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Objective To observe the influence of IL-12,IL-15 on CIK cell in the normal culture;to observe the anti-tumor effect in the circumstance of different combination of cytokines,and to provide a new insight for preparing high effective and qualified CIK cell in vitro.Methods The optimal concentrations of IL-2,IL-12 and IL-15 were determined,respectively.After the peripheral blood from healthy blood donors was collected,monocytes were selected and co-cultured with different cytokines into different groups,as group A(IL-2 normal culture group),group B(IL-2 and IL-12 group),group C(IL-2 and IL-15 group),group D(IL-2,IL-12 and IL-15 group),and group E(cytokine control group).The monocytes in different groups were calculated by globulimeter,the activity of cells was detected by Trypan blue staining,positive ratio of CD3,CD8,CD56 on the celluar membrane was detected by flow cytometry,and the anti-tumor effect of CIK to SMMC-7721 was detected by MTSmethod,inthedayof0,5,10,15,20 after the culture.Results Statistical analysis indicated that,the proliferation multiplication of CIK cells was significantly higher in group B,group C and group D after 10,15 and 20 days of culture than those in group A(P＜0.05);and group D had higher proliferation multi-plication than that of group C(P＜0.05).The percentage of CD3 + CD8 +,CD3 + CD56+ in CIK cell membrane in group B,C,D was significantly higher than that in group A after 15 and 20 days of culture (P＜0.05).The percentage of CD3+ CD8+,CD3+ CD56+ in CIK cell membrane in group D was significantly higher than that in group B after 15 and 20 days of culture (P＜0.05).The killing rate of CIK cells for liver cancer in each group at 10,15,20 days of culture was significantly higher than that of group A when the target target ratio was 5 ∶ 1 (P＜0.05).The killing rate of CIK cells for liver cancer in group D,C at 10,15,20 days of culture was significantly higher than that of group B(P＜0.05).Conclusion IL-12and IL-15 could improve the proliferation of CIK cells,and IL-15 also has the effect of enhancing CIK cells the tumor-killing to SMMC-7721 activity.
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Objective To observe the correlation between protein C activity-dependent clotting time-normalized ratio (PCAT-NR) and the related blood coagulation parameters,e.g.,fibrinogen (Fib),factor Ⅶ coagulant activity (Ⅶ:C),factor Ⅷcoagulant activity (F Ⅷ:C),antithrombin (AT),D-dimer (DD) in patients with acute cerebral infarction.Methods One hundred cases of patients who were diagnosed as acute cerebral infarction according to clinical manifestations and imaging examinations were taken as the test group and 75 healthy subjects were taken as control group.The values of Fib,FⅦ:C,FⅧ:C,AT,PCAT-NR,DD were tested and the difference between the two groups were compared.The differences of Fib,FⅦ:C,FⅧ:C,DD and AT between declined PCAT-NR group and normal PCAT-NR group in the patients with acute cerebral infarction were analyzed.The correlations of PCAT-NR with other coagulation parameters in acute cerebral infarction cases were compared.Results The values of Fib (3.38 ± 1.25) g/L,F Ⅶ:C (130.5 ± 15.9) ％,FⅧ:C (135.8 ± 43.1) ％ and DD (2.12:±:3.01) mg/L in the acute cerebral infarction group were significantly higher than those of control group,while the values of AT (83.94 ± 14.95) ％ and PCAT-NR (0.87 ± 0.23) in test group were significantly lower than those the control group (P＜0.05).The values of Fib (4.03 ± 1.25)g/L,FⅦ:C (138.2 ±6.9)％ and FⅧ:C (151.5 ± 54.9)％ of PCAT-NR declined group in the patients with acute cerebral infarction were significantly higher than those of PCAT-NR normal group (P ＜ 0.05),while the values of DD,AT were not statistically different between two groups (P ＞ 0.05).The values of PCAT-NR were significantly negatively correlated with Fib,FⅧ:C and DD in the patients with acute cerebral infarction (r =-0.484,-0.356 and-0.473,respectively (all P ＜ 0.05).There was no correlation of PCAT-NR with FⅦ:C and AT (P ＞ 0.05).Conclusion The PCAT-NR decline was associated with high coagulation state in patients with acute cerebral infarction.This decline has some correlation with high level of blood clotting factor Ⅷ and Fib.
