Adolescent ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Infant ; Infant, Newborn ; Intensive Care Units, Pediatric ; statistics & numerical data ; Lung Diseases ; epidemiology ; mortality ; physiopathology ; Male ; Prognosis ; Prospective Studies ; Respiratory Function Tests ; Survival Rate

Six DNA extraction methods and four DNA purification methods were compared and analyzed in this study to get higher quality DNA from the rhizospheric soil of Fritillaria thunbergii Miq.Results showed that higher purity DNA were harvested by pretreating the soil with 20 mmol/L EDTA(pH 7.5),then isolating soil DNA with CTAB-SDS-frozen-thawing,and further purified by agarose method.The recovery rate of this soil DNA was about 44.00 ?g/g ? 2.65 ?g/g soil,and they were qualified for the microbial diversity analysis in the rhizospheric soil of F.thunbergii Miq based on the 16S rDNA sequence.

To investigate the changes of platelet activated state and platelet activated function by trace whole blood flow cytometry (FCM), and to explore the mechanism of hemorrhage and infiltration in adults with acute leukemia, the expression percentage and changes of these expressions of CD62p and PAC-1 on platelet surface were determined by FCM of trace whole blood after platelet activated by ADP in patients with new diagnosed AL (group I), complete remission (CR, group II) and continuously complete remission (CCR, group III). Healthy adults were used as control group. The result showed that the expression of CD62p in group I and II was higher than that in control group, before and after platelet activated by ADP (P < 0.01). The expression of PAC-1 in group I was higher than that in control group (P < 0.01), the expression of PAC-1 in group II was lower than that in control group (P > 0.01), There was no significant difference in expression of CD62p and PAC-1 between group III and control group (P > 0.01), and no significant difference was found between AL group with megakaryocyte malignant pathological changes and AL group without megakaryocyte malignant pathological changes before platelet activated by ADP (P > 0.01). After platelet activated by ADP, the expression of PAC-1 in the former was lower than that in the latter (P < 0.01). It is concluded that (1) high level activated platelet in peripheral blood of AL patients show that interaction between activated platelet and leukemia cells can be one of reason resulting in widespread hemorrhage and infiltration AL patiens; (2) the decrease of number and activted function of platelet at the first stage of AL patients may be caused by malignant hyperplasia of leukemia cells and damage of megakaryopoiesis in bone marrow.
Acute Disease ; Adenosine Diphosphate ; pharmacology ; Adolescent ; Adult ; Aged ; Blood Platelets ; cytology ; metabolism ; Cell Membrane ; drug effects ; metabolism ; Female ; Flow Cytometry ; Humans ; Leukemia ; blood ; pathology ; Male ; Middle Aged ; P-Selectin ; biosynthesis ; Platelet Activation ; drug effects ; physiology ; Platelet Glycoprotein GPIIb-IIIa Complex ; biosynthesis

OBJECTIVETo investigate the effect and the possible mechanism underlying the promotional effect of Astragalus membranaceus and Panax notoginseng on the transformation of bone narrow stem cells and proliferation of EPC.

METHODThe marrow blood was collected in the patients with ischemia of lower limbs and BM-MNCs were separated and proliferated under different conditions. A. morphologic observation was performed and the ratio of CD34+ cells was measured.

RESULTThe shuttle shaped cells lined up as bunches with several round cells scattered. The ratio of CD34+ cells was significantly increased in groups treated with medium (P < 0.01) and lower (P < 0.05) dosages of A. membranaceus and medium (P < 0.01) and high dosages (P < 0.01) of P. notoginseng respectively as compared with control group.

CONCLUSIONA. membranaceus and P. notoginseng can promote the transformation and proliferation of EPC.

Antigens, CD34 ; metabolism ; Astragalus membranaceus ; chemistry ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Endothelial Cells ; cytology ; drug effects ; metabolism ; Ginsenosides ; administration & dosage ; isolation & purification ; pharmacology ; Hematopoietic Stem Cells ; cytology ; drug effects ; metabolism ; Humans ; Panax notoginseng ; chemistry ; Plants, Medicinal ; chemistry

OBJECTIVETo investigate the effects of peroral endoscopic myotomy(POEM) on esophageal dynamics in patients with esophageal achalasia.

