Bacteriophages are viruses that infect microorganisms such as bacteria,fungi,actinomycetes and spirochetes.Because of the inherent immunogenicity,genetic plasticity,stability,safety and many other advantages,it has unique potential in vaccine research and development. At present,there are countless researches using it to construct vaccine delivery platforms,mainly including three forms,phage display vaccine,phage DNA vaccine and hybrid phage DNA vaccine,of which the phage display vaccine is the most widely studied. Phage display technology is a novel vaccine preparation technology,which is a molecular biology technology using phage as carrier,integrating foreign polypeptide or protein genes into phage genes and displaying them on the surface of phage in the form of fusion protein. This review mainly elaborated the immunological basis of phage display vaccine,the display system and its application in disease prevention,so as to provide a reference for the development and application of phage display vaccine.

Objective To construct a recombinant M13 phage vaccine targeting the outer membrane protein P6 of nontypeable Haemophilus influenzae(NTHi) and evaluate its immunogenicity in order to provide new ideas for further development of NTHi vaccines. Methods The NTHi P6 gene was fused with the vector pMECS Phagemid by gene recombination technique.After packaging and purification, the obtained recombinant P6-M13 phage was prepared into recombinant P6-M13 phage vaccine. The expression of P6-M13 PⅢ fusion protein in the vaccine was detected by Western blot, the vaccine titer was determined by double-layer agar plate method, and the recombinant P6-M13 phage morphology was observed under transmission electron microscope. Ninety 5-week-old BALB/c female mice were randomly divided into PBS group, M13 phage group and recombinant P6-M13 phage vaccine group, 30 for each, and intraperitoneally injected with PBS(500 μL/mouse), M13 phage[1 × 10~(12)pfu/(500 μL·mouse)]and recombinant P6-M13 phage vaccine[1 × 10~(12)pfu/(500 μL·mouse)]on the 0, 14th and28th day, separately. Two weeks after the last immunization, the levels of specific IgG in serum and IFNγ, IL-2, IL-4, IL-5and IL-17A in spleen lymphocyte culture supernatant were detected by ELISA, and the proliferation of spleen lymphocytes was analyzed by CCK-8. Three weeks after the last immunization, the mice were challenged with 1. 5 × 108cfu/mL NTHi through the nasal cavity. After one week of challenge, the pathological changes of nasal mucosa and lung tissue were observed by HE staining. Four weeks after the last immunization, the heart, liver, spleen, lung and kidney of mice were weighed, the organ coefficients were calculated, and histopathological sections were prepared for pathological observation.Results The recombinant P6-M13 phage could correctly express P6-M13 PⅢ fusion protein with a titer of 5. 5 × 10~(14) pfu/mL,and the recombinant P6-M13 phage with regular morphology was observed under microscope. Compared with M13 phage group and PBS group, the level of serum specific antibody IgG in mice of recombinant P6-M13 phage vaccine group was significantly higher(F = 71. 489, P < 0. 05); the levels of IFNγ, IL-2, IL-4 and IL-5 secreted by mouse spleen cells decreased significantly(F = 8. 315, 16. 986, 39. 204 and 6. 291, respectively, each P < 0. 05), while there was no significant difference in IL-17 level among the three groups(F = 0. 863, P > 0. 05); the spleen cell stimulation index increased significantly(F =22. 952, P < 0. 05). After challenge, the nasal mucosa and lung tissue structures of mice in PBS group and M13 phage group were seriously damaged, and inflammatory cells increased, while in the recombinant P6-M13 phage vaccine group, the structures of nasal mucosa and lung tissue were normal with few inflammatory cells. There was no significant difference in the organ coefficients of heart, liver, spleen, lung and kidney of mice in each group(F = 1. 012, 1. 642, 0. 300, 2. 079, and 0. 405, respectively,each P > 0. 05), and no pathological changes were found in the general color morphology and pathological sections of the main organs. Conclusion The constructed recombinant P6-M13 phage vaccine targeting NTHi outer membrane protein P6 can induce effective humoral and cellular immunity in mice with certain immune protection ability and good safety.

De Lange Syndrome ; Humans ; Infant, Newborn ; Infant, Premature

Fractal, a mathematics concept, is used to describe an image of self-similarity and scale invariance. Some organisms have been discovered with the fractal characteristics, such as cerebral cortex surface, retinal vessel structure, cardiovascular network, and trabecular bone, etc. It has been preliminarily confirmed that the three-dimensional structure of cells cultured in vitro could be significantly enhanced by bionic fractal surface. Moreover, fractal theory in clinical research will help early diagnosis and treatment of diseases, reducing the patient's pain and suffering. The development process of diseases in the human body can be expressed by the fractal theories parameter. It is of considerable significance to retrospectively review the preparation and application of fractal surface and its diagnostic value in medicine. This paper gives an application of fractal and its theories in the medical science, based on the research achievements in our laboratory.
Biomedical Research ; Bionics ; Fractals ; Humans

