Preliminary analysis of species sensitivity distribution based on gene expression effect

Species sensitivity analysis is one of the major techniques applied to derive water quality criteria. Presently, the toxicity data used for development of water quality criteria are mainly in the biological individual level. With the increase of ecotoxicogenomics toxicity data, it is worth studying whether the gene expression effect data can be used to derive water quality criteria. Taking cadmium, copper and zinc as examples, we analyzed the toxic effects of the three heavy metals by constructing the species sensitivity distribution curves on the basis of extensive toxicity data. The results showed that the rank of species sensitivity for the acute, chronic and gene expression effect toxicity data of cadmium is "chronic>gene>acute". Although the gene expression effect data of copper and zinc are insufficient, the trend of data sensitivity of zinc is similar to cadmium. However, the trend of species sensitivity of copper is different from that of cadmium and zinc with higher sensitivity of gene expression data. It suggested that though the existing data of gene expression effects are not sufficient enough, they have the potential to be used in the development of chronic water quality criteria. For application in the derivation of water quality criteria, illogical test concentration design and insufficient target genes are two main weaknesses in the study of gene expression effects.

Effects of Pigment Indicators on the Color Expression of Leaves of the Colored-leaf Garden Plants

[Objective] In this study, the relationship between the pigments and the color expression of leaves of colored-leaf plants was discussed. [Method] The colors of leaf blades of 6 kinds of plants were analyzed with the Royal Horticultural Soci-ety Colour Chart. The chlorophyl content, carotenoids content and anthocyanin con-tent in leaf blades were determined. In addition, the color types of leaf blades, kinds of pigments, pigment contents and pigment distributions of 6 kinds of plants were compared. [Result] The chlorophyl contents ranked as Populus canadensis Moench （green leaves） 〉 Populus deltoids cv. Zhonghuahongye （purple green leaves） 〉 Populus euramericana cv. Quanhong （red leaves）; Acer palmatum Thunb. （green leaves） 〉 Acer palmatum cv. Atropurpureum （purple red leaves） 〉 Acer pal-matum Thunb. cv. Atropurpureum （red leaves）. The ranking of anthocyanin contents was just opposite. The chlorophyl content was negatively related to the anthocyanin content. The leaf color of plants is determined by various pigments. The more the chlorophyl is, the greener the leaf is. The more the anthocyanin is, the redder the leaf is. In the colored-leaf plants, the chlorophyl content represents about 80% of the content of pigments, the carotenoids content represents about 17%, and the an-thocyanin represents about 3%. There is a difference in the chlorophyl content be-tween colored-leaf plants and green plants. However, the relatively low chlorophyl content wil not hamper the normal life activities of colored-leaf plants. [Conclusion] We hoped to provide reference and basis for the production and landscaping of col-ored-leaf plants.

Interventional effect of hirudin on the expression of microtubule-associated protein 2 in peripheral tissue of hematom of model rats with acute intracerebral hemorrhage

BACKGROUND： It is suspected that dissociation, destruction or synthetic disorder of microtubule-associated protein 2 （MAP-2） may participate in secondary injury of intracerebral hemorrhage （ICH）, and the reason may be related to thrombin in high concentration after ICH; therefore, the mechanism should be studied further. OBJECTIVE： To explore the effect of hirudin on expression of MAP-2 in peripheral tissue of hematom after ICH and changes of water content in brain tissue and analyze pathogenesis of thrombin in secondary injury after ICH. DESIGN ： Completely randomized grouping design and controlled animal study SEn-ING ： Department of Neurology, the First Affiliated Hospital of Jilin University MATERIALS ： The experiment was carried out in the Neurological Laboratory of the First Affiliated Hospital of Jilin University from April 2003 to April 2004. A number of 80 healthy Wistar rats, of both genders, aged 3-4 months, weighing 250-350 g, were randomly divided into 8 groups： normal control group, 6-hour ICH group, 1-day ICH group, 2-day ICH group, 3-day ICH group, 7-day ICH group, 3-day hirudin group and 7-day hirudin group with 10 in each group. Five rats from each group were selected to measure their water content, and the others were undertaken immunohistochemical stain. Hirudin was produced by Sigma Company, USA, and MAP-2 rabbit-rat polyclonal antibody was provided by Fuzhou Maixin Biotechnology Company Limited. METHODS： ① Model establishing and grouping intervention： Rats in simple ICH group were collected their blood from tails and then inserted with 50 μL non-anticoagulant auto-arterial blood into the cauda of the putamen in right brain within 5 minutes. Rats in hirudin groups were inserted with 10 U hirudin （which was diluted with saline to 20 μL） into local hematom regions within 5 minutes, and the needle was pulled out after 10 minutes. Rats in normal control group were untouched. ② Water content in peripheral tissue of hematom： Based on the ratio between dry weight and wet weight, brain tissue at bleeding side and in right frontal lobe was selected to measure dry and wet weights so as to calculate the water content [（wet weight - dry weight） /wet weight] × 100%.③ Positive expression of MAP-2： Based on immunohistochemical stain, positive MAP-2 cells were regarded as neurons and they were buffy morphological. Positive rate of MAP-2 was calculated, i.e., percentage of positive cells in each sight to total cells in all sights. ④ Statistical analysis： Data among groups were compared with one-way analysis of variance, averages were compared with SNK-q test by each other, and relation between water content and MAP-2 was analyzed with linear regression technique. MAIN OUTCOME MEASURES： Changes of water content and MAP-2 expression in peripheral tissue of hematorn at various time points after ICH and intervention of hirudin. RESULTS： All 80 rats were involved in the final analysis. ①Water content： Water content was increased at day 1, reached peak at day 3 and decreased at day 7. It was （72.31±0.32）%, （77.42±0.53）%, （78.44±0.28）%, （74.10±0.13）%, （74.85±0.51）% and （70.07±0.36）%, respectively in 1-day, 2-day, 3-day and 7-day ICH groups and 3-day and 7-day hirudin groups, which was higher than that in normal control group （63.85±0.41, q=-4.684 3 to -7.262 0, P〈 0.05）; that in 2-day and 3-day ICH groups was higher than that in 7-day ICH group （q=-3.053 4, -3.727 0, P 〈 0.05）; and that in 3-day and 7-day ICH groups was higher than that in hirudin groups at the same time points （q=-2.965 6, -2.726 4, P 〈 0.05）. ②Positive expression of MAP-2： Positive expression of MAP-2 was decreased at 6 hours after ICH, reached the lowest value at day 3 and increased at day 7. Positive rate was （78.60±0.42）%, （60.56±0.74）%, （44.60±0.26）%, （25.45±0.85）%, （32.55±0.64）%, （37.69＋0.76）%, （41.75±0.68）%, respectively in 6-hour, 1-day, 2-day, 3-day and 7-day ICH groups and 3-day and 7-day hirudin groups, which was lower than that in normal control group [（96.50±0.33）%, q= -3.074 5 to -8.128 5, P 〈 0.05]. In addition, positive cells of MAP-2 disappeared plentifully at 3-7 days after ICH, stain of positive cells were light, and only stain of plasma was positive. That in 3-day and 7-day hirudin groups was higher than that in ICH groups at the same time points （q= -3.391 8, -2.967 9, P 〈 0.05）. Moreover, positive cells of MAP-2 was formed slightly but deeply stained. ③ Results of linear regression： Water content was negatively related to MAP-2 changes at 7 days after ICH （r= -0.894 9, P〈 0.01）, i.e., water content was increased with decrease of MAP-2 expression. CONCLUSION ： The deterioration of MAP-2 may be involved in the pathogenesis of thrombin within the first week after ICH, and the local administration of hirudin can protect neurons.

