In recent years, more and more biological materials are used in bone defect repair. The ideal materials should not only have good biocompatibility, but also have the right space structure, surface morphology and the physical and chemical properties, to adapt the growth of osteoblasts. Calcium phosphate cement was used more widely among biological materials in bone defect repair with its good biocompatibility, degradability, osteoconductivity, injection, plasticity and other advantages. This paper introduced the experimental research and progress of the calcium phosphate cement application in bone defect repair.

Rhoptry proteins are the major virulence factors of Toxoplasma gondii. They locate in different parts of the host cells and can affect the membrane cytoskeleton structure and active factors of the host cells so as to block the cell intrinsic de-fense mechanisms of the host and let T. gondii invade parasitize and proliferate in the host successfully. The function and ac-tion mode of rhoptry proteins reflect the pathogenic mechanism of T. gondii which holds great significance to looking for toxo-plasmosis drug targets and developing molecule vaccines. This paper reviews the research progess of the interaction between rhoptry proteins of T. gondii and host cells.

Toxoplasma gondii rhoptry proteins (ROPs) include dozens of proteins secreted by tachyzoites ,these ROPs are known to be critical for several processes such as invasion ,parasitophorous vacuole establishment ,proliferation ,egress and getting out of host immune protection .ROPs belong to the protein kinase family and have classic kinase domain ,however ,most of the members are designated as pseudokinases due to the lack of critical catalytic residues that are normally required for phos-phorylation of host signal molecules .Recent studies indicated that pseudokinases also have important functions ,such as regula-ting kinase activity by allosteric and promoting the interactions between proteins as scaffolds .In this paper ,we summarize the pseudokinases structural characteristics and some pseudokinases functions ,for the study of toxop lasma gondii and host interac-tion mechanisms ,molecular vaccines screening and drugs design .

Objective To observe the therapeutic efficacy of acupuncture at the interiorly-exteriorly related meridians plus rehabilitation training in treating post-stroke shoulder-hand syndrome. Methods Sixty eligible patients with post-stroke shoulder-hand syndrome were randomized into a treatment group and a control group, 30 in each group. The treatment group was intervened by acupuncture at the interiorly-exteriorly related meridians plus rehabilitation training, while the control group was by rehabilitation training alone. After 2 treatment courses, the Fugl-Meyer Assessment (FMA), visual analogue scale (VAS) and edema score were observed and compared. Results The FMA, VAS, and edema scores were significantly changed in both groups after intervention (P＜0.01). There were significant differences in comparing the FMA, VAS, and edema scores between the two groups after intervention (P＜0.05). The total effective rate was 83.3%in the treatment group versus 76.7%in the control group, and the difference was statistically significant (P＜0.05). Conclusion Acupuncture at the interiorly-exteriorly related meridians plus rehabilitation is an effective method in treating post-stroke shoulder-hand syndrome.

Objective To develop a rapid molecular biological method for detection of the asymptomatic infection of Leish?mania. Methods Two pairs of primers named RV1?RV2 and K13A?K13B were selected to be the fast diagnosis primers since they were designed according to the conserved region of Leishmania kinetoplast DNA(kDNA)minicircles. The PCR amplifica?tion products of Leishmania donovani promastigote from Shandong Province were sequenced to compare their conservatism. The method was applied to detect 105 venous blood samples from healthy home canine and 7 venous blood samples from home canine suffered from Kala?azar in Heishui County of Sichuan Province,and 75 venous blood samples from susceptible population(no leishmaniasis symptoms)and 7 venous blood samples from patients in Xinjiang Kashi area in order to verify the feasibility and accuracy of the method. Results The size of PCR products was consistent with the expected fragments with high conservative among Leishmania species. The positive rates of 105 home canine samples and 75 susceptible population samples were 37.14%(39/105)and 82.67%(62/75)rspectively,and the positive rates of home canine suffered from Kala?azar and patients were all 100%(7/7). Conclusion This rapid diagnosis method is suitable for detection of asymptomatic infection of Leishmania in Kala?azar endemic areas of China with high sensitive and specific,thus it has bright perspective to be used.

