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.

Objective To compare the antiviral efficacy and economics between lamivudine and entecavir in patients with low preoperative HBV replication after liver transplantation. Methods Liver transplantation patients from 2006 July to 2013 July were selected for being studied prospectively. Statistically evaluated between entecavir and lamivudine efficacy by the analysis of postoperative follow-up of seroconversion and HBV reinfection. Results The cumulative overcast rate of entecavir group was 98.04% one month after transplatation. And 3 of 51 cases were reinfected with average turn time of 9.73 days. Meanwhile , the cumulative overcast rate of Lamivudine group was 100% and 3 of 26 cases were reinfected with average turn time of 5.11 days. There are no statistical differences between the two groups accumulation rate , reinfection rates , as well as cumulative infection rates. Conclusion In the cases of low hepatitis B virus replication before liver transplantation, Entecavir and Lamivudine have the similar effect of preventing HBV reinfection after operation. Compared with the cost of Entecavir group, Lamivudine is more economy.

Objective To observe the clinical efficacy of intradermal needle therapy in treating chronic pharyngitis.Method Sixty patients with chronic pharyngitis were randomized into a treatment group and a control group, 30 cases in each group. The treatment group was intervened by intradermal needle therapy at Lieque (LU 7) and Zhaohai (KI 6), while the control group was by Compound Borax solution plus Cydiodine Buccal tablets. The symptoms were scored before and after treatment, and the clinical efficacies were compared.Result The total effective rate was 93.3% in the treatment group versus 80.0% in the control group, and the difference was statistically significant (P<0.05). The symptom scores were markedly changed after the treatment in both groups (P<0.01). The symptom score of the treatment group was significantly different from that of the control group (P<0.05). Conclusion Intradermal needle therapy is an effective method in treating chronic pharyngitis.

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.

Objective To observe the treatment effect of transplanted autogenous bone marrow stroma cells (BMS-C) absorbed in the type-Ⅰ collagen matrix in bone repair. Methods Thirty-two adult white New Zealand rabbits were randomly divided into Group Ⅰ and Group Ⅱ. The bone marrow was extracted from tubercle and greater trochanter of femur for culture. Then, the cells were amplified, implanted into type-Ⅰ collagen matrix and further cultured for two weeks. Two weeks later, the cells were implanted into the 1.5 cm radius defect. In the Group Ⅰ, management A (BMS-C+type-Ⅰ collagen) was given to the defect of one radius and management B (control) to the defect of the contralateral radius. In the Group Ⅱ, the defect of one radius was given management A and the defect of the contralateral radius management C (type-Ⅰ collagen).The rabbits were killed 8 and 12 weeks after implantation, the effect of three managements was compared. Results Twelve weeks after implantation, in the management A, all specimens had complete bony union; and in the management B, the specimens were filled with fibrous tissues; in the management C, a little scar formed. Conclusions BMS-C absorbed in the type-Ⅰ collagen can effectively repair bone defect.

The morphologc observation of immunoreactive structures and the quantitative analysis of VIP, SP, L-ENK and SOM in the small intestine of the control and the irradiated rats were studied by using the methods of immunocytochemistry (ICC) and radioimmunoassay (RIA). Meanwhile, histopathologic study was carried out by light and electron microscope (LM, EM). The results and findings are as follows.. (1) Viewed on LM, no obvious change of the enteric ganglia was found in the small intestine of the irradiated rats; while under EM, the axons and synaptic vesicles showing swelling and vacuolar degeneration was observed. These stuctures have close relation to the transport and release of the neurotransmitter. (2) The normal morphology and the distribution of the immunoreactive nerves and endocrine cells of VIP, SP, L-ENK and SOM were revealed in the intestine of rats by ICC method. Four kinds of immunoreactive peptidergic nerves were confirmed as an important part of the enteric nervous system. No obvious change was found on the structure of the peptidergrc nerves after 2000 rad r-irradiation. The radiosensitivity of the peptide-containing endocrine cells was similar to that of the intestinal epithelium. (3) We first confirmed with RIA that the level of the 4 regulatory peptides remarkably changed in the small intestine of the irradiated rats. Combining the physiologic founctions and the trends of the 4 regulatory peptides, as well as the pathologic characteristics and process after r-irradiation, it is proposed that the 4 kinds of peptide may be involved in the pathologic process of the mucosa damage and the disturbance of intestinal motility etc. in the intestinal radiation sickness of rats.

Since rabbit (Oryctolagus cuniculus) possess many fine biological characteristics, they have been used in the studies of tumour, gene mapping and evolution. All of these need us to know the high-resolution G-banding pattern of rabbit chromosomes. Primary lung fibroblasts of new born rabbits were cultivated in RPMI 1640 medium supplemented with 20% new born bovine serum. In order to get the synchronized cells, chemical(MTX, TdR) method and physical (Cold Shock) method were used. By using trypsin-Giemsa banding-technique, we obtained fine high-resolution G-banding karyotypes of rabbit. The bands of rabbit chromosomes were analyzed and described. An idiogram which contains 583 bands of high-resolution G-banding chromosomes of rabbit has been made. The feature of rabbit choromosome bands and the method uesd to get the high resolution banding chromosomes were briefly discussed.

Objective To establish a HPLC method for the determination of allantoin in Rhizoma Dioscoreae from different producing areas. Methods The RP- phase HPLC is applied. Separation and determination of allantoin was achieved by using an Allttima ODS C18 column(250 mm? 4.6 mm, 5? m) and a mobile phase of methanol- water (1 ∶ 39, V/V). The flow- rate was 0.8 mL/min, temperature at 30 ℃ and detection wavelength at 224 nm. Results A good linearity of allantoin was in the range of 0.24 ? g～ 2.40 ? g, r=0.9992; the average recovery rate was 103.1 % , RSD =2.0 % (n=6). Conclusion Rhizoma Dioscoreae from Henan Province is of the best quality and Rhizoma Dioscoreae from other provinces are only qualified for food. 