Objective To explore the effects of high glucose on glucose transport activity, and phosphorylationandexpressionofinsulinsignalingproteins in primary cultured rat adipocytes. Methods Isolated rat adipocytes were cultured at different glucose concentrations (5, 10, 15, 25 mmol/L) for 24 h. Then the glucose uptake, the phosphorylations of insulin receptor (IR), insulin receptor substrate (IRS) 1 and 2 and protein kinase B (PKB) as well as the protein expressions of IRS1, IRS2, p85 subunit of phosphatitylinositol 3 kinase (p85) and PKB were measured. Results These adipocytes treated with different high glucose showed the impairment of the basal and insulin induced increase in glucose uptake and significant decrease of IR, IRS1 and PKB phosphorylations as well as IRS1 protein expression, but up regulation of IRS2 protein expression. p85 and PKB contents and IRS2 phosphorylation were unaffected. Conclusion The exposure to high glucose inhibits glucose uptake and induces insulin resistance in adipocyte. The mechanism may be involved in affecting the multiple step phosphorylations and the expressions of insulin signaling proteins.

Objective To investigate the expression of forkhead box C2 (FOXC2),Vimentin and E-cadherin protein in colorectal adenocarcinoma and the relationship of FOXC2,Vimentin and E-cadherin with the clinicopathological parameters,and the role of epithelial-mesenchymal transition (EMT) in colorec tal adenocarcinoma and the correlation between FOXC2 and EMT.Methods The expressions of FOXC2,Vimentin and E-cadherin in 102 cases of colorectal adenocarcinoma tissues and 30 cases of adjacent noncancerous tissues were detected by immunohistochemical method,and the correlation between FOXC2 and EMT and the clinicopathological parameters of colorectal adenocarcinoma were analyzed.Results The positive rates of FOXC2,Vimentin and E-cadherin in colorectal adenocarcinoma tissues were 53.9％,40.2％ and 26.5％,respectively.The positive rates of FOXC2,Vimentin and E-cadherin in adjacent noncancerous tissues were 6.7％,0 and 63.3％.Compared to paracancerous tissue,the expression of FOXC2 and Vimentin in colorectal adenocarcinoma was significantly increased,while the expression of E-cadherin was significantly decreased.The positive rates of FOXC2 and Vimentin in lymph node metastasis group were 79.1％ and 67.4％,respectively,which were significantly higher than those without lymph node metastasis (35.6％,P ＜0.01;20.3％,P ＜0.01).The positive rate of E-cadherin in lymph node metastasis group was 16.3％,which was significantly lower than that without lymph node metastasis (33.9％,P ＜ 0.01).The positive rates of FOXC2 and Vimentin in T1 +T2 phase were 41.7％ and 25.0％,respectively,which were significantly lower than those in T3 + T4 stage (83.3％,P ＜0.01;76.7％,P ＜0.01).The positive rate of Ecadherin in T1 + T2 stage group was 33.3％,which was significantly higher than that in T3 + T4 stage (10.0％,P ＜ 0.01).There was a positive correlation between FOXC2 and Vimentin in colorectal adenocarcinoma (P ＜0.01),and negatively correlated with E-cadherin expression (P ＜ 0.01).Conclusions EMT may promote the development and metastasis of colorectal adenocarcinoma,FOXC2 may be involved in colorectal cancer EMT and through the EMT to promote the malignant process of colorectal adenocarcinoma.

With the increasingly tense doctor-patient relationship,the practice teaching of laboratory medicine has obviously been affected.For example,the practice teachings systems do not adapt to the updated situations,and practice teaching has been simplified and communication training ignored.In order to deal with these problems and ensure the goals of practice teaching,various measures have been taken in training hospitals.Firstly,the practice system has been further improved and implemented effectively by strengthening mobilization and adopting qualification licensing before practice and interaction practice between teachers and students and regular communication between school and training hospital during practice and strict assessment after practice.Secondly,medical ethics educations and laboratory biological safety have been strengthened.Thirdly,the modem automation and classic morphology should be integrated.Finally,the ability of communication between medical laboratory and doctor and patients should be cultivated,and so on.These measures are to ensure the effect of the practice teaching and cultivate excellent students.

