The yeast fusant ZFF-28, which is high in biomass production and rich in selenium, was constructed after mutagenesis and protoplasts fusion between yeast strains. The total selenium content of ZFF-28 is 1.8 and 1.0 times higher than that of the parental strains Saccharomyces cerevisiae ZY-67 and Saccharomyces kluyveri SZY-198 respectively. Using single factor tests and a L16(4(3) x 2(1)) orthogonal design, the cultivation conditions was optimized as: 50mL culture in 250mL shake flasks in molasses containing 6% sugar and 60microg/mL Se at 28 degree C for 25h at 220 r/min, with the initial pH adjusted to 6.0 - 6.5. Under the optimized conditions, the biomass (dry weight) reached 8.2g/L and the Se content of the cells reached 2050microg/g, with organic and inorganic Se contents being 91% and 9% respectively.
Biomass ; Hydrogen-Ion Concentration ; Saccharomyces ; genetics ; growth & development ; metabolism ; Saccharomyces cerevisiae ; genetics ; metabolism ; Selenium ; metabolism ; Selenium Compounds ; metabolism

Adult ; Bone Neoplasms ; diagnostic imaging ; therapy ; Combined Modality Therapy ; Curettage ; methods ; Embolization, Therapeutic ; methods ; Female ; Follow-Up Studies ; Giant Cell Tumor of Bone ; diagnostic imaging ; therapy ; Humans ; Male ; Sacrum ; Time ; Tomography, X-Ray Computed ; Treatment Outcome ; Young Adult

Objective To investigate the influence factors of birth defects. Methods The congenital malformational fetuses born from 13 week of gestation to 7 days after birth were selected as the study group between April 1st, 2009 and March 31st, 2010. The health born fetuses were set as control in the same period. Case-control and the three-level of monitor network of birth defects were used in the study in the participating 75 hospitals (Qingdao Women and Children's Medical Center, Affiliated Hospital of Medical College Qingdao University, Qingdao Municipal Hospital, etc.) . The study and control group's parents were interviewed by an uniformed questionnaire which was designed specially with influence factors of birth defects. Results (1) There are 466 congenital malformational fetuses in the total of 77 231 fetuses collected in 75 hospitals. The congenital malformational rate accounts for about 6. 034‰. The top six defect diseases were congenital heart disease (112 cases) , total harelip (cleft lip; cleft lip with palate; 85 cases) ,polydactyly (53 cases) , neural tube defects (38 cases) , congenital hydrocephalus (37 cases) and limb reduction defect (27 cases) in turn, which amounts to 353 cases (54.48% , 353/648). (2) Their mother education level in the birth-defect group (25. 6%) were significantly lower than that in control group (30. 0% , P < 0. 05). (3) The rate of passive smoking, drinking, raising pets of the parents in birth-defect group were significantly higher than that in control group (P < 0. 05). (4) The rate of exposure to harmful chemical and physical factors of mothers in birth defects group (13.9% and 20. 5% , respectively) was higher than that in control group (1.1% and 11.7% , respectively) , the difference between which were significant (P < 0. 01) . The rate of disease (34. 3%) , fever (13. 1%) , taking drugs (33. 8%) in pregnancy period in birth defect group were higher than that in control group (13. 5% , 1.5% and 9. 9% ,respectively) , the difference between which were significant (P <0. 01). The rate of bad moral irritation to the mother in pregnancy in birth defect group (11. 3%) was significantly higher than that in control group (1.4% , P<0.01). (5) There were 19 cases (2.9% , 19/648) with family heredity medical history in birth defect group, but there were none in the control group, the difference between which were significant (P<0. 01). There were 89 cases (13.7% , 89/648) with unusual birth history of their mothers in birth defect group, but there were 31 cases (4. 8% , 31/650) in control group, the difference between which were significant (P<0. 01). (6) Conditional Logistic Regression model was used for univalent and multivariate analysis. The results showed that main influence factors were identified as having important effect on birth defects, including mothers' exposure to harmful chemical factors (OR=13. 46) , disease (OR=3. 37) ,taking drugs (OR=2. 20) , exposure to bad moral irritation (OR=5. 44) , food-choosy (OR=1. 90) ,anemia (OR=1. 52) in gestational period, polyembryony (OR=4. 40) , father drinking (OR=1. 55) .While it was protective factors to supplement microelements such as the calcium iron and nutrient, etc. in pregnancy period (OR=0. 45). Conclusions First, the main birth defects were congenital heart disease, total harelip (cleft lip; cleft lip with palate) , polydactyly, neural tube defects, congenital hydrocephalus and limb reduction defect in turn. Second, the main influence factors identified as having important effect on birth defects were mothers' exposure to harmful chemical factors, ill, taking drugs, exposure to bad moral irritation, food-choosy, anemia in gestational period, polyembryony, father drinking. But it is protective factors to supplement microelements such as the calcium iron and nutrient, etc. in pregnancy period. Finally, it is the important part to prevent the birth defects by reducing and controlling dangerous factors in pregnancy period.

