Aquaporin 7 is upregulated through the PI3K-Akt pathway and modulates decidualisation of endometrial stromal cells.

CONTEXT: Aquaporin 7 (AQP7) is selectively expressed in decidualised endometrial stromal cells (ESCs) of mice surrounding the embryonic implantation sites. However, the roles of AQP7 and the underlying mechanism that regulates AQP7 expression in endometrial decidualisation after implantation are still unclear. AIMS: This study aimed to investigate the role of the PI3K-Akt pathway in regulating the expression of AQP7 in ESCs and decidualisation. METHODS: Primary ESCs of pregnant mice were isolated to establish in vitro decidualisation models. PI3K inhibitor LY294002 was added to the decidualisation models, then AQP7 expression, changes in decidualised ESC morphology and expression of decidualisation marker molecules were examined. KEY RESULTS: AQP7 knockdown reduced the proliferation and differentiation of ESCs with in vitro induced decidualisation. Furthermore, when the activity of PI3K was inhibited by LY294002, the expression of AQP7 in decidualised ESCs was decreased and both the proliferation and differentiation of ESCs were significantly reduced. CONCLUSIONS: This indicates that AQP7 is a key molecule involved in endometrial decidualisation and the expression of AQP7 is upregulated through activation of the PI3K-Akt pathways, which promotes the proliferation and differentiation of the ESCs, thus affecting occurrence of decidualisation. IMPLICATIONS: This study may provide a new biomarker for the diagnosis of infertility and a new drug target for the prevention and treatment of infertility.

Bi-specific T1 positive-contrast-enhanced magnetic resonance imaging molecular probe for hepatocellular carcinoma in an orthotopic mouse model.

BACKGROUND: Hepatocellular carcinoma (HCC) is the second leading cause of cancer-related mortality. HCC-targeted magnetic resonance imaging (MRI) is an effective noninvasive diagnostic method that involves targeting clinically-related HCC biomarkers, such as alpha-fetoprotein (AFP) or glypican-3 (GPC3), with iron oxide nanoparticles. However, in vivo studies of HCC-targeted MRI utilize single-target iron oxide nanoprobes as negative (T2) contrast agents, which might weaken their future clinical applications due to tumor heterogeneity and negative MRI contrast. Ultra-small superparamagnetic iron oxide (USPIO) nanoparticles (approximately 5 nm) are potential optimal positive (T1) contrast agents. We previously verified the efficiency of AFP/GPC3-double-antibody-labeled iron oxide MR molecular probe in vitro. AIM: To validate the effectiveness of a bi-specific probe in vivo for enhancing T1-weighted positive contrast to diagnose the early-stage HCC. METHODS: The single- and double-antibody-conjugated 5-nm USPIO probes, including anti-AFP-USPIO (UA), anti-GPC3-USPIO (UG), and anti-AFP-USPIO-anti-GPC3 (UAG), were synthesized. T1- and T2-weighted MRI were performed on day 10 after establishment of the orthotopic HCC mouse model. Following intravenous injection of U, UA, UG, and UAG probes, T1- and T2-weighted images were obtained at 12, 12, and 32 h post-injection. At the end of scanning, mice were euthanized, and a histologic analysis was performed on tumor samples. RESULTS: T1- and T2-weighted MRI showed that absolute tumor-to-background ratios in UAG-treated HCC mice peaked at 24 h post-injection, with the T1- and T2-weighted signals increasing by 46.7% and decreasing by 11.1%, respectively, relative to pre-injection levels. Additionally, T1-weighted contrast in the UAG-treated group at 24 h post-injection was enhanced 1.52-, 2.64-, and 4.38-fold compared to those observed for single-targeted anti-GPC3-USPIO, anti-AFP-USPIO, and non-targeted USPIO probes, respectively. Comparison of U-, UA-, UG-, and UAG-treated tumor sections revealed that UAG-treated mice exhibited increased stained regions compared to those observed in UG- or UA-treated mice. CONCLUSION: The bi-specific T1-positive contrast-enhanced MRI probe (UAG) for HCC demonstrated increased specificity and sensitivity to diagnose early-stage HCC irrespective of tumor size and/or heterogeneity.

Prognostic analysis and establishment of a nomogram in patients with myoepithelial carcinoma of the salivary gland: A population-based study.

