Expression of tumor necrosis factor related apoptosis inducing ligand receptor in glioblastoma

BACKGROUND： Receptors for tumor necrosis factor related apoptosis inducing ligand （TRAIL） include death receptor 4, death receptor 5, decoy receptor 1, and decoy receptor 2. Activation of death receptor 4 and 5 selectively kills tumor cells. OBJECTIVE： To detect TRAIL receptor expression in glioblastoma by immunohistochemistry and RT-PCR, and to compare this expression to that in normal brain tissue. DESIGN： Observational analysis. SETTING： Department of Pathology, the First Affiliated Hospital of Zhengzhou University; Henan Tumor Pathology Key Laboratory. PARTICIPANTS： Twenty-five patients （17 males and 8 females） who received glioblastoma resection were selected from the Fifth Affiliated Hospital of Zhengzhou University, between September 2003 to June 2004. All glioblastoma samples were diagnosed pathologically. Twenty patients （12 males and 8 females） with craniocerebral injury who received normal brain tissue resection were selected in the same time period. There were no significant differences in sex and age between glioblastoma patients or between craniocerebral injury patients （P 〉 0.05）. All patients and appropriate relatives provided informed consent, and this study was approved by the local research ethics committee. METHODS： Polyclonal antibody against TRAIL receptors and an immunohistochemical kit （batch number： 200502） were purchased from Boster Company, Wuhan. Immunohistochemistry： Expression of death receptor 4, death receptor 5, decoy receptor l, and decoy receptor 2 were observed in both glioblastoma and normal brain tissue. The experiment was performed according to the kit instructions, and positive staining was brown-yellow. Assessment： There were no positive signals （-）; weakly positive signals, positive cells 〈 25% （＋）; weakly positive signals, positive cells 25%-50% （＋＋）; strongly positive signals, positive cells 50%-75% （＋＋＋）; strongly positive signals, positive cells 〉 75% （＋＋＋＋）. Evaluation： Expression levels of TRAIL receptors were estimated in both normal brain tissue and glioblastoma. Expression of decoy receptor 1 and decoy receptor 2 mRNA in glioblastoma were detected by reverse transcription polymerase chain reaction, and expression of decoy receptor in glioblastoma was estimated. MAIN OUTCOME MEASURES： Comparison of death receptor and decoy receptor protein expression between glioblastoma and normal brain tissue; decoy receptor mRNA expression in glioblastoma. RESULTS： Death receptor protein expression was strongly positive （＋＋＋） in glioblastoma, while it was weakly positive （＋, ＋＋） in normal brain tissue. Therefore, expression rate of death receptor protein in the glioblastoma was significantly higher than that in the normal brain tissue （.~ 2 = 18.48, 23.03, P 〈 0.01）. Decoy receptor protein expression in the glioblastoma was significantly lower than that in the normal brain tissue （ x2 = 6.65, 18.76, P 〈 0.01）. The level of decoy receptor mRNA expression in glioblastoma was significantly higher than those of protein expression （ x 2 = 9.82, 10.09, P〈 0.01）. CONCLUSION： High expression of death receptor and low expression of decoy receptor are frequently observed in glioblastoma, suggesting that TRAIL receptor genes show an anti-tumor and expressive response during the initiation and development of the tumor. There are significant differences in decoy receptor expression between normal brain tissue and glioblastoma, suggesting that the restricted expression of decoy receptor in glioblastoma is regulated at the post-transcriptional level.

Antitumor effect of tumor necrosis factor-related apoptosis inducing ligand combined with mevastatin on a human glioma cell line SWO-38

BACKGROUND： Previous studies have reported that statins are less toxic to the human body and have greater antitumor activity; however, few studies have addressed the antitumor effect of statins combined with tumor necrosis factor-related apoptosis inducing ligand （TRAIL）. OBJECTIVE： To explore the effect of TRAIL combined with mevastatin on the proliferation and apoptotic cell death of a human glioma cell line SWO-38, and to study its mechanism of action. DESIGN, TIME AND SETTING： An in vitro control experiment was performed at the Central Laboratory of the Third Hospital Affiliated to Sun Yat-sen University, between January and April 2009. MATERIALS： The human SWO-38 cell line was provided by Cell Research, Department of Animal Experimental Center of Sun Yat-sen University; human recombinant soluble TRAIL by R＆D, USA; and mevastatin by Sigma, USA. METHODS： SWO-38 cells were separately incubated in TRAIL （100, 200, 300, 400, and 500 tJg/L） and mevastatin （5, 10, 20, 30, and 40 pmol/L） for 72 hours. In addition, SWO-38 cells were incubated in TRAIL （300 μg/L）, mevastatin （30 μmol/L）, and a solution containing both TRAIL and mevastatin for 12, 24, 48 and 72 hours. MAIN OUTCOME MEASURES： Cell proliferation was detected using methyl thiazolyl tetrazolium assay; cell apoptosis was observed using Hoechst 33258 staining and fluorescence microscopy and was measured using Annexin V/propidium iodide flow cytometry; TRAIL R1/DR4 and TRAIL R2/DR5 protein expressions levels were measured using indirect immunofluorescence staining combined with flow cytometry in the recombinant soluble TRAIL （rsTRAIL, 300 tJg/L）, mevastatin （30 IJmol/L） and combination groups; TRAIL R1/DR4 and TRAIL R2/DR5 mRNA expression was detected using real-time polymerase chain reaction. RESULTS： rsTRAIL, mevastatin and their combination inhibited tumor proliferation in a time- and dose-dependent manner. The proliferation inhibitory rate and apoptosis rate of human SWO-38 cells in the combined group were significantly greater than the rsTRAIL or mevastatin alone group （P 〈 0.01）. TRAIL R1/DR4 and TRAIL R2/DR5 protein and mRNA expressions were increased in the combination group compared with mevastatin or rsTRAIL alone after 72 hours （P 〈 0.01）. CONCLUSION： Both rsTRAIL and mevastatin inhibit the proliferation and apoptosis of the human glioma cell line SWO-38, while their combination enhances the anti-tumor effect. The mechanism of action possibly correlates to the upregulation of TRAIL R1/DR4 and TRAIL R2/DR5 mRNA expression by mevastatin, thereby enhancing the cell sensitivity to rsTRAIL.

