OBJECTIVEMeasurement of biomarkers is a potential approach to early prediction of the risk of mortality in patients with sepsis. The aim of the present study was to evaluate the prognostic value of pro-atrial natriuretic peptide (pro-ANP) and pro-adrenomedullin (pro-ADM) levels in a cohort of medical intensive care patients and to compare it with that of other known biomarkers and physiological scores.

METHODSBlood samples of 51 consecutive critically ill patients admitted to the intensive care unit and 53 age-matched healthy control people were evaluated in this prospective study. The prognostic value of pro-ANP and pro-ADM levels was compared with that of acute physiology and chronic health evaluation (APACHE) II scores and various biomarkers such as C-reactive protein, interleukin-6 and procalcitonin. Pro-ANP and pro-ADM were detected by a new sandwich immunoassay.

RESULTSOn admission, 25 patients had systemic inflammatory response syndrome (SIRS), 12 sepsis, 9 severe sepsis and 5 septic shock. At that time, the median levels (ng/ml) of pro-ANP and pro-ADM were 87.22 and 0.34 respectively in patients with SIRS, 1533.30 and 2.23 in those with sepsis, 1098.73 and 4.57 in those with severe sepsis, and 1933.94 and 8.21 in those with septic shock. With the increasing severity of disease, the levels of pro-ANP and pro-ADM were gradually increased. On admission, the circulating levels of pro-ANP and pro-ADM in patients with sepsis, severe sepsis, or septic shock were significantly higher in non-survivors than in survivors (P less than 0.05). In a receiver operating characteristic curve analysis for the survival of patients with sepsis, the areas under the curve (AUCs) for pro-ANP and pro-ADM were 0.89 and 0.87 respectively, which was similar to the AUCs for procalcitonin and APACHE II scores.

CONCLUSIONPro-ANP and pro-ADM are valuable biomarkers for prediction of severity of septic patients.

APACHE ; Adolescent ; Adrenomedullin ; blood ; Adult ; Aged ; Atrial Natriuretic Factor ; blood ; C-Reactive Protein ; analysis ; Female ; Humans ; Male ; Middle Aged ; Protein Precursors ; blood ; Sepsis ; blood ; Shock, Septic ; blood

To review the challenges and countermeasures in the hospital care for Wenchuan earthquake casualties and draw lessons for the protective response in the future. Medical records and laboratory findings of the victims admitted in West China Hospital (WCH) were retrospectively analyzed. Related data were compared between beforemath and aftermath of the earthquake and between WCH and frontier county hospitals. One thousand and thirty-one earthquake survivors were hospitalized, 1 358 victims underwent surgery and 142 victims were transferred to intensive care unit. The incidence of infection, crush syndrome and multiple organ dysfunction syndrome (MODS) was 39.6%, 20.7% and 2.3% respectively. Wound classification showed that the incidence of extremity damage was 72%, while the incidence of chest trauma, abdominal trauma and brain trauma was less than 10% respectively. Isolating rates of environmental pathogens were increased in the aftermath of earthquake, and the spectrum of the pathogens and related antibiotic sensitivities were quite different from those in the beforemath of earthquake. The social economic and population conditions in the earthquake-stricken areas affected the composition of the victims and the geographic features restricted the efficiency of rescue. Trauma-induced MODS, crush syndrome and severe infections all constituted the dilemma for the hospital care, to resolve whether the multidiscipline team work was proved to be an optimizing choice. For a more effective disaster protective response in the future, the study on rescue plan and the ladder therapies for massive casualties should be potentiated.
Adolescent ; Adult ; Aged ; Bacterial Infections ; epidemiology ; Child ; Child, Preschool ; China ; epidemiology ; Communicable Disease Control ; Crush Syndrome ; epidemiology ; Earthquakes ; Female ; Hospitals ; Humans ; Infant ; Infant, Newborn ; Male ; Middle Aged ; Multiple Organ Failure ; epidemiology ; Rescue Work ; Young Adult

