BACKGROUND: Recent studies indicate that after indirect co-culture of neonate rat myocardial cells and bone marrow mesenchymal stem cells, bone marrow mesenchymal stem cells can differentiate into myocardial cells successfully. OBJECTIVE: To induce human bone marrow mesenchymal stem cells (hBMSCs) to differentiate into endothelial cells using human umbilical vein endothelial cells by indirect co-culture. DESIGN, TIME AND SETTING: In vitro study of cell engineering was done at the Medical Research Center of Guangdong Provincial People’s Hospital between January and July 2007. MATERIALS: Small quantities of bone marrow were obtained from 11 children with congenital heart disease but without hematologic diseases through manubrium of sternum puncture in the congenital heart defect corrective surgery after the permission of family member. Umbilical cord of full-term normal delivery healthy newborn was provided by the Department of Obstetrics, the First Affiliated Hospital of Sun Yat-sen University. Fresh cattle jugular vein was provided by Guangdong Dali Meat Cattle Butchery. METHODS: The hBMSCs were isolated and purified using density gradient centrifugation method and were cultured in vitro. Human umbilical vein endothelial cells were obtained from newborn umbilical cord by enzyme digestion. Cell culture insert with semipermeable membrane combined with 6-well plate was used to do indirect co-culture induction. Human umbilical vein endothelial cells were expanded in the cell culture insert, passage 3 bone marrow mesenchymal stem cells were expanded in the 6-well plate outside the culture insert at the density of 1?105 cells/well, the initial ratio of the two kinds of cells was 1:5, then low-glucose DMEM culture solution containing 10% fetal bovine serum was added, cells were cultured for 14 days. Co-culture of bone marrow mesenchymal stem cells and bone marrow mesenchymal stem cells was used as control. Introduced endothelioid cells were cultured and then seeded on the cell-free cattle jugular vein intravascular stent. MAIN OUTCOME MEASURES: Morphology changes of induced cells; introduced endothelioid cell surface antigen detected through immunocytochemical staining; the growth and adhesion condition of endothelioid cells on the intravascular stent observed under scanning electronic microsope. RESULTS: The morphologies of introduced endothelial cells were uniform, introduced endothelial cells presented a cobblestone-like appearance, they amplified fast and expressed endothelial cell-specific surface marker CD31 and vWF and the positive rate was over 99%. They also could form a continuous unicellular layer on the cell-free cattle jugular vein intravascular stent. CONCLUSION: Human umbilical vein endothelial cells can induce hBMSCs to differentiate into endothelial cells successfully and to adhere and grew on the cell-free cattle jugular vein intravascular stent through indirect co-culture method.

0.05), but the patients in the rehabilitation group demonstrated much higher scores in Fugl Meyer and Barthel index than those in the control group at the end of 4 and 12 weeks after rehabilitation therapy( P 0.05) 12 weeks after the therapy. Conclusion Early intensive rehabilitation therapy can effectively improve the motor function and ADL of stroke patients; the continuing home rehabilitation program for at least 8 weeks is helpful for improving the patient's functional independence, increasing the cost effectiveness, and shortening the length of stay of stroke patients.

Objective To investigate the optimal combination and the value of functional magnetic resonance imaging in the diagnosis of breast tumors. Methods One hundred and forty patients who had been clinically and pathologically diagnosed as breast tumors in our hospital during the period of 2009 to 2013 were collected. 63 of whom had breast cancer and 77 had benign tumor. All the patients received 1H-MRS, DWI and dynamic enhanced MRI at the same time. The obtained images were analyzed and then compared with the pathological findings. The accuracy, sensitivity, and specificity of MRS combined with DWI or dynamic enhanced MRI, or DwI combined with enhanced MRI in the diagnosis of breast cancer were obtained. Results All the diagnosis were pathologically confirmed. The accuracy, sensitivity, and specificity were 89.8%, 90.1%, and 90.0% for DWI combined with dynamic contrast-enhanced MRI; 77.9%, 77.3% and 79.3% for MRS in combination with enhanced MRI; and 76.3%, 77.8% and 77.1% for DWI combined with MRS. There were significant differences among the three kinds of combination detection in diagnosis of breast cancer , χ2= 9.057, P = 0.011. Conclusions The sensitivity, specificity and accuracy is the highest in DWI combined with dynamic enhanced MRI in the diagnosis of breast tumor , which can be used as the best combination imaging for breast tumor.

