Objective To investigate the single nucleotide polymorphism(SNP)in the coding region in cyto-chrome P4501 A1 (CYP1A1)gene and to evaluate the contributions of SNPs to acute lymphocytic leukemia(ALL)and acute myeloid leukemia(AML)susceptibility in children.Methods One hundred and twenty -one(male 76,female 45)children with acute leukemia were selected from Department of Hematology in Shenzhen Children′s Hospital,Zuyi Medical College between January 2007 and January 201 4 as the case study group,and the average age was 4.42 years old.Case study group included 1 01 (male 65,female 36,average age 4.38 years old)ALL children (ALL group)and 20(male 1 1 ,female 9,average age 4.66 years old)AML children(AML group).A total of 1 1 6(male 74,female 42) children with respiratory tract infections on health examination during the same period were selected as the control group and the average age was 3.93 years old.SNPs in the coding region in CYP1A1 gene were detected by reverse transcrip-tional(RT)-PCR -denaturing gradient gel elelctrphoresis(DGGE)combined with direct sequencing in the case study group and the control group.Results Only one SNP,A4889G,was screened in the open reading frame (ORF)of CYP1A1 gene in Chinese Han children and the G allele frequency of CYP1A1 gene in the case group,ALL group,AML group and the control group were 31 .4%,32.2%,27.5% and 38.8%.The CYP1A1 A4889G AG and AG +GG geno-type were linked with decreased risk of AML(OR =0.31 ,95%CI:0.1 1 -0.87,P =0.02;OR =0.35,95%CI:0.1 4 -0.93,P =0.03)especially in boys with AML(OR =0.1 2,95%CI:0.03 -0.60,P =0.01 ;OR =0.1 6,95%CI:0.04 -0.65,P =0.01 ),but the CYP1A1 A4889G polymorphism was not associated with the risk of ALL(P >0.05). The CYP1A1 A4889G allele frequency and the distribution of genotypes were significantly different from those reported in America,India,Korea,Brazil and Iran(all P <0.05).Conclusions CYP1A1 A4889G polymorphism may be not as-sociated with susceptibility to ALL,but may decrease the risk of childhood AML especially in boys with AML.In addi-tion,it may exhibit an ethnic difference.

Objective To investigate the association between CYP1A1 gene polymorphism and toxicities related to high-does methotrexate of childhood acute leukemia. Methods The SNPs were detected by reverse transcriptional (RT)-PCR-denaturing gradient gel elelctrphoresis combined with direct sequencing in 51 children with acute leukemia. Toxicities were collected thereby. Results Only one SNP,A4889G, was screened in CYP1A1. A4889G polymorphism was not associated with all the toxicities (P > 0.05). High-risk ALL children were more likely to increase the risk of thrombocytopenia compared with standard-risk ALL (P< 0.05). Conclusions CYP1A1 A4889G polymorphism may be not association with all toxicities after HD-MTX ,but the thrombocytopenia may be relevant to the risk degree of ALL children.

Objective To investigate the effects of captopril on cardiac function and levels of energy-rich phosphates in pressure overload induced left ventricular hypertrophy rats. Methods One hundred and twenty SD rats were randomly divided into three groups: sham operation group (SH group, n=40),coarctation of abdominal aorta group (CAA group, n=40) and captopril treatment 1 mg· 1001·d-1) group (CAP group, n=40). Left ventricular end-diastolic pressure (LVEDP), left ventricular mass index (LVMI), levels of energy-rich phosphates and morphological changes of the myocardial mitochondria were compared at the 6th and 8th week after operation. Results At 6th week, in CAA group, LVMI and LVEDP were increased and ±dp/dtmax was decreased, while ATP and ADP were decreased and AMP was increased (P<0.01). These changes were much obvious at 8th week (P<0.01). Compared with those of CAA group, the parameters of heart function and energy-rich phosphates (ATP, ADP, AMP, TAN) in CAP group were improved significantly(P<0.01) at the 6th and 8th week. In CAP group, the parameters of heart function and energy-rich phosphates (ADP, AMP, TAN) were much better at 8th week than those at 6th week. The morphological change of mitochondria was less in CAP group than that in CAA group. Conclusion Captopril significantly improves myocardial energy metabolism in pressure overload rats and protects the function of myocardial mitochondria.

