36 results on '"Luo, Yuling"'
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2. DNA Tetrahedra-Based Delivery of MicroRNA-22 to Reduce Depressive Symptoms in Mice.
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Luo, Yuling, Yang, Xiao, Du, Yue, Dou, Yikai, Cui, Weitong, Li, Jiajie, Wei, Jinxue, Ma, Xiaohong, and Lin, Yunfeng
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- 2023
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3. Aspirin-Loaded Cross-Linked Lipoic Acid Nanodrug Prevents Postoperative Tumor Recurrence by Residual Cancer Cell Killing and Inflammatory Microenvironment Improvement.
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Jing, Pei, Luo, Yuling, Chen, Yun, Tan, Jiangbing, Liao, Chunyan, and Zhang, Shiyong
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- 2023
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4. A two-branch encoder-decoder network for image tampering localization.
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Luo, Yuling, Liang, Ce, Qin, Sheng, Liu, Junxiu, Fu, Qiang, and Yang, Su
- Abstract
Tampered images with false information can mislead viewers and pose security issues. Tampering traces in images are difficult to detect. To locate tampering traces effectively, a dual-domain deep-learning-based image tampering localization method based on RGB and frequency stream branches is proposed in this work. The former branch learns and extracts tampered features on the image and content features of the tampered region. The latter branch extracts tampered features from the frequency domain to complement the RGB stream branch. In addition, an attention mechanism is used to integrate the features from both branches at the fusion stage. In the experiments, the F 1 score of the proposed method outperformed those of the baselines on the NIST16 dataset (with a 15.3 % improvement), and the AUC score outperformed those of the baselines on the NIST16 and COVERAGE datasets (improvements of 3.9 % and 4.7 %, respectively). This study provides a beneficial alternative to image tampering localization techniques. • A two-branch encoder-decoder network is proposed for locating tampered regions. • Frequency-aware image decomposition enhances network's ability. • Attention mechanism enhances performance by focusing on tampering features. [ABSTRACT FROM AUTHOR]
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- 2024
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5. A novel automated multiplex immunoassay for the ultrasensitive detection of blood‐based biomarkers for neurodegenerative diseases.
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Ma, Xiao‐Jun, Ariyapala, Ishara, Alganem, Khaled, Hao, Qinyu, Feng, Wei, Zhang, Bingqing, Blennow, Kaj, Luo, Yuling, and Zetterberg, Henrik
- Abstract
Background: Efforts to identify accurate blood biomarkers for neurodegenerative disease hallmarks have been hampered by the lack of a proteomic tool that has the required sensitivity to detect very low concentrations of brain‐derived proteins in plasma or serum and the ability to multiplex many analytes in a single assay. Here we evaluated NULISA™, a recently developed novel immunoassay with attomolar level sensitivity and high multiplex capability1, for its ability to detect serum biomarkers associated with Alzheimer's disease. Method: Serum samples from 31 Alzheimer's disease (AD) patients and 31 non‐AD controls were analyzed using NULISA with a 200plex inflammation panel targeting a broad spectrum of inflammation‐related cytokines, chemokines and other proteins. Five samples failed quality control and were excluded from further analysis, resulting in 28 AD and 29 control samples in the final dataset. Linear model analysis, including age and sex as covariates, was performed for each target for differential expression between AD patients and controls. Result: Two significant proteins were identified. The level of glial fibrillary acidic protein (GFAP) was 1.84‐fold higher in AD patients (adjusted p‐value = 0.00014), whereas the level of S100 calcium‐binding protein A12 (S100A12) was 2‐fold higher in non‐AD controls (adjusted p‐value = 0.031). GFAP serum levels correlated strongly with total tau (Spearman rho = 0.69, p = 1.7e‐09), p‐tau181 (rho = 0.69, p = 1.7e‐09) and amyloid beta 42 (rho = ‐0.71, p = 4.7e‐10) levels in cerebrospinal fluid (CSF). S100A12 levels also correlated significantly with total tau (rho = ‐0.39, p = 0.0031), p‐tau181 (rho = ‐0.36, p = 0.0053) and amyloid beta 42 (rho = 0.38, p = 0.0067) CSF levels. Receiver operating characteristic analysis demonstrated that GFAP strongly discriminated AD from controls with an area under the curve (AUC) of 0.93 (95% CI 0.87‐1.0), and S100A12 also exhibited good discrimination with an AUC of 0.72 (95% CI 0.59‐0.86). Conclusion: This exploratory study identified serum GFAP levels as a strong discriminatory biomarker for AD, consistent with previous studies. S100A12 also demonstrated significant associations with AD and CSF biomarkers and thus warrants further study. NULISA holds great promise for the discovery and validation of blood‐based biomarkers for the early detection and monitoring of neurodegenerative diseases. 1. Feng, W. et al. bioRxiv 2023.04.09.536130 (2023) https://doi.org/10.1101/2023.04.09.536130. [ABSTRACT FROM AUTHOR]
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- 2023
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6. Effects of gut microbiota and probiotics on Alzheimer’s disease
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Guo, Libing, Xu, Jiaxin, Du, Yunhua, Wu, Weibo, Nie, Wenjing, Zhang, Dongliang, Luo, Yuling, Lu, Huixian, Lei, Ming, Xiao, Songhua, and Liu, Jun
- Abstract
Alzheimer’s disease (AD) is a progressive neurodegenerative disease with high morbidity, disability, and fatality rate, significantly increasing the global burden of public health. The failure in drug discovery over the past decades has stressed the urgency and importance of seeking new perspectives. Recently, gut microbiome (GM), with the ability to communicate with the brain bidirectionally through the microbiome–gut–brain axis, has attracted much attention in AD-related studies, owing to their strong associations with amyloids, systematic and focal inflammation, impairment of vascular homeostasis and gut barrier, mitochondrial dysfunction, etc., making the regulation of GM, specifically supplementation of probiotics a promising candidate for AD treatment. This article aims to review the leading-edge knowledge concerning potential roles of GM in AD pathogenesis and of probiotics in its treatment and prevention.
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- 2021
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7. Melatonin reduces radiation damage in inner ear
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Chen, Ting, Luo, Yuling, Li, Qi, Yang, Chen, Yuan, Yixin, Peng, Jinhao, Ban, Molu, Liang, Yong, and Zhang, Wei
- Abstract
The purpose of this study was to use a murine model to determine if melatonin can protect the inner ear from radiation-induced damage. A total of 81 4-week-old Balb/c mice were randomly divided into five groups: control group; 50 mg/kg melatonin group; 5 mg/kg melatonin+radiotherapy group; 50 mg/kg melatonin+radiotherapy group; radiotherapy group. The radiotherapy groups received 16 Gy irradiation and melatonin was administered by intraperitoneal injection 30 min before radiotherapy. On days 3 and 7 after irradiation the function of outer hair cells was determined by auditory brainstem response (ABR) and distortion product otoacoustic emissions (DPOAEs) testing, pathological changes of inner ear cells were observed by light microscopy, and the expression of prestin mRNA was determined. ABR thresholds were increased and wave I latencies were extended after radiotherapy; however, the increases were lower in the groups that received melatonin (P< 0.05). DPOAEs showed radiotherapy-induced hearing loss at 8–12 kHz, and hearing loss was greater on day 7 than day 3. However, hearing loss was less in the melatonin groups (P< 0.05). Histopathological examination showed irradiation resulted in breaks and distortion of the cochlear basement membrane, disruption of the stria vascularis, and swelling of outer hair cells. Melatonin reduced these changes. Radiotherapy upregulated prestin mRNA expression. Radiotherapy-induced upregulation of prestin was decreased in the melatonin groups (P< 0.05), and the decrease was greater in the 50 mg/kg melatonin group (P< 0.05). Melatonin protects against radiation-induced cochlear damage by reducing damage to outer hair cells.
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- 2021
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8. A new proposal of utilizing intraoperative electron radiation therapy on the surface of liver to prevent postoperative liver metastasis of pancreatic cancer.
