1.Mutational Screening of Zuelacmycin-producing Strain RL-2
Li-Xia GOU ; De-Rong AN ; Shuang-Fa LIU ; Jing LI ;
Microbiology 2008;0(08):-
In this research, the zuelacmycin-producing strain Streptomyces venezuelaevar. qinlingensis RL-2 was used as original strain, the spore suspension of which was induced by UV, LiCl and the compound treatmeat (UV+LiCl) respectively. The zuelacmycin-high-yield strain UVL-108 was obtained by the treatment that the exposure time under UV irradition was 45 s and the concentration of LiCl was 0.4%. The heredity characters of mutant UVL-108 were stable in succesive six generations. The antibacterial activity and the fermentation titer of mutant UVL-108 were determinded by bidirectioned culture and mycelial linear growth respectively. The results demonstrated the antibaterial activity of mutant UVL-108 was improved by 77%, and the relative toxicity of fermentation to Phyricularia grisea was improved by two times compared with the original strain.
2.Screening and Identification of a New Elastase-producing Strain
Shuang-Fa LIU ; De-Rong AN ; Li-Xia GOU ; Na LI ;
Microbiology 2008;0(09):-
The study provide a theoretical basis for the industrial production of a elastase-high-yield strain which was isolated from the straw,and the selected strain was identified. The screening strategy included casein(skim milk) plate selecting and elastin(beef tendon) re-screening. And then,the morphological,physiological and biochemical characteristics as well as 16S rRNA sequence homology of the selected strain were studied. Finally,the strain LSF-97 which had a excellent decomposing ability to beef tendon was obtained. The results showed that the strain LSF-97 is relative to the Bacillus pumilus with 100 % similar in sequence under the phylogenetic tree,the morphological and physiological and biochemical characteristics are also consistent with pattern of bacteria. So it was identified as Bacillus pumilus.
3.Changs of B7 Family Costimulatory Molecules and Their Receptors Expression in Children with Asthma
ying, ZU ; cheng-rong, LI ; de-fa, LI ; yue-jie, ZHENG ; jing, LI
Journal of Applied Clinical Pediatrics 1994;0(04):-
Objective To explore the possible role of B7 family cell surface costimulatory molecules and their receptors in immunopathogenesis in children with asthma.Methods Forty children with acute exacerbation of asthma and 36 cases of age-matched healthy children were enrolled in the study.Interferon(IFN-?),IFN-? and lipopolysaccharide(LPS),phorbol 12-myristate 13-acetate and ionomycin or zero were used to stimulate peripheral blood mononuclear cells,flow cytometry was used to detect B7-1 and B7-2,B7H1,B7DC and B7H expression on CD14+ cells;CD4+ cells were separated by immune beads and stimulated by phorbol-12-myristate-13-acetate and ionomycin and fluorescence quantitative polymerase chain reaction was used to detect CD28,cytotoxic lymphocyte associated antigen-4(CTLA-4),programmed death-1(PD-1)and inducible costimulator(ICOS) mRNA expression.Results 1.The levels of B7-1 and B7-2 expression in children with asthma were higher than those of healthy control group [B7-1:(6.68?3.97)% vs(1.74?0.69)%;B7-2:(10.87?5.99)% vs(1.58?0.75)% Pa
4.Relationship between Imbalance of Suppressors of Cytokine Signaling 1/Suppressors of Cytokine Signaling 3 and Abnormal Activation of Monocytes/Macrophages in Children with Asthma
ying, ZU ; cheng-rong, LI ; de-fa, LI ; yao-jie, ZHENG ; hong-ling, MA
Journal of Applied Clinical Pediatrics 2006;0(16):-
Objective To investigate the relationship between the imbalance of suppressors of cytokine signaling 1(SOCS1)/SOCS3 and abnormal activation of monocytes/ macrophages in children with acute-phase asthma, and explore the molecular mechanism of chronic inflammatory process on airway.Methods The present study enrolled 20 asthmatic children and 20 age-matched normal children. Dual-color flow cytometric analysis was performal to detect the percentage of CD_ 80 、CD_ 86 expressing on CD_ 14 ~+ cell. Reverse transcriptase-polymerase chain reaction and real-time PCR were used to analyze SOCS1,SOCS3 expression in monocytes/ macrophages.Results The proportions of CD_ 80 ,CD_ 86 expressed on CD_ 14 ~+ cell in children with asthma were significantly higher than those in control subjects(CD_ 80 :7.0% vs 1.70%),(CD_ 86 :11.37% vs 2.03%),all P
