Evaluation of a novel saliva-based epidermal growth factor receptor mutation detection for lung cancer: A pilot study.

BackgroundThis article describes a pilot study evaluating a novel liquid biopsy system for non-small cell lung cancer (NSCLC) patients. The electric field-induced release and measurement (EFIRM) method utilizes an electrochemical biosensor for detecting oncogenic mutations in biofluids.MethodsSaliva and plasma of 17 patients were collected from three cancer centers prior to and after surgical resection. The EFIRM method was then applied to the collected samples to assay for exon 19 deletion and p.L858 mutations. EFIRM results were compared with cobas results of exon 19 deletion and p.L858 mutation detection in cancer tissues.ResultsThe EFIRM method was found to detect exon 19 deletion with an area under the curve (AUC) of 1.0 in both saliva and plasma samples in lung cancer patients. For L858R mutation detection, the AUC of saliva was 1.0, while the AUC of plasma was 0.98. Strong correlations were also found between presurgery and post-surgery samples for both saliva (0.86 for exon 19 and 0.98 for L858R) and plasma (0.73 for exon 19 and 0.94 for L858R).ConclusionOur study demonstrates the feasibility of utilizing EFIRM to rapidly, non-invasively, and conveniently detect epidermal growth factor receptor mutations in the saliva of patients with NSCLC, with results corresponding perfectly with the results of cobas tissue genotyping

Long-term Survival of Personalized Surgical Treatment of Locally Advanced Non-small Cell Lung Cancer Based on Molecular Staging

Background and objective Approximately 35%-40% of patients with newly diagnosed non-small cell Lung cancer have locally advanced disease. The average survival time of these patients only have 6-8 months with chemotherapy. The aim of this study is to explore and summarize the probability of detection of micrometastasis in peripheral blood for molecular staging, and for selection of indication of surgical treatment, and beneficiary of neoadjuvant chemotherapy and postoperative adjuvant therapy in locally advanced lung cancer; to summarize the long-time survival result of personalized surgical treatment of 516 patients with locally advanced non-small cell lung cancer based on molecular staging methods. Methods CK19 mRNA expression of peripheral blood samples was detected in 516 lung cancer patients by RT-PCR before operation for molecular diagnosis of micrometastasis, personalized molecular staging, and for selection of indication of surgical treatment and the beneficiary of neoadjuvant chemotherapy and postoperative adjuvant therapy in patients with locally advanced nonsmall cell lung cancer invaded heart, great vessels or both. The long-term survival result of personalized surgical treatment was retrospectively analyzed in 516 patients with locally advanced non-small cell lung cancer based on molecular staging methods. Results There were 322 patients with squamous cell carcinoma and 194 cases with adenocarcinoma in the series of 516 patients with locally advanced lung cancer involved heart, great vessels or both. There were 112 patients with IIIA disease and 404 cases with IIIB disease according to P-TNM staging. There were 97 patients with M-IIIA disease, 278 cases with M-IIIB disease and 141 cases with III disease according to our personalized molecular staging. Of the 516 patients, bronchoplastic procedures and pulmonary artery reconstruction was carried out in 256 cases; lobectomy combined with resection and reconstruction of partial left atrium was performed in 41 cases; Double sleeve lobectomy combined with resection and reconstruction of super vena cava was carried out in 90 cases; Lobectomy combined with resection and reconstruction of diaphragm was performed in 3 cases; Double sleeve lobectomy combined with resection and reconstruction of partial left atrium was performed in 30 cases; Bronchoplastic procedures and pulmonary artery reconstruction combined with reconstruction of aorta sheath was carried out in 10 cases; Right pneumonectomy combined with resection and reconstruction partial left atrium, total right diaphragm with Dacron, and post cava and right liver vein was performed in one case; Lobectomy combined with resection and reconstruction of carina was carried out in 10 cases; Bronchoplastic procedures and pulmonary artery reconstruction combined with resection and reconstruction of carina and superior vane cava, or combined with superior vena cava and left atrium, or with carina and left atrium was performed in 55 cases in this series. Five patients died of operative complications and the operative mortality was 0.97%. CK19 mRNA expression was found in 141 patients. The positive rate of CK19 mRNA expression was 27.3% in peripheral blood samples in the 516 cases. The positive rates of micrometastasis in peripheral blood was significantly related to histological classification, P-TNM staging and N staging of the cancer (P < 0.05), but not to age, sex, smoking status of the patients, and size of primary tumor, and locations of the tumor (P > 0.05). The median survival time was 43.74 months. The 1, 3, 5 and 10 year survival rates of the 516 cases was 89.1%, 39.3%, 19.8% and 10.4%, respectively. The postoperative survival rate was remarkably correlated with micrometastasis in peripheral blood, histological classification of the tumor, size of primary cancer and lymph mode involvement (P < 0.05). The results of multivariable Cox model analysis showed that "personalizedmolecular P-TNM staging", micrometastasis in peripheral blood, pathological types of the tumor and mediastinal lymph node metastasis of the cancer were the most significant factors for predicting prognosis in the patients with locally advanced nonsmall lung cancer. Conclusion (1) Micrometastasis was existed in peripheral blood of patients with lung cancer, which can not be detected with conventional methods. (2) Detecting of CK19 mRNA expression in peripheral blood in lung cancer patients can be used for diagnosis of micrometastasis of lung cancer and “molecular staging” and “molecular P-TNM staging” for lung cancer patients. It will be helpful for selection of surgical treatment indication, the beneficiary of neoadjuvant chemotherapy and postopertive adjuvant therapy in the patients with locally advanced non-small cell lung cancer. (3) Personalized surgical treatment can significantly improve prognosis and increase curative rate and long-term survival rate of locally advanced nonsmall cell lung cancer based on personalized molecular staging

