Objective To establish a purified model of rat retinal ganglion cells (RGCs) cultured by serum-free medium,and provide a good cell model to investigate the damage of RGCs in glaucoma,retinal ischemia,and degenerative retinopathy. Methods Two monoclonal antibodies,anti-rat SIRP (OX-41) against rat macrophage and antibody against rat Thy-1 (OX-7),were used to purify and characterize RGCs from 1-3-day old Sprague-Dawley (SD) rats by means of two-step filtration.Purified RGCs were cultured in serum-free neurobasal medium containing B27 and ciliary neurotrophic factor (CNTF) meeting the neuronal cell's special requirements.Photomicrographs illustration,immunfluorescence staining of Thy-1,calcein-acetoxymethyl ester (calcein-AM) fluorescence images were used to observe and identify cultured retinal cells and purified RGCs. Results Among the primary cultured rat retinal cells,91% were retinal neurons.Protuberances of RGCs were seen after cultured for 24 hours.At the4th to 8th day,many cells had uniform configuration,large body,and long protuberances. At the 14th day,over 60% cells maintained viability.Immunoflurescence staining of Thy-1 showed the purity of RGCs was about 90%. The results of calcein-AM staining,which stained the living cells only,showed large cell body of RGCs and most of RGCs had a protuberance whose length was twice longer than the diameter of the cells. Conclusion RGCs cultured by serum-free medium has uniform size,good configuration,and high purity,which is adapt to the research of damage of RGCs caused by various factors and to evaluate the protective effects of neuroprotective agents.

Objective To establish the normal data of the fetal thorax,and to evaluate its values in the diagnosis of fetal thorax malformation.Methods Totally 398 normal singleton fetuses at 16 to 36 gestational weeks(GW) were enrolled,2D-US and 3D-US VOCAL technique were used to measure the 2D data and 3D volumes on the transverse section at the level of the four-chamber view,and the correlation among all measurements with GW was analyzed.Thirty fetuses collected randomly were examined to analyze the reliability.Nine fetuses with congenital thoracic dysplasia (CTD) and 10 fetuses with congenital diaphragmatic hernia (CDH) were assessed and compared with the normal fetuses.Results ① In healthy controls,the fetal thoracic 2D measurements and 3D volumes increased along with the growth of the GW.The regression equations were listed as follows:thoracic transverse diameter (cm) =-0.002 GW2 + 0.301 GW-1.510;thoracic anteroposterior diameter (cm) =0.003GW2 + 0.046GW + 0.666;thoracic area (cm2) =0.071GW2-1.466 GW + 14.728;thoracic circumference (cm) =0.01GW2 + 0.313GW + 3.341;thoracic volume (cm3) =0.285 GW2-7.797GW + 66.592;lung volume (cm3) =0.178 GW2-5.317GW + 45.539;the ratio of lung volume to thoracic volume =0.005GW + 0.396.② The reliabilities of the data obtained by the same/two different operators were good.③ CTD group was obviously lower than the healthy controls in all thoracic measurements (all P ＜0.01).There was no statistical difference in the 2D data between the CDH group and healthy controls (P ＞0.05),while the 3D volumes and the ratio of lung volume to thoracic volume were obviously lower than those in the healthy controls (P ＜0.01).Conclusions 2D-US can evaluate the fetal thoracic development and malformation preliminarily,but 3D-US VOCAL technology plays an important role in distinguishing different types of thoracic malformations.

Objective To investigate the intestinal absorption kinetics of breviscapine in rats.MethodsThe intestinal absorption in small intestine and colon of rats in situ was investigated using circular perfusion and HPLC methods.The in situ absorption of scutellarin in the small intestine was studied and the effects on the intestinal absorption were observed at the various concentrations and different pH values using the same methods.Results The results showed that there were significant differences in both absorptive percentage and absorption rate constant(ka)in the small intestine between the experimental groups of rats with and without bile duct being ligated.The absorption rate constants in small intestine and colon were(0.107 1?0.013 0)and(0.070 7?0.008 9)h-1,respectively.No saturation phenomenon occurred and ka was almost kept unchanged at different concentrations of breviscapine.The absorption of breviscapine was not influenced by pH values ranging from 6.0 to 7.4.Conclusion The results indicates that breviscapine is absorbed more in small intestine than in colon.The absorption of breviscapine complied with the passive diffusion mechanism and first order kinetics.In conclusion,these results suggest that breviscapine could be prepared in oral sustained-release dosage form.

