AIM: To study the endothelial progenitor cell markers and biological characteristics of human umbilical cord blood CD133+ cells isolated by immunomagnetic bead (IMB) method. METHODS: The experiment was carried out in the Laboratory of Burn Surgery, Xijing Hospital of the Fourth Military Medical University of Chinese PLA from November 2002 to August 2003.①CD133+ cells were enriched from human umbilical cord blood of placenta through spontaneous delivery or cesarean delivery (offered by Department of Gynecology and Obstetrics, Xijing Hospital of the Fourth Military Medical University of Chinese PLA), with the informed consents of parturients.②Human umbilical cord blood was anticoagulated by ACD-B and then diluted, 7 mL of cord blood combined with 3 mL lymphocyte isolation liquid were centrifuged for 30 minutes at 2 100 r/min. Low-density mononuclear cells coloring buffy were selected to wash off platelet and inoculated for 30 minutes by the IMB and FITC-CD133 antigen at room temperature to wash excessive antigens. The FITC-CD 133-coated IMB was added into mononuclear cells which were cultured for 30 minutes at room temperature and placed in magnetic field for 1 minute to remove the cell suspension. IMB-cell isolation liquid was supplemented to release CD133+ endothelial progenitor cells, which were cultured with EGM-2MV media in dishes coated with collagen I. And 24 hours later those unadherent cells were detached, culture liquid was removed half after 3 hours, and total culture duration was 3-4 weeks.③The morphologic change of CD133+ cells isolated with IMB were observed; The percentage of CD133+ cells in cord blood monocytes and sorting efficiency of IMB were analyzed by fluorescence activated cell sorting (FACS). The cells cultured in vitro were identified by fluorescence immunoassay and enzymo-histochemistry while transmission electron microscope (TEM) was used to observe the W-P body of mature vascular endothelial cells. RESULTS: ①Morphologic observation of CD133+ cells culture in vitro: The adherent cells expanded pseudopods on days 4 and 5, and grew rapidly in clonal manner 7 days later. The cells showed a typical spindle-shaped morphology during 2-3 weeks and cluster areas of cobblestone-like cells after 2-week culture and confluence.②Isolation rate of CD133+ cells: The percentage of CD133+ cells in cord blood monocytes was 0.91%. And after IMB sorting, the percentage of CD133+ cells was 85.52%.③On the fifth day, few cells were positive for CD133 while most cells were positive for CD 34 and vWF; On the tenth day, all cells were negative for CD133 and most cells were still positive for CD 34 and vWF.④The cells after 10-day culture had typical W-P body in TEM. CONCLUSION: Immunomagnetic sorting can efficiently select enriched CD133+ endothelial progenitor cells from human umbilical cord blood. Our data of CD133, CD34, vWF antibodies and TEM support the hypothesis that endothelial progenitor cells may contribute to mature vascular endothelial cells.

To prepare an experimental caprine model of tibia bone diaphyseal defect used in tissue engineering and defect repair with the tissue engineering method, 27 Chinese caprines were divided into 3 groups: blank, control, and test. The 20mm left tibia diaphyseal defect of each caprine was made and fixed with plates. The blank group was not filled with coral hydroxyapatite (CHAP) or bone marrow stroma cell (BMSc). The control group and the experimental group were filled with CHAP and CHAP/BMSc respectively. The results of three groups were evaluated by X ray examination, optical density index of X ray film and histology 4, 8, 12 weeks after operation. The biomechanical characteristics of the specimens of the CHAP group and the CHAP/BMSc group were tested by three point bending test 12 weeks after operation.The results showed that the optical density indices of X ray film of the blank group were not significantly different 4, 8, 12 weeks after operation and indices of the CHAP group were significantly smaller than those of the CHAP/BMSc group ( P

Corneal neovascularization is a significant and sight-threatening complication of many ocular surface disorders, and may cause corneal sear and rejection reaction after corneal grafting. Recent studies have revealed that vascular endothelial growth factor (VEGF) plays an important role in corneal neovascularization, and inhibition of VEGF has become a main strategy for treatment of corneal neovascularization. This article reviews the research progress in inhibition of corneal neovascularization by anti-VEGF therapy.

Objective To analyze and compare the ultrasound elastography (UE) features of various types of cervical tuberculous lymphadenopathy (CTL), and to investigate the value and limitation of UE in diagnosis of CTL. Methods A total of 70 CTL patients confirmed by pathology or diagnostic treatment who were admitted to Shanghai Pulmonary Hospital affiliated to Tongji University from Jul. 2017 to Mar. 2018 were enrolled, and the images of conventional ultrasound and UE of the largest lymph nodes in each patient were retrospectively analyzed. According to the conventional ultrasound classification criteria of tuberculous lymphadenopathy, the lesions were divided into acute inflammatory type (type), caseous necrotic type (typeⅡ), cold abscess type (type III) and healing calcification type (type ). The elasticity score and strain ratio of various types of CTL lesions were summarized and compared. Results There were 24 (34.29%) typeCTL lesions, 28 (40.00%) typeⅡ, 12 (17.14%) type III, and 6 (8.57%) type . The hardness of CTL lesions of typeand typeⅡ was high, with the elastic score being 3-4 (47/52, 90.38%) and the strain ratio being 3-5; the hardness of type III was the lowest, with the elastic score being 1-2 (10/12, 83.33%) and the strain ratio2; the hardness of type was the highest, with the elastic score being 4-5 (6/6, 100.00%) and the strain ratio5. There were significant differences in elastic score and strain ratio among the four type CTL lesions (H＝30.756, F＝23.177; both P0.001). Pairwise comparison showed that except for the differences between typeand typeⅡ lesions, there were significant differences in elastic scores and strain ratios between the four type CTL lesions (P0.05). Conclusion The value of UE in the diagnosis of CTL is limited. It may play some roles in determining whether there is an obvious granuloma formation in typelesions and the tension of type III lesions.

