Objective To investigate the expression of CD4+CDhi25CDlow127 regulatory T cells (Treg cell), IL-2, IL-10 and IFN-γin peripheral blood of patients with different TNM stages of non-small cell lung cancer( NSCLC) , and to evaluate their correlation. Methods The proportion of CD4+CDhi25 CDlow127 Treg cells to CD4+cell in peripheral blood of 55 NSCLC patients were determined by flow cytometry, and levels of IL-2, IL-10 and IFN-γ were measured by ELISA. The proportion of CD4+CDhi25 CDlow127 Treg cells, IL-2, IL-10 and IFN-γ of patients with different TNM stages of NSCLC were compared, and correlation analysis was carried on. Results The proportion of CD4+CDhi25 CDlow127 Treg cells increased obviously together with the increasing of TNM stages of NSCLC(P<0. 05),so did the level of IL-10 (P<0. 05). On the other hand, the level of IL-2 and IFN-γdecreased apparently with the increasing of TNM sta-ges of NSCLC(P<0. 05). CD4+CDhi25CDlow127Treg cells and IL-10 were positively correlated(P <0. 05). IL-2 and IFN-γ were positively correlated too(P<0. 05). CD4+CDhi25CDlow127Treg cells and IL-10 were all negatively correlated with IL-2 and IFN-γ(P<0. 05). Conclusion The increasing of CD4+ CDhi25CDlow127Treg cells and IL-10 and the de-creasing of IL-2 and IFN-γ are possibly one of the reasons of antitumor immunity suppression in terminal NSCLC patients. The surveillance for them can be reference indicators to evaluate antitumor immunity status of terminal lung cancer patients.

OBJECTIVE:To explore the effect of alendronte sodium and vitamin D3 on the T cell immune indexes in patients with different genders and ages with advanced lung cancer. METHODS:76 patients with advanced lung cancer were divided into non-small cell lung cancer(NSCLC)group(n=40)and small cell lung cancer(SCLC)group(n=36),and another 38 healthy vol-unteers were control group. Based on palliative care,all patients received Alendronte sodium and vitamin D3 tablet 1 tablet,po,qd, for 1 month. T cell immune indexes(CD4+,CD8+and Treg levels)in 2 groups before and after treatment,and when volunteers enroll-ing the control group were compared;and the incidence of adverse reactions were observed. RESULTS:Compared with control group,there were significant differences in CD4+,CD8+,CD4+/CD8+and Treg levels(P<0.05);but there was no significant differ-ence between NSCLC group and SCLC group(P>0.05). After treatment,CD4+ and CD4+/CD8+ levels in NSCLC group and SCLC group increased,changes of male were greater than female,CD8+and Treg levels decreased,changes of male were greater than fe-male,except for CD8+ level in NSCLC group (P=0.33),there were significant differences in others (P<0.05);CD4+ and CD4+/CD8+ levels in patients ≤50 years old increased more than patients >50 years old,CD8+ and Treg levels decreased more than pa-tients >50 years old,the differences were statistically significant (P<0.05). No obvious adverse reactions were found. CONCLU-SIONS:Alendronte sodium and vitamin D3 can effectively improve the T cell immune indexes in patients with advanced lung cancer, especially for male and patients≤50 years old,which can be considered as conventional adjuvant drugs for patients with lung cancer.

Objective To investigate the effects of co-transfection of the plasmid containing the wildtype (wt) INK4a/ARF gene on the chemosensitivity of human lung adenocarcinoma cell line A549. Methods Recombinant eukaryotic expression vectors pcDNA3-p16 INK4a and pcDNA3-p14 ARF were co-transfected into A549 cells by cationic liposome, in which the site of both genes was lost. By RT-PCR, immunocytochemistry and Western blotting analysis, G418 positive clone was obtained. The parental cell and negative control cell with plasmid pcDNA3-LacZ were used as controls. The 50% inhibition concentration (IC 50 ) of five commonly-used chemotherapy drugs and the apoptosis index (AI) of IC 50 were analysed respectively. Results Compared with the parental cells and negative control cells, the influence of the five drugs on IC50 was different. The values of IC 50 of doxorubicin and cisplatin decreased significantly but that of taxol, topotecan and vinorelbine remained unchanged. The apoptosis index of doxorubicin and cisplatin was much higher than that of other three drugs. Conclusion The chemosensitivity of a part of anti-cancer drugs could be changed by transfection of exogenous INK4a/ARF gene into lung adenocarcinoma cell A549.

