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Background: Antibodies to saccharomyces cerevisiae (ASCA), antibodies to perinuclear anti-neutrophil cytoplasmic (pANCA), pancreatic autoantibodies (PAB) and antibodies against intestinal goblet cells (GAB) are important in diagnosing Crohn's disease (CD) and ulcerative colitis (UC). However, little is known about their diagnostic value in real clinical practice in China. This retrospective study aimed to present our 2-year clinical experience with those biomarkers in diagnosis of CD and UC. Methods: A total of 140 patients with UC, 128 patients with CD, and 224 patients with intestinal associated diseases as disease controls were included. Serum ASCA were determined by ELISA. Serum pANCA, GAB, and PAB were tested by indirect immunofluorescent assay. Retrospective review of laboratory results and clinical information was performed. Results: ASCA and ASCA +/pANCA - showed poor abilities in differentiating CD from UC, CD from intestinal Behçet's disease (BD), or CD from intestinal tuberculosis (ITB). In contrast, PAB exhibited good capacities in differentiating CD from UC, CD from intestinal BD, and CD from ITB. IgG pANCA demonstrated a high sensitivity and specificity in differentiating UC from CD. pANCA +/ASCA - or pANCA +/PAB - displayed a high sensitivity and specificity in differentiating UC from CD. GAB showed poor potential in differentiating UC from CD. PAB were positively correlated with early disease onset, ileocolonic disease, and perianal disease in CD patients. Conclusions: Our data suggest that pANCA and PAB are helpful in diagnosis of UC and CD, respectively, while ASCA and GAB were not. Our findings indicate a clear need for additional biomarkers for diagnosis of CD in China. [ABSTRACT FROM AUTHOR]

To understand the mechanisms affecting water balance partitioning during fallow on drylands could improve the fallow management practices in arable land ecosystems. A three-year field experiment was conducted to evaluate the effects of field management regimes on water balance partitioning and fallow efficiency during the fallow periods under a winter wheat (Triticum aestivum L.) fallow system on the Loess Plateau, China. The fallow management regimes tested were: (i) conventional practice, (ii) catch cropping, and (iii) no tillage with wheat straw mulching. A process-oriented ecosystem model (CoupModel) was calibrated with field measurements and then used to generate comparative simulations of the water balance partitioning. The simulations indicated that mulching increased the soil water storage change by 38-71 mm during the three fallow periods, thus resulting in higher fallow efficiency by 9-12%, and decreased soil evaporation by 22-72 mm, compared with the conventional practice. Furthermore, water reached deeper horizons, resulting in 7 mm deep percolation in a wet year under mulching but not under conventional practice or catch cropping. The simulation results also showed that the catch cropping decreased the soil water storage change by 13-21 mm, although it lowered soil evaporation by 11-51 mm, and altogether reduced the fallow efficiency by 3-9%, compared to conventional practice. On the Loess Plateau of China mulching proved to be a sound measure for ensuring certain fallow efficiency and possibly benefit to the water cycle, while catch cropping negatively partitioned the water balance. The catch cropping under mulching might be another management regime to be considered. [ABSTRACT FROM AUTHOR]

To investigate the effects of an Aurora kinase B inhibitor (ZM447439) on the cervical cancer cell line SiHa and chemotherapy of cisplatin (cDDP). Detected Aurora-B protein in different tissues of the cervix by immunohistochemistry and then analyzed the relationship between Aurora B protein and clinical parameters of cervical cancer. The effect and synergistic effect of ZM447439 and cDDP on proliferation of SiHa cells was tested by MTT. The changes of cell cycle and apoptosis were detected by flow cytometry. Aurora-B, histone H3 phosphorylation (H3-P) protein, human papillomavirus16 E6 (HPV16E6) and BCL-2, P53, VEGF protein were detected by Western blot. The positive rate of Aurora-B expression was the highest in cervical cancer and had no significant correlation with clinical stage, lymph node metastasis and age. ZM447439 can reduce the number of SiHa cells, increase the volume of cells and lead to apoptosis. The growth of SiHa cells treated with ZM447439, cDDP and the combination of both was inhibited in dose- and time-dependent manners. The inhibition rate of the combined treatment is significantly higher than that of two other single drug groups ( P < 0.05). The synergistic effect is observed in the combined therapy. S-phase arrest and early apoptosis has become more evident in the combined treatment. ZM447439 significantly inhibited the expression of Aurora-B and H3-P protein ( P < 0.05). ZM447439, cDDP and the combination of both reduced the expression of HPV16E6 and BCL-2 protein, raised P53 protein expression ( P < 0.05), whose effects were more obvious in the combined therapy. cDDP could also reduce VEGF protein expression, but ZM447439 could not. Our results suggest that Aurora-B may represent a valid target in cervical squamous carcinoma and has a synergistic effect with cDDP. [ABSTRACT FROM AUTHOR]

Abstract: Aim of the study: Guizhi-Fuling-decoction (GZFLD), a traditional Chinese medical formulation, exerts an anti-tumor effect, but the mechanisms of its action on invasive tumor inhibition have not been documented. The aims of this study were to identify the inhibitory effect of GZFLD on the invasive of cervical cancer and to elucidate the extensional mechanisms of its action. Materials and method: The invasive ability of HeLa cells was tested with Transwell chamber. The expressions and activities of matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) were measured by zymography/reverse zymography, RT-PCR and Western blot analysis. Establish tumor-bearing mice model to assess the ability of GZFLD to inhibit tumor growth and angiopoiesis in vivo. Results: We have found that GZFLD suppressed the invasive ability of HeLa cells, inhibited MMPs expressions and activities, increased TIMPs expressions and activities, and furthermore restored the MMPs–TIMPs balance in HeLa cells in a concentration-dependent manner. Meanwhile in vivo, GZFLD had significantly inhibited tumor growth and angiopoiesis. Conclusion: In general, our results showed that GZFLD had inhibited the invasion of cervical cancer both in vitro and vivo. The inhibitory effects may be associated with restoring the MMPs–TIMPs balance, and then suppressing the degradation of extracellular matrix. [Copyright &y& Elsevier]

