Objective To study the protective effects of propofol against ischemia-reperfusion injury in rat brains.Methods Modified Longa modle of focal cerebral ischemia-reperfusion injury was used. 200 healthy male SD rats, weighing 200-300g were anesthetized with intraperitoneal(I.P.) ketamine and propofol. When righting reflx was abolished, external carotid artery was exposed. A nylon thread with rounded end was inserted cranially until anterior cerebral artery was reached. After 3h ischemia nylon thread was withdrawn for reperfusion which lasted 3h. Bloos samples were obtained from orbit. Skull was opened and brain removed. In control group carotid artery was exposed but nylon thread was not inserted cranially. The animals were divided into four groups: (1)ischemia-reperfusion model group: normal saline 10 ml was administered I.P.,(2)operation control group: normal saline was given I.P.at the end of operation,(3)nimodipine group: nimodipine 1 mg?kg -1 was administered I.P. 10 min before ischemia,(4) propofol group: propofol 110 mg?kg -1 was given I.P. 10 min before ischemia. Brain infarction area, cerebral water content, serum lactate dehydrogenase(LDH) and creatine kinase(CK) levels,brain SOD activity and MDA and Ca 2+ levels were measured. Ultrastracture of brain tissue was examined by electron microscopy.Results Propofol 110 mg?kg -1 reduced mortality after brain ischemia/reperfusion injury. Infarction area of brain was significantly smaller in propofol and nimodipine groups than that in group 1. Propofol significantly inhibited the increases in serum LDH and CK levels induced by ischemia/reperfusion, increased SOD activity and decreased MDA content and Ca 2+ level in brain tissue. There was less brain tissue damage in propofol group.Conclusions Propofol 110 mg?kg -1 has protective effect against cerebral ischemia-reperfusion injury in rats.

A sequential extraction procedure has been proposed for the evaluation of the speciation of heavy metals including Cd,Cu,Pb and Zn in sediments from mariculture area,and the speciation of heavy metals was separated and defined as acid soluble fraction,reducible fraction,fraction bound organic matter,fraction bound sulfides and residual fraction. Matrix effects of high salinity on the determination of heavy metals in sediments were eliminated by matrix matching and internal standard methods when inductively couple plasma optical emission spectroscopy (ICP-OES) and mass spectroscopy (ICP-MS) were used,respectively. The results showed that the measured values of marine sediment reference materials were consistent with the standard values when the digestion solutions were determined after dilution. The extraction results of the prepositional extraction procedure and European Community Bureau of Reference Program (BCR) procedure were compared and the selectivity of extractants was investigated. The preliminary studies indicated that this sequential extraction procedure was applicable for evaluating the speciation of heavy metals in sediment with organic substances pollution and eutrophication,especially for fraction bound organic matter and fraction bound sulfides.

Objective To investigate the effect of theanine pretreatment on DNA repair function in neurons during brain ischemia-reperfusion (I/R) injury in rats.Methods One hundred and eight male Sprague-Dawley rats,weighing 290-310 g,aged 15 weeks,were randomly divided into 3 groups (n =36 each) using a random number table:sham operation group (S group),I/R group and theanine pretreatment group (T group).Global cerebral I/R was produced by 4-vessel occlusion method.Bilateral vertebral arteries were electrically cauterized,and bilateral common carotid arteries were clamped for 6 min.Theanine 1 g/kg was injected intravenously at 4 h before clamping bilateral common carotid arteries in T group,and the equal volume of normal saline was given in the other two groups.At 2,6,12,24,48 and 72 h of reperfusion,6 rats were selected in each group and sacrificed,the brains were removed,and the hippocampus was isolated for determination of the number of viable neurons in the hippocampal CA1 region (with a light microscope),apoptosis in neurons in the hippocampal CA1 region (by TUNEL),and expression of DNA repair protein X-ray repair cross-complementing group 1 (XRCC1) and Ku70 (by immunohistochemistry).The apoptotic index was calculated.Results Compared with S group,the number of viable neurons was significantly decreased,and the apoptotic index was significantly increased at 6,12,24,48 and 72 h of reperfusion,and the expression of XRCC1 and Ku70 was significantly down-regulated at 2,6,12,24,48 and 72 h of reperfusion in I/R group (P＜0.01).Compared with I/R group,the number of viable neurons was significantly increased at 12,24,48 and 72 h of reperfusion,the apoptotic index was significantly decreased at 6,12,24,48 and 72 h of reperfusion,and the expression of XRCC1 and Ku70 was significantly up-regulated at 2,6,12,24,48 and 72 h of reperfusion in T group (P ＜ 0.01).Conclusion The mechanism by which theanine pretreatment attenuates brain I/R injury is related to enhancement of DNA repair function and reduction of neuronal apoptosis in rats.

