OBJECTIVETo investigate the effect of extracellular regulating kinase (ERK) signaling pathway on the secretion of gamma-aminobutyric acid (GABA) in cultured rat hippocampal neurons induced by stromal cell derived factor-1 (SDF-1).

METHODSThe hippocampal neurons of newborn SD rats were cultured and identified in vitro; the phosphorylation level of ERK1/2 was examined by Western blot; ELISA was used to detect the effect of PD98059, a ERK1/2 specific blocker on GABA secretion of cultured hippocampal neurons and Western blot were adopted to measure the protein expression levels of glutamate decarboxylase (GAD65/67) and gamma aminobutyric acid transporter (GAT); after blocking ERK1/2 signaling pathway with PD98059; RT-PCR was used to detect the mRNA expression levels of GAT-1 and GAD65 after treated with PD98059.

RESULTSThe levels of ERKl/2 phosphorylation were increased significantly by SDF1 acting on hippocampal neurons, and CX-CR4 receptor blocker AMD3100, could inhibit SDF-1 induced ERK1/2 activation; SDF-1 could inhibit the secretion of GABA in cultured hippocampal neurons, and ERK1/2 specific inhibitor PD98059, could partly reverse the inhibition of GABA secretion by SDF-1. The effects of SDF-1 on cultured hippocampal neurons was to decrease the mRNA genesis of glutamic acid decarboxylase GAD65 and GABA transporter GAT-1, besides, ERK inhibitor PD98059 could effectively flip the effect of SDF-1. The results of Western blot showed that SDF-1 could inhibit the protein expression of GAT-1 and GAD65/67 in hippocampal neurons and the inhibition of GAT-1 and GAD65/67 protein expression could be partially restored by ERK1/2 blocker.

CONCLUSIONSDF-1 acts on the CXCR4 of hippocampal neurons in vitro, and inhibits the expression of GAD by activating the ERK1/2 signaling pathway, and this may represent one possible pathway of GABA secretion inhibition.

Animals ; Blotting, Western ; Cells, Cultured ; Chemokine CXCL12 ; pharmacology ; Flavonoids ; pharmacology ; Glutamate Decarboxylase ; metabolism ; Hippocampus ; cytology ; MAP Kinase Signaling System ; Neurons ; metabolism ; Phosphorylation ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Receptors, CXCR4 ; metabolism ; gamma-Aminobutyric Acid ; secretion

Objective To analyze the assessment results of iodine deficiency disorders(IDD) External Quality Control Laboratories at all levels in Guizhou province and the network operation to further standardize and improve the laboratory, improve network performance, provide reliable laboratory quality assurance for epidemiological surveillance and control of IDD and reliable decision-making. Methods The results of different level of IDD labs in Guizhou that took part in the entire country's IDD lab exo-network quality control examination of 1999 - 2008, which organized by the nation iodine deficiency disorder reference lab were analyzed. Results In the past 10 years, except the provincial laboratory examination results of urinary iodine in 2000 was failed, the other results were all qualified in the rest 9 years;iodized salt examination results were qualified. The urinary iodine laboratory response rate of Guizhou provinces and municipalities(state, district), from 1999 to 2002, were 88.9%(8/9), 66.7%(6/9), 77.8%(7/9), 66.7%(6/9), respectively, and the rate was stable at 100% from 2003 to 2008. Qualifying rate reached 100% in 2007, the remaining 9 years were 33.0%(3/9) - 88.9%(8/9). The iodized salt laboratory response rates were 100%(9/9) in 2000 - 2008. The pass rates were 77.8%(7/9), 88.9%(8/9),77.8%(7/9) from 2001 to 2003, 88.9%(8/9) in 2007, and the remaining 5 years 100%. Response rate of iodized salt laboratory at the county level that participated in the External Quality Control were 66.7% (20/30), 90.0%(27/30), 80.0% (24/30), 96.7% (29/30) from 2000 to 2003, respectively, and 2006 was 96.7% (29/30), and the remaining four years were all 100% (30/30). The pass rates in 2000 - 2008 were between 53.3% (16/30) -93.3%(28/30). Conclusions The accuracy of test results of external quality controls and the normal operation of the network at all levels of laboratories is closely related to the IDD laboratory conditions, detection techniques, and the degree of attention of relevant departments and professional bodies.

