Amblyopia greatly affects the physical and mental development of children. Acupuncture is effective for amblyopia, though its mechanism remains unclear. This article summarized the mechanism of acupuncture treatment of amblyopia from the perspectives of morphology of neurons in visual cortex, visual electrophysiology, and molecular biology, etc. It was found that acupuncture may treat amblyopia through repairing the morphological and ultrastructural damages of neurons in visual cortex, promoting the electrical activities in visual pathway and visual cortical neurons, and modulating the synthesis and expression levels of factors involved in visual system. Nevertheless, further studies are required to unveil the mechanism of acupuncture treatment of amblyopia.

Objective To investigate histopathological changes and expression of integrin ?1 of sternomastoid muscle,and probe the mechanism and significance during disease process in congenital muscular torticollis(CMT).Methods Histopathological changes of sternomastoid muscle section stained with HE and Gomori silver staining were observed and the expression of integrein ?1 with immunohistochemistry was detected,and the expressive quantity and distribution with image analysis system was quantitive analyzed.Results 1.With light microscopy observation,the results showed that the fibrous degeneration of sternomastoid muscle could be summed up 2 kinds: A category displayed the myocytes atrophyed,and there were lots of connective tissue hyperplasy around myocytes,and the direction of fibrous arrangement was disordered,meanwhile there were lots of vessels and nervers hyperplasy,and eventually the myocytes shrank back and disappeared.B category displayed that the structure of cross striation or sarcomere disappeared or changed,and myocytes could maintained the outline and the sarcolemma were integrated,and then fibrous pathological changes of myocyte took place,and there were lots of fibroblast-like that had much more enations between fiber bundles.With Gomori silver staining,the major changes of fibrotic sternomastoid muscle showed that there were lots of collagenous fibers hyperplasy.The arrangements of collagenous fibers were disordered in A category and were well-arranged in B ca-tegory.2.With immunohistochemistry,the results showed the expression of integrin ?1 was weak positive in normal control group(125.7?5.167).In diseased groups,the results showed 3 different extents:the expression of integrin ?1 displayed stronger positive in A category myocytes(30.15?6.543),and the level of expression was significantly different from normal controls(P0.05).Conclusions The fibrous pathological changes of sternomastoid muscle are a complicated and gradually process,which may has different mode,and ingetrin ?1 may participated the process of pathological changes.

Objective To observe and analyze the performance of forensic science in the cases of suxamethonium chloride poisoning,and to improve the identification of suxamethonium chloride poisoning.Methods Fifty-four cases of suxamethonium chloride poisoning were collected.The rules of determination of suxamethonium chloride poisoning were observed by the retrospective analysis of pathological and toxicological changes as well as case features.Results The pathological features of suxamethonium chloride poisoning were similar to the general changes of sudden death,which mainly included acute pulmonary congestion and edema,and partly showed myocardial disarray and fracture.Suxamethonium chloride could be detected in the heart blood of all cases and in skin tissue of part cases.Conclusion Suxamethonium chloride poisoning has the characteristics with fast death and covert means,which are difficult to rescue and easily miss inspection.For the cases of sudden death or suspicious death,determination of suxamethonium chloride should be taken as a routine detection index to prevent missing inspection.

OBJECTIVETo explore the binding domain of hydroxypyruvate isomerase homologues (HYI) in the interaction with protein P311 in hypertrophic scar fibroblasts (Fb).

