Objective To investigate the chemical constituents in the leaves of Dracaena cochinchinensis.Methods The compounds were separated with column chromatography and their chemical structures were identified by physicochemical and spectral method,respectively.Results Thirteen compounds were isolated from the plant. They were identified as isorhamnerin(Ⅰ),quercetin(Ⅱ),25(R)-spirostane-5-en-3?-ol(Ⅲ),gracillin(Ⅳ),25(R)-spirostane-5-en-3?,14?-diol(Ⅴ),25(R)-spirostane-5-en-3?,14?-diol-3-O-?-D-glucopyranoside(Ⅵ),25(R)-spirostane-5-en-3?,14?-diol-3-O-?-L-rhamnopyranosy(1→4)-?-D-glucopyranoside(Ⅶ),25(R)-spirostane-14?-hydroxy-4-en-3-one(Ⅷ),7?-hydroxysistosterol-3-O-?-D-glucopyranoside(Ⅸ),?-stigmasterol(Ⅹ),stigmasterol-3-O-?-D-glucopyranoside(Ⅺ),daucosterol (Ⅹ Ⅱ),and methyl ?D-glucopyranoside(Ⅹ Ⅲ).Conclusion Spirostane-type steroids are the major constituents in the leaves of D.cochinchinensis.Compounds Ⅰ-Ⅲ,Ⅴ,Ⅵ,Ⅷ,and Ⅸ are isolated from this plant for the first time.

Objective To investigate the relationship between serum CC10 protein and lung injury induced by mechanical ventilation.Methods Forty healthy Wistar rats of both sexes weighing 200-250 g were randomly divided into 5 groups(n=8 each):group Ⅰ control;group Ⅱ mild lung injury[VT=7 ml/kg,duration of mechanical ventilation(t):2 h];group Ⅲ moderate lung injury(VT=7 ml/kg,t=4 h);group Ⅳ severe lung injury(VT=40 ml/kg,t=2 h);group Ⅴ extremely severe lung injury(VT=40 ml/kg,t=4 h).The animals were anesthetized with intraperitoneal 10% chloral hyrdrate 3.5 ml/kg and tracheostomized.Group Ⅰ received no mechanical ventilation.The animals in group Ⅱ-Ⅴ were mechanically ventilated with air(FiO2=21%,RR=40 bpm,I:E=1:2).The animals were sacrificed at the end of mechanical ventilation.The lungs were immediately removed for microscopic examination and determination of W/D lung weight ratio.The left lung was lavaged.The CC10 protein level in broncho-alveolar lavage fluid(BALF)and serum were determined by Western blotting.The Clara cells in the bronchiolar epithelium were examined by immno-histochemistry.Results The level of CC10 protein in BALF was significantly lower in group Ⅳ and Ⅴ while the serum CC10 protein level was significantly higher in group Ⅱ-Ⅴ than in group Ⅰ.The serum CC10 protein level was positively correlated while the CC10 protein level in BALF was negatively correlated with the severity of lung injury and W/D ratio.Conclusion The serum CC10 protein is closely related to the severity of lung injury induced by mechanical ventilation.

Objective To explore the effect of nitric oxide(NO) on pulmonary tumor necrosis factor?(TNF?) in murine acute necrotizing pancreatitis(ANP). Method Ninety-six SD rats were randomized into four groups:normal control group, ANP group, L-Arginine(L-Arg) pretreatment group and L-NAME pretreatment group (n=24 for each group). The protein content of bronchoalveolar lavage fluids(BALF), the myeloperoxidase(MPO) of lung tissue and TNF? produced by alveolar macrophages were evaluated. The expression of TNF? mRNA was measured. ResultMPO and protein of BALF reached a peak level at 12nd hr (10.78?0.58U/g for MPO and 2011?106?g/ml for protein content respectively).TNF? peaked on the sixth hour(1624?149)pg/ml. The expression of AM TNF?mRNA also peaked on the sixth hour (1.127?0.069) along with an increase of TNF? mRNA. A similar tendency was seen in L-Arg and L-NAME pretreatment groups, with changes being statistically different in the three groups when compared with that of normal control group(P

