艾迪注射液配合化疗治疗晚期非小细胞肺癌的临床观察

为研究艾迪注射液对晚期非小细胞肺癌的治疗作用及对化疗所致毒副作用的影响 ,将 76例晚期非小细胞肺癌患者随机分为两组 ,分别按MVP方案化疗 ,MVP化疗基础上加用艾迪注射液 ,按WHO的统一评价标准评价两组的近期疗效和毒副作用并进行比较。结果发现两组间的近期疗效差异无显著性 (P >0 0 5 ) ,但是联合使用艾迪注射液组的Ⅱ度和Ⅱ度以上的胃肠道反应、白细胞下降率、血小板下降率均显著低于单纯MVP化疗组 (均P <0 0 5 )。初步研究结果提示 ,艾迪注射液能够显著减轻化疗药物引起的胃肠道反应 ,对骨髓具有保护作用。

mTOR信号传导通路及其相关药物与非小细胞肺癌

哺乳动物雷帕霉素靶蛋白（mTOR）处于细胞生长繁殖、细胞周期调控、生物合成、细胞迁移等信号通路调控的中心位置，因此在肿瘤以及肺癌的发生、发展，特别是在治疗和预后中具有重要的作用。mTOR磷酸化激活后，通过调控4EBP和P70S6K两条不同的下游通路，分别控制特定亚组mRNA的翻译，进而影响生物合成。对非小细胞肺癌来讲，mTOR能够加快细胞周期G1-S期的转换，促进细胞增殖。雷帕霉素、西罗莫司、依维莫司等mTOR信号通路的阻断药物在肺癌的治疗中显示出了希望。

Programmed cell death ligand-1(PD-L1) expression by immunohistochemistry: could it be predictive and/or prognostic in non-small cell lung cancer?

Blockade of immune checkpoints has recently emerged as a novel therapeutic strategy in various tumors. In particular,monoclonal antibodies targeting programmed cell death 1(PD-1) or its ligand(PD-L1) have been most studied in lung cancer,and PD-1 inhibitors are now established agents in the management of non-small cell lung cancer(NSCLC). The reports on highprofile clinical trials have shown the association of PD-L1 expression by immunohistochemistry(IHC) with higher overall response rates to the PD-1/PD-L1 axis blockade suggesting that PD-L1 expression may serve as a predictive marker. Unfortunately,however, each PD-1 or PD-L1 inhibitor is coupled with a specific PD-L1 antibody, IHC protocol and scoring system for the biomarker assessment, making the head-to-head comparison of the studies difficult. Similarly, multiple clinical series that correlated PD-L1 expression with clinicopathologic and/or molecular variables and/or survival have reported conflicting results.The discrepancy could be explained by the differences in ethnicity and/or histologic types included in the studies, but it appears to be attributed in part to the differences in PD-L1 IHC methods. Thus, orchestrated efforts to standardize the PD-L1 IHC are warranted to establish the IHC as a predictive and/or prognostic biomarker in NSCLC.

Effects of Tai Chi Chuan training on cellular immunity in post-surgical non-small cell lung cancer survivors:A randomized pilot trial

Background： Although emerging evidence points to benefits from Tai Chi Chuan （TCC） in improving immune system function, its effects on cellular immune responses remain under-studied. The objective of this study was to evaluate the effects of TCC training on cellular immunity in non-small cell lung cancer patients. Methods： A 2-group randomized trial design in which post-surgical, non-small cell lung cancer survivors were randomly assigned to a TCC training group （n = 16） or a control group （n = 16）. The participants in the TCC group completed a 16-week intervention. The main immune response outcome measures assayed included the ratio of T-helper cells/T-suppressor cells （CD4＋：CD8＋ ratio） and complement regulatory proteins status （CRPs; CD55 and CD59）. Using repeated measures ANOVA, the data were analyzed for the participants who completed the study （n = 27）. Results： At 16 weeks, the TCC participants showed a significantly lower increment in the expression of CD55 （p 〈 0.05） as compared to the control group. No significant between-group differences were found in the CD4＋：CD8＋ ratio or CD59 expression. There were also no significant correlations among the changes in CRPs or T lymphocyte subpopulations, either. Conclusion： A 16-week TCC intervention caused no alterations in CD4＋：CD8＋ ratio, but significantly attenuated CD55 expression among post- surgical non-small cell lung cancer survivors.

