Objective:To investigate the effects of β-sheet breaker peptide H102 on NF-κB signal pathway in brain of APP/PS1 double transgenic mice. Methods:Thirty 8-week-old APP/PS1 double transgenic mice were randomly divided into model group and treatment group. A group of C57BL/6J mice with the same age and background were served as controls (n=15). H102 5 μl(5.8 mg/kg) was infused by intranasal administration to mice in H102 treatment group, and equal volume of blank solution of H102 (chitosan, BSA) was given to mice in control group and model group. After 16 weeks, the ability of spatial reference memory was tested by Morris Water Maze. Then immunohistochemistry tests and Western blot technique were used to detect the content of amyloid beta peptide 1-42(Aβ_1-42), nuclear factor-kappa B (NF-κB), inhibitor of NF-κB (IκB), IκB kinase (IKK), the corresponding phosphorylated proteins (p-NF-κB、p-IκB、p-IKK), inducible nitric oxide synthase (iNOS) and cleaved Caspase 3 proteins in mice brain. Results:①The ability of learning and memory was significantly lowered in model group than that in control group. And the ability of learning and memory was significantly improved in treatment group than that in model group (P<0.05). ②The contents of Aβ_1-42, p-IKK, p-NF-κB, p-IκB, intranuclear NF-κB,iNOS and cleaved Caspase 3 in mouse brain were significantly increased in model group than those of control group, and these protein expressions were significantly lowered in treatment group than those in model group (P<0.05). Conclusion:H102 can inhibit NF-κB signal pathway in brain of APP/PS1 double transgenic mice, reduce the levels of inflammation and apoptosis in nerve cells, and improve the ability of learning and memory in transgenic AD mice.

Objective:To observe the effect of AdipoRon for the treatment of type 2 diabetes (T2DM)in mice and its effect on the liver. Methods:Forty male C57/BL6 mice (SPF) were randomly divided to normal control (NC) group and the experimental group. To establish the T2DM mice model, mice in the experimental group were fed with high fat and high glucose, combined with intraperitoneal injection of streptozotocin (STZ) in small doses, and mice were further subdivided into model control (DM) group, model control with low AdipoRon (DM+L) group and model control with high AdipoRon (DM+H) group (n=10). Serum indexes, such as levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP) were detected biochemically and the morphological changes of liver cells were observed with HE staining and expression of liver carbohydrate related gene (PEPCK) were determined by real-time fluorescence quantitative PCR (real time FQ-PCR). Results:Compared with mice in the DM group, levels of ALT, AST, ALP, triglyceride (TG), glucose (GLU) reduced in DM+L and DM+H group (P<0.05). Concentrations of serum free fatty acids (FFA) in DM+L and DM+H group reduced significantly (P<0.05). Besides, concentrations of liver glucose-6-phosphatase (G-6-P) in the mice of DM+L group reduced significantly, while there was no significant difference in the content of G-6-P between the mice of DM+H group and the mice of DM group. Furthermore, the expression of the liver phosphoenolpyruvate carboxylase (PEPCK) in the DM+H group reduced significantly (P<0.05). Compared with the DM group no significant change was found in the PEPCK expression between DM+L and DM group. Conclusion:The serum indexes such as levels of ALT, AST, ALP, TG, Glu, G-6-P and PEPCK were all reduced in DM mice treated with AdipoRon, indicating the obvious protecting effect of AdipoRon on the liver in DM mice.

Objective:To investigate the effect of rosuvastatin therapy on C-C chemokine receptor(CCR2)expression in mononuclear cells in patients with carotid atherosclerosis and explore the possible upstream mechanism. Methods:Twenty patients without previous statin treatment were enrolled. Rosuvastatin were given 5 to 20 mg/day for 3 months. At baseline and 12 weeks, lipid profile and plasma monocyte chemotactic protein-1 (MCP-1) levels were examined. The mRNA and protein expressions of CCR2 in the mononuclear cells were measured with RT-PCR and flow cytometry, respectively. The mRNA and protein expression of peroxidase proliferator-activated receptor(PPAR β) were detected with RT-PCR and Western blot, respectively. Results:After 3-months rosuvastatin treatment, the patients' low-density lipoprotein cholesterol (LDL-C) levels decreased significantly (P<0.01). Compared with baseline, the mRNA and protein expressions of CCR2 in the mononuclear cells showed significantly decrease, as well as plasma MCP-1 levels (P<0.05). Both mRNA and protein expressions of PPAR β in the mononuclear cells increased (P<0.05). Conclusion:Rosuvastatin may attenuate MCP-1/CCR2 through PPARβ upstream pathway.

