Aim To explore the role of nicotinamide adenine dinucleotide phosphate ( NADPH ) oxidase in high glucose-induced injury in H9c2 cardiac cells. Methods H9 c2 cardiac cells were exposed to differ-ent concentrations of high glucose(5. 5 mmol·L-1 ,11 mmol · L-1 ,22 mmol · L-1 ,33 mmol · L-1 ,44 mmol ·L-1 , 55 mmol · L-1 ) for 24 h and different time pints of high glucose(0 h,12 h,24 h,36 h,48 h,72 h) . Cell viability was measured by MTT colorimetry, the protein expression of Bcl-2 , Bax and NADPH oxi-dase submits such as p22 phox , p47 phox and p67 phox were determined by western blotting. Results H9c2 cardi-ac cells exposure to high glucose for 24 h showed on decrease in cell survival and the Bcl-2 expression while an increase in the Bax expression ( P<0. 05 ) . Moreo-ver, high glucose could markedly up-regulate the activ-ity of NADPH oxidase characterized by the enhanced expression of p22 phox , p47 phox and p67 phox ( P<0. 05 ) . Conclusion Activating NADPH oxidase may play an important role in high glucose-induced injury in H9 c2 cells.

For screening the potential drugs as anti-liver fibrosis candidates, we established a high- throughput drug screening cell model based on COL1A1 promoter. The activity of COL1A1 promoter and luciferase reporter gene can be elevated by TGF-β1, and inhibited by candidate drugs. We constructed a recombined plasmid with COL1A1 promoter and luciferase reporter gene pGL4.17, the activity of COL1A1 promoter was reflected by fluorescence intensity. COL1A1 promoter activity was detected by Dual-Luciferase Reporter Assay System, it came that the relative luciferase activity of COL1A1 promoter was 15.98 times higher than that of control group induced by TGF-β1, showing the recombined plasmid could be used in cell model. The recombined plasmid was transfected into human hepatic stellate cells LX2, detected the effect of potential drugs, and obtained a stable expression system through stable transfection and monoclonal cell culture. A sample which could reduce COL1A1 promoter activity signally by our cell model, decreased collagen I mRNA and protein expression detected by real-time RT-PCR and Western blotting. It indicates this novel cell model can be used in high-throughput drug screening of potential anti-liver fibrosis drugs.
Collagen Type I ; genetics ; Drug Evaluation, Preclinical ; methods ; Genes, Reporter ; Hepatic Stellate Cells ; High-Throughput Screening Assays ; Humans ; Liver Cirrhosis ; drug therapy ; Luciferases ; Plasmids ; Promoter Regions, Genetic ; RNA, Messenger ; Transfection ; Transforming Growth Factor beta1 ; pharmacology

Bone Neoplasms ; Female ; Humans ; Middle Aged ; Nasal Septum ; Nose Neoplasms ; Osteoma

ObjectiveTo evaluation application of MSCTA technology in pre-and pro-liver transplantation.MethodsThirty - two patients with liver transplantation were performed with multi-phase CT scanning (MSCT).In hepatic artery and portal phase vascular 3D imaging reconstruction methods include multiple planar reconstruction ( MPR),maximum intensity projection ( MIP),volume rendering (VR).MIP images were measured with these parameters (CA,LGA,CHA,PHA,PV,SV,SMV in diameter).ResultsHepatic arterial vascular imaging scan can show clearly within the scope of the abdominal aorta,gastroduodenal artery,hepatic artery,the left hepatic,celiac trunk,and its branches.Portal vascular imaging clearly showed that the portal vein system.MIP can use accurate measurement of vascular diameter abdominal aorta and portal,superior mesenteric vein,and splenic vein diameter.Control group with liver cirrhosis and liver cancer artery diameter had no significant difference in patients with portal hypertension and the portal vein [ ( 16.7 ± 2.4 ) mm ],mesenteric vein [ ( 10.8 ± 2.1 ) mm ],and splenic vein diameter [(13.1±1.9)mm] compared to the control group[(13.8 ±1.6)mm,(8.2 ±1.7)mm,(8.9±1.1)mm ],the difference was statistically significant ( P ＜ 0.05 ). Conclusions MSCTA can show the liver artery and portal vein system of the non-invasive method of checking,Joint MIP and VR application can provide more clinical liver transplant before the hepatic artery and portal vein of information,liver transplantation arteriovenous anastomoses provides for the monitoring of vascular diameter.

