IMpower210:A phase Ⅲ study of second-line atezolizumab vs. docetaxel in East Asian patients with non-small cell lung cancer

Objective: IMpower210(NCT02813785) explored the efficacy and safety of single-agent atezolizumab vs.docetaxel as second-line treatment for advanced non-small cell lung cancer(NSCLC) in East Asian patients.Methods: Key eligibility criteria for this phase Ⅲ, open-label, randomized study included age ≥18 years;histologically documented advanced NSCLC per the Union for International Cancer Control/American Joint Committee on Cancer staging system(7th edition);Eastern Cooperative Oncology Group performance status of 0 or 1;and disease progression following platinum-based chemotherapy for advanced or metastatic NSCLC. Patients were randomized 2:1 to receive either atezolizumab(1,200 mg) or docetaxel(75 mg/m^(2)). The primary study endpoint was overall survival(OS) in the intention-to-treat(ITT) population with wild-type epidermal growth factor receptor expression(ITT EGFR-WT) and in the overall ITT population.Results: Median OS in the ITT EGFR-WT population(n=467) was 12.3 [95% confidence interval(95% CI),10.3-13.8] months in the atezolizumab arm(n=312) and 9.9(95% CI, 7.8-13.9) months in the docetaxel arm[n=155;stratified hazard ratio(HR), 0.82;95% CI, 0.66-1.03]. Median OS in the overall ITT population was 12.5(95% CI, 10.8-13.8) months with atezolizumab treatment and 11.1(95% CI, 8.4-14.2) months(n=377) with docetaxel treatment(n=188;stratified HR, 0.87;95% CI, 0.71-1.08). Grade 3/4 treatment-related adverse events(TRAEs) occurred in 18.4% of patients in the atezolizumab arm and 50.0% of patients in the docetaxel arm.Conclusions: IMpower210 did not meet its primary efficacy endpoint of OS in the ITT EGFR-WT or overall ITT populations. Atezolizumab was comparatively more tolerable than docetaxel, with a lower incidence of grade3/4 TRAEs.

Modulatory effect of D-pinitol on bleomycin-induced pulmonary fibrosis in rats

Objective:To assess the effect of D-pinitol on pulmonary fibrosis induced by bleomycin.Methods:Sprague-Dawley rats received intratracheal bleomycin(6 IU/kg)to induce pulmonary fibrosis,followed by administration of either D-pinitol(5,10,or 20 mg/kg)or vehicle or methylprednisolone(10 mg/kg)over 28 days after bleomycin administration.Lung function,biochemical parameters,serum biochemistry,mRNA expressions,and histological features were observed.Results:D-pinitol at 10 and 20 mg/kg significantly(P<0.05)attenuated bleomycin-induced bronchoalveolar lavage fluid,decreased myeloperoxidase,nitric oxide,malondialdehyde levels,and increased glutathione and superoxide dismutase level.D-pinitol also improved lung function(enhanced pause,frequency of breathing,expired volume,and tidal volume).Besides,D-pinitol significantly(P<0.05)upregulated Nrf2 and downregulated mRNA expressions of TGF-β,collagen-1,and Smad-3.Furthermore,considerably less inflammation(peribronchial,perivascular,and total),Ashcroft,and interstitial fibrosis scores were observed in the D-pinitol group.Conclusions:D-pinitol exerts its effect against bleomycin-induced pulmonary fibrosis via antioxidative and anti-fibrotic pathways.

