Objective To observe the protective effect of GSPB2 for delaying the human endothelial cells apoptosis with H2O2 induced human endothelial cells apoptosis as the model and to investigate its mechanism.Methods The human endothelial cells were pre-treated by 0.5 mmol H2O2,then different concentrations of GSPB2(5.0,10.0,20.0 μmol/L) was added for protecting cells.The cellular apoptosis was detected by TUNEL method;the apoptosis related molecules,such as Caspase-3,Bax,Bcl-2,and influencing apoptosis related molecules such as PI3K,p-Akt and Akt protein expression were detected by Western bl0t;Transwell chamber was used to detect cell migration situation.Results 0.5 mmol H2O2 induced obvious increase of human endothelial cells (P＜0.01).GSPB2 could significantly alleviate the H2O2 induced apoptosis of human endothelial cells(P＜0.05).10 μmol/mL GSPB2 could be close to return to H2O2 untreated control group level.Further found that Caspase-3,Bax protein expression were up-regulaed,Bcl-2 expression was down-regulated and the expression of PI3K,p-Akt and Akt wa sup-regulated.Conclusion The different concentrations of GSPB2 has a protective effect on vascular endothelial cells induced by H2O2,moreover which manifested by the concentration and time dependence.Its mechanism is related with the related molecules PI3K,p-Akt and Akt

Objective To study the relationship between cerebral vascular disease (CVD) and meteorological factors. Methods From 1998 to 2002, 6186 patients with CVD in Qingdao urban were studied. There were 1179 cases of intracerebral hemorrhage (CH), 4821cases of cerebral infarction (CI) and 186 cases of subarachnoid hemorrage (SAH). The relationship between the onset of CVD and meteorological factors including temperature, air pressure, relative humidity, wind velocity was analyzed by circular distribution analysis, Pearson correlation and multiple stepwise regression. Results High incidence of CH was found in winter and the mean angle was in January ((P

Objective To investigate the features and its risk factors for cognitive impairment in patients with lacunar infarct (LI) and white matter lesion (WML).Methods The inpatients with LI and WML aged 65 to 75 years old were enrolled.Their demographic and clinical data were collected.LI and WML were diagnosed by magnetic resonance imaging (MRI).Montreal Cognitive Assessment Scale (MoCA) was used to evaluate cognitive function.Self-Rating Depression Scale and Hamilton Anxiety Scale were used to exclude patients with depression and anxiety.The patients were divided into either a cognitive impairment group or a normal cognitive function group.The demographic and clinical data of both groups were compared.Multivariate logistic regression analysis was used to analyze and determine the independent risk factors for cognitive impairment.The characteristics of cognitive impairment of LI and WML were compared.Results A total of 130 patients with LI or WML were enrolled,92 of them had cognitive impairment,and 38 had normal cognitive function; 85 had LI,and 45 had WML; 53 were males and 77 were females.Univariate analysis showed that years of education in the cognitive impairment group (7.54 ±4.65 years vs.11.29 ±3.17 years; t =4.286,P=0.001) was significantly lower than that of the normal cognitive function group,while the constituent ratios of hypertension (54.6％ vs.16.2％ ;x2 =4.477,P =0.018),hyperlipidemia (53.1％ vs.16.2％ ;x2 =5.263; P =0.044),diabetes mellitus (46.9％ vs.10.8％;x2 =3.827,P=0.017),as well as LI (43.8％ vs.21.5％;x2 =3.928,P=0.015) and WML (26.9％ vs.7.7％ ;x2 =4.072,P =0.009) were significantly higher than those of the normal cognitive function group.Multivariate logistic regression analysis showed that years of education (odds ratio [OR],1.305,95％confidence interval [CI] 1.104-7.975; P =0.001),diabetes mellitus (OR 1.328,95％ CI 1.292-3.422;P =0.015),hypertension (OR 1.978,95％ CI 1.034-5.443; P =0.028,LI (OR 1.224,95％ CI 1.004-2.007; P =0.013),and WML (OR 1.489,95％ CI 1.202-3.778; P =0.010) were the independent risk factors for cognitive impairment.The total MoCA score (21.61 ± 5.33 vs.19.19 ± 7.07; t =1.841,P =0.038) and cube copy (0.43 ± 0.50 vs.0.31 ± 0.47; t =1.104,P =0.010),clock drawing test (2.53 ±0.89 vs.2.04 ± 1.22; t =2.229,P =0.008),letters identification (0.85 ±0.36 vs.0.62 ±0.50; t =2.585,P==0.000),and 100 minus 7 consecutively (2.62 ±0.79 vs.2.19 ± 1.17; t =2.113; P=0.001) of the WML group were significantly lower than those of the LI group.Conclusions The patients with LI and WML often had cognitive impairment,and the cognitive impairment in patients with WML was more serious.Years of education,hypertension and diabetes were the independent risk factors for cognitive impairment in patients with LI and WML.Visuospatial executive function and attention damage in patients with WML were severer than those of the patients with LI.