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Dong, Yiting, Song, Zikuan, Luo, Yuling, and Ma, Xuelei
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LIVER metastasis ,PANCREATIC cancer ,INTRAOPERATIVE radiotherapy ,RADIOTHERAPY ,METASTASIS ,MARGINAL distributions ,ELECTRON distribution - Abstract
Pancreatic cancer is a lethal cancer with high rate of liver metastasis worldwide, whereas its treatment choices are limited to a large extent. The limitation of current therapeutic strategies calls for an effective approach which can lower the postoperative liver metastasis rate in order to improve the overall prognosis and survival rate. Comprehensively considering the basic knowledge and clinical practice of tumor treatment worldwide, we proposed three points of hypotheses. Basically, the existing evidences indicated that tumor cells shedding from pancreatic cancer localized in the marginal liver preferentially through the Portal vein. Then, the percentage depth dose distribution of electron radiation is consistent with the marginal distribution of liver metastasis from pancreatic cancer. Based on the characteristics of liver metastasis of pancreatic cancer and the percentage depth dose of electron radiation, we provide a new propose of preventing postoperative liver metastasis in a way of prophylactic intraoperative electron radiation therapy on the surface of liver. Intraoperative electron radiation is relatively easy to control radiation dose and treatment area under direct vision, effectively inhibiting the metastasis and growth of cancer cells and preventing further deterioration of pancreatic cancer patients' condition. Therefore, this hypothesis has an important clinical significance for postoperative rehabilitation and improvement of patients' survival. [ABSTRACT FROM AUTHOR]
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- 2019
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9. NCS-Mediated Direct C(sp3)–H Oxygenation of 2-Methylindoles Using Water as the Oxygen Source
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Zhang, Fan, Luo, Yuling, Liu, Xiaoyi, Liu, Yaoyao, and Xu, Jun
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In continuation of our research interest in the green oxidation of indoles, we further explore the direct oxidation of 2-methylindoles to 2-formyl indoles promoted by NCS and associated with H2O as the oxygen source. This methodology was demonstrated to be a robust protocol consisting of chlorination, SN2′, and oxidation processes, and presents a reasonably broad substrate scope and excellent functional group tolerance, thus enabling the preparation of high added-value versatile building blocks susceptible to further functionalization.
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- 2024
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10. Homeobox B4 optimizes the therapeutic effect of bone marrow mesenchymal stem cells on endotoxin-associated acute lung injury in rats
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Lin, Shan, Luo, Yuling, Mao, Xueyan, He, Wanmei, Xu, Caixia, and Zeng, Mian
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Alveolar capillary endothelial cell (EC) injury has a pivotal role in driving acute respiratory distress syndrome (ARDS) progression and maintaining endothelial homeostasis. A previous ex vivo study revealed that overexpression of homeobox B4 (HOXB4) in bone marrow mesenchymal stem cells (BMSCs) enhanced protection against lipopolysaccharide (LPS)-induced EC injury by activating the Wnt/β-catenin pathway. This in vivo study was performed to verify whether BMSCs overexpressing HOXB4 exert similar protective effects on LPS-induced acute lung injury (ALI) in an animal model.
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- 2024
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11. DNA Tetrahedra-Based Delivery of MicroRNA-22 to Reduce Depressive Symptoms in Mice
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Luo, Yuling, Yang, Xiao, Du, Yue, Dou, Yikai, Cui, Weitong, Li, Jiajie, Wei, Jinxue, Ma, Xiaohong, and Lin, Yunfeng
- Abstract
Major depressive disorder (MDD) is a common illness with an increasing lifetime prevalence. Thus, an increasing number of studies have investigated the association between MDD and microRNAs (miRNAs), which are a novel approach for treating depression. However, the therapeutic potential of miRNA-based strategies has several limitations. To overcome these limitations, DNA tetrahedra (TDNs) have been used as piggyback materials. In this study, we successfully used TDNs as carriers of miRNA-22-3p (miR-22-3p) and synthesized a novel DNA nanocomplex (TDN-miR-22-3p), which was used in a lipopolysaccharide (LPS)-induced depression cell model. The results suggest that miR-22-3p may regulate inflammation by regulating phosphatase and tensin homologue (PTEN), an important regulatory molecule in the PI3K/AKT pathway, and downregulating the expression of NLRP3. We further validated the role of TDN-miR-22-3p in vivo using an LPS-induced animal model of depression. The results indicate that it ameliorated depression-like behavior and attenuated the expression of inflammation-related factors in mice. This study demonstrates the establishment of a straightforward and efficacious miRNA delivery system and the potential of TDNs as therapeutic vectors and tools for mechanistic studies. To the best of our knowledge, this is the first study to use TDNs in combination with miRNAs to treat depression.
