Objective To investigate the damage of hippocampal neurons induced by chronic cerebral ischemia in the rats,and to clarify its mechanism.Methods 90 healthy adult male SD rats were randomly divided into sham operation group (n=30 ) and experimental group (n=60 ). The chronic cerebral ischemia rat models were established by permanently ligating the common carotid arteries on both sides of the rats in experimental group.The rats in sham operation group were established by incising the cervical median,only the common carotid arteries on both sides were separated without ligating. The rats in sham operation group and experimental group were respectively sacrificed at the 7th,14th and 21st day after operation.At each time point 10 rats in sham operation group and 20 rats in experimental group were selected and sacrificed.Immunohistochemistry staining was used to observe the dynamic changes of the expression levels of choline acetyltransferase(ChAT)in hippocampus neurons, and TA-FE method was used to observe the dynamic changes of hippocampal microvascualr architecture. MiVnt image analytical system was used to quantitatively analyze the immunohistochemistry result,the microvessel density (MVD)and micorvessel area density (MVA)of horizontal part of hippocampus in the rats. Results Compared with sham operation group,the ChAT expression levels in hippocampus neurons of the rats in experimental group at different time points were significantly decreased(P<0.05);and with the prolongation of time,the ChAT expression levels were gradually decreased;the ChAT expression level in 14-day experimental group was significantly lower than that in 7-day experimental group (P<0.05 );the ChAT expression level in 21-day experimental group was significantly lower than those in 7-day and 14-day experimental groups(P<0.05).The MVD and MVA of hippocampus of the rats in experimental group at different time points were obviously decreased compared with sham operation group(P<0.05);the MVA was gradually decreased with the prolongation of time, and the MVA of hippocampus of the rats in 21-day and 14-day experimental groups were obviously decreased compared with 7-day experimental group(P<0.05);the MVD was gradually decreased with the prolongation of time,the MVD of hippocampus of the rats in 21-day and 14-day experimental groups was obviously decreased compared with 7-day experimental group(P<0.05);the MVD of hippocampus of the rats in 21-day experimental group was obviously decreased compared with 14-day experimental group(P<0.05).Conclusion Chronic cerebral ischemia can lead to the progressive decrease of the ChAT expression level,MVD and MVA of hippocampus of the rats to aggravate gradually the learning and memory dysfunction, which may be one of the reasons of vascular dementia.

Objective To investigate the mechanism of cytokines in brain pathologic changes caused by lipopolysaccharide(LPS).Methods A rat model of LPS was established by injection with a dose of 5mg/kg LPS into thigh vein.60 SD rats were randomly divided into three groups:control group,LPS group,naloxone+LPS group,each group included 20 rats and was subdivided into 4 subgroups(1h,2h,4h,6h respectively).In each subgroup,cerebral pathological changes and concentrations of TNF-α,IL-1β,IL-6 in brain homogenate were investigated.Results Brain homogenate concentration of TNF-α in subgroup LPS 2h was higher than control group[(9 30±0.57) ng/g vs(8.00±0 79)ng/g,P＜0.01],other subgroups have no significant difference from control group. Naloxone group had lower TNF-α level than LPS group.Brain homogenare concentration of IL-1β in subgroup LPS 1h increased apparently[(0.56±0.08)ng/g,P＜0.01],subgroup LPS 2h had no significant change;other subgroups have no significant difference from control group.Natoxone group had lower IL-1β level than LPS group(P＜0.01).but no difference from control group.Brain homogenate concentration of IL-6 in subgroup LPS 2h and 4h were respectively(459.68±79.15)ng/g,(816.07±111.31)ng/g,significantly higher than control group[(377.22±63.73)ng/g,(369.98±34.35)ng/g,respectively].Control group had no significant cerebral pathologic changes,while LPS group showed apparent changes:edema,interstitial substance and mening vessels expanding and hyperaemia,neurosome minification,Nissl's body lost,karyopycnosm,and nucleoli abolition.Naloxone group had lighter neurocyre injuries and brain edema,especially at 2h and 4h.Conclusions After LPS injection,the level of cytokines changed as time goes by,and this fits the brain pathologic changes.Naloxone has protecnve effect on brain tissue in LPS treated rats.Inflammatory cytokines play an important role in the mechanism of cerebral damage in LPS rats.

Objective To discuss application of evidence-based nursing in nasopharyngeal carcinoma patients undergoing radiotherapy. Methods Evidence-based nursing was used in 33 nasopharyngeal carcinoma patients undergoing radiotherapy. Results After evidence-based nursing, all the patients actively cooperated with the doctors and nurses, lived through the treatment process smoothly, and radiotherapy complications were reduced or avoided. Conclusions Evidence-based nursing is an idealized nursing method which could reduce or avoid the complications and alleviate the pain of patients and improve the quality of nursing.

Objective To investigate the inhibitory effect of immunosuppressive agent FRY720 on hepatocellular carcinoma Hepal-6. Methods Hepal-6 cells were cultured, and divided into 4 groups, namely control group and 0.1 μg/ml, 10 μg/ml, 100 μg/ml quality concentration groups. The cells were treated by the drugs for 24 to 48 hours respectively. The inhibitory rate of the cells was measured by MTT assay, and cell cycle and cell apoptotic rate were detected by flow cytometry (FCM). Results The ability of tumor cell growth were inhibited by FTY720 after 48 h. The maximal inhibition rate was 62.10 %, The apoptosis ratio was increased when FTY720 was 0.1-100 μg/ml, and it was 4.07 %, 8.16 %, 19.84 % respectively. FTY720 significantly prolonged cell G1 phase. Conclusion FTY720 could inhibit the growth of hepatocellular carcinoma, arrest the cell in G1 phase, and increase apoptosis.

