Mucins(MUC)are secreted by the epithelial cells of polymer ,the height of glycosylation gly-coprotein,widely existing in the respiratory system ,digestive system,urogenital system in the mucosa epithelium and mucus secretion .The resent studies show that sticks are closely associated with tumor protein family .Among them,the sticky protein 1(CA153)and 16 mucins(CA125)has been confirmed that the abnormal expression in the bile duct carcinoma.However,there is unclear relationship between the development ,invasion,metastasis and prognosis of gallbladder .Based on the retrospective literature at home and abroad in recent years , the research progress on the sticky protein 1 and mucins 16 in the gallbladder is summarized in the present review .

Objective To observe the long-term effect of suramin on the inhibition of proliferation of human retinal pigment epithelial (RPE) cells in vitro. Methods RPE cells grown in 9 pieces of 96-well plate (12 wells each plate) were divided into experimental and control group, with 6 wells in each group. The concentration of 0.1 ml RPE cells in each well is 5?104 cells/ml. After the change of the medium, RPE cells were treated with suramin (250 ?g/ml) in experimental group while treated with nothing in the control group. The medium of the 2 groups were changed to the normal medium after 4 days. At the 1~st , 2~nd , and 4~th day after the addition of suramin and at the 1~st , 2~nd , 3~rd , 5~th , 6~th , 7~th , 9~th , 11~th and 13~th day after removing suramin, 1 plate was randomly selected to stop culturing, and the proliferation of RPE cells were detected by methyl thiazolyl tetrazolium (MTT) assay. Results Under reversed microscope, RPE cells in control group were fused completely at the 7~th day after inoculation. The extracellular space of RPE cells in experimental groups was larger than that in the control group, and remained unfused at the 13~th day after inoculation. The inhibitory rate of proliferation of RPE cells at the first day after treated with suramin was 14.85% and increased to the highest 25.79% at the 4~th day. The first day after the suramin-containing media was removed, the inhibitory rate decreased to 12.35%, and then raised gradually to over 20% at the 3~rd to 5~th day. Finally, the rate drop to 14.71%. Conclusion Suramin has the long-term effect on the inhibition of RPE cells induced by serum, especially the inhibitive effect after the remove of suramin, which indicates the specific double-peak inhibition during the whole process.

Objective To observe the ultrastructural characteristics of human retinal progenitor cells cultured in vitro. Methods Six 5-month-old human fetuses(12 eyes)without eye diseases were selected. Retinal progenitor cells from the retina of one eye of each fetus were cultured in vitro,and observed by transmission electronic microscopy(TEM); while those from the other eye were directly observed by TEM. Results Abundant heterochromatin were found in the karyon of 5-month embryonic retinal neuroepithelial cells,and the figure of the karyons was irregular.A few scattered initial cells were seen in retinal neuroepithelial layer with large karyon,smooth surface,abundant euchromatin,and distinct nucleolus.The human retinal progenitor cells cultured in vitro had the same ultrastructural characteristics as the initial cells:with huge karyon which almost occupied the whole cell,little cytoplasm,distint nucleolus,abundant euchromatin,and little heterochromatin.The cells clung to each other in the neural globoid cell mass.The size of the outer cells was large,and karyokinesis could be found. Conclusion The cultured human retinal progenitor cells are provided with the same ultrastructure characteristics as the initial cells.

Objective:To find small molecular leads for inhibition on early stage of HIV infection by identification and characterization of the HIV-1 gp41 C-helix mimotopes.Methods:For identification of the gp41 C-helix mimotopes,C7C phage display peptide library was biopanning by using a synthetic peptide N36 which was derived from the gp41 N-helix as target.After three rounds of screening,positive phage clones were identified by ELISA and sequenced.Results:16 of 26 phage clones were identified to bind with peptide N36,and 10 of them were sequenced.Every clone of ten clones contains at least two hydrophobic residues,which may dock into the hydrophobic pocket in the gp41 N-helix domain.9 of the 10 clones have a conservative sequence WW,which may mimic the W628 and W631 in C-helix to interact with the hydrophobic residues in the gp41 pocket.One clone expressing the conservative sequence named clone No.8(CYWWHRLHC) was selected for characterization.The binding between the clone No.8 and N36 was blocked by free peptide N36.And the binding between clone No.8 and peptide N36 was inhibited by peptide C34(IC 50=12.5 ?g/ml).Conclusion:The short circular peptides displayed on phages containing WW residues may mimic the conformational epitope of the HIV-1 gp41 C-helix to interact with the N-helix.This information may be useful for design of HIV-1 fusion inhibitors.

