NS Control Group Research Articles

Memory protective effect of Phyllanthus emblica against scopolamine induced spatial memory loss in male Long Evan Rats tested by Radial Arm Maze

Background: Dementia is the commonest problem of middle aged and elderly subjects due to age related changes and oxidative stress. Many traditional herbal agents including Emblica Officinalis (Phyllantus Emblica) demonstrated its benefit against memory loss. Objective: To investigate the protective effect of Ethanolic extract of Phyllantus Emblica (EEPE) or Emblica Officinalis in scopolamine induced memory impaired male Long Evan rats. Methods: This animal behavioral study was carried out on total 18 male long Evan rats weighing 300-400gm. According to treatment they were divided into NS (control) group (n=6, normal saline 5ml/kg) Sco(n=6; Scopolamine 2mg/kg) and PE(n=6; EEPE 400mg/kg+ Scopolamine 2mg/kg).This experiment was conducted for total 33 days. After 7 days acclimatization NS group and PE rats were treated with normal saline and EEPE respectively for consecutive 26 days. Scopolamine was given to sco group and PE group from day 22 to 26 (5 days during acquisition phase. Spatial memory (working and reference memory error) was assessed by 8 arm radial arm maze (RAM). Statistical analysis was done by ANOVA followed by Bonferroni’s post hoc test. Result: Working memory error (WME) as well as reference memory error (RME) was found significantly higher in scopolamine treated rats compared to normal memory as well as PE pretreated rats whereas no significant difference was found in WME and RME between normal memory rats and PE pre- treated rats. Conclusion: Phyllanthus Emblica can prevent memory impairment in scopolamine induced memory loss in Long Evan rats. J Bangladesh Soc Physiol 2022;17(2): 101-110

GnRH-a-Induced Perimenopausal Rat Modeling and Black Cohosh Preparations' Effect on Rat's Reproductive Endocrine.

GnRH-a-Induced Perimenopausal Rat Modeling and Black Cohosh Preparations' Effect on Rat's Reproductive Endocrine.

PM2.5 inducing myocardial fibrosis mediated by Ang II/ERK1/2/TGF-β1 signaling pathway in mice model.

Objects: To discuss the influence of PM2.5 on myocardial fibrosis and related mechanism. Methods: PM2.5 particles were prepared into different concentrations of solution to drip into the mice’s trachea twice each week. The mice were divided into five groups, Blank control group (C group), NS control group (J group), high dose group (G group, 10 mg/kg), medium dose group (Z group, 5 mg/kg), and 1ow dose group (D group, 2.5 mg/kg). After 6 weeks, the myocardial fibrosis was observed by HE and Masson staining. The expression of Ang II, ERK1/2, and TGF-β1 was examined by Western Blotting (WB) and Real time PCR (RT-PCR). Results: The higher dose PM2.5 was administrated, the worse the myocardial fibrosis was in PM2.5 groups. The expression of Ang II, ERK1/2, and TGF-β1 was increased in higher dose groups in protein and mRNA level. Conclusion: 1. PM2.5 induced the cardiac fibrosis. 2. PM2.5 dripped into trachea in mice model activated the expression of Ang II, ERK1/2, and TGF-β1. The activation of renin-angiotensin system (RAS) was suggested to participate in the cardiac fibrosis induced by PM2.5.

A Comparative Study of Intravenous Clonidine and Dexmedetomidine on Characteristics of Bupivacaine Spinal Anesthesia

Background: Spinal anesthesia with bupivacaine is administered routinely for lower abdominal and lower limb surgeries. The ensuing nerve block is sufficient to ensure patient’s well being, while motor block facilitates the surgeon’s work. In patients receiving spinal anaesthesia, with local anesthetic agents like bupivacaine, the addition of another drug as adjuvant prolongs the analgesia. The present study is designed to study the effect of dexmedetomidine (D) and Clonidine (C) on the duration of motor and sensory block as well as postoperative analgesia by intrathecal bupivacaine in patients undergoing lower limb orthopedic surgery. Subjects and Methods: This study was carried out on 90 patients in the age group of 18 to 50 years, belonging to the American Society of Anesthesiologists(ASA) physical status I and II presenting for lower limb orthopedic surgery were included in the study. Patients were divided randomly into 3 groups. Group D received 1μg kg-1of dexmedetomidine, group C received 2 μg kg-1 of clonidine and group NS Control group received an equivalent amount of normal saline. Results: From the current study we observed that single dose of 1 μg kg-1 dexmedetomidine over 20 minutes started 20 minutes after spinal block and a single dose of 2 μg kg-1 Clonidine resulted in a significant prolongation of time to two-segment regression, postoperative analgesia, sensory block and motor block with the maintenance of hemodynamic parameters. Intravenous dexmedetomidine was more effective than Clonidine at the prolongation of time to two-segment regression, postoperative analgesia, sensory block and motor block of spinal anesthesia with 0.5% hyperbaric bupivacaine. Conclusion: Dexmedetomidine (1μg kg-1) in comparison to Clonidine (2 μg kg-1) and placebo is far more effective in the motor blockade, sensory blockade and duration of postoperative analgesia.