Objective To evaluate the development of immature oocytes after freezing-thawing by conventional cryopreservation method for mature oocytes.Methods Immature oocytes were collected from stimulated ovaries of intracytoplasmic sperm injection(ICSI)cycles.Immature oocytes were in vitro matured directly or after slow freezing-fast thawing and immunostained for tubulin and chromatin and at last visualized by confocal microscopy.Results No statistical difference was found in maturity rate between freezing groups and the controls.There was a statistically significant increase in abnormalities of chromosome(23.7% vs. 50%)and spindle(28.9% vs.53.9%)in the GV freezing group compared with the GV control(P

Objective To compare the effects of vitrification with slow-freezing on the developmental ability of day 3 cleavage stage embryos.Methods Patients who had no less than 4 high quality embryos were included in this study.These embryos were cryopreserved using the methods of vitrification or slow-freezing.In the eryopreserved embryo transfer cycles,the embryos which were cryopreserved using one of the methods were chosen randomly.The developmental ability of embryos was compared between these two groups.Results A total of 80 patients were included in this study with 160 embryos.In the group of slow-freezing,73(91%)embryos were survived and achieved 15(38%)clinical pregnancies.Among these,3 were twins and the implantation rate was 25%(18/73).In the group of vitrification,71(89%)embryos were survived and achieved 19(48%)clinical pregnancies.Among these, 9 were twins and the implantation rate was 39%(28/71),which was significantly higher than the slow- freezing group(P

Background The condition of the sclera is associated with many ocular diseases.The measurement of human scleral thickness in vivo is helpful for us to understand the features of the sclera and related diseases.Objective The present study was to measure the anterior sclera thickness(AST) in patients with senile cataract and to analyze the relationship among AST and other associated ocular parameters.Methods This study was approved by the Ethic Commission of First Hospital of Peking University.Written informed consent was obtained from each individual prior to examination.One hundred and five senile cataract patients were recruited in this study.Central corneal thickness (CCT),corneal curvature (CCV) and axial length were measured using ultrasonic pachymeter,keratometer,and A-scan unit,respectively.The AST was measured at 2 mm posterior to the scleral spur in the temporal meridian using ultrasound biomicroscope (UBM).The differences of CCT,CCV,ocular axial length and AST between bilateral eyes and the different sexes were compared by the Paired test and independent sample t test.The correlations among various parameters were assessed by the Pearson linear correlation analysis.The differences of CCT and AST among different axial length groups were evaluated by one-way ANOVA.Results No significant differences were found in the CCT,CCV,axial length and AST between bilateral eyes (t =0.584,P =0.561 ; t =1.161,P =0.248 ; t =0.140,P =0.889 ; t =0.342,P =0.773).Temporal AST at 2 mm posterior to the sclera spur was (0.589 ±0.051)mm in the right eyes.An insignificant decline in CCT was found in the male group compared with the female group (right eyes:t =0.469,P =0.641 ; left eyes:t =0.465,P =0.643).However,compared with the female group,the increase of axial length,reduction of the mean CCV value and enhancement of the mean AST were observed(right eyes:all P＜0.01 ;left eyes:all P＜0.01).CCV showed a negative correlation with ocular axial length (r =-0.50,P＜0.01),but no significant correlation was found among age,CCT,ocular axial length and AST(P＞0.05).No remarkable differences were found in CCT and AST among the various axial length groups (CCT:F =0.998,P=0.372;AST:F=1.919,P=0.383).Conclusions In senile cataract patients,correlation is not found between AST and CCT;the increase of axial length is not associated with the thinning of the eyeball wall to a certain extent.Differences exist in some ocular parameters between different sexes.