Evaluation of serum and pleural levels of endostatin and vascular epithelial growth factor in lung cancer patients with pleural effusion

Objective:To evaluate the diagnostic value of endostatin(ES),vascular endothelial growth factor (VEGF) and careinoembryonie antigen(CEA) in both serum and pleural effusion of lung cancer patients.Methods:Levels of ES,VEGF and CEA in 52 malignant pleural effusion due to lung cancer and 50 patients with non-malignant disease were measured by using sandwich enzymelinked immunosorbent assay and microparticle enzyme immunoassay.Results:The ES.VEGF and CEA levels in pleural effusion and serum,and their ratio(F/S) were higher in lung cancer group than that in benign group,and the differences were statistically significant(P【0.05).The diagnostic efficiency of ES+VEGF for lung cancer was superior to either single detection.The diagnostic efficiency of ES+VEGK+CEA was superior to either ES+VEGF or ES+CEA.Conclusions: The results suggest that ES,VEGF and CEA might be useful in the differentiation between benign and malignant pleural effusion due to lung cancer.In comparison with either single determination of concentration in serum or pleural fluid,the couiljined detection of two or three markers is of important clinical significance in the diagnosis of lung cancer.

Regulating effect of glycyrrhetinic acid on bronchial asthma smooth muscle proliferation and apoptosis as well as inflammatory factor expression through ERK1/2 signaling pathway

Objective: To study the influence of glycyrrhetinic acid(GA) on bronchial asthma(BA)smooth muscle proliferation and apoptosis as well as inflammatory factor expression and its molecular mechanism.Methods: Male SD guinea pigs were selected and made into asthma models, bronchial asthma smooth muscle cells were cultured and divided into BA group, GA group and GA + LM group that were treated with serum-free RPMI1640 culture medium, serumfree RPMI1640 culture medium containing 50 ng/mL glycyrrhetinic acid, serum-free RPMI1640 culture medium containing 50 ng/mL glycyrrhetinic acid and 100 ng/mL LM22B-10 respectively; normal guinea pigs were collected and bronchial smooth muscle cells were cultured as control group. The cell proliferation activity as well as the expression of proliferation and apoptosis genes, inflammatory factors and p-ERK1/2 was determined.Results: Proliferation activity value and m RNA expression of Bcl-2, TNF-α, IL-4, IL-6,YKL-40, protein expression of p-ERK1/2 of airway smooth muscle cell in BA group were significantly higher than those of control group while m RNA expression levels of Bax,caspase-9 as well as caspase-3 were significantly lower than that of control group(P < 0.05); proliferation activity value and m RNA expression of Bcl-2, TNF-α, IL-4, IL-6, YKL-40, protein expression of p-ERK1/2 of airway smooth muscle cell in GA group were significantly lower than those of BA group(P < 0.05) while the m RNA expression levels of Bax, caspase-9 as well as caspase-3 were significantly higher than those of BA group(P < 0.05); proliferation activity value and m RNA expression of Bcl-2, TNF-α, IL-4, IL-6, YKL-40 of airway smooth muscle cell in GA + LM group were significantly higher than those of GA group(P < 0.05) while m RNA expression levels of Bax, caspase-9 as well as caspase-3 were significantly lower that of GA group(P < 0.05).Conclusion: GA can inhibit the proliferation of bronchial smooth muscle cells and reduce the expression of inflammatory factors by inhibiting the phosphorylation of ERK1/2.

Activity of Matrix Metalloproteinase in Airway Epithelial Cells of COPD Patients

Summary:To examine the mRNA expression of matrix metalloproteinase 9 (MMP-9) and the gelatinase activity of its inhibitor, tissue inhibitor of matrix metalloproteinase 1 (TIMP-1) in the primary epithelial cells of patients with COPD, airway epithelial cells were taken from 15 COPD patients and cultured in vitro. The patients were divided into three groups, COPD group, normal smoking control group and non-smoking control group, with 5 subjects in each group, on basis of the smoking history and lung function. The semi-qualitative RT-PCR was employed to determine the mRNA levels of MMP-9 and TIMP-1 and SDS PAGE was used for the determination of the gelatinase activity of MMP-9 and TIMP-1. Our result showed that the mRNA of MMP-9 and TIMP-1 in epithelial cells of the non-smoking subjects was at a low level. The mRNA of MMP-9 and TIMP-1 in COPD patients and smokers was significantly higher than that in non-smokers (P<0.05). No significant difference was found in the levels of MMP-9 and TIMP-1 in epithelial cells between the COPD patients and smokers. The MMP-9/TIMP-1 ratios in COPD patients and smokers were significantly lower than that of non-smokers (P<0.05). The gelatinase activity in the epithelial cells of both COPD patients and normal smokers was increased (P<0.05), but no difference existed in the gelatinase activity in the epithelial cells between COPD patients and normal smokers. It is concluded that the transcription of MMP-9 and TIMP-1 and the gelatinase activity of MMP-9 and MMP-2 in the epithelial cells in COPD patients were increased, which resulted in an imbalance of MMP-9/TIMP-1, thereby causing pulmonary fibrosis. These factors play important roles in the pathogenesis of COPD.