Background and purpose: Previous studies have confirmed that the expression of leucine-rich repeat-containing 3B (CLRRC3B) was significantly decreased in different human cancers, which was also associated with the migration and invasion of cancer cells. The aim of this study was to explore the potential mechanism of LRRC3B in the development of esophageal cancer. Methods: The LRRC3B expression was detected in 60 cancer tissues and 60 adjacent non-neoplastic tissues by immunohistochemistry. The mRNA and protein expression of LRRC3B in Eca109 and HEECs were detected using real-time fluorescence quantitative polymerase chain reaction (RTFQ-PCR) and Western blot, respectively. Eca109 cells with different treatments were divided into three groups:normal group, negative control group (transfected with pCMV6 plasmid), overexpression LRRC3B group (transfected with pCMV6-LRRC3B plasmid). Transwell assay was used to measure the migration and invasion of Eca109 cells in different groups. The protein levels of E-cadherin, N-cadherin, Vimentin and p-Akt were determined by Western blot. Results: The expression of LRRC3B in esophageal cancer tissues was lower than that of non-cancerous tissues, as well as the expression of LRRC3B in Eca109 was decreased compared with that of normal esophageal epithelial cell line HEEC. Overexpression of LRRC3B significantly inhibited Eca109 cells migration and invasion, upregulated the expression of E-cadherin and decreased the expression of N-cadherin and Vimentin. Moreover, overexpression of LRRC3B significantly inhibited the phosphorylation of Akt in Eca109 cells. Conclusion: The expression of LRRC3B was decreased in esophageal cancer. Overexpression of LRRC3B can efficiently inhibit the EMT progression in esophageal cancer cells by suppressing PI3K/Akt signaling pathway.

Objective To investigate the effects of CNTN-1 on the invasion and migration of human esophageal cancer EC9706 cells and the possible mechanism.Methods The expression of CNTN-1 in human esophageal cancer EC9706 cells was measured by qPCR and Western blot.After transfection with CNTN-1 siRNA or CNTN-1, the cells were divided into control group, scrambled siRNA group, CNTN-1 siRNA group, pcDNA3.1-vector group and pcDNA3.1-CNTN-1 group.Cell proliferation, invasion and migration were respectively analyzed by BrdU assay and Transwell test.The expression of MMP-2 and MMP-9 were detected by qPCR and Western blot.Results The mRNA and protein expression of CNTN-1 were significantly upregulated in EC9706 cells.Compared with control, cell proliferation, invasion and migration, as well as the expression of MMP-2 and MMP-9 were significantly decreased by CNTN-1 siRNA, while they were increased by CNTN-1 overexpression (P<0.05).ConclusionsCNTN-1 can influence the invasion and metastasis of esophageal cancer cells through the regulation of the expression of MMP-2 and MMP-9.

Objective:To investigate the correlation between polymorphisms of serine hydroxymethyltransferase1 gene and the adverse reactions of high-dose methotrexate (HD-MTX) in children with acute lymphoblastic leukemia (ALL). Methods:A total of 51 patients with ALL were treated with HD-MTX, and clinical manifestations after HD-MTX treatment were evaluated retrospectively. cD-NA was obtained from mRNA. The polymorphisms of SHMT1 gene containing rs1979277, rs3783, rs1979276, and rs12952556 sites were tested by denaturing gradient gel electrophoresis and direct sequencing. Effects of SHMT1 gene polymorphisms on HD-MTX ad-verse reactions were evaluated. Results:Severe adverse reactions in ALL patients treated with HD-MTX appeared to be mainly neutro-penia and hepatoadverse reactions. The frequency distributions of rs3783 (C>G), rs1979276 (C>T), rs12952556 (A>G), and rs1979277 (C>T) were the same. The polymorphisms of rs1979277 showed no correlation with neutropenia (P>0.05) but rs1979277 CT and TT genotypes were correlated with hepatoadverse reactions (CT: OR=0.129, 95% CI: 0.020 to 0.817, P=0.03; TT: OR=0.103, 95% CI:0.017 to 0.620, P=0.013). Conclusion: No correlation was found between the combination of rs1979277, rs3783, rs1979276, rs12952556, and neutropenia, but one or more of these loci may reduce the risk of hepatoadverse reactions.

Objective:To investigate the association between glutathione S-transferase pi (GSTP1) gene polymorphism and toxici-ties related to high-dose methotrexate (HD-MTX) in children with acute lymphoblastic leukemia (ALL). Methods:GSTP1 genotypes and allelic frequencies in 51 children with ALL were determined by Nest PCR, denaturing gel gradient electrophoresis (DGGE), and DNA sequencing. HD-MTX adverse reactions were analyzed using the National Cancer Institute Common Toxicity Criteria (NCICTC). Results:We identified three SNPs of GSTP1, including rs1695 (A313G), rs1138272 (G439T), and rs4891 (T555C). The wild types, het-erozygous types, and homozygous types of GSTP1 rs1695/rs4891 polymorphisms were detected in 32 cases (62.7%), 16 cases (31.4%), and 3 cases (5.9%), respectively. GSTP1 rs1695/rs4891 polymorphisms included only one heterozygous type and one homozygous type. The allele frequencies of the three SNPs were 21.6%, 2.9%, and 21.6%. The AG+GG/TC+CC genotype of GSTP1 rs1695/rs4891 was associated with decrease in the odds of peripheral hemoglobin (OR=0.25, 95%CI=0.06-1.00, P=0.049). The AG+GG/TC+CC genotype of GSTP1 rs1695/rs4891 in standard and intermediate-risk ALL children was significantly correlated with higher odds of gastrointesti-nal toxicity (OR=0.125, 95%CI=0.02-0.78, P=0.026). Conclusion:GSTP1 rs1695 (A313G)/rs4891 (T555C) gene polymorphism is as-sociated with the reduction of peripheral hemoglobin in ALL children and with the odds of gastrointestinal toxicity in standard and inter-mediate-risk ALL children who receive high-dose methotrexate.

