Purpose　To study the proteins expression of genes related to apoptosis of retinal cells in development of human fetus．　Methods　Fifty cases of retinas of human fetus aged from 12 to 38 weeks were collected and paraffin embedded sections were made． Immunohistochemical method was used．　Results　Fas protein was expressed by cells of ganglion cell layer, inner and outer nuclear later, which were just formed on 16th week． It was not expressed until 38th week, Fas(+) staining appeared in layers of retina． Fas-L(+) staining was detected in cells of layers of retina on 26th week and the positive staining located in ganglion cell layer on 32th week． Neuronal fiber layer was Fas-L positive． Bax positive staining was detected on 8th week． Bax positive nucleus were observed mainly in GCL and ONL on 16th week． It was in INL on 24th week and in Müller cells inner terminates on 26th week． After this time, all cells of retina were bax immune negative staining． Bcl-2(+) staining appeared in differentiating neuroblastic layer on 16th week． Beginning on 24th week, bcl-2 (+) staining was observed in glial cells of GCL and inner terminates of Müller cell．　Conclusion　Apoptosis of developing retinal cell may be Fas/Fas-L independent and bax may be involved in apoptosis of the cells．

Objective To explore the mechanism of QHR (main ingredients:Gypsum, Anemarrhema Asphodeloides Bge, Paeonia Lactiflora Pall) on secretive function of vaso-endothelial cells in rabbits with of endotoxemia. Methods Endotoxemia model was induced by colibacillary endotoxin via auricular intravenous injection in rabbits. The changes on pathomorphology and secretive function of vaso-endothelial cells in the model and QHR group were observed. Results It showed the existence of microthrombus in the lung via microscope in the model. The balance destruction of t-PA/PAI, NO/ET, TXB2/6-K-PGF1? in the model (P

Objective　To study the distribution of insulin-like Growth Factors (IGFs) in liver、 spleen、kidney、thymus and supraadrenal glands of fetus from 16 to 24 weeks.Method　Using ABC immunohistochemistry. Results　 IGF-I positive cells were found in these organs during this period of fetal life, but the differences of number and reaction intensity of IGF-I positive cells appeared in different organs and individuals.Conclusion　These organs tissues can express IGF-I during fetal development and play paracrine and/or autocrine role in cellular proliferation and differentiation of these fetal tissue.

Objective To observe the expression of p53, bcl 2 genes, vascular endothelial cell growth factor (VEGF), basic fibroblast growth factor (bFGF), insulin like growth factor I (IGF I), and the receptors of these factors of retinal vascular endothelial cells (VECs) of 1 to 20 week diabetic rats, and the relationship between the expressions and cell cycle arrest. Methods Retinal sections of diabetic rats induced by alloxan were immunohistochemically stained and observed by light microscopy (LM) and electron microscopy (EM). Dot blotting and Western blotting were used to determine the expression of mRNA, proteins of p53 and bcl 2. Results Under LM, immunohistochemical positive expression of p53 and bcl 2 were found on the vessels of ganglion cell layer and inner nuclear layer of retinae of 8 to 20 week diabetic rats; under EM, these substances were observed depositing in VECs. The retinal VECs also expressed VEGF, bFGF, IGF I and their receptors. There was no positive expression of other cell types in these retinae, all cell types of retinae in control group, or all cells of retinae of diabetic rats with the course of disease of 1 to 6 weeks. The result of dot blotting revealed that retinal tissue of 20 week diabetic rat expressed p53 and bcl 2 mRNA, and the result of Western blotting revealed that they also expressed p53 and bcl 2 proteins. But retinal tissues of control group did not. Positive expression of bax was not found in the retinae in control group or 1 to 20 week diabetic rats. Conclusion p53, bcl 2 may introduce cell cycle arrest of VECs of retinae in 8 to 20 week diabetic rats. High glucose might stimulate the expression of VEGF, bFGF, IGF I and their receptors, and the growth factors may keep VECs surviving by self secretion.