METHODS(1) P 311 was amplified by PCR using plasmid pMD18-T-P 311 as template. The total RNA of hypertrophic scar Fb was extracted by Trizol to amplify HYI with RT-PCR. Recombinant vectors pGADT7-P 311 and pGBKT7-HYI were constructed by double-enzyme digestion, and they were verified by PCR and sequencing. The secondary structure of protein HYI was analyzed with software Prot Seale and HNN. Fragments of HYI-1 (1-447 bp), HYI-2 (247-447 bp), HYI-3 (1-279 bp), and HYI-4 (247-654 bp) were amplified based on the result of software analysis. And then the recombinant vectors pGBKT7-HYI-1, 2, 3, and 4 were constructed by double-enzyme digestion and verified by PCR and sequencing. (2) AH109 yeast cells were transformed into competent cells by lithium acetate method and divided into 7 groups roughly in the same amount, including HYI full length, HYI-1, HYI-2, HYI-3, and HYI-4 hybrid groups, positive control group, and negative control group. Cells in the first five groups were respectively transformed with recombinant vector pGBKT7-HYI full length, pGBKT7-HYI-1, pGBKT7-HYI-2, pGBKT7-HYI-3, pGBKT7-HYI-4 and recombinant vector pGADT7-P 311, and that in the rest two groups were transformed with recombinant vectors pGBKT7-53 and pGADT7-RecT, pGADT7-RecT and pGBKT7-Lam by polyethyleneglycol/lithium acetate method. Immediately after transformation, a part of the transformed cells in each group was spread onto the medium lacking leucine, tryptophan, adenine, and histidine (briefly called four-factor lacking medium), and another portion of the cells was spread onto the medium lacking leucine and tryptophan (briefly called two-factor lacking medium). After 3 to 6 days' culture, the growth of yeast was observed, and the expression of β-galactosidase of yeast was detected by color reaction with 5-bromo-4-chloro-indolyl-β-D-galactopyranoside.

RESULTS(1) Cloned P 311 and the reported P 311 (GenBank ID hsu36189) had the same sequence. The A base at 496 bp in reported HYI (GenBank ID AY775560) was replaced by G base as found in cloned HYI. It was verified that the insert segment of each recombinant vector was correct. (2) Among those 216 amino acids which composed the protein HYI, 101 amino acids might form α helices, 90 amino acids might form random coils, 25 amino acids might form extended-chains as revealed in the simulated structure analysis by computer. (3) Cloned segments HYI-1, 2, 3, 4 showed expected lengths. It was verified that the insert segment of each recombinant vector was correct. (4) Except for strains in negative control group which did not show growth on four-factor lacking medium, all strains in other groups grew on both kinds of media, and growth of colonies was less in HYI-2 (with the fewest number of α helices) and HYI-3 hybrid groups. (5) Positive expression of β-galactosidase was observed in strains of all groups growing on four-factor lacking medium except for the HYI-2 hybrid group. No expression of β-galactosidase was observed in strains of negative control group which grew on two-factor lacking medium.

CONCLUSIONSProtein HYI may closely bind with protein P311 by α helix, which plays an important role in fibroblast-to-myofibroblast transdifferentiation in hypertrophic scar.

Aldose-Ketose Isomerases ; genetics ; Cicatrix, Hypertrophic ; enzymology ; genetics ; Cloning, Molecular ; Fibroblasts ; enzymology ; Genetic Vectors ; Humans ; Molecular Sequence Data ; Nerve Tissue Proteins ; metabolism ; Oncogene Proteins ; metabolism ; Plasmids ; Protein Binding ; Protein Interaction Domains and Motifs

OBJECTIVETo investigate the dose and time-dependent effects of lipopolysaccharide (LPS) on cytoskeletal F-acitn and G-actin reorganizations by visualizing their distribution and measuring their contents in human umbilical vein endothelial cell line ECV-304.

METHODSF-actin was labeled with rhodamine-phalloidin and G-actin with deoxyribonuclease I (DNase I)conjugated with fluorescein isothiocyanate (FITC). Contents of cytoskeletal proteins were obtained by flow cytometry.

RESULTSF-actin was mainly distributed peripherally in endothelial cells under normal conditions. LPS stimulation caused the formation of stress fibers and filopodia. G-actin was normally seen in perinuclear and nuclear areas in control ECV-304 cells. Under LPS stimulation, G-actin dots appeared in the cytoplasmic region. The actin disorganization was accompanied by the time- and dose- dependent decrease in F-actin pool and increase in G-actin pool.

