Objective:To investigate the mechanism of inducing immune tolerance of autoimmune diabetic mice by insulin B chain gene vaccine Methods:Insulin B chain DNA vaccine was constructed by genetic engineering techniques DNA vaccine was injected into tibilias anterior muscle of C57BL/6 mice Mice were divided into four groups at random: group control (C), group diabetes (D), group treatment (T), prophylactic treatment (P) Serum was taken to determine insulin, the expression of bcl 2 mRNA was determined by dot blot dehydribation Results:①After prediabetic mice were injected by DNA vaccines, the ratio of incidence of diabetes was 30% in 2 w, 40% in 3 w and 4 w, which were significantly higher than that of groups D and T, and lower than that of group C The level of insulin showed no difference between group C and the non diabetic mice of group P In contrast to non diabetic mice, the change of pancreas of diabetic mice in group P was similar to group D ②AGV of bcl 2 mRNA in thymus cells, spleen cells, and blood monocytes of effective mice in group P showed no difference with group C, but markly lower than those of group D & T Conclusion:Insulin B chain DNA vaccine could recover the balance of the development and activity of thymus cells, spleen cells and lymphocyte in prediabetic mice

Objective To explore the dynamic changes of D-dimers during pregnancy and early puerperium (within 3 days postpartum). Methods A retrospective study was performed among 8 367 healthy women who had term singleton delivery in Women′s Hospital, School of Medicine, Zhejiang University from January 2007 to December 2014. D-dimers concentrations during pregnancy and early puerprium of all the cases were collected. Data of 21 065 D-dimers tests were assigned to 5 groups according to the time of sampling, including early pregnancy (≤12 gestation weeks), middle pregnancy (12-28 gestation weeks), late pregnancy (>28 gestation weeks), 1 postpartum (within 48 hours postpartum) and 2 postpartum (48-72 hours postpartum). The D-dimers concentrations in different groups were compared. The effect of delivery mode on D-dimers of early pureperium was analyzed. The correlation between D-dimers and the thromboembolic disease was also explored. In this study, Student′s t-test and Wilcoxon rank sum test were used for statistical analysis. D-dimers concentration≤0.5 mg/L was used as the normal range. Results (1) D-dimers concentrations during pregnancy were higher than the non-pregnant women (P<0.01), but there was no statistical difference between early pregnancy and late pregnancy (P=0.820). D-dimers concentration in the 1 postpartum group was higher than that of early pregnancy group or late pregnancy group (P<0.01). But in the 2 postpartum group, it was lower than early pregnancy, late pregnancy and 1 postpartum groups. (2)D-dimers in cesarean section cases was significantly higher than in vaginal delivery cases in each period of pregnancy and early pueprium.(3)The 95%CI of D-dimers in early pregnancy, late pregnancy, 48 hours after vaginal delivery, 48-72 hours after vaginal delivery, ≤48 hours after cesarean section, 48-72 hours after cesarean section were 0.58-8.28, 0.47-11.52, 1.04-9.59, 0.87-5.22, 1.07-11.58 and 1.00-6.23 mg/L, respectively.(4)In 6 cases with thromboembolic disease, D-dimers was 6.89-19.89 mg/L, with the mean value of 13.66 mg/L. It was significantly higher than normal range. In 3 cases, all after cesarean section, with lower extremity vein thrombosis within 48 hours postpartum, the D-dimers concentrations, 9.77, 8.65 and 6.89 mg/L respectively, were in the 95%CI of the study population after cesarean section. Conclusions D-dimers concentration of 0.5 mg/L is not suitable for venous thromboembolism screening during pregnancy. D-dimers concentration in pregnancy and early puerprium is higher than non-pregnancy. It increases in the very early period postpartum and decreases with time. D-dimers should not be a routine screening test to exclude thromboembolic disease in pregnant women without high risk factors and clinical manifestation of thromboembolic disease.

