Molecular cloning and characterization of two genes encoding peroxiredoxins from freshwater bivalve Anodonta woodiana: Antioxidative effect and immune defense.

Members of Prx family function as an important players in host defense against oxidative stress, and modulate immune responses. In the current study, two complete Prx sequences were isolated from bivalve Anodonta woodiana and respectively named AwPrx4a and AwPrx4b. Regulative characterizations of AwPrx4a and AwPrx4b derived from perfluorooctanesulfonate (PFOS), perfluoroocanoic acid (PFOA), lipopolysaccharide (LPS) and polyinosinic:polycytidylic (Poly I:C) challenge in hepatopancreas, gill and hemocytes were measured by quantitative real-time PCR, respectively. The full-length cDNA of AwPrx4a had an open reading frame ORF of 588 bp encoding 196 amino acids. Two highly conserved Prxs signature motifs were observed in deduced amino acid sequence, one was FYPLDFTFACPTEI, and the other was GEVCPA. Complete cDNA sequence of AwPrx4b was comprised of a 5' untranslated region (UTR) of 120 nucleotides, a 426 bp ORF which was encoded 142 amino acids, and a long 3'-UTR of 412 nucleotides. Expressions of AwPrx4a and AwPrx4b showed a significant up-regulation pattern in groups at lower concentration treatment of PFOS and PFOA, a biphasic profile in groups with a higher concentration treatment. Compared with that of control group, expressions of AwPrx4a and AwPrx4b were significantly induced by LPS and Poly I:C treatment in the hepatopancreas, gill and hemocytes. These results indicate up-regulations of AwPrx4a and AwPrx4b expression are associated with eliminating oxidative stress derived from PFOS and PFOA administration as well as enhancing immune defense against LPS and Poly I:C challenge.

Mitochondrial DNA haplogroup D4b is a protective factor for ischemic stroke in Chinese Han population.

Mitochondrial DNA (mtDNA) haplogroups affect the assembly and stability of the mitochondrial respiratory chain, which is potentially related to susceptibility to ischemic stroke (IS). However, the role of mtDNA in IS has not been comprehensively studied. The purpose of this study was to explore whether mtDNA polymorphisms and haplogroups are involved in the etiology of IS in the Chinese Han population. We recruited 200 patients with IS and 200 matched controls and genotyped them for 18 mtDNA single nucleotide polymorphisms defining the major Eastern Asian haplogroups by SNaPshot minisequencing. We also sequenced the hypervariable segment I (HVS-I), position 16051-16400. The prevalence of haplogroup D4b was significantly lower in IS patients than in healthy controls (0 and 8 %, respectively, corrected P = 2 × 10(-5), odds ratio = 0.028, 95 % confidence interval = 0.002-0.468).The positive association between haplogroup D4b and IS may be related to the protective effect of haplogroup D4b against oxidative damage, which decreases the risk of IS. Our study provides the first evidence that haplogroup D4b is a potential genetic protective factor for IS in the Chinese Han population.

Prediction of the potential suitable habitat of Echinococcus granulosus, the pathogen of echinococcosis, in the Tibetan Plateau under future climate scenarios.

Echinococcosis is a zoonotic parasitic infectious disease caused by human or domestic animals infected with Echinococcus granulosus. China is the country with the heaviest disease burden caused by Echinococcosis in the world. Therefore, it is feasible to evaluate the prevalence and distribution of echinococcosis using relevant ecological methods, combined with environmental factors and human activities. In this study, MaxEnt was used to predict the distribution range of E. granulosus in China under current and future climate scenarios and explain the impact of environmental variables on its distribution. The results showed that elevation (El), annual mean temperature (bio1), human footprint (Hf), annual precipitation (bio12), mean temperature of warmest quarter (bio10), and mean temperature of wettest quarter (bio8) were identified as the dominant environmental variables. In Tibet, the most suitable habitats (25.9 × 104 km2) of E. granulosus were distributed in Nyingchi and Qamdo in the east, Shigatse and Shannan in the south, and Ali in the west. In Sichuan, the most suitable habitat (18.83 × 104 km2) was located in Aba, Ganzi, and Liangshan. In Qinghai, the most suitable habitat (13.05 × 104 km2) mainly included Yushu in the southwest; Guoluo in the southeast; Haidong, Huangnan, Xining, and Hainan in the east; and Haixi in the west. In Gansu, the most suitable habitat (7.36 × 104 km2) was located in Gannan and Linxia in the southwest and Wuwei and Dingxi in the middle. In Yunnan, the most suitable habitat (1.53 × 104 km2) was distributed in Diqing in the northwest. Under future climate scenarios, the area of the most suitable habitat of E. granulosus showed an obvious expansion trend, with an increase of 44.64-70.76%. Trajectory trend of centroids showed that the most suitable habitat would move to the west in the future, and the increased areas were mainly located in the west of the current most suitable habitat. AUC values of the training data and test data were 0.936 ± 0.001 ~ 0.97 ± 0.006 and 0.912 ± 0.006 ~ 0.956 ± 0.015, respectively. The result can provide a theoretical basis for the prevention, monitoring, and early warning of echinococcosis in China.