Background: Dickkopf-4 (DKK4), a member of DKK family, appears to be a divergent protein. It remained multi-biological functions in carcinogenesis. The effect of DKK4 on the ovarian cancer cells remains unclear. This study detected the clinical significance of DKK4 in epithelial ovarian cancer (EOC) patients and its role in invasion.Methods: QRT-PCR and western blot analysis were used to examine the levels of DKK4 mRNA and protein in 33 EOC tissues and 33 benign ovarian tumors. Immunohistochemical analysis was performed to assess DKK4 expression in 239 EOC samples. siRNA-mediated DKK4 silence was conducted. Transwell assay was used to detect the invasive ability. Phalloidin was used to stain the formations of actin filaments.Results: The expressions of DKK4 mRNA and protein were elevated in EOC tissues as compared with those in benign ovarian tumors (p = 0.001 and <0.0001 respectively). Immunohistochemical results showed the strong expression of DKK4 protein was positively associated with late FIGO stage (p = 0.005) and poor disease free survival in univariate and multivariate analysis (p < 0.0001 and p = 0.001, respectively). SiRNA-mediated DKK4 knockdown inhibited cell invasive ability (all p < 0.0001) and the formations of actin filaments. DKK4 could promote the phosphration of c-JUN and JNK (p < 0.0001 and p = 0.001, respectively).Conclusions: Our results indicated that DKK4 might be contributed to predicting EOC progression and prognosis. DKK4 could promote the invasion of EOC through JNK activation. [ABSTRACT FROM AUTHOR]

Based on the phase separation phenomenon of acetonitrile-water system at subzero temperature, a subzero-temperature liquid-liquid extraction coupled with ultra-performance liquid chromatography tandem quadrupole mass spectrometry (UPLC-MS-MS) method was developed for the simultaneous determination of 12 bioactive components in Gegen-Qinlian decoction. After optimization, the extraction conditions were set as follows: 3.0 mL of aqueous sample solution (pH 5.86) was extracted with 2 mL of acetonitrile at 235°C for 35 min. The separated acetonitrile phase was diluted 10-fold with water before UPLC-MS-MS analysis. Separation was performed on a Waters ACQUITY UPLC®BEH C18 column (2.1 × 100 mm i.d., 1.7 μm) with ammonium formate buffer solution (20 mmol L-1, pH 3.2, adjusted by formic acid) and acetonitrile as mobile phase with gradient elution. Twelve target components could be separated within 10 min and quantified in multiple reaction monitoring mode, both positive and negative ionization modes were employed. Limits of detection were in the range of 0.0003- 0.0451 μg mL-1. Relative standard deviation values for intra- and interday precision were <2.71 and 8.94%, respectively. The established method provides a simple and effective framework for the quality control of Gegen-Qinlian decoction and related traditional Chinese medicinal preparations. [ABSTRACT FROM AUTHOR]

Background: Surgery plays an important role in the management of endometrial cancer at all stages, particularly early clinical stage. There are still many unanswered questions regarding optimal surgical management of endometrial cancer, particularly regarding which patients should undergo lymphadenectomy. The aim of this study was to evaluate the role of preoperative cancer antigen 125 (CA125) serum levels for surgical management in endometrial cancer patients. Methods: A total of 995 patients with endometrial cancer, according to inclusion criteria of a preoperative serum level of CA125, were selected. The association between clinicopathological factors and CA125 were analyzed. Receiver operating characteristic (ROC) curve was used to evaluate the role of preoperative serum CA125 in predicting lymph node metastasis, adnexal involement, cervical stromal invasion in all patients, especially patients with clinical stage I. Survival analyses were also performed according to the four groups of preoperative CA125 serum levels. Results: Elevated CA125 level was significantly associated with all clinicopathological parameters, including age and menopause, but not histology type. ROC curve analysis results showed the CA125 serum level of 25 U/mL was the best cutoff to predict the lymph node metastasis. It was with 78% of sensitivity, 78% of specificity, 77.6% of false positive rate, 2.3% of false negative rate in all patients. In patients with clinical stage I, it was with 71.7% of sensitivity, 77.6% of specificity, 83.3% of false positive rate, 2.2% of false negative rate. The best cutoff to evaluate adnexal involement in patients with clinical stage I was 30U/ml, with 81% sensitivity, and 78.4% specificity. Survival analysis revealed CA125, FIGO stage, histology grade, and positive peritoneal cytology as independent prognostic factors of endometrial cancer. Conclusion: Preoperative serum CA125 is an important predictor for patients with endometrial cancer and it should be taken into consideration when surgical management is determined, especially if a lymphadenectomy should be undertaken in patients with clinical stage I. [ABSTRACT FROM AUTHOR]

