Objective To explore the impact of Th17 cells adoptive immunity on tumor growth of diffuse large B cell lymphoma (DLBCL)-bearing mice.Methods The CD4+ CD62L+ na(i)ve T cells purified by Mini MACS immunomagnetic beads were stimulated under the condition of TGF-β, IL-6, anti-IFN-γ, anti-IL-4, IL-23.IL-17 expression was detected by ELISA.SCID mice were inoculated with DLBCL cells line SUDHL-4 to estaldish the model of DLBCL-bearing mice, and then they received Th17 cells adoptive immunotherapy at the 0, 8 th or 18th day (3 groups in total, 5 mice pre-group).The tumor volume and the survival of tumor model were observed.Results Formation of tumor nodules in mice was observed at about the 8th day after the mice received the Th17 SUDHL-4 cells.The tumors volume of DLBCL-bearing mice that had received Th17 cells adoptive immunotherapy was obviously smaller than that of the mice had not (P ＜ 0.05).The survival time of the tumor-bearing mice that had received Th17 cells adoptive immunotherapy was also longer than that of the tumor-bearing mice had not (P ＜ 0.05).Conclusion Th17 cells adoptive immunotherapy displays an anti-tumor effect on DLBCL-bearing mice.

Objective To explore the effect of recombinant human granulocyte colony stimulating factor (rhG-CSF) mobilization on TH17/Treg cells and its impact on suppressor of cytokine signaling-3 (SOCS3) gene expression in CD4+ T cells in donors' peripheral blood.Method Sixteen donors were injected subcutaneously with rhG-CSF 5 μg/kg every day for 5 consecutive days for peripheral blood stem cells mobilization.At the first 0,3,5 day,the mononuclear cclls (MNCs) in peripheral blood or graft and serum specimens were taken.The CD4 + T cells in MNCs were sorted using immuno-magnetic beads.The ratio of TH 17 and Treg cells in MNCs,cytokines concentrations of IL-17A,IL-21,ID23 and TGFβ1 in serum,and SODC3 gene expression in CD4+ T cells were detected by using flow cytometry,ELISA,and reverse transcription real-time quantitative PCR (RT-qPCR),respectively.Results (1)The ratio of Th17 cells (CD3+ CD8 CD17+) and Treg cells (CD4+ CD25+ Foxp3+) in MNCs in peripheral blood and graft at the first 0,3 and 5 days after mobilization was (2.69 ± 0.81) ％,(0.91 ± 0.33) ％,(0.35 ± 0.12) ％,(0.21 ± 0.05) ％,and (0.56 ± 0.24) ％,(0.72 ± 0.22％),(1.59 ± 0.54) ％,(3.38 ± 0.52) ％,respectively,showing a significant declining and increasing trend respectively (P＜0.05); (2)The cytokine concentrations in serum at the first 0,3 and 5 days after mobilization were 7.33 ± 0.89,5.78 ± 1.03 and 3.32 ± 0.84 μg/L for IL-17A; 124.56 ± 15.18,117.12 ± 14.45 and 64.94 ± 11.25 μg/L for IL-21 ; 183.52 ± 59.35,280.49 ± 69.75 and 393.62 ± 57.25μg/L for TGF-β1 (P＜0.01) ; and 45.89 ± 6.95,46.25 ± 7.44 and 47.45 ± 10.75 μg/L for IL-23,respectively.The IL-17A and IL-21 concentrations showed significant declining trend,contrarily TGF-β1 with an increasing trend,while IL-23 concentration had no change.After rhG-CSF mobilization,the SOCS3 gene expression in CD4 + T cells of peripheral blood and graft at the first 0,3,5 days was gradually increased.Conclusion rhG-CSF suppresses Th17 cells and promotes regulatory T cells generation,meanwhile decreases IL-17A and IL-21 and elevates serum TGF-β1 concentrations,and contributes to CD4 + T cells differentiation to Tregs,probably by elevating SOSC3 gene expression in CD4+ T cells.

