Serum and urinary amino acid profiling analysis for urinary stone disease using GC-MS / 中华检验医学杂志

Objective:To analyze the serum and urinary amino acid (AA) profiles of urolithiasis patients to explore the potential biomarkers for clinical screening and early diagnosis.Methods:Case-control study. Serum and urine samples were collected from 74 urolithiasis patients (aged 20-82 years, 41 men, 33 female) in the department of urology of the First Affiliated Hospital of Fujian Medical University and 35 healthy controls (HC, aged 22-80 years old, 20 men, 15 female) from the health examination center from February 2015 to October 2017. Serum and urinary AA levels of patients and HC were analyzed using Gas Chromatography-Mass Spectrometry (GC-MS) based metabolomic strategy. The multivariate statistical analysis methods of principal component analysis (PCA) and orthogonal partial least squares discrimination analysis (OPLS-DA) were employed for modeling. The variable importance projection (VIP) value of OPLS-DA model>1 and P<0.05 of t test were selected to screen the differential amino acid metabolites. The diagnostic capabilities of potential markers were evaluated by receiver operating characteristic (ROC) curve and binary logistic regression analysis. Results:Five AA metabolites including serine, glutamate, aspartic acid, isoleucine and glycine were found, which had statistically significant differences between the patient group and the control group ( P<0.05) and were associated with seven metabolic pathways. Serum serine, glutamate, aspartic acid, isoleucine and urine glycine and aspartic acid were combined into an integrated marker panel whose AUC value was 0.890, the sensitivity was 78.0%, and the specificity was 96.4%. Conclusion:Five amino acids in serum and urine could be used as an integrated biomarker panel for the clinical screening and early diagnosis of urolithiasis, which could provide some experimental basis for molecular urolithiasis research.

Shenmai injection for inhibition of hepatic and renal toxicity and leukocyte disorder during chemoradiotherapy in advanced breast cancer / 中国生化药物杂志

Objective To investigate the Shenmai injection for the inhibition of hepatic and renal toxicity and leukocyte disorder during chemoradiotherapy in the women with advanced breast cancer. Methods 58 cases of female breast cancer patients with stage Ⅳ were selected and randomly divided into 2 groups, 29 cases of each group, and patients were treated with 5-hydroxytryptamine (5-HT3) receptor antagonists and white blood cell growth hormone and other conventional therapy, the control group received gemcitabine plus cisplatin chemotherapy, 28d for 1 cycles, the treatment group received more with Shenmai injection, interval was 15d, 2 groups were treated for 3 cycles. Levels of peripheral blood T lymphocyte subsets, cytokine levels and liver and kidney function, quality of life and clinical efficacy were compared. Results Compared with before treatment, levels of CD3+, CD4+ and CD4+/CD8+ in control group decreased (P<0.05), levels of CD3+, CD4+ and CD4+/CD8+ in treatment group increased (P<0.05), levels of CD8+ decreased(P<0.05), levels of IFN-γ, IL-2 and TNF-α increased(P<0.05), levels of IL-6 decreased(P<0.05), scores of KPS increased(P<0.05), scores of FACT-B decreased(P<0.05), levels of ALT, AST, BUN increased(P<0.05), and levels of CCr, WBC counts decreased(P<0.05), and compared with the control group, levels of CD3+ , CD4+ and CD4+/CD8+ in the treatment group were higher(P<0.05), levels of CD8+ were lower(P<0.05), levels of IFN-γ, IL-2 and TNF-α were higher(P<0.05), levels of IL-6 were lower(P<0.05), and the total efficiency was higher(P<0.05), levels of ALT, AST, BUN were lower (P<0.05), and levels of CCr, WBC counts were higher (P<0.05). After treatment, the efficacy of treatment group was higher than that of control group(Z=-2.142,P=0.032<0.05). Conclusion Shenmai injection can improve the efficacy of radiotherapy and chemotherapy in patients with advanced breast cancer, and it can effectively inhibit the liver and kidney damage and leukocyte disorder.

The association between ulcerative colitis and TRAIL receptor genetic polymorphisms / 中华检验医学杂志

Objective To investigate associations of UC with the polymorphisms of TRAIL receptors.Methods From January 2008 to December 2012, 380 consecutive UC patients [215 males and 165 females, the average age was (42.63 ±14.61) years] as well as 539 sex-and age-matched healthy individuals [290 males and 249 females, the average age was (41.29 ±15.86) years] were recruited from four large scale comprehensive hospitals in Wenzhou city.Five single nucleotide polymorphisms of DR4 (rs20575, rs13278062), DR5(rs1047266), DcR2(rs1133782) and OPG (rs3102735) were detected by a SNaPshot technique.Distributions of mutant alleles and genotypes for targeted polymorphisms in TRAIL receptors were analyzed by Chi-square test or Fisher′s exact test. By means of unconditional Logistic regression analysis, it evaluated associations between the polymorphisms and the risk of UC attack as well as the clinical features of UC patients.Furthermore, an unconditional Logistic multiple regression analysis was employed to investigate the independent risk factors of UC and their multiplicative interaction effects on UC.Results The frequencies of mutant allele (G) and genotype (CG+GG) of DR4(rs20575) were higher in UC patients than those in the controls (3.55%vs 1.95%,χ2 =4.512, P=0.034;6.58%vs 3.71%,χ2=3.938, P=0.047, respectively).However, the frequeucies of mutant allele ( A) and genotype ( GA+AA) of DcR2(rs1133782) were decreased in UC patients compared to the controls(6.18%vs 9.09%,χ2=5.183, P=0.023; 11.32% vs 17.44%, χ2 =6.589, P=0.010, respectively).The frequencies of mutant allele (T) and homozygote (TT) of OPG(rs3102735) were significantly higher in UC patients than in the controls (86.32% vs 81.54%, χ2 =7.385, P=0.007;75.26% vs 66.98%, χ2 =7.346, P=0.007, respectively) .Furthermore, the genotype (GG) of DcR2 (rs1133782) was found to be the independent risk factor for UC attack (OR=4.937, 95%CI:2.320-10.504, P<0.001).Moreover, the (GG) of DcR2(rs1133782) and (CC) of DR4(rs20575) had an interactive effect on UC (OR=0.322, 95%CI:0.164-0.633, P=0.001).The same conclusion was drawn for the ( GG) of DR4( rs20575) and (TT) of OPG(rs3102735) (OR=1.580, 95%CI:1.165-2.144, P=0.003).Conclusions The genetic polymorphisms of DR4 ( rs20575 ) , DcR2 ( rs1133782 ) and OPG ( rs3102735 ) were associated with UC. The mutation of DcR2(rs1133782) might play a protective role in UC.Moreover, the DcR2(rs1133782) and DR4(rs20575) gene had a collaborative effect on UC.So did the DR4(rs20575) and OPG(rs3102735) genes.

