Nestin-positive progenitor cells isolated from human fetal pancreas have phenotypic markers identical to mesenchymal stem cells

AIM: To isolate nestin-positive progenitor cells from human fetal pancreas and to detect their surface markers and their capability of proliferation and differentiation into pancreatic islet endocrine cells in vitro.METHODS: Islet-like cell clusters (ICCs) were isolated from human fetal pancreas by using collagenase digestion. The free-floating ICCs were handpicked and cultured in a new dish. After the ICCs developed into monolayer epithelium-like cells, they were passaged and induced for differentiation. Reverse transcription polymerase chain reaction (RT-PCR), immunofluorescence stain, fluorescenceactivated cell sorting (FACS) and radioimmunoassay (RIA)were used to detect the expression of cell markers. RESULTS: (1) The monolayer epithelium-like cells had highly proliferative potential and could be passaged more than 16 timesin vitro; (2) RT-PCR analysis and immunofluorescence stain showed that these cells expressed both nestin and ABCG2, two of stem cellmarkers; (3) FACS analysis revealed that CD44, CD90and CD147 were positive, whereas CD34, CD38, CD45, CD71, CD117, CD133 and HLA-DR were negative on the nestin-positive cells; (4) RT-PCR analysis showed that the mRNA expression of insulin, glucagon and pancreaticduodenal homeobox gene-1 was detected, whereas the expression of nestin and neurogenin 3 disappeared in these cells treated with serum-free media supplemented with the cocktail of growth factors. Furthermore, the intracellular insulin content was detected by RIA after the induction culture.CONCLUSION: Nestin-positive cells isolated from human fetal pancreas possess the characteristics of pancreatic progenitor cells since they have highly proliferative potential and the capability of differentiation into insulinproducing cells in vitro. Interestingly, the nestin-positive pancreatic progenitor cells share many phenotypic markers with mesenchymal stem cells derived from bone marrow.

Mass screening of prostate cancer in a Chinese population:the relationship between pathological features of prostate cancer and serum prostate specific antigen

Aim:To investigate the pathological features of the prostate biopsy through mass screening for prostate cancer in a Chinese cohort and their association with serum prostate specific antigen (PSA).Methods:A total of 12 027 Chinese men in Changchun were screened for prostate cancer by means of the serum total prostate specific antigen (tPSA) test (by Elisa assay).Transrectal ultrasound-guided systematic six-sextant biopsies were performed on those whose serum tPSA value was >4.0 ng/mL and those who had obstructive symptoms (despite their tPSA value) and were subject to subse- quent pathological analysis with the aid of the statistic software SPSS 10.0 (SPSS.Inc.,Chicago.USA).Results:Of the 12 027 cases,158 (including 137 patients whose serum tPSA values were >4.0 ng/mL and 21 patients [serum tPSA <4. 0 ng/mL] who had obstructive symptoms) undertook prostate biopsy.Of the 158 biopsies,41 cases of prostatic carci- noma were found (25.9 %,41/158).The moderately differentiated carcinoma and poorly differentiated carcinoma ac- counted for 61% and 34 %,respectively.A significant linear positive correlation between the serum tPSA and the Gleason scores in the 41 cases of prostatic carcinoma (r=0.312,P<0.01) was established.A significant linear positive correlation between the serum tPSA value of the 41 prostatic carcinoma and the positive counts of carcinoma in sextant biopsies was established (r=0.406,P<0.01),indicating a significant linear relationship between serum tPSA and the size of tumor. Condusion:This study was the first to conduct mass screening for prostate cancer by testing for serum tPSA values and the first to investigate the pathological features of prostate cancer in a cohort of Chinese men.Our results reveal that the moderately differentiated carcinoma is the most common type of prostate cancer.This study also has shown that the serum tPSA value in prostate cancer is associated with the Gleason score and the size of tumor.