Tumor necrosis factor-related apoptosis-inducing ligand-induced apoptosis in glioma U87 cells

Studies have shown that tumor necrosis factor-related apoptosis-inducing ligand（TRAIL）exhibits strong induction of apoptosis in human glioma cells.It remains unclear whether the mitochondrion pathway,an important apoptosis signaling pathway,is involved in TRAIL-induced glioma cell apoptosis.In the present study,in vitro cultured human glioma U87 cells were treated with human recombinant soluble TRAIL.Apoptosis of glioma U87 cells,mitochondrial transmembrane potential（Δψm）,cytoplasmic cytochrome c concentration and changes in caspase-3,-8 and-9 activity following human recombinant soluble TRAIL treatment were investigated to determine the mechanism of glioma U87 cell apoptosis induced by TRAIL.Additionally,blocking caspase-8resulted in TRAIL-induced mitochondrion pathway activation,suggesting that TRAIL,through activating caspase-8,initiated a series of mitochondrial events and resulted in apoptosis of glioma U87 cells.

TNF related apoptosis-inducing ligand and its receptors in ocular tumors

Most of the ocular tumors have poor prognosis, and they remain a difficult problem in the area of ophthalmology. With the rapid development of molecular biology and immunologic techniques and the deep research on ocular tumor related genes, it becomes possible to diagnose and treat malignant tumors from the molecular level. The tumor necrosis factor related apoptosis-inducing ligand (TRAIL), a member of the tumor necrosis factor (TNF) super family, is a promising candidate, either alone or in combination with established cancer therapies, since it can initiate apoptosis through the activation of their death receptors. The ability of TRAIL to selectively induce apoptosis of transformed, virus-infected or tumor cells but not normal cells promotes the development of TRAIL-based cancer therapy. Here, we will review TRAIL and its receptors' structure, function, mechanism of action and application in ocular tumors therapy.

Brain edema and tumor necrosis factor-like weak inducer of apoptosis in rats with cerebral ischemia

BACKGROUND： Recent studies have demonstrated that tumor necrosis factor-like weak inducer of apoptosis （TWEAK） participates in brain edema. However, it is unclear whether blood-brain barrier （BBB） disruption is associated with TWEAK during the process of brain edema OBJECTIVE： To investigate the effects of TWEAK on BBB permeability in brain edema. DESIGN, TIME AND SETTING： An immunohistochemical observation, randomized, controlled animal experiment was performed at the Laboratory of Neurosurgical Anatomy, Xiangya Medical College, Central South University ＆ Central Laboratory, Third Xiangya Hospital, Central South University between January 2006 and December 2007. MATERIALS： A total of 48 adult Wistar rats were randomly divided into three groups： normal control （n = 8）, sham-operated （n = 8）, and ischemia/reperfusion （n = 32）. Rats from the ischemia/reperfusion group were randomly assigned to four subgroups according to different time points, i.e., 2 hours of ischemia followed by 6 hours （n = 8）, 12 hours （n = 8）, 1 day （n = 8）, or 12 days （n = 8） of reperfusion. METHODS： Focal cerebral ischemia/reperfusion injury was induced by middle cerebral artery occlusion （MCAO） using the suture method in rats from the ischemia/reperfusion group. Thread was introduced at a depth of 17-19 mm. Rats in the sham-operated group were subjected to experimental procedures similar to the ischemia/reperfusion group; however, the introducing depth of thread was 10 mm. The normal control group was not given any intervention. MAIN OUTCOME MEASURES： TWEAK expression was examined by immunohistochemistry; brain water content on the ischemic side was calculated as the ratio of dry to wet tissue weight; BBB permeability was measured by Evans blue extravasation. RESULTS： A total of eight rats died prior to and after surgery and an additional eight rats were randomly entered into the study. Thus 48 rats were included in the final analysis. In the ischemia/reperfusion group, TWEAK-positive cells were present in the ischemic penumbra surrounding the lamellar necrotic region in the fight cerebral hemisphere at 6 hours reperfusion and increased thereafter; by 2 days reperfusion they had reached a peak level, which was significantly higher than the sham-operated and normal control groups （P 〈 0.05）. At 6 hours reperfusion, both brain water content and Evans blue extravasation showed the same tendency for change as TWEAK expression. Pearson correlation analysis results revealed that the degree of TWEAK expression was positively correlated with brain water content （r = 0.892, P 〈 0.05）. CONCLUSION： The present results confirmed that TWEAK was involved in BBB disruption and participated in brain edema following cerebral ischemia.

