Effects of early application of thymosin peptide alpha 1 on T lymphocyte subsets after hepatectomy in patients with hepatocellular carcinoma

AIM: To investigate the effects of early application of thymosin peptide alpha 1 on lymphocyte subsets after operation in patients with hepatocellular carcinoma. METHODS: Forty-six patients with hepatocellular carcinoma were randomly divided into control and treatment groups for this study. Thymosin α1 at dose of 1.6 mg was injected subcutaneously on day 1, 3, and 5 after operation in treatment group. The percentages of CD3+, CD4+ and CD8+ cells, and CD4+/CD8+ ratio in both groups were counted before operation and on day 1, 4, and 7 after hepatectomy. RESULTS: CD4+ cell population and CD4+/CD8+ ratio decreased, but CD8+ increased after operation in control group (P<0.05). In thymosin peptide alpha 1 treatment group, there was no statistical difference in the percentages of CD3+, CD4+, CD8+, and CD4+/CD8+ before and after operation. In addition, thymosin α1 significantly increased CD4+ cell population and CD4+/CD8+ ratio (P<0.05). CONCLUSION: Operation suppresses the immune function in patients with hepatocellular carcinoma. Thymosin α1 increases CD4+ T lymphocyte subsets in patients after operation.     

黄伞多糖对竞技运动员抵抗能力的作用机理

为了研究黄伞多糖对竞技运动员抵抗能力的作用机理,选择专业男子篮球运动员20人,任意划分成实验组与对照组。将子实体粉作为基础材料,进行粗多糖提取和提纯处理,制成黄伞多糖胶囊。令实验组运动员在实验期间每天早晚服用黄伞多糖胶囊,每次2粒,共服用35 d。对照组运动员除不服用黄伞多糖胶囊外,其余与实验组运动员相同。研究黄伞多糖对免疫球蛋白、T淋巴细胞亚群和NK细胞活性、IL-2水平的作用机理。结果表明,竞技运动员训练后和训练前相比,实验组IgA、IgG以及IgM明显增加,对照组IgA、IgG以及IgM明显降低,实验组IgA、IgG以及IgM非常显著性高于对照组;实验组运动员训练后CD4+百分和CD4+/CD8+值相比显著升高,CD8+百分比在一定程度上有所降低,训练后实验组运动员CD4+百分和CD4+/CD8+值显著高于对照组,CD8+百分比显著低于对照组;实验组与对照组NK水平训练后较训练前均有显著变化,训练后实验组较实验前有显著变化, IL-2水平在实验过程中一直无显著变化。得出以下结论:长期高强度训练能令竞技运动员抵抗能力降低,黄伞多糖可有效提高竞技运动员抵抗能力,其作用机理为提高CD4+百分和CD4+/CD8+值、T淋巴细胞亚群水平以及NK细胞活性。     

Elevation of blood Th17 cells and CD3+CD8-IL-21+ T cells in patients with cervical cancer

AIM: To investigate the role of Th17 cells in the patients with cervical cancer.METHODS: We measured the peripheral levels of Th17 cells and CD3⁺CD8^-IL-21⁺ T cells in 37 cervical cancer (CC) patients, 25 cervical intraepithelial neoplasia (CIN) patients and 18 healthy controls by flow cytometry. The percentages of Th17 cells and CD3⁺CD8^-IL-21⁺ T cells in total CD4⁺ cells were calculated.RESULTS: Compared with controls, the patients with CC or CIN had higher proportions of Th17 cells (all P<0.01) and CD3⁺CD8^-IL-21⁺ T cells (all P<0.05). Notably, in CC patients, the increased percentages of Th17 cells and CD3⁺CD8^-IL-21⁺ T cells were independently associated with the clinical stage(all P<0.05), lymph node metastasis (P<0.01,P<0.05) and vasoinvasion (all P<0.01), while the elevated percentage of CD3⁺CD8^-IL-21⁺ T cells was also associated with the tumor size(P<0.01). Furthermore, the percentage of Th17 cells was positively correlated with that of CD3⁺CD8^-IL-21⁺ T cells in healthy controls and CC patients, but not in CIN patients.CONCLUSION: Our results indicates a possible role of Th17 cells in CC patients correlated with CD3⁺CD8^-IL-21⁺T cells, and the elevated percentage of circulating Th17 cells may be involved in the development and progression of cervical cancer.     