Aim of the study: In the absence of effective therapeutic strategies, cervical carcinoma continues to be second on the list of mortality rates of malignant tumours found in women. We investigated the effects of β-sitosterol, a natural product isolated from traditional Chinese herbs, on Caski and HeLa cervical carcinoma cells. Material and methods: Morphological changes were examined by light microscopy. Ultrastructures of Caski and HeLa cells treated with 20 µmol/l β-sitosterol were documented by scanning electron microscopy and transmission electron microscopy. Changes in mRNA and protein expression were quanti- fied respectively using Real-Time qPCR and western blot methods. Results: Treatment of Caski and HeLa cells with β-sitosterol resulted in reduced expression of PCNA, indicative of an inhibitory effect on cell proliferation. This was associated with increased p53 mRNA levels and decreased amounts of HPV E6 transcripts. Expression of p53 and HPV E6 proteins followed a similar trend as that observed for the corresponding transcripts. Caski and HeLa cells treated with β-sitosterol exhibited loss of cell surface microvilli, increased electron density of cell membrane, and decreased organelles. Conclusions: In conclusion, treatment of Caski and HeLa cells with β-sitosterol significantly suppressed the expression of genes involved in cell proliferation and oncogenic transformation, while augmenting the expression of genes involved in apoptosis and tumour suppression. Ultrastuctural characterisation of Caski and HeLa cells treated with β-sitosterol further confirmed the anti-proliferative and anti-cancer activity of this natural product isolated from traditional Chinese herbs. [ABSTRACT FROM AUTHOR]

Background: While HPV infection is the main cause of cervical cancer, genetic susceptibility to HPV infection is not well understood. Tumor necrosis factor alpha (TNF-alpha), involved in the defense against HPV infection, plays an important role in cervical cancer progression and regression. The aim of this study was to investigate the relationship between the TNF- alpha rs1800629 polymorphism and risk of HPV infection or cervical cancer. Methods: Three groups were involved in this study of Chinese women. Group 1 consisted of 285 high risk HPV positive cervical cancer patients, Group 2, 225 high risk HPV positive patients without cervical cancer, and Group 3, 318 HPV negative women with no cervical cancer. Blood samples were obtained from all patients and genotyped by PCR-RLFP. Fifty randomly selected samples were further sequenced. Results: The allele and genotype distributions of the TNF-alpha rs1800629 polymorphism were not significantly different between each of the groups (P>0.05). There are no significant relationship between rs1800629 polymorphism and high risk HPV infection (OR = 0.649, 95% CI: 0.253-1.670, P = 0.371), cervical cancer (OR = 0.993, 95% CI: 0.376-2.618, P = 0.988), or cervical cancer with HPV infection (OR = 0.663, 95% CI: 0.250-1.758, P = 0.409). Conclusions: We demonstrated that there is no association between TNF rs1800629 polymorphism and the HPV infection, or cervical cancer with HPV infection. [ABSTRACT FROM AUTHOR]

Wheat, rice, and maize collectively provide the bulk of energy and protein to the human diet worldwide. Over the last seven decades, China increased 5.8-fold the aggregated production of the three cereals, but further gains are required to meet the predicted demand of 776 Mt. by 2030. Here, we report the rate of genetic gain in yield of 0.65% year-1 for wheat between 1937 and 2012, 0.69% year-1 for rice between 1934 and 2011, and 0.77% year-1 for maize between 1950 and 2014 in China. National farm yield in the same periods increased 1.38% year-1 for wheat, 1.15% year-1 for rice and 1.32% year-1 for maize. Hence, genetic improvement (G) accounted for 47%-60% of the increase in farm yield, with improved management (M) and G × M interaction accounting for the balance. The absolute rate of genetic gain (kg ha-1 year-1) was proportional to environmental potential. Genetic gain in nitrogen utilization efficiency (yield per unit nitrogen uptake) was significant for all three crops and was achieved at the expense in grain N concentration that decreased at 0.19% year-1 in rice and 0.47% year-1 in maize. To guarantee future food security and environmental sustainability, breeding and agronomic efforts need to increase the rate of gain in yield and revert the declining trends in grain protein by improving nitrogen uptake and nitrogen harvest index. [ABSTRACT FROM AUTHOR]

• Nanosized precipitates can be characterized by the atomic force microscope. • Vibration and electrolytic polishing all can be used for AFM sample preparation. • Precipitate fraction can be calculated based on AFM images by using IPP software. Precipitate was typically characterized by the transmission electron microscopy (TEM) or scanning electron microscopy (SEM) to analyze the relationship between material property and precipitate fraction. However, these procedures tend to be are always time consuming because of the complicated sample preparation process involved and the observation course. Particularly, sometime the precipitate can't be highlighted solely from the images easily. In this study, atomic force microscopy (ATM) was performed to characterize the morphology and fraction of the precipitate in this paper. Five kinds of materials were selected and prepared by electropolishing or vibration polishing method to display the precipitates on the sample surface. The experiment results prove that different types of precipitates with higher values for height on a sample surface could be clearly observed by AFM, and the images quality is highly relied upon the surface quality. The precipitate fraction can be calculated using the AFM micrographs and image photo post treated software (IPP). It was turned out that AFM is found suitable for observation almost all kinds of precipitates, and the precipitates can be easily separated from the images. Not only vibration polishing but also electrolytic polishing could offer a smooth surface for observing nanosized precipitates by AFM. A comparison of the fraction result obtained by AFM and precise microchemical analysis proves that the fraction measurement result obtained by AFM is acceptable. [ABSTRACT FROM AUTHOR]

