Objective To study the effect of exogenous prostaglandin E 1 (PGE 1) on the superoxide dismutase(SOD) and nitric oxide(NO) levels in brain tissue of neonatal rats with hypoxic-ischemic brain damage(HIBD).Methods Sixty 7-day old newborn Wistar rats to establish HIBD models,intraperitoneally and subcutaneous injected PGE 1 and TMP,then the rats were killed after hypo- xia and ischemia for 48 hours.Take cerebral cortex of arteria carotis ligation side and made them into homogenate to detect SOD and NO levels in brain tissue.Results SOD level in HIBD group was lower,and NO level was higher than those of normal group(P

Objective To investigate whether the spores from mushroom antigen can cause the allergic pneumonia and manufacture allergic animal model in the C57BL/_6 mouse.Methods Aged 6 weeks old,weight 25-30 g C57BL/_6 mice were collected.In the mouse tail injection compound of spore antigen and the Freud′s adjuvant.Then pours into through the trachea the antigen once a week.The mice were divided into 4 groups.Group A was the normal mouse,Group B was given Freud′s adjuvant(the same method)to determine whether there was affect to the mouse.Group C and D were injected spore antigen 2 and 4 times.When the antigen sensitization finished 1 week later group C and D were completely divided 2 groups,among them one group was inhalation 1.5% spore antigen and induce the acute response.Six hours later the bronchoalveolar lavage fluid(BALF) was collected to observe cell change,and excise the lung tissue to manufacture the pathology specimen,another group had not been induced the acute response and collect the BALF and to exsise the lung tissue directly.Group B were inhalted saline later to collect the BALF and the lung tissue.In the mouse blood serun,enzyme linked immunosorbent assay(ELISA) was used to mensurate antigen specific IgG.Results In group C and D,antigen specific IgG significantly inhanced than that in group A and B(all P

Objective To explore the relationship between sex hormone-binding globulin (SHBG) of gestational diabetes mellitus ( GDM ) pregnant women with well-controlled glucose and pregnancy outcomes. Methods Two hundred and fifty-one GDM pregnant women of 24 - 28 weeks in Shengjing Hospital of China Medical University were recruited from Mar. 2005 to Mar. 2010. Two hundred and sixteen cases of GDM with well-controlled glucose were defined as glycemic satisfied group, and they were treated by diet therapy ( 169 cases) or insulin therapy (47 cases) . Thirty-five cases with unsatisfied glucose were defined as glycemic unsatisfied group. One hundred and ninety-two healthy pregnant women of 24 - 28 weeks were defined as healthy control group. Serum SHBG and homeostasis model analysis of insulin resistance ( HOMA-IR) at 24 - 28 weeks and above 36 weeks were measured. GDM was diagnosed bytwo-step method according to the National Diabetes Data Group ( NDDG) criteria. The pregnancy outcomes and complications of the three groups were recorded. Results ( 1 ) Comparison of pregnancy outcomes and complications: glycemic satisfied group was less likely to develop hypertensive disorders in pregnancy ( 10. 6% ) , premature birth(8. 3% ) ,large for gestational age ( LGA) (8. 8% ) , neonatal asphyxia(3. 7% ) and neonatal hypoglycemia ( 2. 3% ) compared to glycemic unsatisfied group ( 42. 9% , 34. 3% , 31. 4% , 22. 9% and 11. 4% ,respectively). And the difference was statistically significant (P ＜0. 05 or P ＜0. 01). There was no significant difference for incidence of polyhydramnios, pueperal infection, postpartum hemorrhage, neonatal hyperbilirubinemia between the two groups ( P＞ 0. 05 ) . When compared to healthy control group(7. 3% ,2. 1% ,4. 2% ,2. 1% and 1. 6% ) ,no significant difference was found for incidence of premature birth( 8. 3% ) , pueperal infection ( 3. 2% ) , postpartum hemorrhage (5. 1% ) , neonatal asphyxia (3. 7% )and neonatal hypoglycemia(2. 3% ,P ＞0. 05). (2) Comparison of results of 24 - 28 weeks and above 36 weeks: serum SHBG of glycemic satisfied group [( 384 ± 88 ) , (457 ± 48 ) nmol/L]was significantly higher than that of glycemic unsatisfied group[(313 ±45) ,(401 ±73) nmol/L];HOMA-IR of glycemic satisfied group (5. 3 ±1.1,5.5 ±1.1) was significantly lower than that of glycemic unsatisfied group (7. 0 ± 1. 3 ,7. 6 ± 1. 7 ; P ＜ 0. 01). Serum SHBG of glycemic satisfied group was significantly lower than that of healthy control group [( 492 ± 95 ) , (565 ± 40 ) nmol/L]; and HOMA-IR of glycemic satisfied group(5. 3 ± 1. 1,5. 5 ± 1. 1) was significantly higher than that of healthy control group (3. 6 ±0. 6,3. 9 ± 0. 5 ;P ＜ 0. 01 ) . FPG of glycemic satisfied group [( 5. 84 ± 0. 28 ) , ( 5. 16 ± 0. 13 ) mmol/L]was significantly lower than that of glycemic unsatisfied group [(6. 13 ± 0. 16 ) , ( 5. 68 ± 1. 14) mmol/L; P ＜ 0. 01]. FINS of glycemic satisfied group [( 20. 4 ± 2. 1 ) , ( 24. 1 ± 4. 2 ) mmol/L]was significantly lower than that of glycemic unsatisfied group [(24. 7 ± 4. 5 ) , ( 29. 9 ± 2. 7 ) mmol/L; P ＜ 0. 01]. ( 3 ) Correlation analysis. Between 24 - 28 weeks, SHBG was negatively correlated with HOMA-IR in the three groups ( r = -0. 952, P ＜0. 01) ; and SHBG was negatively correlated with HOMA-IR in glycemic satisfied group ( r = -0. 903, P ＜0. 01). Conclusions Well-controlled glucose can not completely improve maternal and fetal outcomes of GDM pregnant women. High insulin resistance and low serum SHBG can influence pregnancy outcomes.

