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 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 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 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.

[Objective] Based on the resting state functional magnetic resonance imaging to investigate the abnormal features of the functional connectivity of resting brain neural network in the patients with primary insomnia,by using voxel-wise whole-brain finctional networks analysis of degree centrality (DC) for imaging evidence of neural mechanisms underlying primary insomnia.[Methods] The resting state fMRI were performed in 59 PI patients and 47 age,education,and sex-matched normal healthy subjects.Analysis of DC map changes between the two patient groups and the control group were performed by two sample t test.(threshold at P ＜ 0.05).[Results] Compared with the control group,the patients with PI showed significantly reduced DC value in bilateral medial frontal gyrus (MFG),bilateral anterior cingulate gyrus (ACG),and right insula;and increased DC value in right middle temporal gyrus (MTG),and left cuneus,(CUN),P ＜ 0.05.[Conclusion]Changes of DC value occurred in some region of brain in the P[patient groups when compared with the control group.It was indicated that DC,as a novel resting-state fMRI parameter in the voxel-wise whole-brain functional networks,might be an appealing alternative approach for further study on pathologic and neuropsychological states of PI.

Objective To explore the effect of research practical teaching mode on the critical thinking ability and innovation ability of nursing in the course of community nursing teaching.Methods The research practical teaching was provided for 90 nursing students.They were investigated with the WatsonGlaser Critical Thinking Appraisal (WGCTA) and the test of self innovation level scale before and after teaching.Results After the teaching,the total score of critical thinking and the score of innovation ability among nursing students were significantly improved.the recognition hypothesis,deduction, explanation,evaluation discussed dimensions were statistically significant,but before and after teaching reasoningdimensions showed no significant difference.Conclusions Research practical teaching mode can improve critical thinking ability and innovation ability of nursing students in the course of community nursing teaching.