METHODSFrom September 2011 to November 2011, 20 cases with esophageal achalasia received POEM at the Endoscopic Center in the Zhongshan Hospital of Fudan University. Pre-operation esophageal dynamics of all the patients were evaluated by high resolution manometry(HRM) system and 3 days after operation the test was repeated. Lower esophagus sphincter resting pressure(LESP), 4-second integrated relaxation pressure(4sIRP), lower esophagus sphincter relax rate(LESRR), lower esophagus sphincter length(LESL), and esophageal manometry were analyzed.

RESULTSAfter POEM, LESP decreased from(29.1±17.0) mm Hg to(14.6±4.9) mm Hg, and decrease rate was 49.8%(P<0.01). However, the decreases in LESRR and LESL were not statistically significant(P>0.05). Esophageal peristaltic contraction was absent in all the 20 patients preoperatively. After POEM, changes in the esophageal contraction were seen in 7 patients, and peristalsis was noticed but was below normal level. There were no significant changes in peristalsis in the remaining 13 patients.

CONCLUSIONPOEM can significantly reduce LESP and 4sIRP in patients with achalasia, but can not affect the contraction of the esophagus.

Adolescent ; Adult ; Aged ; Esophageal Achalasia ; physiopathology ; surgery ; Esophagoscopy ; methods ; Esophagus ; physiopathology ; surgery ; Female ; Humans ; Male ; Middle Aged ; Young Adult

Since the establishment of eight-year clinical medicine specialty, in line with the princi-ple of "eight-year consistency and fusion of the bachelor and doctor degree", the training mode of "strength-ening the foundation, focusing on quality, overall optimization, facing the clinical" has been implemented. In order to reach the standard of professional doctorate, a series of courses of professional doctorate need to be fused in limited time and designed carefully by medical schools. However, grasping proper teaching time and opportunity is particularly important for students' learning and development. By collecting the courses information of 11 medical colleges and universities offering eight-year clinical medicine specialty, we have analyzed the teaching time, methods and course categories of scientific research training courses and graduate degree courses, aiming to find the appropriate teaching program.

BACKGROUND:To date,no single material can completely meet the clinical requirements.However,the composite materials characterized by good biodegradability,biocompatibility and osteoconductivity have become a highlight of the artificial bone materials.OBJECTIVE:To synthesize the basic fibroblast growth factor (bFGF)-poly(lactic-co-glycolic acid) (PLGA) microspheres/hydroxyapatite (HA)/poly(I-lactic acid) (PLLA) porous bone scaffolds,and to observe the physicochemical properties and biocompatibility of the composite material.METHODS:The bFGF-PLGA microspheres were prepared by double emulsion method,and then six kinds of materials were made including PLLA,PLLNHA,PLLAJPLGA,PLLNHNPLGA,PLLNHA/bFGF,and bFGF-PLGA microspheres/PLLA/HA.The characterization of the materials were observed by particle size analyzer,transmission electron microscopy,X-ray diffraction,Fourier transform infrared spectrometer,microcomputer differential thermal balance,and scanning electron microscope.Toxicity of these materials and proliferation of bone marrow mesechymal stem cells seeded onto these materials were analyzed and compared.RESULTS AND CONCLUSION:The average particle size of bFGF-PLGA microspheres was about 250 nm,the average drug-loading capacity was (26.03±0.17)％,and the entrapment percentage was (90.65±2.68)％.The prepared bFGF-PLGA microspheres were spherical and had good dispersibility.In addition,all the six kinds of materials had a porous structure with similar pore diameter,in which the microspheres and particles exhibited a rational distribution.The toxic level of bFGF-PLGA microspheres/PLLNHA,bFGF/HNPLLA and HAP/PLLA was graded as 1 (with a relative survival rate ≥ 80％),indicating no obvious toxicity or slight toxicity.All these six kinds of composite materials can promote the proliferation of bone marrow mesenchymal stem cells,and the bFGF-PLGA microspheres/PLLNHA shows the best effects on cell proliferation and has good biocompatibility.

Objective To investigate the effect ofmicroRNA-101(miR-101) overexpression on proliferation and metastasis of human hepatoma HepG2 cells and the molecular mechanism.Methods HepG2 cells were divided into blank group,negative control group and miR-101 transfection group.HepG2 cell line stably overexpressing miR-101 was established by lentiviral vector.The overexpression of miR-101 was detected by chemiluminescence method.The expression of vascular endothelial growth factor (VEGF) protein was detected by Western Blot.Scratch experiments was used to analyze the cell migration and the Transwell assay was used to detect cell proliferation.Results The expression of miR-101 in HepG2 cells was significantly increased after transfection with miR-101,and there was a direct targeting relationship between miR-101 and VEGF.Compared with the negative control group,the VEGF protein level in the miR-101 transfected group was significantly down-regulated,and the difference was statistically significant (P＜0.01).Moreover,the cell scratch healing ability and invasion ability were decreased in the miR-101 transfected group.Conclusions Overexpression of miR-101 can inhibit invasion and migration of human hepatoma HepG2 cells by targeting VEGF.

