Wideband frequency-dependent dielectric properties of rat tissues exposed to low-intensity focused ultrasound in the microwave frequency range

Tissue dielectric properties can vary upon the incident of an acoustic wave.The goal of this study is to quantify this change due to the acoustoelectric effect(AE),and to obtain the frequency-dependent dielectric properties of tissues exposed to low-intensity focused ultrasound(LIFU).The dielectric properties of the blood,brain,chest muscle,heart,kidney,leg muscle,liver,lung,pancreas,and spleen of rats were measured by an open-ended coaxial probe method.The acoustic intensity of LIFU focus was 2.97 MPa(67.6 W/cm^(2)),3.95 MPa(120 W/cm^(2)),and 5.17 MPa(204 W/cm^(2)),respectively,and the measurement frequency band was 0.1–7.08 GHz.The measurement results show that with the LIFU modulation,the conductivity and dielectric constant decreased in the high-frequency band,and on the contrary,they increased in the lowfrequency band,and the larger the acoustic intensity was,the more obvious the phenomenon was.This work contributes to a better understanding of the mechanisms by which ultrasound acts on the dielectric properties of biological tissues.It is expected that the findings from this study will provide a basis that the response of tissue to LIFU modulation can be monitored by noninvasive techniques such as microwave-induced thermoacoustic imaging(MTI)and microwave imaging,present a new idea for improving the endogenous contrast between different biological tissues in MTI and acoustoelectric imaging,and possibly lead to the development of a new imaging method based on the relaxation time of tissue after LIFU modulation.

Simultaneous determination mercury species of Su-He-Xiang-Wan in rat tissues by HPLC-CVG-AFS

A simple and sensitive high performance liquid chromatography-chemical vapour generation-atom fluorescent spectrometry （HPLC-CVG-AFS） method was developed and validated for simultaneous determination mercury species in Su-He-Xiang-Wan （SHXW） and in tissues of rats, respectively. The species of mercury were separated by a Venusil MP-C 18 （5μm, 150 mm×4.6 ram） column with the optimized mobile phase containing 5% （w/v） acetonitrile, 0.01 mol/L L-cysteine and 0.06 moL/L ammonium acetate. The tissues of rats were freeze-dried after giving the medicine for 10 d, and then added into the solution containing 10% （w/v） HC1, 1% （w/v） sulfocarbamide and 0.15% （w/v） KC1 for increasing extraction rate. The resolutions of Hg2＋, MeHg and EtHg were 1.5 and 2.9, respectively. The detection limits of Hg2＋, MeHg and EtHg were 2.0, 1.0 and 0.9 ng/mL, respectively. The relative standard deviation （RSD） of inter- and intra-day precisions ranged from 1.56% to 2.86%. The recovery rates of three different adding level were 87%-101% （n=6）, and the RSDs were smaller than 8.2%. The results show that no MeHg and EtHg were detected in rat tissues. Only soluble mercury （Hg2＋） was determined for the mercury species of SHXW in rat tissues.

Pathological Changes in the Sinoatrial Node Tissues of Rats Caused by Pulsed Microwave Exposure

To observe microwave induced dynamic pathological changes in the sinus nodes, wistar rats were exposed to 0, 5, 10, 50 mW/cm^2 microwave. In 10 and 50 mW/cm^2 groups, disorganized sinoatrial node cells, cell swelling, cytoplasmic condensation, nuclear pyknosis, and anachromasis, swollen, and empty mitochondria, and blurred and focally dissolved myofibrils could be detected from 1 to 28 d, while reduced parenchymal cells, increased collagen fibers, and extracellular matrix remodeling of interstitial cells were observed from 6 to 12 months. In conclusion, 10 and 50 mW/cm^2 microwave could cause structural damages in the sinoatrial node and extracellular matrix remodeling in rats.

Comparison of T-2 Toxin and HT-2 Toxin Distributed in the Skeletal System with That in Other Tissues of Rats by Acute Toxicity Test

Twelve healthy rats were divided into the T-2 toxin group receiving gavage of 1 mg/kg T-2 toxin and the control group receiving gavage of normal saline. Total relative concentrations of T-2 toxin and HT-2 toxin in the skeletal system（thighbone, knee joints, and costal cartilage） were significantly higher than those in the heart, liver, and kidneys（P 〈 0.05）. The relative concentrations of T-2 toxin and HT-2 toxin in the skeletal system（thighbone and costal cartilage） were also significantly higher than those in the heart, liver, and kidneys. The rats administered T-2 toxin showed rapid metabolism compared with that in rats administered HT-2 toxin, and the metabolic conversion rates in the different tissues were 68.20%-90.70%.