Pyrrolidine dithiocarbamate and dexamethasone are novel treatments of Acute Exogenous Lipoid Pneumonia

BackgroundAcute exogenous lipoid pneumonia (AELP) is characterized by pulmonary inflammation. This mainly occur in children who have ingested sewing machine oil or other mineral oils accidentally. Despite emerging evidences revealing that inhibiting inflammation improves acute exogenous lipoid pneumonia, the actual process of inhibiting inflammation remains unknown. This study aimed to evaluate the effects of PDTC and dexamethasone on AELP to gain insight into the mechanism of AELP. MethodsThe experimental rats were randomly divided into 10 groups: NS control group (NS3 group, NS5 group), Oil inhalation group (AE3 group, AE5 group), PDTC intervention group (PDTC3 group, PDTC5 group), DXM intervention group (DXM3 group, DXM5 group), PDTC + DXM combined intervention group (PDTC + DXM3 group, PDTC + DXM 5 group). Enzyme-linked immunosorbent assay (ELISA) was used to determine concentrations of macrophage migration inhibitory factor (MIF), interleukin-6 (IL-6) and tumor necrosis factor-α (TNFα) in bronchoalveolar lavage fluid (BALF) and serum samples. On the other hand, western blotting was used to measure the expression levels of nuclear factor-κB p65 (NF-κB p65) and b-cell leukemia 2 (Bcl-2) in the lungs. Hematoxylin and Eosin (H&E) staining was performed to evaluate changes in the lung tissue. The wet-to-dry lung weight ratio was subsequently used to determine the pulmonary edema of the lungs. ResultsThere were increased MIF levels in both serum and BALF samples of the AE group. Pyrrolidine dithiocarbamate (PDTC) and dexamethasone (DXM) independently and in combination reduced pulmonary inflammation induced by the sewing machine oil by regulating MIF expression. TNF-α and IL-6 levels in serum and BALF samples of the AE group were higher than those of the NS control animals. However, their levels decreased after treatment with either PDTC, DXM or PDTC + DXM. Similarly, NF-κBp65 expression increased after oil inhalation but decreased after treatment with either PDTC, DXM or PDTC + DXM. PDTC, DXM and PDTC + DXM treatment significantly reduced pulmonary inflammation and pulmonary edema of the lung tissue following induction of acute exogenous lipoid pneumonia. ConclusionsIndividual or combined use of PDTC and DXM can ameliorate pulmonary inflammation induced by inhalation of sewing machine oil by inhibiting the NF-κB pathway in young rats. These findings provide novel insights that will greatly contribute in treatment of AELP.

The effects of CTX damage or inhibition of bone marrow hematopoiesis and GM-CSF stimulation of bone marrow hematopoiesis on the peripheral blood TCRβ CDR3 repertoire of BALB/c mice

Objective: This paper aims to investigate the dynamic changes of the T-cell receptor (TCR) β complementarity-determining region 3 (CDR3) repertoire during cyclophosphamide or Cytoxan (CTX) damage or inhibition of bone marrow hematopoiesis caused by a reduction of peripheral blood white blood cells (WBCs) in BALB/c mice.Methods: We analyze TCR CDR3 repertoire of BALB/c mice including (1) NS control group (2) CTX damage group (3) CTX damage + GM-CSF recovery group (4) CTX damage + auto-recovery group.Results: The number of WBCs in the CTX group is significantly lower than that in the NS group and after GM-CSF injection, the GM-CSF group is higher than that in the NS group. The diversity of the CTX damage group is the highest and there is a significant difference in high-frequency clonal proliferation between the CTX damage group and CTX damage + GM-CSF recovery group compared with the NS control group. In addition, the numbers of unique productive CDR3 overlapping numbers in the four experimental groups are similar.Conclusions: These data reveal that CTX significantly reduced the number of WBCs and ratio of high-frequency TCR CDR3 sequences, and indirectly increased the diversity of the TCR CDR3 repertoire. GM-CSF quickly restored the number of WBCs, and partially restored changes in the TCR CDR3 repertoire induced by CTX. Results from monitoring the dynamic changes of the TCR CDR3 repertoire can be used to assess the effects of CTX and GM-CSF on the function of peripheral blood T cells and to explore the possible underlying mechanisms.

Effects of prenatal cold stress on maternal serum metabolomics in rats

Our previous studies have shown that prenatal cold stress leads to placental inflammatory response and induces anxiety-like behavior reduced in offspring rats. However, the role and mechanisms by which prenatal cold stress affects offspring remain unclear. The aim of this study was to determine the metabolic profiles from the maternal serum and helpful in understanding the role and mechanisms by which prenatal cold stress affects the offspring. In this study, liquid chromatography-mass spectrometry (LC/MS) was used to analyze serum metabolites, and PCA, PLS-DA, and OPLS-DA were performed to analyze changes in metabolites in the maternal serum after cold stress of 3 or 7 days. The results showed that 19 metabolites in the CS (cold stress 7 days)-NS (control) group and 23 metabolites in the CT (cold stress 3 days)-NT (control) group were significantly altered. These metabolites were mainly associated with unsaturated fatty acid synthesis, and arachidonic acid, linoleic acid, and glutamine and glutamate metabolism. The data indicated that prenatal cold stress not only affected the maternal neuroendocrine system, but also affected the immune system, and lipid and amino acid metabolism. These results further supported the findings of our previous studies on the effects of prenatal cold stress on the mother and offspring. A more comprehensive understanding of these data may lead to maternal intervention that can reverse the damage of prenatal stressors.

