Objective To observe the expression of caspase-3 on the trehalose as cryoprotectant for preserving aortic valve homograft in liquid nitrogen.Methods The aortic valve homograft was divided into 5groups,namely:0.1 mol/L DMSO(control group),0.1 mol/L trehalose(experimental group 1),0.1 mol/L trehalose+0.1 mol/L DMSO(experimental group 2),0.2 mol/L trehalose+0.1 mol/L DMSO(experimental group 3),0.3 mol/L trehalose+0.1 mol/L DMSO(experimental group4).At the time of 12 months,15 months and 18 months when preserved in liquid nitrogen,relative expression of caspase-3 of the aortic valve homograft was measured by RT-PCR and Western Blot.Fresh group was a negative control group.Results At the same time(P<0.05),the expression of caspase-3 of fresh aortic tissue was slightest.The experimental group 2 was in accord with the experiment group 3,which was of a sort compare with the fresh group.The experimental group 4,which was worse than the experimental group 2 and 3,ranked above the experimental group 1.The worst was the control group.Conclusions The joint use of trehalose and DMSO could well inhibit the expression of caspase-3.Moreover.0.1mol/L trehalose+0.1 mol/L DMSO and 0.2 mol/L trehalose +0.1 mol/L DMSO could maximize the inhibition of the expression of caspase-3.

Objective To evaluate the effects of betulinic acid preconditioning on oxidative stress response during cerebral ischemia-reperfusion (I/R) in mice.Methods Seventy-two male Kunming mice,aged 3 months,weighing 25-35 g,were randomly divided into 3 groups (n =24 each) using a random number table:sham operation group (group S),I/R group,and betulinic acid preconditioning group (group BP).Cerebral I/R was induced by middle cerebral artery occlusion in mice anesthetized with 10％ chloral hydrate 40 ml/kg.In group BP,betulinic acid 50 mg/kg was administered by intragastric gavage everyday for 7 days before ischemia,while the equal volume of solvent dimethyl sulfoxide was given in S and I/R groups.At 22 h of reperfusion,neurological function was assessed and scored.The mice were then sacrificed and brains were removed for determination of infarct size,expression of NADPH oxidase (Nox1,Nox2 and Nox4) and p22phox mRNA,activity of ROS and apoptosis rate in the infarcted zone.Results Compared with S group,neurological score,cerebral infarct size,activity of ROS and apoptosis rate in the infarcted zone were significantly increased,and the expression of Nox1,Nox2,Nox4 and p22phox mRNA was up-regulated in group I/R.Compared with group I/R,neurological score,cerebral infarct size,activity of ROS and apoptosis rate in the infarcted zone were significantly decreased,and the expression of Nox1,Nox2,Nox4 and p22phox mRNA was down-regulated in group BP.Conclusion Betulinic acid preconditioning mitigates cerebral I/R injury through inhibiting oxidative stress response in mice.

OBJECTIVE To investigate the antitumor effect of cadmium (Ⅱ) complex of pyrazolone derivatives 1-phenyl-3-methyl-4-propionyl-5-pyrazolone salicyloyl hydrazide-cadmium (Ⅱ) (Cd-PMPP-SAL) on the murine melanoma B16 cells in vitro and in vivo and its mechanisms. METHODS B16 cells were incubated with Cd-PMPP-SAL at 1.0, 1.5, 3.0, 5.0 and 10.0 mg·L-1 for 24, 48 or 72 h. The prolifera? tion rate of B16 cel s was evaluated by MTT assay. B16 cel s were incubated with Cd-PMPP-SAL at 6.25, 12.50 and 25.00 mg·L-1 for 24 h, while cell morphology was observed by Hoechst33258 staining. Apop?tosis of B16 cells was detected by Annexin Ⅴ-FITC/PI staining. The activity of caspases in B16 cells was detected by caspase activity assay. C57BL/6J mice were inoculated subcutaneously with B16 cells to establish a tumor-bearing model. Five days later, Cd-PMPP-SAL at 6.25, 12.50 and 25.00 mg·kg-1 was injected into tumors of C57BL/6J mice once a day for 12 d. The body mass was recorded daily. One day after the last administration, all the mice were killed and the tumor was harvested. Tumor volume and mass were measured, and the tumor inhibitory rates were calculated. Pathological changes of the tumor, liver and lung were observed under a microscope. The expressions of vascular endothelial growth factor (VEGF) and fibroblast growth factor 2 (FGF2) in tumor tissues were detected by immuno?histochemistry. The apoptotic cells in transplanted tumor tissues were detected by TUNEL. RESULTS Cd-PMPP-SAL inhibited the proliferation of B16 cells. The IC50 was 4.946 mg · L-1, and 95% confidence interval was 4.24-5.65 mg · L-1. The apoptosis rates(12.8 ± 1.4)% and (18.4 ± 0.4)% of Cd-PMPP-SAL 12.50 and 25.00 mg · L-1 groups were significantly higher than those of control group (1.7 ± 0.1)% (P<0.01). The activity of caspase 3 and 9 of Cd-PMPP-SAL 25.00 mg · L-1 group was significantly higher than that of control group (P<0.01), but there was no significant difference in caspases 3/7. The relative tumor volumes of Cd-PMPP-SAL 6.25, 12.50 and 25.00 mg · kg-1 treated groups from the 8th day of treatment were significantly decreased compared with the model group (P<0.01). The result of paraffin sections showed that the transplanted tumor tissues in Cd-PMPP-SAL 12.50 and 25.00 mg · kg- 1 groups exhibited different degrees of necrosis, but there was no significant pathological damage to the liver or lung tissues of mice. Compared with model group, expressions of VEGF and FGF2 in Cd-PMPP-SAL 12.50 and 25.00 mg · kg-1 treated groups were significantly inhibited (P<0.05), and apoptotic cell rates were significantly higher (P<0.05). CONCLUSION Cd-PMPP-SAL can inhibit growth of B16 cells in vivo and in vitro, which may be associated with induction of tumor cell apoptosis and inhibition of tumor angiogenesis.

