Mitochondrial DNA Copy Number Variations and Serum Pepsinogen Levels for Risk Assessment in Gastric Cancer

Background:Variations in mtDNA-CN of PBLs, as a potential biomarker for GC screening has currently been subject to controversy. Herein, we have assessed its efficiency in GC screening, in parallel and in combination with sPG I/II ratio, as an established indicator of gastric atrophy. Methods:The study population included GC (n = 53) and non-GC (n = 207) dyspeptic patients. The non-GC group was histologically categorized into CG (n = 104) and NM (n = 103) subgroups. The MtDNA-CN of PBLs was measured by quantitative real-time PCR. The sPG I and II levels and anti-H. pylori serum IgG were measured by ELISA. Results:The mtDNA-CN was found significantly higher in GC vs. non-GC (OR = 3.0; 95% CI = 1.4, 6.4) subjects. Conversely, GC patients had significantly lower sPG I/II ratio than the non-GC (OR = 3.2; CI = 1.4, 7.2) subjects. The combination of these two biomarkers yielded a dramatic amplification of the odds of GC risk in double-positive (high mtDNA-CN-low sPGI/II) subjects, in reference to double-negatives (low mtDNA-CN-high sPGI/II), when assessed against non-GC (OR = 27.1; CI = 5.0, 147.3), CG (OR = 13.1; CI = 2.4, 72.6), or NM (OR = 49.5; CI = 7.9, 311.6) groups. Conclusion:The combination of these two biomarkers, namely mtDNA-CN in PBLs and serum PG I/II ratio, drastically enhanced the efficiency of GC risk assessment, which calls for further validations. Key Words: Biomarkers, DNA copy number variation, Mitochondrial DNA, Stomach neoplasms     

Impact of Methylenetetrahydrofolate Reductase C677T Polymorphism on the Risk of Gastric Cancer and Its Interaction with Helicobacter pylori Infection

Background

Attempts for early detection of gastric cancer have recently focused on host''s genetic susceptibility factors and gene-environment interactions. We have, herein, studied the association of MTHFR C677T single nucleotide polymorphism (SNP) and its interaction with Helicobacter pylori infection, smoking, age and gender on the risk of gastric cancer among an Iranian population.

Methods

Gastric cancer patients (n = 450) and cancer-free controls (n = 780) were studied for serum H. pylori-specific IgG antibodies by ELISA and MTHFR C677T polymorphism (SNP) by PCR-RFLP. Demographic and life style data were collected through patient interviews. Unconditional logistic regression model estimated odds ratio (OR) and the corresponding 95% confidence intervals (CI).

Results

The interactions of MTHFR genotype with H. pylori infection (P = 0.03), age (P = 0.049) and gender (P = 0.007) were statistically significant. Accordingly, MTHFR C677T carriers who were also positive for H. pylori infection exhibited 80% (OR = 1.8, 95% CI = 1.0-2.9) significant excess risk of non-cardia gastric cancer. Furthermore, subjects over the age of 50 or female subjects carrying MTHFR C677T SNP showed 40 (OR = 1.4, 95% CI = 1.0-2.0) and 100 (OR = 2.0, 95% CI = 1.2-3.2) percent increased risk of gastric cancer, respectively.

Conclusion

MTHFR C677T SNP seems to increase the risk of gastric cancer and the effect is significantly inflated by interactions with H. pylori infection, age and gender. Key Words: Helicobacter pylori, Smoking, Gender identity, Age group, Methylenetetrahydrofolate reductase     

Interaction between LINC-ROR and Stemness State in Gastric Cancer Cells with Helicobacter pylori Infection

