Objective To investigate the clinical effects of montelukes in treatement of wheezing infants with viral infections. Methods 90 wheezing infants with viral infections were divided into oral Montelukas treatment( Group A) 50 cases and inhaled corticosteroid treatment (Group B) 40 cases, Group B were treated with conventional therapy plus glucocorticoids, Group A in addition to therapy with Montelukast 4mg/time at a draught quaque nocte for 10 days. The clinical effect 、improvement of symptoms 、 pulmonary function, blood gas analysis and the side effects were observed for the infants in both groups. Results The viral infections-positive with first break in 70 cases (77. 7% ) were significantly higher than that recurrent break in 20 cases (22.2% ) (x2 =6. 65, P ＜ 0.01) ; The scores of PaO2 after treatment were significantly higher than before treatment in two groups( t = 2. 542,2.533,all P ＜ 0. 05 ) ; A total effective rate was 88. 0% in group A which was significantly higher than that 72. 5% in group B( x2 = 3. 751 ,P ＜ 0.05) ;The adverse reactions incidence 4. 0% (2/50) in group A was significantly lower than the B group 10. 0% (4/40) in group B(x2 =3.857,P＜0.05). Conclusion Montelukast had definitely effect for infant wheezing with virus infection, and had less side effects.

Objective To explore the significance of CRP in the diagnosis and treatment of pediatric diseases. Methods The highest CRP value and the total number of white blood cells of hospitalized children during hospitalization were detected ,divided into 6 groups according to the level. To compare the relationship among the total number of WBC,CRP and PCT of each group in the infectious diseases and compare the changes of the total number of WBC and CRP before and after treatment. Results CRP and serum procalcitonin (PCT) was positively correlated (P ＜ 0.01), but a large part of white blood cells did not associate with the PCT. In total white blood cells of 6 groups,3 groups had no statistically significant changes before and after treatment, while the CRP abnormal changes of 5 groups before and after treatment was statistically significant(P ＜0.05). Conclusion CRP was not only as the initial identification of indicators of the children with bacterial infections and viral infections,but also reflected the severity of inflammation. CRP should be used as a routine examination of outpatient with fever, associated with the analysis of the white blood cell could play an important role in reducing the abuse of antibiotics.

Objective To obtain recombinant human pancreatic alpha-amylase protein (AMY2A). Methods Human pancreatic AMY2A cDNA was synthesized by RT-PCR using total RNA from human pancreatic tissues and a couple of primers designed according to the know sequence of human pancreatic alpha-amy lase gene, then digested with BamH Ⅰ and Kpn Ⅰ and inserted into the prokaryotic expression plasmid pGEX-5T vector. Construct The prokaryotic expression vector pGEX-5T-AMY2A was constructed and transformed into E. coli BL21 cell . Protein expressed under the induction of IPTG. The inclusion bodies were isolated and solubilized with urea and washed denatured and refolded. The fusion protein wes purified by affinity chrom atography with glutathione agarose. Results Sequence and restrction analysis revealed AMY2A gene was cloned in frame into pGEX-5T, SDS-PAGE profile showed a clear protein band with a relative molecular weight of 84000 and western blot indicated that the expressed product specifically reacted to polyclonal anti -human pancreatic AMY2A genes. Conclusion Human pancreatic AMY2A gene was successfully cloned,expressed and purification.

Objective To investigate molecular diagnostic method for hereditarymethemoblobinemia. Methods The cDNA coding sequence of NADH-cytochrome b5 reductase (b5R) from 3 patients with hereditary methemoglobinemia was analyzed by direct sequencing of RT-PCR products and the genomic DNA of b5R gene by PCR-restriction endonuclease digestion or PCR-sequencing. Results The b5R cDNA of patient A was T/C heterozygous at nucleotide 527 and G/A heterozygous at nucleotide 608. The b5R cDNA of patient B was G/A heterozygous at both nucleotide 170 and nucleotide 179. The b5R cDNA of patient C was G/A heterozygous at nucleotide 608 and C/T heterozygous at nucleotide 791. Result of genomic DNA analysis was in agreement with that of cDNA approach. Conclusion The method for molecular diagnosis of hereditary methemoglobinemia was established and 3 novel b5R gene mutations were identified in compound heterozygosity in 3 Chinese patients.

OBJECTIVETo investigate the polymorphism in the promoter region of PCA3 gene and its relationship with risk of prostate cancer (PCa).

METHODSThe promoter region of PCA3 gene of the DNA of peripheral blood mononuclear cells was detected by sequence analysis in the 186 PCa and 141 BPH patients and 135 healthy control individuals. If the samples were detected with polymorphism of insection/deletion, clone sequence analysis was used with pBS-T carrier to verify it.