Phycocyanin for protecting brain ischemia-reperfusion injury and its effect on the expression of Caspase-3 mRNA

BACKGROUND ; Phycecyanin can anti-oxidize and clear free radial. Whether its protective effect on brain is related to Caspase-3, the promoter and operator of apoptosis, is highly concerned. OBJECTIVE： To observe phycocyanin for protecting nerve function and reducing the size of cerebral infarction of rats with brain ischemia-reperfusion and its effect on the expression of Cespese-3 mRNA. DESIGN ： A randomized controlled experiment. SETTING ： Institute of Cerebrovascular Disease, Affiliated Hospital of Medical College of Qingdao University MATERIALS： Totally 84 adult healthy female Wistar rats, weighing 210 to 250 g, of clean grade, were provided by the Animal Experimental Center of Shandong University. Phycocyanin （Institute of Oceanography of Chinese Academy of Sciences） was used. METHODS： This experiment was carried out in the Key Laboratory for Prevention and Treatment of Brain Diseases during May to December 2005. ① The rats were randomized into sham-operation group （n=4）, control group （n=-40） and phycocyanin-treated group （n=-40）. Middle cerebral artery occlusion/reperfusion （MACO/R） models were created on the rats of control and phycocyanin-treated groups with suture-occluded method by inserting a thread into left side extemal-internal carotid artery. In the sham-operatien group, inserting suture was omitted. After ischemia for 1 hour and reperfusion for 2 hours, suspension of phycocyanin was intragastdcaUy administrated into the rats of the phycocyanin-treated group at 100 mg/kg , and the same volume of normal saline was isochrenously administrated into the rats of control group as the same. ② Six rats were chosen respectively from the control group and phycocyanin-treated group, then neurologic impairment degrees of rats were evaluated according to Bederson＇s grading. ③ Six rats were chosen respectively from the control and phycocyanin-treated groups. The isolated brain tissue was stained with tdphenyltetrazolium chloride, and then the size of cerebral infarction was calculated with HPIAS-1000 image analytical system by calculating the ratio of cerebral infarction size at each layer and contralateral hemisphere size of the same layer. ④ Twenty--eight rats were chosen respectively from the control and phycocyanin-treated groups, Brain tissue was harvested at reperfusion for 6,12,24 hours and for 2,3,7 and 14 days after ischemia for 1 hour, respectively, 4 rats at each time point. Brain tissue of 4 rats of sham-opera- tion group was harvested at the 24^th hour after operation. Brain tissue sections were performed in situ hybridization detection of Cespase-3 mRNA. MAIN OUTCOME MEASURES： Comparison of neurologic impairment degree, cerebral infarction size and the expression of brain tissue Caspase-3 mRNA of rats between two groups RESULTS： Totally 84 rats entered the stage of result analysis. ① Bederson＇s scores at ischemia and reperfusion for 24 and 48 hours were significantly lower in the phycocyanin-treated group than in the control group（P 〈 0.05）. ② After brain ischemia and reperfusion, the infarction area was the largest in the 3^rc layer in both control and phycocyanin-treated group, which was（25.23±0,47）% and（23.09±120） %, respectively, and the size of infarction area in the 2^nd layer to the 5^th layer was significantly smaller in the phycocyanin-treated group than in the control group （P 〈 0.05）. ③Positive cell counts of brain tissue Caspase-3 mRNA： The number of positive cells of Caspase-3 mRNA of control group was increased from cerebral ischemia and reperfusion 6 hours, reached the peak at ischemia and reperfusion 24 hours, began to decrease 2 days later and positive cells of Caspese-3 mRNA were still expressed on the 14^th day after reperfusion. At ischemia and reperfusion 6,12 and 24 hours as well as 2,3,7 and 14 days, positive cell counts of Caspase-3 at peripheral ischemic area were significantly lower in the phycocyanin-treated greup[（70.67 ±3.65）, （85.06±4.79）, （119.54±5.37）,（74.26±2.19）, （62.06±3.34）, （23.11±1.89）, （10.75±2.63）/visual field] than in the control group [（94.38±8 28）, （108.81 ±16.11）, （140.88±14.47）, （98.13±11.31）, （81.03±9.31）, （31.22±8.86）, （16.06±5.96）Nisual field] （ P 〈 0.05）; and those at central ischemic area were also significantly lower in the phycocyanin-treated group [（33.86±4.01）, （39.51±3.46）, （50.96 ±2.53）, （43.07±4.09）, （36.25 ±3.72）, （9.03±3.87）, （4.91±5.59）/visual field ]than in the control group [（51.35±2.13）, （54.87±3.42）, （61.77±4.94）, （55.69±6.06）, （49.01 ±5.73） ,（12.84±3.37）, （7.32±2.39）/visual field]（P 〈 0.05）. CONCLUSION ： Phycocyanin can obviously improve the neurologic function, reduce the size of brain infarction and down-regulate the expression of Caspase-3 mRNA of rats with ischemia and reperfusion injury, thus protect brain.