Objective To investigate if anodal transcranial direct current stimulation (atDCS) of the right cerebellum improves verbal working memory in amnesic persons with mild cognitive impairment (aMCI).Methods Thirty-nine aMCI were randomly divided into an observation and a control group using a random number table.The observation group was given atDCS at 1.2 mA for 20 minutes every day for 5 days,while the control group was provid ed with fake atDCS in the same way.Before and after the treatment,both groups were tested using forward and back ward digit spans,word reading,visually cued sensorimotor tests and finger tapping.Results After the treatment,the forward and backward digit spans of the observation group had improved significantly compared with before the treatment and with the control group's improvements.Significant improvement was observed in the average backward digit span of the control group,but not in their forward digit span after the treatment.No significant differences be tween the two groups were observed in the other measurements before or after the treatment.Conclusion Direct current stimulation of the cerebellum may improve the verbal working memory deficits of aMCI.Further research should be conducted to find the mechanism.

AIM: The effects of selenium dioxide (SeO_2) on proliferation, apoptosis, intracellular reactive oxygen species (ROS) and Ca~(2+) levels in three leukemia cell lines NB4, K562 and HL-60 were investigated. METHODS: Three leukemia cell lines were treated with 3-30 ?mol/L SeO_2. Flow cytometry was used to detect apoptosis rate, and analyze the changes of ROS and Ca~(2+) level within cells. RESULTS: SeO_2 at 10 and 30 ?mol/L inhibited proliferation in three leukemia cell lines. Treatment with 30 ?mol/L SeO_2 for 48 h induced 54.0%, 46.5%, 49.6% apoptosis in NB4, K562, and HL-60 cells, respectively, and also markedly decreased ROS and Ca~(2+) levels among three cell lines. The rate of ROS positive cells in NB4 and HL-60 decreased with the increase in SeO_2 concentrations. ROS was clearly reduced with 30 ?mol/L SeO_2 in K562. Ca~(2+) levels were tardily declined with 10, 30 ?mol/L SeO_2 in NB4 and HL-60 cells. Ca~(2+) levels were clearly reduced with 30 ?mol/L SeO_2 in K562. CONCLUSION: SeO_2 induces apoptosis in three leukemia cells. The declines of intracellular ROS and Ca~(2+) levels are involved in apoptosis induced by SeO_2.

This article summarizes the methods of making rat cerebral ischemia models and comments the advantages and disadvantages of various methods in order to provide references for the selection of animal models in the basis and appfication research of cerebral ischemia.

Objective To investigate the in vivo effects of allogeneic mesenchymal stem cells transplantation (MSCT) on OAZ signaling pathway in patients with systemic lupus erythematosus (SLE).Methods Isolated and expand human MSCs from bone marrow cells or umbilical cord of healthy donors were infused into SLE patients. Peripheral blood cells were collected from 10 pre-MSCT patients as well as 1 week and 4 week post-MSCT, and RNA was extracted and reverse transcripted to cDNA. mRNA expression levels of OAZ and Id1-3 were measured by using real-time PCR. Serum levels of IL-10, IL-12, IL-21, CCL2 and ANA were tested by ELISA. Relationships of the gene expression levels with levels of cytokines and ANA were analyzed. Results mRNA expression levels of OAZ, Id1 and Id3 gene in patients with SLE were significantly decreased at week 1(△C:12.4±1.1, 9.7±1.9, 9.7±1.9, 2.1±1.0) and at week 4 (△Ct:13.3±1.2, 10.4±1.5,10.8±1.2, 2.1±1.2) after MSCT when compared to those of the pre-MSCT (△Ct:11.0±0.9, 7.4±2.1, 7.8±2.1, 0.1±1.5 respectively, P all＜0.05). Levels of IL-10, IL-21 and ANA were significantly lower 4 week after MSCT than those before (P＜0.05); while level of CCL2 was higher than pre-MSCT (P＜0.05). Cytokines and ANA levels 1 week after MSCT were not differentially changed comparing to those of the pre-MSCT. Alteration of OAZ mRNA expression levels pre- and post-MSCT were negatively correlated with changes of ANA, IL-21levels and positively correlated with changes of IL-12/IL-10 and CCL2 levels. Conclusions The expression of genes involving in the OAZ signaling pathway is effectively reduced along with the alteration of several cytokines and ANA after allogeneic MSCT in SLE patients. OAZ signaling pathway may play an important role in MSCT treatment for SLE.

To develop a safe and effective new generation vaccine for IRKV-THChina12 prevention, we constructed a non-replicative recombinant human adenovirus carrying the IRKV-THChina12 G gene, named as rAd5-IRKV-G. The IRKV-THChina12 G protein expressed by the recombinant human adenovirus in 293AD cells was detected by western blot and indirect immunofluorescence test. To evaluate the immunogenicity of the recombinant, mice were immunized with rAd5-IRKV-G by intramuscular (i. m.) or intraperitoneal (i. p.) route and with non-exogenous gene expressing wild type adenovirus wt-rAd5 as a control. Results showed that the rAd5-IRKV-G could induce continuous and statistically significant (P ≤ 0.05) anti-IRKV neutralizing antibody (NA) production in immunized mice by i. m. or i. p. route. In particular, no significant difference (P > 0.05) of the NA titers between the two administration routes were observed, that provides an alternative choice for animal immunization method in the future application.
Adenoviruses, Human ; genetics ; physiology ; Animals ; Antibodies, Neutralizing ; immunology ; Antibodies, Viral ; immunology ; GTP-Binding Proteins ; genetics ; immunology ; Gene Expression ; Genetic Vectors ; genetics ; physiology ; Humans ; Immunization ; Lyssavirus ; enzymology ; genetics ; immunology ; Mice ; Rhabdoviridae Infections ; immunology ; virology ; Viral Proteins ; genetics ; immunology ; Virus Replication