Changes of learning and memory ability associated with neuronal nitric oxide synthase in brain tissues of rats with acute alcoholism

BACKGROUD： Ethanol can influence neural development and the ability of leaming and memory, but its mechanism of the neural toxicity is not clear till now. Endogenous nitric oxide （NO） as a gaseous messenger is proved to play an important role in the formation of synaptic plasticity, transference of neuronal information and the neural development, but excessive nitro oxide can result in neurotoxicity. OBJECTIVE ： To observe the effects of acute alcoholism on the learning and memory ability and the content of neuronal nitric oxide synthase （nNOS） in brain tissue of rats. DESIGN ： A randomized controlled animal experiment. SETTING ： Department of Physiology, Xinxiang Medical College MATERIALS： Eighteen male clean-degree SD rats of 18-22 weeks were raised adaptively for 2 days, and then randomly divided into control group （n = 8） and experimental group （n = 10）. The nNOS immunohistochemical reagent was provided by Beijing Zhongshan Golden Bridge Biotechnology Co.,Ltd. Y-maze was produced by Suixi Zhenghua Apparatus Plant. METHODS ： The experiment was carded out in the laboratory of the Department of Physiology, Xinxiang Medical College from June to October in 2005. ① Rats in the experimental group were intraperitoneally injected with ethanol （2.5 g/kg） which was dissolved in normal saline （20%）. The loss of righting reflex and ataxia within 5 minutes indicated the successful model. Whereas rats in the control group were given saline of the same volume. ② Examinations of learning and memory ability： The Y-maze tests for learning and memory ability were performed at 6 hours after the models establishment. The rats were put into the Y-maze separately. The test was performed in a quiet and dark room. There was a lamp at the end of each of three pathways in Y-maze and the base of maze had electric net. All the lamps of the three pathways were turned on for 3 minutes and then turned off. One lamp was turned on randomly, and the other two delayed automatically. In 5 seconds after alternation, pulsating electric current presented in the base of unsafe area to stimulate rat＇s feet to run to the safe area. The lighting lasted for 15 seconds as one test. Running from unsafe area to safe area at one time in 10 seconds was justified as successful. Such test was repeated for 10 times for each rat and the successful frequency was recorded. The qualified standard of maze test was that the rat ardved in the safe area g times during 10 experiments. The number of trainings for the qualified standard was used to represent the result of spatial learning. ③ Determination of the content of nNOS in brain tissue： After the Y-maze test, the rats were anaesthetized, and blood was let from the incision on right auricle, transcardially perfused via the left ventricle with about 200 mL saline, then fixed by perfusion of 40 g/L paraformaldehyde. Hippocampal CA1 region, corpus striatum and cerebellum were taken to prepare serial freezing coronal sections. The nNOS contents in the brain regions were determined with the immunohistochemical methods to reflect the changes of nitdc oxide in brain tissue. MAIN OUTCOME MEASURES ： The changes of learning and memory ability and the changes of the nNOS contents in the brain tissue of rats with acute alcoholism were observed. RESULTS ： One rat in the experimental group was excluded due to its slow reaction to electdc stimulation in the Y-maze test, and the other 17 rats were involved in the analysis of results. ① The training times to reach qualifying standards of Y-maze in the expedmental group was more than that in the control group [（34.33 ±13.04）, （27.50±8.79） times, P〈 0.05]. ② Forms and numbers of nNOS positive neurons in brain tissue： It could be observed under light microscope that in the hippocampal CA1 region, there were fewer nNOS positive neurons, which were lightly stained, and the processes were not clear enough; But the numbers of the positive neurons which were deeply stained as huffy were obviously increased in the experimental group, the cell body and cyloplasm of process were evenly stained, but the nucleus was not stained. The nNOS positive neurons in corpus stdatum had similar forms and size in the experimental group and control group. The form of the nNOS positive neurons in cerebellum were similar between the two groups. The numbers of nNOS positive neurons in hippocampal CA1 region and corpus striatum in the expedmental group [（18.22±7.47）, （11.38±5.00） cells/high power field] were obviously higher than those in the control group [（10.15±4.24）, （6.15±3.69） cells/high power field. The number of nNOS positive neurons in cerebellum had no significant difference between the two groups [（49.56±18.84）, （44.43±15.42） cells/high power field, P〉 0.05]. CONCLUSION ： Acute alcoholism may impair learning and memory ability, and nitric oxide may be involved in mediating the neurotoxic role of ethanol.

