Predelivery HbA1c levels and their relationship with adverse perinatal outcomes in women with normal 75-g OGTT.

PURPOSE: To analyze the elevation of HbA1c before full-term delivery in single pregnancy with normal 75 g- Oral glucose tolerance test (OGTT) screening and its association with adverse perinatal outcomes. METHODS: From January to December 2022, an observational prospective study was conducted in a Single centre in China. 365 single pregnant women with normal OGTT were included in the study. HbA1c was measured during OGTT and before full-term delivery, and perinatal outcomes were collected. Participants' pre-delivery HBA1c values were analyzed and perinatal outcomes were compared. Logistic regression analysis was used for independent risk factors associated with elevated pre-delivery HbA1c ≥ 6.0%. The predictive value and truncation value were analyzed by ROC curve. RESULTS: 15.89% (58/365) of the Participants had a pre-delivery HBA1C value ≥ 6.0%. The incidence of neonatal asphyxia (13.79%, vs. 3.45%, vs. 2.26%, P = 0.007) and amniotic fluid fecal staining (29.31%, vs. 12.64%, vs. 12.03%, P = 0.004) were significantly increased in this group. The independent risk factor associated with pre-delivery HBA1c ≥ 6.0% was the fasting blood glucose(FBG) value of OGTT (OR = 51.308, 95% CI 12.93-203.67, P < 0.01) and the HBA1c value measured during OGTT (OR = 3.608, 95% CI 1.432-9.151, P = 0.007). When FBG was < 4.18 mmol/L and HBA1c was < 5.51%, The accuracy of predicting the pre-delivery HBA1c < 6.0% was 98.2%. CONCLUSIONS: 15.89% of the single pregnancy with normal OGTT had HbA1c ≥ 6.0% before full-term delivery, and they had an increased incidence of neonatal asphyxia and amniotic fluid fecal staining. When the FBG ≥ 4.18 mmol/l or the HBA1c ≥ 5.51% during the OGTT screening, repeated OGTT were recommended in late pregnancy.

The efficacy of Raman spectroscopy in lung cancer diagnosis: the first diagnostic meta-analysis.

In recent years, many researches have explored the diagnostic value of Raman spectroscopy in multiple types of tumors. However, as an emerging clinical examination method, the diagnostic performance of Raman spectroscopy in lung cancer remains unclear. Relevant diagnostic studies published before 1 June 2020 were retrieved from the Cochrane Library, PubMed, EMBASE, China National Knowledge Internet (CNKI), and WanFang databases. After the literature was screened, two authors extracted the data from eligible studies according to the inclusion and exclusion criteria. Obtained data were pooled and analyzed using Stata 16.0, Meta-DiSc 1.4, and RevMan 5.3 software. Fourteen diagnostic studies were eligible for the pooled analysis which includes 779 patients. Total pooled sensitivity and specificity of Raman spectroscopy in diagnosing lung cancer were 0.92 (95% CI 0.87-0.95) and 0.94 (95% CI 0.88-0.97), respectively. The positive likelihood ratio was 15.2 (95% CI 7.5-30.9), the negative likelihood ratio was 0.09 (95% CI 0.05-0.14), and the area under the curve was 0.97 (95 % CI 0.95-0.98). Subgroup analysis suggested that the sensitivity and specificity of RS when analyzing human tissue, serum, and saliva samples were 0.95 (95% CI 0.88-0.98), 0.97 (95% CI 0.89-0.99), 0.88 (95% CI 0.80-0.93), 0.87 (95% CI 0.78-0.92), 0.91 (95% CI 0.80-0.96), and 0.95 (95% CI 0.73-0.99), respectively. No publication bias or threshold effects were detected in this meta-analysis. This initial meta-analysis indicated that Raman spectroscopy is a highly specific and sensitive diagnostic technology for detecting lung cancer. Further investigations are also needed to focus on real-time detection using Raman spectroscopy under bronchoscopy in vivo. Moreover, large-scale diagnostic studies should be conducted to confirm this conclusion.

