The study on specific umbilical blood Dc vaccine for Beige nude mice loaded human colorectal carcinoma to induce anti-tumor immunity.

OBJECTIVE: This study is to observe the immunosuppression of CD137L transfected umbilical blood Dcs (Dendritic cell) vaccine to tumor development of SCID/ Beige nude mice. MATERIALS AND METHODS: Samples of umbilical blood in the childbirth pregnant women were collected by density gradient centrifugation. Umbilical cord blood dendritic cells (Dcs) were transfected by specific CD137L via LipofectamineTM method and cells were harvested. Meanwhile, the peripheral blood of volunteers was collected to isolate Dcs, the Dcs were cultured for 5 days and hatched with SW-1116 cells antigen. The mature Dcs were harvested. The male SCID/Beige nude mice were subcutaneously injected with human SW-1116 cells in axillary to build colorectal carcinoma model as blank control (Blank). The naked peripheral blood Dc vaccine group (cPBMCs), the SW-1116 antigen-specific peripheral blood Dc vaccine group (pDcs) and the CD137L specific umbilical blood Dc vaccine group (tuDcs) were injected 24 h before tumor cells injection, respectively to recur the humanized immune reconstruction. The general life, living habits changes, tumor growing time and tumor size were observed. The nude mice were sacrificed 18 days after tumor formation. The tumor size, mice weight, in vitro tumor weight, liver weight and spleen weight of mice were recorded to evaluate the anti-tumor effect of the specific immune cells. RESULTS: The nude mice in pDcs group showed better general living condition, slower tumor growth, smaller tumor volume and no ulceration, necrosis, and death in nude mice. The tumor formation time in different groups was 4.71 ± 0.18 ds (blank), 7.71 ± 0.29 ds (cPBMCs), 7.86 ± 0.26 ds (pDcs) and 8.14 ± 0.69 ds (tuDcs) respectively. There were significant differences between blank and other three groups (F = 40.96, p < 0.01). Compared to mice in blank group, the tumor volume of cPBMCs group was significantly smaller (201.43 ± 69.84 mm³ vs. 436.04 ± 54.50 mm³, p < 0.01) and the tumor weight were significantly smaller (1.25 ± 0.12 g vs. 2.83 ± 0.24 g, p < 0.01). The tumor volume of tuDcs mice was significantly smaller than that of blank (92.11 ± 11.55 mm³ vs. 436.04 ± 54.50 mm³, p < 0.01) and cPBMCs mice (92.11 ± 11.55 mm³ vs. 201.43 ± 69.84 mm³, p < 0.01). Similarly, the tumor weight of tuDcs mice was significantly smaller than that of blank (0.66 ± 0.07 g vs. 2.83 ± 0.24 g, p < 0.01) and cPBMCs mice (0.66 ± 0.07 g vs. 1.25 ± 0.12 g, p < 0.01). There was no significant difference in tumor volume (92.11 ± 11.55 mm³ vs. 85.61 ± 11.59 mm³, p = 0.69) and tumor weight (0.66 ± 0.07 g vs. 0.63 ± 0.09 g, p = 0.75) between tuDcs group and pDcs group. CONCLUSIONS: The specific CD137L transfected umbilical blood Dc vaccine had significant anti-tumor effect against human colon cancer in nude mice via increasing the number of immune effector cell in tumor microenvironment.

Hippocampal overexpression of Down syndrome cell adhesion molecule in amyloid precursor protein transgenic mice.

Down syndrome cell adhesion molecule (DSCAM) is located within the Down syndrome critical region of chromosome 21. DSCAM is a broadly expressed neurodevelopmental protein involved in synaptogenesis, neurite outgrowth, and axon guidance. We previously demonstrated DSCAM overexpression in the cortex of amyloid precursor protein (APP) transgenic mice, suggesting possible regulatory interactions between APP and DSCAM. APP mice exhibit deficits in hippocampus-dependent learning and memory. In this preliminary study, we examined age-related changes in DSCAM expression within the hippocampus in 16 APP transgenic mice (1, 3, 6 and 12 months old). Hippocampus-dependent spatial memory was assessed in APP mice and age-matched wild type littermates (WTs) using the Morris water maze (MWM). The cellular distribution of hippocampal DSCAM and total expression at both mRNA and protein levels were measured by immunohistochemistry, qRT-PCR, and western blotting, respectively. APP mice exhibited spatial memory deficits in the MWM. Intense DSCAM immunoreactivity was observed in the dentate gyrus granule cell layer and hippocampal stratum pyramidale. Total hippocampal DSCAM mRNA and protein expression levels were substantially higher in APP mice than WTs at 1 and 3 months of age. Expression decreased with age in both groups but remained higher in APP mice. DSCAM is overexpressed in the hippocampus over the first 12 months of life in APP mice, but especially during maturation to adulthood. In conclusion, these results suggest an association between DSCAM and APP mice, which is characterized by neuropathology and behavioral deficits. These results provide some clues for future studies on the role of DSCAM overexpression in the precocious cognitive decline observed in APP transgenic mice.