Animals ; Bone and Bones ; cytology ; metabolism ; pathology ; Collagen ; metabolism ; Durapatite ; metabolism ; Female ; Osteoporosis ; metabolism ; pathology ; Ovariectomy ; adverse effects ; Rats ; Rats, Sprague-Dawley

BACKGROUNDAntigen-loaded eosinophils (EOSs) instilled intratracheally into mice were capable of inducing Th2-type cytokine production in the draining lymph nodes. The aim of the present study was to evaluate whether EOSs within the tracheobronchial lumen can stimulate Th2 cell expansion in the lung tissues.

METHODSAirway EOSs were recovered from ovalbumin-sensitized and -challenged BALB/c mice, these EOSs were then cocultured with CD4+ cells isolated from sensitized mice in the absence or presence of anti-CD80 or/and -CD86 monoclonal antibodies. Airway EOSs were instilled into the trachea of sensitized mice. At the day 3 thereafter, the lung tissues were removed and prepared into cell suspensions for culture. Cell-free culture supernatants were collected for detection of cytokines.

RESULTSAirway EOSs functioned as CD80- and CD86-dependent antigen-presenting cells to stimulate lung CD4+ lymphocytes to produce interleukin-4, interleukin-5 and interleukin-13, but not interferon-gamma in in vitro assay. When instilled intratracheally in sensitized recipient mice, airway EOSs primed lung Th2 cells in vivo for interleukin-4, interleukin-5 and interleukin-13, but not interferon-gamma, production during the in vitro culture that was also CD80- and CD86-dependent.

CONCLUSIONEOSs within the lumina of airways could process inhaled antigen and function in vitro and in vivo as antigen-presenting cells to promote expansion of Th2 cells in the lungs.

Animals ; Antigen Presentation ; Antigens, CD ; physiology ; B7-1 Antigen ; physiology ; B7-2 Antigen ; Cytokines ; biosynthesis ; Eosinophils ; physiology ; Female ; Lung ; immunology ; Membrane Glycoproteins ; physiology ; Mice ; Mice, Inbred BALB C ; Th1 Cells ; immunology ; Th2 Cells ; immunology

The aim is to observe the expression of human factor VIII gene in mice tranduced in vivo and ex vivo. The vector pLNC-FVIII BD was generated by cloning a B-domain-deleted (760aa-1639aa) FVIII cDNA (FVIIIBD cDNA) into retroviral vector pLNCX. 2 x 10(6) of mouse bone marrow stroma cells transduced by LNC-FVIII BD were infused into 4-week-old BALB/c mice by tail-vein injection. pLNC-FVIII BD was conjugated with PAMAM dendrimer to form complex PAMAM-pLNC-FVIII BD, with which C57BL/6J were injected by tail vein (200 micro l contained 15 micro g/mouse) and sacrificed at days 1, 2, 7, 14, 21 and 28, respectively after injection. Tissue such as liver, spleen, lung and kindney were harvested, with which the transcription were detected by means of RT-PCR. In addition, blood was collected to be measured human FVIII Ag, human FVIIIc and anti-FVIII of human inhibitors. The results showed that the highest level of human FVIII in the recipient BALB/c mice was 8.6 +/- 1.44 ng/ml detected on the first day post-injection; anti-FVIII antibodies were detected from the first week post-injection, and then the level of FVIII Ag decreased and cannot be measured on the fourth week. In the C57BL/6J mice physiological level of human FVIII was expressed in plasma at 48 hours after injection and the average human FVIIIc was 0.62 U/ml and the average human FVIII Ag was 115.5 ng/ml, and gradually reduced later. Anti-FVIII of human inhibitors was not revealed all the time. Syngene image scanning demonstrated that the transcription of the human FVIII BD cDNA occurred mainly in spleen and lung, and secondarily in liver and kidney. No side effects of PAMAM-pLNC-FVIII BD were observed in mice tissue by pathological examination at 4 weeks. In conclusion, retrovirus-transduced bone marrow stroma cells effectively produced human FVIII after ex vivo transduction, but the development of anti-FVIII antibodies in recipient mice influenced the expression level. The human FVIII gene can successfully be transduced in vivo through injecting PAMAM-pLNC-FVIII BD cDNA into mice intravenously. There was physiological level expression of human FVIII in plasma at 48 hours after injection and the average human FVIIIc is 0.62 U/ml and the peak in the six mice was 0.89 U/ml, and gradually reduced later.
Animals ; DNA, Complementary ; analysis ; Factor VIII ; genetics ; Genetic Therapy ; Hemophilia A ; therapy ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Transfection

