1.Construction of mouse peritonitis models of Enterococcus
Journal of Chongqing Medical University 1987;0(01):-
Objective:To construct mouse peritonitis models of Enterococcus for the study of the putative virulence factor of Enterococcus.Methods:We used the standard strain OG1RF and sprE gene deficient mutant of Enterococcus faecalis to construct mouse peritonitis models.And the index of peritonitis was detected to measure the virulence decrease of sprE mutant. Results:It was showed that 50%lethal dose(LD50)of sprE mutant was 7 times than that of standard strain OG1RF,and its survival rate was also prominently that of OG1RF in the mouse peritonitis model;while the change of blood leukocytes of sprE mutant group was lower than that of standard stain at 6 hour and 12 hour;So were the TNF-? level in peritoneal fluid and active TNF-? in peritonitis at 6 hour.Conclusion:The peritonitis models of Enterococcus was constructed successfully,which would be used to study other putative virulence factor of Enterococcus face-ali.
2.Establishment of rabbit endocarditis models caused by enterococcus
Journal of Chongqing Medical University 1986;0(04):-
objective:To establish Rabbit endocarditis models caused by Enterococcus faecalis for studying virulence of Enterococcus.Methods:Standard strain OG1RF and sprE gene deficient mutant of Enterococcus faecalis were used to establish Rabbit endocarditis models.The index of endocarditis was detected to show virulence change.Results:The size,weight and Colony count of vegetation of sprE mutant groups were smaller than those of wild-type OG1RF groups in a rabbit endocarditis model.Conclusion:The Rabbit endocarditis models caused by Enterococcus were establised successfully,which would be used to study other virulence factor of Enterococcus faecalis.
3.Curcumin derivative C085 inhibits proliferation of K562/G01 cells and activity of Bcr-Abl kinase in vitro
Ying WU ; Lixian WU ; Jianhua XU
Chinese Pharmacological Bulletin 2016;32(7):1004-1011
Aim To find new kinase inhibitors that o-vercome the imatinib resistance in the treatment of chronic myeloid leukemia ( CML ) by synthesizing C085, a novel derivative of curcumin , and testing its activities against wild-type( WT) and imatinib-resistant mutant Abl kinases , as well as in imatinib-resistant CML cells in vitro.Methods Cell proliferation and apoptosis were examined using MTT assay and flow cy-tometry, respectively;Kinase activity was measured u-sing Kinase-Glo Luminescent Kinase Assay Platform in recombinant WT and mutant ( Q252H, Y253F, and T315I) Abl kinases.The phosphorylation levels of Bcr-Abl initiated signaling proteins were analyzed using Western blotting .Colony forming units ( CFU ) growth was used to test the effects of C 085 on human leukemia progenitor/stem cells.Results C085 suppressed the growth of imatinib-resistant K562/G01 cells and po-tently inhibited both WT and mutant ( Q252H, Y253F, and T315I) Abl kinase activities in a non-ATP com-petitive manner with the values of IC 50 at low nanomole levels.C085 dose-dependently down-regulated Bcr-Abl kinase activity in K562/G01 cells as judged by auto-phosphorylation and Stat 5 , Crkl phosphorylation , and inhibited the phosphorylation of downstream targets Raf,Mek and Erk with protein content reducing .C085 could directly impact mitochondrial PT hole and make it open, which prevents the activation of caspase cas-cade reaction and induces the apoptosis .Furthermore, C085 significantly suppressed CFU growth , implicating that C085 could inhibit human leukemia progenitor/stem cells.Conclusion C085 may inhibit K562/G01 cells through inhibiting Bcr-Abl kinase activity and down-regulating the downstream signal proteins .Di-rectly acting on mitochondrial PT hole and then activa-ting apoptosis-associated proteins are also involved in the pro-apoptotic effect of C085.C085 is a promising compound for the treatment of CML patients with Bcr-Abl kinase domain mutations that confer imatinib re-sistance .
4.Establishment of mouse septic peritonitis model with low pathogenic bacteria
Tao LUO ; Wenxiang HUANG ; Lixian WU
Journal of Third Military Medical University 1984;0(01):-
0.05). Conclusion The application of SRFE can decrease MLD effectively when form mice septic peritonitis models, especially for those bacteria with low pathogenicity.
