Objective To investigate the levels of serum vascular endothelial growth factor(VEGF) and basic fibroblast growth factor(bFGF) in patients with non-small cell lung cancer(NSCLC) and relationships with c1inicopatho1ogica1 characteristics and their clinical significance. Methods The concentrations of serum VEGF and bFGF were detected by enzyme-linked immunosorbent assay(ELISA) in 40 patients with NSCLC before and after chemotherapy. Results The level of serum VEGF in patients with Ⅳ stage NSCLC was significantly higher than that of Ⅲ stage(P

Objective To evaluate effects of chronic arsenic exposure and arsenic exposure time on oxidative DNA lesions in humans. Methods A cross-sectional study was conducted in 108 subjects exposed to high concentrations of arsenic in drinking water and 75 control subjects. A cohort study was conducted in 64 subjects exposed to high levels of arsenic in drinking water for 7 or 9 years. Urinary 8-oxo-7,8-dihydredeoxygnanine(8-OHdG) levels were analyzed by the enzyme-linked immunosorbent assay kit(ELISA). Urinary arsenic concentration was detected with hydride generation atomic absorption spectroscopy. Results In the cross-sectional study, the median of urinary arsenic concentration was 484.17 mg/kg Cr for the arsenic-exposed group, and 13.80 mg/kg Cr for the control group, and the difference between the two groups was statistically significant (t=32.57, P<0.01). The median of urinary 8-OHdG levels was 16.60 and 21.88 mg/kg Cr for arsenic-exposed children and adults respectively, much higher than control children(10.50 mg/kg Cr) and adults (9.11 mg/kg Cr), and the difference was statistically significant (t=5.049, 6913, all P<0.01). Urinary 8-OHdG levels were signifieandy lower for children than adults in the exposed group(t=-1.997, P<0.05). In the cohort study, the median of urinary arsenic concentration was 461.3 mg/kg Cr for the 7-year-exposed subjects and 422.90 mg/kg Cr for the 9-year-expesed subjects, and no significant difference was observed(t=-0.250, P 0.05). The median of urinary 8- OHdG levels for 9-year-exposed children and adults were 23.46 and 24.30 mg/kg Cr respectively, significantly increased compared with those of 7-year-exposed(14.29 and 18.38 mg/kg Cr), and the difference had statical signhqcanees (t= -2.949,-3.055, all P<0.01). Conclusions Chronic arsenic exposure can lead to oxidative DNA lesions in humans. The arsenic-induced DNA lesions may aggravate with the exposure time in a certain period.

Objective To understand psychological experience of eyeball enucleation patients and provide psychological support for and analysis of patients eyeball enucleation psychological experience for clinical nursing staff.Methods Using the form of an interview,interview and analysis data of 24 eyeball enucleation patients.Results Psychological experience of patients with eyeball enucleation mainly include:(1)the desperation and fear;(2)self-image disorder;(3)care and support needs;(4)do righteousness eye demand.Conclusions Patients with eyeball take off have special psychological experience.Nursing staff should pay attention to its inner experiences adopts nursing measures to improve disease cognitive level,reduce the psychological burden and improve its postoperative quality of life.

Glycosylation is vital for activity, higher structure and function of protein. Glycoproteins derived from yeast contain N-glycan of high mannose type and are usually hyperglycosylated, while those from mammalian cells contain N-glycan of hybrid or complex type. We introduced the alpha-1,2-mannosidase I (MDSI) into yeast cells, which catalyzed an essential proceeding of N-glycan structures from Man8GlcNAc2 to Man5GlcNAc2. The plasmids contained MDSI genes from Homo sapiens [HMDSI(delta185)] or Arabidopsis thaliana [ATMDSI(delta48)], and three ER-signals were used to be transformed a mutant Pichia pastoris GJK01, respectively. The reporter protein HSA/GM-CSF (human serum albumin and granulocyte-macrophage colony stimulating factor fusion protein) was expressed and its N-glycans were analyzed by DSA-FACE (DNA sequencer assisted fluorophore-assisted carbohydrate electrophoresis). The plasmid contained ER-ScMnsI-ATMDSI(delta48) was expressed in Pichia pastoris, the Man5GlcNAc2 N-glycan on secreted glycoprotein HSA/GM-CSF was observed. The research reported here provided basic substrate to obtain the hybrid- and complex-type glycans in mammalian cell.
Gene Transfer Techniques ; Genetic Vectors ; genetics ; Glycoproteins ; biosynthesis ; Glycosylation ; Humans ; Mannose ; biosynthesis ; Oligosaccharides ; biosynthesis ; genetics ; Pichia ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics ; alpha-Mannosidase ; genetics

OBJECTIVETo examine the genetic polymorphisms in the promoter region of cyclooxygenase-2 ( COX-2) and evaluate their association with the risk of gastric cancer.

