[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

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.

Objective To analyze the clinical data of recipients over 15 years after renal transplantation,and to find the factors that affect the long-term survival of recipients after renal transplantation.Method Before June 30,2000,326 renal transplant recipients in our hospital were collected retrospectively.The risk factors which affect the survival of kidney transplant recipients and kidney were analyzed from four dimensions.A Cox model was established to analyze these multi factors.Result Cox hazard model indicated that advanced age (P=0.010,RR =1.052),AMR (P＜0.001,RR =18.311),nonadherence (P =0.001,RR =2.854),smoking (P =0.025,RR =2.097)were the risk factors for recipients' survival.Using immunosuppressive regimen FK506 + MMF+ Pred (P =0.019,RR =0.433),or CsA + MMF + Pred (P =0.019,RR =0.413) was the protective factor for recipients' survival.Nonadherence (P＜0.001,RR =5.645),and diabetes (P＜0.001,RR =3.310) were the risk factors of grafts' survival.Using immunosuppressive regimen FK506 + MMF + Pred (P＜0.001,RR =0.236),or CsA + MMF + Pred (P =0.002,RR =0.317) was the protective factor of grafts' survival.Conclusion To enhance the long-term outcome of recipients and grafts,the individualization of immunosuppressive regiments and controlling of the chronic diseases progress by changing the unhealthy life style are cutting on edge.

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.

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.

Objective To explore the effects of coincident infection on treatment response and coronary artery lesion (CAL) outcome in children with Kawasaki disease(KD).Methods A retrospective study of 141 children diagnosed on KD between Jul.2005 and Dec.2006 were performed.Standardized clinical assessments,laboratory examinations microbiology test results plus treatment regimens were reviewed.CAL were visualized by using echocardiography.Infectious agents positive (INF+) and negative (INF-) groups were identified,and clinical assessments,laboratory and treatment data were analyzed.Results 1.Concurrent infections:41%(58/141) of children had one of above confirmed infection at KD diagnosis.2.Treatment response:the presence of infection did not alter the response to treatment with intravenous immunoglobulin (IVIG),with resolution of fever within 72 h in 85% (120/141) children after 1 dose of IVIG (2 g/kg) together with aspirin administration regardless of present or absent infection.3.CAL outcome:in total,56.0% (79/141) of children developed CAL at the time of diagnosis,involving dilatation (91.1%,72/79 cases) and aneurysm (8.9%,7/79 cases),and no giant aneurysm was found.Most CAL were recovered within 1 year after treatment.Incidence of aneurysm in INF+ group was significantly higher than that in INF-group (P=0.019).Conclusions Coincident infection would not affect on the clinical assessment,laboratory test results and treatment response to IVIG in children with KD,but would result in higher risk of serious CAL.Therefore,children with infection at diagnosis on KD will not only accept active treatment in acute phase,but also insist on convalescent care and follow-up visit.

Objective To investigate relationship between levels of follicle-sitimulating horomone receptor(FSHR) and ovarian response induced by gonadotropin hormone,and whether the FSHR expression is correlated with in vitro fertilization(IVF) outcome.Methods Granulose cells were collected from 43 women receiving IVF-embryo transplantation(IVF-ET).According to the number of oocyte,the women were divided into three groups: low response(15).The expression intensity of FSHR was measured by immunohistochemistry technique.The expression intensity of FSHR on the granular cell,the embryological and clinical outcomes were compared and analyzed. Results The expression of FSHR was significantly different in three groups with the highest in high response group(P0.05).The FSHR level was positively correlated either with the number of oocyte (r=0.719) or with the serum E2 levels(r=0.516,P0.05). Conclusion Ovarian response to gonadotropin hormone stimulation is correlated with the level of FSHR in the granulose cells.The development of follicles may be influenced by it.