Background A close relation between sclera thickness and glaucoma has been determined.Clinical features vary in different types of glaucoma patients,which hints that the scleral thickness might be distinct among these patients.Objective The aim of this study was to investigate the differences of central corneal thickness(CCT),anterior scleral thickness(AST) and axial length in glaucomatous patients.Methods This study was approved by the Ethic Commission of First Hospital of Peking University.Written informed consent was obtained from each individual prior to the examination.A retrospective descriptive study was designed.One hundred and sixty consecutive patients were recruited from March,2009 to November,2010 in First Hospital of Peking University,including 35 eyes with primary angle closure glaucoma(PACG) (35 cases),34 eyes with primary open-angle glaucoma POAG) (34 cases),37 eyes with normal tension glaucoma(NTG) (37 cases)and 17 eyes of ocular hypertension OHT) (17 cases).Thirty-seven eyes of 37 subjects with incipient cataract served as the control group.CCT and ocular axial length were measured with ultrasonic pachymeter and A-scan unit,respectively,and AST at the temporal quadrant 2 mm posterior to the scleral spur was measured by ultrasonic biomicroscopy (UBM).The measuring parameters among different groups were compared by analysis of covariance,and the correlations of AST,CCT with ocular axial length were analyzed using Pearson linear correlation and linear regression.The differences and correlation of CCT,AST and AL among five groups were analyzed.Results The CCT values in PACG group,POAG group,NTG group,OHT group and control group were (535.54 ± 5.20),(550.47 ± 5.28),(521.61 ± 5.07),(575.75 ± 7.76) and (535.06± 5.06) μm,respectively,showing a significant difference among them (F =9.560,P =0.000),and the CCT value of OHT group was increased in comparison with POAG group,PACG group,NTG group and control group(all P =0.000).The CCT of the POAG group was thicker than that in the PACG group,NTG group and control group(P=0.046,0.000,0.040).No significant difference was found in CCT among NTG group,PACG group and control group(P=0.950,0.060).The AST values of PACG group,POAG group,NTG group,OHT group and control group were(0.593±0.050),(0.600±0.050),(0.592-±0.060),(0.610-±0.060) and(0.604±0.060) mm,respectively,showing a insignificant difference among them (F =0.700,P =0.590).The axial length in the patients with PACG was shorter than that of the POAG group,NTG group,OHT group and control group (all P =0.000).The Pearson correlation analysis showed significantly positive correlation between CCT and AST in POAG group and NTG group(r=0.445,P=0.008;r=0.400,P=0.014).Conclusions This study confirms that there is dissimilarity in CCT but not in AST among different types of glaucomatous patients.The changes of CCT and AST are consistent in POAG and NTG patients.

Objective To determine the changes of interleukin-12(IL-12) and IL-12 mRNA in gastric mucosa of children with helicobacer pylori (Hp) infection,and to study the effects of Hp infection on the expression levels of IL-12 and IL-12 mRNA,and to evaluate its possible roles in the pathogenesis of gastric mucosal inflammation in Hp related gastroduodenal diseases.Methods Biopsy specimens were taken from the antral mucosa on endoscopy in patients with or without Hp infection, which were diagnosed by urease test and Giemsa staining. The expression levels of IL-12 and IL-12 mRNA in gastric mucosa were determined by ELISA and RT-PCR.Results Inflammation of gastric antral mucosa was more severe in Hp-positive mucosa .The expression levels of IL-12 and IL-12 mRNA in Hp-positive mucosa were (66.42?15.15) ng/g and (59.21?15.03)%,which were more than those in (Hp-negative )(22.22?8.79) ng/g and (17.94?7.39)%(P

AIMTo observe the damages induced by hydrogen peroxide in cultured bovine cerebral microvascular endothelial cells (BCMEC) and evaluate the protective effects of hydroxyethylpuerarin on hydrogen peroxide-injured BCMEC.

METHODSBCMEC were cultured and transferred into modified Eagle medium (MEM). The viability of cells was detected by MTT assay. Cell injury was determined by lactate dehydrogenase (LDH) activity in the extracellular medium. Flow cytometry was employed to observe the occurrence of apoptosis. Morphologic changes of cells were visualized under phase contrast and electron microscopes.

RESULTSHydrogen peroxide (200 micromol x L(-1) for 4 hours) inhibited the viability of cultured BCMEC and stimulated LDH release. Hydrogen peroxide (100 micromol x L(-1) for 4 hours) induced the occurrence of apoptosis. Hydroxyethylpuerarin was shown to increase the survival rate and decrease the activity of LDH of BCMEC damaged by hydrogen peroxide. Hydroxyethylpuerarin was also found to protect BCMEC against apoptosis induced by hydrogen peroxide.

CONCLUSIONHydrogen peroxide induces BCMEC injury either by apoptosis or through necrosis. Hydroxyethylpuerarin protects BCMEC against hydrogen peroxide-induced injury in a concentration-dependent manner. Its antioxidant effects might be involved as the mechanism protection.

