Objective:To explore the hepatocyte insulin sensitivity of intrauterine growth retardation ( IUGR) rats and establish an insulin resistance cell model in vitro.Methods: An IUGR animal model was established by protein malnutrition during the mother pregnancy .On 60 d and 90 d after birth , the offspring rats were fasted for 12 hours and then their angular vein blood was collected to measure the fasting plasma glucose and fasting serum insulin level , then the insulin resistance index ( HOMA-IR) and insulin sensitivity index ( ISI) were calculated .The insulin sensitivity was evaluated by HOMA-IR and ISI.Primary hepatocytes from each group were respectively isolated by two-step perfusion with collage-nase and were defined as normal hepatocytes group and IUGR hepatocytes group .The normal hepatocyte group was divided into two groups: control group and insulin induction group .Insulin induction group was established by primary cultures of normal hepatocyte incubated with varying dilutions of insulin . CCK-8 was used to detect the viability of the cultured hepatocytes .Glucose oxidase-peroxidase method kit was used to measure glucose consumption of the hepatocytes .Results:HOMA-IR was significantly higher in IUGR rats than in the normal rats at the age of 60 days ( t=-17 .02 , P<0 .05 ) and 90 days ( t=-12.52, P<0.05).ISI was significantly lower than in the normal rats aged 60 days (t=5.61, P<0.05) and 90 days (t=12.42, P<0.05).There were no significant differences in hepatocyte viability among the control group , IUGR group and insulin induction group after incubation of 48 h on day 60 (F=1.34, P=0.29) and day 90 (F=0.22, P=0.81).The glucose consumption of the IUGR group and insulin induction group were significantly decreased compared with the control group on day 60 ( F=9.28, P=0.002) and day 90 (F=56.60, P<0.001), while there was no significant difference be-tween the IUGR group and insulin induction group (P=0.08, P=0.10).Conclusion:The insulin sen-sitivity of hepatocytes of IUGR rats decreased from adolescence to adulthood .High-dilution insulin may induce insulin resistance cell model in vitro.

Objective To investigate the effect of liver phosphatidylinosital 3-kinase/protein kinase B (PI3-K/AKT) pathway on the decrease of insulin sensitivity in fetal growth restriction (FGR) rats.Methods Twenty pregnant female rats were randomly divided into two groups one day after conception:normal-protein group and low protein group (n=10,respectively).Rats in low-protein group was given low protein diet (8.00％ protein) during pregnancy to build FGR model,while normal-protein group was given normal protein diet (20.00％ protein).On day 3,7,14,30,60 and 90 after birth,fasting blood samples of 8 male FGR offsprings from low-protein group and 8 normal offsprings from normal-protein group were collected to measure fasting plasma glucose and insulin level.Then insulin resistance index and insulin sensitivity index were calculated to determine insulin sensitivity.On day 7,14,30,60 and 90 after birth,liver tissue of 8 male FGR and normal offsprings were collected,insulin receptor substrate 1,2 (IRS1/IRS2)and glucose transporter 4 (GLUT4) mRNA expression were measured by real-time fluorescence polymerase chain reaction and the protein expressions of IRS1,PI3-K (subunit p110β),and AKT and phosphorylated AKT (pAKT) were measured by Western blot.The relationships between the expression changes of key molecules of PI3-K/AKT pathway and insulin sensitivity were analyzed by correlation and multiple linear regression method.Results (1) Mean birth weight of baby rats in low-protein group was significantly lower than that of normal-protein group [(4.92±0.36) g vs (6.43±0.59) g,t=14.73,P＜0.05].The incidence of FGR in low-protein group was 88.2％ (97/110); and for male offsprings,it was 94.1 ％ (48/51).(2) Compared to normal offsprings,fasting plasma glucose levels of male FGR offsprings were significantly higher from the age of 60 days to 90 days.Insulin levels and insulin resistance index were significantly higher and insulin sensitivity index was lower from the age of 30 days to 90 days,P＜0.05 respectively.(3) Compared to normal offsprings,IRS1 (0.45 ± 0.02 vs 0.68± 0.03,t=16.633,P＜0.05) and IRS2 mRNA (0.34±0.10 vs 0.70±0.19,t=4.864,P＜0.05) expressions in FGR offsprings were lower from day 7 after birth to day 90 (0.48±0.03 vs 0.59±0.05,t=5.237,P＜0.05; 0.49±0.20 vs 0.70±0.11,t=2.253,P＜0.05).There were no differences in expressions of GIUT4 mRNA and AKT protein between two groups (P＞ 0.05).IRS1,PI3-K and pAKT protein expressions of FGR offsprings decreased significantly from day 14 (0.22±0.05 vs 0.52±0.11,t=7.024,P＜0.05; 0.46±0.03 vs 0.97±0.08,t=17.508,P＜0.05; 0.62±0.10 vs 0.89±0.08,t=6.100,P＜0.05) to day 90 (1.11±0.08 vs 1.32±0.14,t=3.714,P＜0.05; 0.63±0.07 vs 1.00±0.19,t=5.206,P＜0.05;0.28±0.03 vs 0.45±0.10,t=4.880,P＜0.05).(4) The pAKT protein expression level of FGR rats was positively correlated with insulin sensitivity index (r=0.704,P＜0.05) ; while negatively correlated to the level of fasting plasma glucose (r=-0.609,P＜0.05),fasting insulin (r=-0.561,P＜0.05) and insulin resistance index (r =0.577,P＜ 0.05).Conclusions The changes of some key molecules' expressions of PI3-K/AKT pathway in liver might be involved in the insulin resistance in FGR rats.