An investigation of the effects of dust storms on rat lung using HRCT and blood gas analysis

The increasing intensity and frequency of sand-dust storms in China has led to greater prominence of associated environmentaland health issues. Many studies have focused on the health effects of air particulate contaminants, but fewformal investigations have studied the effects of sand-dust storms on human and animal health. The aim of this study wasto investigate the effects of dust storms on rat lung by using high resolution computed tomography （HRCT） and blood gasanalysis through a wind tunnel simulating. We found that the rat lung damage effects can be detected by the HRCT imagingafter exposure to sand-dust storm environments, but had no obvious result through blood gas analysis. Exposure durationspositively correlated with the damage degree to lung tissue. These will provide some evidence for clinical diagnosis ofnon-occupational pneumoconiosis.

A rare occurrence of a hereditary Birt-Hogg-Dubésyndrome:A case report

BACKGROUND Birt-Hogg-Dubé(BHD)syndrome is a rare autosomal dominant disease caused by germline mutations in the folliculin(FLCN)protein gene,which usually manifests as cutaneous fibrofolliculoma,pulmonary cysts,renal cell carcinoma,and spontaneous pneumothorax.CASE SUMMARY A 26-year-old woman with no history of smoking was admitted to the Respiratory Department of our hospital due to intermittent wheezing that lasted for 8 mo.She had experienced recurrent spontaneous pneumothorax more than four times during the past 8 mo.After admission,the patient again suffered from left pneumothorax without a clear reason.Lung computed tomography(CT)showed multiple low-density cystic changes in both lungs.Physical examination on admission revealed multiple white dome-shaped papules in the neck,the nape,and behind the ear.In addition,the patient had a family history of spontaneous pneumothorax.Her mother had suffered from pneumothorax four times(at age 36,37,42,and 50 years).Her second maternal aunt had suffered from a right pneumothorax at the age of 40.The multidisciplinary diagnosis of BHD,which included the Respiratory Department,Radiology Department,Pathology Department,and Dermatological Department,was BHD and was later confirmed by family genetic testing.The same variation(FLCN gene)was found in the patient’s mother and aunt.CONCLUSION This case highlights the importance of multidisciplinary diagnosis and a treatment platform for the diagnosis of BHD.

The expression and significance of the multidrug resistance-related proteins P-gp, MRP and LRP in human non-small cell lung cancer tissues

Objective: To explore the expression and significance of the multidrug resistance-related proteins P-glycopro-tein (P-gp), multidrug resistance-related protein (MRP), lung resistance protein (LRP) in human non-small cell lung cancer (NSCLC) tissues and paratumor tissues. Methods: Immunohistochemistry (IHC) was used to examine the expression level of proteins P-gp, MRP and LRP in 43 samples of NSCLC and 15 samples of paratumor tissues. Results: The expression rates of P-gp, MRP and LRP in 43 tumor tissues were 74.42% (32/43), 67.44% (29/43) and 88.37% (38/43), respectively, while in 15 paratumor tissues were 13.33% (2/15), 20.00% (3/15) and 6.67% (1/15), respectively. There was significant difference in the expression of proteins (P-gp, MRP and LRP) between lung cancer tissues and paratumor tissues (P < 0.05). The expres-sion of proteins P-gp, LRP in lung adenocarcinoma were higher than that in other pathological carcinomas (P < 0.05). The expression of protein MRP was not related to pathological type, clinical stage and classification of histodifferentiation (P > 0.05). Conclusion: Multidrug resistance is more common in NSCLC. The proteins of P-gp, MRP and LRP participated in the formation of multidrug resistance in lung cancer. Detection of multidrug resistance-related proteins in lung cancer tissues may be useful to choice drugs.

