Intratumoral heterogeneity of CD44v6 in rectal cancer.

PURPOSE: CD44v6 plays a controversial role in tumor progression and patient outcome in colorectal cancer by plenty of conflicting reports. The purpose of this study was to profile the intratumoral heterogeneity of CD44v6 in rectal cancer and investigate its role in lymph node metastasis. METHODS: Sixty patients were included in this study. Immunohistochemistry for CD44v6 was performed in normal mucosa, primary tumor, and lymph node metastasis with whole tissue sections. The staining intensity in tumor center and invasive front was separately measured. Sampling bias was evaluated by quantitative real-time PCR with 15 pairs of frozen tissues from different sites of the primary tumor. RESULTS: CD44v6 expression increased from normal mucosa to primary tumor to lymph node metastasis. Multiple intratumoral staining patterns was observed in primary tumor, and CD44v6 expression in invasive front was significantly higher than that in tumor center. In addition, mRNA expression levels differed across different geographical regions of the tumor. No association between CD44v6 expression and lymph node metastasis was revealed. CONCLUSIONS: Substantial intratumoral heterogeneity of CD44v6 exists in rectal cancer that impacts the outcome of individual studies. CD44v6 expression should be assessed in a more precise way with a specified staining pattern and in a designated location.

Association between transcription factor 7-like 2 genetic polymorphisms and development of type 2 diabetes in a Chinese population.

We conducted a hospital-based case-control study to evaluate the relationship between the transcription factor 7-like 2 (TCF7L2) rs7903146 polymorphism and type 2 diabetes mellitus risk in a Chinese population. Genotyping of TCF7L2 rs7903146 was carried out using the polymerase chain reaction-restriction fragment length polymorphism method. A chi-square test revealed a statistically significant difference between the distributions of rs7903146 genotypes in type 2 diabetes mellitus patient and control groups (chi-square = 10.49, P = 0.005). Using unconditional logistic regression analysis, we observed that the TT genotype of this polymorphism was significantly correlated with increased risk of developing type 2 diabetes mellitus compared to the CC genotype [odds ratio (OR) = 2.31, 95% confidence interval (CI) = 1.33-4.04]. Furthermore, we found that the rs7903146 sequence variation was also significantly associated with susceptibility to this disease under dominant (OR = 1.58, 95%CI = 1.09-2.28) and recessive models  (OR = 2.11, 95%CI = 1.25-3.62). We conclude that the TCF7L2 rs7903146 genetic polymorphism is independently associated with the risk of developing type 2 diabetes mellitus under co-dominant, dominant, and recessive models.

Effect of weaning age on cortisol release in piglets.

The effect of weaning age on the adrenal cortex, which plays a vital role in the stress response, is currently unknown. Therefore, plasma adrenocorticotropic hormone (ACTH) and cortisol levels, weights and relative weights of adrenal glands, and steroidogenesis-related protein and enzyme expression levels in piglets weaned on different days were determined. Piglets weaned at 35 days had significantly lower ACTH levels than those weaned at 14 or 21 days, and cortisol levels of piglets weaned at 21, 28, and 35 days were significantly lower than those of piglets weaned on day 14. Adrenal gland weights of piglets weaned at 28 and 35 days and relative adrenal gland weights of piglets weaned at 35 days were significantly lower than those of piglets weaned at 14 days. However, no significant difference was detected in the expression of melanocortin-type 2 receptor mRNA, which is associated with weaning age. Steroidogenic acute-regulatory (StAR) mRNA and cholesterol side-chain cleavage cytochrome P450 mRNA expression levels in piglets weaned at 28 and 35 days were significantly lower than in those weaned at 14 or 21 days, and P450 11ß mRNA expression levels in piglets weaned at 28 and 35 days were significantly lower than in those weaned at 14 days. Therefore, early-weaned piglets exhibited increased adrenal gland weights and StAR and steroidogenic enzyme expression, all of which contributed to high cortisol levels. The high plasma ACTH and cortisol levels in early-weaned piglets indicate that these animals would be greatly affected by stress.

Prevalence of honeybee viruses in different regions of China and Argentina.