ObjectiveTo investigate the best pattern of the discussion on nursing cases of severe cardiac patients and improve the quality of the clinical nursing activity.Methods90 nurses from six cardiac units were divided into the control group and the experimental group with 45 nurses in each group. The experimental group adopted PBL teaching method, the control group adopted conventional teaching method. The discussion of severe cardiac patients was proceeded twice every month for one year.Results The experimental group was better than the control group in master and application of the basic cardiac knowledge, nursing quality of serious patients and effectiveness of nurse-patient communication.ConclusionsDiscussion with PBL teaching method is superior to the conventional method. This method can widen the nurses'thread, improve the nurses' ability to solve problem, so it is worthy of wide application.

Objective To investigate the MR imaging findings of acute experimental allergic encephalomyelitis(EAE) in correlation with pathology. Methods An EAE model was induced by intradermal inoculation with guinea pig CNS homogenate in 6 female Lewis rats.Another 6 rats served as control.The clinical presentation and body weight of the animals were recorded daily. Routine MRI,Gd-enhanced MRI were performed when EAE animals showed the initial symptoms. Uhrasmall superparamagnetic iron oxide(USPIO) colloid solution was also administrated intravenously and MRI was performed again after 24 hours. The brain was removed instantly after the second MR imaging. The pathological exams including HE staining,myelin sheath staining and prussian staining were performed.The imaging findings were observed in correlation with pathological results. Results The EAE rats showed decrease of body weight on the 6th to 7th day after inoculation,and the clinical symptoms appeared on the 10th to 11th day after inoculation.Routine MRI did not show any definite abnormalities.The Gd-enhanced MRI found the diffuse thickening and enhancement of brain meninges.The USPIO-enhanced MRI showedareas of low signal intensity at white matter of medulla oblongata on T2WI,and high signal intensity was observed at the corresponding area on T1 WI. Gradient T2 * WI found more foci of low signal intensity in eerebellar white matter besides the lesions in the brain stem.The range of abnormal signal intensity was larger in animal with higher clinical scores than that with lower score.There were no abnormal findings in control animaL The pathological exam found "perivascular cuff" in the brain white matter in EAE animals,some accompanied with adjacent demyelinatian. The prussian staining found blue particles within the cytoplasm of the macrophages around the lesion,which corresponded to the area of low signal intensity on T2WI.Conclusion USPIO-enhanced MRI could reveal acute EAE lesions which were not capable of being shown on routine MRI and Gd-enhanced MRI.It can image the macrophages around the lesions in vivo.USPIO is important for future research and application in MS patients.