In vitro release of 1,3-bis(2-chloroethyl)-1-nitrosourea sustained-release microspheres and the distribution in rat brain tissues

BACKGROUND: The implantation of released chemotherapeutic drugs, which takes biodegradable polymer as vector, into the tumor site can get high concentration and release the drug for a long time, it can directly act on the tumor cells, and reduce the general toxicity. OBJECTIVE: To explore the in vitro and in vivo course of 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU) sustained-release from BCNU-loaded polylactide (PLA) microspheres (MS) and location in rat brain tissue. DESIGN: A repetitive measurement. SETTING:Central Pharmacy, General Hospital of Chinese People’s Armed Police Forces. MATERIALS: Thirty male SD rats were used. PLA (Mr5000, batch number: KSL8377) was produced by Wako Pure Chemical Inc.,Ltd. (Japan); BCNU (batch number: 021121) by Tianjin Jinyao Amino Acid Co., Ltd.; BCNU-PLA-MS was prepared by the method of solvent evaporation and pressed into tablets (10 mg/tablet). High-performance liquid chromatography (HPLC) Agilent 1100 (USA); LS9800 liquid-scintillation radiometric apparatus (Beckman). Chromatographic conditions: Elite Hypersil ODS2 C18 chromatographic column (5 μm, 4.6 mm×150 mm); Mobile phase: methanol: water (50:50), flow rate was 1.0 mL per minute, wave length of ultraviolet detection was 237 nm, and the inlet amount of samples was 10 μL. METHODS: The experiments were carried out in the experimental animal center of the General Hospital of Chinese Armed Police from May 2004 to July 2005. ① In vitro BCNU-PLA-MS release test: BCNU-PLA-MS was prepared by the method of solvent evaporation, then placed in 0.1 mol/L phosphate buffered solution (PBS, pH 7.4, 37 ℃), part of MS were taken out at 1, 2, 3, 7, 10 and 15 days respectively, and the rest amount of BCNU in MS was determined by HPLC, then the curve of BCNU-PLA-MS release was drawn. ②In vivo BCNU-PLA-MS release and distribution test: The rats were anesthetized, then BCNU-PLA-MS were implanted to the site 1 mm inferior to the cortex of frontal lobe. Five rats were killed postoperatively at 4 hours, 1, 2, 3, 7 and 15 days, the residual MS was removed from the brain tissue. The rest amount of BCNU was determined with HLPC, and the curve of BCNU-PLA-MS release was drawn as compared with the amount of BCNU in the implanted tablets. Besides, brain tissues (1 g) at the implanted side and the contralateral one were obtained respectively, blood sample (0.5 mL) was also collected, 3H-BCNU was counted radioactively in radioactive liquid flash solution. The distributions of BCNU-PLA-MS in normal rat brain tissue and serum were detected. The analysis of variance was applied to compare the intergroup differences of the measurement data. MAIN OUTCOME MEASURES: ① Characteristics of BCNU-PLA-MS release in phosphate buffered solution (PBS) and rat brain tissue; ② Distributions of BCNU-PLA-MS in normal rat brain tissue and serum. RESULTS: ① Release of BCNU-PLA-MS in PBS and rat brain tissue: The BCNU released from BCNU-PLA-MS could be sustained for over 2 weeks both in PBS and brain tissue. In PBS, the released rate of BCNU was over 15% at 24 hours, nearly 50% at 72 hours and over 90% at 15 days. In brain tissue, the released rate was 8% at 4 hours, 16% at 24 hours, 60% at 72 hours, respectively, and BCNU could be sustained released for over 15 days. ② Distributions of BCNU-PLA-MS in normal rat brain tissue and serum: The concentrations of BCNU in the ipsilateral brain tissue were 6 to 70 times higher than those in the contralateral one. The concentrations of BCNU in the ipsilateral brain tissue were obviously higher than those in serum and contralateral brain tissue (F =103.47, P < 0.01). CONCLUSION: BCNU-PLA-MS can increase the drug concentration in targeted brain tissue, decrease that in the non-targeted brain tissue, reduce general toxic and side effects, and have good releasing function.

