[Objective] To establish a method to seperate rat mesenchymal stem cells(rMSCs) from rat marrow, culture rMSCs in vitro, and study its biological characteristics. [Methods] rMSCs were seperated from rat marrow with wall-sticking method and expanding cultured in vitro. To draw the growth curves of cells at 1th, 3th, 5th passage, detect the surface antigens with flow cytometry. To evaluate the effect of fetal calf serum with different concentrations to rMSCs. [Results] There were approximately (5～6)?105 cells after primary culture and over (2~3)?108 rMSCs at 5th passage. The appearance of rMSCs is like shape of spear and its growth curves are like shape of S. The rMSCs were positive for CD90 and negative for CD45. [Conclusions] Cells seperated and cultured by the method established in this experiment were rMSCs. They have the biological characteristics of rMSCs, and are good resources for study in tissue engineering.

Administration, Rectal ; Adolescent ; Adult ; Drugs, Chinese Herbal ; administration & dosage ; Endometriosis ; drug therapy ; Female ; Humans ; Middle Aged ; Phytotherapy

Animals ; Intestines ; cytology ; drug effects ; physiology ; Meperidine ; pharmacology ; Mice ; Mice, Inbred Strains ; Muscle, Smooth ; drug effects ; physiology ; Rabbits

The reproduction of Helicoverpa armigera nucleopolyhedrovirus in the midgut epithelia cells and the other sensitive tissues was observed by electron microscopy. The reproducing viruses in the midgut epithelia cells were mostly without envelopes, and thte polyhedrons were seldom formed. The reproduciing viruses in the other sensitive cells were with envelopes, and packed in polyhedrons.

At the beginning of the 1980s, a concept of viable but non-culturable(VBNC) was suggested. VBNC is a survival strategy adopted by microorganisms when they are exposed to environmental stress. This article try to make a summary of research of the conditions of VBNC formation, recovery of culturability and methods of VBNC cells detection. In addition, introduces the first growth factor of microorganisms-Rpf.

Objective To probe the value of using echocardiography in transcatheter closure of ventricular septal defect (VSD).Methods Under transthoracic echocardiography and digital subtraction angiocardiography monitoring and guidance in the operation,the Amplatzer excentric umbrella occluder was used to occlude the perimembranous defects in 15 cases of patients.Results All of the patients were treated successfully,though mitral regurgitations or residual shunts occurred in 6 cases,and disappeared at once after the position and direction of occluder were regulated in 3 and these complications disappeared gradually one month postoperation in other 3 patients.The umbrella separated from VSD in 1 and occluded successfully when manipulated once again.Complete heart block occurred in 2 and disappeared after 3 days.The positions of umbrella were normal and stable and no other complications were discovered when these patients were followed about 10 months the longest.Conclusions Echocardiography plays an important role in transcatheter closure of VSD with the Amplatzer occluder before,during and after operation and in the period of follow up.

Objective To investigate the correlation between pathogens and spectrum of disease in infants with human cytomegalovirus(HCMV) active infection.Methods A total of 378 cases of HCMV infection diagnosed by the identification of HCMV IgM or PP65 antigen of HCMV.HCMV gB genotyping was carried out by nested PCR and restriction fragment length polymorphism(RFLP) in 107 cases.The results of pathogen,spectrum of disease and clinic feature were analyzed.Results In all 378 infant patients with HCMV,27.78% were systemic infection and 72.22% involved just single organ.Hepatitis,HCMV inclusion disease,thrombocytopenic purpura,pneumonia were pre- dominant with 33.07%,27.78%,13.49%,6.35% respectively.The rate of HCMV inclusion dis ease in infants younger than 2 weeks was higher than in those aged from 3 12 weeks(P ~ 0.05) and children older than 12 weeks(P＜0.01).Infants with higher rate of PP65 antigen positive cells were apt to systemic infection than those with lower rate of PP65 positive cells(P＜0.01).Infants,who were positive by detections of all three methods,were apt to systemic infection than others(P＜0.01). Moreover,infants positive of IgM and PP65 antigen were apt to systemic infection than those just positive by one of the two methods(P＜0.01).The result of gB genotype analysis in 107 cases showed 53 cases of gBⅠ,20 of gBⅡ.18 of gBⅢ.7 of gBⅠ+gBⅡ,5 of gBⅠ+gBⅢand 4 of gBⅡ+gBⅢ,and gBⅣwas not found.Conclusion HCMV could infect multiple organs and have some different clinic features.Combination of different methods can increase the sensitivity to detect the pathogen.The gBⅠgenotype is most prevalent in these infants.