覚せい剤精神病脆弱性に於けるドーパミンD2、セロトニンS2受容体のポジトロンCTによる測定

研究科: 千葉大学大学院医学研究科千大院医博乙第1281

Single Endemic Genotype of Measles Virus Continuously Circulating in China for at Least 16 Years

The incidence of measles in China from 1991 to 2008 was reviewed, and the nucleotide sequences from 1507 measles viruses (MeV) isolated during 1993 to 2008 were phylogenetically analyzed. The results showed that measles epidemics peaked approximately every 3 to 5 years with the range of measles cases detected between 56,850 and 140,048 per year. The Chinese MeV strains represented three genotypes; 1501 H1, 1 H2 and 5 A. Genotype H1 was the predominant genotype throughout China continuously circulating for at least 16 years. Genotype H1 sequences could be divided into two distinct clusters, H1a and H1b. A 4.2% average nucleotide divergence was found between the H1a and H1b clusters, and the nucleotide sequence and predicted amino acid homologies of H1a viruses were 92.3%–100% and 84.7%–100%, H1b were 97.1%–100% and 95.3%–100%, respectively. Viruses from both clusters were distributed throughout China with no apparent geographic restriction and multiple co-circulating lineages were present in many provinces. Cluster H1a and H1b viruses were co-circulating during 1993 to 2005, while no H1b viruses were detected after 2005 and the transmission of that cluster has presumably been interrupted. Analysis of the nucleotide and predicted amino acid changes in the N proteins of H1a and H1b viruses showed no evidence of selective pressure. This study investigated the genotype and cluster distribution of MeV in China over a 16-year period to establish a genetic baseline before MeV elimination in Western Pacific Region (WPR). Continuous and extensive MeV surveillance and the ability to quickly identify imported cases of measles will become more critical as measles elimination goals are achieved in China in the near future. This is the first report that a single endemic genotype of measles virus has been found to be continuously circulating in one country for at least 16 years

