1.The mechanism study of EGCG on improvement of cognitive function and alleviation of hippocampal insulin resistance in APP/PS 1 transgenetic mice
Chinese Journal of Biochemical Pharmaceutics 2014;(1):12-15
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.
2. Camellia nitidissima polysaccharides alleviate experimental intracerebral hemorrhage injury via regulating microglia M1/M2 polarization
Chinese Pharmacological Bulletin 2020;36(2):231-237
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.
3.Bionic biphasic ceramic bioactive bone for segmental bone defects
Ning DUAN ; Wentao ZHANG ; Huiguang CHENG ; Ning JIAO ; Shaofeng HAN
Chinese Journal of Tissue Engineering Research 2015;(25):3952-3956
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.
4.Effect of clemastine fumarate on lung ischemia-reperfusion injury in rabbits
Wei WANG ; Ning LIU ; Huirong HAN ; Ning LI ; Rui ZHANG
Chinese Journal of Anesthesiology 2016;36(10):1193-1196
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.
5.Research progress on the role of hedgehog signaling pathway in gastric cancer
Chinese Journal of Clinical Oncology 2014;(23):1540-1543
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.
6.Recent advances in PCR-based methods for the enrichment of minority alleles and mutations
Jinxia WU ; Ning HAN ; Hongwu BIAN
Chinese Journal of Laboratory Medicine 2014;37(12):958-963
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.
7.Screening and Identification of Mimotopes for Lipopolysaccharide Conservative Epitope from Random Phage Display Peptide Library
Weiyan WEN ; Qiangtao HAN ; Ning FU
Progress in Biochemistry and Biophysics 2001;28(2):222-226
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.
8.Construction of two retroviral vectors for interleukin-1 receptor antagonist and interleukin-10 and in vitro transfection of rabbit synoviocytes
Fei HAN ; Ning ZHANG ; Fulin TANG
Chinese Journal of Rheumatology 2001;0(02):-
Objectives To construct two retroviral vectors, one containing human interleukin-1 recep-tor antagonist (hIL-1Ra) gene and the other containing human interleukin-10 (hIL-10) gene and to transfect rabbit synoviocytes in vitro and detect the expression level of target genes. Methods RNA from human peripheral blood mononuclear cells were extracted and target genes were amplified by RT-PCR. The target genes were cloned into retroviral vector pLXSN, which was then transducted into GP2-293 cells to produce recombinant retrovirus. Rabbit synoviocytes were transfected and the expression of target genes was detected by RT-PCR, immunohistochemistry and western-blot. Results The retroviral vector containing hIL-1Ra gene or hIL-10 gene was constructed successfully. The hIL-1Ra gene and hIL-10 gene were transduced respectively into rabbit synoviocytes in vitro. The mRNA level of both genes reached peak in 5 days. In positive cell clones, the protein level of hIL-1Ra reached peak within 30 days and maintained at least 60 days; the protein level of hIL-10 maintained at least 40 days. Conclusion The hIL-1Ra gene and hIL-10 gene can be transduced successfully into rabbit synoviocytes by recombinant retrovirus.
9.Inhibitory effect of all-trans-retinoic acid on proliferation of human retinal pigment epithelial cells in vitro
Zhongqiu LI ; Yazhen WU ; Ning HAN
Journal of Jilin University(Medicine Edition) 2006;0(02):-
Objective To investigate the effects of all-trans-retinoic acid(ATRA) on proliferation of cultured human retinal pigment epithelium(RPE) cells and the probable mechanisms.Methods Cultured human RPE cells were treated with various concentrations(10-9,10-8,10-7,10-6 and 10-5 mol?L-1) of ATRA at different time points(6,12,24,48,72 and 96 h).Cell proliferation was evaluated by cell count and MTT colorimetric assay,and cell cycle analysis was performed by flow cytometry.Results The cell viability rates of ATRA treated group were decreased obviously,compared with control groups(P
10.Effect of sevoflurane pretreatment on expression of tight junction protein in the lung following ischemiareperfusion injury in rats
Jun CHAI ; Weimin CHEN ; Ning HAN
Chinese Journal of Anesthesiology 2010;30(5):612-614
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.