Effect of lupus recipe on immune system and lymphocyte subsets proliferation in splenic cells of lupus-prone BXSB mice

AIM: To investigate the effects of lupus recipe on immune system and lymphocyte subsets proliferation in splenic cells in BXSB mice. METHODS: Eighteen male BXSB mice model was used in the experiment. The model mice were divided into three groups: un-treated model group, lupus recipe (LR) treated group, and prednisone treated group. All model mice were killed in 10 weeks. The control group consisted of 6 syngeneic normal C57BL/6 male mice. The levels of total IgG and anti-dsDNA antibody in serum were detected by ELISA. The percentages of lymphocyte subsets (CD3+, CD4+, CD8+ T lymphocytes and CD19+, CD23+ B lymphocytes) were detected by using flow cytometry analysis. RESULTS: (1) The serum levels of total IgG and anti-dsDNA antibody in un-treated model group were higher than that in other groups. There was no differences among LR treated group, prednisone treated group and control group. (2) The percentages of CD3+, CD4+, CD8+ T lymphocytes and CD19+, CD23+ B lymphocytes in model group were obviously higher than that in normal control. (3) Compared to un-treated model group, the percentages of CD4+, CD8+ T lymphocytes and CD19+, CD23+ B lymphocytes in LR or prednisone treated group were significantly reduced, which closely reached the levels in normal group. CONCLUSIONS: The immune functions of T and B lymphocytes in BXSB mice are up-regulated. LR inhibits the activation of T and B lymphocytes, reduces the serum levels of IgG and auto-antibody production.     

Elevated levels of serum IL-6, ICAM-1 and P-selectin in stable survivors with liver transplantation

AIM: To study the profile of serum IL-6， ICAM-1 and P-selectin in stable survivors with clinical liver transplantation (LTx). METHODS: Flow cytometric analysis was used to determine the phenotype of T cell subsets in peripheral blood mononuclear cells （PBMCs） from stable survivors with liver transplantation (n=22), and healthy volunteers (n=12). Serum levels of the pro-inflammatory cytokines, tumor necrosis factor (TNF-α) and interleukin (IL-6), intercellular adhesion molecules (ICAM-1) and P-selectin in stable survivors with liver transplantation and healthy volunteers were assessed by enzyme-linked immunoabsordent assay (ELISA). Recently performed 6 cases of liver transplantation were also dynamically observed in this study. RESULTS: Percentage of CD4+ T cells， CD8+ T cells and CD3+ T cells, as well as ratio of CD4 to CD8 were no difference between two groups (P>0.05). However, a significant higher percentage of CD3+CD25+ T cells was found in stable liver transplantation group as compared to healthy group (P<0.05). Significantly increased concentrations of IL-6, ICAM-1 and P-selectin were found in stable liver transplantation group as compared to healthy group (P<0.05). A high TNF-α level was detected in stable liver transplantation group while no significant difference was found as compared to healthy volunteers group (P>0.05). There was not found no regular change of serum cytokines (IL-6, TNF-α) and adhesion molecules (ICAM-1, P-selectin) in 6 liver transplanted patients during post-operation from day 1 to day 30, indicating that was associated with the different status of patients before or after transplantation. CONCLUSIONS: Our data suggesting that increased levels of ICAM-1 and P-selectin, appears to participate in the processing of immunoregulation to transplanted livers, whereas elevated concentrations of IL-6 appear to be involved in the repair of the injury induced by TNF-α in allo-transplanted livers.     

