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OBJECTIVE To study the vasorelaxant effects and mechanism of polyphenol compound 2，4′，5′-trihydroxy-5，2′- dibromo-diphenyl-methanone（LM49）on isolated aortic rings of rats. METHODS Thoracic aortic vascular rings of rats were collected. Using the diastolic rate as index ， the effects of different concentrations of LM 49 on endothelium-intact and endothelium-denuded aortic rings pre-contracted by norepinephrine （NE，1×10－6 mol/L）or KCl （60 mmol/L）were investigated. After pre-culturing vascular rings by nitric oxide synthase inhibitor L-nitro-arginine methyl ester （L-NAME，0.1 mmol/L） and cyclooxygenase inhibitor indomethacin （1×10－5 mol/L），as well as pre-culturing vascular rings by 4 potassium channel blockers [BaCl 2（0.1 mmol/L），tetraethylammonium（TEA，5 mmol/L），4-aminopyridine（4-AP，0.1 mmol/L）and glibenclamide （1×10－5 mol/L）]，the vasorelaxant effect of different concentrations of LM 49 on the vascular rings were investigated by using the same method. With the percentage of vasoconstriction as the index ，using KCl （60 mmol/L），NE（1×10－6 mol/L），calcium channel blocker verapamil （1×10－6 mol/L）and sarcoplasmic Δ 基金项目 重大新药创制国家科技重大专项 （No.2018ZX097- reticulum Ca 2 +-adenosine triphosphate （ATP） enzyme pump inhibitor thacarotene （TG，1×10－6 mol/L）to induce the release of calcium in vascular rings in the absence of calcium. CaCl was added cumulatively ，and the effect of LM 49 on the cxyw06，vasoconstriction caused by calcium influx induced by CaCl 2 was investigated. RESULTS 3×10－6，5×10－6，1×10－5 mol/L LM49 had a significant relaxation effect on NE and KCl precontracted vascular rings （P＜0.01）； whether the endothelium was removed or not had no significant effect on the vasodilation of LM 49（P＞0.05）. After L-NAME ，indomethacin， TEA and 4-AP was pre-incubated ，different concentrations of LM 49 had no significant effects on aortic rings precontracted by NE （P＞0.05）. Glibenclamide and BaCl 2 could inhibit the vasorelaxant effects of LM 49 on aortic rings precontracted by NE （P＜0.01）. In the absence of calcium ，LM49 could inhibit the contraction caused by calcium influx induced by accumulated CaCl 2 after pre-incubation with KCl ，NE，verapamil and TG （P＜0.01）. CONCLUSIONS LM49 evokes significant relaxation of isolated aortic vascular rings without endothelium dependence ；the mechanism of which is inducing ATP-sensitive potassium channel ， inward rectifier potassium channel open and restraining extracellular Ca 2 + influx via voltage-gated calcium channel ， receptor-operated calcium channel and store-operated calcium channel.
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Objective:To evaluate the effect of down-regulating lncRNA LINC00263 targeting miR-4458 on the proliferation, migration, invasion and radiosensitivity of breast cancer SK-BR-3 cells.Methods:The expression differences of LINC00263 in breast cancer tissues, adjacent tissues, normal breast epithelial cells and breast cancer cells were determined by qRT-PCR. Transfection of LINC00263 shRNA in breast cancer SK-BR-3 cells down-regulated the expression of LINC00263, and the cloning experiment was used to detect the radiosensitivity. Breast cancer SK-BR-3 cells were treated with 6 Gy irradiation. CCK-8 assay was employed to detect cell proliferation. Flow cytometry was adopted to detect cell apoptosis. Transwell chamber test was performed to detect cell migration and invasion. Western blot was used to detect the expression levels of C-Caspase-3 and C-Caspase-9, MMP-2 and MMP-9 proteins. Bioinformatics software predicted that LINC00263 and miR-4458 had complementary binding sites, and the luciferase reporter system was utilized determine the targeting relationship between LINC00263 and miR-4458. LINC00263 shRNA and miR-4458 inhibitor were co-transfected into breast cancer SK-BR-3 cells, and 6 Gy irradiation was given to detect the changes in cell proliferation, apoptosis, invasion and migration.Results:The expression level of LINC00263 in breast cancer tissues was higher than that in adjacent tissues. The expression level of LINC00263 in breast cancer cells was higher compared with that in normal breast epithelial cells. The radiosensitivity of breast cancer SK-BR-3 cells was increased after transfection of LINC00263 shRNA. Transfection of LINC00263 shRNA and radiation exerted a synergistic effect, jointly inhibited breast cancer cell proliferation, migration and invasion, promoted cell apoptosis, up-regulated the expression levels of C-Caspase-3 and C-Caspase-9 proteins in cells, and down-regulated those of MMP-2 and MMP-9 proteins. Down-regulation of LINC00263 targetedly up-regulated miR-4458 expression. miR-4458 inhibitor reversed the inhibitory effect of LINC00263 shRNA combined with radiation on the proliferation, migration, invasion and apoptosis promotion of breast cancer SK-BR-3 cells.Conclusion:Down-regulating lncRNA LINC00263 targeting miR-4458 inhibits the proliferation, migration and invasion of breast cancer SK-BR-3 cells, and improves cell radiosensitivity.