Objective:To investigate the inotropic effects of dioscin (Dio)in rat isolated-heart and intracellular free calcium concentration in isolated rat ventricular myocytes and to explore its mechanism preliminarily. Methods:Left ventricle contractile function was measured using the Langendorff non-recirculating mode of isolated rat heart perfusion. Effects of dioscin and dioscin+SEA0400, sodium-calcium exchanger (NCX) inhibitor, were investigated by measuring left ventricular systolic pressure (LVSP) and left ventricular end diastolic pressure (LVEDP). Also, heart rate (HR), peak rate of rise/fall of left ventricular pressure (±dp/dt_max) of isolated rat heart were calculated; Effects of dioscin and SEA0400 on intracellular free calcium concentration in rat H9c2 cells were measured by Fluo3-AM and then detected the fluorescence intensity with confocal microscopy. Results:With 1 μmol/L dioscin, LVSP was significantly increased by 19.7% (P<0.01) and dp/dt_max was increased by 9.6%; With 1 μmol/L dioscin, the relative fluorescence intensity of intracellular free calcium concentrations were strong significantly(P<0.01). While in presence of SEA0400, the relative fluorescence intensity was changed to 17.09±0.63 (P<0.01) by 1 μmol/L dioscin. With 1 μmol/L dioscin, the relative fluorescence intensity was weak(P<0.01) without calcium or sodium in the extracellular fluid. Conclusion:Dioscin shows positive inotropic effect on isolated rat heart, enhancing the LVSP and +dp/dt_max; Dioscin increases the intracellular concentration of Ca²⁺ in the cardiac myocytes by increasing Na⁺ influx and facilitating the reverse mode of the sodium-calcium exchanger.

Objective:To investigate the roles of catestatin (CST) in 2-kidney-1 clip (2K1C)-induced renal hypertension in rats, and to explore the underlying mechanism. Methods:Thirty six male SD rats were randomly divided into Sham group (n=15) and Model group(n=21).The model group was performed by 2K1C operation. For 2K1C operation, the left renal arteries were narrowed by cotton thread. The Sham group was treated with the same condition as the 2K1C group except the renal artery was narrowed. Tail-cuff systemic blood pressure of rats was measured before and every weeks after 2K1C operation. Six weeks after 2K1C operation, a carotid artery catheter was inserted to measure blood pressure of rats under anesthesia. Then, the model group was randomly subdivided into 2K1C group (n=15) and 2K1C+CST group (n=6). The rats of 2K1C+CST group were intravenous given CST (80 μg/100 g) and the rats of Sham or 2K1C group were given normal saline. All rats were sacrificed after blood pressure was measured and blood was collected. Then, the left ventricular plus interventricular septum weight (LV+S) was weighted and the ratio of (LV+S)/body weight(BW) was calculated as the index of left ventricular hypertrophy. Norepinephrine (NE) contents in plasma were determined by high performance liquid chromatography(HPLC) and CST contents in plasma by ELISA. The nitrite/nitrate contents in the left ventricular tissue and plasma were measured by nitrate reduction method to represent nitric oxide (NO)contents.Expression levels of CST in the left ventricle, kidney, medulla oblongata and adrenal gland,as well as eNOS and iNOS, were tested by Western blot. Results:①The 2K1C group had higher tail-artery blood pressure(P<0.01) and were more marked presence of right ventricular hypertrophy than those of sham group (P<0.01). Compared with Sham group, plasma CST content in 2K1C group was decreased by 226% (P<0.01), while plasma NE content in 2K1C group was increased by 246% (P<0.01), expression levels of chromograminA(Chga) in medulla oblongata of 2K1C group were increased by 108%, in leftventricle and kidneywere decreased by 60% and 30%, respectively (P<0.05).the content of NO in left ventricular and plasmawere increased by 46% and 24% respectively. ②The carotid arterial blood pressure of 2K1C group markedly reduced after administration of CST.③Compared with 2K1C group, the content of NO in left ventricul and plasma of 2K1C+CST group were increased by 35% and 19% respectively(P<0.05). The expression of eNOS in left ventricular of 2K1C+CST group were also obviously increased. Conclusion:The CST expression of 2K1C-induced renal hypertension rats is reduced and the effects of exogenous CST lowering their blood pressure may be related to NO/NOS system.Therefore, we speculate CST could contribute to the pathogenesis and progression of renal hypertension.

