Objective To prepare the recombination protein and polyclonal antibodies against human apolipoprotein A5(ApoA5)and detect the expression of ApoA5 in human tissues with the antibody,then further study the fuction and serum level of ApoA5.Methods With PCR and gene recombinated techniques,the prokaryotic ApoA5 expression vector was constructed.The His-ApoA5 fusion protein was expressed in E.ColiM15,and the fusion protein was injected into New Zealand rabbits to prepare polyclonal antibody.The antibody was tested for their specificity with ELISA and Western-blot,then it was used to detect the tissue distribution of ApoA5.Results It was found that there was a specific brand at 40 ku of the recombination ApoA5(rApoA5);the polyclonal antibody had high immunoreactivity and specificity.ApoA5 was expressed in human sera and hepatic tissue,not in other tissues(heart,vessel,spleen,intestine and lung),regardless of the lipid level.Conclusion The pQE30-ApoA5 is constructed successfully,the rApoA5 can be expressed in E.Coli M15;ApoA5 antibody has highly specifity.It can be used in clinical test.There is ApoA5 in human serum,and ApoA5 is expressed in liver which may be significant in lipid metabolism.

Objective:To explore influence of walking training on pulmonary function and blood glucose in aged pa‐tients with type 2 diabetes mellitus (T2DM) .Methods :A total of 163 T2DM patients treated in our department were selected .According to number table ,they were randomly divided into routine group (n= 80 ,performed DM diet and routine insulin therapy strictly ,and lived as usual) and intensive group (n=83 ,received intensive walking train‐ing based on strict DM diet and routine insulin therapy ,namely walking ≥10000 steps/d for eight weeks) .Pulmona‐ry function and blood glucose indexes were compared between two groups before and after intervention .Results:Compared with before treatment ,after treatment ,there was significant rise in pulmonary function and significant reductions in blood glucose indexes in intensive group , P<0.01 all;compared with routine group after treatment , there were significant rise in vital volume [(2622.0 ± 323.3)ml vs .(2987.0 ± 405.6)ml] ,forced expiratory volume in one second [(2003.0 ± 279.7)ml比(2392.0 ± 323.6)ml]and vital volume/body weight [(44.0 ± 4.2) ml/kg vs . (47.0 ± 4.7) ml/kg] ,and significant reductions in levels of glycosylated hemoglobin A1c [(5.63 ± 2.63)mmol/L vs . (3.82 ± 1.41) mmol/L] ,fasting blood glucose [(7.90 ± 1.41) mmol/L vs .(5.28 ± 1.18) mmol/L]and 2h postpran‐dial blood glucose [(10.48 ± 1.43) mmol/L vs .(8.09 ± 1.68) mmol/L]in intensive group , P<0.01 all .Compared with before treatment ,there were no significant changes in above indexes in routine group after treatment , P>0.05 .Conclusion:As a low intensity aerobic exercise ,walking training can effectively improve pulmonary function , glucose metabolism in aged patients with type 2 diabetes mellitus ,it is help to improve quality of life .

BackgroundWith the number of diabetics increases,people pay more attention to the diabetic keratopathy.The major mechanism leading to diabetic keratopathy is diabetic corneal neuropathy.So it is significant to observe pathologic mechanism of diabetic corneal neuropathy. Objective To investigate the protective effects of edaravone( a free radical scavenger) on corneal nerve of rats with experimental diabetic corneal neuropathy,then explain the effects of oxidative stress in the pathologic mechanism of diabetic corneal neuropathy. Methods Seventy Sprague-Daxley male rats were taken as experimental subjects and 20 of them were used as normal control group.The remaining 50 were induced to be diabetic mellitus by a single intraperitoneal injection of streptozotocin and divided into 2 groups randomly:edaravone treated group and diabetic control group.In the edaravone treated group,edaravone(0.2 g/L) eye drops were used 3 times a day until the animal was killed.Five rats in each group were sacrificed at 6,8,10 and 12 weeks respectively.Then the corneal sensation,number of corneal nerve fibers,morphology,content of malondialdehyde(MDA) and activity of superoxide dismutase(SOD) in the corneal tissue were detected.ResultsIn the diabetic control group,the corneal sensation and the number of corneal nerve fibers were decreased,the density of neural network for cluster was sparse,the nerve activity was decreased,the content of MDA in the corneal tissue was significantly increased,the activity of SOD in the corneal tissue was significantly decreased (P＜0.01 ).Accompany with the course of disease,the above change was obvious day after day.Compared with the diabetic control group,the corneal sensation and the morphological abnormalities in corneal nerve of edaravone group were improved significantly which had the partial branches to the 12th week,the content of MDA in the corneal tissue was significantly decreased,the activity of SOD in the corneal tissue was significantly increased (P＜0.01).Conclusions Edaravone can lower diabetic corneal nerve of rats with experimental diabetic corneal neuropathyinjury,Oxidative stress may be a critical pathologic mechanism of diabetic corneal neuropathy.