Effect of E-selectin pretreatment on cerebral ischemia-reperfusion injury in rats

AIM:To study the effect of nasal mucosal tolerance to E-selectin on cerebral ischemia-reperfusion injury.METHODS:Two different doses (single and booster) of E-selectin or PBS were dropped into membrana mucosa nasi of rats. The middle cerebral artery occlusion (MCAO) model referring to Zea Longa method with modifications was performed 48 h after the last dose of E-selectin or PBS. After 2 h ischemia and 22 h reperfusion, the numbers of CD3+CD4+T-lymphocyte and CD3+CD8+T lymphocyte subgroup in the blood were examined with flow cytometry. Rats were killed, then part of the animals was used to measure the cerebral infarction volume by TTC staining. mRNA expressions of E-selectin, ICAM-1 and lymphocyte function-associated antigen-1(LFA-1) were determined by RT-PCR and activity of SOD was determined by xanthinoxidanse method in ischemic cortex of the other part of animals. RESULTS:The ratio of the numbers of CD3+CD4+T-lymphocytes and CD3+CD8+T-lymphocytes increased in E-selectin single pretreatment group (P<0.05). Compared to other groups, E-selectin booster pretreatment group showed decreased CD3+CD8+T-lymphocytes (P<0.05), increased ratio of the numbers of CD3+CD4+T-lymphocytes and CD3+CD8+T-lymphocytes (P<0.05), reduced cerebral infarction volume by 40.87% (P<0.05), heightened activity of SOD (P<0.05), lowed E-selectin mRNA and ICAM-1 mRNA expression (P<0.05), and less tendency of LFA-1 mRNA expression.CONCLUSION:E-selectin induces cerebral ischemic tolerance and relieves cerebral ischemia-reperfusion injury. The mechanisms are related to the changes in the ratio of CD4+T-lymphocyte and CD8+T-lymphocyte. The heightened activity of SOD, the lowed mRNA expressions of E-selectin and ICAM-1, as well as the less tendency of LFA-1 mRNA expression are also involved.     

Immunization with mixed peptides derived from glucose-6-phosphate isomerase induces rheumatoid arthritis in DBA/1 mice

AIM: To establish an animal model of rheumatoid arthritis(RA) in DBA/1 mice induced by immunodominant mixed peptides derived from glucose-6-phosphate isomerase(GPI). METHODS: The DBA/1 mice were immunized with emulsified mixed peptide fragments of hGPI325-339+hGPI469-483 or single peptide hGPI325-339 in complete Freund's adjuvant by subcutaneous injection to induce the model of RA. Body weight, ankle joint symptom scores, the pathological change of the ankle joint, the levels of CD4⁺ T cells in the spleen and peripheral blood, the proportion of iNKT cells in the peripheral blood, and the levels of TNF-α and IL-6 in serum were detected to evaluate and analyze the model. RESULTS: The hind paw of the model mice appeared red swelling on the 8th day, and then aggravated gradually to the limbs. The red swelling reached peak on the 14th day, and then relieved gradually. Inflammation response dominated by lymphocytes and monocytes was observed in the ankle joint. The inflammatory effect of mixed peptides was more obvious than that of the single one(P<0.05). Compared with control group and the mice treated with single peptide, the weight gain was slow, the amount of CD4⁺ T cells in the peripheral blood and spleen were increased, the proportion of peripheral iNKT cells in the inflammatory peak was decreased(P<0.05), and the serum level of TNF-α was increased significantly(P<0.05) in the mice treated with mixed peptide fragments. CONCLUSION: The immunological characteristics of RA model induced by mixed GPI peptides in DBA/1 mice is closer to that in RA patients, especially in the immunopathology of iNKT cells. Therefore, this model can be used as a new tool for studying the mechanism and immunological intervention of RA.     

Effect of ulinastatin plus thymosin-α1 therapy on improving immune function in septic patients