Objective:To compare the efficacy and safety between direct anterior approach (DAA) and posterior approach (PA) in total hip arthroplasty.Methods: This study evaluated postoperative results of 92 consecutive total hip arthroplasties performed by a single surgeon;44 from the DAA,and 48 from PA.The age,body mass index,operation time,blood loss,hospital stay,positioning of the artificial hip,postoperative Harris score and postoperative complications were recorded and analyzed.Results: Both the average age of the patients separately (58.0±11.9) years in DAA group and (61.0±10.4) years in PA group and the body mass index (25.1±3.7) in DAA group and (24.7±3.3) in PA group,showed no significant difference between the two groups.The DAA group had significantly reduced the hospital stay (3.8±1.7) days vs.(4.9±2.3) days for the PA group (P<0.05) and operation time was (76.0±17.4) min in DAA group,and (71.0±14.3) min in PA group (P>0.05).The amount of blood loss: in group DAA (238.0±55.3) mL,and in group PA (387.0±61.2) mL (P<0.05).There was no statistical difference in the positioning of the artificial hip: the cup anteversion in DAA group and PA group was 17.3°±5.3° vs.18.6°±5.1°,the cup inclination was 38.5°±5.7° vs.37.7°±5.2°.In DAA group,there was significantly less use of assistive devices [(24.6±7.8) d vs.(31.7±10.2) d,P<0.05],and the pain was significantly lower.Harris score at the end of 6 weeks of the follow-up: in DAA group 85.7±5.4,and in PA group 81.3±6.1 (P<0.05);at the end of the last follow-up: in DAA group 93.4±4.7,and in PA group 92.3±5.3 (P>0.05).Complications were encountered in the two groups.There were two intraoperative complications (4.4％),1 great trochanter fracture and 1 lateral cutaneous nerve injury in DAA group.No dislocation was observed in DAA group.One dislocations and 1 groin pain were recorded in PA group.No prosthesis loosening,deep vein thrombosis,sciatic nerve injury and other complications occurred in the two groups.Conclusion: Total hip arthroplasty using the anterior approach allows for superior recovery and better stability.

