OBJECTIVETo explore the feasibility of evaluating complete ischemia-reperfusion injury (IRI) of the testis by contrast-enhanced ultrasonography with microbubbles (MB) targeted to P-selectin (MBp) in rabbits.

METHODSWe randomly divided 30 healthy adult rabbits into five groups of equal number (control, 0.5 h IRI, 1 h IRI, 2 h IRI, and 4 h IRI), prepared phospholipid MB and MBp, and performed contrast-enhanced ultrasonography of the bilateral testes with MB or MBp at an interval of 20 min at different times after IRI. When MB or MBp disappeared completely in the healthy testis at 4 to 5 min after intravenous injection, we recorded the power of the first frame (F-P) in the IRI testes followed by immunohistochemical staining of the testis tissue.

RESULTSCEU with MBp achieved a significantly higher F-P than that with MB in all the IRI groups (P < 0.05), which was (8.34 +/- 1.20) versus (1.87 +/- 0.25) 10(-5) AU at 2 hours, but there was no significant difference between MB and MBp in the control rabbits (0 AU, P > 0.05). Immunohistochemistry showed a significantly time-dependent increase in the expression of P-selectin in the vascular endothelial cells of the IRI testes, but not in those of the control.

CONCLUSIONContrast-enhanced ultrasonography with MBp can be used to evaluate the inflammatory reaction of testicular ischemia-reperfusion injury.

Animals ; Antibodies ; Disease Models, Animal ; Male ; Microbubbles ; P-Selectin ; immunology ; Rabbits ; Reperfusion Injury ; diagnostic imaging ; Testis ; blood supply ; Ultrasonography

OBJECTIVETo evaluate serum Nkx2-1 (NKX homeobox-1) levels in diagnosis of primary lung cancer.

METHODSThe serum NKX2-1 and CEA (carcinoma embryonic antigen) levels were measured in 61 patients with primary lung cancer admitted from May 2009 to December 2010 and 49 healthy individuals served as controls. The receiver operating characteristic curve (ROC) of NKX2-1 in diagnosis for primary lung cancer was analyzed. The value of serum NKX2-1 in diagnosing primary lung cancer was compared with that of CEA by X(2) test and Kappa test.

RESULTSThe serum Nkx2-1 levels in lung cancer were significantly higher than those in controls [(1.4206 ±0.1257)ng/ml compared with (0.7646 ±0.0734)ng/ml,P<0.01]. ROC analysis showed the area under the curve of serum NKX2-1 was 0.859. The Kappa value of NKX2-1 was higher than that of CEA (0.586 compared with 0.396,P<0.05). Combination of serum NKX2-1 with CEA improved the Kappa value to 0.704, and also had high sensitivity (83.6%) and specificity (87.0%) for diagnosis of primary lung cancer.

CONCLUSIONSerum NKX2-1 protein can be used as a marker for diagnosis of lung cancer, the combination of NKX2-1 with CEA may further improve the diagnostic value.

Biomarkers, Tumor ; blood ; Carcinoembryonic Antigen ; blood ; Case-Control Studies ; Humans ; Lung Neoplasms ; blood ; diagnosis ; Nuclear Proteins ; blood ; Sensitivity and Specificity ; Thyroid Nuclear Factor 1 ; Transcription Factors ; blood

OBJECTIVEMost of the therapeutic strategies for systemic inflammatory response syndrome (SIRS) is not effective. This study was to investigate the effect of continuous veno-venous hemodiafiltration (CVVHDF) on SIRS induced by cecum perforate peritonitis in piglets.

METHODSTwelve piglets (weighing 7-9 kg) were randomly divided into two groups: control and CVVHDF (n=6). The piglets of both groups were subjected to a cecum puncture to induce peritonitis which caused SIRS. After SIRS occurred the piglets of the CVVHDF group immediately received the CVVHDF therapy for 6 hrs, with a blood flow rate of 20 mL/min, a replacement rate of 300 mL/h, and a dialysis rate of 600 mL/h. The heart rate (HR), mean artery blood pressure (MABP), respiratory rate (RR), arterial blood gas analysis and blood cells count were measured and recorded at baseline and onset of SIRS, and 2, 4 and 6 hrs after SIRS occurred.

