Object To study the protective effect of crocetin on myocardial ischemia induced by isoproterenol (ISO). Methods Myocardial ischemia was induced by subcutaneous injection of ISO (30 mg/kg for two days, once a day). The experimental animals were randomly divided into the normal group, ISO group, positive control group and three crocetin groups (25, 50, 100 mg/kg). Cardiac indexes were examined. The level of LDH, CK, MDA in serum were measured. The content of MDA, GSH-Px and the ATPase activity heart and mitochondria were assayed by colorimetric analysis. The histopathological change of myocardia was investigated by HE staining. Results Compared with the model group, crocetin can significantly reduce the cardiac indexes, obviously decrease the level of MDA, LDH and CK in the serum. Crocetin can significantly increase the activities of Na +, K +-ATPase; Ca 2+ , Mg 2+ -APTase; GSH-Px. The histopathological changes confirmed the protective effects of crocetin on the myocardial injury. Conclusion Crocetin could alleviate the acute myocardial ischemia induced by ISO in rats.

Objective To detect the Telomerase inhibitor-antisense phosphorothiate oligonucleotide (asONS) effect on telomerase activity of human bladder tumor cell line BIU-87.Methods BIU-87 cells and cell extracts are cultured with asONS,non-asONS and control fluid at different concentrations and times respectively.Telomerase activity was assayed by TRAP PCR-ELASA.Results AsONS can inhibit the activity of BIU-87 cells ,whereas non-asONS and control group not, and have significant difference(P

Objective To evaluate the long term results of coral bone used in anterior cervical interbody fusion. Methods The 180 patients (126 males and 54 females) who had undergone anterior interbody fusion in our hospital were followed up for 6 years. Their long-term imageological data, including changes in interbody height, interbody angle and range of movement, were analyzed and compared. Results In all the cases a solid fusion was achieved between coral reef and vertebra. The fusion was evaluated as successful in all the 180 patients. Conclusion The coral bone spacer provides the same solid fusion as autografting with iliac crest does.