Objective To investigate the mid-term effect of SolitaireAB stent-assisted interventional embolization with spring coils for the treatment of intracranial wide-necked aneurysms. Methods During the period from May 2009 to April 2013, a total of 49 patients with intracranial wide-necked aneurysm (49 aneurysms in total) received SolitaireAB stent-assisted interventional embolization treatment at authors’ hospital. In 41 patients, a total of 41 aneurysms were detected, of which ruptured aneurysm with bleeding was confirmed in 26 and un-ruptured aneurysm in 15. These 41 patients were followed up for 12-48 months. Based on modified Rankin scoring and DSA, CTA or MRA manifestations, the clinical results were evaluated. Results After the embolization treatment, re-bleeding of the aneurysm occurred in 2 cases, cerebral infarction in 3 cases, occlusion of the parent artery in one case and death in one case; the occurrence rate of complications was 14.2%. DSA, MRA or CTA performed at 12 months after the embolization treatment, showed that 32 aneurysms (78.0%) were completely obstructed, which was obviously higher than that observed on DSA performed immediately after the embolization procedure (21 aneurysms, 42.9%), the difference was statistically significant (P=0.02);residue of aneurismal neck was seen in 7 cases (17.1%) and partial occlusion in 2 cases (4.9%), which were much better than those observed on DSA that was performed immediately after the embolization procedure. Twenty-four aneurysms (58.5%) remained stable, showing no any change, and recurrence of aneurysm was observed in 4 cases (9.7%). At the last follow-up exam, the modified Rankin scoring showed that 0 point was seen in 18 cases (43.9%), one point in 10 cases (24.4%), 2 points in 5 cases (12.2%), 3 points in 4 cases (9.8%), 4 points in 2 cases (4.85%) and 5 points in 2 cases (4.85%). The self-care rate for daily activities was 80.5%, the prognosis was good. Conclusion For the treatment of intracranial wide-necked aneurysms, SolitaireAB stent-assisted interventional embolization with spring coils is safe and effective. This technique can improve the embolization rate and reduce the procedure-related complications.

Dao-di Herbs is specificity and locality, and its unique phenotypic features is closely related to the growth and development of medicinal plants. In addition to traditional genetic, epigenetic play an important role in formation of Dao-di herbs. This paper introduces the concept of epigenetic and the role of DNA methylation in the gene expression regulation. We further prospects epigenetic mechanism in study of Dao-di herbs formation from specific phenotype and regional analysis. And study on the relationship of epigenetic and Dao-di herbs will provide a basis for quality assessment and identification of Chinese drugs.
DNA Methylation ; Drugs, Chinese Herbal ; standards ; Epigenesis, Genetic ; Gene Expression Regulation, Plant ; Plants, Medicinal ; genetics

In the research field of quality control in Chinese medicinal materials, variation in active ingredients of medicinal plant is always the key and hot issues. With the development of high-throughput sequencing technologies and reducing cost, a large numbers of genes from medicinal plant were cloning and provide a solid foundation for further research of gene structure and its biological function, and also provides conditions for explore active ingredient variation and its quality control from the perspective of molecular pharmacognosy. This paper introduces the concept of homologous gene, gene duplication and classification. We prospect the function of duplicated genes in the role of molecular mechanism research about variation in active ingredients, aiming at providing a new way for medicinal materials quality control.
Drugs, Chinese Herbal ; analysis ; Gene Duplication ; Plant Proteins ; genetics ; Plants, Medicinal ; chemistry ; genetics ; Quality Control

BACKGROUND:It is shown in past researches that total saponin of ginseng has the effect of promoting multiplication of bone marrow hematopoietic stem cells and progenitor cells in vitro, and promoting secretion of bone marrow hemotopoietic factors, however there has been little researches on the effect of multiplication of bone marrow stromal cells.OBJECTIVE: To investigate the effect of ginsenoside Rg1 on pig's bone marrow stromal cells multiplication in vitro.DESIGN: A controlled observation.SETTING:Department of Cardiology and Department of Dermotology,First Affiliated Hospital, Nanjing Medical University; Department of Pharmacology, Nanjing Medical University.MATERIALS: The experiment was conducted at the Cardiovascular Pharmaceutical Laboratory of Nanjing Medical University from January 2002 to February 2003. The experimental pigs ahd the powder of ginsenoside Rg1were chosen METHODS :The bone marrow stromal cells cultured in vitro were divided as control and experimental groups. For experimental group insenoside Rg1 in different concentrations (10-7,5×10-7,10-6,5×104 mol/L)were added for induction culture, and for control group the media of the same volume was added. The multiplication conditions of bone marrow stromal cells were respectively detected by fibroblast colony forming in vitro, tritiated thymidine {[3H]TdR} incorporation and MTT.MAIN OUTCOME MEASURES: ① The effect of ginsenoside Rgl on the fibroblast colony forming units of bone marrow stromal cells in vitro. ②The effect of ginsenoside Rg1 on [3H]TdR incorporation of bone marrow stromal cells. ③ The effect of ginsenoside Rg1 on bone marrow stromal cells multiplication detected by MTT.RESULTS:① The effect of ginsenoside Rg1 on the fibroblast colony forming units of bone marrow stromal cells in vitro: The numbers of units in experimental group were all higher than that in control group (P ＜ 0.05-P ＜ 0.01),especially obvious in (5-50)×10-7 mol/L experimental group, reaching peak in the concentration being 10-6 mol/L. ② The effect of ginseno side Rg1 on [3H]TdR incorporation of bone marrow stromalcells: As the concentration of ginsenoside Rg1 was 10-6 mol/L, the [3H]TdR incorporation efficiency of bone marrow stromal cells was obviously higher that that in control group (P ＜ 0.05-P ＜ 0.01), and the effect was enhanced as the dosage increased. ③ The effect of ginsenoside Rgl on bone marrow stromal cells multiplication detected by MTT: The value of absorbance in experimental group was obviously higher than that in control group (P ＜ 0.05-P ＜ 0.01), and it was dose-dependent. CONCLUSION: Ginsenoside Rg1 had an effect of multiplication on pig's bone marrow stromal cells in vitro, and it was dose-dependent.