RESULTSWhen SIRS occurred, the HR and RR increased and the MABP, artery oxygen pressure (PaO2) and the count of white cells decreased in both groups. The HR of the CVVHDF group decreased significantly at 2 hrs (P < 0.05) and remained lower until 6 hrs after CVVHDF therapy (P < 0.01) compared with that of the control group. The RR of the CVVHDF group was significantly lower than that of the control group 6 hrs after CVVHDF therapy (P < 0.05). The MABP of the CVVHDF group increased significantly 4 and 6 hrs after therapy compared with that of the control group (P < 0.01, P < 0.05 respectively). There were no significant differences in temperature, PaO2 and blood cells count between the two groups during the experiment.

CONCLUSIONSCVVHDF has a positive effect on hemodynamics in piglets with SIRS induced by cecum perforate peritonitis.

Animals ; Blood Pressure ; Body Temperature ; Carbon Dioxide ; blood ; Cecum ; injuries ; Central Venous Pressure ; Female ; Heart Rate ; Hemodiafiltration ; Intestinal Perforation ; complications ; Male ; Oxygen ; blood ; Peritonitis ; complications ; Swine ; Systemic Inflammatory Response Syndrome ; therapy

OBJECTIVESince continuous blood purification (CBP) has the effects of eliminating inflammatory mediators and improving organs function, CBP had been applied to treat non-renal diseases for nearly 10 years, but few studies have been conducted in children with sepsis and multiorgan dysfunction syndrome (MODS), especially in China. The present study aimed to evaluate the clinical effect of CBP in treatment of children with severe sepsis and MODS.

METHODSTwenty-two children with severe sepsis and MODS admitted to our PICU from Aug. 2003 to Aug. 2005 were treated with continuous veno-venous hemodialysis filtration. Their heart rate, arterial blood pressure, doses of vasoactive agents, spontaneous respiratory rate, PO2/FiO2 and prognosis were investigated.

RESULTSCatheterization and CBP were carried out in all the 22 children. Continuous vein-vein hemodialysis filtration (CVVHDF) and pre-dilution were chosen. The duration of CBP was (64.4 +/- 34.5) h. All the children had tachycardia before CBP and the heart rate fell gradually to 45 +/- 13 bpm 4 h after CBP. Blood pressure (BP) was stable in 7 children without shock during CBP. Ten children with early shock could maintain normal BP during CBP, but the doses of vasoactive agents were tapered 1 to 5 h after beginning of CBP and use of these agents was discontinued at 2 to 8 h. BP was elevated by (25.2 +/- 10.7) mmHg (1 mmHg = 0.133 kPa) in 5 refractorily shocked children 4 h after CBP and returned to normal level 8 h later. The doses of the vasoactive drugs were reduced at 2 to 8 h and ended 4 to 16 h later, which was longer than that of children with early stage shock. The accelerated spontaneous respiratory rate was slowed down by 7 +/- 4 per minute 4 h later, PO2/FiO2 rose from (177.7 +/- 53.1) mmHg before CBP to (341.0 +/- 60.2) mmHg 4 h after CBP in children with respiratory failure and reached the normal value (5.3 +/- 2.1) h later. FiO2 declined to less than 50%. Pediatric critical illness score was 62.2 +/- 7.4 on admission and elevated to (86.6 +/- 9.0) 24 h later, which was a significant elevation as compared to that of children with sepsis who were not treated with CBP seen between Aug. 2001 and July 2003. The survival rate was 72.7% after CBP and the effective rate of the treatment was 90.9%, but was 36% in children who were not treated with CVVHDF.

CONCLUSIONCBP can effectively improve the vital organ's function of children with sepsis and MODS and raise their survival rate. Replacement fluid of modified Ports formula was useful for stability of serum potassium and sodium, but resulted in elevation of serum glucose, calcium, and osmolarity. The application of CBP in children with sepsis can lead to slight drop of blood pressure at the beginning and to bleeding during CBP.

Adolescent ; Blood Pressure ; Child ; Child, Preschool ; Female ; Heart Rate ; Hemofiltration ; methods ; Humans ; Infant ; Infant, Newborn ; Intensive Care Units, Pediatric ; Male ; Multiple Organ Failure ; etiology ; mortality ; physiopathology ; therapy ; Prognosis ; Sepsis ; complications ; mortality ; physiopathology ; therapy ; Severity of Illness Index ; Survival Rate ; Treatment Outcome

OBJECTIVETo investigate the effect of continuous veno-venous hemodiafiltration (CVVHDF) on endotoxin-induced acute lung injury (ALI) of piglets.