RESULTSThere were 5 polymorphisms. TAAA repeat times: 4, 5, 6, 7, 8, and 8 genotypes (TAAA 4/5, TAAA 4/6, TAAA 5/5, TAAA 5/6, TAAA 5/7, TAAA 5/8, TAAA 6/6, and TAAA 6/7) were detected in the promoter region of PCA3 gene. The eight genotypes were divided into three groups: ≤10TAAA, 11TAAA, ≥12TAAA. Unconditional logistic regression analysis models were used to analyze the relationship between different genotypes and cancer risks adjusted by sex and age. The type 11TAAA and ≥12TAAA was associated with higher relative risk for prostate cancer than the group ≤10TAAA [OR=1.74, 95% CI=1.06-2.87 (for type 11TAAA); OR=5.63, 95% CI=1.85-17.19 (for type ≥12TAAA)]. In the 186 PCa patients, there was 62.4% allele of PCA3 gene with AG/CA mutation found in the promoter 18-19 bp region of PCA3 gene and it had a close relation with the development of prostate cancer.

CONCLUSIONSShort tandem repeats are found in the promoter region of the PCA3 gene in PCa patients, and the increase of TAAA repeat sequences highly enhance the relative risk of prostate cancer development. The occurrence of such STR might be related to the mutations in their upstream loci.

Antigens, Neoplasm ; genetics ; metabolism ; Base Sequence ; Genes, Neoplasm ; physiology ; Genotype ; Humans ; Leukocytes, Mononuclear ; Male ; Microsatellite Repeats ; Mutation ; Polymorphism, Genetic ; Promoter Regions, Genetic ; Prostatic Neoplasms ; epidemiology ; genetics ; Risk

Objective To investigate the polymorphism in the promoter region of PCA3 gene and its relationship with risk of prostate cancer ( PCa) . Methods The promoter region of PCA3 gene of the DNA of peripheral blood mononuclear cells was detected by sequence analysis in the 186 PCa and 141 BPH patients and 135 healthy control individuals. If the samples were detected with polymorphism of insection/deletion, clone sequence analysis was used with pBS?T carrier to verify it. Results There were 5 polymorphisms. TAAA repeat times:4, 5, 6, 7, 8, and 8 genotypes (TAAA 4/5, TAAA 4/6, TAAA 5/5, TAAA 5/6, TAAA 5/7, TAAA 5/8, TAAA 6/6, and TAAA 6/7) were detected in the promoter region of PCA3 gene. The eight genotypes were divided into three groups: ≤10TAAA, 11TAAA, ≥12TAAA. Unconditional logistic regression analysis models were used to analyze the relationship between different genotypes and cancer risks adjusted by sex and age. The type 11TAAA and≥12TAAA was associated with higher relative risk for prostate cancer than the group ≤10TAAA [ OR=1. 74, 95%CI=1. 06?2. 87 ( for type 11TAAA);OR=5. 63, 95%CI=1. 85?17. 19 (for type≥12TAAA)]. In the 186 PCa patients, there was 62. 4% allele of PCA3 gene with AG/CA mutation found in the promoter 18?19 bp region of PCA3 gene and it had a close relation with the development of prostate cancer. Conclusions Short tandem repeats are found in the promoter region of the PCA3 gene in PCa patients, and the increase of TAAA repeat sequences highly enhance the relative risk of prostate cancer development. The occurrence of such STR might be related to the mutations in their upstream loci.

Objective To investigate the polymorphism in the promoter region of PCA3 gene and its relationship with risk of prostate cancer ( PCa) . Methods The promoter region of PCA3 gene of the DNA of peripheral blood mononuclear cells was detected by sequence analysis in the 186 PCa and 141 BPH patients and 135 healthy control individuals. If the samples were detected with polymorphism of insection/deletion, clone sequence analysis was used with pBS?T carrier to verify it. Results There were 5 polymorphisms. TAAA repeat times:4, 5, 6, 7, 8, and 8 genotypes (TAAA 4/5, TAAA 4/6, TAAA 5/5, TAAA 5/6, TAAA 5/7, TAAA 5/8, TAAA 6/6, and TAAA 6/7) were detected in the promoter region of PCA3 gene. The eight genotypes were divided into three groups: ≤10TAAA, 11TAAA, ≥12TAAA. Unconditional logistic regression analysis models were used to analyze the relationship between different genotypes and cancer risks adjusted by sex and age. The type 11TAAA and≥12TAAA was associated with higher relative risk for prostate cancer than the group ≤10TAAA [ OR=1. 74, 95%CI=1. 06?2. 87 ( for type 11TAAA);OR=5. 63, 95%CI=1. 85?17. 19 (for type≥12TAAA)]. In the 186 PCa patients, there was 62. 4% allele of PCA3 gene with AG/CA mutation found in the promoter 18?19 bp region of PCA3 gene and it had a close relation with the development of prostate cancer. Conclusions Short tandem repeats are found in the promoter region of the PCA3 gene in PCa patients, and the increase of TAAA repeat sequences highly enhance the relative risk of prostate cancer development. The occurrence of such STR might be related to the mutations in their upstream loci.