Objective To observe the effect of circadian rhythm on hypnotic median effective dose( ED50) of ketamine. Methods Sixty mice were randomly divided into 4 groups which had 15 mice in each group. They were intraperitoneally injected with ketamine at different times of 2 Am,8 Am,2 Pm and 8 Pm, respectively. Righting reflex was recorded and the value of ED50 was measured with sequential experimental method. Results The hypnotic ED50 of ketamine at 2 Am was(54.57?0.82) mg/kg, with 95% confidence limit of ED50 38.06-78.22 mg/kg;ED50 was(49. 27?0. 12) mg/kg at 8 Am, with 95% confidence limit of ED50 40. 21-60. 37 mg/kg;ED50 at 2 Pm was (42.28?0.21) mg/kg, with 95% confidence limit 37.35 - 47 83 mg/kg;and ED50 at 8 Pm was(57.42?0.14) mg/kg, with 95% confidence limit 37.51-73 72 mg/kg,respectively. The ED50 were significant different at 2 Pm and 8 Pm. However, there were no significant difference in ED50 value among other groups. Conclusion The hypnotic effect of ketamine has circadian rhythm - dependent.

OBJECTIVEThe development of neonatology and the availability of pulmonary surfactant have been helpful in effective reduction of the mortality of very low birth weight infants at the expense of an increasing number of survivors with bronchopulmonary dysplasia (BPD) caused by lung immaturity. BPD is a common syndrome in newborns, especially in preterm infants, when treated with hyperoxia and mechanical ventilation. Unfortunately, there have been no effective measure for the prevention and treatment of BPD. The purpose of this study was to investigate the influence of recombinant human insulin-like growth factor-1 (rh-IGF-1) on cell apoptosis and Clara cell secretory protein (CCSP) expression during the lung injury induced by hyperoxia, so as to assess its effect on the inflammatory lung injury and its developmental repair.

METHODSEighty full term neonatal Wistar rats under the same condition were divided randomly into four groups on the second day after birth. Group I was air control, group II was exposed to hyperoxia, group III air + rh-IGF-1, and group IV was treated with hyperoxia + rh-IGF-1. The pups in the control group were kept in room air, while pups in hyperoxia group were kept in a Plexiglas chamber and exposed to over 85% oxygen. Pups in group III were under the same raising condition except for exposure to room air and treated with intraperitoneal injection of rh-IGF-1 (1 microg/Kg) everyday from the third day. Pups in group IV were treated with intraperitoneal injection of rh-IGF-1 (1 microg/Kg) everyday from the third day of exposure to hyperoxia. Lung tissue sections of the neonatal rats were stained with hematoxylin and eosin (HE) after 7 d of hyperoxia exposure, expression of CCSP was examined by immunohistochemical method, and apoptotic cell index of lung tissue was calculated by using TUNEL method.

