H2S exposure-induced oxidative stress promotes LPS-mediated hepatocyte autophagy through the PI3K/AKT/TOR pathway.

Hydrogen sulfide (H2S), a common air pollutant and toxic gas, is detrimental to organisms and the environment. Exposure to highly concentrated H2S can induce oxidative stress and autophagy. However, the mechanism underlying the liver damage caused by H2S has not been identified. Lipopolysaccharide (LPS), the key component of endotoxin, can induce oxidative stress and autophagy. For this experiment, we used one-day-old chickens as model organisms to evaluate the effects of H2S combined with LPS on oxidative stress and autophagy. The four groups (control group, LPS group, H2S group and H2S-LPS group) were observed by electron microscopy, detected by oxidative stress kit, analyzed by quantitative real-time quantitative PCR, and analyzed by Western blot. We found that the activities of antioxidant enzymes (superoxide dismutase, antioxidant glutathione, catalase, and glutathione peroxidase) decreased in the H2S group compared to those in the control group; however, malondialdehyde levels in the H2S group increased. Molecular-level studies showed that the expression of genes associated with the PI3K/ AKT/ TOR pathways in the H2S group decreased, whereas the expression of other autophagy-related genes (Beclin1, ATG5 and the ratio of LC3-II/ LC3-I) increased compared to that in the control group. These findings suggest that H2S caused oxidative stress and induced autophagy through the PI3K/ AKT/ TOR pathway in chicken liver cells. Additionally, exposure to H2S aggravated LPS-induced oxidative stress and autophagy injury. Capsule: Aerial exposure to H2S can cause oxidative stress in chicken livers and induce autophagy through the PI3K/AKT/TOR pathway, and can aggravate LPS-induced oxidative stress and autophagy.

Application of 3-Dimensional Computerized Tomography Angiography for Defining Cavernous Sinus Aneurysms and Intradural Aneurysms Involving the Internal Carotid Artery Around the Anterior Clinoid Process.

OBJECTIVE: This study aimed to investigate the application of 3-dimensional computed tomography angiography (3D-CTA) for defining cavernous sinus aneurysms and intradural aneurysms involving the internal carotid artery around the anterior clinoid process. METHODS: Results from 42 patients with an aneurysm of the internal carotid artery around the anterior clinoid process who underwent 3D-CTA were reviewed and compared with those of observed clinical operations. RESULTS: Among the 42 patients, there was a total of 45 aneurysms of the internal carotid artery around the anterior clinoid process. After surgery, 33 of the 45 aneurysms were confirmed as intradural aneurysms, and the other 12 were confirmed as aneurysms in the cavernous sinus. 3D-CTA imaging of the medial sagittal plane showed that 31 out of 31 (100%) intradural aneurysms of the internal carotid artery were above the virtual line between the inferior border of the anterior clinoid process and the tuberculum sellae, and 12 out of 14 (86%) cavernous sinus aneurysms were below the virtual line (P < 0.0001). CONCLUSIONS: The virtual line between the inferior border of the anterior clinoid process and the tuberculum sellae on 3D-CTA indicates the proximal dural ring of the internal carotid artery. This line helps differentiate cavernous sinus aneurysms from intradural aneurysms involving the internal carotid artery around the anterior clinoid process.

Effects of relative humidity on animal health and welfare.

Farm animals are sources of meat, milk and eggs for the humans, and animal health ensures the quality and security of these agricultural and sideline products. The animal raising conditions in livestock stations and poultry houses play vital roles in both animal health and production. One of the major factors affecting raising conditions, relative humidity, has not received much attention even though it is important for animal husbandry. In this review, we summarize the impacts of relative humidity on animal health and welfare to draw attention for its importance in the improvement of animal raising conditions in the future.

Tissue-Specific Regulation of the Contents and Correlations of Mineral Elements in Hens by Zinc Oxide Nanoparticles.