CONCLUSIONSLPS can induce characteristic morphological alterations of actin cytoskeleton and formation of intercellular gap in endothelial cells, accompanied by changes in F-actin and G-actin pools.

Actins ; drug effects ; Analysis of Variance ; Cells, Cultured ; Deoxyribonuclease I ; Dose-Response Relationship, Drug ; Endothelial Cells ; chemistry ; Escherichia coli ; Fluorescein-5-isothiocyanate ; Fluorescent Dyes ; Humans ; Lipopolysaccharides ; pharmacology ; Phalloidine ; Rhodamines ; Umbilical Veins ; cytology

OBJECTIVETo investigate the changes in endothelial cytoskeletal reorganization and the role of Rho in the signal transduction pathway.

METHODSECV304 cells were cultured and randomly divided into following groups: i.e. sham (with normal rat serum treatment), burn (with burn rat serum treatment), Y (with 30 micromol/L Rho kinase inhibitor Y-27632 treatment), burn plus Y (pretreatment of cells with burn serum before treated with 30 micromol/L Y-27632), Y plus burn (pretreatment of cells with Y-27632 for 1 hour before treated with burn serum), LPA (with normal rat serum and 13 micromol/L LPA), and LPA plus Y (pretreatment of cells with LPA before treated with Y-27632) groups. The indices were examined at 6, 7 and 8 posttreatment hours (PTH) in all groups except in Y group. The endothelial morphology was observed with HE staining. Endothelial cytoskeleton was observed by dual-fluorescence labeling of filamenta (F) with Rhodamine-phalloidin and monomer (G) with oregon green labeled DNAase. The actin content in the cells in all groups was measured with flow cytometry.

RESULTSIn sham and control group, the cells were in fusiform or polygonal shape, with satisfactory growth; filamentous actin (F-actin) was mainly distributed in the peripheral site of the cytoplasm and formed peripheral filamental band. The cells became confluent to form a single layer with reticular structure. Globular actin (G-actin) was concentrated in the nucleus and per nucleus. In burn group, after 6 hours of burn serum treatment, the ability of cells to adhere to vessel wall was weakened, and a striking reorganization of the actin cytoskeleton and the formation of the stress fibers were found. Furthermore, the fluorescent intensity of the peripheral filament bands was weakened, and dispersed actin monomers were seen in the cytoplasm. This reaction was enhanced along with elapse of stimulation time. In burn plus Y or Y plus burn group, the cells grew and adhered well to the wall of culture vessel. The distribution of the filamentous actin was the same as the sham group, while the stress fiber decreased in amount obviously. The structure of globular actin was condensed with little G-actin in the cytoplasm. The changes in actin cytoskeleton in LPA group was similar to that in burn group. The effects of LPA on actin reorganization could also be reversed by Y-27632. The content of F-actin in burn group at 6 PTH (0.63 +/- 0.07) was lower than that in sham group (0.75 +/- 0.08), while the content of G-actin in burn group (1.28 +/- 0.27) was higher than that in sham group (1.16 +/- 0.16, P > 0.05).

CONCLUSIONBurn serum induces vascular endothelial actin cytoskeleton reorganization in endothelial cells via the Rho-dependent signal pathway. Similar to the effect of LPA, this effect could be reversed by Y-27632.

Actins ; metabolism ; Amides ; pharmacology ; Animals ; Burns ; blood ; metabolism ; Cells, Cultured ; Cytoskeleton ; metabolism ; Endothelial Cells ; metabolism ; Endothelium, Vascular ; Humans ; Male ; Pyridines ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Serum ; metabolism ; Signal Transduction ; rho-Associated Kinases ; antagonists & inhibitors ; metabolism

OBJECTIVETo compare the acute pulmonary toxicities of nanosized and microsized silicon dioxide particles.

