The changes of retinal nuclear DNA content in rats after death was detected and the relationship between degradation of retinal nuclear DNA and postmortem interval (PMI) was analyzed. Ninety healthy adult SD rats, female, weighing 250+/-10 g, were randomly divided into 15 groups. At 20 degrees C, the retinal cells were withdrawn every 2 h within 0 to 28 h after death and stained with Feulgen-Vans. Index of density (ID), integral absorbance (IA) and average absorbance (AA) in retinal nucleus were analyzed by image analysis system. And the obtained data were subjected to linear regression analysis by using SPSS12.0 software. The results showed that in retinal nucleus, AA and IA were gradually declined with the prolongation of PMI, while ID had an increased tendency. Within 28 h after PMI, the regression equations were as follows: Y(AA )=-0.009X(AA )+0.590 (R(2)=0.949), Y(IA )=-0.097X(IA )+18.903 (R(2)=0.968), Y(ID)=0.122X(ID)+2.246 (R(2)=0.951). It was concluded that retinal nuclear DNA after death in rats was degraded gradually and had a good correlation with PMI.

Objective To explore the differences in maternal and neonatal complications of twin pregnancies in different modes of conception.Methods Retrospective analysis was performed to compare the differences in mother's gestation history,childbirth and neonatal complications among 102 cases of in vitro fertilization-embryo transfer(IVFET) pregnancy twins,16 cases of simple ovulation induction(OI) pregnancy twins and 96 cases of spontaneous conception(SC) pregnancy twins.Results 1.The maternal age[(32.51 ±4.03) years vs (31.25 ±3.38) years vs (30.20 ± 4.13) years,F =8.061,P ＜ 0.05] and birthrate of full-term infants (21.5 ％ vs 0 vs 10.4％,x2 =7.566,P ＜ 0.05) of IVF-ET group were significantly higher than that of OI group and SC group.The body weight at birth of OI group was significantly lower than that of IVF-ET group and SC group [(1 918.1 ±516.4) g vs (2 228.3 ±4542) g vs (2 164.4 ±442.5) g,F =3.293,P ＜0.05].The birthrate of late preterm infant(87.5％ vs 51.0％ vs 64.6％,x2 =9.322,P ＜ 0.05) and the incidence of twin discordance (50.0％ vs 21.6％ vs 18.8％,x2 =7.781,P ＜ 0.05) of OI group were significantly higher than that of IVF-ET group and SC group,while the incidence of monozygotic twins of SC group was significantly higher than that of IVF-ET group and OI group(52.1％ vs 5.9％ vs 0,x2 =58.016,P ＜ 0.05).2.The incidence of pregnancy induced hypertension of OI group was significantly higher than those of IVF-ET group and SC group(50.0％ vs 17.6％ vs 22.9％,x2 =8.344,P ＜ 0.05).The incidence of gestational diabetes mellitus of OI group and spontaneous group was significantly higher than that of IVF-ET group(12.5％ vs 14.6％ vs 3.9％,x2 =6.838,P ＜0.05).3.The incidence of twin transfusion syndrome of SC group was significantly higher than that of the other groups(6.3％ vs 0 vs 0,x2 =7.588,P ＜0.05),but the incidence of apnea of prematurity of OI group was significantly higher than that of IVF-ET group and SC group(43.8％ vs 15.7％ vs 24.0％,x2 =7.081,P ＜0.05).The incidence of asphyxia,transient tachypnea of the newborn,hypoglycemia,intracranial hemorrhage and septicemia of OI group were higher than that of IVF-ET group and SC group,but there was no significantly difference among 3 groups.Conclusions IVF-ET conceived twin pregnancies are not at greater risk for pregnancy complications and adverse perinatal outcome than SC twin pregnancies.But the incidence of some adverse maternal and neonatal complications of simple OI twin pregnancies is higher than that of IVF-ET group and SC group.More attention should be paid to the perinatal health care of OI conception twins,which is important to avoid serious complications.

Objective To study the effects of cultivation methods on growth dynamics in the aerial part and root of Angelica sinensis and Ditylenchus destructor, rate of early bolting, and yield of A. sinensis as well. Methods Compared with conventional planting, ridge planting, furrow planting, white plastic mulch coverage, and black plastic mulch coverage were used to evaluate the effects of cultivation methods on dry matter accumulating and growth dynamics of A. sinensis. Results The influence of black plastic mulch coverage to growth dynamics of A. sinensis is the biggest, next for white plastic mulch coverage. There is the best inhibition to D. destructor under ridge planting. Black plastic mulch coverage can enhance the rate of early bolting. The treatment of black plastic mulch coverage has the highest yield, which can increase the yield by 33.4% (4 650 kg/hm2) compared to conventional planting; next for the treatment of white plastic mulch coverage, which increased the yield by 24.3% (3 372.2 kg/hm2) compared to conventional planting. But there were no significant effects of ridge planting and furrow planting on increasing yield. Conclusion The cultivation methods of black plastic mulch coverage should be brought into wide use in A. sinensis cultivation.