Objective To investigate the relationship between the expression of glucose regulated protein 78 (GRP78) in the placental trophoblast cells and the pathogenesis of gestational diabetes mellitus (GDM).Methods All the patients were recruited from Qingdao Municipal Hospital from May 2013 to May 2014.Among them, fifty women with GDM were assigned to the GDM group, and fifty healthy women were defined as the control group.All of them received cesarean section because of breech presentation, contracted pelvis, scarred uterus or on mother's demand.Real-time PCR was conducted to analyze the expression of GRP78 mRNA in the trophoblasts.Immunohistochemistry was performed to detect the localization of GRP78 protein in the placentasl trophoblast cells.Results (1) GRP78 mRNA expressed in the cytoplasm of trophoblasts of both the GDM group and the control group.The GRP78 mRNA levels in the GDM group and the control group were 15.6±0.4 and 6.0±0.7, respectively.The relative expression level of GRP78 mRNA in the GDM group was 2.6 times of that in the control group, with statistically significant difference (P＜0.01).(2) The expression of GRP78 protein was found in the cytoplasm of the trophoblasts of the GDM group.It showed in deep, light brown or yellow after staining, according to the expression degree.The expression of GRP78 protein was also found in the cytoplasm of the trophoblasts of the control group, but it mainly showed yellow color (38/50).The strong positive rate of GRP78 protein in the GDM group (96％, 48/50) was higher than that in the control group (22％, 11/50;P＜0.01).Conclusion The expression of GRP78 increased in the placental trophoblast cells of GDM patients.It might suggest that GRP78 had some effect on the pathogenesis of GDM.

Objective To evaluate the predictive accuracy of several risk-assessment strategies to predict the risk of significant neonatal hyperbilirubinemia, and to establish the best prediction model.Methods The transcutancous bilirubin (TcB) levels of 4907 term and near-team infants were measured.Trace blood bilirubin levels of the infants whose TcB levels ≥250 μmol/L were detected. Clinical data of newborns and their mothers were collected and were analyzed with Logistic regression model to investigate its correlation with signifrcant hyperbilirubinemia. Clinical high risk factors of significant neonatal hyperbilirubinemia were determined. Accuracy of three prediction methods for significant hyperbilirubinemia was compared by receiver operating characteristic (ROC) curve. The three methods included: whether predischarge bilirubin level (within 72 hours after birth) expressed in risk zone on an hour-specific bilirubin nomogram; clinical risk factors other than predischarge bilirubin level; and combination of the predischarge bilirubin risk zone and other clinical risk factors. Results Two hundred and eighty-six newborns (5.8%) were found with significant hyperbilirubinemia. The risk factors of significant neonatal hyperbilirubinemia were divided into three groups according to OR: (1) Major risk factors:predischarge (within 72 hours after birth) bilirubin level in the high risk-zone (OR=96. 39, 95% CI:53.32-174.27, P = 0. 000), large cephalohematoma (OR = 36.45, 95% CI: 10. 02-132.56,P=0. 0076), gestational age 35-36+6 weeks (OR= 30. 72, 95% CI 14.47-65.23, P=0. 0001) and exclusive breast feeding and weight loss was ＞9% of birth-weight (OR=22.44, 95% CI: 4.42-114. 03, P=0. 0016). (2) Minor risk factors: gestational age 37-37+6 weeks (OR=3.26, 95% CI:1.92-5. 55, P=0. 0232), predischarge bilirubin level in P76-P95(OR=13. 64, 95% CI: 8. 10-22.97,P=0. 0001) and bruising (OR = 2.32, 95% CI: 1.14-4.71, P = 0. 0497). (3)Protective factors (those factors associated with decreased risk of hyperbilirubinemia): predischarge bilirubin level in low-risk zone (≤P40) (OR=0. 00), gestational age ≥40 weeks (OR=0.21, 95% CI: 0.09-0.44,P=0. 0402) and mixed breeding (OR=0. 75, 95% CI: 0. 58-0.95, P=0.0059). The area under the ROC curve of predischarge bilirubin level was 0. 8687 and 0. 7375 for clinical risk factors other than predischarge bilirubin level. The area under the ROC curve of a combination of the predischarge bilirubin risk zone and additional clinical risk factors was 0. 9367. Conclusions The risk of significant neonatal hyperbilirubinemia could be simply and accurately predicted by infant's predischarge bilirubin level and the combination of predischarge bilirubin level, and clinical risk factors might improve the accuracy of prediction significantly.