Combined Double Sleeve Lobectomy and Superior Vena Cava Resection for Non-small Cell Lung Cancer with Persistent Left Superior Vena Cava

A 65-year-old man with right central type of lung squamous carcinoma was admitted to our department. Bronchoscopy displayed complete obstruction of right upper lobe bronchus and infiltration of the bronchus intermedius with tumor. Chest contrast computed tomography revealed the tumor invaded right pulmonary artery, superior vena cava, and the persistant left superior vena cava flowed into the coronary sinus. The tumor was successfully removed by means of bronchial and pulmonary artery sleeve resection of the right upper and middle lobes combined with resection and reconstruction of superior vena cava (SVC) utilizing ringed polytetrafluoroethylene graft. To the best of our knowledge, this was the first report of complete resection of locally advanced lung cancer involving superior vena cava, right pulmonary artery trunk and main bronchus with persistant left superior vena cava

Seismic forward and inverse simulation in a tight reservoir model of loess plateau region

To find out the seismic wave field propagation principles in loess plateau near surface of the Ordos Basin and the seismic response characteristics of tight oil reservoirs, this study established a geological-geophysical model under the real conditions of ground surface of loess plateau, and launched full elastic seismic wave equation forward modeling and pre-stack elastic seismic inversion study. Comparison of modeling and real seismic data shows that, the loose and wavy loess plateau surface is the main reason for causing the problems of seismic static correction and interference wave. Tomographic static correction method with the constraint of traces near shot point can effectively solve the problem of seismic static correction in the loess plateau and enhance seismic imaging accuracy, S-wave impedance obtained from pre-stack seismic inversion can identify sandstone effectively, and Poisson's ratio can identify oil-bearing reservoirs. The seismic forward and inverse simulation and rock physical analysis provide a solid theoretical and experimental basis for the seismic prediction of tight oil reservoir, and have worked well in the oil exploration and development in the loess plateau of the Ordos Basin. Key words: loess plateau surface, tight reservoir, wave equation forward simulation, tomographic inversion static correction, pre-stack elastic inversio

Immunoglobulin Light Chain Use and Characteristics in the Chlamydophila Psittaci negative MALT Lymphomas of the Ocular Adnexa

