BACKGROUND:Insulin-like growth factor-1 (IGF-1) is an important mitogen and an indispensable regulator during normal hair cell development. Extracellular signal regulated kinase 1/2 (ERK1/2) is also expressed in mammalian vestibular organs.OBJECTIVE: To investigate the changes of the expression and the distribution of IGF-1, IGF-1 receptor (IGF-1R) and ERK1/2 in guinea pig's vestibular epithelium following gentamycin injury.DESIGN: A randomly controlled study.SETTING: Department of Otorhinolaryngology, General Hospital of Chengdu Military Area Command of Chinese PLA.MATERIALS: Twenty healthy and adult guinea pigs, weighing 300 to 350 g, provided by the Experimental Animal Center,Fourth Military Medical University of Chinese PLA, were randomly divided into four experimental groups and a control group with 4 in each group.METHODS: The experiment was performed at Department of Otorhinolaryngology Research Laboratory of Xijing Hospital,Fourth Military Medical University of Chinese PLA from January 2002 to May 2002. In the experimental groups,gentamycin was intraperitoneally administered daily (80 mg/kg) for 10 consecutive days. For the control group, 1 mL of 0.9% sterile saline was administered in the same way. The guinea pigs, in which unsteady gait or nystagmus appeared,could be involved in the further experiments. Before being sacrificed, the animals' gait, nystagmus and the amount of food-intake were observed every day. The animals were anesthetized deeply with 50 mg/kg pentobarbital 1, 7, 14 or 21days after the last gentamycin administration and then decapitated. Bilateral otic vesicles were harvested and immersed in 4% paraformaldehyde. Then the specimens were decalcified in 10% ethylenediamine tetraacetic acid (EDTA),thereafter embedded in OCT gel and serially sectioned at a thickness of 10 μm in the cryostat. The behavioral changes of guinea pigs were observed. The morphological changes of vestibular epithelium were examined by hematoxylin and eosin (HE) staining. The expression and distribution changes of IGF-1/1R and ERK1/2 were investigated immunohistochemically. MAIN OUTCOME MEASURES: The behavioral, morphological, expression and distribution changes of IGF-1/1R and ERK1/2 of guinea pigs in each group.RESULTS: Twenty involved guinea pigs entered the stage of final analysis. ① All the guinea pigs' vestibular functions were impaired after gentamycin treatment, then partially restored without any pharmaceutical treatment. ②Morphological injury was obvious after gentamycin treatment, and improved 1 week later. ③The expression of IGF-1/1R was low in the control group, and in the 1-day group it increased significantly to its maximum. After that, its expression decreased gradually, but was still higher in the 21-day group than that in the control group. There were significant differences in the expression of IGF-1/1R among different groups (F =51.8,45.7,P ＜ 0.05). The expression changes of IGF-1 and its receptor were similar. ④The expression of ERK1/2 was low in the control group, and increased gradually after gentamycin toxicity. In the 7-day group, the immunoreactivity reached its maximum. Then the levels of ERK1/2 decreased gradually, but were still higher in the 21-day group than that in the control group. There were significant differences in the expression of ERK1/2 among different groups (F =103.7,106.4, P ＜ 0.01), but no significant differences existed in the expression between ERK1 and ERK2 among different groups (P ＞ 0.05).CONCLUSION: The expression of IGF-1/1R and ERK1/2 in vestibular epithelium increased after gentamycin treatment,and IGF-1 may play an important role as an endogenous mitogen through a paracrine or autocrine manner in the early stage of hair cell self-repair of guinea pigs. ERK1/2 may play an important role in signal transduction during vestibular hair cell self-repair after gentamycin toxicity in guinea pigs.

OBJECTIVETo study orientation remodeling without stress in bone harvest chamber.

