OBJECTIVETo investigate the effects of E 4031, a sodium calcium exchanger (NCX) agonist, on the isolated cardiac function and resting Ca2+ level in myocardial cells from rats with chronic heart failure.

METHODSRats chronic heart failure model was established by abdominal aorta coarctation with; Isolated heart perfusion by Langendorff apparatus was used to detect heart function and the effects of E 4031 on haemodynamic indexes; Myocardial cells of rats in the model group were extracted quickly and co-incubated with calcium fluorescent indicator fluo3/AM and the impact of E 4031 on the fluorescence intensity in myocardial cells were evaluated by confocal microscopy.

RESULTSHeart function of rats in the model group detected by Langendorff perfusion was significantly reduced after 12 weeks, E 4031 at 10 micromol/L could improve their left ventricular developed pressure(LVDP) and systolic / diastolic maximum rate (+/- dp/dtmax). Compared with the control and sham operation groups, the resting Ca2+ fluorescence intensity of the myocardial cells of rats in model group was at a higher level and went through a process of transient rise and drop, then stably remaining at a low level after co-incubated with 10 micromol/L E 4031.

CONCLUSIONE 4031 can improve the isolated heart function of rats with chronic heart failure, which may be associated with its enhancing the activity of NCX in the myocardial cell membrane and stabilizing intracellular Ca2+ level.

Animals ; Calcium ; metabolism ; Disease Models, Animal ; Heart ; physiopathology ; Heart Failure ; metabolism ; physiopathology ; In Vitro Techniques ; Male ; Myocardium ; metabolism ; Myocytes, Cardiac ; metabolism ; Piperidines ; pharmacology ; Pyridines ; pharmacology ; Rats ; Rats, Wistar

Objective To design and construct the vector of protein-rich tyrosine kinase 2 small RNA interference and investigate the effect of the recombinant plasmid on the proliferation of adult rat cardiac fibroblasts. Methods The pAVU6+27-PYK_2 siRNA expression vector was constructed by gene recombination, then transfected into the cultured adult rat cardiac fibroblasts by DOTAP method. PYK_2 mRNA and protein in cardiac fibroblasts were determined by RT-PCR and Western blotting respectively. Cell proliferation was tested by MTT and ~3H-TdR incorporation. Results The pAVU6+27-PYK_2 siRNA expression vector was successfully constructed. The recombinant plasmid can inhibit the proliferation of adult rat cardiac fibroblasts, as compared with control. Conclusion pAVU6+27-PYK_2 siRNA expression vector is efficient to suppress DNA synthesis of cardiac fibroblasts, which will be beneficial to further study of myocardial fibrosis.

Objective To explore the potential of breast cancer stem cells in differentiating into vascular endothelial cells and participating in the angiogenesis of breast tumor.Methods The expressions of mutant P53, CD31 ,VEGF and Her-2 amplification in tumor blood vessels were detected and the single cell suspension of breast cancer was prepared.CD44+/CD24-/low cells were selected and cultured in culture system.The expressions of CD31 and CD105 on endothelial cell surface were detected by EGM-2 endothelial cell culture medium.The ability of uptake of acetylated low-density lipoprotein was measured.The endothelial cells were cultured in vitro to observe their vascular structure.Results The expressions of CD31,VEGF and mutant P53 were found in paraffin sections of breast cancer tissue samples, and they were arranged along the vessel lumen or blood vessel sphere. Immunofluorescence showed that CD31 and DAPI signals were expressed intravascularly in breast cancer tissues. Four samples of Her-2 positive amplification and 9 cases of non-amplification were detected in the samples.The successful separation of breast cancer stem cells was carried out by immunomagnetic sorting.The percentage of CD44+ cells in the pre-sorting cells was (7.5±2.6)% and that of CD44+ cells was about (94.3±4.7)% after magnetic sorting (P<0.05).The ratio of CD24+ cells was (48.2±9.4)% before sorting,and that of CD24+ cells was (4.3±1.6)% after immunomagnetic bead sorting.The proportion of CD105+ cells in mammary gland cells was (4.5±0.9)% and the proportion of CD31+ cells was (6.2±1.3)%.After cultured for three generations,the percentages of CD105+ cells and CD31+ cells were (79.6±9.3)% and (84.1±10.7)%,respectively (P<0.05). DiL-Ac-LDL could be phagocytized by endothelial cells cultured in endothelial cell culture system.Vessel-like structure was found in the endothelial cell group while the control group did not have the tendency of vascular changes.Conclusion Breast cancer stem cell-derived endothelial cells differentiate into endothelial cells in vitro and have the ability of vascular formation,suggesting that breast cancer stem cells may be involved in the formation of tumor blood vessels.

