Objective To explore mechanism of epigallocatechin-3-gallate (EGCG) on improvement of cognitive function and alleviation of hippocampal insulin resistance in APP/PS 1 transgenetic mice. Method 12 months old female APP/PS 1 mice were randomly divided into 3 groups:model group(Tg), EGCG low dose group (Tg/EGCG-L), high dose group(Tg/EGCG-H). C 57 BL/6 J mice were utilized as control. learning and memory ability in 4 group mice were detected by morris water maze test(MWM). The hippocampal TNF-α/JNK signal and IRS-1 pSer 312 expression were detected by Western blot and immunohistochemical staining. Results Compared with NT mice, Tg mice showed a marked prolongation of the escape latency and swimming distance in the MWM test(P<0.05);Abnormal activation of TNF-α/JNK signaling and increased IRS-1 pSer 312 expression in the hippocampus of Tg mice(P<0.05). EGCG-treated Tg mice showed significantly improvement of all these abnormal changes(P<0.05). Conclusion EGCG treatment is able to alleviate hippocampal insulin resistance and improve cognitive function in the APP/PS 1 mice. which may be partly attributed to the reduction of TNF-α/JNK signaling activity in this AD mouse model.

Aim To explore the effect of camellia nitidissima polysaccharides (CNP) on acute intracerebral hemorrhage (ICH) and its mechanism related to regulation of microglia polarization. Method Adult male C57 BIV6 mice were randomly divided into sham-oprated control group, ICH group and CNP group. CNP was intragastrically administered immediately after intracerebral hemorrhagefor a consecutive three days. Neural functional outcomes were evaluated by neurological deficiency score (NDS) , open field test, and adhesive removal test. Blood-brain barrier destruction and pathological injury were detected by Evans blue staining and brain water content. Inflammatory factors were detected by ELISA. Microglia phenotypic status was evaluated and determined by quantitative real-time polymerase chain reaction ( qPCR) analyses, and immunofluorescence labeling. Results CNP significantly reduced neurological deficit scores and ameliorated cerebral edema and blood-brain barrier injury three days after ICH. Also, CNP treatment improved signifi-cantly motor function three days after ICH. In addition, CNP decreased proinflammatory mediators and inhibited the activation of microglia. Furthermore, treatment of CNP decreased microglia Ml markers and increased M2 markers. Conclusion CNP attenuates acute intracerebral hemorrhage thrdugh skewing microglia toward a more anti-inflammatory property.

Objective To investigate the effect of clemastine fumarate on lung ischemia-reperfusion (I/R) injury in rabbits.Methods Fifty New Zealand white rabbits of both sexes,weighing 2.0-3.0 kg,were divided into 3 groups using a random number table:sham operation group (Sham group,n =10),I/R group (n =20) and clemastine fumarate group (Cle group,n =20).The model of lung I/R was established by clamping the left hilum of lung and decreasing the tidal volume followed by restoration of perfusion and ventilation 1 h later in I/R and Cle groups.At 3 h of ventilation in group Sham and 2 and 4 h of reperfusion in I/R and Cle groups,blood samples were collected for determination of serum tumor necrosis factoralpha (TNF-α) and interleukin-8 (IL-8) concentrations by enzyme-linked immunosorbent assay.The left lung was lavaged,and the broncho-alveolar lavage fluid (BALF) was colleted for determination of white blood cell count.Lung specimens were obtained for microscopic examination of the ultrastructure of lung tissues and for determination of wet/dry weight ratio (W/D ratio),expression of IL-1β and IL-6 mRNA (by real-time polymerase chain reaction) and cell apoptosis (by TUNEL).The apoptosis rate was calculated.Results Compared with Sham group,the W/D ratio,white blood cell count in BALF,serum concentrations of TNF-α and IL-8 and apoptosis rate were significantly increased,and the expression of IL-1β and IL-6 mRNA was up-regulated in I/R and Cle groups (P＜0.05 or 0.01).Compared with I/R group,the W/D ratio,white blood cell count in BALF,serum concentrations of TNF-α and IL-8 and apoptosis rate were significantly decreased,and the expression of IL-1β and IL-6 mRNA was down-regulated in Cle group (P＜0.05 or 0.01).The pathological changes of lung tissues were significantly attenuated in Cle group than in I/R group.Conclusion Clemastine fumarate can attenuate lung I/R injury in rabbits.