Objective To investigate the mechanisms of tumor necrosis factor-related apoptosis inducing ligand (TRAIL) combined with Triptolide in inducing the apoptosis of pancreatic cancer cells.Methods (1) The pancreatic cancer cells (MiaPaca-2 cells) were divided into 4 groups:blank control group (no drugs were added),TRAIL + Triptolide-group (only TRAIL was added),TRAIL-Triptolide + group (only Triptolide was added) and TRAIL+ Triptolide+ group (TRAIL and Triptolide were added).The vitality of cells in all the 4 groups was assessed by CCK-8.The expressions of poly ADP-ribose polymerase (PARP),cysteinyl aspartate specific proteinase-3 (Caspase-3) and Caspase-8 were detected by Western blot.The vitality of cells was detected by CCK-8 and the vitality of Caspase-8 was detected by Caspase-Glo assays after adding Z-IETD-FMK,a specific inhibitor of Caspase-8.The expressions of myeloid cell leukemia-1 (Mcl-1),Bcl-xL and Bcl-2 were detected by Western blot.(2) The MiaPaca-2 cells were divided into 8 groups:①TRAIL-Mcl-1 siRNA-group (no TRAIL was added and Mcl-1 siRNA cells were not transfected),TRAIL+ Mcl-1 siRNA-group (TRAIL was added and Mcl-1 siRNA cells were not transfected),TRAIL-Mcl-1 siRNA + group (TRAIL was not added and Mcl-1 siRNA cells were transfected)and TRAIL+ Mcl-1 siRNA+ group (TRAIL was added and Mcl-1 siRNA cells were transfected).②TRAIL-Bcl-xL siRNA-group (TRAIL was not added and Bcl-xL siRNA was not transfected),TRAIL+ Bcl-xL siRNA-group (TRAIL was added and Bcl-xL siRNA was not transfected),TRAIL-Bcl-xL siRNA + group (TRAIL was not added and Bcl-xL siRNA was transfected) and TRAIL+ Bcl-xL siRNA+ group (TRAIL was added and Bcl-xL siRNA was transfected).The vitality of the cells in all the groups was detected by CCK-8.The expressions of Caspase-3 and Caspase-8 protein were detected by Western blot.The measurement data with normal distribution were presented as (x) ± s.The comparison among groups was done by ANOVA,and the pairwise comparison was done by LSD-t test.Results (1) The vitalities of MiaPaca-2 cells in the blank control group,TRAIL + Triptolide-group,TRAIL-Triptolide + group and TRAIL + Triptolide + group were 100.0％ ± 1.1％,81.2％ ± 2.3％,78.6％ ± 3.6％and 40.1％ ± 2.5 ％,and the relative expressions of PARP protein were 0.510 ± 0.028,0.720 ±0.072,1.250 ±0.023 and 2.560 ± 0.220,the relative expressions of Caspase-3 were 0.080 ± 0.004,0.080 ± 0.003,0.110 ±0.005 and 2.720 ± 0.003,and the relative expressions of Caspase-8 were 0.070 ± 0.003,0.080 ± 0.005,0.120 ±0.003 and 0.990 ± 0.006,with significant differences among the 4 groups (F =203.607,1 457.785,332 421.900,35 437.218,P ＜ 0.05).The vitality of M iaPaca-2 cells in the TRAIL + Triptolide + group was significantly different from those in the blank control group,the TRAIL + Triptolide-group and the TRAIL-Triptolide + group (t =34.583,355.936,36.271,P ＜ 0.05).The relative expression of PARP protein of MiaPaca-2 cells in the TRAIL+ Triptolide + group was significantly different from those in the blank control group,TRAIL+ Triptolidegroup and TRAIL-Triptolide + group (t =591.784,63.739,2 268.987,P ＜ 0.05).The relative expression of Caspase-3 protein of the MiaPaca-2 cells in the TRAIL + Triptolide + group was significantly different from those in the blank control group,the TRAIL + Triptolide-group and theTRAIL-Triptolide + group (t =3 266.153,9 145.228,1 738.713,P ＜0.05).The relative expression of Caspase-8 protein of the MiaPaca-2 cells in the TRAIL+ Triptolide +group was significantly different from those in the blank control group,the TRAIL+ Triptolide-group and the TRAIL-Triptolide + group (t =663.953,l 432.878,327.584,P ＜ 0.05).The vitality of caspase-8 in the TRAIL+ Triptolide+ group was 711.0％ ± 5.1％ before adding Z-IETD-FMK,and then the vitality of MiaPaca-2 cells and caspase-8 changed to 70.0％ ± 4.8％ and 73.0％ ± 2.4％,with significant differences (t =17.956,55.027,P ＜ 0.05).The relative expressions of Mcl-1 protein in the blank control group,the TRAIL + Triptolidegroup,the TRAIL Triptolide + group and the TRAIL + Triptolide + group were 1.68 ± 0.22,2.08 ± 0.11,0.73 ±0.15 and 0.58 ± 0.18,the relative expressions of Bcl-xL protein were 0.65 ± 0.03,0.47 ± 0.03,0.32 ± 0.03and 0.26 ±0.05,the relative expressions of Bcl-2 protein were 0.65 ± 0.03,0.67 ± 0.03,0.62 ± 0.05 and 0.67 ± 0.03,with significant difference among the 4 groups (F =55.178,88.683,3.411,P ＜ 0.05).The relative expressions of Mcl-1 protein of the MiaPaca-2 cells in the TRAIL-Triptolide + group and the TRAIL+ Triptolide +group were significantly different from those of the blank control group (t =23.506,47.631,P ＜ 0.05) and the TRAIL + Triptolide-group (t =58.457,37.115,P ＜ 0.05).The relative expressions of Bcl-xL protein of the MiaPaca-2 cells in the TRAIL-Triptolide + group and the TRAIL + Triptolide + group were significantly different from those of the blank control group (t =38.105,42.219,P ＜ 0.05) and the TRAIL + Triptolide-group (t =32.476,15.814,P ＜ 0.05).The relative expressions of Bcl-2 protein in the TRAIL-Triptolide + group and the TRAIL+ Triptolide + group were not significantly different from those of the blank control group (t =4.724,1.732,P > 0.05) and the TRAIL + Triptolide-group (t =3.464,0.000,P ＞ 0.05).(2) The vitalities of MiaPaca-2 cells of the TRAIL-Mcl-1 siRNA-group,TRAIL + Mcl-1 siRNA-group,the TRAIL-Mcl-1 siRNA + group and the TRAIL + Mcl-1 siRNA + group were 100.0％ ± 2.2％,79.3％ ± 1.8％,71.2％ ± 3.2％ and 37.3％ ± 5.4％,the relative expressions of Caspase-8 protein were 0.100 ± 0.003,0.100 ± 0.005,0.100 ± 0.003 and 0.350 ±0.005,and the relative expressions of Caspase-3 protein were 0.020 ± 0.003,0.060 ± 0.003,0.020 ± 0.003 and 0.590 ±0.004,with significant differences among the 4 groups (F =136.681,2 717.391,44 471.429,P ＜0.05).The vitality of MiaPaca-2 cells of the TRAIL + Mcl-1 siRNA + group was significantly different from those in the TRAIL-Mcl-1 siRNA-group,the TRAIL + Mcl-1 siRNA-group and the TRAIL-Mcl-1 siRNA + group (t =33.937,20.207,26.689,P ＜ 0.05).The relative expression of Caspase-8 protein of the TRAIL + Mcl-1 siRNA +group was significantly different from those in the TRAIL-Mcl-1 siRNA-group,the TRAIL + Mcl-1 siRNA-group and the TRAIL-Mcl-1 siRNA + group (t =216.506,433.013,144.338,P ＜ 0.05).The relative expression of Caspase-3 protein of the TRAIL + Mcl-1 siRNA + group was significantly different from those in the TRAIL-Mcl-1 siRNA-group,the TRAIL + Mcl-1 siRNA-group and the TRAIL-Mcl-1 siRNA + group (t =329.09,458.993,987.269,P ＜0.05).The vitalities of MiaPaca-2 cells of the TRAIL-Bcl-xL siRNA-group,the TRAIL+ Bcl-xL siRNA-group,the TRAIL-Bcl-xL siRNA + group and the TRAIL+ Bcl-xL siRNA + group were 100.0％ ± 2.3％,87.2％ ± 4.1％,74.1 ± 3.7％ and 56.3％ ± 5.4％,and the relative expressions of Caspase-3 protein were 0.060 ±0.004,0.070 ± 0.003,0.060 ± 0.004 and 0.390 ± 0.003,with significant differences among the 4 groups (F =70.074,4 643.478,P ＜ 0.05).The vitality of MiaPaca-2 cells of the TRAIL + Bcl-xL siRNA + group was significantly different from those in the TRAIL-Bcl-xL siRNA-group,the TRAIL+ Bcl-xL siRNA-group and the TRAIL-Bcl-xL siRNA + group (t =24.416,41.170,18.136,P ＜ 0.05).The relative expression of Caspase-3 protein of the TRAIL + Bcl-xL siRNA + group was significantly different from those in the TRAIL-Mcl-1 siRNA-group,the TRAIL + Mcl-1 siRNA-group and the TRAIL-Mcl-1 siRNA + group (t =285.788,554.256,190.526,P ＜ 0.05).Conclusion Triptolide could induce the apoptosis of MiaPaca-2 cells by inhibiting the expressions of Mcl-1 and Bcl-xL,sensitizing TRAIL and activating Caspase-8 and Caspase-3.