BACKGROUND: Currently, the clinicopathological characteristics and prognosis of myoepithelial carcinoma of salivary gland (MC-SG) have not been defined well. The present study aimed to describe the clinicopathological characteristics and prognosis of MC-SG patients. METHODS: The Surveillance, Epidemiology, and End Results database was searched for all patients diagnosed with MC-SG between 1991 and 2016. The Kaplan-Meier method and log-rank tests were used to evaluate the survival. Univariate and multivariate Cox regression analysis were used to identify prognostic biomarkers for overall survival (OS) and disease-specific survival (DSS). Furthermore, a prognostic nomogram was established, and its predictive accuracy and discriminative ability were determined using the concordance index (C-index). RESULTS: In total, 245 patients diagnosed with MC-SG were identified. The median OS was 152.0 months, with 3-, 5-, and 10-year survival rates of 79.8%, 69.2%, and 50.3%. The 3-, 5-, and 10-year DSS rates were 82.5%, 77.1%, and 61.9%, respectively. Regarding the treatment regimen, most patients (92.2%) underwent surgery, and 103 patients (42.4%) received postoperative radiotherapy. Surgery could significantly prolong OS and DSS (p < .05), but postoperative radiotherapy did not significantly prolong OS and DSS when compared with individuals receiving surgery alone (p > .05). Multivariate Cox analysis revealed that T category (T4), lymph node metastasis (N2), distant metastasis (M1), and poor differentiation were independent unfavorable prognostic factors for OS and DSS. Older age (>62 years) was also independently associated with OS. In addition, the C-index for the established OS- and DSS-specific nomogram was 0.80 (95% CI: 0.72-0.88) and 0.82 (95% CI: 0.73-0.90). CONCLUSION: Age, tumor invasion, metastases, and pathological grade were independently associated with prognosis of MC-SG patients, and the prognostic nomogram of this rare disease was established.

Survival and risk factors of adenosquamous carcinoma in the oral and maxillofacial region: a population-based study.

BACKGROUND: The features and prognosis of adenosquamous carcinoma (ASC) in oral and maxillofacial region have not thoroughly investigated, the purpose of this study is to describe clinicopathologic characteristics, treatment, and prognostic factors of this disease. METHODS: The data of 276 patients diagnosed with ASC in oral and maxillofacial region between 1975 and 2016 were collected from the Surveillance, Epidemiology, and End Results (SEER) database. The prognostic factors influencing overall survival (OS) and disease-specific survival (DSS) were identified by the Kaplan-Meier analysis and Cox regression analysis. The nomograms for OS and DSS were constructed to predict the prognosis of these patients. RESULTS: Of 276 included patients, 62.7% were male and 37.3% were female, with an average age at diagnosis of 63.5 years. The most common primary site is oral cavity (170/276), followed by salivary gland (106/276). The 3-, and 5-year OS of patients with ASC in oral and maxillofacial region were 49.0% and 38.9%, while the 3-, and 5-year DSS were 67.7%, and 60.4%, respectively. Patients who underwent surgery had longer OS (mOS: 58 m vs. 8 m) and DSS (mDSS: 193 m vs. 18 m) than those who did not. Age, AJCC-T/N/M category as well as surgery were independently associated with OS. Advanced T stage, distant metastases, and surgery were independent factors for DSS. The prognostic nomograms for OS and DSS were constructed, and the C-indexes were 0.71 (95% CI 0.66-0.76) and 0.76 (95% CI 0.67-0.85), respectively. CONCLUSION: Surgery was the favorable prognostic factor for both OS and DSS among patients with ASC in oral and maxillofacial region.

A mimetic peptide of α2,6-sialyllactose promotes neuritogenesis.

Oxidative stress contributes to the pathogenesis of neurodegenerative diseases. With the aim to find reagents that reduce oxidative stress, a phage display library was screened for peptides mimicking α2,6-sialyllactose (6'-SL), which is known to beneficially influence neural functions. Using Sambucus nigra lectin, which specifically binds to 6'-SL, we screened a phage display library and found a peptide comprising identical sequences of 12 amino acids. Mimetic peptide, reverse peptide and scrambled peptide were tested for inhibition of 6'-SL binding to the lectin. Indeed, lectin binding to 6'-SL was inhibited by the most frequently identified mimetic peptide, but not by the reverse or scrambled peptides, showing that this peptide mimics 6'-SL. Functionally, mimetic peptide, but not the reverse or scrambled peptides, increased viability and expression of neural cell adhesion molecule L1 in SK-N-SH human neuroblastoma cells, and promoted survival and neurite outgrowth of cultured mouse cerebellar granule neurons challenged by H2O2-induced oxidative stress. The combined results indicate that the 6'-SL mimetic peptide promotes neuronal survival and neuritogenesis, thus raising hopes for the treatment of neurodegenerative diseases. This study was approved by the Medical Ethics Committee of Shantou University Medical College, China (approval No. SUMC 2014-004) on February 20, 2014.