Adenoma ; Adult ; Ear Neoplasms ; Ear, Middle ; pathology ; Humans ; Male

Objective To investigate effects of different doses of recombinant human bone morphogenetic protein-2 (rhBMP-2) on vascular endothelial growth factor (VEGF) expression in fetal mouse osteoblasts.Methods Calvaria osteoblasts of fetal mice of 19 days were cultured.The effects of rhBMP-2 at different doses and different action times on VEGF expression patterns in fetal mouse osteoblasts were observed with reverse transcrip- tion-polymerase chain reaction (RT-PCR) and Western blot staining.Results In the present study,RT-PCR detected a steady expression of VEGF mRNA in the control fetal mouse osteoblasts.The levels of VEGF mRNA increased in an apparent biphasic manner with a maximum stimulation (about 2-fold above the control,P＜0.05 ) in both VEGF mRNA species observed at 300 ng/mL of rbBMP-2.After 48 h of rhBMP-2 treatment,the VEGF mRNA levels approached those in the control.The VEGF mRNA levels appeared to be biphasic in rhBMP-2-treated cultures,showing peak induction at 3 and 24 h and remaining elevated at 48 b.Compared with the individual control value at each time point,an apparent maximum increase (about 2.5-fold above the control,P＜0.05) occurred at 6 h.The second peak induction,about 2-fold above that in the control,occurred at about 36 h.Conclusions The expression of VEGF mRNA is steady in the control fetal mouse osteoblasts.RhBMP-2 can promote the expression of VEGF in dose-dependent and time-dependent manners.

OBJECTIVETo study the effects of dopamin receptors-2 (DR2) on myocardial ischemic postconditioning and explore its underlying mechanisms.

METHODSThe myocardial ischemic postconditioning (PC) model was established in cultured primary rat neonatal cardiomyocytes which were then randomly assigned in the following groups: Nomial control group, Isehemia/reperfusion (L'R) group, PC (ischemic postconditioning) group, PC + Bro (Bromocriptine, a DB2 antagonist) group, PC + Hal (Haloperidol, a DB2 repressor) and PC + Hal + Bro groups. The lactate dehydrogenase (LDH) and superoxide dismutase (SOD) activity and malondialdehyde (MDA) content in cell medium were analyzed by colorunetry. The cell ultrastructure changes were observed by transmission electron microscope. The cell apoptosis was analyzed using flowcytometiy. The protein expression level of D112 and activity of p-p38 and p-JNK were detected by Western blot.

RESULTSCompared with the nonnal control group, hR increased the protein expression level of DB2, enhanced LDH activity and MDA content, promoted cell injury and apoptosis, decreased SOD activity, up-regulated the activity of p-p38 and p-JNK. Compared with the hR group, although PC further increased the expression of DR2 protein, it decreased LDH activity and MDA content, cell injury and apoptosis, increased SOD activity, down-regulated activity of p-p38 and p-JNK. Bromocriptine treatment further enhanced PC-induced canlioprotective effect, yet Hal addition attenuated this enhancing effect exerted by bromocriptine.

CONCLUSIONThe activation of DB2 is involved in the protective effect of ischemic postconditioning on myocardial ischemia/reperfusion injury through down-regulating the activity of p-p38 and p-JNK.

