Objective To investigate the safety and accuracy of diagnosing alpha-thalassemia with extraembryonic coelomic cells. Methods Coelocentesis was performed before artificial abortion to collect extraembryonic coelmic fluid in 40 women with singleton pregnancy during 6-10 gestational weeks. The villi was gathered after suction. PCR technique was used to amplify alpha-thalassemia gene in both extraembryonic coelomic cells and villi and concordant rate of two different kinds of samples were compared. Results The genotypes of alpha-thalassemia were successfully amplified in 37 cases of coelomic cells and all were concordant with the results of villi. Conclusion Extraembryonic coelomic cells can be used in early prenatal diagnosis of alpha-thalassemia by PCR technique and is practicable.

Based on the background,conception and the core ideas of the evidence-based medicine,the construction and application of domestic clinical cases database was discussed.Problems concerning statistics,management and application in scientific research encountered in the database devel opment were presented,and suggestions were proposed.

Objective To investigate the correlations between HLAⅡgene polymorphism and the development of Mycoplasma pneumoniae pneumonia ( MPP ) in children and to identify the susceptibility genes and protective genes for MPP for further elucidating the pathogenesis of MPP and providing the guid-ance for researches on gene therapy for MPP. Methods Genotypes of HLAⅡgene ( HLA-DQA1 and HLA-DRB1) in 60 children with MPP and 30 healthy children were detected by using sequence specific primer polymerase chain reaction ( SSP-PCR) . The haplotype frequencies, linkage disequilibrium and correlations with MPP were analyzed by using Arlequin software and Chi-square test. Results The frequencies of HLA-DQA1*0201/*0301 in children with MPP (35. 83%/30. 00%) were higher than those in healthy children (16. 67%/8. 33%) (χ2=12. 139, P<0. 05, OR=5. 059;χ2=15. 142, P<0. 05, OR=7. 500). However, the frequency of HLA-DQA1*0401 in children with MPP decreased to 12. 50% as compared with 40. 00%in healthy children (χ2=24. 638, P<0. 05, OR=0. 083). The frequencies of HLA-DRB1*07/*15 in children with MPP increased to 38. 33%/30. 00% as compared with 20. 00%/16. 67% in healthy children (χ2=11. 735, P<0. 05, OR=4. 929; χ2=5. 692, P<0. 05, OR=3. 000). But the frequency of HLA-DRB1*11 dropped to 15. 00% as compared with 43. 33% in healthy children (χ2=19. 448, P<0. 05, OR=0. 087). Conclusion HLA-DQA1*0201, HLA-DQA1*0301, HLA-DRB1*07 and HLA-DRB1*15 might be the susceptibility genes for MPP in children, while HLA-DQA1*0401 and HLA-DRB1*11 were probably associated with the resistance to MPP. No extensive linkage disequilibrium was found between HLA-DQA1 and HLA-DRB1.

BACKGROUND: Many study results proved that electroacupuncture can cause change of endogenous opioid peptide system.OBJECTIVE: To observe the change of blood sugar and blood lipoprotein (a) in rats with diabetes mellitus after electroacupuncture therapy and probe into the effect of opioid peptide system in the treatment for diabetes with electroacupuncture.DESIGN: Randomized controlled trial with experimental animals as the subjects.SETTING: The experiment was conducted in the Department of Pathophysiology, Medical College of Shandong University.MATERIALS: The experiment was completed in the Department of Pathophysiology, Medical College of Shandong University from July 2004 to December 2004. Totally 140 male Wistar rats were involved.METHODS: Ten Wistar rats were collected randomly as the blank control group. The other 130 rats were given 20 g/L streptozotocin 50 mg/kg intraperitoneally and the rats in blank control group were given the same volume of saline intraperitoneally. Three weeks later, the tails were cut off and the blood sugar was measured 2 hours after food intake. The rats with obvious rapid sugar tolerance were given foods with high calories for 28 days as the model rats. There were 64 rats were successful models and they were classified into 3 groups randomly, namely model control group( n = 20), electroacupuncture therapy(n=22) and electroacupuncture with naloxone groups( n = 22) . The blood sugar and blood lipoprotein(a) were detected after treatment for the diabetic rats with streptozotocin.MAIN OUTCOME MEASURES: Blood sugar level and blood lipoprotein(a) of each rat after the final therapy.RESULTS: Totally 74 rats entered the result analysis. Electroacupuncture therapy could reduce the blood sugar and blood lipoprotein(a) of diabetes mellitus rats, while rats with naloxone injection before therapy were insensitive to electroacupuncture. [The blood sugar value were(13.91 ± 3.79),( 10.33 ± 2.36), (12.96 ± 2.23)mmol/L in the model control group, electroacupuncture therapy group, electroacupuncture with naloxone group respectively; while the level of blood lipoprotein(a) were(13.75 ±2.69),( 10.81 ± 2.96), (13.01 ± 3.23)mg/L respectively].CONCLUSION: Electroacupuncture therapy can reduce the level of blood sugar and the risk of atherosclerosis in rats with diabetes mellitus. Although the mechanism can not be explained through the meridian theory, according to the experimental result that blood sugar and blood lipoprotein(a) were no longer intervened by electroacupuncture therapy after injecting naloxone, it is presumed that electroacupuncture therapy can exert its intervention effect on blood sugar and blood lipoprotein(a)through opiate system.

