Animals ; Cell Line, Tumor ; Female ; Humans ; Mammary Neoplasms, Experimental ; diagnosis ; metabolism ; pathology ; Mice ; Mice, Nude ; Neoplasm Transplantation ; Organometallic Compounds ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Radioisotopes ; Radiopharmaceuticals ; pharmacology ; Random Allocation ; Rhenium ; Tumor Burden ; drug effects ; bcl-2-Associated X Protein ; metabolism

Objective To detect the expression of CD133 in hepatic cell lines SMMC7721 and bcl-7402, and to investigate the possibility of CD133 as the surface marker of liver cancer stem cells. Methods The cell cycle and expression of CD133 in hepatic cell lines SMMC7721 and bcl-7402 were detected by flow eytometry. Magnetic cell sorting was used to isolate CD133-positive and CD133-negative cells. The differences in morphology, proliferation and differentiation between CD133-positive and CD133-negative cells were observed and analyzed by one-way ANOVA and u test. Results The percentages of CD133-positive cells in SMMC7721 and bcl-7402 cell lines were 0.7% -1.0% and 1.7% -8.9%, respectively. The percentages of CD133-positive cells in G0>/G1> phase in the 2 cell lines were 85.3% and 89.4%, which were significantly higher than CD133-negative cells and unsorted cells (F = 14.49, 38.84, P <0.05). The in vitro proliferation capability of CD133-positive cells was greater than that of CD133-negative cells and unsorted cells, especially during day 1-3 and day 5-7 (F =49.32,784.04, 89.91, 152.83, P < 0. 05). During the cultivation, the proportion of the CD133-positive cells decreased as time passed by, and the proportion of CD133-positive cells was nearly the same as unsorted cells on day 15 (u =O. 271, P <0.05). Conclusions CD133-positive ceils have strong capability of proliferation and differentiation in SMMC7721 and bcl-7402 cell lines in vitro. CD133 is one of the surface markers of liver cancer stem cells.

Objective Aliginic sodium diester(ASD) liposome was prepared and its aggregation stability was evaluated.Methods ASD liposome was made by reverse phase evaporating method from egg yolk lecithin,compared with blank liposome,the size distributions and mean particle diameters of ASD liposome and blank liposome were measured respectively before and after 40 hours heating under 37℃.Results Under the condition of n(PC)∶n(CHOL) = 1∶1,the mean size diameter of ASD liposome was 4.24?m.The mean size diameter of ASD liposome changed only 0.26?m after cultivation,however,the value of blank liposome was 1.35?m.Conclusion ASD loaded in the liposome enhanced the physical stability of the liposome.

Objective To evaluate the stability and biodistribution of a novel SPECT-MRI bi-functional agem 99Tcm-Gd-DTPA-DG in tumor-bearing nude mice. Methods DTPA-DG was synthesized and then conjugated with Gd2O3 to generate Gd-DTPA-DG. The tumor-bearing nude mice were scanned by MRI to evaluate the tumor targeting ability of Gd-DTPA-DG. The orthogonal experiment was applied to optimize pH value of reaction medium and reaction temperature. The radiolabeling efficiency was measured by thin layer chromatography. The distribution of 99Tcm-Gd-DTPA-DG in nude mice was evaluated by scintigrapy in vivo. The % ID/g was measured at different time after intravenous injection of 99Tcm-Gd-DTPA-DG. Results The tumor was significantly enhanced by Gd-DTPA-DG with MRI. The radiochemical purity of 99Tcm-Gd-DTPADG was about 98.5% and remained 96.2% at room temperature for 6 h. The tumor was well visualized by 99TcmGd-DTPA-DG SPECT at 2 h after injection. The tumor uptake was (1.48 ±0.12) %ID/g, and the rumor to muscle radioactivity ratio was 2.91. Conclusions MRI contrast of Gd-DTPA-DG may enhance tumor detection. 99Tcm-labeled Gd-DTPA-DG may be useful for tumor imaging and might have a potential role as a SPECT-MRI bi-functional agent.