[Changes on patterns of sleep duration: findings from China Health and Nutrition Survey in population in 9 provinces].

Objective: To understand the changes on patterns of sleep duration of the China Health and Nutrition Survey (CHNS) cohort in 9 provinces from 2004 to 2011. Methods: Four rounds of CHNS data were used. Urban/rural, age and gender specific insufficient sleeping rates and excessive sleeping rates were analyzed. Results: In 2004, 2006, 2009 and 2011, a total of 274, 281, 329 and 304 children aged 3-5 years; 874, 806, 768 and 742 children aged 6-12 years; 789, 529, 426 and 367 children aged 13-17 years; 9 568, 9 530, 9 942 and 9 609 adults aged ≥18 years were surveyed respectively. The lowest insufficient sleeping rate was 53.9% (200/371) in 3-17 years old children in rural area in 2006, the highest insufficient sleeping rate was 77.2% (44/57) in 3-5 years old children in urban area in 2004. The insufficient sleeping rate increased in rural 3-5 years old children from 2004 to 2011. For the adults aged ≥18 years, the insufficient sleeping rate ranged from 4.2% (82/1 954) in females aged 18-44 years in 2004 and 2009 to 20.8% (211/1 015) in urban residents aged > 60 years in 2011. The insufficient sleeping rate in age-groups 44-59 years and ≥60 years increased in both males and females and in both urban area and rural area from 2004 to 2011. The gender specific excessive sleeping rate in 3-17 years old children was very low in both urban area and rural area and no difference was found in different rounds of survey. The excessive sleeping rate in adults ranged from 18.4% (569/3 093) in urban population in 2011 to 32.5% (1 617/4 969) in females in 2004. The excessive sleeping rate of adult decreased from 2004 to 2011. Conclusion: We should pay attention to the fact that the insufficient sleeping rate in adolescents is high and in increase in rural 3-5 years old children and adults aged ≥45 years.

Peroxiredoxins in colorectal neoplasms.

Peroxiredoxins (Prxs) are novel group proteins with efficient antioxidant capacity, and some of them also have effects on cell proliferation, differentiation, apoptosis, and chemotherapy and radiotherapy resistance. Altogether six distinct Prxs expressions were investigated in histological samples of colorectal neoplasm and the distant normal tissues and investigated associatedly with parameters such as clinical stage and lymphnodes metastasis. Normal colorectal tissues were almost negative for Prxs, except Prx4 (15/32). In colorectal cancer tissues, the most prominent reactivity was observed with Prx2 in 23/32 cases, while the corresponding figures for others was 21/32 (Prx1), 18/32 (Prx3, Prx5, Prx6) and 8/32 (Prx4). Prx1 (P=0.023), Prx2 (P=0.012), and Prx5 (P=0.028) were the isoforms that showed significantly increased expression in colorectal cancer patients with stage III or lymphnodes metastasis-positive cases. There was a significant relationship between the expression of Prx1 and Prx2 (rs=0.425, P=0.015) and between Prx3 and Prx4 (rs=0.364, P=0.041). Additionally, 8 cases were studied by western analysis. Prx1, 2, 3, 5 and 6 were particularly elevated in tumors compared to nonmalignant tissue as assessed by immunohistochemistry. It appeared that some Prxs were upexpression in colorectal cancer tissues and may have some prognostic significance; the induction of Prxs could be explained by increased production of reactive oxygen species in carcinomatous tissue.

Identification of differential proteins in colon cancer SW480 cells with HIF1-alpha silence by proteome analysis.

Hypoxia is one of the basic characteristics of the colorectal cancer and HIF1 plays a central role in tumor hypoxia adaptation and controls the expression of a variety of genes. But it had not reached a comprehensive understanding of them in colorectal cancer in hypoxia. This study silenced HIF1-alpha, which is the regulatory subunit of HIF1, and established two dimensional gel electrophoresis profiles for human SW480siHIF1-alpha and SW480N cells to identified proteins with differential expressions. Ten down-regulated proteins and nine up-regulated proteins were identified by HPLC-Chip-MS/MS. Down-regulated proteins including PLD2, ANXA2. HIF1-alpha had a clear positive correlation with PLD2 and ANXA2 in colon adenocarcinoma samples. There proteins might have a direct or indirect contact HIF1-alpha, and play a critical role in apoptosis, signal transduction and hypoxia adaptation.

