ObjectiveTo observe the effect of nimodipine on hippocampal DNA-binding activity change of cAMP response element binding protein ( CREB )and CCAAT enhancer binding protein (C/EBP) in a rat model of vascular dementia(VD),and to explore the treatment mechanism of nimodipine.Methods66 healthy adult male SD rats were assigned to the following three groups of 22 each:VD model group,Sham-operated group,Nimodipine group.VD rat model was prepared by four-vessel occlusion.Physiological saline solution( 8 ml · kg-1 · d-1 )and Nimodipine (20 mg· kg-1 · d-1 )were administered by gavage respectively.The Morris maze was adopted to detect the changes of spatial learning and memorizing capacity,while HE straining was adopted to observe the changes of pathological characteristics in hippocampal CA1 area,and electrophoretic mobility shift assay(EMSA) were adopted to observe DNA-binding activity changes of CREB and C/EBP in hippocampus tissue.ResultsThe Morris maze showed:the learning and memory ability of nimodipine group rats ( escape latency period ( 26.63 ± 1.31 )s,the times of cross-platform(7.25 ±0.92) times) was higher than that of VD model group(escape latency period (41.25 ± 1.83 ) s,the times of cross-platform ( 5.33 ± 0.64 ) times ),with difference of statistical significance (P ＜0.05).HE results:in VD model group,neurons in CA1 were scaltered and boundaries were unclear,nuclei region was stained,coagulation necrosis appeared,obviously cells lost.The CA1 neurons of nimodipine group returned to be normal,nuclear membrane's profile and nudeolus were clear,regularly arranged; the number of hippocampal normal neurons in nimodipine group (43.19 ± 2.87 ) was more than that of VD model group( 16.33 ± 1.09 ),with difference of statistical significance(P＜0.05 ).EMSA:both CREB and C/EBP DNA-binding activity in rat hippocampus of nimodipine group ( ( 369.75 ± 13.22 ),( 428.25 ± 17.69 ) respectively ) were higher than those of VD model group ( ( 142.25 ± 27.86 ),(97.00 ± 5.88 ),respectively),with difference of statistical significance (P ＜0.01 )).ConclusionNimodipine can improve VD rats hippocampal neuronal injuries and their learning and memory impairment may be involved in the upregulating CREB and C/EBP DNA-binding activity.

ObjectiveTo investigate the effect of chronic stress on hippocampal neurogenesis.MethodsChronic unpredictable mild stress(CUMS) was used as animal model to provoke a decrease of neurogenesis in hippocampus.Simultaneously,hippocampal neurogenesis was monitored by assessing cell proliferation,survival,and differentiation.All rats were divided into control group,14 d CUMS group,28 d CUMS group.ResultsThe cell proliferation in 14d CUMS group(2254.17 ± 164.41 ) or 28d CUMS( 1900.33 ± 104.10) group were decreased contrast to the control (2919.50 ± 188.80) (P＜0.01).The cell survival in 28 d CUMS group ( 1845.33± 126.88 ) were decreased contrast to the control (2404.50 ± 148.77 ) (P ＜ 0.01 ).The cell differentiation had no difference between control group and CUMS group( NeuN/BrdU(71.63 ± 10.21 ) vs (69.11 ± 11.30),GFAP/BrdU ( 14.34 ± 2.03 ) vs ( 17.27 ± 2.93 ) ) (P ＞ 0.05 ).ConclusionThese results suggest that chronic stress decreases cell proliferation and survival,but does not affect the phenotype of newborn cells in the hippocampus.