THE EFFECT OF db-cAMP ON THE GENE EXPRESSION OF CALMODULIN AND CYTOSKELETON IN THE TRANSFORMED CELLS

We have demonstrated that the distribution of microtubules (MT), mlcrofilaments (MF) and fibronectin (FN) were diminished, while the gene expression of the calmodulin and c- fos enhanced in the transformed C3H10T1/2 cells. After treatment with 1 mM db-cAMP for 1 hour and 2 hours, there was an early and repldly reduced in gene expression of Calmodulin and c-fos respectively. After db-cAMP treatment for 4 -5 days, the number of capping cells of ConA binding decreased significantly and the cell surface microvllll decreased as well. The growth of treated cells was inhibited markedly. By using 4F1 cDNA probe, which is preferentially expressed In G1 phase, we have found that the db- cAMP treated cells were accumulated at G1 phase. Of particular interest is the fact that the distribution of microtubules, mlcrofilaments and fibronectln were recovered after treatment with 1 mM db-cAMP for 6 days. It is suggested that the Inhibition of proliferation, alteration, of phenotype and reco- very of cytoskeleton is transformed cells after treatment with db-cAMP are related to the Inhibition of gene expression of Calmodulin.

Genetic expression and effects of the rolled leaf gene Rl(t) in hybrid rice (Oryza sativa L.)

We constructed a nearisogenic line of rolled leaf gene Rl(t), which ex-pressed incompletely dondnance for the character of rolled leaf(RL), with ge-netic background of Zhenshan 97B. Using RL Zhenshan 97B and the originalZhenshan 97B as the female parents, and Minghui 63 and Yanhui 559 as themale parents, crosses of RL Shanyu 63 (RS63) and Shanyu 63(S63), RLShanyou 559 (RS559) and Shanyou 559 (S559) were made. Inheritance andeffects of Rl(t) in hybrid rice were studied at the flowering and at the 20 d afterflowering, respectively. Results were as follow:

Suppressive effect of dexamethasone on the neutrophil expression of CD18 in rats with radiation induced brain edema

BACKGROUND： Stereo-tactic radiation therapy （SRT） is widely used to treat intracranial diseases, but some patients suffered from radiation induced brain edema after SRT. Once radiation induced brain edema occurs, the treatment is quite difficult, and it always leads to a poor outcome. Dexamethasone has certain therapeutic effect on traumatic brain edema, but the biological mechanism is still unclear. OBJECTIVE ： To observe the effect of dexamethasone on the neutrophil expression of CD18.DESIGN ： A randomized control observation.SETTING： Changhai Hospital of the Second Military Medical University of Chinese PLA. MATERIALS ： The experiment was carried out in Changhai Hospital of the Second Military Medical University of Chinese PLA from January 1999 to December 1999. Twenty SD rats （male and female each in half） weighing （250±50） g were used. METHODS： Twenty SD rats were divided into four groups at random. ① Blank control group （n=5）： The rats were not treated without dexamethasone or irradiation;② Irradiation group （n=5）： The rats were given irradiation but no dexamethasone treatment; ③ Irradiation＋1 mg/kg dexamethasone group （n=5）; The rats were treated with irradiation and dexamethasone of 1 mg/kg; ④Irradiation＋5 mg/kg dexamethasone group （n=5）： The rats were treated with irradiation and dexamethasone of 5 mg/kg. The heads of the rats were irradiated with 10 MeV X-ray （30 Gy）, and brain tissue was removed after 2 weeks to observe the pathological changes. Blood samples were taken from the carotid artery, gradient centrifugation was used, and neutrophile layer was obtained, the level of neutrophile expression of CD18 mRNA and quantity of membrane proteins in blood were detected with Northern blot and flow cytometry respectively. MAIN OUTCOME MEASURES： ① Blood cell count; ② Pathological results; ③ level of neutrophile expression of CD18 mRNA and quantity of membrane proteins. RESULTS ： All the 20 SD rats were involved in the analysis of results without deletion. At 2 weeks after irradiation, obvious cell injury could be observed under light microscope. The level of neutrophile expression of CD18 mRNA and quantity of membrane proteins in blood were obviously increased, but the severity of cell injury was relieved in the irradiation＋1 and 5 mg/kg dexamethasone groups, and the CD18 expression was markedly suppressed （P 〈 0.05）, and the suppression was more obvious in the irradiation＋5 mg/kg dexamethasone group than in the irradiation＋1 mg/kg dexamethasone group （P 〈 0.01 ）. CONCLUSION： Dexamethasone can reduce the radiation induced brain edema by inhibiting the expression of CD18.