To study effects of arachidonic acid (AA) on the growth and differentiation of rat adipocytes, a cells culture system of rat primary preadipocytes was established. The cells treated by different concentration of AA supplemented based on DMEM medium containing 10% fetal bovine serum. Cell proliferation was measured by trypan blue exclusion and methyl thiazolyl tetrazolium (MTT) assay method. Hoechst33342 fluorescence staining observed AA induced morphological changes. Oil Red O staining extraction assay assess the degree of adipogenesis and differentiation, and cyclooxygenases-2(COX-2) mRNA were detected by reverse transcriptase-polymerase chain reaction (RT-PCR). The results showed that rat preadipocytes treated with 120 μmol/L AA for 24-72 hours remarkably promoted the cells proliferation compared with control, 160 μmol/L AA treated for 48 hours could induce apoptosis of preadipocytes. 40, 80 μmol/L AA decreased the fat content in cells at 72 hours, and 40 μmol/L AA significantly up-regulated the expression of COX-2 mRNA at 24 hours. This results indicate that AA regulate adipocytes proliferation and differentiation depended on treatment time and concentration. 40-80 μmol/L AA maybe useful to control body fat, which may be associated with the increase of COX-2 mRNA.

Objective To investigate the long-term effects of intrauterine mild hyperglycemia exposure and postnatal high fat diet on the body weight and metabolism of offspring through a pregnant rat model of intrauterine mild hyperglycemia.Methods Twenty-one pregnant Wistar rats were randomly divided into intrauterine hyperglycemia group and control group.Twenty percent streptozotocin (STZ,25 mg/kg)was given to rats of intrauterine hyperglycemia group by a single intraperitoneal injection to induce intrauterine mild hyperglycemia; control group rats received an equal volume of citric acid-sodium citrate buffer.Off springs were divided into 4 groups:exposed to intrauterine hyperglycemia and fed with normal diet group(group DN)or high fat diet group (group DF) ; exposed to intrauterine euglycemia and fed with normal diet group(group CN)or high fat diet group(group CF).The blood glucose levels of pregnant rats in two groups and body weights of offsprings in four groups were recorded.At the age of 28 weeks,the mesenteric fat amount,epididymal amount,perirenal fat amount,total triglyceride (TG)and high density 1 ipoprotein-cholestrol(HDL-C) were measured in all four groups.Results (1) The average blood glucose level of intrauterine hyperglycemia group[(16.6 ± 3.4) mmol/L] was significantly higher than that of the control group [(5.8 ± 1.1) mmol/L,P ＜ 0.01].(2) On the birth day,3 weeks and 4 weeks,the body weight of group DN [(7.4 ± 0.6),(44.1 ± 5.9),(79.6 ± 7.4) g] and group DF [(7.4 ± 0.2),(43.9 ±6.9),(76.1 ± 5.8) g] were remarkably increased compared with group CN [(6.6 ± 0.5),(35.6 ± 4.4),(71.5±6.8) g,P＜0.05]; but the body weight in group CF [(6.7 ±0.5),(33.0 ±6.5),(66.1 ±10.2) g] had no statistical difference compared with group CN(P ＞ 0.05).(3)From then on,the bodyweights of the offsprings in four groups presented an increasing trend,but there was no statistical difference until 28 weeks(P ＞0.05).(4) The perirenal fat amount of group DN,group CF and group DF [(13.8 ±3.3),(14.3 ± 3.2),(18.4 ± 1.3) g] were remarkably increased compared with group CN [(9.7 ± 3.5) g,P ＜ 0.05] ; the epididymal fat amount of group CF and group DF were also significantly increased compared to group CN(P ＜ 0.05) ; the mesenteric fat amount in four groups had no statistical difference (P ＞ 0.05).(5) The TG level of group DN,group CF and group DF[(0.52 ±0.14),(0.52 ±0.09),(0.54 ±0.17)mmol/L] were significantly higher compared to group CN [(0.41 ± 0.09) mmol/L,P ＜ 0.05],but there was no statistical difference within the first three groups (P ＞ 0.05) ; the HDL-C level in four groups had no statistical difference(P ＞ 0.05).Conclusions In intrauterine mild hyperglycemia environment,there were some evidently metabolic changes observed in the offspring,including body weight increasing on birth day and early postnatal period,visceral fat amount increasing and lipid metabolism disorders,which could be aggravated by postnatal high fat diet.