Objective To explore the feasibility of coelocentesis in prenatal diagnosis.Methods Coelocentesis were applied on 58 women scheduled for at 6～12 weeks of gestation,and fetal heart rate before the procedure and 1,5,30 minutes afterwards and hemorrhage of amnionic cyst was observed.Y gene of sex-determining region was detected by polymerase chain reaction,and compared with chorionic villi sample.Results The total achievement ratio of coelocentesis was 96.6% of all cases in five minutes.The coelomic fluid was successfully aspirated at first attempt in 93.2% at 7～10 week's gestation,25.0% at 6～7weeks and 50.0% at 10 ~ 12 weeks.There was no significant difference in FHR between before coelocentesis and afterwards (P＞0.05).The detection accurate rate of SRY was 91.1%,all female fetus were matched with those observation by chorionic villi sampling,and only 5 male fetus were not matched,the accurate rate was 80.8%.Conclusion Coclocentesis has the advantage of simple procedure,safe and high feasibility.The optimal time of coclocentesis is in the 7th to 10th week of gestation.

Objective To evaluate the effect of angiotensin Ⅱ ( Ang Ⅱ ),angiotensin- (1-7) [ Ang- ( 1-7 ) ],and co-action of Ang Ⅱ and Ang-( 1-7 ) on β cell insulin signaling pathway.Methods Mouse pancreatic β cell line NIT-1 was incubated with( 1 )0,10-7,10-6,10-s,10-4 mol/L concentrations of Ang Ⅱ for 24 h ; ( 2 )0,10-7,10-6,10 -5,10-4 mol/L concentrations of Ang- ( 1-7 ) for 24 h; ( 3 ) co-administration of Ang Ⅱ and Ang- ( 1-7 ) was divided into control,10-5mol/L Ang Ⅱ,10-6mol/L Ang-( 1-7 ),10-5mol/L Ang Ⅱ + 10-6mol/L Ang-( 1-7 ) group.Tyrosine phosphorylation of insulin receptor β subunit(IR-β-Tyr) and serine phophorylation of protein kinase B(Akt-Ser) were detected by Western blot.ResultsInsulin-stimulated IR-β-Tyr and Akt-Ser phosphorylation was significantly decreased in Ang Ⅱ 10-5 and 10-4 mol/L group; no significant changes in insulin-stimulated IR-β-Tyr and Akt-Ser phosphorylation were detected between Ang-( 1-7 ) treatment groups and control; Ang-( 1-7 ) blocked the inhibitory effect of Ang Ⅱ on Akt-Ser phosphorylation,yet exerted no effect on Ang Ⅱ-induced IR-β-Tyr phosphorylation inhibition.Conclusion Ang Ⅱ significantly inhibits insulin signaling pathway in β cell; Ang-( 1-7 ) reverts the inhibitory effect of Ang Ⅱ on insulin-stimulated Akt-Ser phosphorylation in β cell.

Objective To construct a rat model of nonalcoholic fatty liver disease (NAFLD) by feeding rats fat-rich diet and analyze the effect of insulin resistance (IR)in the development of NAFLD. Methods Male SD rats were randomly divided into normal diet group (NG, n =24) and fat-rich diet group (FG, n =24). At the end of feeding for2 weeks, 4 weeks, 6 weeks or 8weeks, 6 rats in NG and FG were randomly took out. Their weight were recorded, then the serum fasting blood sugar, fasting insulin, triglyceride, total cholesterol, ala-nine aminotransferase and aspartate aminotransferase were measured, and the fasting insulin resistance index and liver index (liver weight (g)/body weight(g) × 100%)was calculated. Then liver tissues were homogenized, and muleic dialdehyde and superoxide dismutase were determined. The hepatic steatosis in all rats was assessed according to the results under light microscope. Results The body weight of rats in NG increased faster than those in FG after six weeks. The liver index of rats in FG was markedly higher than that in NG since the second weekend. The rats in FG began to have hepatocyte steatosis from the second weekend, had insulin resistance, hyperlipidemia, dysfunction of liver and lipid peroxide of liver from the fourth weekend, suffered mild fatty liver from the sixth weekend, and developed to moderate fatty liv-er from the eighth weekend. Conclusions NAFLD with IR model was successfully developed by feeding SD rats with an improved rich-fat diet for 6 weeks. IR may play an important role in the development of NAFLD.