Objective To explore the cataract surgery quality on blindness prevention and postoperative problems in village in short period. Design Population-based survey. Participants 251 cases(254 eyes) received operation and 131 cases(134 eyes)were surveyed 6-month postoperatively. Methods Patients were examined 6-month after the small incision extracapsular cataract extraction and intra ocular lens(IOL) implantation. Examinations were conducted by a special oculist including far vision, near vision, external in- spection, anterior segment, posterior segment, intraocular pressure. Main Outcome Measures Visual acuity, intraocular pressure, diopter, eye complications of surgery. Results Naked far vision of the operated eye more than or equal to 0.3 was 41.8%, naked far vi- sion of the eye more than or equal to 0.05 was 82.8%; corrected far vision more than or equal to 0.3 was 64.2%, corrected far vision more than or equal to 0.05 was 92.3%. Naked near vision more than or equal to 0.1 was 79.9%, corrected near vision more than or e- qual to 0.1 was 85.8%. The main postoperative complications were ametropia, posterior capsule opacification(PCO), deformed pupil, pupil displacement, pigments of IOL, eccentric IOL and intraocular hypertension. The chief reasons of eyes that could not be recovered were vitreous, retina or optic nerve diseases, the key factors that caused living vision less than 0.3 were ametropia, PCO, the disease of vitreous, retina and optic nerve. Conclusions The serious complications affecting the surgery result are limited in a low range. The most important factors of the eye corrected far vision less than 0.05 are the vitreous, retina and optic nerve diseases. In order to improve the visual sight, we should add equipment to calculate the IOL diopter accurately.

Objective To explore the value of fluorescence in situ hybridization (FISH) and multiplex RT-PCR in the detection of mixed lineage leukemia (MLL) gene rearrangement in acute leukemia (AL) patients. Methods Dual-color MLL probe, multiplex RT-PCR and R or G banding techniques were used to detect the MLL gene rearrangement in 189 cases of AL.Results MLL gene rearrangements were detected in 9 cases (5.03 ％) by FISH,and 16 cases (8.47 ％) by multiplex RT-PCR,including MLL/AF9,MLL/AF10,MLL/AF6, MLL/AF7, MLL/ELL, MLL/PTD. R or G banding techniques could find 11q23 in 5 out of 189 patients (2.65 ％). There was no statistic difference in the incidence of 6 common MLL gene rearrangements between ALL (73 cases) and AML patients (116 cases) (P ＞ 0.05).Conclusion Multiplex RT-PCR is a powerful technique in the detection of MLL gene rearrangement for tentatively diagnosed AL.It could not only confirm translocation detected by conventional cytogenetic method, but also detect MLL partial tandem duplication which could not been detected by cytogenetic examination or FISH. It plays an important role in guiding therapy and predicting prognosis for AL.

[Objective] To investigate combined application of multiplex reverse transcription-polymerase chain reaction(mRT-PCR)and karyotype analysis detect of clonal chromosomal aberrations in acute lymphoblasfic leukemia (ALL),and explore the expression of common fusion genes.Methods 189 ALL patients were examined by multiplex RT-PCR and R or G banding techniques.[Results]10 fusion genes were detected in 69 out of 189 ALL patients(36.5％),including E2A/PBX1,TEL/AML1,BCR/ABL,MLL/AF4,MLL/AF6、MLL/AF9,MLL/AF10,MLL/ELL,SIL/TAL1,TLS/ERG.R or G banding techniques could find chromosome structural and numeracy abnormalities in 86 out of 152 patients (56.6％) available for analysis.Combination of mRT-PCR and R or G banding could raise the rate of detecting clonal chromosomal abnormalities to 69.3％.Fusion genes were detected in 33 out of 90 (36.7％) patients with adult ALL and 36out of 99 (36.4 ％) patients with children ALL,there were 22 patients with positive BCR/ABL but no TEL/AML1 in adult ALL group,while there were 24 patients with positive TEL/AML.1 and 2 with positive BCR/ABL in children ALL group.There was significant statistical difference for the expression of RCR/ABL and TEL/AML1 between adult ALL and children ALL (P＜0.01),but no difference for MLL related fusion gene,E2A/PBX1,SIL/TAL1 and TLS/ERG(P＞0.05).BCR/ABL and TEL/AML1 fusion gene could be detected in 66 ALL patients with normal karyotype (36.3％).[Conclusion]There were different biological characteristics between adults and children with ALL.mRT-PCR technique can quickly screen chromosome structural aberrations in patients with newly diagnosed leukemia.It is useful in detection of fusion genes in ALL with normal karyotypes and it would refine the karyotype analysis and provide imrootramt prognosis-relevant information.