Objective: To determine the adult reference intervals for the Siemens IgG subclass reagents. Methods: 636 blood samples of healthy adults were analyzed to determine the level of IgG subclass using the reagents of Siemens immunonephelometric assay with molecular biology kits. Results: IgGSc reference intervals were as follows: IgG1 4.45-9.76 g/L, IgG2 2.07-8.57 g/L, IgG3 0.08-0.80 g/L and IgG4 0.05-1.54 g/L. There was an excellent correlation between the total IgG and the sum of the IgG subclasses. No significant gender and age differences were observed. Conclusions: Our data provide the missing reference intervals and enable the use of the nephelometric IgG subclass reagents in Chinese. The study can offer reference on clinic diagnose. [ABSTRACT FROM AUTHOR]

This study assessed changes in nitrogen (N) and phosphorus (P) uptake and partitioning in response to selection for yield in milestone varieties of Chinese winter wheat (Triticum aestivum L.). We established a factorial trial combining 11 nutrient–water regimes with three (2013–14) and five (2014–15) varieties released from 1970 to 2005. Grain yield increased at a rate of 0.46% year–1, with no apparent increase in the uptake of nutrients. Nitrogen harvest index did not change, and P harvest index increased at a rate of 0.15% year–1. Consequently, yield per unit N uptake and yield per unit P uptake increased at similar rates (0.4% year–1) at the expense of nutrient concentration in grain, which declined at a rate of 0.47% year–1 for N and 0.31% year–1 for P. No trends in N nutrition index were found. Selection for yield in wheat increased the yield per unit nutrient uptake at the expense of grain nutrient concentration. Further gains in yield need to be matched by increasing N uptake to maintain grain protein. Dilution of P in grain needs to be considered in terms of the putatively undesirable role of phytate for human nutrition, and the need for P reserves in seed for crop establishment. [ABSTRACT FROM AUTHOR]

The symbiosis between leguminous plants and soil rhizobia culminates in the formation of nitrogen-fixing organs called nodules that support plant growth. Two Medicago truncatula Tnt1-insertion mutants were identified that produced small nodules, which were unable to fix nitrogen effectively due to ineffective rhizobial colonization. The gene underlying this phenotype was found to encode a protein containing a putative membrane-localized domain of unknown function (DUF21) and a cystathionine-β-synthase domain. The cbs1 mutants had defective infection threads that were sometimes devoid of rhizobia and formed small nodules with greatly reduced numbers of symbiosomes. We studied the expression of the gene, designated M. truncatula Cystathionine-β-Synthase-like1 (MtCBS1), using a promoter-β-glucuronidase gene fusion, which revealed expression in infected root hair cells, developing nodules, and in the invasion zone of mature nodules. An MtCBS1-GFP fusion protein localized itself to the infection thread and symbiosomes. Nodulation factor-induced Ca2+ responses were observed in the cbs1 mutant, indicating that MtCBS1 acts downstream of nodulation factor signaling. MtCBS1 expression occurred exclusively during Medicago-rhizobium symbiosis. Induction of MtCBS1 expression during symbiosis was found to be dependent on Nodule Inception (NIN), a key transcription factor that controls both rhizobial infection and nodule organogenesis. Interestingly, the closest homolog of MtCBS1, MtCBS2, was specifically induced in mycorrhizal roots, suggesting common infection mechanisms in nodulation and mycorrhization. Related proteins in Arabidopsis have been implicated in cell wall maturation, suggesting a potential role for CBS1 in the formation of the infection thread wall. [ABSTRACT FROM AUTHOR]

Background: Monoclonal antibodies (mAb) that block programmed death (PD)-1 signaling pathway hold great potential as a novel cancer immunotherapy. Recent evidence suggests that combining with conventional, targeted or other immunotherapies, these mAb can induce synergistic antitumor responses. In this study, we investigated whether Trabectedin (ET-743), a novel anticancer agent currently used for treating relapsed ovarian cancer, can synergize with anti (α)-PD-1 mAb to increase antitumor activity in the murine ID8 ovarian cancer model. Methods: Mice with established peritoneal ID8 tumor were treated with either single or combined Trabectedin and α-PD-1 mAb, their overall survival was recorded; tumor-associated immune cells and immune gene expression in tumors from treated mice were analyzed by flow cytometry and quantitative RT-PCR, respectively, and antigenspecific immunity of effector CD8+ T cells was evaluated by ELISA and cytotoxicity assay. In addition, the effect of Trabectedin on tumoral PD-L1 expression was analyzed by both flow cytometry and immunofluorescence staining. Results: Though single treatment showed a modest antitumor effect in mice bearing 10-day-established ID8 tumor, combined Trabectedin and α-PD-1 mAb treatment induced a strong antitumor immune response, leading to a significant tumor regression with half of mice tumor-free 90 days after tumor inoculation. Mechanistic investigation revealed that combination treatment induces a systemic tumor-specific immunity with an indispensable role of both CD4+ and CD8+ T cells, and effector CD8+ T cells exhibited the antigen-specific cytokine secretion and cytotoxicity upon tumor antigen stimulation; additionally, combination treatment increased the IFN-γ-producing effector T cells and decreased the immunosuppressive cells in peritoneal cavity; accordingly, it enhanced the expression of Th1-associated immune-stimulating genes while reducing the transcription of regulatory/suppressive immune genes, reshaping tumor microenvironment from a immunosuppressive to a stimulatory state. Finally, in vivo Trabectedin treatment has been shown to induce IFN-γ-dependent PD-L1 expression within tumor, possibly constituting a mechanistic basis for its synergistic antitumor effect with α-PD-1 mAb therapy. Conclusion: This study provides the evidence that α-PD-1 mAb can produce a synergistic antitumor efficacy when combined with Trabectedin, a clinically available anticancer agent, supporting a direct translation of this combination strategy in clinic for the treatment of ovarian cancer. [ABSTRACT FROM AUTHOR]