Objective To evaluate the role of M3 muscarinic acetylcholine receptor (M3 mAChR) in the release of glycinergic neurotransmitter by using oxotremorine-M (Oxo-M: a nonselective mAChR agonist) and 4-diphenylacetoxy-N-methylpiperidine methiodide (4-DAMP: a highly selective M3mAChR antagonist). Methods Twenty male 3-4 weeks old SD rats weighing 160-180 g after successful intrathecal catheterization were randomized into 2 groups (n = 10 each): normal saline group (group NS) and pertussis toxin (group PTX).Pertussis toxin 1.5 μg/10 μl was injected IT in group PTX, while in group NS normal saline 10 μl was injected IT instead. The animals were killed at day 7 after injection. The spinal cords were removed and sliced and placed in artificial CSF. Glycinergic spontaneous inhibitory postsynaptic currents (sIPSCs) were measured in spinal lamina Ⅱneurons using whole-cell voltage-clamp technique. Five minutes after sealing, Oxo-M (final concentration 3 μ mol/L) was added. Oxo-M was then completely washed out 3 min later and 4-DAMP (final concentration 25 nmol/L) was added after 5 min of stabilization. In the presence of 4-DAMP, Oxo-M (final concentration 3 μmol/L) was added again 3 min later. sIPSCs were recorded before addition of Oxo-M (T1), 3 min after addition of Oxo-M (T2), 3 min after addition of 4-DAMP (T3), 3 min after the second addition of Oxo-M (T4). Results Compared with the baseline value at T1 , Oxo-M significantly increased the frequency of glycinergic sIPSCs at T2without changing the amplitude at T2-4 in both groups. The frequency of sIPSCs was significantly lower at T4 than at T2 in both groups (P ＜ 0.05 or 0.01). There was no significant difference in both frequency and amplitude of glycinergic sIPSCs between the two groups. Conclusion M3 mAChR plays a predominant role in the release of glycinergic transmitter in the spinal lamina Ⅱ neurons in rats.

Objective To study the expression and significance of multidrug resistance gene (MDR1) in children with refractory epilepsy (RE). Methods Children with RE (n = 30), non-RE (n = 30) and healthy children (n=30) were collected. The expression of MDR1-mRNA in peripheral blood was analyzed by fluorescence quantitative PCR. The relationship of MDRI-mRNA with epileptic frequency and numbers of antiepileptic drugs (AEDs) were observed. Results The expression of MDR1 in RE group obviously increased when compared with that of non-RE group and healthy group (P < 0.01, P < 0.01) ; MDR1 expression was more among patients with high frequent epilepsy than patients with low frequent epilepsy (P < 0.01) ; more in patients administered with four kinds of AEDs than those with two or three kinds of AEDs (P < 0.01). Concinsions MDR1 overexpression in blood of children with RE may be linked to drug-resistant mechanism of RE. It might be used as a clinical indicator of RE.

Objective To systematically evaluate the effectiveness of delayed cord clamping (DCC) in term infants. Methods The data of the Cochrane library, PubMed, EMBASE, CNKI , VIP, Wanfang from 1 January 1970 to 30 April 2013 were searched. Randomized controlled trials (RCT) of DCC in term infants were included.RevMan 5.1.0 was used in the statis-tical analysis. Results Ten studies involving 1623 participants were included. Meta-analysis based on included studies showed that:compared with immediate cord clamping (ICC), DCC improved the hemoglobin levels at birth (MD=2.19, 95%CI:0.36, 4.02) and increased the incidence of polycythaemia (RR=2.87, 95%CI:1.24, 6.62). Compared with ICC, DCC showed no signi-ficant difference in the phototherapy for hyperbilirubinemia (RR=2.46, 95%CI: 0.93, 6.52), the hemoglobin levels within 6 months (MD=0.29, 95%CI:-0.17, 0.75), and the incidence of anemia (RR=0.71, 95%CI:0.45, 1.12). Conclusions DCC can improve the hemoglobin level in term infants after birth. However, the appropriate time of cord clamping has not been deter-mined. It is necessary to undertake further studies with higher quality and larger scale to evaluate the optimal time of cord clam-ping.