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- 2023
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12. Aspirin-Loaded Cross-Linked Lipoic Acid Nanodrug Prevents Postoperative Tumor Recurrence by Residual Cancer Cell Killing and Inflammatory Microenvironment Improvement
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Jing, Pei, Luo, Yuling, Chen, Yun, Tan, Jiangbing, Liao, Chunyan, and Zhang, Shiyong
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In addition to residual cancer cells, the surgery resection-induced hyperinflammatory microenvironment is a key factor that leads to postsurgical cancer recurrence. Herein, we developed a dual-functional nanodrug Asp@cLANVs for postsurgical recurrence inhibition by loading the classical anti-inflammatory drug aspirin (Asp) into cross-linked lipoic acid nanovesicles (cLANVs). The Asp@cLANVs can not only kill residual cancer cells at the doses comparable to common cytotoxic drugs by synergistic interaction between Asp and cLANVs, but also improve the postsurgical inflammatory microenvironment by their strongly synergistic anti-inflammation activity between Asp and cLANVs. Using mice bearing partially removed NCI-H460 tumors, we found that Asp@cLANVs gave a much lower recurrence rate (33.3%) compared with the first-line cytotoxic drug cisplatin (100%), and no mice died for at least 60 days after Asp@cLANV treatment while no mouse survived beyond day 43 in the cisplatin group. This dual-functional nanodrug constructs the first example that combines residual cancer cell killing and postoperative inflammation microenvironment improvement to suppress postsurgical cancer recurrence.
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- 2023
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13. Fluorescence examination and photodynamic therapy of facial squamous cell carcinoma—A case report.
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Zhang, Xiaona, Chen, Jinzhang, Luo, Yuling, Zhang, Lanying, Luo, Rongcheng, and Li, Libo
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CANCER treatment ,SQUAMOUS cell carcinoma ,FLUORESCENCE ,PHOTOCHEMOTHERAPY ,AMINOLEVULINIC acid ,FEASIBILITY studies ,PHOTOSENSITIZERS ,DRUG administration - Abstract
Summary: Background and objective: To evaluate the usefulness and feasibility of fluorescence examination and photodynamic therapy (PDT) for facial squamous cell carcinoma. Methods: A 68-year-old male patient underwent 3 courses of PDT. The first treatment was carried out using topically applied 5-aminolevulinic acid (ALA) to the affected area (5.0cm×2.5cm) 3h before light irradiation. ALA/PpIX-mediated fluorescence was used to visualize the lesion and its margin. The lesion was illuminated with a 630nm laser at 120J/cm
2 . Thirty days later, the residual lesion received the second treatment at the same light dose after topical use of ALA. Despite the tumor was treated with two courses of PDT, The lesion was not cleared completely yet. At last we used topical ALA and intravenous injection of HiPorfin, with the same light dose, then the lesion was cured and the patient was tumor-free followed up for over 15 months. Results: ALA/PpIX-mediated fluorescence visualized the lesion location. Complete cure was achieved after the third course of ALA/Hiporfin PDT. During the 15 months of followed up, no recurrence was noticed. Conclusion: ALA assisted fluorescence examination can be a useful tool to visualize the malignant lesion and determine its margin. ALA-PDT is effective for superficial lesions, but for thicker and deeper lesions, systemic administration of photosensitizer is indispensable. [Copyright &y& Elsevier]- Published
- 2012
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14. Application of early nursing intervention in high risk population of cervical cancer in Guangzhou City.
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Zhou Xiaoping, Luo Yuling, and Li Juan
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- 2016
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15. Papillary Squamous Cell Carcinoma of the Head and Neck
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Mehrad, Mitra, Carpenter, Danielle H., Chernock, Rebecca D., Wang, Hongwei, Ma, Xiao-Jun, Luo, Yuling, Luo, Jingqin, Lewis, James S., and El-Mofty, Samir K.