Phase Ⅲ Clinical Trials of the Cell Differentiation Agent-2 （CDA-2）： Therapeutic Efficacy on Breast Cancer, Non-Small Cell Lung Cancer and Primary Hepatoma

OBJECTIVE The objective of this study was to explore the effect of CDA-2, a selective inhibitor of abnormal methylation enzymes in cancer cells, on the therapeutic efficacy of cytotoxic chemotherapy. METHODS Advanced cancer patients, all of whom had previously undergone chemotherapy, were randomly divided into 2 groups, one receiving chemotherapy only as the control group, and the other receiving CDA-2 in addition to chemotherapy as the combination group. The therapeutic efficacies and the toxic maniestations of the 2 groups were compared based on the WHO criteria. RESULTS Of 454 cancer patients enrolled in phase Ⅲ clinical trials of CDA-2, 80, 188, and 186 were breast cancer, NSCLC, and primary hepatoma patients, respectively. Among them 378 patients completed treatments according to the protocols. The results showed that the overall effective rate of the combination group was 2.6 fold that of the control group, 4.8 fold in the case of breast cancer, 2.3 fold in the case of primary hepatoma, and 2.2 fold in the case of NSCLC. Surprisingly, the combination therapy appeared to work better for stage Ⅳ than stage Ⅲ patients. CDA-2 did not contribute additional toxicity. On the contrary, it reduced toxic manifestations of chemotherapy, particularly regarding white blood cells, nausea and vomiting. CONCLUSION Modulation of abnormal methylation enzymes by CDA-2 is definitely helpful to supplement chemotherapy. It significantly increased the therapeutic efficacy and reduced the toxic manifestation of cytotoxic chemotherapy on breast cancer and NSCLC.

Relationship between the Expression of NF-kappa B and Apoptosis in Non-Small Cell Lung Cancer

OBJECTIVE To study the expression of the nuclear factor kappa B (NF-kappa B) in non-small cell lung cancer (NSCLC),to explore the apoptotic ratio in NSCLC related to different NF-kappa Bs,and to understand the clinical significance of NF-kappa B in NSCLC apoptosis. METHODS NF-kappa B expression in 45 new samples of NSCLC,collected during a period from October to December,2005,was assayed using Western blots,and the apoptotic ratio of NSCLC was determined by the Tunel method. RESULTS Of the 45 patients,the average relative expression of NF-kappa B was 0.6047±0.3572.The expression of NF-kappa B was higher in the poorly differentiated lung cancer cells than in the well-differentiated tumors(P<0.05).The apoptotic ratio was 56.4% in the lung cancer cells with higher NF-kappa B expression,and was 76.7% in those with lower NF-kappa B expression(P<0.05). CONCLUSION The expression of NF-kappa B is correlated with the differentiation of NSCLC.NF-kappa B inhibits apoptosis in NSCLC.As a transcription factor,NF-kappa B plays a very important role both in formation and in development of NSCLC. NF-kappa B might serve as a target for NSCLC gene therapy.

Influence of Vein Injury in Different Methods of Chemotherapy in Mice

OBJECTIVE To explore the effects of cisplatin chemotherapy via the vein and the abdominal cavity on the functions of endothelial cells of mice with non-small cell lung cancer. METHODS 75 mice were divided equally into a untreated group, a group treated with chemotherapy （cisplatin： 2 mg/kg, 0.5 mL） via an intravenous method and a group treated with chemotherapy （cisplatin： 2 mg/kg, 0.5 mL） via an intraperitoneal method; changes in the morpholoqy and ultrastructure of vein endothelial cells were observed.

Multi-Targeted Therapies in Non-Small Cell Lung Cancer

Current treatment modalities provide limited improvement in the natural course of lung cancer, and prognosis remains poor. Lung cancer is a malignancy with great molecular heterogeneity. The complexity of the signaling process leading to cancer cell proliferation and to the neoplastic phenotype supports the necessity of interfering at different stages to avoid cancer cell resistance to therapy. For this reason, new strategies for the simultaneous inhibition of multiple molecular targets are being pursued.

