14 days; in ALP contents of callus tissue at 14 days and 21 days; in calcium content of callus tissue at 21 days and 28 days among 4 groups. There were significant differences in net weight of callus between groups B, C and groups A, D at 14 days (P lt; 0.05). At 21 days and 28 days, the trabecular surface index of osteoblast, the average trabecular volume and the mean trabecular width decreased as time went on, having an increase order of groups A, B, C and was higher in groups A, B, C than in group D, showing significant differences among 4 groups (P lt; 0.05). Conclusion NGF promotes the heal ing of fractures. NGF possesses synergistic effect on ectopic bone formation induced by BMP-2.
ObjectiveTo study the possibility of the C17.2 neural stem cells (NSCs) differentiating into neural cells induced by serum-free condition medium of olfactory ensheathing cells (OECs) and to detect the cell viability of the differentiated cells. MethodsOECs were isloated and cultured from the olfactory bulbs of 3-day-old postnatal mouse to prepare serum-free condition medium of OECs. After C17.2 NSCs were cultured with H-DMEM/F12 medium containing 15% FBS and the cell fusion reached 80%, the 3rd passage cells were induced by serum-free condition medium of OECs in the experimental group, by H-DMEM/F12 in the control group, and non-induced C17.2 NSCs served as the blank control group. The growth condition of cells was observed with inverted microscope. After 5 days, the immunofluorescence staining[microtubule-associated protein 2 (MAP-2) and β-tubulin-Ⅲ] and Western blot (Nestin, β-tubulin-Ⅲ, and MAP-2) were carried out to identify the neural cells derived from NSCs. The cell viabilities were measured by MTT assay and the quantity of lactate dehydrogenase (LDH) release in the medium. ResultsIn the experimental group, the C17.2 NSCs bodies began to contract at 24 hours after induction, and the differentiated cells increased obviously with long synapse at 3 days after induction; in the control group, the cell morphology showed no obvious change at 24 hours, cell body shrinkage, condensation of nuclear chromatin, and lysis were observed at 3 days. The immunofluorescence staining showed that β-tubulin-Ⅲ and MAP-2 of C17.2 NSCs were positive at 5 days after induction, and Western blot suggested that the expression of Nestin protein declined significantly and the expressions of β-tubulin-Ⅲ and MAP-2 protein were increased in the experimental group, showing significant differences when compared with those in the control group and blank control group (P<0.05). The LDH release and the cell viability were 130.60%±6.86% and 62.20%±3.82% in the experimental group, and were 178.20%±5.44% and 18.00%±3.83% in the control group respectively, showing significant differences between 2 groups (P<0.05). The LDH release and the cell viability of experimental group and control group were significantly lower than those of blank control group (100%) (P<0.05). ConclusionNeurotrophic factors from OECs play an important role in inducing C17.2 NSCs differentiation into neural cells and keeping the viability of differentiated cells after induction.
Increasing evidences show that a gradual trend away from deep hypothermia toward moderate hypothermic circulatory arrest, which has been proved to be safe and effective in clinic. By summarizing and analyzing the research progress and applying status of the moderate hypothermia circulatory arrest with selective antegrade cerebral perfusion, the article aims at promoting the application of this tenique as a cerebral protection strategy in aortic arch surgery for adults in China.
Objective To investigate the clinical characteristics of neutrophilic asthma ( NA) .Methods NA patients were collected from the out-patient and in-patient departments of Respiratory Diseases of Xinqiao Hospital between January 2010 and December 2010. The results of the medical records,pulmonary function tests, and induced sputum cytology were analyzed retrospectively. Results The NA patients with neutrophil percent ≥ 61% accounted for 33. 1% ( 51 /154 ) of all the asthmatics patients detected by induced sputumin the same period, and 45 cases with complete records were included. Of the 45 cases recruited, 20 cases ( 44. 4% ) were in-patients,2 cases ( 4. 4% ) were with controlled asthma, 3 cases ( 6. 7% ) were with cough variant asthma, 30 cases ( 66. 7% ) were female, 12 cases ( 26. 7% ) were atopic patients, and 27 cases ( 60% ) had acute exacerbation. The age of onset of 35 patients ( 77. 8% ) were after 12 years. FEV1% pred lt; 60% and gt; 80% was obtained in 55. 9% ( 19/34) and 14. 7% ( 5 /34) of patients respectively. The result of bronchodilator test was positive in 64% ( 16/25) of patients, and mean increase in FEV1 was 11. 7% . The percentage of neutrophil and eosinophil was ( 74. 5 ±9. 1) % and ( 2. 4 ±2. 5) % respectively in induced sputum, and 35. 6% ( 16/45) of the patients had increased eosinophil percentage ( gt;3% ) . Conclusions In our study, most of NA is severe and acute exacerbation asthma, and its clinical features are various. The mechanismand clinical significance of increased neutrophils in asthmatic patients are unclear and more studies are needed.