Objective:To observe the protective effect of irbesartan on myocardial fibrosis in diabetic rats, and analyze the role of extracellular signal-regulated kinase (ERK) pathway in this protection. Methods:Thirty two male SD rats were randomly divided into two groups:normal control group (CON, n=10), experimental group (n=22). Twenty diabetic rats which had modelled successfully were randomly divided into two groups:diabetic group (DM, n=10), irbesartan + DM group (Ir+DM, n=10). After 8 weeks, fasting blood glucose (FBG) level, body weight (BW), the ratio of heart weight/body weight (H/B) and left ventricular weight index (LVWI) were measured. The myocardial morphological and fibrotic changes were observed by Masson staining. Col I and col Ⅲ contents were evaluated using ELISA method, and the protein expressions of ERK1/2, p-ERK1/2 in heart tissue were tested by Western blot. Results:Compared with CON group, in diabetic rats, the levels of FBG, H/B and LVWI were increased while BW was decreased. The contents of col I and col Ⅲ were increased as well as the protein expression of p-ERK1/2 and the ratio of p-ERK1/2/ERK1/2(P<0.05,P<0.01), which had the statistic differences, while ERK1/2 protein expression was not changed. Masson staining showed myocardial collagen was increased, arranged in disorder and uneven distribution. However, in Ir+ DM group, BW was increased obviously, H/B, LVWI, the contents of col I and col Ⅲ were decreased significantly (P<0.05,P<0.01), p-ERK1/2 protein expression and the ratio of p-ERK1/2/ERK1/2 were decreased (P<0.01), which had the statistic differences. Meanwhile myocardial morphology was improved significantly. Conclusion:Diabetes can induce the happening of myocardial fibrosis, and irbesartan can induce the damage of myocardial fibrosis through inhibitting the activation of ERK.

Objective:To investigate the effects of endurance training on interleukin-18(IL-18) and IL-10 in atherogenice apolipoprotein E gene knockout(ApoE^-/-)mice and explore its possible mechanism preventing atherosclerosis occurrence. Methods:Twenty male ApoE^-/- mice (age:8 weeks) were randomly divided into atherosclerosis(AS)model group(AC) and exercise group(AE) and chose male C57BL/6J mice (age:8 weeks) as control group (CC). After 12 weeks, the aortic trunk closed heart was prepared for frozen section, which were used to observe the changes of plaque area and pathology. The expressions of IL-18 and IL-10 proteins and the levels of blood IL-18 and IL-10 were measured by ELISA. Results:① Twelve weeks high fat diet significantly resulted in typical AS lesion, however, endurance training markedly decreased AS plaque area and pathology in ApoE^-/- mice(P<0.01). ②Compared with CC group, serum IL-18 and IL-10 levels were significantly increased in AC group and AE group and IL-18/IL-10 ratio was also increased markedly in AE group(P<0.01). Serum IL-18 level and IL-18/IL-10 ratio in AE group were all significantly lower than those in AC group (P<0.01).③Compared with CC group, the expressions of IL-10 and IL-18 proteins in aorta were significantly increased in AC group and AE group (P<0.01). The expression of IL-10 protein in aorta in AE group was markedly higher than that in AC group but IL-18 protein expression was significantly lower. Conclusion:Endurance training can prevent AS appeared through enhancing aortic anti-inflammatory capability by decreasing serum and aortic IL-18 levels and by increasing serum and aortic IL-10 level.