Objective To evaluate the effects of gestational weight gain(GWG) in different prepregnant body mass index (BMI) women on perinatal outcomes and to provide evidences for gestational weight management protocol.Methods Totally,2409 healthy singleton pregnant women accepted regular prenatal examinations in Nanjing Drum Tower Hospital from January 2009 to April 2010 were recruited in this study.They were divided into three groups according to pre-pregnant BMI,which were low BMI group (BMI＜18.5),normal BMI group (BMI 18.5-) and high BMI group (BMI≥24.0).According to GWG,the difference between pre-delivery maximal weight and prepregnant weight,the low and normal BMI women were divided into ＜10 kg,10 kg-and ≥15 kg GWG subgroups,and the high BMI women were divided into ＜5 kg,5 kg-,10 kg and ≥15 kg GWG subgroups.Data including gestational age,delivery modc,newborns' birth weight,Apgar score and incidences of gestational complications,such as hypertensive disorders complicating pregnancy (HDP),gestational diabetes mellitus (GDM),macrosomia,fetal growth restriction (FGR) and preterm birth,were recorded.Analysis of variance,Student-Newman Keuls,Chi-square test and Fisher exact test were applied for statistics.Results (1) Among the 2409 women,the percentages of low,normal and high BMI groups were 18.5％ (n=445),69.9％ (n=1685) and 11.6％ (n=279),respectively.The incidences of HDP,GDM,macrosomia and caesarean delivery in high BMI group were 12.9％ (n=36),17.9％ (n=50),13.6％ (n=38) and 52.3％ (n=146),respectively,higher than those in low BMI group [3.4％ (n=15),4.3 ％ (n=19),3.8％ (n=17) and 25.8％(n=115),x2 =23.8,37.1,23.5 and 50.2,P＜0.05] and those in normal BMI group [5.5％ (n=92),7.8％ (n=132),7.8％ (n=132)and 31.6％ (n=532),x2=21.8,29.0,10.1 and 3.4,P＜0.05].(2) In normal BMI group,the rates of FGR and preterm birth in GWG ＜10 kg subgroup were 3.5％ (4/115) and 8.7％ (10/115),higher than those in GWG 10 kg-subgroup [0.7％(4/548) and 3.3％(18/548),x2=6.0 and 6.9,P＜0.05] and GWG ≥15 kg subgroup [(0.8 ％ (8/1022) and 3.6％ (37/1022),x2=7.2 and 6.7,P＜0.05].The rates of macrosomia and cesarean delivery in GWG ≥15 kg subgroup were 10.7％ (109/1022) and 34.5％ (353/1022),higher than those in GWG＜10 kgsubgroup [3.5％ (4/115) and 32.2％ (37/115),x2=6.0 and 63.0,P＜0.05] and GWG 10 kg subgroup [3.5％ (19/548) and 25.9％ (142/548),x2=24.7 and 31.0,P＜0.05].(3) In high BMI group,the incidences of all pregnancy complications and perinatal outcomes did not show statistical significance among the four GWG subgroups (P＞0.05).Conclusions High prepregnant BMI is a high risk factor of pregnancy complications.It is suggested that normal BMI women should control GWG at 10-15 kg to lower the incidences of pregnancy complications.

BACKGROUND: The protein kinase C (PKC) family consists of 3 groups of PKCs, namely the conventional PKC (cPKC), atypical PKC and novel PKC.Accumulating studies conducted in recent years have suggested that PKCs may play important roles in the development of cerebral ischemic/hypoxic preconditioning ( I / HPC ).OBJECTIVE: To observe membrane translocation of hypoxia-activated cPKC isoforms(α, βⅠ, βⅡ and γ) at cellular levels in a cell hypoxia model.DESIGN: Randomized block design.SETTING: Department of Neurobiology, College of Basic Medical Sciences,Capital University of Medical Sciences.MATERIALS: The experiment was completed at the Neurobiological Cell Culture Laboratory of Capital University of Medical Sciences in May 2004.Human neuroblastoma cells with the properties of neurons were maintained and passaged in this laboratory.METHODS: The activation of cPKC isoforms under hypoxic condition and changes of cPKCα, βⅠ, βⅡ and γ membrane translocation(an indicator of PKCs activation) in SH-SY5Y neuroblastoma cells in response to hypoxia (1% 02, 5% CO2 and 94% N2) for 0 to 24 hours were observed using SDS-PAGE, Western blotting and immunocytochemistry.MAIN OUTCOME MEASURES: Effect of sustained hypoxia on cPKC membrane translocation in human neuroblastoma cells was observed with SDS-PAGE, cPKC Western blotting, and immunocytochemistry.RESULTS: cPKCα, βⅠ and βⅡ membrane translocation were increased significantly ( P ＜ 0.05 ) in a time-dependent manner in response to hypoxic exposure, and the increase of cPKCβⅠ was more evident( P ＜ 0. 001 ) after 4hours of hypoxic exposure, whereas no cPKCγ was detected in SH-SY5Y neuroblastoma cells either under normoxic or hypoxic condition. The results suggested that all cPKC isoforms, epically cPKCβⅠ, could be activated by sustained hypoxia, and the absence of cPKCγ in SH-SY5Y neuroblastoma cells may be relevant to the loss of specific biological features of the cultured cells.CONCLUSION: Sustained hypoxia activates the isoforms of cPKCα, βⅠ and βⅡ in human neuroblastoma cells and induces their membrane translocation.cPKCγ isoform may not exist in human neuroblastoma cells, or the cells has lost certain biological characteristics.