A selective precolumn derivatization liquid chromatography-tandem mass spectrometric (LC-MS/MS) method for the determination of glucosamine in human plasma and urine has been developed and validated.Glucosamine was derivatized by o-phthalaldehyde/3-mercaptopropionic acid.Chromatographic separation was performed on a Phenomenex ODS column (150 mm × 4.6mm,5μm) using linear gradient elution by a mobile phase consisting of methanol (A),and an aqueous solution containing 0.2％ ammonium acetate and 0.1％ formic acid (B) at a flow rate of 1 mL/min.Tolterodine tartrate was used as the internal standard (IS).With protein precipitation by acetonitrile and then the simple one-step derivatization,a sensitive bio-assay was achieved with the lower limit of quantitation (LLOQ) as low as 12 ng/mL for plasma.The standard addition calibration curves suitable for clinical sample analysis showed good linearity over the range of 0.012-8.27 μg/mL in plasma and 1.80-84.1 μg/mL in urine.The fully validated method has been successfully applied to a pharmacokinetic study of compound glucosamine sulfate dispersible tablets in health Chinese volunteers receiving single oral doses at 500,1000 and 1500 mg of glucosamine sulfate,as well as multiple oral doses of 500 mg t.i.d.for 7 consecutive days.

Aim To study the pharmacokinetics characteristics of galanthamine hydrohromide sustained release tablets and conventional tablets in healthy volunteers after a single and multiple oral doses. MethodsA single and multiple oral doses of galanthamine hydrohromide sustained release tablets and conventional tablets were given to 20 healthy male volunteers in a randomized cross-over study. We developed an LC-MS assay using naloxone as the internal standard to determine the plasma concentrations of galanthamine, calculate the pharmacokinetic parameters and evaluate the relative bioavailability and sustained release characteristics of galanthamine hydrohromide sustained release tablet. Results The pharmacokinetic parameters of the sustained release tablet and conventional tablet obtained from the single-dose study were as follows: the HVD_12 C_max(time span during which the plasma concentration is at least half of the C_max value)were (15.4?1.7) h and (5.4?2.5) h, the retard quotients (R△,the HVD_12 C_max ratio of sustained release tablets to conventional tablets) of sustained release tablet was 3.4?1.4, the T_max were (4.4?1.5) h and (1.3?1.2) h, the C_max were (27.5?2.9) ?g?L-1 and (53.7?12.7) ?g?L-1.Results showed significant sustained release characteristics of the sustained release tablet. The relative bioavailability of the sustained release tablet was (95.9?14.2) %。The pharmacokinetic parameters of the sustained release tablet and conventional tablet obtained from the multi-dose study were as follows: the T_max were (3.0?1.6) h and (0.9?0.3) h,the CSS_max were (58.8?9.4) ?g?L-1 and (52.0?6.9) ?g?L-1,the CSS_min were (16.2?4.0) ?g?L-1 and (22.5?5.0) ?g?L-1,the C_av were (39.0?3.9) ?g?L-1 and (37.1?5.0) ?g?L-1,the DF were 1.1 ?0.3 and 0.8?0.1, respectively. Results of two one-side t test showed that AUC_SS、CSS_max、C_av of two tablets were bioeqivalent. Conclusion Results showed that the sustained release tablet and the regular tablet were bioequivalent in absorbed extent, and the sustained release tablet exhibited a good retarding effect in release.

Aim To research the protective effects of specnuezhenide, an active compound of traditional Chinese medicine, against CCl4-induced hepatic dam-age in mice. Methods The model of acute hepatic damage caused by CCl4 intraperitoneal injection on mice was obtained. The levels of ALT, AST, TNF-α, IL-1β, and IL-6 in serum, the activities of MDA, SOD, and GSH-Px in liver homogenates, and the his-topathological organization of liver sections were also examined to observe the protective effects of specnu-ezhenide. Results Specnuezhenide markedly de-creased the CCl4-induced elevation of serum ALT and AST activities, and improved hepatic histopathology changes. Specnuezhenide also significantly decreased the content of MDA in liver tissues, meanwhile in-creased the activities of antioxidant enzymes SOD and GSH-Px. In addition, specnuezhenide reduced the lev-els of TNF-α, IL-1β and IL-6 . Conclusions Specnu-ezhenide has significant protective effects against CCl4-induced hepatic damage in mice, and the possible un-derlying mechanisms of the activity could be due to its antioxidant and anti-inflammatory effects.