Polymorphisms of the Matrix Metalloproteinase Genes are Associated with Acute Ischemic Stroke in Chinese Han Population.

Background and Purpose: Studies have shown that matrix metalloproteinase (MMP-2,3,9) plays an important role in the pathologic process of ischemic stroke (IS). The aim of this study was to investigate the relationship between C1306T, 1612-5A/6A, C-1562T polymorphisms of MMP-2,3,9 genes and IS in Chinese Han population. Methods: The polymorphisms of MMP-2(C1306T), -3(1612-5A/6A), -9(C-1562T) gene were detected by PCR-RFLP and SNaPshot sequencing. Then, stratified analysis was used to study the relationship between IS subtypes and MMP-2,3,9 polymorphisms. Results: For the MMP-2 gene C1306T polymorphism, TT genotype and T allele were significantly associated with a reduced risk of IS (P = 0.015, P = 0.003, respectively). T allele was significantly associated with a reduced risk of small artery occlusion (SAO) subtype compared with the control group (P = 0.012, OR = 0.550, 95% CI = 0.065-1.291). For the MMP-3 gene-1612 (5A/6A) polymorphism, 5A/5A genotype was significantly increased in the IS group (P = 0.011, OR = 0.370, 95% CI = 0.168-0.814), especially in the large-artery atherosclerosis (LAA) subtype (P = 0.001, OR = 2.345) as compared to the control group. Conclusion: Our study suggested that the T allele of MMP-2 may be a protective factor of IS, especially in SAO subtype, while the 5A/5A gene of MMP-3 may increase the risk of IS, especially in LAA subtype in Chinese Han population.

[Genetic polymorphisms of six short tandem repeat loci in the Han population of Henan province in China].

OBJECTIVE: To investigate the allele frequencies of six short tandem repeats (STR) loci D12S391, D5S818, D18S51, PAHI3, D8S1179, D3S1358 in the Han population of Henan province and to obtain preliminary data. METHODS: DNA was extracted with phenol-chloroform from 140 EDTA-blood specimens of healthy unrelated individuals in Henan population; multiplex PCR technique and PAGE vertical electrophoresis were used to screen the genotype frequencies of six STR systems in Henan population. RESULTS: The test for Hardy-Weinberg equilibrium revealed that the genotype distribution was correspondent with the expected. The observed heterozygosities of six loci were 0.871, 0.769, 0.871, 0.773, 0.901, 0.722. The calculated discrimination power is 0.9999998, the calculated power of exclusion is 0.99845, the calculated matching probability is 2.39 x 10(-7). CONCLUSION: All of the six loci in this study have high power of discrimination and exclusion; they may be very useful genetic markers for individual identification, paternity test and genetics purposes.

Identification of osteosarcoma-related specific proteins in serum samples using surface-enhanced laser desorption/ionization-time-of-flight mass spectrometry.

Osteosarcoma (OS) is the most common malignant bone tumor. To identify OS-related specific proteins for early diagnosis of OS, a novel approach, surface-enhanced laser desorption/ionization-time-of-flight mass spectrometry (SELDI-TOF-MS) to serum samples from 25 OS patients, 16 osteochondroma, and 26 age-matched normal human volunteers as controls, was performed. Two proteins showed a significantly different expression in OS serum samples from control groups. Proteomic profiles and external leave-one-out cross-validation analysis showed that the correct rate of allocation, the sensitivity, and the specificity of diagnosis were 100%. These two proteins were further identified by searching the EPO-KB database, and one of the proteins identified as Serine rich region profile is involved in various cellular signaling cascades and tumor genesis. The presence of these two proteins in OS patients but absence from premalignant and normal human controls implied that they can be potential biomarkers for early diagnosis of OS.

A novel risk haplotype of ALOX5AP gene is associated with ischemic stroke in Chinese Han population.