BackgroundCorneal graft reject is a major cause of corneal transplantation failure.Although many immune-suppressing drugs have been utilized to reduce the reject response,their adverse effects on organ and tissue are still insoluble.The tolerance induction of natural killer T (NKT) cells is currently under investigation.However,the study on the application of NKT cells in high risk corneal transplantation is seldom.ObjectiveThe present study was to explore the effects of α-GalCer-activated NKT cella on allografts survival after high-risk corneal transplantation surgery via retro-bubar injection.Methods The lymphocytes were picked up from the spleen of SPF Lewis rats and cultured in RPMI 1640 medium with 100 mg/L α-GalCer.After one week,NKT cells were sorted by the FACSVantage system as CD161+ TCR-α+ cell from the lymphocytes with the cell densities 5×106/ml.Ten SPF Fisher344 rats were used to prepare the donor corneas,and 20 Lewis rats served as recipients.The high risk corneal transplantation models were created by corneal suturing in 20 recipient rats.Penetrating keratoplasty (PKP) was performed in the model rats.0.1 ml NKT cells or the same volume of normal saline solution were retro-bubarly injected at the end of surgery respectively.The corneal allografts were observed and scored based on Holland criteria at the three-day interval under the slit lamp for 30 days.Two weeks after surgery,three rats from each group were sacrificed by excessive anesthesia method and the eyeballs were obtained for histopathological examination.The inflammatory cell infiltration ( CD4+ and CD8+ ) in grafts was evaluated by immunochemistry and flow cytometry.The use of the animals complied with the Statement of ARVO.ResultsThe mean survival time of the allografts was (7.90± 1.37) days in normal saline solution group and (14.70± 1.49) days in NKT cell group,showing a statistically significant difference between the two groups ( t =10.61,P =0.00 ).Two weeks after surgery,all the allografts showed the severe opacity with lots of new blood vessels and edema in normal saline solution group.However,the corneal grafts were clear in NKT cell group.Abundant CD4+ and CD8+T lymphocytes were seen in the allografts in normal saline solution group,but the inflammatory cells were obviously less in NKT cell group.The percentage of NKT cells in the spleen was (5.67±0.25)％ in NKT cell group and ( 1.21±0.19)％ in normal saline solution group ( t =8.43,P =0.00 ).Conclusionsα-GalCer-activated NKT cells can prolong the survival time of allografts in high-risk corneal transplantation.Retro-bubar injection of α-GalCer-activated NKT cells probably is a new approach to the prevention of the rejection of corneal transplantation.

5-(4-Hydroxyphenyl)-3H-1,2-dithiole-3-thione (ADT-OH) is a slowly releasing H2S donor with some neuroprotective effect.In order to study the structure-activity relationships,seventeen compounds (Y1-Y17) were synthesized by modification of ADT-OH at the aromatic ring position,and their structures were confirmed by 1H NMR,13C NMR and HR-MS.Among them,6 compounds (Y4,Y13-Y17) were novel compounds.Their effects had been evaluated on HT-22 hippocampal neuron cells damaged by glutamate with MTF method.The pharmacological results demonstrated that all the Y compounds had potent neuroprotection at most of the tested concentrations (1-100 μmol/L).Compounds Y1-Y9 and Y11 significantly improved the survival rates of the damaged cells at 1-100 μmol/L (P ＜0.01).Specially,compounds Y1,Y4,Y6-Y9,Yu are more potent than their parent compound ADT-OH at concentration of 1-10 μmol/L,which is worthy of further study.

Aim To investigate the effect of TRPV4 on hepatic fibrosis of rats . Methods Liver fibrosis model of rats was induced by 50% CCl4 twice a week for 12 weeks. HE and Masson staining were used to evaluate the degree of hepatic fibrosis, and the levels of α-smooth muscle actin(α-SMA) and TRPV4 were detec-ted in fibrotic liver tissue by Western blot. HSC-T6 cells were activated by transforming growth factor β1 (TGF-β1),and the protein levels of α-SMA, TRPV4 were detected by Western blot. After using Ru and transfected with TRPV4-siRNA, HSC-T6 was stimula-ted with TGF-β1, the levels of α-SMA, TRPV4 and phosphorylation level of Akt were determined by West-ern blot. Results TRPV4 was highly expressed in model liver tissues and in activated HSC-T6 induced by TGF-β1 . The levels of α-SMA and phosphorylation of Akt decreased in TGF-β1-induced HSC, used with Ru or transfected with TRPV4-siRNA. Conclusions The expression of TRPV4 increases in fibrotic livers and ac-tivated hepatic stellate cells. Knockdown of TRPV4 can suppress the activation of hepatic stellate cells in-duced by TGF-β1 , and decrease the phosphorylation levels of Akt.

Objective To explore the in-vivo hemodynamic and hemolysis effect of a newly designed axial continuousflow ventricular assist device(VAD) in swine.Methods Under general anesthesia,each of 5 swine [weight (40.0 ± 5.2)kg] was implanted with the axial continuous-flow VAD into the apex of left heart ventricle,and the outflow graft was anastomosised to descending aorta.Results All of the axial continuous-flow VAD were implanted successfully with post-operative survival rate 100％.All 5 animals survived over one week.There was a positive correlation between pump speed and assistance effect.The mean left ventricular systolic pressure was (131.6 ± 28.0) mmHg(1 mmHg =0.133 kPa).While the axial continuous-flow VAD was working,left ventricular end diastolic pressure decreased,along with mean intraventricular pressure declined.Peripheral hemodynamics was stable and peripheral blood pressure was not remarkably different from the pressure preoperation.Daily urine volume was in normal range within 1 week post operation.Free hemoglobin in plasma was slightly elevated on the surgery day,and gradually dropped to normal level within 1 week.International Normalized Ratio(INR) was maintained between 2.0-2.5 with oral adminiatration of warfarin of 3 mg/day.There was no thrombosis existing in VAD at autopsy.Conclusion The application of the axial pump with hydrodynamic-magnetically levitated impeller in animal experiment can provide stable hemodynamics,advanced heart unloaded effect,favorable peripheral perfusion,and blood compatibility is satisfactory.

