Thermal Shift Assays was applied for high-throughput screening optimized buffer system used in the gel filtration chromatography for purification of recombinant TBEV serine protease. The optimization included kinds of salt and pH. It was optimized new buffer remarkably improved the quality of the purified TBEV serine protease by increasing its monomer from 75% to 99%. This result may improve future crystal studies of this protein.

Objective To investigate the source of CD147 expressing cells in joint and its corelation with rheumatoid arthritis (RA),and to study the expression of CD147 on different cells of peripheral blood and joint fluid of RA patients and normal health controls.Methods The expression rate and intensity of CD147 on lymphocyte (CD3 +T),monocyte (CD14 + Mo) and neutrophils (N) of 10 active RA patients before and after treatment and 10 normal health controls′ peripheral blood and synovial fluid were studied by bicolor immunofluorescence flow cytometry.Results ① Compared with normal control,the expression intensity of CD147 on CD3 + T and CD14 + Mo were higher (P

OBJECTIVE To analyze the nosocomial infections among dead cases with cancer and explore the relationship between death and nosocomial infections.METHODS Totally 309 dead cases with cancer were investigated and analyzed in 2004-2005.RESULTS From them 64 with 72 times accompanied by nosocomial infections(20.71%,23.30%),the rate was 5.85 times higher than the total nosocomial infection rate in the hospital.Of 64 cases with nosocomial infections,48(75%) were led to death directly by nosocomial infections.The average hospitalization time of these 64 dead cases with nosocomial infections was 63.94 days,which was 20.98 days longer than the whole average hospitalization time(29.21 days).Cases with tumors related to immune system were more often accompanied by nosocomial infections than with other tumors(P

Based on the concept of environmental factors of ICF, together with the practice of environmental assessment and reform in the recent years, this paper discussed the principle, specification, procedures and content of environmental assessment.

AIM: To study the causes of pyogenic granuloma after hydroxyapatite(HA) orbital implants.METHODS: HA orbital implants (250 cases) in our hospital (68 pegged implants) were reviewed.All patients were followed up from 18 months to 10 years. Implants were removed after medical therapy which was proved to be ineffective.RESULTS: Ten of 250 cases of HA orbital implants developed pyogenic granuloma. Pyogenic granuloma occurred in 1 unpegged implants patient and 9 patients after pegging and drilling of HA implantation over 4～7 years. The pyogenic granulomas were not controlled by medical therapy effectively. Implants were removed in 9 cases except 1 case denied removing and continued medical therapy.CONCLUSION: Pyogenic granuloma was serious complication that occurred after HA orbital implants. Partial vascularization, implant exposure, xenogenic sclera implant, pegging and drilling of HA implantation are risk factors that affect the development of pyogenic granuloma.Pyogenic granuloma hasn't relation with implanted peg material. Pyogenic granuloma denotes the potential implant infection, and all implants should be removed finally.

Porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus type 2 (PCV2) are very two important pathogens that have coursed huge economic losses in swine production in worldwide. In this study,a vector pCMV-TJM containing the full-length cDNA clone of PRRSV attenuated strain TJM-F92 was firstly constructed by PCR method. Then a gene sequence containing Afl II/Mlu I e restriction enzyme sites and a transcription regulatory sequence for ORF6 (TRS6) was inserted be- tween ORF7 and 3'UTR, yielding a expression vector pCMV-TJM-TRS. Subsequently, a plasmid pCMV-TJM-Cap was constructed by cloning of PCV2 ORF2 gene into the unique sites Afl II /Mlu I of pCMV- TJM-TRS plasmid DNA. Then three recombinant PRRSV, rTJM, rTJM/TRS and rTJM/Cap, were rescued by transfection of pCMV-TJM, pCMV-TJM-TRS and pCMV-TJM-Cap into Marc-145 cells, respectively,and confirmed by the genome sequence, restriction enzyme digestion, Western Blot and IFA. They all had the molecular markers which was different from the parent virus. The growth characteristics of the rescued viruses were similar to that of parent virus. rTJM/Cap could also express efficiently PCV2 Cap protein in Marc-145 cells. At passage 8, it still had PCV2 ORF2 gene which examined by RT-PCR. It indicated that the full-length cDNA clone of PRRSV attenuated strain TJM-F92 and recombinant PRRSV rTJM/Cap expressing PCV2 Cap protein were successfully constructed. It made an important foundation for studying on the pathogenic mechanisms of PRRSV and PRRSV-PCV2 vaccine in the future.
Animals ; Capsid Proteins ; genetics ; immunology ; Cell Line ; Circoviridae Infections ; veterinary ; virology ; Circovirus ; classification ; genetics ; metabolism ; Gene Expression ; Open Reading Frames ; Porcine Reproductive and Respiratory Syndrome ; virology ; Porcine respiratory and reproductive syndrome virus ; genetics ; metabolism ; Recombination, Genetic ; Swine ; Swine Diseases ; virology ; Viral Vaccines ; genetics ; immunology

