Chloroplast genetic engineering, offers several advantages over nuclear transformation, including high level of gene expression, increased biosafety, remedying some limitations associated with nuclear genetic transformation, such as gene silencing and the stability of transformed genes. It is now regarded as an attractive new transgenic technique and further development of biotechnology in agriculture. In this article we reviewed the characteristics, applications of chloroplast genetic engineering and its promising prospects were discussed.
Biotechnology ; methods ; Chloroplasts ; genetics ; Genetic Engineering ; methods ; Plants, Genetically Modified ; genetics ; Transformation, Genetic

Animals ; Electroencephalography ; Heart ; physiopathology ; Immersion ; physiopathology ; Rats ; Rats, Wistar ; Seawater

OBJECTIVE To investigate the effect of mono-2-ethylhexyl phthalate(MEHP) on proliferation of primary neural stem cells(NSCs)of rats and NE-4C cells of mice and on the migration of NE-4C cells and the mechanism. METHODS NE-4C or NSCs were treated with MEHP 1,10,100 and 1000 μmol · L-1 for 72 h,respectively. The cytotoxicity was estimated with the cell counting kit-8 (CCK-8). Cell proliferation was analyzed by EdU assay. The mRNA expression levels of the glucocorticoid receptor(GR),signal transducer and activator of transcription 3(Stat3)and sex determining region Y (SRY)-box 2(Sox2) were detected by qRT-PCR. The protein expression levels of total GR,GRβ, Sox2,Stat3 and p-Stat3 were measured by Western blotting. RESULTS Cell viability of NE-4C cells and NSCs at MEHP 1000μmol·L-1 was significantly decreased,which was 70.3%and 40.0%of the control group, respectively. EdU assay showed that MEHP 100 μmol · L-1 decreased NE-4C cells and NSCs by 74.8%and 12.0%(P<0.05)compared with control. The effect of MEHP on the cell migration of NE-4C was evidenced by the fact that the migration was obviously reduced to (63.4±2.0)%(P<0.05)after treatment with MEHP 100μmol · L-1 for 72 h. The mRNA expression levels associated with proliferation and migration in NE-4C of GR,Stat3 and Sox2 in MEHP 100 μmol · L-1 group were down-regulated to 49.8%,26.0% and 14.0%of control(P<0.05). At MEHP 100μmol · L-1,mRNA of GR, Stat3 and Sox2 in NSCs declined to 10.0%,14.0% and 15.3% of normal control. Western blotting results revealed that protein expressions of GR,GRβ,Sox2 and p-Stat3 were remarkably inhibited by MEHP 100 μmol · L-1 in that the relative expression of NE-4C was 0.92 ± 0.17,0.87 ± 0.35,0.81 ± 0.22 and 0.62 ± 0.24(P<0.05). The corresponding protein expression in NSCs was 0.82 ± 0.20,0.56 ± 0.12,0.84 ± 0.36 and 0.53 ± 0.20(P<0.05)when the cells were treated with MEHP 100μmol · L-1 for 72 h. CONCLUSION MEHP can inhibit the proliferation and migration of NE-4C cells and NSCs possibly by decreasing Stat3 and Sox2 that are mediated by GRβ.

0.05). Compared with model group, levels of DA and DOPAC in striatum reduced in Xieqing group (P

OBJECTIVETo assess the value of contrast-enhanced ultrasound imaging in monitoring the therapeutic effect of argon-helium cryosurgical treatment of malignant tumors.

METHODSBefore and after argon-helium cryosurgical treatment, 42 patients underwent contrast-enhanced ultrasound imaging, conventional ultrasound imaging and enhanced CT or magnetic resonance imaging (MRI) for examining the number of tumor foci and the size of necrotic areas.

RESULTSA total of 80 tumor lesions were detected by contrast-enhanced ultrasound imaging. Compared with conventional ultrasound imaging, contrast-enhanced ultrasound imaging detected a significantly greater number of tumors and the intratumoral necrotic areas (96 vs 19) as well as a significantly increased mean size of necrotic areas (5.7∓3.6 cm vs 2.8∓1.7 cm), showing no significant differences from the results by enhanced CT and MRI (94 and 5.5∓3.3 cm, P=0.872 and 0.978, respectively). The short-term therapeutic effect of argon-helium cryosurgery evaluated by contrast-enhanced ultrasound imaging were also similar to that assessed by enhanced CT or MRI (P=0.906).

CONCLUSIONContrast-enhanced ultrasound imaging has important values in monitoring malignant tumors during argon-helium cryosurgical treatment and in evaluating the short-term therapeutic effect of the treatment.

Adult ; Aged ; Argon ; Cryosurgery ; methods ; Female ; Helium ; Humans ; Male ; Middle Aged ; Neoplasms ; diagnostic imaging ; surgery ; Treatment Outcome ; Ultrasonography

OBJECTIVETo detect and compare the transcriptional activities of prostate-specific membrane antigen (PSMA) promoter and enhancer and survivin promoter in different human prostate cancer cell lines, and to search for some evidence for the targeting gene therapy of human prostate cancer.

METHODSThe fragments of the PSMA promoter and enhancer and survivin promoter were amplified by PCR and inserted into pGL3-Basic. The recombinant plasmids were transiently transfected into human prostate cancer cell lines and normal Chang liver cells, and, their transcriptional activities in various cells were determined by measuring the expression of luciferase.