Background:LINC-ROR, as a cancer-related lncRNA, has vital roles in stem cell survival, pluripotency, differentiation, and self-renewal in hESCs. However, cancer-related molecular mechanisms, its functional roles, and clinical value of LINC-ROR in GC remain unclear. In this study, we aimed to investigate probable interplay between LINC-ROR with SALL4 stemness regulator and their role with the development of the disease. Methods:The mRNA expression profile of LINC-ROR and SALL4 was assessed in tumoral and adjacent non-cancerous tissues of GC patients, using quantitative real-time PCR. Results:Significant LINC-ROR underexpression and SALL4 overexpression were observed in 55.81% and 75.58% (p < 0.0001) of samples, respectively. The expression of LINC-ROR and SALL4 were significantly correlated with each other (p = 0.044). There was an association between the underexpression of LINC-ROR and sex, stage of tumor progression, tumor type, and location of tumor (p < 0.05), and H. pylori infection with SALL4 expression (p = 0.036). There were also significant correlations between concomitant mRNA expression of SALL4 and LINC-ROR in tumors located at distal noncardiac, positive for H. pylori infection, tumors with invasion into the muscle layer of the stomach, and grade II tumor (p < 0.05). Conclusion:The clinical results of the SALL4-LINC-ROR association propose a probable functional interaction between these markers in tumor maintenance and aggressiveness. Our study can help to understand one of the mechanisms involved in the progression of GC through the function of these regulators. Key Words: LINC-ROR, Non-coding RNA, SALL4     

Comparative efficacy sensitivity and specificity of the tests used for the Diagnosis of Helicobacter pylori

Abstract: A total number of 157 samples were examined by 4 different tests-In-house rapid urease (iRUT), Culture, Histopathology and Immunochromatography (Immuno CardSTAT) for the detection of Helicobacter pylori from the patients reported to Hospital Kuala Lumpur, Malaysia during 2007 to 2008. Out of the samples examined 47 (29.9%) were positive for H. pylori by the tests used in the laboratory. Efficacy of detection of the bacteria by the tests- In-house rapid urease, Culture, Histopathology and Immuno CardSTAT were 31.8, 13.9, 30.3 and 32.8%, respectively. However, sensitivity and specificity of the iRUT were 91.5 and 93.6%, respectively and the Positive Predictive Value (PPV) was 86% and the Negative Predictive Value (NPV) was 96.3%. The sensitivity for Immuno CardSTAT rapid test was 100% and the specificity was 79.3%. The PPV was 50% and the NPV was 100%. Convenient methods to the authors were 'In house rapid urease test and Immunochromatography though variability of specificities were observed.     

Comparison between invasive and noninvasive tests in diagnosis of Helicobacter pylori infection

In this study, the invasive and noninvasive diagnotic tests were compared to choose the appropriate test for diagnostice of H. pylori infection. Helicobacter pylori (H. pylori) is a human pathogen that causes chronic gastritis, has a role in gastric and duodenal ulcer, is involved in gastric carcinogenesis and is regarded as a possible important factor in at least a subset of patients with functional dyspepsia. The diagnosis of H. pylori is an essential element in the management of many common gastrointestinal pathologies. The assessment of each routine invasive and noninvasive test is important. We studied a total of 127 outpatients for the detection of H. pylori infection. The presence of H. pylori infection by invasive tests containing the Rapid Urease Test (RUT), histology (Giemsa staining) and culture in 127 patients. Patients who were positive in culture, or two tests from four tests, invasive and noninvasive, were considered to have H. pylori infection. In noninvasive tests, we evaluated anti-H. pylori IgG and anti-CagA antibodies using commercial Enzyme-Linked Immunoassay (ELISA) and Western blot in dyspeptic patients. Eighty five out of the 127 patients were positive for H. pylori. Helicobacter pylori IgG seropositivity and 35 out of the 127 patients were positive for immunoblot. RUT had sensitivity, specifity and accuracy of 96, 80 and 90.5%, respectively; for Elisa these were 85.2, 33 and 70.5%, respectively and for ELISA with immunoblotting they were 65, 45 and 58.8%, respectively. The results of this study suggest that noninvasive tests (ELISA, immunoblotting) have the lowest and RUT with histology have the highest accuracy. These earlier tests can not be used for accurate infection diagnosis.     