Associations of Crohn's disease with DR4 and DR5 gene polymorphisms / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To assess the associations of death receptor DR4 and DR5 gene polymorphisms with Crohn's disease (CD).</p><p><b>METHODS</b>A total of 295 CD patients and 490 healthy controls were recruited. Three single nucleotide polymorphisms (SNPs) of the DR4 (rs13278062, rs20575) and DR5 (rs1047266) genes were determined with a SNaPshot method. Unconditional logistic regression analysis was carried out for determining the allelic and genotypic differences of the three SNPs between CD patients and the controls, as well as the influence of the DR4 and DR5 gene polymorphisms on the clinical features of CD patients. Linkage disequilibrium and haplotype analysis were calculated by haplotype 4.2 and R language software. A gene-gene interaction model was established to analyze whether the three SNPs can exert a synergistic effect on the susceptibility to CD.</p><p><b>RESULTS</b>The mutant allele (T) and genotype (GT+TT) of DR4 (rs13278062) were increased among CD patients compared to the controls (37.12% vs. 32.04%, P = 0.040, 95%CI: 1.010-1.550; 62.71% vs. 54.90%, P = 0.032, 95%CI: 1.028-1.855, respectively). However, the allelic and genotypic frequencies of DR4 (rs20575) and DR5 (rs1047266) did not differ between the two groups (all P > 0.05). Based on the Montreal Classification Standards, the CD patients were stratified by locations and behaviors of the disease. After multiple comparison correction (P < 0.0125), compared to ileocolonic CD patients respectively, the mutant allele (T) and genotype (GT+TT) of the rs13278062 polymorphism were significantly increased in colonic CD patients (41.04% vs. 25.64%, P = 0.002, 95%CI: 0.315-0.778; 66.04% vs. 41.03%, P = 0.001, 95%CI: 0.196-0.655, respectively) and terminal ileum CD patients (41.44% vs. 25.64%, P = 0.002, 95%CI: 0.311-0.762; 74.77% vs. 41.03%, P < 0.001, 95%CI: 0.126-0.437, respectively). In comparison to penetrating CD patients, the mutant allele (T) and genotype (GT+TT) of DR4 (rs13278062) were significantly decreased in stricturing CD patients (32.29% vs. 48.91%, P = 0.007, 95%CI: 0.300-0.828; 57.29% vs. 86.96%, P = 0.001, 95%CI: 0.078-0.520, respectively). A similar conclusion was drawn for the mutant genotype (GT+TT) of DR4 (rs13278062) in non-stricturing, non-penetrating CD patients (58.82% vs. 86.96%, P = 0.001, 95%CI: 0.086-0.536). Haplotype analysis indicated that the CT haplotype formed by rs20575 and rs13278062 was increased in CD patients compared to the controls (37.1% vs. 31.8%, P = 0.029, OR=1.279, 95%CI: 1.022-1.600). The outcome of a gene-gene interaction model indicated that the mutant genotype (GT+TT) of DR4 (rs13278062) and mutant genotype (CT+TT) of DR5 (rs1047266) may play a negatively synergistic role in CD patients (B = - 0.483, OR = 0.617, P = 0.030).</p><p><b>CONCLUSION</b>The rs13278062 polymorphism of the DR4 gene not only can confer an increased risk for CD, but may also influence the location of the lesions and the disease behaviors. The CT haplotype formed by rs20575 and rs13278062 may be an independent risk factor for CD. Furthermore, the mutant genotype (GT+TT) of DR4 (rs13278062) and mutant genotype (CT+TT) of DR5 (rs1047266) may exert a negative synergistic effect on CD.</p>

Determination of Adenovirus Type 7 by Cells Culture Combined with RT-PCR / 中华医院感染学杂志

OBJECTIVE To establish a method for detecting adenovirus type 7 by cells culture combined with real-time fluorescent RT-PCR.METHODS After purified adenovirus was dissociated from nasopharyngeal secretion in A549 cells,ADV7 E1A genes were detected by real-time RT-PCR assay and sequence analysis of cells infected with 0.1,0.5,5.0 and 10.0 MOI ADV7 at 3,6,12 and 24 h postinfection.Then the adenovirus in nasopharyngeal secretion was detected with the similar method.RESULTS Early transcription of E1A genes of adenovirus type 7 could be detected by real-time RT-PCR at 3 h postinfection with 0.5MOI virus;or at 6 h postinfection with 0.1MOI virus;Early transcription of E1A genes could be detected at 6 h postinfection in nasopharyngeal secretion.CONCLUSIONS The method by cells culture combined with real-time fluorescent RT-PCR is sensitive,specific and rapid.It can be applied in clinics for diagnosis of adenovirus type 7 infection.