Objective To investigate the impact of polyclonal neural cell adhesion molecule antibody (P-NCAM-Ab) on the potency of botulinum toxin A (BTX-A).Methods Ninety male Sprague-Dawley rats were randomly divided into 3 equal groups:a normal control group,a BTX-A group and a P-NCAM-Ab group.The rats in the normal control group were injected with 100 μl of saline solution in their right gastrocnemius,while those in the BTX-A and P-NCAM-Ab groups were injected with 100 μl of BTX-A (0.5 U).In addition,the rats in the P-NCAM-Ab group were also injected with 100 μl of P-NCAM-Ab (the dosage was 20 U) at the same site on the 3rd day after the BTX-A injection.The rats' gastrocnemius muscle strength was evaluated with a self-made system for evaluating neuromuscular function before and after the toxin injection,on the 3rd day,as well as 1,2,4,6,8,10 and 12 weeks after the BTX-A injection.Any wet weight changes in the muscles were observed,and immunochemistry methods were employed to observe any structural changes in the motor endplates and nerve fibers at the different time points.Results After the saline injection,the average gastrocnemius muscle strength of the control group increased with time,while strength in the BTX-A and P-NCAM-Ab groups demonstrated a decrease in strength followed by a gradual increase.The average gastrocnemius muscle strength of the rats in the BTX-A and P-NCAM-Ab groups was significantly lower than that of the control group at all time points.Compared with the BTX-A group,the muscle strength of the P-NCAM-Ab group rats decreased further.Strength recovery in the BTX-A and P-NCAM-Ab groups was significantly slower than in the control group.The wet weight percentage in the BTX-A and P-NCAM-Ab groups at first decreased and then recovered with time.After the BTX-A injection,the average wet weight percentage of the P-NCAM-Ab group rats was significantly lower than that of the BTX-A group after 3 days,and 1,2 and 4 weeks.Karnovsky-Roots AchE staining showed that the motor endplates' color in the BTX-A and P-NCAM-Ab groups deepened gradually,though the color of the P-NCAM-Ab group was lighter than that of the BTX-A group at each time point.The mean optical density of the motor endplates' positive reaction area increased with time in both groups,but the P-NCAM-Ab group was lower than that of the BTX-A group at 1,2,4,8 and 12 weeks.Counting the nerve fibers dyed by gold chloride showed similar trends with both experimental groups significantly different from the control group.Conclusion P-NCAM-Ab can increase the potency of BTX-A and prolong its action.