METHODSThe bone harvest chamber (BHC) methodology is adopted in this study. Five female Japanese white rabbits were allowed unrestricted activity. The bone harvest chamber was a cylindrical Ti implant body with a transverse 1 mm wide canal for bone ingrowths. Retrieval of the contents of the canal was allowed with minimal disturbance to the surrounding bone or outer cylinder. After bone harvest chambers were implanted into the tibia of rabbits for 8 weeks, the chambers were considered to be osseointegrated with the bone. After harvested, the tissue were fixed and decalcified, then embedded in paraffin. Each rabbit was put into surgical operation 4 times for 4 stages: vacant for the first time; the tissue were cut into longitudinal sections at the second and third stages; harvesting tissues were cut into transverse sections at the fourth stage. Directional analysis: the standard deviation of the orientation of cell nucleus in each section was used as statistics, the difference between longitudinal section and transverse section were analyzed.

RESULTSOf the tissue into bone harvest chamber, directionality of cells arranged was more significantly on longitudinal section than on transverse section and there was statistical ignificecne.

CONCLUSIONUnder the no-stress circumstance of BHC bone remodeling showed directivity. Stress is not the direct leading signal about bone reconstitution. The structure of BHC might be related to orientation remodeling, which suggests that the relationship between orientation and stress is mediated by blood vessel. The effect of stress may be to affect vessel distributing in some orietation.

Animals ; Biomechanical Phenomena ; Cell Culture Techniques ; instrumentation ; methods ; Cells, Cultured ; Female ; Osteocytes ; chemistry ; cytology ; Rabbits ; Tibia ; chemistry ; cytology

AIM: To establish an experimental model of high intraocular pressure in mice by laser photocoagulation and to prepare for future research.
METHODS: Experimental model of high intraocular pressure was induced unilaterally in 44 C57BL/6 mice. The fellow eye served as a control. TONO-PEN AVIA Tonomter was used to measure intraocular pressure (IOP) to guarantee IOP value at 1, 2, 4, 8wk. Slit-lamp biomicroscopy was performed throughout the period and the structural changes were assessed histologically. And then, their eyes were enucleated, postfixed, cryoprotected, and embedded in optimal cutting temperature medium. After hematoxylin and eosin stain ( HE stain ) , cryosections of the retina were observed under light microscope. TdT-mediated biotin-dUTP nick end labeling ( TUNEL ) was performed on the retinal sections to determine apoptosis rate.
RESULTS: IOP of laser-treated eyes was significantly higher than that of control eyes from 1-8wk (P<0. 05). The highest IOP was 31mmHg, but only one eye. The IOP was mainly around 20mmHg. In laser-treated eyes, the angle of anterior chamber were narrow. Number of cells in the inner nuclear layer and retial gangllion cell layer was slightly lower than that in control eyes at 2wk, but by 4 and 8wk the number of cells was significantly lower than that in the control contralateral eyes.
CONCLUSION: The laser photocoagulation of limbus causes chronic elevation of IOP and this method may be a promising experimental model for the investigation of biological mechanisms of glaucomatous retinal ganglion cell damage.

OBJECTIVETo provide a scientific basis for systematic research on the mechanism of chronic immobilization stress (CIS) induced metabolic network change in rats, through detecting the changes of endogenous metabolites in rats with CIS, treated or un-treated with Xiaoyao Powder (XYP), for determining the small molecule marker compound that closely associated with the metabonomical specificity of CIS and acting mechanism of XYP.

METHODSThirty-six experimental male SD rats were divided into 3 groups, the normal control group, the model group and the XYP group. And all the three groups were subdivided into two subgroups respectively on day 7 and day 21 of the experiment. The stress rat model of CIS was made by chronic restraining method for 3 h every day. Starting from the first day of modeling, XYP 3.854 g/kg in volume of 1 mL/100 g body weight was administered 1 h before restraining via gastrogavage to rats in the XYP group, while equal volume of distilled water was given to rats in the other two groups instead. Blood samples were collected on the 8 th day and 22 th day for detection in the following procedure: at 27 degrees C, 300 microL supernate of blood plasma was taken, calling the Carr-Purcell-Meiboom-Gill (CPMG) and longitudinal eddy-delay (LED) sequence respectively on a Fourier variable nuclear magnetic resonance (NMR) spectrometer, pre-saturated inhibition of the water peak was performed; free induction decay (FID) signals were transferred via 32 k Fourier transformation to gain one-dimensional NMR spectrogram; by taking TSP as the chemical migration reference peak, the segmental integral calculus (0.04 ppm per segment) was performed from 4.5 - 0.5 ppm (CPMG) and 6.0 - 0 ppm (LED) within the peak ranges in 1H spectra using the VNMR software; after normalization, centering and scaling were conducted on data, then used for pattern recognition of principal component analysis (PCA) using the SIMCA-P 10.0 software, or if necessary, the partial least squares discriminate analysis (PLS-DA) was performed.