Induction of apoptosis in osteogenic sarcoma cells by combination of tumor necrosis factor-related apoptosis inducing ligand and chemotherapeutic agents

Background Osteosarcoma is one of the most common primary malignant tumors of bone with poor prognosis. TNF-related apoptosis inducing ligand （TRAIL） is a member of the tumor necrosis factor （TNF） cytokine family. TRAIL induces apoptosis in various tumor cell lines but is not found to be cytotoxic to many normal cell types in vitro. We investigated the cytotoxic activity of TRAIL and chemotherapeutic agents, including methotrexate （MTX）, doxorubicin （DOX） and cisplatin （CDDP）, on established osteosarcoma cell line-OS-732. Methods OS-732 cells were incubated with chemotherapeutic agents MTX,DOX and CDDP at various peak plasma concentrations（PPC）, 0.1PPC,1PPC and 10PPC, alone or with 100 ng/ml of TRAIL for 24 hours or 48 hours. MTT was used to evaluate the cytotoxic activity of different agents on OS-732. The apoptosis proportion was assayed by flow cytometry. Cellular morphologic changes were observed by phase contrast microscope, scan electron microscope, and transmission electron microscope. Results The inhibitory rate was （24.438±3.414）% with TRAIL of 100 ng/ml for 24 hours. The cells were responsive to DOX and CDDP with a dose-effect relationship （P〈0.05）. In OS-732 cells, DOX and CDDP cooperated synergistically with TRAIL when incubated the cells with them for 24 hours （the combined inhibitory rate is （58.360±2.146）% and （54.101±-2.721）%, respectively）. TRAIL alone or drugs alone induced the apoptosis rate was less than 25% （P〈0.05）. However, the combination of TRAIL and MTX did not present synergistic effects on OS-732 cells （P〉0.05, compared with TRAIL alone）. Conclusions Osteosarcoma OS-732 cells were not responsive to TRAIL-induced apoptosis. DOX and CDDP sensitize osteosarcoma OS-732 cells to TRAIL-induced apoptosis. The combination of TRAIL and MTX presented no synergistic effects on killing OS-732 cells.

COMBINATION OF γ-INTERFERON WITH TRAIL AND CISPLATIN OR ETOPOSIDE INDUCES APOPTOSIS IN HUMAN NEUROBLASTOMA CELL LINE SH-SY5Y

Objective To study the effect of γ-interferon （IFNγ）, tumor necrosis factor related apoptosis inducing ligand （TRAIL）, and cisplatin or etoposide induced apoptosis in human neuroblastoma cell line SH-SY5Y and its possible molecular mechanisms. Methods The expressions of Caspase 8 mRNA and protein were detected with RT-PCR and Western blot analysis. The effects of IFNγ, TRAIL, IFNγ ＋ TRAIL, IFNγ ＋ Caspase 8 inhibitor ＋ TRAIL, IFNγ ＋ cisplatin ＋ TRAIL, and IFNγ ＋ etoposide ＋ TRAIL on the growth and apoptosis of SH-SY5Y cells were detected with the methods of MTT and flow cytometry. The relative Caspase 8 activity was measured with colorimetric assay. Results Caspase 8 was undetectable in SH-SY5Y cells but an increased expression of Caspase 8 mRNA and protein was found after treatment with IFNγ. SH-SY5Y ceils themselves were not sensitive to TRAIL, but those expressing Caspase 8 after treatment with IFNγ were. The killing effect of TRAIL on SH-SY5Y cells expressing Caspase 8 was depressed by Caspase 8 inhibitor. Cisplatin and etoposide could enhance the sensitivity of TRAIL on SH-SY5Y cells. The relative Caspase 8 activity of SH-SY5Y cells in IFNγ ＋ TRAIL group was significantly higher than those of control group, IFNγ group, TRAIL group, and inhibitor group （ P 〈 0. 01 ）. There was no significant difference among IFNγ ＋ TRAIL group, IFNγ ＋ cisplatin ＋ TRAIL group, and IFNγ ＋ etoposide ＋ TRAIL group. Conclusions IFNγ could sensitize SH-SY5Y cells to TRAIL-induced apoptosis and this may be realized by the up-regulation of Caspase 8. Cisplatin and etoposide could enhance the killing effect of TRAIL on SH-SY5Y cells.

