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.

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

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

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

Objective To application of solution focused approach in the functional gastrointestinal disorders.Methods 64 hospitalized clients with functional gastrointestinal disorders were randomly divided into the observation group and the control group with 32 cases in each group.We applied the normal nursing intervention to the control group,while applied the solution focused approach to the observation group.The anxiety and depression score,score of coping way and gastrointestinal symptoms were evaluated using t test.Results The scores of anxiety and depression,coping way and gastrointestinal symptoms were better in the observation group than those in the control group.Conclusions Adoption of solution focused approach can relieve FGID sufferers'negative emotion,such as anxiety and depression,increase sufferers' coping capacity and symptoms,which demonstrate pivotal significance for rehabilitation of functional gastrointestinal disorders.

Calcium, as a crucial signal molecule, plays an important role in the development process of immune cells and their functions. After T cell receptors or B cell receptors are stimulated or co-stimulated, the signaling pathways lead to Ca2+ influx and induce the increase of Ca2+ level in cells. Ca2+ signals have different short-time and long- term functions on T and B lymphocytes. Many immune deficiency disorders are resulted from the decrease of Ca2+ influx. This article reviewed the effects of calcium on the functions of T and B lymphocytes and its relationship with some immune deficiency disorders.

BACKGROUND: Previous studies have demonstrated that the responses of myocardium to short periods of ischemia/reperfusion are associated with changes in the expressions of a variety of genes. However, the characteristics of these changes are unknown.OBJECTIVE: To clone and analyze a stress-inducible gene characteristics in order to study the molecular mechanism of myocardium to short periods of ischemia/reperfusion damage.DESIGN: A controlled animal study.SETTING: Department of Hematology, Xiangya Hospital of Central South University.MATERIALS: Sixteen castrated male German Landrace-type domestic porcines, weighing between 21 and 39 kg, were provided by the laboratory of Department of Experimental of Cardiology at Bad Nauheim in Germany. The porcines were randomly divided into two groups ischemia/reperfusion group (n =14) and control group (n =2). The porcines in the ischemia/reperfusion group were observed at 0, 30 and 90 minutes of reperfusion after reocclusion, and ischemia and non-ischemia myocardial tissues were harvested from each pocrine.METHODS: The experiments were performed in the laboratory of Department of Experimental Cardiology of Max-Planck Institute for Physiological and Clinical Research at Bad Nauheim in Germany and the Department of Hematology, Xiangya Hospital of Central South University from July in 1998 to May in 2007. All the animals were anesthetized and thoracotomized. Following 30 minutes of stabilization, the left anterior descending coronary artery (LAD) was occluded for 10 minutes followed by 30-minute reperfusion and then another 10-minute reocclusion. The porcines were killed immediately at corresponding time points. The sham-operated animals were killed without occlusion. Experimental tissue was removed from the LAD area and control tissue from the region of the left circumflex coronary artery area. Firstly, a porcine heart cDNA library was screened, DNA and deduced amino acid sequences were then analyzed. Meanwhile, short periods of myocardium ischemia/reperfusion was performed by occluding porcine LAD followed by reperfusion as mentioned above, and total RNAs isolated from myocardium or a variety of other tissues were used for Northern blotting. Quantitation of mRNA levels was accomplished by using a PhosphorImager and Image Quant software, the ration of mRNA/18S rRNA was suggested as the level of gene expression.MAIN OUTCOME MEASURES: ① DNA and amino acid sequences analysis of the cloned gene; ② Analysis of the cloned gene expression.RESULTS: All the 16 porcines were involved in the final analysis of results. After screening, a cDNA fragment with 3 461 base pairs was obtained. DNA sequencing and searching revealed that this cDNA shared 86% identity to human structural maintenance of chromosome 6 gene (hSMC6), and 84% identity to Mus musculus SMC6 (mSMC6). Furthermore, a largest polypeptide deduced from this cDNA contained 1007 amino acid residues, and protein homology searching indicated that the predicted polypeptide carried 92% and 89% identity to hSMC6 or mSMC6 protein respectively. Thus, the cloned cDNA was referred to as a porcine homology gene to hSMC6 (pSMC6). The results of Northern blotting showed that the expression of the pSMC6 mRNA in ischemia/reperfusion myocardium was increased, and its mRNA existed in all organs tested. CONCLUSION: A stress-inducible gene of pSMC6 from porcine myocardium has been cloned, which is high homology to hSMC6 gene, and the pSMC6 or along with other molecules is possible involved in the early repair of myocardium to DNA damage caused by ischemia/reperfusion in the porcine heart.