5.Relationship between combined multigene detection and response to chemotherapy and prognosis in epithelial ovarian carcinomas
Li LI ; Xiu-Ying MO ; Wei ZHANG ; Min-Yi WEI ; Fa-Long CHEN ; De-Sheng YAO ;
Chinese Journal of Obstetrics and Gynecology 2000;0(09):-
Objective To evaluate the relationship between combined muhigene detection and response to chemotherapy and prognosis in epithelial ovarian carcinomas(EOCS).Methods A total of 80 ovarian tissue samples taken from the surgical specimens of patients with EOCS of our hospital in the last two decades who had received chemotherapy "after surgery were paraffin-embedded.The samples were divided into 2 groups,good prognosis group(patients who survived more than 2 years,n=46)and poor prognosis group(patients who survived less than 2 years,n=34).The expression levels of ToPo-Ⅱ,Ki-67,MGMT, PCNA,p27,p53,pl6,P-gp,LRP,GST-?,bcl-2,C-myc,Fas,bax,MSH2,MRP and BCRP were investigated by the combination of tissue arrays and immunohistoehemical streptavidin-biotin peroxidase(SP) method in all samples.Data were analysed with SPSS 12.0 for windows.Results There were statistically significant differences in the positive expression levels of P-gp,BCRP,MGMT,MSH2,p27 and p16(62%, 50% and 50% in poor prognosis group vs 33%,28% and 28% respectively,P
6.Expression of synaptotagmin I and effect of thyroxine treatment in the prefrontal cortex of adult hypothyroid rats
Xi, WANG ; Hui-yun, YANG ; Jian-yue, WANG ; Li, GUI ; De-fa, ZHU
Chinese Journal of Endemiology 2010;29(3):239-242
Objective To observe the expression of synaptotagmin I(syt I)protein in the prefrontal cortex of adult-onset hypothyroidism rats and the effects of replicated therapy in different doses of thyroid hormone on the syt I protein.Methods All 44 aduh male Sprague-Dawley rats were divided into 4 groups randomly according to their body mass:hypothyroidism group,routine dosage thyroxine treatment group,high dosage thyroxine treatment group and control group.The adult male Sprague-Dawley rats were replicated to the adult-onset hypothyroidism and treatment models with propyhhiouracil(PTU).The levels of serum T3,T4 were assayed by the radioimmunoassay method and the level of the syt I protein in the molecular layer,external granular layer,external pyramidal layer,internal granular layer and internal pyramidal layer in prefrontal cortex was analyzed by immunohistochemistry.Results In the hypothyroidism group,the levels of serum T3 and T4[(0.34±0.04),(43.01±2.95)nmol/L]were significantly lower than those in the control group[(0.65±0.15), (55.20±3.56)nmol/L, F value: 6.026,5.940,4.503,P<0.05 or <0.01 ], the levels of the syt I protein in the molecular layer(0.018±0.010), external granular layer (0.020±0.007), external pyramidal layer(0.013±0.008), internal granular layer(0.011±0.005), internal pyramidal layer(0.024±0.013) of prefrontal lobe were significantly lower compared to the control group[(0.028±0.010,0.031 ± 0.010,0.028 ± 0.010,0.022 ± 0.008,0.038 ± 0.013), F value: 5.697,8.965,14.668,13.597,6.807,P<0.05 or <0.01 ]. In the routine dosage of the thyroxine treatment group, the levels of serum T3,T4 [(0.63 ±0.05), (55.04 ± 3.77)nmol/L] were not significantly different compared to the control group(F value: 3.162,0.367,all P>0.05), and the level of the syt I protein in the molecular layer, external granular layer, external pyramidal layer, internal granular layer and internal pyramidal layer in prefrontal cortex showed a significant improvement of the syt I protein(0.027 ± 0.013,0.025 ± 0.009,0.022 ± 0.008,0.020 ± 0.010,0.033 ± 0.010), which were similar to that of the control group(F value: 0.094,2.208,2.467,0.350,0.693, all P>0.05). In the high dosage thyroxine thyroid hormone treatment group, the levels of serum T3 and T4[ (1.11 ± 0.10), (96.68 ± 6.42)nmoL/L] were higher than the control group(F value: 6.291,12.031, all P<0.01), the expression of the syt I protein(0.028 ± 0.008,0.031 ±0.011,0.026 ± 0.012,0.023 ± 0.011,0.038 ± 0.010) were not significantly different compare to the control group (F value: 0.001,0.019,0.111,0.061,0.001, all P>0.05). Conclusions The expression of the syt I protein in the prefrontal cortex of adult-onset hypothyroidism can be decreased, which can be reversed by routine dosage of thyroxine treatment.