Apoptosis and Expression of Protein TRAIL in Granulosa Cells of Rats with Polycystic Ovarian Syndrome

The relationship between apoptosis of granulosa cells and follicle development arrest in polycystic ovarian syndrome (PCOS) rats, and the contribution of tumor necrosis factor related apoptosis inducing ligand (TRAIL) in apoptosis of granulosa cells were explored. By using sodium prasterone sulfate rat PCOS model was induced. The apoptosis of granulosa cells in ovaries of rats was observed by TdT-mediated dUTP-biotin nick end-labeling (TUNEL), and the expression of TRAIL protein and mRNA in granulosa cells was detected by using immunhistochemical staining and reverse transcription polymerase chain reaction (RT-PCR) respectively. The apoptotic rate and the expression of protein TRAIL in granulosa cells were significantly higher in antral follicles from the PCOS rats than in those from the control rats (P<0.01, P<0.05). There was no significant difference in apoptotic rate and the expression of TRAIL protein in granulosa cells of preantral follicles between the PCOS rats and the control rats (P>0.05). No apoptosis and the expression of TRAIL protein in granulosa cells of primordial follicles were found in the two groups. The expression of TRAIL mRNA was significantly stronger in granulosa cells from the PCOS rats than in those from the con- trol rats (P<0.01). It was suggested that the apoptotic rate in granulosa cells was significantly higher in antral follicle from the PCOS rats than in those from the control rats. TRAIL played a role in regu- lating the apoptosis of granulosa cells in PCOS rats.

人参皂苷Rg3调控RANKL/RANK/TRAF6信号通路对绝经后骨质疏松症大鼠骨代谢、成骨细胞的影响

目的 分析人参皂苷Rg3调控核因子-κB受体活化体配体(RANKL)/核因子-κB受体活化体(RANK)/肿瘤坏死因子受体相关因子6(TRAF6)信号通路对绝经后骨质疏松症大鼠骨代谢、成骨细胞的影响。方法 选取50只健康雌性SPF级SD大鼠作为实验对象,随机取10只大鼠作为对照组,其余40只大鼠建立绝经后骨质疏松症模型,其中30只大鼠建模成功,将30只大鼠随机分为模型组、人参皂苷Rg3低剂量组、人参皂苷Rg3高剂量组,各10只。观察各组大鼠股骨病理组织,比较各组大鼠骨代谢指标、骨形态学指标、碱性磷酸酶(ALP)、甲状旁腺激素(PTH)、骨钙素、硬骨素水平,以及RANKL/RANK/TRAF6信号通路相关信使RNA(mRNA)及蛋白表达水平。结果 对照组大鼠骨小梁排列正常,成骨细胞、类骨质面积未发生变化,模型组大鼠成骨细胞数量减少。与模型组比较,人参皂苷Rg3低剂量组、人参皂苷Rg3高剂量组成骨细胞数量增多,且人参皂苷Rg3高剂量组成骨细胞数量比人参皂苷Rg3低剂量组多。各组大鼠骨小梁数量(TB.N)、骨小梁厚度(TB.Th)、总Ⅰ型胶原羧基端前肽(PⅠNP)、N-端骨钙素(N-MID)、PTH、骨钙素、硬骨素、RANKL mRNA、RANK mRNA、TRAF6 mRNA、RANKL蛋白、RANK蛋白、TRAF6蛋白水平比较,对照组>人参皂苷Rg3高剂量组>人参皂苷Rg3低剂量组>模型组,任意两组间比较,差异均有统计学意义(P<0.05)。各组大鼠骨小梁分离度(TB.Sp)、ALP水平比较,对照组<人参皂苷Rg3高剂量组<人参皂苷Rg3低剂量组<模型组,任意两组间比较,差异均有统计学意义(P<0.05)。结论 人参皂苷Rg3可改善骨代谢,提高骨钙素、硬骨素水平,使大鼠骨质疏松症得到改善,其作用机制可能与调控RANKL/RANK/TRAF6信号通路有关。

TRAIL-induced apoptosis of hepatocellular carcinoma cells is augmented by targeted therapies

AIM:To analyze the effect of chemotherapeutic drugs and specific kinase inhibitors,in combination with the death receptor ligand tumor necrosis factor-related apoptosis inducing ligand(TRAIL),on overcoming TRAIL resistance in hepatocellular carcinoma(HCC)and to study the efficacy of agonistic TRAIL antibodies,as well as the commitment of antiapoptotic BCL-2 proteins, in TRAIL-induced apoptosis. METHODS:Surface expression of TRAIL receptors (TRAIL-R1-4)and expression levels of the antiapoptotic BCL-2 proteins MCL-1 and BCL-xL were analyzed by flow cytometry and Western blotting,respectively. Knock-down of MCL-1 and BCL-xL was performed by transfecting specific small interfering RNAs.HCC cellswere treated with kinase inhibitors and chemotherapeutic drugs.Apoptosis induction and cell viability were analyzed via flow cytometry and 3-(4,5-Dimethyl-thiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. RESULTS:TRAIL-R1 and-R2 were profoundly expressed on the HCC cell lines Huh7 and Hep-G2. However,treatment of Huh7 and Hep-G2 with TRAIL and agonistic antibodies only induced minor apoptosis rates.Apoptosis resistance towards TRAIL could be considerably reduced by adding the chemotherapeutic drugs 5-fluorouracil and doxorubicin as well as the kinase inhibitors LY294002[inhibition of phosphoinositol- 3-kinase(PI3K)],AG1478(epidermal growth factor receptor kinase),PD98059(MEK1),rapamycin(mam- malian target of rapamycin)and the multi-kinase inhibitor Sorafenib.Furthermore,the antiapoptotic BCL-2 proteins MCL-1 and BCL-xL play a major role in TRAIL resistance:knock-down by RNA interference increased TRAIL-induced apoptosis of HCC cells.Additionally, knock-down of MCL-1 and BCL-xL led to a significant sensitization of HCC cells towards inhibition of both c-Jun N-terminal kinase and PI3K.CONCLUSION:Our data identify the blockage of survival kinases,combination with chemotherapeutic drugs and targeting of antiapoptotic BCL-2 proteins as promising ways to overcome TRAIL resistance in HCC.