The diagnostic value of Hsp90α in monitoring treatment responses in lung cancer.

BACKGROUND: Heat shock protein 90α (Hsp90α) is considered a tumor biomarker in many human malignancies. This study investigated the diagnostic value of Hsp90α combined with other traditional lung cancer biomarkers (CEA, CYFRA21-1, and NSE) and its role in monitoring the treatment response of lung cancer patients. METHODS: A total of 205 patients with lung cancer and 186 patients with lung benign disease who were admitted to our hospital were enrolled from 2018 to 2020. The 205 patients included 76 cases of squamous, 92 cases of adenocarcinoma, and 37 cases of small cell lung cancer. There were 49 patients with TNM I+II and 156 patients with TNM III+IV. A total of 10 mL baseline peripheral venous blood samples and subsequent peripheral venous blood samples (7 days after two cycles of chemotherapy) were collected, and the levels of Hsp90α, carcinoembryonic antigen (CEA), Cytokeratin 19 fragments (CYFRA21-1), and neuron-specific enolase (NSE) were detected by ELISA kit. RESULTS: Hsp90α was obviously higher in serum from patients with lung cancer than in patients with benign lung disease (p < 0.0001). Moreover, Hsp90α levels were higher in patients with advanced-stage (stage III-IV) lung cancer compared to those with early-stage (stage I-II). Hsp90α level was significantly decreased following treatment with chemotherapy in the progress partial response group (p = 0.017), whereas the level of Hsp90α was significantly higher after chemotherapy treatment in the progressive disease group (p < 0.0001). In addition, compared with CYFRA21-1, CEA, or NSE alone, the AUC of Hsp90α combined with CYFRA21-1, CEA, or NSE were significantly higher in the diagnosis of adenocarcinoma or small-cell lung cancer. DISCUSSION: Hsp90α combined with CYFRA21-1, CEA, and NSE can be used as diagnostic indicators of lung cancer. The Hsp90α level can be used to monitor treatment response.

Sarcoidosis Stage II: a Case Report and Review of the Literature.

BACKGROUND: Sarcoidosis is a systemic granulomatous disease of unknown origin characterized by non-caseous necrotizing epithelial cell granuloma that affects the lung and lymphatic system. Sarcoidosis mainly occurs in young and middle-aged people, usually manifested as bilateral hilar lymph node enlargement, lung infiltration, and eye and skin lesions. Sarcoidosis has a high natural remission rate, but patients with progressive imaging or pulmonary function accompanied by significant clinical symptoms or extrapulmonary lesions need to be treated. METHODS: The sarcoidosis patient had received a 3-month methylprednisolone treatment which significantly improved clinical manifestations including cough and sputum, and extrapulmonary presentation, such as skin nodules and enlargement of parotid glands. RESULTS: A 52-year-old female reporting repeated cough and sputum, with scattered skin rashes and nodules on the extremities, accompanied by nasal congestion, enlargement of abdominal and retroperitoneal lymph nodes and parotid glands was studied. Computed tomography (CT) showed miliary nodules diffusely distributed in both lungs, multiple enlarged lymph nodes in mediastinum, bilateral enlarged hilar lymph nodes, and right pleural effusion. Bronchoscopy with lung biopsy showed granuloma formation, special staining including acid resistance was negative, but signet ring cell carcinoma and tuberculosis cannot be excluded. Biopsy of a skin nodule also showed granulomatosis. PET-CT reported all considered as inflammatory lesions, with a high possibility of tuberculosis. Based on all the information, we confirmed the diagnosis of sarcoidosis stage II. She was then successfully treated with a steroid monotherapy, which resulted in a satisfactory clinical outcome without serious complications. CONCLUSIONS: Clinical manifestations of this patient are unspecific. Based on the pathological finding, clinical and radiological manifestation, and evidence of no alternative diseases, sarcoidosis stage II is diagnosed. Treatment with a steroid was of benefit in this sarcoidosis patient.

