A PCR assay used for detection and quantification of Neotyphodium lolii in tissues of the grass Lolium perenne L. was developed. Compared with the Microscopic analysis of stained tissues, this method was more accurate. For quantitative applications, real time PCR was used to quantify Neotyphodium lolii in Lolium perenne L.. The results showed that there were some variations between different individuals, but no variance existed between different tillers from the same plant. Thus it can be concluded that fungal content in the plant is not only associated with species/variety specific, but also with the genotype of the host plant as well.

Adipates ; analysis ; Air Pollutants, Occupational ; analysis ; Esters ; analysis ; Gas Chromatography-Mass Spectrometry ; methods ; Phthalic Acids ; analysis ; Workplace

AlM:To investigate the effect of teaching mode of problem- based learning ( PBL ) in the teaching of medical students' clinical ophthalmology.
METHODS: Five classes ( total 148 students ) were randomly selected as experimental group, using PBL method, at the same time another 5 classes ( total 151 students) were also randomly selected as control group, using lecture-based learning ( LBL) mode in 2010 grade. The scores of the experimental group were compared with control at the end of term. ln addition, students and teachers were respectively interviewed using self -administered questionnaire to obtain their evaluation for PBL practice.
RESULTS:The mean scores of PBL group (78. 35±7. 63) were significantly higher than control group (71. 68±6. 37) (P<0. 001). Most of students thought that their ability of referring, synthesizing and analyzing information was enhanced by PBL, and their skills both in written and oral were also improved. PBL made it easier to understand the contexts of course. lt was the best way to improve the effect of teaching in ophthalmology based on the increase of quality in novitiate that gives more chance to students of contacting with practice, developing the ability of clinical thinking and verifying the theory in clinical novitiate. Lots of teachers considered that the classroom atmosphere was more active, students were becoming more and more proactive on their classes and the relationship between students and teachers were more harmonious when PBL was used.
CONCLUSlON:Using PBL teaching mode can highly improve the teaching effectiveness of clinical epidemiology, which is worth popularizing.

Breast Neoplasms ; chemistry ; genetics ; Chromogenic Compounds ; Female ; Genes, erbB-2 ; Humans ; In Situ Hybridization ; methods ; Receptor, ErbB-2 ; analysis

OBJECTIVETo observe the effect of synchronous perfusion of specific respiratory chain complex IV inhibitor sodium azide (NaN3) in brain on rat ventromedial prefrontal cortex (mPFC) and acetylcholine (ACh) and choline (Ch) contents in hippocampal extra-cellular fluid, and establish the AD rat model induced by mitochondrial acute injury.

METHODThe synchronous dual-probe dual-channel brain microdialysis sampling technology was applied to synchronously perfuse modified Ringer's solution containing NaN3 (50 micro mol L-1) and neostigmine (2 micro mol L-1) into mPFC and hippocampus of conscious, freely moving normal rats, and continuously collect dialysates from different encephalic areas. Dynamic contents of ACh and Ch were determined by high performance liquid chromatography-post-column immobilized enzyme reactor-electrochemical process.

RESULTACh and Ch contents in mPFC extracellular fluid of normal rats were higher than that in hippocampus. During the process of perfusion, NaN3 could significantly reduce ACh in mPFC/hippocampal extra-cellular fluid, but remarkably increase Ch, and constantly inhibit the recovery of ACh and Ch contents in mPFC/hippocampus.

CONCLUSIONThe synchronous perfusion of NaN3in rat mPFC and hippocampus can injure functions of the cholinergic nerve projection area, and cause the acute AD model with ACh and Ch metabolic disorders. This model can be used in pathogenetic and pharmacological studies.

Acetylcholine ; metabolism ; Animals ; Choline ; metabolism ; Extracellular Fluid ; drug effects ; metabolism ; Hippocampus ; cytology ; Male ; Neurotransmitter Agents ; metabolism ; Perfusion ; Prefrontal Cortex ; cytology ; Rats ; Rats, Sprague-Dawley ; Sodium Azide ; administration & dosage ; pharmacology ; Time Factors