EGFR inhibitors sensitize non-small cell lung cancer cells to TRAIL-induced apoptosis

Apoptosis induced by tumor necrosis factor-related apoptosis-inducing ligand(TRAIL) can be regulated by the epidermal growth factor(EGF) signaling pathway.In this study,recombinant adenoviral vectors that encode TRAIL gene from the hTERT/RGD promoter(AdTRAIL) was combined with drugs including gefitinib,elotinib,and cetuximab that inhibit EGFR and the EGF signaling pathway in non-small cell lung cancer(NSCLC) cell lines to investigate their antitumor activity.In vitro,compared to single reagent,AdTRAIL combined with EGFR inhibitors reduced proliferation and enhanced apoptosis in H460,A549,and SW1573 cell lines.Western blot results suggested that these effects were relative to up-regulation of pro-apoptosis protein BAX and down-regulation of p-AKT.In vivo,AdTRAIL combined with cetuximab resulted in a significant growth reduction in H460 xenografts without damage to the main organs of nude mice.Histological examination and TUNEL analyses of xenografts showed that cetuximab enhanced cell apoptosis induced by AdTRAIL.These results indicate that EGFR inhibitors enhanced AdTRAIL anti-tumor activity in NSCLC cell lines and that inhibiting the AKT pathway played an important role in this enhancement.

EGCG Enhances TRAIL-mediated Apoptosis in Human Melanoma A375 Cell Line

Tumor necrosis factor-related apoptosis-inducing ligand （TRAIL） is a promising anti-cancer agent. Epigallocatechin-3-gallate （EGCG） is a polyphenolic constituent of green tea. In this study, inhibitory effect of combined use of EGCG and TRAIL on human melanoma A375 cells was examined and the possible mechanism investigated. The cells were divided into 4 groups： control group, EGCG group （EGCG： 10, 20 μg/mL）, TRAIL group （TRAIL： 25 ng/mL） and EGCG＋TRAIL group （combined group）. The growth inhibition was measured in the A375 cells treated with different concentrations of TRAIL （（25, 50, 75, 100, 125, 150 ng/mL） by MTT assay. The apoptosis was assessed by flow cytometry. The expressions of DR4 and DR5 were detected by flow cytometry and western blotting. The activities of caspase-8 and caspase-3 were determined by colorimetric assay. The results showed that TRAIL could dose-dependently inhibit the growth of A375 cells and the IC50 of TRAIL was 150 ng/mL. The apoptosis rate was 11.8% in the TRAIL group, 5%–7% in the EGCG group and 48.9%–59.1% in the combined group. Significant difference was found in the apoptosis rate between the combined group and the EGCG or TRAIL group （P〈0.05 for each）. The expression of DR4 instead of DR5 was significantly increased in the EGCG group. The activity of caspase-3 rather than caspase-8 was substantially enhanced in the EGCG group. These results suggest that EGCG is useful for the TRAIL-based treatment for melanoma.

Effect of Wenhua Juanbi Recipe(温化蠲痹方) on Expression of Receptor Activator of Nuclear Factor Kappa B Ligand,Osteoprotegerin,and Tumor Necrosis Factor Receptor Superfamily Member 14 in Rats with Collagen-Induced Arthritis

Objective： To study the effect of Wenhua Juanbi Recipe（温化蠲痹方, WJR） on expression of receptor activator of nuclear factor kappa B ligand（RANKL）, osteoprotegerin（OPG）, and tumor necrosis factor receptor superfamily member 14（TNFRSF14, also known as LIGHT） in rats with collagen-induced arthritis（CIA）. Methods： CIA rats were generated by subcutaneous injection of bovine collagen type-Ⅱ at the tail base. Sixty CIA rats were randomly assigned（10 animals/group） to： model, methotrexate（MTX）-treated（0.78 mg/kg body weight）, and WJR-treated（22.9 g/kg） groups. Healthy normal rats（n=10） were used as the normal control. Treatments or saline were administered once daily by oral gavage. Rats were sacrificed at day 28 post-treatment and knee synovium and peripheral blood serum were collected. Toe swelling degree and expression of RANKL, OPG, and LIGHT were determined by Western blot and immunohistochemistry. Results： Compared with the normal group, toe swelling degree was significantly increased in the model group（P〈0.01）. After treatment, toe swelling degree decreased significantly in the WJR and MTX groups compared with the model group（P〈0.01）. Compared with the normal group, expression of RANKL and LIGHT were significantly increased and OPG significantly decreased in peripheral blood and synovium of the model group（P〈0.01）. Conversely, RANKL and LIGHT expression were significantly reduced and OPG increased in the WJR and MTX groups compared with the model group（P〈0.01）. No statistically significant difference existed between WJR and MTX groups. Conclusion： WJR likely acts by reducing RANKL expression and increasing OPG expression, thus inhibiting RANKL/RANK interaction and reducing LIGHT expression, thereby inhibiting osteoclast formation/activation to block bone erosion.