METHODSEighteen piglets were randomly divided into three groups: control group (n = 6); heparin group (n = 6) and CVVHDF treatment group (n = 6). All the animals were anesthetized by muscle injection of ketamine (30 mg/kg), then placed in supine position, received continuous intravenous infusion of ketamine with the rate of 10 mg/(kgxh). After placing a 4.5 cm (inner diameter) tracheal tube via tracheostoma, controlled mechanical ventilation was established using the assisted-controlled ventilation option of the NEWPORT 200. Respiratory rate at 30 breath/min; PIP at 10 cm H2O (1 cm H2O = 0.098 kPa); PEEP at 2 cm H2O and fraction of inspired oxygen at 0.3. A vein catheter was placed into right vena jugularis interna to administer a Ringer's solution. Initially, at a rate of 10 ml/kg, followed by a rate of 15 ml/kg when the mean arterial blood pressure was below 70 mm Hg (1 mm Hg = 0.133 kPa), the rate of 20 ml/kg was used when the mean arterial blood pressure was below 60 mm Hg. An 8Fr double-lumen catheter was inserted into left femoral vein and served as the pathway for CVVHDF. A Pulsiocath Pcco catheter was positioned into left femoral artery to monitor the circulatory parameters. All catheters were flushed with heparinized saline to prevent clotting. Then all the animals were given intravenous infusion of 150 microg/kg endotoxin within 30 minutes to induce ALI. When the oxygenation index < 300 and pulmonary compliance < 30% of the baseline, the animals of heparin group received heparin infusion to maintain blood active coagulation time (ACT) 180 - 250 s, the animals of treatment group received CVVHDF with the blood flow of 50 ml/min, replacement rate of 300 ml/h, dialysis rate of 600 ml/h and the ultrafiltrate rate of 350 ml/h for six hours, heparin infusion to keep blood ACT 180 - 250 s. The circulatory parameters: heart rate (HR), mean arterial blood pressure (MABP), central venous pressure (CVP), pulse contour cardiac output index (PCCI); systemic venous resistance index (SVRI), cardiac function index (CFI), external venous lung water index (EVLWI), left ventricular contractile index (dPmx); respiratory parameters: respiratory rate (RR), pulmonary compliance (Cdyn) were monitored; arterial blood gas analysis was performed and oxygenation index (PaO2/FiO2) was calculated. All the parameters were recorded at baseline (B), onset of ALI (A 0 h), two hours (A 2 h), four hours (A 4 h), six hours (A 6 h) after ALI.

RESULTSNo significant difference in circulatory parameters, respiratory parameters and blood gas analysis were found at B and A 0 h among the three groups. When the ALI occurred, PaO2/FiO2, Cdyn, MABP and PCCI of the three groups decreased; HR, RR, EVLWI, SVRI increased. After four hours of ALI, the RR, EVLWI, SVRI, CFI and dPmx of treatment group were improved, the differences were significant compared with the other two groups (P < 0.05). After six hours of ALI, the HR, PCCI, MABP, PaO2/FiO2 and Cdyn of treatment group were significantly improved, compared with control group and heparin group (P < 0.05). There were no significant differences in any of the parameters between control group and heparin group. The difference in CVP among three groups was not significant.

CONCLUSIONCVVHDF has a good effect on hemodynamics of the endotoxin-induced ALI of the piglets.

Acute Lung Injury ; etiology ; physiopathology ; therapy ; Animals ; Endotoxins ; adverse effects ; Hemodiafiltration ; Hemodynamics ; Swine

Female ; Humans ; Middle Aged ; Multiple Myeloma ; complications ; Pulmonary Embolism ; etiology ; pathology ; Venous Thrombosis ; etiology

OBJECTIVEPractice recommendations have evolved, and consensus now exists among leading organizations such as the American College of Critical Care Medicine (ACCM) and Surviving Sepsis Campaign that fluid infusion is best initiated with boluses of 20 ml/kg, commonly requires 40-60 ml/kg but can be as much as 200 ml/kg if the liver is not enlarged and/or rales are not heard. The present study aimed to investigate and compare the changes of the hemodynamics and extravascular lung water after higher volume fluid resuscitation in a piglet model of endotoxic shock.

