AIM　To explore the effect of berberine in inhibition of proliferation and induction of apoptosis of gastric cancer cell MGC-803.METHODS　The MGC-803 cell morphology and cell counting were studied by Trypan blue, MTT, Methyl green pyronin stain assay, the MGC-803 cell DNA changes were investigated by flow cytometry and agarose gel electrophoresis.RESULTS　Berberine could significantly inhibit the growth of MGC-803. After 48 hours of treatment with different concentration berberine (8, 4, 2, 1 mg*L-1), the inhibitive rate of MGC-803 were 86.7%, 65.9%, 20.0%, 0%, respectively. Meanwhile, MGC-803 showed typical apoptosis features: cell shrinkage, nuclear condensation, DNA fragmentation and formation of apoptotic bodies. A typical subdiploid peak before G0/G1 phase was observed by flow cytometry. The Trypan blue stain achromatophilia rates of death cells, which with well membranes, were between 60%～82%. Agarose gel electrophoresis analysis, which showed that the chromosomes of MGC-803 were broken by berberine before the structures of membrane were damaged, while DNA broke into long fragments rather than small fragments, supported that the activation effect of berberine on nuclease was mild.CONCLUSION　Berberine could significantly inhibit proliferation of gastric cancer cell MGC-803 and induced apoptosis, and the cytotoxicity of berberine on MGC-803 was weak.

Many tumor cells are resistant to cell apoptosis through the expression of antiapoptotic proteins. c-FLIP is a ma-jor resistance protein of antiapoptosis. In human cells, there are three types of c-FLIP, c-FLIPL , c-FLIPS and c-FLIPR . The c-FLIP binds to FADD to prevent the formation of procaspase-8-DISC and the subsequent activation of caspase cascade. Further-more, c-FLIPL and c-FLIPS have multifunctional roles in various cellular signaling pathways, as well as up-regulating several cyto-protective signaling. Studies show that upregulation of c-FLIP has been found in various tumors, and its downregulation has been shown to restore apoptosis triggered by various chemothera-peutic agents, like the transcriptional regulating agents, trichos-tatin-A, camptothecin, cisplatin, doxorubicin, etc. or other new biotechnologies, such as the specific siRNA. Therefore, c-FLIPS are important targets of cancer therapy. This review summarizes the results on the role of c-FLIP in cancer chemotherapy of tradi-tional antitumor agents and siRNA, and to provide new ideas and rationales of searching for the antiapoptosis effective compounds that can specifically antagonize c-FLIP.

AIM To explore the effect of berberine in inhibition of proliferation and induction of apoptosis of gastric cancer cell MGC 803.METHODS The MGC 803 cell morphology and cell counting were studied by Trypan blue, MTT, Methyl green pyronin stain assay, the MGC 803 cell DNA changes were investigated by flow cytometry and agarose gel electrophoresis.RESULTS Berberine could significantly inhibit the growth of MGC 803. After 48 hours of treatment with different concentration berberine (8, 4, 2, 1 mg?L -1 ), the inhibitive rate of MGC 803 were 86 7%, 65 9%, 20 0%, 0%, respectively. Meanwhile, MGC 803 showed typical apoptosis features: cell shrinkage, nuclear condensation, DNA fragmentation and formation of apoptotic bodies. A typical subdiploid peak before G 0/G 1 phase was observed by flow cytometry. The Trypan blue stain achromatophilia rates of death cells, which with well membranes, were between 60%～82%. Agarose gel electrophoresis analysis, which showed that the chromosomes of MGC 803 were broken by berberine before the structures of membrane were damaged, while DNA broke into long fragments rather than small fragments, supported that the activation effect of berberine on nuclease was mild.CONCLUSION Berberine could significantly inhibit proliferation of gastric cancer cell MGC 803 and induced apoptosis, and the cytotoxicity of berberine on MGC 803 was weak.