AIM: To investigate the effect of ulinastatin plus thymosin-α1 therapy on improving immune function in septic patients. METHODS: 70 patients were divided into two groups. One group was classical treatment group (CT) with regular therapy and another group was classical treatment plus immunotherapy group (CIT) with ulinastatin plus thymosin-α1 for a week.The immune index before and after treatment on day 0, 1, 3 and 7 was observed, including the clinical and survival data. RESULTS: The most common pathogen of sepsis was bacteria, and infection by fungi was in rare. The common locations of bacteria observed were sputum and abdominal drainage. The level of TNF-α was significant lower in CIT group than that in CT group (P<0.05). IL-10 level was significantly higher in CIT group than that in CT group (P<0.05). IgG level was significant lower in CIT group than that in CT group (P<0.05). No significant difference in the levels of IgA, IgM, C3 and C4 between two groups was observed (P>0.05). CD4+T lymphocytes were significant higher in CIT group than those in CT group (P<0.05). From day 7 to day 28, the lymphocytes and level of HLA-DR in CD14+ monocytes were significant higher in CIT group than those in CT group (P<0.05). The time of mechanical ventilation and vasopressors used in CIT group was shorter than those in CT group (P<0.05). But the length of stay and the cost in ICU showed no significant increase between these two groups (P>0.05). During hospitalization, 20 patients died in the CT group and 13 patients died in CIT group (P<0.05). The long-term survival time in CIT group was longer than that in CT group (P<0.05). CONCLUSION: Immunotherapy in septic patients can decrease TNF-α level and increase IL-10 level. Immunotherapy in septic patients can increase IgG level slightly, CD4+T lymphocyte, and HLA-DR in CD14+ monocytes, which improve the immune paralysis in septic patients. Immunotherapy can shorten the time of mechanical ventilation and vasopressors used, but it doesn’t increase the length of stay and the cost.     

Age-related changes in phenotypes of T lymphocytes in human peripheral blood

AIM:To investigate the details of age-related changes of T lymphocytes in order to seek for sensitive biomarker for immunosenesence.METHODS:Heparin anticoagulated venous blood was collected freshly from young (20-35 years) and elderly (50-75 years) volunteers and three or four color immunofluorescence staining was performed. The nucleated cells were acquired and the phenotypes of T lymphocyte subpopulations were analyzed with flow cytometry.RESULTS:There was no significant difference in percentages of pan-T (CD3⁺), helper T (CD4⁺) and cytotoxic (CD8^＋) T subsets between young and elderly, whereas the density of CD3 molecule (MFI) on T cells in elderly group decreased significantly. It was also found that the rates of CD44⁺ and CD62L⁺ T cell subsets in young group did not have statistical difference from elderly. However, the rates of CD95⁺ pan-T, helper T and cytotoxic T subsets of elderly group were all markedly higher than that in young group.CONCLUSIONS:The relative rates of T cell and its subsets displayed no age-related changes while the density of CD3 was down-regulated during aging in these groups investigated. Moreover, the expression percentage of CD95 (Fas) on T cells increased as aging, suggesting that it is a potential biomarker for evaluating immunosenescence.     

Effects of N-acetylcysteine combined with azithromycin on oxidative stress in rats with chronic obstructive pulmonary disease

AIM: To investigate the effects of N-acetylcysteine (NAC) combined with azithromycin (AZI) on oxidative stress in the rats with chronic obstructive pulmonary disease (COPD). METHODS: Male Wistar rats (n=60) were randomly divided into control group, model group, AZI intervention group,NAC intervention group and AZI+NAC group. The COPD model was established by passive smoking and intratracheal instillation of lipopolysaccharide. Each day 30 min prior to smoking, intragastric administration with AZI, NAC or combination of the 2 drugs was given for AZI, NAC, and AZI+NAC groups, respectively. On the 31st day, all rats were killed following lung function test. Cell counts of bronchoalveolar lavage fluid (BALF) were performed, and the contents of interleukin-8 (IL-8), interleukin-17 (IL-17) and tumor necrosis factor alpha (TNF-α) in BALF were measured by ELISA. The histopathology of the lung tissues was observed under light microscope, and the levels of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and malondialdehyde (MDA) in the lung homogenate were measured. RESULTS: Compared with control group, the other 4 groups showed decreased pulmonary function, and inflammatory cell infiltration and alveolar destruction in histopathology. Compared with control group, the other groups showed higher white blood cells, monocyte-macrophages, neutrophils and lymphocytes in the BALF (P<0.05). Compared with model group, AZI group and NAC group, lower white blood cells, neutrophils and lymphocytes in the BALF were observed in AZI+NAC group (P<0.05). Compared with model group, IL-8, IL-17, TNF-α and MDA in AZI group, NAC group and AZI+NAC group significantly decreased (P<0.05), while SOD and GSH-Px significantly increased (P<0.05). Compared with AZI or NAC group, IL-8, IL-17, TNF-α and MDA in AZI+NAC group significantly decreased (P<0.05), while SOD and GSH-Px increased significantly (P<0.05). CONCLUSION: Both NAC and AZI attenuate the lung inflammation and oxidative damage in COPD model rats. Combined medication exerts preferable anti-oxidation effects, which might be more suitable for the treatment of COPD.     