7.Observation and contrast of anaerobe culturing result using three kinds of device
Chang-Fa SHAO ; De-Ren LI ; Yun-Long YANG ; Ken-Ying JIANG ;
Chinese Medical Equipment Journal 1993;0(05):-
The positive ratio is 74 percent using the plate anaerobe culturing device made by author, higher than using common anaerobic box and anaerobic jar.
8.Construction of a novel Schistosoma japonicum DNA vaccine pBK-Sj14-3-3 and studies on its immunoprotection in mice.
De-fa LI ; Yue-sheng CHEN ; Ying ZU ; Ji-long SHEN
Chinese Journal of Preventive Medicine 2004;38(3):193-195
OBJECTIVETo prepare Sj14-3-3 DNA vaccine and observe its immunoprotection against Schistosoma japonicum in mice.
METHODSThe Sj14-3-3 gene was amplified by reverse transcription-polymerase chain reaction (RT-PCR) and subcloned into eukaryotic expression vector pBK. The recombinant plasmid pBK-Sj14-3-3 was extracted, purified and inoculated into BALB/c mice by intramuscular injection. Mice were attacked by Schistosoma japonicum cercariae and then killed. Adult worm and egg were counted, respectively. Diameter of the egg granulomas in the liver of infected mice was measured.
RESULTSElectrophoresis on 1% agarose gel showed that the product of RT-PCR and the inserted fragment of recombinant plasmid digested with EcoR I and Xho I had the same size, about 765 bp, confirming the latter was the 14-3-3 encoding gene by nucleotide sequencing. Adult worm load declined by 27%, average egg load of per gram (EPG) of the liver tissues by 79%, average egg production per couple of adult worm (EPWP) by 51%, and mean diameter of egg granulomas by 29% in vaccinated mice.
CONCLUSIONThe recombinant plasmid pBK-Sj14-3-3 was successfully constructed, which had some immunoprotection against Schistosoma japonicum in infected mice, indicating its potential to be vaccine candidate molecule of Schistosoma japonicum.
14-3-3 Proteins ; genetics ; immunology ; Animals ; Antibodies, Helminth ; blood ; Antigens, Helminth ; genetics ; immunology ; Cloning, Molecular ; DNA, Helminth ; genetics ; Female ; Helminth Proteins ; genetics ; immunology ; Membrane Proteins ; genetics ; immunology ; Mice ; Mice, Inbred BALB C ; Parasite Egg Count ; Rabbits ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; Schistosoma japonicum ; genetics ; immunology ; Schistosomiasis japonica ; immunology ; prevention & control ; Vaccines, DNA ; immunology
9.Roles of CD8+ CD28- T regulatory cells in acute infectious mononucleosis in children.
Ying ZU ; Cheng-rong LI ; Zu-xiang MA ; De-fa LI ; Xiao-ling FU
Chinese Journal of Pediatrics 2007;45(3):208-211
OBJECTIVETo explore the role of CD(8)(+)CD(28)(-) T regulatory cells (Tr) in the immunological pathogenesis of acute infection with Epstein-Barr virus in children.
