S801 leukemic cell line established by our department was used as cells for interferon (IFN) induction and stimulating induction. S801 cell culture of concentration of 5-8x10~6 cells/ml was used for IFN induction and IFN titer of 3.55 Lg IU/ml was obtained by classical procedure. The IFN titer obtained S801 cell cultures after pretreatment with Ciwujia eleutherosides B. D and E or polysaccharide (PES) combined with priming procedure,then using Sendai Virus as an inducer was 10-20 times higher than that in cultures without any pretreatment. These Chinese medical herbs may be the ideal IFN helpinducers. This cell line may be applied for IFN production.

Candida shehatae xyl1 gene and Pichia stipitis xyl2 gene were amplified by PCR and the xyl1 and xyl2 were both placed under the promoter GAL of vector pYES2 to produce the recombinant expression vector pYES2-P12. Subsequently the pYES2-P12 vector was transformed into S. cerevisiae YS58 by LiAc to produce the recombinant yeast YSS8-12. It was indicate that the recombinant yeast YSS8-12 could converse xylose to ethanol with the xylose consumption rate of 81. 3%.

The crude toxin was extracted from hypha and culture solution of Phyllosticta commelimecola through three different polarity solvent: benzinum, puncificatum ethyl acetate and chloroform. The result indicated that the toxin secreted by Phyllosticta commelimecola not only was in hypha but also in culture solution and the extracting effect of ethyl acetate was the best. The soybean median and PSK media can be respectively used as solid and liquid culture media to produce toxin and grow mycelium. The optimal cultural conditions for producing toxin were temperature 32℃,cultured period 14d, cultured ways shaking of 150r/min.

In order to find a method suitable for purifying large amount of CD34(+) cells, from 5 cases who accepted autologous peripheral blood stem cell transplantation, CD34(+) cells were collected and enriched by using Isolex 300i (Nexell). Phenotypes were detected by flow cytometry and the biological viability were assayed by the colony-forming experiments and cell expansion experiment in vitro. The results showed that the number of mononuclear cells first collected was about (3.5 - 6.0) x 10(10) and (0.55 - 1.2)% of cells were CD34 positive. The number of positive production was about (2.0 - 3.0) x 10(8); the CD34(+) cells purity was (75 - 85)% and the yield was (40 - 65)%. The CD34(+) cells of positive production could expand up to 2 - 3 times when cultured with SCF + IL3 + FL + TPO + EPO in vitro. The results of colony-forming experiments demonstrated that the CD34(+) cells collected has enough colony-forming ability. All results showed the enriched CD34(+) cells with biological viability. In conclusion, the CD34(+) immunomagnetic isolation apparatus Isolex300i is suitable to clinical application for a large amount of CD34(+) cell enrichment.
Antigens, CD34 ; immunology ; Colony-Forming Units Assay ; Flow Cytometry ; Hematopoietic Stem Cells ; cytology ; immunology ; Humans ; Immunomagnetic Separation ; instrumentation ; methods ; standards ; Neoplasms ; blood

OBJECTIVESTo investigate whether antisense Bmi-1 plasmid could inhibit the proliferation of Jurkat cells.

METHODSThe antisense plasmid was constructed by PCR amplification of a 171 bp segment spanning Bmi-1 start codon and zinc finger structure and the PCR product was subsequently inserted reversely to plasmid pLNCX2. The final construct was confirmed through restriction enzyme digestion. G418 was added into the medium after the plasmid was successfully introduced into Jurkat cells by using lipofectin-mediated DNA transfection. The proliferation of Jurkat cells were determined by MTT and colony formation assays. Cell cycle was determined by flow cytometry. The p16 expression of Jurkat cells was studied by immunofluorescent histochemistry.

RESULTSThe growth rate of antisense Bmi-1 transfected Jurkat cells was significantly lower than that of the controls, and the colony forming capacity of the transfected cells decreased significantly (P < 0.01), the colony numbers being (90.7 +/- 9.07)/10(3) cells, (83.3 +/- 6.11)/10(3) cells and (56.0 +/- 5.56)/10(3) cells for control cells, empty plasmid transfected Jurkat cells and antisense Bmi-1 transfected Jurkat cells, respectively. The percentage of G, phase cells was increased and the p16 expression of antisense Bmi-1 transfected cells was significantly upregulated than that of control cells.

CONCLUSIONAntisense Bmi-1 can inhibit the growth and upregulate the expression of p16 of Jurkat cells in vitro.

Cell Cycle ; Cell Proliferation ; Gene Expression Regulation, Neoplastic ; Genetic Vectors ; Humans ; Jurkat Cells ; Nuclear Proteins ; genetics ; Oligonucleotides, Antisense ; genetics ; Plasmids ; genetics ; Polycomb Repressive Complex 1 ; Proto-Oncogene Proteins ; genetics ; Repressor Proteins ; genetics ; Transfection

To construct the Bac-to-Bac expression system of Bombyx mori nucleopolyhedrovirus (BmNPV), a transfer vector was constructed which contained an Escherichia coli (E. coli) mini-F replicon and a lacZ: attTN7: lacZ cassette within the upstream and downstream regions of the BmNPV polyhedrin gene. B. mori larvae were cotransfected with wild-type BmNPV genomic DNA and the transfer vector through subcutaneous injection to generate recombinant viruses by homologous recombination in vivo. The genomic DNA of budded viruses extracted from the hemolymph of the transfected larvae was used to transform E. coli DH10B. Recombinant bacmids were screened by kanamycin resistance, PCR and restriction enzyme (REN) digestion. One of the bacmid colonies, BmBacJS13, which had similar REN profiles to that of wild-type BmNPV, was selected for further research. To investigate the infectivity of BmBacJS13, the polyhedrin gene was introduced into the bacmid and the resultant recombinant (BmBacJS13-ph) was transfected to BmN cells. The budded viruses were collected from the supernatant of the transfected cells and used for infecting BmN cells. Growth curve analysis indicated that BmBacJS13-ph had a similar growth curve to that of wild-type BmNPV. Bio-assays indicated that BmBacJS13-ph was also infectious to B. mori larvae.

