Effect of Diallyl Trisulfide on Induction of UDS by Mutagenic Drugs in Primary Rat Hepatocytes

Most of anticancer drugs are mutagenic A possible exception is diallyl trisulfide(DAT).a component of garlic.Its modifying effect on induction of DS by mutagenic mitomycin C(MMC),cyclophosphamide(CP)and cis-diamine dichloroplatin(DDP0was investigated with the UDS assay in the primary cultures of Wistar rat hepatocytes (hpc) using the autoradiorgaphic technique.Results showed that 1.0-4.0 nmol/ml of DAT did not induce UDS and that MMC,CP and DDP resulted in a significant induction of dose-dependent UDS.DAT enhanced induction of UDS by these drugs.A dose-effect relationship was obsverved between dos of DAT and enhancement of induction of UDS.However,the mechanism of the enhancement is not clear.     

Effects of Ischemia and Anoxia on Cell Activation and Cell Cycle of Cultured Astrocytes in vitro

Over the past years,the effects of astrocyteson the stability and plasticity of the internal milieuof the central nerve system(CNS),and on theregulation of the microenvironment and regenera-tion of neurons are increasingly drawing the atten-tion of researchers.Studies showed that astrocytesbecame abnormally active after cerebral ischemia,with a unique temporal and spatial pattern,andpresenting complicated changes at molecular leveland showing a dual effect:adverse and beneficial,on the repair …     

Effect of Antisense Oligonucleotide to Annexin Ⅱ on the t—PA—mediated Plasminogen Activation in vitro

Summary: In order to study the role of annexin Ⅱ , a recombinant expression vector, pZeoSV2 (+)ANN Ⅱ , containing the annexin Ⅱ cDNA, was developed. The 1.1-kb-length annexin ⅡeDNA was inserted into a expression vector, PZeoSV(+) and transfected into HL-60 cells which had low baseline expression of Ann- I. pZeoSV(+) ANN I was analyzed by restriction mapping and the Ann-I sequence identified. The ability of the transfected cells, non-transfected and mock-transfected cells to stimulate t-PA-depend plasminogen activation was compared. The results showed that HL-60 with pZeoSV (+)ANN I transfection could significantly increase the plasminogen activation (8. 9± 1.2 U) in vitro with the difference being significant as compared with non-transfected (1.5±0. 4 U) and mock-transfected cells (4.2±0. 9 U), respectively. Antiannexin Ⅱ oligonucleotides significantly inhibited the binding ability of t-PA and plasminogen to annexin Ⅱ , and obviously reduced the plasminogen activation in vitro. The above findings showed human umbilical vein endothelial cells (HUVECs) treated with sense or missense oligonucleotides indicated no significant change in binding of t-PA and PLG. Treatment of HUVECs with antian nexin Ⅱ oligonucleotides could significantly reduce the plasminogen activation by 2.4± 0. 3 U as compared with sense oligonucleotide group in binding of t-PA and PLG. These results, therefore,suggest that Ann- Ⅱ can bind plasminogen and participate in the stimulation of t-PA-dependent ac tivation of plasminogen, and that interference with Ann- Ⅱ mRNA by antisense oligonucleotidemay be a new strategy for the therapy of bleeding in patients with hyperfibrinolysis.     

Effect of Cigarette Smoke Extract on the Proliferation of Human Airway Epithelial Cells and Expression and Activation of FAK

The effect of cigarette smoke extract (CSE) on the proliferation of human airway epithelial cells and the possible mechanism was studied. After airway epithelial cells were treated with different concentrations of CSE for 24 h, the cell proliferation was measured by MTT and the distribution of different cell cycles by flow cytometry. The FAK expression level was detected by Western blot and the degree of tyrosine phosphorylation by immunoprecipitation. The results showed that CSE could inhibit the proliferation of human airway epithelial cells, arrest the epithelial cells in G1 phase of cell cycle, dramatically decrease the number of epithelial cells in S and G2 phases； Meanwhile CSE could decrease the expression level of FAK and the degree of its tyrosine phosphorylation. The above effects of CSE were concentration-dependent. The expression of FAK and the degree of its phosphorylation was positively correlated to the increased number of epithelial cells in G1 phase, and negatively to the number of epithelial cells in S and G2 phases. It was concluded that the mechanism by which CSE could inhibit the proliferation of human epithelial cells was contributed to the increased expression and activation of FAK.     