- Abstract
A relationship between human papillomavirus (HPV) infection and papillary squamous cell carcinoma (PSCC) has been suggested. However, to date, no studies have thoroughly and directly evaluated for transcriptional activity of the virus or the clinicopathologic significance of HPV-positive PSCC. Forty-eight cases of PSCC were retrieved from our surgical pathology database and were reviewed by 4 study pathologists, with tumors defined as SCC with a significant component of papillary growth in the tumor. Immunohistochemical analysis for p16 and p53 was performed. Overexpression of p16 was used as a surrogate marker of transcriptionally active HPV. Transcriptional activity was also directly evaluated using RNA in situ hybridization to detect high-risk HPV E6E7 mRNA. Clinical follow-up data were obtained by chart review. Seven cases were located in the oral cavity, 19 in the oropharynx, and 22 in the larynx. Two morphologic types of PSCC were identified: keratinizing type, in which the epithelial cells showed a maturation trend with minimal surface parakeratin, and nonkeratinizing type, in which the papillae were completely covered by immature basaloid cells. Transcriptionally active HPV was present in 23 of 43 (53.4) tumors. The majority of tumors harboring transcriptionally active HPV arose in the oropharynx, showed nonkeratinizing morphology, were p16 positive, and p53 negative. Transcriptionally active HPV was also present in many laryngeal and oral cavity PSCCs. Overall survival, disease-specific survival, and disease-free survival were favorable and did not significantly differ by anatomic subsite. However, HPV-related tumors showed a trend toward better survival.
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- 2013
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16. Automated Quantitative RNA in SituHybridization for Resolution of Equivocal and Heterogeneous ERBB2(HER2) Status in Invasive Breast Carcinoma
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Wang, Zhen, Portier, Bryce P., Gruver, Aaron M., Bui, Son, Wang, Hongwei, Su, Nan, Vo, Hong-Thuy, Ma, Xiao-Jun, Luo, Yuling, Budd, G. Thomas, and Tubbs, Raymond R.
- Abstract
Patient management based on HER2status in breast carcinoma is an archetypical example of personalized medicine but remains hampered by equivocal testing and intratumoral heterogeneity. We developed a fully automated, quantitative, bright-field in situhybridization technique (RNAscope), applied it to quantify single-cell HER2mRNA levels in 132 invasive breast carcinomas, and compared the results with those by real-time quantitative PCR (qPCR) and Food and Drug Administration–approved methods, including fluorescence in situhybridization (FISH), IHC, chromogenic in situhybridization, and dual in situhybridization. Both RNAscope and qPCR were 97.3% concordant with FISH in cases in which FISH results were unequivocal. RNAscope was superior to qPCR in cases with intratumoral heterogeneity or equivocal FISH results. This novel assay may enable ultimate HER2status resolution as a reflex test for current testing algorithms. Quantitative in situRNA measurement at the single-cell level may be broadly applicable in companion diagnostic applications.
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- 2013
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17. Detection of Transcriptionally Active High-risk HPV in Patients With Head and Neck Squamous Cell Carcinoma as Visualized by a Novel E6E7 mRNA In Situ Hybridization Method
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Bishop, Justin A., Ma, Xiao-Jun, Wang, Hongwei, Luo, Yuling, Illei, Peter B., Begum, Shanaz, Taube, Janis M., Koch, Wayne M., and Westra, William H.