METHODSAll 125 healthy male Wistar rats were divided into 25 groups randomly according to the weight. Experimental animals were exposed to microsized SiO2 at the doses of 100 mg/m3 (group A) and 300 mg/m3 (group B), and to the nanosized SiO2 at the same dose levels (group A' and B') by inhalation for 2 hours. Compositions in bronchoalveolar lavage fluids (BALF) and contents of hydroxyproline in blood sera and lung tissues were detected and then compared at 6, 12, 24, 48, 72 hours after administration.

RESULTSThe total cellular score (TCS) in BALF of group A'[(55.00 +/- 8.30) x 10(4)/ ml] and B'[(52.50 +/- 9.02) x 10(4)/ml] at 6 hours were significantly higher than those in control groups [(34.88 +/- 12.53) x 10(4)/ml]; TCS in BALF of group A' [(55.00 +/- 8.30) x 10(4)/ml]at 6 hours and group A' [(39.75 +/- 12.08) x 10(4)/ml] at 24 hours were significantly higher than those in isodose group of microsized SiO2 [(32.38 +/- 13.07) x 10(4)/ml, (24.13 +/- 10.97) x 10(4)/ml) ]; total protein (TPr) in BALF of group A' [(0.34 +/- 0.09)g/L] and B' [(0.38 +/- 0.16) g/L] at 48 hours were significantly higher than those in isodose group of microsized SiO2 [(0.20 +/- 0.07) g/L, (0.21 +/- 0.05) g/L]. Lactate dehydrogenase (LDH) in BALF of group A' [(1.66 +/- 0.22) x 10(3) U/L] at 72 hours were significantly higher than those in isodose group of microsized SiO2 [(1.38 +/- 0.17) x 10(3) U/L]. Alkaline phosphatase (AKP) in BALF of group B' [(5.14 +/- 1.47) U/100 ml] at 6 hours and group B' [(5.86 +/- 2.41) U/100 ml] at 24 hours were significantly higher than those in isodose group of microsized SiO2 [(3.64 +/- 0.36) U/100 ml, (3.30 +/- 2.19) U/100 ml]. Hydroxyproline (HyP) in tissues of lung of group A' [(0.532 +/- 0.053) microg/mg, (0.484 +/- 0.046) microg/mg, (0.591 +/- 0.096) microg/mg, (0.551 +/- 0.084) microg/mg] at 6, 12, 48, 72 hours and group B' [(0.508 +/- 0.081) microg/mg, (0.565 +/- 0.053) microg/mg ] at 12, 72 hours were significantly higher than those in isodose group of microsized SiO2 [(0.345 +/- 0.074) microg/mg, (0.368 +/- 0.095) microg/mg, (0.431 +/- 0.036) microg/mg, (0.399 +/- 0.080) microg/mg, (0.396 +/- 0.039) microg/mg, (0.465 +/- 0.062) microg/mg].

CONCLUSIONNanosized and microsized SiO2 should have some differences on acute pulmonary toxicities in our experiment condition.

Animals ; Bronchoalveolar Lavage Fluid ; Dust ; Lung ; drug effects ; Male ; Nanoparticles ; Particle Size ; Rats ; Rats, Wistar ; Silicon Dioxide ; toxicity ; Toxicity Tests, Acute

OBJECTIVETo analyze the motor performance status in students of Han nationality in fifteen provinces in china.

METHODSTotally, 161 804 students of Han nationality aged from 7 to 18 years old were involved in the Chinese Surveillance on Students' Physical Fitness and Health in 2004. Motor abilities were accessed with the aid of gripping power, 50 m dash, standing long jump, and 1-min sit-ups. Based on general statistical description, principal component analysis and linear regression, the development characters of students' motor performance were explored.

RESULTSThis research showed some characters similar to those of last ones: motor capability was improved in correlation with age; boys did better than girls, the difference between 18 year-old rural boys and rural girls was 15.3 kg, -2.0 s, 58.6 cm, 8.7/min; the urban students performed better than the rural ones, the difference between 15 year-old urban boys and rural boys was 0.9 kg, -0.2 s, 3.5 cm, 3.5/min. The first principal component might represent the 4 tests greatly. Regression analysis on principal component quantitatively interpreted the influence of factors such as age, sex and area.

