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.
Animals ; Endothelial Cells ; metabolism ; Epidermal Growth Factor ; genetics ; metabolism ; Escherichia coli ; genetics ; metabolism ; Factor VII ; genetics ; metabolism ; Green Fluorescent Proteins ; genetics ; metabolism ; Rats ; Recombinant Fusion Proteins ; genetics ; isolation & purification ; metabolism ; Thromboplastin ; metabolism

INTRODUCTIONSingapore was one of 29 countries worldwide affected by severe acute respiratory syndrome (SARS) in 2003.

MATERIALS AND METHODSThere were 238 cases identified during the outbreak. We performed a retrospective analysis of the clinical and laboratory data of 234 patients admitted to Tan Tock Seng Hospital and Singapore General Hospital.

RESULTSThe mean age of patients was 21 years, 31.6% of patients were males and 41.8% were healthcare workers. At presentation, the common symptoms were fever, myalgia, cough and headache; rhinorrhoea was uncommon. On admission, 21% had leukopenia, 18% had thrombocytopaenia, 29% had hyponatraemia, 31% had hypokalaemia, 21% had transaminitis. Polymerase chain reaction (PCR) testing of respiratory and stool samples provided the best yield at the end of the first week of illness. Thirty-two patients were initially not recognised as probable SARS and were reclassified when the serology test results were available. The chief reasons for not identifying these patients early were persistently normal chest X-rays (68.8%), very mild presentation (43.8%) and the presence of a concomitant illness (12.5%). Overall, 12% of the patients were probable SARS with atypical presentations. Overall mortality was 11.8%.

CONCLUSIONPatients infected with the SARS coronavirus had a wide clinical presentation with non-specific symptoms.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Antibodies, Viral ; analysis ; Child ; Child, Preschool ; DNA, Viral ; analysis ; Diagnosis, Differential ; Female ; Humans ; Incidence ; Infant ; Male ; Middle Aged ; Retrospective Studies ; Reverse Transcriptase Polymerase Chain Reaction ; SARS Virus ; genetics ; immunology ; Severe Acute Respiratory Syndrome ; diagnosis ; epidemiology ; virology ; Severity of Illness Index ; Singapore ; epidemiology

Mycobacterium africanum is endemic to West Africa and is rare outside this region. Most of the people infected with M. africanum outside Africa are migrants from affected parts of Africa. We report a rare case of pulmonary tuberculosis (TB) secondary to M. africanum in a man in Brunei Darussalam who had lived and worked in Guinea, West Africa for 6 years more than 20 years ago. He had been well until December 2020, when he presented with a chronic cough and was diagnosed with coinfections of Klebsiella pneumoniae and M. africanum, and newly diagnosed diabetes mellitus. This case highlights an interesting manifestation of pulmonary TB secondary to M. africanum in a patient whose last exposure was 20 years ago, contributed to by development of diabetes mellitus.