INVESTIGATION OF REGULATORY T CELLS IN PATIENTS WITH NON-SMALL CELL LUNG CANCER

Objective To evaluate the prevalence of CD4 ^＋ CD25^high regulatory T cells （ Treg cells） in the peripheral blood mononuclear cells （PBMC） and tumor-infiltrating lymphocytes （TIL） of patients with non-small cell lung cancer （NSCLC） and to investigate immunosuppression to the progression of cancer. Methods Peripheral blood and tumor tissues were collected from 20 patients with NSCLC at the time of surgery. None of the patients received surgery, radiotherapy, chemotherapy, or other medical interventions before this study. Cancer stages of the patients were Ⅰ-Ⅲ A. Venous blood samples were obtained from 20 health donors. PBMC were isolated from blood samples by differential centrifugation over Ficoll-Hypaque. TILs were isolated from tumors by differential centrifugation over Ficoll-Hypaque and Percoll. Percentage of CD4^＋ CD25^highTr/CD4＋T in PBMC and TIL was assessed by the flow cytometry. Results The percentage of CD4^ ＋ CD25high Tr/ CD4 ^＋T in PBMC [ （4. 87 ± 1.22 ） % ] of NSCLC patients was significantly higher than that in healthy donors [ （ 2.36 ± 0. 72 ） % ] （ P 〈 0.01 ）. The percentage of CD4^＋ CD25^highTr/ CD4^＋ T in PBMC [ （5.40 ± 1.20） % ] of NSCLC patients in stage Ⅱ-Ⅲ A was significantly higher than that in stage Ⅰ [ （3. 87 ± 0. 22 ） % ] （ P 〈 0. 01 ）. The percentage of CD4 ＋ CD25hiShTr/ CD4 ＋ T in TIL[ （ 8. 66 ±0. 76） % ] of NSCLC patients in stage Ⅱ-Ⅲ A was significantly higher than that in stage Ⅰ [ （ 7. 04 ± 0. 80） % ] （ P 〈 0. 01 ）. Conclusion The prevalence of CD4 ^＋ CD25^highTreg cells in PBMC and TIL of NSCLC patients was significantly higher than that in healthy donors. These Treg cells may be preventing appropriate antitumor immune responses. The population of CD4^ ＋ CD25^highTreg cells in PBMC and TILs of NSCLC patients with Ⅱ-Ⅲ A stage was significantly higher than that of NSCLC patients with Ⅰ stage. These Treg cells may facilitate development of tumors.

The influence of autologous cytokine-induced killer cell treatment on the objective efficacy and safety of gefitinib in advanced non-small cell lung cancer

Objective The aim of the study was to observe the influence of autologous cytokine-induced killer cell （CIK） treatment on the objective efficacy and safety of gefitinib in advanced non-small ceil lung cancer （NSCLC）. Methods Sixty-six patients with NSCLC received gefitinib as second-line treatment. They were randomly divided into 2 groups, and informed consent forms were signed before grouping. Gefitinib was administrated to the control group, and autologous CIK treatment was added to the observation group. The objective treatment and adverse reactions were evaluated in both groups. Results The objective response rate （ORR） and the disease control rate （DCR） of the observation group were slightly higher than those of the control group, although no statistical differences were found between the 2 groups （P 〉 0.05）. The incidences of diarrhea, fatigue, anorexia, oral ulcers, and myelosuppression in the observation group were much lower than those in the control group （P 〈 0.05）. However, there were no statistical differences between the incidences of skin rash, and liver and kidney toxicities （P 〉 0.05）. Conclusion Autologous CIK in combination with gefitinib is effective as second-line treatment fore ad- vanced NSCLC, and can significantly reduce adverse reactions and improve the objective efficacy.

Biallelic Inactivation of hMLH1 by Hypermethylation and Loss of Heterozygosity in Non-Small Cell Lung Cancer

OBJECTIVE To investigate the mechanism of hMLH1 deregulation in non-small cell lung cancer （NSCLC）. METHODS A genetic and epigenetic study of the hMLH1 gene was performed using surgical primary tumors from 40 NSCLC patients and their corresponding noncancerous tissues. The molecular alterations examined included promoter methylation by Hpa Ⅱ/Msp Ⅰ- based PCR analysis, loss of heterozygosity （LOH） by D3S1621 locus PCR-electrophoresis-silver staining, as well as the loss of protein expression by immunohistochemical analysis. RESULTS The frequencies of hypermethylation, LOH and loss of protein expression of hMLH1 were 67.5% （27/40）, 65% （26/40） and 72.5% （29/ 40）, respectively. Among 26 hMLH1 gene LOH （＋） cases, 21 （80.8%） showed hypermethylation, which was significantly higher than the group of LOH （-）. The frequency of the double inactivation of the hMLH1 gene by hypermethylation and LOH related to a loss of protein expression of 72.4% （21/29）. CONCLUSION Biallelic inactivation of the hMLH1 gene by hypermethylation and LOH most likely will cause loss of hMLH1 protein expression and play an important role in the development of NSCLC. Therefore, controlling and monitoring for hypermethylation of the hMLH1 gene may be partially useful for treatment and early diagnosis of NSCLC.

