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.

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.

To evaluate the mechanism,treatment and prescription acupoint of acupuncture when it was used for osteoporosis.The observation index,effect and outcomes were studied objectively.The characteristics and associated problems of acupuncture were discussed.

Neonatal necrotizing enterocolitis is a common gastrointestinal disease in neonate.Recently,with the establishment of the NICU and the all-round development of perinatal medicine and pediatrics,survival rate of premature especially very low birth weight neonates have obviously increased and the incidence of the disease is rising year by year.It is generally considered that multiple risk factors such as premature birth,low birth weight,infections,asphyxia,irrational use of drugs and so on,acting on the immature intestine,but the pathogenesis is still unknown.Therefore,it is significant to clearify the pathogenesis of the risk factors of NEC for studying the pathogenesis of the disease and providing prevention and control strategy.

Objective To evaluate the efficacy and safety of Haijiyi under nasal endoscope in the treatment of allergic rhinitis(AR).Methods 213 cases with AR were randomly divided into treatment group(112 cases) and control group(101 cases).Patients in the treatment group were treated with Haijiyi under nasal endoscope.The control group was treated with traditional microwave therapeutic equipment.Both two groups were followed up for 1 year,and long-term clinical efficacy was observed and compared between the two groups.Results The total effective rate of the treatment group was 80.4％,which was higher than 61.4％ of the control group,and the difference was statistically significant(P＜ 0.01).Conclusion Haijiyi CZB type in the treatment of AR has advantages of exact allocation,no bleeding,less complication,good long-term efficacy,which is worth clinical popularizing and application.

Objective To investigate the influence of enteral nutrition (EN) via feeding pump-based continuous infusion (FPBCI) on nutrition support-associated complications in stroke patients. Methods Totally 53 patients were randomized into BPBCI group (n = 28) and control group (bolus feeding) (n = 25 ). The nutrition support-associated complications including bloating, diarrhea, hyperglycemia, and regurgitation-associated low respiratory tract infection were compared between these two groups. Results The incidences of nutrition support-associated complications were significantly lower in BPBCI group than in control group (bloating: 14.3%vs. 64.0%, P=0.0002; diarrhea: 25.0% vs. 52.0%, P=0.0429; hyperglycemia: 17.9% vs. 44.0%, P=0. 0385 ). There was no significant difference in regurgitation-associated lower respiratory tract infection between these two groups (32. 1% vs. 40.0% , P = 0. 5516). Conclusion Compared with conventional bolus feeding,BPBCI may greatly decrease tube-feeding related complications.

Perioperative nutrition support may be required when the synthesis and utilization of nutrients become disordered due to gastrointestinal abnormalities and trauma anesthesia. Proper nutrition support can reduce or prevent malnutrition, and thus facilitate the recovery and lower complications and case-fatality rate.

The bilateral idiopathic aldosteronism (IHA) is the most common subtype of aldosteronism.Spironolactone is the primary preferred agent due to the pathophysiology of IHA and the long-standing clinical experience over years.If patients faced with severe side effects of spironolactone or poorly controlled blood pressure by this first-line treatment,additional treatment alternative to spironolactone,second antihypertensive,as well as adrenalectomy are suggested.In recent years,several new agents were developed to treat IHA,such as aldosterone synthase inhibitor and eplerenone.This review introduces these new kinds of medicine in the treatment of IHA.