Honeybees are threatened by various pathogens and parasites. More than 18 viruses have been described in honeybees and many of them have been detected in China and Argentina. In China, both Apis cerana and Apis mellifera are raised. In Argentina, beekeepers raise different ecotypes of A. mellifera: European honeybees (in both temperate and subtropical regions) and Africanised honeybees (in subtropical areas only). A thorough study was carried out in both China and Argentina to analyse the current virus presence and distribution in different climatic zones and gather information on different bee species/subspecies. Adult honeybees were collected from apiaries in temperate and subtropical regions of China (including areas with exclusive populations of A. mellifera, areas where A. mellifera and A. cerana co-exist, and areas with exclusive populations of A. cerana) and Argentina. Six viruses, namely, deformed wing virus (DWV), black queen cell virus (BQCV), sacbrood virus (SBV), chronic bee paralysis virus (CBPV), acute bee paralysis virus (ABPV) and Israeli acute paralysis virus (IAPV) were detected in China, both in A. cerana and in A. mellifera, while four viruses (DWV, BQCV, CBPV and ABPV) were present in Argentina. Interestingly, multiple infections were commonly found in China, with up to five different viruses co-circulating in some colonies without apparent abnormalities. In this study, no Chinese samples were positive for slow bee paralysis virus. The most prevalent viruses were BQCV (China) and DWV (Argentina). Kashmir bee virus was absent from samples analysed for both countries.

Les populations d'abeilles mellifères sont menacées par de nombreux agents pathogènes et parasites. Parmi eux, 18 virus ont été décrits, dont plusieurs ont été détectés en Chine et en Argentine. Les espèces d'abeilles mellifères élevées en Chine sont Apis cerana et Apis mellifera. En Argentine, les apiculteurs élèvent plusieurs écotypes d'A. mellifera : le type européen dans les régions tempérées et subtropicales et le type africanisé dans les zones subtropicales. Une étude approfondie a été réalisée en Chine et en Argentine dans le but d'identifier les virus présents, d'analyser leur distribution dans différentes zones climatiques et de réunir des informations sur les différentes espèces et sous-espèces d'abeilles présentes. Des abeilles mellifères adultes ont été collectées dans des ruchers des régions tempérées et subtropicales de Chine (zones peuplées exclusivement d'A. mellifera ou d'A. cerana et zones où A. mellifera et A. cerana coexistent) et d'Argentine (A. mellifera seulement). En Chine, six virus, à savoir le virus des ailes déformées, le virus des cellules royales noires, le virus du couvain sacciforme, le virus de la paralysie chronique de l'abeille, le virus de la paralysie aiguë de l'abeille et le virus israélien de la paralysie aiguë ont été détectés aussi bien chez A. cerana que chez A. mellifera ; en Argentine, quatre virus ont été détectés (virus des ailes déformées, virus des cellules royales noires, virus de la paralysie chronique de l'abeille et virus de la paralysie aiguë de l'abeille). Fait intéressant, les infections multiples étaient fréquentes en Chine, avec parfois jusqu'à cinq virus différents circulant dans certaines colonies sans provoquer de manifestations anormales apparentes. Aucun des échantillons analysés en Chine n'a été trouvé positif pour le virus de la paralysie lente de l'abeille. Les virus les plus fréquents étaient, en Chine, le virus des cellules royales noires et en Argentine, le virus des ailes déformées. Le virus du Cachemire n'a été trouvé dans aucun des échantillons analysés dans les deux pays.

Las abejas melíferas están amenazadas por diversos patógenos y parásitos. Se han descrito más de 18 virus que las afectan, muchos de los cuales se han detectado en China y la Argentina. En China se cultivan tanto Apis cerana como Apis mellifera, mientras que los apicultores argentinos crían diferentes ecotipos de A. mellifera: abejas europeas en las regiones templadas y subtropicales y abejas africanizadas en las zonas subtropicales. Los autores exponen un minucioso estudio realizado a la vez en China y la Argentina con el fin de analizar la actual presencia y distribución de virus en diferentes zonas climáticas y reunir información sobre distintas especies y subespecies de abeja. En primer lugar se recogieron abejas adultas de colmenares situados en regiones templadas y subtropicales de China (algunas donde hay exclusivamente poblaciones de A. mellifera, otras donde coexisten A. mellifera y A. cerana y otras zonas que albergan solo poblaciones de A. cerana) y la Argentina (solamente A. mellifera). En las poblaciones chinas tanto de A. cerana como de A. mellifera se detectaron seis virus: virus de las alas deformes (VAD); virus de las celdas reales negras (VCRN); virus de la cría ensacada (VCE); virus de la parálisis crónica de la abeja (VPCA); virus de la parálisis aguda de la abeja (VPAA); y virus de la variante israelí del virus de la parálisis aguda (VPAI), mientras que en la Argentina se observó la presencia de cuatro virus (VAD, VCRN, VPCA y VPAA). Un dato interesante es que en China se observaron con frecuencia infecciones múltiples, con hasta cinco virus diferentes circulando a la vez en algunas colonias sin que ello diera lugar a anormalidades aparentes. Ninguna de las muestras chinas analizadas en el estudio resultó positiva al virus de la parálisis lenta de la abeja. Los virus más prevalentes fueron el VCRN (China) y el VAD (Argentina). El virus Cachemira de las abejas estaba ausente de las muestras analizadas en ambos países.