Effect of Trichosanthin on Cell Cycle and Apoptosis of Murine Melanoma Cells

Aplantprotein,calledTrichosanthin(TCS),extractedfromtherootsofTrichosantheskirilowiigrowinginChina.Trichosanthinhasattractedtheattentionofmanyinvestigatorswithitsactivityonfouraspects:theinfluenceonimmunesystem,theinductionoflaborandtheinhibitionofHIVreplicationaswellasthesuppressionoftumorgrowth('--').Inordertoestimatetheprospectoftrichosanthinontumortherapy,theeffectoftrichosanthinonmurinemelanomacellswasstudiedandcomparedamongthedifferentcomPOnentsoftrichosanthinextractedbyourlaboratory…     

Effect of Trichosanthin on Cell Cycle and Apoptosis of Murine Melanoma Cells

Effect of Trichosanthin on Cell Cycle and Apoptosis of Murine Melanoma Cells@毕黎琦@李洪军@张玉华     

Expression of PD1 and BTLA on the CD8+ T Cell and γδT Cell Subsets in Peripheral Blood of Non-Small Cell Lung Cancer Patients

Objective To investigate the expression and regulation of programmed cell death protein 1 (PD1), B lymphocyte and T lymphocyte attenuator (BTLA) in peripheral blood of patients with non-small cell lung cancer (NSCLC); to examine the correlation of the mRNA levels between PD and BTLA in NSCLC. Methods Flow cytometry was used to detect the expression of PD1 and BTLA on the surfaces of CD8⁺ T cells and γδ⁺ T cells in the peripheral blood samples collected from 32 in-patients with stage IV NSCLC and 30 healthy individuals. We compared the expression of PD1 and BTLA on the surfaces of γδ⁺ T cells in the NSCLC patients with bone metastasis before and after the treatment of zoledronic acid. The correlations of PD1 and BTLA, as well as their ligands were analyzed using Pearson correlation analysis with the cBioPortal data platform. Results The frequency of PD1 on the surfaces of CD8⁺ T cells was significantly higher than that of the γδT cells in both healthy controls (t=2.324, P=0.024) and NSCLC patients（t=2.498, P=0.015). The frequency of PD1 on CD8⁺ T cells, rather than on γδ⁺ T cells, was significantly upregulated in advanced NSCLC patients compared with that in healthy controls (t=4.829, P<0.001). The PD1⁺ BTLA⁺γδT cells of the healthy controls were significantly lower than that of the NSCLC patients (t=2.422, P=0.0185). No differences in percentage of PD1⁺γδ⁺ and BTLA⁺γδ⁺ T cells were observed in 7 NSCLC patients with bone metastasis before and after zoledronic acid treatment. PD1 was positively correlated with BTLA in both lung adenocarcinoma (r=0.54; P<0.05) and lung squamous cell carcinoma (r=0.78; P<0.05). Conclusions The upregulation of co-inhibitory molecules occurs on the surfaces of both CD8⁺ T cells and γδT cells in advanced NSCLC, suggesting that these molecules were involved in regulating the inactivation of CD8⁺ T cells and γδ⁺ T cells, immune escape and tumor invasion.     

Inhibitory Effect of Matrine on the Expression of PSA and AR in Prostate Cancer Cell line LNCaP

In order to investigate the inhibitory effect of matrine on the expression of prostate specific antigen （PSA） and androgen receptor （AR） in prostate cancer cell line LNCaP in vitro, LNCaP cells were treated with matrine at different concentrations （0.5, 1.0, 1.5, 2.0 g/L） for 12-36 h. The growth activities of cancer cells were determined by MTT colorimetric assay. The AR level was measured by Western blotting. The expression of PSA was detected by using AXSYM system-chemical luciferase methods. The results showed that matrine could effectively inhibit the growth of androgen-dependent prostate cancer cell line LNCaP in vitro in a time-and dose-dependent manner （P〈0.05）. It could obviously decrease the level of AR （P〈0.01） and inhibit the expression of PSA in a dose-dependent manner （P〈0.05） in LNCaP cells. It was concluded that matrine could significantly suppress the growth of LNCaP cells and inhibit the expression of PSA and AR of prostate cancer cells.     

