Cryptosporidium infection in Nanjing City from 2015 to 2016 / 中国血吸虫病防治杂志

Objective To investigate the status of Cryptosporidium infection in the population in Nanjing City so as to pro-vide the evidence for the prevention and control of cryptosporidiosis. Methods A total of 100 fecal samples were collected from each of three districts(Liuhe,Qixia and Gaochun)and one hospital(Nanjing Zhongda Hospital)in 2015 and 2016 respective-ly. The fecal samples were detected for Cryptosporidium with microscopy(by using the gold amine phenol-modified acid-fast staining)and the positive samples were detected again for the molecular biology confirming by using the fluorescence quantita-tive PCR. Results During the two years,581 cases of normal population who lived in the city were surveyed and no Cryptospo-ridium infection was found. Among 202 cases of outpatients with chronic diarrhea,there were 9 Cryptosporidium positive cases with the microscope scanning method (4.46％),and among the 9 cases,7 cases showed obvious logarithmic amplification curves showing positive Cryptosporidium nucleic acid,but 2 cases without the obvious logarithmic amplification curves,and the Cryptosporidium nucleic acid positive rate was 3.47％. Conclusions Cryptosporidium infection is not found in the normal popu-lation of Nanjing City,but the Cryptosporidium infection is found in the chronic diarrhea patients. The results imply that we should strengthen the detection of Cryptosporidium in the chronic diarrhea patients,so as to provide the evidence for improving the diagnosis and treatment of cryptosporidiosis.

Expression of fusion protein encoding EGFP-EGF1 of rat coagulation factor VII and its binding function / 中国实验血液学杂志

This study was aimed at clarification of the function of EGF(1) segment in rat coagulation factor VII with tissue factor (TF) by means of the expression of the fusion protein of EGFP-EGF(1). The DNA fragment encoding EGF(1) was amplified from a rat liver tissue by RT-PCR, and then inserted in an EGFP-procaryotic expression vector to construct the recombinant plasmid pET28a-EGFP-EGF(1) which was introduced into the competent cells of E.coli BL21, then an engineering bacteria strain was obtained which was induced by IPTG to express the fusion protein of EGFP-EGF(1). The fusion protein was purified by chromatography on Ni column, and then acted on the rat hemangioendotheliocytes stimulated with LPS to express TF; the binding of the fusion protein to the hemangioendotheliocytes was detected by means of fluorescence microscopy and flow cytometry. The results indicated that EGFP-EGF(1) was highly expressed in the engineering E.coli strain, and successfully purified, and its molecular mass was confirmed as 36 kD by SDS-PAGE. Fluorescence microscopy and flow cytometry had shown that this fusion protein can bind with the TF on the hemangioendotheliocytes. It is concluded that the EGF(1) region of rat coagulation factor can mediate the specific binding of FVII with TF, so as to lay partly the basis for molecular targeting anti-thrombotic therapy.