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Evidence for transcriptional activation of the viral oncoproteins E6 and E7 is regarded as the gold standard for the presence of clinically relevant human papillomavirus (HPV), but detection of E6E7 mRNA requires RNA extraction and polymerase chain reaction amplification—a challenging technique that is restricted to the research laboratory. The development of RNA in situ hybridization (ISH) probes complementary to E6E7 mRNA permits direct visualization of viral transcripts in routinely processed tissues and has opened the door for accurate HPV detection in the clinical care setting. Tissue microarrays containing 282 head and neck squamous cell carcinomas from various anatomic subsites were tested for the presence of HPV using p16 immunohistochemistry, HPV DNA ISH, and an RNA ISH assay (RNAscope) targeting high-risk HPV E6E7 mRNA transcripts. The E6E7 mRNA assay was also used to test an additional 25 oropharyngeal carcinomas in which the HPV status as recorded in the surgical pathology reports was equivocal due to conflicting detection results (ie, p16 positive, DNA ISH negative). By the E6E7 mRNA method, HPV was detected in 49 of 282 (17) HNSCCs including 43 of 77 (56) carcinomas from the oropharynx, 2 of 3 (67) metastatic HNSCCs of an unknown primary site, 2 of 7 (29) carcinomas from the sinonasal tract, and 2 of 195 (1) carcinomas from other head and neck sites. p16 expression was strongly associated with the presence of HPV E6E7 mRNA: 46 of 49 HPV-positive tumors exhibited p16 expression, whereas only 22 of 233 HPV-negative tumors were p16 positive (94 vs. 9, P<0.0001). There was also a high rate of concordance (99) between the E6E7 mRNA method and HPV DNA ISH. For the selected group of discordant HNSCCs (p16HPV DNA−), the presence of E6E7 transcripts was detected in 21 of 25 (84) cases. The E6E7 mRNA method confirmed the presence of transcriptionally active HPV-related HNSCC that has a strong predilection for the oropharynx and is strongly associated with high levels of p16 expression. Testing for HPV E6E7 transcripts by RNA ISH is ideal because it confirms the presence of integrated and transcriptionally active virus, permits visualization of viral transcripts in tissues, and is technically feasible for routine testing in the clinical laboratory.
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- 2012
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18. Partial p16 staining in oropharyngeal squamous cell carcinoma: extent and pattern correlate with human papillomavirus RNA status
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Lewis, James S, Chernock, Rebecca D, Ma, Xiao-Jun, Flanagan, John J, Luo, Yuling, Gao, Ge, Wang, Xiaowei, and El-Mofty, Samir K
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Human papillomavirus (HPV)-related oropharyngeal squamous cell carcinoma has unique biology and better outcomes. p16 immunostaining is used as a surrogate marker for transcriptionally active HPV. Although diffuse staining is generally accepted as positive, the significance of partial staining has not been established, nor has the cutoff for extent of p16 staining that should be used to identify a tumor as HPV-related. From three other large studies utilizing p16 immunohistochemistry, we identified all cases with partial positive staining. The p16-stained slides were reviewed by three study pathologists for staining (nuclear and cytoplasmic) extent (in quartiles), and also for percentage that was confluent (ie, back-to-back cell staining). Tumors were histologically typed (keratinizing, non-keratinizing, or non-keratinizing with maturation) and tested for high-risk HPV by RNA in-situhybridization and reverse-transcriptase PCR. For the 16 cases, there were two 4+(13%), five 3+(31%), six 2+(38%), and three 1+(19%) p16 staining tumors. Extent of staining ranged from 5 to 90% of cells positive with 25% or more confluent staining in 4/16 (25%). Of the 16 (31%) cases, 5 were HPV-related on the basis of RNA in-situhybridization and reverse-transcriptase PCR. All of these cases had >50% p16 staining, 4/5 (80%) had more than 25% confluent staining, and 4/7 (57%) had non-keratinizing histological features. Only one of the p16 1+/2+ tumors was HPV RNA-positive (by reverse-transcriptase PCR only and low level). All 1+/2+ cases were keratinizing type or undifferentiated. By sensitive detection methods, most partial p16-positive squamous cell carcinoma cases with >50% staining harbor transcriptionally active HPV, and most HPV+ tumors have significant amounts of confluent staining. Cases with <50% p16 staining and lacking significant confluent staining rarely harbor HPV. These results support that greater than 75% p16 staining or, alternatively, >50% staining combined with >25% confluent areas, are suitable cutoffs for defining positivity.
