Objective To investigate the changes in iNOS expression in brain injury induced by hepatic ischemia-reperfusion (IR) in rats.Methods Thirty-two male Wistar rats weighing 240-280 g were randomly divided into 2 groups: sham operation group (group S,n = 8) and group IB( n = 24).The hepatic IR was induced by clamping the hepatic artery and portal vein according te the method described by LONG et al.In group IR the rats were killed at 3,6 and 24 h of reperfusion after 40 min hepatic ischemia (8 rats at each time point).The rats in group S were also killed.The brains were removed for determination of NO content (by nitrate reductase assay),SOD activity (by xanthine oxidase method),MDA content(by colorimetric method),nitrotyrosine (NT) expression (by Western blot),and iNOS mRNA expression (by RT-PCR).Results Compared with group S,cerebral NO and MDA content were significantly increased at 6 and 24 h of reperfusion,expression of cerebral NT and iNOS mRNA up-regulated and SOD activity decreased at 6 h of reperfusion in group IR (P < 0.05 or 0.01).Cerebral NO and MDA content were significantly higher and SOD activity lower at 6 and 24 h of reperfusion than at 3 h of reperfusion in group IR (P < 0.05).Conclusion The expression of iNOS in brain tissues is up-regulated after hepatic IR and it produces a great amount of NO inducing brain injury through peroxynitrite (ONOO-).

Objective To evaluate the clinical diagnostic value of nucleic acid amplification (TB- RNA),bacteriophage-based assay,3D culture and smear on the detection of Mycobacteria tuberculosis.Methods 291 clinical sample including 110 sputum,54 thoracic fluid,37 throat swab,31 bronchial fluid,13 cerebrospinal fluid,12 urine,8 lymph fluid and 20 others (pericardial effusion,feces, blood and abdominal fluid) and gynecological specimen (including 6 leucorrhoea and menstrual blood) were analyzed by these four methods.Results Among the 291 clinical samples,the positive rate of mycobacteria tuberculosis for TB-RNA,bacteriophage-based assay,3D culture and smear were 37.1%,28.9%,27.5% and 10.3%.The sensitivity and specificity of the TB-RNA,bacteriophage-based assay,3D culture and smear were 54.3% & 100%,41.7% & 88.9%,31.7% & 93.5% and 14.6% & 98.9%,respectively.Conclusions TB-RNA is an effective clinical diagnostic method for Mycobacteria tuberculosis.Although the sensitivity of smear is poorer than others,it is a universal testing method in clinical laboratory due to low cost.The positive rate of mycobacteria tuberculosis for 3D culture is lower than that of bacteriophage-based assay and TB-RNA.Although the time to result for 3D culture might last for few weeks,the isolates can be used for drug resistance screening and bacterial identification.

Objective To study the application effect of day-parting appointment for elderly hypertensive contracted outpatients in community. Methods In May 2018 two groups (experiment and control) of 103 elderly hypertensive contracted outpatients, aged between 60 and 80 and looked after by the team of family doctors, who had been diagnosed with hypertension and with medication for at least one year were set up.The experimental group used self-made community hypertension visiting card for appointments, and the control group used the original way of treatment.Six months later, comparison was made in blood pressure control, the number of outpatients, the time consumed and the satisfaction between the two groups. Results It was found in comparison that the blood pressure standard-reaching rate of the experimental group was better than that of the control group (P < 0.05);outpatients′ visits and time consumed were less than those of the control group, the time used in the clinic room was longer than that in the control group; the differences in these aspects between the two groups were significant (P < 0.05);the satisfaction of the overall outpatient perception, attitude of family doctors, treatment technology, visits control, dosage, cost and safety of medication in the experimental group were better than in the control group; there were significant differences in these aspects between the two groups (P < 0.05). Conclusion Community day-parting appointment proves to be more convenient, more time-saving and safer for outpatients, greatly improving the medical quality and satisfaction for community outpatients.

A strain was isolated from internal organ of died porcine about 8 weeks with purulent pneumonia,arthritis,pyogenic arthritis and endocarditis in April 2007.Objectives of the study are to confirm the genus of the strain,pathopoiesis,and drug sensitivity.The mainly study methods:the first,the strain was identified by the phenotype and the characteristics of the biochemistry,sequence 16S rDNA genes of the strain was analyzed by molecular biology technology,finally animal experiment and drug sensitivity testing were done.The results of the phenotype and the characteristics of the biochemistry showed that it is greatly similar to Actinomyces hyovaginalis,16S rRNA sequence analysis exhibited the homology achieved to 99.2% com-pared with group III strains of Actinomyces hyovaginalis,and the phyletic evolution analysis also indicated that it has mostly relationship with group III strains of Actinomyces hyovaginalis.Animal experiment dis-covered it has highly pathogenicity to Mus musculus albus;Drug sensitivity testing showed that it is hyper-sensitive to Erycin,Gentamicin and Amikacin.So,the result of the study confirmed that the strain is Actin-omyces hyovaginalis III with the pathogenicity.

