Effect of intermittent high glucose on oxygen-glucose deprivation/refurnish neuronal survival / 中华危重病急救医学

Objective To investigate the effect of intermittent high glucose on oxygen-glucose deprivation/refurnish (OGD/R) neuronal survival.Methods The primary cultured hippocampal neurons of mice were sub-cultured when the cell fusion reached about 80％.Cells in logarithmic growth phase were placed in a hypoxic incubator (37 ℃,5％ CO2,95％ N2) to simulate cell hypoxia.The culture medium was replaced by glucose-free Hank equilibrium salt solution (HBSS) to simulate cell hypoglycemia.The normal glucose and oxygen control group was set up.Cell morphology was observed under inverted phase contrast microscope after 6 hours of hypoxia and hypoglycemia treatment,and cell viability was detected by CCK-8 cell proliferation assay kit,and then grouping experiment was carried out.The cells were randomly divided into four groups.The cells were cultured in different concentration glucose medium under normal oxygen,5％ CO2 and 37 ℃ for 72 hours to prepare OGD/R model of cell ischemia/reperfusion.The low-glucose control group was cultured in medium containing 5.5 mmoFL glucose.The constant high-glucose group was cultured in medium containing 33.0 mmol/L glucose.The intermittent high-glucose group was cultured in medium containing 33.0 mmol/L glucose for 3 hours then in medium containing 5.5 mmol/L glucose for 2 hours alternately for 3 times during the day,and overnight in medium containing 33.0 mmol/L glucose at night.The hyperosmotic control group was made up of 5.5 mmol/L glucose medium and mannitol.The osmotic pressure was the same as that of the constant high-glucose group,and the effective glucose concentration was the same as that of the normal glucose and oxygen group,so as to eliminate the effect of osmotic pressure changes caused by the high-glucose medium on the results.Cell morphology was observed under inverted phase contrast microscope after 72 hours of cell culture in each group.Cell viability was measured by CCK-8 kit,and apoptotic rate was measured by flow cytometry.Results The inverted phase contrast microscope showed that the cells in the normal glucose and oxygen control group were plump and refractive,and had obvious nucleus,clear processes and high cell activity.After 6 hours of hypoxia and hypoglycemia treatment,the cells were shrunk,refractive index was poor,the nucleus was unclear,the processes were not clear,and the cell activity was significantly lower than that of normal glucose and oxygen control group (A value:0.34±0.06 vs.1.09±0.06,P ＜ 0.01),which indicated that the model of oxygen-glucose deprivation (OGD) was successfully prepared.After 72 hours of culture with different concentrations of glucose,the cells in the low-glucose control group were shrunk,the cell membrane was incomplete,the nucleus was unclear,and number of necrotic cells were more.In the constant high-glucose group,the refractive index of cells was poor,a large number of cells floated,and the nucleus was not obvious.In the intermittent high-glucose group,the cell morphology was normal,the refractive rate of cells was decreased slightly,and the necrotic cells were less.In the hypertonic control group,the cell status was close to that in the constant high-glucose group.Compared with the low-glucose control group or constant high-glucose group,the cell viability in the intermittent high-glucose group was significantly increased (A value:2.04±0.15 vs.0.64±0.18,1.16±0.16,both P ＜ 0.01),the apoptotic rate was significantly decreased [(59.60 ± 2.55)％ vs.(78.15 ± 15.77)％,(95.60± 0.14)％,both P ＜ 0.05].There was no significant difference in cell activity or apoptotic rate between the hypertonic control group and the constant high-glucose group [cell activity (A value):1.07 ± 0.07 vs.1.16 ± 0.16,apoptotic rate:(87.80 ± 4.53)％ vs.(95.60 ± 0.14)％,both P ＞ 0.05].Conclusion Intermittent high glucose within a certain range had protective effect on OGD/R neuronal survival.

Refractive development during the past 5 years among primary school students in Jinshan District of Shanghai / 中国学校卫生

Objective@#To explore refractive development of primary school students in Jinshan district of Shanghai during the past five years.@*Methods@#A total of 201 first grade pupils enrolled in 2013 were followed up for 5 years. Annual examination of non-cycloplegic refraction and axial length (AL) was implemented and analyzed.@*Results@#The mean spherical equivalents (SEs) of boys at each grade was (0.22±0.53)(-0.04±0.64)(-0.36±0.92)(-0.74±1.23)(-1.14±1.67)D, respectively; for girls, (0.26±0.88)(-0.03±1.02)(-0.28±1.02)(-0.64±1.32)(-1.13±1.65)D, respectively. The mean ALs of boys at each grade was (22.94±0.60)(23.13±0.68)(23.45±0.69)(23.65±0.81)(24.03±0.93)mm, respectively and was (22.40±0.67)(22.67±0.70)(22.95±0.74)(23.14±0.79)(23.59±0.90)mm for girls at each grade, respectively. There were negative correlations between dioptres and ALs in each grade(r=-0.26, -0.35, -0.41, -0.53, -0.59, P<0.05).@*Conclusion@#The dioptre and AL among primary school students in Jinshan district of Shanghai increased gradually and results in developing into myopia. The dioptre negatively associates with AL, which should be both paid attention to among primary school students.