BACKGROUND:The bioceramics has the ideal pore size, high porosity and the through-hole rate, can provide the ideal physiological activity space for the bone cel repair, and can obviously improve bone conduction. OBJECTIVE:To explore the bone conduction and bone induction in the repair of bone defects in the stage of bone defect of bionic biphasic ceramic bioactive bone. METHODS:A total of 20 New Zealand white rabbits were randomly divided into bioactive glass and biomimetic biphasic ceramic bioactive bone groups, and were used to construct the animal bone damage model. They were given the repair with bioactive glass and biomimetic biphasic ceramic bioactive bone. RESULTS AND CONCLUSION:At 4 weeks after model establishment, scanning electron microscopy demonstrated that dense periosteal tissue was observed in the biomimetic biphasic ceramic bioactive bone group. At 8 weeks, dense combination was found, and no obvious fissure existed. At 12 weeks, complete bone demarcation blurred, showing a natural transition. Moreover, the binding site was very dense. There were a large number of new bone tissues, bone trabecula was regular and connected to a piece. The bone material has been largely degraded. Bone defects were repaired completely. The bone density was close to normal bone. At 8 weeks, in the bioactive glass group, the binding site presented obvious fissure. At 12 weeks, the fissure had been connected, but the binding was not tight as compared with the bionic biphasic ceramic biologic active bone group. The bone defect got preliminary repair. A smal number of new bone formed trabecular bone, but could not connect or traverse. There was no recanalization of the marrow cavity. A few continuous bone cal us traversed the broken end. These data demonstrate that bionic biphasic ceramic bioactive bone has good bone conduction, bone induction and biocompatibility in the repair of segmental bone defects.

Objective To investigate the effect of sevoflurane pretreatment on the expression of tight junction protein Occludin and ZO-1 in the lung following ischemia-reperfusion (I/R) injury in rats. Methods Ninty-six adult male Wistar rats weighing 250-350 g were randomly divided into 4 groups ( n = 24 each): group Ⅰ sham operation (group S);group Ⅱ I/R; group Ⅲ sevoflurane (group Sevo) and group Ⅳ Sevo + I/R. The animals were anesthetized, tracheostomized and mechanically ventilated. Right femoral vein and left carotid artery were cannulated for BP monitoring, blood sampling and drug and fluid administration. Lung I/R was induced by clamping left pulmonary hilum for 45 min followed by 120 min reperfusion in group Ⅱ and Ⅳ. In group Ⅲ the animals inhaled 2.2 % sevoflurane for 30 min. In group Ⅳ the animals inhaled 2.2 % sevoflurane for 30 min before lung ischemia. Six animals were killed at the end of 45 min ischemia (T1), and 60 and 120 min of reperfusion (T2, T3 ). The lungs were immediately removed for determination of W/D lung weight ratio and the expression of Occludin and ZO-1 protein (by Western blot). Tissues of lung were obtained for observation of histopathology with light microscope. Lung permeability index (LPI) was calculated. Another 6 animals were killed at 120 min of reperfusion for lung lavage. Results I/R significantly increased W/D ratio and LPI and significantly reduced Occludin and ZO-1 protein expression in the lung tissue as compared with group C and Sev. Sevoflurane pretreatment significantly attenuated the I/R-induced changes. The pathological damage to the lung tissue was significantly less in group SP than in I/R. Conclusion Sevoflurane pretreatment may protect the lungs from I/R injury by up-regulating the expression of Occludin and ZO-1.