METHODSThe present study enrolled 25 children with infectious mononucleosis (IM) and 25 age-matched healthy children. Flow cytometric analysis was performed to detect the percentage of CD(3)(+), CD(3)(+)CD(4)(+), CD(3)(+)CD(8)(+), CD(8)(+)CD(28)(+) by determining the ratio of positive cells in lymphocytes. Reverse transcriptase-polymerase chain reaction (RT-PCR) and real-time PCR were used to analyze IL-6, IL-10, IFN-gamma expression in CD(8)(+)CD(28)(-) Tr cells and ILT-3, ILT-4 expression in monocytes/macrophages.
RESULTSThe proportions of CD(8)(+)CD(28)(-)T cells in children with acute-phase IM was significantly higher than those in the controls (P < 0.01). The expression level of IL-6, IL-10, IFN-gamma, ILT-3, ILT-4 mRNA significantly increased compared to those of the controls (P < 0.01).
CONCLUSIONThe CD(28) expressed on CD(8)(+) T cells in vivo is gradually lost with age and CD(8)(+)CD(28)(-) cells increase up 50% to adult. EBV can directly infect B cells, trigger CD(8)(+) CTL response and destroy the target cells to cause serious immunopathological lesion. Therefore we speculate that the expansion of CD(8)(+)CD(28)(-) Tr cells in children with IM may be an adaptive immune response to avoid serious inflammation and autoimmune reactions.
CD28 Antigens ; immunology ; CD8-Positive T-Lymphocytes ; immunology ; Case-Control Studies ; Child ; Child, Preschool ; Cytokines ; immunology ; Epstein-Barr Virus Infections ; immunology ; Female ; Flow Cytometry ; Herpesvirus 4, Human ; Humans ; Infectious Mononucleosis ; immunology ; virology ; Male ; Membrane Glycoproteins ; metabolism ; Receptors, Cell Surface ; metabolism ; Receptors, Immunologic ; metabolism ; T-Lymphocytes, Regulatory ; immunology
10.Treatment outcome of repeat gamma knife radiosurgery for primary trigeminal neuralgia
Chun-Fa DU ; Zhi-Yuan ZHANG ; De-Sheng XU ; Li-Gao ZHENG
Chinese Journal of Neuromedicine 2008;7(10):1044-1047
Objective To assess the feasibility, indications and therapeutic effect of repeatgamma knife radiosurgery for treatment of primary trigeminai neuralgia. Methods From January 1995to February 2006, 277 patients with primary trigeminal neuralgia underwent stereotactic radiosurgerytargeting the cistemal trigeminal nerve with a maximal central dose of 70-80 Gy. A second gamma kniferadiosurgery with a maximal central dose of 65-80 Gy was performed in 23 patients with refractory and15 with recurrent trigeminal neuralgia after the primary surgery. The 50% isodose line encompassed thetarget area for radiation. The trigeminal root entry zone was included in a single target area in 33 patients,and in 5 patients, two target areas were chosen. Results All patients were followed up for 12 to 108months (mean 55 months) by telephone. The pain relief lasted for 1 to 180 days (mean 120 days) after thesecond gamma knife radiosurgery. The primary stereotactic radiosurgery resulted in a complete or partialpain relief rate of 91.7%(254/277). In the 23 patients with refractory trigeminal neuralgia, the secondradiosurgery achieved a pain relief rate of 73.9% (17/23), significantly lower than the rate of 100%(15/15) in the 15 patients with recurrent trigeminal neuralgia (P=0.045). Facial numbness or paresthesiaoccurred in 30 patients (10.8%) after the primary radiosurgery, and in another 8 patients (21.1%) after thesecond surgery. Conclusion Gamma knife stereotactic radiosurgery is safe and effective and causesfew complications for relieving trigeminal neuralgia. A repeat gamma knife stereotactic radiosurgery isalso effective for management of recurrent or refractory trigeminal neuralgia, especially in recurrentcases.