Development and in vitro study of a bi-specific magnetic resonance imaging molecular probe for hepatocellular carcinoma.

BACKGROUND: Hepatocellular carcinoma (HCC) ranks second in terms of cancer mortality worldwide. Molecular magnetic resonance imaging (MRI) targeting HCC biomarkers such as alpha-fetoprotein (AFP) or glypican-3 (GPC3) offers new strategies to enhance specificity and help early diagnosis of HCC. However, the existing iron oxide nanoparticle-based MR molecular probes singly target AFP or GPC3, which may hinder their efficiency to detect heterogeneous micro malignant HCC tumors < 1 cm (MHCC). We hypothesized that the strategy of double antibody-conjugated iron oxide nanoparticles which simultaneously target AFP and GPC3 antigens may potentially be used to overcome the tumor heterogeneity and enhance the detection rate for MRI-based MHCC diagnosis. AIM: To synthesize an AFP/GPC3 double antibody-labeled iron oxide MRI molecular probe and to assess its impact on MRI specificity and sensitivity at the cellular level. METHODS: A double antigen-targeted MRI probe for MHCC anti-AFP-USPIO-anti-GPC3 (UAG) was developed by simultaneously conjugating AFP andGPC3 antibodies to a 5 nm ultra-small superparamagnetic iron oxide nanoparticle (USPIO). At the same time, the singly labeled probes of anti-AFP-USPIO (UA) and anti-GPC3-USPIO (UG) and non-targeted USPIO (U) were also prepared for comparison. The physical characterization including morphology (transmission electron microscopy), hydrodynamic size, and zeta potential (dynamic light scattering) was conducted for each of the probes. The antigen targeting and MRI ability for these four kinds of USPIO probes were studied in the GPC3-expressing murine hepatoma cell line Hepa1-6/GPC3. First, AFP and GPC3 antigen expression in Hepa1-6/GPC3 cells was confirmed by flow cytometry and immunocytochemistry. Then, the cellular uptake of USPIO probes was investigated by Prussian blue staining assay and in vitro MRI (T2-weighted and T2-map) with a 3.0 Tesla clinical MR scanner. RESULTS: Our data showed that the double antibody-conjugated probe UAG had the best specificity in targeting Hepa1-6/GPC3 cells expressing AFP and GPC3 antigens compared with single antibody-conjugated and unconjugated USPIO probes. The iron Prussian blue staining and quantitative T2-map MRI analysis showed that, compared with UA, UG, and U, the uptake of double antigen-targeted UAG probe demonstrated a 23.3% (vs UA), 15.4% (vs UG), and 57.3% (vs U) increased Prussian stained cell percentage and a 14.93% (vs UA), 9.38% (vs UG), and 15.3% (vs U) reduction of T2 relaxation time, respectively. Such bi-specific probe might have the potential to overcome tumor heterogeneity. Meanwhile, the coupling of two antibodies did not influence the magnetic performance of USPIO, and the relatively small hydrodynamic size (59.60 ± 1.87 nm) of double antibody-conjugated USPIO probe makes it a viable candidate for use in MHCC MRI in vivo, as they are slowly phagocytosed by macrophages. CONCLUSION: The bi-specific probe presents enhanced targeting efficiency and MRI sensitivity to HCC cells than singly- or non-targeted USPIO, paving the way for in vivo translation to further evaluate its clinical potential.

Efficacy of Mobile Health Care Application and Wearable Device in Improvement of Physical Performance in Colorectal Cancer Patients Undergoing Chemotherapy.