Seventy-two strains of endophytic fungi were isolated from the healthy bark, branches and leaves of Cephalotaxus hainanensis L.. Sixty-eight of them were morphologically classified into Fungi Imperfecti, thirty-three sporulated were identified to five genera. For those did not sporulate, one was identified to Rhizoctonia sp., the rest were tentatively classified into Mycelia Sterilia. Four were identified to Basidiomycetes. The result indicated the endophytic fungi of C. hainanensis show a degree of tissue specificity. There were significant differences about the quantity, genera and composition between the fungi isolated from bark and those from branches and leaves.

The rice gall dwarf disease, caused by the Rice gall dwarf virus (RGDV) is a serious disease occurring in rice in many regions of Guangdong province. As a basis to control the disease we have studied the genomic diversity of a variety of isolates from different locations. Genome segment 8(S8), encoding a main outer capsid protein (Pns8) of RGDV five isolates (BL, CH, DQ, GZ, XY) from Guangdong province was cloned and sequenced. The results revealed that all the S8 segments of the five isolates consisted of 1 578 nucleotides and had a single open reading frame (ORF) extending for 1 301 nucleotides from nucleotide 21 which encoded a polypeptide of 426 amino acids with an estimated molecular weight of 47.4 kDa. The S8 full-length sequence and the ORF sequence shared 97.3%-98.8% and 97.3%-99.1% nucleotide sequence identities within the five Chinese isolates, and shared 94.8%-95.6% and 95.0%-96.0% identities with those of the Thailand isolate respectively. The deduced amino acid sequence of Pns8 in GZ isolate was identical to that in the Thailand isolate, while the amino acid sequence variability of Pns8 within five Chinese isolates ranged from 0.5% to 2.1%. These results indicate that the S8 segment of RGDV is highly conserved in different isolates from different locations. The S8 cDNA from the XY isolate was cloned into the plasmid vector pET-28b(+) and a fused expression protein with an apparent molecular mass of 51kDa was specifically detected in an analysis of Escherichia coli Rossetta(DE3)Ⅱcells. To our knowledge, this is the first report on analysis of the RGDV segment 8 sequence and genetic comparison of different RGDV isolates and their protein expression.

Objective To observe the effect of bilateral movement training on upper extremities dysfunction in stroke patients in convalescent phase. Methods 52 patients with hemiplegia were randomly divided into treatment group (n=26) and control group (n=26). The treatment group accepted bilateral upper extremities movement training, and the control group accepted routine neurodevelopment training mainly with affected upper extremities, for 6 weeks. They were assessed with Fugl-Meyer Assessment upper-extremity section (FMA-UE) and modified Barthel Index (MBI) before and after treatment. Results The FMA-UE and MBI scores improved in both groups after treatment (P<0.01), and improved more in the treatment group than in the control group (P<0.001). Conclusion Bilateral movement training may improve upper extremity function and activities of daily living more effectively for stroke patients in convalescent phase.

