AIM: To optimize and establish the extraction techn ology of water-soluble polysaccharides from Ulva pertusa. TLC was employed to determine the mainly neutral sugars in the Ulvan. METHODS: The orthogonal test was employed to test the effects of th e four factors including volume of water, temperature of extraction, time of ext raction and pH of water on the yield and total carbohydrate contents of water-s oluble polysaccharides from Ulva pertusa and optimize the extraction technol ogy. Five groups of parallel tests were carried out by the optimum parameters es tablished and the sugar constituents of products were analyzed by TLC. RESULTS: Temperature of extraction and time of extraction have sign ificant effects on the extraction yield of water-soluble polysaccharides from Ulva pertu sa, but, for total carbohydrate content, only temperature of extraction was a significant factor. The mainly neutral sugars were composed of rhamnose, xylose and glucose. CONCLUSION: The optimum extraction technology was A 1B 3C 3D 3 when the yield and total carbohydrate content were considered as preferential pa rameters.

Objective To investigate relationship between levels of follicle-sitimulating horomone receptor(FSHR) and ovarian response induced by gonadotropin hormone,and whether the FSHR expression is correlated with in vitro fertilization(IVF) outcome.Methods Granulose cells were collected from 43 women receiving IVF-embryo transplantation(IVF-ET).According to the number of oocyte,the women were divided into three groups: low response(15).The expression intensity of FSHR was measured by immunohistochemistry technique.The expression intensity of FSHR on the granular cell,the embryological and clinical outcomes were compared and analyzed. Results The expression of FSHR was significantly different in three groups with the highest in high response group(P0.05).The FSHR level was positively correlated either with the number of oocyte (r=0.719) or with the serum E2 levels(r=0.516,P0.05). Conclusion Ovarian response to gonadotropin hormone stimulation is correlated with the level of FSHR in the granulose cells.The development of follicles may be influenced by it.

Humanin(HN,its analogue [Gly14]-Humanin,HNG)was originally identified as an endogenous peptide that protects neuronal cells from apoptosis induced by various types of Alzheimer's disease-related insults.But the relative low content of this peptide in its natural sources limits its further characterization.An expression vector pET32a/HNG was corstructed and transformed it into E.coli BL21 trxB(DE3).HNG was expressed as a fusion protein in the soluble fraction and was purified by nickel affinity chromatography.Subsequently,the purified fusion protein was cleaved by enterokinase and was further purified by reverse-phase HPLC.A 23 mg recombinant HNG(rHNG)from 1 L bacterial culture was purified.The molecular weight of rHNG determined by ESI-MS was 2876.5 Da which was the expected size for correctly processed peptide.The N-terminal amino acid sequence of rHNG determined by Edman degradation method is identical to the theoretical sequence.Neuroprotective bioassay studies of rHNG exhibited its potential neuroprotective effect comparable to that of the natural HNG peptide.

· AIM: To explore the relationship between the expression of Fas/FasL and the apoptosis in retinal ischemia/reperfusion injury of rats, as well as the therapeutic effects of basic fibroblast growth factor (bFGF)on the ischemic retina.injury were made by transiently elevating introcular pressure. A total of 28 rats were divided into Normal Group and Operative Group. The latter were subdivided into 1, 6, 12, 24, 48 and 72h after reperfusion, in which the left eyes of the rats were in the ischemia/reperfusion groups and the right ones were in the treatment groups (bFGF intracameral injection). Apoptosis was assessed by the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labelling (TUNEL) method, and theexpression of Fas/FasL ligand was studied by strept avidin-biotin complex (SABC) immunohistochemistry.mai rats' retinae, but there were a significant number of TUNEL positive cells in 6-24h after transient ischemia followed by a decrease at 48h. The number of TUNEL positive cells reached a maximum at 24h after ischemia.The expression of Fas gradually increased as early as at 6h, reached a peak at 24h, then decreased at 48h. Similarly, the expression of Fas ligand was at peak in 24-48h in GCL and INL of retina. bFGF ministered before reperfusion inhibited apoptotsis and ameliorated the tissue damage. It also diminished Fas and FasL expression in ischemic/reperfused retina.siently elevated IOP induced apoptosis of cells in the retina. Fas/FasL may have an important role in the early events of the apoptotic pathways. bFGF can rescue RGCs from retinal ischemia/reperfusion injury through down-regulation of Fas and Fas ligand expression and may represent an important mechanism for therapeutic neuroprotection.

Objective To observe the effect of external use of Curcuma oil on the hyperplasia of mammary glands in rats and explore the mechanism. Methods Sixty female Wistar rats were randomly divided into following groups:normal con-trol group, model control group, Sanjierubi plaster group, low-, medium- and high-dose curcuma oil emulsion groups ( n=10 each).The models of hyperplasia of mammary glands were established by intramuscular injection of estradiol benzoate and proges-terone into the medial part of the rat hind limb.Different doses of medicines were given for 4 consecutive weeks.Enzyme-linked im-munosorbent assay, HE staining and immunohistochemistry were used to investigate the action mechanism of curcuma oil emulsion against mammary gland hyperplasia. Results Curcuma oil emulsion had preventive and therapeutic effects on the hyperplasia of mammary glands.The diameter of breasts was significantly reduced, the body weight restored, serum estradiol, follicle-stimulating hormone and prolactin levels profoundly decreased, progesterone, testosterone and luteinizing hormone levels markedly increased and the number and diameter of lobular acini obviously reduced in high-dose curcuma oil emulsion group when compared with those in model control group (P<0.05 or P<0.01). Conclusion Curcuma oil emulsion can remarkably improve the disturb-ance of serum hormones and inhibit the occurrence of hyperplasia of mammary glands.

