Effects of increased human tumor necrosis factor-like molecule 1A expression in peripheral blood of children with acute Guillain-Barre syndrome on interferon-gamma secretion

BACKGROUND： Human tumor necrosis factor-like molecule 1A （hTL1A） is a strong T helper cell type 1 （Thl） co-stimulator. Guillain-Barre syndrome （GBS） is an autoimmune disorder of the nervous system, which is mediated by Thl cells. OBJECTIVE： To determine hTL1A expression in peripheral blood T lymphocytes of acute GBS children and the effects of hTL1A on secretion of interferon-γ. DESIGN, TIME AND SETTING： A randomized, controlled, neuroimmunological in vitro study was performed at the Central Laboratory of First Hospital of Jilin University, China from November 2005 to November 2007. MATERIALS： Venous blood samples were obtained from 6 healthy donors, aged 6-12 years （all routine blood examination items were normal）, and 6 additional children with acute GBS, aged 6-12 years. The GBS children fell ill within 1 week and were not treated with hormones or immunoglobulin Purified recombinant human soluble tumor necrosis factor-like molecule 1A （rhsTL1A, 1 mg/mL, relative molecular mass 22 000, 6× His tag, soluble form） was supplied by the Central Laboratory of First Hospital of Jilin University, China. METHODS： Peripheral blood mononuclear cells were isolated from healthy donors using the standard Ficoll gradient centrifugation and were incubated in 96-well culture plates. The cells were assigned to the following groups： control （2 μg/mL phytohemagglutinin）, 2μg/mL phytohemagglutinin ＋ 25, 100 and 400 ng/mL rhsTL1A. T cell proliferation was quantified using the tritiated thymidine （3H-TdR） method. Serum interferon-γ levels in acute GBS children were detected by enzyme-linked immunosorbent assay （ELISA）. The ratio of hTL1A-positive T cells to CD3-positive T cells in peripheral blood of acute GBS children was determined using flow cytometry. Following in vitro pre-activation of peripheral blood mononuclear cells by 2 μg/mL phytohemagglutinin, the peripheral blood mononuclear cells were treated with 400 ng/mL exogenous rhsTLIA. Finally, peripheral blood mononuclear cell-secreted interferon-γlevels were measured by ELISA. MAIN OUTCOME MEASURES： The following parameters were measured： rhsTLIA stimulation index to stimulate proliferation of T cells; the serum interferon-γ levels in acute GBS children; the ratio of hTL1A-positive cells to CD3-positive cells; the levels of interferon-γ secreted by peripheral blood mononuclear cells in acute GBS children, as well as rhsTL1A-stimulated interferon-γ levels. RESULTS： T cell proliferation assay revealed that the stimulation index in each rhsTL1A group was greater than the control group. The stimulation index of the 400 ng/mL rhsTL1A group was the greatest. Serum interferon-γ levels in acute GBS children were significantly greater than the control group （P 〈 0.05）. The ratio of hTLIA＋ CD3＋ T cells to CD3＋ T cells in acute GBS children was significantly greater than the control group （P 〈 0.01 ）. Phytohemagglutinin stimulated peripheral blood mononuclear cells to a greater extent than 400 ng/mL rhsTL1A in the acute GBS group, and the secreted interferon-γ levels were significantly increased （P 〈 0.05）. CONCLUSION： In T cells pre-activated with 2 μg/mL phytohemagglutinin, proliferation was effectively increased with 400 ng/mL rhsTL1A treatment. Expression of hTLIA was increased in activated T cells from peripheral blood of acute GBS children, followed by increased interferon-γ secretion. These mechanisms are considered to be part of the pathological process that induces the secretion of inflammatory cytokines in GBS syndrome.

ANTI-CANCER EFFECT OF PSP PURIFIED PRODUCTS ON HUMAN TUMOR CELL LINES IN VITRO

The anti-cancer effect of PSP purified products, PSP-A, PSP-B, PSP-C and crude product PSP-Cr was compared on four human tumor cell lines in vitro. It was found that the inhibition rate of cell proliferation of PSP-A was higher than that of PSP-Cr (P<0. 05). On SPC cells, the inhibition rate of PSP-A at a dosage of 1000μg/ml was 62. 7%, being the highest as compared with those on the other three cell lines. Morphological changes were seen in all the four cell lines, especially in SPC cells after PSP-A treatment.

