Expression and purification of bioactive high-purity human midkine in Escherichia coli.

Midkine is a heparin-binding growth factor, which plays important roles in the regulation of cell growth and differentiation. The non-tagged recombinant human midkine (rhMK) is therefore required to facilitate its functional studies of this important growth factor. In the present work, rhMK was expressed in Escherichia coli (E. coli) BL21 (DE3). The expression of midkine was efficiently induced by isopropyl-beta-D-thiogalactopyranoside (IPTG). After sonication, midkine was recovered in an insoluble form, and was dissolved in guanidine hydrochloride buffer. Renaturation of the denatured protein was carried out in the defined protein refolding buffer, and the refolded protein was purified using S-Sepharose ion-exchange chromatography. The final preparation of the rhMK was greater than 98% pure as measured by sodium dodecylsulfate-polyacrylamid gel electrophoresis (SDS-PAGE) and reverse phase high performance liquid chromatography (RP-HPLC). The purified rhMK enhanced the proliferation of NIH3T3 cells.

Investigation on road traffic safety of the mountain areas in southwest China based on Wulong County, Chongqing Municipality.

OBJECTIVE: To study the present situation, characteristics, and main problems of the traffic safety in the mountain areas of Southwest China. METHODS: A study was done on the traffic status of Wulong County, Chongqing Municipality, by means of document review, field investigation, and inquiry. RESULTS: The traffic crashes and deaths in Wulong County were increasing since 1999. From 1997 to 2001, there occurred 126 severe and very severe traffic crashes resulting in 175 deaths and 411 serious injuries, which accounted for 9.4% of total crashes, 96.1% of total deaths, and 48.4% of total serious injuries, respectively. The severe and very severe traffic crashes were highest in 2000. One part for 83 km in length accounting for 3.0% of No. 319 National Highway was responsible for 54.5% of very severe traffic crashes, which closely related to conditions of the road itself and its environment and defective execution of traffic laws. CONCLUSIONS: Wulong County, one of the mountain counties in Southwest China, is now facing a complex and serious road traffic situation and serious traffic safety, with a high incidence of severe and very severe road traffic crashes. The crashes closely relate to imperfect road design, defective execution of traffic laws, imperfect management, and environmental factors.