Objective: The purpose of this study is to investigate the dynamic changes in the numbers and functions of CD8~+T cells and NK cells in patients with advanced ovarian cancer undergoing first line chemotherapy,so as to identify whether there was a "window period" of anti-tumor immune suppression reverse after chemotherapy.Methods: Peripheral blood samples from each ovarian cancer patient were obtained before (S_0) and at day 5-7 (S_1),day 12-14 (S_2) and day 25-28 (S_3) after Chemotherapy in 13 patients.The numbers and proportions of CD3~+,CD4~+,CD8~+ and nature killer (NK) cells were analyzed by flow cytometry technique.The percentages of specific IFN-γ-secreting CD8~+ cells were also calculated after that peripheral lymphocytes had been stimulated with self tumor lysates.Cytotoxicity of NK cells against K562 cells was detected by LDH releasing assay.Results: The numbers of CD3~+,CD4~+,CD8~+T cells and NK cells reduced to the lowest on S1.Compared to those of the control group,and the percentages of IFN-γ-secreting CD8~+T cells were remarkably higher on S_1,S_2 and S_3 when CD8~+T cells were stimulated with autologous tumor antigen,and the percentage of CD8~+IFN-γ~+ cell reached the highest on S2.No significant differences of NK cell cytotoxicity against K562 cells were found on S_1,S_2 and S_3 compared to S0.Conclusion: Paclitaxel and carboplatin induce lymphopenia,which triggers the temporary immune reconstitution.During immune reconstitution the enhanced priming of CD8~+T cell response by autologons tumor antigen is found while the function of NK cells does not change significantly.It probably turns out that the" window period"during immune reconstitution offers a best opportunity for cancer immunotherapy.

Objective:The purpose of this study is to investigate the dynamic changes in the numbers and functions of CD8+T cells and NK cells in patients with advanced ovarian cancer undergoing first line chemotherapy,so as to identify whether there was a "window period" of anti-tumor immune suppression reverse after chemotherapy.Methods:Peripheral blood samples from each ovarian cancer patient were obtained before (S0) and at day 5-7 (S1),day 12-14 (S2) and day 25-28 (S3) after chemotherapy in 13 patients.The numbers and proportions of CD3+,CD4+,CD8+ and nature killer (NK) cells were analyzed by flow cytometry technique.The percentages of specific IFN-?-secreting CD8+cells were also calculated after that peripheral lymphocytes had been stimulated with self tumor lysates.Cytotoxicity of NK cells against K562 cells was detected by LDH releasing assay.Results:The numbers of CD3+,CD4+,CD8+T cells and NK cells reduced to the lowest on S1.Compared to those of the control group,and the percentages of IFN-?-secreting CD8+ T cells were remarkably higher on S1,S2 and S3 when CD8+ T cells were stimulated with autologous tumor antigen,and the percentage of CD8+ IFN-?+ cell reached the highest on S2.No significant differences of NK cell cytotoxicity against K562 cells were found on S1,S2 and S3 compared to S0.Conclusion:Paclitaxel and carboplatin induce lymphopenia,which triggers the temporary immune reconstitution.During immune reconstitution the enhanced priming of CD8+T cell response by autologous tumor antigen is found while the function of NK cells does not change significantly.It probably turns out that the "window period" during immune reconstitution offers a best opportunity for cancer immunotherapy.

Objective To investigate the changes of Tc1/Tc2 profiles and the cytotoxic function of CD8~+ cells in ovarian cancer patients undergoing paclitaxel and carboplatin chemotherapy, so as to identify whether there is a "window period" of anti-tumor immune suppression reverse after chemotherapy. Methods Blood samples from each ovarian cancer patient were obtained before (S_0) and at day 5-7 (S_1), day 12-14 (S_2) and day 25-28 (S_3) after chemotherapy in 13 patients. Thirteen age-matched healthy female volunteers were enrolled as a control group. Flow cytometry technique was employed to analyze the proportion of Tc1/Tc2. The numbers of specific IFN-γ, secreting CD8~+ cells were also calculated after peripheral lymphocytes had been stimulated with self tumor lysates. Results The proportion of Tel in CD8~+ cells increased re-markably on S2 while the proportion of Tc2 in CD8~+ cells remained no significant changes after chemothera-py. The ratio of Tc1 to Tc2 cells reached the highest on S_2. IFN-γ/secreting CD8~+ T cells also increased re-markably on S_2, especially when CD8~+ T cells were stimulated with autologous tumor antigen. Conclusion Paclitaxel and carboplatin induce the changes of Tc1/Tc2 profile and augment anti-tumor immune response by immune reconstitution. It probably turns out that the "window period" during immune reconstitution offers a best opportunity for cancer immunotherapy.