Abnormal expression of VEGF and its gene transcription status as diagnostic indicators in patients with non-small cell lung cancer

Objective Angiogenesis is known to be essential for the survival,growth,invasion,and metastasis of lung cancer cells. Vascular endothelial growth factor(VEGF) is an important factor regulating angiogenesis of non-small cell lung cancer(NSCLC); however,its pathologic features and significance are unclear. In this study,the tissue VEGF expression levels and its gene transcriptional status,as well as circulating VEGF levels,were investigated in patients with lung disease. Methods VEGF protein and m RNA expression levels in 38 lung tissue samples were analyzed by immunohistochemistry and reverse transcription-polymerase chain reaction(RT-PCR),respectively. Circulating VEGF levels were detected quantitatively by an enzyme linked immuno-sorbent assay. Results The level of VEGF expression was significantly higher in lung cancer tissue than in the corresponding paracancerous or non-cancerous tissues. The average level of VEGF-positive staining was 76% in tissue samples from NSCLC patients; the levels were 89% in tissue samples from stage III patients and 92% in stage IV patients. High VEGF expression was also evident in cases with lymph node metastasis(84%),distant metastasis(90%),and lower differentiation degree(89%). VEGF m RNA in cancerous tissues was represented predominantly by the VEGF121 and VEGF165 isoforms. Circulating VEGF levels were significantly higher in NSCLC patients [(840 ± 324) pg/m L] than in patients with benign lung diseases [(308 ± 96) pg/m L] or in healthy individuals serving as controls [(252 ± 108) pg/m L]. Conclusion The over-expression of lung VEGF and its gene transcription status should be useful molecular indicators for NSCLC diagnosis.

Application of TREM-1 in peripheral blood as a switching point for sequential ventilation of patients with chronic obstructive pulmonary disease complicated with respiratory failure

Objective:To investigate the application of peripheral blood triggering receptor expressed on myeloid cells-1(TREM-1)for sequential treatment switching points in patients with chronic obstructive pulmonary disease(COPD)complicated with respiratory failure.Methods:A total of 120 cases of COPD patients with respiratory failure from June 2017 to December 2018 were randomly divided into two groups:60 cases in the control group and 60 cases in the observation group.The control group received spontaneous breathing trials for 2 h to select the time for non-invasive positive pressure ventilation,while the observation group received peripheral blood TREM-1(≤90.0 pg/mL)to select the time for non-invasive positive pressure ventilation.The stress hormones,clinical pulmonary infection score and vital signs of two groups after 24 h of mechanical ventilation were detected.The treatment time and the adverse reactions of the two groups were recorded.Results:There was no significant difference in rennin,adrenaline,noradrenaline and angiotensin II between two groups(P>0.05).Compared with the control group,the clinical pulmonary infection score was decreased in the observation group(P<0.05).There was no significant difference in heart rate,respiratory rate,pH,partial pressure of carbon dioxide and partial pressure of oxygen between two groups(P>0.05).There was no significant difference in intensive care monitoring time between two groups(P>0.05).Compared with the control group,the observation group had no significant difference in invasive ventilation time and total mechanical ventilation.The time of hospitalization and hospitalization had significantly decreased(P<0.05).There was no significant difference in mortality,ventilator-associated pneumonia and re-intubation between two groups(P>0.05).Conclusion:TREM-1 can be used as a switching point during invasive-noninvasive sequential ventilation for COPD patients with respiratory failure,which can shorten the time of invasive ventilation,total mechanical ventilation and hospitalization.

