Abdominal Pain ; etiology ; Capsule Endoscopy ; Child ; Gastrointestinal Hemorrhage ; etiology ; Humans ; Intestinal Diseases ; complications ; diagnosis ; etiology ; Intestine, Small ; blood supply ; pathology ; Male ; Telangiectasis ; complications ; diagnosis ; pathology

Objective　To observe some biological features of bone marrow mesenchymal stem cells and explore the best conditions for isolatin g and culturing in vitro. Methods　Common cell culture techn ique, light and electron microscopy were used to study the effects of the growth , proliferation, morphology of the bone marrow mesenchymal stem cells in differe nt adherent time, concentration of serum and cell density. Results　The best culture condition in vitro for growth was 4-24 hours adherent time, 5%-10% fetal bovine serum, (4-8)×104/ml cell density. The cells were sp indle in shape and had a strong ability of proliferation. The time for cell duplication was 3 to 4 days. The cells showed the characteristics of stem cell s in electron microscope. Conclusion　The best condition for iso lation and culture of bone marrow mesemchymal stem cells was successfully establ ished and some biological features were obserred. It found a base for further in vestigation and using of mesenchymal stem cells.

OBJECTIVETo evaluate the effect and results of short and medium periods of follow-up of percutaneous balloon pulmonary valvuloplasty for critical pulmonary stenosis of neonates and infants under 6 months of age.

METHODSBetween January 2002 and December 2008, 34 consecutive patients aged from 13 to 175 days with critical pulmonary valvular stenosis underwent percutaneous balloon valvuloplasty. Patients records, catheterization data, angiograms and echocardiograms were reviewed. Patients were followed up for 6 months to 4 years (mean 25.5 months) by means of clinical examination and Doppler echocardiography.

RESULTSThe pulmonary valvuloplasty was accomplished in 32 (94%) of 34 attempts. Immediately after dilation, right ventricular systolic pressure (RVSP) decreased from (96 ± 28) mm Hg (1 mm Hg = 0.133 kPa) (49 ± 20) mm Hg (P < 0.01), the transvalvular peak to peak systolic gradient (ΔP) decreased from (89 ± 25) mm Hg to (25 ± 12) mm Hg (P < 0.01), and the right ventricular/aortic systolic pressure ratio decreased from 1.2 ± 0.5 to 0.7 ± 0.3 (P < 0.01). One patient died because of cardiac tamponade following rupture of the pulmonary valve annulus, 2 patients developed pericardial effusion, 3 patients had infundibular spasm, 3 patients had a pre-dilation by small balloon and 1 patient had weakened femoral artery pollex. After a follow up period of 6 months to 4 years 3 of 31 patients lost to follow-up. Repeat valvuloplasty was performed in 5 patients (3 neonates), no patient required surgery, and the other 23 patients did not undergo further intervention, a mean peak systolic Doppler gradient of (20 ± 13) mm Hg was found and no significant pulmonary regurgitation was seen.

CONCLUSIONSPercutaneous balloon pulmonary valvuloplasty was effective and safe for the treatment of critical pulmonary stenosis of neonates and infants under 6 months of age with good short and medium term results.

Catheterization ; Female ; Follow-Up Studies ; Humans ; Infant ; Infant, Newborn ; Lost to Follow-Up ; Male ; Pulmonary Valve ; Pulmonary Valve Stenosis ; therapy ; Treatment Outcome

Objective To study the effect of essential oil from Angelica sinensis(EOAS) on neuronal apoptosis induced by ischemia-reperfusion injury.Methods The high differentiated PC12 cells were divided into blank group,model group,control group (10 μmol · L-1 edaravone)and large-,medium-and small-dose expreimental groups(EOAS 25.00,12.50,6.25 μg · mL-1).Except blank group,the remaining groups were treated with sugar-free medium containing 10 mmol · L-1sodium dithionite (Na2S204) for 1 h,reoxygenation for 48 h.The cell proliferation was measured by methyl thiazolyl tetrazolium(MTT) assay.The activity of superoxide dismutase (SOD) and lactate dehydrogenase (LDH),contents of malondialdehyde (MDA) were measured by kit.The apotosis rate and the morphological apoptotic characteristics of cells were observed by flow cytometry and acridine orange/propidium iodide (AO/PI) double staining respectively.The relative activity of caspase-3 were examined with colorimetric assay.The expression of p-ERK1/2 proteins was measured by Western-blot.Results Compared with the blank group,the cell viability in the model group was (67.4 ±0.10)％ with significantly(P ＜0.01).Compared with the model group,the cell viability in the large-dose experimental group and control group were (86.2 ± 0.10)％,(94.5 ± 0.05)％ with significantly (P＜0.05,P ＜ 0.01).Compared with the blank group,the LDH and MDA content in the model group were (912.53±16.71)U · L-1and(9.05 ±0.25)μmol · L-1while SOD level was (12.53 ±0.29)U · mL-1with significantly(all P ＜0.01).Compared with the model group,the LDH and SOD level in the large-dose experimental group were (565.61 ± 11.72) U · L-1 and (12.53 ± 0.29) U · mL-1 with significantly (P ＜ 0.05,P ＜ 0.01) while MDA content in the large-,medium-dose experimental groups were (3.32 ± 0.68),(5.79 ± 0.68) μmol · L-1 with significantly(all P ＜0.01).The absorbance(A) in model group and large-,medium-and small-dose expreimental groups were 0.75 ± 0.06 and 0.10 ± 0.02,0.16 ± 0.03,0.49 ± 0.04.Compared with the model group,the difference had significantly(P ＜0.05,P ＜0.01).Compared with the blank group,the apoptosis rate in the model group was (31.17 ± 2.44)％ with significantly(P ＜ 0.01).Compared with the model group,the apoptosis rate in the large-,medium-dose experimental groups were (4.57 ± 0.32) ％,(5.93 ± 0.81) ％ with significantly (all P ＜ 0.01).Compared with the model group,p-ERK1/2 proteins of large-dose experimental group were up-regulated significantly (P ＜0.01).Conclusion The EOAS could inhibit the apoptosis of neurons after ischemia-reperfusion by activating ERK signaling pathway.