Changes in heart rate of fetal is function regulating performance of the circulatory system and the central nervous system, it is significant to detect heart rate of fetus in perinatal fetal. This paper puts forward the fetal heart rate detection method based on short time Fourier transform and blind source separation. First of all, the mixed ECG signal was preprocessed, and then the wavelet transform technique was used to separate the fetal ECG signal with noise from mixed ECG signal, after that, the short-time Fourier transform and the blind separation were carried on it, and then calculated the correlation coefficient of it, Finally, An independent component that it has strongest correlation with the original signal was selected to make FECG peak detection and calculated the fetal instantaneous heart rate. The experimental results show that the method can improve the detection rate of the FECG peak (R), and it has high accuracy in fixing peak(R) location in the case of low signal-noise ratio.
Electrocardiography ; Fetus ; Heart Rate, Fetal ; Humans

Objective To identify a potential correlation between cold hemoagglutinin(CHA) and diffuse panbronchiolitis(DPB) in Chinese patients.Methods Eighteen patients diagnosed as DPB from December 1996 to July 2008 in Peking Union Medical College Hospital and 60 cases of DPB reported in mainland of China from 1996 to 2008 were enrolled in the study.Results Of 18 patients diagnosed as DPB in Peking Union Medical College Hospital,only one patient showed a titer of CHA≥1:64.Of 60 cases in mainland China,48 cases were CHA positive.CHA was positive in 54.1% all cases.There may be some correlation between positive rate of CHA and medication as well as population.Conclusion Low positive rate of CHA in Chinese subjects,which is different from that of Japanese DPB patients,suggests that CHA may not be applied as a diagnostic criteria for Chinese patients.

Choroidal neovascularization(CNV)is one of common causes of vision loss.The pathogenesis and development of CNV are a comprehensive process which is regulated by multiple factors and cytokines.Cyclooxygenase-2(COX-2)is an inducible isoform of cyclooxygenase and rate limiting enzyme in the prostaglandin biosynthesis pathway.COX-2 plays an important role in neovascularization by modulating vascular endothelial growth factor(VEGF),migration and apoptosis of endothelial cell.Recently,some experimental studies demonstrated that COX-2 involves in the information of CNV and the inhibitor of COX-2 can suppress CNV.These results provide a new prospect for the prevention and treatment of the CNV.Biological characteristics of COX-2 and its relationship with CNV are reviewed in this article.

Objective To investigate the susceptibility and resistance profile of clinical isolates.Methods Clinical isolates were collected from Children's Hospital of Chongqing Medical University from January 1 to December 31,2015.Antimicrobial susceptibility testing was carried out according to a unified protocol using Kirby-Bauer method or automated systems.Results were analyzed according to CLSI 2015 breakpoints.Results A total of 13 109 clinical isolates were collected from January to December 2015,of which gram negative organisms and gram positive cocci accounted for 65.3 ％ (8 560/13 109) and 34.7 ％ (4 549/13 109),respectively.Methicillin resistant strains in S.aureus (MRSA) and coagulase negative Staphylococcus (MRCNS) accounted for 29.6 ％ and 67.1％,respectively.Most (93.1％) MRSA strains were still susceptible to trimethoprim-sulfamethoxazole,while 80.2 ％ of MRCNS strains were susceptible to rifampin.No staphylococcal strains were found resistant to vancomycin,teicoplanin or linezolid.The resistance rates of E.faecalis strains to most antibiotics tested (except tetracycline) were much lower than those of E.faecium.Some strains of both species were resistant to vancomycin.No E.faecalis or E.faecium strains were found resistant to vancomycin.The prevalence of ESBLs-producing strains was 55.7 ％ in E.coli and 43.5 ％ in Klebsiella (K.pneumoniae and K.oxytoca) and 11.6 ％ in Proteus mirabilis isolates.ESBLs-producing Enterobacteriaceae strains were more resistant than non-ESBLs-producing strains in terms of antibiotic resistance rates.Enterobacteriaceae strains were still highly susceptible to carbapenems.Overall,less than 16.0 ％ of these strains were resistant to carbapenems.About 10.5 ％ and 9.4 ％ of the A.baumannii strains were resistant to imipenem and meropenem,respectively.Compared to the data of year 2014,the prevalence of extensively-drug resistant P.aeruginosa and K.pneumoniae strains increased.Conclusions The antibiotic resistance of clinical bacterial isolates is growing.The emerging and increasing prevalence of multi-drug or pan-drug resistant strains poses a serious threat to clinical practice and implies the importance of strengthening infection control.

Oxidative stress plays an important role in the progression of diabetes mellitus. As a potent antioxidant, alpha-lipoic acid is able to clear free radicals and alleviate oxidative damages and therefore has been widely applied in the clinical management of diabetes mellitus. This article summarizes the clinical application of alpha-lipoic acid in alleviating diabetes mellitus-related oxidative damages, protecting vascular lesions, treating diabetic polyneuropathies, and modulating insulin sensitivity.