Objective To observe the effects of dynamic pressure for the ability of endothelial progenitor cells (EPCs) to form blood vessels, when EPCs seeded into DBM with load. Methods Use the Ficoll density gradient centrifuge combined with difference-speed adherence screening method to separate MNCs from rat bone marrow. Identify the induced EPCs by means of immunohistochemistry and immunofluorescence. Through the organization of fixed, defatted, decalcified and other steps use of spine vertebral body,demineralized bone matrix (DBM) samples of pig were prepared in vitro. Divided scaffolds into two groups A and groups B. Induced EPCs were seeded into DBM. The cell-seeded scaffolds of groups A were dynamically loaded in compression using a sine wave at 1 Hz, 5% strain in the media-filled chamber for 4 h on days 5 of culture. and cell-seeded scaffolds of groups B were cultured directly without any load. Both of two groups were cultured two weeks. Then the ability of EPCs to form blood vessels was observed. Primer desig;Extract total RNA from cells with Trlzol; Reverse transcription reaction; PCR. Results Two groups of cells in HE staining and fluorescent staining showed the formation of vascular bundles. There were formation of blood vessels. It was obvious that the formation in group A was more than that in group B. Test the mRNA expression of vWF and Flk-1 during the EPCs differentiationby RT-PCR. Group A was significantly stronger than that of group B. Conclusion When DBM combines together with EPC, it has become organization engineering bone, then with press on it, the bone graft has been vascularized, so it has clinical application on the direction of repair bone defect.

BACKGROUND: Ventilation dysfunction caused by bronchomalacia induced bronchostenosis following high-risk heart transplantation is an acute clinical disease, which seriously impairs the function of transplant heart. The case of emergency bronchial stent placement following heart transplantation with high-risk multi-complication has not been reported yet.OBJECTIVE: To investigate the curative effect of emergency stent placement for worse left main bronchial malacia, stenosis and collapse following orthotopic heart allotransplantation.DESIGN: A case analysis.SETTINGS: National Ministry of Health Transplantation Engineering and Technical Research Center, the Third Xiangya Hospital, Central South University; Department of Cardiosurgery, Zhongshan Hospital affiliated to Xiamen University.PARTICIPANTS: An 18-year-old female patient with dilated cardiomyopathy accompanied by moderate to severe pulmonary artery hypertension, who sequentially carried out orthotopic heart allotransplantation, was selected from the Department of Cardiosurgery, Zhongshan Hospital affiliated to Xiamen University in April, 2004. She had suffered from dilated cardiomyopathy for 15 years, and the mean pulmonary artery pressure (MPAP) was 50-51 mm Hg, she was also accompanied by left main bronchial malacia, stenosis and collapse, mixed (mainly central-) sleep apnea syndrome, left inferior pulmonary sequestration, right emphysema, and rheumatoid arthritis for half a year.METHODS: After heart transplantation, bronchus inflammation, congested edema aggravated the severity of bronchial malacia, stenosis and collapse, tenosis reduced to 4/5, and led to obstructive type of ventilation, and the patient was also accompanied by supraventricular tachycardia, ventricular extrasystole, and hypofunction of transplant cardiac systolic function (peak E＜peak A, ejection fraction reduced to 40%, inharmonious motion of ventricular wall). Attempted with inotropic agents and ventilator/support were not relieved, which resulted in the aggravation of illness. In order to improve the post-transplant cardiac function, to relieve bronchial collapse and stenosis, and correct the obstructive type of ventilation, an emergency bronchial stent placement surgery was carried out on the sixth day after heart transplantation. Under monitoring of electrocardiogram (EGG) and percutaneaous oxygen saturation (SpO2), patient was awake and in supine to relieve left main bronchial stenosis with a nickel-titanium shape memory alloy stent (Diameter: 12 mm; length: 20 mm) by D20 fiberoptic bronchoscope. Fibrobronchoscopy was used to observe the proximal end of bronchostenosis and set the proximate location mark by using video fluoroscopy; the patency of distal end was explored by stricture, and set the distal location mark; guidewire was inserted into working path of bronchofibroscope and led through the stricture; then loaded the Ni-Ti stent on a special placement apparatus, and led in bronchial stent implantation apparatus along guidewire. When targeting well, the stent was slowly released and adjusted properly. When it was completely released, the stent implantation apparatus was drawn out. Bronchofibroscope was performed postoperatively to observe the adherence of stent; immediately photographed to observe its unfolding. Synchronized intermittent mandatory ventilation (SIMV) was given postoperatively as supportive treatment.MAIN OUTCOME MEASURES: Ameliorations of the cardiac and pulmonary functions of the patient.RESULTS: ①Carbon dioxide retention and hypercapnia were remarkably improved as compared with those preoperatively; hypertensive pulmonary vascular disease was alleviated gradually, and MPAP reduced to 30 mm Hg. One week later, re-examination of bronchofibroscopy was carried out, and the results showed that bronchi of left upper lobe, lingual lobe as well as left lower lobe could be seen distinctly, mucous membrane had slightly congested edema, and lumens were unobstructed.②Supraventricular tachycardia and premature ventricualr contraction disappeared, and the transplant cardiac function recovered well (peak E ＞ peak A, ejection fraction 70%, FS41%), and the heart rate fluctuated at 100-110 beats per minute. ③The chest-radiography and CT postoperatively indicated the relief of left main bronchial stenosis. When the ventilation function of the patient was improved, the parameters of breathing machine were reduced gradually, and replaced by low-flow oxygen. There was no recurrence of obstructive ventilatory disorder. The sleep apnea syndrome of the patient was moderated.CONCLUSION: Emergency treatment with stent placement for bronchial malacia, stenosis and collapse occurring after orthotopic heart allotransplantation cAN improve ventilation dysfunction caused by bronchial malacia and stenosis,and increase the survival rate of heart transplantation.

Amyotrophic lateral sclerosis (ALS) is the most common motor neuron disease. At present, no definite ALS biomarkers are available. In this study, exosomes from the plasma of patients with ALS and healthy controls were extracted, and differentially expressed exosomal proteins were compared. Among them, the expression of exosomal coronin-1a (CORO1A) was 5.3-fold higher than that in the controls. CORO1A increased with disease progression at a certain proportion in the plasma of patients with ALS and in the spinal cord of ALS mice. CORO1A was also overexpressed in NSC-34 motor neuron-like cells, and apoptosis, oxidative stress, and autophagic protein expression were evaluated. CORO1A overexpression resulted in increased apoptosis and oxidative stress, overactivated autophagy, and hindered the formation of autolysosomes. Moreover, CORO1A activated Ca²⁺-dependent phosphatase calcineurin, thereby blocking the fusion of autophagosomes and lysosomes. The inhibition of calcineurin activation by cyclosporin A reversed the damaged autolysosomes. In conclusion, the role of CORO1A in ALS pathogenesis was discovered, potentially affecting the disease onset and progression by blocking autophagic flux. Therefore, CORO1A might be a potential biomarker and therapeutic target for ALS.
Mice ; Animals ; Amyotrophic Lateral Sclerosis/pathology* ; Calcineurin/metabolism* ; Motor Neurons/pathology* ; Microfilament Proteins/metabolism* ; Cytoskeletal Proteins/metabolism*