A method for isolation and primary culture of guinea pig alveolar type Ⅱ epithelial cells is reported. The injurious effects of phospholipase A2 (PLA2) on the cultured cells were studied. Trypsinizing solution was instilled into the bronchoalveolar space. The lungs were digested, sectioned, vortexed and filtered. The yield of original cell mixture was 172.3?106 cells/guinea pig, of which 33.6% were type Ⅱcells. At 48 h of culture following short term selective attachment 7.9?106 cells were obtained, of which 66.1% were type Ⅱ ceils. The purity could further be raised to 76.2% (63.9% -92%) if pretreated with ethyienediarnine tetraacetic acid (EDTA) before trypsinization to remove attached cells. The characteristics of cultured cells were observed under phase contrast microscopy, light microscopy and electron microscopy. After exposure of the cells to 300U/ml PLA2 for 4h at 37℃, detachment and lactic dehydrogenase (LDH) release of the cultured cells increased significantly. It is suggested that PLA, may damage and promote detachment of cultured type Ⅱ cells and it may play an important part in the development of some diseases.

We investigated whether platelet-activating factor (PAF) mediates endotoxin-induced changes of lipoperoxidation, lysosomal enzyme release and increased extravascular lung water content in dogs using a specific PAF receptor antagonist, SRI 63-441. Endotoxin infusion caused an increase in serum malonyldialdehyde (MDA) from baseline (100%) to 130.04?14.00% and 169.16?32.49% and ?-glucuronidase (?-g) activity to 191.05?86.71% and 242.54?49.09% at 3 and 6h, respectively. The bronchoalveolar lavage fluid (BALF) ?-g activity, lung MDA and extravascular lung water also significantly increased (P

The effects of hydrogen peroxide (H2O2) on endothelial-polymorphonuclear cells (EC-PMN) adhesion and their mechanisms wsre studied in cultured bovine pulmonary artery endothelial monolayers in vitro. H2O2 at various concentrations (10-1, 10-2, 10-3mol/L respectively) stimulated EC dependent PMN adhesion, of which l02mol/L H2O2 was the most potent one, increasing adhesion to 2.3 times that of the control. Pretreatment of PMNs with SRI 63-441, a platelet-activating factor (PAF) receptor antagonist, had no effect on H2O2 induced EC-PMN adhesion. Pretreatment of ECs with SRI 63-441 before H2O2 exposure significantly decreased PMN adherence to ECs. Pretreatment of ECs with phospholipase A2 inhibitor p-bromophenacyl-bromide or cahnodulin antagonist chlorpromazine and aildum ion chelate EGTA obviously decreased H2O2 induced increment of EC-PMN adhesion. These results suggest that H2O2 may activate ECs, causing the inflow of extracellular calcium or the release of calcium from intracellular deposits. Increased intracellular Ca2+ may bind with calmodulin to activate phospholipase A2 thus initiating PAF synthesis and promoting EC-PMN adhesion.

ObjectiveTo investigate the effects of controlled subcutaneous analgesia(PCSA) with sufentanil in patients underwent abdominal surgery and the occurrence of adverse reactions. Methods90 patients underwent abdominal surgery were randomly divided intothree groups with 30 cases each. Group Ⅰ :sufentanil 0. 04μg · kg-1 ·h-1 for postoperative analgesia; Group Ⅱ : sufentanil 0. 06μg · kg-1 · h-1 for postoperative analgesia; Group Ⅲ :sufentanil 0.08μg/kg · h for postoperative analgesia;PCSA started at the end of operation with catheter buried in deltoid muscle. Visual analgesia scale(VAS) ,Rasmay scale and side effects were recorded at 4,8,16,24 and 48h after operation. ResultsVAS scores decreased significantly in group Ⅱ and group Ⅲ compared with that in group Ⅰ ( all P ＜ 0.05) ;There were no differences of Ramsay scores between groups ( all P ＞ 0. 05) ;There were three cases of respiratory depression in group Ⅲ. ConclusionPCSA with sufentanil (0.06μg/kg) had exact analgesic effects for postoperative abdominal surgery patients with less adverse reactions.

Objective To study the applicability of Vienna Test System ( VTS) to psychological screening of flying cadets,and to establish a corresponding standard model for flying cadets .Methods VTS and pilot psychological selection system were used to evaluate the mental health of 134 cases of flying cadets , SPSS 13.0 was used to compare their consistency and relevance.Results The results of VTS included the raw score of overall mean duration [(46.68 ±21.06)s] on 2hand test, the correct reaction times (246.54 ±25.88) on DT test, the normal values of overall mean duration [(27.84 ±21.98)s] on 2 hand test,and the correct reaction times (60.57 ±18.61) on DT test and other indexes. Pearson coefficient was calculated as predictive validation .The composite score of VTS was significantly positively correla-ted with the first platform(r=0.46,P<0.0001), the second platform(r=0.27,P<0.05) and the psychological perform-ance of the pilot psychological selection system (r=0.31,P<0.05).Kappa-test showed that the two methods were highly consistent(Kappa=0.53).Conclusion The application of VTS to screening the mental health of flying cadets can yield and quick but accurate results , which are of high reliability and validity .It is undoubtedly of great significance for improving the level of aviation security ,objectivity of selection results and predictive validity .