As an important part of the health service security system,military medical equipment has important strategic position in wartime.Under the condition of high technology,the modern warfare has also laid higher requirement to the reliability guarantee of medical equipment.With the modernization development of military equipment,traditional medical equipment maintenance management mode does not meet the requirements of modern military medical equipment and its maintenance.Therefore,based on the reliability guarantee theory,the "Reliability Centered Maintenance(RCM)" management mode for military medical equipment is studied in order to achieve the purpose of improving the equipment reliability and the integrity of combat readiness.

Objective To investigate the expression pattern of Th1 and Th2 in patients with pulmonary tuberculosis including sputum positive pulmonary tuberculosis and coinfection with human immunodeficiency viruses(HIV), and its relation to disease severity.Methods The expression pattern of CD4+T lymphocytes, Th1 and Th2 cells in peripheral blood samples from sputum positive pulmonary tuberculosis patients, coinfection with HIV patients and healthy controls were studied by flow cytometry.Results The expression of CD4+ T lymphocytes and Th1 cells in sputum positive pulmonary tuberculosis patients were significantly lower than that in healthy group(31.22?9.80)%,(9.78?3.09)% vs (43.77?10.78)%,(25.26?4.73)%; The expression of Th2 cells in culture positive pulmonary tuberculosis patients was significantly higher than that in healthy group(18.65?3.49)% vs(10.15?2.60)%; The Th2 level in severe pulmonary tuberculosis patients was significantly higher than that in the medium and mild patients(21.70?2.67)% vs (14.87?1.66)% ,(17.48?2.06)%, while the CD4+T lymphocytes and Th1 levels were significantly lower; The CD4+ T lymphocytes and Th2 cells levels in pulmonary tuberculosis patients coinfection with HIV were lower than that in tuberculosis patients without HIV coinfection(21.88?3.71)%,(8.79?2.28)% vs (31.22?9.80)%,(18.65?3.49)%.Compared with the healthy group, Th1 cells level in pulmonary tuberculosis patients with or without coinfection of HIV were lower(25.21?4.73)% vs (9.39?2.65)%,(9.78?3.09)%.Conclusion Patients with sputum positive pulmonary tuberculosis showed lower expression of Th1 and higher expression of Th2,This profile was correlated with disease severity.Patients with pulmonary tuberculosis and HIV coinfection showed both of lower expression of Th1 without enhancement of the type 2 response.

Objeetive To explore the value of antisense imaging of 99Tcm-labeled ASON targeting mouse double minute 2(MDM2) mRNA for the diagnosis of human prostate cancer.Methods The ASON targeting MDM2 mRNA and the mismatched oligonucleotide (ASONM) were synthesized and radiolabeled with 99Tcm using the bifunctional chelator HYNIC.The labeling efficiency and radiochemical purity were investigated.Animal models of nude mice bearing human prostate cancer LNCaP were established and divided into 3 groups with 10 mice in each group.99Tcm-HYNIC-ASON,99Tcm-HYNIC-ASONM (study groups) and 99TcmO4-(control group) were injected at the dose of 7.4 MBq through the tail vein,respectively.Tumor imaging was acquired with SPECT and the tumor-to-muscle (T/M) ratio was measured.The data was compared by one-way analysis of variance.Results The labeling efficiencies of ASON and ASONM were (65.15± 2.05) ％ and (64.93±2.18) ％,respectively.Their radiochemical purity was greater than 90％.At 1,4 and 10 h post injection,the T/M ratios of 99Tcm-HYNIC-ASON group were 3.217±0.125,3.749± 0.201 and 4.028±0.186,and those of 99Tcm-HYNIC-ASONM group were 1.579t0.128,1.715±1.140 and 1.683±0.139,and control group 2.146±0.132,1.847±0.124,1.528±0.152,respectively.The T/M ratios in control group and 99Tcm-HYNIC-ASONM group were significantly lower than those in 99Tcm-HYNICASON group at 1,4 and 10 h,respectively (F=213.37-235.41,t=3.527-4.738; all P＜0.01).The T/M ratios of 99Tcm-HYNIC-ASONM group and control group were not significantly different at 1,4 and 10 h (t=2.154,2.287 and 2.236,all P＞0.05).Conclusion The antisense probe of MDM2 can accumulate specifically in prostate cancer tissue in animal models,which might be useful as a non-invasive genetic tool for the early diagnosis of prostate cancer.