Objective To evaluate the therapeutic efficacy and side effects of cyclopnosphamide, adriamycin, cisplatin (CAP) or CAP combined with verapamil(VPL) in chemotherapy of patients with advanced lung adenocarcinoma. Methods A total of 56 patients(male: 27, female: 29, average age: 48 years old, age range: 32-78 years old) with stage Ⅲb/Ⅳ lung adenocarcinoma confirmed pathologically from April 1998 to December 2001 were divided into two groups: group A(30 patients), treated with cyclophosphamide (CTX, 600 mg?m -2 ?d -1 ) on days 1 and 8, adriamycin (ADM, 40 mg/m 2) on day 1, and cisplatin (CDDP, 30 mg?m -2 ?d -1 ) on days 1, 2 and 3 and group B(26 patients), treated with the same dose of CTX, ADM and CDDP and an additional oral VPL treatment (60 mg t.i.d . on days 1-7). Each of the 3 cycles was repeated every 21 days. Results The patients in group A had a lower response rate (26.7% vs 38.5%, P

Objective To observe the efficacy of CT guided implantation of iodine 125 particles combined with high-frequency hyperthermia in the treatment of the advanced lung cancer .Methods From 2011 November to 2013 March ,60 patients with ad-vanced lung cancer were randomly divided into treatment group :high frequency hyperthermia combined with iodine 125 particle im-plantation(30 cases) ,the control group :the iodine 125 particle implantation group(30 cases) .Thermotherapy was performed 3 -5 days after the implantation using NRL-004 type hyperthermia instrument made by Jilin Maida company in China ,every other day once time ,a total of six times ,60 minutes each time .The iodine 125 particle implantation was performed according to the treatment planning system(TPS) formulation by CT guided and the dosage was 90 Gy .The evaluation of treatment effect was conformed by CT scans after two months of treatment .The side effects was assessed too .Results The total effective rate was 96 .7% in the treatment group ,the control group was 80 .0% ,there was a statistically significant difference (P< 0 .05) .The local pain in chest wall remission rate was 75 .0% (6/8) and 50 .0% (5/10) in the treatment group and the control group(P>0 .05) .The side effects of the treatment was slight including pneumothorax and particle migration and so on .Conclusion Iodine 125 particles implantation is an effective treatment in the treatment of advanced lung cancer and can obtained better effects combined with high-frequency hy-perthermia .The side effects of the treatment is slight for two groups and no death related treatment .

Objective To study the effect of the recombinant matrilysin (rMMP-7) and its antisense oligonucleotide transfected by liposome (LIPO-MAON) on the proliferation of lung adenocarcinoma cell line A549 and human umbilical vein endothelial cells (HUVEC). Methods Antisense oligonucleotide of matrilysin was constructed by liposome transfection. The proliferation of HUVEC and A549 was detected by MTT assay in case of rMMP-7 or LIPO-MAON existence. The expression of MMP-7 mRNA in both HUVEC and A549 was examined by semi-quantitative RT-PCR assay. Results The proliferation of both HUVEC and A549 cell line was accelerated by high level of rMMP-7 (0.75, 1.0 ?g/ml), and the inhibited proliferation was only found in A549 cell line treated with high concentration of LIPO-MAON (7.5, 10 nmol/ml), but not in HUVEC. By RT-PCR assay, no expression of MMP-7 mRNA was found in HUVEC; however, enhanced or decreased expression of mRNA were found when A549 cell line was treated respectively with rMMP-7 or LIPO-MAON (P

Indoleamine 2,3-dioxygenase(IDO)is the crucial rate-limiting enzyme of tryptophan outside the liver in the body and is considered as an immune defense mechanism in tumor microenvironment.IDO is expressed in a number of cells including cancer cell,dendretic cell and so on.In tumor microenvironment,the overexpression of IDO can lead to the exhaustion of tryptophan and induce the proliferation of regulatory cells,which can prevent the proliferation,diffenentiation and activity of T cells and result in the immune tolerance to tumors.

BACKGROUNDp16INK4a and p14ARF, encoded by gene INK4a/ARF located at chromosome 9p21, are cyclin dependent kinase (CDK) inhibitors. Both p16INK4a and p14ARF are cell cycle regulatory proteins and play an important role in Rb and p53 passways respectively. In this study, wild-type INK4a/ARF gene was transfected into human lung adenocarcinoma cell line A549, in which this gene site was lost, and the effects on the cell's biological behavior were investigated.