RESULTS(1) The metabolites in the model group were significantly different from those in the control group, suggesting that the animal model was successfully established with the metabolic network different to that of control. The model group and the XYP group could be differentiated from the control group by the differences of metabolites and metabolic networks between groups; XYP could intervene the metabolites or the metabolic path to cause changes in final metabolites. (2) The serum contents of lactic acid (1.4, 4.16), choline (3.24), N-acetylgalactosamine (NAC) and saturated fatty acids (1-3) increased, but unsaturated fatty acids (1.99,4-5), blood sugar (34), HDL (0.83), etc. reduced in the CIS rats. XYP showed obvious regulatory effects on final metabolites, causing decrease of lactic acid, choline, NAC, saturated fatty acids and blood sugar, and increase of unsaturated fatty acids, blood sugar, HDL, 3.44 ppm compound, etc.

CONCLUSIONSThe metabolic phenotype in CIS rats includes the increase of lactic acid, choline, NAC, saturated fatty acid, and the decrease of blood sugar contents, unsaturated fatty acid, HDL, 3.44 ppm compound, etc., these may be the markers of the metabolites. The final metabolites changes induced by CIS are primarily the lipid substances. XYP markedly regulates the contents of final metabolites, showing the regulatory effects on final metabolites, but what is the metabolite or metabolic pathways it interferes to alter the final metabolites should be confirmed by further studies.

Animals ; Blood Chemical Analysis ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Humans ; Magnetic Resonance Spectroscopy ; Male ; Metabolomics ; Powders ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Stress, Physiological ; drug effects

Chemogenomics/chemical genomics has been widely used in novel drug research and discovery. Firstly, the concept of chemogenomics was introduced briefly. Secondly, we reviewed the progress of novel drug research and discovery based on forward chemogenomics, reverse chemogenomics and predictive chemogenomics. Finally, we showed progress of the research that chemogenomics has been used to novel drug research and discovery in pharmaceuticals companies.
Animals ; Drug Delivery Systems ; Drug Discovery ; methods ; Drug Industry ; trends ; Genomics ; methods ; Humans ; Pharmacogenetics ; methods

OBJECTIVETo evaluate whether or not administration of folic acid and resveratrol have preventive effects on cleft palate formation as well as the comparison of the two drugs' s effects.

METHODSPregnant mice were randomly divided into 9 groups, with 8 mice in each group. The TCDD group mice were dosed with TCDD 28 microg/kg body weight on gestation day 10 (GD 10) animals in folic acid group were respectively dosed with folic acid 15, 10, 5 mg/kg and TCDD 28 microg/kg; resveratrol treated mice were divided into 3 groups: resveratrol 50 mg/kg were orally administered for 6 consecutive days, from gestational day GD 8 to GD13 in resveratrol (GD8-13 ) group; resveratrol 50 mg/kg were orally administered for 6 consecutive days, from gestational day GD 8 to GD13, followed hy an oral administered with TCDD on GD10 in resveratrol (GD8-13) + TCDD group; resveratrol 50mg/kg and TCDD 28 microg/kg were used by gavage administration at GD10 in resveratrol (GD10) + TCDD group. Control mice were treated with the same volume of water for 6 consecutive days from GD8 to GD13 and were given a single dose of corn oil on GD10. The pregnant mice weight and embryos, the number of live, cleft palate, dead and resorption fetal mice were recorded on GD 17.5. The coronal sections of the fetal mice heads were prepared at GD 17.5 and observed by microscopy.