Objective To explore the effects of glycosylation in E2 and E1 protein on specific cell fusion in rubella virus(RV)strain JR23.Methods Site-directed mutagenesis was used tO obtain mutants containing new enzyme sites on the E2 and E1 gene of RV JR23.All the mutants and wild type proteins were expressed in BHK21 cells and treated with acid medium to induce specific cell fusion.The fusion functions were assayed with Giemsa staining and reporter gene method for qualitative and quantitative analysis,respectively.Expression efficieneies of mutant proteins on cell surface were quantified with fluorescence-activated cell sorter(FACS).Hemadsorption assays were performed to detect binding activity of mutant proteins qualitatively and quantitatively.Results Mutant proteins E2 N53G,S73I and S131V had 62.73%,66.66%and 55.12% of fusion activities,and E1 T78A,T179A and T211A had 66.93%,87.33%and 90.18%of fusion activities,respectively,as compared with the wild type protein.The FACS indicated that the expression efficiencies of all the mutant proteins except E2 S131V were lower than that of the wild type protein.Hemadsorption assays demonstrated that binding abilities of E2 S73I and E1 T78A decreased slightly,but that of the other four mutant proteins Wns almost same as the wild type protein. Conclusion Glycosylation on E2 N53,N71,N129 and E1 N76 were important for the specific cell fusion,but E1 N177 and N209 were almost not.

