Background Research demonstrated that alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid GluR2 (AMPA-GluR2) is associated with amblyopia.It has been shown that levodopa and cytidine diphosphate choline can improve visual function of amblyopic children,but the mechanism is unclear.Objective This study was to explore the possible effects of levodopa and cytidine diphosphate choline on amblyopia.Methods Monocular deprivation (MD) animal models were created in 60 2-week-old SD rats by monolateral eyelid suturing and observed for 31 days and reared in natural light together with 15 other matched normal healthy SD rats.The models were randomly divided into the MD group,levodopa group,cytidine diphosphate choline group and normal saline control group,with 15 rats for each group.40 mg/kg of levodopa,80 mg/kg of cytidine diphosphate choline,I ml normal saline were given to the rats,respectively,for 28 consecutive days.Expressions of the AMPA-CluR2 protein and AMPA-CluR2 mRNA in the rat visual cortex were detected by immunohistochemistry,Western blot and real-time fluorescence quantitative PCR.Use of the animals followed the Regulations for the Administration of Affairs Concerning Experimental Animals by State Science and Technology Commission.Results The expression values of the AMPA-GluR2 protein (AMPA-GluR2/β-actin) and AMPA-GluR2 mRNA (2-△△Ct) were significantly lower in the MD group than those of the normal control group (protein:0.32 ± 0.02 vs.0.64 ± 0.05,t =13.287,P＜0.05 ;mRNA:0.30±0.01 vs.0.84±0.03,t=38.184,P＜0.05).Those in the levodopa group were significantly increased in comparison with the normal saline solution group (protein:0.59 ±0.04 vs.0.33 ±0.03,t =11.628,P＜0.05 ; mRNA:0.71±0.06 vs.0.33 ±0.02,t =13.435,P＜0.05).The expression values of the AMPA-GluR2 protein and AMPA-GluR2 mRNA were significantly increased in the cytidine diphosphate choline group compared with the normal saline solution group (protein:0.52 ± 0.04 vs.0.33 ± 0.03,t =8.497,P ＜ 0.05 ; mRNA:0.48± 0.04 vs.0.33 ± 0.02,t =7.500,P＜0.05).Conclusions AMPA-GluR2 is associated with the plasticity of visual development.Levodopa and cytidine diphosphate choline may improve visual function by down-regulating the expression of AMPA-GluR2 in the visual cortex.

Background The afferent signals of proprioceptor in extraocular muscles play an important role in controlling eye position and conjugate movement. Palisade ending in the extraocular muscles is the main source of proprioceptive information, and its abnormalities in structure and function may be associated with the occurrence of nystagmus. Objective This study was to observe the changes of palisade ending in the extraocular muscles of patients with congenital nystagmus ( CN) and discuss the probable mechanism. Methods Modified Kestenbaum procedure was performed on 10 patients with CN, and the extraocular muscle samples were collected during the operation. Normal extraocular muscle samples were obtained from the enucleated eyeballs after ocular wound. The ultrathin sections of extraocular muscles were prepared and double-staining by uranyl acetate and lead citrate. The morphological changes of the palisade ending of extraocular muscles were examined under the transmission electron microscopy. Written informed consent was obtained from each subject before surgery. Results The ultrastructure of palisade ending in the extraocular muscle of CN subjects showed the different degrees of alterations. The mild changes included the collapse and disconnection of external capsules and the nonhomogeneous electron-dense substracts. The degeneration and dissociation of myelin in nerve endings, swelling and vacuolation of mitochondria were also exhibited. Myeloid body was found in axon. In the severe patients,the necrosis of Schwann' s cells,dissolve of axon and disappear of capsules were seen. Conclusion The palisade ending of extraocular muscle in the patients with CN are obviously abnormal in comparison with normal one. These alterations are probably associated with the etiology and pathogenesis of CN.

· AIM: To observe the inhibitory effect of triamcinolone acetonide (TA) on the proliferation of monkey choroidretinal endothelial cells (RF/6A) in hypoxia or normal conditions.retinal endothelial cells of rhesus monkey (RF/6A). The effect of TA on the cellular activity was observed by MTT,the effect on cellular proliferation and apoptosis was detected by flow cytometry (FCM).cycle were reduced and the proportion of cells in G2-M phase was increased under the hypoxia condition .TA had a great effect on the cell cycle of choroid-retinal endothelial cells of rhesus monkey and it induced apoptosis of endothelial cells. It relatively increased the S-phase cells and reduced G2-M phase cells under both normal and hypoxia conditions, which indicates its role in blocking cell cycle from s-phase to G2-M phase and reducing mitosis.RF/6A cells while TA has the opposite effect in both normal and hypoxia conditions. TA can also induce apoptosis of endothelial cells.

