Objective To study the protective effect of paeonol against hypoxic-ischemic brain damage of neonatal rats. Methods 36 neonatal SD rats were randomly divided into normal control group, sham group, HIBD group, and HIBD plus Pae group. The expression of cleaved caspase -3 in the damaged cortex of rats was examined by the immunohistochemical staining and the number of necrotic neurons was counted after hematoxylin-eosin staining. Results The number of the cleaved caspase - 3 positive cells in the damaged cortex of rats in the HIBD plus Pae group was significantly lower than that in the HIBD group( P < 0.05 ). The percentage of necrotic neurons was decreased in the HIBD plus Pae group in comparison with HIBD group( P <0.01). Conclusions Paeonol may take protective effect against hypoxic-ischemic brain damage.

Objective To investigate Treg and Th17 cell numbers in the peripheral blood of patients with unexplained recurrent spontaneous abortion (URSA),with an aim to elevate their potential for predicting pregnancy outcomes.Methods Peripheral blood was collected from 60 URSA pregnant women and 56 healthy pregnant women during their first trimester in the Department of Reproductive Endocrinology of the Second Affiliated Hospital of Soochow University from January 2014 to June 2015.The counts of regulatory T cell (Treg),T helper cell 17 (Th17),CD4 positive T cell (CD4+ T cell) and total lymphocytes in the peripheral blood were determined by flow cytometry.Calculated the ratios of Treg/CD4+ T cells,Treg/total lymphocytes,Th17/CD4+ T cells,Th17/total lymphocytes and Treg/Th17,then the pregnancy outcomes were followed up.According to the different pregnancy outcomes,the patients were divided into three groups: URSA successful delivery group (23 cases),URSA re-miscarriage group (37 cases) and normal successful delivery group (44 cases).Non-parametric test were used to compare the differences between groups.Results There were no difference between the URSA re-miscarriage group and URSA successful delivery group in the CD4+ T cell counts[(1 570.16±519.56) vs (1 571.83±450.37),t=0.01,P=0.99]and total lymphocytes[(4 497.11±975.48) vs (4 558.52±1 032.54),t=0.23,P=0.82].However,the ratios of Treg/CD4+ T cells[(1.09±0.62) vs (3.32±2.30),t=4.55,P=0.000 1],Treg/total lymphocytes[(0.42±0.24) vs (1.31±1.14),t=3.67,P=0.001 3]and Treg/Th17[(0.56±0.41) vs (2.51±2.38),t=3.89,P=0.000 8]in the URSA miscarriage again group were significantly lower whereas the ratios of Th17/CD4+ T cells[(1.85±0.92) vs (0.93±0.36),t=5.40,P<0.001]and Th17/total lymphocytes[(0.64±0.31) vs (0.31±0.14),t=5.52,P<0.001]were significantly higher.Similarly,There were no difference between the URSA re-miscarriage group and normal successful delivery group in the CD4+ T cell counts[(1 570.16±519.56) vs (1 566.86±651.36),t=0.03,P=0.98]and total lymphocytes[(4 497.11±975.48) vs (4 720.91±1 572.92),t=0.78,P=0.44].However,the ratios of Treg/CD4+ T cells[(1.09±0.62) vs (3.17±1.81),t=7.16,P<0.001],Treg/total lymphocytes[(0.42±0.24) vs (1.03±0.51),t=7.08,P<0.001]and Treg/Th17[(0.56±0.41) vs (1.87±1.68),t=4.98,P<0.001]in the URSA re-miscarriage group were significantly lower whereas the ratios of Th17/CD4+ T cells[(1.85±0.92) vs (1.12±0.50),t=4.29,P<0.001]and Th17/total lymphocytes[(0.64±0.31) vs (0.36±0.15),t=5.01,P<0.001]were significantly higher.Conclusions Treg and Th17 cell counts in the peripheral blood of patients with URSA might be useful in predicting their pregnancy outcomes.The decrease of Treg cell counts and the increase of Th17 cell counts might imply another abortion.