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ObjectiveTo evaluate “main line teaching-topic design” based classroom discussion method in immunology teaching. MethodsStudents of five-year class of pharmacology of Grade 2008 and Grade 2009 were selected to sit the innovating teaching.The teaching methods included main line teaching,topic design,classroom discussion and experimental operation.The evaluation of the effect was analyzed by the way of a questionnaire and comparing test scores.ResultsQuestionnaire survey results show that more than 73.5％ of experimental class students thought that the “main line teaching-topic design” based classroom discussion method helps to stimulate their learning enthusiasm and improve comprehensive ability.By T test,the difference of the average scores of experimental class and control class students was statistically significant ( P=0.0028 ).ConclusionThe “main line teaching-topic design”based classroom discussion method is accepted as an effective approach of immunology teaching and worth to extensive application.
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OBJECTIVE To investigate the resistant mechanisms of Candida albicans to azoles at molecular level.METHODS NCCLS M-27 protocols were used to test the in vitro susceptibilities of 102 C.albicans strains isolated from the patients with recurrent vulvovaginal candidiasis(RVVC) against fluconazole(FLC) and itraconazole(ITC) to screen the FLC-and ITC-resistant C.albicans isolates;six pairs of primers,A1-A2,B1-B2,C1-C2,D1-D2,E1-E2 and F1-F2 were respectively to amplify gene CYP51 of 4 strains with FLC-and ITC-resistance.The PCR products were sequenced and analyzed to identify the mutation sites by compared with the sequence of gene CYP51 of referenced C.albicans strain in NCBI site of Internet.RESULTS The analysis of full length sequence of CYP51 from 4 FLC-and ITC-resistant strains showed that from total 32 mutation sites there were 4 significant site mutations,where the mutation of GAT to GAC at 116 caused the substitution of D by E(E266D in two strains);GCC to GGT at 117 caused the substitution of A by G(A117G in 1 strain);GAA to GAC at 266 caused the substitution of E by D(E266D in 2 strains);and GTT to ATT at 488 caused the substitution of I by V(V488I in 1 strain).The site mutations of 266 and 488 were tested in 1 strain of 4 strains.CONCLUSIONS The CYP51 total gene of 4 strains has been checked out.Of FLC and ITC-resistant C.albicans alignment in this time,find out 4 significant bp mutations.Causing its amino acide change,among them,A117G has not be interrelated report still now.The details of mechanism need to be further studied.
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Objectives To explore the effects of Baicalin and Berberine against Trichomonas vaginalis(TV) in vitro. Methods The micro-double-dillution assay is used to measure the Minimal lethal concentration (MLC) of Baicalin and Berberine against 6 freshly isolated strains of TV, using metronidazole as control. Results MLC of Baicalin and Berberine against 6 isolated strains in vitro of TV ranges from 0.125 to 0.5 mg/mL and 0.25 to 1.0 mg/mL, respectively. The means of MLC are 0.229 mg/mL and 0.417mg/mL, respetively, with statistically significant difference (t=2.666, P
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Aime To study the anti-C. albicans activity of baicablin,and to explore its anti-C.albicans mechanism preliminarily. Method ① The minimum inhibitory concentration (MIC) and the minimum fungicidal concentration(MFC) of baicalin against C.albicans were determined by microdilution method. ② By means of incorporation experiment of isotope-labelled precursor, the incorporation inhibitation rates of [ 3H]-TdR,[ 3H]-UdR,[ 3H]-leucine were measured after treatment with various concentrations of baicalin respectively. Result ① The MIC and MFC of baicalin (above purity 98%) against C albicans(1?107 cfu?L -1) were both 1.0 g?L -1. The MIC and MFC of baicalin (above 98% purity) against C albicans(2.5?107 cfu?L -1) both 2.0 g?L -1. ② The dose of baicalin of 0.5, 1 and 2 MIC could obviously inhibit incorporation of the precursors and there were significant differences among the inhibition rate of three precursors incorporation (P0.05). Conclusion ① Baicalin had strong anti-C albicans activity in vitro. ② Baicalin(above purity 98%) could obviously inhibit incorporation of [ 3H]-TdR, [ 3H]-UdR and [ 3H]-leucine into C albicans and the incorporation inhibition rate of [ 3H]-UdR was highest among them, suggesting that the anti-C albicans mechanism of baicalin may be involved with the inhibition of DNA, RNA and protein synthesis.