AIM: To observe the effect of ginkgolide B (CB) on the intracellular calcium ion concentration ( [ Ca~(2+) ]_i) and mitochondrial function of cultured rat retinal neurons in vitro. METHODS: in vitro primary culture of rat retinal neurons was used in the experiment. The apoptosis model of glutamate - induced retinal neurons was established and co - cultured with ginkgolide B. The [ Ca~(2+) ]_i and mitochondrial membrane potential of the retinal neurons were detected by laser scanning confocal microscope. RESULTS: Glutamate decreased the survival rate of retinal neurons, increased the apoptosis and the [ Ca~(2+) ]_i, lowered the mitochondrial membrane potential. The [ Ca~(2+) ]_i was clearly diminished and the mitochondrial membrane potential was significantly increased with the GB intervention, and the apoptosis decreased significantly. CONCLUSION: GB protects retinal neurons from glutamate induced neurotoxicity. The effect of GB on retinal neurons might be due to its ability to decrease the [Ca~(2+) ]_i and increase mitochondrial membrane potential.

Objective To investigate the effect of recombinant plasmid pshRNA-DNMT3b on expression of DNMT3b mRNA and protein and on the proliferation of bladder cancer T24 cells,and research the function of DNMT3b in the process of bladder tumor formation.Methods There were three groups in this study,which are blank controller,HK and pshRNA-DNMT3b(24h,48h,72h),respectively.T24 cells were cultured routinely and transfected by the recombinant plasmids with lipfectamine 2000.The cells were detected by methods of RT-PCR,western blot and MTT.The varying level of DNMT3b mRNA and expression protein,and the conditions of cellular survival rate were observed.Results The recombinant plasmids were successfully transfected into T24 cell lines.The grey valHe of RT-PCR elctrophoretogram was analyzed by the software of Gel-pro analyzer,the rate of blank controller,HK and pshRNA-DNMT3b(24h,48h,72h),was (99.56±1.24)%,(99.12±1.35)%,(75.77±1.42)%,(44.69±1.05)%and(20.52±0.89)%,respectively.The analytical resuit of western blot image was(99.43±1.28)%,(98.90±1.31)%,(67.83±1.02)%,(43.43±1.05)%and(21.92±0.89)%.There was no statistically difference in survival between blank control and HK(P＞0.05).The group of pshRNA-DNMT3b and other two groups had statistical difference only at the 72th hour and the cell inhibitory growth rate only increase 0.45%.Conclusions The recombinant ptasmid pshRNA-DNMT3b can inhibit the expression of mRNA and protein of DNMT3b effectively.However,it has slight function on inhibiting cell proliferation.

Objective To analyze the clinical features and risk factors for mortality of patients with severe fever with thrombocytopenia syndrome (SFTS) in Zhoushan, the eastern coastal of China with high incidence of severe fever with thrambocytopenia syndrome bunyavirus infection, to provide reference for reducing the mortality rate of SFTS.Methods Clinical data of 107 cases of SFTS from Zhoushan Hospital during June 2011 to June 2016 were retrospectively analyzed.According to the prognosis, patients were divided into survival group and death group.The clinical features and the laboratory results were analyzed with a case-control method to analyze the prognostic factors.Normal distribution data were compared with the independent t test.Kolmogorov-Smirnov Z test were used in data with skewness distribution.Categorical data were analyze by chi-square test.The related risk factors were analyzed with the receiver-operating characteristic (ROC) curve and multivariate unconditioned logistics regression analysis.Results Seventeen cases among 107 STFs patients died, yielding the mortality rate of 15.9%.The proportion of patients suffering from two or more underlying diseases, with disorders of consciousness, activated partial thromboplastin time (APTT), the level of creatine kinase (CK), lactate dehydrogenase (LDH) as well as sepsis-related or sequential organ failure assessment (SOFA) score in death group were all significantly higher than those in the survival group (all P<0.05).The Ca2+ level and fibrinogen level in death group were significantly lower than those in the survival group (both P<0.05).Indexes mentioned above were analyzed by ROC curve, and the calculated cut-off value was set as the optimal diagnostic thresholds.These data were then included into the multivariate logistic regression analysis.It turned out that Ca2+<1.625 mmol/L, APTT >73.45 s, SOFA scores >9 were the independent risk factors for mortality of SFTS (OR=6.947, 8.459 and 11.770, respectively, all P<0.05).Conclusion Ca2+, APTT and SOFA score are the independent risk factors for prognosis of SFTS, which provide reference for prognostic evaluation of SFTS.