Immunosuppressive proteins isolated from spiral and coccoid cytoplasmic solutions of Helicobacter pylori

The aim of this study was to investigate the antiproliferative proteins that probably have a role in Helicobacter pylori evade of immune response and cause chronic infection disease and also to see if coccoid form had a role in its chronicity. H. pylori strain VacA s2/m2 positive and CagA negative, from a gastric biopsy of a patient with peptic ulcer disease, was isolated and cultured in brucella agar. Both spiral and coccoid forms were harvested and ruptured by sonication. The cytoplasmic solutions of both forms were collected and their fractions obtained by gel chromatography and preparative polyacrylamide gel electrophoresis. The fractions were analyzed by MTT assay for their antiproliferative activity. We isolated two proteins with a significant dose dependent antiproliferative activity that analyzed with sodium dodecyl sulfate-polyacrylamide gel electrophoresis, one of them that was urease positive showed two bands with 61 and 27 kDa, which is resumed to urease of H. pylori, another consist of 57 and 63 kDa. Helicobacterpylori secret some proteins like urease that inhibit immune cells proliferation response against its antigens.     

Rapid urease test, touch cytology and histopathologic assessment in determining infection by Helicobacter pylori in outpatient setting

Helicobacterpylori (HP) is a common cause of gastric infection with serious consequences which is detected by different methods. This study aimed at comparing the diagnostic value of Rapid Urease Test (RUT), Touch Cytology (TC) and histopathologic assessment in outpatients setting. In this cross-sectional study, 51 candidates for upper gastrointestinal endoscopy were recruited in Tabriz Imam Khomeini Teaching Centre in a 24 month period of time. Three biopsy specimens were obtained from gastric antrum during endoscopic intervention. The RUT, TC and histopathologic assessment were performed on each biopsy specimen in each patient. Definite infection by HP was considered when at least 2 out of 3 tests indicated presence of infection. Fifty one patients, 29 females and 22 males with a mean age of 40.10 +/- 12.54 (range: 18-72) years enrolled in this study. Infection by HP was definite in 41 cases (80.4%). The infection rates by RUT, TC and histopathologic examination were 82.4, 82.4 and 76.5%, respectively. The sensitivity, specificity and accuracy of RUT, TC and histopathologic assessment were 92.7, 60 and 66.75%; 100, 90 and 98% and 95.1, 100 and 96.1%, respectively. There were significant agreements between outcomes of the three methods in diagnosis of infection by HP. In conclusion, TC was the most sensitive and histopathologic assessment was the most specific method in diagnosis of infection by HP in outpatient setting. The diagnostic value of RUT was rather low in this regard.     

大豆异黄酮摄入对乳腺癌发病风险影响的荟萃分析

通过荟萃分析研究饮食摄入大豆异黄酮对妇女乳腺癌发病风险的影响.以“大豆异黄酮”、“大豆”、“植物雌激素”和“乳腺癌”为主题词及关键词,联合检索1996年1月至2011年12月Medline、Pubmed、Embase、CNKI和万方数据库发表的有关文献.纳入文献限定于队列研究和巢式病例对照研究.使用RevMan软件进行效应值的合并和异质性检验,使用Stma12.0统计软件进行Meta回归,并通过漏斗图和Egger回归计算发表偏倚.研究共纳入16篇文献,包括6159名乳腺癌病例.通过比较最高摄入组和最低摄入组乳腺癌的发病风险,大豆异黄酮能够使绝经后妇女乳腺癌发病风险降低20％(合并RR=0.80,95％ CI:0.71 --0.91;I2=34％,P=0.17),对绝经前妇女乳腺癌发病风险的影响不具有统计学意义;大豆异黄酮能降低亚洲人群乳腺癌发病风险(合并RR=0.82,95％ CI:0.70～0.96;I2 =23％,P=0.26),但是对欧美人群乳腺癌发病风险的影响不具有统计学意义(合并RR=0.98,95％CI:0.90～1.08;12 =33％,P=0.14).表明饮食摄入大豆异黄酮可降低绝经后妇女乳腺癌的发病风险,对于亚洲妇女作用尤其明显.     

