AIM: The purpose of this study was to establish a new strategy for constructing the mouse ? 2m gene targeting vector in order to increase the homologous recombination frequency in contrast with our previous one, which was successfully constructed in the normal way.METHODS: A 4.2 kb 3' arm and a 0.8 kb 5' arm were amplified by PCR from the mouse ? 2m-pSV2△HXgpt genomic clone. They included the start region and the three exons, which were separated into two parts from exons 2 (the main coding block) for the two arms——5' arm and 3' arm.RESULTS: The two fragments, in reverse orientation to the Neo gene, were cloned into pPNT respectively on either side of Neo. They were identified by PCR, restriction analysis and sequence analysis as well. CONCLUSION: The mouse ? 2m gene targenting vector has been cloned successfully.

Objective To investigate the relationship between C-reactive protein (CRP) and carotid intima-media thickness in pa-tients with impaired glucose tolerance (IGT). Methods Senan CRP was measured with immunoturhidimetry and the carotid intima-media thickness (CAIMT) was measured using color Doppler in 108 patients with impaired glucose tolerance (IGT) and 80 subjects with normal glucose tolerance(NGT).Then we observed all IGT patients for 3 years using prospective follow-up method, Oral Glucose tolerance test (OGGT) and every index were measured in follow-up 1.5 year and 3 year. Results 2 objects were lost to follow-up. IGT group showed a significant higher CAIMT and CRP compared with NGT group. After follow-up 1.5 year and 3 year, the patients with impaired glucose toler-ance (IGT) were divided into type 2 diabetes (T2DM) group and IGT group based on the level of blood glucose. Both T2DM group and IGT group showed a significant higher CAIMT and CRP, compared with NGT group. The level of serum CRP of T2DM group was higher than that of IGT group, and the level of serum CRP of IGT group was higher than that of NGT group. There were great differences between each group.Linear correlation showed that the level of blood glucose was positively correlated with CRP and CAIMT in T2DM group after follow-up 3 year. CAIMT was positively correlated with the level of blood glucose and CRP. Mulfivariant stepwise regression showed that CRP was signifi-canfly correlated with the level of blood glucose and CAIMT. Conclusion Inflammation played an important role in the development of dia-betes, and it had great vessels complication. The patients with impaired glucose tolerance, who have high level of CRP, were facilitated to be diabetes, and they were at risk of getting great vessels complication during the phase of impaired glucose tolerance. So it would be helpful to prevent IGT patients with high CRP or CAIMT with anti-inflammatory therapy.

BACKGROUNDStatin therapy has affected glucose homoeostasis of type 2 diabetes patients, which could be related with bile acids metabolism. Whether bile acid metabolism and the expression of farnesoid X receptor (FXR), liver X receptor-α (LXR-α) and sterol regulatory element-binding protein (Srebp)-1c is regulated by hyperglycemia, or whether simvastatin therapy led to higher glucose is related with down-regulated expression of FXR in diabetic rats remained unclear.

METHODSForty male Wistar rats were randomly divided into four groups: normal control rats, insulin resistance rats, diabetic model rats, and the late simvastatin induced diabetic rats. Normal control rats were fed with standard diet, others were fed with high-fat diet. Diabetic model rats were induced by a single intraperitoneal injection of streptozotocin (STZ). The late simvastatin induced diabetic rats started simvastatin administration after STZ induced diabetic model rats. Characteristics of fasting blood glucose (FPG), lipid files and total bile acids (TBAs) were measured and the oral glucose tolerance test (OGTT) was performed after overnight fasting at the eighth weekend. RNA and protein levels of FXR, LXR-α and Srebp-1c were tested by Western blotting and reverse transcription polymerase chain reaction (RT-PCR).

RESULTSThe insulin resistance rats showed higher glucose, lipid files and lower expression of FXR compared with normal control rats (P > 0.05). The diabetic model rats showed significantly higher glucose, lipid files, TBA and lower expression of FXR compared with insulin resistance rats (P < 0.05). The late simvastatin induced diabetic rats displayed higher glucose and TBA and lower expression of FXR compared with diabetic model rats (P < 0.05).

CONCLUSIONSChanges in bile acid homeostasis, including the alterations of bile acid levels and bile acid receptors, are either a cause or a consequence of the metabolic disturbances observed during diabetic models. Statin therapy induced hyperglycemia may be related with FXR, SHP, LXR-α and Srebp-1 pathways.

Animals ; Blood Glucose ; drug effects ; metabolism ; Diabetes Mellitus, Experimental ; drug therapy ; metabolism ; Glucose Tolerance Test ; Homeostasis ; drug effects ; Insulin Resistance ; physiology ; Liver X Receptors ; Male ; Orphan Nuclear Receptors ; metabolism ; Rats ; Rats, Wistar ; Receptors, Cytoplasmic and Nuclear ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Simvastatin ; therapeutic use ; Sterol Regulatory Element Binding Protein 1 ; metabolism