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 .

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.

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.

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.

BACKGROUND:Experiments have shown that the col agen substrate has the capability of stimulating cartilage generation, but the stimulating role of different types of col agen substrates remains controversial. OBJECTIVE:To investigate the effect of type I and type II col agen on the biological characteristics of human chondrocytes cultured in vitro. METHODS:Human chondrocytes at passage were cultured onto the ordinary culture plates (ordinary plate), type I col agen-coated culture plates (type I plate), and type II col agen-coated culture plates (type II plate). cellgrowth curves were determined by MTT method after cells were cultured for 10 days. By ELISA, PCR, and 1,9-dimethyl methyleneblue technology, type I and type II col agen and glycosaminoglycan contents were quantitatively detected in cartilage cells 28 days after culture. RESULTS AND CONCLUSION:The number of cartilage cells was the highest in type II plate, which was twice of that in type I plate and five times of that in ordinary plate. Cartilage cells in type II plate secreted the least amount of type I col agen, which showed significant differences compared with the ordinary plate (P<0.01) and had no statistical y significant difference with type I plate (P>0.01). Cartilage cells in type II plate secreted the most amount of type II col agen and glycosaminoglycan, showing significant differences compared with the other two plates (P<0.01). The cartilage cells cultured in col agen plates are better than that cultured in ordinary culture plate, type II col agen culture plate is better than type I col agen culture plate in maintaining cellshape, extending the dedifferentiation pattern, and promoting celldifferentiation.

Objective To explore the expression of cyclooxygenase-1 (COX-1) in cervical carcinoma and its significance. Methods The pathological specimens were collected from 62 female patients, who were admitted to 307 Hospital of PLA from Jan. 1999 to Mar. 2005, including 31 cases of cervical carcinomas, 15 cases of cervical intraepithelial neoplasia and 16 cases of normal cervix. Surgery or biopsy was performed. Expression of COX-1 was detected by immunohistochemistry, and the relationship between COX-1 and clinicopathological feature was analyzed. Results The major sites of COX-1 expression were localized in cytoplasm, and next in cell membrane. Strongly positive expression of COX-1 was observed in cervical carcinomas, and weakly positive expression of COX-1 in cervical intra-epithelial neoplasia, with positive rates of 81% and 13%, respectively. There was no COX-1 expression in normal cervix. A significant difference was observed among these specimens. No obvious correlation was found between COX-1 expression and patient's age, tumor differentiation degree snd clinical stages. Conclusion Expression of COX-1 may serve as an auxiliary parameter for diagnosis, therapeutic scheme option, and prognosis of patients with cervical carcinoma.

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

Objective To investigate the effect of valsartan,an angiotensinⅡtype 1 receptor (AT1 R)blocker,on radiosensitivity,invasive potential and proliferation activity of nasopharyngeal carcinoma cells(CNE-2)in vitro. Methods Radiosensitization of valsartan on CNE-2 cells in vitro was investigated by colony forming assay.Effect of ATl R blocker combined with radiation on invasive potential of CNE-2 cells was evaluated using 24-well Matrigel invasion chambers(Transwell).Apoptosis-inducing effect of valsartan combined with radiation on apoptosis of CNE-2 was identified by flowcytometry(FCM). Resuits When valsartan was given at 10-9.10-8 and 10-7 mol/L combined with radiation,sensitivity enhancement ratios (SER)were 1.10,1.20 and 1.36.and the invasive inhibition rates were 8.11%,16.49%and 16.77%,respectively.The SER of valsartan on CNE-2 distinctly increased when the exposure time was increased.After 24 h exposure to 10-8 mol/L valsartan combined witIl radiation.the apoptosis rate was 1.89%±0.09%,which was higher than 1.62%±0.06%in radiation alone group(t=4.79.P＜0.05). Conclusions AT1 R blocker valsartan combined with radiation can significantly inhibit the proliferation activity of nasophar,cngeal carcinoma cells in vitro in a dose- and time-dependent manner.Valsartan combined with radiation can potently inhibit the invasive potential of CNE-2.which may be involved in the mechanism of valsartan treatment in vivo.

OBJECTIVE:To screen the optimal technology of dog testes and penis processed with talcum powder,and to pro-vide reference for the formulation of quality control standard. METHODS:Taking the comprehensive score of crushing rate and the content of alcohol soluble extract as evaluation indexes,L9(34) orthogonal test was used to optimize processing temperature,pro-cessing time and the amount of talcum powder;validation test was conducted. According to related method in Chinese Pharmaco-poeia(2015 edition),the contents of moisture,ash,nitrogen and alcohol soluble extract were determined,and automatic amino ac-id analyzer was used to determine the content of amino acids in samples. RESULTS:The optimal processing technology were that 100 kg dog testes and penis should add into 40 kg talcum powder;the processing temperature was 350-380 ℃;processing lasted for 4 min. Results of verification test showed that the average crushing rate and the content of alcohol soluble extract were 80.2%(RSD=0.95%,n=3)and 15.7%(RSD=2.30%,n=3). In processed dog testes and penis,the contents of moisture,ash,nitro-gen and alcohol soluble extract were 5.7%,18.7%,8.3%,15.7%,respectively. Besides,there still were 16 kinds of amino ac-ids,and their total content was 42.44%. CONCLUSIONS:The optimized talcum powder processing technology of dog testes and penis is stable and practical. The content of moisture,ash,nitrogen,alcohol soluble extract and amino acid can be used as impor-tant quality standard control indexes for processed dog testes and penis.