Objective:To discuss the modulating effects of shenmai (Traditional Chinese Medicine) injection on blood lipid in hyperlipidemia rats through observing the serum lipid level and a series of related biochemical indexes in hyperlipidemia rat model. Methods:A total of 30 male SD rats of SPF grade were purchased and fed with basic feed for 1 week to adapt the environment. Then the rats were randomly divided into the following groups(n=10):control group, model control group, shenmai injection group. Control group was fed basal diet; the latter two groups were fed high fat diet,the body weight of all the animals was measured each week. For shenmai injection group, the rats were fed with shenmai injection (10 ml/kg) twice a day for 45 consecutive days through oral administration. The effects of shenmai injection on body weight, serum triglyceride (TG), total cholesterol (TC), low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C), super oxide dismutase (SOD), glutathione peroxidase(GSH-Px), malondialdehyde (MDA), the activities of lipoprotein lipase(LPL) and hepatic lipase (HL) in hyperlipidemia rats were detected. Results:Body weight, serum of TC, TG, LDL-C, HDL-C, and MDA levels in model control group were significantly higher(P<0.01), while serum of HDL-C, SOD, GSH-Px, LPL, and HL levels were significantly lower than those of control group(P<0.01). Body weight, serum of TC, TG, LDL-C, HDL-C, and MDA levels in shenmai injection group were significantly lower(P<0.01), while serum of HDL-C, SOD, GSH-Px, LPL, and HL levels were significantly higher than those of model group(P<0.05, P<0.01). Conclusion:Shenmai injection has a significant effect of modulating blood lipid and antioxidant function on hyperlipidemia rats.

Objective:To study the effect of acute cold exposure on the expression of aquaporin-1 (AQP-1) and AQP-5 in lung tissues and the changes of ultrastructural pathological changes after cold exposure in rats. Methods:Twelve male Wistar rats were randomly divided into control group (23±2)℃ and cold (-25℃) exposure group, the exposure time was 2 h. Rectal temperatures of the rats were measured immediately after cold exposure. The ultrastructural pathological changes of pulmonary tissue were observed by transmission electronic microscope. The mRNA expression of AQP-1 and AQP-5 was measured by RT-PCR. The protein expression of AQP-1 and AQP-5 was measured by Western blot. Results:The body core temperatures(28.07±4.15)℃ of the cold exposure group were decreased significantly compared with the control group(37.33±0.25)℃ (P<0.05). In acute cold exposure group, the main pathological changes of pulmonary ulstructure included pulmoary epithelial basement membrane thickening, and karyopyknosis of AT-I cells, and vacuolization on the cytoplasm of (AT-Ⅱ) cells. After acute cold exposure, the levels of both mRNA and potein expressions of AQP-5 were decreased significantly (P<0.05) compared with those in the control group. while AQP-1 expression level showed no statistical significance between control group and cold exposure group. Conclusion:There might be some cause and effect relationship between lung tissue damage by cold exposure and the levels of mRNA and potein expressions of AQP-5 decreased after acute cold exposure.

Objective:To study the dilatation characteristics of ATP-sensitive potassium channel (K_ATP) SUR2B/Kir6.1 subtype opener iptakalim (Ipt) in pulmonary arterioles, and to explore its possible mechanism. Methods:Vessels pressure-diameter monitoring perfusion technique was used to observe the dilatation effects of Ipt in rats fourth pulmonary arterioles (n=6~8). After the pulmonary arterioles were pre-treated with removing endothelium or pre-incubated with K_ATP channel blocker glibenclamide (Gli), cyclo-oxygenase (COX) inhibitor indomethacin (Indo) and nitric oxide synthase (NOS) inhibitor L-Nω-Nitro-arginine methyl ester(L-NAME), the dilatation effects of Ipt were observed. Results:Pulmonary arterioles could be relaxed by Ipt, the maximal relaxation rate was (60.53±2.08)%. Compaired with control group, the effects of Ipt in endothelium denuded arterioles were significantly decreased, the maximal relaxation rate was (9.47±1.56)% (P<0.01). The maximal relaxation rate were decreased to(17.49±1.47)%,(37.00±3.88)% and(24.91±2.30)% respectively after Gli,Indo,L-NAME were pre-incubated (P<0.01). Conclusion:Pulmonary arterioles can be relaxed by Ipt. The selective activation of K_ATP SUR2B/Kir6.1 subtype by Ipt was involved in its mechanisms. The endothelium-dependently dilatation of Ipt was related to nitric oxide (NO) and prostacyclin (PGI₂) released by endothelial cells.