Objective To investigate effects of Kanglaite injection on proliferation, cycle and apoptosis of human breast cancer MCF-7 cells;To discuss its relevant mechanism. Methods Logarithmic growth phase cells were divided into control group and Kanglaite-treatment group (10, 20, 40μL/mL). Cells were cultured in RPMI-1640 for 24 h before drug treatment. The inhibition rate of Kanglaite injection on proliferation of human breast cancer MCF-7 cells was detected by MTT assay. Apoptosis and cell cycle of MCF-7 cells were detected by flow cytometry. Changes in cell nucleus were determined by Hochest staining assay. Protein expressions of Bcl-2 and Bax were detected by ELISA and Western blot. Results Kanglaite injection for 12 h, 24 h or 48 h resulted in a significant inhibition of MCF-7 cells proliferation (P<0.05, P<0.01);Compared with the control group, Kanglaite injection-treated cells showed increased percentage in G2/M and G0/G1 phases (P<0.001, P<0.01), but showed decreased percentage in S phase (P<0.01), and apoptosis rate increased (P<0.05, P<0.001). Kanglaite injection significantly decreased protein expression of Bcl-2, and enhanced protein expression of Bax of MCF-7 cells (P<0.01, P<0.001). Conclusion Kanglaite injection can inhibit the proliferation of human breast cancer MCF-7 cells, decrease cell cycle and induce apoptosis, the mechanism is related with decreasing protein expression of Bcl-2 and enhance the protein expression of Bax.

Objective To establish a new effective method by using real-time polymerase chain reaction (PCR) to detect single nucleotide polymorphism (SNP) typing for rapid identifying apolipoprotein E alleles.Methods To determine alleles of human apolipoprotein E genetic polymorphism at Cys112Arg locus was detected by PCR melting curve analysis with fluorophore SYBR Green I. In order to increase the speciality of SNP assays, high fidelity Taq polymerase was used. The reliability of SNP typing was validated by comparison with the results of direct DNA sequencing.Results Each sample was determined by double tubes, and two melting curves were analysis. As compared the Tm value of samples with the Tm of standard substance, the apoE genotype of samples was determined. The apoE genotype of 30 samples were E3/3 (27/30) and E3/4 (3/30) respectively, which was accordant with the results of PCR-RFLP and DNA sequencing.Conclusion The presented allelic assay was specific, easy to operate and applicable for discrimination of apolipoprotein E genotyping of human blood.

Long-term treatment with an agonist of peroxisome proliferator-activated receptor (PPAR)-γ is associated with bone fractures in the clinical practice. However, the mechanisms underlying the fractures are not fully understood. This study was aimed to examine the effect of rosiglitazone (an agonist of PPAR-γ) of different doses on the proliferation, differentiation, and transforming growth factor beta 1 (TGF-β1)-induced expression of connective tissue growth factor (CTGF) in primary rat osteoblasts in vitro. Osteoblasts were isolated from newly born SD rats and treated with different doses of rosiglitazone (0-20 μmol/L). The proliferation and differentiation of osteoblasts were measured by MTT assay and NPP assay, respectively. The expression of CTGF was determined by RT-PCR and Western blotting. The results showed that most isolated osteoblasts displayed strong alkaline phosphatase (ALP) activity and treatment with different doses of rosiglitazone did not affect their proliferation, but significantly inhibited the differentiation of osteoblasts in a dose-dependent manner. Moreover, treatment with different doses of rosiglitazone significantly reduced the TGF-β1-induced CTGF mRNA transcription and protein expression in a dose-dependent manner in rat osteoblasts. It was concluded that the activation of PPAR-γ may inhibit the differentiation of osteoblasts by reducing the TGF-β1-induced CTGF expression in vitro.