Objective In order to explore the biological features of bone marrow mesenchymal stem cells. Methods The ALPase activity, Vimentin and Laminin were detected by immunocytochemistry. Cell cycle and surface antigenic features were analyzed by flow cytometry technique. Cell growth curve was measured by MTT test. The telomerase activity was detected by in situ hybridization. Results The ALPase activity and Laminin were negative, Vimentin was positive in the MSCs. The percentage of G1, S, and G2 of cell cycle was 89.5%,7.8%,7.7%, respectively. MSCs were positive for CD166、CD29、CD44、CD105, and negative for CD34 and HLA-DR. The average doubling time of cells was 32 hours. The telomerase activity was positive. Conclusion The cultured cells were not hemopoietic stem cells or fibroblasts, but the MSCs that had not differentiated. Furthermore, it had relatively high proliferation ability.

Objective To investigate the feasibility of the committed differentiation of mesenchymal stem cells(MSCs) into tenocytes,the present study was carried out. Method NMP12 gene transfer to MSCs was performed with pEGFP C1 expressing vector by electroporation. Results Green fluorescence protein(GFP) expression was detected as early as 6h postelectroporation, peaked at 12h and could maintain 3 days. And then, GFP expression of some cells was weakened. Conclusion The results of the present study indicated that the successful transfection of MSCs was made, an the committed differentiation of MSCs into tenocytes was practicable

BACKGROUND: Growth differentiation factor-5 (GDF-5) plays an important role in the development and formation of cartilage, extremities, and joints, which is a widely used joint development marker.OBJECTIVE: To express mature peptide of human GDF-5 in E. coil by the way of genetic engineering, and to explore the inductive activity of recombinant protein in vivo.DESIGN, TIME AND SETTING: The observation experiment based on gene was performed at the Analysis and Testing Center of Southern Medical University from January to June 2006.MATERIALS: Human fetus cartilage tissue was harvested from Department of Gynaecology and Obstetrics, and the consent was obtained from the family. Ten KM mice were purchased from experimental animal center of Southern Medical University, half male and half female, weighing 18-22 g, aged 6-8 weeks.METHODS: The hGDF-5 gene encoding mature peptide was gained by RT-PCR from the total RNA which was extracted from fetus cartilage tissues, and was inserted into the pET22b(+) vector to construct recombinant prokaryotic expression plasmid pET22b(+)-GDF5, which was transformed into E. coil BL-21 to be expressed after IPTG induction. Proteins of interest were purified with sepharose chelated with nickel ions (Ni2+) and then implanted in mouse hindlimb muscle to evaluate the biological activities by routine hematoxylin-eosin staining.MAIN OUTCOME MEASURES: The expression, sequencing of target gene was observed by agarose gel electrophoresis, and the protein expression was detected by SDS-PAGE electrophoresis, meanwhile, the GDF5-inducing activity was evaluate by histological observation.RESULTS: RT-PCR product was about 350 bp in length, which was confirmed by double enzyme digestion of the recombinant plasmid, sequencing result was in agreement with the reported hGDF-5 sequence in Genbank. SDS-PAGE analysis showed a conspicuous band representing a new foreign protein with relative molecular mass of approximately 14 KD after induced expressioin. Cartilage tissues were formed in the mouse muscle where the purified proteins were implanted. CONCLUSION: The integral human GDF-5 mature peptide gene was cloned successfully from human fetus cartilage tissue and a high-yield expression was achieved in E. coli, the pudfied protein has chondrogenic activities in vivo.

Objective We established the animal models of obesity induced by high-fat diet, in order to study the mRNA and protein expression of regulation molecules related with iron metabolism about hepcidin, lipocalin-2 ( LCN2 ) , ferroportin-1 (FPN1) in obese mice’ s liver and the molecular regulation mechanism.Methods C57BL/6J (4 ~6 weeks) mice were randomly divided into control group and obesity model group, each group of ten.The obesity group were fed with a high-fat diet and the control group were given the normal diet for lasting 15 weeks.After we successfully established the obesity animal model, the expression level of hepcidin, LCN2 and FPN1 mRNA in the liver were measured by Real-time fluorescent quantitative PCR method and the protein expression level of LCN2 and FPN1 were measured by Western-Blot.Results Compared with the control group, the expression level of hepcidin mRNA in the liver was increased in obesity group (P <0.05), however, the expression level of LCN2, FPN1 was no significant difference (P >0.05).Conclusion Obesity can increase the expression of hepcidin mRNA, however, there was no significantly effect on the expression of LCN2, FPN1.So, we can’t think that obesity can affect the expression of LCN2 and FPN1, lead to the ability of cells uptake and release iron abnormal, then appear iron metabolism disorders.As a result, leading to iron deficiency.Maybe obesity can affect other regulatory molecules related with iron metabolism through up-regulation the expression of Hepcidin or the more complex regulatory mechanisms.We still need further experimental research and exploration.This research also provides the basis of theoretical and experimental for the further study the effects of obesity on the expression of regulation molecules related with iron metabolism in obesity mice’ s liver and the mechanism of iron deficiency.

Objective To observe the effect of acupoint magnetic therapy on coronary heart disease.Methods 80 patients with coronary heart disease were divided into treatment group( n =40) and control group( n =40).The treatment group was treated with acupoint magnetic therapy on the basis of conventional treatment,and the control group with conventional treatment.Clinical therapeutic effect,the scores of traditional Chinese medicine symptom were observed.Results There was significant difference between the two groups in the total effective rate and the scores of traditional Chinese medicine symptom[90.0％ vs 72.5％,(13.25 ± 3.68 ) vs (15.18 ± 4.16),all P ＜0.05].Conclusion Acupoint magnetic therapy for treatment of coronary heart disease has an obvious therapeutic effect.