The Effect of Beta—Amyloid on Neurons and the Influence of Glucocorticoid and Age on Such Effect

Alzheimer disease( AD) has been a commoncause that leads to the loss of mental function inthe elderly and the pathogenesis of which remainunclear.The recent studies showed that the de-posit ofβ- amyloid ( Aβ) mightbe a late- stage com-mon pathway shared by such inducing factors ashereditary,environmental factors and age etc.Many researchers have reported that high level ofAβ mightinduce the apoptosis even necrosisof cul-tured hippocampal neurons under different condi-tions.Butthe reports …     

Effect of N-tosyl-L-phenylalanylchloromethyl Ketone on Tumor Necrosis Factor-alpha -induced NF-κB Activation and Apoptosis in U937 Cell Line

Summary: To investigate the effect of N-tosyl-L-phenylalanylchloromethyl ketone (TPCK) on tumor necrosis factor-alpha-induced NF-κB activation and apoptosis in U937 cell line, changes and subcellular localization of NF-κB/p65 and IκB-α were observed by fluorescencemicroscopy and expression and degradation of IκB-α by flow cytometry. The apoptosis of U937 cells was measured by flow cytometry and electrophoresis of DNA. Immunolfluorescence assay showed that NF-κB/p65,IκB-α only localized in cytoplasm. After TNF-α stimulation, p65 was localized only in nuclei, and IκB-α was only localized in cytoplasm and decreased. The changes of TNF-α stimulation were specifically inhibited by TPCK. Flow cytometry also revealed the downregulation of IκB-α protein during TNF-α-induced apoptosis and the down-regulation was specifically inhibited by TPCK. Flow cytometry also showed the apoptosis of U937 cells after TNF-α induction. DNA ladder can be detected in cells treated by TNF-α. It is concluded that degradation of IκB-α protein and NF-κB/p65 translocation occur during TNF-α-induced apoptosis of U937 cells, suggesting the activation of NF-κB.TPCK-sensitive protease plays an important role in the degradation of IκB-α protein induced by TNF-α in U937 cells. TPCK sensitive protease also plays an important role in the apoptosis of U937 cells induced by TNF-α.     

二烯丙基三硫化物涂层支架对冠状动脉损伤后血管壁内iNOS蛋白表达及NO水平的影响

目的:研究二烯丙基三硫化物涂层支架对犬冠状动脉损伤后血管壁内诱导型一氧化氮合酶(iNOS)蛋白表达及NO水平的影响. 方法:进行冠状动脉左回旋支支架置入术的犬,将二烯丙基三硫化物涂层支架和对照支架分别置入左回旋支的远端和近端. 术后1,3,10和28 d分别处死动物,蛋白质印迹杂交法测定血管壁中iNOS蛋白的表达、硝酸还原酶法检测NO水平. 结果:对照支架组术后1 d血管壁中有少量iNOS蛋白表达,3 d表达增加,10 d达高峰,28 d时仍有表达,二烯丙基三硫化物涂层支架组iNOS蛋白在各时间点的表达量均高于对照支架组;对照支架组术后1 d时NO生成量低于正常组织,3 d时NO水平升高,10 d时最高,28 d时降至基线水平,二烯丙基三硫化物涂层支架组各时间点的NO水平明显高于对照支架组. 结论:二烯丙基三硫化物涂层支架可显著增加iNOS表达,并提升NO产量,提示二烯丙基三硫化物涂层支架可能是抑制内膜增生的一个重要机制.     

几种肿瘤免疫抑制因子作用的研究

通过体外细胞培养，采用ＭＴＴ法和ＮＯ测定法，观察了淋巴瘤（Ｒａｊｉ）、实体瘤（Ｓ_（１８０））、艾氏腹水瘤（ＥＡＣ）、肝癌（ＨＣ）４种肿瘤细胞产生的肿瘤免疫抑制因子（ＴＤＳＦ）对小鼠腹腔巨噬细胞产生ＮＯ和Ｔ淋巴细胞激活的影响。结果发现Ｒａｊｉ－ＴＤＳＦ和Ｓ_（１８０）－ＴＤＳＦ对巨噬细胞产生ＮＯ均有明显的促进作用（Ｐ＜０．０１，Ｐ＜０．０５）．而ＥＡＣ－ＴＤＳＦ和ＨＣ－ＴＤＳＦ则否（Ｐ＞０．０５），它们对Ｔ淋巴细胞激活的抑制作用Ｓ_（１８０）－ＴＤＳＦ和ＥＡＣ－ＴＤＳＦ可被大蒜素（ＤＡＴＳ）所拮抗，与无ＤＡＴＳ组比较抑制率明显下降（Ｐ＜０．０１）；而ＤＡＴＳ却不能拮抗Ｒａｊｉ－ＴＤＳＦ和ＨＣ－ＴＤＳＦ的抑制作用（Ｐ＞０．０５）。结果表明，不同肿瘤细胞产生的ＴＤＳＦ的作用及其机制可能是多样的。     

Protective effect of diallyl trisulfide on liver in rats with sepsis and the mechanism