Literature about functional dyspepsia treated with acupuncture in recent 5 years is retrieved in China National Knowledge Infrastructure (CNKI), Wanfang database and PubMed. The research achievements are arranged and summed up to explore the mechanism of acupuncture for functional dyspepsia. It is found that acupuncture can regulate the secretion of braingut petide, and cause the coordination response of limbic system-brain. Also, it adjusts serum molecule metabolin and the gene expression of the transduction pathway of adjustment signal for rats. It is believed that functional dyspepsia treated with acupuncture is through multiple ways, and adjusting the function of braingut axis is one of the important mechanisms.
Acupuncture Points ; Acupuncture Therapy ; Animals ; Dyspepsia ; genetics ; metabolism ; therapy ; Humans ; Rats ; Signal Transduction

Objective To evaluate the diabetes self-management program based on Chinese local patients in Nanjing community. Methods From April 2014 to June 2014, diabetes patients were recruited through health records system screening in the community health service centers, letter invitation, poster announcements at communities, and telephone notification. A total of 53 self-management groups were established. Nanjing diabetes self-management program included six 1-1.5 hours sessions scheduled on consecutive weeks, based on the blueprint of Shanghai Chronic Disease Self-Management Program (CDSMP) developed at Stanford University. Baseline and three-month later interviews were conducted respectively. Results A total of 636 patients were recruited and agreed to enter CDSMP; 603 completed the 6-session activities, with the response rate being 94.8%. Compared to baseline, nine of the patients' the awareness rate of diabetes-related knowledge, six of self-management behaviors, the scores of quality of life in physical component summary [(47.51 ± 9.47) vs. (49.10 ± 8.27) points, t=6.170, P=0.000] and mental component summary [(47.09±11.95) vs. (49.13±10.74) points, t=5.157, P=0.000] were all higher after three months (all P values<0.05). Three months after implementation, the level of systolic blood pressure, diastolic blood pressure, fasting plasma glucose and total cholesterol decreased respectively by (1.42±0.52) mmHg (1 mmHg=0.133 kPa), (0.98 ± 0.34) mmHg, (0.66 ± 0.16) mmol/L, (0.15 ± 0.56) mmol/L,the differences were statistically significant (tpaired values were 3.935, 2.030, 4.889, 4.899, all P values<0.05). Conclusion The diabetes self-management program based on Chinese local patients for Nanjing may improve patients' awareness rate of diabetes-related knowledge, self-management behavior, the quality of life, and health status. CDSMP could be applied effectively in Nanjing.

We constructed prokaryotic recombinant expression vector of P61 gene from Nocardia brasiliensis,expressing P61 protein with biological activity in E.coli,and lay a foundation for further studies related to P61.P61 gene was synthesized and cloned into an expression vector pET-30a(+).The recombinant vector was transformed into Escherichia coli BL21 and induced with IPTG.The production was analyzed with Western blot and the catalase activity of P61 was tested with Catalase Assay Kit.The protein of P61was successfully expressed in E.coli with solubility and high catalase activity,and could be identified by anti-N.brasiliensis sera from mice.The prokaryotic expression plasmid of protein P61 was constructed successfully and can be expressed efficiently in E.coli BL21 cells with higher catalase.

Objective:To investigate the changes of c-Jun N-terminal kinase (JNK) and matrix metalloproteinase-9 (MMP-9) expressions after cerebral ischemia-reperfusion in rats and whether JNK inhibitor SP600125 can protect the cerebral ischemia-reperfusion injury by down-regulating the expression of MMP-9. Methods:Eighty-five adult male SD rats were divided into sham-operated group ( n=35), ischemia group ( n=35) and SP600125 group ( n=15). The middle cerebral artery occlusion models in rats of ischemia group and SP600125 group were established by thread embolism method, and reperfusion was performed one h after ischemia. Rats in the SP600125 group were injected with SP600125 solvent (being dissolved in 10% dimethyl sulfoxide with final concentration of 20 mmol/L) in the lateral ventricle 30 min before cerebral ischemia. At 48 h after reperfusion, neurological impairment scale scores, volume ratio of ischemic lesion, and moisture content of brain tissues in rats from each group were evaluated. The protein expression levels of phosphorylated (p)-JNK, JNK and MMP-9 in the ischemic cortex were detected by Western blotting in rats from the sham-operated group and ischemic group at 3, 6, 24 and 48 h after reperfusion, and in rats from SP600125 group at 48 h after reperfusion. Results:As compared with the sham-operated group, rats in the ischemia group had significantly decreased neurological impairment scale scores, and significantly increased infarction volume ratio and moisture content of brain tissues 48 h after reperfusion. At 24 and 48 h after reperfusion, the p-JNK/JNK values and MMP-9 protein level in the ischemia group were significantly increased as compared with those in the sham-operated group ( P<0.05). As compared with the ischemia group, at 48 h after reperfusion, rats in the SP600125 group had significantly increased neurological impairment scale scores, and significantly reduced volume ratio of ischemic lesion and moisture content of brain tissues, and significantly reduced p-JNK/JNK values and MMP-9 protein level in the ischemic cortex ( P<0.05). Conclusion:The expressions of JNK and MMP-9 are increased after cerebral ischemia-reperfusion, and the JNK inhibitor SP600125 may decrease MMP-9 expression to reduce the degrees of cerebral infarction after cerebral ischemia-reperfusion, thereby playing a role in brain protection.