Study on resistance mechanisms and epidemiology of Enterobacteriaceae with decreased carbapenem susceptibility / 中华微生物学和免疫学杂志

Objective To investigate the resistance mechanisms and epidemiology of Enterobacteriaceae isolated from clinical with reduced susceptibility to imipenem or meropenem.Methods 18 strains of Enterobacteriaceae with reduced carbapenem susceptibility were collected during January to August in 2010.The MICs of these strains were determined using automated microbial identification system.ESBLs,AmpC and KPC were tested using the agar dilution method.PCR amplification and DNA sequence were performed to analyze the KPC genes,PFGE was used to examine the molecular epidemiology.Results All 18 strains were detected ESBLs and AmpC,14 strains were detected KPC-2.3 strains with EDTA paper method positive may produce other metal carbapenem,in which 2 strains harbor KPC-2.PFGE types indicate that there were six genotypes among 15 strains of Klebsiella pneumoniae.Conclusion Plasmid-mediated KPC-2 was the main reason which makes Enterobacteriaceae reducing carbapenem susceptibility and causes short-term epidemic in hospital.Clinical strains harboring KPC-2 gene may carry multiple resistance genes meanwhile.

ESBLs′ Genotypes of Escherichia coli and Klebsiella pneumoniae / 中华医院感染学杂志

OBJECTIVE To investigate the ESBLs ′ genotypes and the resistance of Escherichia coli and Klebsiella pneumoniae in Taizhou Hospital of Zhejiang Province.METHODS We collected 30 strains of E.coli and K.pneumoniae expressing ESBLs,then analyzed their encoding genotypes of TEM,SHV,PER,VEB and CTX genes by PCR and DNA sequencing technology separately.RESULTS There were 26 strains expressing blaCTS-M gene among 30 strains,in which 16 strains belonged to blaCTS-M-9 subgroup genotype,8 strains to blaCTX-M-1,7 strains to blaCTX-M-2,4 strains to blaCTX-M-1 as well as blaCTX-M-2,1 strain to blaCTX-M-2 as well as blaCTX-M-9 and 1 strain belonged to blaCTX-M-1 as well as blaCTX-M-9.CONCLUSIONS The prevalent clinical genotype of E.coli and K.pneumoniae is blaCTX-M-9.

Detection of CTP gene expression in the asexual erythrocytic stages of Plasmodium falciparum (FCC1 /HN) by reverse transcriptase- polymerase chain reaction and construction of eukaryotic expression vector of CTP gene / 热带医学杂志

Objectve To detect whether the CTP(phosphocholine cytidylyltransferase) gene was expressed in the asexual erythrocytic stages of Plasmodium falciparum (FCC 1/HN )by using the RT - PCR and to construct eukaryotic expression vector of CTP. Method The erythrocytic stage parasites of Plasmodium falciparum were cultured as described by Trager and Jensen. RNA from erythrocytic stage parasite was extracted by using Trizol reagent. The complete genes coding for CTP gene isolates FCCI/HN were amplified by reverse transcriptase -polymerase chain reaction(RT- PCR). CTP gene was cloned into eukaryotic expression vector pcDNA3. Results CTP encoding gene was amplified from the erythrocytic stages of Plasmodiumfalciparum (FCC 1/HN) and eukaryotic expression vector of CTP was constructed. Conclusion CTP gene was expressed in the erythrocytic stages of Plasmodium falciparum (FCC 1/HN) and eukaryotic expression vector of CTP was successfully constructed.

Association Rules of Chinese Herbal Compound Formula for Osteoporosis / 中国药房

OBJECTIVE:To discuss the medication regularity of Chinese herbal compound formula for osteoporosis. METHODS:A total of 104 Chinese herbal compound formula for osteoporosis (OP) published from 1995 to 2005 were retrieved from China Hospital Knowledge Database(CHKD); and a database was set up using EXCEL2000 software,with the drug name of each herbal medicine in the compound herbal formula included and a statistical analysis was carried out.The herbs which were used frequently were subjected to data mining and analysis using association rules. RESULTS:Of the total 104 Chinese herbal compound formula for osteoporosis,leading the first 12 places in terms of application frequency in turn were prepared Rehmannia glutinosa,Herba Epimedii,Dioscorea opposita and Eucommia ulmoides,etc.In the association rule set,there were respectively 7,6,6 and 8 rules which respectively consisted of 2,3,4 and(5,6,7) herbs.CONCLUSION:The relationship between the three factors(renal deficiency,splenic asthenia,and blood deficiency) and osteoporosis were verified in the study of the Chinese herbal compound formula for osteoporosis.

Construction of Prokaryotic Expression Vector of the Fusion Gene IFN-?1b/CSPⅡ and Expression in E.coli / 中国寄生虫学与寄生虫病杂志

Objective To Construct the prokaryotic expression vector of the fusion gene IFN-?1b/CSPⅡ. MethodsIFN-?1b was amplified from the human genomic DNA by PCR and cloned into prokaryotic expression vector pGEX-4T-1. The recombinant plasmid pGEX-4T-1/IFN-?1b was constructed. Circumsporozoite proteinⅡ(CSPⅡ) was amplified from the Plasmodium falciparum genomic DNA by PCR and was cloned into the prokaryotic expression vector pGEX-4T-1. The recombinant plasmid pGEX-4T-1/CSPⅡ was constructed. IFN-?1b was cut from the recombinant plasmid pGEX-4T-1/IFN-?1b digested with BamHⅠ and EcoRⅠ and ligated with the recombinant plasmid pGEX-4T-1/CSPⅡ also digested with BamHⅠ and EcoRⅠ . The recombinant prokaryotic plasmid pGEX-4T-1/IFN-?1b/CSPⅡ was constructed. The fusion gene IFN-?1b/CSPⅡ was expressed in E.coli by IPTG. Results The prokaryotic expression vector pGEX-4T-1/IFN-?1b, pGEX-4T-1/CSPⅡ and pGEX-4T-1/IFN-?1b/CSPⅡ were identified by PCR, enzyme digestion and gene sequencing. The expressed fusion protein/IFN-?1b/CSPⅡ in E.coli was identified by SDS-PAGE and Western blot. Conclusion The prokaryotic expression vector of the fusion gene IFN-?1b/CSPⅡ was successfully constructed, which was then expressed in E.coli .