Electro-acupuncture for STAT3 expression and nuclear translocation in hippocampal tissues of rats following cerebral ischemia/reperfusion

BACKGROUND: It has been found in recent years that STAT3 widely distributes in nervous system, including hippocampal CA1-3 region, dentate gyrus and cerebral neocortex, etc. Ischemic brain injury can cause the release of some cytokines and growth factors, while electro-acupuncture may have multi-level, multi-channel and multi-target protective and interventional effects on ischemic brain injury. OBJECTIVE: To observe the effects of electro-acupuncture on STAT3 expression and nuclear translocation in hippocampal CA1 region of rat models of brain ischemia/reperfusion. DESIGN: Randomized and controlled observation. SETTING: Staff Room of Acupuncture and Moxibustion, Department of Acupuncture and Bone Injury, Hubei College of Traditional Chinese Medicine; Tongji Medical College, Huazhong University of Science and Technology. MATERIALS: Seventy-two healthy SD rats, of clean degree and either gender, weighing (200±20) g, were provided by the Experimental Animal Center of Hubei College of Traditional Chinese Medicine. STAT3 monoclonal antibody was purchased from Santa Cruz Company, USA, and G-6805 electro-acupuncture instrument was purchased from Shanghai Medical Electronic Instruments Factory. METHODS: This experiment was carried out in the comprehensive laboratory of Department of Acupuncture and Bone Injury, Hubei College of Traditional Chinese Medicine between September 2005 and February 2006. Seventy-two rats were randomly divided into 4 groups: ① control group(n =6): Untouched. ② Sham-operation group (n =18): Artery was isolated, but without inserting thread bolt.③ Model group (n =24): Rat models of local brain ischemia/reperfusion were established with modified suture occlusion. ④Electro-acupuncture group (n =24): Dazhui and bilateral Neiguan points were selected for electro-acupuncture treatment. No. 28 acupuncture needle of 3.33 cm was used in the treatment. A G-6085 electro-acupuncture instrument with continuous wave, frequency of 120 times/min, intensity of 1 mA, 30 min/time, was used. Acupuncture was conducted firstly at ischemia/reperfusion 3 hours, then once every 12 hours. STAT3 positive nuclear translocation in hippocampal CA1 region of rats was observed with immunohistochemical method at 24, 48 and 72 hours after brain ishcemia/reperfusion, and then STAT3 positive cells were counted. MAIN OUTCOME MEASURES: STAT3 positive cells and nuclear translocation in hippocampal CA1 region of rats in each group. RESULTS: All the 72 rats were involved in the result analysis. ①In the control group and sham-operation group, STAT3 positive cells with light cytoplasm and nucleus were decreased , and nuclear translocation was not found. ② In the model group, STAT3 positive cells were mostly found in the cytoplasm of the hippocampal CA1 region at the ischemic side of rats after ischemia/reperfusion 24 hours. They were significantly more than those in the sham-operation group and control group [(18.00±2.68), (9.00±1.35), (8.00±1.22) cells/ mm2, P < 0.01], but cells with nuclear reaction were fewer; At ischemia/reperfusion 48 and 72 hours, STAT3 positive cells were increased, and they were significantly more than those of sham-operation group [(25.00±3.23), (35.00±3.52) cells/mm2, (13.00±1.93), (12.00±1.24) cells/mm2, P < 0.01]. Positive cells with nuclear reaction were found dark-stained. ③At ischemia/reperfusion 24, 48 and 72 hours, STAT3 positive cells were strongly expressed in hippocampal CA1 region at ischemic side of rats of electro-acupuncture group, and they were significantly more than those of model group [(25±3.52), (50±6.31), (75±8.09) cells/mm2, P < 0.01]. STAT3 positive cells were gradually enhanced with time, and considerable STAT3 nuclear positive reaction cells were found. CONCLUSION: Electro-acupuncture can activate STAT3 protein expression in hippocampal tissue of rats with local brain ischemia/reperfusion, promote STAT3 nuclear translocation and function its neuroprotective effect.

Changes in hemeoxygenase-1 and superoxide dismutase in the peri-hematomal brain tissues of rats following intracerebral hemorrhage