Objective:To investigate the association between the apolipoprotein A5(APOA5) -1131T/C polymorphism and premature coronary heart disease in northern Chinese Han population. Methods: Using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and polyacrylamide gel electrophoresis (PAGE), we analyzed the genotype and allele distribution in 140 patients with premature coronary heart disease diagnosed by coronary angiography and 156 healthy controls. The levels of serum lipid profiles were also studied by biochemical methods. Results: The allele frequency of APOA5-1131T/C polymorphism in the premature coronary heart disease group was significantly higher (43.2% vs. 33.0%, P=0.011) than that in the control group. Compared with TT homozygotes, CC homozygotes exhibited a 2.809-fold (95% CI 1.331-5.927) increased risk of developing premature coronary heart disease. Logistic regression analysis found that this correlation was independent of sex, age, body mass index (BMI), smoking history as well as serum total cholesterol(TC), high density lipoprotein cholesterol (HDL-C) and low density lipoprotein cholesterol (LDL-C) levels; In premature coronary heart disease group, the triglyceride(TG) level in CC homozygotes was significantly higher than those in TC heterozygotes or TT homozygotes. Conclusion: The APOA5-1131T/C polymorphism has influence on serum TG level, and the APOA5-1131C allele is associated with the development of premature coronary heart disease in northern Chinese Han population.

T, p.R394W in exon 9.

Purpose To investigate the expression of IGF-Ⅱand IGFBP-6 in co1orecta1 cancer,and to exp1ore the c1inica1 significance in co1orecta1 cancer. Methods IGF-Ⅱand IGFBP-6 were detected by immunohistochemistry and RT-PCR in 50 cases of co1orecta1 cancer(experimenta1 group)and 50 cases of the adjacent mucosa(contro1 group). Results (1)In the experimenta1 group,the ex-pression 1eve1 of IGF-Ⅱprotein and mRNA was significant1y higher than the contro1 group. The expression 1eve1s of IGF-Ⅱprotein and mRNA in co1orecta1 cancer were significant1y 1ower than the contro1 group.(2)The expression 1eve1s of IGF-Ⅱand IGFBP-6 were sig-nificant1y different between different tumor infi1tration depth,1ymph node metastasis,invasion depth and Duke’s stages( P<0. 05), but no difference between genders,age and the degree of tumor differentiation( P>0. 05). Conclusions There is a obvious corre1a-tion between of IGF-Ⅱ and IGFBP-6 in c1inica1 patho1ogica1 parameters in co1orecta1 cancer. Combined detection of the two markers may be the bio1ogica1 indicators of occurrence and prognosis of co1orecta1 cancer,and provide a new scheme for diagnosis and treatment of co1orecta1 cancer.

Objective:To construct and express the fusion protein between ZZ affinity peptide and alkaline phosphatase and examine its biological activities.Methods:The alkaline phosphatase gene was cloned into pEZZ 18 vector containing ZZ peptide gene resulted in the pEZZ-AP recombinant vector.Then the vector was transformed and expressed in E.coli DH5α.And HisTrap affinity chromatography was employed to separate and purify the target protein.After analyzed by Western blot , ZZ-AP fusion protein was applied to immunocytochemistry as an alterative second antibody.Results:The result of SDS-PAGE showed that the fusion protein with a relative molecular weight was consistent with the theoretical values (62 kD).The fusion protein has rabbit IgG-binding ability and en-zymatic activity of alkaline phosphatase ,those were validated in Western blot;and it produced a good signal that was comparable in its staining pattern to that generated with goat anti-rabbit IgG-AP in immunocytochemistry.Conclusion: The ZZ-AP fusion protein was constructed successfully ,it has rabbit IgG-binding ability and enzymatic activity of alkaline phosphatase.We anticipate that the ZZ-AP fusion protein has a potential application in immunoassay field.

Objective To investigate the expression and significance of nuclear transcription factor-?B(NF-?B) in childhood acute lymphoblastic leukemia(ALL) and the effect of dexamethasone(DEX) on its expression,to provide the experimental base for corresponding clinical treatment of the ALL,in which NF-?B is taken as a target.Methods 1.The biotin-streptavidin method was used to detect NF-?B P65 protein on 20 childhood ALL patients and 20 healthy children.2.The effect of DEX at clinically relevant dosage on NF-?B P65 protein were also detected by the biotin-streptavidin method.Results 1.The positive expression rate of NF-?B P65 protein in childhood ALL patients was 85.50%,obviously higher than that in normal group(10.0%)(?~2=22.56 P

Objective To establish a human colon cancer multi-drug resistant cell line LS174T/5-fluorouracil(5-Fu) and to explore its biological characteristics. Methods A resistant human colon cancer line LS174T/5-Fu was established by stepwise increasing concentrations of fluorouracil selection from the parental cell line LS174T.Drug sensitivity was detected by MTT assay,and the changes of biological characteristics were determined by light microscopy,cell counting,flow cytometry and RT-PCR. Results LS174T/5-Fu cell line was developed after 6 months with stable resistance to 5-Fu and a resistance index of 40.24.The cells exhibited cross-resistance to cisplatin,etoposide,doxifluridine and hydeoxycamptothecin.LS174T/5-Fu cells grew more slowly than LS174T cells,which was longer than LS174T cell line.Cell cycle distribution of LS174T/5-Fu cells was changed compared with parental cells.The percentage of cells in G_(1) and S phase was increased,while in G_(2) phase was decreased.The level of MRP mRNA expression was increased according to the concentration of 5-Fu in resistant cell lines. Conclusion LS174T/5-Fu cells is a reliable multi-drug resistant cell subline of human colon cancer.The successful establishment of this cell subline has laid a solid foundation for further study of the multi-drug resistant mechanism of human colon cancer induced by 5-fluorouracil.