0.05).But the P_ eff values were significantly increased in the presence of P-glycoportein(P-gp)inhibitor,verapamil or digoxin.CONCLUSION:Rhamnosylvitexin can be classified into high penetrating drug.Passive diffusion dominates the absorptive transport behavior of rhamnosylvitexin in HLF.There is not a preferential absorption zone in the intestine for rhamnosylvitexin.The absorption and secretion of rhamnosylvitexin in HLF are mediated by the efflux transport system,P-gp.

Objective To explore the change of function and expression of hematopoietic lineage cell specific protein-1 (HS1) and phosphorylated HS1 (p-HIS) and factors devoting to HS1 phosphorylation in platelet with sepsis.Methods Plasma with rich platelet was collected from 150 sepsis patients and 50 healthy subjects, and comparison of platelets adhesion and aggregation were detected by micro-pore method and platelet aggregation instrument.Meanwhile the ATP concentrations of washed platelet of two groups were detected by the kit to compare release reaction.And then total HS1 (t-HIS) and p-HS1 of platelet from two groups were compared by using western blot.Afterwards the specific inhibitors of Src and Syk were used to verify the HS1 activation regulated by Src and Syk in LPS-induced cell model.Results The significant differences were present between healthy subjects and sepsis patients in platelet counts, platelet distribution width (PDW) and mean platelet volume (MPV) (P ＜ 0.01).The data showed the sepsis patients had greater ability than healthy subjects in adhesion, aggregation and release reaction.Meanwhile the platelets of sepsis patients had higher concentration of t-HS1 and p-HS1 than healthy subjects, and the specific inhibitors of Src and Syk , PP2 and piceatannol, inhibited the increase in p-HS1 in LPS-induced cell model.Conclusions Function of platelet is closely related to HS1 in sepsis and it will be a target for sepsis therapy.

Objective To assess the clinical application values of HC visual laryngoscope in emergency tracheal intubation for severe craniocerebral trauma patients with basilar fracture.Methods Retrospective analysis was used to review 60 patients of severe traumatic brain injury with basilar fracture in emergency resuscitation room who needed the hospital emergency tracheal intubation.There were 30 patients operated with visual HC laryngoscope (the observation group), and the other 30 patients underwent common direct laryngoscope (the control group).The index of the degree of exposure,intubation time,success rate of intubation were compared between the two groups.ResultsThere were 28 cases (93.33%) of Cormark-Lehane grading Ⅰ to Ⅱ in observation group,which was significantly higher than 20 cases (66.67%) in the control group(P< 0.05).The average intubation time in observation group was (34.3±6.7) seconds,and it was shorter than (44.5±5.7) seconds in the control group (P<0.05).In observation group,26 cases (86.67%) were of one-time successful intubation,which was higher than 19 cases (63.33%) in the control group,and the difference was statistically significant(P<0.05).ConclusionHC visual laryngoscope in emergency tracheal intubation for severe craniocerebral trauma patients with basilar fracture can improve the success rate of intubation,shorten the intubation time,reduce adverse reactions,and it is worthy of clinical application.

Objective:To investigate the role of TRPV5 and TRPV6 in intracellular calcium regulation and biological behaviors of SW480 colon cancer cells. Methods:qRT-PCR, Western blot, and immunocytochemistry were applied to determine the mRNA and protein ex-pression levels of TRPV5 and TRPV6 in SW480 colon cancer cell line upon treatment with TRPV5 and TRPV6 agonist, 1-25(OH)2D3, and inhibitor, CuCl2. The change of intracellular Ca2+level was examined with a confocal laser scanning microscope. Scratch test, MTT, and TUNEL assays were used to analyze the cell migration, proliferation, and apoptosis, respectively. Results:As an agonist of TRPV5 and TRPV6, 1-25(OH)2D3 significantly up-regulated the mRNA and protein expression levels of TRPV5 and TRPV6 in SW480 cell lines. On the other hand, CuCl2, being an inhibitor of TRPV5 and TRPV6, effectively down-regulated the TRPV5 and TRPV6 mRNA and protein expres-sion levels (P<0.05). The intracellular calcium concentration in SW480 cell line significantly increased upon treatment with 1-25 (OH)2D3, and significantly decreased with CuCl2 treatment (P<0.05). 1-25(OH)2D3 promoted cell proliferation and migration, and inhibit-ed apoptosis of SW480 cell in a time-and dose-dependent manner (P<0.05). However, CuCl2 significantly repressed cell proliferation and migration and induced apoptosis (P<0.05). Conclusion: TRPV5 and TRPV6 can affect the biological behaviors of colon cancer SW480 cells by regulating intracellular Ca2+level.