Objective To analyze the clinical characteristics,muscle pathological features and pathogenic gene mutation in 4 cases with LMNA-related congenital muscular dystrophy (L-CMD).Methods Clinical data of the probands and the parents were collected.Skeletal muscle specimens were biopsied from the probands for pathological analysis.Genomic DNA and RNA were extracted from peripheral blood leukocytes,and PCR,reverse transcription(RT)-PCR and DNA direct sequencing were employed to analyze the LMNA gene to determine the gene mutation and confirm the pathogenicity.Results Four patients had symptoms from fetal period to several months after birth.They presented with motor retardation,muscle weakness with prominent the proximal upper limbs,distal lower limbs and neck extensor,hypotonia,contractures,with mild to moderate elevation of CK level.The muscle biopsies showed muscular dystrophic and with inflammatory changes,and the abnormal nuclear morphology was observed with transmission electron microscopy.Genetic analysis of them detected 4 dominant de novo mutations.Three of them had unreported pathogenic mutations.The same sites of the LMNA gene were wild type in their parents.Conclusions Four cases of L-CMD are genetically identified.Genetic counseling of the family can be possible.The patients should be considered LMNA gene mutation of they present themselves with muscle weakness with the proximal upper limbs,distal lower limbs and neck extensor,hypotonia,contractures,mild to moderate elevation of CK level,and if the biopsies show muscular dystrophic changes but also with inflammatory changes should be considered LMNA gene mutation.Genetic analysis is the most reliable method for diagnosing L-CMD.

Increasing attention has been focused on the antitumor activity of artemisinin derivatives in recent years, for artemisinin had been reported to have cytotoxic effects against HL-60, P388 and MCF-7 tumor cells. We report here the synthesis and evaluation for antitumor activity of a series of artemisinin-ether derivatives bearing tetrahydropyrrole, morpholine, piperidine, substituted piperidines and azoles with various linkers. Sixteen 10-O-substituted dihydroartemisinin derivatives were designed and synthesized, all of which have never been reported in literatures and whose antiproliferative effects on human breast cancer MCF-7, MCF-7/Adr and HL-60 cells were determined by MTT assay or direct cell counting. Each of these artemisinin derivatives possessed better effects than dihydroartemisinin evidently against HL-60 and MCF-7 cells growth, while less potent than doxorubicin. All target compounds exhibited significantly improved potency compared to DHA and doxorubicin on the doxorubicin-resistant MCF-7/Adr cells, so did they in their sensitive counterparts MCF-7 cells. Among them, compounds GF02, GH04 and ZH04 showed strong activity against these three cell lines growth. Further research is undergoing.
Antineoplastic Agents ; chemical synthesis ; chemistry ; Artemisinins ; chemical synthesis ; chemistry ; Breast Neoplasms ; pathology ; Cell Proliferation ; Doxorubicin ; Drug Design ; HL-60 Cells ; drug effects ; Humans ; MCF-7 Cells ; drug effects

Adenocarcinoma ; metabolism ; pathology ; Chromogranin A ; metabolism ; Colonic Neoplasms ; metabolism ; pathology ; Humans ; Immunohistochemistry ; methods ; Proliferating Cell Nuclear Antigen ; metabolism ; Staining and Labeling ; methods ; Vimentin ; metabolism

Objective To investigate the effect and possible signal pathway of brain - derived neurotrophic factor (BDNF) on apoptosis of rat embryo brain cells suffering from intrauterine hypoxic - ischemic injury. Methods The uterine arteries of the pregnant rats were clamped for 30 minutes in both experimental group and control group. BDNF(2 ?g) was injected into rats in experimental group while saline was injected into rats in control group through caudal veins. The samples were collected at 24, 48 and 72 hours re-spectively after artery clamping. Neuroapoptosis of different groups induced by ischemic damage was measured by TUNEL assay. The expression of extracellular signal - regulated kinase(ERK)and c - Jun - N - terminal kinase(JNK) were observed by immunohisloche-mistry.Results The number of apoptosis cells after hypoxic - ischemic injury increased progressively with time.The apoptosis cells number in experimental group were much lower in number than those of ischemic control group.The expression of ERK increased while the expression of JNK decreased in experimental group, comparing with that of the ischemic control group, with statistical signif-icance (P

Objective To evaluate the effect of different nebulization inhalation methods on blood oxygen degree of saturation(SaO2) in infants with asthma.Methods Sixty-two infants with asthma were randomly assigned into 3 groups: air-high-frequency flow stonized inhalation group(n=22),oxygen-high-frequency flow stonized inhalationthe group(n=20) and ultrason jet nebulization group(n=18).Three groups all were gived budesonide suspl.SaO2 was monitored during nubulization.Results There were significant differences of SaO2 levels between oxygen-high-frequency flow stonized inhalationthe group and air-high-frequency flow stonized inhalationthe group,ultrason jet ne-bulization group after 10 min and during inhalation(Pa