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- 2012
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19. Partial p16 staining in oropharyngeal squamous cell carcinoma: extent and pattern correlate with human papillomavirus RNA status
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Lewis, James S, Chernock, Rebecca D, Ma, Xiao-Jun, Flanagan, John J, Luo, Yuling, Gao, Ge, Wang, Xiaowei, and El-Mofty, Samir K
- Abstract
Human papillomavirus (HPV)-related oropharyngeal squamous cell carcinoma has unique biology and better outcomes. p16 immunostaining is used as a surrogate marker for transcriptionally active HPV. Although diffuse staining is generally accepted as positive, the significance of partial staining has not been established, nor has the cutoff for extent of p16 staining that should be used to identify a tumor as HPV-related. From three other large studies utilizing p16 immunohistochemistry, we identified all cases with partial positive staining. The p16-stained slides were reviewed by three study pathologists for staining (nuclear and cytoplasmic) extent (in quartiles), and also for percentage that was confluent (ie, back-to-back cell staining). Tumors were histologically typed (keratinizing, non-keratinizing, or non-keratinizing with maturation) and tested for high-risk HPV by RNA in-situ hybridization and reverse-transcriptase PCR. For the 16 cases, there were two 4+(13%), five 3+(31%), six 2+(38%), and three 1+(19%) p16 staining tumors. Extent of staining ranged from 5 to 90% of cells positive with 25% or more confluent staining in 4/16 (25%). Of the 16 (31%) cases, 5 were HPV-related on the basis of RNA in-situ hybridization and reverse-transcriptase PCR. All of these cases had >50% p16 staining, 4/5 (80%) had more than 25% confluent staining, and 4/7 (57%) had non-keratinizing histological features. Only one of the p16 1+/2+ tumors was HPV RNA-positive (by reverse-transcriptase PCR only and low level). All 1+/2+ cases were keratinizing type or undifferentiated. By sensitive detection methods, most partial p16-positive squamous cell carcinoma cases with >50% staining harbor transcriptionally active HPV, and most HPV+ tumors have significant amounts of confluent staining. Cases with <50% p16 staining and lacking significant confluent staining rarely harbor HPV. These results support that greater than 75% p16 staining or, alternatively, >50% staining combined with >25% confluent areas, are suitable cutoffs for defining positivity.
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- 2012
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20. RNAscope
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Wang, Fay, Flanagan, John, Su, Nan, Wang, Li-Chong, Bui, Son, Nielson, Allissa, Wu, Xingyong, Vo, Hong-Thuy, Ma, Xiao-Jun, and Luo, Yuling
- Abstract
In situanalysis of biomarkers is highly desirable in molecular pathology because it allows the examination of biomarker status within the histopathological context of clinical specimens. Immunohistochemistry and DNA in situhybridization (ISH) are widely used in clinical settings to assess protein and DNA biomarkers, respectively, but clinical use of in situRNA analysis is rare. This disparity is especially notable when considering the abundance of RNA biomarkers discovered through whole-genome expression profiling. This is largely due to the high degree of technical complexity and insufficient sensitivity and specificity of current RNA ISH techniques. Here, we describe RNAscope, a novel RNA ISH technology with a unique probe design strategy that allows simultaneous signal amplification and background suppression to achieve single-molecule visualization while preserving tissue morphology. RNAscope is compatible with routine formalin-fixed, paraffin-embedded tissue specimens and can use either conventional chromogenic dyes for bright-field microscopy or fluorescent dyes for multiplex analysis. Unlike grind-and-bind RNA analysis methods such as real-time RT-PCR, RNAscope brings the benefits of in situanalysis to RNA biomarkers and may enable rapid development of RNA ISH-based molecular diagnostic assays.
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- 2012
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21. Direct Quantification of Gene Expression in Homogenates of Formalin-Fixed, Paraffin-Embedded Tissues
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Yang, Wen, Maqsodi, Botoul, Ma, Yunqing, Bui, Son, Crawford, Kimberly L., McMaster, Gary K., Witney, Frank, and Luo, Yuling
- Abstract
Formalin-fixed, paraffin-embedded (FFPE) tissues represent an important source of archival materials for gene expression profiling. We report here the development of a modified branch DNA assay that allows direct quantification of messenger RNA (mRNA) transcripts in homogenates from FFPE tissue sections without the need for RNA isolation and reverse transcription into cDNA. Formalin fixation essentially has no effect on the branch DNA assay, and RNA degradation only marginally reduces the signal by 2- to 3-fold. Under the same conditions, formalin fixation and RNA degradation greatly reduces real-time reverse transcription PCR (RT-PCR) efficiency, reducing signals by as much as 15- and 1400-fold, respectively. Although both technologies can generate biologically meaningful expression profiles from FFPE human lung tumor specimens, the branch DNA assay is more sensitive than real-time RT-PCR under the conditions tested. Our results therefore suggest that the branch DNA assay is an ideal tool for retrospective analysis of gene expression in archival tissues.