Objective To compared the condition of DSA and MRA for detection the lesion vessels in elder patients with recurrent ischemic cerebrovascular disease(ICVD).Methods 52 patients with ICVD,including 41 cases of cerebral infarction(CI) and 11 cases of transient ischemic attack(TIA) were detected by DSA,and the 44 cases of them were detected by MRA too.Results The vascular anomalies were detected in 43 cases(82.69%) of the 52 patients by DSA.Among them,39 cases(75%) had vascular stenosis,in which 31 cases(75.61%) were CI,and the other 8 cases(72.73%) were TIA.The aneurysm were founfd out in 2 cases,and vascular dysplasia and variation were also found in each 1 case by DSA.The vascular stenosis were found in 10 cases(22.73%) of 44 patients by MRA,in which 8 cases(18.18%) were CI,while 2 cases(4.55%) were TIA.The detection rate of lesion vessels in DSA was significantly higher than that in MRA(P

AIM: We investigated the numbers and proportions of lymphocyte subsets in advanced ovarian cancer patients undergoing chemotherapy, so as to identify whether there is immune reconstitution after chemotherapy, and seek rational chemo-immunotherapy strategies in ovarian cancer treatment. METHODS: Blood samples from each ovarian cancer patient were obtained before (S_0) and at day 5-7 (S_1), day 12-14 (S_2) and day 25-28 (S_3) after chemotherapy in 13 patients. Flow cytometry technique was employed to analyse the numbers, proportions of CD3~+, CD4~+, CD8~+, CD4~+CD25~+ Treg and memory-like phenotype lymphocyte subsets. RESULTS: Lymphopenia was observed at S1 after chemotherapy, but lymphocytes were found gradually recovered after S1. The numbers of CD3~+, CD4~+, CD8~+T cells and CD4~+CD25~+ Treg cells reduced to the lowest on S_1. The proportions of CD8~+ T cells and CD4~+CD25~+ Treg cells reduced to the lowest on S_1 and S_2 respectively. The proportions of CD45RO~+ memory T cells increased significantly on S2 while the proportion of CD8~+CD45RA~+ T cells decreased remarkably on S_2. CONCLUSION: Paclitaxel and carboplatin induce lymphopenia at day 5-7 after chemotherapy, then comes the temporary immune reconstitution. It probably turns out that the window period during immune reconstitution offers a best opportunity for cancer immunotherapy.

Objective Vaccination is a most effective method for the prevention of severe diseases caused by pandemic influenza and microRNA ( miRNA) mediated gene silencing has offered a novel approach to the construction of new vaccines.Our study aimed to construct a recombinant influenza A ( H1 N1 ) virus with the PB1 gene that carries the target fragment of miRNA Let-7b. Methods After comparing the sequence of the A/Nanjing/108/2009 H1N1 viral fragments with that of Let-7b, we selected PB1 as the optimal gene sequence, inserted the Let-7b binding target gene into PB1, ligated the modified fragments with pDP 2000, and named the recombinant plasmids pDP-mu-PB1 and pDP-sclb-PB1, respectively.We co-transfected the MDCK and 293T cells with the recombinant and other seven plasmids and injected the supernatant into the allantoic cavity of the chickenembryo for virus propagation, followed by detection of the virus by hemagglutination ( HA) assay and measurement of the viral titer by TCID50 .We amplified the viral cRNA by RT-PCR and identified the viruses by agarose gel electrophoresis and nucleotide sequence analysis. Results PB1 was the optimal sequence ( 83 bp -107bp) for the attenuation of viruses.The HA-titers of miRT-H1N1 and scbl-H1N1 were 1∶32 and 1∶64, and their viral loads were 4.68 ×105 and 7.94 ×104 TCID50/mL, respectively.Nucleotide sequence analysis showed the expected fragment in the rescued virus. Conclusion A recombinant strain vaccine was successfully constructed, which has laid the foundation for fur-ther assessment of virulence.

ITS2 sequence was used as a barcode to identify Viticis Fructus and its adulterants. The study collected 46 samples include Viticis Fructus and its adulterants. The sequences were obtained by extracting DNA, amplification, sequencing bi-direstionlly, and then assembling with CodonCode Aligner. To identify Viticis Fructus and its adulterants, the study used the methods of computing the genetic distances by Kimura 2-Parameter (K2P) model and constructing the Neighbor-joining (NJ) phylogenetic trees using MEGA5.0. The results showed that the maximum inter-specific K2P distance of Viticis Fructus was less than the minimum inter-specific K2P distance of Viticis Fructus and its adulterants and the NJ tree indicated that the Viticis Fructus and its adulterants could be distinguished clearly. Therefore, using ITS2 barcode can distinguished Viticis Fructus and its adulterants accurately.

OBJECTIVE: To study the in vitro antibacterial activity of the crude extract of total flavonoids from Lonicera japonica against the main pathogenic bacteria.METHODS: With DM130 macroporous resin as stationary phase,the different components(component A contained more sugar,and content B less sugar;C,D and E components were eluted by 20%,40%,and 60% ethanol,respectively) were obtained by gradient elution of the aqueous extract from Lonicera japonica using different concentration of ethanol,then the MIC of different components on main pathogenic bacteria were detected.RESULTS:The antibacterial activity of component B against staphylococcus aureus was more ponent with its MIC at about 1 mg?mL-1.CONCLUSION:Component B has ponent antibacterial action on main pathogenic bacteria.

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