Objective To explore the relation of the expressions of MDM2 and VEGF in osteosarcoma with the pathological parameters and prognosis of the tumor.Methods The expressions of MDM2 and VEGF were detected with immunohistochemical(SP) method in specimens from 56 cases of osteosarcoma.The correlation between the expressions of MDM2,VEGF and pathological grade,metastasis and prognosis was analyzed statistically.while 8 cases of fibrous dysplasia of bone were used as negative control group.Results The positive rates of MDM2 and VEGF in osteosarcoma were 64.3%(36/56) and 67.9%(38/56),respectively .MDM2 and VEGF didn't express in negative control group.The expression of MDM2 and VEGF were not significantly correlated to the pathological grades of the osteosarcoma,but which were significantly correlated with tumor metastasis and prognosis(P

Intravenous administration of E. coli endotoxin (2mg/kg) caused a persistent increase in platelet activating factor (PAF) contents in arterial blood and bronchoalveolar fluid (BALF). Meanwhile phospholipase A2(PLA2) activity in serum and BALF was elevated and mean arterial blood pressure declined. The total cell number, expecially polymorphonuclear leukocytes and lymphocytes, and protein concentration in BALF were markedly increased 6 h after endotoxin injection. Lung index and extravascular lung water content of endotoxin-treated animals were significantly highter than those of controls. Pretreatment with PAF receptor antagonist SRI 63-441 blocked the increase in PLA, activity and attenuated endotoxin-induced hypotension and acute lung injury. The results suggest that PAF mediates endotoxin-induced lung injury, and leukocyte activation by PAF and the subsequent release of oxygen metabolites and lysoenzymes are important intermediate mechanisms leading to high permeability pulmonary edema.

We established a method to study vascular permeability in vitro on perfused endothelial cell monolayer cultured on micropore filter membrane. It can be used to determine filtration coefficient (Kf) and osmotic reflection coefficient (?) to proteins. Hanks balanced salt solution (HBSS) or 5 g/L albumin in HBSS was used to perfuse confluent endothelial monolayer. Control Kf values were 10.1 ?0.75 and 3.6?0.75?l . min-1 . cm-2 . kPa-1 (n = 3, x?sx) respectively for HBSS and albumin HBSS perfusion, suggesting that albumin may decrease endothelial monolayer permeability to water and small molecules. After exposure of endothelial monolayer to 10-8 mol/L platelet-activating factor (PAF) for 30 min, Kf values increased to 193.1% and 133.3% respectively for HBSS and albumin HBSS perfusion. Protein clearance rate (?l. min-1 . cm-2:) and osmotic reflection coefficient of control endothelial monolayer were 8.0?3.22 and 0. 37?0.09 respectively. In PAF treated endothelial monolayer,they were 12.2 ? 2.95ul min-1 cm 2 and 0.18?0.06, revealing increased permeability to albumin. Computer-assisted image processing demonstrated that PAF treatment decreased cell area while increased cell form factor and intercellular space. The results sug-gest that endothelial cells retracted, rounded and it may be an important mechanism in PAF-induced increased vascular permeability.

Objective To explore the effects of exercise on the levels of neurotrophin-3 (NT-3) in the skeletal muscles of streptozotocin-induced diabetic rats. Methods The rats were divided into an 8-week exercise group (A), a 4-week exercise group (B), a diabetes control group (C), an exercise group (D) in which no dia-betes was induced, and a control group (E). The rat model of diabetes was induced in the rats of groups A, B and C by intra-abdominal injection of streptozotocin (STZ) at 55 mg/kg. The exercising rats were forced to swim for 60 minutes once daily, 5 days a week. The levels of NT-3 in skeletal muscles were measured with an enzyme-linked immunosorbent assay. Cadual nerve conduction velocity (CNCV) in all of the rats was evaluated at the beginning, and after 4 and 8 weeks of swimming exercise. Results NT-3 levels in the skeletal muscles in group C were sig-nificantly lower than in groups A, D and E. There was no statistically significant difference in NT-3 levels between groups B and C. The NT-3 levels showed a significant positive correlation with CNCV at the 8th week. Conclu-sions The increase in NT-3 levels of skeletal muscles induced by exercise could contribute partially to the im-provement of diabetic neuropathy.

ObjectiveTo investigate the size effect of hydroxyapatite nanoparticles on proliferation and apoptosis of osteoblast-like cells.MethodsCetyltrimethylammnonium bromide (CTAB) was used to regulate the size of nano hydroxyapatite (nHAP) particles.All obtained particles were characterized by transmission electron microscopy (TEM),X-ray diffraction,dynamic light scattering and chemical analysis.HAP films were obtained by slowly coating cover glasses with 1％ HAP particle suspension.MG-63 cells on three different films(20HAP,40HAP and 80HAP) were cocultured for up to 5 days.Cell proliferation assay was obtained by methyl thiazolyl tetrazolium (MTT).Cell apoptosis was detected by flow cytometric detection.Cell ultrastructure morphology was observed by TEM observation.ResultsnHAP with diameter of 20 nm,40 nm and 80 nm were synthesized and and analyzed.The MG-63 cells were cultured on three different fihns.The optical density value of cells on 20HAP was 1.22±0.13 after 5 days incubation,and there was no different compared to the control group(F=6.843,P=0.124).Cell number and viability were significantly higher on 20HAP compared to large nHAP after 5 days incubation.The percentage of apoptotic cells increased with increasiug nHAP particle size.TEM images showed 20HAP was found in cytoplasm and cell morphology had no changes.ConclusionBoth cell proliferation and cell apoptosis are related to the size of the nHAP particles.20HAP was the most effective on promoting cell growth and inhibiting cell apoptosis.