Objective To explore the effect of acupoint injection of bone marrow mesenchymal stem cells (BMSCs)on hemodynamics of rat model with acute myocardial infarction(AMI). Methods Sixty Sprague-Dawley (SD)rats were randomly divided into sham group,model group,myocardium injection BMSCs group and acupoint injection BMSCs group(each n=15). The left anterior descending coronary artery(LAD)was ligated to establish a rat model of AMI. After the rat model was successfully established for 72 hours,0.2 ml BMSCs(1×1010/L)were transplanted by a micro-quantity syringe at 6 points in equal amount in the LAD blood-supply area and its periphery in the myocardial injection group,while in the acupoint injection group,0.3 ml BMSCs(1×1010/L)was transplanted at each of the following acupoints:Xinshu,Zhiyang and Tanzhong. Four weeks after AMI,polyethylene tubing was inserted into the right carotid artery to measure the hemodynamics,at the same time animals were sacrificed,and the heart was take out to calculate the heart mass index(HMI)and left ventricular mass index(LVMI). Results One, 3,5 and 4 rats were respectively dead in the sham group,model group,myocardium injection group,and acupoint injection group during the experimental period. Compared with the sham group,the left ventricular systolic pressure (LVSP,mm Hg,1 mm Hg=0.133 kPa),the maximum rate of increase of left ventricular pressure(+dp/dt max, mm Hg/s), the maximum rate of decrease of left ventricular pressure(-dp/dt max,mm Hg/s), the maximum logarithmic change rate of left ventricular pressure〔(dp/dt)?P-1max,s-1〕,HMI(mg/g),LVMI were significantly decreased,and left ventricular end-diastolic pressure(LVEDP,mm Hg),heart rate(HR,bpm)were obviously increased in model group(all P<0.05). Compared with the model group,the LVSP,+dp/dt max,-dp/dt max, (dp/dt)?P-1max, HMI,LVMI were significantly increased in myocardial injection group and acupoint injection group〔LVSP:130.38±14.96,124.36±14.36 vs. 114.36±12.71,+dp/dt max:4707.52±394.36,4597.14±411.05 vs. 3791.43±327.29,-dp/dt max:4075.11±317.89,3938.05±373.76 vs. 3116.32±275.04,(dp/dt)?P-1max:215.26±21.29,197.39±18.96 vs. 155.93±25.14〕,and the LVEDP and HR were significantly decreased(LVEDP:5.15±2.39,5.64±1.96 vs. 10.58±2.49,HR:400.50±42.58,395.55±44.62 vs. 414.51±35.75,all P<0.05). There were no significant differences in above indexes between myocardium injection group and acupoint injection group. Conclusion Acupoint injection of BMSCs can improve the heart function of rat model with AMI.

[ ABSTRACT] AIM:To investigate the effects of maternal limb ischemic preconditioning ( LIP) on the mitochon-drial structures and functions of the hippocampal neurons induced by reoxygenation in the intrauterine distress fetal rats. METHODS:Pregnant rats (n=40) were randomly divided into 4 groups: sham (S) group, LIP group, fetal distress ( FD) group and LIP+FD group.Intrauterine ischemia model was established through the experimental design.The ultra-structure of the mitochondria in CA1 area of the hippocampus was observed .The mitochondrial membrane potential and re-active oxygen species ( ROS) were measured .The content of ATP and MDA in the hippocampus tissue was detected.The activity of Mn-SOD was observed.RESULTS:Compared with sham group, the ultrastructure of mitochondria in CA1 area of the hippocampus was damaged in FD group and LIP+FD group.The mitochondrial membrane potential, the content of ATP and the activity of Mn-SOD were decreased.However, the content of ROS and MDA was increased.Compared with FD group, the ultrastructure of mitochondria in CA1 area of the hippocampus was intact in LIP+FD group.Furthermore, the reduced mitochondrial membrane potential and ATP content were inhibited.The activity of Mn-SOD was increased, but the content of ROS and MDA was decreased in LIP+FD group.CONCLUSION:Limb ischemia preconditioning inhibits the damage the mitochondria of fetal hippocampal neurons induced by reoxygenation in the intrauterine distress fetal rats.