SYNTHESIS AND EXPRESSION OF A GENE FOR HUMAN TUMOR NECROSIS FACTOR-ALPHA (TNF-α)

TNF-α was found originally In sera of Bacillus Calmette Guerln infected mice as a macrophage derived factor. It Is cytotoxlc for tumor cell and less or not toxic to normal cells in vitor. The gene for human TNF-α with E. coli-preferred codons has been designed according to the amino acid sequence deduced from the cDNA. The gene with 504 bp was divided into 27 oligonucleotide fragments having 30. to 40 nucleotides each. The solid phase phosphotriester method was used for the synthesis of these oligonucleotides. The 27 fragments were annealed to three segments and then linked by T4 DNA ligase. The entire gene was incorporated into plasmld PDR540 with Tac promoter which was used to transform E. coli 7118. The expressed protein was estimated by SDSPAGE with a molecular weight of 1. 7×104Da. The cytotoxlc activity of the product against L-929 cell was 1. 0×107units/ml culture.

Controlling malignant pericardial effusion by intrapericardial administration of recombinant mutant human tumor necrosis factor in patients with carcinoma

Objective: To evaluate the therapeutic efficacy of injecting recombinant mutant human tumor necrosis factor (rmhTNF) into pericardial cavity of carcinoma patients with malignant pericardial effusion. Methods: In 20 cases of malignant pericardial effusion, the intrapericardial catheter was inserted into pericardial cavity, and then rmhTNF of 1.5 × 107 U was infused. The infusion was repeated every 5-7 days with the total 4-6 times. If the effusion disappeared, rmhTNF was then used 2 more times and then the intrapericardial catheter was pulled out. Results: Of 20 patients, 14 were complete response (CR), 4 were partial response (PR) and 2 no change (NC). The disappearance of effusion in 6 cases lasted for more than 6 months. Conclusion: Injecting rmhTNF into pericardial cavity may be a better way to control malignant pericardial effusion and has mild side effects.

<i>In Vitro</i>Evaluation System of Pharmacokinetics and Irradiation Effect in Boron Neutron Capture Therapy (BNCT) Using Three-Dimensional Artificial Human Tumor Tissue Model

Boron neutron capture therapy (BNCT) is based on the incorporation of boron-containing drugs to cancer cells and the nuclear reaction of 10B atoms by thermal neutron irradiation results in tumor degeneration. For the development of this therapy, currently, long time and high cost consuming experiments using many animals are required. In this study, we constructed a new in vitro evaluation system for BNCT by combination of an artificial tumor tissue model, comprised of normal human dermal-derived fibroblast (NHDF) and human pancreatic cancer cell line BxPC3, and the optical plastic material CR-39 as a solid state nuclear track detector. Administration of boronophenylalanine (10BPA) as a boron-containing drug and neutron irradiation up to 2.52 × 1012 n/cm2 to the control tissue constructed by NHDF (NHDF3D) and BxPC3 cell loaded tissue (NHDF3D/BxPC3) resulted in detection of 1.6 times higher number of α-ray/recoiled Li particle tracks in NHDF3D/BxPC3 in comparison to NHDF3D, demonstrating that putative irradiation damage to cancer cells can be evaluated by this system. On a cellular level, the hit number of α-ray/recoiled Li particle tracks per single BxPC3 cells and NHDF was evaluated as 5.46 and 1.71, respectively. The tumor and normal tissue ratio (T/N ratio) was 3.19, which was corresponded with those of BPA as 2 - 4 that reported in the previous studies. This new in vitro evaluation system may provide a useful tool for a low cost, labor-saving, and non-animal method for the development of new boron-containing drugs or improvement of BNCT conditions.

