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.

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

OBJECTIVETo study the expression of histone acetyltransferases (HATs) and histone deacetylases (HDACs) in children with tetralogy of Fallot (TOF), and to investigate the role of histone acetylation and acetylation-related enzymes in the pathogenesis of TOF.

METHODSMyocardial tissue samples in the TOF group were obtained from 46 children with TOF who underwent radical operation, and myocardial tissue samples in the control group were obtained from 16 children who suffered accidental deaths and had no cardiac anomalies as shown by autopsy. The acetylation of H3K9, H3K18 and H3K27 was evaluated by immunohistochemistry. The mRNA expression of HATs and HDACs in the myocardium was measured by real-time PCR. The correlation between mRNA expression of HATs and HDACs and histone acetylation was analyzed.

RESULTSCompared with the control group, the TOF group showed significantly increased acetylation of H3K9 (P=0.0165) and significantly decreased acetylation of H3K18 (P=0.0048) and H3K27 (P=0.0084). As to 4 HATs and 6 HDACs, the mRNA expression of EP300 and CBP was significantly higher in the TOF group than in the control group (P=0.025; P=0.017), and there was no significant difference in the mRNA expression of other HATs and HDACs between the two groups. The correlation analysis revealed a positive correlation between H3K9 acetylation and mRNA expression of EP300 (r=0.71, P<0.01) and CBP (r=0.72, P<0.01).

CONCLUSIONSUpregulated mRNA expression of EP300 and CBP may be associated with increased H3K9 acetylation, suggesting that EP300 and CBP might affect cardiac development by regulating H3K9 acetylation.

Acetylation ; E1A-Associated p300 Protein ; genetics ; Female ; Histone Acetyltransferases ; genetics ; Histone Deacetylases ; genetics ; Histones ; metabolism ; Humans ; Infant ; Male ; Myocardium ; metabolism ; Peptide Fragments ; genetics ; RNA, Messenger ; analysis ; Sialoglycoproteins ; genetics ; Tetralogy of Fallot ; metabolism

AIMTo investigate the stage-specific localization of transforming growth factor (TGF) beta1 and beta3 during spermatogenesis in adult human testis.

METHODSThe localization of TGFbeta1 and beta3 was investigated by immunohistochemical staining method employing specific polyclonal antibodies.

RESULTSBoth TGFbeta1 and beta3 and their receptors were preponderant in the Leydig cells. TGFbeta1 could not be detected in the seminiferous tubules. TGFbeta3 and TGFbeta-Receptor (R) I were mainly seen in the elongated spermatids, while TGFbeta-RII in the pachytene spermatocytes and weak in the spermatogonia, spermatids and Sertoli cells. Only TGFbeta-RII was detected in the Sertoli cells. TGFbeta3, TGFbeta-RI and TGFbeta-RII showed a staining pattern dependent upon the stages of the seminiferous epithelium cycle.

CONCLUSIONTGFbeta isoforms and their receptors are present in the somatic and germ cells of the adult human testis, suggesting their involvement in the regulation of spermatogenesis.

Adult ; Humans ; Immunohistochemistry ; Leydig Cells ; metabolism ; Ligands ; Male ; Middle Aged ; Orchiectomy ; Prostatic Neoplasms ; pathology ; Receptors, Transforming Growth Factor beta ; metabolism ; Seminiferous Epithelium ; cytology ; metabolism ; Spermatids ; metabolism ; Spermatogenesis ; physiology ; Testis ; metabolism ; physiology ; Transforming Growth Factor beta ; metabolism ; Transforming Growth Factor beta1 ; Transforming Growth Factor beta3

AIMTo study the expression and regulation of Smad1, Smad2 and Smad4 proteins (intracellular signaling molecules of transforming growth factor-b family) in rat testis during postnatal development.

METHODSThe whole testes were collected from SD rats aged 3, 7, 14, 28 and 90 (adult) days. The cellular localization and developmental changes were examined by immunohistochemistry ABC method with the glucose oxidase-DAB-nickel enhancement technique. Quantitative analysis of the immunostaining was made by the image analysis system. The Smads proteins coexistence in the adult rat testis was tested by the double immune staining for CD14-Smad4 and Smad2-Smad4. The protein expression of Smad during rat testicular development was examined by means of Western blots.

RESULTSSmad1, Smad2 and Smad4 were present throughout testicular development. The immunostaining of Smad1 and Smad2 were present in spermatogenic cells. A positive immunoreactivity was located at the cytoplasm, but the nucleus was negative. Smad1 was immunolocalized at the d14, d28 and adult testes, while Smad2, at the d7, d14, d28 and adult testis. There was positive immunoreaction in the Sertoli cells and Leydig cells as well. The immunolocalization of Smad4 was exclusively at the cytoplasm of Leydig cells and the nuclei were negative throughout the testicular development. No expression was detected in the germ cells. The results of image and statistical analysis showed that generally the expression of Smad1, Smad2 and Smad4 in the testis tended to increase gradually with the growth of the rat.

CONCLUSIONThe present data provide direct evidences for the molecular mechanism of TGF-bgr action in rat testes during postnatal development and spermatogenesis.

Animals ; Blotting, Western ; DNA-Binding Proteins ; analysis ; biosynthesis ; Immunohistochemistry ; Male ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; physiology ; Smad Proteins ; Smad1 Protein ; Smad2 Protein ; Smad4 Protein ; Testis ; chemistry ; growth & development ; physiology ; Trans-Activators ; analysis ; biosynthesis

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

OBJECTIVETo examine the effect of docosahexaenoic acid (DHA) deficiency in brain on spatial learning and memory in rats.