Objective To observe the effect of nasal continuous positive airway pressure (nCPAP) and Biphase positive airway pressure (BIPAP) on the oxygen exchange indexes in newborns with mild to moderate neonatal respiratory distress syndrome (NRDS).Methods After infusion of pulmonary suffactant into lung bilaterally,116 cases with mild to moderate NRDS were treated with nCPAP(nCPAP group,n =75) and BIPAP(BIPAP group,n =41) respectively.Oxygen exchange indexes including arterial partial pressure of oxygen [pa (O2)],modified oxygenation index (P/F),alveolar-arterial oxygen tension difference (A-aDO2),respiratory index (RI),artery/alveolar oxygen partial pressure ratio[pa (O2)/pA (O2)] and pulmonary shunt fraction (Qs/Qt) were measured or calculated at 0 (before treatment),2,8 and 24 h after being treated with nCPAP or BIPAP.Results Six different oxygen exchange indexes of Pa (O2),P/F,A-aDO2,RI,pa (O2)/pA(O2) and Qs/Qt in both groups didn't show statistical significance at 0 hour:pa(O2) (kPa) (6.1 ±0.6 vs 6.1 ±0.6,t =0.11,P ＞0.05),P/F(kPa) (15.59 ± 1.45 vs 15.71 ± 1.45,t =1.59,P ＞ 0.05),A-aDO2(kPa)(6.04 ±0.64 vs 6.24 ±0.69,t =1.59,P ＞0.05),RI(6.0 ± 1.0 vs 6.2 ± 1.1,t =1.35,P ＞ 0.05),p,(O2)/pA(O2) (0.24 ±0.02 vs 0.25 ±0.03,t =1.63,P ＞0.05)and Qs/Qt(％)(11.9±1.6 vs 11.6 ± 1.6,t =1.10,P ＞ 0.05).A-aDO2,RI,pa (O2)/pA (O2) and Qs/Qt of BIPAP group had more improvement than those of nCPAP at 2 and 24 h,and oxygen exchange indexes at 2 h of BIPAP group and those of nCPAP group were:A-aDO2 (kPa) (3.83 ±0.49 vs 4.24 ± 0.67,t =18.26,P ＜ 0.05),RI(2.7 ± 0.5 vs 3.3 ±0.7,t =20.59,P ＜ 0.05),p,(O2)/pA(O2) (0.35 ±0.03 vs 0.32 ±0.04,t =15.35,P ＜0.05) and Qs/Qt(％) (8.8 ± 1.6 vs 9.8 ±2.0,t =7.68,P ＜ 0.05) ; 24 h indexes in BIPAP group and the indexes of nCPAP group were:A-aDO2 (kPa) (2.29 ± 1.19 vs 3.07 ± 1.67,t=18.43,P ＜0.05),RI(1.4 ±1.4 vs 2.3 ± 1.6,t=25.02,P ＜0.05),pa(O2)/pA(O2) (0.49 ±0.10 vs 0.42 ±0.11,t =14.96,P ＜0.05) and Qs/Qt(％) (6.5 ±3.0 vs 8.5 ±4.4,t =9.59,P ＜0.05).pa(O2) and P/F of both groups didn't show statistical difference significantly [Pa (O2)(kPa) (12.6 ± 1.0 vs 12.7 ± 1.0,t =3.76,P ＞ 0.05),P/F(28.49 ± 3.17 vs 31.85 ± 2.85,t =3.76.P ＞ 0.05)].Nineteen cases in nCPAP group needed invasive mechanical ventilation,and only 3 cases in BIPAP group underwent the procedure;there was a significant difference (x2 =4.01,P ＜ 0.05).Oxygen exchange indexes in both groups were gradually improved from 2 h,8 h to 24 h,but 22 deteriorated cases showed no improvement,who underwent invasive ventilation.Positive end-expiratory pressure (PEEP) positive airway pressure (EPAP) was (0.42 ± 0.19) kPa in BIPAP group,and (0.56 ± 0.23) kPa in nCPAP group (t =3.45,P ＜ 0.01).Conclusions 1.BIPAP and nCPAP can improve the ability of oxygen exchange in newborns with mild to moderate NRDS,but BIPAP is more effective than nCPAP,which can shorten the possibility of invasive ventilation and reduce the PEEP level.2.p,(O2),P/F,pa (O2)/pA (O2) and RI may be more suitable for evaluating oxygen exchange for neonatal NRDS.