Due to their small size, zinc oxide (ZnO) nanoparticles (NPs) are readily absorbed and easily cross biological barriers, which make them promising candidates as diet additives. However, some studies have reported that ZnO NPs cause toxicity; therefore, their safety and potency as diet additives for farm animals should be established. This study was the first to fully evaluate the effects of ZnO NPs on the homeostasis of eight elements in seven organs/tissues. The regulation of element homeostasis was found to be organ specific with no influence on oxidation status, anti-oxidation capability, or organ damage. ZnO NPs may specifically regulate the homeostasis of mineral elements and affect the following correlations: (1) between the element content in each organ and the concentration of Zn used in ZnSO4 or ZnO NP treatments; (2) between ZnO NP and ZnSO4 treatments for the same element in each organ; and (3) between elements (in each organ in ZnSO4 or ZnO NP treatments) in layers' organs/tissues. The use of ZnO NPs as diet additives for animals should be implemented cautiously because, among other uncertainties, they may affect mineral element content.

Hydrogen Sulfide and/or Ammonia Reduces Spermatozoa Motility through AMPK/AKT Related Pathways.

A number of emerging studies suggest that air pollutants such as hydrogen sulfide (H2S) and ammonia (NH3) may cause a decline in spermatozoa motility. The impact and underlying mechanisms are currently unknown. Boar spermatozoa (in vitro) and peripubertal male mice (in vivo) were exposed to H2S and/or NH3 to evaluate the impact on spermatozoa motility. Na2S and/or NH4Cl reduced the motility of boar spermatozoa in vitro. Na2S and/or NH4Cl disrupted multiple signaling pathways including decreasing Na+/K+ ATPase activity and protein kinase B (AKT) levels, activating Adenosine 5'-monophosphate (AMP)-activated protein kinase (AMPK) and phosphatase and tensin homolog deleted on chromosome ten (PTEN), and increasing reactive oxygen species (ROS) to diminish boar spermatozoa motility. The increase in ROS might have activated PTEN, which in turn diminished AKT activation. The ATP deficiency (indicated by reduction in Na+/K+ ATPase activity), transforming growth factor (TGFß) activated kinase-1 (TAK1) activation, and AKT deactivation stimulated AMPK, which caused a decline in boar spermatozoa motility. Simultaneously, the deactivation of AKT might play some role in the reduction of boar spermatozoa motility. Furthermore, Na2S and/or NH4Cl declined the motility of mouse spermatozoa without affecting mouse body weight gain in vivo. Findings of the present study suggest that H2S and/or NH3 are adversely associated with spermatozoa motility.

Determinants of undergraduate nursing students' care willingness towards the elderly in China: Attitudes, gratitude and knowledge.

BACKGROUND: An aging population has become a serious problem in China. Improving the nursing students' care willingness is a critical way to solve this dilemma. Few studies reveal the relationship between the knowledge, care willingness, attitude towards the elderly and gratitude. This research has found that the attitude towards the elderly, the knowledge about aging, and gratitude showed correlation with care willingness. OBJECTIVE: The purpose of this study is to explore the relationships among knowledge about aging, care willingness, attitude towards the elderly and gratitude. DESIGN: A cross-sectional descriptive design has been used. METHODS: From November to December 2015, a total of 382 undergraduate nursing students in China completed the questionnaires. The response rate was 95.5%. Four questionnaires including Care Willingness to the Elderly Scale (CW), Kogan's Attitudes towards Old People scale (KAOP), the Facts on Aging Quiz (FAQ), and the Gratitude Scale. Structural equation modeling (SEM) was used to explore the relationship among those variables in this study. RESULTS: For Chinese nursing students, the care willingness of elderly was in medium-high level. Their attitude towards the elderly and gratitude were at the medium degree, while the knowledge about aging was at a lower level. The attitude towards older people, knowledge about aging, and gratitude were significantly correlated with care willingness. The knowledge about aging has no relationship with the attitude. Gratitude plays a mediation role between the knowledge about aging and care willingness. The experience of caring the elderly could lead to a positive impact in care willingness. CONCLUSIONS: The nursing students' knowledge about aging had a direct influence on their care willingness. Gratitude plays a mediating role between the knowledge about aging and care willingness to the elderly.

Regulation of MicroRNAs, and the Correlations of MicroRNAs and Their Targeted Genes by Zinc Oxide Nanoparticles in Ovarian Granulosa Cells.

