The experiment was conducted on the actions of 5 kinds of reagents on HL - 60 cellular wither, viz. Yisuiling chloroform extract, rabbit serum chloroform extract, rabbit liver paste chloroform extract, Etoposide, and normal rabbit serum. Results revealed that after fluid incubation for a short period, the HL-60 cultured cells showed no wither in the former 4 groups under microscope and flowing cytometer. Among them, action was strongest in rabbit liver paste of Yisuiling extract group, similar to the positive control of Etoposide. Followed by rabbit serum, and the action was weak in Yisuiling chloroform extract. There was no obvious action of induced wither by normal rabbit serum. The mechanism of induced wither is still under investigation.

AIM:To investigate effect of recombinant human thrombopoietin on exsanguine thrombocytopenia mice. METHODS:Normal peripheral platelet counts were performed on sample obtained from the tail vein of purebred Babl/c mice including experimental and control groups before experimentation. rhTPO was injected into the mice by intraperitoneal injection once a day for 7 days. On the seventh and the fourteenth day, the mice were phlebotomized from the supra-obitalis vein in order to make exsanguine thrombocytopenia animal model. At the same time, we observed the biological activity of recombinant human thrombopoietin in vivo and the mice's death rate. RESULTS: On the seventh day and the fourteenth day, platelet counts of mice treated by rhTPO were higher than those by PBS (P＜0.05). Moreover the platelet counts of mice in experimental group of rhTPO showed increasing tendency following experimental days. In addition, death happened in two groups after those mice were phlebotomized from the supra-obitalis vein, but the death rate in negative control group was evidently higher than that in experimental group (P＜0.05). CONCLUSION:rhTPO had obvious biological activity in increasing platelet production, which resulted in the drop in thrombocytopenia mice's death rate.

BACKGROUND:There are myoblasts in human embryonic skeletal muscle. It remains poorly understand whether myoblasts in vitro can form myotube and what are the corresponding markers for identifying myoblasts and myotubes. OBJECTIVE:To investigate whether in vitro cultured myoblasts from human embryonic skeletal muscle can form myotube and whether they can express neural markers. METHODS:Human embryonic muscle-derived myoblasts were cultured in serum-containing medium. When the primary culture was established, cultured cel s were identified with immunocytochemistry for neural markers, such asβ-tubulin markers (desmin, myogenin, smooth muscle actin and myosin). RESULTS AND CONCLUSION:A population of myoblasts could migrate from human embryonic muscle tissues. They could express the markers for skeletal muscle such as desmin and myogenin, and they could express neuron specific enolase, nestin and neurofilament 200. They could form myotubes in vitro, and myotubes expressedβⅢ-tubulin, neurofilament 200 and glial fibril ary acidic protein. The data support the hypothesis that myoblasts from human embryonic muscle express neural markers and muscle markers, and cultured myoblasts and myotubes expressed neuron specific enolase,β-tubulin Ⅲ, nestin, neurofilament 200 and glial fibrillary acidic protein. This indicates that these markers could not be used for cel identification of trans-differentiation study from muscle origin to nervous system.

Objective: To investigate the optimal culture conditio ns for adipose tissue-derived stromal cells(ADSCs) and for the induction of these cells to differentiate into osteogenic cells. Methods: ADSCs were cultured with routine methods,bFGF at 20 ng/ml was added into the medium and the proliferative of ADSCs was examined by cell counting. 0.1 ?mol /L of dexamethasone,10 mmol/L of ?-glycerophosphate and 50 ?mol/L of ascorbic acid were adapted to induce the cells to differentiate into osteogenic cells, ADSCs were identified by immunocytochemistry and differentiated osteogenic cells were identified by alkaline phosphatase(AP) staining and immunocytochemistry. Result: A population of ADSCs could be isolated from adul t human adipose tissue,the cells were fibroblast-like and could be maintaine d in vitro for extended periods with stable population doubling.The cells w ere expanded as undifferentiated in culture for more than 10 passages, indicati ng their proliferative capacity.bFGF stimulated the cell proliferation.Dexameth asone,?-glycerophosphate and ascorbic acid induced (40?8.6)% of ADSCs to ex press alkaline phosphatase(AP) ,(35?10.6)% of AP positive ADSCs were found to be collagen I positive. Calcification plaques were occasionally found in the cul tures. Conclusion:The data support the hypothesis that adu lt human adipose tissue contains stem cells capable of diffferentiating into ost eogenic cells.

AIM:To investigate the potential of differentiatng into myocytes of the granulocyte colony-stimulating factor(G-CSF)-mobilized CD34 + cells. METHODS: Three hours after intraperitoneal injecction of isoprenaline(ISO) to develop acute ischemic model, rats' bone marrow hematopoietic stem cells were mobilized to the site of myocardial infarction by G-CSF. The techniques of immunohistochemisty and HE stain were used to detect the infiltration of CD34 + cells and the regeneration of myocytes in the infarct zones. RESULTS: 24 hours after administration of ISO , a large amount of infiltrative monocytes and regenerative myocytes which were CD34 positive expression could be found in the infarct zones of the G-CSF treatment group, while majority of the infiltrative inflammatory cells in control group were neutrophils and there was no infiltrative cells and myocytes which were CD34 positive expressio, 2 weeks after administration of ISO, there were a plenty of scar in control group, but not in the G-CSF treatment group. CONCLUSION: G-CSF-mobilized CD34 + cells possess the potential to differentiate into myocytes and it may be used in treating acute myocardial infarction.