Synthesis of GaN nanotip triangle pyramids on 3C-SiC epilayer/Si substrates via an in situ In-doping technique.

GaN nanotip triangle pyramids were synthesized on 3C-SiC epilayer via an isoelectronic In-doping technique. The synthesis was carried out in a specially designed two-hot-boat chemical vapor deposition system. In (99.999%) and molten Ga (99.99%) with a mass ratio of about 1:4 were used as the source, and pieces of Si (111) wafer covered with 400-500 nm 3C-SiC epilayer were used as the substrates. The products were analyzed by x-ray diffraction, field emission scanning electron microscopy, high-resolution transmission electron microscopy, selected area electron diffraction, Raman spectroscopy, and photoluminescence measurements. Our results show that the as-synthesized GaN pyramids are perfect single crystal with wurtzite structure, which may have potential applications in electronic/photonic devices.

Extension of ischemic tolerance of porcine livers by cold preservation including postconditioning with gaseous oxygen.

BACKGROUND: Transplantation of organs from nonheartbeating donors was recommended to reduce organ shortage. In vitro experiments with rat livers have shown that the warm ischemic tolerance of the liver may be extended by persufflation with gaseous oxygen during cold storage. The qualification of this method for procurement of livers harvested after cardiac arrest was tested in an in vivo approach with pigs. METHODS: Livers from 15 donor pigs were explanted, heparinized, flushed with and stored in University of Wisconsin solution for 4 hr at 4 degrees C, and then implanted into 15 recipients. The organs were dissected immediately after cardiac arrest (group 1) or after 60 min of warm ischemia (groups 2 and 3). Group 2 livers received 75,000 IU of superoxide dismutase together with the flush solution and were persufflated with gaseous oxygen via the venous vascular system during cold storage. Main end point was survival after 5 days. Additionally, metabolic, functional and inflammatory criteria were measured in the blood. RESULTS: All animals of the groups 1 and 2 survived, all animals of group 3 died within 3 hr after reperfusion. In all groups the blood parameters reflected significant damage of the livers. However, the ischemic damage was comparable in the groups 1 and 2 whereas the livers of group 3 exhibited significantly higher levels of aspartate alanine aminotransferase and lactate dehydrogenase, and a significantly elongated partial thromboplastin time 1 hr after reperfusion (P=0.016). CONCLUSIONS: Venous systemic oxygen persufflation in combination with antioxidative medication is a promising new method of resuscitating ischemically altered livers from nonheartbeating donors for successful transplantation.

Evaluation of the effects of a pneumoperitoneum with carbon dioxide or helium in a porcine model of endotoxemia.

BACKGROUND: The expansion of the laparoscopic techniques to the critically ill patient is currently under debate. In order to evaluate the potential risks of performing laparoscopy in a body with signs of sepsis, the effects of the pneumoperitoneum were studied in a porcine model of mild endotoxemia. METHODS: Twenty-eight pigs were separated into four groups of seven animals: untreated control (C), 2 microg/kg/h endotoxin (E), endotoxin and a pneumoperitoneum (3 h, 15 mmHg) with CO2 (EC) or with helium (EH). Hemodynamic and homeostatic variables were studied for a period of 5.5 h. Primary endpoints were arterial and mucosal pH and the ATP content of the bowel wall. Statistical evaluation was performed using analysis of variance and the Bonferroni test. RESULTS: Endotoxin infusion induced characteristic symptoms of early sepsis: increase of arterial CO2, pulmonary arterial, portal venous, and pulmonary artery wedge pressure, and decrease of arterial pressure, cardiac output, arterial and mucosal pH. An additional pneumoperitoneum led to aggravation of all criteria with significant alterations in arterial and mucosal pH, arterial CO2, wedge and portal venous pressure. The most striking derangement of mean values was observed for mucosal pH (EC: 7.40, EH: 7.54) and arterial pH (EC: 7.15, EH: 7.18). In group EC, two animals died in septic shock. CONCLUSION: Applying a pneumoperitoneum during an ongoing sepsis significantly deteriorates hemodynamic and homeostatic variables, thus enhancing the risk of severe complications.