Objective To investigate the feasibility and effect of intentional occlusion of the left subclavian artery (LSA) in endovascular aortic repair EVAR of thoracic aortic disease.Methods The cases,who needed occlusion of the LSA in EVAR of thoracic aortic disease,were examined cerebral circulation,carotid artery,vertebral basilar artery and circle of Willis.If the vertebral artery had a good blood pathway,and carotid artery and circle of Willis had no stenotic,we occluded the LSA directly in EVAR,and observed the brain and upper limb ischemia complications postoperation.Results Among 40 cases who were occluded the LSA directly in EVAR,28 cases were successful with no neurological complications and left upper limb ischemia symptoms postoperation,12 patients with mild symptoms of subclavian steal syndrome and neurdogical complications,but none required and secondary surgical intervention.Conclusions Prior to intentional LSA occlusion,attention must be paid to potential supraaortic variants and pathologies.Only in this way,it was safe and effective to occlude the LSA in EVAR of thoracic aortic disease.

Objective To subclone express and identify the immune mapped protein 1 IMP1 which encodes a surface an?tigen of Toxoplasma gondii. Methods The cDNA of T. gondii RH strain was synthesized by reverse transcription PCR the IMP1 open reading frame ORF was amplified by PCR using the T. gondii RH strain cDNA as template the PCR products were identified by TA?cloning and sequencing then the IMP1 ORF was subcloned into the NdeⅠand Xho I sites of the vector pET28b and the positive recombinant pET28b?IMP1 was identified by double?digesting and sequencing. The protein of 6 × His tagged IMP1 was inducibly expressed in E. coli strain BL21 DE3 with isopropylβ?D?1?thiogalactopyranoside IPTG and the induction time concentration of IPTG and temperature gradients to optimize protein expression conditions were determined. After the cells carried IMP1 were induced by the optimized conditions and harvested the resulting bacteria were suspended in resuspension buffer and lysed by sonication and the supernatants were loaded onto the Ni2+Chelating Sepharose Fast Flow col?umn for affinity chromatography of the N?terminal 6 × His tagged IMP1 protein. Finally the fusion IMP1 proteins were identified by Western blotting. Results The ORF sequence of IMP1 was successfully subcloned from the cDNA of Toxoplasma Gondii RH strain and the amplified product was sequenced and identified based on which the IMP1 ORF gene was inserted into the prokaryotic expression vector pET28b and the recombinant pET28b?IMP1 was constructed successfully. The double?digesting and sequencing results indicated the validity of the recombinant vector. And the optimized conditions for the expression of IMP 1 was determined namely 0.3 mmol/L IPTG induction for 9 h at 20℃. Furthermore IMP1 protein was expressed solubly and che?lated on Ni2+sepharose beads with high affinity thus this protein could be purified efficiently by affinity chromatography. The pure fusion protein was confirmed with fine immunocompetence by SDS?PAGE and Western blotting. Conclusions IMP1 pro? tein can be high efficiently expressed by the E. coli prokaryotic expression systems the protein of IMP1 is soluble and has stable characters. The study may lay a useful foundation for the following works including in vivo expression of IMP1 crystal structure study of IMP1 and anti?toxoplasmosis subunit vaccine development.