AIM To observe the effect of Yishenjiangzhuo decoction on the NE,5-HT contents of hippocampus in cerebral ischemia reperfusion rats. METHODS High pressure lipuid chromatography-electrochemical process was used to measure the NE and 5-HT contents on the d 1, d 7 and d 15 after cerebral ischemia reperfusion by common carotid artery occlusion. RESULTS The NE contents of hippocampus were respectively 364.25?66.47, 349.76?59.38, (344.59?70.31) ?g?g -1 and the 5-HT contents were respectively 646.72?83.33,629.11?90.64,(596.68?99.47) ?g?g -1 on the d 1, d 7 and d 15 in the model group, which significantly decreased compared with the control group( P

Objective:To analyze the frequencies of HLA-A*0201 restricted CEA-specific CD8+T cells, HLA-A*0201/FLUmp tetramer and HLA-A*0201/CAP-1 tetramer were applied in patients with colorectal cancer.Methods: Lymphocytes from peripheral blood and lymph node,1×106 cells/ml,were incubated with 1μg HLA-A*0201/peptide tetramers and anti-CD8 for 1 h at 25 coseperately.The cells were then washed in PBS.Next,the cells were illuminated by detecting frequencies of FLUmp-specific CD8+T cells and CAP-1-specific CD8+T cells with flow cytometry.Results: HLA-A*0201/peptide were used to detect CAP-1 or FLUmp-specific CD8+T cells,which were analyzed either healthy individuals or patients with colorectal cancer.We did not find differences in average frequencies of FLUmp-specific CD8+T cells between 11 HLA-A*0201+patients with colorectal cancer and 14 HLA-A*0201+healthy individuals [ ( 0.671 ±0.421 )%, ( 0.564 ±0.408 )%].But the frequencies of CAP-1-specific CD8+T cells of HLA-A*0201+patients with colorectal cancer showed higher than HLA-A*0201+healthy individuals [ ( 2.409 ± 2.385 )%, ( 0.020 ± 0.021)%respectively],which was statistically significant(P=0.008).Conclusion:The frequencies of CAP-1-specific CD8+T cells in PBMC from peripheral blood and lymph node of HLA-A*0201+patients were increased,showed CEA-specific CTs has a vital role in colorectal cancer.

Objective To observe the effect of buyanghuanwu decoction on expression of immunoreactive protein and mRNA of NMDA receptor 2B subunit in rats hippocampal with vascular dementia to investigate the mechanism of buyanghuanwu decoction. Methods One hundred and forty-four rats were randomly divided into 4 groups: sham-operated group, VD model group,nimodipine group and buyanghuanwu decoction treatment group. The rats models of vascular dementia were built up by four vessels occlusion method. VD rats were treated with in-tragastrical buyanghuanwu decoction suspension (50 pharmacognostic g·kg-1·d-1) and nimodipine suspension (20 mg·kg-1·d-1) for 30 days. The learning and memory abilities were evaluated by Morris water maze tests. The change of NR2B protein in hippocampal of each group of rats were measured with immunohistochemistry and Western blot techniques and the expression of NR2B mRNA in hippocampus were observed by real-time fluorescence quantitative PCR techniques. Results Water maze tests,compared with sham-operated group((24. 18 ± 7.90)s,(7.99 ±1.32)/min) ,the escape latency(51. 25 ±18.28)s to explore the extension and the average spatial probe number ((5. 26 ±0. 74)/min) reduced in VD model group (P < 0. 05). Buyanghuanwu decoction ((25.91 ±9.56)s,(7. 52 ± 1. 27)/min) had significantly improved the above-mentioned rat model of learning and memory performance (P<0.05). There was no significant difference among sham-operated group,nimodipine group and buyanghuanwu decoction treatment group (P>0. 05). Similarly,as compared rats with sham-operated group(0.71 ±0.13), (5887 ±501), the expression of NR2B protein (0. 33 ± 0. 06) and its mRNA(593 ±53) were apparently decreased in VD rats (P< 0.05). The expression of NR2B protein(0.66 ±0. 11) and its mRNA (5692 ±482) in neuron of hippocampus were increased by buyanghuanwu decoction compared with the model group (P < 0. 05), and no difference was discovered between sham operation group and nimodipine group (P > 0.05).Conclusions Buyanghuanwu decoction improves the learning and memory abilities in VD rats, the therapeutic mechanism was concerned with lessening the injury of neurons on CA1 field in hippocampus and promoted the expression of NR2B protein and its mRNA.

