Bevacizumab Combined with Icotinib Overcomes Osimertinib Resistance in a Patient of Non-Small Cell Lung Cancer.

A 61-year-old Chinese woman was diagnosed as primary pulmonary adenocarcinoma of left superior lobe with epidermal growth factor receptor (EGFR) 19 del mutation positive. Treatment with icotinib was given, but her disease progressed after 6 months remission. CT-guide needle biopsy for the new lesion in inferior lobe of left lung demonstrated intrapulmonary metastasis, and EGFR gene panel by Amplification Refractory Mutation System Polymerase Chain Reaction (ARMS-PCR) confirmed EGFR T790M mutation. Treatment with osimertinib was initiated. After 2 months remission, the disease progressed. Re-biopsy was performed for the tumor in the inferior lobe of left lung, and ARMS-PCR demonstrated no other gene mutation except EGFR 19 del. Icotinib was re-challenged, but disease progressed continuously. Bevacizumab was added, and partial response was achieved after 2-cycle of combination therapy. The non-small cell lung cancer (NSCLC) in this case maintained EGFR activating mutation and lost EGFR T790M mutation was a genetic change after osimertinib treatment. This case suggests the re-challenge of the first-generation EGFR-TKIs combined with bevacizumab may overcome the tumor resistance and prolong survival of NSCLC patient.

[Relationship between epidermal growth factor receptor gene expression and radiosensitivity of non-small-cell lung cancer cells].

OBJECTIVE: To explore the relationship between epidermal growth factor receptor (EGFR) gene expression and radiosensitivity of non-small-cell lung cancer (NSCLC) cells. METHODS: EGFR sequence-specific double-stranded RNA (dsRNA-EGFR) was chemically synthesized. NSCLC cell line SPC-A1 was transfected with dsRNA-EGFR formulated with Lipofectamine 2000. Western blot and real-time PCR were used to determine the EGFR mRNA and protein expression, respectively. Colony inhibition test was adopted to observe the radiosensitizing effect. To establish the nude mouse tumor models, calculate the tumor growth inhibition rate and make the tumor growth curve by measuring its size and weight. RESULTS: EGFR mRNA levels were 1.51 ± 0.22, 1.38 ± 0.15 and 0.45 ± 0.11 in the control group, dsRNA-unrelated group and dsRNA-EGFR group, respectively (F = 482.7, P < 0.01). The contents of EGFR protein were 2340.87 ± 10.99, 2231.85 ± 35.66 and 832.03 ± 39.13 in the control group, dsRNA-unrelated group and dsRNA-EGFR group, respectively (F = 263.3, P < 0.05). Compared with the control group, dsRNA-EGFR sequence specifically decreased the expressions of EGFR mRNA by 70.2% and EGFR protein by 64.5%. The colony inhibition rates of the control group, dsRNA-unrelated combined with radiotherapy group and dsRNA-EGFR combined with radiotherapy group were 9.3%, 12.5% and 65.5%, and the tumor growth inhibition rates were 21.3%, 24.4% and 64.2%, respectively. The combination of dsRNA-EGFR and radiotherapy significantly inhibited the tumor growth in vitro and in vivo. CONCLUSIONS: DsRNA-EGFR shows an apparent inhibitory effect on the expression of EGFR mRNA and protein of NSCLC cells, effectively inhibit the tumor growth in vivo, and enhance the radiosensitivity of NSCLC.

[The diagnostic value of medical thoracoscopy for unexplained pleural effusion].

OBJECTIVE: To explore the endoscopic features of patients with unexplained pleural effusion, and to evaluate the diagnostic value of medical thoracoscopy. METHODS: A retrospective analysis of 2380 patients with unexplained pleural effusion (1320 males and 1060 females; age 15-94 years) in Shandong Provincial Hospital from 1992 to 2011 were performed .The diagnosis was confirmed by medical thoracoscopy. RESULTS: The endoscopic findings of malignant pleural effusion mostly showed nodules of varying sizes. The nodules could be grape-like, cauliflower-like, fused into masses, or diffused small nodules . The appearance of cancerous nodules was more diversified compared to tuberculous nodules. Tuberculous pleurisy was manifested as diffuse pleural congestion and miliary changes, multiple small gray-white nodules, fibrin deposition and adhesion in the pleural cavity, pleural thickening and loculation . The pathological diagnosis was as follows: pleural metastases in 899 (37.8%), primary pleural mesothelioma in 439 (18.4%), tuberculous pleurisy in 514 (21.6%), non-specific inflammation in 226 (9.5%), empyema in 190 (8.0%), hepatic pleural effusion in 36 (1.5%) and pleural effusion of unknown causes in 76 (3.2%) cases. The diagnostic positive rate of medical thoracoscopy was 96.8%. No serious complications were observed. CONCLUSION: Medical thoracoscopy is a relatively safe procedure and has an important application value in the diagnosis of unexplained pleural effusion.

