Objective To investigate morphologic, structural and functional characteristics of cardiomyocytes from neonate rats and to set up a desirable technique for isolating and purifying the cardiomyocytes from neonate rats. Methods Using trypsin-digestion, mechanical separation, twice seedings and bromodeoxyuridine (BrdU)-treatment, the cardiomyocytes were isolated and purified from the hearts of neonate rats at 1-7 days after birth. Shapes and spontaneous pulsation of the cells were viewed. The cells were identified with cardiac isoform of tropnin T(cTnT) immunostaining. Ultrastructural features of the cells were examined with in situ transmission electron microscopy. Responses of the cells to adenine and isoprenaline were also examined. Results More than 95% cells isolated from the hearts of neonate rats are cardiomyocytes. The vital cells are more than 95%. Neonate rat cardiomyocytes include short columnar or rhabdoid cells and irregular cells. The most rhabdoid cells from the rats at 1-3 days after birth present the ultrastructural features of immature cardiomyocytes. The rhabdoid cells from the rats at 6-7 days after birth have some ultrastructural features of mature cardiomyocytes. Comparing with the cells at 1-3 days after birth, cTnT expression in the cells is slightly enhanced, the transverse striation was obvious. The irregular cells contain less bundles of myofilaments, the filaments are arranged irregularly. There are a few small cells which are in undifferentiated state. More than 80% cells show spontaneous pulsation at 72 hours after incubation. After treatment with adrenine and isoprenaline, the number of the cells with spontaneous pulsation increases and the intension of spontaneous pulsation is enhanced. The responses of the rhabdoid cells from the rats at 6-7 days after birth to adenine and isoprenaline are much stronger. Conclusion There are two kinds of neonate rat cardiomyocytes. They are different in ultrastructures, spontaneous pulsation and responses to adenine and isoprenaline. The cardiomyocytes from rats at 6-7 days after birth are suitable for experiments in vitro as mature cardiomyocyte. The method set up in this experiment is desirable for culture of neonate rat cardiomyocytes.

Objective To observe the effect of Tongsaimai Tablets on the leptomeningeal collateral circulation established after acute cerebral infarction by TCD.Methods 50 patients with acute cerebral infarction were randomly divided into control group (received conventional treatment) and treatment group (received Tongsainai Tablets three times a day,1.75g each time,on the basis of conventional treatment).The leptomeningeal collateral circulation of the two groups were observed after 6 months.Results The factors which may affect the leptomeningeal collateral circulation,such as age,the prevalence of hypertension and hyperlipidemia,original compensatory status of cerebral artery between the two groups had no statistically significant differences.The open rate of leptomeningeal collateral circulation in the treatment group was 64.00％,which was significantly higher than that of the control group(32.00％)(P =0.024).Conclusion Tongsaimai Tablets can promote the formation of leptomeningeal collateral circulation in patients with acute cerebral infarction.

Objective To observe the evolution process of cerebral blood flow spectrum in brain death by transcranial doppler ultrasound.Methods To detect the blood flow of bilateral middle cerebral artery in brain dead patients by transcranial doppler ultrasound.Results Twenty brain dead patients were collected in the study.The blood flow velocity of middle cerebral artery ranged s from 6 to 37cm/s,and the mean velocity was (12.13 ± 3.15)cm/s.There were three characteristic spectrums which include oscillatory wave (reverse wave in diastolic),nails wave (tip small wave in systolic),no blood flow sign,and they appeared in turn accoerding to worsen of the disease.The heartbeat irreversibly stopped within 8 days after the appearing of oscillatory wave,and ninty percent of the cases had the regular blood flow spectrums of oscillatory wave-nails wave-no blood flow sign.Conclusion The three regularly characteristic transcranial doppler ultrasound blood flow spectrums,which include oscillatory wave,nails wave and no blood flow sign could facilitate the diagnosis of brain death.