Objective To investigate the prevalence of human immunodeficiency virus (HIV) infection and its characteristics among pregnant women in Hunan.Methods Data from information system of prevention of mother-to-child transmission of HIV management in Hunan 2011-2015 was analyzed in the study (3 + 1 mode by year statistics).Results The total HIV-positive infection rate was 0.19‰ among pregnant women from 2011 to 2015 in Hunan.The rate of HIV infection showed upward trend by years (P ＜ 0.05).The proportion of diagnosis of HIV positive cases intrapartum was 44.66％,showed declining trend by years (P ＜ 0.05).The 786 cases of HIV positive pregnant women were mainly the Han's,the age distribution of 20 to 35 years old,90.21％ of them were farmers or unemployed.A percentage (76.84％) of them had junior high school education level or lower 37.91％ of them were found in delivery or postpartum.A percentage (36.51％) of them accepted service in delivery or postpartum.A percentage (45.67％) of them was infected through sexual contact,46.82％ of them were infected by unknown ways.Conclusions The rate of HIV infection among pregnant women was increased by years in Hunan.It is suggested to strengthen health education among high-risk groups and high incidence areas,improve detection rate of early pregnant women,implement the Prevention of Mother to Child Transmission (PMTCT) measures to reduce the rate of mother to child transmission of Acquired Immune Deficiency Syndrome (AIDS).

AIM:To study the effect of Danhong Zhiganching capsules on the experimental fatty liver in rats.METHODS: Male SD rats were treated by high fat diet for 6-week after being treated by a low dose of CCl_4 to induce fatty liver model.Then,drugs were given by oral to the rats,the contents of triglyeride,total cholesterol in serum and liver tissue,the contents of superoxide dismutase and malondialdehyde in serum and the contents of free fatty acids in liver tissue acted as indexes to determine the effect of Danhong Zhiganching capsules.RESULTS: The contents of triglyeride and total cholesterol in serum and liver tissue of rats were significantly reduced by Danhong Zhiganching capsules,and so it is with the contents of free fatty acids in liver tissue(P

objective :Tostudytherelationshipbetweenthenumberofcellclonewithcalcificationpotencyand theapplicationtimeofe PTFEmembraneofguidedtissueregeneration .Methods :e PTFEwasappliedinthe periodontaldefectsof 2 4teethin 4dogs .2 ,4and 8weeksafteroperationthetissuebetweenperiodontalmem braneandrootsurfacewasscaledandculturedin? MEM ,afterprimaryculture ,thecellswereclonedandcul turedinthepresenceofdexamethasoneformineralization .Results ::2 ,4and 8weeksafterapplicationofe PTFE 96,71and 10 4cellcloneswereobtainedandthemineralizationratio(% )ofthecloneswas 72 ,69and 3 9 (2or 4weeksvs 8weeksP 0 .0 5 ) ,respectively .Conclusion :Theresultssug gestthate PTFEmembraneshouldbetakenoutinnolessthan 6weeksafterappliedinperiodontaldeffects .

Objective To study the blood supply of the flap accompanying vessels of the cutaneous nerves in the lower leg, and to design the reversed flap for clinical reference. Methods Anatomic observation was performed on 30 adults’ lower extremity specimens perfused via pressure with red latex through femoral arteries. Results Superficial peroneal nerves, sural nerves and saphenous nerves were all nourished by their accompanying arteries which, anastomosed with the cutaneous perforating branches of other arteries, also nourish the corresponding skin areas. Conclusion The blood supply of the reversed island flap accompanying vessels of the cutaneous nerves in the lower leg is reliable and so it is possible to design long flaps along the cutaneous nervous axis.

Objective:To develop an experimental three-dimensional model by ECV304 cells(human umbilical vein endothelial cell line) for investigating the mechanisms of angiogenesis in vitro.Methods:ECV304 cells were seeded onto three-dimensional collagen gels made of rat-tail collagen.When the endothelial cells were cultured and grown to near confluence,treated with bFGF for 3 to 12 days,and then assessed with inverted phase contrast microscope.Results:The endothelial cells migrated into the gels,formed complex networks by cell cords at different levels through the bottom view,and sprouted capillary-like structures through the side view.Conclusion:ECV304 cells are capable of expressing some early events of angiogenesis in the three-dimensional collagen gels:proliferating,migrating and sprouting and so on.It should be useful for studying angiogenesis in vitro.