BACKGROUND: In vitro cultured bone marrow mesenchymal stem cells (BMSCs) can differentiate into neural cells. Some inductors are poisonous and cannot be used in human. OBJECTIVE: To induce the differentiation of rat BMSCs into neuron-like cells with astragalus mongholiens. DESIGN, TIME AND SETTING: The randomized control experiment was performed at the Yunnan Key Laboratory of Faculty of Basic Medicine of Dali University and Stem Cell Research Center of College of Preclinical Medicine and Forensic.Medicine of Sichuan University from January 2002 to January 2005. MATERIALS: Healthy SD rats aged 6-weeks old and weighing 120-130 g were used in this study. METHODS: BMSCs were collected from rat bone marrow by density gradient centrifugation, and then cultured. BMSCs were incubated in serum-free L-DMEM medium containing 0.125 volume fraction of astragalus mongholicus to observe morphologic changes in differentiated cells. Expression of the nestin, neuron specific enolase (NSE) and glial fibrillary acidic protein (GFAP) was detected by immunohistochemistry. MAIN OUTCOME MEASURES: Immunohistochemistry of differentiated cells; The expression of Wnt-1 gene and Ngn-1 gene were detected by semi-quantitative reverse transcdption-polymerase chain reaction during cell differentiation. RESULTS: After passage, BMSCs were fibroblast-like, and had high reproductive activity. After cultured with astragalus mongholicus, BMSC morphous changed. Nestin, NSE and GFAP were positive. The expression of Wnt-1 gene and Ngn-1 gene increased after astragalus mongholicus treatment. CONCLUSION: BMSCs can differentiate into neuron-like cells induced by astragalus mongholicus. Wnt-1 gene and Ngn-1 gene play a positive regulatory effect during the differentiation.

OBJECTIVE:To approach the correlations of carbamazepine concentration in serum with clinical therapeutic effect,dosage,sex,age and body weight in epileptic children.METHODS:158 monitoring records for serum concentration of carbamazepine in children with epilepsy in our clinical pharmacy department were consulted retrospectively.Data such as serum concentration,sex and age were recorded into data base and were analyzed using SPSS 13.0 software.RESULTS:No significant correlation was noted between serum concentration of carbamazepine in epileptic children and sex.However,serum concentration of carbamazepine was closely correlated with clinical therapeutic effect,dosage,age and body weight.CONCLUSION:The individual difference in the metabolic processes of carbamazepine was concerned with age and body weight,so it's necessary to make individualized regime according to child's specific condition and monitoring results.

To observe the changes of artrial natriuretic peptide (ANP) and endothelin (ET) lev-els in patients with c0ngenital heart disease (CHD) and discuss their mechanism and clinical significance.Methods: The pulmonary pressure of the pulmonary hypertension (PH)group was higher than 2 kPa- Thepulmonary pressure of Non-PH group was n0rmal. Healthy children served as control group. Results: (1)Plasma ANP level of the PH group was apparently higher than that 0f the N0n-PH group and c0ntrolgroup. (2) Plasma ET levels in the peripheral vein, vena cava,right atrium, right ventricle, pulmonaryartery and left atrium of the PH group were particularly higher than that of the Non-PH group and c0ntr0lgroup, especially in the left atrium. C0nclusi0n: (l) CHD with PH induces the increase in cardiac v0lumeand pressure load, thus accelerates the release 0f ANP. It is actually a compensatory mechanism for the el-evation of endogenic ANP. (2) ET is a vasoconstrictor. ET owns the importance at PH pathogenesis andhas a certain clinical value in the diagn0sis 0f PH. (3) ET causes ANP releasing, then inhibits ET effectthrough negative feed. B0th of them are antag0nists each other.

Objective To investigate potential protection by protect against ethanol-induced apoptosis of spermatogenic cells in rat testis.Methods Thirty SD adult male rats were randomly divided into three groups: normal group,alcohol group and puerarin group.At 40th day,BCL-2 and BAX of spermatogenic cells of testis tissue were checked by RT-PCR and immunohistochemistry;Apoptosis of spermatogenic cells was determined by TUNEL.Results The results of RT-PCR and immunohistochemistry indicated that BCL-2 and BAX of spermatogenic cells were not significanty different between puerarin group and normal group,but there was the significant difference between alcohol group and puerarin group(P

Objective To investigate potential protection by protect against ethanol-induced apoptosis of spermato-genie cells in rat testis. Methods Thirty SD adult male rats were randomly divided into three groups: normal group, alcohol group and puerarin group. At 40th day, BCL-2 and BAX of spermatogenic cells of testis tissue were checked by RT-PCR and immunohistochemistry; Apoptosis of spermatogenic cells was determined by TUNEL. Re-suits The results of RT-PCR and immunohistochemistry indicated that BCL-2 and BAX of spermatogenie cells were not significanty different between puerarin group and normal group, but there was the significant difference between alcohol group and puerarin group (P <0.01). Apoptosis of spermatogenic cells in alcohol group was significantly higher than normal group. Conclusion Spermatogenic cells could generate apoptosis by changing the expression of BCL-2 and BAX. Puerarin could inhibit this damage of didymus by alcohol.