Radiation Sensitivity of <i>in Vitro</i>Evaluation System of Pharmacokinetics in Boron Neutron Capture Therapy (BNCT) Using Three-Dimensional Artificial Human Tumor Tissue Model

One of the important matters that must be determined in advance when performing BNCT treatment is the optimization of neutron irradiation time and dose. In this article, following the previous article (2.52 × 1012 n/cm2) (Case 1), double irradiation (5.04 × 1012 n/cm2) was further performed (Case 2) by verifying the radiation sensitivity performance of the artificial tumor tissue NHDF3D/BxPC3 and the possibility of evaluating the optimum neutron dose required for treatment was examined. As a result, although the radiation damage rate in the normal tissue NHDF3D and the tumor tissue BxPC3 increased in proportion to the irradiation dose due to heavy irradiation in Case 1 or more, the increase in the damage rate in the normal tissue exceeded the tumor tissue. Furthermore, the tumor/normal tissue damage ratio T/N ratio showed the maximum value in Case 1, and the dose ratio in Case 2 with a higher dose showed a tendency to decrease. From the above experimental facts, it was shown that irradiation dose optimization is possible to some extent by an evaluation method using an artificial tumor tissue.

Ornithine decarboxylase, mitogen-activated protein kinase and matrix metalloproteinase-2 expressions in human colon tumors

AIM: To investigate the expressions of omithine decarboxylase (ODC), MMP-2, and Erk, and their relationship in human colon tumors.METHODS: ODC activity, MMP-2 expression, and mitogenactivated protein (MAP) kinase activity (Erk phosphorylation) were determined in 58 surgically removed human colon tumors and their adjacent normal tissues, using [1-14C]-ornithine as a substrate, ELISA assay, and Western blotting, respectively.RESULTS: ODC activity, MMP-2 expression, and Erk phosphorylation were significantly elevated in colon tumors, compared to those in adjacent normal tissues. A significant correlation was observed between ODC activities and MMP-2 levels.CONCLUSION: This is the first report showing a significant correlation between ODC activities and MMP-2 levels in human colon tumors. As MMP-2 is involved in cancer invasion and metastasis, and colon cancer overexpresses ODC, suppression of ODC expression may be a rational approach to treat colon cancer which overexpresses ODC.

Synergistic effects of focus ultrasound with different frequency and hematoporphyrin on human tumor cells

Human hematopoietic cell K 562 , human melenoma cell LiBr and human stomach cancer cells were exposed to ultrasound (US, 1.75 W/cm\+2, 1.4, 2.16 and 2.4 MHz) in vitro in the presence or absence of hematoporphyrin (Hp, 100 μg/mL). The cell damaging effects of treatments were determined by means of the Trypan Blue dye exclusion test, MTT test and FDA test. The experimental results showed that the same cell line had different sensibilities to the US of different frequencies, and different cell line had different damage at the same acoustical radiation. The combined treatment with US and Hp enhanced greatly the cell damage, and no sensibility of insonation cells to US with Hp was observed. The cell damage tests showed that the results of MTT test corresponded well with that of Trypan Blue dye test.

Construction of shuttle, expression vector of human tumor necrosis factor alpha (hTNF-α) gene and its expression in a cyanobacterium, Anabaena sp. PCC 7120

The construction of the shuttle, expression vector of human tumor necrosis factor alpha (hTNF-a) gene and its expression in a cyanobacterium Anabaena sp. PCC 7120 was reported. The 700-bp hTNF cDNA fragments have been recovered from plasmid pRL-rhTNF, then inserted downstream of the promoter PpsbA in the plasmid pRL439. The resultant intermediary plasmid pRL-TC has further been combined with the shuttle vector pDC-8 to get the shuttle, expression vector pDC-TNF. The expression of the rhTNF gene in Escherichia coli has been analyzed by SDS-PAGE and thin-layer scanning, and the results show that the expressed TNF protein with these two vectors is 16.9 percent (pRL-TC) and 15.0 percent (pDC-TNF) of the total proteins in the cells, respectively, while the expression level of TNF gene in plasmid pRL-rhTNF is only 11.8 percent. Combined with the participation of the conjugal and helper plasmids, pDC-TNF has been introduced into Anabaena sp PCC 7120 by triparental conjugative transfer, and the stable transgenic strains have been obtained. The existence of the introduced plasmid pDC-TNF in recom-binant cyanobacterial cells has been demonstrated by the results of the agarose electrophoresis with the extracted plasmid samples and Southern blotting with α-32P labeled hTNF cDNA probes, while the expression of the hTNF gene in Anabaena sp. PCC 7120 has been confirmed by the results of Western blotting with extracted protein samples and human TNF-alpha monoclonal antibodies. The cytotoxicity assays using the mouse cancer cell line L929 proved the cyto-toxicity of the TNF in the crude extracts from the transgenic cyanobacterium Anabaena sp. PCC 7120.