Objective To study the relationship between the expression ratio of induced nitric oxide synthase (iNOS) over glial fibrillary acidic protein (GFAP) and the time of injury after brain concussion in rat, in order to acquire a new visual angle for determining injury time of cerebral concussion. Methods Eighty-five healthy Sprague-Dawley rats were divided into three groups randomly: model group (n=25), experimental group (n=55), and control group (n=5). The rats in the model group were used to confirm the attack hight to make the model of brain concussion; according to the time of execution, rats in the experimental group were then subdivided into 11 groups with 5 rats in each subgroup, and their execution time was respectively hour 0.5, 1, 3, 6, 12, 24, 48, 96, 168, 240, and 336; the rats in the control group were executed after fed for 24 hours. After the model of cerebral concussion was established through freefalling dart method, hematoxylin-eosin staining and immunohistochemistry staining of iNOS and GFAP were conducted for the brain of the rats. All related experimental results were studied by using microscope with image analytical system and homologous statistics. Results The ratio of positive expression of iNOS over that of GFAP increased gradually during hour 0.5- 3 after injury in brain (from 5.03 to 10.47). At the same time, the positive expression of iNOS increased significantly (from 14.61% to 37.45%). However, the increase of the positive expression of GFAP was not obvious. Between hour 3 and 12, the ratio began to decline to 4.98, which was still at a high level, and during the same time period, the positive expressions of iNOS and GFAP also experienced the same change pattern. Later, the ratio began to decline between hour 12 and 336 after injury (from 4.98 to 0.95). All ratios at this time were lower than those between hour 0.5 and 12. The positive expression of iNOS and GFAP both increased to a climax before declining. Conclusions The ratio of positive expression of iNOS over GFAP and the respective change pattern of iNOS and GFAP can be used as the evidence of estimating the injury time of cerebral concussion. We can use the ratio of two or more markers to provide a new visual angle for concluding the concussion injury time.
ObjectiveTo explore the characteristics of induced sputum microbiome in the patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD).MethodsInduced sputum samples from 55 patients with AECOPD and 45 patients with stable COPD were analyzed by sequencing of 16S rRNA gene. Microbiota was measured by alpha diversity, beta diversity and LDA effect size analysis (LefSe).ResultsThe microbiome diversity of induced sputum in the AECOPD group was lower than that in the stable COPD group. The microbiome richness in the AECOPD group was higher than that in the stable COPD group. The microbiome structure changed in the AECOPD group compared with the stable COPD group. The proportion of some common pathogens got enriched. The levels of hypersensitive C reactive protein (hs-CRP), interleukin-8 (IL-8), tumor necrosis factor alpha (TNF-α) and Global Initative for Chronic Obstructive Lung Disease (GOLD) grade were negatively related to the diversity of microbiome in the AECOPD group.ConclusionsThe microbiome diversity of induced sputum in AECOPD patients is decreased, and is negatively correlated with the levels of hs-CRP, IL-8, TNF-α and GOLD grade. There are differences in the microbiome structure between AECOPD and stable COPD patients. Some enrichment of common pathogens are found in the induced sputum of patients with AECOPD. These results suggest that there is a significant bacterial dysbiosis in patients with AECOPD.