氮水平对水稻汕优64和金南风光合特性的影响

 以缺氮、低氮、中氮、高氮、超高氮盆栽土培的汕优64和金南风为材料，测定植株和叶片的生长状况、叶片总N含量、光合速率、光呼吸速率、气孔导度和叶肉导度的变化。结果表明，供试两品种随着施N量的增加，植株的分蘖数增多,株高增加,气孔密度减少, 叶片总N含量、叶绿素含量、光呼吸速率、叶肉导度等均增加。供试两品种的净光合速率变化存在差异，金南风叶片的净光合速率随着施N量的增加而增加,但汕优64在高氮和超高氮的条件下净光合速率反而下降,说明在高氮和超高氮的条件下光氧化环增加的幅度大于光合还原环。     

Identification of Sequence Variation in the Apolipoprotein A2 Gene and Their Relationship with Serum High-Density Lipoprotein Cholesterol Levels

Background:

Apolipoprotein A2 (APOA2) is the second major apolipoprotein of the high-density lipoprotein cholesterol (HDL-C). The study aim was to identify APOA2 gene variation in individuals within two extreme tails of HDL-C levels and its relationship with HDL-C level.

Methods:

This cross-sectional survey was conducted on participants from Tehran Glucose and Lipid Study (TLGS) at Research Institute for Endocrine Sciences, Tehran, Iran from April 2012 to February 2013. In total, 79 individuals with extreme low HDL-C levels (≤5^th percentile for age and gender) and 63 individuals with extreme high HDL-C levels (≥95^th percentile for age and gender) were selected. Variants were identified using DNA amplification and direct sequencing.

Results:

Screen of all exons and the core promoter region of APOA2 gene identified nine single nucleotide substitutions and one microsatellite; five of which were known and four were new variants. Of these nine variants, two were common tag single nucleotide polymorphisms (SNPs) and seven were rare SNPs. Both exonic substitutions were missense mutations and caused an amino acid change. There was a significant association between the new missense mutation (variant Chr.1:16119226, Ala98Pro) and HDL-C level.

Conclusion:

None of two common tag SNPs of rs6413453 and rs5082 contributes to the HDL-C trait in Iranian population, but a new missense mutation in APOA2 in our population has a significant association with HDL-C.     

Investigation of TNF-α and IL-6 Levels in the Sera of Non-Melanoma Skin Cancer Patients

Background:TNF-α and IL-6 are both pleiotropic cytokines playing major roles in cancer-associated cytokine networks. They have previously been investigated for their function in skin malignancies, mostly melanomas, and studies on NMSC patients are relatively rare. In this study, we aimed to assess the associations of serum levels of IL-6 and TNF-α with NMSCs and its clinicopathological features. Methods:This cases-control study was carried out to investigate the serum levels of TNF-α and IL-6 in 70 NMSC patients, in comparison with 30 healthy individuals, by means of flow cytometric bead-based immuneoassay. Results:Serum levels of both TNF-α and IL-6 were significantly higher in NMSC patients (6.470 vs. 4.355 pg/ml; p = 0.0468, respectively), compared to healthy individuals (3.205 vs. 0.000 pg/ml; p = 0.0126, respectively). In the subgroup analysis, SCC patients had higher serum levels of IL-6 compared to healthy individuals (3.445 vs. 0.000 pg/ml; p = 0.0432). No other significant differences were observed in the serum levels of these two cytokines among different clinicopathological subgroups of the patients. Conclusion:The increased levels of TNF-α and IL-6 in NMSC patients can be introduced as an epiphenomenon of a complex cancer-induced cytokine cascade. Key Words: Biomarkers, Cytokines, Interleukin-6, Tumor Necrosis Factor-alpha     

HOTAIR Induces the Downregulation of miR-200 Family Members in Gastric Cancer Cell Lines

Background:Gastric cancer is the fourth most common human malignancy and the second reason for cancer morbidity worldwide. LncRNA HOTAIR has recently emerged as a promoter of metastasis in various cancer types, including GC, through the EMT process. However, the exact mechanism of HOTAIR in promoting EMT is unknown. Aberrant expression of the miR-200 family has been linked to the occurrence and development of various types of malignant tumors. This study investigates the correlation between the HOTAIR and miR-200 family gene expression patterns in GC cell lines. We investigated the miR-200 and HOTAIR due to their common molecular features in the EMT process. Methods:AGS and MKN45 cell lines were transfected with si-HOTAIR, along with a negative control. The effect of HOTAIR knockdown was also analyzed on cell viability and also on the expression of miR-200 family members, including miR-200a, -200b, and -200c, in cell lines using qRT-PCR. Statistical analysis was performed to find the potential correlation between the expression level of HOTAIR and miRs. Results:Our results showed significant increased miR-200 family expression level in transfected AGS and MKN45 GC cells (fold changes > 2; p < 0.001). Moreover, a negative correlation was observed between HOTAIR and miR-200 expression levels in GC cell lines (p < 0.05). Conclusion:Our findings showed a significant association between miR-200 family and HOTAIR expression levels in GC cell lines. Taken together, the HOTAIR-miR-200 axis seems to play a vital role in human GC, suggesting a potential therapeutic target in future GC treatment. Key Words: Gene expression, Long noncoding RNA, HOTAIR, MicroRNAs     