Preoperative high dose methylprednisolone improves patients outcome after abdominal surgery.

OBJECTIVE: To assess the effect of preoperative high dose methylprednisolone on stress response and outcome. DESIGN: Randomised, placebo-controlled, double-blind study. SETTING: University hospital, Germany. SUBJECTS: 20 patients listed for abdominal surgery of whom 10 had major intra-abdominal interventions and 10 had incisional hernias repaired. INTERVENTIONS: Methylprednisolone 30 mg/kg (100 ml) was given by slow intravenous infusion 90-60 minutes before operation. The control group received the same volume of sodium chloride. MAIN OUTCOME MEASURES: Speed of convalescence, degree of fatigue, amount of pain, consumption of analgesics, breathing capacity, and hospital stay, as well as humoral and cellular mediators of the stress response. RESULTS: Methylprednisolone significantly improved criteria of postoperative recovery, fatigue by 47%, (day 1), convalescence by about 45% (days 1-3), and breathing capacity (FEV1) between 47% and 29% (days 5, 7) (p < 0.05, ANOVA), and led to a significant reduction of median hospital stay of 4.5 days. C-reactive protein concentration was significantly decreased (by 46% on day 3) and T-cell activation was suppressed (day 1). CONCLUSION: Outcome of the patients after conventional abdominal surgery is substantially improved by preoperative high dose methylprednisolone. This effect is more pronounced in patients having major operations.

[Effect of pneumoperitoneum on hemodynamics and intestinal integrity in endotoxinemia: CO2 versus helium]. / Einfluss eines Pneumoperitoneums auf Hämodynamik und intestinale Integrität bei Endotoxinämie: CO2 versus Helium.

In a study in pigs pathophysiological effects of a pneumoperitoneum (PP) established during endotoxemia (2 micrograms/kg b.w./hr) were investigated. In this situation a PP with carbon dioxide (CO2) or helium induces a deterioration of systemic hemodynamics or acid-base-stability. Some of these effects threaten vital stability. PP with either gas causes a mucosal acidosis of the small bowel within the meaning of an impairment of intestinal oxygen supply. A CO2-PP adversely affects the intestinal energy metabolism and leads to significantly increased endotoxin levels in contrast to helium after desufflation of the abdominal cavity. Data of this study at least demand for a careful monitoring, if laparoscopy is performed in septic and critically ill patients.

[Revitalizing donor livers after cardiovascular arrest with venous oxygen persufflation]. / Revitalisierung von Spenderlebern nach Kreislaufstillstand mittels venöser Sauerstoffpersufflation.

The experimental study investigates the resuscitation of livers procured from non heart beating donors by venous systemic oxygen persufflation in a pig transplantation model. In group 1 livers were harvested from donor animals after 60 min of potassium induced cardiac arrest. Livers were then flushed and preserved in 4 degrees C cold University of Wisconsin solution for 4 hrs. Then liver transplantation was performed in recipient animals. Animals of group 2 received the same treatment, but during hypothermic storage we applied venous systemic oxygen gas persufflation (VSOP) via the supra-hepatic vena cava. Livers of group 3 were harvested without warm ischemia and without oxygen treatment during cold ischemic storage and were then transplanted. All recipient animals of group 2 and 3 survived the five day follow-up time. All recipient animals of group 1 died a few hours after liver reperfusion due to circulatory shock or liver failure. The liver quality of group 2 recipient animals was similar to group 3 livers, because serum transaminase levels and blood coagulation test showed no significantly different values between those groups. Our data suggest that livers procured from non heart beating donors after 1 hr of cardiac arrest can be resuscitated and successfully transplanted by therapeutic treatment with VSOP.

Antibodies against Tetrahymena 14-nm filament-forming protein recognize the replication band in Euplotes.

Tetrahymena 14-nm filament-forming protein (49K protein) is a structural protein which is involved in activity of the pronuclei during conjugation (O. Numata, T. Sugai, and Y. Watanabe (1985) Nature (London) 314, 192-194). Using monoclonal and polyclonal antibodies, we here demonstrate the presence of a cross-reactive protein (CRP-49) within the macronuclear replication bands of Euplotes harpa and E. eurystomus which is recognized by anti-49K protein antibodies. Immunoblotting reveals that both monoclonal and polyclonal antibodies cross-react to a protein with an apparent molecular mass of 50 kDa in an E. harpa cell extract and to a protein of 49 kDa in a macronuclear extract of E. eurystomus. The antibodies used in this study have no effect upon in vitro DNA synthesis in the replication band of E. eurystomus.