Objective To investigate the value of methylprednisolone pulse therapy on the acute treatment of multiple sclerosis.Methods 46 cases of the acute phase of patients with multiple sclerosis treatment in the hospital's department of neurology were chosen,who were divided in accordance with the principle of randomized controlled study group and the control group,each group of 23 patients.The study group was given methylprednisolone in the treatment and control group were given dexamethasone treatment.The clinical efficacy and complications of the treatment of patients were compared.Results The study group markedly in 14 cases,effective 54 cases,the total effective rate was 82.6％ ;the control group,10 casesmarkedly effective in 4 cases,9 cases ineffective,the total effective rate was 60.9％ ; compared the two sets of data decline before treatment,there was a statistically significant difference (x2 =5.236,P ＜ 0.05).After the treatment,the two groups of patients with EDSS scores compared with EDSS scores of the study group was significantly lower than that of the control group,the difference was statistically significant(t =3.135,P ＜ 0.05).Conclusion In acute phase of patients with multiple sclerosis,methytprednisolone pulse therapy can reduce the incidence of complications,and reduce the patient's nervous system damage,which can improve the quality of life in patients.

Objective To assess the predictive value of cerebral arteriosclerosis to acute cerebral infarction evaluated by transcranial doppler ultrasound .Methods 170 cases suspected cerebral arteriosclerosis consecutively enrolled were given transcranial doppler ultrasound and were followed up for 6 months,then we assessed the correla-tion between the levels of cerebral arteriosclerosis and acute cerebral infarction within 6 months.Results By Spearman rank correlation analysis ,there was positive correlation between the severity of cerebral arteriosclerosis de-tected by transcranial doppler ultrasound and acute cerebral infarction within 6 months,the correlation coefficient was 0.575(P<0.05).Conclusion Through the detection of transcranial doppler ultrasound ,the higher levels of cere-bral arteriosclerosis,the greater probability of acute cerebral infarction within 6 months.

Background Evidences indicated that oxidative stress damage is an essential pathological process in primary open angle glaucoma.Sulforaphane (SFN) can play an antioxidative stress role to many tissues and cells by activating Nrf2/ARE single pathway.However,whether SFN has a protective role to oxidative stress induced damage of trabecular meshwork cells is still unclear.Objective This study was to investigate the antioxidant effect of SFN against H2O2-induced oxidative damage in bovine trabecular meshwork cells.Methods Trabecular cells were isolated from fresh black bovine eyeballs and primarily cultured and passaged.The third generation of cells were incubated to 96-well dish at a density of 1 ×103/well for 24 hours and divided into 4 groups.The cells were incubated using 100 μl serum-free medium in the blank control group.Oxidative damage models were established by adding 100 μmol/L H2O2(100 μl) in medium in the H2O2 group.The cells were cultured with the medium containing 10 μmol/L SFN (100 μl) in the SFN group,and 100 μl H2O2 at the final concentration of 100 μmol/L was added in the SFN-treated cell medium in the SFN +H2O2 group.The cell vitality in various groups was assayed by using cell counting kit-8 (CCK-8).The apoptosis rate of the cells was detected by Annexin V-FITC/PI double-staining with flow cytometry.Results Cultured cells showed a spindle shape with uniform size,abundant cytoplasm,numberous pigmented particles and big nucleolus.The relative cell vitality reduced to (67.00± 1.27)％ and (80.00±6.25)％ in the H2O2 group and SFN+H2O2 group in comparison with 100％ in the blank control group,and the cell vitality in the SFN+ H2O2 group was lower than that in the SFN group but higher than that in the H2 O2 group (both at P＜0.01).The mean apoptosis rate was (11.33 ±0.77) ％,(32.31 ± 1.03) ％,(10.44 ±0.68) ％ and (17.68 ±0.21) ％ in the blank control group,H2 O2 group,SFN group and SFN+H2O2 group,respectively,showing a significant difference among the groups (F=539.96,P＜0.01),and the apoptosis rate in the SFN+H2O2 group was significantly lower than that in the H2O2 group but higher than that in the blank control group and SFN group (all at P＜0.01).Conclusions SFN can improve the antioxidative stress ability of trabecular meshwork cells and alleviate the damage induced by oxidative stress.

