OBJCETIVETo investigate the method of separation of culture of bone microvascular endothelial cells (BMECs) of human femoral head in vitro.

METHODSFrom October 2013 to January 2014,15 femoral heads without pathologic change from patients resected during hip replacement were selected involving 2 males and 13 females with a mean age of 71.2 years old ranging from 38 to 92. Cancellous bone in femoral head was bited into broken bone grain and transfered into medium in aseptic contidion. Cells were isolated by the methods of enzymic digestion and density gradient centrifugation,purified by differiential attachment. The characteristics of cells was observed by inverted microscope. vWF and CD31 immunofluorescence analysis was applied for identification of cells.

RESULTSThe number of cells was positively correlated with patients' age after 24 hours in primary culture. The older patients had the less cells numbered. After 4 to 5 days' culture, primary cells appeared short spindle,polygon shaped and cobblestone-like morphology. After 7 to 10 days' culture, primary cells proliferated densely, became fusion, arranged in swirl, and contact inhibition appeared significantly. Immunofluorescence staining revealed the cells were 100% positive for vWF and CD31, and it showed that the cultured cells were BMECs.

CONCLUSIONIt was a simple, steady, effective method with good reproducibility, by which highly purified human BMECs can be obtained.

Adult ; Aged ; Aged, 80 and over ; Cell Culture Techniques ; Cell Proliferation ; Cell Separation ; methods ; Cells, Cultured ; Endothelial Cells ; cytology ; Female ; Femur Head ; blood supply ; Humans ; Male ; Microvessels ; cytology ; Middle Aged

Objective To assess the level of radiation exposures of operators in three typical types of interventional fluoroscopic procedures.Methods Alderson Radiation Therapy (ART) phantom was used to stimulate the practices of diagnosis and therapy using TLDs for dose measurement.The radiation exposures of eye lens, neck, and breast were measured when the lead shielding of machine was on/off and the equivalent dose and effective dose to the eye lens were estimated.Results Radiation exposure of head was obviously reduced by 85% -90% when the lead shielding was on.The doses in different procedures were different.In cerebral angiography the dose equivalent of eye len was the highest in the three procedures.The annual effective dose for the operators was smaller in peripheral vascular interventions than that in cardiovascular interventional therapy and that in cerebral angiography.Conclusions The operators involved in intervention will receive an annual effective dose of less than 20 mSv as recommended by the ICRP under the protection conditions provided by the current study, except for eye lens.Attention should be paid to the protection of the eyes of operators.

Objective To study the radiation dose level and dose distribution of leakage and scattering radiation in a helical tomotherapy (TOMO) room,and to identify the dose distribution characteristic which is different from the conventional radiotherapy so as to provide scientific data for the radiation protection of TOMO.Methods A helical tomotherapy facility typed Tomotherapy Hi-Art was employed as the radiotherapy device.The air kerma from leakage radiation and scattering radiation distributed on the couch plane and in TOMO room were detected mainly by GR-200A TLDs of LiF(Mg,Cu,P),while 100 Gy were being accumulated under the condition of simulation radiation treatment.Then,the leakage radiation ratio and scattering-leakage radiation ratio (the ratio of site radiation dose to isocenter standard output dose) were calculated.Results The radiation level and dose distribution of leakage radiation and scattering radiation in the TOMO treatment room were in bilateral symmetry to the rotation isocenter and the treatment couch longitudinal axis,and the radiation level in front of the radiotherapy device was significantly higher than that of the rear of the TOMO equipment.In the plane of treatment couch,the maximum leakage radiation ratio at the position 100 cm away from the isocenter was only 1.3 × 10-4,and the mean value of the leakage radiation ratio at the position 300 cm away from the isocenter was lower than 2.0 × 10-5.The scattering radiation at 200-300 cm away from the isocenter was accounted for 25％-30％ of the leakage radiation from the responding sites,and the indoor scattering radiation decreased faster than leakage radiation with the distance from the isocenter.Conclusions The radiation level of leakage and scattering in a TOMO room is significantly lower than that in a conventional radiotherapy room.