Effects of phycocyanin on apoptosis and expression of superoxide dismutase in cerebral ischemia reperfusion injury

BACKGROUND ： The application of exogenous antioxidant is always the focus in the prevention and treatment of cerebral ischemia. Phycocyanin has the effects against oxidation and inflammation, but its role in the pathophysiological process of cerebral ischemia reperfusion injury still needs further investigation. OBJECTIVE： To observe the effects of phycocyanin on the expression of superoxide dismutase （SOD） apoptosis and form of the nerve cells in rats after cerebral ischemia reperfusion injury. DESIGN： A randomized control animal experiment SETTING ： Institute of Cerebrovascular Disease, Medical School Hospital of Qingdao University MATERIALS： Fifty-two healthy adult male Wistar rats of clean degree, weighing 220-260 g, were used. Phycocyanin was provided by the Institute of Oceanology, Chinese Academy of Sciences. METHODS： The experiments were carried out in Shangdong Key Laboratory for Prevention and Treatment of Brain Diseases from May to December 2005. ① All the rats were divided into three groups according to the method of random number table： sham-operated group （n=4）, control group （n=24） and treatment group （n=24）. Models of middle cerebral artery occlusion/reperfusion （MCAO/R） were established by the introduction of thread through external and internal carotid arteries in the control group and treatment group. After 1-hour ischemia and 2-hour reperfusion, rats in the treatment group were administrated with gastric perfusion of phy- cocyanin suspension （0.1 mg/g）, and those in the control group were given saline of the same volume, and no treatment was given to the rats in the sham-operated group. ②The samples were removed and observed at ischemia for 1 hour and reperfusion for 6 and 12 hours and 1, 3, 7 and 14 days respectively in the control group and treatment group, 4 rats for each time point, and those were removed at 1 day postoperatively in the sham-operated group. Forms of the nerve cells were observed with toluidine blue staining. Apoptosis after cerebral ischemia reperfusion was detected with TUNEL technique. SOD expression was detected with immunohistochemical technique.③ The intergroup difference was compared with the ttest. MAIN OUTCOME MEASURES： The apoptosis of the nerve cells and SOD expression were mainly observed in each group. RESULTS： Finally, 52 rats were involved in the analysis of results. ① Number of apoptotic cells： In the sham-operated group, a few apoptotic cells could be observed in brain tissue. The apoptotic cells at each time point in the control group and treatment group were obviously more than those in the sham-operated group （P 〈 0.05）. In the treatment group, the numbers of apoptotic cells at 12 hours, 1 and 3 days after reperfusion were significantly fewer than those in the control group, and those at 6 hours, 7 and 14 days were similar to those in the control group. ② Number of SOD positive cells： In the sham-operated group, there was weak expression of SOD in brain tissue, and the positive cells were extremely few, the positive cells at each time point were significantly more in the control group and treatment group than in the sham-operated group （P 〈 0.05）. In the treatment group, the numbers of positive cells at 6 and 12 hours, 1 and 3 days after reperfusion were significantly fewer than those in the control group, and those at 7-14 days were similar to those in the control group. ③ Cellular forms： In the control group, the karyopyknosis occurred in the nerve cells, which were irregularly distributed, nucleolus disappeared, and some scattered cell fragments were observed. The forms of the nerve cells in the treatment group were generally normal. CONCLUSION ： Phycocyanin plays a neuroprotective role in cerebral ischemia reperfusion injury by activating the SOD expression and inhibiting apoptosis.