Objective To compare the pharmacological actions of Liandai Tablet(LT) and its main active components on apoptosis of gastric cancer cells and DNA damage. Methods Effects of Liandai Tablet(LT) and its main active components,berberine and indirubin,on growth and apoptosis of gastric cancer cell strain MGC_803 were explored and their effects on DNA damage were also studied. Results LT serum in high and low dosages and berberine could inhibit the growth of MGC_803 as compared with the control group,and typical morphological features of apoptosis were found in the MGC_803 by methyl green pyronin stain assay.But indirubin at various concentrations showed no obvious inhibitory effects. Agarose gel electrophoresis assay revealed that the MGC_803 cell DNA was split into large fragments when treated with berberine. Conclusion LT serum exerts a similar inhibitory effect on the growth and apoptosis of gastric cancer cells as compared with berberine.The effects of LT at various serum concentrations on MGC_803 DNA was less than that of berberine,and indirubin at the given concentration had no this effect.

BACKGROUND: Stem cell transplantation can significantly improve heart function foUowing myocardial infarction. This is correlated with the differentiation of stem cells into cardiomyocytes and promotion effect on angiogenesis. Paracrine and ventricular reconstruction inhibition (especially extracallular collagen reconstruction) have important effects on improving heart function.OBJECTIVE: To investigate the effects of bone marrow mesenchymal stem cell (BMSC) transplantation on coUagen remodeling after acute myocardial infarction in rabbits.DESIGN, TIME AND SETTING: The randomized controlled animal study was performed at the Laboratory of Acupuncture and Electrophysiology of Liaoning University of Traditional Chinese Medicine from June to August 2007.MATERIALS: A total of 57 healthy Japanese rabbits were purchased from Experimental Animal Center, Uaoning University of Traditional Chinese Medicine.METHODS: BMSCs were acquired from the bone marrow of two rabbits, and marked with BrdU before transplantation. Ten rabbits served as a normal group. Forty-five rabbits were used to establish the left ventricular infarct by ligation of the left coronary artery. Thirty success models of myocardial infarction were randomly divided into 3 groups (n=10)" model, saline and call transplantation groups. Following 7 days of myocardial infarction, rabbit models in the cell transplantation group were injected in the ear vein with 1 mL of BMSCs (2x106 cells). Rabbits in the saline group were infused with 1 mL of saline. The culture was performed for 5 weeks.MAIN OUTCOME MEASURES: Fibrous structure of myocardial stroma was observed, and collagen volume fraction was measured by Masson Trichrome staining. The ratio of type Ⅰ and Ⅲ collagen was determined by immunohistochemistry.RESULTS: BrdU-positive BMSCs could be seen in the cell transplantation group. After myocardial infarction, a few collagen fibers was confluent in or surrounding the infarct area, arranged orderly in the cell transplantation group. Collagen fiber plaque-shaped confluence was significant, and arranged disorderly in the model and saline groups. At 5 weeks following myocardial infarction, compared with the normal group, collagen volume fraction was significantly decreased in and surrounding the infarct region (P < 0.05), and the ratio of type Ⅰ and Ⅲ collagen was increased significantly in the model group (P < 0.05).Compared with the model group, collagen volume fraction and the ratio of type Ⅰ and Ⅲ collagen were significantly decreased (P< 0.05).CONCLUSION: BMSCs could survive in infarct heart. BMSCs transplantation could reduce collage volume and improve collage ratio and had beneficial effects on collage remodeling processes after acute myocardial infarction.