Objective To investigate the relationship between the helper T cell 17 (TH 17)/ Regulatory T cells (Treg cells) balance in peripheral blood with acute graft-versus-host reaction (aGVHD) after allogeneic hematopoietic stem cell transplantation (allo-HSCT),as well as the impact of anti-thymocyte immunoglobulin (ATG) on helper T cells in peripheral blood.Method Seventyeight hematologic patients underwent allo-HSCT,conditioning with or without ATG.Ten healthy volunteers severed as a control group.The helper T and regulatory T cells in peripheral blood were detected by flow cytometry.Enzyme-linked irnmunosorbent assay (ELISA) was used to detect serum concentrations of interleukin(IL)-17,IL-21,IL22,IL23,γ interferon (IFN-γ),and transforming growth factor β1 (TGF-β1).Result The percentage of Treg cells,TH17 cells and ratio of TH17/Treg cells in patients without aGVHD showed no significant difference from the healthy controls (P＞ 0.05).As compared with control group and non aGVHD group,the ratio of Treg cells was increased,the percentage of TH 17 cells,and TH 17/Treg cells were significantly increased in 1-2-degree aGVHD group (P＜0.01).With increased degree of aGVHD,the difference as above was more significant in 3-4-degree aGVHD recipients (P＜0.01).In aGVHD group,the IL-17,IL-23,IL-21 and IFN-γ concentrations were higher than the healthy group (P＜0.01) and non-aGVHD group (P＜0.05).Serum TGF-β1 level in aGVHD group was significantly decreased as compared with healthy group and non-GVHD group (P＜0.05),while IL-22 concentrations showed no statistically significant difference among three groups (P＞0.05).In anti-thymocyte immunoglobulin (ATG) pretreatment group,the absolute count of peripheral blood lymphocytes was less than in healthy control group (P＜0.01).In ATG group,the absolute counts of TH1 cells,TH17 cells,CD3+ CD4+ cells and non-TH1/17 cells were less than in non-ATG group (P =0.0000),while the absolute counts of lymphocytes,CD3+ CD4-cells,and TH 1/17 cells were less than in non-ATG group,but there was significant difference (P＞0.05).Conclusion The balance of TH 17/Treg cells and related cytokines were closely associated with aGVHD after allo-HSCT,and ATG influences the reconstruction of TH 17 and Th1 cells at early stage.

Objective To analyze setup errors and guide the calculation of margins from clinical target volume (CTV) and planning target volume (PTV) in esophageal cancer patients treated with tomothcrapy by the MVCT image-guided system.Methods Sixty-four esophageal canccr patients trcated with tomotherapy in our hospital in 2016 were randomly selected.MVCT images were acquired after patients' positioning and co-registered with KVCT images.The setup errors of x,y,and z translations and roll rotation were analyzed with the t-test or one-way ANOVA.Meanwhile,PTV margin was calculated based on the formula of M =2.5 Σ + 0.7δ Results According to the formula,the CTV-PTV margins in the x,y and z directions are slightly different between cancers located in the cervical,upper thoracic,middle thoracic,and lower thoracic segments.In patients with upper thoracic esophageal cancer,the average setnp error in the yaxis was lower when the head-neck-shoulder thermoplastic film fixation was used than when somatic thermoplastic film fixation (P=0.000);the setup errors of z-axis with somatic thermoplastic film fixation in the fifth and sixth weeks were slightly less than those in the first several weeks (P =0.036);the setup errors acquired by three image registration patterns were similar (x-axis P=0.868,y-axis P=0.491,z-axis P=0.169,roll P=0.985).Conclusions In the treatment of patients with esophageal cancer,the setup errors are large,but the MVCT in the TOMO HD system can greatly reduce the setup errors,ensuring the accuracy of each treatment.It is further recommended that in clinical practice,different CTV-PTV margins should be used for the treatments of esophageal cancers located in different segments.Patients with upper thoracic esophageal cancer are advised to use the head-neck-shoulder thermoplastic film fixation.