Abstract Abstract 1597 Ocular adnexal mucosa associated lymphoid tissue lymphomas (OAMALTL) are the most common lymphomas of the eye. The potential roles for specific antigens in these lymphomas are still controversial. Previously we examined the usage and mutations of the IGVH in Chlamydophila (C) psittaci-negative OAMALTL, demonstrating biased use of the IGHV4 family and IGHV4–34 gene and evidence for antigen selection (PLoS One. 2011;6(12):e29114). However, there have been no previous reports characterizing the Ig light chain (IGKV/IGLV) usage and properties in OAMALTL. Herein, we examined IGKV/IGLV gene usage and mutations in 34 C. psittaci-negative OAMALTL originating from the orbit (15), conjunctivae (14) and lacrimal gland (5). Clonal potentially functional IGKV/IGLV gene sequences were identified in 30 tumors (18 kappa and 12 lambda). Among the 7 kappa light chain families, IGKV1 (n=5, 28%), IGKV2 (n=1, 6%), IGKV3 (n=8, 44%) and IGKV4 (n=4, 22%) were observed. Among the 8 lambda chain families, IGLV2 (n=5, 42%), IGLV3 (n=1, 8%), IGLV6 (n=1, 8%), IGLV7 (n=3, 25%) and IGLV8 (n=2, 17%) were detected. In comparison with the IGKV and IGLV repertoire employed in normal peripheral blood B-lymphocytes, an overrepresentation of the IGKV4 family (P<0.01) was observed in the analyzed cohort of tumors. No statistically significant difference in the use of other kappa and lambda light chain families was observed. In the most commonly used IGKV3 family, only 2 (IGKV3–20*01 (n=6) and IGKV3–15*01 (n=2)) of the 7 family members were used by the OAMALTL. The IGKV3–20*01 allele was used at a greater frequency than in normal peripheral blood B-lymphocytes (p=0.02). The observed frequency of the allele in OAMALTL (20%) is higher than the previously reported frequency in other non-MALT lymphoma types. Of the six cases which utilized the IGKV3–20*01 allele, corresponding heavy chains were available for five cases. Three of the IGKV3–20*01 alleles paired with the IGHV4–34 allele, one paired with the IGHV3–23 allele and one paired with the IGHV1–69 allele. Interestingly, each of these three alleles has been previously implicated in autoreactivity. The IGKV4-1*01 allele was observed to pair with IGHV3–74 (n=2), IGHV1–24 (n=1) and IGHV2–5 (n=1), respectively. Of the 30 clonal tumor sequences, 27 displayed mutations from germline. The average percent homology to germline was 96.35%. Most mutated cases (14 out of 27, 52%) exhibited less than a 2% difference from their original germline sequence. Four of the six tumors utilizing the IGKV3–20*01 allele exhibited more than 2% mutation away from germline, with the remaining two sequences unmutated. Three of the four tumors employing IGKV4-1*01 exhibited greater than 2% mutation away from germline. No acquired N-glycosylation sites were observed in the OAMALTL. To analyze for potential selective pressure exerted by antigens, we utilized the BASELINe algorithm, which assesses for both presence and strength of antigen selection. 23 sequences exhibited evidence of selection in the CDR regions, while 26 sequences exhibited evidence of selection in the FR regions. Of the 26 sequences for which paired light and heavy chains were available, 9 heavy chains exhibited strong evidence of selection (8 in FR, 1 in CDR) and 5 light chain exhibited strong evidence of selection (all in FR). Two patients' tumors exhibited strong evidence of selection in both heavy and light chains. A recurrent mutation was observed in the FR2 region of sequences derived from the IGKV3–20*01, with three of six cases showing an acquisition of histidine in place of glutamine at position 39. An analysis of the tumor derived CDR3 regions did not reveal stereotyped sequence determinants and showed no homology to other known bacterial binding antibodies. Our findings demonstrate that C. psittaci negative OAMALTL exhibit biased usage of IGKV families and genes with evidence of antigen selection. Combined with our previous findings, we demonstrate the frequent use and common pairing of IGVH and IGKV genes, which are usually used by autoantibodies, suggesting that the C. psittaci negative OAMALTL encoded immunoglobulins may be directed to autoantigens. Indeed, generated recombinant immunoglobulins derived from multiple OAMALTLs react to self and not to bacterial antigens. These findings and the identity of the autoantigens will be presented at the meeting. Disclosures: No relevant conflicts of interest to declare

The Study on the Phosphorylation Activity of Mutant nm23-H1

Background and objective The Phosphorylation is the key activity of nm23-H1. The aim of this study is to explore the effect of different amino acid mutation on the phosphorylation status of nm23-H1. Methods The wild type nm23-H1 was as the control of this study. Autoradiography was used for detecting the serine and histidineautophosphorylation of wild type (WT) and mutant nm23-H1 (P96S, H118F, S120G and S44A); RP-HPLC was used for detecting the NDPK activity of above proteins. Results The autophosphorylation activities of serine and histidine from high to low were P96S, WT, S44A, S120G and H118F, respectively, while the NDPK activities from high to low were WT, S120G, P96S, S44A, H118F. A highly positive correlation was found between serine and histidine autophosphorylation activity of above proteins (r =0.985, P 0.05, and r =0.482, P >0.05, respectively). Conclusion Site mutation of nm23-H1 can affect the phosphorylation activity. H118 site was the key amino acid of kinase activity, P96 site maybe related to phosphotransferring, S120 was the site of histidine autophosphorylation and serine autophosphorylation, while the S44 site may be another amino acid which possessed NDPK activity