OBJECTIVETo construct anti-CD20scFv/CD80/CD28/zeta recombinant gene modified T cells, test its effectiveness of eradicating CD20+ lymphoma cells and provide a probably new approach to tumor adoptive immunotherapy.

METHODSCD28-zeta cDNA were amplified from vector pBULLET and inserted into pLNCX vector that contained anti-CD20scFv/CD80 gene. The recombinant vectors were transduced into PA317 cells and high titer retroviruses were obtained to infect human peripheral blood T lymphocytes. Resistant T cells were obtained by G418 selection at one week. Then transduced T lymphocytes and lymphoma cell lines Daudi Raji were cocultured. The cytotoxicity and cytokine production of transduced T cells were determined by non-radio-activation cytotoxicity assay and ELISA respectively.

RESULTSThe recombinant eukaryotic vector was constructed successfully as proved by enzyme digestion analysis and sequencing. These T cells were able to lyse CD20+ target cells and secrete high levels of IL-2 and IFN-gamma in vitro.

CONCLUSIONRecombinant gene modified T cells can be constructed successfully. It can specially kill CD20 positive lymphoma cells in vitro.

Antigens, CD20 ; genetics ; immunology ; B7-1 Antigen ; genetics ; immunology ; CD28 Antigens ; genetics ; immunology ; Cell Line ; Cytotoxicity, Immunologic ; Genetic Vectors ; Humans ; Immunotherapy, Adoptive ; Plasmids ; genetics ; T-Lymphocytes ; immunology ; metabolism ; Transfection

Objective:The aim of this study was to study the characteristics of neoplasm invasion type in ovary of two orthotopic models established with human epithelial ovarian cancer solid tumor tissue slices and human ovarian carcinoma cell line OVCAR-3 in nude mice and human epithelial ovarian cancer.Methods:Tumor tissues and cell line OVCAR-3 of human epithelial ovarian cancer were grown in subcutaneous tissue and the subcutaneous tumor source was fetched and inoculated in ovarian capsule of nude mice to establish the orthotopic implantation model. The neoplasm invasion type in the two kinds of models were observed. The neoplasm invasion types were also analyzed by pathological examination in 54 cases of International Federation of Gynecology and Obstetrics (FIGO) stageⅠ-Ⅱepithelial ovarian cancer.Results:Three neoplasm invasion types were found as follows: type of pseudocapsule, type of pseudocapsule invasion, type of pseudocapsule penetration. Pseudocapsule rate in the solid tumor slices group (18.2%) were lower than those in the cell line group (42.3%) ( P<0.05), while the pseudocapsule penetration rate in the solid tumor slices (50.0%) were higher than those in the cell line group (23.1%) ( P<0.05). No difference was found of pseudocapsule invasion rate between two groups ( P>0.05). Neoplasm invasion type in ovary changed with tumor planting time. High proportion of pseudocapsule type was found at the beginning of tumor planting, and the pseudocapsule penetration rate raised with tumor planting time increased. High proportion of pseudocapsule type was also found in patients with FIGO stageⅠepithelial ovarian cancer, and pseudocapsule penetration rate increased in those with FIGO stageⅡ. No difference in neoplasm invasion type was found between two kinds of pathological types ( P>0.05). Conclusions:There are differences between the two kinds of orthotopic models established with human epithelial ovarian cancer solid tumor tissue slices and human ovarian carcinoma cell line OVCAR-3. Compared to the solid tumor slices model, the cell line model is more stable for the follow-up study. The proportion of three neoplasm invasion types in ovary were more balanced in 8 weeks after tumor planting, and 8 weeks after tumor planting is the best start time for the follow-up experiment.

