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.
Objective To observe the expression of genes related to hereditary retinal diseases (IRD) in human microglia (hMG). MethodsA experimental study. Efficient differentiation of human induced pluripotent stem cells (iPSC) into hMG. Identification of octamer-binding transcription factor 4 (OCT4), sex-determining transcription factor 2 (SOX2), Nanog homeobox (NANOG), stage-specific embryonic antigen-4 (SSEA4), alpha-fetoprotein (AFP), α-smooth muscle actin (α-SMA) as markers associated with iPSC dryness and pluripotency by immunofluorescence staining Glial fibrillary acidic protein (GFAP); hMG associated marker transmembrane protein 119 (TMEM119), purinergic receptor P2Y12 (P2RY12), and allograft inflammatory factor 1 (IBA1). The proportion of CD11b+ and CD45+ cells was detected by flow cytometry. Mature hMG was collected and stimulated with lipopolysaccharide for 0, 4, 8 and 12 h, and were divided into groups 0 h, 4 h, 8 h and 12 h, respectively. Total RNA samples from the 4 groups were extracted for transcriptome sequencing, and the persistently significant differentially expressed genes (DEG) were screened. Real-time quantitative polymerase chain reaction (qPCR) was used to verify and analyze the expression of DEG mRNA. The two-tailed Student t test was used for comparison between the two groups. ResultsiPSC expressed the dry related markers OCT4, SOX2, NANOG and SSEA4, and differentiated into endoderm, mesoderm and ectoderm, expressing the corresponding markers AFP, α-SMA and GFAP, respectively. iPSC formed embryoid bodies under specific culture conditions, and then differentiated into hMG, and hMG expressed related markers TMEM119, P2RY12 and IBA1 by immunofluorescence staining. The double positive ratio of CD11b+ and CD45+ was > 95%. Transcriptomic analysis showed that the expression of 18 DEG in hMG stimulated by LPS was changed. qPCR test results showed that compared with group 0 h, mRNA expressions of Toll-like receptor 4 (TLR4), phosphoglycerate kinase 1, disintegrin and metallopeptidase domain 9 (ADAM9) in LPS stimulated group 4 h were significantly increased (t=25.43, 15.54, 6.26; P＜0.01). The mRNA expression levels of MER proto-oncogene tyrosine kinase (MERTK), non-hydrolase domain containing lysophospholipase 12 (ABHD12), retinal dehydrogenase 11 (RDH11), DNA damage autophagic regulator 2 (DRAM2) decreased (t=5.94, 14.14, 8.21, 6.97; P＜0.01), and the differences were statistically significant. Compared with group 0 h, mRNA expressions of RDH11, MERTK, ABHD12, DRAM2 and ADAM9 in group 8 h stimulated by LPS were significantly decreased, with statistical significance (t=25.97, 5.47, 43.97, 38.40, 3.84; P＜0.05). Compared with the group 0 h, the mRNA expressions of TLR4, ADAM9, MERTK, ABHD12, RDH11 and DRAM2 in the 12 h stimulated group were significantly decreased, and the differences were statistically significant (t=6.39, 46.11, 5.34, 14.14, 25.97, 25.65; P＜0.05). ConclusionIRD-related genes may be involved in the occurrence and development of IRD by regulating the function of hMG.
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.
ObjectiveTo investigate the differentiation of rat adipose-derived stem cells (ADSCs) into neuronlike cells by indirect co-culture with Schwann cells (SCs) in vitro so as to look for the ideal seed cells for tissue engineering. MethodsSCs were isolated from sciatic nerves of 1-2 days old Sprague-Dawley rats with enzymatic digestion method. Immunofluorescence staining was used to identify SCs with the marker S-100. ADSCs were isolated from the epididymal fat pads of adult male Sprague-Dawley rats by means of differential attachment. And the cell phenotypes (CD29, CD34, CD45, CD73, CD90, and CD105) of ADSCs at passage 3 were determined by flow cytometry analysis. Primary SCs and ADSCs at passage 3 were co-cultured at a ratio of 2:1 in Transwell culture dishes (experimental group), and ADSCs cultured alone served as control group. Immunofluorescence and flow cytometry were adopted to investigate the neural differentiation of ADSCs at 14 days. The expression differences for neuron-specific enolase (NSE), microtubule-associated protein 2 (MAP2), neuronal nuclei protein (NeuN), and glial fibrillary acidic protein (GFAP) were detected, and the percentage of positive cells was calculated. ResultsADSCs were successfully extracted and can passage in a considerable large amount. Flow cytometry analysis showed that ADSCs at passage 3 were positive for CD29, CD90, CD73, and CD105 expression, but negative for CD34 and CD45 expression. The ADSCs of the experimental group showed contraction of nucleus, increasing of soma refraction, and several long and thick protrusions of cell body. The cell shape had no obvious change in the control group. Both immunofluorescence and flow cytometry analysis results showed the expressions of MAP2, NSE, NeuN, and GFAP at 14 days after co-cultured with SCs, and the positive cell ratios were significantly higher than those in the control group (P<0.01). ConclusionCo-culture with SCs not only can promote the survival regeneration of ADSCs, but also can induce the differentiation of ADSCs into neuron-like cells.