The effect of laminar shear stress on the expression of matrixmetalloproteinases-9 in rat MSCs

To investigate the effects of mechanical factors on matrix metalloproteinase9(MMP-9) expressions in rat bone marrow-derived mesenchymal stem cells(MSCs) and possible mechanism signal.Rat bone marrow MSCs were isolated and cultured,then,exposed to laminar shear stress

Effect of Ligustrazine on Expressions of Basic Fibroblast Growth Factor and Its Receptor in Bone Marrow of Mice with Acute Radiation Injury

Objective: To study the expressions of basic fibroblast growth factor (bFGF) and its receptor (bFGFR) in bone marrow of mice with acute radiation injury, and to evaluate the effect of Ligustrazine (Lt) on them. Methods: Fifty-six Kunming mice of clean grade were randomly divided into 3 groups, the normal group, the control group and the Lt group. Mice in the latter two groups were once homogeneously systemic irradiated with 6.0 Gy of 60 Co, with the absorption dose rate of 0. 56 Gy/min, then treated with saline (0.2 ml/ mice) or Lt (2 mg/mice) respectively, twice a day through gastrogavage for successive 13 days. Mice were sacrificed in batch on the 3rd, 7th and 14th day by cervical dislocation to collect the bilateral femoral bone marrow for preparing bone marrow mono-nuclear cell (BMMNC) suspension. The bFGFR expression on surface of BMMNC was determined by flow cytometry; and the bFGF expres-sion level in one side of femoral bone marrow tissue was detected by immunohistochemistry with SABC-AP assay. Results: The bFGF expression in bone marrow of mice on the 3rd, 7th and 14th day after acute radiation injury all were significantly lower than that of the normal mice (P<0.05 or P<0.01). The expressions of bFGF and bFGFR in the Lt group detected were significantly higher than that in the control group detected at the corresponding time points (P<0.05 or P < 0.01). Conclusion:By way of enhancing bFGF expression in bone marrow and bFGFR expression on surface of BMMNC to accelerate the repairing of hemopoietic micro-environment in bone marrow might be one of the mechanisms of Lt in promoting hemopoietic function reconstitution after acute radiation injury.Original article on CJITWM (Chin) 2004;24(5):439

Effect of ketamine on aquaporin-4 expression and neuronal apoptosis in brain tissues following brain injury in rats

BACKGROUND： Aquaporin-4 （AQP-4） is closely related to the formation of brain edema. Neuronal apoptosis plays an important part in the conversion of swelled neuron following traumatic brain injury. At present, the studies on the protective effect of ketamine on brain have involved in its effect on aquaporin-4 expression and neuronal apoptosis in the brain tissues following brain injury in rats. OBJECTIVE： To observe the effect of ketamine on AQP-4 expression and neuronal apoptosis in the brain tissue following rat brain injury, and analyze the time-dependence of ketamine in the treatment of brain injury.DESIGN： Randomized grouping design, controlled animal tria SETTING ： Department of Anesthesiology, the Medical School Hospital of Qingdao University MATERIALS： Totally 150 rats of clean grade, aged 3 months, were involved and randomized into control group and ketamine-treated group, with 75 rats in each. Each group was divided into 5 subgroups separately at 6,12, 24, 48 and 72 hours after injury, with 15 rats at each time point. Main instruments and reagents： homemade beat machine, ketamine hydrochloride （Hengrui Pharmaceutical Factory, Jiangsu）, rabbit anti-rat AQP-4 polyclonal antibody, SABC immunohistochemical reagent kit and TUNEL reagent kit （Boster Co.,Ltd., Wuhan）. METHODS： This trial was carried out in the Institute of Cerebrovascular Disease, Medical College of Qingdao University during March 2005 to February 2006. A weight-dropping rat model of brain injury was created with Feeney method. The rats in the ketamine-treated group were intraperitoneally administered with 50 g/L ketamine （120 mg/kg） one hour after injury, but ketamine was replaced by normal saline in the control group. In each subgroup, the water content of cerebral hemisphere was measured in 5 rats chosen randomly. The left 10 rats in each subgroup were transcardiacally perfused with ketamine, then the brain tissue was made into paraffin sections and stained by haematoxylin and eosin. Neuronal morphology was observed. AQP-4 expression and neuronal apoptosis were measured with immunohistochemical method and TUNEL method respectively. MAIN OUTCOME MEASURES： Water content in brain tissue, neuronal morphology, the number of AQP-4 positive neurons and TUNEL positive neurons in rats of two groups at each time point after injury. RESULTS： Totally 150 rats entered the stage of result analysis. （1） Water content of brain tissue： The water content of brain tissue at each time point after injury in the ketamine-treated group was lower than that in the control group. There were very significant differences in water content at 12 and 24 hours after injury respectively between ketamine-treated group and control group [（77.34±2.35）% vs. （82.31 ±1.48）%; （78.01 ±2.21 ）% vs. （83.86±2.37）%, t=-4.001 6,4.036 7, both P 〈 0.01]. （2） Neuronal morphology： Pathological changes in traumatic region and peripheral region of injury in the ketamine-treated group were significantly lessened, and necrotic and apoptotic cells in the ketamine-treated group were also significantly reduced as compared with control group. （3） AQP-4 expression： AQP-4 positive neurons at each time point in the ketamine-treated group were significantly less than those in the control group. There were very significant differences in AQP-4 expression at 12 and 24 hours after injury between ketamine-treated group and control group [（34.17±4.74） /visual field vs. （43.42±5.65） /visual field;（40.83±3.17） /visual field vs. （58.88±6.23） /visual field,t=3.966 3,8.165 7, both P〈 0.01]. （4） Neuronal apoptosis： TUNEL positive neurons at each time point in the ketamine-treated group were less than those in the control group. There were very significant differences in the neuronal apoptosis at 12 and 24 hours after injury between ketamine-treated group and control group [（26.25±3.04） /visual field vs. （32.75±4.39） /visual field; （29.33± 4.02） /visual field vs. （39.83±5.61） /visual field,t=-3.849 3,5.169 2,both P 〈 0.01]. CONCLUSION： Ketamine can reduce brain edema, AQP-4 expression and neuronal apoptosis following brain injury in rats, and has obvious therapeutic effect on brain injury, especially at 12 and 24 hours after injury.