Aim To investigate the effect of astragalus injection on the expression of JNK3(c-jun N terminal kinase)protein and JNK3 mRNA interrelated by apoptosis after hypoxia/hypoglycemia and reoxygenation in hippocampal neurons of rats.Methods The hippocampal neurons cultured for eight days were divided into four groups:normal control group,hypoxia/hypoglycemia and reoxygenation group,astragalus injection group and astragalus solution group.Hypoxia/hypoglycemia and reoxygenation group,astragalus injection group and astragalus solution group were treated with hypoglycemia and reoxygenation after being deprived of oxygen and glucose for 30 minutes.Methods of Western blot,ELISA and RT-PCR were used respectively to measure the expression of JNK3 mRNA after hypoxia/hypoglycemia and reoxygenation 0,0.5,2,6,24,72,120 h.Results Compared with normal control group,the mean optic density(MOD)of expression of JNK3 protein and activation of JNK3 protein in hippocampal neurons of rats every time points increased obviously in hypoxia/hypoglycemia and reoxygenation group except 120 h(P<0.05);compared with hypoxia/ hypoglycemia and reoxygenation group,MOD of expression of JNK3 mRNA and activation of JNK3 protein in hippocampal neurons of rats every time points decreased obviously except 120 h in astragalus injection group (P<0.05);compared with hypoxia/hypoglycemia and reoxygenation group,there was no difference in astragalus solution group.Compared with normal control group,MOD of expression of JNK3 mRNA in hippocampal neurons of rats every time points increased obviously in hypoxia/hypoglycemia and reoxygenation group(P<0.05);compared with hypoxia/ hypoglycemia and reoxygenation group,MOD of expression of JNK3 mRNA in hippocampal neurons of rats every time points decreased obviously in astragalus injection group except 120 h(P<0.05);compared with hypoxia/hypoglycemia and reoxygenation group,there was no difference in astragalus solution group.Conclusion Astragalus injection can inhibit the expression of JNK3 mRNA after hypoxia/hypoglycemia and reoxygenation,moreover,it can inhibit the expression of JNK3 protein and decrease the activation of JNK3 protein,accordingly it inhibits hippocampal neuronal apoptosis.

AIM:To observe the effect of buyanghuanwu decoction,a Chinese medicine,on the expression of AMPA receptor GluR1 subunit in mRNA and protein levels in rat hippocampus with vascular dementia (VD). METH-ODS:One hundred and forty -four rats were randomly divided into 4 groups:shamoperation group,VD model group,nimodipine group and buyanghuanwu decoction treatment group. The rat model of VD was built up by the method of 4 vessel occlusion. The VD rats were intragastrically treated with buyanghuanwu decoction suspension (pharmacognostic 50 g. kg -1.d -1) and nimodipine suspension (20 mg?kg -1.d-1) for 30 d. The learning and memory abilities were evaluated by Morris water maze testing. The change of GluR1 protein in hippocampal neurons in each group of rats was measured with immunohistochemistry and Western blotting techniques. The expression of GluR1 mRNA in hippocampus was determined by real -time fluorescence quantitative PCR. RESULTS:Compared to sham -operation group,the average escaping latency period (s) of Water maze tests in VD rats prolonged significantly and crossplatform time (numbers/min) shortened distinctly (P 0. 05). Compared to the rats in shamoperation group,the mRNA and protein levels of GluR1 were apparent-ly decreased in VD rats (P 0. 05). CONCLUSION:Buyanghuanwu decoction improves the learning and memory abilities in VD rats. The therapeutic mechanism is associated with lessening the neuron injury on CA1 field in hippocampus and restoring the mRNA and protein ex-pression of GluR1.