【Objective】To observe the anti-stress effect of koumine(Kou)on mice.【Methods】Kunming mice were randomized into model group,low-,moderate-and high-dose koumine(1.2,2.4,and 4.8 mg?kg-1?d-1)groups.The treatment lasted 7 days.One hour after last administration,stress tests such as weight-bearing swimming,antihypoxia,high-temperature resistance and low-temperature resistance were carried out.The changes of serum superoxide dismutase(SOD)activity and malondialdehyde(MDA)content were also observed.【Results】During the anti-stress tests,the survival time was prolonged in koumine groups as compared with the model group(P0.05).【Conclusion】Koumine can increase the mice tolerance of weight-bearing swimming,cold and hypoxia,and its anti-stress mechanism may be related to the antilipid peroxidation.

Comparative medicine is a new crossing frontier branch subject based on laboratory animal science,and the content,aim and request of its research have been remarkably different from that of laboratory animal science.Acting as the methods,means and support conditions of advancment and development of medicine research,comparative medicine is an important platform ensuring the sustainable development of medicine,and is imperative for the medico to be educated with systemic comparative medicine.

Objective:To study the anti proliferation and inducing apoptosis effects of cytokine induced killer cells CIK cells on MGC 803 gastric cancer cell lines and to probe its underlying mechanism.Methods:To detect the anti proliferation and the cytotoxicity of CIK cells on MGC 803 gastric cancer line by MTT assay.The morphological changes of the apoptosis cell were observed by HE stain, scanning and transmission electron microscope. The positive expression of p53, p16,C myc were determined by immunocytochemistry (ICC).Results:MTT assay showed that the inhibitive rate inhanced obviously with the addition of Effect/Target rate and extension of time ( P

Objective To observe the influence of oleanolic acid on the ethology of 9-month-old mice,the completeness of synapsis structure and the expression of cAMP-response element binding protein (CREB) in cortex and hippocampus.Methods Thirty 9-month-old healthy male SMAP8 mice were randomly divided into model group,oleanolic acid group and aricept group,and with 10 rats in each group,while 10 healthy male mice of the same age and species as normal group.Oleanolic acid group and aricept group were given intragastric administration with corresponding drugs,while the normal group and model group were given the same amount of physiological saline.4 weeks later,the ethology changes were observed by Morris water maze and morphology changes of hippocampus neurons were viewed by electron microscope and the expression of CREB was detected by Western Blot.Results (1)Morris water maze results suggested that compared with the normal group,the latency time in the model group mice was longer,which were ((83.33±4.96) s,(75.13±6.01) s,(71.75±7.77) s,(63.40± 8.93) s,(60.97±8.38) s),while compared with the model group,the latency time in the oleanolic acid group and the aricept group was remarkably shorter (P＜ 0.05),which were (75.97± 4.49) s,(64.98± 4.93) s,(64.16± 6.23) s,(53.47±5.99) s,(47.91±7.64) s and (71.30±7.65) s,(63.32±7.57) s,(59.82±4.69) s,(52.28±5.90) s,(46.22±7.27) s respectively.In the spatial probe trial,compared with tbe normal group,the crossing times of the model was less,while compared with the model one,the crossing times of the oleanolic acid group and the aricept group was more(P＜0.05).(2)Compared with the normal group,the number of synapses in the model group was smaller,in which the synaptic cleft was mixed with the front of synapses severely swollen,uninform synaptic vesicles and few clear outlines of mitochondrias.While the oleanolic acid group and the aricept group had clear synapses outlines with the front of synapses slightly swollen,intensive and uniform synaptic vesicles and clear mitochondrias with their cristae not easy to be seen.(3) The Western Blot showed that compared with the normal group,there was a decline in the CREB expression both in the cortex and hippocampus in the model group,while compared with the model group,there was a rise in the oleanolic acid group as well as the aricept group(P＜0.05).Conclusion Oleanolic acid can improve the learning and memorizing of model rats,which is possibly related to the increased expression of CREB protein to protect the synapses structure of model mice.

BACKGROUND:Pathophysiological mechanisms after spinal cord injury are very complex, so there is no compressive and in-depth understanding on it.
OBJECTIVE:To study the effect of dura mater spinalis integrity on cytokine levels in the cerebrospinal fluid of animal models of spinal cord injury.
METHODS:The white rabbit models of spinal cord injury were established using clamp compression method, and then the models were randomly divided into four groups:no dura mater spinalis defect group, dura mater spinalis defect group, dura mater spinalis defect composite with membrane repairing group and dura mater spinalis defect composite with autologous fascia repair group. Enzyme-linked immunosorbent assay was
performed to detect the changes of levels of cytokines (interleukin-6, interleukin-10 and tumor necrosis factorα) in the cerebrospinal fluid at 30 minutes, 1, 3, 6, 12 and 36 hours after surgery.
RESULTS AND CONCLUSION:The levels of interleukin-6, interleukin-10 and tumor necrosis factorαin the
cerebrospinal fluid of the dura mater spinalis defect group, dura mater spinalis defect composite with membrane repairing group and dura mater spinalis defect composite with autologous fascia repair group were significantly lower than those of the no dura mater spinalis defect group at 6 hours after surgery (P<0.05). There were no significant differences in the levels of interleukin-6, interleukin-10 and tumor necrosis factorαat other time points between groups (P>0.05). The results indicate that maintaining the integrity of dura mater spinalis of the spinal cord injury model can affect the levels of interleukin-6, interleukin-10 and tumor necrosis factorαin the
cerebrospinal fluid, thus inhibiting the inflammatory response.