BACKGROUND: The use of a mobile health care application, the delivery of health care or health care-related services through the use of portable devices, to manage functional loss, treatment-related toxicities, and impaired quality of life in cancer patients during chemotherapy through supervised self-management has been increasing. The aim of the present study was to evaluate the efficacy and feasibility of comprehensive mobile health care using a tailored rehabilitation program for colorectal cancer patients undergoing active chemotherapy. PATIENTS AND METHODS: A total of 102 colorectal cancer patients undergoing chemotherapy underwent 12 weeks of smartphone aftercare through provision of a mobile application and wearable device that included a rehabilitation exercise program and information on their disease and treatment. The grip strength test, 30-second chair stand test, 2-minute walk test, amount of physical activity (International Physical Activity Questionnaire short-form), quality of life (European Organization for Research and Treatment of Cancer Quality of Life Questionnaire C30), and nutritional status (Patient-generated Subjective Global Assessment) were assessed and measured at baseline, at mid-intervention (6 weeks), and at completion of the intervention (12 weeks). The rehabilitation exercise intensity was adjusted by the test results at every assessment and through real-time communication between the patients and clinicians. RESULTS: Of the 102 patients, 75 completed all 12 weeks of the smartphone aftercare rehabilitation program. The lower extremity strength (P < .001) and cardiorespiratory endurance (P < .001) was significantly improved. Fatigue (P < .007) and nausea/vomiting (P < .040) symptoms were significantly relieved after the program. CONCLUSION: A tailored rehabilitation exercise program provided through a comprehensive mobile health care application was effective in improving patients' physical capacity and treatment-related symptoms even during active chemotherapy.

Sub-chronic lead and cadmium co-induce apoptosis protein expression in liver and kidney of rats.

The combined subchronic effects of exposure to lead acetate and cadmium chloride on apoptosis protein expression were detected in the liver and kidney of rats to investigate the hazards of environmentally relevant, low-dose exposure to these compounds. The TUNEL assay showed that there were increased numbers of apoptotic cells. Immunohistochemical tests showed increased numbers of positive cells under Bax and caspase-3 protein detection and decreased Bcl-2 protein. Furthermore, mitochondrial injury and increased numbers of apoptotic cells with condensed nuclei were observed by TEM. These results suggested that low-dose exposure to Pb and Cd can cause significant hepatic and renal apoptosis and finally impair their function. Hepatic and renal apoptosis induced by low-dose exposure is associated with mitochondrial injury and changes in levels of apoptogenic proteins, such as Bcl-2, Bax, and caspase-3.

Cells with dysfunctional telomeres are susceptible to reactive oxygen species hydrogen peroxide via generation of multichromosomal fusions and chromosomal fragments bearing telomeres.

During genotoxic stress, reactive oxygen species hydrogen peroxide (H(2)O(2)) is a prime mediator of the DNA damage response. Telomeres function both to assist in DNA damage repair and to inhibit chromosomal end-to-end fusion. Here, we show that telomere dysfunction renders cells susceptible to H(2)O(2), via generation of multichromosomal fusion and chromosomal fragments. H(2)O(2) caused formation of multichromosomal end-to-end fusions involving more than three chromosomes, preferentially when telomeres were erosive. Interestingly, extensive chromosomal fragmentation (yielding small-sized fragments) occurred only in cells exhibiting such multichromosomal fusions. Telomeres were absent from fusion points, being rather present in the small fragments, indicating that H(2)O(2) cleaves chromosomal regions adjacent to telomeres. Restoration of telomere function or addition of the antioxidant N-acetylcysteine prevented development of chromosomal aberrations and rescued the observed hypersensitivity to H(2)O(2). Thus, chromosomal regions adjacent to telomeres become sensitive to reactive oxygen species hydrogen peroxide when telomeres are dysfunctional, and are cleaved to produce multichromosomal fusions and small chromosomal fragments bearing the telomeres.

Involvement of calcitonin gene-related peptide innervation in the promoting effect of low-intensity pulsed ultrasound on spinal fusion without decortication.

STUDY DESIGN: A prospective left-right comparison designed experiment using a rabbit posterolateral intertransverse process fusion model. OBJECTIVE: To investigate the involvement of calcitonin gene-related peptide (CGRP) innervation in the promoting effect of low intensity pulsed ultrasound stimulation (LIPUS) on spinal fusion without decortication. SUMMARY OF BACKGROUND DATA: Sensory neuropeptide CGRP is involved in bone repair and ectopic ossification. Comparison of CGRP innervations in ectopic bone between sham LIPUS and LIPUS sides can help us to understand the relationship between sensory nerve innervation and LIPUS. METHODS: A total of 27 New Zealand white rabbits underwent bilateral posterolateral intertransverse process fusion with implantation of porous poly-D,L-lactic acid blocks loaded with 1.25 µg recombinant human bone morphogenetic protein-4 solution. One side was provided LIPUS daily whereas the other side served as control. Animals were killed and the operated lumbar vertebrae were harvested for histomorphologic evaluation at 3 days (n = 3), 1 week (n = 6), 3 weeks (n = 6), 7 weeks (n = 6), and 12 weeks (n = 6) following surgery, respectively. RESULTS: LIPUS accelerated the invasion of CGRP-positive nerve fibers during ectopic ossification spatially and temporally. Spatially, CGRP-positive nerve fibers were also observed in the new formed cartilage and bone tissues on LIPUS side, whereas they were only detected in the fibrous tissue and bone marrow on sham LIPUS side. Temporally, the density of CGRP-positive nerve fibers was significantly higher on the LIPUS side when compared with the sham LIPUS side. CONCLUSION: LIPUS promoted the invasion of CGRP sensory nerve in ectopic bone, which may in turn contribute to the promoting effect of LIPUS on ectopic ossification.