Objective To investigate the effects of astragalus injection on the morphology and expression of Apaf-1 in hippocampal neurons after cerebral ischemia reperfusion in rats. Methods The male SD rats were randomly divided into 3 groups, sham-operated group, cerebral ischemia-reperfusion group (reperfusion group) and astragalus injection interven-tion group (experiment group). The global cerebral ischemia-reperfusion rat model was established by Pulsinelli four-vessel occlusion method. The astragalus injection group was intraperitoneally injected with astragalus 6 mL/kg, 30 mins before sur-gery and repeated every 24 h. Rat brains were removed 24 h after reperfusion in each group. HE staining was used to observe the pathological changes of the hippocampal neurons under the light microscope. The ultrastructural changes of hippocam-pal neurons were observed by transmission electron microscopy. Immunohistochemistry and Western blot methods were used to measure the expression of apoptotic protease activating factor-1(Apaf-1) protein. Results Compared with sham-operat-ed group, nuclear and mitochondrial damage was found in reperfusion group, and the expression of Apaf-1 protein increased obviously in hippocampus(Immunohistochemistry result:0.024 ± 0.001 vs 0.109 ± 0.011;Western blot result:0.270 ± 0.018 vs 0.894±0.072, P<0.01). Compared with reperfusion group, the damage in nuclear and mitochondria was relieved obviously in experiment group, and the expression of Apaf-1 protein in hippocampus was significantly decreased (Immunohistochemistry result:0.048±0.005;Western blot result:0.392±0.046, P<0.01). Conclusion Astragalus injection can reduce pathological damage of hippocampal neurons after cerebral ischemia and reperfusion in rats, and the mechanism is related with inhibiting of Apaf-1 protein.

Objective: To study the epidemiological characteristics of tsutsugamushi disease, and to confirm the existence of the disease's epidemic foci in Taizhou. Methods: From 2013 to 2014, Dongxing town hospital and Xingqiao town hospital were selected as specimen collection sites in Jingjiang city. Blood samples (5 ml) were collected from 40 patients with acute tsutsugamushi disease. A total of 59 rodents were captured with cage night method in the survey sites at 5, 7, 9, 10, and 11 months in 2013, from which, the spleen, liver, and kidney specimens were selected. Chigger mites were captured by small blackboard method and from the ears of the captured rodents. A total of 226 small blackboards were laid, 27 mites were captured, and the samples were grounded into suspension. Nested-polymerase chain reaction and cell and tissue culture techniques were used to test the specimen from the probable patients, host animals and chigger mites. Results: Among the 40 acute tsutsugamushi disease blood samples, 29 were found to meet the test requirements, 17 were positive for orientia tsutsugamushi nucleic acid with 59% of the positive rate, and 1 stran orientia tsutsugamushi was isolated. 59 rats were captured and the density of mice was 5.5%. Among them, there were 26 Mus musculus (2.4%), 18 Rattus flavipectus (1.7%) and 15 Smelly shrew (density 1.4%). 1 Smelly shrew was tested positive for orientia tsutsugamushi nucleic acid, and the negative results were found in the other rodent specimens. 27 Chigge mites were collected by small blackboard method and the density of mites was 0.12 for each blackboard, among which 3 larvae and 24 nymphs were found. 33 Chigger mites were collected from the ears of 3 Smelly shrew, and the density of the mite was 11 per mouse. All the captured Chigger mites were identified as Leptotrombidium scutellare and 1 group of specimens of Chigger mites from the external environment were positive for orientia tsutsugamushi nucleic acid. Conclusion There was a high density of mice in the epidemic area from May to November and the species of the chigger mites were Chigger mites in Taizhou. The nucleic acid of the oriental tsutsugamushi was detected in the patients with acute scrub typhus, rodents and vectors. According to the above-mentioned results, it was considered that the scrub typhus epidemic area of Taizhou city has the natural foci of scrub typhus.