METHODSSingle-strand conformational polymorphism and DNA sequencing were used to screen the genetic variants of the COX-2 promoter region. Total 323 patients with gastric cancer and 646 frequency-matched controls were genotyped by polymerase chain reaction-restriction fragment length polymorphism method. Odds ratios (OR) and 95% confidence intervals (CI) were estimated by logistic regression. Haplotype frequency was estimated using Haplo. stat software.

RESULTSThree single nucleotide polymorphisms, including - 1290A > G, -1195G > A, and -765G > C were identified in the promoter of COX- 2. Case-control analysis showed an increased risk of developing gastric cancer for the -1290AG (OR 1.64; 95% CI 1.03-2.61), -1195AA (OR 2.68; 95% CI 1.65-4.33), and -765CG (OR 2.66; 95% CI 1.53-4.64) genotype carriers, respectively, compared with noncarriers. A greater risk of developing gastric cancer was observed for the A(-1290) -A(-1195) -C(-765) haplotype compared with the A(-1290) -G(-1195) -G(-765) haplotypes (OR 11.80; 95% CI 2.43-57.25).

CONCLUSIONGenetic polymorphisms in COX-2 promoter region may play a role in gastric carcinogenesis.

Adult ; Aged ; Cyclooxygenase 2 ; genetics ; Female ; Genetic Predisposition to Disease ; Haplotypes ; Humans ; Male ; Middle Aged ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Polymorphism, Single Nucleotide ; Promoter Regions, Genetic ; genetics ; Risk ; Stomach Neoplasms ; enzymology ; genetics

AIMTo analyze the chemical components in Danggui (the roots of Angelica sinensis (Oliv.) Diel).

METHODSHPLC-MS/MS was used to identify the main components in Danggui. Furthermore, the MS fragmentation regularity of the phthalides was proposed. The mobile phase of HPLC consisted of 0.5% acetic acid in water and 0.5% acetic acid in acetonitrile, analytical column was Hypersil ODS2 (250 mm x 4.6 mm, 5 microm), flow rate 1.0 mL x min(-1), injected volume 2 microL. The ionization source was ESI in positive ion mode.

RESULTSFerulic acid, nine known phthalides and one unknown phthalide derivative were tentatively identified in chromatograms based on their MS data and the comparison of their UV spectra with those published in the literatures.

CONCLUSIONThe structural information of phthalides was obtained via HPLC-MS/MS, which provides an accurate and fast method to identify the phthalides and provides more scientific information for quality control of Danggui.

4-Butyrolactone ; analogs & derivatives ; analysis ; chemistry ; Angelica sinensis ; chemistry ; Benzofurans ; analysis ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Coumaric Acids ; analysis ; chemistry ; Molecular Structure ; Phthalic Anhydrides ; analysis ; chemistry ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Reproducibility of Results ; Spectrometry, Mass, Electrospray Ionization ; methods

OBJECTIVETo construct TIMP-1 siRNA eukaryotic expression vectors and evaluate their effect on TIMP-1 mRNA expression in hepatic stellate cells.

METHODSThe combinant lone DNA with cutting sites of BamH I and Xho I enzyme according to the sequences of 447-465, 552-540 TIMP-1 of rats and nonspecific sequence were selected and cloned to pGEM-T vector and sub-cloned to pRNAT-U6.2. They were then identified by double enzyme digestion analysis and DNA sequencing. Three plasmids were transfected into T6 separately through an oligofectamine package. TIMP-1 mRNA expression was evaluated by RT-PCR.

RESULTSTargeting sequences of TIMP-1 siRNA eukaryotic expression vectors were correct. TIMP-1 mRNA expression was significantly reduced by transfecting them into the T6.

CONCLUSIONWe successfully constructed two TIMP-1 siRNA eukaryotic expression vectors and the transfected cells can significantly suppress the TIMP-1 expression.