Zinc oxide (ZnO) nanoparticles (NPs) have been applied in numerous industrial products and personal care products like sunscreens and cosmetics. The released ZnO NPs from consumer and household products into the environment might pose potential health issues for animals and humans. In this study the expression of microRNAs and the correlations of microRNAs and their targeted genes in ZnO NPs treated chicken ovarian granulosa cells were investigated. ZnSO4 was used as the sole Zn2+ provider to differentiate the effects of NPs from Zn2+. It was found that ZnO-NP-5 µg/ml specifically regulated the expression of microRNAs involved in embryonic development although ZnO-NP-5 µg/ml and ZnSO4-10 µg/ml treatments produced the same intracellular Zn concentrations and resulted in similar cell growth inhibition. And ZnO-NP-5 µg/ml also specifically regulated the correlations of microRNAs and their targeted genes. This is the first investigation that intact NPs in ZnO-NP-5 µg/ml treatment specifically regulated the expression of microRNAs, and the correlations of microRNAs and their targeted genes compared to that by Zn2+. This expands our knowledge for biological effects of ZnO NPs and at the same time it raises the health concerns that ZnO NPs might adversely affect our biological systems, even the reproductive systems through regulation of specific signaling pathways.

Regulation of neuroendocrine cells and neuron factors in the ovary by zinc oxide nanoparticles.

The pubertal period is an important window during the development of the female reproductive system. Development of the pubertal ovary, which supplies the oocytes intended for fertilization, requires growth factors, hormones, and neuronal factors. It has been reported that zinc oxide nanoparticles (ZnO NPs) cause cytotoxicity of neuron cells. However, there have been no reports of the effects of ZnO NPs on neuronal factors and neuroendocrine cells in the ovary (in vivo). For the first time, this in vivo study investigated the effects of ZnO NPs on gene and protein expression of neuronal factors and the population of neuroendocrine cells in ovaries. Intact NPs were detected in ovarian tissue and although ZnO NPs did not alter body weight, they reduced the ovary organ index. Compared to the control or ZnSO4 treatments, ZnO NPs treatments differentially regulated neuronal factor protein and gene expression, and the population of neuroendocrine cells. ZnO NPs changed the contents of essential elements in the ovary; however, they did not alter levels of the steroid hormones estrogen and progesterone. These data together suggest that intact ZnO NPs might pose a toxic effect on neuron development in the ovary and eventually negatively affect ovarian developmental at puberty.

Alteration of gene expression by zinc oxide nanoparticles or zinc sulfate in vivo and comparison with in vitro data: A harmonious case.

Granulosa cells (GCs) are those somatic cells closest to the female germ cell. GCs play a vital role in oocyte growth and development, and the oocyte is necessary for multiplication of a species. Zinc oxide (ZnO) nanoparticles (NPs) readily cross biologic barriers to be absorbed into biologic systems that make them promising candidates as food additives. The objective of the present investigation was to explore the impact of intact NPs on gene expression and the functional classification of altered genes in hen GCs in vivo, to compare the data from in vivo and in vitro studies, and finally to point out the adverse effects of ZnO NPs on the reproductive system. After a 24-week treatment, hen GCs were isolated and gene expression was quantified. Intact NPs were found in the ovary and other organs. Zn levels were similar in ZnO-NP-100 mg/kg- and ZnSO4-100 mg/kg-treated hen ovaries. ZnO-NP-100 mg/kg and ZnSO4-100 mg/kg regulated the expression of the same sets of genes, and they also altered the expression of different sets of genes individually. The number of genes altered by the ZnO-NP-100 mg/kg and ZnSO4-100 mg/kg treatments was different. Gene Ontology (GO) functional analysis reported that different results for the two treatments and, in Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment, 12 pathways (out of the top 20 pathways) in each treatment were different. These results suggested that intact NPs and Zn(2+) had different effects on gene expression in GCs in vivo. In our recent publication, we noted that intact NPs and Zn(2+) differentially altered gene expression in GCs in vitro. However, GO functional classification and KEGG pathway enrichment analyses revealed close similarities for the changed genes in vivo and in vitro after ZnO NP treatment. Furthermore, close similarities were observed for the changed genes after ZnSO4 treatments in vivo and in vitro by GO functional classification and KEGG pathway enrichment analyses. Therefore, the effects of ZnO NPs on gene expression in vitro might represent their effects on gene expression in vivo. The results from this study and our earlier studies support previous findings indicating ZnO NPs promote adverse effects on organisms. Therefore, precautions should be taken when ZnO NPs are used as diet additives for hens because they might cause reproductive issues.

iTRAQ-based quantitative proteomic analysis of longissimus muscle from growing pigs with dietary supplementation of non-starch polysaccharide enzymes.