Progress in the Research on the Relationship between Infection with Human Papillomavirus and Tumorigenesis

Human papillomavirus(HPV) is a DNA virus that infects the skin and mucous membranes of the human body. Approximately 80% of sexually active women are likely infected with HPV. Cervical cancer is one of the most common malignant tumors and is second in incidence only to breast cancer. Infection with high-risk HPV types is the main risk factor for cervical cancer, which is currently the only malignant tumor with a clearly defined etiology. HPV infection is also closely related to the incidence and development of other malignant tumors. In addition to cervical cancer, HPV can cause other urogenital tumors, as well as tumors in the digestive tract, lungs, eyes, skin, and other organ systems. This paper provides a review of the progress in HPV infection-related research and provides novel ideas for the study of tumor etiology and mechanisms.

Anti-tumor Effects of a Recombinant Fowlpox Virus Expressing Apoptin on Human Cervical Carcinoma in Vivo and in Vitro

Apoptin is a chicken anemia virus-derived,p53-independent,bcl-2-insensitive apoptotic protein with the ability to specifically induce apoptosis of various human tumor cells,but not of normal diploid cells.To explore the application of apoptin in tumor gene therapy,we used a recombinant fowlpox virus expressing apoptin protein （vFV-Apoptin） to investigate the anti-tumor effectes of vFV-Apoptin on human cervical carcinoma（HeLa） cells in vivo and in vitro through 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide（MTT） assay,acridine orage/ethidium bromide（AO/EB） and annexin V staining test,respectively.The results show that vFV-Apoptin inhibites the proliferation of HeLa cells in vitro through inducing the apoptosis of HeLa cells,and the inhibition effect of vFV-Apoptin has a dose-effect and time-effect relationship.The results of animal models show that vFV-Apoptin significantly inhibits tumor growth,extends the lifespan of animals and improves the mean survival.Experimental results indicate that vFV-Apoptin has a potential application in the tumor gene therapy.

Construction of novel tumor necrosis factor-alpha mutants with reduced toxicity and higher cytotoxicity on human tumor cells

Two tumor necrosis factor-a mutants MT1 (32Trp157Phe) and MT2 (2Lys30Ser- 32Trp157Phe) were constructed by site-directed mutagenesis. These mutants were soluble and over-expressed in E. coli. The purity of purified mutants was above 95% by serial chromatography. The results of Western blot indicated that these mutants could be cross-reactive with monoclonal antibody against native hTNF-a. Compared to parent hTNF-a, the cytotoxicity of these mutants on murine fibrosarcoma L929 cell lines reduced 4—5 orders of magnitude but was equivalent to that of native hTNF-a on human tumor cell lines. The LD50 of mutant MT1 was reduced to 0.34% of wild type and the dose of MT2 that resulted in 30% death of mice reduced to less than 1/700 that of parent hTNF-a.

ISOLATION,CHARACTERIZATION AND EXPRESSION OF THE COMPLEMENTARY DNA FOR HUMAN TUMOR NECROSIS FACTOR (TNF-α)

A cDNA for human TNF-α (615bp) was isolated by means of polymerase chain reaction (PCR) using first strand cDNA from PMA-induced HL-60 cells as template. The result from sequencing the 615 bp cDNA fragment indicated that it corresponded to the entire sequence of mature human TNF coding region. Direct expression of mature human TNF was achieved using a plasmid pHT-1 constructed by ligation of the cDNA and a synthetic DNA. The IPTG-induced bacterial product (hTNF) showed cytotoxicity to mouse L-929 cells. The TNF activity was further identified by neutralization of a specific monoclonal antibody against human TNF-α. Approximately 80,000 units of activity were detected per ml of culture at A600=2.