The fabrication and experimental test results were presented for a micro-electro-mechanical systems (MEMS)-based gas chromatography column. Compared with conventional column, this micro-gas chromatography column is suitable for on-line analysis and monitoring because of the small size, rapid analysis and other characteristics. These 0.5, 1 and 3 m GC columns were fabricated using a deep reactive-ion etching (DRIE) and were coated the stationary phase via a dynamic coating procedure. These GC columns perfectly separated the mixture of benzene, toluene and o-xylene in less than 200 s, where, the 3.0 m of the GC column achieved) a high resolution of 14.3 between toluene and o-xylene and yielded approximately 6160 plates. Moreover, the effect of separation performance for different column length were examined and compared. These GC columns are suitable for the application of a variety of disciplines, including environmental analysis, methane gas probes and homeland security.

Objective To extract,isolate and identify the polysaccharide from ginseng,and to investigate its anti-tumor activity in vitro. Methods The ginseng polysaccharide was obtained through water extraction and ethyl alcohol deposition method. Use the Sevage method to remove the protein in the crude polysaccharide. The structural characteristics of the polysaccharide were determined by FT-IR spectra.The RM-1 and HeLa cells were divided into control group and different concentrations (0,50,100,200,300,400 and 500 mg·L-1 )of ginseng polysaccharide groups. The survival rates of the cells in various groups were detected by MTT method. The indirect killing effects of ginseng polysaccharide with different concentrations (0,50,100,200,300,400 and 500 mg·L-1 )on the cancer cells were determined by CTL test. Results The extraction rate of ginseng polysaccharide was 8.7 6% and the structural characteristics demonstrated that the main component of the extract was polysaccharide.The result of MTT showed that there were no significant differences of the survival rates of RM-1 and HeLa cells between different concentrations of ginseng polysaccharide groups after treated for 24 h compared with control group (P>0.05).The result of CTL test showed that the cytototic LHD release rates in different concentrations of ginseng polysaccharide groups were increased significantly (P<0.05)compared with control group;with the increase of ginseng polysaccharide concentration, the cytotoxic LDH release rates were increased firstly and then were decreased.When the concentration of ginseng polysaccharide was 100 mg·L-1 ,the cytotoxic LDH release rate was the biggest.Conclusion Ginseng polysaccharide can indirectly inhibit the growth of the tumor cells by activating T cells and play an anti-tumor effect.

Objective To extract the Ginseng polysaccharide (GPS), polysaccharides of Tricholoma matsutake (PTM)and polysaccharide of Lentinus edodes (PLE)from gingeng, tricholoma matsutake and lentinus edodes respectively,and to analyze and identify their structures,and to prepare their complex,and to study the indirect antitumor activity invitro of polysaccharide complex.Methods The polysaccharides were extracted with hot water and precipitated by ethanol.The carbohydrate levels were determined by the method of phenol-sulfuric acid.The m-hydroxyphenyl method was used to determine the levels of uronic acid, and the national standard method was used to determine the levels of starch.Infrared spectroscope and chemical methods were performed to analyze their structures. Orthogonal experiment was used to study mixing methods. Cytotoxic T lymphocyte experiment and LDH release assay were performed to detect the influence of polysaccharide complex of GPS,PTM,and PLE in the CTL killing activity,and its indirect killing effect on the P815 cells.Results The extraction rates of GPS,PTM, and PLE were 8.85%,9.40%,and 10.50%;the levels of total polysaccharides were 62.96%,59.13%,and 33.86%;the levels of uronic acid were 16.44%,9.37%,and 16.44%;the starch levels were 7.26%,2.80%,and 3.77%,respectively.The identification results showed that the polysaccharides were obstrained.When the quality ratio of the three kinds of polysaccharides was 1∶1∶1 and the concentration was 600 mg·L-1 ,the CTL cytotoxicity was the highest.Conclusion The polysaccharide complex is obtained,identified and characterized. Polysaccharide complex can enhance the cytotoxicity of CTL and has the indirectly inhibitory effect on the proliferation of P815 cells.