Is the traditional Chinese medicine helpful for patients with hematologic malignant diseases? A meta-analysis of randomized controlled trials

AIM:To evaluate the efficacy of traditional Chinese medicine(TCM)for the treatment of hematologic malignant diseases.METHODS:We searched the Cochrane CENTRAL,PubM ed,Embase,Web of Science,AMED,CNKI,Wanfang Platform;China Sinomed and the clinical trial registry web sites and Googlescholar electronically up to June19th,2014 and hand searched related publications.Only randomized controlled trials(RCTs)researching on whether TCM as the adjuvant treatment improved the effect for hematologic malignant diseases were included.Two reviewers extracted data and evaluated the studies independently.Pooled risk ratios(RR)were calculated as outcome measures.Our primary outcomes were the overall response(OR)rate.RESULTS:We retrieved 13143 references and included11 RCTs involved 891 participants after screening.Because the non-significant heterogeneity we used the fixed effect model to combine data and TCM had a significantly higher OR and CR(complete response)rates than the control[RR=1.17,95%CI:(1.10,1.25),P<0.00001;RR=1.24,95%CI:(1.11,1.37),P<0.0001,respectively].Only three studies included in the survival rate analysis.We combined them with random effects model and there was no significant difference between the TCM and control arms.Becauseof the low heterogeneity we used the fixed effect model to combine the non-hematologic adverse effects(AEs)data.Our results showed that TCM significantly decreased non-hematologic AEs rates we researched,the gastrointestinal reaction[RR=0.50,95%CI:(0.37,0.68),P<0.0001],liver and/or kidney injury[RR=0.37,95%CI:(0.26,0.53),P<0.00001]and heart injury[RR=0.24,95%CI:(0.09,0.68),P=0.007].Additionally,TCM had a trend to decrease the infection rate[RR=0.16,(0.02,1.12),P=0.07],but not statistically significantly.CONCLUSION:TCM increases OR and CR rates for hematologic malignances and reduces treatment associated serious non-hematologic AEs.Therefore,TCM should be included in the treatment of hematologic malignances.

miR-3622b-5p regulates cisplatin resistance of human gastric cancer cell line by targeting BIRC5

Many evidences showed that drug resistance of gastric cancer cells could be regulated by the abnormal expression of microRNAs(miRNAs),a post-transcriptional regulator of gene expression.Thus,we investigated the role of miR-3622b-5p in the development of cisplatin resistance in human gastric cancer cell lines.A set of biochemical assays were used to elucidate the mechanism by which miR-3622b-5p regulates drug resistance in cancer cells.The expression of miR-3622b-5p was measured by quantitative real-time PCR and showed that MiR-3622b-5p was significantly downregulated in the plasma of patients with acquired drug resistance to platinumbased chemotherapy for gastric cancer.MiR-3622b-5p was also found significantly downregulated in cisplatinresistant gastric cancer cell line SGC7901/cisplatin(DDP),compared with the parental SGC7901 cells.An in vitro drug sensitivity assay showed that overexpression of miR-3622b-5p sensitized SGC7901/DDP cells to cisplatin.The luciferase activity of reporters constructed by BIRC53′-untranslated regions in SGC7901/DDP cells suggested that BIRC5 was target gene of miR-3622b-5p.Ecpotic miR-3622b-5p expression in SGC7901/DDP cells significantly repressed the expression of the BIRC5 and sensitized the cells to DDP-induced apoptosis.By contrast,treatment with miR-3622b-5p inhibitor increased the protein expression of BIRC5 and led to a lower proportion of apoptotic cells in the SGC7901 cells.In conclusion,our findings suggest that miR-3622b-5p regulates cisplatin resistance of human gastric cancer cells at least in part by repressing the expression of BIRC5.Altering miR-3622b-5p expression may be a potential therapeutic strategy for the treatment of chemoresistance in GC in the future.