8 ?g/ml were 7 strains in prophylactic treatment group and 3 strains in non-fluconazole prophylactic treatment group respectively.The two groups had significant difference (x~2=8.75,P

Objective:To investigate the expression of Bcl-2 and Bax in renal tissues of patients with hepatitis B virus- associated glomerulonephritis(HBV-GN).Methods:Twenty HBV-GN specimens with complete nephrology data and 10 normal renal specimens were randomly chosen for the present study.Cell apoptosis was detected by means of terminal deoxynucleotidyl transferase mediated d-UTP nick end labeling(TUNEL)and the apoptotic index was calculated;immunohistochemistry was used to detect the protein expression of Bax and Bcl-2.ResuLts:The apoptotic index in HBV-GN group was obviously higher than that of the control group;the apoptotic cells were mainly distributed in the proximal and distal renal tubules and the collecting duct epithelial cells,seldom seen in the glomerular cells.The expression of Bcl-2 in HBV-GN patients was predominately present in the renal tubular epithelia cells(positive in the plasma,membrane and nuclear);the expression of Bax was found in both glomerular cells and renal tubular cells,mainly in tubular epithelial cells,seldom seen in Bowman's capsule or glomerular mesangial region.Conclusion:Apoptosis in the kidney of HBV-GN patients mainly occurs in the renal tubular epithelial cells;expression of Bax and Bcl-2 is mainly in the renal tubular epithelial cells,suggesting that the injury of tubular interstitial damage may be one of the important factors for the development of HBV-GN.

AIMTo investigate the effects of curcumin on the behavior of chronic constrictive injury (CCI) rats and the p-ERK, p-CREB, c-fos expression in dorsal root ganglion.

METHODS108 male SD rats were randomly divided into 6 groups: (1) Control group (treated with CCI); (2) Sham operation group; (3) Solvent contrast group; (4) Curcumin treated group(Cur 30, Cur 100, Cur 300), treated with CCI, intraperitoneal injected with curcumin 30 mg x kg(-1) x d(-1), 100 mg x kg(-1) x d(-1), 300 mg x kg(-1) x d(-1) for 14 days after operation respectively. Thermal withdrawal latency (TWL) and mechanical withdrawal threshold (MWT) of rats were determined, respectively. Rats were killed on the 3th, 7h, 14th day after operation. The expression of p-ERK, p-CREB, c-fos in dorsal root ganglion were assessed by immunohistochemical analysis.

RESULTSIn Con group, the MWT and TWL declined gradually after operation. On the 3rd day, the rats represented the severest mechanical and thermal hyperalgesia(MWT was 15.3 +/- 3.0 g, TWL was 4.6 +/- 1.0 s). The expression of p-ERK, p-CREB, c-fos neurons were markedly increased in dorsal root ganglion. In Cur group, the MWT and TWL were also declined gradually, which were higher than Con group. On the 3rd day, the rats represented the severest mechanical and thermal hyperalgesia (MWT was 22.6 +/- 4.0 g, TWL was (5.6 +/- 1.1l)s in Cur 100 group), the expression of p-ERK, p-CREB, c-fos in dorsal root ganglion were lower than control group at each timepoint in each group.

CONCLUSIONCurcumin could attenuate the activation of p-ERK, p-CREB, c-fos in dorsal root ganglion to ameliorate the CCI-induced neuropathic pain.

Animals ; CREB-Binding Protein ; metabolism ; Curcumin ; pharmacology ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Ganglia, Spinal ; drug effects ; metabolism ; Male ; Neuralgia ; metabolism ; Proto-Oncogene Proteins c-fos ; metabolism ; Rats ; Rats, Sprague-Dawley

As the dilution procedure was applied, a simple, rapid and cost-effective high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for determination of aflatoxin B1, B2, G1, and G2 was successfully by performed in a total 83 samples of 10 traditional Chinese medicines (TCMs), which were collected from 5 different hospital pharmacies and 5 different medical stores in Guangzhou city. Matrix effects of these 10 TCMs were ranged from 80.23% to 115.5% in low, intermediate and high concentration levels, indicating that the negative effect was overcome in this study. Meanwhile, the analysis method was proved to be stable and reliable during the whole analysis using Semen Armeniacae Amarum spiked 3 concentration levels of standard solution as quality control samples and the RSD < 6.6% was obtained. The contamination levels of 83 investigated samples were 13.89% and 17.02% in hospital pharmacies and medical stores, respectively. The result was presented to provide relevant reference and supplement to those researchers in TCMs analysis and screening.
Aflatoxin B1 ; analysis ; Aflatoxins ; analysis ; Chromatography, High Pressure Liquid ; methods ; Drug Contamination ; Medicine, Chinese Traditional ; Quality Control ; Tandem Mass Spectrometry ; methods

BACKGROUNDGlucocorticoid is speculated to be able to have Aspergillus fumigatus (A. fumigatus) being more susceptible to reactive oxygen species (ROS) by inhibiting Afyap1, the transcription factor activating protein-1 (AP-1) homologue in A. fumigatus, which may provide a clue to expand the clinical use of glucocorticoid in patients with fungal infections. In this study, we used dexamethasone to determine the direct effect on oxidative killing susceptibility of A. fumigatus in vitro, as well as the expression level of Afyap1 gene and its target genes (catalase and superoxide dismutase (SOD) genes).