RESULTSIt was observed from immunohistochemical examination that positive staining of CCSP was distributed mainly in distal and respiratory bronchioles. The percentage of Clara cells in distal and respiratory bronchioles epithelium decreased in hyperoxia group (32.17 +/- 3.19)% compared to that in air control group (68.32 +/- 2.04)%, P < 0.01. Statistically significant differences were found in intensity of positiveness of Clara cells between hyperoxia (29.45 +/- 5.56) and air control group (42.37 +/- 3.24), P < 0.01. TUNEL assay showed that most apoptotic cells were alveolar and bronchial epithelial cells. The apoptotic index increased significantly in the hyperoxia group (55.77 +/- 6.09)% compared to the air control group (16.41 +/- 4.01)%, (P < 0.01). The positive rate (52.98 +/- 2.68)% of Clara cells and the expression (41.22 +/- 6.36) of CCSP in hyperoxia + rh-IGF-1 group increased significantly when compared with hyperoxia group, and the differences between these two group were also statistically significant (P < 0.01). The apoptotic index increased significantly in the hyperoxia + rh-IGF-1 group (27.98 +/- 3.09)% compared to the hyperoxia group (P < 0.01).

CONCLUSIONSHyperoxia exposure can promote the pneumocyte apoptosis and inhibit the expression of CCSP. Rh-IGF-1 can remove the block of the formation of lung alveoli, increase the secretion of CCSP, mitigate inflammatory responses in airway and alleviate lung injury via pneumocyte apoptosis. Therefore, the results of this study provide a theoretic and experimental evidence for clinical application of rh-IGF-1 in prevention and treatment of BPD.

Alveolar Epithelial Cells ; metabolism ; Animals ; Apoptosis ; Epithelial Cells ; Humans ; Hyperoxia ; metabolism ; pathology ; Infant, Newborn ; Insulin-Like Growth Factor I ; genetics ; metabolism ; Lung ; Oxygen ; metabolism ; Rats ; Rats, Sprague-Dawley ; Rats, Wistar ; Uteroglobin ; metabolism

To clone cDNA of human leptin gene and obtain leptin protein for future study on leptin binding proteins. The cDNA of human leptin with 6 x his-tag was cloned by over-hang extension PCR protocol using human genomic DNA as template, and subcloned into in vitro expression vector pIVEX2.3MCS, and the fusion protein was expressed in vitro by Rapid Translation System (RTS) (RTS500 cycle primer Kit and RTS500 ProteoMaster of Roche company). The apparent molecular weight(19.46 kD) and the immuno-specificity of the fusion protein were confirmed by SDS-PAGE and Western blot, and the expressed fusion protein stayed mainly in the supernatant of the reaction mixture in soluble form. This work provides us solid basis for further study on new leptin-associated proteins.
Blotting, Western ; Cloning, Molecular ; DNA, Complementary ; genetics ; Electrophoresis, Polyacrylamide Gel ; Humans ; Leptin ; chemistry ; genetics ; metabolism ; Molecular Weight ; Polymerase Chain Reaction ; Recombinant Fusion Proteins ; chemistry ; genetics ; metabolism

OBJECTIVETo investigate the effect of insulin-like growth factor-1 (IGF-1), which can promote cell differentiation and inhibit cell apoptosis, on hyperoxia-induced apoptosis in A549 cells and its anti-apoptotic mechanism.

METHODSA549 cells were sub-cultured, exposed to hyperoxic conditions and were then treated with different concentrations of IGF-1 (1, 10, and 100 ng/mL) for 48 hours. Cell viability was measured by MTT assay. Cell apoptosis was evaluated by Annexin V-FITC/PI double-staining flow cytometry. Expression levels of Bax and Bcl-2 were measured by flow cytometry.

RESULTSThe middle-dose and high-dose IGF-1 intervention groups had higher cell viabilities than the hyperoxic exposure group [(64±3)% and (88±4)% vs (51±3)%; P<0.05]. Compared with the air control group, the hyperoxic exposure group had a significantly higher apoptotic rate [(38.3±5.4)% vs (2.4±0.9)%; P<0.05], a significantly lower expression level of Bcl-2 [(72±5)% vs (91±4)%; P<0.05], and a significantly higher expression level of Bax [(54±6)% vs (3±2)%; P<0.05]. Compared with the hyperoxic exposure group, the low-dose, middle-dose, and high-dose IGF-1 intervention groups had significantly lower apoptotic rates [(16.1±4.7)%, (9.2±2.8)%, and (6.9±2.5)% vs (38.3±5.4)%; P<0.05], significantly higher expression level of Bcl-2 [(79±4)%, (94±4)%, and (100±5)% vs (72±5)%; P<0.05], and significantly lower expression level of Bax [(26±4)%, （5±2)%, and (4±2)% vs (54±6)%; P<0.05].