In this paper, graphene, a new sorbent material, was synthesized and used for solid-phase extraction (SPE) of the six carbamate pesticides (pirimicarb, baygon, carbaryl, isoprocarb, baycarb and diethofencarb) in environmental water samples. The target analytes can be extracted on the graphene-packed SPE cartridge, and then eluted with acetone. The eluate was collected and dried by high purity nitrogen gas at room temperature. 1mL of 20% (v/v) acetonitrile aqueous solution was used to redissolve the residue. The final sample solution was analyzed by ultra performance liquid chromatography-tandem quadrupole mass spectrometry (UPLC-MS/MS) system. Under optimum conditions, good linearity was obtained for the carbamates with correlation coefficient in the range of 0.9992-0.9998. The limits of detection (S/N=3) for the six carbamate pesticides were in the range of 0.5-6.9ngL-1. Relative standard deviations (RSD) for five replicate determinations were below 5.54%. RSD values for cartridge-to-cartridge precision (n=7) were in the range of 1.27-8.13%. After proper regeneration, the graphene-packed SPE cartridge could be re-used over 100 times for standard solution without significant loss of performance. The enrichment factors for the target analytes were in the range of 34.2-51.7. The established method has been successfully applied to the determination of carbamate pesticide residues in environmental water samples such as river water, well water and lake water. [ABSTRACT FROM AUTHOR]

We investigated the significance of high- mobility group box1 (HMGB1) and T-cell-mediated immunity and prognostic value in cervical cancer. HMGB1, forkhead/winged helix transcription factor p3 (Foxp3), IL-2, and IL-10 protein expression was analyzed in 100 cervical tissue samples including cervical cancer, cervical intraepithelial neoplasia (CIN), and healthy control samples using immunohistochemistry. Serum squamous cell carcinoma antigen (SCC-Ag) was immunoradiometrically measured in 32 serum samples from 37 cases of squamous cervical cancer. HMGB1 and SCC-Ag were then correlated to clinicopathological characteristics. HMGB1 expression tends to increase as cervical cancer progresses and it was found to be significantly correlated to FIGO stage and lymph node metastasis. These findings suggest that HMGB1 may be a useful prognostic indicator of cervical carcinoma. In addition, there were significant positive relationships between HMGB1 and FOXP3 or IL-10 expression (both p < 0.05). In contrast, HMGB1 and IL-2 expression was negatively correlated (p < 0.05). HMGB1 expression may activate Tregs or facilitate Th2 polarization to promote immune evasion of cervical cancer. Elevated HMGB1 protein in cervical carcinoma samples was associated with a high recurrence of HPV infection in univariate analysis (p < 0.05). HMGB1 expression and levels of SCC-Ag were directly correlated in SCC (p < 0.05). Thus, HMGB1 may be a useful biomarker for patient prognosis and cervical cancer prediction and treatment. [ABSTRACT FROM AUTHOR]

Objective: This study investigates the expression of Lewis y antigen, integrin αv, β3 in epithelial ovarian cancer tissues. We further evaluate the relationship between their expression and chemotherapy resistance of ovarian cancer and its possible clinical significance. Methods: Tissues of 92 patients with ovarian cancer meeting the inclusion criteria with complete follow-up data were enrolled and divided into chemotherapy resistant group and sensitive group. The expression and relationship of Lewis y antigen and integrin αv, β3 are assessed in paraffin sections using immunohistochemistry and doublelabeling immunofluorescence method. Multivariate logistic regression analysis was used to investigate the relationship between age, clinical stage, differentiation, histologic subtype, Lewis y antigen and integrin αv, β3 expression in ovarian cancer patients. Results: The expression rates of Lewis y antigen and integrin αv in the resistant group, significantly higher than the rates found in the sensitive group (p <0.05). Multivariate analysis showed that the expression of Lewis y antigen, integrin αv and ovarian cancer's clinical stage were independent, drug resistance-related risk factors. The expression levels of Lewis y antigen and integrin αv, β3 were positively correlated with each other. Conclusions: A close correlation between Lewis y antigen, integrin αv, β3 and ovarian cancer was observed. Lewis y antigen can influence the biological behavior of a tumor cell as an important composition of integrin αv, β3 by some signal pathway. And the expression of Lewis y antigen, integrin αv and ovarian cancer's clinical stage are both independent, drug resistance-related risk factors. [ABSTRACT FROM AUTHOR]

Resistin is a novel hormone that is secreted by human adipocytes and mononuclear cells and is associated with obesity, insulin resistance and inflammation. Recently, resistin has been postulated to play a role in angiogenesis. Here, we investigated the hypothesis that resistin regulates ovary carcinoma production of vascular endothelial growth factor (VEGF) and the angiogenic processes. We found that in human ovarian epithelial carcinoma cells (HO-8910), resistin (10-150 ng/mL) enhanced both VEGF protein and mRNA expression in a time- and concentration-dependent manner, as well as promoter activity. Furthermore, resistin enhanced DNA-binding activity of Sp1 with VEGF promoter in a PI3K/Akt-dependent manner. PI3K/Akt activated by resistin led to increasing interaction with Sp1, triggering a progressive phosphorylation of Sp1 on Thr453 and Thr739, resulting in the upregulation of VEGF expression. In an in vitro angiogenesis system for endothelial cells (EA.hy926) co-cultured with HO-8910 cells, we observed that the addition of resistin stimulated endothelial cell tube formation, which could be abolished by VEGF neutralizing antibody. Our findings suggest that the PI3K/Akt-Sp1 pathway is involved in resistin-induced VEGF expression in HO-8910 cells and indicates that antiangiogenesis therapy may be beneficial treatment against ovarian epithelial carcinoma, especially in obese patients. [ABSTRACT FROM AUTHOR]