Sexual differences of the effects of prenatal stress on the expression of extracellular signal-regulated kinaseas in the hippocampus of offspring rats

BACKGROUND： Prenatal stress has been shown to inhibit cell proliferation in the dentate gyrus and hippocampus, reduce hippocampal volume, and cause neuronal loss and oxidative damage in the hippocampus of offspring rats, but the sexual difference of the effects on offsprings is seldom referred to. OBJECTIVE： To observe the effect of prenatal stress to adult pregnant rats on expression of extracellular signal-regulated kinases （ERK） in hippocampus of the offspring rats of different genders. DESIGN ： A randomized and control animal experiment.SETTING： Department of Physiology and Pathophysiology, School of Medicine, Xi＇an Jiaotong University. MATERIALS ： The experiments were carried out in the Key Laboratory of Environment and Gene Related Diseases （Xi＇an Jiaotong University）, Ministry of Education between October 2005 and March 2006. Fifteen female and five male adult Sprague-Dawley rats were adopted. Female rats weighing 230-250 g and male rats weighing 280-350 g were used. METHODS： The virgin female rats were placed overnight with adult male rats （3：1） for mating. A total of twelve pregnant rats were randomly assigned to prenatal stress group （PNS group, n=6） and control group （n=6）. The pregnant rats of the PNS group were exposed to restraint stress on days 14-20 of pregnancy three times a day, 45 minutes for each time . The restraint device was a transparent plastic tube （6.8 cm in diameter） with air holes for breathing and closed end. The length could be adjusted to accommodate the size of the animals. To prevent habituation of animals to the daily procedure, restraint periods were randomly shifted within certain time periods （8：00-11：00, 11：00-14：00, and 16：00-19：00）. After birth, offsprings of all groups were culled to 8-10 litters in each group and housed in the same animal room, and kept together with their biologic mothers. The pregnant rats of the control group were left undisturbed. On day 21, after all the offspring were weaned, male and female pups were separated and housed four in each cage respectively until test at 30 days of age. At the end of postnatal day 30, one male and female offspring rats from the same dam were selected with a random choice and a total of 24 animals from 12 different dams were used. The experimental rats were sacrificed by decapitation under anesthesia. Bilateral hippocampal tissues were isolated and homogenized in cold condition. Alkaline carbonate buffer （BCA） method was used to detect the concentration of extracellular signal-regulated kinases （ERK）, then mixed with loading buffer, the constant voltage was 100 V. Finally, BCIP/NBT staining and electrDphoresis were performed, the absorbance （A） value for the bands was detected with the Bandscan analytical software, and the expression of ERK in hippocampus of offspring rats of different genders in each group was quantitatively analyzed. MAIN OUTCOME MEASURES： The level of ERK expression in hippocampus of offspring rats of different genders in each group was observed.RESULTS： All the 24 offspring rats were involved in the analysis of results. ① The staining results of ERP activity in the extract of brain tissue detected with Western blotting technique and specific antibody analysis showed that the ERP in hippocampus of offspring rats had two subtypes of ERK-1 and ERK-2, and the latter was the main type.② Standardized by the average A value in the control group, the quantitative data of the general A value of total ERK showed that the expression of ERK-2 in hippocampus of female offspring rats was obviously higher in the PNS group than in the control group （A value： 126±6.76,100±4.89,P〈 0.01）. ③The expression of ERK-2 had no obvious difference between the female and male offspring rats in the control group.④ The expression of ERK-2 in hippocampus of male offspring rats was a little higher in the PNS group than in the control group （A value： 104±6.27,102±5.48,P 〉 0.05）. CONCLUSION ： PNS significantly affects the increase of ERK expression in hippocampus of female offspring rats, but has no obvious influence on that of male ones.

Radioprotective effects of the expression of FLT3 ligand regulated by Egr-1 regulated element on radiation injury of SCID mice

Objective: In order to explore the radioprotective effects of the expression of hematopoietic growth factors regulated by radio-inducible promoter on radiation injury. Methods:The human FL (Flt3 ligand) cDNA and EGFP (enhanced green fluorescent protein) cDNA were linked together with IRES and then inserted into the eukaryotic expression vector pCI-Egr, which was constructed by substituting CMV promoter in pCIneo with the Egr-1 promoter (Egr-EF). The vector was transferred into human bone marrow stromal ...

Effect of sirolimus on the expression of integrin αυβ3 mRNA in cardiac allograft in rats

Objective To investigate the effect of sirolimus ( SRL) on the expression of integrin αυβ3 mRNA in vascular smooth muscle cells of cardiac allografts in rats, and the possible mechanism of SRL in the prevention of cardiac allograft vasculopathy. Methods Heterotopic heart transplantation models were established Hearts

Research on the effects of PIAS3 expression on the invasion of glioma TJ905 cells

Objective To investigate the function and possible mechanisms of PIAS3 expression on the invasion of TJ905 cells. Methods PIAS3 overexpression vectors were constructed and PIAS3 siRNA were chemically synthesized, which were separately transfected into TJ905 cells for upregulation or

Effect of different therapies of Chinese medicine on the expressions of c-Fos and c-Jun proteins in hippocampus of rats with post-stroke depression