Objective:To observe the effect of Governor Vessel-unblocking and mind-refreshing acupuncture plus functional training on neural development in infants with brain damage and seek an effective method for early intervention of infantile brain damage.Methods:Eighty infants with brain injury were recruited and allocated to a treatment group and a control group by their visiting sequence,with 40 cases in each group.The control group received exercise training,40 min each session and 6 sessions a week,and tuina treatment,30 min each time and 6 times a week.Based on the treatment protocol for the control group,the treatment group additionally received Governor Vessel-unblocking and mind-refreshing acupuncture,3 times a week and 10 sessions as a course at a 2-week interval.Before the treatment and after 14-week treatment,the gross motor function measure (GMFM) and developmental quotient (DQ) of Bejing Gesell developmental scale were used to evaluate the development of the infants.Results:After the treatment,the GMFM score and DQs of Gesell scale all increased by different levels in the two groups,and the intra-group differences were statistically significant (all P＜0.05);the scores of the treatment group were superior to those of the control group,and the between-group differences were statistically significant (all P＜0.05).Conclusion:Governor Vessel-unblocking and mind-refreshing acupuncture plus functional training can significantly promote the development of gross motor and cognitive functions in infants with brain damage,and it is an early and effective intervention for infantile brain damage.

OBJECTIVETo investigate the effect of enhanced nutritional therapy on wound healing after endoscopic therapy in patients with liver cirrhosis and esophageal varices.

METHODSFifty patients with liver cirrhosis and esophageal varices were randomly divided into an enhanced nutritional therapy group (n = 25) and a control group (n = 25). The enhanced nutritional therapy group received one week of enhanced nutritional supplementation, including liver nutritional elements, prior to routine endoscopic therapy. The routine without any change to their diet. The rate of transformation and status of wound healing of esophageal varices were compared between the two groups.

RESULTSThe ratio of ulcers occurring at the injection site was lower in the enhanced nutrition group than in the control group (16/25 vs. 23/25; x2 = 5.711, P = 0.017). The enhanced nutrition group had only one case of minimal bleeding occurring during endoscopy as compared to the seven cases of bleeding in the control group (x2 = 5.357, P = 0.021). On average, the enhanced nutrition group required less sessions of endoscopic treatment to achieve eradication of esophageal varices than the control group (3.8 vs. 4.1; t = 2.069, P = 0.044).

CONCLUSIONPre-endoscopic enhanced nutritional therapy may benefit patients with liver cirrhosis and esophageal varices by promoting recovery of procedure-related local tissue injury and occlusion of varices.

Adult ; Endoscopy ; Esophageal and Gastric Varices ; etiology ; therapy ; Female ; Humans ; Liver Cirrhosis ; complications ; therapy ; Male ; Middle Aged ; Nutritional Support ; Wound Healing

OBJECTIVEBased on high fat diet induced the model of atherosclerosis (AS) in C57BL/6J mice, authors studied the effect of endurance exercise on the atherosclerostic formation.

METHODSForty eight 8-week-old C57BL/6J mice were divided into four groups randomly (n=12): control (group N), 12-week atherosclerosis model group(group H), 12-week atherosclerosis model plus 11-week treadmill training group (group H + E) and 22-week atherosclerosis model group (group HS). Then, we observed the effects of endurance exercise on the ultra structure of aorta by electron microscope.

RESULTSTwenty weeks of high fat diet could result in serious AS in mice while endurance exercise could significantly antagonize or restrain the occur of AS. In addition, 10 weeks of endurance exercise could alleviate the symptom of pathological changes which already happened on aorta wall.

CONCLUSIONIt indicated that endurance exercise could effectively prevent and cure AS that induced by high fat diet.

Animals ; Aorta ; ultrastructure ; Atherosclerosis ; etiology ; prevention & control ; Diet, High-Fat ; adverse effects ; Dietary Fats ; administration & dosage ; Male ; Mice ; Mice, Inbred C57BL ; Physical Conditioning, Animal ; physiology ; Physical Endurance ; physiology

Objective To develop the method of 16S rRNA gene clone library for tick bacterial flora analysis, and to analyze the detection effective of pathogens in tick and capacity of bacterial flora diversity. Methods Primers were designed according to the specific gene of Borrelia burgdorferi, Bartonella henselae, Anaplasma phagocytophilum, Ehrlichia chaffeensis and templates were choosen by positive PCR result to amplify the DNA extracted from the ticks. One set of primers targeting 16S rRNA gene conserved region were chosen to amplify certain fragments, DNA extraction, PCR reaction, cloning and sequencing. Nucleotide sequences were compared with GenBank database. Calculated Coverage values of clone library and Shannon-Wiener diversity index. Results Sixteen defined genus-or species-bacteria were detected in 103 valid sequences. Eight species were edge type (Clone No. > 5). Three kinds of pathogens were identified (Borrelia burgdorferi, Bartonella henselae and Rickettsia sp). Three kinds of pathogens were not edge type(Clone No. < 5). Coverage value was 96.11%, and Shannon-Wiener index was 2.40. Analysis results of cloning sequence showed that tick-parasitic bacteria mainly were α and γ deformation mycetes which accounted for 56.25% (9/16). Conclusions The 16S rRNA gene sequences technology could make relative quantitative of bacterial flora, and detect many kinds of pathogens in tick. It's a good method for detection of pathogens and bacterial flora analysis.