ObjectiveTo investigate the value of combined four-chamber view,left and right ventricular outflow tract view and three-vessel view of two-dimensional echocardiography (2DE) in prenatal screening for fetal congenital heart disease (CHD). MethodsFour combined views of 2DE were used to detect fetal hearts in 2419 fetuses at 21～ 25 gestational weeks.The echocardiograms were performed on all 2382 live-birth infants.Chi-square test was applied for statistical analysis.Sensitivity,specificity,positive predict value and negative predict value were calculated. Results The prevalence of fetal CHD was 11.62％ (281/2419).Among the 281 CHD fetuses,87.18％ were simple CHD (n=245) and 12.82％ were complex CHD (n=36).No difference was found in the positive rate of fetal CHD between the high-risk group and non-high-risk group [13.60％(34/250) vs 11.39％(247/2169),x2=1.069,P＜0.05].Thirty-six cases of CHD could be detected by the four combined views in prenatal screening with the sensitivity,specificity,positive and negative predictive value of 12.8％,99.8％,90.0％ and 89.7％,respectively.However,the diagnostic sensitivity of four combined views for simple CHD was 2.9％(7/245) and 80.6％(29/36) for complex CHD.The prevalence of neonatal CHD was 10.58％ (252/2382),including 241 with simple CHD and 11 complex ones. ConclusionsFour combined views of 2DE for prenatal screening is less sensitive in detecting simple CHD than complex CHD.Most of the complex CHD could be diagnosed by four combined views of 2DE before birth,but the misdiagnosis rate is high in simple CHD.The echocardiograms performed on newborns might make up for the lack.

BACKGROUND:At present,the most frequently agents used for neural induction of bone marrow stromal cells(BMSCs)in vitro are anti-oxidants,such as beta-mercaptoethanol and all trans-retinoids.The majorities of induction from BMSCs are neuron-like cells in these protocols;however,whether it has neuronal function or not should be further studied.OBJECTIVE:TO investigate the differentiated characteristics of inducing human BMSCs into neural cells in serum-free medium.DESIGN:Observational study.SETTING:Chaozhou Central Hospital.MATERIALS:The experiment was carried out in the Chaozhou Central Hospital from April 2004 to December 2005.Adult bone marrows were derived from femoral and tibial bone marrow of three patients with fracture.All patients provided the confirmed consent and were approved by the Ethics Committee of Chaozhou Central Hospital.DMEMIF12 medium(1:1),fetal bovine serum (FBS), glutamine, N2 supplements and B27 Supplements were from GIBCO/BRL Company;recombinant basic fibroblast growth factor(bFGF)and recombinant epidermal growth factor(EGF)from Sigma Company;monoclonal antibody for vimentin(1:100),monoclonal antibody for myelin basic protein(MBP) (1:100),monoclonal antibody for S1 00(1:1 00),monoclonal antibody for neuron specific enolase(NSE)(1:1 00),and monoclonal antibody for neurofilament 200(NF200)(1:1 00)from Beijing Zhongshan Company;monoclonal antibody for glial fibrillary acidic protein (GFAP)(1:200)and polyclonal antibody for nestin(1:100)from Boster Company(Wuhan);mouse monoclonal antibody for beta-tubulin 3(1:1 000)from Sigma Company;SP-9000 kits and quick AEC from Beijing Zhongshan Company; culture dishes and flasks from Coming Company.METHODS:BMSCs from human bone marrow were cultured in serum-containing medium.When the primary culture was established, BMSCs were transferred into serum-free medium containing N2 or B27 supplement with 20 μg/L basic fibroblast growth factor(bFGF)and epidermal growth factor(EGF),and cells were cultured in an incubator containing C02of 0.05 volume fraction at 37℃. Morphological changes of BMSCs in serum-free medium were observed under phase contrast microscope. And two days after culture. Expression of relative markers of BMSCs was detected withimmunocytochemistry.MAIN OUTCOME MEASURES:Morphological changes of BMSCs and expression of relative markers of nerve cells.RESULTS:A population of BMSCs could be isolated from adult human bone marrow,and they were processed to obtain a fibroblast-like population and were expanded as undifferentiated cells in culture for more than 10 passages.indicating their proliferative capacity.They could form spheroid state when they were sub-cultured in serum-free media supplemented with bFGF and EGF.these cells could express the markers for neural stem cells such as vimentin and nestin;they could expressed neuron specific enolase(NSE),beta-tubulin 3,TrkC and neurofilament 200(NF200)when they were plated on dishes with serum-containing medium; some cells exhibited the phenotypes for astrocytes.expressing gilal fibrillary acidic protein(GFAP)and S100 protein.CONCLUSION:The morphology,protein expression and differentiation ability of BMSCs in serum-free medium was similar to those of neural stem cells.The data support the hypothesis that adult bone marrow contains stem cells capable of differentiating into neural cells,the serum-free media make BMSCs overcome their mesenchymal commitment,showing the phenotypes for neural stem cells.