Objective To prepare an attenuated Listeria vaccine Lmdd-LMP2A expressing the Ep-stein-Barr virus latent membrane protein 2A ( EBV-LMP2A) and evaluate its specific anti-tumor effects on nasopharyngeal carcinoma.Methods The gene fragment encoding EBV-LMP2A was amplified by PCR analysis and then subcloned into the shuttle vector p1565.PCR restriction enzyme digestion and DNA se-quencing were performed to identify the recombinant shuttle vector p1565-LMP2A.The p1565-LMP2A vector was then transformed into competent strains of the attenuated Listeria monocytogenes ( Lmdd) .The recombi-nant attenuated Listeria vaccine strain Lmdd-LMP2A was verified by Western blot assay.The histological sections of spleen and liver tissues were stained by Haematoxylin and eosin ( H&E) for analysis of inflamma-tion.A tumor-bearing HLA-A2 transgenic mouse model was established by subcutaneous injection of CNE-1/HLA-A2/LMP2A nasopharyngeal carcinoma cell line.The prepared Lmdd-LMP2A vaccine was inoculated into the mice via tail intravenous injection for the evaluation of specific CTL induction and the in vivo anti-tumor effects.Results The shuttle vector p1565-LMP2A and the recombinant attenuated Listeria vaccine strain Lmdd-LMP2A with stable expression of LMP2A protein were successfully constructed.The immunized mice showed mild inflammations with no structural damage and necrosis as indicated by H&E staining.The growth of tumors in tumor-bearing HLA-A2 transgenic mice was significantly inhibited by the immunization of Lmdd-LMP2A vaccine as compared with mice without inoculation.The survival time of mice was prolonged with the immunization of Lmdd-LMP2A vaccine.Conclusion The prepared attenuated Listeria vaccine Lm-dd-LMP2A showed specific anti-tumor effects with the safety advantage, suggesting the possibility of using it for anti-tumor therapy in clinic.

Sanjie Zhentong capsules were scanned by using a near infrared spectra probe with different drug mass fraction and the spectral information of capsule shells and contents in it were obtained. Then partial least squares (PLS) models were developed for the prediction of mass fraction of Fritillariae Thunbergii Bulbus and Resine draconis in Sanjie Zhentong capsules. The correlation coefficient (r9c)) and root mean standard error( RMSEC) of 0.949 5, 0.958 2 and 4.742 4, 4.135 7. The models obtained correlation coefficient (r(v)) of 0.919 2, 0.936 7 and root mean square error (RMSECV) of 6.158 9, 5.037 3 respectively in the training set. The paired T test analysis of statistics showed that there were no significant difference between predictive values and measure values. The established models reflected a strong prediction performance and can meet the needs of the production.
Capsules ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Least-Squares Analysis ; Spectroscopy, Near-Infrared ; methods

5.0mg/L were randomly divided into LMWH treatment group and low molecular dextran treatment group with 20 patients in each group.The patients in LMWH group were treated with 0.3ml LMWH subcutaneous injection in abdominal wall in every 12h for 1-4 d.The patients in low molecular dextran group were treated with 500ml low molecular dextran plus 20ml danshen root,intervenous drop infusion for 1-7d.Results The D-Dimer blood serum level in the gestational late period was significantly higher than that of nongravida group(P

Objective To investigate the radiation dose levels to the adults examined from diagnostic exposure in Beijing. Methods The radiation doses to the examined individuals were measured by using individual diagnostic radiology equipments in 30 random hospitals from a total of 10 districts and suburban areas, including 1 182 samples of X-ray photography,542 samples of mammography and 410 samples of CT examination. Results 2 134 samples were measured in this study. The dose ranges of X-ray photography, CR, and DR were 0?4 -24?1, 0?3 -13?9 and 0?1 -15?9 mGy, respectively. The average dose range of glandular breast was 0?3-5?4 mGy. In 410 CT samples the value of CTDIw , CTDIvol and DLP were 28?1 - 96?3 mGy, 7?0 - 23?4 mGy, and 162?2 - 898?1 mGy·cm, respectively. Conclusions Several dose levels from diagnostic examination were higher than guidance level for medical exposure in GB 18871-2002,which should be noted.

Objective:To compare the safety and effectiveness of two minimally invasive approaches for multi-vessel coronary revascularization.Methods:From August 2014 to February 2017,70 consecutive patients who underwent minimally invasive coronary artery bypass grafting in Peking University Third Hospital were randomly divided into two groups.In one group,40 patients underwent staged-hybrid coronary revascularization (staged-HCR) treatment;in the other group,30 patients underwent minimally invasive total arterial revascularization with bilateral internal thoracic artery (BITA).In staged-HCR group,the patients underwent minimally invasive direct coronary artery bypass grafting (MIDCAB) and percutaneous coronary intervention (PCI) procedure for treatment of multi-vessel disease.In BITA group,the patients underwent total arterial coronary artery bypass grafting with composite "Y" BITA graft.Preoperative and postoperative data of the two groups,including postoperative blood usage,mechanical ventilation time,domiciling duration in intensive care unit (ICU),major adverse cerebral and cardiovascular event (MACCE),and postoperative coronary angiography results were compared,in order to evaluate the safety and effectiveness of these surgical approaches.Results:The preoperative characteristics of 70 patients in the two groups showed no significant difference.All the patients underwent successfully,elective minimally invasive multi-vessel coronary artery bypass grafting as scheduled preoperatively.Postoperative result showed the patients in staged-HCR group took advantages in less postoperative mechanical ventilation time [Staged-HCR group (11.2 ± 8.7) h vs.BITA group (18.3 ± 9.1) h,P =0.013],shorter domiciling duration in ICU [Staged-HCR group (26.29 ± 4.05) h vs.BITA group (44.74 ± 28.75) h,P =0.022],and less total drainage [Staged-HCR group (695.57 ± 250.46) mL vs.BITA group (1 103.26±547.44) mL,P =0.03] than the patients in the group of minimally invasive total arterial revascularization with BITA.Postoperative in hospital coronary angiography showed satisfactory graft patency rates in both groups [97.5％ in Staged-HCR group vs.97.8％ in BITA group].No MACCE occurred in both groups during hospitalization.Conclusion:Staged-HCR is a feasible method for the treatment of multi-vessel revascularization involving right coronary artery.Minimally coronary revascularization with BITA is associated with superior long-term graft patency and it's recommended for patients who could not tolerate dual-antiplatelet therapy.This study shows that both minimally invasive surgical approaches are safe and effective for treatment of patients with multi-vessel coronary artery disease.