Objective: To measure serum testosterone level, plasma levels of interleukin-18 (IL-18) and IL-10 and explore their correlation in patients with different types of coronary heart disease (CHD), and their possible role in occurrence and development of CHD. Methods: A total of 96 male CHD patients were divided into acute myocardial infarction (AMI) group (n=35), unstable angina pectoris (UAP) group (n=32) and stable angina pectoris (SAP) group (n=29). Another 30 patients who were excluded for CHD by coronary angiography were enrolled as non CHD control group. Enzyme linked immunosorbent assay (ELISA) was used to measure serum testosterone level and plasma levels of IL-18 and IL-10 in all groups. Results: Compared with non CHD control group, there were significant decreases in serum testosterone level [(13.46±1.99) mmol/L vs. (6.89±1.35) mmol/L vs. (5.02±1.87) mmol/L, t=1.917～2.365, P<0.05 both] in UAP group and AMI group, and that of AMI group was significantly lower than that of UAP group, t=1.034, P<0.05; there were significant increases in IL-18 levels [(146.72±79.36) pg/ml vs. (209.32±80.49) pg/ml vs. (316.78±75.63) pg/ml vs. (457.78±83.21) pg/ml, t=2.016～3.167,P<0.05 all] in SAP group, UAP group and AMI group, and those of UAP group and AMI group were significantly higher than that of SAP group, t=2.173, 2.596, P<0.05; there were significant increases in IL-10 levels [(48.46±18.27) pg/ml vs. (116.45±42.76) pg/ml vs. (85.64±27.33) pg/ml vs. (70.26±18.55) pg/ml, t=2.997～2.018, P<0.05 all] in SAP group, UAP group and AMI group, and those of AMI group and UAP group were significantly lower than that of SAP group (t=2.034, 2.291, P<0.05 both). Pearson linear regression analysis indicated that serum testosterone level was negatively correlated with levels of IL-10 (r=-0.678, P<0.01) and IL-18 (r=-0.579, P<0.01) in CHD group. Conclusion: There are significant changes in serum testosterone level and plasma IL-18, IL-10 levels, and testosterone level is significantly negatively related with IL-18, IL-10 levels, and they can be regard as new indexes assessing coronary atherosclerotic lesion.

Objective To observe the changes of serum sphingosine-1-phosphate (S1P) level in asthmatic patients with different severity of bronchial asthma, and to explore the evaluation value of S1P on the severity of asthma.Methods A prospective observational study was conducted. Fifty-two patients with asthma admitted to Department of Respiratory Medicine of the First Affiliated Hospital of Jiamusi University from November 2015 to January 2017 were enrolled. According to the severity of the disease, the patients were divided into mild, moderate and severe groups. In the same period, 25 healthy subjects were served as healthy control group. All the subjects got the peripheral venous blood collection in the morning fasting, the level of serum S1P was determined by enzyme linked immunosorbent assay (ELISA), the peripheral blood eosinophil (EOS) was counted, and the pulmonary function test was performed. The correlation among the parameters was analyzed by Pearson correlation analysis. Receiver operating characteristic curve (ROC) was plotted, and the value of serum S1P on evaluating the severity of asthma was analyzed.Results Fifty-two asthma patients were enrolled, including 17 patients of the mild, 19 of the moderate, and 16 of the severe. Compared with the healthy control group, serum S1P level and peripheral blood EOS in different degree asthma groups were significantly increased, and forced expiratory volume in 1 second (FEV1) was decreased significantly; and with asthma exacerbations, serum S1P levels and peripheral blood EOS were gradually increased [mild, moderate and severe S1P (nmol/L) were 1537.0±120.3, 1980.7±149.5, 2202.2±117.2 (F= 274.624, P= 0.001); EOS (×109/L) were 0.13±0.06, 0.20±0.07, 0.37±0.14 , respectively (F= 44.093,P = 0.001)], and FEV1 was decreased gradually [mild, moderate and severe were 0.89±0.05, 0.63±0.06, 0.42±0.10, respectively (F= 159.756,P = 0.001)]. Correlation analysis showed that there were significant positive correlations between serum S1P level and peripheral blood EOS in patients with mild, moderate and severe asthma (r value was 0.696, 0.746,0.508, allP < 0.05), and negatively correlations with FEV1 were found (r value was -0.761, -0.655, -0.815, all P < 0.01). There was no significant correlation between serum S1P level and EOS, FEV1 in healthy control group (r value was 0.324 and -0.048, bothP> 0.05). ROC curve analysis showed that the area under curve (AUC) of serum S1P for assessing mild, moderate and severe asthma was 0.948, 1.000, 1.000, respectively; when the cut-off of S1P was 1181.8, 1534.2, 1708.6 nmol/L, the sensitivity was 88.2%, 100%, 100%, and the specificity was 88.0%, 100% and 100%, respectively.Conclusions During asthma attack, the serum S1P level was gradually increased with the exacerbation of the disease. Serum S1P level has significant evaluative effect on the severity of asthma.