ObjectiveTo investigate the effect of artesunate on the expression of vascular endothlial growth factors(VEGF)and VEGFR in SHI-1 cell line.MethodsEnzyme-linked immunosorbent assay analysis was performed to detect the amount of VEGF in culture supernatants of SHI-1 cell in the condition of artesunate or not. The expression of VEGFR1 and VEGFR2 in SHI-1 cell in the condition of artesunate or not were detected by flow cytometry.ResultsWithout artesunate,the concentration of VEGF in the culture supernatant of SHI-1 cell were (980.3±2.2) pg/ml in 24 h and (982.4±2.3) pg/ml in 48 h. The expression of VEGFRI in SHI-1 cell were (6.40±3.11) ％ in 24 h and (6.45±2.85) ％ in 48 h. The expression of VEGFR2 in SHI-1 cell were (13.90±2.26) ％ in 24 h and (13.95±1.96) ％ in 48 h. With artesunate at 5, 10, 20 ng/ml, the concentration of VEGF in culture supematant of SHI-1 cell were (234.6±1.8)pg/ml, (114.9±1.6)pg/ml, (108.8±1.5) pg/ml in 24 h and (62.3±1.7) pg/ml, (60.9±1.6) pg/ml, (32.7±1.7) pg/ml in 48 h, respectively. The levels of VEGF in SHI-1 cells treated with artesunate at different concentrations decreased significantly (P ＜0.05).There was significant difference between 24 hours group and 48 hours group(P ＜0.05).The expression of VEGFR1 in SHI-1 cell were (4.30±2.21) ％, (4.20±1.37) ％, (3.90±1.86) ％ in 24 h and (3.80±2.87) ％, (3.60±1.73) ％, (3.00±1.82) ％ in 48 h, respectively. The expression of VEGFR1 in SHI-1 cell treated with artesunate at different concentrations were not significantly different (P ＞0.05). No significant difference between 24 hours group and 48 hours group was observed (P ＞0.05). VEGFR2 expression of SHI-1 cell were(4.40±1.15) ％, (3.10±0.68) ％, (1.10±0.72) ％ in 24 h and (3.00±1.68) ％, (2.20±0.93) ％, (0.60±0.92) ％ in 48 h, respectively. The results indicated that the expression of VEGFR2 in SHI-1 cells treated with artesunate at different concentrations reduced significantly (P ＜0.05),but there was no significant difference between 24 h group and 48 h group (P ＞0.05). ConclusionThe concentration of VEGF in SHI-1 cell was high, and artesunate can down-regulate the expression of VEGF and VEGFR2,but the effect of artesunate on the VEGFR1 was not significant.

Objective To evaluate the effectiveness and side effect of MT regimen (mitoxantrone plus teniposide) in inductive chemotherapy and explore the relationship between the effectiveness and karyotype. Methods 33 patients with acute monocytic leukemia were divided into two groups according to the treatment history or risk status according to cytogenetics MRC criteria. Group A (n=23) and B (n=10) were primary treatment and no remission following one course of DA (daunorubicin plus cytarabine) or HDA (Harringtonine,daunorubicin plus cytarabine) regimen,respectively. According to MRC criteria,group C (n=29) and D (n=4) were intermediate and adverse group. All the cases received two courses MT regimen chemotherapies to induce remission. The results and side effects were analysed. Results The complete remission rate and effective rate in group A and B were 83 % (19/23) and 60 % (6/10),91 % (21/23) and 70 % (7/10) respectively. The complete remission rate and effective rate in group C and D was 83 % (24/29) and 25 % (1/4),88 % (26/29) and 50 % (2/4) respectively. In complex cytogenetic group and 11q23 abnormal without complex cytogenetic group,CR rate was 0 (0/3) and 100 % (4/4). The time point,count of WBC nadir and the duration of WBC were less than 1×109/L is (7±3) day after chemotherapy,(0.4±0.2)×l09/L,(8±5) day. Chemotherapy related mortality was 0. Conclusion MT regimen was highly effective and safe in inducing remission in acute monocytic leukemia,including the cases which achieved no remission following one course of DA or HDA regimen. The effectiveness of MT regimen relates to the cytogenetics. MT regimen may be highly effective in cases with 11q23 abnormal and poor effective in cases with complex cytogenetic.