表没食子儿茶素没食子酸酯和维生素C联用对高尿酸血症小鼠血尿酸水平的影响

以KM雄性小鼠为研究对象,酵母膏（7.5 g·kg^-1）和氧嗪酸钾（250 mg·kg^-1）联合给药建立高尿酸血症小鼠模型,探究表没食子儿茶素没食子酸酯（EGCG）和维生素C（Vc）联用对高尿酸血症小鼠血尿酸水平的影响。将小鼠随机分为6组（n=6）：空白组、模型组、别嘌呤醇组（阳性药组）、EGCG组、EGCG联合Vc组和Vc组,连续给药7 d后测定生化指标。结果表明,与模型组相比,EGCG联合Vc组小鼠的血尿酸值（UA）,血尿素氮（BUN）和肌酐（Cr）水平均明显降低（P<0.001）;EGCG与Vc联用明显抑制了肝脏中腺苷脱氨酶（ADA）和黄嘌呤氧化酶（XOD）的活性（P<0.05或P<0.01）,并显著下调了肾脏中葡萄糖转运蛋白9（GLUT9）mRNA的表达（P<0.001）;肾脏切片显示EGCG和Vc联用显著改善高尿酸血症小鼠的肾脏损伤。此外,EGCG与Vc联用对高尿酸血症小鼠的作用效果优于EGCG。     

Expression Analysis of mir-21 and mir-221 in Cancerous Tissues from Iranian Patients with Gastric Cancer

Background:

Early detection is a key to survival for gastric cancer. Molecular markers such as miRNA (microRNA) can have great importance in the early diagnosis of gastric cancer. Expression of miR-21 and miR-221 are deregulated in many types of human cancers. This study aimed to investigate the differences in miRNA expression patterns within the Iranian population.

Methods:

Total RNA was extracted from gastric cancer tissues and adjacent non-cancerous tissues from 32 patients. Expression levels of miR-21 and miR-221 were detected by Real time RT-PCR using a specific primer, with 5s rRNA as the internal reference gene.

Results:

Our data showed that the expression levels of miR-21 and miR-221 in gastric cancer samples were significantly higher than in paired non-cancerous samples (P < 0.05). The receiver operating characteristic (ROC) analyses yielded the area under the curve (AUC) values of 80.30 for miR-21 and 93.30 for miR-221, and combined ROC analysis revealed the highest AUC value of 96.90 in discriminating GC patients from healthy controls.

Conclusion:

It seems that miR-21 and miR-221 expression pattern in Iranian patients with gastric cancer are similar to any other population. Considering the increased expression level of two miRNAs in cancerous tissue compared to normal tissue as well as the area under ROC curve, miR-21 and miR-221 can be used for early detection of gastric cancer. Key Words: MicroRNAs, Tumor markers, Stomach neoplasms     

Effects of Daily Kelp (Laminaria japonica) Intake on Body Composition,Serum Lipid Levels,and Thyroid Hormone Levels in Healthy Japanese Adults: A Randomized,Double-Blind Study