Non-starch polysaccharide enzymes (NSPEs) have long been used in the feed production of monogastric animals to degrade non-starch polysaccharide to oligosaccharides and promote growth performance. However, few studies have been conducted on the effect of such enzymes on skeletal muscle in monogastric animals. To elucidate the mechanism of the effect of NSPEs on skeletal muscle, an isobaric tag for relative and absolute quantification (iTRAQ) for differential proteomic quantitation was applied to investigate alterations in the proteome in the longissimus muscle (LM) of growing pigs after a 50-d period of supplementation with 0.6% NSPEs in the diet. A total of 51 proteins were found to be differentially expressed in the LM between a control group and the NSPE group. Functional analysis of the differentially expressed protein species showed an increased abundance of proteins related to energy production, protein synthesis, muscular differentiation, immunity, oxidation resistance and detoxification, and a decreased abundance of proteins related to inflammation in the LM of the pigs fed NSPEs. These findings have important implications for understanding the mechanisms whereby dietary supplementation with NSPEs enzymes can promote growth performance and improve muscular metabolism in growing pigs.

Screening and monitoring zearalenone-producing Fusarium species by PCR and zearalenone by monoclonal antibodies in feed from China.

Screening of zearalenone (ZEN)-producing species and monitoring of ZEN in feed were performed by using anti-zearalenone monoclonal antibodies. ELISA recoveries of ZEN from corn distillers dried grains with solubles (DDGS) feed, corn feed, rice bran, soybean meal, wheat bran and rapeseed dregs were between 78.6% and 88.6%. ZEN recovery from culture media was 96.3% at the spiked level of 500 µg/kg. Eighty-three samples of DDGS feed, corn feed and other fee ingredients were collected from 11 provinces of China and analysed for ZEN. Average ZEN levels were 563.4 µg/kg for DDGS feed, 393.1 µg/kg for corn feed and 65.7 µg/kg for other feed ingredients. Eighteen Fusarium species such as Fusarium graminearum which could produce ZEN were isolated from corn feed and other feed ingredients.

Abnormal lithium isotope composition from the ancient lithospheric mantle beneath the North China Craton.

Lithium elemental and isotopic compositions of olivines in peridotite xenoliths from Hebi in the North China Craton provide direct evidence for the highly variable δ(7)Li in Archean lithospheric mantle. The δ(7)Li in the cores of olivines from the Hebi high-Mg# peridotites (Fo > 91) show extreme variation from -27 to +21, in marked deviation from the δ(7)Li range of fresh MORB (+1.6 to +5.6) although the Li abundances of the olivines are within the range of normal mantle (1-2 ppm). The Li abundances and δ(7)Li characteristics of the Hebi olivines could not have been produced by recent diffusive-driven isotopic fractionation of Li and therefore the δ(7)Li in the cores of these olivines record the isotopic signature of the subcontinental lithospheric mantle. Our data demonstrate that abnormal δ(7)Li may be preserved in the ancient lithospheric mantle as observed in our study from the central North China Craton, which suggest that the subcontinental lithospheric mantle has experienced modification of fluid/melt derived from recycled oceanic crust.

Risks associated with high-dose Lactobacillus rhamnosus in an Escherichia coli model of piglet diarrhoea: intestinal microbiota and immune imbalances.