Objective:To investigate the effects of extracts from Ajuga decumbens on anti-fatigue in mice. Methods:One hundred and twenty female Kunming (KM) mice were randomly divided into quiet control group, sport control group, positive control group and 3 experimental groups which were the low, medium and high dose group given the extracts from Ajuga decumbens. The low, medium and high dose group were given the extracts with 100 mg/kg, 200 mg/kg, 400 mg/kg by body weight of mice for 30 d, respectively, but the positive control group was given American ginseng granules, while the quiet control group and the sport control groups were treated with saline. After this, the exhausting time, the physio-biochemical indexes (including lactic acid, blood urea nitrogen, blood sugar, total cholesterol and triglyceride) in serum, the contents of muscle and liver glycogen, and the antioxidative indexes (including glutathione peroxidase, superoxide dismutase, catalase and malondialdehyde) of organs in mice were investigated. Results:The exhausting time, the number of red blood cell, the contents of hemoglobin and blood sugar, the contents of muscle and liver glycogen, and the activities of glutathione peroxidase, superoxide dismutase and catalase in organs of mice in the medium dose group and the high dose group were significantly more than those of the sport control group, but the contents of blood lactic acid, blood urea nitrogen and that of triglyceride and total cholesterol in serum, and the content of malondialdehyde in organs of mice in the medium dose group and the high dose group were significantly lower than those of the sport control group. And the effect of medium dose extracts from Ajuga decumbens on anti-fatigue was better than that of American ginseng granules. Conclusion:The extracts from Ajuga decumbens has significant anti-fatigue effect in mice.

Objective:To explore the role of Nrf2/ARE pathway in skeletal muscle ischemia/reperfusion(I/R) injury preconditioning by dexmedetomidine(DEX). Methods:Twenty-eight SD rats were randomly divided into sham-operated(Sham group)、I/R group、I/R+ DEX(DEX group) and I/R+DEX +Atipamezole (Atip group). In the Atip group, Atip(250 μg/kg) and DEX(25 μg/kg) were injected together after anesthesia; In the Sham and I/R groups, the homologous saline was also injected at the same time; In the DEX group, the homologous DEX and saline were coinjected. After 30 minutes, the hind limb ischemia was induced by clamping the common femoral artery and ligaturing collateral circulation. After 3 h of ischemia, the clamp and tourniquet were removed and the rats underwent 2 h of reperfusion. We measured plasma concentrations of lactate dehydrogenase (LDH) and creatine kinase(CK). The gastrocnemius muscle was harvested and immediately stored at -80℃ for the assessment of malondialdehyde(MDA)、superoxide dismutase(SOD) and Nrf2/HO-1 protein detected by Western blot. The other section muscle was stored in triformol for immunohistochemical and HE staining. The wet/dry was also immediately detecting. Results:The levels of wet/dry、MDA、LDH、CK、Nrf2 and HO-1 were higher(P<0.05) while the level of SOD was lower(P<0.05) in the I/R group than those in sham group. The levels of wet/dry、MDA、LDH、CK were significantly lower(P<0.05) yet the levels of SOD and Nrf2/HO-1 were significantly higher(P<0.05) in DEX group than those in I/R group. However, Atip reversed the effect of DEX in Atip group, each of indicators had significant changes compared with those in the DEX group(P<0.05). Conclusion:Nrf2 protein was expressed in skeletal muscle of rat and DEX could promote its level in nucleus by α-adrenergic receptor. The down-stream products of Nrf2 have the effect of antioxidant.