Objective: To measure Lewis y and integrin α5β1 expression in epithelial ovarian carcinoma and to correlate the levels of these molecules with ovarian carcinoma chemotherapy and prognosis. Methods: The study population included 34 ovarian carcinoma patients with chemotherapeutic drug-resistance, six partially drug-sensitive cases, and 52 drug-sensitive cases (92 total). Immunochemistry was used to determine expression of Lewis y antigen and integrin α5β1 in ovarian carcinoma tissues, and correlation of these molecules with chemotherapy resistance was further investigated, Multi-factor logistic regression analysis was applied to investigate: age, surgical stage, grade, subtype of patient cases, metastasis of lymph nodes, residual tumor size, expression levels of Lewis y antigen and integrin α5β1 correlation with ovarian carcinoma chemotherapy resistance. Results: The expression rates of Lewis y antigen and integrins α5 and β1 were significantly greater in the drug-resistant group (91.17%, 85.29%, 88.24%) than the partially sensitive (50.00%, 33.33%, 50.00%) or sensitive groups (61.54%, 57.69%, 55.77%). Binary logistic regression analysis revealed that surgical stage, residual tumor size, and expression of integrin α5 and Lewis y in ovarian carcinoma tissues were independent risk factors for chemotherapeutic drug resistance. Conclusions: Overexpression of Lewis y and integrin α5 are strong risk factors for chemotherapeutic drug resistance in ovarian carcinoma patients. [ABSTRACT FROM AUTHOR]

Objective: To investigate the effect of Lewis y overexpression on the expression of proliferation-related factors in ovarian cancer cells. Methods: mRNA levels of cyclins, CDKs, and CKIs were measured in cells before and after transfection with the α1,2-fucosyltransferase gene by real-time PCR, and protein levels of cyclins, CDKs and CKIs were determined in cells before and after gene transfection by Western blot. Results: Lewis y overexpression led to an increase in both mRNA and protein expression levels of cyclin A, cyclin D1 and cyclin E in ovarian cancer cells, decrease in both mRNA and protein expression levels of p16 and p21, and decrease of p27 at only the protein expression level without change in its mRNA level. There were no differences in proteins and the mRNA levels of CDK2, CDK4 and CDK6 before and after gene transfection. Anti-Lewis y antibody, ERK and PI3K pathway inhibitors PD98059 and LY294002 reduced the difference in cyclin and CKI expression caused by Lewis y overexpression. Conclusion: Lewis y regulates the expression of cell cycle-related factors through ERK/MAPK and PI3K/Akt signaling pathways to promote cell proliferation. [ABSTRACT FROM AUTHOR]

Epithelial carcinomas of the ovary exhibit the highest mortality rate among gynecologic malignancies. Studies found that the metabolism of glycolipids or carbohydrates is associated with acquirement of anticancer drug-resistance by cancer cells. This study was to characterize possible involvement of Lewis Y (LeY) antigen in the drug-resistance of cancer cells. We transfected the α1,2-fucosyltransferase gene into human ovarian carcinoma-derived RMG-1 cells and established RMG-1-hFUT cells with enhanced expression of LeY. We determined the effects of docetaxel on the survival of cells by MTT assaying and observed the apoptosis of cells in the presence of docetaxel by flow cytometric analysis and by transmission electron microscopy. Plasma membranes and intracellular granules in RMG-1-hFUT cells were stained with anti-LeY antibody, the intensity of the staining was higher than that in control cells. The RMG-1-hFUT cells exhibited higher resistance to docetaxel than the control cells with regard to the docetaxel concentration and time course. After treatment with 10 µg/mL docetaxel for 72 h, the control cells, but not RMG-1-hFUT, contained abundant positively stained cell debris due to disintegration of the cytoskeleton. On transmission electron microscopy, although the control cells treated with docetaxel as above showed the following morphology, i.e., absence of villi, cells shrunken in size, pyknosis, agglutinated chromatin and cell buds containing nuclei in the process of apoptosis, the RMG-1-hFUT cells showed only agglutinated chromatin and vacuoles in the cytoplasm. In summary, cells with enhanced expression of LeY were shown to acquire docetaxel-resistance, indicating the possible involvement of glycoconjugates in the drug-resistance. [ABSTRACT FROM AUTHOR]

Objective: This study aimed to measure and correlate the expression of nsulin-like growth factor receptor-1 (IGF-1R) and the Lewis(y) antigen in ovarian cancer cell lines and tissue samples. Methods: Reverse transcriptase PCR (RT-PCR), Western blotting, immunoprecipitation, immunohistochemistry, and immunofluorescence double-labeling techniques were applied to detect and measure the expression of Lewis(y) and IGF-1R. Results: In &agr;1,2-fucosyltransferase (&agr;1,2-FT)-transfected cells, IGF-1R expression was significantly upregulated compared with cells that do not overexpress R1,2-FT (P < 0.05). The amount of Lewis(y) expressed on IGF-1R increased 1.81-fold in &agr;1,2-FT-overexpressing cells (P < 0.05), but the ratio of Lewis(y) expressed on IGF-1R to total IGF-1R was unaltered between two cells (P > 0.05). In malignant epithelial ovarian tumors, the positivity rates of Lewis(y) and IGF-1R detection were 88.3% and 93.33%, respectively, which is higher than the positivity rates in marginal (60.00% and 63.33%, all P < 0.05), benign (33.00% and 53.33%, all P < 0.01), and normal (0% and 40%, all P < 0.01) ovarian samples. No correlations were detected in positivity rates of Lewis(y) or IGF-1R expression with respect to clinicopathological parameters in ovarian cancers (all P > 0.05). Both IGF-1R and Lewis(y) were highly expressed in ovarian cancer tissues, and their expression levels were positively correlated (P < 0.05). Conclusion: Overexpression of Lewis(y) results in overexpression of IGF-1R. Both IGF-1R and Lewis(y) are associated with the occurrence and development of ovarian cancers. [ABSTRACT FROM AUTHOR]