BACKGROUND: The mechanism of intracerebral hemorrhage (ICH)-induced hemorrhagic brain injury is very complicated, involving the position-occupying effect of cephalophyma, ischemic factors, the toxic effect of hematoma components, the destruction of blood-brain barrier, etc. The expression and effect of hemeoxygenase-1 (HO-1) in the cerebrovascular disease has been paid close attention. OBJECTIVE: To observe the expression of HO-1 and change of superoxide dismutase (SOD) in the peri-hematomal brain tissue of rats following ICH. DESIGN: Randomized controlled animal experiment. SETTING: Department of Neurology, Yijishan Hospital Affiliated to Wannan Medical College. MATERIALS: Forty healthy male SD rats, of clean grade, weighing from 250 to 300 g, were provided by Qinglongshan Animal Farm of Nanjing. The involved 40 rats were randomized into sham-operation group (n =5) and ICH group (n =35), and ICH group was divided into 7 subgroups with 5 rats in each: ICH 6, 12, 24, 48, 72, 100 and 168 hours groups. Rabbit anti-rat HO-1 immunohistochemial kit ( Boster Co., Ltd., Wuhan) and SOD kit (Jiancheng Bioengineering Institute, Nanjing)were used in this experiment. METHODS: This experiment was carried out in the Department of Neurology, Yijishan Hospital Affiliated to Wannan Medical College Between April and July 2005. In the ICH group: Autologous blood of rats was injected into the head of caudate nucleus to create ICH animal models. In the sham-operation group, the same amount of normal saline was injected into the head of caudate nucleus of rats. The brains of rats in each group were harvested at different time points. The hematoma-side brain tissue was cut open in the coronal plane taking hematomal region as center, and the posterior part was fixed with 100 g/L neutral formaldehyde. 100 mg brain tissue was taken from anterior part. The number of positive cells in HO-1 and SOD activity in peri-hematomal brain tissue at different time after ICH were detected by immunohistochemical method and xanthine oxidation method respectively. MAIN OUTCOME MEASURES: ① The expression of HO-1 in the peri-hematomal brain tissue of rats in two groups following ICH.② The expression of SOD activity in the peri-hematomal brain tissue of rats in two groups following ICH. RESULTS: ①The number of HO-1 positive cells in the peri-hematomal brain tissue of rats in two groups following ICH 6, 12, 24, 48, 72, 120 and 168 hours was (11.03±2.01),(16.47±2.98),(25.50±5.65),(51.57±7.05),(47.33±4.73),(26.57±5.12),(7.63±2.17) cells/high-fold visual field , respectively; The number of HO-1 positive cells in the ICH 12-120 hours groups was significantly higher than that of sham-operation group [(6.07±1.85)cells/high-fold visual field, P < 0.01]; The HO-1 positive cells were the most in the ICH 48 hours group and were still expressed a little in the ICH 168 hours group. ② The SOD in the brain tissue of rats at ICH 6, 12, 24, 48, 72, 120 and 168 hours was (404.46±8.14),(396.84±10.97),(387.74±5.32),(356.21±9.27),(307.95±10.15),(357.48±11.28) and (402.98±7.23) kNU/g, respectively; The SOD activity of ICH 12 to 120 hours groups was significantly lower than that of sham-operation group [(415.47±11.44) kNU/g,P < 0.01], and that of ICH 72 hours group was the lowest. There was no significant difference of SOD activity between ICH 168 hours group and sham-operation group (P > 0.05). CONCLUSION: Following ICH, the expression of HO-1 in peri-hematomal brain tissue of rats in two groups is obviously increased, but the antioxidant ability of brain tissue is decreased. The changes of both maybe play an important role in the formation of ICH-induced hemorrhagic brain injury.

Effect of nitric oxide synthase inhibitor N^G-nitro-L-arginine on the content of amino acid in ischemic brain tissues of rats