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Irreversible electroporation (IRE) is an emerging tissue ablation technique. Compared with thermal ablation technique such as radiofrequency, IRE can achieve focal ablation in a shorter time without heat sink effect while sparing the tissue scaffold. IRE has been demonstrated to be a feasible therapeutic modality for the liver, pancreatic, and prostatic cancer. In recent years, several studies regarding of catheter-directed IRE for digestive tract, bronchus, urinary tract, and myocardium have been performed, which preliminarily demonstrated the safety and efficacy of IRE for tissue ablation under endoscopic or interventional technique. This study summarized the research progress of catheter-directed IRE for tissue ablation. The critical technique and future direction of catheter-based IRE are prosp.
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Catheter Ablation , Catheters , Electroporation , Endoscopy , HumansABSTRACT
Early diagnosis of pancreatic cancer remains difficult, and it progresses rapidly. Only a small part of patients are eligible for initial resection at the time of diagnosis, most patients need to benefit from palliative treatment. As a kind of palliative treatment, physical therapy aims to control the growth of primary tumors, relieve patients′ symptoms and improve their quality of life. In recent years, with the development of basic science, the physical treatment of tumors has also been continuously innovated. It has become an important part of comprehensive treatment measures for pancreatic cancer, and is of great significance for improving the prognosis of pancreatic cancer patients. This article summarizes the physical therapy methods related to the treatment of pancreatic cancer, including radiofrequency ablation and radiotherapy, etc.
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Objective To study the significance of Th17 cells in patients with myelodysplastic syndrome (MDS) and iron overload.Methods A total of 77 patients with MDS admitted to Guangzhou First People's Hospital were enrolled from January 2017 to December 2018,who were divided into iron overload group (37 cases) with serum ferritin (SF) over 1000 μg/L and non-ferrous overload group(40 cases).CD4+T cells in peripheral blood (PB) and bone marrow (BM) were sorted by flow cytometry.The ratio of Th17 cells and cells with abnormal karyotype were compared.IL-17 and IL-6 protein and RNA expression were detected by ELISA and quantitative real-time PCR(qRT-PCR).Results The proportions of Th17 cells in PB and BM in iron overload group were significantly higher than those in non-iron overload group [(41.06± 0.96)％ vs.(26.80± 1.21)％;(47.39± 1.60)％ vs.(34.29± 1.03)％;P＜0.01].The Th17 positive cells with abnormal karyotype in iron overload group were more than those in non-iron overload group[(4.96±0.53)％ vs.(3.67±0.12)％ in PB;(10.06±1.67)％ vs.(4.36±0.43)％ in BM;P＜0.01].Similarly,the protein levels as well as mRNA expression of IL-6 and IL-17 in patients with iron overload were significantly higher than those in non-iron overload group (P＜0.01 both in PB and BM).Conclusions As hematopoietic regulators secreted by Th17 cells,the expression of IL-6 and IL-17 in MDS patients with iron overload are elevated.This may predict the influence of these factors to the differentiation of Th 17 cells.