Hypoxia promotes mitochondrial glutamine metabolism through HIF1α-GDH pathway in human lung cancer cells.

Drug-resistance is common in human lung cancer therapy. Hypoxia remarkably contributes to drug-resistance in lung cancer but the underlying mechanism remains elusive. Here we demonstrate that hypoxia-induced glutamine metabolism is involved in drug resistance in lung cancer cells. Hypoxia increases glutamine up-take, glutamate to α-ketoglutarate flux and the generation of ATP in lung cancer cells by up-regulating the expression of glutamate dehydrogenase (GDH). Hypoxia-induced expression of GDH relies on the up-regulation of HIF1α but not HIF2α. HIF1α binds the promoter of GDH and promotes the transcription of GDH gene in lung cancer cells. Finally, we show that GDH represses cisplatin-induced cell apoptosis and repression of colony formation, indicating that GDH contributes to drug-resistance in lung cancer cells. In conclusion, HIF1α-GDH pathway regulates glutamine metabolism and ATP production upon hypoxia stress and contributes to drug-resistance in human lung cancer cells.

Gemcitabine-Induced Autophagy Protects Human Lung Cancer Cells from Apoptotic Death.

PURPOSE: Gemcitabine has been used as a therapeutic drug combined with cisplatin for the treatment of lung cancer patients. However, the prognosis is poor due to acquired resistance. Accumulating studies have revealed that autophagy may contribute to the drug resistance. Therefore, the present study is aimed to clarify the mechanisms underlying gemcitabine-acquired resistance. METHODS: SPC-A1 and A549 cells were incubated with gemcitabine followed by assessment of cell viability with MTT assays. GFP-LC3 transient transfection, MDC staining, and transmission electron microscopy were used to detect the change of autophagy at morphological level. Flow cytometry was used to monitor the effect of 3-MA on gemcitabine-induced apoptosis. Western blot analysis was used to detect the expression of p62, LC3, Beclin-1, ATG5, activated caspase 3, Bax, BNIP3, BNIP3L, and Bcl-2. RESULTS: Our study showed that gemcitabine significantly induced both autophagy and apoptosis in human lung cancer cells SPC-A1 and A549. Of interest was that when autophagy was inhibited by 3-MA, the gemcitabine-induced apoptosis was effectively enhanced, suggesting that gemcitabine can activate autophagy to impair the chemosensitivity of lung cancer cells. Furthermore, the inhibition of autophagy by 3-MA further increased the expression of activated caspase 3, Bax, BNIP3, and BNIP3L, all are critical apoptotic mediators. Contrarily, 3-MA treatment further decreased the expression of Bcl-2, which is an important anti-apoptotic protein. CONCLUSION: Our study indicated that autophagy protected human lung cancer cells from gemcitabine-induced apoptosis, and the combined use of gemcitabine and an autophagic inhibitor in lung cancer patients may be an effective therapeutic strategy.

Pedicle drilling force control of a robotic surgical system via spine-soft tissue coupling model and parameters optimization.

Bone drilling is a crucial operation in spinal fusion surgery that requires precise control of the applied force to ensure surgical safety. This manuscript aims to enhance the force servo performance of the orthopedic robot during automatic bone drilling operations. Firstly, an analytical model is introduced to describe the spinal mobility of the spine-soft tissue coupling structure. Then, the model is calibrated using force data obtained from stress relaxation tests. Next, optimal force controller parameters are determined through drilling force control simulations based on the identified model. The dynamic performance and robustness of the closed-loop control system are analyzed to ensure safe drilling procedures. Finally, bone drilling experiments are conducted in a force control mode to verify the effectiveness of the proposed method. The step drilling force response's steady-state error is less than 0.15 N, the relative control error is less than 3 %, and there is no noticeable force overshoot. The amplitude of the sinusoidal force response decays to -3 dB when the target force frequency is up to 3.49 rad/s, indicating a wide control bandwidth. These results demonstrate that the proposed method can rapidly and safely provide an adequate force servo to carry out automatic bone drilling.