Antitumor and radiosensitization effect of 12C6+heavy-ion irradiation mediated by radiation-inducible gene therapy

Radio genetic therapy which combines gene therapy with radiotherapy has shown promising results in cancer treatment. In this study, an oncolytic adenovirusbased gene therapy system regulated by radiation was constructed to improve the cancer curative effect. This gene therapy system incorporated the radiation-inducible early growth response gene(Egr-1) promoter and the anticancer gene tumor necrosis factor-related apoptosis-inducing ligand(TRAIL). To confirm the antitumor effect of Ad-ET combined with^12C^(6+)tion irradiation, the survival and apoptosis fraction of tumor cells HT1080 and normal cells MRC-5 in combination treatment were detected by CCK-8 assay and FACS analysis. Then the expression levels of TRAIL gene and protein were tested by real-time PCR and western blotting. The results show that^12C^(6+)tion irradiation could induce cell growth inhibition and apoptosis by activating the TRAIL gene expression in tumor cells, while exhibiting no obvious toxicity to the normal lung cell line MRC-5. Theresults also demonstrate that use of an oncolytic adenovirusbased radiation-inducible gene therapy system together with^12C^(6+)tion irradiation could cause synergistic antitumor effect specifically in tumor cells but not in normal cells. The results indicate that the novel radio genetic therapy could potentiate radiation treatment by improving the safety and efficiency of monotherapy, and provide theoretical support for clinical application of combination treatment.

针灸联合口服复方甘草酸苷治疗寻常型银屑病的效果研究

目的探讨针灸联合口服复方甘草酸苷治疗寻常型银屑病的疗效。方法选取2018年1月至2022年1月收治的寻常型银屑病患者120例随机分为单纯口服复方甘草酸苷组和及针灸联合口服复方甘草酸苷组,每组60例。2组患者均给予复方甘草酸苷片2片/次,3次/d。针灸联合口服复方甘草酸苷组加用针灸治疗,血燥型取三阴交、太溪等,血瘀型取血海、膈俞。隔日治疗1次,连续12周。比较2组患者治疗前及治疗后银屑病皮损面积和严重程度指数(PASI)评分,酶联免疫吸附试验(ELISA)测定血清白介素17A(IL-17A)、肿瘤坏死因子α(TNF-α)和肿瘤坏死因子样细胞凋亡弱诱导因子(TWEAK)的水平。另选同期健康体检者60例为健康对照组。结果与健康对照组相比,寻常型银屑病患者血清中的IL-17A、TNF-α和TWEAK水平明显升高(P<0.05),且三者水平有相关性;口服复方甘草酸苷组经治疗12周后,患者PASI评分较前下降,血清中的IL-17A、TNF-α和TWEAK水平较治疗前明显降低(P<0.05);针灸联合口服复方甘草酸苷组经治疗12周后,患者PASI评分较前下降,血清中的IL-17A、TNF-α和TWEAK水平较治疗前明显降低(P<0.05);治疗12周后,针灸联合口服复方甘草酸苷组血清中的IL-17A、TNF-α和TWEAK水平较口服复方甘草酸苷组下降更明显。结论针灸联合口服复方甘草酸苷较单纯口服复方甘草酸苷能更显著地下调血清中IL-17A、TNF-α和TWEAK的水平,改善寻常型银屑病患者的临床症状,值得临床推广。

可溶性肿瘤坏死因子样凋亡弱诱导因子在斑秃病人血清及皮损中的表达及临床意义

目的检测可溶性肿瘤坏死因子样凋亡弱诱导因子(sTWEAK)在斑秃病人血清及皮损中的表达水平,并探讨其在斑秃发病过程中的作用及临床意义。方法以2017年10月至2018年11月在首都医科大学附属北京友谊医院确诊并治疗的斑秃病人70例为斑秃组,依据病情分为轻症组(44例)、重症组(26例)。同期该院因色素痣或皮肤良性肿瘤进行头皮手术的人群65例作为对照组。比较各组sTWEAK及相关细胞因子白细胞介素(IL)-4、IL-12、γ干扰素(IFN-γ)蛋白及mRNA表达水平;采用Pearson法分析两指标间相关性。多元线性回归分析各指标与sTWEAK或脱发严重度工具(SALT)评分的关系。结果斑秃组血清及皮损中sTWEAK表达水平[(196.73±32.48)ng/L,(2.35±0.62)ng/L]高于对照组[(121.75±19.62)ng/L,(1.49±0.45)ng/L](P<0.05)。斑秃组血清及皮损中细胞因子IL-12、IFN-γ表达水平高于对照组(P<0.05),IL-4表达水平比较差异无统计学意义(P>0.05)。斑秃病人血清及皮损中sTWEAK表达水平与IL-12及IFN-γ呈正相关,与IL-4表达水平无明显相关性。轻症组血清及皮损中sTWEAK[(173.25±26.84)ng/L,(2.04±0.57)ng/L]、IL-12及IFN-γ表达水平低于重症组[(236.47±42.02)ng/L,(2.87±0.70)ng/L](P<0.05);轻症组、重症组血清及皮损中IL-4表达水平差异无统计学意义(P>0.05);斑秃病人血清、皮损处sTWEAK、IL-12、IFN-γ与SALT评分均呈正相关(P<0.05)。多元线性回归分析显示,IL-12、IFN-γ与血清和皮损处sTWEAK呈显著正相关;血清及皮损处sTWEAK、IL-12、IFN-γ与SALT评分存在正相关性(P<0.05)。结论斑秃病人血清及皮损中sTWEAK表达水平增加,与IL-12及IFN-γ呈正相关,sTWEAK、IL-12及IFN-γ均与斑秃病情严重程度密切相关,sTWEAK可能参与斑秃炎症反应过程。