Objective To understand the changes of Th17 cells and related cytokines in diffuse large B-cell lymphoma (DLBCL) patients treated with rituximab and its significances.Methods Patients were assigned to 4 groups,there were 20 cases in the control group,31 cases in the initial treatment group,31 cases in CHOP group and 25 cases in RCHOP group.The percentage of Th17 cells in the peripheral blood of each group was tested by flow cytometry,the related cytokines IL-17,IL-21,IL-23,TGF-β in the peripheral blood were measured by enzyme-linked immunosorbent assay.Results The percentage of Th17 cells and the levels of IL-17,IL-21,IL-23 in the initial treatment group [(0.67±0.21) ％,(5.929±1.342) pg/ml,(130.632±17.945)pg/ml,(51.681±9.808) pg/ml] and the CHOP-CR group [(1.07±0.37) ％,(6.526±0.538) pg/ml,(132.119±7.700)pg/ml,(50.245±7.668) pg/ml] were both significantly lower than those in the control group[(2.53±0.63) ％,(8.435±2.031) pg/ml,(149.265±12.316) pg/ml,(55.303±7.778) pg/ml] (P ＜ 0.05).The level of TGF-β in the initial treatment group [(370.615±98.444) pg/ml] was significantly higher than that in the control group [(311.895±73.365) pg/ml] (P ＜ 0.05).The percentage of Th17 cells and the levels of IL-17,IL-21,IL-23 in the RCHOP-CR group [(2.38±0.59) ％,(7.724±0.780) pg/ml,(148.412±7.355) pg/ml,(55.668±7.532) pg/ml] were significantly higher than those in the initial treatment group [(0.67±0.21) ％,(5.929±1.342) pg/ml,(130.632±17.945) pg/ml,(51.681±9.808) pg/ml] and the CHOP-CR group [(1.07±0.37) ％,(6.526±0.538) pg/ml,(132.119±7.700) pg/ml,(50.245±7.668) pg/ml] (P ＜ 0.05).The level of TGF-β in the RCHOP-CR group[(283.904±59.223) pg/ml] was significantly lower than that in the CHOP-CR group [(341.481±95.597) pg/ml] (P ＜ 0.05).Conclusion Th17 cells might be negatively correlated with the DLBCL development,the reduced IL-23 and elevated TGF-β might suppress the differentiation of Th17 cells.Rituximab could elevate the percentage of Th17 cells in DLBCL patients,and it is related with the effect of chemotherapy.