Animals ; Apoptosis ; Cells, Cultured ; Ischemic Postconditioning ; JNK Mitogen-Activated Protein Kinases ; metabolism ; Myocardial Reperfusion Injury ; prevention & control ; Myocytes, Cardiac ; pathology ; Rats ; Rats, Wistar ; Receptors, Dopamine D2 ; physiology ; p38 Mitogen-Activated Protein Kinases ; metabolism

To understand the differences of the clinical research of acupuncture at home and abroad in methodological design and implementation, so as to provide reference for exploring acupuncture clinical study design program suitable to China. Reading clinical research literature of acupuncture and moxibustion for treatment of stroke at home and abroad and analyze and probe the methodological design and implementation. The results indicate that clinical research of acupuncture and moxibustion abroad has advantages in the randomized, controlled and double blind, etc. We should adopt the advantages of Chinese medicine, enrich and perfect research design program which can be approved by modern medicine and properly reflect the therapeutic effect of acupuncture and moxibustion on stroke.
Acupuncture Therapy ; Humans ; Medicine, Chinese Traditional ; Moxibustion ; Randomized Controlled Trials as Topic ; Research Design ; Stroke ; therapy

AIM:To explore a novel method to isolate human nucleus pulposus mesenchymal stem cells (hNP-MSCs) in vitro and to identify their biological characteristics.METHODS:The explant culture method was employed to isolate hNP-MSCs from nucleus pulposus tissue obtained by percutaneous endoscopic lumbar discectomy (PELD).The isolated cells were passaged for purification and cultured in vitro followed by morphological observation.The cell proliferation ability was detected by CCK-8 assay.Growth curves of the cells were drawn and surface antigens were detected by flow cytometry.The cells at the 3rd~6th passages were induced for adipogenic, osteogenic and chondrogenic differentiation, and examined by oil red O staining, alizarin red staining and Alcian blue staining.RESULTS:The cells with self-renewal were obtained from nucleus pulposus tissue obtained by PELD.The results of flow cytometry analysis revealed that the cells were positive for CD29, CD44, CD90, CD73 and CD105, but negative for CD34 and CD45.The proliferative capacity was consistent with the growth characteristics of MSCs and multilineage differentiation potential was identified.CONCLUSION:A novel method to efficiently isolate and culture hNP-MSCs,PELD combined with explant culture method,was established, which would promote the study of regenerative medicine based on hNP-MSCs.

Objective To evaluate the efficacy and safety of 308-nm excimer light in the treatment of stable vitiligo. Methods Thirty patients with stable vitiligo were enrolled in this clinical trial. All the subjects received the treatment with 308-nm excimer light on a twice-weekly schedule for 3 months. Results The repigmentation rate was 95.0%, 75.0% and 66.7% for lesions in the face and neck, trunk and limbs, with the treatment sessions averaging 10.22 ± 1.60, 19.10 ± 2.38, 37.74 ± 3.06, respectively, and accumulative irradiation dose averaging 7.50 ± 3.45, 10.60 ± 1.01, 18.56 ± 3.05 J/cm2 respectively. Significant differences were observed in the repigmentation rate and treatment sessions between the lesions in the face and neck, trunk and limbs (all P ＜ 0.05). No severe side effects were seen during the treatment. Conclusion 308-nm excimer light is effective and safe for the treatment of vitiligo.

Objective To evaluate the influence of laparoscopic uterine artery ligation on ovarian function. Methods In this retrospective study ,46 patients with laparoscopic myomectomy were selected and randomly divided into Ligation group and Non-Ligation group. The serum concentrations of follicular stimulating hormone (FSH), luteinizing hormone ( LH), estrogen ( E_2) were measured be-fore treatment and 1 ,3 ,6,12 months after treatment. Ovulating functions of ovary were monitored. All results were compared between two groups. Results All patients ovulated after 6 months. There were no significant differences between two groups in the levels of FSH, LH and E_2,.before and after treatment(P>0.05). Conclusions Laparoscopic uterine artery ligation do not affect ovarian function of pa-tients with uterine leiomyoma.