METHODTwenty piglets were used for establishing animal models of endotoxic shock by intravenous infusing lipopolysaccharide (LPS). The experimental animals were divided into three groups according to the volume infused during the resuscitation. The three groups received different volume of saline in less than an hour after endotoxic shock. By the PiCCO plus system, we investigated the changes of hemodynamics and extravascular lung water.

RESULTAfter fluid resuscitation, global end diastolic volume inder, (GEDI) and intrathoracic blood volume index, (ITBI) markedly increased in the group of 80 ml/kg and 120 ml/kg, but there was no change in the group of 40 ml/kg. GEDI: Fifteen min after fluid resuscitation R1 was (261 ± 64) ml/m(2), R2 (457 ± 124) ml/m(2), R3 (413 ± 148) ml/m(2), 4 h R1 (251 ± 68) ml/m(2), R2 (422 ± 70) ml/m(2), R3 (470 ± 160) ml/m(2); ITBI: Fifteen min after fluid resuscitation R1 was (335 ± 69) ml/m(2), R2 (550 ± 179) ml/m(2), R3 (520 ± 183) ml/m(2), 4 h R1 (314 ± 84) ml/m(2), R2 (534 ± 96) ml/m(2), R3 (594 ± 200) ml/m(2) (R1 vs. R2 vs. R3, F = 26.373, P < 0.05; R1 vs. R2, R1 vs. R3, P < 0.05; R2 vs. R3, P > 0.05). CI of all three groups significantly decreased when the models were established. After fluid resuscitation, the base level was maintained in the group of 80 ml/kg and 120 ml/kg, but it was under the basic level in the group of 40 ml/kg.Fifteen min after fluid resuscitation R1 was (4.5 ± 0.7) L/(min·m(2)), R2 (6.4 ± 2.2) L/(min·m(2)), R3 (5.5 ± 0.7) L/(min·m(2)), 4 h R1 (4.1 ± 1.0) L/(min·m(2)), R2 (5.2 ± 0.9) L/(min·m(2)), R3 (5.1 ± 0.8) L/(min·m(2)). There was no significant difference in CI between these two groups (P > 0.05).ELWI of the group of 80 ml/kg and 120 ml/kg were still higher than that of the group of 40 ml/kg, 15 min after fluid resuscitation R1 was (19.2 ± 8.6) ml/kg, R2 (29.2 ± 5.5) ml/kg, R3 (23.4 ± 8.2) ml/kg, 4 h R1 (18.3 ± 6.5) ml/kg, R2 (23.8 ± 2.6) ml/kg, R3 (21.4 ± 3.9) ml/kg, but there was no significant difference in ELWI among the groups (P > 0.05).

CONCLUSIONResuscitation with higher volume of fluid infusion in the early stage of endotoxic shock was more efficient to increase the preload and maintain the cardiac output at the baseline level, and might reduce the need for vasoactive agents. Meanwhile, resuscitation with higher volume of fluid in the early stage of endotoxic shock did not sharply increase the extravascular lung water.

Animals ; Blood Volume ; Central Venous Pressure ; Disease Models, Animal ; Extravascular Lung Water ; Female ; Fluid Therapy ; methods ; Hemodynamics ; Lung ; metabolism ; physiopathology ; Male ; Random Allocation ; Resuscitation ; methods ; Shock, Septic ; metabolism ; physiopathology ; therapy ; Sodium Chloride ; administration & dosage ; therapeutic use ; Swine

OBJECTIVETo identify the volatile organic compounds (VOCs) in lung cancer tissue and lung cancer cell lines.

METHODSThe lung cancer tissue samples from 18 patients were cultured and 4 lung cell lines (A549, NCI-H446, SK-MES-1, BEAS-2B) were also included in the study. Air samples in the headspace of culture flasks were analyzed for VOCs with solid-phase micro-extraction and gas chromatography-mass spectroscopy technique (SPME-GC/MS).