Methodological evaluation and comparison of five urinary albumin measurements.

BACKGROUND: Microalbuminuria is an indicator of kidney damage and a risk factor for the progression kidney disease, cardiovascular disease, and so on. Therefore, accurate and precise measurement of urinary albumin is critical. However, there are no reference measurement procedures and reference materials for urinary albumin. METHODS: Nephelometry, turbidimetry, colloidal gold method, radioimmunoassay, and chemiluminescence immunoassay were performed for methodological evaluation, based on imprecision test, recovery rate, linearity, haemoglobin interference rate, and verified reference interval. Then we tested 40 urine samples from diabetic patients by each method, and compared the result between assays. RESULTS: The results indicate that nephelometry is the method with best analytical performance among the five methods, with an average intraassay coefficient of variation (CV) of 2.6%, an average interassay CV of 1.7%, a mean recovery of 99.6%, a linearity of R=1.00 from 2 to 250 mg/l, and an interference rate of <10% at haemoglobin concentrations of <1.82 g/l. The correlation (r) between assays was from 0.701 to 0.982, and the Bland-Altman plots indicated each assay provided significantly different results from each other. CONCLUSION: Nephelometry is the clinical urinary albumin method with best analytical performance in our study.

Association of the interferon-gamma single nucleotide polymorphism +874(T/A) with response to immunosuppressive therapy in patients with severe aplastic anemia.

The aim of this study was to investigate whether a single nucleotide polymorphism (SNP) +874T/A of interferon-gamma (IFN-γ) correlates with response to immunosuppressive therapy. Amplification refractory mutation system-polymerase chain reaction was used to amplify the polymorphic segments of the IFN-γ +874T/A gene in the samples obtained from 54 patients with aplastic anemia and 51 healthy adults. Further, enzyme linked immunosorbent assay was used to assay IFN-γ levels in the blood plasma of 35 patients with severe aplastic anemia before immune suppression therapy and 20 healthy blood donors. The results showed that the frequency of IFN-γ +874 TT genotype in patients with aplastic anemia was significantly higher than the corresponding frequency in the healthy adults (42.6% vs. 17.6%, χ(2)=13.78, p=0. 01). The response rate in severe aplastic anemia patients with increased IFN-γ levels in the blood plasma was higher than that in severe aplastic anemia patients with decreased IFN-γ levels in the blood plasma (73.7% vs. 25.0%, p<0.05). Of the 35 patients with severe aplastic anemia, 15 showed the IFN-γ +874 TT genotype, whereas response in 11 patients, the high response rate was significantly in the favor of the IFN-γ +874 TT genotype (73.3% vs. IFN-γ +874 non-TT 35%, p<0.05). In conclusion, the IFN-γ +874T/A gene polymorphism may be correlated with response to immunosuppressive therapy.

[Studies on the predictors of response to immunosuppressive therapy in severe aplastic anemia patients].

OBJECTIVE: To assay the factors that may predict the response to immunosuppressive therapy (IST) in severe aplastic anemia patients. METHODS: The blood samples were collected from 37 patients diagnosed as severe aplastic anemia in West China Hospital of Sichuan University and West China Second Hospital of Sichuan University during February, 2006 to March, 2007. Twenty healthy blood donors were used as normal control. The plasma levels of IFN-gamma and IL-2 were measured by enzyme linked immunosorbent assay, and the gene phenotype of HLA-DRB1 * 15 and HLA-DRB1 * 1501 was analyzed by polymerase chain reaction with sequence specific primer. The expressions of CD55 and CD59 on the cellular membrane of red blood cell and white blood cells also were measured. RESULTS: The response rate in the patients who had higher IL-2 level before IST was significantly better than that in the patients with lower IL-2 level (66.7% vs 28.6%, P<0.05), while similar result was observed to IFN-gamma (73.7% vs 25.0%, P<0.05). The response rate in the patients with positive HLA-DRB1 * 15 was higher than that in those negative patients (62.5% vs 44.4%, P>0.05), but there was no different found in different HLA-DRB1 * 1501 phenotypes (50% vs 50%, P>0.05). The response rates in the patients with deficient CD55 and CD59 expression were higher than those expressed CD55 and CD59 normally (80.0% vs 40.0%, P>0.05); the response rate of patients younger than 40 years was higher than those older than 40 years (60.0% vs 14.3%, P<0.05); the response rate in female patients was higher than male patients (62.5% vs 42.9%, P<0.05). CONCLUSION: The concentration of IL-2, IFN-gamma before IST, and age can be used as the predicting factors to immunosuppressive therapy, while the predicting value of HLA-DRB1 * 15, HLA- DRB1 * 1501 and CD55, CD59 to the response of IST still remain unclear.