Objective To explore effect of platelet-rich plasma (PRP) on rabbit BMSCs differentiation into SC in vitro and to detect secretory function of the differentiated cells. Methods BMSCs isolated from 5 mL bone marrow of 2-montholdNew Zealand white rabbit were cultured using density gradient centrifugation and adherence screening methods. A total of 5 mL femoral vein blood was obtained from rabbits to prepare PRP using modified Appel method. The BMSCs at passage 3 were divided into three groups: the combined induction group, in which the cells were cultured with complete medium containing PRP after β-mercaptoethanol and retinoic acid inductions; the simple induction group, in which the cells were cultured with L-DMEM complete medium without PRP afterβ-mercaptoethanol and retinoic acid induction; the control group, in which the cells were cultured with L-DMEM complete medium. Growth condition of the cells in each group was observed using inverted microscope. cell identification was conducted at 4, 7, 9, and 11 days after culture using immunofluorescence staining method, and NGF content was detected by ELISA method. NGF mRNA expression was assayed by RT-PCR 11 days after culture. Results Most cells in the combined induction and the simple induction group were out of BMSCs typical cell morphology 4 days after culture; cells in the combined induction group were out of BMSCs typical cell morphology and changed into cells resembl ing SC in terms of morphology and contour 9 days after culture. The cells in the control group showed no obvious morphological changes. S-100 protein expression in the cells was evident in the combined induction and the simple induction group at each time point after induced culture; the positive expression rate of cell in each group was increased over time, and significant differences were evident between the combined induction group and the simple induction group 7, 9, and 11 days after culture (P lt; 0.05). Control groupwas negative for the expression. There were significant differences when comparing the control group with the combined induction group or the simple induction group in terms of NGF content at each time point (P lt; 0.01). Significant difference was evident between the combined induction group and the simple induction group 7, 9, and 11 days after culture (P lt; 0.05), and no significant difference was noted 4 days after culture (P gt; 0.05). Relative intensity of NGF mRNA expression in the combined induction group was greater than that of the simple induction group 11 days after culture (P lt; 0.05). Conclusion Rabbit BMSCs can differentiate into SC excreting NGF under certain induction condition in vitro. PRP can remarkably promote BMSCs differentiation into SC.
In transcranial magnetic stimulation (TMS), the conductivity of brain tissue is obtained by using diffusion tensor imaging (DTI) data processing. However, the specific impact of different processing methods on the induced electric field in the tissue has not been thoroughly studied. In this paper, we first used magnetic resonance image (MRI) data to create a three-dimensional head model, and then estimated the conductivity of gray matter (GM) and white matter (WM) using four conductivity models, namely scalar (SC), direct mapping (DM), volume normalization (VN) and average conductivity (MC), respectively. Isotropic empirical conductivity values were used for the conductivity of other tissues such as the scalp, skull, and cerebrospinal fluid (CSF), and then the TMS simulations were performed when the coil was parallel and perpendicular to the gyrus of the target. When the coil was perpendicular to the gyrus where the target was located, it was easy to get the maximum electric field in the head model. The maximum electric field in the DM model was 45.66% higher than that in the SC model. The results showed that the conductivity component along the electric field direction of which conductivity model was smaller in TMS, the induced electric field in the corresponding domain corresponding to the conductivity model was larger. This study has guiding significance for TMS precise stimulation.
OBJECTIVE To testify the inductive osteogenesis of allogeneic bone matrix gelatin (BMG) in promoting intervertebral fusion. METHODS The gelatin sponge, allogeneic BMG, decalcified bone matrix (DBM) and alcohol conserved bone were implanted respectively into the intervertebral space of rabbit, whose intervertebral discs were removed before implantation. The intervertebral spaces were evaluated by X-ray and histological examination at 4, 8, and 12 weeks after operation. RESULTS No obvious immune rejection was observed. Amounts of new bone were formed in the intervertebral spaces at 4 and 8 weeks. And complete infusion of the intervertebral spaces were appeared at 12 weeks. CONCLUSION Allogeneic BMG can promote bone fusion of intervertebral spaces through osteoinduction, which suggests that allogeneic BMP is an ideal substitute for bone replacement.