Classical swine fever virus (CSFV), an enveloped positive-stranded RNA virus in the genus Pestivirus of the Flaviviridae family, is the causative agent of a highly contagious swine disease characterized by symptoms of hemorrhagic fever and immune depression, usually leading to substantial economic losses. The serological methods for detection of CSFV antibody such as ELISA are important means for the diagnosis of CSFV and immune surveillance. It is difficult to obtain CSFV antigen with high quality using traditional method because its titration titer is low in cell culture. CSFV has four structural protein named C, E0, El and E2. The E2 protein contains major antigenic determinants that are conserved between different CSFV strains and involved in neutralization by antibodies. So recombinant E2 protein can be developed as an alternative to the intact viral antigen. So far, CSFV E2 have not been expressed in E. coli with high level. Many factors, such as the secondary structure, the stability of 5' and 3' terminus of gene, the location of SD sequence and the bias of codes, are involved in the expressing level of foreign gene in E. coli . In this study, two sites of the E2 gene sequence were confirmed to be detrimental to its expression efficiency in E. coli through the computer-aided analysis. So they were mutated using recombinant PCR without changing the amino acids sequence of CSFV E2 gene. A plasmid was constructed by inserting the mutated E2 gene into the prokaryotic expression vector pET-28a(+) and named pETE2. The E. coli competent host BL21 (DE3)lysS transformed with pETE2 could express the E2 gene at high level, amounting to 28% of the total protein of the induced recombinant bacteria at the presence of IPTG. Except the hydrophobic transmembrane domain at C terminus, the recombinant E2 protein includes the total aa sequence. So it contains all the potential linear antigen epitopes of E2 protein because hydrophobic aa region can not form epitope. The recombinant E2 protein was CSFV-specific as proved by Western blotting and indirect ELISA. The rabbits immunized with the recombinant E2 can be protect from the challenge of hog cholera lapinized virus. This is the first report that E2 gene is expressed with high level expression in E. coli. In conclusion, it is an effective measure that mutate the CSFV E2 gene to increase its expression level in E. coli. The recombinant CSFV E2 protein possess fine immunonicity and can be used the antigen for the detection of CSFV antibody.
Antigens, Viral ; genetics ; immunology ; metabolism ; Blotting, Western ; Enzyme-Linked Immunosorbent Assay ; Escherichia coli ; genetics ; metabolism ; Mutagenesis, Site-Directed ; methods ; Polymerase Chain Reaction ; Recombinant Proteins ; genetics ; immunology ; metabolism ; Viral Envelope Proteins ; genetics ; immunology ; metabolism

Factor VII Deficiency ; diagnosis ; genetics ; Female ; Humans ; Middle Aged ; Pedigree ; Phenotype

OBJECTIVETo construct luxS mutant aften luxS gene of Streptococcus mutans (S. mutans) was knocked out, and examine their ability of biofilm formation.

METHODSA recombinant plasmid containing the flanking fragment of luxS of S. mutans was transformed into S. mutans UA159, and selected by brain heart infusion (BHI) agar medium with kanamicin. The luxS mutant further confirmed via polymerase chain reaction (PCR) and the autoinducer-2 (AI-2) bioluminescence assay of Vibrio harveyi (V. harveyi), and ability of luxS mutant and S. mutans UA159 biofilm formation was examined in different phases, in BHI medium with 1% sucrose and 1% glycose by scanning electron microscopy (SEM).

RESULTSLuxS-deficient S. mutans strains were successfully constructed. Compared with S. mutans UA159, the luxS mutant maintained in BHI medium containing 1% sucrose displayed an apparent defect in biofilm formation, while they showed no significant deviation in BHI medium containing 1% glycose.

CONCLUSIONluxS gene in S.mutans can play a role in dental plaque biofilm formation, and the luxS gene is possible to regulate sucrose-dependent biofilm formation.

Bacterial Proteins ; Biofilms ; Carbon-Sulfur Lyases ; Culture Media ; Dental Plaque ; Homoserine ; analogs & derivatives ; Humans ; Lactones ; Streptococcus mutans

OBJECTIVETo explore the thermal treatment schedule of leucite microcrystallization to reinforce dental glass ceramics.

METHODSAfter component analysis and selection, the raw material were treated by different temperature schedules. The products were analyzed by polaring microscope and X-ray diffractometer to determine the appropriate thermal treatment schedule.

RESULTSThe temperature of melting, nuclearing and crystalizing was 1,600 degrees C, 1,200 degrees C and 1,500 degrees C. Leucite microcrystals dispersed in the glass matrix evenly and the size of leucite particle was about 0.8 micro m.

CONCLUSIONLeucite can be microcrystalized according to an appropriate thermal treatment schedule.

Aluminum Silicates ; chemistry ; Crystallization ; Dental Porcelain ; chemistry ; Glass ; chemistry ; Hot Temperature

OBJECTIVETo compare the effect of Sodium Morrhuate on ECV-304 between its lipo- and normal preparation.

METHODSThe ECV-304 cell line was supplemented with Sodium Morrhuate and lipo-Sodium Morrhuate in order, and the result on morphology (microscope, Giemsa Staining and electron microscope), cell activity (MTT), and flow cytometer between the two preparation were compared.

RESULTSIn normal preparation group, cell's edema occurred. Chromatin was like catkins. Tumefaction and degeneration of mitochondrion and endoplasmic reticulum appeared. In lipo-Sodium Morrhuate group, the membrane was creased and processus appeared. Chromatin aggregates to the membrane of nucleus was like crescent, and then broken. The apoptotic body was formed. MTT changes showed that the curve of the normal preparation group was steep and the change time was short relatively, which cues the vital cells decreased sharply. The curve of lipo-Sodium Morrhuate group was gentle and the change time was long relatively, which cues the vital cells decreased slowly. The flow cytometer showed that typical apoptosis peak appeared.

CONCLUSIONThe normal preparation group shows an acute toxic effect on ECV-304 cell line, which result in a necrosis course, while lipo-Sodium Morrhuate shows a gradual releasing process, which may indicate a apoptosis course.

Animals ; Apoptosis ; Cell Line ; Humans ; Necrosis ; Sclerosing Solutions ; Sodium Morrhuate

Animals ; Asia ; epidemiology ; History, 20th Century ; Humans ; Influenza A Virus, H2N2 Subtype ; chemistry ; genetics ; isolation & purification ; Influenza, Human ; epidemiology ; history ; virology ; Mutation