Construction and Evaluation of a Tuberculosis Multistage Vaccine Candidate Based on Recombinant Listeria ivanovii

To construct a recombinant Listeria ivanovii (LI) strain that expressed Mycobacterium tuberculosis (MTB) specific antigen protein as a novel multistage tuberculosis (TB) vaccine candidate, and evaluate the biosafety and immunogenicity in mouse model. T cell epitopes of four genes related to different stages of MTB infection were fused in series to form an antigen gene, i.e. the multistage antigen gene (named msv). Then msv was inserted into the targeting plasmid that contained LI homologous sequences. Recombinant LI strain was obtained by transfecting LI with targeting plasmid and screening the recombinant LI strain that carried msv in the genome after series of homologous gene recombination processes. The growth rate of the recombinant LI strain in vitro was observed and the expression of target protein was verified by Western blot. The 50% lethal dose (LD 50) of the recombinant strain to C57BL/6 mice was measured. Mice were intravenously inoculated with vaccine candidate in dose of 0.1×LD 50.The serum alanine aminotransferase (ALT) levels, bacterial load in organs, and organ pathological sections before and 1, 2, 3, 5, 7, 14 d after vaccination were used to evaluate the safety of vaccine candidate strain. To analyze the immunogenicity of vaccine candidate strain, mice were intravenously inoculated with LI- msv, LI, and NS respectively. Nine days post immunization, the spleens were isolated under sterile conditions and splenocytes were collected and stimulated. Lyphocytes which secret specific cytokines, interferon (IFN)-γ, tumor necrosis factor (TNF)-α and interleukin (IL)-2 were analyzed by flow cytometry. A recombinant strain named LI- msv which was capable of expressing the multistage TB antigen protein was successfully constructed. The LD 50 value of LI- msv for C57BL/6 mice (i.v.) was 3.3×10 8 CFU. After intravenously immunized the mice, this strain mainly multiplied in the liver and spleen, and was cleared at 7 d post innoculation. Such infection process caused transient pathological damages of the liver and spleen. Results of flow cytometry showed specific IFN-γ + CD4 + and IFN-γ + CD8 +T lymphocytes were successfully induced in LI -msv immunized mice spleen lymphocytes. The frequency of IFN-γ positive CD4 + and CD8 +T cells was significantly higher than those of vector control group and NS control group ( P<0.005). Additionally, the frequency of specific TNF-α + CD4 + T cell in LI -msv immunized group was significantly higher than that of vector control ( P<0.01) and NS control group ( P<0.005), and TNF-α + CD8 + T cell frequency obviously increased than NS control group ( P<0.005). A novel multistage TB vaccine candidate expressing TB multistage antigen based on LI was successfully constructed. This vaccine candidate is safe and can induce specific cellular immune response to some extent. It is promising to be further studied as a candidate vaccine against tuberculosis.

Antisense oligonucleotide against insulin-like growth factor I receptor inhibits tumor growth in human hepatocellular carcinoma orthotopic implantation models and its influence on the blood indicators in nude mice in vivo

Objective To investigate the inhibitory effect of antisense oligonucleotide against insulin-like growth factor 1 receptor (IGR-1R) on tumor growth in human hepatocellular carcinoma orthotopic implantation models and its influence on the blood indicators in nude mice. Methods We did the experiment 3 times, and 40 models for each time were divided into 5 groups randomly: NS control group, three IGF-1R antisense oligdeoxynucleotides (ASODN) [75, 50, 25 mg/(kg·day)] groups and 5-fluorouracil (5-Fu) control group. Eighty nude mice served as normal control group. The agents were given for 25 times through intravenous injection in human hepatocellular carcinoma orthotopic implantation models. When the experiment was over, the volume and weight of tumor were measure, the inhibitory rate of tumor growth was calculate, and the pathomorphological change was examined. Alpha fetal protein (AFP), routine blood test and biochemical indexes were determined in three experiments respectively. Results The average volume and weight of tumor in IGF-1R ASODN group was significantly reduced as compared with those in NS control group. The inhibitory rate of tumor growth in 75 mg/(kg·day) IGF-1R APSODN group in three experiments were 76.36%, 71.81% and 78.81%, 55.65%, 61.74% and 63.58% in 50 mg/(kg·day) group, and 47.72%, 50.34% and 54.30% in 25 mg/(kg·day) group respectively. Large piece of necrosis and fibrosis could be seen under a light microscope in IGF-1R ASODN groups. AFP in IGF-1R ASODN groups was lower (P<0.05) than in NS control group, PLT in IGF-1R ASODN group [75 mg/(kg·day)] was lower than in NS control group (P<0.05), and GLU in IGF-1R ASODN group [75 mg/(kg·day) and 50 mg/(kg·day)] was higher than in normal control group (P<0.05). There was no significant difference between IGF-1R ASODN group and normal control group in other blood indicators. Conclusion IGF-1R ASODN is an anti-hepatoma biological agent with high efficiency and low side-effect. Key words: Carcinoma, hepatocellular; Insulin-like growth factor 1 receptor; Antisense oligdeoxynucleotides; Inhibitory rates of tumor growth

Dexmedetomidine attenuates the propofol-induced long-term neurotoxicity in the developing brain of rats by enhancing the PI3K/Akt signaling pathway.