The protective effects of diallyl trisulfide on liver were examined in rats with sepsis. Sepsis was reproduced in rats by cecum ligation and puncture (CLP). Fifty-six male Wistar rats were randomly divided into sham-operated group (group S, n=8), sepsis model group (group C, n=24), diallyl trisulfide (DATS)-treated group (group D, n=24). Animals in groups C and D were further divided into three subgroups according to different observation time points, with 8 rats in each sub-group.Rats in group D and C were intravenously injected with normal saline or DATS respectively at a dose of 20 mg/kg after the establishment of sepsis model. Eight rats in groups C and D were sacrificed at 3, 6 and 24 h post-CLP and their livers were harvested for detection of interleukin (IL)-1 receptor associated kinase-4 (IRAK-4), nuclear factor-κB (NF-κB), c-fos, c-jun, malondialdehydethhe (MDA) and superoxide dismutase (SOD), tumor necrosis factor alpha (TNF-α) and for pathological examination. The results showed that the levels of serum IRAK-4, NF-κB and TNF-α in hepatic tissues were higher in group C than group S (control group) (P<0.05). After DATS treatment, the levels of IRAK-4 and NF-κB in the hepatic tissues and serum TNF-α in group D were lower than those in group C (P<0.05). The levels of c-fos and c-jun and MDA in the hepatic tissues were higher in group C than in group S (P<0.05). After DATS treatment, the levels of c-fos and c-jun and MDA in the hepatic tissues were significantly lower in group D than in group C (P<0.05). When compared with group S group, concentration of SOD in the hepatic tissues in group C was significantly lower (P<0.05). After DATS treatment, the concentration of SOD in the hepatic tissues was higher in group D than in group C (P<0.05). These findings suggested that treatment with DATS could ameliorate sepsis-induced liver injury in rats. The protective effect might be related to its ability to inhibit the signal pathway of IRAK-4 and NF-κB, thereby decreasing the production o     

Investigation on the inhibitory effect of tumor-derived immunosuppressive factor (s) on T lymphocyte proliferation

Using five tumor cell lines, the effect of tumor-derived immunosuppressive factor(s) (TDSF) on T lymphocyte proliferation and its mechanism have been investigated. It was found that TDSF markedly inhibited PHA-stimulated T lymphocyte proliferation via a noncytotoxic mechanism. The inhibition increased in dose-dependent manner and the maximum inhibition was achieved when the factor was added at the initiation of the culture. When PBMC were preincubated with supernatants of tumor cells for 24 h, washed extensively and then cocultured with freshly prepared PBMC, similar suppressive effects were observed. The above results indicated that the activation of any suppressor cell subgroup may be one of the mechanisms of immunosuppressive action of TDSF.     

Effect of immunosuppressive factor produced by human lung cancer cell line A549 on the production and action of interleukin 1

A549, a human lung cancer cell line, spontaneously produces a tumor-derived immunosuppressive factor (TDSF) which inhibits the production and action of interleukin 1 (IL-1). After exposure of macrophages to TDSF for 5 h, the production of IL-1 by macrophages was significantly inhibited. The inhibition was much stronger if TDSF existed continuously in macrophage culture. The response of thymocytes treated with nylon wool to exogenous IL-1 was significantly suppressed in the presence of TDSF, suggesting that TDSF can inhibit the action of IL-1. The thymocytes untreated with nylon wool could proliferate after being stimulated with Con A. The proliferation was significantly suppressed by TDSF in a dose-dependent manner. These results suggest that the inhibitory action of TDSF on T cell activation is associated with IL-1, and that TDSF might exert an inhibitory action on other reactions mediated by IL-1. Furthermore, TDSF can reduce the supplementation of new T cells by inhibiting the proliferation of thymocytes.     

全氟化碳对兔肺泡巨噬细胞功能的影响

目的：通过观察全氟化碳（ＰＦＣ）对兔肺泡巨噬细胞（ＡＭ）功能的影响;探讨急性肺损伤（ＡＬＩ）动物应用部分液体通气（ＰＬＶ）治疗后肺部炎症减轻的可能原因。方法：兔ＡＭ暴露于全氟萘烷（ＦＤＣ）后,观察在有效刺激下产生过氧化氢（Ｈ２Ｏ２）,肿瘤坏死因子（ＴＮＦ－α）及一氧化氮（ＮＯ）能力的变化。结果：ＡＭ暴露于ＦＤＣ后产生Ｈ２Ｏ２,ＴＮＦ－α和ＮＯ的量明显降低。结论：在体外ＦＤＣ能明显降低ＡＭ对有效刺激     

