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.

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 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 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 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 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.

Objective To obtain the expression pattern of imprint gene H19 in JEG-3 cell in order to explore the regulation mechanism of H19 on trophoblast cellular biological behavior .Methods After correct identification with sequencing for the recom-binant eukaryotic expression plasmid pRc/CMV which including the whole length of H19 cDNA ,the plasmid was transfected to the cell line JEG-3 .The expression of H19 mRNA was observed and the gene expression profile of three groups of JEG-3 cell were de-tected with Affymetri :U133 plus 2 .0 Array .Results After being transfected with target H 19 gene ,the expression of the mRNA level was significantly increased compared with control group .And the gene expression profile was changed significantly .19 genes were up-regulated ,77 genes were down-regulated .Expression levels of HES1 gene which being choosed as a different expression gene were detected by fluorescence quantitative PCR in severe preeclampsia placenta tissue and normal late pregnant placenta .The expression level of HES1 mRNA in severe preeclampsia placenta decreased significantly than normal late pregnant placenta tissues . Conclusion Many genes induced by H19 have been screened by high-throughput gene chip method .It provides the experimental ba-sis for advanced studying the regulation the cellular biological behavior with H 19 gene .

Background Retinal ganglion cell (RGCs) death following ischaemic insult is the major cause of a number of vision-threatening diseases.Recent studies confirmed that micro RNA (miR-30b) can alleviate hypoxy-induced cardiac injury.However,whether miR-30b can protect RGCs against oxygen-glucose deprivation damage is still not ellucidated.Objective The aim of this study was to investigate the protective effect of miR-30b on RGCs damage caused by oxygen-glucose deprivation.Methods The retinas were isolated from the eyeballs of eight SD rats aged postnatal 24 hours and RGCs were primarily cultured.The cells were divided into the recombinant adeno-associated virus (rAVV) control group,rAAV-miR-30b mimic group and AAV-miR-30b inhibitor group.Then the cells were transfected using rAVV-miR plasmid,rAAV-miR-30b mimic plasmid and AAV-miR-30b inhibitor plasmid,respectively for 6 days with the RGCs ∶ AAV as 1 ∶ 10 000.The cells were cultured with low glucose medium in hypoxygen incubator (5％ CO2,17％ N2,3％ O2) or 5％ CO2 incubator respectively for 24 hours.Cell viability was detected by cell counting kit-8 assay.The expression of Tubulin Ⅲ,a neuron specific marker,was detected by immunofluorescence technology to evaluate the survival of RGCs.The apoptosis and necrosis of the cells were assessed by Hoechst/PI double staining.Results The RGCs grew well with round shape and 1 3 processes 7 days after cultured in the normal cells.However,the RGCs were diminished and the cell process disrupted in the oxygen-glucose deprivation group.The relative vability of the cells was 3.310-±0.162 in the rAAV-miR-30b mimic group,which was significantly higher than 0.949±0.141 in the rAAV-miR-30b inhibitor group and 0.900±0.181 in the rAAV-miR control group(t=10.508,10.296,both at P＜0.001).It was positively expressed in survival RGCs,with the red fluorescence.The number of Tubulin Ⅲ+ cells was (13.800± 1.924)/field in the rAAV-miR-30b mimic group,showing a significant increase in comparison with (0.600±0.548)/field in the rAAV-miR-30b inhibitor group and (0.800± 1.304)/field in the rAAV-miR control group (t =15.141,14.912,both at P ＜ 0.001).Significant differences were found in the apoptosis rate and necrosis rate among the rAAV-miR-30b mimic group,rAAV-miR control group and PBS group (F=10.851,P=0.002;F=6.378,P=0.013),and the apoptosis rate and necrosis rate in the rAAV-miR-30b mimic group were considerably lower than those in the rAAV-miR control group and PBS group (all at P＜0.05).Conclusions The oxygen-glucose deprivation models can be established in RGCs by hypooxygic and low-glucose cultivation.rAAV encoding miR-30b mimics transfection can protect RGCs against oxygen-glucose deprivation damage.