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Objective To investigate the effects RORrt (RORrt),IRF8 (IRF8) and STAT3 (STAT3) in peripheral blood CD4+T cells on the cell proliferation and differentiation in elderly patients with iron-overload myelodysplastic syndrome(MDS).Methods A prospective case-control study was conducted.Twenty-two elderly hospitalized patients(12 males and 10 females)aged 60-78 years with iron-overload MDS from Jan.2017 to Dec.2018 were enrolled and considered as the observation group.Twenty MDS patients without iron overload hospitalized in the same period were selected as the non-iron overload group,and 26 healthy elderly people were considered as the healthy control group.Peripheral blood monocytes(PBM)were prepared and resident CD4+T cells were sorted by flow cytometry.The mRNA and protein expression levels of transcription factors of RORrt,p-STAT3 and IRF8 were detected by quantitative real time polymerase chain reaction(qRT-PCR)and Western blotting.Results In peripheral blood CD4+T cells,the mRNA expression level of RORrt and p-STAT3 were higher and that of IRF8 was lower in the iron-overload group than in the non-iron overload group and the healthy control group(42.634± 18.613 vs.21.289 ± 15.158 and 22.520 ± 9.896;29.710±9.689 vs.12.355±4.681 and 9.818±3.845;19.293±8.258 vs.23.785±12.498 and 69.619±23.768,P＜0.01).In peripheral blood CD4+T cells,the protein expression level of RORrt and p-STAT3 was higher,and that of IRF8 was lower in the iron overload group than in the non-iron overload group and healthy control group(P＜0.01).Conclusions The abnormalities of the mRNA and protein expression levels of transcription factors of RORrt,IRF8 and p-STAT3 in CD4+ T cells play a fundamental role in the pathogenesis of iron overload MDS in elderly patients.
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Objective To investigate the effects of high intensity electric field on cell growth,apoptosis and microstructure of human pancreatic cancer cell line PANC-1.Methods The PANC-1 cells in the logarithmic growth period were selected,and cells in the high voltage electrical treatment group were treated with high voltage electric field 250,500,750,1000 V/cm,respectively.The effects of different high voltage electric fields on cell growth and microstructure of PANC-1 cells were determined by cell viability,cell death staining,apoptosis detection,transmission electron microscopy and scanning electron microscopy.Results Compared with control group,the high voltage electric pulse significantly inhibited the growth of PANC-1 cells in the field dependent manner.Moreover,when the field was more than 500 V/cm,the cell viability was significantly decreased (P＜0.05).High voltage electric pulse could induce cell apoptosis.When the field was higher than 750 V/cm,serious necrosis was noticed.In the 1000 V/cm group,the integrity of cell membrane and the structure of organelles was seriously damaged.Conclusion High voltage electric pulse could significantly inhibit the growth of PANC-1 cells and would be a promising method in cancer treatment.
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Objective To analyze the risk factors of early acute kidney injury (AKI) after liver transplantation from donation after cardiac death(DCD) donor liver. Methods Clinical data of 184 donors and recipients undergoing liver transplantation from DCD donor liver were retrospectively analyzed. According to the incidence of early AKI, all participants were divided into the AKI and non-AKI groups. The patients in the AKI group were subject to AKI staging. Baseline data, preoperative, intraoperative and postoperative related parameters were statistically compared between two groups. The cumulative survival rate and clinical prognosis of patients in non-AKI group and AKI group with different staging were statistically analyzed by Kaplan-Meier curve analysis. Results Among 184 patients, 68 cases (37.0%) presented with early AKI after liver transplantation including 31 stage 1 AKI, 26 stage 2 AKI and 11 stage 3 AKI, mainly occurring within postoperative 3 d. Univariate analysis revealed that preoperative levels of albumin <35 g/L, preoperative levels of serum sodium ≤137 mmol/L, operation time>7.5 h, intraoperative hemorrhage volume>3 000 mL, intraoperative red cell infusion volume>15 U and intraoperative urine amount ≤100 mL/h were the risk factors of early AKI after liver transplantation (all P<0.05). Multi-variate Logistic regression analysis demonstrated that intraoperative red cell infusion >15 U was an independent risk factor of early AKI after liver transplantation [odds ratio(OR) 1.061, 95% confidence interval(CI)1.008-1.118,P=0.024].Result of Kaplan-Meier survival curve suggested that the cumulative survival rate was gradually reduced along with the aggravation of AKI with statistical significance (all P<0.05). Conclusions The incidence of early AKI following liver transplantation is relatively high. The severity of early AKI is intimately correlated with the short- and long-term prognosis of the recipients. A large quantity of intraoperative red blood cell infusion is an independent risk factor of AKI.