STUDY ON THE RFLPs AND AMPLICATION AND REARRANGEMENT OF THE TRANSFORMING GENES IN PRIMARY HEPATIC CARCINOMA, GASTRIC CARCINOMA AND BRAIN TUMOR WITH SIX HUMAN ONCOGENE PROBES

By using c-Ha-ras-1, N-ras Wigler (left sequence) and P52C.(right sequence), c-sis, v-erbB, c-myc and v-fos oncogenes as probes, restriction fragment length polymorphisms (RFLPs) of tumor tissue DNAs of 95 patients with gastric carcinoma, primary hepatic carcinoma and brain tumor, and those of 90 normal individuals were studied with the techniques of Southern blot and dot blot. Gene amplification and recombination were also examined in some tumors simultaneously. Some alleles of oncogene are reported in Chinese population for the first time. Moreover, the characteristic frequency of some "rare" alleles and genotypes occurred in some tumor samples is significantly higher than that occured in normal individuals. Pedigree analysis for 2 patients showed that some "rare" alleles are also abandant. Besides, gene amplification and recombination were found in some tumors.

肿瘤潜在侯选靶标及标志物ADAM17的生物信息学分析

目的 采用生物信息学分析肿瘤组织潜在侯选靶标及标志物ADAM17的结构和功能特点。方法 运用ProtParam和ProtScale、STRING12.0、The Human Protein Atlas和cBioPortal等不同数据库,对人ADAM17理化性质、跨膜区域、信号肽、核定位序列、磷酸化和糖基化位点、亚细胞定位、二级/三级结构、蛋白互作网络、不同肿瘤组织中的表达以及遗传改变等进行分析。采用免疫组化对肺、结肠组织的肿瘤组织和正常样本ADAM17表达进行验证。结果 人ADAM17是由824个氨基酸构成的亲水性蛋白;分子式为C_(4066)H_(6356)N_(1124)O_(1277)S_(50),理论等电点为5.50,平均亲水性为-0.573;定位于细胞质,具有1个跨膜结构域、1个信号肽以及2个核定位序列;二级结构主要由不规则卷曲组成,存在89个磷酸化位点、5个N-糖基化位点和9个O-糖基化位点;广泛分布于正常组织中,并在膀胱尿路上皮癌、宫颈鳞状细胞癌、结肠癌(COAD)、肾透明细胞癌、肝脏肝细胞癌和肺鳞状细胞癌(LUSC)中高表达,能与金属蛋白酶抑制因子3、表皮生长因子等主要蛋白发生相互作用,参与肿瘤坏死因子、核因子-κB等信号通路。免疫组化验证结果显示,ADAM17在人肺、结肠正常组织中低表达,而在LUSC、COAD组织中高表达。结论 ADAM17可能是肿瘤的潜在候选靶标及标志物。

重组人Ⅱ型肿瘤坏死因子受体-抗体融合蛋白治疗中毒性表皮坏死松解症的疗效及安全性

目的探讨重组人Ⅱ型肿瘤坏死因子受体-抗体融合蛋白(rhTNFR:Fc)治疗中毒性表皮坏死松解症(TEN)患者的临床疗效及安全性。方法回顾性选取2020年1月至2022年1月海南医学院第一附属医院TEN患者20例,均给予rhTNFR:Fc治疗。治疗21 d后,记录TEN患者临床疗效,比较治疗前与治疗后不同时段(治疗后7、14、21 d)的药疹面积和严重程度指数(DASI)评分[DASI评分平均值,50%DASI(DASI50)、75%DASI(DASI75)、90%DASI(DASI90)所占比例]、血清肿瘤坏死因子α(TNF-α)水平、体温下降时间、皮疹控制时间、住院时间及药物治疗的安全性。结果治疗21 d后,TEN患者中,显效18例(90.00%),有效2例(10.00%)。TEN患者治疗后7、14、21 d的DASI评分分别为(30.44±5.68)、(5.28±2.31)、(2.04±1.12)分,均明显低于治疗前[(52.34±7.45)分],差异均有统计学意义(P<0.05)。相较治疗前、治疗7 d、治疗14 d,治疗21 d后的DASI50(100.00%)、DASI75(100.00%)、DASI90(90.00%)的改善比率最高,差异均有统计学意义(P<0.05)。TEN患者治疗后7、14、21 d的血清TNF-α水平分别为(22.73±5.58)、(15.99±4.60)、(4.44±1.10)pg/mL,均低于治疗前[(33.63±17.36)pg/mL],差异均有统计学意义(P<0.05)。TEN患者体温下降时间为(2.49±0.81)d,皮疹控制时间为(5.19±1.90)d,住院时间为(11.92±4.20)d。治疗期间患者未出现终止治疗或失访,均未出现急性不良反应,随访期间病情未见复发,定期复查结果显示并无合并症、活动性肝炎与结核疾病等。结论rhTNFR:Fc作为治疗TEN疾病的药物其疗效随治疗时间延长而提高,可降低血清TNF-α水平与DASI评分,且安全性较高。