Inkjet printer prediction under complicated printing conditions based on microscopic image features.

A novel technique is introduced to predict the printer model used to produce a given document. Samples containing only a few letters printed under varying conditions (i.e., different printing modes, letter types, fonts) were collected to establish a dataset of 41 inkjet printer models from common manufacturers, such as HP, Canon, and Epson. Morphological features were analyzed by extraction of image features using several algorithms in a series of microscopic images and a Wilcoxon test was used to measure the significance of variations between printed samples. Significant differences between various printing conditions might post potential challenge to questioned document examination. Discriminant analysis and the k-nearest neighbor (KNN) algorithm were also employed for source printer prediction under varying printing condition on 30% images with the rest images as training dataset. The results of a validation experiment demonstrated that while quadratic discriminant analysis (QDA) achieved an accuracy of 96.3%, a combination of KNN and QDA reached 98.6%. As such, this technique could aid in the forensic examination of printed documents.

Prediction of laser printers and cartridges based on three-dimensional profiles via discrimination analysis.

Printer source prediction is an important task when examining questioned documents. While some research has provided methods to predict the source printer of documents, with the advent of compatible consumables, printer prediction could become more complex and difficult. Predicting the source printer after replacing cartridges and identifying the source of printer cartridges are unresolved issues that are rarely addressed in current research. Herein, we introduce a novel technique to predict the manufacturer, model, and cartridges of laser printers (i.e., compatible, and original cartridges) used to produce a given document. Document samples produced using eight laser printers and 247 cartridges were collected to establish a dataset. Common manufacturers included HP, Canon, Lenovo, and Epson. After obtaining white-light images and three-dimensional profile images of printed characters, a morphological analysis was conducted by questioned document examiners (QDEs) using microscopy. Microscopic image features across a series of images were also extracted and analyzed using algorithms. Then, six high-dimensional reduction algorithms were used to obtain between- and within-printer variations as well as between- and within-cartridge variations. Finally, we conducted principal component analysis (PCA) and discriminant analysis. For 40 % of the samples, mixed discrimination analysis (MDA) and fixed discrimination analysis (FDA) were employed to predict the manufacturer, model and cartridge of laser printers used to produce the questioned printed document; the remaining 60 % samples comprised the training dataset. In the prediction of manufacturer, model and cartridge, our method achieved mean accuracies of 95.5 %, 97.5 %, and 90.2 %, respectively. Hence, this technique could reasonably aid in predicting the manufacturer, model, and cartridge of a laser printer, even if different cartridges are loaded into printers.

Comparison of the genome sequences of "Candidatus Portiera aleyrodidarum" primary endosymbionts of the whitefly Bemisia tabaci B and Q biotypes.

"Candidatus Portiera aleyrodidarum" is the primary endosymbiont of whiteflies. We report two complete genome sequences of this bacterium from the worldwide invasive B and Q biotypes of the whitefly Bemisia tabaci. Differences in the two genome sequences may add insights into the complex differences in the biology of both biotypes.

Genome sequences of the primary endosymbiont "Candidatus Portiera aleyrodidarum" in the whitefly Bemisia tabaci B and Q biotypes.

"Candidatus Portiera aleyrodidarum" is the obligate primary endosymbiotic bacterium of whiteflies, including the sweet potato whitefly Bemisia tabaci, and provides essential nutrients to its host. Here we report two complete genome sequences of this bacterium from the B and Q biotypes of B. tabaci.

Enrichment of mRNA-like noncoding RNAs in the divergence of Drosophila males.