Animals ; Cell Line ; Gene Silencing ; Genetic Vectors ; Hepatic Stellate Cells ; Plasmids ; RNA, Small Interfering ; Rats ; Tissue Inhibitor of Metalloproteinase-1 ; genetics ; Transfection

OBJECTIVETo investigate the association between methylentetrahydrofolate reductase (MTHFR) polymorphisms and risk of lung cancer.

METHODSTotally 505 cases with lung cancer and 500 frequency-matched controls were genotyped for the MTHFR C677T and A1298C polymorphisms by polymerase chain reaction-restriction fragment length polymorphism methods. The odds ratios (OR) and 95% confidence intervals (CI) were calculated using unconditional logistic regression model. Haplotype frequency was estimated using EH software.

RESULTSThe frequency of the MTHFR C677T allele in cases was significantly higher than that in controls (53.5% vs 44.9%, P < 0.001). Compared with the 677CC genotype, the 677CT and 677TT genotypes were associated with increased risk of lung cancer, with the OR being 1.43 (95% CI, 1.04-1.95) and 2.40 (95% CI, 1.61-3.59), respectively. In addition, a significant difference in the distribution of haplotype frequencies between cases and controls was observed.

CONCLUSIONFunctional polymorphism in MTHFR is associated with increased risk of lung cancer in Chinese population.

Adult ; Aged ; Case-Control Studies ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Lung Neoplasms ; enzymology ; genetics ; Male ; Methylenetetrahydrofolate Reductase (NADPH2) ; genetics ; Middle Aged ; Polymorphism, Genetic ; Risk

OBJECTIVETo investigate the association between genetic variations in methylenetetrahydrofolate reductase (MTHFR) and the risk of colorectal cancer in a Chinese population.

METHODSOne hundred and ninety-eight cases of colorectal cancer and 420 controls were genotyped for detecting MTHFR C677T and A1298C polymorphisms by PCR-RFLP methods. The adjusted odds ratios (OR) and 95% confidence intervals (CI) were calculated by using unconditional logistic regression model.

RESULTSThe frequency of the MTHFR C677T allele found among cases was significantly more frequent than that among controls (51.3% to 44.4%, P = 0.02). As compared with the 677CC genotype, the 677TT genotypes had a 67% (95% CI, 1.05 - 2.66) increased risk of colorectal cancers. However, there appeared no any correlations between the genetic variations of MTHFR and the cancer metastasis.

CONCLUSIONThese findings demonstrated that the genetic variation of MTHFR C677T should be a genetic susceptibility factor for colorectal cancer in a Chinese population.

Asian Continental Ancestry Group ; genetics ; China ; Colorectal Neoplasms ; ethnology ; genetics ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Logistic Models ; Methylenetetrahydrofolate Reductase (NADPH2) ; genetics ; Odds Ratio ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Polymorphism, Single Nucleotide ; Risk Factors

The aim of study was to investigate the killing effect of double suicide gene system mediated by retroviral vector on K562 cells in vivo and ex vivo. CDglyTK gene was transfected into PA317 cells by using lipofectamine. K562 cells were infected with viral supernatant. K562/CDglyTK cells were treated with 5-fluorocytosine (5-FC) and/or ganciclovir (GCV). Mice were randomly divided into three groups: tumor formation, tumor inhibition and tumor therapy. Each mouse was implanted with K562/CDglyTK cells or K562 cells. The results indicated that the killing effect of 5-FC in combination with GCV on K562/CDglyTK was more significant than using 5-FC or GCV alone. In vivo study showed that after being injected subcutaneously with K562 cells and K562/CDglyTK cells, there was not obvious difference in tumor formation rate of mice, 5-FC + GCV could suppress tumor formation of the K562/CDglyTK cells. After being treated with 5-FC and GCV, the median tumor volume of mice implanted with K562/CDglyTK cells decreased obviously, compared with the control group. Their median survival was significantly prolonged. It is concluded that double suicide genes are more effective for killing effect on K562 cells in vivo and in ex vivo. It may be applicable to clinical gene therapy.
Cytosine Deaminase ; genetics ; Flucytosine ; pharmacology ; Ganciclovir ; pharmacology ; Genes, Transgenic, Suicide ; genetics ; Genetic Therapy ; Genetic Vectors ; genetics ; Humans ; K562 Cells ; Protein-Tyrosine Kinases ; genetics ; Receptor Protein-Tyrosine Kinases ; biosynthesis ; genetics ; Recombinant Fusion Proteins ; genetics ; Recombination, Genetic ; Retroviridae ; genetics