Effects of glucocorticoid on miRNA-155 expression in CD4+T cells of asthmatic mice

AIM:To investigate the effects of glucocorticoid on the regulation of microRNA-155 (miRNA-155) expression in the CD4⁺ T cells of asthmatic mice. METHODS:The ovalbumin (OVA)-induced asthma mouse model was established and the mice were treated with glucocorticoid. The effects of glucocorticoid on the pulmpnary histopathological changes, the expression of miRNA-155 in the lung tissues and CD4⁺T cells, and the levels of cytokines in the bronchoal-veolar lavage fluid (BALF) were evaluated. RESULTS:The results of RT-qPCR showed that the expressions of miRNA-155 in the lung tissues and CD4⁺T cells from the spleen of asthmatic mice were significantly increased, and the level of miRNA-155 in the CD4⁺T cells was significantly increased with the increase in the allergen exposure time (P<0.01). HE and PAS staining showed that OVA significantly increased inflammatory cell infiltration as compared with control group, and the peribronchial and perivascular inflammation and mucus secretion of proliferative goblet cells were significantly reduced after glucocorticoid treatment. Glucocorticoid treatment inhibited the increase in the proportion of CD4⁺ CD8^- cells in the spleen and decreased the accumulation of CD4⁺ T cells in the lung tissues of asthmatic mice (P<0.01). After glucocorticoid treatment, the levels of interleukin-4 (IL-4), IL-5 and IL-13 in BALF were decreased, while the level of interferon-γ was increased significantly (P<0.01). CONCLUSION:Glucocorticoid reduces the accumulation of CD4⁺ T cells and inhibits the expression of miRNA-155 in the lung tissues and spleen CD4⁺ T cells of asthmatic mice.     

Relationship between invasion of tumor-associated macrophages and phenotype and immune efficacy of tumor-infiltrating lymphocytes in advanced ovarian carcinoma

AIM:To explore the relationship between the invasion of tumor-associated macrophages(TAM) and the phenotype and immune efficacy of tumor-infiltrating lymphocytes(TIL) in advanced ovarian carcinoma. METHODS:Immunohistochemical analysis of TAM density in 175 cases of poorly-differentiated ovarian cancer tissue biopsy was performed. The cases were divided into TAM high-density(TAM^High) group and TAM low-density(TAM^Low) group according to the median of TAM density. The control group included 32 cases of benign ovarian lesions. The changes of CD8⁺ and CD25⁺ phenotypes of TIL were detected by flow cytometry analysis. TIL in the 2 groups were cultured in vitro and the conditioned-medium was collected for detecting the expression of IL-2, IL-10, TGF-β and IFN-γ by ELISA. RESULTS:The average TAM infiltration density was 62.8/high-power field(HP, ×400) in 175 cases of poorly-differentiated ovarian carcinoma, and the median was 53.3/HP. TAM^High group was 87 cases and TAM^Low group was 88 cases. A significant difference between malignant ovarian carcinoma group and control group(10.5/HP) was observed. The mean expression of CD8⁺ TIL in TAM^High group was 24%, and CD8⁺ TIL in TAM^Low group was 52%(P<0.05). The mean expression of CD25⁺ TIL in TAM^High was 48%, and CD25⁺ TIL in TAM^Low was 25%(P<0.05). The average infiltration density of CD8⁺ and CD25⁺ TIL in control group was 7%. The average infiltration density of CD8⁺ and CD25⁺ TIL in TAM^High and TAM^Low groups was significantly higher than that in control group(P<0.05). Compared with TAM^Low group, TIL destruction cytokines IL-2 and IFN-γ were significantly decreased in TAM^High group(P<0.05), while the inhibitory cytokines IL-10 and TGF-β were significantly increased(P<0.05). CONCLUSION:In high-density TAM infiltration of ovarian cancer tissues, CD25⁺ TIL type and inhibitory cytokines IL-10 and TGF-β increase, while CD8⁺ TIL type and destruction cytokines IL-2/IFN-γ decrease, suggesting that the high-density TAM has relationship with the phenotype and immune efficacy of TIL.     