BACKGROUND： Nitric oxide synthase （NOS） inhibrtors have been widely used to investigate the role of NO on cerebral ischemic injury, but the results are controversial. Moreover, it has been considered to aggravate the ischemic neuronal damage with the release of excessively excitatory amino acids （EAA） during cerebral ischemia. On the other hand, some inhibitory amino acid is suggested to be important for the neuronal protection against ischemic brain damage. Our study has recently showed that treatment with the NOS inhibitor NG-nitro-L-arginine （L-NA） reduced focal cerebral ischemic damage. The effect of L-NA on the contents of excitatory and inhibitory amino acid in the rat brain following cerebral ischemia is still unclear. OBJECTIVE： By evaluating the effect of NOS inhibitor, L-NA on the contents of aspartate, glutamate, glycine and γ-aminobutyric acid （GABA） in striatum, hippocampus and cortex in the rat brain following cerebral ischemia respectively, to investigate the beneficial effect of L-NA on cerebral ischemic injury and the possible mechanism. DESIGN： A randomized and controlled experiment SETTING ： Department of Pharmacology, Hebei Academy of Medical Sciences MATERIALS： A total of 42 male healthy SD rats （grade Ⅱ, weighting 250-300 g） were provided by the Experimental Animal Center of Hebei Province （Certification： 04036）. Aspartate, glutamate, glycine, GABA, L-NA and 2,3,5-triphenyltetrazolium chloride （TTC） were obtained from Sigma Chemicals Co, St Louis, MO, USA. HPLC-ultraviolet detector system consisted of Agilent 1100 HPLC. METHODS： The experiment was carried out in Department of Pharmrcology, Hebei Academy of Medical Sciences from June 2005 to June 2006. Rats were randomly divided into three groups： sham-operated group （n = 6）, ischemic group （n = 18）, L-NA group （n = 18）. The model of focal cerebral ischemia in rat was prepared with intraluminal line occlusion methods. In sham-operated rats, the external carotid artery was surgically prepared, but the filament was not inserted. Each group was further divided into 3 subgroups （n = 6 for each）： drugs were administrated at 2, 6 and 12 hours after the middle cerebral artery occlusion （MCAO） respectively. L-NA （20 mg/kg, ip） was administrated, twice a day, for 3 consecutive days. Same volume of normal saline was administrated in ischemic and sham operation groups. The changes of infarcted volume and the contents of amino acids were respectively assayed. Image analysis software was used for the measurement of the infarcted area. The results were expressed as a percentage of the infarcted volume of cerebral/volume of whole brain （IV%） in order to control for edema formation. The contents of aspartate, glutamate, glycine and GABA in striatum, hippocampus and cortex in the rat brain following cerebral ischemia were respectively measured by HPLC method. All data were analyzed with one-way ANOVA and Dunnett＇s test. MAIN OUTCOME MEASURES： （1） The volume of cerebral infarction; （2） The contents of aspartate, glutamate glycine and GABA in brain tissue after cerebral ischemia. RESULTS ： All 42 rats were involved in the final analysis. （1） Infarcted volume： Volume was 0 in sham-operated group. When L-NA was administrated at 2 and 6 hours after MCAO, the infarcted volume was （20.13±3.59）% and （23.12±5.84）% in L-NA group, which was not similar to that in ischemic group [（22.10±3.98）%, （25.38± 5.37）%, P〉 0.05]. However, the infarcted volume was markedly decreased compared with that of ischemic group when L-NA was administrated at 12 hours after MCAO [（26.11±3.55）% and （37.15±3.58）%, P 〈 0.01]. Changes of amino acid content： At 2 and 6 hours after ischemia, the contents of aspartate, glutamate, glycine and GABA in striatum, hippocampus and cortex in ischemic group were significantly increased compared with those in sham-operated group （ P〈 0.05-0.01）. However, contents in L-NA group were similar to those in ischemic group （P 〉 0.05）. At 12 hours after ischemia, the contents of aspartate [（0.21 ±0.06）, （0.36±0.05）, （0.29±0.12） mg/g] and glutamate [（0.55±0.06）, （0.78±0.10）, （0.52±0.10） mg/g] in striatum, hippocampus and cortex in L-NA group were significantly decreased compared with those in ischemic group [（0.49±0.17）, （0.63± 0.03）, （0.51±0.15） mg/g; （0.98±0.30）, （1.15±0.15）, （0.93±0.15） mg/g, P〈 0.05-0.01]. Glycine in hippocampus was （0.40±0.07） mg/g, which was higher than that in ischemic group [（0.21±0.07） mg/g, P 〈 0.05]. GABA in striatum, hippocampus and cortex was （0.93±0.10）, （0.62±0.12） and （0.81 ±0.10） mg/g, respectively, which was higher than that in ischemic group [（0.60±0.08）, （0.37±0.17）, （0.59±0.10） mg/g, P 〈 0.05-0.01]. CONCLUSION ： It may be concluded that L-NA have beneficial effect on ischemic cerebral injury in ischemic later stage in rats. The possible mechanism is that L-NA can decrease the contents of aspartate and glutamate, increase the contents of glycine and GABA.

Effect of Acetaldehyde on Oxidoreductases in Tissues of Rats at Different Ages

The toxicity of acetaldehyde and age related changes on oxidoreductases in the liver,brain, kidney, and musele of female albino rats (Wistar strain) were studied. The specific activities of lactate [LDH], isocitrate [ICDH (NAD/NADP)], succinate [SDH], malate [MDH], glutamate [GDH] and glucose-6-Phosphate [G-6-PDH] dehydrogenases were signifieantly inereased as a function of age. However, acetaldehyde treatment significantly inhibited oxidoreductases in the tissues of 21, 90 and 180 day old rats. Liver enzymes of young (21 days) rats exhibited greater sensitivity to acetaldehyde toxicity. Similar inhibition of oxidoreductases in brain and kidney of adult (180 days) rats treated with acetaldehyde was observed. LDH and GDH as compared to other enzymes studied showed higher susceptibility to acetaldehyde toxicity. The differential sensitivity of tissues and inhibition of oxidoreductases by acetaldehyde as a function of age could be attributed to hypoxic conditions, energy crisis, and mitochondrial structural changes. The results suggest that acetaldehyde affects oxidation of glucose via HMP shuni pathway, glycolytic pathway and Krebs cycle resulting in the impairment of carbohydrate metabolism

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.