Probiotic could be a promising alternative to antibiotics for the prevention of enteric infections; however, further information on the dose effects is required. In this study, weanling piglets were orally administered low- or high-dose Lactobacillus rhamnosus ACTT 7469 (10(10) CFU/d or 10(12) CFU/d) for 1 week before F4 (K88)-positive Escherichia coli challenge. The compositions of faecal and gastrointestinal microbiota were recorded; gene expression in the intestines was assessed by real-time PCR; serum tumour necrosis factor-α (TNF-α) concentrations and intestinal Toll-like receptor 4 (TLR4) were detected by ELISA and immunohistochemistry, respectively. Unexpectedly, high-dose administration increased the incidence of diarrhoea before F4(+)ETEC challenge, despite the fact that both doses ameliorated F4(+)ETEC-induced diarrhoea with increased Lactobacillus and Bifidobacterium counts accompanied by reduced coliform shedding in faeces. Interestingly, L. rhamnosus administration reduced Lactobacillus and Bifidobacterium counts in the colonic contents, and the high-dose piglets also had lower Lactobacillius and Bacteroides counts in the ileal contents. An increase in the concentration of serum TNF-α induced by F4(+)ETEC was observed, but the increase was delayed by L. rhamnosus. In piglets exposed to F4(+)ETEC, jejunal TLR4 expression increased at the mRNA and protein levels, while jejunal interleukin (IL)-8 and ileal porcine ß-defensins 2 (pBD2) mRNA expression increased; however, these increases were attenuated by administration of L. rhamnosus. Notably, expression of jejunal TLR2, ileal TLR9, Nod-like receptor NOD1 and TNF-α mRNA was upregulated in the low-dose piglets after F4(+)ETEC challenge, but not in the high-dose piglets. These findings indicate that pretreatment with a low dose of L. rhamnosus might be more effective than a high dose at ameliorating diarrhoea. There is a risk that high-dose L. rhamnosus pretreatment may negate the preventative effects, thus decreasing the prophylactic benefits against potential enteric pathogens. Our data suggest a safe threshold for preventative use of probiotics in clinical practice.

[Expressions of 6 microRNAs in prostate cancer].

OBJECTIVE: Experimental evidence shows that microRNAs play an important role in the initiation and progression of human malignancies. The present study aimed to investigate the expressions of 6 microRNAs in prostate cancer (PCa) and their clinical significance. METHODS: We investigated the expression profiles of 6 microRNAs (let-7g, let-7d, miR-98, miR-96, miR-182 and miR-183) using the method of locked nucleic acid (LNA)-modified oligonucleotide in situ hybridization (ISH) and the technology of tissue microarray (TMA) with the formalin-fixed paraffin-embedded (FFPE) specimens from 52 patients with PCa and 38 with benign prostatic hyperplasia (BPH). Then we analyzed the correlation among the expressions of the 6 microRNAs in PCa and their correlation with the Gleason score and clinical stages of PCa. RESULTS: Compared with BPH, the PCa patients showed decreased expressions of miR-98, let-7d and let-7g, and decreased expressions of miR-96, miR-182 and miR-183, with statistically significant differences between the two groups (P < 0.05). The positive rate of the 6 microRNAs was significantly correlated with the Gleason grades of PCa (P < 0.05), but not with the age and serum PSA concentration of the patients (P > 0.05). The expressions of miR-96 and miR-182 were correlated with the clinical stages of the tumor (P < 0.05). There was a positive correlation among the expressions of miR-96, miR-182 and miR-183 (P = 0.00, r = 0.41), as well as between the expressions of let-7d and let-7g (P = 0.00, r = 0.46) in the PCa tissues. And the expression of miR-98 was positively correlated with those of let-7d and let-7g (P = 0.00, r = 0.46). CONCLUSION: The expression profiles of the microRNAs let-7d, let-7g, miR-98, miR-96, miR-182 and miR-183 reflect the biological behavior of PCa to some extent, and might be important biomarkers for the early detection and prognostic assessment of prostate cancer.

Blocking effects of siRNA on VEGF expression in human colorectal cancer cells.

AIM: To investigate the expression of vascular endothelial cell growth factor (VEGF) and its receptors Fms-like tyrosine kinase 1 (FLT-1) and fetal liver kinase 1 (FLK-1) in colorectal carcinoma (CRC), and the blocking effects of small interfering RNAs (siRNAs) on VEGF expression in human colorectal cancer HCT116 cells. METHODS: Immunohistochemical staining for VEGF, FLT-1 and FLK-1 proteins was performed in 82 cases of CRC and 14 normal colorectal mucosae. A siRNA targeting VEGF was synthesized and transfected into HCT116 cells using lipofectamine 2000. Immunocytochemical staining and Western blotting analyses were performed to detect the expression of VEGF protein. The suppressive effect of the siRNA on cell proliferation was detected using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltertrazolium bromide (MTT) assay. Cellular apoptosis was detected using flow cytometry (FCM). RESULTS: The expression of VEGF, FLT-1 and FLK-1 in tumor tissues was significantly higher than that in normal tissues (P = 0.008, P = 0.000, P = 0.000). The expression of VEGF was positively correlated with both lymph node metastasis and clinical stage (P = 0.009 and P = 0.025, respectively). Immunocytochemistry showed that the expression of VEGF was weakly positive and Western blotting indicated a significant reduction in VEGF-siRNA cell protein levels. VEGF-siRNA cell growth inhibition was assessed by the MTT assay, and the tumor cell proliferation rate was significantly different at 24, 48, and 72 h after transfection. FCM results showed that the VEGF-siRNA group had an apparent aneuploid peak. CONCLUSION: VEGF, FLT-1 and FLK-1 are associated with colorectal carcinogenesis. siRNA silencing of the VEGF gene suppresses proliferation, and induces apoptosis in HCT116 cells. The results suggest that VEGF may be a new gene therapy target for colorectal cancer.