Genetic diversity and genetic structure of consecutive breeding generations of golden mandarin fish (Siniperca scherzeri Steindachner) using microsatellite markers.

In this study, 12 polymorphic microsatellites were inves-tigated to determine the genetic diversity and structure of 5 consecu-tive selected populations of golden mandarin fish (Siniperca scherzeri Steindachner). The total numbers of alleles, average heterozyosity, and average polymorphism information content showed that the genetic diversity of these breeding populations was decreasing. Additionally, pairwise fixation index FST values among populations and Da values in-creased from F1 generation to subsequent generations (FST values from 0.0221-0.1408; Da values from 0.0608-0.1951). Analysis of molecular variance indicated that most genetic variations arise from individuals within populations (about 92.05%), while variation among populations accounted for only 7.95%. The allele frequency of the loci SC75-220 and SC101-222 bp changed regularly in the 5 breeding generations. Their frequencies were gradually increased and showed an enrichment trend, indicating that there may be genetic correlations between these 2 loci and breeding traits. Our study indicated that microsatellite markers are effective for assessing the genetic variability in the golden mandarin fish breeding program.

Investigation of ERCC1 and ERCC2 gene polymorphisms and response to chemotherapy and overall survival in osteosarcoma.

We assessed the role of single nucleotide polymorphisms (SNPs) in ERCC1 and ERCC2 genes in the clinical outcomes for osteosarcoma patients receiving cisplatin-based treatment. A perspective study was conducted on 260 patients with osteosarcoma during 2010 and 2011. A polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) assay was used to assess the ERCC1 rs11615 and rs3212986, and the ERCC2 rs1799793 and rs13181 gene polymorphisms. After adjustment for clinical variables, we found that the CC genotype of ERCC1 rs11615 was significantly associated with better response to chemotherapy (OR = 2.87, 95%CI = 1.24-6.97). Our study found that those carrying the CC genotype of ERCC1 rs11615 had a longer overall survival compared with the TT genotype, and the OR (95%CI) was 0.35 (0.12-0.92). In conclusion, our results suggest that the ERCC1 rs11615 polymorphism might influence the response to cisplatin-based chemotherapy and affect the clinical outcome for osteosarcoma patients.

Microsatellite analysis of genetic diversity and genetic structure in five consecutive breeding generations of mandarin fish Siniperca chuatsi (Basilewsky).

In this report, 10 polymorphic microsatellites were applied to assess the genetic diversity and genetic differentiation of 5 consecutive breeding generations of mandarin fish, Siniperca chuatsi (Basilewsky). The results from total number of alleles, average polymorphism information content, and average homozygosity and heterozygosity showed that the genetic diversity of the breeding population was decreasing. The genetic identity between F1 and its descendant generations (F2, F3, F4, F5) decreased (from 0.9248 to 0.8803), while the genetic distance (from 0.0782 to 0.1275) and fixation index (from 0.03796 to 0.07393) increased. The allele frequency of SS181-235 and SS211-246 changed regularly in the 5 breeding generations, and they may be negatively associated with the selected trait, which needs to be confirmed by further research. Our study indicated that selective breeding was an efficient strategy for mandarin fish. In the process of breeding, some deleterious genes were phased out, and the genetic structure of the breeding populations became stable.

Correlation analysis of the PNPLA7 gene polymorphism and susceptibility to menstrual disorder.

We examined the correlation between PNPLA7 gene polymorphisms at the rs61754920 and rs11137410 loci and menstrual disorder in women of reproductive age in the Central Plain. Genomic DNA was extracted from peripheral blood; polymerase chain reaction-ligase detection reaction and SNaPshot genotyping were used to detect polymorphisms in the rs61754920 and rs11137410 gene loci, respectively. The results for the 2 loci in individuals of different blood types were statistically analyzed. The proportion of the AA homozygote at the rs61754920 locus in the PNPLA7 gene was the lowest, while the proportion of the CC homozygote at the rs11137410 locus in the PNPLA7 gene was the highest. There were no statistical differences in the frequency distribution of genotypes and alleles at the 2 loci between control and test groups. The frequency of the TT genotype at the rs11137410 locus in women with type O blood was significantly lower in the test group than in the control group. Frequencies of the C and T alleles were significantly different between the 2 groups. There may be an association between the PNPLA7 gene and type O blood or a combined effect of the 2 genes.

Relationship between the cholesterol ester transfer protein TaqIB polymorphism and the lipid-lowering effect of atorvastatin in patients with coronary atherosclerotic heart disease.