Objective To evaluate the clinical value of selective arterial infusion chemotherapy and embolization for rectal carcinoma.Methods Seldinger technique was used to insertsuperselectively catheter for 30 patients primary rectal carcinoma.Gelation embolization was done after infusionchemotherapy;All patients underwent operation in 7 day after chemoterapy and embolization.Results The clinical symptoms were obviously improved in all patients,25 in 30 patients were completely excised and the hemorrhage was obviously reduced.Conclusion The superselective arterial infusion chemotherapy and embolization may improve the clinical symptom and increase the resection rate in rectal carcinoma.

Objective To review and summarize the experience of surgical treatment of primary cardiac tumors in children from a heart center of China.Methods The pathological studies of 11 cases of primary cardiac tumor (except myxoma) were retrospectively analyzed.Results The mean age for all patients was (23 ± 16) months,and the average weight was 3.1 ～ 15.3 (13 ± 5.7) kg at the time of the established diagnosis.The most common reason of admission was the heart murmur,and the most common pathological type of cardiac tumors was rhabdomyoma (5 cases),followed by fibrous tumor (3 cases),angiofibroma (2 cases),and fibrosarcoma (1 case).All of the patients underwent tumor resection due to the corresponding symptoms.One of them,the operation findings showed the aortic root,superior vena and sinoatrial node were involved into the tumor body,and therefore,the tumor debark resection was performed and the child died 2 years later due to the remained tumor progression.One of other children (pericardial fibrosarcoma) died 1 year late because of tumor recurrence.The others were followed-up for duration of 2 ～9 years and all kept in good condition.Conclusions The surgical resection is mandatory for children suffering with symptomatic primary cardiac tumors and the short-term and long term surgical outcomes for the majority are acceptable.

Objective To investigate the effect of stellate ganglion block ~SGB) on formalin-induced nociceptive behavior of rabbits, the expression of brain-derived neurotrophic factor ~BDNF) of dorsal root ganglia ~DRG) and the concentration of serum interleukin-8 ~IL-8) in rabbits. Methods Twenty-four rabbits were used in this study. Catheters were inserted closely to right stellate ganglia of the animals, which were randomly divided into control group ~A), SGB group ~B) and pain model group ~C). Group B and C were administered with an injection of 0.5ml of 3% formalin into the right front paw subcutaneously, group A 0.5ml saline by the same way. 0.5ml of (0.25)% bupivacaine was administered via the catheter in group B at 10 minutes before injection, while group A and C were given 0.5ml of saline at the same time. Nociceptive behavior was observed for 60 minutes after injection, and assessed by weighted pain score. Serum IL-8 was measured radioimmunologically at 10 minutes ~T_0) before injection, 10 ~T_1), 60 ~T_2) and 120 minutes ~T_3) after injection. The expression of BDNF in the right DRG of cervical 7, 8 and thoracic 1 was investigated immunohistochemically at 120 minutes after injection. Results Formalin-induced nociceptive behavior in phase Ⅱ was relieved following SGB. The concentration of serum IL-8 in groups B and C at T_2 and T_3 was significantly increased than that at T_0 and T_1~P

