[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 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 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.

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.

Objectives To investigate the pathophysiologic mechanisms and the clinical character- istics and variations of nutcracker esophagus(NE).Methods Clinical data obtained from 22 patients with NE were retrospectively analyzed.Seven followed-up patients had esophageal motility,multi-channel electrogastrography(MEGG),the autonomic nervous system(ANS) and psychology tests and were com- pared to 10 healthy subjects(HS).Results①In NE group,12 patients had reflux symptom(55%),7 patients had chest pain(32%)and 3 patients had dysphagia(13%).There was no statistical difference in mean contraction amplitude(MCA) between patients with reflux symptom and chest pain.②Eight out of the 13 patients with NE who received 24 h pH monitoring were positive reflux,and 4 out of 17 patients had reflux esophagitis in endoscopic examinations.The symptoms were improved in 58%patients(7/12) by regular acid-suppression therapies.③There was no statistical difference beween NE and HS groups in dominant frequency and power of MEGG.However,the percentages of normal rhythm in preprandial and slow wave coupling in pre and postprandial of NE patients were significantly decreased than those in HS.④The ANS function in NE group had no statistical difference compared to those in HS.⑤In follow- up group,no difference was found before and after nitroglycerin sublingually.Four patients had depres- sion.Conclusions The clinical presentations of NE are vary.The symptoms of NE were poorly correla- ted with MCA,but partially correlated with GER,which may represent a special subtype of GERD. Gastric dysmotility and psychological factors might contribute to the pathogenesis of NE.

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.