The purpose of this study is to explore the feasibility of wheat germ agglutinin (WGA) modified liposome as a vehicle for ophthalmic administration. Liposome loaded with 5-carboxyfluorescein (FAM) was prepared by lipid film hydration method. WGA was thiolated and then conjugated to the surface of the liposome via polyethylene glycol linker to constitute the WGA-modified and FAM-loaded liposome (WGA-LS/FAM). The amount of thiol groups on each WGA molecule was determined, and the bioactivity of WGA was estimated after it was modified to the surface of liposome. The physical and chemical features of the WGA-modified liposome were characterized and the ocular bioadhesive performance was evaluated in rats. The result showed that each thiolated WGA molecule was conjugated with 1.32 thiol groups. WGA-LS/FAM had a mean size of (97.40 +/- 1.39) nm, with a polydispersity index of 0.23 +/- 0.01. The entrapment efficacy of FAM was about (2.95 +/- 0.21)%, and only 4% of FAM leaked out of the liposome in 24 h. Erythrocyte agglutination test indicated that after modification WGA preserved the binding activity to glycoprotein. The in vivo ocular elimination of WGA-LS/FAM fitted first-order kinetics, and the elimination rate was significantly slower than that of the unmodified liposome, demonstrating WGA-modified liposome is bioadhesive and suitable for ophthalmic administration.
Absorption, Physicochemical ; Adhesiveness ; Administration, Ophthalmic ; Animals ; Drug Carriers ; Eye ; metabolism ; Fluoresceins ; chemistry ; Liposomes ; administration & dosage ; chemistry ; pharmacokinetics ; Male ; Particle Size ; Polyethylene Glycols ; chemistry ; Rats ; Rats, Sprague-Dawley ; Wheat Germ Agglutinins ; administration & dosage ; chemistry ; pharmacokinetics

AIMTo investigate the distribution of c-myb, an oncoprotein, in mouse oocytes-cumulus cell complex and sperm immunohistochemically.

METHODSTo study the effect of c-myb on mouse fertilization in vitro, various concentration of c-myb antisense-oligodeoxynucleotides (c-myb ASODNs) were incubated with sperms and oocytes during fertilization. To explore the possible mechanism involved in fertilization, the relationship between c-myb ASODNs and GABA or dbcAMP or Verapamil or Progesterone in fertilization was also observed by immunohistochemical methods.

RESULTSc-myb oncoprotein was observed on the nucleus of cumulus cell and head of sperm. c-myb ASODNs inhibited the rate of fertilization in vitro in a dose-dependent way. The fertilization rates of the control group, low (5 micromol/L), medium (10 micromol/L), high (20 micromol/L) concentration c-myb ASODNs groups and nonsense tat oligodeoxynucleotides (20 micromol/L) group were 34.97%, 30.89%, 20.14%, 16.68%, 34.47%, respectively. All of GABA, Progesterone and dbcAMP inversed the c-myb ASODNs inhibition effects on fertilization rate, but neither of them showed significant effect on the percentages of immunohistochemical stain of Myb on sperm and cumulus cells. By contrast, Verapamil inhibited the fertilization rate. Co-treated with c-myb ASODNs, Verapamil showed synergic inhibiting effects on the fertilization with c-myb ASODNs. Verapamil also inhibited the expression of Myb on head of sperm. The fertilization rates of the control group, medium (10 micromol/L) concentration c-myb ASODNs group, GABA group, P4 group, Verapamil group, dbcAMP group were 34.81%, 22.96%, 40.83%, 39.12%, 7.46%, 40.61%, respectively.

CONCLUSIONc-myb ASODNs is closely correlated with fertilization. Verapamil can inhibit fertilization in vitro through regulating Myb expression of sperm, while GABA, dbcAMP and Verapamil may affect the process of fertilization through the way other than Myb expression.

Animals ; Bucladesine ; pharmacology ; Female ; Fertilization ; physiology ; Fertilization in Vitro ; Male ; Mice ; Mice, Inbred Strains ; Oligodeoxyribonucleotides, Antisense ; pharmacology ; Oocytes ; physiology ; Proto-Oncogene Proteins c-myb ; metabolism ; Spermatozoa ; physiology ; Verapamil ; pharmacology ; gamma-Aminobutyric Acid ; pharmacology