Argyrophilic nucleolar organizer region in MIB-1 positive cells in non-small cell lung cancer: clinicopathological significance and survival

Objective： To evaluate the relation between argyrophilic nucleolar organizer region （AgNOR）-associated proteins and clinicopathological parameters and survival in non-small-cell lung cancer （NSCLC）. Methods： A total of 207 surgical specimens diagnosed as NSCLC were included in this study. Double-staining procedures were performed using antigen Ki-67 （clone MIB-1） and silver nitrate by immunohistochemical and AgNOR-staining methods. Results： The AgNOR area in MIB-l-positive cells of NSCLC is related to clinicopathological parameters under the TNM （tumor, node, and metastasis） system. The survival of patients with small AgNOR area in MIB-1-positive cells is better than that of patients with large AgNOR area. Molecular, biological （AgNOR area in MIB-l-positive cells）, and clinicopathological （greatest tumor dimension, metastases to regional lymph nodes, histology, and differentiation） parameters are independent prognostic factors of NSCLC.Conclusion： The AgNOR area in MIB- 1-positive cells is related to clinicopathological parameters and survival in NSCLC.

The preparation of anti-hnRNP A2/B1 polyclonal antibody and its potential application in non-small cell lung cancer

Objective： In order to evaluate potential application for diagnosis and prognosis of non-small cell-lung cancer （NSCLC）, as well as to determine its role in the pathogenesis of the disease, we prepared anti-human hnRNPA2/B1 potyclonal antibody. Methods： Prokaryotic expression vector of pET28a （＋）-hnRNP A2/B1 was constructed and bansformed into E.coli BL21. The recombinant protein induced by IPTG was purified and injected to rabbits for antibody preparation. Expression of hnRN P A2/B1 was examined in 45 tissues of NSCLC and 16 inflammatory pseudotumor tissues of lung by immunohistochemistry with the antibody. The commercial hnRNP A2/B1 monoclonal antibody was used as a controI.Results： （1） Polyclonal an-tibody against hnRNP A2/B1 with high title was obtained. （2） The positive staining in NSCLC tissues was 62.22%, which was substantially higher than that in normal tissues （40%, P = 0.035） or inflammatory pseudotumor tissues （31.25%, P=0.033）. （3） Expression of hnRNP A2/B1 positively correlated with age and the history of smoking, whereas it negatively correlated with differentiation staging of tumors. （4） Follow-up study showed that the survival time of patients with positive staining was significantly shorter than that of patients without hnRNP A2/B1 expression （P=0.048）. Conclusion： It is successful to make the recombinant protein and prepare the polyclonal antibody agonist human hnRNP A2/B1. It may be a valuable marker for the diagnosis and prognosis of NSCLC. Our results provide a basis for further study in clinical application.

Inhibitory effect of toremifene monotherapy or combined with gemcitabine on A549 human lung adenocarcinoma cells

Objective： The aim of this study was to investigate the effect of toremifene on A549 human lung adenocarci- noma cells, and its sensibilization with gemcitabine, so that to provide a new clinical approach for non-small-cell lung cancer （NSCLC）. Methods： A549 cells were seeded into 96-well plates and exposed to different agents （gemcitabine or gemcitabine with toremifene）. The cytotoxicity of each agent was evaluated by MTT, cell cycle and apoptotic rate were detected by flow cytometry （FCM）. Results： 1. By using FCM, we found A549 cells in S and G2/M phases with toremifene decreased but increased in G0/G1 phase. The higher concentration of toremifene, the more decreased was when compared with the control group. 2. FCM showed toremifene＇s apoptosis effect on A549 cells increased with its increasing dose. 3. By MTT, toremifene had no cytotoxic effect on A549 cells at the concentration of 5 or 2.5 pmol/L. The IC5o of gemcitabine to A549 was 34.51 tJmol/L, and the combined group was 13.59 pmol/L. Conclusion： Toremifene could inhibit the growth of A549 human lung adenocarcinoma cells. Toremifene combined with gemcitabine showed significantly remarkable chemotherapy sensibilization on A549 human lung adenocarcinoma cells.