Objective To optimize the extraction process of anti-hepatic fibrosis water-soluble component from Carthamus tinctorius L. via the orthogonal method. Methods The influences of type and content of solvent,extraction time,and extraction frequency on processing were investigated by assessing the content of hydroxysafflor yellow A,yield of extraction and anti-hepatic fibrosis potency via the high performance liquid chromatography and enzyme-linked immunosorbent assay by L9(3)4 orthogonal test. Results The optimum extracting condition for the anti-hepatic fibrosis water-soluble component from carthamus tinctorius L. was as follows: extracting with 10 times the volume of 10% ethanol for twice, 0. 5 h in each. Conclusion The process is scientific and feasible, which serves as a guideline for the production of the anti-hepatic fibrosis water-soluble components.

Objective To prepare the hydroxysafflor yellow A (HSY-A)enteric coating pellets,and investigate in vitro release and in vivo pharmacokinetic characteristics.Methods HSY-A enteric coating pellets were prepared by extrude-rounding and fluid bed technique.The micromeritic characteristics,the factors affecting the release properties of enteric coating pellets the release mechanism were explored,and the in vivo pharmacokinetic behaviors were also evaluated.Results The in vitro release behavior of HSY-A from enteric coating pellets could be described by Ritger-peppas equation,and fit diffusion mechanism,in vivo pharmacokinetic test confirmed the argument that pellets have good acid residence and enteric properties. Conclusion HSY-A enteric coating pellets have been successfully prepared and the expected release properties is achieved in the study.

Objective To investigate the pathological and immunohistochemical characteristics of female bladder ectopic skene glands.Methods A female with bladder tumor was treated in our hospital in May 2013.Preoperative so(n)graphy revealed a 0.9 cm×0.6 cm round solid mass in the bottom of bladder wall.Mass was hypoechoic homogeneous with regular shape,blood flow within the mass was noted.The tumor was treated with transurethral resection.Routine pathological examination suggested bladder ectopic Skene glands.Immunohistochemical stains for prostate specific antigen (PSA),prostate spectific acid phosphatase (PSAP),androgen receptor (AR),estrogen receptor (ER),CD10,cytokeratin 14 (CK14),cytokeratin 18 (CK18),P63,high molecular weight cytokeratin (34βE12),α-methylacyl-CoA racemase (AMACR/p504s) were further performed.Results Routine pathological examination showed prostate glands composed of prostate gland epithelial cells and basal cells in a submucosal location.Immunohistochemical stains showed:PSA-,PSAP +,AR +,ER-,CD10+,CK18 +,CK14-,P63 +,34βE12 +,AMACR-.Conclusions Routine pathological examination combined with immunohistochemical stains such as PSA,PSAP,and others,can be used to diagnose ectopic Skene glands disease.Female bladder ectopic Skene glands is a benign lesion,and the prognosis is good.

Objective To explore the changes of soluble tumor necrosis factor receptor type 1(sTNFR1)in children with acute virus encephalitis(VE)and its clinical significance.Methods The levels of tumor necrosis factor-?(TNF-?),sTNFR1 and neuron specific enolase(NSE)in cerebrospinal fluid were determined by enzyme-linked immunosorbent assay(ELISA)in 55 children who were admitted with VE,including 25 cases with severe VE(SVE)and 30 cases with mild VE(MVE).Fifteen cases without VE were as control group.Results The levels of TNF-?,sTNFR1 in cerebrospinal fluid in encephalitis children were significantly higher than that in control group(Pa

Objective To investigate the function of transferrin-DNA complex,transported by transferrin(Tf)and trans-arterial injection via interventional approach be the duel-target-orientated delivery and the transferring into malignant cells to get more effective therapy.Methods p53-LipofectAMINE ligand with different concentrations of Tf(0,10,25,50,100?g)transfected the 4 strains including LM6、Hep3B、YY and L02 in vitro to evaluate the gene transfeetion efficiency through western blot.Then,after setting up the VX2 hepatocarcinoma models,we delivered the Tf-p53-LipofeetAMINE complex into the hepatic arteries via interventional techniques to analyse the transfection efficiency in vivo.Results Tf,within the range of 10 100?g,could increase gene transfection efficiency mediated by liposome,and the efficiency increases with the raise of Tf concentration.Combination with interventional technique to inject Tf-DNA complex into tumor arteries,gene transfeetion efficiency was enhanced in rabbit models.Conclusion Tf can enhance gene- liposome transfection efficiency,furthermore with combination of interventional catheter technique,there would be a potential duel-target-orientated gene therapy method.(J Intervent Radiol,2007,16:99-103)