METHODSSprague Dawley rats were fed with an n-3 fatty acid deficient diet for two generations to induce DHA depletion in brain. DHA in seven brain regions was analyzed using the gas-liquid chromatography. Morris water maze (MWM) was employed as an assessing index of spatial learning and memory in the n-3 fatty acid deficient adult rats of second generation.

RESULTSFeeding an n-3 deficient diet for two generations depleted DHA differently by 39%-63% in the seven brain regions including cerebellum, medulla, hypothalamus, striatum, hippocampus, cortex and midbrain. The MWM test showed that the n-3 deficient rats took a longer time and swam a longer distance to find the escape platform than the n-3 Adq group.

CONCLUSIONThe spatial learning and memory in adult rats are partially impaired by brain DHA depletion.

Animals ; Brain ; metabolism ; Docosahexaenoic Acids ; metabolism ; Maze Learning ; physiology ; Memory ; physiology ; Rats ; Rats, Sprague-Dawley

In this study, an analytical method was developed and used to quantify simultaneously protocatechuic acid, neochlorogenic acid, cryptochlorogenic acid and 1, 3-dicaffeoylquinic acid--four bioactive compounds contained in Fructus Xanthii using UPLC. The contents of four phenolic components of 28 batches of samples collected from different product areas and markets were determined and compared by means of this established method. The mobile phase was composed of methanol and water containing 0.1% phosphoric acid. Chromatography was monitored at dual-wavelengths--220 and 327 nm. Flow rate was 0.4 mL x min(-1) and column temperature was 35 degrees C. The correlation coefficient between concentration and chromatographic peak area of protocatechuic acid, neochlorogenic acid, cryptochlorogenic acid and 1, 3-dicaffeoylquinic acid was over 0.9999 in the range of 0.3570-35.70, 2.500-250.0, 1.060-106.1, 1.010-101.0 microg x mL(-1), respectively. The average recoveries of the four compounds were 97.68%, 99.55%, 97.92% and 100.4%, respectively. In conclusion, the established method can rapidly attain an accurate and reproducible result used to control the quality of Fructus Xanthii.
Chlorogenic Acid ; analysis ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; analysis ; Fruit ; chemistry ; Hydroxybenzoates ; analysis ; Plants, Medicinal ; chemistry ; Quality Control ; Quinic Acid ; analogs & derivatives ; analysis ; Reproducibility of Results ; Xanthium ; chemistry

Objective To evaluate the effect of traditional Chinese medicine nursing technology among patients with ulcerative colitis.Methods Totally 170 patients with ulcerative colitis were recruited.All patients received routine treatment and received transitional care in inpatient department including diet guidance,illness response and anal guidance on contraction exercise,etc.What's more,the observation group implemented traditional Chinese medicine nursing techniques,including auricular pressure paste,acupuncture point application,acupressure,moxibustion,etc.After 7 and 14 days,we compared the patients' disease activity,sleep quality by Grading scale for Traditional Chinese Medicine symptoms and signs of ulcerative colitis,improved Mayo question,Pittsburgh Quality Sleep Index(PQSI).Results On the 7th and 14th day after intervention,the scores of symptoms and signs in the observation group were (9.73 ± 3.49),(4.54 ± 2.75) points,lower than those in the control group (10.72 ± 4.00),(8.22 ±3.06) points,and the difference was statistically significant (t =1.973,7.878,P =0.042,0.000).On the 14th day after intervention,the Mayo score of observation group was (3.33 ± 1.82) points,lower than that of control group (4.56 ± 1.77) points.The difference was statistically significant (t =4.278,P =0.000).The PSQI score of observation group was (5.67 ± 2.87) points,lower than that of control group (8.53 ± 3.13)points.The difference was statistically significant (t =5.934,P =0.000).Conclusions Traditional Chinese medicine transitional care could effectively improve the symptom of disease and sleep quality among patients with ulcerative colitis.

OBJECTIVETo understand HBV serotypes and genotypes epidemiology in a northern city and a southern city in China.

METHODSUsing polymerase chain reaction (PCR) and direct sequencing of HBV DNA PCR products, the serotypes and genotypes of HBV in 530 from HBsAg positive samples. The enrolled patients were from Harbin, a northern city and Lianjiang, a southern city in China.

RESULTSComparison of the serotypes and genotypes of HBV between Harbin and Lianjiang showed that adrq+ was the most predominant hepatitis B virus serotype in both Harbin and Lianjiang (87.2% and 73.5%,respectively), adw2 was the next (12.0% and 25.7%, respectively); genotype C was the most frequent in Harbin and Lianjiang (87.8% and 73.2%, respectively), and genotype B was the next (12.2% and 26.1%, respectively) only 1 patient was infected by genotype D, and 1 patient was found to be co-infected by genotype B and C in Lianjiang.

CONCLUSIONThe results suggest that the percentage of HBV serotypes and genotypes between Harbin and Lianjiang was significantly different (P less than 0.001), but the main HBV serotype and genotype of the two cities were similar.

China ; DNA, Viral ; genetics ; Genotype ; Hepatitis B Surface Antigens ; blood ; Hepatitis B virus ; classification ; genetics ; Humans ; Polymerase Chain Reaction ; Serotyping