This study aimed to investigate the relationship between the cholesterol ester transfer protein (CETP) gene TaqIB polymorphism and the lipid-lowering effect of atorvastatin in patients with coronary atherosclerotic heart disease. Two hundred eighty-eight patients were divided into a control group, an acute coronary syndrome (ACS) group, and a stable coronary heart disease (CHD) group. Blood biochemical indices were determined using the enzyme method, and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis was performed to study the TaqIB polymorphism of the CETP gene. The ACS and stable CHD groups were treated with atorvastatin, and blood lipid levels were reexamined after three months. Plasma levels of total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), and lipoprotein(a) were all significantly higher in the ACS and stable CHD groups compared to the control group (P < 0.05 or P < 0.01). After three months of treatment with atorvastatin, plasma levels of TC, LDL-C, triglycerides (TG) (only in patients with genotype B2B2), and lipoprotein(a) (only in patients with genotype B1B2) were all significantly decreased (P < 0.05 or P < 0.01). After treatment, the plasma level of TG was lower in patients with genotype B2B2 compared to patients with genotypes B1B1 or B1B2 (B1 carriers) (P < 0.01). Therefore, the CETP TaqIB polymorphism is associated with the lipid-lowering effect of atorvastatin in patients with CHD.

The ratio of transforming growth factor-ß1/bone morphogenetic protein-7 in the progression of the epithelial-mesenchymal transition contributes to rat liver fibrosis.

This study was designed to show whether rat liver epithelial cells could undergo epithelial-mesenchymal transition (EMT), thereby directly contributing to liver fibrosis. The role of the ratio of transforming growth factor-ß1 (TGF-ß1)/bone morphogenetic protein-7 (BMP-7) was evaluated in the progression of EMT or mesenchymal-epithelial transition. Primary rat liver epithelial cells were stimulated with different ratios of TGF-ß1/BMP-7 and examined for evidence of transition to a mesenchymal or epithelial phenotype. Liver sections were labeled to detect antigens associated with liver epithelial cells [E-cadherin (E-cad)], EMT [fibroblast-specific protein-1 (FSP-1), vimentin], myofibroblasts [α-smooth muscle actin (α-SMA)], and intracellular signal-transduction mediated by forming liver fibrosis undergo EMT, resulting in the formation of invasive fibroblasts; this process may be driven or impeded by a response to local TGF-ß1 or BMP-7. BMP-7 downregulated α-SMA and phosphorylated Smad2/3. Stimulation of cultured cells with TGF-ß1 induced the expression of pSmad2/3, FSP-1, and α-SMA. Stimulation of cultured cells with BMP-7 induced the expression of E-cad. We demonstrated that the cells upregulated E-cad release compared with untreated cells, but TGF-ß1 was different. We found that the equilibrium of the ratio of TGF-ß1/BMP-7 was 1/10. In summary, the mechanism for this process was not determined. Demonstration of the contribution of what the ratio of TGF-ß1/BMP-7 induced to EMT to the chronic liver diseases would provide a new basis for understanding pathogenesis and potential treatment.

Expression profile of insulin-like growth factor system genes in muscle tissues during the postnatal development growth stage in ducks.

Insulin-like growth factors (IGFs) are regulators that modulate the proliferation and differentiation of muscle tissues. We quantified the messenger RNA (mRNA) expression of IGF-I, IGF-II, and type I and II IGF receptors (IGF-IR and IGF-IIR) in muscle tissues including the breast, leg, and myocardium during an early postnatal development growth stage (post-hatching weeks 1-8) in ducks. The results showed a significant age-related change in mRNA in these muscle tissues. In breast muscle, the developmental expression of IGF-I and IGF-II was highest during week 1 but decreased quickly and maintained a relatively lower level. Leg muscle had the highest mRNA expression of IGF-I and IGF-II genes at week 3. In myocardial tissues, the expression level of IGF-IR and IGF-IIR genes exhibited a "rise-decline" developmental trend. The expression patterns of IGF-I/IGF-IR and IGF-II/IGF-IIR were different between weeks 4 and 6. The same expression pattern was observed for IGF-I and IGF-IR; however, it was different from that observed for IGF-II and IGF-IIR. Our results showed a negative correlation between IGF-II mRNA expression and leg muscle weight at week 4 (P < 0.05). A negative correlation was also found between IGF-II mRNA expression and breast muscle weight (P < 0.01), and a positive correlation was found between IGF-IR expression and breast muscle weight. At week 6, a positive correlation was found between IGF-IR expression and breast muscle weight. However, at week 8, a negative correlation was found between IGF-IR expression and breast muscle weight. The results showed that the expression of IGF mRNA in duck tissues exhibits a specific developmental trend and an age-related pattern, suggesting that the regulation mechanism of these 4 genes in proliferation and differentiation of muscle tissues differed.