血清sTWEAK、Netrin-1联合APACHEⅡ评分对重型颅脑损伤患者术后预后不良的预测价值

目的探讨血清可溶性肿瘤坏死因子样凋亡弱诱导因子(sTWEAK)、神经轴突导向因子-1(Netrin-1)联合急性生理学与慢性健康状况评分系统Ⅱ(APACHEⅡ)评分对重型颅脑损伤患者术后预后不良的预测价值。方法选取2020年6月至2022年6月该院收治的120例重型颅脑损伤患者,根据术后30 d预后情况分为预后良好组和预后不良组。对比两组血清sTWEAK、Netrin-1水平及APACHEⅡ评分。采用单因素和多因素Logistic回归分析重型颅脑损伤患者术后预后不良的影响因素,并据以构建血清sTWEAK、Netrin-1及APACHEⅡ评分联合应用的预测模型,受试者工作特征(ROC)曲线分析血清sTWEAK、Netrin-1水平及APACHEⅡ评分对重型颅脑损伤患者术后预后不良的预测价值。结果预后不良组的重症监护室居住时间长于预后良好组,白蛋白水平、入院时格拉斯哥昏迷评分法评分和血清Netrin-1水平低于预后良好组,多发脑挫裂伤占比、机械通气占比、入院时APACHEⅡ评分和血清sTWEAK、血清肌酐、血尿素氮水平均高于预后良好组,差异有统计学意义(P<0.05)。多因素Logistic回归分析结果显示,多发脑挫裂伤、Netrin-1水平降低、入院时APACHEⅡ评分升高、sTWEAK水平升高为重型颅脑损伤患者术后预后不良的危险因素(P<0.05)。ROC曲线分析结果显示,血清sTWEAK、Netrin-1及APACHEⅡ评分3个指标单独及联合应用时曲线下面积及其95%CI分别为0.742(0.552~0.925)、0.731(0.488~0.963)、0.714(0.502~0.911)、0.882(0.795~0.947)。结论血清sTWEAK、Netrin-1联合APACHEⅡ评分对重型颅脑损伤患者术后预后不良具有较好的预测价值,可为临床治疗方案的制订提供参考。

注意缺陷多动障碍患儿血清ACE2,TWEAK和CCL5水平检测及诊断价值研究

目的 探究血清血管紧张素转换酶2(angiotensin-converting enzyme 2,ACE2),细胞因子肿瘤坏死因子样细胞凋亡弱诱导因子(tumor necrosis factor-like weak inducer of apoptosis,TWEAK)和CC趋化因子配体5(CC motif chemokine ligand 5,CCL5)水平在注意缺陷多动障碍(attention deficit hyperactivity disorder,ADHD)患儿诊断、病情严重程度评估中的价值。方法 选取2022年10月~2023年10月在河北省儿童医院就诊的125例ADHD患儿记为ADHD组,另选105例在河北省妇幼保健中心体检的健康儿童为对照组,根据临床总体印象严重程度量表(CGI-S)将患儿分为轻中度组(n=83)和重度组(n=42)。ELISA方法检测血清ACE2,TWEAK,CCL5,促黄体生成素(LH)和催乳素(PRL)水平,联合型瑞文测验(CRT)和Conners父母症状问卷(PSQ)对患儿认知和行为状况进行评分。Spearman相关性分析重度组血清ACE2,TWEAK,CCL5与CGI-S,CRT,PSQ评分的相关性;受试者工作特征(ROC)曲线分析ACE2,TWEAK,CCL5对ADHD及严重程度的诊断价值。结果 ADHD患儿血清ACE2(284.35±92.34 pg/ml),TWEAK(2.56±0.76 pg/ml)水平低于对照组(379.23±106.28 pg/ml,3.52±1.12 pg/ml),CCL5水平(7.36±2.37ng/ml)高于对照组(5.24±1.63 ng/ml),差异具有统计学意义(t=7.244,7.703,7.753,均P<0.05);重度组患儿CRT,血清ACE2,TWEAK水平低于轻中度组(t=5.318,6.686,6.490),而PSQ,CCL5水平较高于轻中度组(t=5.220,6.134),差异具有统计学意义(均P<0.05);Spearman相关性分析结果显示,重度组患儿血清ACE2,TWEAK水平与CGI-S,PSQ评分负相关(r=-0.432,-0.453;-0.421,-0.426,均P<0.001),与CRT评分呈正相关(r=0.427,0.418,均P<0.001);CCL5与CGI-S,PSQ评分呈正相关(r=0.421,0.433,均P<0.001),与CRT评分呈负相关(r=-0.446,P<0.001)。血清ACE2,TWEAK和CCL5诊断ADHD发生的AUC分别为0.814,0.803和0.807,三者联合诊断的AUC为0.945,优于各自单独诊断(Z=5.439,4.258,5.576,均P<0.001);血清ACE2,TWEAK,CCL5诊断重度ADHD的AUC分别为0.853,0.796和0.805,三者联合诊断的AUC为0.930,优于各自单独诊断(Z=2.604,3.851,3.567,均P<0.001)。结论 血清ACE2,TWEAK在ADHD患儿血清中低表达,而CCL5高表达,三者表达水平具有相关性,并且诊断ADHD发生和严重程度具有较高的价值。