5.Development and application of a perioral force measurement system for infants with cleft lip and palate
Yaqi ZHENG ; Lixian ZHANG ; Guofeng WU
Journal of Practical Stomatology 2016;32(4):490-494
Objective:To develop a perioral force measurement system for the infants with cleft lip and palate.Methods:The peri-oral force measurement system of infant with cleft lip and palate is composed of hardware and software.The sensor is metal cantilever. The measurement ranges are 0 -20 and 0 -1 00 g/cm2 ,and the precision is 0.1 g/cm2 .The system was used in 4 cases of infants with unilateral cleft lip and palate before and after cheiloplasty.The results were analyzed by SPSS 1 9.0 software.Results:Before cheilo-plasty the perioral force of labial frenum area was (1 .79 ±0.94)g/cm2 ,that of angulus oris area of normal side and cleft side was (5. 41 ±1 .01 )g/cm2 and (3.1 2 ±1 .55)g/cm2 (P <0.05);after cheiloplasty:the perioral force of labial frenum area was (1 2.73 ±3. 51 )g/cm2 ,that of angulus oris area of normal side and cleft side was (7.64 ±1 .64)g/cm2 and (7.27 ±1 .89)g/cm2 .Conclusion:The perioral force measurement system can be used to measure the perioral force of the infants with cleft lip and palate.
6.MLVA based Mycobacterium tuberculosis clinical isolates genotyping analysis f rom HIV co-infection patients in Dali area,China
Heng NIE ; Xudong WANG ; Lixian WU
Chinese Journal of Zoonoses 2015;(10):923-926
A new method based on the multiple locus variable number tandem repeat analysis (MLVA) was applied for the genotyping of combined HIV Mycobacterium tuberculosis in Dali to investigate the genotyping and distribution pattern of Myco‐bacterium tuberculosis clinical isolates with MLVA .Mycobacterium tuberculosis clinical isolates were selected from Dali area , and the polymorphism of VNTR locus was tested with PCR .The clustering of genotype was analyzed by BioNumerics (6 .6) . Result showed that 15 VNTR loci of 61 combined HIV Mycobacterium tuberculosis clinical isolates were analyzed respectively . There were obvious polymorphisms of VNTRs .The discrimination power of these loci appeared different from each other ,with the biggest Hunter‐Gaston index (0 .839) loci was MIRU26 ,and the smallest one (0 .341) loci was MIRU4 .The clustering of genotype showed that these strains could be categorized into 5 gene clusters and 61 genotype ,the proportions of cluster Ⅰ was the biggest one ,51 .6% were cluster Ⅰ which including 32 strains .The standard strain H37Rv was belongs to cluster Ⅱ .Its indicated that there are obvious polymorphisms of VNTRs of combined HIV Mycobacterium tuberculosis clinical isolates in Da‐li .The main genotype was Beijing family genotype .
7.Effects of curcumin derivatives C085 on K562 cells and its mechanism
Ying WU ; Ruijia CHEN ; Lixian WU ; Jianhua XU
Chinese Pharmacological Bulletin 2015;(6):870-875
Aim To explore the anti-proliferation and apoptotic effects of C085, a curcumin derivative, on K562 cells and its mechanism. Methods MTT assay and flow cytometry were used to examine cell prolifera-tion and apoptosis, respectively. The phosphorylation levels of Bcr-Abl initiated signaling proteins were ana-lyzed using Western blot. Results The results showed that C085 suppressed the growth of K562 cells and the IC50 value was about 5-fold lower than that of Cur. C085 also induced significant apoptosis on K562 cells in 24 hours when compared with imatinib. Western blot results demonstrated that C085 down-regulated the phosphorylation of Bcr-Abl in K562 cells in a dose-de-pendent manner. The phosphorylation of Stat 5 and
Crkl, which were downstream signaling proteins of Bcr-Abl kinase, was also inhibited by C085. C085 caused the opening of mitochondrial PT holes as detected by JC-1 fluorescent, which inhibited Bcl-2 and enhanced Bax , then induced apoptosis. Conclusion C085 in-hibited BCR-ABL+ K562 cells through inhibiting BCR-ABL kinase activity and down-regulating its down-stream signal proteins. Directly acting on mitochondrial PT hole and then activating apoptosis- associated pro-teins are also involved in the pro-apoptotic effect of C085 .