With the advent of transcriptome data, it has become clear that mRNA-like noncoding RNAs (mlncRNAs) are widespread in eukaryotes. Although their functions are poorly understood, these transcripts may play an important role in development and could thus be involved in determining developmental complexity and phenotypic diversification. However, few studies have assessed their potential roles in the divergence of closely related species. Here, we identify and study patterns of sequence and expression divergence in ten novel candidate mlncRNAs from Drosophila pseudoobscura and its close relative D. persimilis. The candidate mlncRNAs were identified by randomly sequencing a group of 734 cDNA clones from a microarray that showed either no difference in expression (187 clones) or differential expression (547 clones) in comparisons between D. pseudoobscura and D. persimilis and between these two species and their F(1) hybrids. Candidate mlncRNAs are overrepresented among differentially expressed transcripts between males of D. pseudoobscura and D. persimilis, and although they have high sequence conservation between these two species, seven of them have no putative homologs in any of the other ten Drosophila species whose genomes have been sequenced. Expression of eight of the ten candidate mlncRNAs was detected either in whole bodies (adults) or testes using a custom-designed oligonucleotide microarray. Three of the ten candidate mlncRNAs are highly expressed (in the top 4% of the male transcriptome), differentially expressed between species, and show extreme levels of sex-bias, with one transcript having the highest level of male bias in the whole transcriptome. Proteomic data from testes show no traces of any predicted peptides from the candidate mlncRNAs. Our results suggest that these mlncRNAs may be important in male-specific processes related to sexual dimorphism and species divergence in this species group.

Decreased expression of Beclin-1 and LC3 in human lung cancer.

Autophagy, a conversed response to stress, has recently been studied in human cancers. Two important autophagic genes-Beclin-1 and LC3 are reported in several human cancers. However, the expressions of Beclin-1 and LC3 in lung cancer have not yet been investigated. In the present study, we investigated the expression of Beclin-1 and LC3, and the relationship between the expression profile and the clinical or pathological changes in human lung cancer. 40 primary lung cancer patients are involved in present study. mRNA expressions of Beclin-1 and LC3-II were detected by Real Time PCR and the protein levels were assessed by immunohistochemistry and western blot. Relative lower expressions of Beclin-1 and LC3-II mRNA were found in the lung cancer tissues compared to counterpart normal tissues. Consistently, the lower amount of Beclin-1 and LC3-II protein was found in lung cancer tissues. However, the expressions of Beclin-1 and LC3-II in lung cancer tissues were not affected by patients' age, gender, smoking, histological type, lymph node metastasis and tumor-node-metastasis (TNM) stage. Both mRNA and protein levels of Beclin-1 and LC3-II were significantly decreased in lung cancer tissues which suggested that autophagy may be involved in the pathogenesis of lung cancer.

Occupational fibrotic hypersensitivity pneumonia in a halogen dishes manufacturer: A case report.

BACKGROUND: Fibrotic hypersensitivity pneumonitis (FHP) is an allergic and diffuse pneumonia caused by repeated inhalation of antigenic substances, and sometimes developed in people working in specific environments. While novel antigens and exposures continued to be described, physicians should maintain a high suspicion of potential exposures. A detailed assessment of the patient's occupational exposures as well as living environment is necessary and complete allergen avoidance is the first and most important step in the management of FHP once the allergens are determined. CASE SUMMARY: A 35-year-old female was admitted to the hospital with a cough and breathing difficulties for more than one year. She was a nonsmoker and a manufacturer of halogen dishes, which are characteristic Chinese foods, for 15 years without any protection. High resolution computed tomography of the chest demonstrated an interstitial pneumonia pattern. Pulmonary function examination showed restricted ventilation dysfunction and a significant reduction in dispersion ability. Cell differentiation in bronchoalveolar lavage fluid demonstrated lymphocytosis (70.4%) with an increased lymphocyte CD4/CD8 ratio (0.94). Transbronchial lung biopsy combined with lung puncture pathology showed diffuse uniform alveolar interval thickening, chronic inflammatory cell infiltration, a proliferation of tissue in the bronchial wall fiber and alveolar epithelial follicle degeneration, resulting in fibrosis. CONCLUSION: Exposure to spices used for the production of halogen dishes may cause FHP.