To investigate whether supplementation with iodine-reduced kelp (Laminaria japonica) powder decreases body fat composition in overweight Japanese subjects, a randomized, double-blind, placebo-controlled intervention study was conducted in 50 Japanese subjects with body mass index (BMI) ≥25 and <30 kg/m². Subjects were randomly assigned to consume thirty tablets/d (10 tablets orally, 3 times/d) containing either iodine-reduced kelp powder (test, 6 g kelp powder corresponding to 3 g alginate/d) or kelp-free powder (placebo) for 8 weeks. Anthropometric measurements, blood lipids, and serum thyroid hormone levels were obtained before and after the trial. Body fat percentage was significantly decreased in male subjects from the test group compared with the placebo group. The same tendency was observed for body weight (p = 0.065) and BMI (p = 0.072) in male subjects. No significant changes in anthropometric measurements or visceral fat area were observed in female subjects. Serum thyroid hormone concentrations did not increase after 1.03 mg/d of iodine supplementation through kelp intake. The intake of iodine-reduced kelp powder led to significant and safe reductions in body fat percentage in overweight male subjects. The consumption of kelp high in alginate may contribute to preventing obesity without influencing thyroid function in Japanese subjects with a relatively high intake of iodine from seaweed.     

高光谱与叶绿素计快速测定大麦氮素营养状况研究

为了寻找一种简便、快速、非接触性的方法测定作物氮素营养状况，通过田间试验，利用光谱仪和叶绿素计测量了不同氮素水平及不同时期大麦（秀麦3号、浙农大3号）的冠层光谱及叶片SPAD值，在此基础上分析了大麦冠层光谱特性及一阶导数光谱、红边、叶片SPAD值与氮素水平之间的相关性。结果表明，大麦冠层光谱及其一阶导数光谱和红边与氮素水平存在显著相关，不同供氮水平下叶片的SPAD值具有显著性差异，叶片SPAD值与氮素水平之间的相关系数在孕穗期均达到0.75以上，相关达0.01显著水平，从而说明可以通过光谱来测定大麦的氮素水平。     

甘薯育种亲本及组合评价

对南充市农业科学研究所“十五”期间主要选用的甘薯亲本材料作母本、父本组合的实生苗及其后代表现进行分析,其结果,84-2315、9014-3、AB940078-1、绵粉一号、徐薯18、浙13、三合薯、台农10号是较好的高淀粉育种亲本材料。高淀粉育种应该利用这些亲本多配组合或进行集团杂交,以期从其后代中筛选高淀粉甘薯新品种。84-2315、9014-3、潮薯一号、绵粉一号、徐薯18、南薯99、农林10号、坦桑尼亚是较好的高产饲用型育种亲本材料,高产饲料型品种选育应利用这些亲本材料多配组合或进行集团杂交,以期从其后代中筛选高产饲料型甘薯新品种。     

Risk Assessment and Mitigation of the Mycotoxin Content in Medicinal Plants by the Infusion Process

Plant Foods for Human Nutrition - Medicinal plants are often consumed as infusions with boiled water. Scarce information is available in the literature about the migration of mycotoxins into the...     

基于SDE和HS-SPME/GC-MS的乌龙茶香气组成特征分析

为更好地分析乌龙茶香气组成特征,本试验应用气相色谱-质谱联用（GC-MS）检测平台,分别考察了同时蒸馏萃取法（SDE）和顶空固相微萃取法（HS-SPME,3种不同纤维涂层萃取头）对3个福建代表性乌龙茶样品（铁观音、肉桂和水仙）香气成分的萃取效果。结果表明,采用不同香气成分萃取方法可从供试茶样检出262种香气成分,其中以肉桂香气种类最多（204种）,水仙其次（175种）,铁观音最少（120种）。应用HS-SPME干样萃取法可获得最多种类的检出成分（186种）,其次为HS-SPME湿样萃取法（139种）,并以SDE法检出成分最少（33种）。不同萃取方法对香气成分的检出存在良好的互补作用,但以基于HS-SPME干样萃取法,应用65βμm PDMS/DVB（聚二甲基硅氧烷/二乙烯基苯）萃取头进行检测分析可相对较好地反映3种供试茶样香气化学组成特征,并能实现橙花叔醇、吲哚、α-法呢烯、苯乙醇、苯乙腈、水杨酸甲酯等多种乌龙茶常见或特征香气成分的重复性检测。肉桂和水仙茶样相较铁观音茶样的香气种类和数量存在较大差异,且以肉桂茶样的香气组成更为复杂多样,该结果与闽南乌龙茶与闽北乌龙茶迥然不同的花香品质及肉桂相较水仙拥有更为浓郁高显的香型特征相符合。