ObjectiveTo generate a prokaryotic expression system of series predominant epitopes (UreB322 and UreB527) of Helicobacter pylori UreB protein,and to synthesize a multiple antigenic peptide (MAP) vaccine by linking both the two epitopes with a peptide carrier (Poly-Asp-Lys),and to determine the immunogenicity and immunoprotection of the MAP vaccine.MethodsLinking primer PCR was performed to generate an enterokinase(EK) site-containing series UreB322 and UreB527 epitope encoding gene for construction of its prokaryotic expression system.The expressed target recombinant fusion protein 8 ×[rEK-UreB322-EK-UreB527-EK] was hydrolyzed with EK and then rUreB322-EK and rUreB527-EK epitope peptides were extracted using a Sephadex G-25 column.rUreB322-EK,rUreB527-EK and Poly-Asp-Lys were linked using carbodiimide method to produce a MAP vaccine (MAP-rUreB322/B527).The antigenicity and immunoreactivity of each of the two epitope peptides and MAP-rUreB322/527 were determined by ELISA and Western blot assay.An animal H.pylori strain SS1-infected model in BALB/c mice was used to detect the immunoprotection of MAP-rUreB322/527.ResultsAn octuple-repeated series UreB322-UreB527 encoding gene and its prokaryotic expression system were obtained.The yield of target fusion protein 8×[rEKUreB322-EK-rUreB527-EK] was as high as 48％ of the total bacterial proteins.EK hydrolyzed the target fusion protein completely into rUreB322-EK and rUreB527-EK peptides.The linking ratio of rUreB322-EK,rUreB527-EK and Poly-Asp-Lys was as high as 92.5％.The antibody against whole cell of H.pylori and rUreB-IgG could recognize and combine with the rUreB322-EK,rUreB527-EK or MAP-rUreB322/527.The specific serum antibody level in MAP-rUreB322/527-immunized mice was significantly higher than that in rUreB-immunized mice (P＜0.05).The immunoprotective rates(83.3％ and 91.7％ ) by immunization with50 or 100 μg MAP-rUreB322/527 in the H.pyloristrain SSI-infected mice were significantly higher that those(d1.7％ and 50.0％ ) by immunization with equal rUreB(P＜0.05).ConclusionAn gene composed for encoding a repeated series predominant epitopes of H.pylori UreB protein and its prokaryotic expression system are successfully generated in this study.MAP-rUreB322/527,the multiple antigenic peptide vaccine based on the two predominant epitopes of UreB,can noticeably increase the immunoprotection in H.py/or/infected mice.

Animals ; Cattle ; China ; epidemiology ; Echinococcosis ; epidemiology ; parasitology ; transmission ; Echinococcus granulosus ; pathogenicity ; Humans ; Seasons

Animals ; Conditioning, Classical ; Evoked Potentials, Somatosensory ; Fear ; psychology ; Male ; Rats ; Rats, Sprague-Dawley ; Stress, Psychological

