Objective To study the association between CYP2C9 gene polymorphism and warfarin maintenance dosage in anticoagulation therapy.Methods 200 Han patients admitted to our hospital for heart valve replacement were included in this study.CYP2C9 * 2,CYP2C9 * 3,CYP2C9 *c65 in CYP2C9 gene were sequenced using the CAPS technique and conventional DNA sequencing method.Dosages of warfarin used in patients carrying different genes were analyzed.Results No mutation of CYP2C9 * 2 but only one kind of allele C was detected in 200 patients.The genotype of CYP2C9 * 2 was C/C wild type.Allelic gene was detected at CYP2C9 * 3 A and C,with A/A wild type detected in 171 patients,A/C heterozygote mutation type detected in 18 patients,and C/C heterozygote mutation type detected in 11 patients respectively.The frequency of allelic genes A and B was 94.3 ％ and 5.7 ％ respectively.A significant difference was found between CYP2C9 * 3 mutation and warfarin dosage (P＜0.05).The dosage of warfarin reduced 18.46％ and 76.0％ respectively in patients carrying A/C heterozygote mutation type and in those carrying C/C heterozygote mutation type.Two kinds of allelic gene were detected at CYP2C9 * c65 G and C,with G/G wild type detected in 182 patients and G/C heterozygote mutation type detected in 18 patients respectively.No significant association was found in warfarin maintenance dosage for patients carrying G/G wild type and G/C heterozygote mutation type.Conclusion CYP2C9 gene polymorphism is associated with warfarin maintenance dosage in anticoagulation therapy.

BACKGROUND: Brain metabolic abnormality can be observed after cerebral ischemia.OBJECTIVE: To observe the changes of biochemical metabolism of rats with focal cerebral ischemia with 1H magnetic resonance spectroscopy (MRS) in order to reflect the metabolite abnormalities of rats during cerebral ischemic recovery phase.DESIGN: Randomized controlled study.SETTING: Departments of Neurology and department of Radiology of First Affiliated Hospital of Chongqing Medical University.MATERIALS: The experiment was conducted at the Radiological Department of First Affiliated Hospital of Chongqing Medical University from April to July 2004. Totally 24 adult Wistar rats with clean grade were randomly divided into three groups, namely: control group, sham operation group and cerebral ischemia group, with 8 rats in each group.METHODS: The focal cerebral ischemic model was established by occluding the right internal carotid artery in cerebral ischemia group and the filament were just inserted into the internal carotid artery not into the middle cerebral artery in sham operation group, nothing was done except for anesthetizing in the control group. 1H MRS was performed within the area of cerebral infarction and the homologous area of the contralateral hemisphere using 1.5T GE signa Highspeed MRI spectrometer in cerebral ischemic and shamed operation group at the following time point of 30 minutes, 1, 3, 6, 12, 24 hours, 3, 7, 15 days, 1 and 2 months after cerebral ischemia, and in the control group at the same time point.MAIN OUTCOME MEASURES: Changes of lactate, N-acetyl-aspartate (NAA), choline and creatine in infarct hemisphere and contralateral hemisphere.RESULTS: Totally 24 rats were selected in the study, but two died of anesthesia in sham operation group and four of serious brain edema in cerebral ischemia group, only 18 rats entered the final analysis with 8 in normal control group, 6 in sham operation group and 4 in cerebral isThe marked increase in NAA, choline and creatine was recognizable in bilateral spectra 30 minutes after cerebral ischemia and decreased to the period of 3-6 hours, but there was no significant difference in NAA, choline at 6 hours and reached the lowest level at 24 hours (45.21±0.37), choline (93.80±0.56) and creatine (69.33±0.44) at 3 days, and then NAA, choline and creatine gradually increased over time. The increase in NAA was especially obvious which increased by 2.5 times at 2 months than that at 24The earliest detection of lactate was at 10 minutes within the infarct. The lactate concentration elevated at 1 day and peaked at 12 hours, and a partial recovery of the reduction of lactate was seen at 3 days, and the lactate increase within the infarct region was significant compared with that of the homologous area (66.83±0.43,44.35±0.35, t=2.379, P ＜ 0.05). However, the elevation in lactate was no longer observed during the period of up to 2month follow-up.ischemia and reflect the metabolites changes in brain of rats after cerebral cerebral ischemic recovery phase objectively.