BackgroundPropofol induces short- and long-term neurotoxicity. Our previous study showed that dexmedetomidine (Dex) can attenuate the propofol-induced acute neurotoxicity in rodents by enhancing the PI3K/Akt signaling. However, whether treatment of young rats with Dex could protect them from long-term neurotoxicity induced by propofol is unclear.Materials and methodsSeven-day-old male Sprague Dawley rats were randomized and injected intraperitoneally with saline (100 μL, NS), propofol (100 mg/kg), Dex (75 μg/kg), propofol (100 mg/kg) plus Dex (25, 50 or 75 μg/kg), 10% dimethyl sulfoxide (DMSO, 100 μL) or TDZD-8 (a GSK3β inhibitor, 1 mg/kg), or intracerebroventricularly with DMSO (5 μL) or LY294002 (a PI3K inhibitor, 25 μg/5 μL DMSO). Other rats in the experimental group were injected with the same doses of propofol, Dex and LY294002 or TDZD-8. All the rats were monitored until they were 9 weeks old. Their spatial learning and memory were tested by Morris water maze. The neuronal apoptosis, expression of PSD95, expression and phosphorylation of Akt and GSK3β and synaptic ultrastructures were determined by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling, immunohistochemistry, Western blot and transmission electron microscopy assays, respectively.ResultsCompared with the NS control group, young rats injected with intralipid, Dex, TDZD-8, LY294002 or DMSO alone did not show any significant change as they aged. Propofol significantly increased the escape latency time, hippocampal neuroapoptosis and synaptic ultrastructural changes but decreased the relative levels of PSD95 expression, and Akt and GSK3β phosphorylation in the developing hippocampus of the rats. The neuronal toxic effects of propofol were significantly mitigated by the pretreatment with a higher dose of Dex. The neuroprotective effect of Dex was enhanced by the treatment with TDZD-8, but was completely abrogated by the treatment with LY294002.ConclusionOur results indicated that the pretreatment of young rats with Dex attenuated the propofol-induced long-term neurotoxicity in their developing hippocampus by enhancing the PI3K/Akt signaling.

High Salt Diet May Stimulate Fructose Uptake in Brain Neurons and Contribute to Neuronal Apoptosis

Clinical studies have been leaning toward the role that the central nervous system (CNS) is one of the main controls of blood pressure regulation such as the onset of fructose‐induced hypertension. The CNS is by far the most energy demanding and consuming organ in our body and it can metabolize fructose but that has consequences. However, the connection between fructose and hypertension within the CNS is still lacking in definition. Here we tested the hypothesis that a high fructose diet contributes to hypertension via increase neuronal apoptosis. Eight‐week‐old male Sprague Dawley (SD) rats were divided into 4 groups and were fed a normal salt (NS: 0.4% NaCI chow), high salt (HS: 4% NaCl chow), NS diet with fructose (F: 20% fructose drinking water), and high salt plus fructose (HS+F: 4%NaCl chow and 20% drinking water) diet, respectively, for 3 consecutive weeks. Their blood pressure was measured prior and post each diet treatment using the non‐invasive tail‐cuff technologies. All rats were then euthanized, then their blood and cerebrospinal fluid (CSF) was withdrawn for fructose measurements. HS+F rats showed an increase in mean arterial blood pressure (MAP) 126 mmHg compared to control 90 mmHg (p&lt;0.0001). There is no significant MAP alterations in HS or F alone groups compared to NS group. Fructose measurement demonstrated that the serum fructose concentration is significantly increased in the F group (359 μM), and HS+F (409 μM) compared to control NS group (253 μM), while no significant alterations were observed in serum fructose in HS alone group (229 μM). In contrast, CSF fructose concentration is dramatically decreased in F (230 μM) and HS+F (231 μM) groups compared to NS control group (290 μM). In addition, immunostaining showed that HS+F group significantly promoted immunoreactivity of GLUT5, a fructose transporter in the brain paraventricular nucleus (PVN). Furthermore, incubation of brain neurons isolated from neonatal SD rats with 1 mM fructose for 6 hours stimulated neuronal apoptosis (27%) compared to the control (20%, P&lt;0.05). In summary, this study demonstrated that HS+F diet decreases CSF fructose concentration, increases PVN GLUT5 expression, and results in hypertension in normal SD rats. These results suggest that HS facilitates brain neurons to uptake fructose into neuronal cells, resulting in an increase in intracellular fructose, which in turn, stimulate cell apoptosis. Increased neuronal apoptosis may contribute to hypertension.Support or Funding InformationFunding support: R15‐HL122952, Qinghui Chen; R15‐HL129213, Zhiying Shan; Michigan Tech and portage health Foundation REF, Zhiying ShanThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Effect of Corydalis Rhizoma and L-tetrahydropalmatine on dopamine system of hippocampus and striatum in morphine-induced conditioned place preference rats