Previous studies have implicated that two at-risk haplotypes (HapA and HapB) of gene-encoding 5-lipoxygenase-activating protein (ALOX5AP) were significantly associated with stroke. The aim of this study was to explore the association between haplotypes of ALOX5AP gene and risk for ischemic stroke (IS) in Chinese Han population. A total of 492 patients with IS and 490 matched control subjects were recruited. Six ALOX5AP SNPs (SG13S377, SG13S114, SG13S41, SG13S89, SG13S32 and SG13S35) were genotyped by SNaPshot minisequence technique. A common genetic variant SG13S114/AA in the ALOX5AP gene was associated with IS in this Chinese cohort (OR = 2.514, 95 % CI = 1.667 ~ 3.790). HapA (TGA) and HapB (AAAG) had no significant difference in the patients (36.3 and 18.5 %, respectively) and controls (37.6 and 16.3 %, respectively) (P = 0.631 and P = 0.375, respectively). But, the frequency of Hap (GAAG) was significantly higher in the patients than that in the controls after Bonferroni's adjustment (P = 0.006). To conclude, SG13S114/AA of the ALOX5AP gene was associated with an increased risk for IS. A novel risk haplotype, Hap (GAAG) was a genetic risk factor for IS in this Chinese population.

Sex-dependent association of phosphodiesterase 4D gene polymorphisms with ischemic stroke in Henan Han population.

BACKGROUND: Recent evidence has implicated the gene for phosphodiesterase 4D (PDE4D) as susceptibility gene for ischemic stroke (IS) in Icelandic population. However, there are few reports on the associations between PDE4D gene polymorphisms and IS in Chinese individuals. The present study aimed to investigate the possible association of genetic polymorphisms in PDE4D gene with IS in Henan Han population. METHODS: A total of 400 patients with IS and 400 matched controls were examined using a case-control design. Two single nucleotide polymorphism (SNPs) (rs918592 and rs2910829) in PDE4D gene were genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. Odds ratios (OR) and 95% confidence intervals (95%CI) were calculated to test the association between the genetic factors and IS. Genetic parameter and association studies were carried out with SPSS 16.0. RESULTS: Among the two SNPs tested, the rs918592 was significantly associated with IS (OR: 1.351, 95%CI: 1.110 - 1.645), especially in male patients (OR: 1.427, 95%CI: 1.105 - 1.844). Haplotype analysis showed that A-T was associated with an increased risk of the IS (OR: 2.114, 95%CI: 2.005 - 2.230) while G-T was associated with decreased risk of IS (OR: 0.419, 95%CI: 0.302 - 0.583). Protecting effect of haplotype G-T was also significant in males (OR: 0.264, 95%CI: 0.162 - 0.431). CONCLUSIONS: The present study demonstrated a strong association of rs918592 with IS. Haplotype A-T increased the risk of IS while haplotype G-T had a protective effect in Henan Han population. The association was sex-dependent with male patients showing stronger effect.

[Identification of colorectal cancer using proteomic patterns in serum].

BACKGROUND & OBJECTIVE: At present there is no serological parameter with high sensitivity and specificity to diagnose colorectal cancer. This study aimed at establishing a serum protein fingerprinting technique coupled with a pattern-matching algorithm to distinguish colorectal cancer from benign colorectal diseases and healthy people. METHODS: Preliminary group and test group were both random selected serum samples, which each comprises 73 cases of colorectal cancer, 31 healthy people, and 16 cases of benign colorectal diseases. The sera of the test group was combined with the surface of the IMAC3 proteinchip. Then reading data of surface enhanced laser desorption/ionization- time of flight-mass spectrometry (SELDI-TOF-MS) was analyzed by Biomarker Wizard software and Biomarker pattern software to get a classification rule of tree, which is standard configuration that can distinguish the sera of colorectal cancer patients from the sera of benign colorectal disease patients and healthy people, and the standard was approved test validity by double-blindly in the test group. RESULTS: At the key M/Z values of 4 467 Da, 8 131 Da, 8 939 Da, 9 192 Da, 9 134 Da, 8 221 Da, 5 928 Da, 8 324 Da, 11 732 Da, protein contents of the three classes in preliminary group are obviously different by the software analysis. And the classification rule of tree was discovered, and corresponding correct ratio was 98.33% (118/120), corresponding sensitivity was 97.26% (71/73) and corresponding specificity was 100% (47/47); after double-blind examining the test group with that rule, the corresponding correct ratio was 96.77% (116/120),the corresponding sensitivity was 95.89% (70/73) and the corresponding specificity was 97.87%(46/47). CONCLUSION: Via comparative proteomics research in the sera of colorectal cancer patients and benign colorectal disease patients and healthy people, the group of protein was obviously different in pathological process in discovering colorectal cancer.