To establish a recombinase aided amplification ( RAA) assay for Plasmodium, we use the RAA assay to screen Plasmodium. Universal primers and probe were designed based on the specific fragment of the 18S rDNA gene of Plasmodium. We screened the primers and detected the specificity of the method. We screened four better amplification primers. RAA was performed at 37℃ in a short time (40 minutes) with good specificity. The RAA assay for Plasmodium detection appears to be applicable in ports.

Objective To investigate the significance of CerbB-2 oncogene in the occurrence and development of gastric adenocarcinoma. Methods By using immunohistochemistry and fluorescence in situ hybridization, 60 cases of gastric adenocarcinoma tissue, 60 cases of normal margin of mucosal tissue and 30 cases of normal gastric mucosa tissue were detected for the expression and amplification of CerbB-2. Results The positive rate of CerbB-2 expression in gastric adenocarcinoma tissues, normal margin of mucosal tissues and normal gastric mucosa tissues were 31.7 %, 8.33 % and 3.33 %; The positive rate of CerbB-2 amplification of these were 28.3 %, 5 % and 0. There were significant differences between gastric adenocarcinoma tissues and the others in the expression and amplification of CerbB-2. The expression and amplification of CerbB-2 in gastric adenocarcinoma tissues had no correlation with age,gender,tumor size or tumor differentiation degree of patients' (P >0.05), but were correlated with the extent of tumor invasion, lymph node metastasis, distant metastasis and clinical stage(P <0.05). Conclusion CerbB-2 is closely related to the occurrence and development of gastric adenocarcinoma, and can be considered as an important index for determining the biological behavior of gastric cancer.

Objective Using M2-Gi1α fusion protein expressed by baculovirus-Sf9 cell system to find the specific ligand for M2 receptor and detect the interaction of the two parts of the fusion protein.Methods The fused M2-Gi1α cDNAs were generated in a two-step PCR and then expressed in Sf9 cells.[3H]QNB and[35S]GTPγS binding experiments were employed to study the function of M2-Gi1α fusion protein.Results The expression level of M2-Gi1α fusion protein was 8.44±0.39 nmol·g-1 protein.The affinity of GDP to the Gi1α part changed under the affection of different ligands.The IC50 value in the appearance of acetylcholine,oxotremorine,arecoline,atropine,fangchinoline,levitimide were 21.35 μmol·L-1,23.86 μmol·L-1,11.91 μmol·L-1,0.13 μmol·L-1,1.05 μmol·L-1,1.75 μmol·L-1,and 2.5 μmol·L-1 when there was no ligand.Conclusion The M2-Gi1α fusion protein expressed in baculovirus-Sf9 cell system has pharmacological specificity for M2 receptor and the efficient coupling function between the two parts.The M2-Gi1αfusion protein is a helpful tool for detecting the new specific ligands of the M2 receptor.

Aim To generate m3AChR-G11 fusion protein in baculovirus-Sf9 cells and test the couping function,the interation and the influence factors be-tween m3AChR and G11 protein,as well as screen the specific ligands for m3AChR. Methods m3AChR-G11 fused DNA was generated through a two-step PCR and then expressed in Sf9 cells to produce fusion protein. The total concentration for membrane protein was de-tected by BCA method,[3H]QNB and [35 S]GTP?S binding experiment as perfomed to study the function of m3AChR-G11 fusion protein. Results The expression level of m3AChR-G11 was 7. 76 ? 10 -9 mol?g -1. The affinity of GDP to G11 partner changed in the presence of different muscarinic ligands. IC50 values of GDP in the presence of ACh,Pilo,CCh,MCN-A-343,Atro,4-Damp and Dafi were 82. 2,93. 70,12. 10,14. 30, 1. 93,1. 37,0. 72 ? 10 -6 mol ? L -1 respectively,and that in the absence of muscarinic was 1. 99 ? 10 -6 mol ?L-1.Concluslons The m3AChR-G11 fusion protein has the pharmacological specificity of m3 receptor and the efficient coupling interaction of the two partners. Affinity of GDP to ligand-bound fusion protein represents the species of muscarinic ligands. This is helpful in screening and detecting the new specific ligands to muscarinic receptors.