OBJECTIVETo compare video-assisted thoracic surgery (VATS) and open thoracotomy (OT) on acute inflammatory responses and immunosuppression after lobectomy for early non-small cell lung cancer (NSCLC).

METHODSPresent prospective randomized study. OT or VATS lobectomy was performed in patients who met enter criteria and clinical data was collected. Plasma concentration of IL-6, IL-8 and IL-10 were measured before surgery and at postoperative day (POD) 1 and POD 3. There were 271 patients underwent lobectomy for early NSCLC, including of 133 patients in group VATS and 138 patients in group OT from January 2007 to June 2008. There were 132 males and 139 females, aging from 19 ∼ 70 years with a mean of (56 ± 8) years.

RESULTSCompared with OT group, shorter postoperative hospital stay [(8.2 ± 2.5) d vs. (9.8 ± 6.2) d, P = 0.03], lower morbidity rate (11.3% vs. 21.7%, P = 0.02) and lower increase of plasma concentration of IL-6 at POD 1 [(35 ± 25)% vs. (65 ± 43)%, P = 0.00], IL-6 at POD 3 [(14 ± 22)% vs. (55 ± 44)%, P = 0.00] and IL-10 at POD 1 [(25 ± 20)% vs. (43 ± 35)%, P = 0.00] were observed in patients of VATS group.

CONCLUSIONVATS lobectomy for early NSCLC is associated with less acute inflammatory responses and less immunosuppression when compared with OT.

Adult ; Aged ; Carcinoma, Non-Small-Cell Lung ; blood ; surgery ; Female ; Follow-Up Studies ; Humans ; Interleukin-10 ; blood ; Interleukin-6 ; blood ; Interleukin-8 ; blood ; Interleukins ; blood ; Lung Neoplasms ; blood ; surgery ; Male ; Middle Aged ; Pneumonectomy ; methods ; Prospective Studies ; Thoracic Surgery, Video-Assisted ; Thoracotomy ; Treatment Outcome ; Young Adult

Adenovirus vectors are one of the most promising gene transfer systems. They are of great value for gene therapy because these vectors achieve temporal high-level transgene expression and high gene transfer efficiency. To meet increasing needs of adenovirus vectors for gene therapy programs, parallel development of efficient, scalable and reproducible production processes is required. Perfusion cultivation of 293 cells is one of the most commonly used methods to produce adenovirus vectors and it is suitable for industrialized production specially. Experimental studies had been carried out to produce recombinant adenovirus containing the green fluorescent protein gene (Ad-GFP) by perfusion cultivation of HEK-293 N3S cells in a 5L stirring bioreactors. Perfusion rate was 1-2 volume/day. To infect the 293 N3S cells with Ad-GFP at the density of (2-4) x 10(6) cells/ ml. The time of collecting cells was 48 hours post infection. After three rounds of freeze/thaw and centrifugation, the crude viral lysates were stored at--80 degrees C until use. Then to get the Ad-GFP products by 2 x CsCl-gradient purification. The purity of the products was determined by the A260/A280 ratio and a high performance liquid chromatography (HPLC) assay. The infective titer was determined by a TCID50 assay. The culture term was 10-12 days. The infectious titer, the number of virus particle and the ratio of infectious titer to virus particle for the product were 1.0 x 10(11) IU/mL, 1.68 x 10(12) VP/mL and 6.0% IU/VP respectively. The A260/A280 ratio was 1.33, and the purity determined by HPLC was 99.2%. The cell specific productivity was around 1000 IU/cell. By perfusion cultivation of 293 N3S cells in a 5L stirring bioreactors, we established the production process for Ad-GFP, which paves a way to produce other recombinant adenovirus for gene therapy.
Adenoviridae ; genetics ; growth & development ; isolation & purification ; Bioreactors ; microbiology ; Cell Line ; Gene Transfer Techniques ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; Humans ; Kidney ; cytology ; virology ; Recombinant Proteins ; biosynthesis ; genetics ; Recombination, Genetic ; Virus Cultivation ; instrumentation ; methods