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Objective To investigate the correlation between histone deacetylase 1(HDAC1) and vascular endothelial growth factor(VEGF) in the patients with lung adenocarcinoma.Methods Eighty cases of lung adenocarcinoma in our hospital from August 2014 to April 2016 served as the research subjects,and contemporaneous 80 individuals undergoing healthy physical examination were taken as the control group.The fasting venous blood sample was collected in all subjects.Then serum HDAC1 and VEGF levels were detected by ELISA.The differences of serum HDAC1 and VEGF expression levels were compared between the two groups.The HDAC1 and VEGF expression levels in the patients with different characteristics of lung adenocarcinoma and the relation between serum HDAC1 and VEGF concentrations were analyzed.Furthermore the possible influence factors of HDAC1 protein expression level in the patients with lung adenocarcinoma were analyzed.Results The HDAC1 levels in the control group and observation group were(329.56 ± 23.83) ng/L and(568.20 ± 35.40) ng/L,the difference was statistically significant(t=23.576,P=0.000).The VEGF levels in the control group and observation group were(40.26±9.82)ng/L and(296.56±19.80)ng/L respec tively,the difference was statistically significant(t=31.154,P=0.000).The HDAC1 protein level had statistical difference among different genders,ages,clinical stages and smoking history,the HDAC1 protein level in male,age ＞60 years old,clinical stage Ⅲ,V and patients with smoking history were higher(P＜0.05).The Pearson correlation analysis results showed that serum HDAC1 in the patients with lung adenocarcinoma was positively correlated with VEGF protein concentration(r=0.526,P =0.000).The Logistic regression analysis showed that influence factor of HDAC1 protein expression level in the patients with lung adenocarcinoma was clinical stage.Conclusion The high expression of HDAC1 protein in lung adenocarcinoma patients may also simultaneously regulate the VEGF expression,thus promotes the development of lung adenocarcinoma.
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Objective: To investigate the efficacy and safety of using pegylated recombinant human granulocyte-colonystimulating factor (PEG-rhG-CSF) in preventing neutropenia in multiple chemotherapy cycles. Methods: A multicenter, prospective, open-label, singlearmstudy was designed. Patients with malignant tumors, such as lung, ovarian, and colorectal cancers, who received multiple cycles of chemotherapy with the prophylactic use of PEG-rhG-CSF for 2-4 consecutive cycles participated in the study. Results: After the prophylactic use of PEG-rhG-CSF, the incidence of grade IV neutropenia decreased from 4.76% (13/273) in the first cycle to 1.83% (5/273), 1.15% (2/174), and 2.08% (2/96) in subsequent cycles. Meanwhile, the incidence of grade III neutropenia decreased from 11.36% (31/ 273) in the first cycle to 6.23% (17/273), 2.87% (5/174), and 3.13% (3/96) in subsequent cycles. The incidence of febrile neutropenia (FN) during the first cycle was 0.73% (2/273). The duration of FN was 2 days in one case and 5 days in another case. FN was not observed during the second, third, or fourth cycle. After the secondary prophylactic use of PEG-rhG-CSF, the incidence of grade IV neutropenia decreased from 25% (7/28) to 3.57% (1/28), 0% (0/28), and 6.67% (1/15) in subsequent cycles. Meanwhile, the incidence of grade III neutropenia decreased from 71.43% (20/28) to 10.71% (3/28), 14.29% (4/28), and 0% (0/15) in subsequent cycles. The proportion of patients who received antibiotic therapy during the entire chemotherapy period was 10.48% (44/420). Conclusion: The application of PEG-rhG-CSF once per chemotherapy cycle can effectively reduce the occurrence of neutropenia in patients under multiple cycles of chemotherapy treatment with good safety.