FINDING POTENTIAL LIGANDS FOR PDZ DOMAINS BY TAILFIT, A JAVA PROGRAM

Objective To deduce all potential ligands undiscovered experimentally by searching all the proteins containing same C-termini, which can bind a certain PDZ domain. Methods We developed a JAVA program for searching short exact sequence matches at C-terminus. According to the known C-termini, which PDZ domains recognized experimentally, Swissprot database has been searched by this program for all potential ligands. Results Some PDZ domains may have more potential ligand proteins, which are undiscovered yet experimentally. These bioinformatic results also provide clues for studying functions of hypothetical proteins and PDZ domains’ protein interactions in many different organisms. Conclusion The results may provide useful clues for discovering potential functions of hypothetical proteins and new functions of known proteins.

Solubility of disulfide-bonded proteins in the cytoplasm of Escherichia co/i and its “oxidizing” mutant

AIM: To study the influence of redox environment of Escherichia coli ( E. coli) cytoplasm on disulfide bond formation of recombinant proteins.METHODS: Bovine fibroblast growth factor (BbFGF) was selected as a model of simple proteins with a single disulfide bond and free cysteines. Anti-HBsAg single-chain Fv (HBscFv), an artificial multidomain protein, was selected as the model molecule of complex protein with 2 disulfide bonds. A BbFGF-producing plasmid, pJN-BbFGF,and a HBscFv producing-plasmid, pQE-HBscFv, were constructed and transformed into E. coli strains BL21(DE3)and M15[pREP4]respectively. At the same time, both plasmids were transformedinto a reductase-deficient host strain, E. coli Origami(DE3). The 4 recombinant E. coli strains were cultured and the target proteins were purified. Solubility and bioactivity of recombinant BbFGF and HBscFv produced in different host strains were analyzed and compared respectively.RESULTS: All recombinant E. colistrains could efficiently produce target proteins. The level of BbFGF in BL21(DE3)was 15-23% of the total protein, and was 5-10% in Origami (DE3). In addition, 65% of the BbFGF produced in BL21(DE3) formed into inclusion body in the cytoplasm,and all the target proteins became soluble in Origami (DE3). The bioactivity of BbFGF purified from Origami(DE3)was higher than its counterpart from BL21(DE3). The ED50of BbFGF from Origami(DE3) and BL21(DE3) was 1.6 μg/L and 2.2 μg/L, respectively. Both HBscFv formed into inclusion body in the cytoplasm of M15[pQE-HBscFv] or Origami[pQE-HBscFv]. But the supernatant of Origami[pQE-HBscFv] lysate displayed weak bioactivity and its counterpart from M15[pQE-HBscFv] did not display any bioactivity. The soluble HBscFv in Origami[pQE-HBscFv]was purified to be 1-2 mg/L and its affinity constant was determined to be 2.62×107 mol/L. The yield of native HBscFv refolded from indusion body in M15[pQE-H Fv] was30-35 mg/L and the affinity constant was 1.98×107 mol/L.There was no significant difference between the bioactivity of HBscFvs refolded from the inclusion bodies produced in different host strains.CONCLUSION: Modification of the redox environment of E. coli cytoplasm can significantly improve the folding of recombinant disulfide-bonded proteins produced in it.

Early diagnosis of prostate cancer using free/total prostate specific antigen ratio: a population based screening data

Aim: To evaluate the use of free/total prostate specific antigen ratio (fPSA/tPSA ratio) in improving the early diagnosis of prostate cancer. Methods: The fPSA/tPSA ratio in the serum was analyzed in 187 men with tPSA ranging between 4.0 and 20.0 μg/L. All of them underwent ultrasound guided sextant prostatic biopsy. The results were calculated by SPSS 10.0 software. Results: (1) When the tPSA was within the ranges of 4.0 - 10.0 and 10.0 -20.0 μg/L, the prostate cancer detection rate was 18.1 % and 22.5 %, respectively; (2) The area under the curve (AUC) was bigger in fPSA/tPSA than in tPSA (P<0.05) in all the men; (3) When the cut off value of fPSA/tPSA ratio was set at 0.25 and the tPSA at 4.0 - 10.0 μg/L and 10.0 - 20.0 μg/L, the diagnostic sensitivity of tPSA was 90.5 % and 87.5 %, respectively. Thus at the tPSA ranges of 4.0 - 10.0 and 10.0 - 20.0 μg/L, 26.7 % and 11.3 % of biopsies could be avoided, respectively. Conclusion: The use of fPSA/tPSA ratio can improve the prostate cancer detection rate and reduce unnecessary biopsies when tPSA is within the range of 4.0 - 20.0 μg/L.