Distribution in tissue and excretion in urine and feces of swertisin after intravenous administration to rats

Swertisin contents in rat urine,feces and tissues were determined by reversed-phase high-performance liquid chromatography（RP-HPLC） method.Chromatographic separations were performed on a C18 column with acetonitrile-water（23：77,v/v） as the mobile phase.The calibration curves were linear over the ranges of 0.175-35.0μg/mL for rat urine,0.5-60.0μg/mL for rat feces,and 0.014 to 53.0μg/mL for all tissues.The inter-and intra-day precisions and accuracy for all measured samples were satisfactory.The fully validated method was applied for tissue distribution and excretion of swertisin in rat urine and bile after intravenous administration.The maximum level of swertisin was found in kidney,which reached 83.87± 6.36μg/g.In rat heart,swertisin was hardly detected under used experimental conditions.Swetisin level in liver,kidney,stomach,smooth muscle and skeletal muscle continued to decrease from 5 to 60min.Swertisin showed increasing tendency in intestine,spleen and testis tissues at scheduled time points.Detectable swertisin was found in brain and lung tissue.Totally 11.9% swertisin dose was cumulatively excreted from urine in 60h after intravenous administration.There was small amount of swertisin in rat feces and the cumulative excretion level reached 4.59% of intravenous dose in 60h.

Expression of Peroxiredoxins and Pulmonary Surfactant Protein A Induced by Silica in Rat Lung Tissue

Silicosis is one of the most serious occupational diseases in China and dates back to centuries ago. In this study, we successfully established a rat model of silicosis by intratracheal silica injection for 28 days and determined hydroxyproline levels to evaluate collagen metabolism in lung homogenates. Oxidative stress status was evaluated by detecting catalase and glutathione peroxidase activities.

THE LOCALIZATION OF ADRENOMEDULLIN IN RAT KIDNEY TISSUE AND ITS INHIBITORY EFFECT ON THE GROWTH OF CULTURED RAT MESANGIAL CELLS

OBJECTIVE: To observe the localization of adrenomedullin (AM) in rat kidney tissue and its inhibitory effect on the growth of cultured rat mesangial cells (MsC). METHODS: A monoclonal antibody against AM developed by our laboratory was used to detect the localization of AM protein in rat kidney tissue by avidin-biotin complex immunohistochemistry. The expressions of AM and its receptor CRLR mRNA on cultured glomerular epithelial cells (GEC) and MsC were investigated by Northern blot assay, and the possible effect of AM secreted by GEC on MsC proliferation was observed using [3H]thymidine incorporation as an index. RESULTS: A specific monoclonal antibody against AM was succesfully developed. AM was immunohistochemically localized mainly in glomeruli (GEC and endothelial cells), some cortical proximal tubules, medullary collecting duct cells, interstitial cells, vascular smooth muscle cells and endothelial cells. Northern blot assay showed that AM mRNA was expressed only on cultured GEC, but not on MsC, however, AM receptor CRLR mRNA was only expressed on MsC. GEC conditioned medium containing AM can inhibit MsC growth and AM receptor blocker CGRP8-37 may partially decreased this inhibitory effect. CONCLUSION: AM produced by GEC inhibits the proliferation of MsC, which suggests that AM as an important regulator is involved in glomerular normal physiological functions and pathologic processes.

A NEW METHOD OF ISOLATING KUPFFER CELLS FROM BIOPSY TISSUE OF RAT LIVER

The isolation of a high yield and purity of Kupffer cells has been reported in detail.1 This paper reports into the research about isolation Kupffer cells from biopsy tissue of liver. This method includes 5 important steps: (1) take fresh liver tissue, and mince with scissors. (2) spin at low speed to wash off red blood cells. (3) digest in collagenase for suitable time. (4) isolate Kupffer cells on a percoll density gradient. (5) cell charaterization was observed by N.S.E stain and peroxidatic activity with lumino-meter measurement and phagocytosis with latex beads.2.3

Diazinon Toxicokinetics, Tissue Distribution and Anticholinesterase Activity in the Rat

The toxicokinetics, tissue distribution, and anticholinesteruse (antiChE ) activity of diazinon were investigated in the rat. Plasma concentrations most adequately fitted a two-compartment open model after iv administration of 10 mg/kg and a one-compartment model after oral administration of 80 mg/kg. Diazinon elimination half-life following iv and oral dosing was 4.70 and 2.86 h, respectively. The oral bioavailabllity was found to be low (35.5%). Hepatic extraction ratios after iv administration of 5 or 10 mg