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 evaluate the effectiveness of induced membrane technique in the treatment of infectious bone defect. Methods Thirty-six patients (37 bone lesions) with infectious bone defects were treated with induced membrane technique between January 2011 and June 2014. There were 28 males and 8 females with an average age of 36 years (range, 20-68 years). All bone defects were post-traumatic infectious bone defect. The bone defect was located at the tibia and fibula in 24 cases (25 bone lesions), at femurs in 6 cases (6 bone lesions), at ulnas and radii in 2 cases (2 bone lesions), at calcanei in 3 cases (3 bone lesions), and at clavicle in 1 case (1 bone lesion). The average time between onset and the treatment of induced membrane technique was 6.2 months (range, 0.5-36.0 months); 15 patients were acute infections (disease duration was less than 3 months). At the first stage, after the removal of internal fixator (applicable for the patients who had internal fixation), complete debridement of infection necrotic bone tissue and surrounding soft tissue was performed and the bone defects were filled with antibiotic-impregnated cement spacers. If the bone was unstable after debridement, external fixator or plaster could be used for stabilization. Patients received sensitive antibiotics postoperatively. At the second stage (usually 6-8 weeks later), the cement spacer were removed, with preservation of the induced membrane formed by the spacer, and filled the bone defect with autologous iliac bone graft within the membrane. Results The hospitalization time after debridement was 17-30 days (mean, 22.2 days), and the hospitalization time after the second stage was 7-14 days (mean, 10 days). All the flaps healed uneventfully in 16 cases treated with local flap transposition or free flap grafting after debridement. One patient of femur fracture received Ilizarov treatment after recurrence of infection at 11 months after operation; 1 patient of distal femoral fracture received amputation after recurrence of infection at 1 month after operation; 1 patient of distal end of tibia and fibula fractures received ankle arthrodesis after repeated debridements due to the recurrence of infection; 1 patient of tibia and fibula fractures lost to follow-up. The other 32 patients (33 bone lesions) were followed up 1-5 years (mean, 2 years) without infection recurrence, and the infection control rate was 91.7% (33/36). All the patients had bony union, and the healing time was 4-12 months (mean, 7.5 months); no refracture occurred. One patient of femur bone defect had a lateral angulation of 15° and leg discrepancy of 1.5 cm. Superficial pin infection was observed in 7 cases and healed after intensive wound care and oral antibiotics. Adjacent joint function restriction were observed in 6 cases at last follow-up. Conclusion Induced membrane technique is a simple and reliable technique for the treatment of infectious bone defect. The technique is not limited to the size of the bone defect and the effectiveness is satisfactory.
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 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 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 role of urotensin Ⅱ(UⅡ) in the pathogenesis of asthma.Methods Lung function，differential cell count and level of UⅡin induced sputum were studied in 26 asthmatic patients in acute exacerbation and in clinical remission.Results Induced sputum UⅡ level from acute asthma was higher than that of remissed asthma [(58.88±47.38)pg/mL vs (12.69±12.78)pg/mL，Plt;0.01].Induced sputum UⅡ levels of asthma patients in acute exacerbation had a tendency to increase as disease deteriorated，which negatively correlated with FEV1% predicted (r=-0.326，Plt;0.05)，but did not with sputum total cell and neutrophil counts(Pgt;0.05).No significant difference of induced sputum UⅡ levels was found between male and female，smokers and non-smokers.Conclusion UⅡ may play a role in acute exacerbation of asthma