Objective To provide vessel anatomical materials guidance for endovascular aortic repair,the branches artery of thoracic aortic was studied by CT angiography (CTA).Methods From January 2008 to February 2012,739 adult cases' CTA data were collected,all cases performing thoracic CTA in Wuhan General Hospital of Guangzhou Command.We measured the diameter and/or leugth of the ascending aorta,aortic arch and branches artery of aortic arch,and made an analysis.Results The aortic arch includes standard and variant types.Standard type is common,which accounted for 91.1％ of the total number,while variant type accounted for 8.9％.In the standard aortic arch of patients,the diameter of aortic arch above the opening of coronary artery (CA) was (35.7 ±4.3) mm,the diameter of ascending aortic arch at the opening of brachiocephalic trunk (BCT) was (33.6 ±4.2) mm,the diameter of aortic arch between the BCT and the left common carotid artery (LCCA) was (29.4 ± 5.7) mm,the diameter of aortic arch between the LCCA and the left subclavian artery (LSA) was (27.6 ± 4.2) mm,the diameter of descending aortic at the opening of the LSA was (25.4 ± 4.5) mm,the diameter of the head BCT from aortic arch was (12.9 ±0.9) mm,the diameter of the head LCCA from aortic arch was (8.5 ± 0.7) mm,the diameter of the head LSA from aortic arch was (10.4 ± 1.1) mm,the length of aorta between the CA and the BCT was (53.3 ±12.5) mm,the length of aortic between the BCT and the LCCA was (4.7 ± 1.5) mm,the length of aortic between the LCCA and the LSA was (7.9 ± 2.6) mm,the length between the opening of BCT and the right subclavian artery (RSA) was (41.1 ± 8.2) mm,the length between the opening of LSA and the opening of left vertebral artery was (38.5 ±5.7) mm,the angle between the horizontal of BCT and the LCCA and the sagittal plane was (71.2 ± 7.2) °,the angle between the plane of LCCA and the LSA and the plane of sagittal was (31.1 ± 2.9)°.Conclusions The CT data of the thoracic aorta can be used as reference for production of stents and guide releasing the stents in endovascular repair.

Objectives To summarize the clinical characteristics of imported falciparum malaria patients and the treatment so as to provide the evidences for improving the diagnosis and treatment of the disease. Methods A total of 138 imported falci?parum malaria patients who received the treatment in Shandong Institute of Parasitic Diseases from January 2007 to February 2013 were adopted as the observation subjects and their clinical data were collected and analyzed. Results All the 138 pa?tients were back from African countries. The main manifestations were fever headache asthenia and hepatosplenomegaly and most of them were with decreased RBC PLT levels and increased LDH levels and 36.96%of them were misdiagnosed as respiratory diseases nephritis hepatitis and so on. Through antimalarial treatment of artemether or artesunate or dihydroartemis?inin and primaquine or dihydroartemisinin and piperaquine and symptomatic treatment the short?term and long?term cure rates were 98.55%and 94.93%respectively with 1 case unrecovered and 1 died. Conclusions Artemisinins are still the most effective antimalarial drugs for falciparum malaria. However some patients recrudesce as the Plasmodium in their body is resis?tant or insensitive to these drugs. We should pay more attention to the antimalarial and symptomatic treatments in the early stage of severe malaria so as to improve the cure rate.

Objective To establish the primary cat intestinal epithelial cells(IECs)culture methods and construct the cD-NA library for the following yeast two-hybrid experiment,so as to screen the virulence interaction factors among the final host. Methods The primary cat IECs were cultured by the tissue cultivation and combined digestion with collagenase XI and dispase I separately. Then the cat IECs cultured was identified with the morphological observation and cyto-keratin detection ,by using goat anti-cyto-keratin monoclonal antibodies. The mRNA of cat IECs was isolated and used as the template to synthesize the first strand cDNA by SMARTTM technology,and then the double-strand cDNAs were acquired by LD-PCR,which were subsequently cloned into the plasmid PGADT7-Rec to construct yeast two-hybrid cDNA library in the yeast strain Y187 by homologous recom-bination. Matchmaker?Insert Check PCR was used to detect the size distribution of cDNA fragments after the capacity calcula-tion of the cDNA library. Results The comparison of the two cultivation methods indicated that the combined digestion of colla-genase XI and dispase I was more effective than the tissue cultivation. The cat IECs system of continuous culture was established and the cat IECs with high purity were harvested for constructing the yeast two-hybrid cDNA library. The library contained 1.1× 106 independent clones. The titer was 2.8 × 109 cfu/ml. The size of inserted fragments was among 0.5-2.0 kb. Conclusion The yeast two-hybrid cDNA library of cat IECs meets the requirements of further screen research,and this study lays the foundation of screening the Toxoplasma gondii virulence interaction factors among the cDNA libraries of its final hosts.