An Outbreak of SARS in a Diabetes Room of a General Hospital without Infected Medical Staff

Objective To investigate the epidemiologic features of an outbreak of SARS that occurred in a single diabetes room of a general hospital in Beijing in late March 2003.Methods Field investigation was carried out in the ward,the nursing log and the hospitalization medical record of correlative patients were consulted.SARS-CoV in serum specimen from SARS patient was detected by PCR.Results The room where SARS outbreak occurred was on the 13th floor of the 16-story main ward building.There were 6 beds in the room,living with 6 female patients(aged 45-67)who were all hospitalized due to type2 diabetes.On March 24,2003,Patient 1 began to have a fever and cough,chest X-ray showed pneumonia.Five and six days later,Patient 2 and Patient 3 began to have a fever,respectively.Finally,all of these 3 patients died.Their beds were all at the same side of the room,and the other 3 patients at the opposite side were not infected.Serum SARS CoV-RNA of the Patient 3 was positive by nest-PCR.The daughter-in-law of Patient 1who accompanied Patient 1 by the bedside several days,mainly near the window,upwind of Patient 1,was not infected.Medical staff,family members and visitors of the 6 patients were not infected.Conclusions This outbreak was not transmitted by aerosol.The distance droplets travels could be up to 3.43 meters.Droplet spread has direction,and the droplets direction of propagation is closely related with the wind direction and speed.Those at the downwind position of SARS patients were susceptible to be infected.Medical staff wore face masks and good natural ventilation of this ward building may be important reasons for the prevention of infection.

The relationship between diabetes‑related distress and self‑management in empty‑nest elderly patients with type 2 diabetes mellitus:The mediating effect of self‑efficacy

Objective:The aim of this study is to investigate the mediating effect of self‑efficacy between diabetes‑related distress and self‑management in empty‑nest elderly patients with type 2 diabetes mellitus(T2DM).Methods:A total of 347 empty‑nest elderly patients with T2DM from Jiangsu Province Hospital of Chinese Medicine and Affiliated Hospital of Yangzhou University were investigated by a self‑designed General Information Questionnaire,the Self‑efficacy for diabetes,the Diabetes Distress Scale,and the summary of diabetes self‑care activities.Diabetes‑related distress and diabetes self‑management were tested for correlation analysis with self‑efficacy.Meditational analyses were conducted to test the hypothesized mediating effects of diabetes self‑efficacy as an intervention variable between diabetes distress and self‑management.Results:About 68.59%of patients suffered from diabetes‑related distress.The results of bivariate correlations revealed that self‑efficacy was negatively correlated with diabetes‑related distress(r=–0.627,P<0.01)and positively correlated with self‑management(r=0.356,P<0.01).The relationship effects of diabetes distress on self‑management were fully mediated by self‑efficacy.Conclusions:Diabetes‑related distress prevails in empty‑nest elderly patients with T2DM.Self‑efficacy plays a mediate role between diabetes‑related distress and self‑management.Psychological intervention is contributed to reducing diabetes‑related distress.

Cloning and screening of cDNA of Psilgramma menephorn allergen

Objective To construct a cDNA expression library of Psilgramma menephorn to screen its major allergen so as to provide the basis for producing recombinant allergen vaccine of Psilgramma menephorn. Methods Total RNA was extracted from the whole body of Psilgramma menephorn with Trizol and mRNA was purified with Oligo (dT) Spin-Column. And dscDNA was synthesized through reverse transcription. After blunting, the cDNA fragments were ligated with EcoRⅠ adapters. Then the cDNAs were digested by XhoⅠ, and the fragments less than 400 bp were removed by using GHROMA SPIN-400 column. The remaining fragments longer than 400 bp were ligated with Uni-ZAP XR vector. The recombinants were packaged in vitro and a small portion of the packaged phage was used to infect E.coli XL1-Blue MRF′ for titration. The recombinants were examined by color selection. The size of cDNA inserts and the diversity of library were analyzed by PCR. The library was screened using SPT positive sera from patients with Psilgramma menephorn allergy repeatedly. Results The cDNA expression library consisting of a 5×105 recombinant bacteriophages was constructed with the recombinant ratio of 67%. The average length of recombinant exogenous inserts was about 1.49 kb. Five positive cDNA clones were obtained. Conclusion The constructed cDNA expression library shows appropriate contents and size of cDNA fragments and the related genes of Psilgramma menephorn major allergens were harbored successfully, which lays the foundation for the positive clone identification and further analysis.