Objective:To evaluate the effect of down-regulation of RACK1 expression on growth and radiosensitivity of oral squamous cell carcinoma cells.Methods:The shRNA vector for RACK1 gene was constructed and transfected into HSC-3 cells by lipofectamine. The stably-transfected cell line was obtained by constructing G418. The expression levels of RACK1 mRNA and protein were detected by RT-PCR and Western blot. The cell proliferation was detected by CCK8 assay. Cell apoptosis was examined by flow cytometry. The invasive and metastatic capabilities of cancer cells were assessed by cell invasion assay in vitro.The effect of X-ray irradiation combined with down-regulation of RACK1 expression upon cell proliferation was assessed by clone formation assay. The xenograft tumor nude mouse model was established to observe the inhibitory effect of down-regulating RACK1 gene expression combined with X-ray irradiation on oral squamous cell carcinoma. Results:RT-PCR revealed that the expression level of RACK1 mRNA of transfected HSC-3 cells was significantly down-regulated ( P<0.05). Western blot showed that the expression level of RACK1 protein was significantly down-regulated ( P<0.05). CCK8 assay demonstrated that down-regulation of RACK1 expression could remarkably inhibit the growth of HSC-3 cells ( P<0.05). RACK1 gene shRNA interference combined with X-ray irradiation significantly enhanced the apoptosis rate of HSC-3 cells ( P<0.05). The number of invasion cells in vitro in the RACK1 silencing group was evidently decreased ( P<0.05). Clone formation assay showed that the survival fraction in the shRACK1 group was significantly lower than that in the control group. The sensitization enhancement ratio was 1.37(ratio of D 0 value). Xenograft tumor experiment in nude mice showed that tumor growth was significantly inhibited in the shRACK1 group, the tumor volume was significantly decreased and the tumor mass was significantly lower than those in the control group (all P<0.05). Conclusion:Down-regulating RACK1 expression can enhance the radiosensitivity of oral squamous cell carcinoma cells, providing novel thinking to improve the radiosensitivity of oral squamous cell carcinoma.

Objectives To observe the effect of Luoyutong capsule on neurological function following focal cerebral ischemia-reperfusion in rats and to preliminarily study the protective mechanism of Luoyutong capsule for focal cerebral ischemia-reperfusion in rats. Methods A rat model of middle cerebral artery occlusion (MCAO)was induced by the modified Longa method. After 1. 5 h of ischemia,reperfusion started. Ten male SD rats were selected as sham operation group,and forty male SD rats were randomly divided into 4 groups:Model (MCAO),Luoyutong moderate-dose (LYTM),Luoyutong high-dose (LYTH),and citicoline sodium (CS)groups (n=10 in each group). At day 3 and 7 after modeling,the neurological function of the rats was evaluated by using 12 neurological score and forelimb placing test. Western blotting was used to detect the expression levels of brain derived neurotrophic factor (BDNF),basic fibroblast growth factor (b-FGF),and phosphor/protein kinase (p-AKT/AKT)on the ischemic side of the rats and in the ipsilateral brain tissue at day 3 after modeling,as well as the expression level of Caspase-12 at day 7 after modeling in the ipsilateral brain tissue,and a comparison was performed among the groups. Results (1 )Neurological score:At day 3 after modeling,there was no significant difference between the 12 neurological score and the forelimb placing test score (all P>0. 05);At day 7 after modeling, there were obvious improvement in the LYTM,LYTH,and CS groups compared with model group (all P<0.05). (2)The results of western blot showed that①compare with the sham operation group,the expression levels of BDNF and b-FGF were reduced obviously (all P<0.05);compare with the MCAO group,the expression levels of the LYTM,LYTH and CS groups could be up-regulated,particularly in the LYTH group (P<0. 01);② compare with the sham operation group,the expression level of p-AKT/AKT in MCAO group was decreased obviously (P<0. 05);compare with the MCAO group,the expression levels of p-AKT/AKT of the LYTM,LYTH,and CS groups were increased,particularly in the LYTH and CS groups (all P<0. 05);③ compared with the sham operation group,the expression of cleavage Caspase-12 was increased obviously in the MCAO group (P<0. 05). Compared with the MCAO group,the expression levels of proCaspase-12 and cleavage Caspase-12 had a decreasing trend in the LYTM and LYTH groups,but there were no significant differences (all P >0. 05);the expression levels of proCaspase-12 and cleavage Caspase-12 in the CS group were obviously lower than those of the MCAO group (P<0. 05). Conclusion Luoyutong capsule may play a protective effect for focal cerebral ischemia-reperfusion injury in rats by promoting neural survival and regeneration,and this protective effect may be associated with the inhibition of neuronal apoptosis.