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Objective To establish a mice model of diffuse large B-cell lymphoma (DLBCL) that was treated with adoptive immunity of Th17 cells cultured in vitro,and to analyze the relationship between IL-17 and MHC Ⅱ expression and their relation with tumor growth.Methods The CD4+CD62L+ T cells purified by MACS were stimulated under cytokine conditions including anti-CD3,anti-CD28,TGF-β and IL-6 in vitro,and SUDHL-4 cells were cultured and inoculated the SCID mice to establish DLBCL mice models.The mice were divided into Th17 cells immunity group (30 mice) and control group (20 mice).Th17 cells were injected to mice to get the adoptive immunity in immunity group,and 0.9 ％ NaCl in control group.The half mice were terminated at median disease onset time and median survival time,respectively.ELISA was used to detect IL-17 expression,and immunohistochemistry was applied to detect MHC Ⅱ expression in the tumor tissues.Results The median disease onset time of DLBCL mice model was 8 d,and median survival time was 28 d.The IL-17 and MHC Ⅱ expression levels in Th17 cells immunity group [(11.93±0.56) pg/ml,(69.13t0.36) ％] were higher than those in control group [(9.82±0.26) pg/ml,(42.59±0.12) ％] (both P＜ 0.000 1).Along with the progress of DLBCL,IL-17 and MHC Ⅱ expression levels were decreased [(9.53±0.18) pg/ml,(54.63±0.45) ％,both P ＜ 0.000 1].There was a significantly positive correlation between IL-17 and MHC Ⅱ (r=0.89,P=0.000).Conclusions The expression level of MHC Ⅱ can be used as a factor to judge the disease situation of DLBCL,and combination detection of the expression of both IL-17 and MHC Ⅱ will provide more reference values for judgment of the disease situation and the progress of DLBCL.
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<p><b>OBJECTIVE</b>To explore the effect of false positive signals during detection of BCR/ABL fusion gene by fluorescence in situ hybridization (FISH), and develop a method for calibration.</p><p><b>METHODS</b>Normal specimens were mixed with BCR/ABL positive specimens in which presented signal pattern of 1-red-2-green-1-fusion (1R2G1F) using dual color dual fusion (DCDF) probes and 1-red-1-green-1-fusion (1R1G1F) using extra signal (ES) probes in different proportions. Mixed samples were detected using DCDF and ES probes. Results of DCDF probes, ES probe before calibration, ES probes after calibration and theoretical results were compared by binomial distribution in different proportions.</p><p><b>RESULTS</b>The rate of false positive signals has risen with increase of negative rate. A significant difference was found between theoretical proportion and results without calibration in negative level, 5%, 10% and 25% positive level (P<0.05). There was no significant difference between theoretical proportion and results without calibration in 50% and 90% positive level (P>0.05). Also there was no significant difference between theoretical proportion and calibrated results (P>0.05).</p><p><b>CONCLUSION</b>Calibration of FISH result can delimitate the effect of false positives, and can provide more reliable results in cases with low level positive rates.</p>
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Adult , Calibration , False Positive Reactions , Female , Fusion Proteins, bcr-abl , Genetics , Humans , In Situ Hybridization, Fluorescence , Methods , Reference Standards , Leukemia, Myelogenous, Chronic, BCR-ABL Positive , Diagnosis , Genetics , Young AdultABSTRACT
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.
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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.
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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.
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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.
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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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Objective To investigate the pharmacokinetic difference between the combination of cyclophosphamide and cisplatin and single medication.Methods High performance liquid chromatography (HPLC) was used to detect the pharmacokinetic parameters and distribution parameters in tissue of cisplatin and cyclophosphamide.Results The pharmacokinetic curve of the CP regimen showed two-compartment model,and there was no significant difference between the absorption half life of the combination of the two drugs (0.56±0.02,0.72±0.02) and that of single medication (0.92±0.02,1.16±0.12) (t =2.091,t =2.379,both P ＞ 0.05).While the elimination half life of the combination (0.56±0.02,0.72±0.02) was significantly longer compared with that of single medication (0.92±0.02,1.16±0.12) and there were significant differences (t =5.796,t =6.213,both P ＜ 0.05).There was no significant difference of MRT in heart between the disturbution of the combination of the two drugs and that of single medication (t =2.231,t =2.096,both P ＞ 0.05).While there was significantly longer compared with that of single medication and there were significant differences in lung and liver (t =5.976,t =6.270,both P ＜ 0.05).Conclusion When used in combination,the elimination of the drugs is slower,which suggests that the interval and dosage of the drugs should be adjusted to their pharmacokinetic parameters in clinical use to improve effects and lower side effects.