THE CLINICAL EVALUATION OF ANGIOGENESIS INHIBITOR-20(R)-GINSENOSIDE RG_3 IN LUNG CANCER

Objective The aim of the study was to make a further evaluation of Ginsenoside Rg3. Methods The clinical effects of the drug on moderate and advanced lung cancer, including side effects, were observed. Results Ginsenoside Rg3 improved chemotherapy significantly. The clinical relief rate of patients treated with antiangiogenic agent 20 (R) Ginsenoside Rg3 was 36.6%, which was higher than that of the patients not treated with it (16.7%)( P <0.05). It had no significantly different effects on lung cancers of different types of tissues ( P >0.05). It provided better treatment on the cancer at early stage than that at advanced stage ( P <0.05). Moreover the living qualities of the patients were improved notably ( P <0.05). Conclusion Combined with chemotherapy, angiogenesis inhibitor 20(R) Ginsenoside Rg3 can improve clinical therapeutic efficacy and the living qualities of patients significantly.

Construction, Expression and Characterization of a Chimeric Protein Targeting Carcinoembryonic Antigen in Lung Cancer

The carcinoembryonic antigen（CEA） is an oncofetal glycoprotein known as an important clinical tumor marker and is overexpressed in several types of tumors, including colorectal and lung carcinomas. We constructed a chimeric protein that exhibits both specific binding and immune stimulating activities, by fusing staphylococcal enterotoxin A（SEA） to the C-terminus of an anti-CEA single-chain disulfide-stabilized Fv（scdsFv） antibody （single-chain-C-terminus/SEA, SC-C/SEA）. The SC-C/SEA protein was expressed in Escherichia coli（E. coli）, refolded, and purified on an immobilized Ni2＋ affinity chromatography column. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis（SDS-PAGE） and Western blot analysis reveal that the target protein was expressed sufficiently. We used immunofluorescence assays to demonstrate that SC-C/SEA could bind specifically to human lung carcinoma cells（A549）, but almost human uterine cervix cells（HeLa）. We also used the L-lactate dehydrogenase（LDH） release assay to show that SC-C/SEA elicits a strong A549 tumor-specific cytotoxic T lymphocyte（CTL） response in vitro. The results suggest that SC-C/SEA shows specific activity against CEA-positive cells and has potential application in CEA-targeted cancer immunotherapy.

Clinicopathologic significance of CXCR4 and Nrf2 in colorectal cancer

The CXCR4 and Nrf2 signaling pathways are abnormally activated in response to cellular stress in various types of human cancers. In this study, we examined the expression of CXCR4 and Nrf2 in colorectal cancer （CRC） tissue specimens and investigated their correlation with patient clinicopathologic characteristics. We determined CXCR4 and Nrf2 expression in 76 CRC tissue specimens and paired normal tissue specimens by immunohistochemistry and real-time PCR. We found that the protein and mRNA transcript levels of CXCR4 were significantly higher in CRC tissue specimens than in paired normal tissues, while the expressions of Nrf2 protein and mRNA were increased in CRC tissues compared to distant non-cancerous tissues. High expression level of CXCR4 was positively correlated with poorly differentiated （P=0.031）, more advanced tumor-node-metastasis （TNM） stage （P=0.019）, lymph node metastasis （P=0.007） and distant metastasis （P=0.018）. However, the expression of Nrf2 protein was positively correlated with larger tumor size （P=0.049）, more advanced TNM stage （P=0.013）, lymph node metastasis （P=0.016） and distant metastasis （P=0.023）. Moreover, there was a strong relationship between CXCR4 and Nrf2 expression in CRC tissues, indicating that high Nrf2 expression may contribute to CXCR4 overexpression. In addition, combined expression of CXCR4 and Nrf2 strongly correlated with lymph node metastasis and distant metastasis （P=0.003）. Furthermore, we found that combined high expression of CXCR4 and Nrf2 had stronger correlation with lymph node metastasis and distant metastasis than any single molecule did. This study indicated that the abnormal expression of CXCR4 and Nrf2 contributed to the progression of CRC.