To investigate the effects of Corydalis Rhizoma and L-tetrahydropalma-tine (L-THP) on the levels of dopamine neurotransmitter (DA), dopamine transporter (DAT) and the second dopamine receptor (D2R) in learning and memory-related brain areas, hippocampus and striatum, the DA, DAT and D2R were detected in conditioned place preference (CPP) rats suffered from morphine. And comparation the degree of similarity and consistency of the pharmacological effects was also studied. The rats were trained in black compartments and white ones (drug-paired compartment) with the increasing doses of morphine for 10 days (hypodermically injected from 10 mg•kg⁻¹ to 100 mg•kg⁻¹). Models of CPP were validated in those psychological dependence rats after 48 h training. The dopamine contents were detected as soon as the materials of hippocampus and striatum are harvested from rats of NS control group and model group. The DAT and D2R levels are measured by Western blot. The high, medium and low dose group of Corydalis Rhizoma are given Corydalis Rhizoma 2, 1, 0.5 g•kg⁻¹ water extraction liquid respectively (which contains L-THP were 0.274, 0.137 and 0.137 mg respectively), and the high, medium and low dose group of L-THP were given L-THP 3.76, 1.88, 0.94 mg•kg⁻¹ lavage treatment respectively, NS treatment group were lavaged normal saline for 6 days and they were killed after test of CPP, again tested DA levels and expression of DAT and D2R similar to the front of materials. The reduction effects of CPP were observed in the groups of both Corydalis Rhizoma (2, 1 g•kg⁻¹) and L-THP (3.76, 1.88 mg•kg⁻¹) subjected to medicine for 6 days (P<0.01). Compared with the NS treatment group and the model group, the higher values including in the contents of neurotransmitter dopamine were detected of hippocampus and striatum (P<0.01, P<0.05), the DAT and D2R protein expression of Corydalis Rhizoma (2, 1 g•kg⁻¹) and L-THP (3.76, 1.88 mg•kg⁻¹) increased in hippocampus and striatum (P<0.01). Learning and memory-related brain regions hippocampus and striatum was another neuroanatomical sites of action in the treatment of mental dependence of fumarate and L-THP, its mechanism was related to lowering its elevated DA neurotransmitter levels, and increasing the expression of DAT and D2R. Corydalis Rhizoma could be play 14-times roles in effect of L-THP. The similar effects were observed on the neurotransmitter dopamine, DAT and D2R in learning and memory-related brain areas, hippocampus and striatum of the morphine- dependent rats.

Role of α7 nicotinic acetylcholine receptor in attenuation of endotoxin induced delirium with dexmedetomidine in mice

To observe the role of α7 nicotinic acetylcholine receptor (α7nAChR) in the protection against delirium by the use of dexmedetomidine (DEX) in endotoxin derived delirium and its mechanism. 100 male adult C57BL/6 mice were randomly divided into normal saline control group (NS group), DEX control group, lipopolysaccharide (LPS) induced endotoxemia model group (LPS group), DEX protection group (DEX+LPS group), and α-bungarotoxin antagonism group (α-BGT+DEX+LPS group), with 20 mice in each group. A model of endotoxemia was reproduced by intraperitoneal injection of LPS 20 mg/kg, and the mice in NS group and DEX control group were given equivalent sterile normal saline. The mice in DEX control group, DEX+LPS group, and α-BGT+DEX+LPS group were intraperitoneally injected with DEX 40 μg/kg 15 minutes before LPS injection. The mice in α-BGT+DEX+LPS group were intraperitoneally injected with α7nAChR inhibitor α-BGT 1 μg/kg 15 minutes before DEX injection. The mice in NS group were given equivalent sterile normal saline. Ten mice in each group were assigned for open field test before and 24 hours after model reproduction, and the mice were then sacrificed to obtain the specimens. The levels of tumor necrosis factor-α (TNF-α) and neuron-specific enolase (NSE) in serum were determined by enzyme-linked immune sorbent assay (ELISA). Western Blot method was used to determine the expression of choline acetyltransferase (ChAT) and acetylcholinesterase (AChE) in hippocampus. Another 10 mice were subjected to new object recognition test to observe the total exploration time during training period and preference index at 3 hours and 24 hours after LPS challenge. There were no significant differences in all parameters between NS group and DEX control group. (1) It was shown by the open field test results that there were no significant differences in all parameters of open field test before model reproduction among all the groups. Twenty-four hours after model reproduction, when compared with NS group, the mice in LPS group showed that they had the ability of cognition of new environment, but learning and memory abilities were lowered, and tension was increased. DEX could significantly attenuate the degree of delirium, however, the protection of DEX from the delirious syndrome was antagonized partly by α-BGT. (2) The new object recognition test results showed that compared with NS group, the ability of exploring new object was decreased in LPS group. DEX could significantly improve the exploration ability. However, DEX failed to control the delirious syndrome in α-BGT+DEX+LPS group. (3) The results of ELISA showed that the levels of TNF-α and NSE in serum were significantly increased in LPS groups as compared with that in NS group, and the levels of TNF-α and NSE were significantly decreased in DEX+LPS group. However, α-BGT could antagonise the protective effect of DEX [TNF-α (ng/L) in NS, LPS, DEX+LPS and α-BGT+DEX+LPS groups was 23.72±3.13, 808.78±87.86, 192.96±31.47, 829.99±80.98, respectively, and NSE (μg/L) was 8.70±0.74, 25.90±3.03, 18.10±2.14, and 23.12±2.21, respectively, all P < 0.01]. (4) The results of Western Blot showed that compared with NS group, the protein expression of ChAT in LPS group was significantly declined, and the protein expression of AChE was significantly increased. DEX could reverse the expressions of ChAT and AChT, however, α-BGT could reverse the protective effect of DEX [ChAT (gray value) in NS, LPS, DEX+LPS and α-BGT+DEX+LPS groups was 1.536±0.150, 0.381±0.138, 0.914±0.173, 0.628±0.088, respectively, and AChE (gray value) was 0.382±0.201, 1.843±0.325, 0.898±0.155, and 1.470±0.220, respectively, P < 0.05 or P<0.01]. Delirium syndrome may occur in mice with endotoxemia. DEX could attenuate endotoxemia-associated delirium syndrome through transforming central neurotransmitter, and its mechanism maybe related with α7nAChR.