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Objective To explore clinical features of chemotherapy-induced graft-versus-host disease and to observe the dynamical changes of Th17/Treg ratio and related cytokines in one relapsed acute T-lymphoblastic leukemia after allo HSCT.Methods Twenty health volunteers were healthy controls.One patient achieved complete haematologic remission after two courses of chemotherapy and underwent matched HLA identical sibling allogenic peripheral blood stem cell transplantation,donor cell implanted state,disease recurrence and graft-versus-host disease were observed and Th17/Treg cells and its related factors in the peripheral blood were detected in different periods dynamically changes by methods of flow cytometry and ELISA.Results The patient achieved complete donor chimerism (FDC) at day +27 after transplantation.Hematopoietic function was fully recovered at day +34.Chronic GVHD (cGVHD) occurred at day +110.Thereafter,extramedullary relapse occurred in cGVHD state at day +264.After one course of Hyper-CVAD chemotherapy,patient complicated overlap syndrome with cGVHD of oral cavity ulcer and degree Ⅳ intestinal aGVHD at day +294 and day +347,respectively.Th17 cell ratio in CD+4 T cells (1.7 ％) in the overlap syndrome slightly increased,compared with control group [(0.56±0.35) ％].The ratio of Treg cells in CD+4 T cells (4.66 ％) with cGVHD increased compared with normal control group [(0.59±0.33) ％],recurrence (0.39 ％),and hematopoietic stem cell implantation (1.15 ％),but the ratio of Treg cells increased significantly when patient complicated overlap syndrome (7.83 ％).The ratios of Th17/Treg were more than 1 at relapse stage and less than 1 at GVHD stage.The IL-17A level in serum significantly increased in cGVHD (6.114 pg/ml)and overlap syndrome (6.805 pg/ml) stage compared with normal control group [(5.19±0.77) pg/ml].TGF-β1 levels were significantly higher at different periods after transplantation compared with control group [(48.81±4.9) ng/ml].Conclusion Chemotherapy can induce GVHD in relapsed acute leukemia patients after hematopoietic stem cell transplantation,and the imbalance of the Th17/Treg cells and its cytokines maybe related with disease relapse and GVHD.
ABSTRACT
ObjectiveTo explore the relationship between the international prognosis indexes(IPT) and the cell-mediated immunity condition in DLBCL patients. Methods52 DLBCL patients were divided into 4 groups and the FCM was used to examine the T lymphocyte subsets,including CD3+、CD4+、CD8+、NK cells.T lymphocyte subsets absolute value and CD4+/CD8+ ratio were examined. Correlation with IPI were compared among groups.ResultsCD3+ cells in high risk group [(1570.9±370.5)/μl]were higher than other IPI groups;CD4+ cells in DLBCL groups were all lower than normal group(751.3±367.4)/μl]; CD8+ cells in high risk group [(1055.9±523.8)/μl] were higher than other groups; CD4+/CD8+ ratio in middle-high risk group and high risk group (1.0±0.2、0.7±1.0)were lower than other IPI groups and normal group;NK cells in middle-high risk group and high risk group were lower than the normal group[(199.5±68.4)/μl、(171.9±126.9)/μl];Age,clinical stage,body state had correlated with the CD3+ 、CD8+ cells and CD4+/CD8+ ratio of the DLBCL patients' peripheralblood T lymphosyte subsets. ConclusionsThe immunity condition in DLBCL patients has correlated with IPI; With increasing IPI value,the immunity depression and disorder become more serious,NK cells function become worse,and the prognosis is bad too.