Objective To investigate the levels of IL-6 in serum and gastric mucosa of servicemen before and after mili-tary maneuver and their significance .Methods The level of serum IL-6 in 60 servicemen was studied by enzyme-linked immunosorbent assay .The expression of IL-6 in gastric mucosa from the 60 servicemen was detected by immunochemical staining.Results The level of IL-6 in serum of servicemen after military maneuver was significantly higher than that before military maneuver (P<0.01).IL-6 was found to be expressed in the cytoplasm of epithelioglandular cells and also in the cytoplasm of mesenchymal cells .The expression of IL-6 in gastirc mucosa was significantly stronger after military maneuver than before military maneuver (P<0.01).Conclusion IL-6 might be involved in the development of pathological changes in gastric mucosa caused by military stress in troops .Examination of the level of IL-6 in the early stage of military stress might help assess the chance of gastric mucosa lesion and its severity .

Objective To examine loss of heterozygosity (LOH) and microsatellite instability (MSI) of locus D8S532 on chromosome 8 and their influence on the expression of sFRP1 in the hepatocellular carcinoma (HCCs), which may provide an experimental evidence for clarifying the mechanism of sFRP1 gene and tumor development. Methods DNA was extracted from formalin-fixed paraffin-embedded tissues. Polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) and ordinary silver stain were used to study LOH and MSI of locus D8S532. Envision immunohistochemistry, Leica-Qwin computerized imaging system and Image-Pro PluS (IPP) version 4.5 professional imaging analysis software were used to assess the expression of sFRP1. Results The detection rates of LOH and MSI of locus D8S532 in the 36 specimens of HCC were 11.11% and 8.33% respectively. The down-regulation of sFRP1 was observed in 31 of 36 HCCs (86.11%) compared with non-carcinoma liver tissues, and the positive rate of sFRP1 protein of the HCCs was 52.78%( 19/36 ). The frequency of LOH was lower in the cases with positive expression of sFRP1 protein than those negative (0 vs 23.53%, P <0.05). Conclusion It was a common phenomenon that expression of sFRP1 protein is negative or low in Chinese with HCCs. The genetic instability of sFRP1 gene was one of causes, which lead to HCCs. LOH may play a major role in negative expression of sFRP1.

Objective To investigate the effect and mechanism of cell differentiation agent Ⅱ (CDA-Ⅱ) on the differentiation of human acute mycloid leukemia (APL) HL-60 cells.Methods The cell morphology and differentiation was detected by Wright-Giemsa staining,the expression of cell surface differentiation antigen CD11b of HL-60 was detected by flow cytometry,the differentially expressed genes were screened by gene expression profile chip (NimbleGen).Results The result of Wright-Giemsa staining showed that CDA-Ⅱ induced HL-60 differentiation towards mature stages in a time-dependent manner.After treated with CDA-Ⅱ,the percentage of CD11b-positive HL-60 cells significantly increased in a time-and dose-dependent manner.The result of gene expression profiling indicated differentially expressed genes including 113 up-regulation genes and 140 down-regulation genes.The up-regulation expression genes involved in six pathways including mineral absorption,complement and coagulation cascades,down-regulation expression genes involved in nine pathways including pyrimidine metabolism,RNA polymerase,purine metabolism and so on.Conclusion CDA-Ⅱ can induce HL-60 differentiation and make gene differentially expressed.The data provide the feasibility of CDA-Ⅱ in differentiation induction therapy for APL.