重组人血管内皮抑制素联合脑部放疗与单纯脑部放疗治疗实体肿瘤脑转移的疗效及预后

目的探究重组人血管内皮抑制素(rh-Endostatin)联合脑部放疗(WBRT)与单纯WBRT治疗实体肿瘤脑转移的疗效及预后。方法回顾性分析2020-03—2022-11于资阳市人民医院肿瘤科治疗的94例患者的临床资料,根据治疗方式分为试验组和对照组,采用WBRT治疗的46例患者纳入对照组,采用rh-Endostatin联合WBRT治疗的48例患者纳入试验组,治疗结束1个月后比较临床疗效及肿瘤标志物指标[癌胚抗原(CEA)、癌抗原(CA125)、糖类抗原(CA19-9)、血管内皮生长因子(VEGFA、VEGFB、VEGFC)]、免疫功能(CD3^(+)、CD4^(+)、CD8^(+)、CD4^(+)/CD8^(+))、脑水肿体积和脑血流动力学参数(大脑前动脉平均血流速度和搏动指数)、生存质量[欧洲癌症研究治疗组织生命质量测定量表(EORTCQLQ)]、神经认知水平[简易精神状态量表(MMSE)],比较2组患者治疗期间放射性损伤,通过制作生存曲线分析患者预后情况。结果试验组患者治疗结束1个月时临床总有效率高于对照组(47.92%比26.09%),差异有统计学意义(P<0.05);2组患者治疗结束1个月时肿瘤标志物CEA、CA125、CA19-9、VEGFA、VEGFB、VEGFC水平,CD3^(+)、CD4^(+)、CD4^(+)/CD8^(+)水平,脑水肿体积、搏动指数均较治疗前显著降低(P<0.05),试验组患者肿瘤标志物水平和脑水肿体积显著低于对照组患者(18.76±2.54比20.78±3.12,26.78±4.98比33.12±5.19,31.34±3.12比37.24±3.98,90.76±16.97比121.78±18.12,83.78±11.02比105.23±12.19,65.34±9.12比83.23±10.98,144.58±71.55比203.87±70.59),CD3^(+)、CD4^(+)、CD4^(+)/CD8^(+)水平显著升高(59.67±4.33比55.41±4.02,31.22±3.34比27.74±3.79,1.17±0.22比0.96±0.15),差异均有统计学意义(P<0.05);2组患者CD8^(+)水平、大脑前动脉平均血流速度、EORTC各维度评分和MMSE评分相较治疗前均明显升高(P<0.05),试验组患者CD8^(+)水平明显低于对照组(27.02±3.08比28.92±3.31),大脑前动脉平均血流速度(56.67±3.33比55.01±2.06)、EORTC各维度评分和MMSE评分(26.58±1.11比25.87±1.17)明显高于对照组,差异均有统计学意义(P<0.05);2组患者治疗期间均发生轻微放射性损伤,差异无统计学意义(P>0.05);治疗后试验组患者中位总生存期(OS)和颅内疾病无进展期(iPFS)均明显长于对照组患者(9.5比7.5,8.0比6.0),差异有统计学意义(P<0.05)。结论rh-Endostatin联合WBRT治疗实体肿瘤脑转移患者可有效改善近期疗效,抑制肿瘤进展,改善脑血流,维持认知功能及生存质量,保护免疫功能。该方案安全性良好,可一定程度上延长患者的OS和iPFS。

Inhibition of telomerase with human telomerase reverse transcriptase antisense increases the sensitivity of tumor necrosis factor-α-induced apoptosis in prostate cancer cells