Objective To determine whether the supplementation of thyroid hormone(TH)improves β-adrenoceptors(β-AR)density and expression of peripheral blood lymphocyte in elderly patients with chronic heart failure(CHF). Methods A total of 42 elderly patients with advanced CHF(NYHA classⅢorⅣ)due to dilated cardiomyopathy(DCM)or ischemic cardiomyopathy (ICM)were randomly divided into a treatment group and a control group.Low-dose L-thyroxine(LT50)was added to the treatment group,both groups had routine anti-heart-failure treatment.Before and after 1 month of treatment,125I-Pindolol radioligand binding assay was used to measure β-AR in peripheral lymphocytes.Reverse-transcriptase polymerase chain reaction(RT-PCR)was used to measure β1-AR expression in peripheral lymphocytes. Results The density and expression of β-AR of lymphocytes in patients with CHF due to ICM or DCM were lower than those in normal health people.and the serum TH level in patients with CHF was lower than that in normal health people.The L-T4 administration induced an up-regulation of β-AR density and expression in peripheral lymphocytes. Conclusions Thyroxine administration induces an up-regulation of β-AR density and expression in peripharal lymphocytes.

BACKGROUND: Extracorporeal shock wave therapy is indicated as an effective method for treatment of delayed fracture healing or nonunion. Osteoblasts plays an important role in this process.OBJECTIVE: To investigate the function of osteoblasts in the process of extracorporeal shock wave promoting fractures healing, and to provide theoretical support for improving shock wave therapy on fracture healing.METHODS: Primary cultured ostsoblasts were isolated from newbon SD rat calvaria and randomly divided into two groups, shock wave and control. Treated by different energies of extracorporeal shock wave, cells were incubated onto 96-well culture plate. An optimal dose of extracorporeai shock wave was selected according to survival and proliferation of osteoblasts. The osteoblasts treated by optimal energy of extracorporeal shock wave were cultured and harvested for the analysis of alkaline phosphatase by calcium cobolt stain, cell survival by CCK-8 Kit, alkaline phosphatase expression by AKP kit, mineralized nodules by Alizarin red staining, integrin β1 and β1 mRNA expressions by flow cytometry and RT-PCR, cell migration by wound healing assay.RESULTS AND CONCLUSION: The optimal energy of extracorporeal shock wave treating primary cultured osteoblasts was 10 kV (500 impulses). Following extracorporeal shock wave therapy, the cell proliferation, alkaline phosphatase activity, cell mineralization, rates of cell adhesion, as well as β1 integrin and its mRNA expressions were increased as compared with those in control group (P ＜ 0.01). Further distance of cell migration was found in extracorporeal shock wave group (P ＜ 0.05). The results showed that the optimal energy of extracorporeal shock wave could promote the proliferation, differentiation, adhesion and migration of osteoblasts in vitro, and β1 integrin may play an important role in the process of cell adhesion and migration.

AIM:To study the chemical constituents of the leaves of Cyclocarya Paliums(Batal.) Iljinskaja. METHODS: Chemical constituents were separated through chromatographic methods and their structures were elucidated by spectral analysis. RESULTS: Six compounds were isolated from the chloroform fraction through 75% alcohol extraction from the leaves of Cyclocarya Paliums.Their structures were elucidated as ?-Sitosterol(1),Behenic acid(2),3?,20-dihydroxyl-5?-Pregnanes(3),Kaempferol(4),?-amyrin(5),?-amyrone(6). CONCLUSION: Compounds 3?,20-Dihydroxyl-5?-Pregnanes(3),?-amyrin(5) and ?-amyrone(6) are isolated from this plant for the first time.