This study was designed to investigate the changes of the contents of throm-boxane (TXA2) and prostacyclin (PGI2) in the pulmonary tissue and in the plasma of the pulmonary arteries and left atrium of the rabbits during ARDS induced with oleic acid injection and the interrelationship between the two substances.The contents of TXA2 and PGI2 were determined indirectly,by measuring the amounts of TXB2 and 6-keto-PGF1a,the stable metabolites of TXA2 and PGI2 respectively,with radioimmunoassay.After an injection of oleic acid to the rabbits (0.045ml/kg),the contents of TXB2 in both the plasma and lung tissue were significantly elevated The content of 6-keto-PGF1a in the plasma significantly increased at 30 minutes and reached the highest point at 180 minutes after the injection,and that in the lung tissue began to increase at 5 minutes and reached the peak at 60 minutes after injection The ratio of TXB2/6-keto-PGI18 increased in the early stage after injection and then reduced both in the plasma and the lung tissue.The results indicate that the contents of TXA2 and PGI2 in the plasma are not always parallel with those in the lung tissue,which suggests that the variation of the ratio of TXA2/PGI2 may play an important role in the pathogenesis of ARDS.

Respiratory distress syndrome was produced in dogs with the combination of oleic acid injection and gunshot wounding,and the therapeutic effects of naloxone(NAD and positive end expiratory pressure ventilation(PEEP)were assessed with the examinations of macropatholo-gy of the lungs,lung water ratio.respiratory rate(RR)and blood gas analysis.It was found that RR was rapidly and significantly increased tPaO2 decreased and Qs/Qt increased and there was severe pulmonary hemorrhage,edema and atelectasis in the controls after injury.NAL treatment could slightly alleviate the increase of RR and prevent the early of PaCO2,but it had no effects on the changes of PaO2,Qs/Qt and increase pulmonary hemorrhage,edema and atelectasis.PEEP could improve the decrease of PaO2 and the increase of Qs/Qt and prevent pulmonary atelectasis and alveolar edema from occurring,but it could not significantly stop the increase of lung water ratio.

During the recent conflicts in Iraq and Afghanistan,traumatic brain injury (TBI)has become the most prev-alent military injury that is described as the signature injuryof the current military operations.It usually causes no or mild external injury but results in serious long-lasting neuropsychiatric abnormalities,which have far-reaching impact on veterans,their families and the American society.Here we describeol the investment in TBI from the US government and the development in the diagnosis and treatment of mild TBI on the battlefield before putting forward some proposals for the Chinese army.

OBJECTIVE A practical gene chip which aimed to detect and identify pathogens rapidly and exactly is developed on the basis of patent technology of nano-enlargement-detection. METHODS Oligonucleotide probes for the specific gene fragments of target pathogens were designed and immobilized on gene chip.Target sequences were labeled by nanogold as reporter materials.After hybridization,its results were recorded by the interaction between nanogold and silver which amplified the hybridization signal to form brown particles,which could be detected by naked eyes. RESULTS The probes designed were all of strong specificity and great reliability possessing identity of hybridization conditions.The reaction time for marking could be decreased by properly raising the ratio of nanogold and nucleic acid and the speed of labeling reaction could be fastened significantly by gentle agitation.A better hybridization results could be obtained when the samples were hybridized for 8 hours at 45℃ with 0.8 mol/L ionic strength,and then strictly rinsed.Furthermore,the hybridization efficiency could be increased remarkably by slight circumgyratation.A better chromatic effect resulted from the reaction way in 3min?3 at 37℃.The sensitivity of gene chip assays in this test could reach to 100 fmol/L.Compared with traditional detection approach,detection by the chip displayed such advantages as speediness and simplicity and the detection results could be easily recognized by naked eyes. CONCLUSIONS The chip detection technology has met the demand of design exhibiting high sensitivity,strong specificity,and easy operation without special device and showing a promising prospect.

Objective For realizing the simultaneous detection of multiple pathogens,by looking for a method with a combination of the new gene chip detection system based on nano-gold with the technology of restriction endonuclease without PCR.Methods Helicobacter pylori,Mycoplasma pneumoniae,Chlamydia trachomatis,Candida albicans,Ureaplasma urealyticum,and EB virus were selected as the experimental targets.Endonuclease Hha Ⅰ was selected as tool enzyme.After bering digested by Hha Ⅰ,the digested fragments of samples were tailed with poly-A.The samples were then detected by the gene chip detection system based on nano-gold.Both specific and common probes were used in the hybridization.The coincidence rate of the detection results between the new constructed chip test and the fluorescence quantitative PCR test in 168 clinical samples was examined.The stability and sensitivity of chips detection were also checked.Results The new constructed nano-gold-baesd gene chip combined with restrictive enzyme digestion without PCR could be used to detect the target pathogens.The coincidence rate of the chip detection test and fluorescence quantitative PCR test in 168 clinical samples was 89.2%.Chip detection results showed that the stability of chips detection was 100% and the sensitivity was 50pmol/L.Conclusion The newly constructed nano-gold-baesd gene chip combined with restrictive enzyme digestion without PCR can be widely applied in the simultaneous detection of Mycoplasma,Chlamydia,fungus,virus and bacteria.It shows a bright prospect in increasing the throughput of identifieation of pathogene.