Background Many eye diseases such as central retinal artery occlusion,glaucoma and ischemic optic neuropathy,etc.lead to retinal ischemia-reperfusion injury (RIRI) and furthmore visual functional damage.It is neeessary to study the treatment of RIRI.Objective This study was to observe and discuss the influence of aminoguanidine on the retina morphological changes and its mechanism after RIRI.Methods Eighty clean healthy male Japanese white rabbits were randomly divided into normal injury group,RIRI group and aminoguanidine (AG)treated group.The model of RIRI was established by infusing saline solution into the anterior chamber to elevate intraocular pressure (IOP) in both RIRI group and AG group.AG was intraperitoneally injected in the models of the AG group,and normal saline solution was used at the same method in tbe normal group and the RIRI group.The fundus photography and fundus fluorescein angiography(FFA) were pertormed on the rabbits at the moment of retina ischemia and 6,24 and 72 hours after reperfusion.The parts of rabbits were sacrificed 1,6,24 and 72 hours after reperfusion,followed by the enucleation of the eyeballs.Retinal section was prepared for TUNEL examination to evaluate the apoptosis of retinal cells.Nitric oxide (NO) concentration in retina was detected with nitrate reductase,and the activity of inducible nitric oxide synthase (iNOS) was measured by colorimetric detection.The use of the animals followed the Regulations for the Administration of Affairs Concerning Experimental Animals by State Science and Technology Commission.Results The fundus photography and FFA showed that the retinal edema was more mild,and the percentage of vascular occlusion was lower in the AG treatment group than that in RIRI group and the amount and area of fluorescein leakage were also smaller than the treatment group.The numbers of TUNEL positive cells in the AG treatment group were less than those in the RIRI model group at 1,6,24 and 72 hours after experiment (F分组 =2762.37,P =0.00 ; F时间 =894.24,P =0.00).Numbers of TUNEL positive cells between adjacent time points were significantly different in both RIRI model group and AG treatment group (RIRI group:q =24.475,36.591,-20.37,P＜0.05;AG group:q =20.94,16.79,-6.92,P＜0.05),with the peak value at 24 hours after experiment.NO contents were significantly higher in the RIRI model group compared to AG group at various time points(q =3.84,4.01,8.91,3.75,P＜0.05),and those between adjacent time points showed significant differences (RIRI group:q=4.77,13.403,-10.29,P＜0.05;AG group:q=4.55,9.05,-5.08,P＜0.05).iNOS activity was significantly elevated in the RIRI model group compared with AG group(q=-3.74,-4.94,-6.53,-3.98,P＜0.05),and obvious differences also were seen between the adjacent time points in both two groups(RIRI group:q =8.43,6.71,-6.39,P＜0.05 ;AG group:q =4.16,5.08,-3.93,P＜0.05).Conclusions Aminoguanidine can protect the retinal function and morpbology from oxidative stress damage after RIRI by reducing the NO level and inhibiting the iNOS activity in retina.

Calcinosis ; pathology ; Child ; Humans ; Inhibins ; metabolism ; Male ; S100 Proteins ; metabolism ; Sertoli Cell Tumor ; metabolism ; pathology ; surgery ; Testicular Neoplasms ; metabolism ; pathology ; surgery ; Vimentin ; metabolism

Objective To explore the anti-inflammatory and antipruritic effects of Chushizhiyang ointment in a mouse model. Methods A total of 40 male 8-week-old BALB/c mice were included in this study, and randomly and equally divided into 4 groups. A mouse model of atopic dermatitis (AD)was established in three groups of mice by repeated application of 2,4-dinitroflurobenzene (DNFB)to shaved abdominal skin for sensitization and to shaved dorsal skin for stimulation. After establishment of the AD model, the three groups were topically treated with sodium chloride physiological solution (model group), hydrocortisone cream (hydrocortisone group)and Chushizhiyang ointment (Chushizhiyang group)respectively for 14 consecutive days. The remaining group receiving no sensitization or treatment served as the normal control group. All the mice were sacrificed 12 hours after the final treatment, and the dorsal skin of mice was resected followed by the determination of skin thickness and weight as well as hematoxylin-eosin(HE)staining and toluidine blue staining for the counting of leukocytes and mast cells respectively. Moreover, enzyme-linked immunosorbent assay(ELISA)was performed to measure the levels of interferon-gamma(IFN-γ), tumor necrosis factor alpha(TNF-α), interleukin 4(IL-4)and IL-5 in dorsal skin lesions. In addition, a local skin itching model was induced by histamine phosphate in Hartley guinea pigs, which was used to explore the effect of Chushizhiyang ointment on itch thresholds. Results Compared with the model group, both the Chushizhiyang group and hydrocortisone group showed reduced thickness and weight of dorsal skin in mice (all P < 0.01), numbers of infiltrating lymphocytes and mast cells (all P < 0.01)and levels of IFN-γ, TNF-α, IL-4 and IL-5 in skin lesions (P < 0.05 or 0.01)on day 15 after the start of treatment. The thickness and weight of dorsal skin in mice were significantly decreased in the hydrocortisone group (P <0.01), but experienced no significant changes in the Chushizhiyang group compared with the normal control group. Additionally, Chushizhiyang ointment could significantly increase itch thresholds in guinea pigs induced by histamine phosphate(P < 0.01). Conclusions Chushizhiyang ointment can significantly inhibit DNFB-induced AD in mice, likely by restoring the balance between Th1 and Th2 type cytokines. Moreover, Chushizhiyang ointment could markedly relieve itching induced by histamine phosphate in guinea pigs.