Objective To elucidate the biocompatibility differences of 4 dialyzers with different membranes in maintenance hemodialysis (MHD)patients. Methods A total of 60 MHD patients were enrolled in the prospective,randomized,control,cohort study.In baseline,synthetic polysulfone(PS)membrane dialyzer was used in all the patients for at least 3 months.Then the patients were randomly divided into three groups:ployethersulfone(PES)membrane group,cellulose triacetate (CTA)membrane group,and synthetic polymethylmethacrylate(PMMA)membrane group.Study duration was 6 months.No dialyzer was reused.The biocompatibility markers were detected repeatedly at different time points and compared with each other in different dialyzer groups. Results The blood levels of high sensitive C reactive protein,interleukin-1β and interleukin-13 were not significantly different among different groups on every time point.However,the blood complements levels and WBC count were significantly different among four kinds of dialyzer.When the dialyzers changed from PS to PMMA membrane,C3a levels and WBC count changed significantly(P＜0.05).Moreover,the change of C5a level was significantly different between PES group and PMMA group on month 3(P＜0.05). Conclusion There are some differences of biocompatibiliy among different dialyzer membranes.

Adult ; Aged ; Apoptosis ; Female ; Humans ; Laryngeal Neoplasms ; metabolism ; pathology ; Male ; Middle Aged ; Neoplasm Staging ; Receptors, Tumor Necrosis Factor, Member 6b ; metabolism

Objective:To assess the efficacy of acupuncture plus Methadone in treating heroin withdrawal syndrome.Methods:According to the requirements of evidence-based medicine, the inclusion criteria, exclusion criteria and retrieval strategy were set for original documents. Randomized controlled trials (RCTs) involving acupuncture plus Methadone in treating heroin withdrawal syndrome were retrieved from China National Knowledge Infrastructure Database (CNKI), Wanfang Academic Journal Full-text Database (Wanfang), Chinese Biomedical Literature Database (CBM), Chongqing VIP Database (CQVIP), PubMed, Cochrane Library and EMBASE. According to the Cochrane Handbook for Systematic Reviews of Interventions, each included trial was assessed strictly including risk of bias and quality evaluation. Meta-analysis and descriptive-analysis were conducted using the RevMan 5.3 software.Results:A total of 8 trials involving 931 patients were included. Meta-analysis showed that there was no statistical significance [RR=1.05; 95%CI (0.99, 1.11);P=0.11] in comparing total effective rate between the acupuncture plus Methadone and Methadone alone; the cure rate of acupuncture plus Methadone was higher than that of Methadone alone [RR=1.45; 95%CI (1.19, 1.78);P=0.0003]; the relapse rate of the acupuncture plus Methadone was lower [RR=0.691; 95%CI (0.60, 0.80);P<0.00001]. On the basis of these results, the method recommended by the Grading of Recommendations Assessment, Development and Evaluation (GRADE) was taken to evaluate the quality of the evidence. It showed that the level of the evidence was low and it was weakly recommended.Conclusion:According to the included trials, acupuncture plus Methadone in treating heroin withdrawal syndrome is more effective and better than Methadone alone, but it still needs more high-quality, large sample, and polycentric researches to confirm it.

Hespintor is an unknown function protein that was got from hepatoblastoma cell lines HepG2 by suppression subtractive hybridization technique (SSH), sequence analysis showed that the protein is a new member of secretory type of Kazal type serine protease inhibitor (Serpin) family, and has high homology with esophageal cancer related gene 2 (ECRG2). The coding sequence of Hespintor's Kazal domain was subcloned into prokaryotic expression vector pET-40b(+), then transformed into Rosetta (DE3). A recombinant protein about 42 kDa in the form of inclusion body was optimization expressed by inducing with 0.25 mmol/L IPTG, 30 degrees C for 5 h. and its specificity was confirmed via Western blotting. The recombinant protein was purified by metal chelate affinity chromatography (MCAC) and anion-exchange chromatography. The preliminary experimental result showed that the recombinant protein can inhibit trysin hydrolysis activity specifically. The result clearly demonstrated that Hespintor, as a novel member of Serpin, would be valuable in developing anti-tumor agents.
Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; Hep G2 Cells ; Humans ; Proteinase Inhibitory Proteins, Secretory ; Recombinant Proteins ; biosynthesis ; genetics ; Serine Peptidase Inhibitors, Kazal Type ; Serine Proteinase Inhibitors ; biosynthesis ; classification ; genetics