Animals ; Antioxidants ; administration & dosage ; isolation & purification ; pharmacology ; Apoptosis ; drug effects ; Brain ; blood supply ; Cattle ; Cell Survival ; drug effects ; Cells, Cultured ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Endothelial Cells ; drug effects ; metabolism ; Hydrogen Peroxide ; toxicity ; Isoflavones ; administration & dosage ; isolation & purification ; pharmacology ; Microcirculation ; drug effects ; metabolism ; Neuroprotective Agents ; administration & dosage ; isolation & purification ; pharmacology ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Pueraria ; chemistry

Base Sequence ; Conjugation, Genetic ; Drug Resistance, Bacterial ; Electrophoresis, Gel, Pulsed-Field ; Escherichia coli ; drug effects ; enzymology ; Humans ; Klebsiella pneumoniae ; drug effects ; enzymology ; Microbial Sensitivity Tests ; Molecular Sequence Data ; Polymerase Chain Reaction ; Transformation, Bacterial ; beta-Lactamases ; genetics

To develop an analytic method for qualitative and quantitative analysis of dodecatetraenamides A, B in 42 samples of two official species of Asari Radix et Rhizoma( ARR) (37 samples of Asarum heterotropoides var. mandshuricum with different collection time and 5 samples of Asarum sieboldiivar. seoulense). The HPLC-IT-TOF-MS/MS methods for the qualitative and UPLC-PDA methods for the quantitative analysis were established. Dodecatetraenamides A, B were identified by comparing the retention time, UV absorption spectrum and quasi-molecular ion peak [ M + H]+ with the reference compound using HPLC-IT-TOF-MS/MS. The content of dodecatetraenamides A and B in ARR were determined by UPLC-PDA. The separation was successfully carried out on a ACQUITY UPLC BEH C18 (2.1 mm x 100 mm, 1.7 µm) column eluted with mobile phases of water (A) and acetonitrile (B) in gradient program (0-3 min, 35% B; 3-5 min, 35%-36% B; 5-6 min, 36%-43% B; 6 min-11 min 43% B; 11-12 min, 43%-100% B). The column temperature was 45 °C, and the detection wavelength was set at 254 nm. The flow rate was 0.6 mL · min(-1). On one level mass spectrometry scanning, the results showed that the quasi-molecular ion [M + H] + of both dodecatetraenamides A and B were m/z 248.20. The quantitative method with UPLC-PDA has made the baseline separation of the constituents, which were reported as mixtures in the most literatures. The average recovery of dodecatetraenamides A and B were 97.90% and 99.86%, the relative standard deviation were 0.4% and 1.1%, respectively. The contents of dodecatetraenamides A, B in all ARR samples was in the range of 0.11-3.89 and 0.24-6.65 mg · g(-1). Their contents reduced with the extension of storage time. Compared with the samples of 2013, the average content of the two constituents in the samples collected in year 2002-2003 reduced 34% and 36%, respectively (P < 0.05). Compared the A. sieboldii var. seoulense and A. heterotropoides var. mandshuricum with the same collective time and production area, the average contents of the two constituents in latter were up to (1.59 ± 0.75) mg · g(-1) and (2.90 ± 1.17) mg · g(-1), respectively, significantly higher than that in A. sieboldii var. seoulense (dodecatetraenamide A were (0.78 ± 0.52) mg · g(-1), dodecatetraenamide B were (1.69 ± 0.83) mg · g(-1)) (P < 0.05). The content of the dodecatetraenamide A in overground part was in the range of 0.11-0.33 mg · g(-1), dodecatetraenamide B was 0. 24-0.60 mg · g(-1), which were much lower than that of the underground part of ARR (dodecatetraenamide A was in the range of 0.73-3.89 mg · g(-1), dodecatetraenamide B was 2.11-6.24 mg · g(-1)). The method was certified to be simple, accurate and reliable and could be used for qualitative and quantitative analysis of dodecatetraenamide A and B in different species of ARR, also can be used for the comprehensive quality control of traditional Chinese medicine, Asari Radix et Rhizoma.
Amides ; chemistry ; Asarum ; chemistry ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Mass Spectrometry ; Molecular Structure ; Rhizome ; chemistry