Serum obestatin/ghrelin ratio is altered in patients after distal gastrectomy.

BACKGROUND/AIMS: Ghrelin is a peptide hormone produced mainly in the stomach, and obestatin is derived by proteolytic cleavage of the ghrelin prepro-hormone. The aim of this study was to determine the postoperative serial changes in these hormones and whether hyperplasia of ghrelin-expressing cells occurs in the remnant stomach. METHODS: We prospectively analyzed serial serum samples of 45 early gastric cancer patients and remnant stomach samples of 24 patients. RESULT: The serum obestatin level on day 2 was lower than that on day 0, and it subsequently returned to the level observed on day 0. In contrast, the serum ghrelin level was lower on days 120 and 210 than on day 0. Eventually, the obestatin/ghrelin ratio was significantly high on day 210 (p = 0.0003). Moreover, we did not observe an increase in the number of ghrelin-expressing cells. The number of ghrelin-expressing cells correlated with the serum ghrelin level. CONCLUSION: The serum level of obestatin and ghrelin exhibits a different time course in patients who have undergone gastrectomy, and there was no ghrelin-expressing cell hyperplasia in the remnant stomach despite the decrease in serum ghrelin.

Mutation analysis of p31comet gene, a negative regulator of Mad2, in human hepatocellular carcinoma.

Failure of mitotic checkpoint machinery leads to the chromosomal missegregation and nuclear endoreduplication, thereby driving the emergence of aneuploidy and tetraploidy population. Although abnormal nuclear ploidy and the resulting impairment of mitotic checkpoint function are typical physiological event leading to human hepatocellular carcinoma, any mutational change of mitotic checkpoint regulators has not yet been discovered. Therefore, we investigated the mutation of p31(comet), a recently identified mitotic checkpoint regulator, in human hepatocellular carcinoma. Of 51 human hepatocellular carcinoma tissue and 6 cell lines tested, five samples exhibited nucleotide sequence variations dispersed on four sites within the entire coding sequence. Among these sites with sequence substitutions, three were found to be missense mutation accompanied with amino acid change but one was a silent mutation. Of these sequence substitutions, two were present in both tumor and non-tumor liver tissues, suggesting the possibility of polymorphism. The present findings indicate that p31(comet) does not have an impact on the formation of aneuploidy and tetraploidy found in human hepatocellular carcinoma.

[The prevalence and clinical characteristics of hepatitis-delta infection in Korea].

BACKGROUND/AIMS: The prevalence of hepatitis delta virus (HDV) infection has been estimated as being approximately 5% among global HBsAg carriers. The anti-delta positive rate in Koreans had been reported as being 0.85% in 1985. While the prevalence of HBV has been decreased from nearly 10% to 5% during the past twenty years, there have been no more studies on the anti-delta prevalence in Koreans. The aim of this study was to estimate the anti-delta prevalence in Koreans and to study the clinical characteristics of anti-delta positive patients in a single center. METHODS: Serum anti-delta was measured in one hundred ninety four HBsAg-positive patients who were admitted to our hospital from February 2003 to August 2003. We checked the genotypes of the HBV in the anti-delta positive patients. The clinical features of the anti-delta positive patients were compared to those clinical features of the anti-delta negative patients from the aspect of age, gender, mode of transmission, the positivity of HBeAg and serum HBV DNA. RESULTS: Serum anti-delta was positive in seven patients among the 194 subjects, giving a 3.6% positive rate. Among these seven patients, six had hepatocellular carcinoma (HCC) and the other one had cholangiocarcinoma. All of the anti-delta positive patients had the C genotype of HBV. The anti-delta positive patients showed significantly suppressed HBV DNA replication compared to the anti-delta negative patients. CONCLUSIONS: In Koreans, anti-delta was positive mainly in HCC patients with an approximate prevalence of 4%, and this rate has not changed much for the past twenty years. HBV DNA replication was suppressed by HDV infection.