Effect of mesalazine on Th1, Th17 and Treg cells in mice with DSS-induced ulcerative colitis

AIM: To investigate the effect of mesalazine treatment on regulation of Th1, Th17 and Treg cells in mice with dextran sulfate sodium (DSS)-induced ulcerative colitis (UC). METHODS: The expression of IL-17, IFN-gamma and Foxp3 in the peripheral blood mononuclear cells (PBMC) and intestinal mucosa lamina propria mononuclear cells (LPMC) of DSS-induced UC mice was detected by flow cytometry analysis. The effect of mesalazine treatment on regulaiton of Th1, Th17 and Treg cells in the mice with DSS-induced ulcerative colitis was examined.RESULTS: The expression of IL-17, IFN-γ and Foxp3 on CD4+T cells were significantly higher in the PBMC of DSS-induced mice than those in control group. CD4+ IFN-γ+T cells and CD4+ Foxp3+T cells were higher in LPMC than those in control group, except CD4+IL-17+T cells. Moreover, the Th1, Th17 and Treg cells were higher in DSS group than those in control group in LPMC. However, only Tregs was higher in PBMC. Pre-treatment with mesalazine significantly decreased the number of Th17, Th1 and Treg cells of UC model mice both in PBMC and LPMC.CONCLUSION: The Th1, Th17 and Tregs cells in DSS-induced mice were significantly higher than those in control mice, suggesting that CD4+T cell subsets play an important role in the pathogenesis of UC. Mesalazine may play a role in the treatment of UC by regulating the Th1, Th17 and Tregs cells.     

Effect of pachyman polysaccharides on Th17/Treg balance in systemic lupus erythematosus patients

AIM: To investigate the immunomodulatory effect of pachyman polysaccharides (PPS) on T helper 17 cell (Th17)/regulatory T cell (Treg) balance in the peripheral blood of systemic lupus erythematosus (SLE) patients. METHODS: The CD4⁺ T cells were isolated from the peripheral blood samples obtained from 45 SLE patients and 35 healthy controls enrolled in our study using magnetic bead separation method. The proportions of Th17 and Treg cells were measured by flow cytometry. The CD4⁺ T cells from SLE patients and healthy controls were treated with PPS. The cytoto-xicity of PPS was evaluated by detecting cell viability with MTT assay. The contents of interleukin-17 (IL-17), IL-6, IL-10 and transforming growth factor-β (TGF-β) were measured by ELISA. The expression of retinoid-related orphan receptor γt (RORγt) and forkhead box protein P3 (Foxp3) at mRNA and protein levels was determined by RT-qPCR and Western blot, respectively. RESULTS: The Th17 cells were significantly elevated, while Treg cells were obviously decreased in the SLE patients compared with the healthy control group (P<0.05). Compare with control group, the contents of IL-17 and IL-6 were decreased, while the contents of IL-10 and TGF-β were increased (P<0.05). The expression of RORγt at mRNA and protein levels was down-regulated and the expression of Foxp3 was up-regulated (P<0.05). The ratio of Th17/Treg was decreased in 100 μg/L nontoxic PPS-treated CD4⁺ T cells isolated from the SLE patients (P<0.05). CONCLUSION: PPS treatment inhibits Th17 cells and elevates Treg cells in the CD4⁺ T cells isolated from SLE patients, which may have a therapeutic effect on SLE patients.     