[Effects of trichostatin A and paclitaxel on apoptosis and microtubule stabilization in endometrial carcinoma cells: an in vitro research].

OBJECTIVE: To investigate the effects of trichostatin A (TSA) and paclitaxel (PTX) on the apoptosis and microtubulin stabilization in human endometrial carcinoma cells and its mechanism. METHODS: Human endometrial carcinoma cells of the line Ark2, KLE and AN3 were cultured in the presence of TSA (TSA group), or PTX (PTX group), or TSA plus PTX (TSA + PTX group) respectively. The growth curve was obtained by trypan-blue exclusion assay. Apoptosis was observed by annexin V and Hoechst staining. Perturbation of mitochondrial membrane potential (MMP) was detected by flow cytometry. Western blotting was used to detect the protein expression of caspase-9, poly ADP-ribose polymerase (PARP), and acetylated microtubulin. RESULTS: The growth of the Ark2, KLE, and AN3 cells of the TSA, PTX, and TSA + PTX group, especially in the latter group, was inhibited. The Ark2 cell apoptotic rates 4 days later of the TSA, PTX, TSA + PTX, and control group were 4.25% +/- 0.25%, 12.12% +/- 0.62%, 16.56% +/- 0.74%, and 46.78% +/- 2.68% respectively by annexin V staining, and 3.39% +/- 0.12%, 6.00% +/- 0.25%, 10.05% +/- 0.53%, and 22.30% +/- 1.25% respectively by Hoechst staining. The apoptotic rates of the TSA + PTX group by both staining methods were both significantly higher than those of the other groups (all P < 0.05). Lysis of caspase-9 and PARP in the Ark2 and KLE cells increased greatly 24 hours after the TSA and PTX treatment. The disappearance rate of MMP in the Ark2 and AN3 cells of the TSA + PTX groups were 16.80% +/- 0.92% and 11.28% +/- 0.78% respectively, significantly higher than that of the PTX group (5.34% +/- 0.45% and 5.61% +/- 0.56% respectively) and TSA group (4.96% +/- 0.47% and 6.46% +/- 0.62% respectively, all P < 0.05). The expression of acetylated microtubulin was increased in the Ark2 and KLE cells of the TSA + PTX groups. CONCLUSIONS: Synergy of TSA and PTX inhibits the cell growth and induces apoptosis. The acetylation of non-histone protein induced by histone deacetylase inhibitor is one of the possible mechanisms of its anti-cancer effects.

Genetic polymorphisms of ADH2 and ALDH2 association with esophageal cancer risk in southwest China.

AIM: To evaluate the impact of alcohol dehydrogenase 2 (ADH2) and aldehyde dehydrogenase 2 (ALDH2) polymorphisms on esophageal cancer risk. METHODS: One hundred and ninety-one esophageal cancer patients and 198 healthy controls from Yanting County were enrolled in this study. ADH2 and ALDH2 genotypes were examined by polymerase-chain-reaction with the confronting-two-pair-primer (PCR-CTPP) method. Unconditional logistic regression was used to calculate the odds ratios (OR) and 95% confidence interval (95% CI). RESULTS: Both ADH2 1 allele and ALDH2 1/ 2 allele showed an increased risk of developing esophageal cancer. The adjusted OR (95% CI) for ADH2 1 allele compared with ADH2 2/ 2 was 1.65 (95% CI = 1.02-2.68) and 1.67 (95% CI = 1.02-2.72) for ALDH2 1/ 2 compared with ALDH2 1/ 1. A significant interaction between ALDH2 and drinking was detected regarding esophageal cancer risk, the OR was 1.83 (95% CI = 1.13-2.95). Furthermore, when compared with ADH2 2/ 2 and ALDH2 1/ 1 carriers, ADH2 1 and ALDH2 2 carriers showed an elevated risk of developing esophageal cancer among non-alcohol drinkers (OR = 2.46, 95% CI = 0.98-6.14), and a significantly elevated risk of developing esophageal cancer among alcohol drinkers among alcohol drinkers (OR = 9.86, 95% CI = 3.10-31.38). CONCLUSION: ADH2 and ALDH2 genotypes are associated with esophageal cancer risk. ADH2 1 allele and ALDH2 2 allele carriers have a much higher risk of developing esophageal cancer, especially among alcohol drinkers.