Objective To investigate the relationship between insulin resistance index (HOMA-IR) and the clinical pathology fea-tures in chronic hepatitis B(CHB) patients. Methods ALT and hepatitis B virus loading level and HOMA-IR were measured in 80 CHB pa-tients. Correlation analysis was performed with pathological features of liver tissue. Result Insulin resistance(IR) was much more obvious in patients with severer degree of liver inflammation and fibrosis . HOMA-IR and the insulin level of CHB patients in G3 grade(15.24±7.41,3.51 ± 1.91) and S3 stage (16.65±6. 46,3.79 ± 1.50) were higher than those in G2 grade (10.33 ± 6.28,2.25 ± 1.39) and S2 stage(9.06 ± 4.61,2.02 ± 1.11). The HOMA-IR was positive correlated with the level of age and ALT. Conclusion The development of IR in CHB patients was correlated with age and the degree of liver inflammation and fibrosis.

Objective Study the effects of early overfeeding in the adult offspring of mother with severely hyperglycaemia in pregnancy to islet development and insulin resistance. Methods Thirty healthy female Wistar rats were mated with 10 male Wistar rats and the morning on which sperm were found in three different visual fields of the vaginal smear was designated pregnancy day 1. The pregnant rats were intraperitoneally administered with Streptozotocin (STZ, 50 mg/L) on 5th day of pregnancy, and blood glucose exceeded 20 mmol/L to induce severely gestational diabetes mellitus (SDM) model. The pregnant Wistar rats were assigned to two experimental groups: SDM (n = 16) and control (n = 8). Litter size reduction in the lactation period induced early postnatal overfeeding model. Offspring were divided into three groups according to the level of blood glucose in pregnancy and feeding patterns in lactation: ( 1 ) control group (CG):euglycemia in pregnancy, eight pups in lactation;(2) severely gestational diabetes mellitusnormal feeding (SDM-N):severely gestational diabetes mellitus, eight pups in lactation; (3) severely gestational diabetes mellitus-overfeeding (SDM-O): severely gestational diabetes mellitus, four pups lactation. At the end of the lactation period, all pups were fed standard laboratory chow adlibitum until the date of the experiments. Offspring body weight was measured weekly after ablactation. Serum insulin was measured by enzyme-linked immunosorbent assay (ELISA) and pancreatic islet morphology was analyzed by immunohistochemistry (IHC) in all three groups at 26 weeks of age. Results (1) Blood glucose of pregnant Wistar rats: SDM (28.3 ±5.1 ) mmol/L was statistically higher than control (6.3 ± 1.4) mmol/L ( P ＜ 0.05 ). (2) Growth rates of body weight in 3 - 7 weeks and 3 - 9 weeks: SDM-N: (4. 6 ± 1.3) % and (6.8±2.5)%, SDM-O:(3.2±0.7)% and(4.6±1.2)%,CG:(2.9 ±0.6)% and(4.1 ±0.8)%.The growth rates of body weight in SDM-N and SDM-O were both significantly higher than those in CG ( P ＜0.05 ). (3) Body weight at 26 weeks: CG: (486 ± 132) g, SDM-N: ( 387 ± 115 ) g, SDM-O: ( 382 ± 122) g.There was no statistical difference among the three groups (P ＞0.05). (4) Fasting plasma glucose ( FPG),fasting insulin (FINS), homeostasis model of insulin resistance (HOMA-IR) and insulin sensitivity index (ISI): at 26 weeks, the SDM-offspring has nomal FPG, but more insulin was needed to keep it normal. The insulin level of SDM-O[ ( 12.6 ± 3.3) mU/L ] was statistically higher than those of SDM-N [ ( 10.9 ± 3.3 )mU/L] and CG [ ( 8.6 ± 0.8 ) mU/L ] ( P ＜ 0.05 ). The ISI of SDM-O ( 0.020 ± 0.006 ) was significantly smaller than its HOMA-IR(2.40 ±0.62,P ＜0.05). (5)The morphological change of pancreatic islet: The islets of CG and SDM-N were round or ellipse and have clear boundary between endocrine and exocrine parts and the β cells distributed equally. However, SDM-O islets were not of uniform size and most of islets were hyperplasia and hypertrophy. (6) Relative β cell area of pancreas, β-cell area and islet size: SDM-O:(1.81 ±0.31)%, (57.1 ±3.2)% and(39 067 ±3308)μm2; SDM-N:(1.34 ±0.43)%,(60.9 ±0.6)% and ( 30 570 ± 4824 ) μm2; CG: ( 1.11 ± 0.26 )%, ( 63.7 ± 2.7 )% and ( 26 443 ± 4431 ) μm2.SDM-O has significantly increasing β-cell mass, hypertrophic islet size and slightly decreasing β-cell percentage compared with other two groups (P ＜ 0.05 ). Conclusions The exposure of severely hyperglycemia in pregnancy induces low weight infant and postnatal catch-up growth leading to the possibility of insulin resislance (IR) in adult and early postnatal overfeeding will accelerate such course. Islet morphology of SDM-N has no significant change, indicating that maternal diabetes mainly affected β-cell function but not islet morphological features. SDM overfeeding results in early impairment of islet morphology and function, indicating that the compensation ability of islets has already been impaired and the risk of further development of impaired glucose tolerance (IGT) and diabetes. In conclusion, the nutritonal environment in early life ( duration of pregnancy and lactation) participate in the metabolic programming in adulthood.

Objectives To investigate the clinical significance of degree Ⅲ meconium-stained amniotic fluid in term pregnancy.Methods Data of 857 full-term gravidas underwent cesarean section for suspected fetal distress in Peking University First Hospital from October 1,2012 to September 30,2014 were retrospectively analyzed.All patients were divided into three groups according to the diagnosis of suspected fetal distress:meconium-stained amniotic fluid group (Group 1,n=172),fetal heart abnormality group (Group 2,n=623) and meconium-stained amniotic fluid in combination with fetal heart abnormality group (Group 3,n=62).General information and fetal prognosis of the three groups were analyzed.Factors that might affect the prognosis of newborns were analyzed between the infants with or without acidosis,asphyxia or aspiration pneumonia.Analysis of variance,independent-samples t test,Chi-square test and rank-sum test were used as statistical methods.Results There were no significant differences in maternal age,gestational age at delivery,parity,nuchal cord loop(s),pH value of umbilical arterial blood and gender of newborns among the three groups (all P＞0.05).The neonatal Apgar score,neonatal asphyxia rate and rate of admitting into pediatric ward within 24 hours after birth in Group 1 were similar to those of Group 2 and 3 [1 min Apgar score:9.9±0.7 vs 9.8±0.7 and 9.8±0.7,F=0.322;5 min Apgar score:10.0±0.3 vs 10.0±0.2 and 10.0±0.0,F=0.517;neonatal asphyxia rate:1.7％ (3/172) vs 2.1％ (13/623) and 1.6％ (1/62),x2=0.129;rate of admitting into pediatric ward:16.3％ (28/172) vs 14.3％ (89/623) and 11.3％ (7/62),x2-0.978] (all P＞0.05).The incidence of neonatal aspiration pneumonia in Group 1 was higher than that of Group 2 and 3 [4.7％ (8/172) vs 3.2％ (2/62) and 1.0％ (6/623)],but the differences had no statistical significance (F=10.680,P=0.050).Conclusions Both degree Ⅲ meconium-stained amniotic fluid and abnormal fetal heart rate have the same indicating significance and should be treated actively to lower the risk of poor prognosis of the newborns.