Objective To investigate the effect of maternal limb ischemic preconditioning on the expression of caspase-3 in neurons in brain tissues after reoxygenation in the fetal rats with intrauterine distress.Methods Forty Sprague-Dawley rats at 19 days of gestation were randomly divided into 4 groups (n=10 each) using a random number table:sham operation group (group S),limb ischemic preconditioning group (group LIP),fetal rat distress group (group FD),and limb ischemic preconditioning + fetal rat distress group (group LIP+FD).Distress/reoxygenation model was established by clamping the uterine and ovarian arteries and veins with a micro-artery clamp for 15 min followed by removal of the clamp to permit reperfusion.Limb ischemic preconditioning was induced by 3 cycles of occlusion of the lower limb blood flow at the site of the right groin for 5 min with a tourniquet followed by 5 min unclamping.In group LIP+ FD,the uterine and ovarian arteries and veins were clamped,and limb ischemic preconditioning was performed at the same time.Cesarean section was performed on 2 days after the end of treatments in each group,and the fetal rat mortality rate was calculated.The fetal rats alive were sacrificed,and the hippocampi were isolated for determination of neuronal apoptosis (by TUNEL) and expression of caspase-3 protein and mRNA (by Western blot and real-time reverse transcriptase polymerase chain reaction,respectively) in hippocampal CA1 region.Apoptosis index was calculated.Results Compared with group S,the fetal rat mortality rate and apoptosis index were significantly increased,and the expression of caspase-3 protein and mRNA in hippocampal CA1 region was significantly up-regulated in FD and LIP+FD groups (P＜0.05 or 0.0l),and no significant change was found in the parameters mentioned above in group LIP (P＞0.05).Compared with group FD,the fetal rat mortality rate and apoptosis index were significantly decreascd,and the expression of caspase-3 protein and mRNA iu hippocampal CA1 region was significantly down-regulated in group LIP+FD (P＜0.05 or 0.01).Conclusion The mechanism by which maternal limb ischemic preconditioning inhibits apoptosis in neurons after reoxygenation is related to down-regulation of the expression of caspase-3 in the fetal rats with intrauterine distress.

BACKGROUND:Now, the bone marrow mesenchymal stem cel homing is thought to be mediated by adhesion molecules and chemokines, and this process involves bone marrow endothelial cel s, hematopoietic stem cel s, bone marrow microenvironment and its secreted or expressed molecules, in which, adhesion molecules may play an important role.
OBJECTIVE:To explore the migrating and chemotactic mechanism of bone marrow mesenchymal stem cel s via acupoint injection into myocardial cel s by determining the expression of vascular cel adhesion molecule-1 and very late antigen-4.
METHODS:Bone marrow mesenchymal stem cel s were cultured using adherent method, and the passage 3 cel s were used as seed cel s at a density of 1×1010/L. Sixty Sprague-Dawley rats were randomly divided into sham group, model group, myocardial injection group, and acupoint injection group, 15 rats in each group. The left coronary arteries of rats were ligated for establishing a model of myocardial infarction. At 72 hours after myocardial infarction, 0.3 mL bone mesenchymal stem cel s were transplanted into the Xinyu, Zhiyang, Tanzhong acupoints, respectively, in the acupoint injection group;while in the myocardial injection group, secondary thoracotomy was done, and 1.2 mL bone marrow mesenchymal stem cel s were equably transplanted into six
sites in the feeding area of the left anterior descending artery and the surrounding myocardium. At 4 weeks after myocardial infarction, a multi-channel polygraph was adopted for detection of hemodynamic parameters, and the levels of serum vascular cel adhesion molecule-1 and very late antigen-4 were measured by ELISA.
RESULTS AND CONCLUSION:The heart function of rats in the myocardial injection and acupoint injection groups were improved, and compared with the model group, the levels of serum vascular cel adhesion molecule-1 and very late antigen-4 were significantly higher in the myocardial injection and acupoint injection groups. However, there was no significant difference between the myocardial injection and acupoint injection groups. These findings suggest that vascular cel adhesion molecule-1 and very late antigen-4 may be one of the chemotactic mechanisms of bone mesenchymal stem cel s transplanted in myocardial infarction model rats by acupoint injection.

The cloning and identification of frc gene from Oxalobacter formigenes in the intestines of Chinese people were conducted. The genomic DNA of Oxalobacter formigenes was extracted. frc gene fragment was amplified by polymerase chain reaction (PCR) and linked with pEGFP-C1. The recombinant plasmid was designated pEGFP-frc and was identified by restriction-enzyme digestion and sequencing. Human embryo kidney 293 cells were transfected with pEGFP-frc, then RT-PCR and Western blotting were performed to detect the expression of frc gene. The length of frc gene was found to be 1287 bp, and the homology of nucleotides and amino-acid residue with the sequence in GenBank was 95.88% and 99.07%. Bright green fluorescent light could be observed in 293 cells transfected with the pEGFP-frc. frc mRNA and fusion protein FCoAT-EGFP were detected in the cells. It is concluded that frc gene cloned from the Oxalobacter formigenes in the intestines of Chinese people can be expressed in eucaryotic 293 cells and keep its enzyme activity.