ObjectiveTo investigate the characteristics of mitochondrial translational initiation factor 2 （MTIF2） gene methylation and its association with the development and progression of hepatocellular carcinoma （HCC）. MethodsMethSurv and EWAS Data Hub were used to perform the standardized analysis and the cluster analysis of MTIF2 methylation samples， including survival curve analysis， methylation signature analysis， the association of tumor signaling pathways， and a comparative analysis based on pan-cancer database. The independent-samples t test was used for comparison between two groups； a one-way analysis of variance was used for comparison between multiple groups， and the least significant difference t-test was used for further comparison between two groups. The Cox proportional hazards model was used to perform the univariate and multivariate survival analyses of methylation level at the CpG site. The Kaplan-Meier method was used to investigate the survival differences between the patients with low methylation level and those with high methylation level， and the Log-likelihood ratio method was used for survival difference analysis. ResultsGlobal clustering of MTIF2 methylation showed that there was no significant difference in MTIF2 gene methylation level between different races， ethnicities， BMI levels， and ages. The Kaplan-Meier survival curve analysis showed that the patients with N-Shore hypermethylation of the MTIF2 gene had a significantly better prognosis than those with hypomethylation （hazard ratio ［HR］=0.492， P<0.001）， while there was no significant difference in survival rate between the patients with different CpG island and S-Shore methylation levels （P>0.05）. The methylation profile of the MTIF2 gene based on different ages， sexes， BMI levels， races， ethnicities， and clinical stages showed that the N-Shore and CpG island methylation levels of the MTIF2 gene decreased with the increase in age， and the Caucasian population had significantly lower N-Shore methylation levels of the MTIF2 gene than the Asian population （P<0.05）； the patients with clinical stage Ⅳ had significantly lower N-Shore and CpG island methylation levels of the MTIF2 gene than those with stage Ⅰ/Ⅱ （P<0.05）. Clinical validation showed that the patients with stage Ⅲ/Ⅳ HCC had a significantly lower methylation level of the MTIF2 gene than those with stage Ⅰ/Ⅱ HCC and the normal population （P<0.05）. ConclusionN-Shore hypomethylation of the MTIF2 gene is a risk factor for the development and progression of HCC.

ObjectiveTo investigate the role and mechanism of DNA repair regulation in the process of hepatocellular carcinoma （HCC） recurrence. MethodsHCC tissue samples were collected from the patients with recurrence within two years or the patients with a good prognosis after 5 years， and the Tandem Mass Tag-labeled quantification proteomic study was used to analyze the differentially expressed proteins enriched in the four pathways of DNA replication， mismatch repair， base excision repair， and nucleotide excision repair， and the regulatory pathways and targets that play a key role in the process of HCC recurrence were analyzed to predict the possible regulatory mechanisms. The independent samples t-test was used for comparison of continuous data between two groups； a one-way analysis of variance was used for comparison between multiple groups， and the least significant difference t-test was used for further comparison between two groups. ResultsFor the eukaryotic replication complex pathway， there were significant reductions in the protein expression levels of MCM2 （P=0.018）， MCM3 （P=0.047）， MCM4 （P=0.014）， MCM5 （P=0.008）， MCM6 （P=0.006）， MCM7 （P=0.007）， PCNA （P=0.019）， RFC4 （P=0.002）， RFC5 （P<0.001）， and LIG1 （P=0.042）； for the nucleotide excision repair pathway， there were significant reductions in the protein expression levels of PCNA （P=0.019）， RFC4 （P=0.002）， RFC5 （P<0.001）， and LIG1 （P=0.042）； for the base excision repair pathway， there were significant reductions in the protein expression levels of PCNA （P=0.019） and LIG1 （P=0.042） in the HCC recurrence group； for the mismatch repair pathway， there were significant reductions in the protein expression levels of MSH2 （P=0.026）， MSH6 （P=0.006）， RFC4 （P=0.002）， RFC5 （P<0.001）， PCNA （P=0.019）， and LIG1 （P=0.042） in recurrent HCC tissue. The differentially expressed proteins were involved in the important components of MCM complex， DNA polymerase complex， ligase LIG1， long patch base shear repair complex （long patch BER）， and DNA mismatch repair protein complex. The clinical sample validation analysis of important differentially expressed proteins regulated by DNA repair showed that except for MCM6 with a trend of reduction， the recurrence group also had significant reductions in the relative protein expression levels of MCM5 （P=0.008）， MCM7 （P=0.007）， RCF4 （P=0.002）， RCF5 （P<0.001）， and MSH6 （P=0.006）. ConclusionThere are significant reductions or deletions of multiple complex protein components in the process of DNA repair during HCC recurrence.