Objective To modify a classic two-vessel occlusion (2VO) modeling method in order to decrease the systematic errors in the behavioral experiments such as Morris water maze.Methods Thirty-two adult male Wistar rats were randomly allocated into classic 2VO model,modified model,sham operation and sham ligation groups (n =8 in each group).Only the bilateral common carotid arteries were ligated in the classic 2VO model group; the common carotid arteries were clipped intermittently,and the origins of pterygopalatine arteries of the internal carotid arteries were high selectively ligated in the modified model group; the common carotid arteries were only ligated intermittently in the sham ligation group; and only the common carotid arteries and the upper segment of pterygopalatine artery branches were separated in the sham operation group.The rat behavior was evaluated using the pupillary light reflex,Morris water maze and eight-arm radial maze.HE staining was used to observe the histological changes.Results The Morris water maze escape latency (F =72.169 - 163.102,all P ＜ 0.001) and the number of reference memory errors of eight-arm radial maze (F =33.515-74.726,all P ＜0.001) in the modified model and the classic 2VO model groups were longer and higher than those in the sham operation group.The pupillary light reflex of the rats was lost in the classic 2VO model group and the pupillary light reflex of the rats was normal in other groups.The reaching platform time in the classic 2VO model group was significantly longer than that in the modified model and sham operation groups (P ＜0.001).The percentage of target quadrant dwell time was also decreased significantly (at day 7 after procedure:F =13.770,P ＜0.001 ; at day 90 after procedure:F =14.780,P ＜0.001).HE staining showed pathological changes such as the cells decrease in hippocampal CA1 region and leukoaraiosis in the modified model and the classic 2VO model groups.In addition,there were more vacuole-like changes in the rat optic nerve region in the classic 2VO model group,while there were no such changes in the modified model group.Conclusions Establishing vascular dementia model with permanent occlusion of bilateral internal carotid arteries after intermittent occlusion of bilateral carotid arteries could avoid severe visual impairment in rats.In the Morris water maze and eight-arm maze test,the modified model rats showed significant decrease in learning and memory abilities and had hippocampal damage.

Objective To investigate the characteristics of traditional Chinese medical syndromes and syndrome patterns in diabetic nephropathy (DN) from delta regions of pearl river. Methods A cross-section and retrospective trial was carried out in 292 DN inpatients from four hospitals(the First Affiliated Hospital of Guangzhou University of TCM, Foshan Hospital of TCM, Shenzhen Hospital of TCM, and Zhongshan Hospital of TCM) from January, 2003 to December, 2005. Demographic information, past history, symptoms and laboratory parameters of the patients were collected.Results Of the enrolled 292 patients, there were 14 in the clinical stage Ⅱ, 42 in stage Ⅲ, 210 in stage Ⅳ, and 26 in stage Ⅴ. The manifestations of TCM syndrome were dominated as lassitude and weakness (69.9%), loss of energy (54.1%), dry mouth (49.3%), aching waist (42%), dry mouth and throat (38%), numbness of extremities (69.9%), dry eyes (33.2%) and dry stool (31.5%), but the signs of frequent sighing, night sweating and tidal fever were less found. For the deficiency syndrome patterns, there existed deficiency of kidney (37.7%), spleen (27.7%), heart (8.9%), liver (3.8%) and lung (2.1%) when differentiating the syndromes according the five zang-organs; there was deficiency of yin (32.5%), Qi (19.5%), blood (9.6%) and yang (8.2%) when differentiating the syndromes according to Qi, blood, yin and yang; there were complex deficiency syndromes such as deficiency of Qi and yin (40.4%), deficiency of spleen and kidney Qi (26.4%), deficiency of lung and kidney Qi(4.8%) and deficiency of yin and yang (3.8%). For the excess syndrome patterns, there were blood stasis syndrome (49.7%), damp-heat syndrome (33.2%), water-dampness syndrome (25%), and Qi stagnation syndrome (7.5%). Conclusion The syndrome patterns of DN present the following characteristics: leading deficiency syndromes being kidney deficiency and spleen deficiency when differentiating the syndromes according the five zang-organs, being yin deficiency and Qi deficiency when differentiating the syndromes according to Qi, blood, yin and yang, and complex deficiency syndromes being Qi-yin deficiency and spleen-kidney Qi deficiency. The leading excess syndromes are blood stasis syndrome and damp-heat syndrome. The manifestations of DN are characterized by the concurrence of deficiency and excess, cold and heat. The pathogenesis of DN is classified into deficiency in the root and excess in the branch. The deficiency syndrome mainly involves Qi and yin, and is located in spleen and kidney. The excess syndrome is characterized by blood stasis and damp-heat, and by water-dampness and dampness-turbidity in the late stage of DN. In one word, the pathogenesis of DN is characterized by deficiency of Qi and yin, and blood-stasis blocking kidney collaterals.