OBJECTIVE:To prepare famotidine sodium chloride injection METHODS:The famotidine sodium chloride injection was prepared with aspartic acid as solutizing agent and sodium chloride injection as solvent The contents of famotidine and its related substances were determined by means of HPLC Influencing factor and the stability of famotidine were studied and the therapeutic effect was observed RESULTS:The content of famotidine was slowly reduced while temperature rising and time prolongine The content of famotidine was 98 6% at the end of the 6th month at 40℃ with accelerating experiment Its content was 98 4% after one year storage under room temperature Its content was 98 5% at the 10th day after illumination with 4 500Lx light in accelerating experiment The content of related substances was less than 2% The total effective rate was 93 75% CONCLUSION:Famotidine sodium chloride injection was stable in property and had definite therapeutic effect,and its clinical application was simple and free from contamination

Objective To investigate the changes of Treg cells,Th17 cells and related cytokines in pe?ripheral blood of patients with severe hand?foot?mouth disease( HFMD) in the initial stage of infection caused by Enterovirus 71 virus. Methods Thirty?five cases of severe HFMD children in the Fifth Hospital of Shijiazhuang from May 2013 to December 2013 were selected as the research subjects,including 27 cases of severe formed group and 8 cases of danger severe formed group. The frequencies of Th17 cells ( CD3+CD8?IL?17+) and Treg cells( CD4+CD25+Foxp3+) in peripheral blood were detected by flow cytometry in patients with severe HFMD and 31 healthy children(control group). Moreover the serum levels of interleukin 6(IL?6),interleukin 17(IL?17) ,interleukin 10( IL?10) ,interleukin 23( IL?23) ,and human transforming growth factorβ1 ( TGF?β1) were measured by ELISA. Results As compared with those in control group,the frequencies of Treg cells,ratio of Treg/Th17,serum levels of TGF?β1, IL?10 in severe HFMD groups were significantly decreased ( F=23. 23, 30. 88,10. 766,11. 680 respectively;P<0. 05) . However the frequency of Th17 cells,serum levels of IL?6,IL?17,IL?23 in severe HFMD groups were significantly increased( F=646. 81,5. 904,10. 557,3. 490 respectively;P<0. 05) . Conclusion The changes of Treg and Thl7 cells and related cytokines appear in peripheral blood of patients with HFMD in the initial stage of infection caused by Enterovirus 71 virus. The frequency of Treg cell is decreased,however,the frequency of Thl7 cells frequency is increased,as a result,which may spread inflammato?ry reaction and cause complication.

Objective To investigate the effect of hydrogen on radiation-induced acute injury in rat brain.Methods Forty-five mature Sprague-Dawley rats were randomly divided into three groups:saline therapy group,hydrogen therapy group and healthy control group.The whole brain of SD rat was irradiated with single dose of 20 Gy by 4 MeV electrons.Rats in therapy group were injected with hydrogen-rich saline after irradiation and were sacrificed at 1,3,7,14 d post-irradiation.The changes of malonaldehyde (MDA),superoxidase dismutase (SOD) and 8-hydroxydeoxygunosine (8-OHdG) in brain homogenate and the pathological changes in brain hippocampus were observed.Results The brain water content (t=3.78,3.18,P＜0.05) and the contents of 8-OHdG (t=2.33,2.71,2.33,P＜0.05) in the therapy group was lower than the control group at 7 d and 14 d post-irradiation.The contents of SOD were significantly higher(t =2.41-2.92,P ＜ 0.05) from 1 to 7 day,while the contents of MDA were significantly lower in therapy group than those in the control group from 1 to 14 day post-irradiation (t =4.01-6.20,P ＜ 0.05).Moreover,the damage degree in the nerve cells of hippocampus was less compared to the control group.Conclusions The hydrogen-rich saline could have protection role in irradiation-induced acute brain injury in rats.