Risk factors for esophageal cancer: a case-control study in South-western China.

Esophageal cancer is a crucial cancer in China. Yanting in Sichuan Province was a key area with highest esophageal cancer mortality in China, but little evidence on esophageal cancer risk factors has been reported for this area and the etiology remains unclear. To clarify risk factors, a 1:1 matched case-control study was conducted. Totals of 185 eligible esophageal cancer patients and 185 healthy residents matched for sex and age were recruited. Conditional logistic regression was used to calculate odds ratios (ORs) and 95% confidence intervals (CIs) for possible risk/protective factors. All ORs were adjusted by family history of esophageal cancer and occupation, and then further adjusted by other possible confounding factors. Our results showed that smoking and alcohol drinking were risk factors for esophageal cancer with dose-response. The ORs (95% CI) compared with never smokers and drinkers were 4.06 (1.55-10.6) and 2.49 (1.06-5.85), respectively. The OR was further increased to 8.86 (95% CI, 3.82-20.5) for both smoking and drinking in combination. Eating food rapidly (OR=5.84, 95% CI, 2.05-16.7), drinking shallow ground water (OR=4.18, 95% CI, 1.30-13.4) and frequent intake of picked vegetables (OR=2.12, 95% CI, 1.00-4.49) appeared to increase the risk, while frequent intake of fresh fruit (OR=0.42, 95% CI, 0.19-0.89), fresh vegetables (OR= 0.62, 95% CI, 0.32-1.17) and eggs (OR=0.59, 95% CI, 0.25-1.39) decreased the risk. In conclusion, smoking and alcohol drinking are common in Yanting and main contributors to esophageal cancer. Consumption of fresh fruit and eggs are not common and high consumption of these two foods as well as fresh vegetables may decrease the risk of esophageal cancer in this area. In addition, drinking shallow ground water and eating food rapidly, as well as frequent intake of pickled vegetables, are also factors increasing the risk.

Imaging study on the optic canal using sixty four-slice spiral computed tomography.

BACKGROUND: Rapid advances in multislice computed tomography (MSCT) technology facilitate accurate clinical imaging. The newly developed 64-slice CT increases temporal and spatial resolution efficiently. PURPOSE: The purpose of this study is to evaluate the application of 64 slice spiral computed tomography (CT) on the imaging of the normal optics canal. METHODS AND MATERIALS: 100 healthy adults were investigated using 64 slice spiral CT. The optics canal was scanned, reconstructed and examined. RESULTS: Among the four walls of the optic canal, the medial wall is the longest one. The upper wall and outer wall are inferior to the medial wall while the inferior wall is the shortest one. All the data accomplished by the 64 slice CT was consistent with the results of previous reports using other methods. CONCLUSION: The results suggested that the 64 slice spiral CT could be a valuable and accurate method for measuring the length of optics canal walls.

[Effect and mechanism of arsenic trioxide on chemosensitivity of human lung adenocarcinoma cells].

OBJECTIVE: To study the apoptosis and chemosensitive effect of As(2)O(3) on human lung adenocarcinoma cell line A549. METHODS: MTT colorimetric assay, immunohistochemical method, flow cytometry and RT-PCR were applied to observe the different effects induced by As(2)O(3) with different concentrations on A549 cells. RESULTS: 1, 2 micro mol/L As(2)O(3) increased Fas expression and inhibited the expression of bcl-2, MRP and LRP; G(1) arrest was observed and the cells were more sensitive to cisplatin. A549 cells in the presence of 5 micro mol/L As(2)O(3) showed growth inhibition and up-regulation of bcl-2, MRP and LRP expressions; but their chemosensitivity to cisplatin did not change. CONCLUSION: As(2)O(3) in low concentrations can increase the chemosensitivity of A549 cells by regulating the expression of apoptosis genes, anti-apoptosis genes and drug-resistant genes and affecting the cell cycle progress.