OBJECTIVENumber and function of endothelial progenitor cell (EPC) and coronary artery lesion in Kawasaki disease (KD) model were evaluated to investigate therapeutic efficacy of granulocyte colony-stimulating factor (G-CSF).

METHODC57BL/6 mice were injected with L. casei cell wall extract (LCWE); 48 mice were divided into 3 groups randomly: KD model group; G-CSF treated model group and control group, 16 in each. G-CSF was subcutaneously injected from day 5 to day 9 after injection of LCWE. Coronary artery lesion, number of circulating EPC and the function of bone marrow EPC were evaluated.

RESULTIn model group, inflammatory infiltration was found around coronary artery at 14 days. The number of circulating EPC was significantly decreased in model group (0.017% ± 0.008%) compared to control (0.028% ± 0.007%) (t = 2.037, P < 0.05). Disruption of elastin was consistently observed at 56 days. Stimulated by G-CSF, inflammatory infiltration was found around the coronary artery at day 14, while the number of circulating EPC (0.042% ± 0.015%) was increased significantly compared to models (t = 4.629, P < 0.05). At the day 56, the number of circulating EPC was decreased slightly (0.029% ± 0.012%), but still higher than the model group (t = 2.789, P < 0.05), and have no significant difference compared to controls (P > 0.05). Furthermore, there was no elastin disruption in the G-CSF group. In model group, bone marrow EPC's proliferation ability of absorbance (A value) was 0.38 ± 0.09 in thiazolyl blue assay, less than controls (0.61 ± 0.14, P < 0.01). Adhesion and migration function were down-regulated compared to controls [(3.1 ± 0.6) cells/HPF and (3.3 ± 0.6) cells/HPF vs. (6.4 ± 1.2) cells/HPF and (6.2 ± 0.5) cells/HPF, both P < 0.01]. In the G-CSF treated group, proliferation ability (A 0.58 ± 0.10), adhesion [(6.17 ± 1.13) cells/HPF], migration [(6.29 ± 0.42) cells/HPF] function were increased significantly compared to the model group (P < 0.01).

CONCLUSIONG-CSF can up-regulate EPC number and function to prevent coronary artery lesion in mice model of KD.

Animals ; Coronary Vessels ; drug effects ; pathology ; Disease Models, Animal ; Endothelial Cells ; cytology ; drug effects ; Flow Cytometry ; Granulocyte Colony-Stimulating Factor ; administration & dosage ; therapeutic use ; Male ; Mice ; Mice, Inbred C57BL ; Mucocutaneous Lymph Node Syndrome ; blood ; drug therapy ; pathology ; Random Allocation ; Stem Cells ; cytology ; drug effects ; Up-Regulation

The rhizome of Panax japonicus var. major have been used as the natural medicinal agent by Chinese traditional doctors for more than thousand years. Most of the therapeutic effects of P. japonicus var. major had been reported due to the presence of tetracyclic or pentacyclic triterpene saponins. In this study, Illumina pair-end RNA-sequencing and de novo splicing were done in order to understand the pathway of triterpenoid saponins in this species. The valid reads data of 15. 6 Gb were obtained. The 62 240 unigenes were finally obtained by de novo splicing. After annotation, we discovered 19 unigenes involved in ginsenoside backbone biosynthesis. Additionally, 69 unigenes and 18 unigenes were predicted to have potential function of cytochrome P450 and UDP-glycosyltransferase based on the annotation results, which may encode enzymes responsible for ginsenoside backbone modification. This study provides global expressed datas for P. japonicus var. major, which will contribute significantly to further genome-wide research and analysis for this species.
Gene Expression Profiling ; Panax ; genetics ; Saponins ; biosynthesis ; Sequence Analysis, RNA