二烯丙基三硫化物涂层支架对冠状动脉损伤后细胞凋亡和凋亡相关基因蛋白Bcl-x1 表达的影响

目的观察二烯丙基三硫化物涂层支架对犬冠状动脉损伤后细胞凋亡和凋亡相关基因蛋白Bcl-x1表达的影响.方法犬进行冠状动脉左回旋支支架植入术,将二烯丙基三硫化物涂层支架和对照支架分别植入左回旋支的远端和近端,术后28d处死,进行细胞凋亡检测、Bcl-x1免疫组化和蛋白质印迹杂交.结果术后28d,二烯丙基三硫化物涂层支架组细胞凋亡率为(9.18±3.41)%高于对照支架组的(3.28±1.74)%,P＜0.01.对照支架组血管壁内Bcl-x1蛋白表达为,12.17±5.02,显著高于二烯丙基三硫化物涂层支架组的5.46±2.50,P＜0.05.蛋白质印迹杂交也显示对照支架组有高水平Bcl-x1蛋白表达(37.52±6.15),明显高于二烯丙基三硫化物涂层支架组(10.48±2.75),P＜0.01.结论二烯丙基三硫化物可逆转血管损伤后内膜中Bcl-x1的高表达,促进细胞凋亡,可能是二烯丙基三硫化物涂层支架抑制内膜增生的机制之一.     

几丁聚糖对小鼠腹腔巨噬细胞一氧化氮生成和iNOS活性的影晌

目的研究水溶性几丁聚糖对小鼠腹腔巨噬细胞一氧化氮(NO)生成和一氧化氮合酶(iNOS)活性的影响,探讨几丁聚糖的免疫调节作用机制。方法采用Griess反应、荧光法分别测定不同剂量(100mg/kg,200mg/kg,300mg/kg)水溶性几丁聚糖作用的小鼠腹腔巨噬细胞的NO生成量、iNOS活性。结果Griess反应结果显示几丁聚糖提高小鼠腹腔巨噬细胞的NO生成量,低、中、高剂量组NO生成量(nmol/L)分别为51.36±12.14,79.25±14.62,86.44±15.27,与对照组比较有显著性差异(P<0.01);荧光法测定结果显示低、中、高剂量组小鼠腹腔巨噬细胞iNOS活性[nmol/(well.min)]分别为4.06±0.15,6.81±0.13,7.16±0.12,与对照组比较P<0.01。结论水溶性几丁聚糖能提高小鼠腹腔巨噬细胞iNOS活性,增加NO合成。提示几丁聚糖的免疫调节作用可能与几丁聚糖刺激巨噬细胞NO生成有关。     

脐血T细胞亚群的观察

测定脐带血和成人血的Ｔ淋巴细胞亚群（ＣＤ３．ＣＤ４．ＣＤ８）结果提示：脐带血的ＣＤ３．ＣＤ４．ＣＤ８较成人低，两者差异有高度显著性（Ｐ＜０．０１）。说明胎儿的免疫力低于成人，但脐血的Ｔ淋巴细胞在胎儿生长发育的过程中起着相当重要的作用     

雌激素与去卵巢小鼠外周血T淋巴细胞亚群及免疫球蛋白关系的研究

目的 通过对去卵巢小鼠补充E2治疗,观察外周血T淋巴细胞亚群及免疫球蛋白水平变化,探讨E2对上述指标的影响.方法 60只健康雌性小鼠随机分为3组,假手术组不给药物,去卵巢小鼠分别接受苯甲酸雌二醇（用药组）及注射油（非用药组）皮下注射治疗,每周2次,共注射25次（12周）,酶联免疫吸附法检测3组血清E2及免疫球蛋白（IgG、IgM、IgA）水平,流式细胞仪检测血CD4＋ T细胞和CD8＋T细胞变化.结果 （1）术后12 周用药组血CD8＋T细胞数（18.16±4.04）比非用药组（25.73±3.63）降低（P＜0.05）;假手术组血CD8＋T细胞数（16.54±3.12）比非用药组（25.73±3.63）降低（P＜0.05）,差异有统计学意义.（2）与非用药组比较,用药组及假手术组CD4＋ T细胞数呈上升趋势,但差异均无统计学意义（均P ＞0.05）.（3）血清IgG、IgM、IgA水平3组比较差异均无统计学意义（均P ＞0.05）.结论 E2可降低去卵巢小鼠外周血CD8＋T细胞水平,参与免疫系统调节.