Objective To explore the effect of methylation of peroxisome proliferator-activated receptor γ(PPARy) gene promoter in liver and its mRNA expression changes on decreasing of insulin sensitivity in fetal growth restriction (FGR) rats.Methods Twenty pregnant rats were randomly divided into two groups on their first day of pregnancy:normal-protein group (NP) and low-protein group (LP),ten in each.During pregnancy the LP group rats were fed with low-protein diet (8.00％ protein),while the NP group rats were fed with normal-protein diet (20.00％ protein).The offspring rats were fed with standard feed after 21 days of birth.Male offsprings in NP group were as control offsprings,and male FGR offsprings in LP group ware as FGR offsprings.At day 3,7,14,30,60 and 90,fasting blood of offsprings was collected to measure fasting plasma glucose (FPG) and fasting insulin(FINS).Then insulin resistance index of homeostasis model assessment (HOMA-IR) and insulin sensitivity index (ISI) were calculated to evaluate insulin sensitivity.At day 7 and 90,liver tissue of male offsprings was collected to extract DNA and total RNA.The methylation level of PPARγ gene promoter and its mRNA expression were detected by methylation specific-polymerase chain reaction (MS-PCR) and reverse transcription-RCR,respectively.The relationships between methylation of PPARγ gene promoter and mRNA expression and insulin sensitivity were analyzed by Pearson correlation and nonparametric test method.Results (1) The mean offspring birth-weight of LP group was (4.92±0.36) g,which was lower than that [(6.43±0.59) g] of control group (t=14.73,P＜0.05).In LP group,the incidence of FGR offspring was 88.2％ (97/110) and the FGR incidence of male ones was 94.1％ (48/51).(2) At day 90,compared with control offsprings,FPG [(8.95±1.83) mmol/L vs (6.21±1.14) mmol/L,t=-3.291,P＜0.05],FINS [(59.57±9.89) mU/Lvs (36.10±7.32) mU/L,t=-4.916,P＜0.05] and HOMA-IR (0.967±0.297 vs 0.410±0.135,t=-4.472,P＜0.05) of FGR offsprings were significantly higher; while ISI of FGR offspring was lower than that of control offsprings (-3.043±0.294 vs -2.172±0.354,t=4.774,P＜0.05).(3) There was no significant difference in methylation degree of PPARγ gene promoter in liver between FGR and control offsprings at day 7 (0/8 vs 2/8,Fisher exact test,P＞0.05).The methylation degree of PPARγ gene promoter in liver in FGR offsprings was significantly higher than that of control offsprings at day 90 (8/8 vs 2/8,Fisher exact test,P＜0.05).Compared with control offsprings,PPARγ gene mRNA expression level of FGR offsprings decreased significantly at day 90 (4.3.07±7.51 vs 146.72± 40.66,t=7.09,P＜0.05).mRNA expression of PPARγ gene was the lowest in exhaustive methylation offsprings (27.2± 1.6),and then in partial methylation ones (47.3±33.0),the highest in no methylation ones (144.6 ± 21.2) (P＜0.05).(4) The correlation analysis showed that PPARγ mRNA expression level negatively correlated to the level of FPG (r=-0.819),FINS (r=-0.906) and HOMA-IR (r=-0.860),P＜0.05 respectively; but positively correlated to ISI level (r=0.947,P＜0.05).Conclusions Hypermethylation in promoter region of PPARγ gene might inhibit gene transcription,and be involved in the occurrence of insulin resistance in FGR rats.

Objective HupA is one of the potential drugs which can be used to treat Alzheimer's disease(AD).The aim of this study was to explore the pharmacokinetics and biodistribution of HupA in vivo by using 11C-HupA.Methods A total of 25 SD rats were studied.They were divided into 5 groups (5 rats in each group).All had intravenous injection of 22 MBq(in0.2 ml)11C-HupA through tail vein.Dynamic im-aging Was acquired from 5 to 90 minutes after injection.Venous blood and organ activities were collected at 5,15,30,60.and 90 minutes after injection.Percentage activity of injected dose per gram of tissue(%ID/g)was calculated to characterize the biodistribution of tracer in different brain regions: frontal,apical, temporal,occipital,cerebellum,hippocampus,striatum,thalamencephalon, and brain stem, Variance analysis using SPSS 11.5 software was performed and compared among the study groups.Results 11C-HupA was character-istic for its quick clearance from blood,with half time T1/2 of (14.61±1.77) min,and clearance rate (CL)macokinetics of 11C-HupA in rats corresponded to a one-compartment model.with an activity curve(area 11C-HupA distribution in different brain regions,being greater in cerebral cortex,hippocampus,hypothala-mus and brain stem. Conclusions Pharmacokinetic study of 11C-HupA in brain was fast.convenient and showed high specificity and sensitivity.Its ability to quantitatively evaluate brain function and its character-istic distribution in mice provided some evidence for monitoring therapy in AD patients.