Expression and clinical significance of S100A2 and p63 in esophageal carcinoma.

AIM: To investigate the expression and clinical significance of S100A2 mRNA and protein, p63 protein in esophageal squamous cell carcinoma (ESCC) and their roles in carcinogenesis and progression of esophageal carcinoma (EC). METHODS: Immunohistochemical staining (S-P method) for S100A2 and p63 protein were performed in 40 samples of ESCC and 40 samples of normal esophageal mucosa. In situ hybridization (ISH) was used to detect the expression of S100A2 mRNA. RESULTS: Expression of S100A2 mRNA in ESCC was positive in 77.5% of samples, which was lower than that in normal mucosa (100%) by ISH (P = 0.002). The expression level of S100A2 mRNA was closely related to differentiation and and node-metastasis (P = 0.012, P = 0.008). Expression of S100A2 protein was positive in 72.5% of ESCC samples and expression of p63 protein was positive in 37.5% of ESCC samples, and was lower than that in normal mucosa (100%) (P = 0.000). The expression of S100A2 protein was correlated with the differentiation and node-metastasis (P = 0.007, P = 0.001), but no relationship was observed between the expression of p63 protein and clinical pathological manifestations. S100A2 protein was positively correlated with the expression of S100A2 mRNA, and negatively associated with the expression of p63 protein (P = 0.000, P = 0.002). CONCLUSION: S100A2 and p63 protein both play important roles in the carcinogenesis of ESCC. An investigation into the combined expression of S100A2 and p63 may be helpful in early diagnosis and in evaluating the prognosis of ESCC.

Trichloroethylene induced cutaneous irritation in BALB/c hairless mice: histopathological changes and oxidative damage.

Trichloroethylene (TCE), a colorless and volatile organic solvent, has long been a major chemical hazard during occupational exposure because of its extensive use in industry. Exposure to TCE can cause significant skin lesions, but the effect of TCE on skin irritation has received little attention. We therefore investigated the effect of TCE on skin irritation and oxidative stress using hairless mice. BALB/c hairless mice were subjected to acute and cumulative topical TCE treatment. Skin reactions were evaluated by visual inspection, histopathology examined by microscopy and oxidative stress assessed by measurement of malondialdehyde (MDA) levels, superoxide dismutase (SOD) activities and nitric oxide (NO) production. Under acute and cumulative TCE irritation, the skin developed erythema and edema, and the predominant histopathological features were hyperkeratosis, spongiosis and inflammatory cell infiltrates. In parallel to these morphological changes, acute TCE irritation also concentration-dependently increased MDA levels and inhibited SOD activities of the skin. However, in cumulative irritation, the MDA levels and SOD activities were initially elevated with increased TCE concentrations, but thereafter reduced with further concentration increments; the linear dose-response relationship was not preserved. TCE also concentration-dependently increased NO production both in acute and cumulative irritation. These results suggest that TCE is capable of producing skin irritation effect in vivo, with histopathological changes characterized by hyperkeratosis, spongiosis and inflammatory cell infiltrates. Moreover, oxidative stress may be associated with the clinical manifestations and histopathological abnormalities in TCE-induced skin irritation.

[Expression of human Sel-1-like gene and its significance in esophageal cancer].