Objective To evaluate the right heart function with echocardiography after right ventricle-pulmonary artery (RV-PA) anastomasis for right ventricle outflow (RVOT) reconstruction in patients with different types of pulmonary atresia and ventricle septal defect(PA/VSD).Methods From Nov 2002 to Aug 2013,31 patients with PA/VSD had undergone right ventricle-pulmonary anastomasis to reconstruct RVOT for radical or palliative repair.Related echocardiography indexs including strain/rate etc.were used to evaluate the right heart function and the progress of the right heart valves regurgitation.Results There were 3 early hospital deaths.No later death during follow-up.The echocardiography suggested the pulmonary artery and tricuspid regurgitation were more serious,however,the right heart function was relatively fine.The regurgitation of tricuspid valve was positive correlation with duration of follow-up (P =0.016).Conclusion The right heart function in follow-up keeps relatively well,and tricuspid valve regurgitation needs a long-term follow-up.

Objective To establish a mouse model of kidney-yin and kidney?yang deficiency after oral administra?tion of hydrocortisone, and to explore the related evaluation factors. Methods The model was established by oral adminis?tration of hydrocortisone to induce kidney?yin and kidney?yang deficiency in mice. The survival and body weight of the mice were observed. The serum content of adrenal cortical hormone ( ACTH) , cortisol ( Cor. ) in the hypothalamic?pituitary?ad?renal (HPA) axis, and thyroid stimulating hormone (TSH), triiodothyronine (T3), thyrox (T4) in the hypothalamic?pi?tuitary?thyroid (HPT) axis, follicle?stimulating hormone (FSH), estradiol (E2), testosterone (T) in the hypothalamic?pituitary?gonadal ( HPG) axis were determined by radioimmunoassay. Results The body weight of kidney?yin and kidney?yang mice were decreased, the serum ACTH, Cor, TSH, T3, T4 contents were decreased, the serum FSH, E2, T contents were increased in the kidney?yang deficiency model mce ( P<0. 01 ) , and those parameters in the kidney?yin deficiency model mice were changed in opposite direction. Conclusions It is found that the hormone levels of ACTH, Cor, TSH, T3, T4, FSH, E2 and T in kidney deficiency mice are changed, and cortisol can be used as an important index to evaluate the model of kidney deficiency induced by glucocorticoid.

Objective To compare the two different ways of right ventricular outflow tract(RVOT) reconstruction at repair of pulmonary atresia with ventricular septal defect,the direct RV-PA anastomosis and pericardial conduit to find the better way.Methods From Jun.2002 to Oct.2012,66 patients underwent pulmonary atresia with ventricular septal defect repair in our hospital,age at operation from 14 days to 272 months.Patients were divided into 2 groups according to the way of RVOT reconstruction.Group 1:31 of them,using direct RV-PA anastomasis,Group 2:35 of them,using pericardial conduit.Paired t test was used to evaluate the growth of pulmonary arteries.Chi-square test and Kaplan-Meier were used to calculate the postoperative mortality,reopemtion situation and survival time.Results There are 3 early hospital death in group 1 (9.7 ％),and 5 in group 2(14.3％),P =0.71.There is a significant difference between the two groups in restenosis rate of the RV-PA anastomasis and autologous pericardial conduit with pulmonary branch artery(Group 1:22.2％,Group 2:55.6％,P =0.01).The diameters of RV-PA anastomasis and the pulmonary artery branches in follow-up were significantly lager than the earlier diameters(P ＜ 0.05) in group 1.There is no growth on diameters of the pericardial conduit and pulmonary branches except the right pulmonary artery in follow-up in group 2.There is no significant difference between the two groups in later survival(P =0.30).Conclusion Both the direct anastomasis of RV-PA and pericardial conduit are available for RVOT reconstruction in pulmonary atresia with ventricular setal defect repair.There is lower incidence of RVOT and pulmonary stenosis and anastomosis absolutely has the ability for later growth in the former.