Objective: To detect the expression and clinical significances of Lewis y antigen and integrin αv, β3 in epithelial ovarian tumors, and to explore the expression correlation between Lewis y antigen and integrin αv, β3. Methods: Immunohistochemical staining was performed in 95 cases of epithelial ovarian cancer, 37 cases of borderline tumors, 20 cases of benign tumors, and 20 cases of normal ovarian tissue, for the detection of Lewis y antigen and integrin αv, β3 expressions, and to analyze the relationship between Lewis y antigen and integrin, and the relationship between clinical and pathological parameters of ovarian cancer. In addition, immunofluorescence double labeling was utilized to detect the expression correlation between Lewis y antigen and integrin αv, β3 in ovarian cancer. Results: In epithelial ovarian tumors, the expression rate of Lewis y antigen was 81.05%, significantly higher than that of borderline (51.53%) (P < 0.05) and benign (25%) (P < 0.01) tumors, and normal ovarian tissues (0) (P < 0.01). The expression rate of integrin αv, β3 in malignant epithelial ovarian tumors was 78.95% and 82.11%, respectively, significantly higher than that of the borderline (45.94%, 40.54%) (both P < 0.05), benign group (10.00%, 15.00%) (both P < 0.01) and normal ovary group (5%, 15%) (both P < 0.01). Conclusions: Lewis y and integrins αv, β3 are relevant to pelvic and abdominal diffusion and metastasis of ovarian cancer cells, suggesting that these two molecules mediate a boosting function for tumor metastasis. [ABSTRACT FROM AUTHOR]

Background: This study aimed to investigate the molecular structural relationship between cell adhesive molecule CD44 and Lewis y antigen, and determine the effects of Lewis y antigen on CD44-mediated adhesion and spreading of ovarian cancer cell line RMG-I and the Lewis y antigen-overexpressed cell line RMG-I-H. Methods: The expression of CD44 in RMG-I and RMG-I-H cells before and after treatment of Lewis y monoclonal antibody was detected by immunocytochemistry; the expression of Lewis y antigen and CD44 was detected by Western Blot. The structural relationship between Lewis y antigen and CD44 was determined by immunoprecipitation and confocal laser scanning microscopy. The adhesion and spreading of RMG-I and RMG-I-H cells on hyaluronic acid (HA) were observed. The expression of CD44 mRNA in RMG-I and RMG-I-H cells was detected by real-time RT-PCR. Results: Immunocytochemistry revealed that the expression of CD44 was significantly higher in RMG-I-H cells than in RMG-I cells (P < 0.01), and its expression in both cell lines was significantly decreased after treatment of Lewis y monoclonal antibody (both P < 0.01). Western Blot confirmed that the content of CD44 in RMG-I-H cells was 1.46 times of that in RMG-I cells. The co-location of Lewis y antigen and CD44 was confirmed by co-immunoprecipitation. The co-expression of CD44 and Lewis y antigen in RMG-I-H cells was 2.24 times of that in RMG-I cells. The adhesion and spreading of RMG-I-H cells on HA were significantly enhanced as compared to those of RMG-I cells (P < 0.01), and this enhancement was inhibited by Lewis y monoclonal antibody (P < 0.01). The mRNA level of CD44 in both cell lines was similar (P > 0.05). Conclusion: Lewis y antigen strengthens CD44-mediated adhesion and spreading of ovarian cancer cells. [ABSTRACT FROM AUTHOR]

Lewis (y) antigen is a difucosylated oligosaccharide present on the plasma membrane, and its overexpression is frequently found in human cancers and has been shown to be associated with poor prognosis. Our previous studies have shown that Lewis (y) antigen plays a positive role in the process of invasion and metastasis of ovarian cancer cells. However, the mechanisms by which Lewis (y) antigen enhances the invasion and tumor metastasis are still unknown. In this study, we established a stable cell line constitutively expressing Lewis (y) antigen (RMG-1-hFUT) by transfecting the cDNA encoding part of the human α1,2-fucosyltransferase (α1,2-FUT) gene into the ovarian cancer cell line RMG-1, and investigated whether Lewis (y) antigen regulates the expression of matrix metalloproteinase-2 (MMP-2) and MMP-9, and tissue inhibitors of metalloproteinases (TIMP-1) and TIMP-2. We found that RMG-1-hFUT cells exhibited higher invasive capacities than their control cells. In addition, expression of TIMP-1 and TIMP-2 was down-regulated and expression of MMP-2 and MMP-9 was up-regulated. Anti-Lewis (y) antigen antibody treatment significantly reversed the expression of TIMP-1, TIMP-2, MMP-2 and MMP-9. Taken together, we provide the first evidence that down-regulation of TIMP-1 and TIMP-2 and up-regulation of MMP-2 and MMP-9 represents one of the mechanisms by which Lewis (y) antigen promotes cell invasion. [ABSTRACT FROM AUTHOR]