Forensic medical talents with creative ability should have ability to find.analyze and solve problems from complex cases.According to teaching practice on the basis of PBL this article illustrates application methods of PBL teaching characters in the training of forensic medical talents with creative ability.

Objective:To analysis the Mimotopes of the peptide mimics to PGN using online softwares.Methods: Mimotopes of PGN were screened from 12-mers linear phage display peptide library by using anti-PGN McAb and the antigenicity of selected clones was identified by ELISA.The B cell epitopes,T cell epitopes of ′GRWxHxVxWAGL′ were estimated by DNAstar and online softwares.Results: 16 phage clones that bound with anti-PGN McAb were screened from 12-mers linear phage display peptide library.Among these positive clones,phage clones No.39 shared the conserved sequence:WxHx……AGL found in previous clone No.31(ATWxHxLxSAGL),which provoked an effective protective immunity against infection with S.aureus.To enhance the stability of the conformation as well as adding biotin on the N-ter-minal as a tag,the sequences ′39′ were redesigned and synthesized by adding S(serine)A(alanine) and GG(glycine)on the C-terminal of origin sequence(named SP39).Next,we estimated or predicted antigenic epitopes,T cell epitopes and scores binding to MHC of these peptides by using DNASTAR and online softwares(http://bio.dfci.harvard.edu/Tools/antigenic.pl,www.syfpeithi.de,http://www.darrenflower.info/mhcpred),indicating that SP39 contains sites bound both mice and human MHC.The sequences ′WxHxVxW-′ may be antigenic epitope as SP39,which contains a T cell epitope.Our results showed that both SP39 could bind to both anti-PGN McAb and a polyclonal antibody against S.aureus.Moreover PGN could inhibit the binding of SP39 to the anti-PGN McAb.These data indicated that SP39 mimic to epitopes on PGN.Conclusion: SP39(GRWxHxVxWAGLAGGS) probably display the mimotopes of PGN.

Objective To observe and evaluate the location of the three compartments of the pelvic floor organ and levator hiatus area in the postpartum women by transperineal real time three-dimensional ultrasound.Methods Sixty eases of maternal with the matching age and body mass index (BMI;postpartum group) and 35 non-fertile women (control group)were selected.The distance between the lower edge and the reference line was measured by the transperineal ultrasound at rest and Valsalva movement in the lowest point of bladder,cervix and rectum.Meanwhile,the area of levator hiatus under Valsalva action was measured by real time 3-dimensional ultrasound.The differences were compared between two groups.Results In the postpartum group,the lowest point of the bladder and cervix was lower than the control group at rest and Valsalva movemen (both P＜0.05).Rectum location in the rest state difference between the two groups had no statistical significance (t=0.405,P =0.398).The area of the levator hiatus of the postpartum group was significantly larger than that of the control group at the rest and Valsalva movement (t=11.253,9.625,P＜0.001).Conclusion Transperineal real time 3-dimensional ultrasound can dynamically observe the position,movement of pelvic floor organ and the area of levator hiatusin of the female.To evaluate the pregnancy and childbirth have obvious effect on the pelvic floor support tissues.

0.05). The rFA2 values were higher and rFA3 were lower in high grade gliomas than that in low grade and had significant difference (respectively t=2.453, P

Objective To investigate the effect of wogonin on ethology and its possible mechanisms in chronic cerebral ischemia in rats.Methods Rats were randomly divided into a sham operation group,a wogonin intervention group,and a phosphate buffered solution (PBS) control group.A rat model of chronic cerebral ischemia was induced by the two-vessel occlusion method.Six weeks after modeling,the rats in the wogonin intervention group and the PBS control group were intragastric administrated with wogonin (50 μmol/L,10 ml/kg,once a day) and PBS with equal volume for 14 days.Morris water maze test was used to evaluate the spatial learning and memory function.Laser confocal three-dimensional vascular imaging was used to detect the vascular proliferation of ischemic brain tissue.5-Bromo-2-deoxyuridine (BrdU)immunochemical staining was used to detect the cell proliferation in ischemic brain tissue.Transmission electron microscope was used to observe the morphological changes of neural cells in cerebral ischernic region.Results The Morris water maze (n =8) showed that the trains of escape latency from the second to the fifth day in the wogonin intervention group were 43.45 ± 8.64 s,37.12 ± 1.31 s,34.75 ± 5.36 s,and 24.36 ± 5.43 s,respectively.They were significantly shorter than 51.69 ± 5.32 s,43.65 ± 9.21 s,50.19 ± 10.31 s,and 53.65 ± 7.15 s in the PBS control group (all P ＜ 0.05).The first quadrant swimming time of the wogonin intervention group was significantly longer than that of the PBS control group (26.16 ±3.29 s vs.14.38 ±2.16 s; P＜0.01).Laser confocal three-dimensional vascular imaging (n=4) showed that the capillary inner diameter in cerebral ischemia region of the wogonin intervention group was reduced significantly compared to the PBS control group (3.02 ±0.21 μm vs.3.35 ±0.18 μm; P ＜0.05),vascular density was increased significantly (205.80 ± 12.70/0.002 mm3vs.158.42 ± 10.92/0.002 mm3; P＜0.01),and total microvascular area was increased significantly (83 389 ± 4 026 μm2/0.002 mm3 vs.73 349 ±3 986 μm2/0.002 mm3; P＜0.01).Immunohistochemical staining (n =6) showed that the number of BrdU positive cells in the ischemic brain tissue of the wogonin intervention group was increased significantly compared to the PBS control group (24.62 ±3.25/HPF vs.9.87 ±2.89/HPF; P＜0.01).The observation of transmission electron microscope showed that the inflammatory edema in the intercellular spaces of the wogonin intervention group was significantly reduced compare to the PBS control group.Conclusions Wogonin can significantly improve the spatial learning and memory ability of chronic cerebral ischemia in rats,and its possible mechanisms may include the promotion of proliferation and angiogenesis in ischemic region and angiogenesis,and reduce inflammatory response.