Long-Term Animal Feeding Trial of the Refined Konjac Meal Ⅱ. Effects of the Refined Konjac Meal on the Aging of the Brain, Liver, and Cardiovascular Tissue Cells in Rats

Rats of both sexes were fed on a basal feed containing 1% refined konjac meal (RKM) for 18 months and the effects of RKM on the cell aging were observed. A comparable group fed on the basic feed was used as the control. Results obtained demonstrate that the long-term feeding of RKM to rats can delay the course of cell aging of the gliocyte, cadiomyocyte, and the endothelial cell of the large and medium arteriases, hence it is likely to delay the occurrence of arteriosclerosis and improve the functions of the brain, heart and vascular system.

Comparative Study of Bovine Pericardium and Gore-Tex in Tissual Interaction with Wistar Rats Diaphragm

Introduction: Simple suture isn’t always possible in large congenital diaphragmatic hernia (cdh) defects. Synthetic materials are used for correction such as Silastic&reg, Gore-Tex&reg (GT), Teflon&reg or biological, such as autologous muscle patches. It was shown that bovine pericardium (bp) was effective to correct those large defects with many positive outcomes when compared with syntactic materials. Aim: This study aims to establish an experimental model of correction for large diaphragmatic defect with PB and GT patches to compare histologically the tissue interaction between them and diaphragm in young Wistar rats. Materials & Methods: 15 wistar rats were divided in 3 groups: Rats that used BP was named G1;Rats that used GT was named G2;and rats with only scraping in the diphragm, named G3 (control). Animals were submited to a laparotomy and fixed pathces to diaphragms and harvested 3 weeks later. Area between normal diaphragm and patches were isolated and separated for histological analysis, such as lymphocytic infiltration (inflammation), neovascularization and fibrosis. Results: G1 presented inflammation between BP and Diaphragm In 5 Samples. G2 Presented Neovascularization In 5 Samples, But No inflammation. Fibrotic tissue overlapping GT patches occurred in 3 samples in G2. Comparing G1 with G2 there was a significant statistical difference concerning inflammation (P = 0.0079), in G1. Comparing neovascularization there is no significant statistical difference (P = 0.4444), despite a slight higher incidence in G2. Fibrosis in both groups presented no significant statistical difference (P = 0.4444), despite a slight higher incidence in G2. There were no alterations in G3. Discussion: Despite the statistical difference in the inflammatory process was more frequent in G1 (P = 0.0079), neovascularization and fibrosis were more frequent in G2. Conclusion: The proposed experimental model was satisfactory to reproduce suture of patches in the diaphragm. These results suggests that inflammation, neovascularization and fibrosis indeed contribute to a benign healing process that reacts differently in each group but can drive to a more lasting and permanent results when biological patch is considered. Statistical report suggests that this study should be continued with a larger sample of animals and a wider period of time before harvest.

Therapeutic Potential of Bone Marrow Derived Mesenchymal Stem Cells in Modulating Astroglyosis of Surgical Induced Experimental Spinal Cord Injury

Background: Spinal cord injury (SCI) unsuccessful regeneration was due to glial scar development. It was a major obstacle to axonal restoration. Safe therapeutic intervention by the use of bone marrow derived stem cells (BMMSCs) transplantation applied in the present study could reduce spinal disability. Material and methods: Forty male albino rats were divided into four groups: GI: negative control (n = 10 rats);GII: positive control after SCI (n = 10 rats);GIII: SCI + BM - MSCs intravenous injected and GIV: SCI + BM - MSCs intra lesion injected (n = 10 rats in each group). The samples were taken from spinal cord tissues around the region of injury and were subjected to histological, immunohistochemical assessment. RNA extraction and real time PCR for detection of nerve regeneration and astrocyte response to the injury were also performed. Results: Clinical improvement occurred by the enhancement in the Basso, Beattie and Bresnahan (BBB) score after SCI. Histological examinations showed positive regenerative responses in GIV compared to GIII. Conclusion: BM-MSCs transplantation has a promising role in enhancing the microenvironment for nerve regeneration through stumbling the glial scaring formation and inflammatory response after chronic spinal cord injury especially by using intra-lesion route injection.

Effect of oxidative stress-associated damage to the lung tissue caused by different body mass index in the rat models

Objective To investigate the influence of different diets on serum protein expression levels of 4-hydroxynonenal(4-HNE),thioredoxin(Trx),thioredoxin reductase(TrxR)and the sctivities of Trx and TrxR,and to explore the effect of damage to the lung tissue and the underlying mechanisms of different body mass index caused by different diets in the rat models.Methods