Objective:To improve the genetic stability of HBV gene in transgenic mice.Methods:HBV transgenic mice were bred by backcross and double cross.The HBV gene expression and replication were studied with real-time PCR,ELISA and chemiluminescence.Results:The HBV transgenic mice have stably bred to 23rd generation.The serum HBsAg level is 4122.31?2044.74IU/ml;The rate of HBV transgenic mice whose serum HBV DNA reach 104~106copies/ml was 93.93%.The HBV replication and expression were improved markedly.There is no difference between male and female mice about serum HBsAg level.Conclusion:After breeding the HBV gene was expressed stably with high-level in transgenic mice.

AIMInducing mesenchymal stem cells (MSCs) differentiate to myoblasts with 5-azacytidine(5-Aza-CR), investigating the expression of Myf5 and the role of the signal transduction case of p38 in all the course of differentiation.

METHODSSeparating and purifying bone marrow-derived MSCs, inducing MSCs differentiation to myoblasts with 10 micromol/L 5-Aza-CR, assaying the gene expression time of Myf5 with RT-PCR method, the antigen expression of myosin with immunohistochemistry method and observing the changes of the activity of phosphorylation p38 before and after inhibited by SB203580 with Western-blot method.

RESULTSMSCs begin to express Myf5 delayed to the 9th day after inhibited by SB203580. Some of MSCs express myosin at the 7th day after induced; The phosphorylation p38 activity of MSCs enhanced after induced by 5-Aza-CR but obviously decreased after inhibited by SB203580.

CONCLUSIONMSCs can express myogenic regulator factors and orientation differentiate to myoblasts after induced by 5-Aza-CR, p38 really have a positive signal transduction affection in this course.

Animals ; Azacitidine ; Bone Marrow Cells ; cytology ; Cell Differentiation ; Cells, Cultured ; Mesenchymal Stromal Cells ; cytology ; Mice ; Mice, Inbred BALB C ; Myoblasts ; cytology ; Myogenic Regulatory Factor 5 ; metabolism ; Signal Transduction ; p38 Mitogen-Activated Protein Kinases ; metabolism

Animals ; Cell Proliferation ; Chemokine CXCL12 ; metabolism ; Hepatocytes ; cytology ; metabolism ; Male ; Rats ; Rats, Wistar ; Receptors, CXCR4 ; metabolism

Sixty-one endophytic fungus strains with different colony morphologies were isolated from the leaves, stems and roots of Tephrosia purpurea with colonization rates of 66.95%, 37.50%, and 26.92%, respectively. Based on internal transcribed spacer sequence analysis, 61 isolates were classified into 16 genera belonging to 3 classes under the phylum Ascomycota. Of the 61 isolates, 6 (9.84%) exhibited antifungal activity against one or more indicator plant pathogenic fungi according to the dual culture test. Isolate TPL25 had the broadest antifungal spectrum of activity, and isolate TPL35 was active against 5 plant pathogenic fungi. Furthermore, culture filtrates of TPL25 and TPL35 exhibited greater than 80% growth inhibition against Sclerotinia sclerotiorum. We conclude that the endophytic fungal strains TPL25 and TPL35 are promising sources of bioactive compounds.
Ascomycota ; Colon ; Fungi* ; Plants ; Sequence Analysis ; Tephrosia*

Objective To develop a cellular shear stress loading system with an adjustable stress mode and relevant parameters, and subsequently verify the effectiveness and feasibility of this system. Methods The hardware of the system was developed by using a peristaltic pump and self-designed multi-channel flow chamber, and the mode control program of shear stress based on LabVIEW was designed to control the device via RS485 interfacing and Modbus protocol. Additionally, the relationship between the shear stress and system parameters was calibrated, and finite element analysis was also conducted. Finally, the feasibility of the system was confirmed via the in vitro cell experiment. Results The mode and magnitude of shear stress of the system could be controlled via either the peristaltic pump or computer, and the cellular long-term effect was also able to be detected. The calibration results of the system indicated that the level of shear stress exhibited significantly linear positive correlation with the revolution of the peristaltic pump (P<0.001). Finite element analysis demonstrated that the level of shear stress on the slide was uniformly distributed and the result of simulation was accordant with calibration. Cytoskeleton staining suggested that cellular morphology of MLO-Y4 cells was changed, and microfilament increased and arrayed along fluid flow direction. Conclusion The system is stable and reliable enough to provide different loading modes and magnitude of cellular shear stress to offer a convictive platform of the research for different cellular stress signal transduction mecha-nisms.

Objective To develop attenuated Salmonella which harboring enterovirus 71 (EV71) VPI gene. Methods The plasmid which expressed VP1 protein of EV71 was constructed by gene recombination. Cellular expression was assessed by Western Blot analysis. The recombinant plasmid was then transformed into attenuated Salmonella SL7207. Results EV71 VP1 gene sequence was inserted into a eukaryotic expression plasmid VR1012. VP1 protein was detected by Western Blot analysis in the culture supernatant. And the attenuated Salmonella harbored the plasmid stable. Conclusion The plasmid was constructed successfully and it can express effectively in vitro. The bacteria which harboring the plasmid were constructed successfully. This has provided a basis for further research of an oral EV71 vaccine.