Objective To investigate the distrbution of actin and the level of Ca 2+ in macrophages activated with LPS and IFN\|? and study effects of reorganization of actin and the level of Ca 2+ on migration and phagocytosis of macrophages. Methods Rat macrophages were activated with LPS and INF\|?,actin and intracellular free Ca 2+ were labeled with phallacidin and Fura\|2,the distribution of actin and the level of Ca 2+ were observed and analyzed with Ca 2+ image analysis system and confocal laser scanning microscope. Results The migrating cell and the phagocytizing cells increased in the activated macrophages.F\|actin and the level of Ca 2+ of the activated macrophages increased.There was a front and a back in the migrating cell.Lamellipodia and filopodia protruded from the front.F\|actin of the lamellipodium and filopodium was rich.There were stress fibers of F\|actin at the bottom of the migrating cells.In the migrating macrophages,the level fo Ca 2+ in the back was high than that in the front.When the migrating cell turned their growth direction,the level of Ca 2+ at the curve increased greatly,especially in its lateral region.When macrophage phagocytized the dead lymphocyte,Ca 2+ in the phagocytizing region increased. Conclusion When macrophages are activated with LPS and IFN\|?,migration and phagocytosis increase greatly.Particular morphologic changes in the distribution of F\|actin and the level of intracellular free Ca 2+ occur. [

Objective In order to investigate effects of extracellular matrix on lymphangiogenesis. Methods Endothelial cells of canine thoracic duct were incubated in type I collagen gel and on Matrigel basement membrane matrix gel.Morphologic changes of endothelial cells and tube formation were observed under phase contrast microscope,structures of lymphaticlike channels were observed with transmission electron microscope,tube formation was quantified by measuring the length and area of tubes with image analyzer. Results Lymphatic endothelial cells on type Ⅰ collagen gel grew to confluent monolayers,no tubes were found.The cells underwent morphologic changes and reorganized into lymphaticlike tubes when the cells were covered with the top gel layer.Tubes of heparin\|treated dishes were more than that of the control dishes,tubes of dishes treated with heparin and bFGF together were more than that of dishes treated with heparin or bFGF alone.Endothelial cells on Matrigel basement membrane matrix gel migrated into the upper part of the gel and formed tubes.The tubes were thin,the cells became long.The tubes were in traction obviously.In ultrathin sections,tubes formed by lymphatic endothelial cells represented morphologic characteristics of lymphatic capillary.The lymphaticlike tubes contained matrix.Conclusion\ Endothelial cells of canine thoracic duct reorganized into lymphaticlike channels in type Ⅰ collagen gel and Matrigel basement membrane matrix gel.Extracellular matrix plays important roles in morphological changes and migration of lymphatic endothelial clells and lymphangiogenesis.\;[

Objective To compare features in expression of adhesion molecules of immunoglobin super family on lymphatic,vascular and microvascular endothelial cells and explore implications in expression of the adhesion molecules of immunoglobin super family on lymphatic endothelial cells. Methods Endothelial cells were isolated from canine thoracic ducts,common carotid arteries,internal jugular veins and pulmonary microvessels.Expression of PECAM-1,ICAM-1,ICAM-3,VCAM-1 and CD44 on all kinds of endothelial cells were examined by immunofluorescence staining and viewed using a fluorescence microscope and a confocal laser scanning microscope.Expression strength of the adhesion molecules was analyzed with an image analyzer. Results The arterial,venous and microvascular endothelial cells expressed PECAM-1,ICAM-1,ICAM-3,VCAM-1 and CD44.Expression of ICAM-1 and ICAM-3 on the cells was weak.Expression of VCAM-1 on the arterial and microvascular endothelial cells was stronger than that on venous endothelial cells.The lymphatic endothelial cells expressed PECAM-1,ICAM-1,ICAM-3 and CD44,expression of VCAM-1 was not observed.Expression of ICAM-3 and CD44 on the cells was stronger than that on vascular and microvascular endothelial cells.Conclusion\ In comparison with the arterial,venous,microvascular endothelial cells,lymphatic endothelial cells do not express VCAM-1,expression of ICAM-3 and CD44 is stronger.This is helpful to explain mechanisms of adhesion of lymphocytes or tumor cells to the lymphatic endothelium and lymphangiogenesis.

Tumor is still an unresolved health problem for human.The main obstacle for tumor cure is that precise diagnosis of tumor at the early stage can't be realized,which leads to the mortality increased year by year due to missing the best time of the treatment.Furthermore,lack of targeted ability and serious side effects of the medicine used for tumor treatment also limit the clinical application.Therefore,it is urgent to design and develop a novel targeted drug delivery vehicle.Recent studies show that targeted nanomaterials are superior on tumor-specific diagnosis and target therapy.This review will highlight the current advances in the targeted nanomaterials effect on tumor diagnosis and treatment in recent years and prospect the foreground of future development.