BACKGROUND ： c-fos and c-jun, the important immediate early genes （IEG）, are regarded as the markers for the location and function of neuronal activity, as well as the third signal messengers, they couple the stress stimulation and the gene expression in neuron, and hippocampus is involved in the process of signal transmission after stress stimulation induced depression. OBJECTIVE： To observe the therapeutic effects of Bushen Yiqi （tonifying kidney to benefit qi）, Huoxue Huayu （promoting blood circulation to dissipate blood stasis） and Ditan Kaiqiao （eliminating phlegm for resuscitation） on the expressions of c-Fos and c-Jun proteins in hippocampus and spontaneous behaviors of rats with post-stroke depression （PSD）, and compare the results with those of fluoxetine, which is known to have definite effect on depression. DESIGN： A randomized controlled tna SETTING ： Zhejiang College of Traditional Chinese Medicine MATERIALS ： The trial was completed in Zhejiang College of Traditional Chinese Medicine from January to July in 2003. Fifty-six healthy adult Wistar male rats of clean grade, weighing （250±50） g, were randomly divided into 7 groups with 8 rats in each group： control group, model group, forced swimming group, Bushen Yiqi group; Huoxue Huayu, Ditan Kaiqiao group and fluoxetine group. The Bushen Yiqi Tang contained Renshen, Huangqi, Heshouwu, Gouqi, Shudi, etc., crude drugs 1 800 g/L. The Huoxue Huayu Tang contained Danshen, Chuanxiong, Chishao, Yujin, etc., crude drugs 3 600 g/L. The Ditan Kaiqiao Tang contained Banxia, Danxing, Changpu, Yuanzhi, etc., crude drug 1 000 g/b METHODS： ① Except the control group and forced swimming group, rats in the other groups were made into PSD models by deligating the bilateral common carotid artedes permanently. ② Rats in the control group, model group and forced swimming group were intragastncally perfused by saline （3 mL for each time）; those in the Bushen Yiqi group, Huoxue Huayu, Ditan Kaiqiao group and fluoxetine group were intragastncally perfused with Bushen Yiqi Tang （18 g/kg）, Huoxue Huayu Tang （9 g/kg）, Ditan Kaiqiao Tang （9 g/kg） and fluoxetine （2.5 mg/kg） respectively, once a day. ③ At 55 days after model establishment, rats in the forced swimming group were managed according to the Porsolt＇s method. They were placed in water for 15 minutes, and then taken out and dned, no moving-time within 5 minutes was recorded at drying and 24 hours after drying. ④ Measurement of spontaneous behaviors： Except the forced swimming group, the spontaneous behaviors and activities （including horizontal and vertical movements） of rats were observed with the Open-Field method at 28, 42 and 56 days after administration in the other groups. ⑤ The expressions of c-Fos and coJun proteins in hippocampus were determined with the immunohistochemical method, the relative sectional area ratio and average objective gray value of c-Fos and c-Jun positive cells in hip- pocampus were measured with the computerized image analytical system. MAIN OUTCOME MEASURES： The spontaneous behaviors of rats, the relative sectional area ratio and average objective gray value of c-Fos and c-Jun positive cells in hippocampus were observed. RESULTS： Of the 56 rats, 1 died in the forced swimming group, and finally 55 rats were involved in the analysis of results. ① Results of spontaneous activities： At 28 days, the times of crossing movements were obviously fewer in the model group and fluoxetine group [（69.00±37.01）, （98.11 ±36.68） times/3 minutes] than in the control group [（128.44±16.85） times/3 minutes, P 〈 0.01, 0.05], but those in the Bushen Yiqi group, Huoxue Quyu group and Ditan Kaiqiao group had no obvious differences as compared with those in the control group （P 〉 0.05）. At 42 and 56 days, the times of crossing movements were obviously more in the Bushen Yiqi group, Huoxue Quyu group and Ditan Kaiqiao group [（106.44±31.24）, （117.20±23.95）, （134.80±28.18）, （136.36±40.95） times/3 minutes; （117.33±35.91）, （129.60 ±23.78）, （131.90 ±26.81）, （136.09±28.34） times/3 minutes] than in the model group [（64.00±17.51）, （72.86±20.68） times/3 minutes, P 〈 0.01]. The times of rearing movements had no obvious differences among the groups for the three times （P 〉 0.05）. ② The no moving-time within 5 minutes 24 hours after drying was obviously longer than that at drying in the forced swimming group. ③ The average objective gray values of c-Fos positive cells were not obviously different in the Bushen Yiqi group and Ditan Kaiqiao group from the control group （P 〉 0.05）, but lower in the model group than in the control group （69.84±9.82, 75.78±5.89, P 〈 0.01）, and higher in the forced swimming group than in the control group （85.97±10.99, P 〈 0.01）; all higher in the fluoxetine group, Bushen Yiqi group, Huoxue Quyu group and Ditan Kaiqiao group than in the model group （81.27±10.73, 74.04±8.34, 83.29±9.89, 70.14±4.92, P 〈 0.05-0.01）. The average objective gray values of c-Jun positive cells were obviously lower in the Bushen Yiqi group than in the control group （68.11 ±6.89, 79.58±5.86, P 〈 0.01）, but all higher in the other groups than in the control group （84.68±7.15, 81.34 ±8.36, 97.51±10.55, 85.68±9.25, 86.19±10.98, P 〈 0.05-0.01）; Those were obviously higher in the fluoxetine group, Huoxue Quyu group and Ditan Kaiqiao group than in the model group （P 〈 0.05-0.01 ）, lower in the Bushen Yiqi group than in the model group （P 〈 0.05）, all obviously lower in the Bushen Yiqi group, Huoxue Quyu group and Ditan Kaiqiao group than in the fluoxetine group （P 〈 0.01）. The relative sectional area ratios of c-Fos and c-Jun positive cells had no obvious differences among the groups （P 〉 0.05）. CONCLUSION ： The methods of Bushen Yiqi, Huoxue Quyu and Ditan Kaiqiao can effectively treat PSD in rats, and the results were equivalent with those of fluoxetine, the actions of the above-mentioned drugs may correlated with their regulation to c-Fos and c-Jun expressions in hippocampus. PSD animal models can be successfully established by both permanent deligation of bilateral common carotid arteries and forced swimming, and the models induced by the former has similar basic cerebrovascular lesions as human stroke in clinic.