Chimeric antigen receptor-modified T cells for the immunotherapy of patients with EGFR-expressing advanced relapsed/refractory non-small cell lung cancer

The successes achieved by chimeric antigen receptor-modified T （CAR-T） cells in hematological malignancies raised the pos- sibility of their use in non-small lung cancer （NSCLC）. In this phase I clinical study （NCT01869166）, patients with epidermal growth factor receptor （EGFR）-positive （〉50% expression）, relapsed/refractory NSCLC received escalating doses of EGFR-targeted CAR-T cell infusions. The EGFR-targeted CAR-T cells were generated from peripheral blood after a 10 to 13-day in vitro expansion. Serum cytokines in peripheral blood and copy numbers of CAR-EGFR transgene in peripheral blood and in tissue biopsy were monitored periodically. Clinical responses were evaluated with RECISTI.1 and im- mune-related response criteria, and adverse events were graded with CTCAE 4.0. The EGFR-targeted CAR-T cell infusions were well-tolerated without severe toxicity. Of 11 evaluable patients, two patients obtained partial response and five had stable disease for two to eight months. The median dose of transfused CAR＋ T cells was 0.97x 10^7 cells kg J （interquar- tile range （IQR）, 0.45 to 1.09x 10^7 cells kg 1）. Pathological eradication of EGFR positive tumor cells after EGFR-targeted CAR-T cell treatment can be observed in tumor biopsies, along with the CAR-EGFR gene detected in tumor-infiltrating T cells in all four biopsied patients. The EGFR-targeted CAR-T cell therapy is safe and feasible for EGFR-positive advanced re- lapsed/refractory NSCLC.

Effect of ginseng polysaccharides and dendritic cells on the balance of Th1/Th2 T helper cells in patients with non-small cell lung cancer

OBJECTIVE: To investigate the effect of thorascopic administration of ginseng polysaccharides(GPS)plus dendritic cells(DC) on T helper cell type 1/T helper cell type 2(Th1/Th2) balance in patients with non-small cell lung cancer(NSCLC).METHODS: A total of 96 NSCLC patients were divided evenly into two groups. The control group was treated with DCs alone and the treatment group was treated with DCs plus GPS. After DCs and GPS were administered thoracoscopically, once a week,4 times for 30 days, the patients' quality of life was measured with the Functional Assessment of Can-cer Treatment-Lung(FACT-L) questionnaire before and after treatment. Serum interferon-γ(INF-γ), interleukin-4(IL-4), IL-2 and IL-5 were examined before and after treatments.RESULTS: The level of Th1 cytokines(INF-γ, IL-2)and the ratio of Th1/Th2 cytokines(INF-γ/IL-4, IL-2/IL-5) increased in both treatment groups, while Th2cytokines(IL-4, IL-5) and FACT-L scores decreased(P<0.01). Furthermore, after treatment Th1 cytokines(INF-γ, IL-2) and the ratio of Th1/Th2 cytokines(INF-γ/IL-4, IL-2/IL-5) were higher in the DCs +GPS group than in the control group(P<0.05). Conversely, FACT-L scores and Th2 cytokines(IL-4, IL-5)were higher in the control group than in the DCs +GPS group(P<0.05).CONCLUSION: The treatment regime of DCs plus GPS had a greater effect on NSCLC patients' immune function as compared with DCs alone. This was evident by increased expression of Th1 cytokines(INF-γ, IL-2) and the ratio of Th1/Th2(INF-γ/IL-4, IL-2/IL-5), as well as by decreased FACT-L scores and the expression of Th2 cytokines(IL-4,IL-5).

Influence of Erbanxiao solution on inhibiting angiogenesis in stasis toxin stagnation of non-small cell lung cancer

OBJECTIVE： To investigate the effects and mecha- nisms of Erbanxiao solution in inhibiting tumor an- giogenesis. METHODS： We observed the effects and mecha- nisms of Chinese medicines on inhibiting tumor an- giogenesis and studied the theories and results of treatment. Sixty patients with lung cancer were ran- domized into two groups （n=30）. Patients in the control group were given compound Kushen injec- tion, and patients in the treatment group were giv- en Erbanxiao solution. The effect of Erbanxiao solu- tion on vascular endothelium growth factor （VEGF）, basic fibroblast growth factor （bFGF）, and tumor ne- crosis factor-α （TNF-α） was observed. RESULTS： The effective rate of the treatment group was 60% while the control group was 36%. There was a significant difference between the twogroups （P〈0.05）. VEGF, bFGF, and TNF-a levels of the two groups were significantly different before and after treatment （P〈0.01）. These Traditional Chi- nese Medicines significantly inhibited tumor angio- genesis, possibly by changing levels of VEGF, bFGF, and TNF-α. CONCLUSION： It is necessary to further explore the potential of Traditional Chinese Medicine in the treatment of angiogenesis in tumor patients.