BACKGROUND: Ketamine is a kind of frequently used general venous anesthesia drug in clinic, and the medication in vein or epidural cavum has analgesic effect. It is N-methyl-D-aspartic acid (NMDA) receptor noncompetitive antagonist, which can inhibit toxic effect of excitatory amino acids.OBJECTIVE: To observe effect of ketamine on apoptosis of dorsal horn astrocytes of spinal cord of rats induced by NMDA receptor over activation and explore its possible mechanism of action.DESIGN: Randomized controlled observation.SETTING: Department of Anesthesiology, Taihe Hospital Affiliated to Yunyang Medical College.MATERIALS: The experiment was conducted at Cell Biology Laboratory,Institute of Basic Medical Sciences, Yunyang Medical College between September 2003 and January 2005. Neonatal Wistar rats of two or three days were provided by Animal Experimental Center of Wuhan University. METHODS: Primary astrocytes in dorsal horn of T11-L6 spinal cord of Wistar rats were purified and cultured. Astrocytes were used in the experiment when its purity coefficient reached 98% assessed by gial fibrillary acidic protein. The cultured cells in 24-well plates were divided randomly into 6 groups (9 portions in each group): ①50 μL Hanks liquor was added into the control group. ②Amount of 100μmol/L was added into the NMDA group. ③Amount of 1 mmol/L was added into the ketamine group. ④100μmol/L NMDA + 0.1 mmol/L ketamine group. ⑤100 μmol/L NMDA + 0.5 mmol/L ketamine group. ⑥100μmol/L NMDA + 1 mmol/L ke tamine group. 1 mmol/L ketamine was clinical antalgic dosage. Activity of superoxide dismutase (SOD) and content of malondialdehyde (MDA) were examined after 24-hour culture. Content of Bcl-2 protein and change of morphology were observed with immunocytochemistry. Apoptosis of astrocytes was measured with flow cytometry. MAIN OUTCOME MEASURES: ① Counterstain cell staining and changes of morphology of Bcl-2 protein with immunohistochemical method and hematoxylin-esoin staining (HE). ②Apoptosis of astrocytes was detected with flow cytometry. ③Content of MDA and activity of SOD.RESULTS: ①Mean absorbance (A) of Bcl-2 as expression of Bcl-2 protein measured semiquantitatively: It was lower in the 100μmoL/L NMDA group than the control group, which had significant difference [0.054±0.021,0.108±0.039, respectively, P＜0.01]. It was higher in the 100 μmol/L NMDA + 1 mmol/L ketamine group than the 100 μmol/L NMDA group,which had significant difference [0.148±0.045, 0.054±0.021, respectively,P ＜ 0.01]. ②Apoptosis of astrocytes detected with flow cytometry: It was higher in the 100μmol/L NMDA group than the control group, which had significant difference [(25.26±6.13)%, (5.66±2.24)%, respectively, P＜0.01].It was lower in the 100μmol/L NMDA + 1 mmol/L ketamine group than in the 100μmol/L NMDA group, which had significant difference[(24.41±4.82)%, (25.26±6.13)%, respectively, P＜0.01]. ③Content of MDA and activity of SOD: 100 μmol/L NMDA made the content of MDA in astrocytes obviously increase , while the activity of SOD markedly decrease. 1 mmol/L ketamine remarkably decreased the content of MDA, distinctly increased the activity of SOD. This effectiveness had evidently dosage-effect relationship in clinical antalgic dosage, which had obviously difference as compared with that of the NMDA group (P ＜ 0.01 ). The differences between the 1 mmol/L ketamine group and the control group as well as between the 100 μmol/L NMDA + 0.1 mmol/L ketamine group and the NMDA group had insignificant difference.CONCLUSION: NMDA receptor over activation can induce apoptosis of a great number of astrocytes in spinal dorsal horn of rats. Suitable ketamine dramatically inhibits apoptosis, and its mechanism can enhance the expressionof Bcl-2 protein of astrocytes, at the same time inhibit the production of free radical and reinforce the activity of SOD.