自发性急性脑出血患者血浆sCD163/sTWEAK比值与预后的关系

目的探究自发性急性脑出血(ACH)患者血浆可溶性CD163(sCD163)/可溶性肿瘤坏死因子样凋亡弱诱导因子(sTWEAK)比值与预后的关系。方法纳入ACH患者90例作为病例组,根据格拉斯哥预后评分将病例组分为预后不良组(38例)和预后良好组(52例);另选取同期体检健康者45例为对照组。酶联免疫吸附试验检测血浆sCD163、sTWEAK水平并计算sCD163/sTWEAK比值。分析血浆sCD163、sTWEAK水平及sCD163/sTWEAK比值与临床资料的相关性;Logistic回归分析ACH患者预后不良的影响因素;受试者工作特征(ROC)曲线分析sCD163/sTWEAK比值对ACH患者预后不良的预测价值。结果病例组血浆sCD163、sTWEAK水平及sCD163/sTWEAK比值均显著高于对照组;预后良好组上述指标均低于预后不良组(P<0.05)。预后良好组血肿体积、美国国立卫生研究院卒中量表(NIHSS)评分、高血压及幕下出血比例均低于预后不良组,低密度脂蛋白胆固醇(LDL-C)高于预后不良组(P<0.05)。相关性分析表明,血浆sCD163、sTWEAK水平及sCD163/sTWEAK比值与出血部位、血肿体积、NIHSS评分、白细胞计数、血小板计数、中性粒细胞/淋巴细胞比值(NLR)呈正相关(P<0.05)。Logistic回归分析显示,sCD163/sTWEAK比值、出血部位、血肿体积、NIHSS评分为ACH患者预后不良的影响因素(P<0.05)。ROC曲线结果表明,sCD163/sTWEAK比值评估ACH患者预后不良的AUC为0.850,敏感度和特异度分别为86.84%和69.23%。结论sCD163/sTWEAK比值在ACH患者血浆中水平较高,并与预后不良有关,该值对此类患者的预后有一定预测价值。

血清sTWEAK、sLOX-1与H型高血压合并HFpEF患者颈动脉粥样硬化的关系

目的研究血清可溶性肿瘤坏死因子样凋亡弱诱导因子(sTWEAK)、可溶性凝集素样氧化型低密度脂蛋白受体-1(sLOX-1)与H型高血压合并射血分数保留的心力衰竭(HFpEF)患者颈动脉粥样硬化的关系。方法选取2021年2月至2023年3月沧州中西医结合医院收治的161例H型高血压合并HFpEF患者,按是否发生颈动脉粥样硬化分为硬化组与非硬化组。同期选取80例于我院体检的健康的志愿者作为对照组。收集受试者的临床资料,采用酶联免疫吸附法检测血清sTWEAK、sLOX-1水平,比较H型高血压合并HFpEF患者与对照组血清sTWEAK、sLOX-1水平,采用多因素logistic回归分析患者发生颈动脉粥样硬化的影响因素,受试者工作特征(ROC)曲线分析血清sTWEAK、sLOX-1对患者发生颈动脉粥样硬化的预测价值。结果161例患者中65例发生颈动脉粥样硬化,发生率为40.37%(65/161),未发生颈动脉粥样硬化者96例。两组在年龄、吸烟、血脂四项等方面比较,差异有统计学意义(P<0.05)。硬化组与非硬化组血清sTWEAK水平低于对照组,sLOX-1水平高于对照组(P<0.05);且硬化组sTWEAK水平低于非硬化组,sLOX-1水平高于非硬化组(P<0.05)。多因素logistic回归分析显示,HDL-C、sTWEAK、年龄、LDL-C、sLOX-1水平是患者发生颈动脉粥样硬化的影响因素(P<0.05)。ROC曲线显示,血清sTWEAK、sLOX-1联合检测对发生颈动脉粥样硬化的预测曲线下面积(AUC)为0.842,预测效能高于两指标单独检测。结论H型高血压合并HFpEF患者的血清sTWEAK水平下降、sLOX-1水平上升,与颈动脉粥样硬化发生有关,两者联合检测对颈动脉粥样硬化的发生具有一定预测价值。