Objective:To analyze the combined features of serum biomarker of exercise-intervened type 2 diabetes mellitus (T2DM) rats at different time point. Methods:Ninety SD rats randomly selected from 100 were fed with high glucose and fat diet for 4 weeks, and then received intraperitoneal injection of streptozotocin(STZ) according to 40 mg/kg by weight; after modeling for one week, Seventy-one rats were successfully modeled among which intervened with swimming at three different time points (2w, 4w and 8w). Eighty-one rats were finally divided into 8 groups as below:Normal control group (C₀,n=10);DM model comparison group (DC₀,n=10); DM model comparison 2w group (DC₂, n=11); DM model comparison 4w group (DC₄, n=10); DM model comparison 8w group (DC₈, n=9); DM exercise intervention 2w group (DE₂, n=11); DM exercise intervention 4w group (DE₄, n=10); DM exercise intervention 8w group (DE₈, n=11).The UPLC/Q-TOF MS technique was used to conduct metabonomics test of serum of rats to analyze the combined features of serum biomarkers under exercise intervention at different time points. Results:According to fold change, we got the biomarker combinations (with 15 specific substances in each combination) of exercise intervention at three time points. In these serum biomarker combinations that were specific at different time points, a majority of metabolites appeared simultaneously at different time points but with obvious difference of fold change. Fold change of monoglyceride(24:6) and gluconic acid were most evident respectively at 2w and 4w intervention time points,Eicosapentaenoic Acid (EPA) andlinolenic acid(LA) contents might approach the normal level at 4~8 time point. Conclusion:Combined features of serum biomarker of exercise-intervened T2DM rats are specific at different time point. Both EPA and LA are common substances significantly up-regulated while MonoglycerideMG(24:6), Gluconic acid, Propionyl-L-carnitine(PLC), Arginine(Arg) and Sphingosine-1-phosphate (SPP) are common substances significantly down-regulated at three time points.

Objective:To explore the possible biological mechanisms of skeletal muscle injury recovery by observing the changes of AMP-activated protein kinase α2 (AMPKα2) and hypoxia inducible factor-1α (HIF-1α) expression levels of rats in the skeletal muscle blunt injury recovery process. Method:Six of the 48 male Wistar rats was randomly selected as the normal control group. The blunt injury model was set up by injuring the hind legs of remaining 42 rats with heavy objects. Then they were divided into 7 groups (n=6). The changes of AMPKα2 and HIF-1α expression levels were tested in the hind limb triceps surae of the model rats from each of the 7 groups at 12 hours, 2 days, 5 days, 7 days, 10 days, 15 days and 30 days respectively following the injury. Results:The expression levels of AMPKα2 and HIF-1α all increased significantly at 12 hours following the injury and fell close to normal levels at 15 days following the injury. The values of AMPKα2 and HIF-1α expression peaked within 2 days after injury and the expression levels began to decline at 5 days after injury. Except the peak, the changes of the mRNA expressions of the two proteins were basically consistent with those of protein expression at the other time points. Conclusion:HIF-1α and AMPKα2 might play roles in mediating hypoxia adaptation, muscle cell regeneration, and energy compensation to promote recovery after injury.

Objective:To study the change of airway mucus secretion under a high temperature and humidity environment, and explore the effects of hot-humid stress and acclimation on the morbidity and mortality of respiratory disease. Methods:Forty-five BABL/c mice were randomly divided into five groups:normal group, hot-humid group I, hot-humid group Ⅱ, hot-humid group Ⅲ, hot-humid group IV, with 9 mice in each group. Mice in normal group were continuously placed in the common environment and sacrificed after 7 days. Mice in other groups were housed in a temperature-and-humidity-controlled environment (33℃±0.5℃, 95%±5%). Mice in hot-humid group I, hot-humid group Ⅱ, hot-humid group Ⅲ and hot-humid group IV were sacrificed after 12 hours, 24 hours, 4 days, 7 days respectively. The protein expression of mucin 5AC(MUC5AC)、epidermal growth factor receptor (EGFR)、aquaporin 1(AQP1) and aquaporin 5(AQP5) in lung were tested by immunohistochemisty. Results:After housed in a high temperature and humidity environment, immunohistochemisty revealed a significant increase of AQP5 12 h later, MUC5AC and EGFR 24 h later, compared with normal group(P<0.05). There was a significant decrease of MUC5AC 7 d later, compared with normal group(P<0.05). There was no significant difference in MUC5AC, EGFR and AQP5 expression among all groups at other time points. There was no difference of AQP1 in humid heat groups, compared with normal group, but a significant decrease in humid heat Ⅲ and IV groups, compared with humid heat I and Ⅱ groups. Conclusion:These findings indicate that hot-humid stress induces mucus hypersecretion in airways, which may be related to the up-regulation of EGFR and down-regulation of AQP5 in MUC5AC. Although acclimation mitigates above-mentioned response, a series of more complex responses may be induced if still in the hot-humid environment.