BACKGROUND:Micro-fracture surgery method is simple, easy to operate, which is an effective way to treat articular cartilage defects, but there are stil some problems such as regenerated fibrocartilage and regenerated cartilage degradation. Scholars have focused on the use of various methods to improve the micro-fracture effect on repairing cartilage defects.
OBJECTIVE:To explore the effects of micro-fracture enhanced by autologous bone marrow mesenchymal stem cells extracellular matrix (aBMSC-dECM) scaffold for treating cartilage defects in minipig models.
METHODS:Bone marrow was extracted from the minipigs and bone marrow mesenchymal stem cells were obtained. aBMSC-dECM membranes were col ected. Cross-linking and freeze-drying technology were used to make the three-dimensional porous aBMSC-dECM scaffold. Ful thickness cartilage defects, 2 mm in depth and 6 mm in diameter, were created on the femoral condyles and trochlea grooves of the two knees of the minipigs. The right knees were treated with micro-fracture as control and the left were treated with micro-fracture enhanced by aBMSC-dECM scaffold. Six months later, histological examination and Wakitani score were used to evaluate the cartilage regeneration, and glycosaminoglycans and DNA contents in the regenerative tissue were determined.
RESULTS AND CONCLUSION:After 6 months, the tissue treated by micro-fracture enhanced by aBMSC-dECM scaffold got better surface and integrated with the surrounding cartilage. Safranin O and fast green staining and Masson staining showed that the regenerated cartilage of the left knee, with abundant matrix and dense bone trabeculae, was better than that of the right. Wakitani score of the left knee was higher than that of the right. Glycosaminoglycans content of the left knee was much more than that of the right, while the DNA content was lower in the left knee than the right knee. Better results were observed in the left knee undergoing micro-fracture enhanced by aBMSC-dECM scaffold, and improvements in the femoral condyles and trochlea grooves showed no differences.

Objective To investigate the influence of IVIG on immunologic function and cytokines levels in children with EV71 infection associated high-risk pulmonary hemorrhage. Methods According to the inclu-sion criteria , 64 children were enrolled and randomly divided into two groups: 39 cases in the IVIG treatment group and 25 cases in the general treatment group. The alternations of blood and immune cytokine markers before and after treatment were detected in the patients. Results (1) Before treatment, the peripheral blood T cells, TH and B cells in the IVIG group were higher than those in the general group , but the peripheral blood IgA was lower than that in the general group(P < 0.05); (2) In the IVIG group, the NK cells and IgG increased and T cells, TH cell and B cells significantly decreased after receiving IVIG therapy (P < 0.05); (3)No significant difference was observed in the levels of IL-6 and IL-10 in the two treatment groups before treatment. However , after IVIG treatment, IL-6 and IFN-γ levels reduced, IL-10 level increased (P < 0.05). After the general treat-ment, no significant difference was found in levels of IL-6,IL-10 and IFN-γ in peripheral blood (P > 0.05). Conclusion Disorders of cellular immunity and humoral immunity appeared in children with EV71 infection-re-lated high-risk pulmonary hemorrhage. It has clinical value to use IVIG timely to regulate the immune disorder.

Objective:To explore the association of physical activities,sedentary behaviors with over-weight/obesity in Chinese children aged 9 -1 1 years,analyze the gender difference and heterogeneity of the association across different provinces,and provide evidence for prevention and control of overweight and obesity.Methods:Using the data of 40 692 children aged 9 -1 1 years from Chinese National Survey on Students’Constitution and Health in 201 0,the height and weight were measured and the time spent in physical activities,screen behaviors and homework were collected.We analyzed the associations among physical activities,sedentary behaviors and overweight/obesity in boys and girls separately,and assessed the heterogeneity of associations across different provinces with Meta-analysis.Results:The prevalence of overweight/obesity of the boys (27.1 %)was higher than that of the girls (1 2.9%),compared with the rural students (1 5.6%),the situation of overweight/obesity of the urban students (23.8%)was more serious,the prevalences of overweight/obesity were separately 22.6%,1 9.2% and 1 7.5% among developed,middle-developed and undeveloped social economic status subgroups.The boys with physical activities no more than 1 hour per day were more likely to be overweight/obese,with the OR of 1 .09 (95%CI:1 .02,1 .1 7).The girls with screen time more than 1 hour per day or homework time more than 2 hours per day were more likely to be overweight/obese,with the ORs of 1 .1 3 (95%CI:1 .02, 1 .26)and 1 .1 8 (95%CI:1 .03,1 .35)respectively.The high level of sedentary behaviors (more than 1 35min per day)was associated with overweight/obese in the girls,and the OR was 1 .1 9 (95%CI:1 .08,1 .33).There was no significant heterogeneity of effect values between physical activities and over-weight/obesity in the boys and between sedentary behaviors and overweight/obesity in the girls across the different provinces (P >0.05).Conclusion:The associations among physical activities,sedentary be-haviors and overweight/obesity existed in Chinese children aged 9 -1 1 years,which were different in boys and girls.There was no statistically significant heterogeneity across different provinces.The strate-gies to prevent and control overweight/obesity in primary school students should focus on gender-specific needs.