CONCLUSIONSHyperoxia significantly inhibits proliferation and promotes apoptosis in A549 cells. IGF-1 may promote cell proliferation and inhibit hyperoxia-induced apoptosis in A549 cells by regulating the expression of Bcl-2 and Bax.

Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Survival ; drug effects ; Dose-Response Relationship, Drug ; Humans ; Hyperoxia ; pathology ; Insulin-Like Growth Factor I ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; analysis ; bcl-2-Associated X Protein ; analysis

Objective To learn the pension willingness and influencing factors of empty-nest elderly in Hangzhou City. Methods A total of 1000 empty-nest elderly in Hangzhou, taking a formal hosusehold registration, living≥1 year, age≥ 65 years old, were selected from two urban districts and three suburbs by multi-stage simple random sampling, and were surveyed with questionnaires. The data was analyzed by logistic regression model to study the influencing factors of pension willingness. Results A total of 992 valid questionnaires were collected and the effective rate was 99.20%. There were 407 (41.03%) , 365 (36.79%) , 208 (20.97%) and 12 (1.21%) people in the 992 empty-nest elderly, choosing their children's support, social pension insurance, personal pension and other old-age methods. Multivariate logistic regression analysis showed that, the empty-nest elderly who worried about pension costs (OR=3.007, 95%CI:1.751-5.155), and the greatest wish was physical health (OR=4.404, 95%CI:1.461-13.276), family harmony (OR=7.724, 95% CI: 2.158-27.646), children work smoothly (OR=4.811, 95%CI: 1.203-19.246) . The lower health score (OR=0.982, 95% CI: 0.965-0.999), choosing their children's support as their pension willingness were relatively high, and the empty-nest elderly who worried about pension costs (OR=2.058, 95%CI: 1.267-3.344), the original occupation for the staff (OR=2.353, 95% CI: 1.091-5.078) , city household registration (OR=0.546, 95% CI:0.349-0.856) . The lower the health score (OR=0.979, 95%CI: 0.966-0.993) choose social pension insurance as their pension willingness would be relatively high pension. Conclusion The influencing factors of the pension willingness of the empty-nest elderly were worried about pension costs, the greatest wish, and health status, urban and rural household registration.

Discharge against medical advice (DAMA) conflicts with the purpose of disease treatment in children. Some research has shown that there are high proportions of extremely preterm infants and infants with asphyxia or congenital malformation in neonates with DAMA. This suggests that the sustainable development of neonatology needs cooperation and co-development with obstetrics, neonatal surgery, and radiology to reduce the rate of DAMA. With reference to the current status of research in both China and other countries, this article reviews the causes for DAMA and the strategies for reducing the rate of DAMA, in order to provide a theoretical basis for effectively reducing the rate of DAMA from the neonatal intensive care unit, improving treatment outcomes of the neonates, and increasing hospitals' comprehensive benefits.
Ethics, Medical ; Health Services Needs and Demand ; Humans ; Infant, Newborn ; Insurance, Health ; Intensive Care Units, Neonatal ; Patient Discharge ; Prenatal Care ; Treatment Refusal

OBJECTIVETo investigate the changes in cisplatin sensitivity of resistant ovarian cancer A2780 cells after inhibition of miR-23a expression and explore the molecular mechanisms.

METHODSThe drug-resistant ovarian cancer A2780 cells were exposed to cisplatin alone or in combination with antagomir-23a. The cell inhibition rates after the treatments were detected using MTT assay, cell cycle changes assessed with flow cytometry; and apoptotic cells observed using Hoechst33258 staining. The changes in glycoprotein P-gp expression in the cells were detected using Western blotting.

RESULTSInhibition of miR-23 a combined with cisplatin treatment significantly increased the cell inhibition rate (P<0.01) and lowered the IC(50) so of cisplatin by 83.76% from 110.18 μmol/L in the control group to 17.89 μmol/L (P<0.01). The combined treatments also caused cell cycle arrestin G0/G1 phase, increased the cell apoptosis rate (P<0.01) and the number of cells stained with Hoechst33258; the cellular expression of P-gp protein was significantly reduced as the cisplatin doses increased (P<0.01).

CONCLUSIONInhibition of miR-23a expression increases the sensitivity of A2780 cells to cisplatin possibly by inhibiting the negative regulation by miR-23a target genes that causes inhibition of P-gp protein expression.

ATP-Binding Cassette, Sub-Family B, Member 1 ; metabolism ; Antineoplastic Agents ; pharmacology ; Apoptosis ; Cell Cycle ; Cell Line, Tumor ; drug effects ; Cisplatin ; pharmacology ; Drug Resistance, Neoplasm ; Female ; Humans ; MicroRNAs ; metabolism ; Ovarian Neoplasms ; pathology