Currently, competition in the market of health service of our country is very fierce. In this circumstance, realization of military hospital's development aim-focuse on people, leading position in technology-to provide better service armyman is depend on economic foundation. To increase economic benefit of hospital, it is necessary to intensify management of cost accounting first, in which cost accounting of clinical department is of especial importance. In this article, we probe into the significance, content, method and problems of cost accounting in clinical department of military hospital.

Objective:To determine if suitable electric pulses-mediated DNA and DNA and bupivacaine complexes delivery technologies could enhance effects of botulinum neurotoxin serotype A (BoNT/A) DNA vaccines in mouse model. Methods:Vaccination of mice i.m. with plasmid DNA replicon vaccine pSCARSHc and conventional plasmid DNA vaccine pcDNASHc following electric pulses and with DNA and bupivacaine complexes. AHc-specific group antibody ELISA titers and lymphocyte proliferative responses of mice were detected and IgG1 and IgG2a isotype profiles were assayed. Results:Immune effects of DNA vaccines were enhanced following electric pulses and bupivacaine delivery. Effects of DNA vaccines following electric pulses were better than that of DNA vaccines formulated with bupivacaine,and the combined delivery technology of electric pulses and bupivacaine induced the highest level of specific antibodies and lymphocyte proliferative responses. Plasmid DNA replicon vaccine pSCARSHc induced relatively higher AHc-specific antibodies and lymphocyte proliferative responses in immunized Balb/c mice than conventional plasmid DNA vaccine pcDNASHc in these DNA delivery technologies. And vaccine pSCARSHc induced Th2/Th1-type immune responses with a general bias to Th2-type,and vaccine pcDNASHc induced Th2-type immune responses. Conclusion:Suitable electric pulses-mediated DNA and DNA and bupivacaine complexes delivery technologies could enhance effects of BoNT/A DNA vaccines in mouse model. Therefore,the methods described here potentially provide suitable strategies in developing an efficacious vaccine against botulinum neurotoxin serotype A.

OBJECTIVE: To establish a multiplex STR genotyping method for autosomal STR and Y-STR loci in forensic biological practice. METHODS: Widely used autosomal STR loci and Y-STR loci were selected. A set of PCR primers was designed, and a 5-dye fluorescent labeled STR multiplex PCR reagent kit was developed. RESULTS: A kit was developed which can simultaneously detect 15 autosomal STR loci, 10 Y-STR loci, and an Amelogenin. CONCLUSION The 15 autosomal STR plus 10 Y-STR kit in combination with capillary electrophoresis method was used to STR genotyping with accurate and reliable results. The new one-step testing kit can potentially be widely used in forensic cases and DNA databank in the future.
Amelogenin ; Chromosomes, Human, Y/genetics* ; DNA Primers ; Databases, Nucleic Acid ; Forensic Genetics/methods* ; Genotype ; Genotyping Techniques/instrumentation* ; Humans ; Indicators and Reagents ; Microsatellite Repeats ; Multiplex Polymerase Chain Reaction

A completely synthetic gene encoding the He domain of Clostridium botulinum neurotoxin serotype A (AHc, 1287 bp, 429 aa, -50 kD) was constructed with oligonucleotides. After expressed in Escherichia coli, soluble product AHc was gained and verified by SDS-PAGE and Western blot analysis. The expressive level of recombinant AHc in E. coli was very high (36%-53% of soluble total proteins) and the purified yield was more than 30 mg/L by one-step purification. Then, the purified AHc was used to vaccinate Balb/c mice, which developed a strong and specific immune response as expected following administration of AHe protein via the subcutaneous route. Results from BoNT/A neutralization assay showed that the serum from mice vaccinated with AHc contained high titer protective antibody. These results showed that the soluble, stable and high-levelly expressive AHc not only could be produced by the prokaryotic expression system built in our lab, but also owned strong immunogenicity to prepare antitoxin for treatment and as sub-unit candidate vaccine for prophylaxis against botulinum toxin serotype A.
Animals ; Antibodies, Bacterial ; blood ; Bacterial Vaccines ; genetics ; immunology ; Botulinum Toxins, Type A ; biosynthesis ; genetics ; immunology ; Botulism ; immunology ; prevention & control ; Clostridium botulinum type A ; genetics ; immunology ; Escherichia coli ; genetics ; metabolism ; Female ; Lymphocyte Activation ; Mice ; Mice, Inbred BALB C ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; T-Lymphocytes ; immunology ; Vaccines, DNA ; genetics ; immunology