RESULTSTotal frequency of clefts was 92.86% in TCDD group, 84.00% (15 mg), 73.08% (10 mg), 84.00% (5 mg) in folic acid + TCDD groups, 0% in resveratrol (GD10) group, 74.51% (GD10), 57.78% (GD8-13) in resveratrol + TCDD groups. The frequency of cleft was 0% in the control group. Compared with the control and the TCDD groups, there were significant differences in the number of live, dead and resorption fetal mice in TCCD + resveratrol (GD8-13) group (P < 0.05). No significant differences in embryonic weight, live fetuses weight, the number of live, dead and resorption fetal mice were found in the other groups (P > 0.05).

CONCLUSIONTest dose of folic acid and resveratrol both had certain antagonistic effect on cleft palate in mice induced by TCDD, with folic acid 10 mg/kg, resveratrol 50 mg/kg GD8-13 doses having stronger antagonistic action. Effects of both the two drugs have no significant difference, but resveratrol (50 mg/kg, GD8-13) significantly affects the fetal mice's growth and development under TCDD exposure in utero.

Abnormalities, Drug-Induced ; prevention & control ; Animals ; Cleft Palate ; chemically induced ; prevention & control ; Female ; Fetus ; Folic Acid ; administration & dosage ; pharmacology ; Humans ; Mice ; Mice, Inbred C57BL ; Polychlorinated Dibenzodioxins ; antagonists & inhibitors ; Pregnancy ; Random Allocation ; Stilbenes ; administration & dosage ; pharmacology ; Teratogens

To research the effect of Ginseng Radix et Rhizoma and Aconiti Lateralis Radix Praeparata compatibility on cardiac toxicity in rats by UPLC-Q-TOF/MS, and explore the endogenous markers and molecule mechanism. Different compatibility of Shenfu decoction were given to male Wistar rats at dosage of 20 g · kg(-1) for 7 days, collected the serum, and analyze the endogenous metabolites effected by Shenfu formulation by principal component analysis and partial least-squares analysis. Results showed that content of glutathione, phosphatidylcholine and citric acid decreased in mixed-decoction group, while ascorbic acid, uric acid, D-galactose, tryptophan, L-phenylalanine increased. The results showed cardiac toxicity of Aconiti Lateralis Radix Praeparata in Shenfu mixed-decoction. Shenfu co-decoction group showed a similar or weaker trend compared with control group, but most of them do not have a statistically significant. The results indicated the scientific basis of Shenfu compatibility by comparison of co-decoction group with mixed-decoction group. Shenfu compatibility can reduce cardiac toxicity induced by Aconiti Lateralis Radix Praeparata, and citric acid, glutathione, phosphatidyl choline, uric acid might be regarded as potential markers of cardiotoxicity.
Animals ; Biomarkers ; Cardiotoxicity ; Drugs, Chinese Herbal ; toxicity ; Glutathione ; blood ; Least-Squares Analysis ; Male ; Metabolomics ; methods ; Principal Component Analysis ; Rats ; Rats, Wistar

OBJECTIVETo study the correlation factors of acute paraquat intoxication prognosis.

METHODSThe early paraquat concentration in plasma and urine, leukocyte count, hepatic and renal function, amylase, electrolyte and the parameters of arterial blood gas were analyzed retrospectively in 111 patients with acute paraquat intoxication.

RESULTS43 cases (38.7%) of all the 111 patients survived and the other 68 cases (61.3%) died. The patient, whose paraquat concentration was not more than 8.0 µg/ml in plasma and 276.0 µg/ml in urine, could survive. But some patients could die, only if there was no paraquat found in plasma. The paraquat levels in plasma and urine were significantly lower in survivors [(0.82 ± 1.70), (28.12 ± 51.17) µg/ml] than in nonsurvivors [(9.32 ± 12.04), (384.53 ± 597.93) µg/ml, respectively] (P < 0.01). The levels of leukocyte count, serum creatinine, aspartate aminotransferase (AST), and amylase were significantly higher in nonsurvivors than in survivors (P < 0.05, P < 0.01). In addition, metabolic acidosis was easier to appear in nonsurvivors. The multiple logistic regression analysis indicated that the paraquat concentration in plasma and urine, leukocyte count, creatinine and base excess were all related to survival.