Objective To explore the clinical significance of a series of cytokines and peripheral blood immunocyte subsets before and after immunosuppressive therapy in patients with immune thrombocytopenia (ITP).Methods The percentages of immunocyte subsets in the peripheral blood of 20 patients with ITP and 20 healthy controls were detected by flow cytometry,including CD3+,CD4+,CD8+,CD4+/CD8+,CD1~.ELISA was applied to detect the level of serum TNFα,IL-2,IL-6,IL-4,IL-10,IL-11,IL-17,IL-27,transforming growth factor β (TGFβ),thrombopoietin (TPO) of 20 patients with ITP and 20 healthy controls.Results The percentage of CD3+ T lymphocyte,CD4+ T lymphocyte and the ratio of CD4+ / CD8+ T lymphocyte in patients with ITP were lower than those in healthy controls [(62.66 ± 6.58) ％ vs (69.93 ± 4.81) ％,(29.46 ± 5.02) ％ vs (39.08 ± 3.50) ％,0.97 ± 0.35 vs 1.56 ± 0.26,all P ＜ 0.05].After immunosuppressive therapy,the percentage of CD3+ T lymphocyte,CD4+ T lymphocyte and the ratio of CD4+/CD8+ T lymphocyte [(71.49 ±5.16)％,(39.25 ±3.21)％ and 1.56 ±0.28] recovered to the same levels in healthy controls.The percentage of CD8+ T lymphocyte and CD19+ B lymphocyte in patients with ITP were higher than those in the healthy controls [(30.28 ±4.63)％ vs (25.90±3.06)％,(18.92 ± 4.27)％ vs (13.17 ± 3.64)％,all P ＜ 0.05].After treatment of immunosuppressive therapy,the percentage of CD8+ T lymphocyte and CD19+ B lymphocyte [(25.16 ± 3.45) ％ and (11.98 ± 3.68) ％] recovered to the similar levels in healthy controls.The serum levels of IL-4,IL-6,IL-11,IL-17 and TPO in patients with ITP were significantly higher than those in healthy controls.While TGFβ level was significantly decreased.There was no significant difference of IL-27 between ITP patients and healthy controls.After the treatment of immunosuppressive therapy,IL-4,IL-6,IL-11,IL-17,TPO and TGFβ were down-regulated while IL-27 was up-regulated.There was no significant difference of IFNγ,TNFα,IL-2 and IL-10 among ITP patients before or after immunosuppressive therapy and healthy controls.Conclusions The present study suggests that the aberrant immunocyte subsets and cytokines are involved in the pathogenesis of ITP.Hyper-function of Th2 and Th17,dysfunction of Treg cells,up-regulation of IL-27,IL-11,TPO and other factors may contribute to the pathogenesis of ITP.

We obtained recombinant ferritin from Dermatophagoides f arinae ,and analyzed the characterization of the pro‐tein .A pair of primers was designed according to the known sequence of ferritin gene .The live mites identified and cultured lo‐cally were picked and the total RNA was extracted .The ferritin gene fragment was amplified by RT‐PCR ,and cloned into pET32a vector ,and then transferred into E .coli Top10 .The target gene obtained from the recombinant plasmid by digestion with Bam HⅠand Hind Ⅲ was connected to the prokaryotic expression vector pET‐32a .The expressed recombinant plasmid containing ferritin gene was constructed by cloning target gene into pET‐32a and transferred into E .coli Bl21 (DE3) .The ex‐pressed recombinant protein was analyzed by SDS‐PAGE ,and was purified by immobilized metal ion affinity chromatography (IMAC) .The ferritin expressed by dust mite was analyzed by the method of bioinformatics .The recombinant plasmid pET32a‐ferritin was constructed .SDS‐PAGE showed a correct molecular weight of the recombinant ferritin protein .After purification by affinity chromatography ,the protein showed only one strip on SDS‐PAGE gel .SDS‐PAGE showed a band at 20 kD .Dust mite ferritin contains 8 serine kinase ,7 threonine kinase ,7 tyrosine kinase ,and 0 histidine kinase phosphorylation sites .Hy‐drophilic region is larger than the hydrophobic region and it is an unstable protein .In conclusion ,the ferritin gene has been cloned and expressed .The purified ferritin has high purity . The study provides a basis for further study of composition and physicochemical properties of house dust mite allergen .