In this research, Casuarina eguisetifolia Linn was used to verify the broadly suitability of DNA bar-codes in identification of Li-medicine plants and systematic development of species. The genomic DNA of 22 samples collected C. eguisetifolia and its adulterants were amplified by 4 pairs of primers respectively (ITS (inter-nal transcribed spacer), ITS2 (internal transcribed spacer 2), trnH-psbA , rbcL) and sequenced bi-directionally. Obtained sequences were assembled using CodonCode Aligner. The dates were analysised using MEGA5.1 in ac-cordance with the kimura 2-parameter (K2P) model. The neighbor-joining (NJ) phylogenetic trees were construct-ed. Our study demonstrated the efficacy of ITS/ITS2 to distinguish between C. eguisetifolia and other adulterants species at the molecular level. Comparative to the primer of trnH-psbA and rbcL, there was a obviously DNA gap. The NJ trees showed that the several species of Casuarina can be classified to same types to show a obvi-ously monophyly, which the nearest family was Guttiferae. Therefore, ITS/ITS2 regions can accurately distinguish the original plant of Li-medicine. The systematic evolution of Casuarina can be verified in the molecular level.

Objective To screen cell growth and senescence-related genes of the parasitic pmtist Trichomonas vaginalis,we launched an EST program and isolated two cDNA clones from a T.vaginalis cDNA library,which showed high homology in deduced amino acid sequences to yeast Sir2 and designated as TvSir2 and TvSir2-like.Method The cDNA sequence of TvSIR2 had a length of 1034 base pairs (bp) with an open reading frame of 915 bp,and TvSIR2-like,1214 bp with an open reading frame of 1116 bp.Result The two deduced amino acid sequences shared all the three conserved cole domains with yeast Sir2 and its homologues,suggesting that the two clones were Sir2 homologues. A cDNA fragment from each cDNA clone was subvloned into the expression vector pET-41a.The expression of the fusion proteins in E.coli BL21 stains was induced by isopropylthio-β-D-galactoside (IPTG).Two anti-sera were prepared by immunizing two guinea pigs with the purified fusion proteins, Western-blot analysis demonstrated that each anti-serum reacted with the corresponding recombinant protein and detected a clear band (TvSir2,34 000 Mr;TvSir2-like,42 000 Mr)in protein extracts of the protist.Immunofluolescence techniques showed that TvSir2 and TvSir2-like proteins were both localized in the legions of perinuelear (ER) and Golgi complex.Conclusion Our data suggest that TvSir2 and TvSir2-like were two members of Sir2 family.Their biological functions in the protist would be further studied.