Objective To study the effects of extracorporeal shock wave therapy (ESWT) on avascular nec-rosis of the femoral head. Methods Thirty-two rabbits were used to make the avascular necrosis of the femoral headmodel by use of freezing method. Two rabbits were chosen to check the result of the model. Thirty rabbits were ran-domly divided into 2 groups : a treatment group and a control group. Each group was observed 2, 4 and 8 weeks aftertreatment. Morphological and pathological changes of the femoral head were observed. An immunohistochemicalmethod was used to examine the distribution of vessel endothelium growth factor (VEGF) and fibroblast growth factor(bFGF) immunoreactive-positive cells in the bone tissues. Results Morphologically, the femoral heads of treat-ment group were smoother and glossier than those of the control group. The empty lacunae ratio, the number of theosteoblast was significantly different between the treatment and the control group. The expression of VEGF and hFGFin the bone tissues of treatment group increased significantly when compared with control group. Conclusion ES-WT can promote healing of avascular necrosis of femoral head.

Hedgehog (Hh) signaling pathway is critical during embryonic development. Recent studies have shown that Hh sig-naling plays an important role in oncogenesis and development of many malignant tumors. In particular, activation of the Hh signaling pathway is implicated in the aggressiveness and progression of gastric cancer. This review provides an overview of the research prog-ress on the role of Hh pathway in gastric cancer. Investigation of this pathway may reveal a novel target for gastric cancer therapy.

Detection of low-level somatic DNA mutations and minority alleles within a wild-type sample is crucial in fields of medicine,such as cancer,prenatal diagnosis and infectious diseases.Increasing enrichment methods have been developed to study this challenging area.They are typically segregate by their ability to enrich for,and detect,either known or unknown mutations,including COLD-PCR,ice-COLD-PCR,TT-PCR,PNA-mediated PCR,WTB-PCR and digital PCR.In this review,we discuss theoretical principles and relative advantages and disadvantages of these techniques,and put an emphasis on their applications in fields of diseases.

To screen and identify the mimotopes for lipopolysaccharide(LPS) epitope, a rand om phage displayed dodecapeptide library was screened with a monoclonal antibody 2B4 spe ci fically against LPS conservative epitope. The positive clones were identified by phage ELISA and competitive inhibition assay by either S.typhi T 8-61 LPS or E.coli O111:B4 LPS. After three rounds of biopanning,the clones binding with 2B4 antibody were well enriched with positive rate of 80%. The bindings between 12 of positive phage clones and screening antibody were competitively inhibited by the two kinds of LPS,indicating that the positive clones have similar epitope with LPS. The positive peptide sequences were deduced from the corresponding DNA sequences. There were identical sequences among them. The seq uences were GPPQWFFSQPQL （5/12，41.7%），LPQYFWNTATTA （3/12，25%），FPQNHWNVP WAT（2/12，16.6%），HSQSFWNAPLAM and AHPWTHGYFPPL （1/12，8.3%） respectively . The results demonstrate that the peptides screened with 2B4 antibody are mimot opes for LPS conservative epitope.

Objective:To construct an immune inhibitor of new TNF-?,C-terminal sequence of new rhTNF-? was replaced with sequence of T-help cell of hen egg-white lysozyme(HEL).Methods:The rhTNF-? mutant was cloned 、expressed and purified.Results:The DNA sequencing analysis showed that the C-terminal sequence of new rhTNF-? mutant was correct.The mutant was inserted into pBV220 expression vector .After the recombinant bacteria was incubated at 42℃ for 4 h,a new band of the protein with relative molecular weight of 1.7 kD was shown on the gel.The band amounted to 30 % of total bacteria protein.Western blot showed that the mutant protein could associated with anti-TNF-? antibody.After the protein was purified by through a column of Q-Sepharose Fast Flow, the purity of the protein was above 90%.The biological activity of the protein was measured with L929 cells.The result showed that biological activity of the protein was totally lost.Conclusion:The experimental evidence demonstrated that the construction of new rhTNF-? mutant was successful.The mutant not only can be associated with anti-TNF-? antibody, but also lose the biological activity of original TNF-?. [