Effects of rotigaptide (ZP123) on connexin43 remodeling in canine ventricular fibrillation.

The present study investigated the effects of rotigaptide (ZP123) on the expression, distribution and phosphorylation of connexin43 (Cx43) in myocardial cell membranes in cardioversion of ventricular fibrillation (VF). A model of prolonged VF (8, 12 and 30 min) was established in mongrel dogs (n=8/group), following treatment with ZP123 or normal saline (NS control). A sham control was included. Cardiopulmonary resuscitation was begun at the start of VF followed by defibrillation. Animals received a maximum of three defibrillations of increasing energy (70, 100 and 150 J biphasic shock) as required. The average defibrillation energy, defibrillation success rate, return of spontaneous circulation and survival rate were recorded. Cx43 and phosphorylated (p-) Cx43 expression in cardiomyocyte membranes was detected by western blot and immunofluorescence analyses. Compared with the NS-treated control groups, the success defibrillation rate in the 8-min and 12-min ZP123 groups was significantly higher (P<0.05), while the average defibrillation energy was significantly lower (P<0.05). Cx43 expression in the VF groups was significantly lower than that in the sham control group (P<0.05). Cx43 expression was higher in the 12-min and 30-min ZP123 groups than that in the NS control group (P<0.05), while p-Cx43 expression decreased, although the levels were significantly higher than those in the control groups (P<0.05). Cx43 expression was positively correlated with the defibrillation success rate (r=0.91; P<0.01) and negatively with the mean defibrillation energy (r=−0.854; P<0.01), while p-Cx43 expression was positively correlated with the success rate of the previous three defibrillations (r=0.926; P<0.01). In conclusion, ZP123 reduced Cx43 remodeling through regulating the expression, distribution and phosphorylation of Cx43, thereby reducing the defibrillation energy required for successful cardioversion.

Trigeminal Neuralgia Induced by Cobra Venom in the Rat Leads to Deficits in Abilities of Spatial Learning and Memory

Background: Patients with chronic pain usually suffer from cognitive impairment, with memory deterioration being the most common deficit that affects daily functioning and quality of life. The causes for this impairment are not clear despite intensive clinical studies. Few studies have evaluated impaired learning using animal models of persistent pain. Objective: In this study, a new trigeminal neuralgia model induced by cobra venom was adopted to explore effects of chronic pain on spatial learning and memory in rats. Study Design: Controlled animal study. Setting: Department of Anesthesiology, Pain Medicine &amp; Critical Care Medicine, Aviation General Hospital of China Medical University. Methods: Thirty adult male Sprague-Dawley rats were randomly divided into 2 groups (n = 15): NS control group and cobra venom group, 0.9% sterile saline or cobra venom solution was injected into the sheath of the infraorbital nerve (ION), respectively. The development of trigeminal neuralgia was accessed by changes in free behavioral activity 3 days before the surgery and 3, 7, 12, 20, and 30 days after the surgery to identify whether the model was successful or not. Morris water maze test determined the abilities of spatial learning and memory at the time points before the surgery, and 2 weeks and 5 weeks after the surgery. We also observed the ultrastructure of the ION and medulla oblongata of rats following 8 weeks of chronic trigeminal neuropathic pain. Results: Rats with the cobra venom injection displayed significantly more face grooming and fewer exploratory activities compared to the NS control group or baseline (P &lt; 0.01). Both groups improved their latency to reach the platform with the largest difference on the first day (P &lt; 0.01), but without memory deficits in a probe trial for the second water maze protocol. For the third water maze testing, the rats in the cobra venom group experienced decreased abilities of spatial learning and memory, a longer latency with spatial memory deficits during the probe trial (P &lt; 0.05). At the ultrastructural level, we found changes in the medulla oblongata after cobra venom injection resulting in severe demyelination and loss of axons that might be implicated in the causes of cognitive deficits. Limitations: Limitations include partial vision loss in the eye on the lesion side of the rats that might be missed and the absence of evaluating the ultrastructural changes in other parts of the brain. Conclusions: The results of this study suggest that trigeminal neuralgia induced by cobra venom in adult rats can impair spatial learning and memory function over time and results in demonstrable changes in the ultrastructure of the medulla oblongata. This new animal model may be useful for future studies on the effect of chronic pain on learning and cognition. Key words: Cognitive deficits, memory deterioration, cobra venom, trigeminal neuralgia, electron microscopy

Effects of musk ketone on nerve recovery after spinal cord injury.