Objective To explore the effects of DPP-4 inhibitor sitagliptin on insulitis of NOD mice and the potential mechanism. Methods 223-month-old female NOD mice were randomly divided into Sitagliptin(Si-ta)group(n=11)gavaged with Sita(30 mg/Kg)daily for 12 weeks and control(Con)group with equal volume of normal saline(NS). The body weight,food-intake,water-intake and blood glucose were recorded weekly. Intraperi-toneal glucose tolerance test(IPGTT)was performed at the end of treatment and the blood sample was collected. Then mice were execute. The serum insulin level was measured by ELISA. Pancreas morphology and insulitis were evaluated by hematoxylin-eosin staining. The protein expression level of TLR4 ,MyD88 and NF-κB was analyzed by Western Blot. Results Compared with that of Con group ,the insulitis of Sita group was significantly alleviative(P < 0.05)and the serum insulin level was increased significantly(P < 0.05). The protein expression level of TLR4(P<0.05),MyD88(P<0.01)and NF-κB(P<0.05)in Sita group was significantly decreased. Conclusion Sitagliptin alleviates insulitis in NOD mice and increases the serum insulin level ,probably owing to the suppression of TLR4/MyD88/NF-κB signaling pathway.

Objective To explore the clinical efficacy of marginal liver from elderly donors in liver transplantation. Methods According to the inclusion and exclusion criteria, the clinical data of 127 donors and recipients were retrospectively analyzed. According to the age of donors, 127 donors were divided into the elderly group (n=27) and control group (n=100). The recovery of liver function, the occurrence of postoperative complications and survival rate of the recipients after transplantation were statistically analyzed between two groups. Results The incidence of primary nonfunction (PNF) and initial poor graft function (IPGF) did not significantly differ between the elderly and control groups (both P > 0.05). Within postoperative 2 weeks, the average levels of alanine aminotransferase (ALT) and serum total bilirubin (TB) of liver transplant recipients in the elderly group was not significantly different from those in the control group (both P > 0.05). There was no significant difference in the incidence of postoperative complications in the postoperative elderly group compared with the control group (all P > 0.05). The 1-and 3-year survival rates of the recipients in the elderly group were 84% and 78% respectively, which did not significantly differ from 89% and 79% in the control group (both P > 0.05). Conclusions Strict and sufficient quality evaluation and functional maintenance should be done for elderly donor livers. It can achieve good transplantation results by intraoperative fine operation, reducing bleeding and trauma, shortening the time of cold ischemia and operation, strengthening postoperative monitoring and implementing enhanced recovery after surgery.