The influence of anesthesia and means of postoperative pain control on T lymphocyte subtypes of blood in patients with rectal cancer

AIM: To evaluate the influence of anesthesia and different means of postoperative pain control on the T-lymphocyte during the perioperative period in patients with rectal cancer.METHODS: 40 adult patients, aged 65 or older, of American Society of Anesthesiologists (ASA) class 2-3 were divided into two groups according to the type and means of postoperative pain managements. Group Ⅰ (n=20) received intravenous anesthesia and patient controlled analgesia(PCA), fentanyl (13 μg/kg) for post pain; group Ⅱ (n=20) received intravenous anesthesia plus lumber epidural anesthesia and epidural PCA of morphine 5 mg plus ropivacaine 100 mg for post operative pain. Blood samples from internal jugular vein were obtained before surgery, at the completion of surgery and 24, 48, and 120 h post surgery for detecting CD3⁺, CD4⁺, CD4/CD8 counts of peripheral T-lymphocytes. In addition, blood cortisol level and pain intensity were assessed by visual analogue score (VAS)at each time point. RESULTS: Baseline(before anesthesia) values of CD3⁺,CD4⁺, CD4/CD8 in patients were messured and there was a significant decrease of all these values from completion of surgery to 48 h after surgery in both groups (P<0.01). However, group Ⅱ showed a higher CD4⁺ at 48 h, higher CD3⁺,CD4⁺, CD4/CD8 at 120 h post surgery than group Ⅰ (P<0.05). Patients in both groups obtained good pain relief post surgery,but VAS in group Ⅱ were significantly lower than those in group Ⅰ at 24 and 48 h post surgery (P<0.01). Compared with baseline, blood cortisol levels in both groups increased markedly at completion of surgery, and at 24, 48 h after surgery (P<0.01),while the increased cortisol level in group Ⅱ at completion of surgery and 24 h after surgery was less than that in group Ⅰ (P<0.05).CONCLUSION: Combined intravenous anesthesia with lumber epidural anesthesia appears to reduce the perioperative stress response and exerts less negative effects on the T-lymphocytes, suggesting that such a means of anesthesia might be more suitable to the elderly patients with rectal cancer.     

Influence of GPIF on the expression of costimulatory molecules lineaged T cells in mice with immunodeficiency in vivo

AIM:To investigate the effects of goat placenta immunoregulating factor (GPIF) on the expression of costimulatory molecules lineaged T cells in BALB/c mice. METHODS:Animal model for immunodeficiency made from BALB/c mice with whole-body irradiation by 5 Gy 60Coγ-ray was applied for research. The immunosuppressive mice were injected with GPIF for seven days continuously. FACS was applied to analyze the rate of CD28+, CD152+, CD4+CD28+, CD8+CD28+, CD4+CD152+ and CD8+CD152+ cells in splenic lymphocytes and ELISA method was employed to measure the amount of IL-2 and IFN-γ in serum of mice. RESULTS:GPIF increased the percentage of CD28+, CD4+CD28+ and CD8+CD28+ cells (P<0.05, P<0.01), and decreased the percentage of CD152+ (P<0.05, P<0.01), CD4+CD152+ cells (P<0.05, P<0.01) in splenic lymphocytes of immunosuppressive mice significantly. GPIF increased the content of IL-2 and IFN-γ in serum of mice simultaneously (P<0.01). CONCLUSION:Immuno-enhancing effect of GPIF facilitates the costimulation of CD28 pathway, which can activate T cells and accelerate the course of renewing T cell activity. The function of GPIF may have close relationship with an immune network formed by the secretion of IL-2 and IFN-γ and the expression of CD28 and CD152.     

Influence of histone deacetylase inhibitor romidepsin on phenotype and function of T lymphocytes in vitro