Objective To investigate the value of intrathoracic contrast-enhanced ultrasound in the treatment of pleural effusion. Methods Forty-four patients in our hospital were selected; they had stopped drainage during closed thoracic drainage and had residual pleural effusion determined by pathogenetic condition. The resistance in the catheter during normal saline flushing, whether the effusion could be drained and the effusion traits, and the enhancement of catheter, chest wall soft tissue and pleural cavity after contrast-enhanced ultrasound were all observed. Clinical and ultrasound examination was used to judge whether the catheters were out of the effusion cavity and the accuracies of the two examination methods were compared. Results When the catheter was located in the effusion cavity, contrast-enhanced ultrasound showed a linear enhancement in the catheter and a slow enhancement in the effusion cavity; when the catheter was removed, contrast-enhanced ultrasound showed a short linear enhancement in the catheter, a limited area of enhancement in the chest wall soft tissue and no enhancement in the effusion cavity. In clinical judgment, 8 catheters were located in the effusion cavity, and 36 cases were removed. In ultrasound judgment, 29 catheters were located in the effusion cavity, 14 cases were removed, and 1 case could not be judged. Taking whether there is still liquid extraction 24 h after saline flushing or drug injection as the standard of catheter removing, we found that the accuracy of contrast-enhanced ultrasound in determining catheter removing was significantly higher than that of the clinical judgment (97.73% [43/44] vs 50.00% [22/44], P0.01). Conclusion Intrathoracic contrast-enhanced ultrasound can accurately judge the position of catheters and the internal condition of the effusion cavity. It provides a basis for further treatment of pleural effusion.

Disaster Planning ; organization & administration ; Emergencies ; Emergency Medical Services ; organization & administration ; Humans ; Public Health ; methods

Objective To investigate the application effect of scene exercise training model of 6 emergency disease entities in the junior nurses training .Methods Thirty‐six newly employed nurses during 2010-2013 were performed the scene exercise train‐ing of six disease entities .Before and after training ,the the forms of questionnaire investigation ,written examination and implemen‐tation were adopted to survey and analyze the post competence ,training attitude ,theoretical knowledge ,operation skill ,single time for help and participating doctor′s satisfaction on the coordinate ability .Results After training ,the aspects of post competence , training attitude ,theoretical knowledge ,operation skills ,single time for help and participating doctor′s satisfaction on the coordinate ability in junior nurse were significantly improved ,the differences between before and after training had statistical significance (P<0 .01 or P<0 .05) .Conclusion Using the scene exercise training model of 6 emergency disease entities in junior nurses training has active significance for increasing the overall quality of junior nurses .

Objective To construct an immortalized rat astrocyte strain expressing enkephalin which can be regulated by doxycycline.Methods pRevTet-On and recombinant plasmid pRevTRE/hPPE were transfected into packaging cell PT67 respectively by standard lipofectamine methods.The supernatant containing RevTet-On virus was used to infect immortalized rat astrocyte strain(IAST)and RevTRE/hPPE virus was used to infect the IAST/Tet-On cell.Immortalized rat astrocyte strain that stably expressed Tet-regulated enkephalin was established. hPPE gene expression level of IAST/Tet-On/hPPE cell strain was detected by real time-PCR,immuno- cytochemistry and radio-immunoassay.Results Real time-PCR,immuno-cytochemistry and radio-immunoassay confirmed the level of enkephalin expression was regulated by doxycycline in a dose-dependent manner.Conclusion An immortalized rat astrocyte strain which secrets enkephalin as controlled by doxycycline has been constructed successfully.

Objective The effect of electronic cigarettes (e-cigarettes) on smoking reduction,cessation and human health remains controversial,meanwhile,studies tocused on e-cigarettes are limited.We conducted this meta-analysis in purpose of helping figuring out the controversy.Methods PubMed, Medline,Embase,Cochrane library,CNKl and Wanfang Databases were searched for articles published up to May 2014.All studies discussing the relationship between e-cigarettes and smoking reduction,smoking cessation or human health were included.The fixed or random-eftect model was used to pool the relative risk(RR).Outcome 4 articles were included based on the inclusion and exclusion criteria,there were totally 7 002 subjects.Meta-analysis indicated that there was significant difference in e-cigarettes and smoking reduction,RR=1.88,95％ CI:1.07-3.32,P=0.09.The Meta-analysis showed that e-cigarettes were available in smoking cessation.RR=1.55,95％ CI:1.24-1.95,P=0.001.Conclusion Tiffs meta-analysis indicated that e-cigarettes were helpful in smoking cessation.Nonetheless,the articles included were too few to credibly support the outcome.Hence,more high-quality,randomized blind and large sample experiments are needed to verify the results.

Objective To observe the effects of human retinal pigment epithelium conditioned medium(HRPE-CM) on the biological characteristics of human retinal stem cells(HRSCs). Methods HRSCs were exposed to HRPE-CM and cultivated in three different cultures,including the control,epidermal growth factor(EGF) + basic fibroblast growth factor(bFGF) and HRPE-CM.Cell counting was performed to explore the effects of different culture media on the proliferation of HRSCs,and their properties as neural stem cells were further identified. Results Compared with control group,HRPE-CM significantly promoted the proliferation of HRSCs(P