Objective To study the effects of hypoxia on the expression of inflammatory factor high mobility group box-l(HMGB1) in the pulmonary arteriolae of neonatal SD rats.Method A total of 80 neonatal SD rats were randomly assigned into control group and hypoxia-induced persistent pulmonary hypertension of the newborn model (PPHN) group.The PPHN group was subdivided into 2 h,8 h,24 h,and 3 d post-PPHN subgroups according to the time of sacrifice.PPHN model was established on postnatal day 4 when rat pups in PPHN group were kept in low-oxygen box (10％ O2 and 90％ N2) for consecutively 7 days.Multi-channel physiological transducer RM-6280 was used recording the mean pulmonary artery pressure (mPAP) at the root to pulmonary artery of rat pups.ELISA method was used examining the serum level of HMGB1 of rat pups in each group.The pathology of the lung tissue was studied using optical microscope after HE staining,and MIAS-2000 medical image analysis software was used to calculate the ratio of the middle membrane thickness to the outer diameter of the pulmonary arteriolae wall (MT％).Protein level of HMGB1 in the lung was examined using Western Blot.Result The lung pathology in PPHN rats showed thickening of the middle membrane of the pulmonary arteriolae wall and stenosis of the pulmonary arteriolae.MT％ of control group and PPHN group were 5.3％ (3.7％,7.6％) and 7.1％ (4.6％,9.2％),respectively,without significant differences (P＞0.05).At 2 h,8 h,24 h,3 d post-PPHN timepoints,the serum levels of HMGB1 in PPHN group were (13.2±3.1),(15.4±3.6),(17.1±3.5),and (15.8±3.6) ng/ml,respectively,without intra-subgroup differences (F=2.134,P＞0.05),but significant differences existed when compared with control group at each timepoint (P＜0.01).Western Blot showed that HMGB1 protein expression in the lungs were significantly elevated soon after PPHN,peaked at 8～24 h,and reduced but still significantly elevated at 3 d after PPHN comparing with normal control.Significant differences existed at 2 h,8 h,and 24 h timepoints (P＜0.01,respectively).The HMGB1 protein of PPHN group declined significantly at 3 d timepoint without significant differences comparing with the control group (P＞0.05).Conclusion HMGB1 is closely related with the pathogenesis of PPHN,indicating the inflammatory response plays an important role in the mechanisms of PPHN.HMGB1 may be an indicator for the assessment of hypoxia-induced PPHN.

Objective To study the characteristics of amplitude integrated electroencephalogram (aEEG) in full-term newborns with different blood glucose levels,so as to provide clinical evidence for assessing brain function after hypoglycemia.Method Full-term neonates admitted to the neonatal ward of the Third Xiangya Hospital of Central South University from June 2014 to May 2016 with the initial diagnosis of hypoglycemia were enrolled to hypoglycemia group.According to the lowest level of blood glucose,infants were assigned to three subgroups,severe hypoglycemia group (＜ 1.1 mmol/L),moderate hypoglycemia group (1.1 ～ ＜2.2 mmol/L),and mild hypoglycemia group (2.2 ～ ＜2.8 mmol/L).Time matched asymptomatic term infants,who were admitted to the neonatal ward due to maternal high risks and with normal blood glucose after birth,were enrolled to control group.A 4 h continuous aEEG monitoring was completed for each infant in hypoglycemia group within 12 h after the blood glucose level stabilized.The newborns in control group were given aEEG examination 72 ～ 120 h after birth,the duration of monitoring was also 4 h.The aEEG scoring was completed and compared by rank sum test.Result A total of 83 neonates were enrolled in hypoglycemia group,including 11 with severe hypoglycemia,32 with moderate hypoglycemia,and 40 with mild hypoglycemia.Another 26 neonates with normal blood glucose level were enrolled in control group.The incidence of pregnancy-induced maternal blood glucose elevation was statistically significant among each group (P ＜ 0.05).The duration of neonatal hypoglycemia in severe hypoglycemia group was longer than that in moderate hypoglycemia group and mild hypoglycemia group [38.3 (20.7,50.4) h vs.20.4(15.3,22.6) h,13.7 (7.8,19.4) h] (P＜ 0.05).The range of glucose level in severe hypoglycemia group was larger than that in mild and moderate hypoglycemia group [5.0 (4.0,5.5) mmol/Lvs.3.5 (3.0,3.9) mmol/L,3.3 (2.8,3.8) mmol/L] (P ＜ 0.05),but there was no significant difference in the onset of first hypoglycemia between groups (P ＞ 0.05).The aEEG score showed that there was significant difference in total score and sleep-wake cycle score between groups (P ＜ 0.05).The score of sleep-wake cycle in severe hypoglycemia group was significantly lower than that in moderate hypoglycemia group or in mild hypoglycemia group or in the control group (P ＜ 0.05),while there was no significant difference between moderate and mild hypoglycemia groups,and between moderate hypoglycemia and control group (P ＞ 0.05).Conclusion Severe hypoglycemia can lead to neonatal aEEG changes,mainly in the sleep-wake cycle changes.