Objective To observe the effect of medicinal wine regimen which is guided by the theory of constitution distinction of TCM.Methods 76 patients were selected from all patients enrolled in wine style health experience from February 2011 to December 2011 in line with the corresponding physical characteristics.Among these 76 patients,23 patients were Yang deficiency constitution,34 were phlegm dampness constitution,and 19 were yin deficiency constitution.All these patients were taken a physical and chemical examination after taking the medicinal wine for a month.Results ①Examination index comparison before and after the treatment in different physical constitution patients:in phlegm dampness constitution patients the pulse were slowed (t=2.122,P＜0.05) after taking the blood fat reducing wine,and physical type of conversion points were lowered (t=4.86,P＜0.01); in Yang constitution patients,the systolic blood pressure (FT4),free thyroxine T4,physical type of conversion points were all significantly decreased [t values were 2.735,3.672,12.93,P＜0.05 or 0.01] after taking Guilu Erxian wine; in Yin deficiency patients,adrenocorticotropic hormone (ACTH),interleukin-1 (IL-1),interleukin-2 (IL-2) were all increased [t values were 2.355,2.645,3.829,P＜0.05 or 0.01] after taking Ziyin-Buxue wine.② Comparison of the clinical curative effect:according to the type of constitution transformation changes,the total effective rate was more than 70％.According to Ridit test,except yin deficiency physique (t=1.85,P＞0.05),patients in both phlegm dampness constitution and yang deficiency constitution groups showed statistical difference after the treatment than before (t=4.86 and 12.93,P ＜0.01).Conclusions Blood fat reducing wine can improve phlegm dampness constitution; Ziyin-Buxue wine can improve Yin deficiency constitution and enhance immune function; Guilu Erxian wine can lower blood pressure and improve Yang deficiency constitution.

The total effective rate for clinical symptomstreated with oral decoction with Weiweikang(benefit-ting atrophic stomach)granules and Huoli(vitality)Bolus guided by the principle of invigorating Qi,warming the interior,activating circulation and elimi-nating blood stasis,was 91.8%,while that for thechronic atrophic gastritis with metaplasia and atypicalhyperplasia of intestinal epithelium was 87.5% and74.4% respectively.After treatment,the volume ofblood flowing was markedly increased(P

Objective To study the efficacy of T-piece resuscitator on the very preterm infants in the delivery room.Method Very preterm infants (gestational age 28 ～ 31 weeks) who needed positive pressure ventilation during delivery room resuscitation were included in the study between January 2010 and December 2015.Enrolled infants were randomly assigned to self-inflating bag group and T-piece group.Tracheal intubation ratio,duration of mechanical ventilation,continuous positive airway pressure (CPAP),supplementary oxygen through a nasal cannula and total oxygen requirement were compared between groups.The percentages of pneumothorax,sepsis,necrotizing enterocolitis (NEC),bronchopulmonary dysplasia (BPD),retinopathy of prematurity (ROP),intracranial hemorrhage and patent ductus arteriosus (PDA) between groups were also compared.Data were analyzed using independent sample t test and chi-square test.Result A total of 51 preterm infants were enrolled in this study,with 25 infants in the self-inflating bag group and 26 in the T-piece group.There was no statistically significant difference in the gender,gestational age,birth weight,Apgar scores,delivery mode and antenatal glucocorticoids between the two groups (P ＞ 0.05).The ratio of intubation in T-piece group was significantly lower than that in self-inflating bag group (15.4％ vs.44.0％,P ＜ 0.05).Further more,duration of mechanical ventilation and total oxygen requirement in the T-piece group were significantly shorter than those in the self-inflating bag group [(4.2±2.8) dvs.(10.1 ±4.3) d,(36.2±14.7) dvs.(47.2±19.2) d,P＜0.05].However,the duration of nasal CPAP and supplementary oxygen through a nasal cannula,the rate of pneumothorax,sepsis,NEC,BPD,ROP,intracranial hemorrhage and PDA did not differ significantly between groups (P ＞ 0.05).Conclusion Compared with the self-inflating bag group,the use of the T-piece in delivery room decrease the rate of tracheal intubation and the duration of mechanical ventilation and total oxygen requirement.