METHODSA. fumigatus spores were treated with different concentrations (0, 0.02, 0.2 mg/ml) of glucocorticoids and assigned to four groups (A: 0.5 hour, B: 2 hours, C: 7 hours, D: 16 hours) according to the time of treatment. The H2O2 oxidative killing assay was done, using the standard method-spot test, in each group of A. fumigatus. We measured the oxidative killing susceptibility as well as the expression level of the gene Afyap1, CATA, SOD1 and SOD2 in A. fumigatus at each group. The antifungal susceptibility to itraconazole and amphotericin B in each group of A. fumigatus was also measured with M38-A2 method.

RESULTSThe oxidative killing susceptibility of A. fumigatus was increased, consistent with the reduction of Afyap1, CATA, SOD1 and SOD2 gene expression level after being treated with dexamethasone for 0.5 hours. However, these observations were disappeared along with being treated for longer time. The antifungal susceptibility to itraconazole and amphotericin B in the A. fumigatus strains treated with dexamethasone indicated no change, compared with those without dexamethasone treatment.

CONCLUSIONDexamethasone can have A. fumigatus being more susceptible to ROS when treated for shorter period (0.5 to 2 hours) via the reduction of Afyap1 gene expression as well as the down-stream enzyme-coding gene expression.

Aspergillus fumigatus ; drug effects ; genetics ; metabolism ; Dexamethasone ; pharmacology ; Fungal Proteins ; genetics ; metabolism ; Hydrogen Peroxide ; pharmacology

OBJECTIVECleidocranial dysplasia (CCD) is a dominantly inherited skeletal dysplasia caused by mutations in the osteoblast-specific transcription factor-encoding gene, core binding factor α1 (CBFA1). Over 90 mutations in CBFA1 gene have been published to date in 500 independent cases of CCD, including missense mutations, deletions, insertions, frameshift, and splice mutations. However, mutational screening of the CBFA1 gene is still far from saturation, and more novel mutations will be identified to enrich the insights into the molecular basis for the pathogenesis of CCD. The aim of this study was to explore the clinical and image features and detect the mutations of CBFA1 gene in two CCD families.

METHODIn this study, the clinical features were investigated in two CCD families, radiological and CT examinations regarding osseous malformation were carried out over the entire body of these patients with CCD. Blood (2 ml) was drawn from all affected individuals, unaffected family members and one hundred unrelated normal controls, Genomic DNA was extracted from whole blood with PureGene DNA extraction kit and PCR was performed with eight pairs of PCR primers for exons 0 to 7 of the CBFA1 gene. The mutations of CBFA1 gene were screened in these two CCD families.

RESULT(1) The clinical features of patients with CCD include delayed closure of fontanelles, frontal bossing, dysplasia of clavicles, late tooth eruption, and other skeletal anomalies. X-ray and CT examination showed the bulging calvarium, patent fontanelles, wide cranial sutures, multiple Wormian bones, dental dysplasia or aplasia of clavicles. (2) Two mutations were identified, one is novel missense mutation (c.1259C > T[p.T420I]) in CBFA1 gene exon 7, other (c.577C > T[p.R193X]) was reported in Chinese cases with CCD for the first time.

CONCLUSION(1) The clinical and image features of patients in two CCD families include delayed closure of fontanelles, frontal bossing, dysplasia of clavicles, late tooth eruption, and other skeletal anomalies. (2) The T420I and R193X mutations of CBFA1 were reported, expanding the spectrum of CBFA1 mutations causing CCD.

Child ; Child, Preschool ; Cleidocranial Dysplasia ; genetics ; pathology ; Core Binding Factor Alpha 1 Subunit ; genetics ; DNA Mutational Analysis ; Exons ; Female ; Humans ; Male ; Mutation ; Pedigree ; Phenotype

AIMTo investigate the stage-specific localization of metastasis-associated protein 1 (MTA1) during spermatogenesis in adult human and mouse testis.

METHODSThe immunolocalization of MTA1 was studied by immunohistochemistry and Western blot analysis. The distribution pattern of MTA1 in mouse testis was confirmed by using quantitative analysis of purified spermatogenic cells.

RESULTSThe specificity of polyclonal antibody was confirmed by Western blot analysis. MTA1 was found expressed in the nucleus of germ cells, except elongate spermatids, and in the cytoplasm of Sertoli cells; Leydig cells did not show any specific reactivity. MTA1 possessed different distribution patterns in the two species: in humans, the most intensive staining was found in the nucleus of round spermatids and of primary spermatocytes while in mice, the most intense MTA1 staining was in the nucleus of leptotene, zygotene and pachytene spermatocytes. In both species the staining exhibited a cyclic pattern.

CONCLUSIONThe present communication initially provides new evidence for the potential role of MTA1 in mature testis. In addition, its distinctive expression in germ cells suggests a regulatory role of the peptide during spermatogenesis.

Adult ; Animals ; Animals, Outbred Strains ; Blotting, Western ; Histone Deacetylases ; metabolism ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Repressor Proteins ; metabolism ; Sexual Maturation ; physiology ; Species Specificity ; Spermatogenesis ; physiology ; Testis ; cytology ; metabolism ; Transcription Factors ; metabolism