AIM: To explore the effects of romidepsin (FK228), a novel histone deacetylase inhibitor, on the effector and regulatory T cells in vitro.METHODS: As the reactive cells, lymphocytes, CD4⁺ T cells and CD8⁺ T cells were labelled with CFSE, and stimulated with anti-CD3 and anti-CD28 mAbs in the presence and absence of different levels of romidepsin (experimental group and positive control group), or PBS (placebo group).After 72 h, the proliferation of the cells was detected in different groups. The lymphocytes were stimulated with anti-CD3 and anti-CD28 mAbs in the presence and absence of different levels of romidepsin (experimental group and positive control group),or PBS (placebo group). After 72 h, the percentage of CD4⁺ Foxp3⁺ T cells and the levels of related cytokines were detected in different groups. RESULTS: The proliferation of CFSE-labelled lymphocytes, CD4⁺ T cells and CD8⁺ T cells triggered by anti-CD3 and anti-CD28 mAbs all were inhibited when cultured with romidepsin at concentrations of 1 μmol/L, 3 μmol/L and 5 μmol/L in a dose-dependent manner (P<0.05). Compared with placebo group, in the presence of anti-CD3 and anti-CD28 mAbs, 1 μmol/L romidepsin did not increase the percentage of CD4⁺ Foxp3⁺ T cells (P>0.05). When cultured with romidepsin at concentrations of 3 μmol/L and 5 μmol/L, the percentage of CD4⁺ Foxp3⁺ T cells was enhanced markedly (P<0.05). The levels of IL-10 and TNF-α in the supernatant were markedly increased in positive control group and 3 experimental groups (P<0.05), and the levels of cytokines in different experimental groups were gradually decreased with the elevation of FK228 concentration (P<0.05). The level of TGF-β was slightly increased in positive control group with no significant difference compared with placebo group (P>0.05). With the increase in the concentration of FK228 in different experimental groups, the TGF-β level was increased in a dose-dependent manner and there were significant differences in the 3 experimental groups. Meanwhile, significant differences existed between experimental groups and placebo group and between experimental groups and positive control group (P<0.05). CONCLUSION: Romidepsin inhibits the proliferation of CD4⁺ and CD8⁺ effector T cells and increases the percentage of CD4⁺ Foxp3⁺ regulatory T cells. It may be related to the increased level of TGF-β, but independent of IL-10.     

Response of human triple-negative breast cancer to paclitaxel after vascular normalization in nude mice

AIM: To explore whether there is synergistic effect of recombinant human endostatin (rh-Endo) and paclitaxel (Pac) in the time window of vascular normalization and the role of magnetic resonance imaging (MRI) in early assessment of chemotherapy by observing the response of human triple-negative breast cancer (TNBC) to Pac after vascular normalization in nude mice. METHODS: The human TNBC MDA-MB-231 cells were planted in the subcutaneous region of right lower abdomen of BALB/c-nu female nude mice. These nude mice were randomly divided into 4 groups (n=7). rh-Endo was given for 17 consecutive days in rh-Endo group and rh-Endo+Pac group. Pac was given on the 6th and 12th days in Pac group and rh-Endo+Pac group. The dosage of both drugs was 10 mg·kg^-1·d^-1 (ip). On the day before the treatment and the 5th, 11th and 17th days after treatment, all the transplanted tumors were examined by MRI. All the mice were killed by cervical dislocation and their transplanted tumors were taken down for examinations after the last MRI on the 17th day. The changes of pathology, immunohistochemisty, microvessel density (MVD) and Ki67 expression were measured. RESULTS: On the 17th day, the volume of transplanted tumor in rh-Endo+Pac group was smaller than that in model group and rh-Endo group (P<0.05), and no difference between rh-Endo+Pac group and Pac group was found. On the 17th day, the tumor inhibitory rates in rh-Endo group, Pac group and rh-Endo+Pac group were 14.61%, 39.08% and 54.79%, respectively. The slow diffusion coefficient in Pac group was increased compared with model group, while it was decreased compared with rh-Endo+Pac group (P<0.05). No distant metastatic lesion in the tumor-bearing mice was observed. The necrotic rates in rh-Endo+Pac group and Pac group were higher than those in model group and rh-Endo group. The MVD in model group was higher than that in the other 3 groups. The MVD in rh-Endo+Pac group was decreased compared with Pac group and rh-Endo group. The Ki67 level in rh-Endo+Pac group was decreased compared with rh-Endo group, and no difference between rh-Endo+Pac group and Pac group was detected.CONCLUSION: In the time window of vascular normalization, the combination of Pac and rh-Endo has a significant antitumor effect on TNBC, but this study did not observe a significant synergistic effect of the 2 drugs. The change of slow diffusion coefficient can predict the therapeutic effect in advance.