AIM:To explose the possible existing pathway of intracellular signaling transduction in hypertensive induced by insulin in rat vascular smooth muscle cells proliferation which involved mitogen-activated protein kinase. METHODS:Male spontaneously hypertensive rat (SHR) aorta and WKY(6 weeks old) were isolated and then cultured to make the purified vascular smooth muscle cells.6-8th generation of VSMC were interfered with insulin in vitro . MAPK activity was determined by myelin basic protein method and its volume was measured with Western Blot. And [ 3 H]-TdR was used to measure DNA synthesis in VSMC proliferation. RESULTS: After the interfered with insulin the DNA synthesis was increased obviously in SHR group. MAPK activity and its contains in SHR were increased more than the control group. Protein kinase C inhibitor decreased MAPK activity induced by insulin. CONCLUSION:Proliferation of SHR VSMC in vitro was correlated with increased activity of MAPK. Insulin can affect MAPK induced activity. So an insulin-PKC-MAPK axis may exist in hypertensive VSMC.

Objective To explore the role of tumor inhibition of enhancer binding protein C/EBPα in the leukemic mice. Methods BALB/c nude mice were randomly divided into three groups. Three kinds of cells including pEGFP-C/EBPα-K562 cells, pEGFP-K562 cells and K562 cells as the control were injected into mice separately through the subcutaneous and tail vein, and subcutaneous tumors and leukemic models were formed. The changes of tumors were observed and the apoptosis of cells was detected by TUNEL; The capacity of proliferation of leukemia cells was observed in the bone marrow and the peripheral blood by Wright-Giemsa staining. The expression of genes of related to proliferation was detected by RT-PCR. Results The quality and the max diameter of tumors in the pEGFP-C/EBPα-K562 group were smaller than that of pEGFP-K562 group and K562 control group [(2.4±0.1) g vs (5.1±0.3) g and (5.7±0.4) g, both P ＜0.05; (11+2)mm vs (19+3) mm and (23+3) mm, both P ＜0.05]. More apoptosis cells were found in the pEGFP-C/EBPα-K562 group leukemic cells were found in the peripheral blood of leukemic models, and the proliferation of leukemic cells in the pEGFP-C/EBPo-K562 group were lower than that of other groups, accompany by the conspicuous cell differentiation. p53 was significantly elevated by RT-PCR, while down-regulated of c-myc.Conclusion Enhancer binding protein C/EBPα promote the apoptosis of cells and inhibit the proliferation of leukemia cells in leukemia mice, and further induce the cell differentiation. The inhibition of enhancer binding protein C/EBPα in the leukemia may have effect through the regulation of related genes.

Keeping an enzyme in its native form with high catalytic activity is of great significance. In the present study, thermal stabilizers of horseradish peroxidase (HRP) were screened. The results indicated that thermal stability of HRP was enhanced by magnesium sulphate and gelatin. A synergic effect of magnesium sulphate and gelatin was observed. In the presence of the stabilizer, the enzymatic activity of HRP remained 89% after kept for 80 h at 50 degrees C and 57% for 90 days at room temperature. Thermal alterations of HRP structure in the absence and presence of the stabilizers were explored by using UV absorption spectra at 402 nm (Soret band), intrinsic fluorescence and 8-anilinonaphthalene-1-sulfonic acid (ANS) fluorescence. The results suggested that magnesium sulphate and gelatin attenuated the extent of unfolding of HRP and therefore the native enzyme structure was stabilized.
Drug Synergism ; Enzyme Stability ; drug effects ; Gelatin ; pharmacology ; Horseradish Peroxidase ; metabolism ; Hot Temperature ; Magnesium Sulfate ; pharmacology