Effect of grape proanthocyanidins on tumor growth and angiogenesis in H22 liver cancer xenograft model

Objective: The aim of this study was to investigate the effect of grape proanthocyanidins(GPC) on the growth and angiogenesis of hepatocellular carcinoma H22 cells xenograft in mice. Methods: The xenograft model was established using injected subcutaneously H22 cells into the right axilla of the mice. Each group was treated with different doses of GPC and Endostar. All these treatments were maintained for 10 days, and mice were sacrificed. The xenograft tumors in mice were measured. The proliferation activity level of H22 cells was determined by MTT assay, and the levels of vascular endothelial growth factor(VEGF) protein were examined by immunohistochemistry. Results: When treated with 50, 100 and 200 mg/kg of GPC and Endostar, the tumor inhibition rates were 13.17%, 23.37%, 36.15% and 14.71%, respectively. The tumor weight of xenograft was significantly lighter in high GPC group than the control group(P < 0.05). The ODs in GPC groups were 0.835, 0.666 and 0.519, respectively. The absorbances in middle and high GPC groups were statistically significant, compared with control group(P < 0.01). Immunohistochemical technique showed the expression of VEGF of the GPC groups was downregulated significantly compared with the control group(P < 0.01). Conclusion: GPC can inhibit the growth of hepatocellular carcinoma H22 cell xenograft in mice. The inhibition of angiogenesis by the down-regulation of VEGF expression may play a key role in the anti-neoplastic effect of GPC.

Thymidylate synthase confers pemetrexed resistance of non-small cell lung cancer cells by EGFR/PI3K/AKT pathway

Chemotherapy drug resistance is the main cause leading to the relapse and metastasis of non-small cell lung cancer(NSCLC)patients.Our study aimed to investigate the mechanism of pemetrexed resistance in NSCLC.Firstly,the pemetrexed(PEM)-resistant PC-9 and A549 lung adenocarcinoma cell lines(PC-9/PEM and A549/PEM)were established.The expression of thymidylate synthase(TS)in PC-9/PEM,A549/PEM,A549,and PC-9 cells were analyzed by qRT-PCR and western blot.Then,cell viability,colony formation,migration,and invasion were performed on PEM-resistant cells transfected with TS siRNA.The role of EGFR in PEM resistance of PEM-resistant cells was investigated using EGFR siRNA.The effects of gefitinib and EGFR siRNA on EGFR/PI3K/AKT pathway and downstream signaling Cyclin D1 and E2F1 in PEM-resistant cells were analyzed.Results showed that the protein level of TS was significantly increased in A549/PEM and PC-9/PEM.TS knockdown inhibited the potency of proliferation,colony-forming potential,migration,and invasion in PEM-resistant cells.EGFR knockdown abrogated the resistance to PEM of PEM-resistant cells and suppressed the migration and invasion of PEM-resistant cells.Gefitinib treatment and EGFR knockdown respectively inhibited the EGFR/PI3K/AKT pathway and downregulated Cyclin D1 and E2F1 in PEM-resistant cells.Thus,TS might be a predictive marker for PEM resistance in NSCLC.Inhibition of the EGFR pathway abrogated the resistance to PEM and inhibited the EGFR/PI3K/AKT and downstream signaling of PEM-resistant NSCLC cell lines.