The study was aimed to compare the safety of hematopoietic stem cell mobilization and collection in related donors providing bone marrow and peripheral blood stem cells and in unrelated donors providing peripheral blood stem cells only. 100 related donors from September 2005 to August 2006 at Institute of Hematology & People Hospital, Peking University, and 71 unrelated donors from November 2003 to December 2007 in Data Bank of Chinese Hematopoietic Stem Cell Donor Beijing Management Center, were observed in process of bone marrow and peripheral blood stem cell mobilization, collection, and follow-up. The change of hematologic parameters (white blood cell count, platelet count and hemoglobin level) and the side effects were recorded and evaluated on months 1, 3 and 6 as well as annually after PBSC donation. During follow-up, long-term side effects and life quality were investigated by questionnaires. The results showed that the total MNC count of bone marrow and PBSC from related donors was 6.70 (4.11 - 12.23) x 10⁸/kg, and the total CD34(+) cell count was 3.40 (1.61 - 13.57) x 10⁶/kg; the total MNC count of PBSC from unrelated donors was 6.69 (3.35 - 11.48) x 10⁸/kg, and the total CD34(+) cell count was 3.50 (1.15 - 11.60) x 10⁶/kg. The main side effect of mobilization was bone pain, reported in 47.0% of the related donors and in 43.7% of unrelated ones, the main side effect of collection was paresthesia, reported in 25.0% of the related donors and in 29.6% of unrelated ones, there was no significant difference on side effects between related and unrelated donors during mobilization and collection of hematopoietic stem cells, all donors could endure these side effects, and no donor discontinued G-CSF administration because of side effects. After collection, the hemoglobin level of related donors was lower than that of unrelated donors [(125.8 ± 20.2) g/L vs (143.2 ± 20.1) g/L] (p < 0.05) because of bone marrow and peripheral blood collection, and the platelet count of unrelated donors were lower than that of related donors [(126.2 ± 57.2) x 10⁹/L vs (162.4 ± 72.9) x 10⁹/L] (p < 0.05) because of more than two times of collection. There was no significant difference on hematologic parameters between two groups during long-term follow-up, and the majority of the donors reported were in good or very good health. It is concluded that the donation proved from related and unrelated donors is safe to mobilize hematopoietic stem cells for allogeneic transplantation. Long-term monitoring of healthy PBSC donors remains important to guarantee the safety standards of bone marrow and peripheral blood stem cell mobilization and collection, including comprehensive medical examination before mobilization and collection, careful manipulation during collection, long term follow up after collection and so on.
Adolescent ; Adult ; Blood Donors ; Blood Specimen Collection ; adverse effects ; methods ; Female ; Hematopoietic Stem Cell Mobilization ; methods ; Hematopoietic Stem Cell Transplantation ; adverse effects ; methods ; Humans ; Male ; Middle Aged ; Transplantation, Homologous ; Young Adult