AIM:To observe the effect of spironolactone on myocardial fibrosis of spontaneously hypertensive rats(SHR).METHODS:Sixteen fourteen-week-old male SHRs were randomly assigned to spironolactone and SHR group equivalently(n=8).Rats in each group were given 30 mg ? kg-1 ? d-1 spironolactone and equal sodium chloride respectively for 12 weeks by gavage.Eight fourteen-week-old male SD rats were as control group.Connective tissue growth factor(CTGF),transforming growth factors beta-1(TGF?1),collagenⅠand Ⅲ were measured by qualitative and semiquantitative immunohistochemical staining and semiquantitative reverse transcription polymerase chain reaction.Masson staining was used to determine total collagen in left ventriculum.Alkaline hydrolysis method was used to detect the concentration of hydroxyproline(Hypro)in the myocardium of left ventricle.RESULTS:Left ventricular index(LVI),collagen volume fraction(CVF),Hypro and the expression of TGF?1,CTGF,collagenⅠand Ⅲ in SHR group were significantly higher than those in SD group(P

AIM: To investigate the effects of intracellular free calcium ([Ca 2+ ]i) from different resources on the proliferation mediated by mitogen activated protein kinase (MAPK) in vascular smooth muscle cells (VSMCs). METHODS: Cultured VSMCs were used in all experiments. Calcium influx was stimulated by angiotension Ⅱ(AngⅡ). The release of intracellular calcium stores was induced by inositol trisphosphate(IP 3) and ryanodine(RY). MAPK activity was measured by [?- 32 P]-ATP incorporation , MAPK protein expression by western blot, VSMCs proliferation by -Leucine(-Leu) and -Thymidine(-TdR) incorporation. RESULTS: Compared to the control VSMCs, AngⅡ, IP 3 and RY significantly increased [Ca 2+ ]i concentration , activity of MAPK and its protein content in VSMCs. The promotion of -Leu and -TdR incorporation in VSMCs was also observed ( P

Objective To investigate the impact on renal fibrosis by inhibition of connective tissue growth factor( CTGF) by RNA interference in spontaneous hypertension rat( SHR) . Method Twenty SHR were randomly (random number) divided into SHR group ( n = 10) and RNAi group ( n = 10), eight Wistar-Kyoto rats were set as control. At the end of RNA interference procedure, all the rats were sacrificed and the kidneys were harvested. The mRNA and plasmosin of CTGF and fibronectin(FN) of renal tissue were extracted and measured by RT-PCR and Western Blotting. And the localization of CTGF and FN were analyzed with immunohistochernistry technique. The collagen deposition(shown as collagen volume traction, CVF) were evaluated with 0.1% sirius-picric staining, and the hydroxyproline of myocardium were detected by colorimetry. Results The mRNA and protein expression of CTGF decreased 66% and 62% in RNAi group (P < 0.01). The mRNA and protein expression of FN decreased 56% and 51% in RNAi group.The same inhibition effect was observed by hislological analysis. Immuno-histochemistry showed that CTGF localized both in renal parenchyma and renal interstitium, whereas FN majorly expressed in renal interstitium. Observation with light microscope showed that collagen deposition(CVF)decreased sharply in RNAi group versus SHR group. And the same effect was viewed in hydroxypnoline assay[SHR group: (0.596 ± 0.067) μg/mg, RNAi group: (0.368±0.084) μg/mg, P < 0.01 ] .Further study by polarized microscope displayed that RNA interference mainly suppressed type I collagen synthesis. Conclusions Targeted inhibition of CTGF by RNA interference leads significant decrease of extracellular matrix deposition in kidney. And the anti-fibrotic effect independent of lower the blood pressure. This study indicated CTGF take a key role in the development and progress of renal fibrosis.

Objective To find the cells in myocardium controlling in cardiomyogenic differentiation of marrow derived-mesenchymal stem cells(MSCs).Methods Rat marrow derived-MSCs,cardiomyocytes(CM)and endothelial cells(EC)were isolated and cultured,then MSCs were incubated with the CM or EC or cultured alone after labeled by 5-Bromo-2-deoxyuridine(BrdU)for 72 hours.The cells were identified by morphology,immunocytochemistry and double immunocytochemistry.Results More than 95% of the cells isolated and cultured belonged to MSCs,CM or EC.Incubated with CM,most MSCs labeled by BrdU showed cardiomyocyte-like morphology.After 4 weeks of cultivation,positive staining of double immunocytochemistry of BrdU and Sarcomeric Actin or Connexin-43(CX-43)were observed in 44% cells.However,there was no obvious change in morphology and no positive staining of double immunocytochemistry was recorded in MSCs incubated with EC or cultured alone after 1、2、3 and 4 weeks of cultivation.Conclusion MSCs can differentiate into cardiomyogenic cells in cardiomyocyte microenvironment and cardiomyocytes may play a crucial role in the differentiation.

Objective To explore the effects of Shengmai Injection on tumor growth and the expression of P-gp in transplanted tumor of human gastric carcinoma of SGC7901/VCR cell in nude mice.Methods Transplanted tumor model of human gastric carcinoma of SGC7901/VCR cell in nude mice was built, which was divided randomly into five groups: normal saline control-group, 5-FU group, 5-FU + verapamil group, 5-FU +Shengmai group, Shengmai group. Nude mice growth state was observed, average weigh and inhibition rate of transplanted tumor were calculated, and the expression of P-gp was detected.Results There was significanf difference in terms of transplanted tumor weight,volume among 5-FU+Shengmai group and 5-FU group and normal saline group(P0.05); P-gp express had difference between normal saline group and shengmai group, P