The present study aimed to determine the effects of musk ketone on nerve recovery in rats after spinal cord injury. A total of 105 SD female rats were used to establish the rat with dorsal spinal cord injury model (modified Allen's method). The rats weighed from 200 to 250 g and were provided by the Experimental Animal Center of Chongqing Medical University. They were randomly divided into five treatment groups: saline (NS group), methylprednisolone (MP group), and musk ketone groups (MO1, MO2, and MO3 groups). The Swash plate test and BBB behavioral score were used to determine neurological function recovery after spinal cord injury. Hematoxylin-eosin (HE) staining was used to detect general structural changes in spinal cord tissue. The enzyme-linked immunosorbent assay was used for the determination of interleukin 10 (IL-10) in spinal cord tissue. We found that compared with the NS control group, critical angle, BBB score and IL-10 levels in rat spinal cord tissue significantly increased in the MP group and MO groups 7 and 14 days after the operation. HE staining showed that in the NS group, there was hemorrhage, edema, necrosis, axonal demyelination, inflammatory cell infiltration and glial cell response in spinal cord tissue. After 7 days, spinal cord edema and inflammation were reduced and neuronal degeneration and necrosis were not evident in the MP and MO groups. We conclude that musk ketone can reduce secondary damage after spinal cord injury and promote nerve recovery in rats.

Processing collocations in a second language: A case of first language activation?

ABSTRACTThis study investigated the possible influence of first language (L1) collocational patterns on second language (L2) collocational processing. A lexical decision task was used to assess whether collocational patterns acceptable in the L1 but not the L2 would still be activated when processing language entirely in the L2. The results revealed no such activation. Furthermore, L2 speakers did not produce accelerated processing for control collocations that were acceptable in the L2 but not the L1. Based on these findings, we put forth some theoretical suggestions regarding recent research indicating accelerated processing for congruent over incongruent collocations. Finally, our NS control group revealed some unexpected tendencies that cannot be easily accounted for with our current understanding of L1 language processing.

A study on sexual development of SD rats by using KiSS1RNA interference mediated bylentivirus-based vectors

To explore the possible mechanism of KiSS1 in control GnRH secretion participate in sexual development onset and normal reproduction regulation by investigate the changes of expression of KiSS1, GnRH in hypothalamus and LH, FSH, E2 in serum,by using RNA interference Mediated with Lentivirus-based Vectors, after Interfering expression of KiSS1. Constructed the eukaryotic expression plasmid of rat KiSS1 gene and four plasmid expressing KiSS1 microRNA, then respectively co-transfected them into 293T cells. Real-time PCR detected the expression of KiSS1 mRMA in order to filter the most effective microRNA plasmid.Constructed recombinant entivirus and determined the titer, then they were intracerebroventricularly infused into the brain of Sprague-Dawley rats (21-day-old) . The three groups included interference virus group, Lentivirus-control,NS-control, Ten rats in each group animals were sacrificed at 30-day-old, 35-day-old, 45-day-old. Then the expression of KiSS1 and GnRH mRNA were conducted in the rat hypothalamus with Real-time PCR, the LH, FSH, E2 in serum were examined with Chemiluminescence method. HE staining was used to observe histomorphology ofovarian. Successfully filtered the most effective microRNA plasmid and constructed recombinant lentivirus. The titer of recombinant lentivirus was 8×108TU/ml. The level of KiSS1 mRNA in interference virus group was significantly reduced after infecting recombinant lentivirus compared with control group at 30d NS-control group0.2±0.02, Lentivirus-control group 0.188±0.023, interference virus group 0.106±0.018; at35d NS-control group 0.433±0.046; Lentivirus-control group 0.41±0.034; interference virus group 0.218±0.025; at 45d NS-control group 0.315±0.048; Lentivirus-control group 0.282±0.052; interference virus group 0.215±0.033, at 30d F=112.40 P<0.01; at 35d, F=209.5 P<0.01; at 45d, F=5.2, P<0.05. The level of GnRH mRNA in interference virus group was significantly reduced after infecting recombinant lentivirus compared with control group (at 30d NS-control group 0.387±0.044; Lentivirus-control group 0.373±0.05; interference virus group 0.23±0.03; at35d NS-control group 0.517±0.048; Lentivirus-control group 0.53±0.052; interference virus group 0.407±0.03; at 45d NS-control group 0.468±0.03, Lentivirus-control group 0.479±0.038; interference virus group 0.455±0.054,at 30d,F=24.6, P<0.01at35d,F=20.90 P<0.01). The difference was statistical significance. The level of LH in interference virus group is lower than other two groups at 35d (NS-control group 0.111±0.008 mU/ml ; Lentivirus-control group 0.106±0.006mU/ml; interference virus group 0.101±0.004mU/m F=5.98 P<0.05). The level of FSH in interference virus group is lower than other two groups at 35d (NS-control group 0.219±0.015mU/ml; Lentivirus-control group 0.215±0.014mU/ml; interference virus group 0.205±0.014mU/ml F=9.72 P<0.05). The level of E2 in interference virus group is lower than other two groups(at 35d, NS-control group 56.04±4pg/ml; Lentivirus-control group 52.9±3.26pg/ml; interference virus group 46.8±3.01pg/ml; at45d NS-control group 57.4±5.5pg/ml; Lentivirus-control group 58.1±3.02pg/ml; interference virus group 52.4±3.57pg/ml; at 35d,F=25.25 P<0.01; at 45d,F=7.63 P<0.05). Meanwhile, the time of vaginal orifice opening of the interference virus group was significantly later than control virus group and normal saline group (the interference virus group37.4±1.57d, control virus group 35.2±1.19d, and normal saline group 34.9±0.99d, F=18.1 P<0.01). The histology of ovary showed similar results. Our result show that the lentiviral vector of KiSS1-microRNA can inhibited the expression of KiSS1 stabily, efficiently and specificly. Lentiviral with KiSS1-microRNA can affect the expression of GnRH and The levels of sex hormone. Intralateroventricular microinjection of KiSS1-microRNA Lentiviral can delay sexual development of SD female rat. Lentivirus-mediated RNAi can effectively suppress the expression of KiSS1 stabily,as a result the expression of GnRH gene can be suppressed, then affecting sexual development. It may provide a potential tool for the study of targeting control of sexual development in vivo and treating precocious puberty and other diseases.