Aim To establish and optimize the VSVG/HIV-1NL4-3 Luc pseudovirus model for anti-HIV drugs screening. Methods The infectivity of VSVG/HIV-1 NL4-3 Luc in 4 different cell lines was investigated according to the method of the lucifer-ase activity analysis system of Promega company. 3 different ex-perimental settings were used to detect the activities of approved anti-HIV drugs to confirm the feasibility and effectiveness of the system. Finally, some potential compounds were screened for their anti-HIV activities, and their antiviral activities against the pseudovirus were compared with HIV-1ⅢB . Results The pseud-ovirus showed the strongest replication ability in CRFK cells, and a clear dose-effect relationship was found between the report gene expression level and the virus quantity. Comparing the EC50 of different positive inhibitors against VSVG/HIV-1 NL4-3 Luc on 3 kinds of experimental conditions, 3rd scheme is the best. Finally, the system was used to screen compounds, the EC50 s a-gainst pseudovirus were similar to those in HIV-1ⅢB . Conclusion An optimized VSVG/HIV-1 NL4-3 Luc anti-HIV screening sys-tem has been successfully developed.

ObjectiveTo observe the prepulse inhibition(PPI) ot the startle reflex of pure cerebral concussion (PCC) suffered from one concussion and multiple cerebral concussion (MCC) suffered from three concussions in rats,and to explore accumulate effect upon cognitive dysfunction of MCC.MethodsA metallic pendulum striker device for closed head injury was employed to duplicate PCC and MCC models in Stragu-Dawley rats.The MCC rats were hit three times on rats'head and it is interval 24 hours for every hit.According to the criteria of cerebral concussion,the investigated animals were divided into PCC group and MCC group at freedom.One control group was used.Each group included 10 animals.Each experience mental animal was tested from 3 days pre-injury to 28 days post-concussion.Startle reflex amplitude (for P values),pre-stimulation induced reflex amplitude on three standard stimulations,that was,67dB,69dB and 73dB (for PP67,PP69 and PP73 values) and prepulse inhibition of the startle reflex (PPI) were collected.ResultsThe P values and three PP values in the first three days of pre-replication experiment,there was no statistical significance in each group.However the P values and PP67,PP69 and PP73 values declined until to the 16th day after injury (P＜0.05),then recovered in PCC group.The P value and PP67,PP69 and PP73 values changes of MCC group declined and not recovered until to test end (P＜0.05 ) and they were more lower than PCC.The three PPI values were a little bit increase in both groups,there were statistics significance at some test points (P＜0.05) compared with control.ConclusionThe startle reflex amplitude and pre-stimulation induced reflex amplitude weaken after cerebral concussion and there is damaging accumulate effect to injury times,the PPI is enlanced by cerebral concussion.

Aniline Compounds ; blood ; Gas Chromatography-Mass Spectrometry ; Humans ; Sensitivity and Specificity

Objective To discuss the clinical value of 99Tcm-sestamibi(99Tcm-MIBI) myocardial perfusion imaging on detecting myocardial ischemia in children with Kawasaki disease(KD) at convalescence period.Methods Twenty-one children wih KD at convalescence period were divided into 2 groups according to results of echocardiography.Four cases with coronary artery dilation,17 cases without coronary artery dilation.All cases accepted dipyridamole stress myocardial perfusion imaging.These patients who had positive results were given rest myocardial perfusion imaging again next day.Results Among 21 cases,9 cases(42.8%) were positive in perfusion imaging.Four cases with coronary artery dilation showed myocardial ischemia in different degree detected by myocardial perfusion imaging.Among 17 cases without coronary artery dilation,5 cases(29.4%) were positive.Conclusions Compared to echocardiography,99Tcm-MIBI myocardial perfusion imaging can objectively evaluate the location,extent and degree of myocardial ischemia of children with KD.It will be a routine test in observing its phase development.