OBJECTIVE To study the imprinting status of H19 gene in normal villi tissue during the first trimester,and its relation to the invasion of trophoblast. METHODS Using PCR-RFLP methods to examine the imprinting status of H19 gene in 93 cases of normal villi tissue during the first trimester. RESULTS Among 93 cases, heterozygous genotypes were found in 42 cases. And 11 cases of biallelic expression were found among these 42 cases of heterozygous genotypes from 5 to 9 weeks, however no biallelic expression existed from 10 to 12 weeks. CONCLUSIONS During the first trimester, H19 is expressed biallelically at the first 10 weeks. The H19 gene may dynamicly change in the trophoblast, and the dynamic change may have close relationship with the invasion of the trophoblast.

ObjectiveTo investigate the expression of paired immunoglobulin-like receptor B (PirB) in optic nerve, visual cortex, cerebella, spinal cord and sciatic nerve of normal adult mice.MethodsTwelve healthy adult BALB/c mice were randomly and equally divided into two groups.The immunohistochemistry and Western blot were used to detect the expression of PirB in the tissues described above respectively.ResultsBoth the immunohistochemistry and Western blot test revealed that the expression of PirB was positive in the optic nerve, visual cortex, cerebella and spinal cord, but negative in the sciatic nerve.The positive signals in the sections were located in the cell bodies and the neurites were observed in some of them.Western blot showed the apparent positive band of PirB in the optic nerve, visual cortex, cerebella and spinal cord rather than in the sciatic nerve.The protein expression level of PirB was relatively high in the visual cortex (P ＜0.05) but relatively low in the optic nerves (P ＜0.01).ConclusionThe PirB expresses positively in the optic nerve, indicating that PirB protein may closely correlate with the poor regeneration of the optic nerve.

OBJECTIVE To develop a preparation technique of sample of one-time PCR with common primers based on ribotyping which was combined with the detection system of nanogold-based gene chip to detect clinical bacterial pathogens.METHODS According to the highly conserved regions of rDNA,the common primers were designed and used to amplify each target bacterial ISRs by one-time PCR,and the specific oligonucleotide probes for each target ISRs were designed,utilized to establish the new nanogold-based gene chips.After the characteristics of the chip such as sensitivity,specificity and reliability were determined,the chip was used to detect clinical samples.RESULTS The designed common primers could amplify the 12 target bacteria successfully by one-time PCR.All selected probes were of strong specificity and great reliability.The chip had high sensitivity,specificity and reliability,reaching 50 fmol/L of detection sensibility.Clinical detection results showed the chip had a great accuracy.CONCLUSIONS Compared to multi-PCR chip detection,the detection procedure and complexity of the chip are decreased significantly,and have more practical value in clinical pathogens detection.

Objective To observe the variation of dynamic 64-slice CT perfusion imaging of rats with traumatic brain injury and discuss the relating pathophysiological basis.Methods A total of 80 adult male SD rats were randomly divided into three groups according to random number table,ie,normal control group,sham injury group and injury group.The injury group was divided into eight subgroups at time points of 2,6,12,24,48,72,120 and 168 hours.The detection of CT perfusion imaging,water content and blood-brain barrier permeability was done in the injured rats at all time points.The pathological changes were also observed to calculate their correlation with CT perfusion parameters of the injured region.Results The relative value of the blood perfusion was decreased significantly to the mimimum within 24 hours after injury.Within 2-12 hours,relative cerebral fluid(rCBF)and relative cerebral blood volume(rCBV)remained in a low perfusion state,with just a little increase.Relative mean transit time(rMTT)was prolonged and permeability surface(PS)increased.rCBF and rCBV were increased gradually with time,which was reversed till at 24 hours after injury and the injured side was in a high perfusion state,with the highest value of PS.The perfusion reached peak at 48 hours after injury and then became normal gradually.The water content was increased at 2 hours after injury and reached its peak at 48 hours.The permeability of blood-brain barrier(BBB)began to increase at 2 hours after injury and reached the peak at 24 hours.rCBF and rCBV were positively correlated with change of brain edema and PS was positively correlated with BBB permeability.Conclusion The dynamic 64-slice spiral CT perfusion imaging reflects the variation of BBB and edema and can be used as noninvasive imaging method for predicting the degree of brain perfusion and edema.

Objective To investigate the protecting mechanism of heat stress pretreatment on acute lung injury(ALI). Methods The oleic acid ALI mouse model was built to dynamically observe the binding capacity and the binding affinity of glucocorticoid receptor(GR),the levels of GR,heat shock protein 90(Hsp90)and Hsp70 before and after hyperthermic stress pretreatment. Results Heat stress pretreatment had significant protective effect on ALI.Western blotting showed insignificant changes of GR levels but progressive increase of level of Hsp70 and Hsp90.Heat stress pretreatment exerted insignificant effect on Bmax and Kd of GR,shown by radio ligand binding assay after ALI. Conclusion The protective effects of heat stress pretreatment on ALI of mouse may relate to its ability of keeping stable GR level and increasing levels of Hsp70 and Hsp90.