Study on Perception of Potential Condom Use among Unmarried Migrants

Objective To understand the perception of potential condom use and its influencing factors so as to promote condom use among unmarried migrantsMethod A survey, using self-administered structured anonymous questionnaire, was conducted among 1 092 unmarried migrants aged 15～24 years old in urban area of Shanghai. Results The unmarried migrants had some knowledge of condom. However, their knowledge level and their attitude towards condom use were not so optimistic. Subjects' knowledge of contraceptives and STD/AIDS had a positive influence on their attitude toward condom use. Subjects with positive attitude to condom use were more likely to use condom in sexual activity (OR=1.61). Conclusion Unmarried migrants have lower rate of condom use if they have insufficient knowledge of condom. Therefore, it is essential to strengthen the sexual health education among unmarried sex-active migrants in order to encourage contraceptive use, especially condom use.

Dormancy Breaking and Storage Behavior of Garcinia cowa Roxb. （Guttiferae） Seeds： Implications for Ecological Function and Germplasm Conservation

Abstract: The dormancy breaking and storage behavior of Garcinia cowa Roxb. seeds were investigated. The seeds of G. cowa had 8–11 months dormancy in their natural habitat. Seeds were matured and dispersed at the end of the rainy season (mid-late August to late September) and were scatter-hoarded by rodents as food for winter after the seeds had fallen to the ground. Seedlings often emerged in the forest during the rainy season (May to August) the following year. Intact seeds of G. cowa failed to germinate after being sown at 30 °C for 120 d and the mean germination time (MGT) of seeds cultured in a shade (50% sunlight) nursery was 252 d. The most effective method of breaking dormancy was to remove the seed coat totally, which reduced the MGT to 13 d at 30 °C. Germination was also promoted by partial removal of the seed coat (excising the hilum and exposing the radicle) and chemical scarification (immersion in 1% H2O2 for 1 d). Unscarified seeds take up water rapidly in the first 96 h, but water was absorbed by the outside seed coat, without penetrating through it. The moisture content (MC) of G. cowa seeds was high (50% in fresh weight) at shedding. The seeds could tolerate desiccation to some extent, until the MC reached approximately 40%; below that, the viability decreases rapidly and all seeds died at approximately 17% of MC. Seed viability decreased rapidly when seeds were chilled at 4 °C; germination was 2% after storage for 1 week. Even stored at 10 °C, seeds began to be damaged after 4 weeks. Seed storage for 1 yr revealed that in both dry (relative humidity (35 ± 5)%) and moist (wet sand) storage conditions, seed viability declined, but germination percentages for seeds stored under moist conditions are better than for seed stored under dry conditions. Because of their low tolerance to desiccation, marked chilling sensitivity and relatively short lifespan, G. cowa seeds should be classified into the tropical recalcitrant category. The ecological implications of dormant recalcitrant seeds and cues on storing recalcitrant seeds were discussed.

A Quotient Space Approximation Model of Multiresolution Signal Analysis

In this paper, we present a quotient space approximation model of multiresolution signal analysis and discuss the properties and characteristics of the model. Then the comparison between wavelet transform and the quotient space approximation is made. First, when wavelet transform is viewed from the new quotient space approximation perspective, it may help us to gain an insight into the essence of multiresolution signal analysis. Second, from the similarity between wavelet and quotient space approximations, it is possible to transfer the rich wavelet techniques into the latter so that a new way for multiresolution analysis may be found.

MAIN FACTORS IN PREPARATION OFANTIBACTERIAL PARTICLES/PVC COMPOSITE

Zirconium phosphate containing silver was chosen as antibacterial particles in preparing antibacterial particles/PVC composite. The effect of surface property of the antibacterial particles and of their filler content on the properties of antibacterial particles/PVC composite was studied. The effect of the interfacial compatibility on mechanical properties of the composite was also discussed. Experimental results showed that the antibacterial PVC composite had good antibacterial property, reaching almost 100% bacteriostatic level at an antibacterial powder filler content of 1.5 phr.