AIM: To observe the effect of ginkgolide B (GB) on glutamate-induced apoptosis in the cultured neurons of rat retina. METHODS: Neurons of rat retina were cultured and apoptosis was induced by glutamate. The neurons were cultured with different concentration of GB and the survival rate was monitored by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. The apoptosis in the cultured neurons and the expression of Bcl-2 and Bax were observed by flow cytometry. RESULTS: After exposed to glutamate, the survival rate and the number of Bcl-2 positive cells obviously decreased. At the same time, the number of Bax positive cells obviously increased, and the number of the apoptotic cells also obviously increased. Such phenomena were relieved by the treatment of ginkgolide B, with raise of survival rate and the expression of Bcl-2. Meanwhile, the expression of Bax and the apoptosis of neurocytes obviously decreased. CONCLUSIONS: Ginkgolide B protects retinal neurons from the virulence induced by glutamate. Such effects of GB might be brought about by increasing the expression of Bcl-2 while decreasing Bax, resulting in increasing the ratio of Bcl-2 to Bax and so reducing the apoptosis in the cultured neurons of rat retina.

AIM:To observe the effect of ginkgolide B (GB) on the intracellular calcium ion concentration ([Ca2+]i) and mitochondrial function of cultured rat retinal neurons in vitro.METHODS:in vitro primary culture of rat retinal neurons was used in the experiment. The apoptosis model of glutamate-induced retinal neurons was established and co-cultured with ginkgolide B. The [Ca2+]i and mitochondrial membrane potential of the retinal neurons were detected by laser scanning confocal microscope.RESULTS:Glutamate decreased the survival rate of retinal neurons,increased the apoptosis and the [Ca2+]i,lowered the mitochondrial membrane potential. The [Ca2+]i was clearly diminished and the mitochondrial membrane potential was significantly increased with the GB intervention,and the apoptosis decreased significantly.CONCLUSION:GB protects retinal neurons from glutamate induced neurotoxicity. The effect of GB on retinal neurons might be due to its ability to decrease the [Ca2+]i and increase mitochondrial membrane potential.

OBJECTIVETo prepare ginkgolide B-loaded self microemulsifying drug delivery system (SMEDDS) and evaluate its quality.

METHODThe solubility of ginkgolide B in different oil, surfactant and co-surfactant were measured by HPLC-ESI-MS. The GB-SMEDDS formulation was optimized by the self emulsifying efficiency of various combinations of oil and mix-surfactant evaluated by using pseudo-temary phase diagram. The preliminary stability of GB-SEMEDDS was evaluated by the variety of loading rate of GB and dispersed medium. The morphology, the particle size and the formulation stability were evaluated after diluting by 0.1 mol x L(-1) HCl.

RESULTThe blank self microemulsified system was composed of ethyl oleate-( cremophor EL-lecithin-ethanol, 4: 1:2) (40: 60), the loading dosage was 2.5%. Little influence of GB and emulsified medium was observed on the stability of GB-SEMDDS. After diluted with 0.1 mol x L(-1) HCl, the morphology of the microemulsion was homogeneous small spherical drops observed under electro-microscope. The particle size was (41.6 +/- 1.11) nm, the self microemulsifing time was around 2 min. The formulation was stable within 8 h, without significant changes in particle size and separation of drugs.

CONCLUSIONGB-SMEDDS is easy to prepare and its quality is stable. The solubility of GB was significantly improved by SMEDDS.

Chemistry, Pharmaceutical ; instrumentation ; methods ; Drug Delivery Systems ; instrumentation ; methods ; Emulsions ; chemistry ; Ginkgolides ; chemistry ; pharmacokinetics ; Lactones ; chemistry ; pharmacokinetics ; Particle Size ; Solubility ; Surface-Active Agents ; chemistry