Objective To investigate the effect of mouse double minute 2 (MDM2) mRNA ASON and mismatched oligonucleotide (ASONM) radiolabeled with 99Tcm on target gene expression in LNCaP cells.Methods The ASON and ASONM targeted to MDM2 mRNA were synthesized and radiolabeled by 99Tcm with the bifunctional chelator of HYNIC.The labeling efficiency,radiochemical purity,stability and molecular hybridization activity were investigated.The different concentrations of 99Tcm-HYNIC-ASON (0,100,500 nmol/L) and 99Tcm-HYNIC-ASONM (500 nmol/L) coated with lipofectamin 2000 were incubated with prostate cancer cells for 24 h,then RT-PCR and Western blot were carried out to assay the MDM2,p53 mRNA and the corresponding protein level.The variables of RT-PCR and Western blot were analyzed using one-way analysis of variance and q test.Results The labeling efficiency of ASON and ASONM were (65.15± 2.05)％ (n=5) and (64.93±2.18)％ (n=5),respectively.The radiochemical purity were both more than 90％.99Tcm-HYNIC-ASON had a good stability and could hybridize to the sense oligonucleotide (SON).The contents of MDM2 mRNA in 0,100,500 nmol/L 99Tcm-HYNIC-ASON and 500 nmol/L 99Tcm-HYNIC-ASONM groups were 0.458±0.035,0.250±0.026,0.174±0.032,0.463±0.033,respectively,and there were significant differences between each 2 groups except between 0 nmol/L 99Tcm-HYNIC-ASON and 500 nmol/L 99Tcm-HYNIC-ASONM groups (F=33.69,q =24.32-91.45,all P＜0.01).The average density of MDM2 protein in the 4 groups were 90.712±3.042,71.218±2.915,32.775±3.062,88.121±2.710,respectively (F=235.93,q=6.43-19.14,all P＜0.01; except 0 nmol/L99Tcm-HYNIC-ASON and 500 nmol/L 99Tcm-HYNIC-ASONM).The contents of p53 mRNA in the 4 groups were 0.185±0.046,0.203±0.040,0.213±0.027,0.163±0.049,respectively(F =2.18,P＞ 0.05).The average density of p53 protein was 33.865 ± 2.213,70.445±2.180,99.025±3.012,38.351±3.271,respectively (F=53.98,q =3.32-6.74,all P＜0.01 ; except 0 nmol/L 99Tcm-HYNIC-ASON and 500 nmol/L 99Tcm-HYNIC-ASONM).Conclusions The MDM2 antisense probe can accumulate in the prostate cancer cells,and specially hybridize to the MDM2 mRNA and inhibit target gene expression.This novel molecular probe has a promising potential for the diagnosis of prostate cancer at gene level.

BACKGROUND:Isoflurane cannot only induce a wide range of large neuronal apoptosis, but also inhibit hippocampal neurogenesis in neonatal rats, thereby resulting in hippocampus-dependent learning and memory defects.
OBJECTIVE:To investigate the isoflurane effect on proliferation and differentiation of the hippocampal neural stem cels.
METHODS:Twenty-six Sprague-Dawley rats were randomly divided into air group and isoflurane group (n=13 per group). Rats in the isoflurane group were subjected to 2.5% isoflurane inhalation for 3 minutes folowed by 1.5% isoflurane inhalation for 4 hours. Rats in the air group only breathed in air. After the intervention, blood glucose and arterial blood gas changes were detected in the two groups. Additionaly, rats in the two groups were given intraperitoneal injection of 5-bromodeoxyuridine before and after intervention. At 24 hours after the last injection of 5-bromodeoxyuridine, brain tissues were taken to make frozen sections for immunofluorescence staining.
RESULTS AND CONCLUSION:There were no significant difference in pH, PaO2, PaCO2, HCO3, BE and SaO2 levels between the two groups (P> 0.05). Compared with the air group, the number of BrdU+ cels was significantly less in the isoflurane group (P < 0.05), while the number of NeuroD+/BrdU+ cels was significantly higher in the isoflurane group (P < 0.05). The incidence of adverse reactions was 23% in the isoflurane group, which was significantly higher than that in the air group (7.7%;P < 0.05). These findings indicate that isoflurane can inhibit the proliferation of neural stem cels in the hippocampal dentate gyrus, and promote their differentiation into neurons.

Objective To investigate the regularity and significance of serum interleukin-6(IL-6) levels in the patients with sepsis.Methods Forty sepsis patients with APACHEⅡ scores over 12 and 30 normal controls were involved in the study.The IL-6 levels in serum were detected with radio-immunity analysis assay and the correlations among the IL-6 levels,the prognosis and APACHEⅡ score were investigated.Results In the patients with sepsis,the average level of IL-6 in serum was(152.02?55.77) pg/ml,which was significantly higher than that of the control group [(85.79?30.96) pg/ml](P

Starting with the concepts of basic dataset and basic dataset of nursing management information of hospital of Tradtional Chinese Medicine (TCM),the paper introduces the research purpose,construction principle,construction method,construction contents and system framework of the basic dataset standard system of nursing management information of hospital of TCM,in order to lay a foundation for achieving nursing management data resource sharing and exchange of hospital of TCM.