Giant tortoise genomes provide insights into longevity and age-related disease.

Giant tortoises are among the longest-lived vertebrate animals and, as such, provide an excellent model to study traits like longevity and age-related diseases. However, genomic and molecular evolutionary information on giant tortoises is scarce. Here, we describe a global analysis of the genomes of Lonesome George-the iconic last member of Chelonoidis abingdonii-and the Aldabra giant tortoise (Aldabrachelys gigantea). Comparison of these genomes with those of related species, using both unsupervised and supervised analyses, led us to detect lineage-specific variants affecting DNA repair genes, inflammatory mediators and genes related to cancer development. Our study also hints at specific evolutionary strategies linked to increased lifespan, and expands our understanding of the genomic determinants of ageing. These new genome sequences also provide important resources to help the efforts for restoration of giant tortoise populations.

Poly-L-Arginine Acts Synergistically with LPS to Promote the Release of IL-6 and IL-8 via p38/ERK Signaling Pathways in NCI-H292 Cells.

Major basic protein (MBP) derived from activated eosinophil can exacerbate atopic asthma induced by lipopolysaccharide (LPS). The pharmacological function of MBP can be mimicked by poly-L-arginine (PLA), however, the potential signaling mechanisms of LPS-PLA-induced release of the inflammatory cytokines interleukin (IL)-6 and IL-8 remain unclear. In the present study, airway epithelia NCI-H292 cell lines were treated with LPS and/or PLA. We found that the expression levels of IL-6 and IL-8 induced by LPS-PLA were increased significantly compared with that in untreated cells. Meanwhile, the phosphorylation of p38 MAPK and ERK1/2 was also up-regulated dramatically by LPS-PLA, but this increase could be blocked by specific inhibitor. Importantly, blocking the phosphorylation of p38 MAPK and ERK1/2 reduced the expression levels of IL-6 and IL-8 as well. Collectively, LPS-PLA-induced release of IL-6 and IL-8 from NCI-H292 cells may be due to the synergistic activation of p38 MAPK and ERK1/2 signaling transduction pathways.

Gene silencing of USP1 by lentivirus effectively inhibits proliferation and invasion of human osteosarcoma cells.

Osteosarcoma is the most frequent malignant bone tumor, affecting the extremities of adolescents and young adults. Ubiquitin-specific protease 1 (USP1) plays a critical role in many cellular processes including proteasome degradation, chromatin remodeling and cell cycle regulation. In the present study, we discovered that USP1 was overexpressed in 26 out of 30 osteosarcoma tissues compared to cartilage tumor tissues and normal bone tissues. We then constructed a lentiviral vector mediating RNA interference (RNAi) targeting USP1 and demonstrated that it significantly suppressed the mRNA and protein expression of the USP1 gene in U2OS cells. Knockdown of USP1 inhibited the growth and colony-forming, as well as significantly reduced the invasiveness of U2OS cells. Western blot analysis indicated that suppression of USP1 downregulated the expression of many proteins including SIK2, MMP-2, GSK-3ß, Bcl-2, Stat3, cyclin E1, Notch1, Wnt-1 and cyclin A1. Most of these proteins are associated with tumor genesis and development. RNAi of SIK2 significantly decreased SIK2 protein expression and inhibited the ability of forming colonies, as well as induced apoptosis and reduced the invasiveness of U2OS cells. Collectively, our results suggest that silencing USP1 inhibits cell proliferation and invasion in U2OS cells. Therefore, USP1 may provide a novel therapeutic target for the treatment of osteosarcoma.

Hypoxia-induced autophagy mediates cisplatin resistance in lung cancer cells.