CONCLUSIONSThe general principles of exercise quality development of students are still in work. Principal component analysis should be adequate and convenient in motor performance analysis.

Adolescent ; Body Constitution ; Body Mass Index ; Child ; China ; Female ; Humans ; Male ; Physical Education and Training ; Physical Fitness ; Population Surveillance ; methods ; Rural Population ; statistics & numerical data ; Students ; statistics & numerical data ; Urban Population ; statistics & numerical data

To investigate and compare the expression of intercellular adhesion molecule-1 (ICAM-1) in different organs of the mice with endotoxic shock induced by lipopolysaccharide (LPS), protein and mRNA of ICAM-1 were measured by Western blotting and RT-PCR respectively in different organs of BALB/c mice administered intraperitoneally with 5 mg/kg LPS. The results showed that the constitutive expression of ICAM-1 protein and mRNA was the greatest in the lungs, followed by the spleen, kidney and intestine. After LPS stimulation, the upregulation of ICAM-1 was still greatest in the lungs, followed by the liver, spleen, heart, kidney and intestine. Compared with the normal mice, the expression of ICAM-1 protein in endotoxic shocked mice increased by 4.5-fold in the lungs, 3.0-fold in the kidney, 1.5-fold in the spleen; the expression in the liver and heart was negative under normal condition and changed into positive during endotoxic shock; but ICAM-1 expression in the intestine did not change significantly. The expression of ICAM-1 mRNA also increased consistently. These data highlight that LPS can up-regulate ICAM-1 protein and mRNA expression in different tissues of the mice with endotoxic shock. The difference in ICAM-1 expression among the organs may lead to different sensitivity of organ damage in endotoxic shock. This suggests that inhibition of ICAM-1 expression may be a useful principle for prevention and treatment of endotoxic shock.
Animals ; Intercellular Adhesion Molecule-1 ; biosynthesis ; Kidney ; metabolism ; Lipopolysaccharides ; Lung ; metabolism ; Male ; Mice ; Mice, Inbred BALB C ; RNA, Messenger ; biosynthesis ; Shock, Septic ; chemically induced ; metabolism

AIM:To investigate the angiogenic effect and mechanisms of astragaloside IV(AS-IV)in rats with myocardial infarction via protein kinase D 1(PKD1)-histone deacetylase 5(HDAC5)-vascular endothelial growth factor (VEGF)signaling pathway.METHODS:The classic model of myocardial infarction by ligation of the left anterior de-scending coronary artery was replicated,and the rats were randomly divided into model group,AS-IV group,and AS-IV+CID755673(PKD1 inhibitor)group.The sham operation control group and DMSO control group were also set up.All the rats were given intravenous injection via caudal vein.The rats were sacrificed 4 weeks later,and segmental heart samples were used for HE staining and Masson staining.The expression of PKD1,HDAC5 and VEGF was analyzed by immunohis-tochemistry,RT-PCR and and Western blot.RESULTS:Compared with sham operation group and DMSO group,the myo-cardium in model group showed disordered arrangement, accompanied with necrotic myocardial cells and obvious fibrosis tissue.After treatment with AS-IV,the morphological changes of myocardium were obviously improved,and the number of new blood vessels increased significantly.However,after treatment with AS-IV+CID755673,the myocardial tissues of the rats became disordered again,with increased necrotic cells and some closed vessels.The mRNA and protein expression of PKD1,HDAC5 and VEGF in myocardial tissue in model group was significantly lower than that in sham operation and DMSO groups(P<0.05).The expression in AS-IV group was significantly higher than that in model group(P<0.01), while that in AS-IV+CID755673 group was significantly lower than that in AS-IV group(P<0.05).CONCLUSION:AS-IV promotes the angiogenesis of myocardial tissues in the rats after myocardial infarction partly by regulating the PKD 1-HDAC5-VEGF signaling pathway.