Objective To identify the immunogenicity and the potential of using Schistosoma japonicum mitochondria related protein(rSj338) as a candidate vaccine antigen for Schistosomiasis japonica . Methods The expressed recombinant fusion protein(rSj338/26GST) was purified and recognized by S.japonicum heavily infected rabbit sera and rSj338/26GST immunized rabbit sera by Western blotting. The inclusion bodies,the revivified protein and the purified protein from S 12 gel filtration were injected twice into rabbits to produce anti rSj338/26GST antibody which titer was then measured by dot ELISA. Balb/c mice were immunized with the inclusion bodies and the purified protein from S 12 gel filtration and challenged with S. japonicum cercariae to identify the protective immunity produced by rSj338/26GST. Results The inclusion bodies, the revivified protein and the purified protein from S 12 gel filtration could stimulate the rabbits to produce high level of anti rSj338/26GST antibodies and could be recognized by S. japonicum heavily infected rabbit sera and rSj338/26GST immunized rabbit sera. In Balb/c mice, the inclusion bodies could induce 27.8%( P

A 21-base, Plasmodium falciparum specific, enzyme-conjugated synthetic DNA probe (PFRl-AP) was used for the diagnosis of falciparum malaria. The blood samples (53 P. falciparum, 5 P. vivax, and 3 P. f and P.v mixed infection cases ) collected from Hainan province were tested. The samples of 32 college students were used for normal control. The probe proved to be specific and sensitive. 10pg of purified P.f DNA could be always detected, and there was no cross reaction with the purified DNA of human leukocytes. When testing Hainan blood specimens, PFRl-AP specifically detected P.f infections. In dot blot, when Nytran membrane with 50 microliters of treated blood samples being used, 39 out of 52 P.f specimens hybridized with this probe positively. When the volume of blotted sample was increased to one hundred microliters, the accumulative total positive rate rose up to 88.46%. The samples of P.v and normal control showed negative reaction with this probe.

In this study, the recombinant plasmid pPFl4 labeled with photobiotin was used as a probe to detect the patients infected with Plasmodium falciparum ( P. f.) by dothybridiza-tion. The results showed that out of 35 cases with P. f., 29 were positive, 5 were negative and one was doubtful. One patient with P.f. and P. vivax mixed infection showed positive result. The total positive rate was 83. 3% (30/36). 3 out of 33 normal human blood samples were positive, so the false positive rate was 9%. In addition, there was a correlation between the positive rate of detection and parasitaemia level. The detection sensitivity was 5 ?10-5.

Objective To obtain the specific IgE antibody related epitopes of Schistosoma japonicum from the phage display library. Methods Serum samples from 150 individials living in the epidemic regions of Schistosomiasis japonica were detected by ABC ELISA. 15 samples with high titer specific IgE antibodies were selected. Their pooled sera were absorbed with Protein G Sepharose beads to remove the IgG antibodies,then,it was used for immunoscreening of a phage display library of random peptide 12 mers. After 5 cycles of screening,DNA samples from 35 phage clones were sequenced. The phage clones with different inserted epitopes were identified immunologically. Results 4 independent phage clones of phage 3,phage 6,phage 8 and phage 15 were determined. Western blotting analysis showed that all of them could be recognized by specific IgE antibodies from the pooled sera. When they were used to immunize BALB/c mice,each clone could cause significant specific IgE antibody response. Conclusions The specific IgE antibody related epitopes of Schistosoma japonicum were screened successfully from the phage display library.

Objective To explore the effectiveness of ipsilateral autologous semitendinosus ten don graft in treatment of lateral ankle ligament injuries and ankle instability. Methods Two patients including one male (25 years old) and one female (17 years old) with chronic lateral instability of the ankle were enrolled in the study. Both patients had the history of repeated ankle sprain in supination position and had grade Ⅲ injury of the lateral ankle ligament according to the American College of Foot and Ankle Surgeons Grading System. Anterior drawer test and talar tilt test were all positive. The stress-inversion radiograph demonstrated the average inclination of the talus for 21 ° and the lateral radiograph demonstrated anterior dislocation of the talus. The chronic ankle instabilities in two patients were treated by using the ipsilateral autologous semitendinosus tendon graft to reconstruct the lateral ankle ligament. Results The two patients were followed up for mean eight months, which revealed that the active and passive range of motion was good, with no pain or swelling. The anterior drawer test and talar tilt test were all negative. The stress-inversion radiograph demonstrated that the average inclination of the talus was less than 5°, with no anterior dislocation of the talus. According to the Mazur grading system, the clinical outcome was excellent in one patient and good in one. The two patients were satisfied with the stability of the ankle. Conclusions ( 1 ) Lateral ankle ligament injury is a common cause of chronic ankle instability,even the ankle osteoarthritis. ( 2 ) Brostr(o)m method can attain satisfactory result for fresh lateral injury of the ankle, but not for the old injuries. ( 3 ) Ipsilateral autologous semitendinosus tendon graft is simpleand effective for treatment of lateral ankle ligament injuries and chronic ankle instability and may play an important role in the treatment of lateral ankle instability and prevention of the occurrence of ankle osteoarthritis.

Objective To construct and express human papillomavirus type 11(HPV11) E7 gene with recombinant adenovirus vectors. Methods HPV11 E7 gene was amplified by PCR and directionally cloned into vector pENTR-TOPO to form TOPO-E7 plasmid. E7 gene was transferred into the pAD/CMV/V5-DESTTM gateway vector by LR recombination reaction with pAD/CMV/V5-DESTTM gateway vectors and TOPO-E7 plasmid. The recombination vector was digested by Pac I enzyme and transfected into 293A cell by Lipofectamine method to obtain recombinant adenovirus vectors pAD-E7. Expression of E7 on HaCaT cells infected with pAD-E7 vectors was analyzed by confocal microscopy. Results The recombinant plasmid TOPO-E7 was identified and confirmed with enzyme digestion and sequencing. Recombinant adenovirus vectors pAD-E7 were generated efficiently with a titer of 1.4 ? 107 pfu/mL in transfected 293A cells. E7 protein could be identified in HaCaT cells with confocal microscope 48 h after infected with recombinant adenovirus vector. Conclusions The results indicate efficient expression of HPV11 E7 gene in eukaryotic cells by recombinant adenovirus mediated transfer, which facilitates further research of its function.

ngthen cellular immunity, especially Th1-type immune response to HPV11-E7 DNA vaccine in mice.