METHODSThe recombinant eukaryotic expression plasmids pcDNA3-p16INK4a and pcDNA3-p14ARF were transfected into A549 by cationic liposome method. By RT-PCR, immunocytochemistry and Western blot after G418 selection, A549 cells that could stably express p16INK4a and p14ARF were obtained. As a control, the parental cell and negative control cell with plasmid pcDNA3-LacZ were used. Inhibition of proliferation was measured by MTT assay. The cell growth curve was drawn according to cell counts. Cell cycle distribution was measured by flow cytometry (FCM), the apoptosis indexes were observed at the same time. The colony formation rate was counted by staining the cells with Coomassie brilliant blue.

RESULTSThe introduction of exogenous INK4a and ARF caused significantly growth inhibition of A549. By FCM, more percentage of A549-p16INK4a-p14ARF cells couldn't pass through the checkpoint G1. The percentage of A549-p16INK4a-p14ARF cells inhibited at G0/G1 was 59.9%, 50.3% for A549-vector and 51.2% for A549. The statistical differences were significant between A549-p16INK4a-p14ARF cell and A549-vector cell (P=0.025) and between A549-p16INK4a-p14ARF cell and A549 cell (P=0.043). The apoptosis index of A549-p16INK4a-p14ARF cell was 8.0% and 2.7% for both A549-vector and A549 cell (P < 0.01). The colony formation ability of A549-p16INK4a-p14ARF was weaker than that of A549-vector and A549, they were 63%, 87% and 85% respectively.

CONCLUSIONSThe wild-type INK4a/ARF gene can be co-introduced effectively into A549 cell by cationic liposome method. The reexpression of p16INK4a and p14ARF in A549 can inhibit the growth and enhance the apoptosis. This trial will be helpful in using gene therapy of lung cancer in the future.

N1-methyladenosine(m1A)RNA methylation is critical for regulating mRNA translation;however,its role in the development,progression,and immunotherapy response of head and neck squamous cell carcinoma(HNSCC)remains largely unknown.Using Tgfbr1 and Pten conditional knockout(2cKO)mice,we found the neoplastic transformation of oral mucosa was accompanied by increased m1A modification levels.Analysis of m1A-associated genes identified TRMT61A as a key m1A writer linked to cancer progression and poor prognosis.Mechanistically,TRMT61A-mediated tRNA-m1A modification promotes MYC protein synthesis,upregulating programmed death-ligand 1(PD-L1)expression.Moreover,m1A modification levels were also elevated in tumors treated with oncolytic herpes simplex virus(oHSV),contributing to reactive PD-L1 upregulation.Therapeutic m1A inhibition sustained oHSV-induced antitumor immunity and reduced tumor growth,representing a promising strategy to alleviate resistance.These findings indicate that m1A inhibition can prevent immune escape after oHSV therapy by reducing PD-L1 expression,providing a mutually reinforcing combination immunotherapy approach.

Objective:To observe the clinical efficacy of combining repeated transcranial magnetic stimulation (rTMS) at low frequency with xing nao kai qiao acupuncture in the treatment of upper limb motor dysfunction after a stroke.Methods:Forty stroke survivors with upper limb motor dysfunction were randomly divided into a control group and a combination group, each of 20. All received basic neurological medication, conventional rehabilitation treatment, and xing nao kai qiao acupuncture. The combination group additionally received rTMS. Both groups were treated once a day, 5 days a week for 2 weeks consecutively. Before and after the treatment, both groups were evaluated using the Brunnstrom motor function stages, modified Ashworth spasticity grades, Fugl-Meyer upper limb motor function (FMA-UE) scoring, the modified Barthel index (MBI), and motor evoked potential (MEP) latency and amplitude.Results:The upper-limb and hand motor functioning of both groups improved significantly compared with before the treatment. The hand motor function staging of the combination group was then significantly better than among the control group. The average FMA-UE and MBI scores of both groups improved significantly, with significantly greater improvement in the combination group than in the control group. MEP latency and amplitude also improved significantly in both groups, with the average MEP latency and amplitude of the combination group superior to the control group′s averages.Conclusion:Supplementing xing nao kai qiao acupuncture with low-frequency rTMS can significantly improve the motor functioning, spasticity, and skill in the activities of daily living of stroke survivors with upper limb motor dysfunction.