ObjectiveToinvestigatetheplasmidgenechangesinquinolone‐sensitivePseudomonasaeruginosa.Methods 31iso‐lates from January 2011 to December 2013 from various qualified clinical samples in the hospital were collected .In the 31 isolateds , 16 isolates proved sensitive to quinolones by using K‐B method were used as research objects in the study .The isolates growing ou‐side the sensitive ring of ciprofloxacin paper were selected to continuously transferred into other culture dishes until the resistance to quinolones were acquired .Plasmid transformation and extraction were performed on those isolates to confirm the existence of drug‐resistanceplasmidsacquired,andthroughPCRandgenesequenceanalysistodeterminethetypeofplasmids.Results 2iso‐lates of quinolone‐sensitive Pseudomonas aeruginosa acquired drug‐resistance plasmids qnrS and were resistant quinolones induced by continuous transferring for 9 times .Conclusion If antibiotics of inhibitory concentration were often used for the treatment of Pseudomonas aeruginosa infection ,drug‐resistance plasmids were acquired easily .

OBJECTIVE: To analyze the chief factors influencing the adverse drug reactions (ADR) in outpatient use of antibiotics so as to formulate corresponding policies for intervention. METHODS: The patients who had been treated i.v with antibiotics from Mar. 2006 to Mar. 2007 were enrolled: 105 in trial group showed ADR, another 105 in control group showed no ADR. The ADR influencing factors were compared between the two groups from aspects of patients, nursing, and medication to find out the significant differences. RESULTS: The chief factors influencing the adverse drug reactions (ADR) in outpatient use of antibiotics included the indications of drugs, dosage, dosing interval, drug combination, and availability of medication guidance. CONCLUSION: The outpatient intravenous use of antibiotics is far from rational, which needes further intervention.

OBJECTIVE To investigate the distribution and bacterial resistance of nosocomial infection.METHODS The data of 45 hospitals from Shandong Provincial Nosocomial Surveillance System from Jan 2003 to Dec 2005 were analyzed.RESULTS Of total 5 626 isolates strains from the nosocomial infection cases,G-bacilli,G+ cocci and fungi accounted for 58.27%,25.84% and 15.89%,respectively.The ampicillin-resistant rate of commonly encountered G-bacilli was above 89%.There were 72.98% of E.coli resistant to ciprofloxacin.The rates of resistance of S.aureus and coagulase negative Staphylococcus to penicillin,ampicillin and erythromycin were all above 80%;the lincomycin-resistant rate of S.aureus increased gradually to 86.64%.CONCLUSIONS Drug resistance of the nosocomial infective bacteria is a serous problem.Surveillance of bacterial resistance should be strengthened.

Objective To determine whether macrophages can behave as antigen presenting cells participating the formation of immunological synapse in rheumatoid arthritis (RA) and whether this process can affect the apoptosis. Moreover, this study was aimed to observe the function of cyclophilin A (CypA) in immunological synapse formation and its role in regulating the apoptosis of macrophages. Methods human acute monocytic leukemia cell line (THP-1) induced macrophages were coated with staphylococcal enterotoxin B(SEB) (100 ng/ml) and co-cultured with activated Jurkat T cells (human acute T-cell leukemia cell line), then incubated in the RPMI-1640 for 16 hours to induce apoptosis. The apoptosis of the macrophages were analyzed by flow cytometry by Annexin V-PI staining. The macrophages cultured in the RPMI-1640 alone were used as control. Meanwhile, CypA (200 ng/ml) were added to or not added in order to observe the apoptosis of macrophages. The function of CypA and the apoptosis of macrophages isolated from RA peripheral blood were also investigated through co-culture with CD4+T cells isolated by immunomagnetic beads. Comparisons between groups were performed by two-sample t-tests. Results In the peripheral blood of healthy people and RA patients, the apoptosis of macrophages which participated immunological synapse was (32.9±2.8)%, (24.7±1.6)%, (14.5±1.2)% respectively, which was significantly lower than the apoptosis of macrophages cultured alone [ respectively for (61.4±2.4)%, (45.5±2.6)%, (22.9±1.5)%, (P<0.05) ]. After CypA was added, the apoptosis of macrophages in cell lines, healthy people and RA patients decreased to (27.2±2.1)%, (20.1±1.1)%, (12.9±1.0)%, lower than the apoptosis of macrophages which participated immunological synapse formation (P<0.05). Conclusion In RA, the macrophages participate in the formation of immunological synapse by interacting with CD4+ T cells. They can significantly reduce the apoptosis on themselves. CypA can enhance this effect. These results provide a new theoretical foundation for prolonged survival of macrophages in RA, which can secrete a variety of cytokines to enhance inflammation and joint destruction.