This study examined the effect of P85 (a pluronic block copolymer) and microbubble (MB) ultrasound contrast agents under ultrasound irradiation on gene transfection and expression. The pEGFP plasmids that can encode enhanced green fluorescent protein (pEGFP) served as a report gene and were mixed with different concentrations of MB/0.05% (w/v) P85. Then the plasmids were transfected into human hepatoma G2 (HepG2) cells. The HepG2 cells treated with MB/P85 or without treatment were exposed to ultrasound (US parameters: 1 MHz, 1.0 W/cm(2), 20 s, 20% duty cycle). Twenty-four hours later, the transfection efficiency was assessed by fluorescence microscopy and fluorescence activated cell sorting (FACS) analysis. The cell viability was evaluated by Trypan blue exclusion test. The results showed that the gene transfection efficiency in HepG2 cells under ultrasound irradiation was significantly higher than that without ultrasound irradiation. HepG2 cells in the MB or P85 group in the absence of ultrasound expressed less amount of green fluorescent protein. The expression efficiency reached (22.14±3.06)% and the survival rate was as high as (55.73±3.32)% in the 30% MB plus P85 group. It was concluded that MB and P85 in the presence of ultrasound can enhance gene transfection and expression.

OBJECTIVE:To optimize the processing technology of rehmanniae radix praeparata. METHODS:Using transfer rates of catalpol,rehmaionoside D,acteoside,isoacteoside,polysaccharide as indexes for comprehensive score,heating tempera-ture(pressure),heating time and heating times as investigating factors,L9(34)orthogonal test was used to optimize the processing technology of rehmanniae radix praeparata,and verification test was conducted. RESULTS:The optimal processing technology of rehmanniae radix praeparata was as follow as heating temperature of 125 ℃,pressure of 150 kPa for twice,2 h every time. The comprehensive scores of 3 batches of samples were 0.6985,0.6755,0.7016 in the verification test,respectively,RSDs were less than 5%(n=3). CONCLUSIONS:Optimized processing technology is simple,stable,feasible,and can provide reference for in-dustrial production of rehmanniae radix praeparata.

OBJECTIVETo assess the association of neural precursor cell expressed developmentally down-regulated 4 (NEDD4) with schizophrenia.

METHODSFive single nucleotide polymorphisms (SNPs) of the NEDD4 gene were genotyped by TaqMan SNP genotyping assay in an independent sample of 464 individuals with schizophrenia and 487 healthy controls from eastern Han Chinese population. Clinical data were collected with a general information questionnaire and Positive and Negative Syndrome Scale (PANSS).

RESULTSFrequencies of rs3088077 (allelic: χ2=18.024, P=0.000; genotypic: χ2=16.634, P=0.000), rs7162435 (allelic: χ2=6.771, P=0.009; genotypic: χ2=7.352, P=0.025) and rs2303579 (allelic: χ2=11.253, P=0.001; genotypic: χ2=12.248, P=0.002) were found to be significant different between the two groups. Moreover, TT of rs7162435 was significantly correlated with scores of factors of excitement and hostility (14.53±3.925, F=3.551, P=0.029).

CONCLUSIONrs3088077, rs7162435 and rs2303579 of the NEDD4 gene may be associated with schizophrenia. Moreover, the TT genotype of rs7162435 may increase the severity of excitement and hostility. Our results may provide a clue for delineating the connection between the glutamate hypothesis of schizophrenia and ubiquitination.

Adolescent ; Adult ; Aged ; Alleles ; Asian Continental Ancestry Group ; ethnology ; genetics ; China ; ethnology ; Endosomal Sorting Complexes Required for Transport ; genetics ; Female ; Genetic Predisposition to Disease ; Genotype ; Humans ; Male ; Middle Aged ; Nedd4 Ubiquitin Protein Ligases ; Polymorphism, Single Nucleotide ; Schizophrenia ; enzymology ; ethnology ; genetics ; Ubiquitin-Protein Ligases ; genetics ; Young Adult

This study examined the effect of P85 (a pluronic block copolymer) and microbubble (MB)ultrasound contrast agents under ultrasound irradiation on gene transfection and expression.The pEGFP plasmids that can encode enhanced green fluorescent protein (pEGFP) served as a report gene and were mixed with different concentrations of MB/0.05％ (w/v) P85.Then the plasmids were transfected into human hepatoma G2 (HepG2) cells.The HepG2 cells treated with MB/P85 or without treatment were exposed to ultrasound (US parameters:1 MHz,1.0 W/cm2,20 s,20％ duty cycle).Twenty-four hours later,the transfection efficiency was assessed by fluorescence microscopy and fluorescence activated cell sorting (FACS) analysis.The cell viability was evaluated by Trypan blue exclusion test.The results showed that the gene transfection efficiency in HepG2 cells under ultrasound irradiation was significantly higher than that without ultrasound irradiation.HepG2 cells in the MB or P85group in the absence of ultrasound expressed less amount of green fluorescent protein.The expression efficiency reached (22.14±3.06)％ and the survival rate was as high as (55.73±3.32)％ in the 30％ MB plus P85 group.It was concluded that MB and P85 in the presence of ultrasound can enhance gene transfection and expression.