Objective:To examine whether agmatine (AGM) would alter stress-induces hyperthermic response. Methods:Sixty-one male SD rats were randomly divided into three experiments. Each experiment was divided into control group and AGM group. During the experiments, the animals were maintained in a chamber at 22℃. ①Effects of intraperitoneal injecting 40 or 80 mg/kg AGM on normal core temperature and activity were observed in undisturbed rats using radiotelemetry (n=8). ②Stress-induced hyperthermia model was established by placing rats in an open-field chamber for 60 min. Rats were dosed intraperitoneally with AGM or saline, and placed immediately inside the open-field chamber. Core temperature and motor activity were monitored by radiotelemetry in an open-field chamber (n=7~8). ③Effect of AGM on energy metabolism was measured by Columbus Oxymax Lab Animal Monitoring System (n=7). Results:①Rats administered with 80 mg/kg AMG showed significant hypothermic responses (-0.46±0.11)℃, while 40 mg/kg AMG had no significant effect on the normal core temperature. ②Core temperature of control group increased by (0.78±0.16)℃ during open-field exposure, whereas rats administered 40 and 80 mg/kg AGM underwent a (0.34±0.11)℃ and (0.81±0.14)℃ reductions in core temperature within 60 min, respectively. ③Oxygen consumption and energy metabolism were significantly reduced by AGM (80 mg/kg). Conclusion:The data demonstrated that AGM induced hypothermic responses in rats and reversed stress-induced hyperthermia, and its effect might attribute to the suppression of energy metabolism.

Objective:To study the effect of excited dopamine type I receptor on the production of nitric oxide/nitric oxide synthase(NO/NOS)in ox-LDL activated THP-1 cells and the possible mechanism. Methods:Cultured THP-1 cells activated by PMA were randomly assigned in the following groups:control group (control), oxidized low density lipoprotein group (ox-LDL), dopamine receptor 1(DR1) agonist group (SKF), DR1 antagonist group (SCH), ERK blocker group (PD98059). Oil Red O staining was used to identify the accumulation of cellular lipid. The levels of NO and NOS in the supernatant of THP-1 were assayed by nitrate reductase method. The protein expression of DR1, p-ERK and ERK were obtained by Western blot and immunity fluorescence. Results:After 48 h of incubation of ox-LDL, accumulation of lipid in the cytoplasm was found in most THP-1 cells. Compared with control group, DR1 protein expression was reduced in ox-LDL-induced cells(P<0.01). Activation of DR1 agonist decrease the production of NO and iNOS(P<0.01), and PD98059 partly reversed the above effect. Conclusion:Activation of DR1 can inhibit the production of NO/NOS in ox-LDL-induced THP-1 cells, which may be related with ERK pathway.

Objective:This study aimed to construct a large conductance calcium activated potassium channel α (BKCa) subunit plasmid with two tags by the overlapping PCR technique to set up a steady base for future ion channel study. Methods:Based on the existing coding BKCa channel α subunit expression plasmid pcDNA3.1-hSlo, we constructed a double-tag expression plasmid, namely, pcDNA3.1-Flag-hSlo-GFP (Flag-hSlo-GFP). Results:Flag tag was inserted into the S1-S2 extracellular loop of BKCa channel α subunit, and GFP tag was connected to the C-terminus of BKCa channel α subunit. Sequence of the constructed plasmid was confirmed successful. Conclusion:The expression plasmid Flag-hSlo-GFP was constructed successfully with overlapping PCR. Overlapping PCR is a valuable method for amplifying long size genes.

Objective:To bring about physiological researchers' attention of the importance of sample size estimation. Methods:The significance as well as the current problems of sample size estimation were illustrated and the commonly-used sample size estimation methods were introduced. Results:The basic concepts and necessary premises of sample size estimation were stated. The estimation processes and results under two different circumstances were elaborated in detail via examples. Conclusion:To attain the proper estimated sample sizes, the computation must satisfy the necessary premises which included the appropriate statistical analysis methods to be used.