Hypoxia which commonly exists in solid tumors, leads to cancer cells chemoresistance via provoking adaptive responses including autophagy. Therefore, we sought to evaluate the role of autophagy and hypoxia as well as the underlying mechanism in the cisplatin resistance of lung cancer cells. Our study demonstrated that hypoxia significantly protected A549 and SPC-A1 cells from cisplatin-induced cell death in a Hif-1α- and Hif-2α-dependent manner. Moreover, compared with normoxia, cisplatin-induced apoptosis under hypoxia was markedly reduced. However, when autophagy was inhibited by 3-MA or siRNA targeted ATG5, this reduction was effectively attenuated, which means autophagy mediates cisplatin resisitance under hypoxia. In parallel, we showed that hypoxia robustly augmented cisplatin-induced autophagy activation, accompanying by suppressing cisplatin-induced BNIP3 death pathways, which was due to the more efficient autophagic process under hypoxia. Consequently, we proposed that autophagy was a protective mechanism after cisplatin incubation under both normoxia and hypoxia. However, under normoxia, autophagy activation 'was unable to counteract the stress induced by cisplatin, therefore resulting in cell death, whereas under hypoxia, autophagy induction was augmented that solved the cisplatin-induced stress, allowing the cells to survival. In conclusion, augmented induction of autophagy by hypoxia decreased lung cancer cells susceptibility to cisplatin-induced apoptosis.

Amplicon Indel Hunter Is a Novel Bioinformatics Tool to Detect Large Somatic Insertion/Deletion Mutations in Amplicon-Based Next-Generation Sequencing Data.

Amplicon-based targeted next-generation sequencing assays are used widely to test for clinically relevant somatic mutations in cancer. However, accurate detection of large insertions and deletions (indels) via these assays remains challenging. Sequencing reads that cover these anomalies are, by definition, different from the reference sequence, and lead to variable performance of alignment algorithms. Reads with large indels may be aligned incorrectly, causing incorrect calls, or may remain unmapped and essentially ignored by downstream informatics pipeline sub-processes. Herein, we describe Amplicon Indel Hunter (AIH), a novel large (>5-bp) indel detection method that is reference genome independent and highly sensitive for the identification of somatic indels in amplicon-based, paired-end, next-generation sequencing data. We validated the algorithm for sensitivity and specificity using a set of clinical cancer samples with Clinical Laboratory Improvement Amendment-confirmed indels as well as in silico mutated data sets. The AIH detected 100% of tested large indels with relatively higher mutant allele frequencies compared with a variety of traditional aligners, which showed variably reduced sensitivity and specificity by comparison. The AIH especially outperformed alignment-based methods for detection of all tested FLT3 internal tandem duplications up to 102 bp. Because AIH runs in parallel to traditional alignment-based informatics pathways, it can be readily incorporated into nearly any analysis pipeline for somatic mutation detection in paired-end amplicon-based data.

Functional expression, characterization and application of the human S100A4 protein.

Preparations utilizing monoclonal antibodies against S100A4 provide useful tools for functional studies to investigate the clinical applications of the human S100A4 protein. In the present study, human S100A4 protein was expressed in Escherichia coli (E. coli) BL21 (DE3), successfully purified by diethylaminoethyl cellulose anion-exchange chromatography and identified by western blot analysis. Soluble S100A4 bioactivity was confirmed by Transwell migration and invasion assays in the human HeLa cell line. Monoclonal antibodies (mAbs) were generated utilizing the standard hybridoma method and were validated by enzyme-linked immunosorbent assay and western blot analysis. The antibody was then used to examine human gastric carcinoma specimens by immunohistochemistry. Recombinant S100A4 was functionally expressed in E. coli and promoted the migration and invasion of HeLa cells. Four hybridoma cell lines, which secreted mAbs specifically against human S100A4 protein, were obtained. One of the four mAbs, namely 2A12D10B2, recognized human S100A4 as indicated by immunohistochemical staining of human gastric carcinoma specimens and recombinant S100A4 was functionally expressed in E. coli. The mAbs of recombinant S100A4 were suitable for detecting S100A4 expression in human tissues and for investigating the subsequent clinical applications of the protein.