Synthesis, characterization, and antitumor evaluation of the albumin–SN38 conjugate

7-Ethyl-10-hydroxycamptothecin (SN38), the active metabolite of irinotecan, exerts a 100-fold to 1000-fold higher effect than irinotecan itself against several tumor cell lines. However, the water insolubility of SN38 has prevented its direct use as an antitumor drug in the clinic. To improve the water solubility and antitumor efficacy, SN38 was covalently attached to the only free sulfhydryl at cysteine-34 on the BSA site specifically through a thiol-binding linker to form a prodrug BSA-SN38 conjugate (BSA : SN38=1 : 1). The water solubility of this conjugate was similar to albumin using the current method. Also, SN38 loading in this conjugate became controllable. Size-exclusion chromatography purification and UV characterization of the SDS-PAGE electrophoresis product were carried out. Then, an MTT assay was carried out to test the antitumor effect of this conjugate on five colon cancer cell lines in vitro. The 72 h IC50 values of the BSA-SN38 conjugate ranged from 1.5 to 6.1 μmol/l. A colorectal peritoneal carcinomatosis model in mice was established to determine the intraperitoneal chemotherapy effect of the BSA-SN38 conjugate. The BSA-SN38 conjugate at an SN38 equivalent dose of 10 mg/kg/day was administrated every 4 days. Eighteen days after manipulation, the mice were euthanized and the tumors in the abdominal cavity were collected and weighed. Tumors in the BSA-SN38 conjugate treatment group (m=0.21 ± 0.15 g) were found to be significantly (P=5) lighter than those in the NS control group (m=4.74±0.73 g). The results indicated that this water-soluble BSA-SN38 conjugate exerted a strong antitumor effect on colorectal carcinoma.

Gabapentin reduces mechanical allodynia in a rat model of tibial bone cancer pain

Objective To explore the effects of gabapentin on mechanical allodynia in rats with tibial bone cancer pain(BCP). Methods Forty-two female SD rats were randomized into 7 groups (n=6): naive group (group N), sham operation+ NS control group (group SN), sham operation+ GBP200mg/(kg·d) group (group SG200), BCP+ NS control group (group BN), BCP+ GBP50mg/(kg·d) group (group BG50), BCP+ GBP100mg/(kg·d) group (group BG100), and BCP+ GBP200mg/(kg·d) group (group BG200). The rats in group N, SN and BN received 5 ml normal saline and the rats in group SG200, BG50, BG100 and BG200 received 200, 50, 100 and 200 mg/(kg·d) dose of GBP via feeding from day 7 to 13 after operation, respectively. Mechanical withdrawal threshold(MWT) of the right paw and behavioral assays for ambulatory pain were measured just before operation and on days 1, 3, 5, 7, 8, 10, 12 and 14 after operation. Results MWT((3.78±0.38)g) in rats with BCP decreased and behavioral assays for ambulatory pain(0.76±0.44) increased on day 7 after operation, as compared with those in group N((14.50±1.38)g, (0.00±0.00))and group sham((10.21±0.88)g, (0.00±0.00))(P 0.05). Compared with those in group BN, there was no change on MWT in group BG50(P>0.05), and however, behavioral assays for ambulatory pain decreased (P<0.05). MWT in group BG100((5.35±0.85)g) and BG200((5.71±0.72)g) increased in day 10 after operation, as compared with those in group BN((2.61±0.40)g) and group BG50((3.28±1.15)g)(P<0.05), and the difference was still statistically significant until day 14 (P<0.05). Behavioral assays for ambulatory pain in group BG100 and BG200 decreased from day 8 after operation, as compared with those in group BN and group BG50(P<0.05). Conclusion Gabapentin, in medium to large dosage, can inhibit pain reaction of rats with bone cancer pain. Nevertheless, with the development of cancer, the effect of gabapentin decreases. Key words: Gabapentin; Bone cancer pain; Mechanical withdrawal threshold; Behavioral assays for ambulatory pain