RESULTTwo kinds of VOCs 2-pentadecanone and nonadecane were detected in lung cancer cell lines A549, NCI-H446 and SK-MES-1. The concentration of 2-pentadecanone were (1.382 + or -0.171) X 10(-5)mg/L, (1.681 + or - 0.190) X 10(-4)mg/L and (2.835 + or - 0.401) X 10(-6)mg/L, respectively; the concentrations of nonadecane were (8.382 + or - 0.606 ) X 10(-6)mg/L, (1.845 + or - 0.130) X 10(-5)mg/L and (6.220 + or - 0.362) X 10(-6)mg/L), respectively. The eicosane was detected in A549 and NCI-H446 with the concentration of (8.313 + or - 1.130) X 10(-6)mg/L and (1.020 + or - 0.141) X 10(-5)mg/L), respectively. All the 3 VOCs were not detected in cell line BEAS-2B. The concentrations of 12 VOCs including decane, 2- pentadecanone, nonadecane and eicosane were high in 18 lung cancer tissue samples; the concentrations of 2-pentadecanone were 5.421 X 10(-6)mg/L-3.621 X 10(-5)mg/L,those of nonadecane were 5.805 X 10(-6)mg/L-1.830 X 10(-5)mg/L, those of eicosane were 2.730 X 10(-6)mg/L-2.343 X 10(-5)mg/L. There were no differences of VOCs levels among patients with different cancer differentiation (P>0.05). The concentration of eicosane in the non-squamous carcinoma was higher than that in squamous carcinoma, the same results were confirmed in the lung cancer cell lines.

CONCLUSIONThis study has identified VOCs produced by lung cancer tissue, which may support to use breath test as a complementary noninvasive diagnostic method for lung cancer.

Adult ; Aged ; Alkanes ; metabolism ; Biomarkers, Tumor ; metabolism ; Cell Line, Tumor ; Female ; Humans ; Lung Neoplasms ; diagnosis ; metabolism ; Male ; Middle Aged ; Tumor Cells, Cultured ; Volatile Organic Compounds ; metabolism

The nucleotide sequence and N-, C-terminal amino acid sequences of alpha,beta-subunit of glutaryl 7-ACA acylase C130 from Pseudomonas sp. 130 were determined. The alignment of the acylase C130 with the other acylases shows that it has high homology with the acylases from Pseudomonas sp. GK16 and C427, but low homology with the others. There is large difference in the N-terminal of alpha-subunit, while the N-terminal of beta-subunit has significant conservation.
Amino Acid Sequence ; Base Sequence ; DNA, Bacterial ; analysis ; Genes, Bacterial ; Molecular Sequence Data ; Penicillin Amidase ; genetics ; Pseudomonas ; enzymology ; genetics ; Sequence Homology, Amino Acid ; Sequence Homology, Nucleic Acid

Objective:To explore the function of tumor derived IgG (tIgG) and whether the tIgG can inhibit T cells activity.Methods:The tIgG was purified from ovarian cancer tissue.The cord blood monocyte cells (CBMC) and cord blood lymphocyte (CBL) were isolate from human umbilical cord blood.The CBMC and CBL were stimulated with phytohaemagg lutinin (PHA) in order to let the CBMC and CBL in the state of proliferation.Carboxyfluorescein succinimidyl amino ester (CFSE) was cultured with CBMC and CBL.CFSE had no cell toxicity,which could penetrate through the cell membrane and combine the intracellular protein.The fluorescence intensity decreased with the proliferation of cells step by step,so the proliferation of these cells could be detected in flow ctytometry.The tlgG which was puri fied from ovarian cancer tissue was divided into three groups,1 mg/L group,10 mg/L group,and 100 mg/L group,and the intravenous immunoglobulin (IVIG) was also divided into three groups too.The CBMC and CBL were treated by tIgG with 1 mg/L,10 mg/L,and 100 mg/L in order to observe the proliferation of T cells.The cells were treated with IVIG as a positive control group,and the cells were treated with phosphate buffer saline (PBS) as a negative control.The proliferation of CD4 + or CD8 + T cells were detected in CBMC and CBL.The proliferation of the T cells in CBMC and CBL after 64 h and 86 h were detected.Results:In the system of CBMC,the tIgG could suppress the proliferation of CD4 + or CD8 + T cells.The results could also be found in the system of CBL.The CD4 + or CD8 + T cells in the group which were treated with PBS were more active than those in the group which were treated with tIgG and IVIG.The suppression in the group which were treated with tIgG,was stronger than that in the group treated with IVIG.In addition,the suppression of T cells in the group which were stimulated with tIgG as 100 mg/L was more effective than that in the group which were stimulated with tIgG as 10 mg/L.This could prove that tIgG had the function of immunomodulation.Conclusion:The tIgG can be involved in immune escape of cancer.