Maternal alcohol abuse is thought to be the common cause of mental retardation. Even moderate maternal alcohol consumption may produce fetal alcohol effects with behavioral and learning difficulties, if the drinking is associated with malnutrition. Especially, continuous alcohol consumption during critical period of brain development is very likely to produce fetal alcohol effects. The aims of this study are to investigate whether exogenous thyroxine treatment to alcohol -fed dams may ameliorate the detrimental effects of alcohol on the postnatal development of BDNF -containing Purkinje cell of the cerebellar cortex of the offspring. The morphological features of the growth and maturation were observed at 0, 7, 14, 21, 28 postnatal days via immunohistochemistry. In addition, electron microscopic finding of BDNF -containing Purkinje cell at P14 was also examined. Time -pregnant rats were divided into three groups. Alcohol -fed group received 35 calories of liquid alcohol diet daily from gestation day 6; control pair -fed group was fed a liquid diet in which dextrin replaced alcohol isocalorically; alcohol +/-T4 group received 35 calories liquid alcohol diet and exogenous thyroxine subcutaneously. As a result, a similar developmental pattern of BDNF -immunoreactive Purkinje cells was observed in control pair - fed and alcohol+/-T4 group on and after P14. These cells of alcohol -fed group showed immature features. Single -layer arrangement of these cells in alcohol -fed group was not completely achieved throughout postnatal life. Electron microscopic observations of BDNF -immunoreactive Purkinje cells at P14 revealed large nucleus, small cytoplasm, small amount of ribosomal collection and rudimentary cytoplasmic organelles in alcohol -fed group. The morphology of BDNF -immunoreactive Purkinje cell in alcohol +/-T4 group was similar to that in control pair -fed group. It was characterized by numerous short segments of rough endoplasmic reticulum, many of which showed a tendency of parallel alignment that suggested an attempt at Nissl body configuration. The cytology of Golgi complexes was also found within the cytoplasm in perinuclear location. Those observed differences of postnatal maturation patterns between alcohol -fed and alcohol +/-T4 group may indicate the beneficial effects on the postnatal development of BDNF -containing Purkinje cells in cerebellar cortex in the pups of thyroxine -treated alcohol -exposed dams. These results suggest that the increase of BDNF synthesis during early postnatal life caused by maternal administration of exogenous thyroxine may ameliorate fetal alcohol effects as a result of the dysthyroid state following maternal alcohol abuse.
Alcohol Drinking ; Alcoholism ; Animals ; Brain ; Brain-Derived Neurotrophic Factor ; Cerebellar Cortex ; Cerebellum* ; Critical Period (Psychology) ; Cytoplasm ; Diet ; Drinking ; Endoplasmic Reticulum, Rough ; Golgi Apparatus ; Immunohistochemistry ; Intellectual Disability ; Learning ; Malnutrition ; Organelles ; Pregnancy ; Purkinje Cells ; Rats* ; Thyroxine*

OBJECTIVETo demonstrate the effectiveness of a retrovirus-based plasmid vector coupled with nanometer material-polyamidoamine (PAMAM) dendrimer in stable gene expression of FVIII in vitro and to study the cytotoxicity of PAMAM.

METHODSThe retrovirus-based plasmid vector pLNC-FVIII BD was generated by cloning a B-domain-deleted (760aa - 1639aa) FVIII cDNA (FVIIIBD cDNA) into retroviral vector pLNCX. The complex that contained PAMAM and pLNC-FVIII BD transfer FVIII BD cDNA into NIH3T3 cell line. In day 2, 5, 10, 15, 30 after transferring, the antigen and procoagulant activity of human FVIII in the cell culture medium were measured by ELISA assay and one-stage method, respectively. RT-PCR was performed for the detection of FVIII BD mRNA. Inhibitory percentage of cell vitality was used for cytotoxicity of PAMAM.

RESULTSHuman FVIII was expressed for 30 days by transfected cells. The mean procoagulant activity of secreted FVIII in these 30 days was 0.929 U/ml, and the FVIII antigen was 0.188 micro g/ml by 10(6) cells in 24 hours, respectively. The level of FVIII didn't significantly decreased during these days. Inhibitory percent of cell vitality was only 5.32%.

CONCLUSIONPAMAM could effectively transfer pLNC-FVIII BD into NIH3T3 cells and FVIII could be stably and effectively expressed by the transfected cells. Cytotoxicity of PAMAM was low.

Animals ; Dendrimers ; Factor VIII ; genetics ; Genetic Vectors ; genetics ; Mice ; NIH 3T3 Cells ; Plasmids ; Polyamines ; pharmacology ; Retroviridae ; genetics