8.Intervention and intervention of cisplatin combined with gemcitabine or vinorelbine in the treatment of patients with non-small cell lung cancer
Julian WU ; Li SUN ; Lixian WU ; Yan LI ; Qiaoling DING
Chinese Journal of Biochemical Pharmaceutics 2017;37(9):319-321
Objective To investigate the effect of psychological intervention on patients with non-small cell lung cancer treated with cisplatin or gemcitabine or Changchun vinorelbine. Methods 64 cases of patients with non-small cell lung cancer after surgery, nursing for patients in the group given basic treatment, routine care control group, the observation group based on routine nursing and psychological nursing effects were compared between the two groups. Results No significant difference of anxiety and depression in the patients of the two groups before nursing, after grouping nursing, the observation group improved significantly; compared two groups of patients with quality of life score, visible two group before the intervention had no significant difference after intervention group was clearly observed after the patients in the observation group were higher than that of the control group, comparison between groups were indicates that the difference is obvious (P<0.05). Conclusion Chemotherapy in non-small cell lung cancer after surgery for patients with psychological intervention, compared to conventional nursing, can improve the psychological status of patients with better, improve the quality of life of patients, so it is worthy of reference in clinical use.
9.Prevalence of virulence genes in Streptococcus pneumoniae strains isolated from clinical patients
Yishan DONG ; Wenxiang HUANG ; Tao LUO ; Cheng ZHANG ; Lixian WU
Chinese Journal of Microbiology and Immunology 2009;29(2):177-180
Objective To investigate the prevalence of virulence genes(ply, pspA, nanA, lytA, psaA) among Streptococcus pneumoniae recently isolated from clinical patients. Methods The 133 strains were isolated from patients in three teaching hospitals in Chongqing from 2006 to 2008. Polymerase chain reaction was used to screen for virulence genes (ply, pspA, nanA, lytA, psaA). Results The positive rate of lytA, psaA, ply, nanA and pspA in 133 clinical isolates were 94.7%, 85.0%, 82.7%, 84. 2% and 60.2%, respectively. The positive rates of the lytA, psaA, ply, nanA and pspA genes in 87 common serotypes isolates was 100%, 87.4%, 86.2%, 89.7%, 67.8%, respectively. Conclusion The total positive rates of five virulence genes in the 133 clinical strains were high. The positive rates of five genes in the com-mon serotypes isolates were higher than those in the no-common serotypes. These genes are important virulence genes of Streptococcus pneumoniae. They could be candidates for protein vaccine of Streptococcus pneumoniae.
10.The therapeutic effects of survivin antisense nucleic acid combined with paclitaxel on subcutaneous xenograft mouse model of Balb/c
Lixian WU ; Lisen HUANG ; Ruijia CHEN ; Jue TIAN ; Fang KE
Journal of Xi'an Jiaotong University(Medical Sciences) 2015;(4):467-471
Objective To explore the therapeutic effects of combined application of survivin antisense nucleic acid and taxol in subcutaneous xenograft mouse model of Balb/c and to preliminarily investigate the mechanism of the anticancer effects.Methods The model of subcutaneous tumor was established by hypodermic injection of C26 cells into Bal b/c mice.The mice were then randomly divided into five groups through the internal tumor injection:the blank group (C),lipo2000 group (L),paclitaxel group (T),survivin antisense nucleic acid group (A),and survivin antisense nucleic acid combined with paclitaxel group (A+T).We observed tumor growth,determined cell apoptosis by TUNEL method,and detected the expression of survivin by Western blot.Results ① All treatment groups had T/C<60%,which was significantly different from that of group L (P <0.05);the intervention was proved effective in vivo .The tumor inhibition rate of mice tumor weight showed that there were significantly curative effects in groups T,A and A+ T compared with that in group C (P < 0.05 ).The antitumor activity of paclitaxel (tumor inhibition rate of 21.82%±0.84%)could be improved by more than 59% through combination therapy (tumor inhibition rate of 54.1 6% ± 0.32%)concerning inhibition of tumor weight growth.② TUNEL method detected apoptotic cells:The tumor cells hardly had apoptosis in the blank group while T group and A group had a certain number of apoptotic cells.The experiment results suggested that PTX could promote tumor cell apoptosis,and that not only A+T strengthened the effect in killing tumor cells,but also the synergy of both could influence tumor resistance and ultimately make the effect in promoting tumor cell apoptosis conspicuous.③ The expression of survivin protein:The results showed that the expression of survivin protein in group A + T was obviously decreased without the expression of β-actin affected;it did not change significantly in group C compared with group L.The ratio of the A-value in groups T,A and A+T was 0.895 ±0.01 1,0.704 ±0.121 and 0.345 ± 0.01 9,respectively.Analysis of variance t-test showed that the expression level in group A+T obviously differed from that in groups C,L,A and T (P <0.05).Conclusion The combined therapy of survivin antisense nucleic acid and taxol can promote tumor cell apoptosis by downregulating the expression of survivin protein,reduce the body’s resistance to drugs and create synergetic effects.