CONCLUSIONThe higher paraquat concentration in plasma and urine, leucocytosis, renal dysfunction and metabolic acidosis are all important factors for the prognosis of paraquat intoxication.

Acidosis ; Adolescent ; Adult ; Aged ; Female ; Humans ; Kidney Diseases ; Leukocytosis ; Male ; Middle Aged ; Paraquat ; blood ; poisoning ; urine ; Prognosis ; Retrospective Studies ; Young Adult

Objective: To observe the intervention effect of auricular point sticking on dry eye in myopia patients after small-incision lenticule extraction (SMILE) surgery.Methods: This was a prospective randomized controlled study conducted among the myopia patients who received SMILE surgery at Eye and ENT Hospital of Fudan University. The post-SMILE operation patients who screened by the inclusion and exclusion criteria were randomized into a control group and a treatment group. Patients in the control group were given 0.1％ fluorometholone and 0.5％ carboxymethylcellulose sodium eye drops, while the treatment group was given additional unilateral auricular point sticking for 1 month. The patients were estimated using ocular surface disease index (OSDI), Schirmer tear test-1 (STT-1), tear film break-up time (TF-BUT), corneal fluorescein staining (CFS) score, corneal sensitivity (CS) and visual quality (VQ) at 1 d, 1 week and 1 month after surgery; the changes in anxiety and depression were also observed in the patients. Results: Compared with the first day after operation, CS in the nasal region was improved in the treatment group, and the VQ score increased in the control group patients at 1 week after operation (both P<0.05); at 1 month after operation, the TF-BUT increased, CFS score decreased, CS in the central and nasal regions increased (all P<0.05), and VQ score increased (P<0.01) in the treatment group, and the CS in the central, upper, lower and nasal regions were improved (all P<0.05), and VQ score increased (P<0.01) in the control group. The between-group comparison showed that the differences in the change of TF-BUT were statistically significant at 1 week and 1 month after surgery, respectively (both P<0.05). Conclusion: Auricular point sticking therapy can increase the TF-BUT and accelerate the repair of ocular surface function in post-SMILE patients.

OBJECTIVETo explore the acting mechanism of electrical field in electrical injury.

METHODSThirty-six New Zealand white rabbits were employed in the study and were randomly divided into 7 groups. There were 12 rabbits in group 1 and 4 in each group of other 6 groups. The animal model of nonthermal electrical injury previously replicated was employed in the study. Experiment with paralleled muscular fibers in electrical field was carried out in groups 2 approximately 4, while that of vertical muscular fibers in electrical field in groups 5-7. Anatomical examination was done to determine the index of deep burn injury (IDBI) in all groups of rabbits at 0, 2 and 24 postburn hour (PBH). Histological and ultrastructural examination, gamma picturing and isotope scanning with 99mTc were done in group 1 at 2 PBH.

RESULTSThere was no obvious skin injury in the white rabbits in group 1. Deep tissue necrosis was identified under the small electrode. Constant muscular spasm was observed in the inner side of the thigh. The muscles in paralleled electrical field suffered more severe injury than those in vertical one. Tissue injury was more severe in those areas with higher current density, less soft tissue, and also in the central area of the axis of the electric field. There were obvious changes in the perfusion and blood pool phases in these areas as observed with the aid of 99mTc. Light microscopic examination revealed swelling and necrosis of muscular fibers. Under electron microscopy, it was found that there were edema and dissolution with separation of lipid molecular layers of cell membrane, Shortened nucleus with partial dissolution of nuclear membrane, increased heparin granules within nucleus, swelling of mitochondria and endoplasmic reticulum, myofilament dissolution, expanded gap between myofilament and decreased number of heparin granules.

CONCLUSIONNon-thermal tissue injury in the electrical field, in terms of cell, ultrastructural and molecular levels, was induced and aggravated by all the factors constituting high voltage electrical field.

Animals ; Electric Injuries ; pathology ; Lower Extremity ; injuries ; Necrosis ; Rabbits ; Soft Tissue Injuries ; pathology