Objectives To observe the effect of Luoyutong capsule on neurological function following focal cerebral ischemia-reperfusion in rats and to preliminarily study the protective mechanism of Luoyutong capsule for focal cerebral ischemia-reperfusion in rats. Methods A rat model of middle cerebral artery occlusion (MCAO)was induced by the modified Longa method. After 1. 5 h of ischemia,reperfusion started. Ten male SD rats were selected as sham operation group,and forty male SD rats were randomly divided into 4 groups:Model (MCAO),Luoyutong moderate-dose (LYTM),Luoyutong high-dose (LYTH),and citicoline sodium (CS)groups (n=10 in each group). At day 3 and 7 after modeling,the neurological function of the rats was evaluated by using 12 neurological score and forelimb placing test. Western blotting was used to detect the expression levels of brain derived neurotrophic factor (BDNF),basic fibroblast growth factor (b-FGF),and phosphor/protein kinase (p-AKT/AKT)on the ischemic side of the rats and in the ipsilateral brain tissue at day 3 after modeling,as well as the expression level of Caspase-12 at day 7 after modeling in the ipsilateral brain tissue,and a comparison was performed among the groups. Results (1 )Neurological score:At day 3 after modeling,there was no significant difference between the 12 neurological score and the forelimb placing test score (all P>0. 05);At day 7 after modeling, there were obvious improvement in the LYTM,LYTH,and CS groups compared with model group (all P<0.05). (2)The results of western blot showed that①compare with the sham operation group,the expression levels of BDNF and b-FGF were reduced obviously (all P<0.05);compare with the MCAO group,the expression levels of the LYTM,LYTH and CS groups could be up-regulated,particularly in the LYTH group (P<0. 01);② compare with the sham operation group,the expression level of p-AKT/AKT in MCAO group was decreased obviously (P<0. 05);compare with the MCAO group,the expression levels of p-AKT/AKT of the LYTM,LYTH,and CS groups were increased,particularly in the LYTH and CS groups (all P<0. 05);③ compared with the sham operation group,the expression of cleavage Caspase-12 was increased obviously in the MCAO group (P<0. 05). Compared with the MCAO group,the expression levels of proCaspase-12 and cleavage Caspase-12 had a decreasing trend in the LYTM and LYTH groups,but there were no significant differences (all P >0. 05);the expression levels of proCaspase-12 and cleavage Caspase-12 in the CS group were obviously lower than those of the MCAO group (P<0. 05). Conclusion Luoyutong capsule may play a protective effect for focal cerebral ischemia-reperfusion injury in rats by promoting neural survival and regeneration,and this protective effect may be associated with the inhibition of neuronal apoptosis.

Objective To investigate the feasibility of laparoscopic D3 lymphadenectomy combined with pelvic autonomic nerve preservation for rectal cancer.Methods The clinical data of 34 patients with rectal cancer who had been admitted to OUr hospital from January 2007 to December 2007 were retrospectively analyzed.All patients underwent laparoscopic D3 lymphadenectomy combined with pelvic autonomic nerve preservation.The operation time,lymph node dissection,postoperative complications were assessed.The erectile function of male patients and the short-term outcome of the operation were evaluated.Results All patients were successfully operated on,and no conversion Was required.The mean operation time,blood loss,number of lymph node dissected,time of catheterization,mearl volume of residual urine and CEA were(265+46)minutes,(123±27)ml and 19±3,(5.5±1.6)days,(22.5±7.8)ml and(8.0±4.6)U/L,respectively.The incidence of postoperative complications and male sexual dysfunction were 9%(3/34)and 14%(3/21),respectively.No local recurrence or distal metastasis Was observed at the end of August 2008.Conclusions Laparoscopic D3 lymphadenectomy combined with pelvic autonomic nerve preservation for rectal cancer is relatively safe and feasible.

Objective To study the expression of VEGF-C and it's relationship with pathologic characteristics in rectal cancer.Method In this study 82 patients of mid to low rectal cancer underwent radieal resection from December 2007 to August 2008.Tumor tissues and lymph nodes were studied by immunohistochemical staining for VEGF-C expression in the tumor tissue and CK20 expression in D3 lymph nodes.Results were compared with the pathological results of mesenterium lymph nodes to explain the relationships between VEGF-C expressions and clinical pathologic characteristics.Data were analyzed with Chi square test.Result As for the expression of VEGF-C.significant differences were found between tumor stages(X~2=8.529,P<0.05)and between positive and negative lymph node metastasis(X~2=4.712,P<0.05).but there was no difference between that of mesenterium lymph node metastasis and D3 lymph node micrometastasis(X~2=0.017,P>0.05). Conclmion In rectal cancer,VEGF-C expression is correlated with tumor stage,type,metastasis and micrometastasis of lymph node.