The effects of different physical and chemical factors on hairy root growth and flavonoids production were studied in suspension culture of Saussurea medusa hairy root in 1/2 MS medium. The results showed that the following culture conditions, nitrogen concentratiaon (involved NH4+ and NO3-), 30 mmol/L; the ratio of ammonium to nitrate, 5:25; the combination of 2% sucrose and 3% glucose; 0.5 mg/L GA3; 0.5 mg/L IBA; initial pH 5.8; light cycle, 18 h/d (3500lx); temperature, 24 degrees C; shaker revolutions per minute, 100 r/min, were favourable to hairy root growth and flavonoids production. Under the above culture conditions, up to 12.8 g/L (DW) of hairy root and 1922 mg/L of flavonoids were obtained after 21 days of culture. The content of total flavonoids in hairy root was 15%, which was about 25 times as that in the wild plantlet.
Culture Media ; Flavonoids ; biosynthesis ; Plant Roots ; growth & development ; metabolism ; Saussurea ; growth & development ; metabolism ; Temperature ; Tissue Culture Techniques

Hairy root clones of Saussurea involucrata transformed with Agrobacterium rhizogenes strains R1601, R1000, and LBA9402 were established to investigate the flavonoid production. Opine synthesis and PCR analysis confirmed the integration of the T-DNA fragment of Ri plasmid from A. rhizogenes strain R1601 into the transformed root genome. The frequency of hairy root formation from root segments, which were pre-cultured 2 days in N6 solid medium without plant growth regulators, amounted to 100% following infection with R1601 strain of A. rhizogenes. The transformed roots were kept in hormone-free N6 liquid medium in the dark at 25 degrees C, 110r/min and routinely subcultured every 20 - 24 days. One hairy root clone, which grew vigorously with lateral branches, was periodically examined for the ability to produce flavonoid. The maximum of biomass and flavonoid yield achieved 66.7 g/L (fresh weight) and 102.3mg/g dry weight after incubation 20 days. The calli were induced from the hairy root culture in the presence of 0.5mg/L IBA and intact plantlets were regenerated from these calli. The regeneration plantlets from hairy roots, in which the flavonoid content were 53% in that of untransformed plants, weren't different in growth and morphology of the untransformed plantlets. Therefore plant regeneration from hairy roots may be also a means for producing transformed S. involucrata plants. Hairy root cultures of S. involucrata clearly showed higher flavonoid contents compared to the wild plant or the regeneration seedlings. As the wild S. involucrata grows only in special regions with peculiar climate, and cultivation of this species in a normal climate has been unsuccessful so far. The success in obtaining a method for high production of flavonoid might very well be one of the solutions for this problem in the future.
Culture Techniques ; Flavonoids ; biosynthesis ; Plant Roots ; growth & development ; Rhizobium ; physiology ; Saussurea ; growth & development

Administration, Oral ; Adult ; Alanine Transaminase ; blood ; Antiviral Agents ; administration & dosage ; therapeutic use ; DNA, Viral ; blood ; Female ; Guanine ; administration & dosage ; analogs & derivatives ; pharmacology ; therapeutic use ; Hepatitis B e Antigens ; blood ; Hepatitis B virus ; drug effects ; Hepatitis B, Chronic ; drug therapy ; immunology ; Humans ; Lamivudine ; administration & dosage ; pharmacology ; therapeutic use ; Male ; Treatment Outcome ; Young Adult

OBJECTIVETo investigate the effects of shikonin on the proliferation, expression of CXCR4 and the migratory responses to CXCL12 in colorectal carcinoma cell line SW480.

METHODSThe proliferation of SW480 cells was assessed by MTT assay. Cell surface expression of CXCR4 was determined by flow cytometry. The migratory ability was determined by Transwell.

RESULTSShikonin inhibited the proliferation of SW480 cells in time- and concentration-dependent manner. The expression rate of CXCR4 in SW480 cells was 99.1%. After application of shikonin 0.01 micromol/L, 0.1 micromol/L and 1.0 micromol/L for 24 h, the expression rate of CXCR4 decreased to 76.0%, 59.1% and 35.5% respectively (F=1098.041, P <0.001), and the CXCL12-induced SW480 cell migratory inhibition rate was 25.2%, 38.5% and 55.7% respectively (F=48.970, P <0.001).

CONCLUSIONBesides having inhibiting tumor cell proliferation effect, Shikonin may also play a role in anti-metastasis via down-regulating the expression of CXCR4 and reducing the CXCL12-induced migratory response in colorectal carcinoma cell.

Cell Line, Tumor ; Cell Proliferation ; drug effects ; Chemokine CXCL12 ; metabolism ; Down-Regulation ; Humans ; Naphthoquinones ; pharmacology ; Receptors, CXCR4 ; metabolism