[The relationship between transforming growth factor-beta1, platelet derived growth factor and airway remodeling in eosinophilic bronchitis].

OBJECTIVE: To study the relationship between the expression of transforming growth factor-beta(1) (TGF-beta(1)) and platelet derived growth factor (PDGF) and airway remodeling in eosinophilic bronchitis (EB). METHODS: Bronchial biopsy specimens were obtained from 12 patients with EB (A group), 10 asthmatic patients (B group) and 10 patients (C group) with peripheral lung cancer in early stage. The subepithelial basement membrane (SBM) thickness was measured by light microscopy using HE staining. The expressions of TGF-beta(1) and PDGF in the bronchial mucosa were examined by immunostaining. RESULTS: The SBM of A group [(6.3 +/- 1.4) micro m] was significantly thicker than that of C group [(4.1 +/- 1.2) micro m, P < 0.05], but significantly thinner than that of B group [(8.2 +/- 1.5) micro m]. The numbers of positive cells for TGF-beta(1) and PDGF in A group (59 +/- 9, 47 +/- 7 respectively) and B group (85 +/- 12, 76 +/- 11, respectively) were significantly higher than those in C group (31 +/- 4, 20 +/- 3, respectively), and were positively correlated with SBM thickness (r = 0.76, 0.52, P < 0.05). CONCLUSION: These results suggest that TGF-beta(1) and PDGF expressions in bronchial mucosa may play a role in bronchial subepithelial fibrosis in EB patients.

The effects of PTBP3 silencing on the proliferation and differentiation of MKN45 human gastric cancer cells.

AIMS: PTBP3 overexpression inhibits the differentiation of leukemia cells; however, its effects on the differentiation and proliferation of solid cancer cells remain unclear. Thus, the impact of PTBP3 on the differentiation and proliferation of gastric cancer cells was investigated. MAIN METHODS: PTBP3 expression was analyzed in normal and tumor tissues using immunohistochemistry. A xenograft model was established in nude mice by subcutaneous injection of untransfected human gastric cancer MKN45 cells or those expressing a control vector or PTBP3 siRNA. We analyzed the tumor inhibition rate, the expression of PTBP3, the PCNA-positive rate and the serum levels of CEA, CA199, CA125, LDH, ALP and γ-GT in different groups. KEY FINDINGS: The tumor weights in the PTBP3 siRNA group were significantly lower than that of the MKN45 cell control group (P<0.001). Immunohistochemistry analysis of PCNA expression revealed that it was markedly reduced after PTBP3 silencing. ELISAs showed that the serum levels of CEA and CA199 tumor markers as well as LDH and ALP were reduced after PTBP3 silencing. Transmission electron microscopy revealed that MKN45 cells expressing PTBP3 siRNA had reduced nuclear-to-cytoplasmic ratio and regular nuclei, suggesting differentiation. SIGNIFICANCE: PTBP3 may promote proliferation and inhibit the differentiation of human gastric cancer MKN45 cells.

Simultaneous blockage of epidermal growth factor receptor and cyclooxygenase-2 in a human xenotransplanted lung cancer model.

The effects of erlotinib combined with celecoxib in a lung cancer xenograft model were here explored with a focus on possible mechanisms. A xenotransplanted lung cancer model was established in nude mice using the human lung cancer cell A549 cell line and animals demonstrating tumour growth were randomly divided into four groups: control, erlotinib, celecoxib and combined (erotinib and celecoxib). The tumor major axis and short diameter were measured twice a week and after 40 days tissues were collected for immunohistochemical analyses of Bcl-2 and Bax positive cells and Western-blotting analyses for the epidermal growth factor recepto (EGFR), P-EGFR, and cyclooxygenase-2 (COX-2). Tumor size in the combined group was smaller than in the others (p<0.01) and the percentage of Bcl-2 positive cells was fewer in most cases (p<0.01), while that of Bax positive cells was greater than in the erlotinib and celecoxib groups (P>0.05). Western blotting showed decreased expression of P-EGFR and COX-2 with both erlotinib and celecoxib treatments, but most pronouncedly in the combined group (P<0.05). Simultaneous blockage of the EGFR and COX-2 signal pathways exerted stronger growth effects in our human xenotransplanted lung cancer model than inhibition of either pathway alone. The anti-tumor effects were accompanied by synergetic inhibition of tumor cell apoptosis, activation of p-EGFR and expression of COX-2.