Objective To explore the preventive effect and mechanism of pioglitazone on nonalcoholic fatty liver disease in rats.Methods Thirty six male Sprague-Dawley ( SD) rats were randomly divided into normal control group , high-fat diet group , and pioglitazone intervention group , with 12 rats in each group .Normal control group fed with standard diet for eight weeks , and the rest two groups fed with high fat diet .Pioglitazone intervention group fed with pioglitazone in the four to eight weeks , the rest two groups in the same period fed with distilled water .Fasting serum alanine aminotransferase , aspartate aminotransferase , total cholesterol , triglyc-erides, fasting glucose , and fasting blood insulin levels were determined .The fasting insulin resistance index was calculated .Routine histological features of hepatic section were observed by hematoxylin eosin (HE) staining.The shape change of Kupffer cells in the liv-er was detected .The levels of tumor necrosis factor ( TNF)-αand nitric oxide ( NO) secreted by Kupffer cells were measured .Results The fasting insulin resistance index , triglycerides , and total cholesterol in rats of high-fat diet group were significantly higher than in normal control group ( P <0.05 ) .Histopathological examination showed hepatocellular macrove sicular steatosis , lobular inflammatory cell infiltration, and inflammatory cell infiltration .Hepatic Kupffer cell morphology change , its produced TNF, NO and liver tissue pa-thology changes were positively correlated ( P <0.05 ) .The fasting insulin resistance index , triglycerides , total cholesterol was signifi-cantly lower in rats of pioglitazone intervention group than the high-fat diet group ( P <0.05 ) .The liver tissue structure was obviously improved pioglitazone intervention group rats , morphology and function of liver Kupffer cells were normal .Conclusions Pioglitazone can prevent the nonalcoholic fatty liver disease induced by high fat diet .The mechanism may be related to improve insulin resistance , reduce blood fat , and regulate function of Kupffer cells .

Objective To investigate the effect of solution-focused therapy on the care burden and negative emotions of the primary caregivers with elderly patients with hip fractures. Methods One hundred and sixty-eight elderly patients with hip fractures and caregivers were divided into intervention group and control group with 84 pairs by random digits table method, and the two groups were given continual nursing care. At the same time, the primary caregivers of intervention group accepted the solution-focused therapy. Caregiver Burden Inventory (CBI), Self-rating Anxiety Scale (SAS), and Self-rating Depression Scale (SDS) were used as the evaluation indexes. Results Before intervention, the score of CBI, SAS,SDS was (42.07 ± 4.24), (50.75 ± 11.21), (51.59 ± 13.18) points in control group and (42.10 ± 3.96), (50.48 ± 12.59), (52.96 ± 13.51) points in intervention group, and there was no significant differences between two groups (P>0.05). After intervention, the score of CBI, SAS, SDS was (40.49±3.17), (50.64±11.33), (50.14±10.25) points in control group and (28.95±2.87), (36.73±9.45), (41.60±8.23) points in intervention group, and there was significant differences between two groups (t=5.42, 8.73, 7.45, all P<0.05). Conclusions Solution-focused therapy can effectively reduce the care burden of the primary caregivers of elderly patients with hip fractures and alleviate their anxiety and depression.