Lewis y (LeY) antigen is a difucosylated oligosaccharide carried by glycoconjugates on the cell surface. Overexpression of LeY is frequently observed in epithelial-derived cancers and has been correlated to the pathological staging and prognosis. However, the effects of LeY on ovarian cancer are not yet clear. Previously, we transfected the ovarian cancer cell line RMG-I with the α1,2-fucosyltransferase gene to obtain stable transfectants, RMG-I-H, that highly express LeY. In the present study, we examined the proliferation, tumorigenesis, adhesion and invasion of the cell lines with treatment of LeY monoclonal antibody (mAb). Additionally, we examined the expression of TGF-β1, VEGF and b-FGF in xenograft tumors. The results showed that the proliferation and adhesion in vitro were significantly inhibited by treatment of RMG-I-H cells with LeY mAb. When subcutaneously inoculated in nude mice, RMG-I-H cells produced large tumors, while mock-transfected cells RMG-I-C and the parental cells RMG-I produced small tumors. Moreover, the tumor formation by RMG-I-H cells was inhibited by preincubating the cells with LeY mAb. Notably, the expression of TGFβ1, VEGF and b-FGF all increased in RMG-I-H cells. In conclusion, LeY plays an important role in promoting cell proliferation, tumorigenecity and adhesion, and these effects may be related to increased levels of growth factors. The LeY antibody shows potential application in the treatment of LeY-positive tumors. [ABSTRACT FROM AUTHOR]

Aim: The expression of Dkk-3, as a number of the Dkk family was different in different cancer. Because Dkk-3 encodes a secreted protein, we investigated whether the Dkk-3 protein is secreted into the sera of patients with gynecological cancer. Material & Methods: The levels of Dkk-3 protein were assessed by enzyme-linked immunosorbent assay in the sera of 104 patients with gynecological cancer: 36 with ovarian, 40 with cervical and 28 with endometrial cancers. Results: The serum levels of Dkk-3 protein in patients with ovarian cancer [25.54 (7.99) pg/mL] was lower than in normal individuals [42.08 (14.89) pg/mL] ( P = 0.000). But serum levels of Dkk-3 protein in patients with cervical [166.39 (300.68) pg/mL] ( P = 0.013) and endometrial cancers [73.64 (23.36) pg/mL] ( P = 0.000) were higher than in normal individuals. The serum levels of Dkk-3 protein were associated with clinical stage in patients with cervical and endometrial cancers. In patients with ovarian cancer, the serum levels of Dkk-3 protein were associated with lymphatic metastasis. In patients with cervical cancer, the serum levels of Dkk-3 protein were associated with tumor diameters. Conclusion: Dkk-3 protein detection using enzyme-linked immunosorbent assay as molecular markers can contribute to detection and diagnosis of gynecological cancer, especially for ovarian cancer and endometrial cancer. [ABSTRACT FROM AUTHOR]

Microbial biomass phosphorus (MBP) and its relationships with other biological and chemical properties were studied on loess soil with an 11-year long-term fertilization experiment. The results indicated that inorganic fertilizers (F) improved soil microbial biomass carbon (MBC), nitrogen (MBN), and MBP levels and F plus maize stalk (SNPK) improved MBC and MBN. Manuring markedly increased soil MBC, MBN, and MBP levels. Fertilization decreased the ratios of MBC/MBN, MBC/MBP, and MBN/MBP. Microbial biomass phosphorus was positively and linearly correlated with MBC, MBN, organic carbon (SOC), total phosphorus (TP), water-soluble P, and Olsen P but negatively correlated with soil pH. Microbial biomass phosphorus constituted 2% of TP on control (CK) and inorganic fertilizer treatments and 12% on manure plots. Microbial biomass phosphorus to Olsen P ratios were 50% on CK, F, and SNPK and 80% on manure treatments. Measurements of MBP in soil containing high Olsen P were subject to analytical problems of unknown reasons. [ABSTRACT FROM AUTHOR]

Background: To determine the expression of bone morphogenetic protein-2 (BMP-2) and its receptors BMPRIA, BMPRIB, and BMPRII in epithelial ovarian cancer (EOC) and to analyze their influence on the prognosis of ovarian cancer patients. Methods: Semi-quantitative RT-PCR and western blot were applied to detect the expression of BMP-2 and its receptors BMPRIA, BMPRIB, and BMPRII in EOC, benign ovarian tumors, and normal ovarian tissue at the mRNA and protein levels. Immunohistochemistry was used to determine the expression of BMP-2 and its receptors in 100 patients with EOC to analyze their influence on the five-year survival rate and survival time of ovarian cancer patients. Results: (1) The mRNA and protein expression levels of BMP-2, BMPRIB, and BMPRII in ovarian cancer tissue were remarkably lower than those in benign ovarian tumors and normal ovarian tissue, while no significant differences in BMPRIA expression level was found among the three kinds of tissues. (2) The five-year survival rate and the average survival time after surgery of EOC patients with positive expression of BMP-2, BMPRIB, and BMPRII were remarkably higher than those of patients with negative expression of BMP-2, BMPRIB, and BMPRII. BMPRIA expression was not associated with the five-year survival rate or with the average survival time of ovarian cancer patients. Conclusions: BMP-2, BMPRIB, and BMPRII exhibited low expression in EOC tissue, and variation or loss of expression may indicate poor prognosis for ovarian cancer patients. [ABSTRACT FROM AUTHOR]