Effect of FTY720 on RhoA expression after acute spinal cord injury in rats

Objective To investigate the effect of fingolimod (FTY720)on RhoA expression after spinal cord injury (SCI)in rats,and explore the possible mechanism of FTY720in the treatment of SCI.Methods A rat model of acute SCI was established with a

Effects of electrostimulation and administration of succinylcholine on the expression of Fos protein in mesencephalic periaqueductal gray matter of rats after simulated weightlessness

BACKGROUND： Expression of Fos in neurons of periaqueductal gray （PAG） is used to reflect the excitability. However, changes of expression of Fos in neurons of PAG are caused by injured electrostimulation after simulated weightlessness, and the relationship between pretreatment and injection of succinylcholine has not been determined yet. OBJECTIVE ： To investigate the changes of expression of Fos in PAG induced by injured electrostimulation pretreatment and injection of succinylcholine at 2 weeks after simulated weightlessness.DESIGN： Observational and controlled animal study.SETTING： Department of Physiology, Medical School, Xi＇an Jiaotong University; Key Laboratory of Environment and Genes Related to Diseases, Ministry of Education. MATERIALS： A total of 24 adult female SD rats, of clean grade and weighing 180-220 g, were selected in this study. METHODS： The experiment was completed in the Experimental Animal Center of Xi＇an Jiaotong University.① All rats were randomly divided into 2 groups according to body mass： simulated weightlessness group and control group with 12 in each group. And then, each group was also divided into 3 subgroups： electrostimulation group, succinylcholine-pretreatment group and succinylcholine-injection group with 4 in each subgroup. ②The model of weightlessness was simulated by tail-suspended female rats, which were described and modified by Cheng Jie. Rats in normal control group were given the same interventions as simulated weightlessness group except for tail-suspended. ③ Experimental method： The rats in electrostimulation group were given nociceptive stimulus by a pair of subcutaneous electrodes inserted into 1 and 5 claw of left hindlimb. The stimulus （current： 10 mA; duration： 1 ms; interval： 1 s） lasted for 30 minutes. The rats in succinylcholine-pretreatment group received stimulus after intravenous administration of succinylcholine, rats in succinylcholine-injection group were not given stimulus, just received succinylcholine. ④ All rats were perfused and fixed after 2 hours from the end of stimulation. The brains were removed, and serial frozen sections of midbrain were stained using immunocytochemical method, observed and taken photos under light-microscope. The number and morphological characters of Fos-immunoreactive （Fos-IR） neurons in ventrolateral part of PAG were investigated. MAIN OUTCOME MEASURES： The alterations in number and morphological characters of Fos-IR neurons in ventrolateral PAG of all rats.RESULTS： A total of 24 rats were involved in the final analysis. ① The morphological changes of Fos-IR neurons： The expressions of Fos in ventrolateral part of PAG were observed in both control and simulated weightlessness groups rats after being given nociceptive stimulus. As compared with control group, Fos-IR neurons in simulated weightlessness group were dyed lightly, cellular integrity was impaired, and cellular verge was unclear. ② The numbers of Fos-IR neurons： In control group, the numbers of Fos-IR neurons in ventrolateral part of PAG in simulated weightlessness group were obviously lower than succinylcholine-pretreatment group, but obviously higher than succinylcholine-injection group （46.94±3.38, 71.06±8.96 and 35.04±4.62, respectively, P 〈 0.05）. In 14-day simulated weightlessness group, the numbers of Fos-IR neurons in electrostimulation group were also obviously lower than succinylcholine-pretreatment group, and obviously higher than succinylcholine-injection group （27.77±3.27, 32.91±2.99 and 11.75±1.00, respectively, P 〈 0.05）. The numbers of Fos-IR neurons in all subgroups in control group were obviously higher than those subgroups in simulated weightlessness group. Compared with electrostimulation group, the percentage of expression of Fos in ventrolateral part of PAG responsed to nociceptive stimulus after administration of succinylcholine （SCH） was increased to 51.83% in control group and 18.51% in simulated weightlessness group.CONCLUSION ：① The expression of Fos in neurons in ventrolateral part of PAG were increased by the pretreatment of SCH before nociceptive stimulus.② Nociceptive stimulus could increase the expression of Fos in neurons in ventrolateral part of PAG. ③ The numbers of Fos-IR neurons in ventrolateral part of PAG were decreased obviously after 2-week simulated weightlessness.