OBJECTIVE: To study the expression of SEL1L (human Sel-1-like gene) mRNA and protein and its significance in esophageal cancer. METHODS: Immunohistochemical staining (S-P method) for SEL1L protein was performed in 90 samples of esophageal squamous cell carcinoma, 35 samples of normal esophageal mucosa 5 cm away from the tumor, 60 samples of esophageal mucosa adjacent to the tumor and 20 samples of esophageal squamous dysplasia. In situ hybridization for SEL1L mRNA was also carried out in the esophageal carcinoma cases and normal esophageal mucosa distant from and adjacent to the tumor. RESULTS: The positive rate of SEL1L mRNA was higher in esophageal carcinoma (80.0%, 72/90), as compared with that in normal esophageal mucosa distant from (14.3%, 5/35) and adjacent to (16.7%, 10/60) the tumor (P < 0.01). The positive rate of SEL1L mRNA in tumors with lymph node metastasis (92.7%, 38/41) was higher than that in tumors without lymph node metastasis (69.4%, 34/49) (P < 0.01). On the other hand, the expression rate of SEL1L protein was higher in esophageal carcinoma (87.8%, 79/90) and esophageal dysplasia (90.0%, 18/20), as compared with that in normal esophageal mucosa distant from (14.3%, 5/35) and adjacent to (13.3%, 8/60) the tumor (P < 0.01). The expression of SEL1L protein in esophageal cancer however did not correlate with age and sex of the patient, tumor location, tumor size, degree of differentiation, depth of tumor invasion, lymph node metastasis and tumor clinical stage (P > 0.05). A positive correlation was found between the expression of SEL1L mRNA and SEL1L protein (r = 0.492, P < 0.01). CONCLUSIONS: L1L protein expression is regulated at the transcriptional level. The high SEL1L protein expression is mainly the result of increased transcription. Overexpression of SEL1L protein is likely an early event during the pathogenesis of esophageal squamous cell carcinoma. SEL1L protein may serve as an important biomarker in identifying patients with higher risk of developing esophageal cancer.

Mad2 and p27 expression profiles in colorectal cancer and its clinical significance.

AIM: To investigate the expression of tumor suppressor gene p27 and spindle checkpoint gene Mad2 and to demonstrate their expression difference in colorectal cancer and normal mucosa and to evaluate its clinical significance. METHODS: Immunohistochemical staining was used for detection of expression of Mad2 and p27 in colorectal cancer and its corresponding normal mucosa. RESULTS: Mad2 was significantly overexpressed in colorectal cancer compared with corresponding normal mucosa (P<0.01, chi(2) = 7.5), and it was related to the differentiation of adenocarcinoma, lymph node metastasis and survival period after excision (P<0.05, chi(2) = 7.72, chi(2) = 4.302, chi(2) = 6.234). The rate of p27 positive expression in adenocarcinomas and normal mucosa was 40% and 80% respectively. There was a significant difference in p27 expression between adenocarcinomas and normal mucosa (P<0.001, chi(2) = 13.333), which was related to the differentiation degree of adenoca rcinoma and lymph node metastasis (P<0.05, chi(2) = 8.901 chi(2) = 4). The positive expression of p27 was not correlated with survival period after excision. CONCLUSION: Defect of spindle checkpoint gene Mad2 and mutation of p27 gene are involved mainly in colorectal carcinogenesis and associated with prognosis of colorectal cancer.

Mad2 and p53 expression profiles in colorectal cancer and its clinical significance.

AIM: To investigate the expression of tumor suppressor gene p53 and spindle checkpoint gene Mad2, and to demonstrate their expression difference in colorectal cancer and normal mucosa and to evaluate its clinical significance. METHODS: Western blot and immunohistochemistry methods were used to analyze the expression of Mad2 in colorectal cancer and its corresponding normal mucosa. The expression of p53 was detected by immunohistochemistry method in colorectal cancer and its corresponding normal mucosa. RESULTS: Mad2 was significantly overexpressed in colorectal cancer compared with corresponding normal mucosa (P<0.001), and it was not related to the differentiation of adenocarcinoma and other clinical factors (P>0.05). The ratio of Mad2 protein in cancer tissue (C) to that in its normal mucosa tissue (N) was higher than 2, which was more frequently observed in patients with lymph gland metastasis (P<0.05). p53 protein expression was not observed in normal mucosa. The rate of p53 positive expression in adenocarcinomas was 52.6 %. There was a significant difference between adenocarcinomas and normal mucosa(P<0.001), which was not related to the differentiation degree of adenocarcinoma and other clinical factors (P>0.05). CONCLUSION: Defect of spindle checkpoint gene Mad2 and mutation of p53 gene are involved mainly in colorectal carcinogenesis and C/N>2 is associated with prognosis of colorectal cancer.