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BACKGROUND: Vascular injuries of the extremitiesy are frequently concomitant with vascular defects and are usually repaired by autologous vessel grafting. However, the source of autologous vessels is limited and the preparation of them is traumatic.OBJECTIVE: To retrospectively evaluate the extremity-preserving efficacy of vascular prosthesis grafting in repairing major vascular injuries of the extremities in 29 cases.DESIGN: A retrospective study.SETTING: Department of Orthopedics and Traumatic Surgery, Center for Battle Wound and Trauma of Chinese PLA, Research Institute of Surgery,Daping Hospital, Third Military Medical University of Chinese PLA.PARTICIPANTS: We selected 29 patients with major vascular injuries of the extremities repaired by vascular prosthesis grafting who received the treatment at the Department of Orthopedics and Traumatic Surgery of the Center for Battle Wound and Trauma of the Chinese PLA of the Research Institute of Surgery of, Daping Hospital of the Third Military Medical University of Chinese PLA between January 1989 and December 2000. There were 23 males and 6 females. Injury sites: 9 cases were at subclavian artery, 6 at axillary artery, 2 at brachial artery, 10 at femoral artery, 1 at femoral vein and 1 at popliteal artery. 11 of these 29 cases (37.9%) were complicated with shock, 8 with fractures and dislocations, 5 with peripheral nerve injuries and 3 with infections.METHODS: The vascular prosthesis was anastomosed end to end with the trimmed culprit vessels. As for the 3 cases with infective arterial injuries,the vessels were placed away from the infected region and were bridged together in a non-inf1ammatory region and then muscles or musculo-cutaneous flaps were used to cover the infected regions.The functions of the extremities of the patients were evaluated according to MAS at week 2 and 1 year after the operation.MAIN OUTCOME MEASURES: The vascular patency rate and the extremity-preserving outcomes..RESULTS: All 29 patients entered the stage of result analysis. Extremitypreserving efficacy: All the patients who underwent vascular prosthesis grafting had their extremities preserved. Only 1 patient had the late sequelae of ulcers in the foot sole due to extended extremity ischemia and nerve damage. The rate of 2-week patency rate of the grafted vessels was 100% and the 1-year rate of patency was 96.5%, as revealed by Doppler blood stream scanning. The excellent and good rate of the function of extremity was 89%.CONCLUSION: Vascular prosthesis grafting is the one of the methods for repairing the major vascular injuries so as to preserve the extremities and their functions.

Objective To explore the impact of rituximab on Th17 cells and related cytokines in patients with diffuse large B-cell lymphoma (DLBCL) in vitro and its significance.Methods 20 cases of DLBCL untreated patients and 20 healthy subjects were enrolled in the name of DLBCL group and health control group, respectively.4 peripheral blood samples were collected from every case to separate peripheral blood mononuclear cells (PBMCs), which were assigned to 4 subgroups according to different culture conditions: blank subgroup(subgroup A), rituximab subgroup (subgroup B), rituximab and serum subgroup (subgroup C) and polarization subgroup (subgroup D) (added IL-6 and TGF-β).After cultured in vitro, the percentage of Th17 cells in each subgroup was tested by flow cytometry, and the cytokine IL-17 in the abovementioned culture fluid was measured by enzyme-linked immunosorbent assay (ELISA).Results In health control group, the percentage of Th17 cells and the level of IL-17 in subgroup D [(17.12 ± 4.90) ％ and (45.735±10.012) pg/ml] were significantly higher than those in subgroup A, B, C (P ＜ 0.05), and there was no difference in each other subgroup A, B, C (P ＞ 0.05).The percentage of Th17 cells and the level of IL-17 in the DLBCL subgroup A were significantly lower than those in health control subgroup A [(0.69±0.24) ％ and (6.012±1.312) pg/ml vs (2.43±0.61) ％ and (8.217±1.681) pg/ml (P ＜ 0.05)].In DLBCL group, after cultured with rituximab, the percentages of Th17 cells in subgroup B, C, D were (2.34±0.48) ％, (2.31±0.53) ％ and (16.92±4.81) ％, and the levels of IL-17 were (7.944±1.538) pg/ml, (7.957±1.533) pg/ml and (44.417±9.881) pg/ml, respectively, which were all significantly higher than those in subgroup A.Besides, the percentage of Th17 cells and the level of IL-17 in DLBCL subgroup D were significantly higher than those in subgroup B, C (P ＜ 0.05), while there was no difference between subgroup B and subgroup C.Conclusion Experiments in vitro confirmed that the percentage of Th17 cells in PBMCs of DLBCL patients was lower than that in healthy persons, and rituximab could elevate the percentage of Th17 cells in PBMCs of DLBCL patients.