Aim： To investigate the effect of inhibition of telomerase with human telomerase reverse transcriptase （hTERT） antisense on tumor necrosis factor-α （TNF-α-induced apoptosis in prostate cancer cells （PC3）. Methods： Antisense phosphorothioate oligodeoxynucleotide （AS PS-ODN） was synthesized and purified. Telomerase activity was measured using the telomeric repeat amplification protocol （TRAP） and polymerase chain reaction enzyme-linked immunoassay （PCR-ELISA）. hTERT mRNA was measured by reverse transcription PCR （RT-PCR） assay and gel-image system, hTERT protein was detected by immunochemistry and flow cytometry. Cell viability was detected by 3-（4, 5-dimethylthiazol-2-yl）-2,5-Diphenyltetrazolium （MTT） assay. Cell apoptosis was observed by morphological method and determined by flow cytometry. Results： The telomerase activity decreased with time after hTERT AS PS-ODN treatment. The levels of hTERT mRNA decreased with time after hTERT AS PS-ODN treatment, which appeared before the decline of the telomerase activity. The percentage of positive cells of hTERT protein declined with time after hTERT AS PS-ODN treatment, which appeared after the decline of hTERT mRNA. There was no difference in telomerase activity, hTERT mRNA and protein levels between hTERT sense phosphorothioate oligodeoxynucleotide （S PS-ODN） and the control group. The cell viability decreased with time after hTERT AS PS-ODN combined with TNF-α treatment. The percentage of apoptosis increased with time after hTERT AS PS-ODN combined with TNF-α treatment. There was no difference in cell viability and the percentage of apoptosis between hTERT S PS-ODN and the control group. Conclusion： hTERT AS PS-ODN can significantly inhibit telomerase activity by downregulating the hTERT mRNA and protein expression, and inhibition of telomerase with hTERT antisense can enhance TNF-α- induced apoptosis of PC3 cells.

重组人血管内皮抑制素注射液联合GP化疗方案治疗晚期NSCLC的可行性

目的观察重组人血管内皮抑制素注射液联合GP化疗方案治疗晚期非小细胞肺癌(NSCLC)的可行性。方法回顾性选取2020年5月—2022年12月皖北煤电集团总医院肿瘤内科收治的晚期NSCLC患者80例,将采用GP化疗方案治疗者纳入GP化疗组(n=40),重组人血管内皮抑制素注射液联合GP化疗方案治疗者纳入联合用药组(n=40),2组均以3周为1个治疗周期,共治疗2个周期。比较2组患者近期疗效,用药前后实验室指标、生活质量评分,不良反应及1年存活率。结果联合用药组客观缓解率为85.00%,高于GP化疗组的52.50%(χ^(2)=9.833,P=0.002)。用药2个周期后,2组血清癌胚抗原、糖类抗原125水平低于用药前,凝血酶时间、活化部分凝血活酶时间短于用药前,凝血酶原时间长于用药前,血浆纤维蛋白原、D-二聚体水平高于用药前,且联合用药组各指标优于对照组(P<0.05或P<0.01);2组生理功能、躯体功能、精神状态、社会关系评分高于用药前,且联合用药组高于GP化疗组(P<0.01)。联合用药组与GP化疗组不良反应总发生率分别为22.50%、15.00%,组间比较差异无统计学意义(χ^(2)=0.738,P=0.390)。联合用药组1年存活率为65.00%,高于GP化疗组的42.50%(χ^(2)=4.073,P=0.044)。结论晚期NSCLC治疗中采用重组人血管内皮抑制素注射液联合化疗药物的可行性较高,可有效降低肿瘤标志物水平,延缓病情进展,提高患者生活质量及1年存活率,且具有一定安全性。

STAT3 as a target for inducing apoptosis in solid and hematological tumors

Studies in the past few years have provided compelling evidence for the critical role of aberrant Signal Transducer and Activator of Transcription 3 （STAT3） in malignant transformation and tumorigenesis. Thus, it is now generally accepted that STAT3 is one of the critical players in human cancer formation and represents a valid target for novel anticancer drug design. This review focuses on aberrant STAT3 and its role in promoting tumor cell survival and sup- porting the malignant phenotype. A brief evaluation of the current strategies targeting STAT3 for the development of novel anticancer agents against human tumors harboring constitutively active STAT3 will also be presented.