RESULTSThe survivin promoter exhibited a higher transcriptional activity than PSMA promoter and enhancer in tumor cell lines, and the S2pro promoter showed the highest activity, reaching one third of that of the CMV promoter.

CONCLUSIONThe survivin promoter is highly activated in prostate cancer cell lines and may serve as a new tool for the transcriptional targeting gene therapy of prostate cancer.

Antigens, Surface ; genetics ; Cell Line, Tumor ; Glutamate Carboxypeptidase II ; genetics ; Humans ; Inhibitor of Apoptosis Proteins ; genetics ; Male ; Plasmids ; Promoter Regions, Genetic ; Prostatic Neoplasms ; genetics ; therapy ; Transcription Initiation Site ; Transcriptional Activation ; Transfection

The present study is to determine the flavonoid glycosides, terpene lactones, biflavones, gingko acid and procyanidins of ginkgo pollen. UPLC-TQ-MS technology was used for the determination of 24 kinds of resource chemical composition in ginkgo pollen qualitatively and quantitatively. The results shows that the contents of rutin, quercetion 3-O-[4-O-(α-L-rhamnosyl )-β-D-glucoside] and kaempferolis were 120.9, 114.0, 222.1 μg x g(-1). In this paper, the contents of 24 kinds of chemical components of ginkgo pollen were determinated by UPLC-TQ-MS for the first time. This method is simple and quick, which will be benefit for recycling utilization of ginkgo pollen.
Chromatography, High Pressure Liquid ; methods ; Flavonoids ; analysis ; Ginkgo biloba ; chemistry ; Mass Spectrometry ; Pollen ; chemistry ; Proanthocyanidins ; analysis ; Rutin ; analysis ; Terpenes ; analysis

OBJECTIVETo clone DNA sequence of the survivin promoter and study is transcriptional activities in human prostate cancer cells and normal Chang liver cells.

METHODSThe fragment of the survivin promoter was acquired by PCR amplification and inserted into pPRIME vectors to reconstruct a recombinant plasmid named pPRIME-S1pro and pPRIME-S2pro. Then the reconstructed plasmid was transiently transfected into human prostate cancer cells lines LNCaP and normal Chang liver cells. The transcriptional activities of the survivin promoter in various cells was determined by measuring the expression of green fluorescent protein (GFP).

RESULTSThe survivin promoter had transcriptional activities in LNCaP cells and the transcriptional activity of the S2pro was much higher that of the S1pro, reaching a level of 39% of the transcriptional activity of the CMV promoter.

CONCLUSIONThe survivin promoter cloned in the therapy for prostate cancer.

Cell Line, Tumor ; Humans ; Inhibitor of Apoptosis Proteins ; Male ; Microtubule-Associated Proteins ; biosynthesis ; genetics ; Neoplasm Proteins ; biosynthesis ; genetics ; Plasmids ; Polymerase Chain Reaction ; Promoter Regions, Genetic ; Prostatic Neoplasms ; genetics ; metabolism ; Transfection

Objective MR microscopy technique was used to study the visualization of senile plaque deposition in brains of the Alzheimer disease(AD)transgenic mice.Methods Two transgenic mice and 2 wild type mice at the age of 17 months were scanned in vivo using T_2 weighted image.After MR imaging,the brains were cut serially and immunostained according to the orthogonal pilot images.MR T_2 weighted images and immunohistological images of the senile plaque were observed and matched.Results The MR images showed that some black spots were visible in the hippocampus and cerebral cortex of the AD transgenic mice and some spots were consistent with the senile plaques on immunohistological sections.There were no spots in the MR images and the immunohistological sections of the wild type mice.Conclusion It is possible that MR microscopy can be used to detect the deposition of the senile plaque and diagnose AD specifically.

OBJECTIVETo investigate the correlation between the polymorphism of dopamine beta hydroxylase(DBH) gene and the susceptibility of Shanghai Chinese Han population to Parkinson's disease(PD).

METHODSAssociation study was performed in 144 PD patients and 188 healthy control subjects matched for age, sex and origin. Polymorphism of DBH gene was analyzed with polymerase chain reaction-restriction fragment length polymorphism.

RESULTSThe allelic frequency of A2 allele of DBH gene was significantly higher in PD patients than in controls(P<0.01).The risk of suffering from PD increased (OR=1.82) in the individual with A2 allele. And the genotypic frequency of A2/A2 was significantly higher in PD patients(OR=2.11, P<0.01),too. On the other hand, the allelic frequency of A1 allele and the genotypic frequency of A1/A2 genotype of DBH gene in PD patients were significantly lower(A1 alleles: OR=0.54, P<0.01; A1/A2 genotypes: OR=0.45, P<0.01).

CONCLUSIONThe polymorphism in DBH gene might play an important role in the susceptibility of Shanghai Chinese Han population to PD.

Adult ; Aged ; Asian Continental Ancestry Group ; genetics ; China ; Dopamine beta-Hydroxylase ; genetics ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; Humans ; Male ; Middle Aged ; Parkinson Disease ; genetics ; Polymorphism, Genetic ; Polymorphism, Restriction Fragment Length