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- 2006
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22. Small Interfering RNA and Gene Expression Analysis Using a Multiplex Branched DNA Assay without RNA Purification
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Zhang, Aiguo, Pastor, Larry, Nguyen, Quan, Luo, Yuling, Yang, Wen, Flagella, Michael, Chavli, Rajesh, Bui, Son, Nguyen, Cung Tuong, Zheng, Zhi, He, Weihai, McMaster, Gary, and Witney, Frank
- Abstract
The authors have developed a novel multiplex detection system that quantitatively measures the expression level of 11 messenger RNAs (mRNAs) directly from cell lysates or tissue homogenates without RNA purification. The system incorporates branched DNA (bDNA) technology from Bayer and a multiplex bead array platform from Luminex. In this study, a 21-nt synthetic small interfering RNA (siRNA; specifically designed to knockdown interleukin-8 [IL-8] expression) was delivered into HeLa cells. Using the multiplex bDNA assay, gene expression levels were measured simultaneously from cell lysates for 11 genes. After treating the HeLa cells for 20 h with phorbol myristate acetate (PMA), IL-8 mRNA levels were induced by almost 50-fold; transfection with 30 nM IL-8-specific siRNA reduced the PMA-induced IL-8 mRNA by 80%. In addition, PMA induced mRNA expression in IL-1α (3-fold) and IL-6 (4-fold); however, the IL-8 siRNA did not affect the expression of either of these 2 cytokine genes, indicating that the siRNA was selective for IL-8 mRNA expression. Three housekeeping genes’ expression levels were measured under all conditions tested. The multiplex bDNA assay provides a powerful tool for quantitative multiplex gene expression analysis directly from cell lysates, which could be extremely valuable for conservation of rare or difficult-to-obtain samples.
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- 2005
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23. Small Interfering RNA and Gene Expression Analysis Using a Multiplex Branched DNA Assay without RNA Purification
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Zhang, Aiguo, Pastor, Larry, Nguyen, Quan, Luo, Yuling, Yang, Wen, Flagella, Michael, Chavli, Rajesh, Bui, Son, Nguyen, Cung Tuong, Zheng, Zhi, He, Weihai, McMaster, Gary, and Witney, Frank
- Abstract
The authors have developed a novel multiplex detection system that quantitatively measures the expression level of 11 messenger RNAs (mRNAs) directly from cell lysates or tissue homogenates without RNA purification. The system incorporates branched DNA (bDNA) technology from Bayer and a multiplex bead array platform from Luminex. In this study, a 21-nt synthetic small interfering RNA (siRNA; specifically designed to knockdown interleukin-8 [IL-8] expression) was delivered into HeLa cells. Using the multiplex bDNA assay, gene expression levels were measured simultaneously from cell lysates for 11 genes. After treating the HeLa cells for 20 h with phorbol myristate acetate (PMA), IL-8 mRNA levels were induced by almost 50-fold; transfection with 30 nM IL-8-specific siRNA reduced the PMA-induced IL-8 mRNA by 80%. In addition, PMA induced mRNA expression in IL-1α (3-fold) and IL-6 (4-fold); however, the IL-8 siRNA did not affect the expression of either of these 2 cytokine genes, indicating that the siRNA was selective for IL-8 mRNA expression. Three housekeeping genes’ expression levels were measured under all conditions tested. The multiplex bDNA assay provides a powerful tool for quantitative multiplex gene expression analysis directly from cell lysates, which could be extremely valuable for conservation of rare or difficult-to-obtain samples. (Journal of Biomolecular Screening2005:549-556)
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- 2005
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24. Temporal specific patterns of semaphorin gene expression in rat brain after kainic acid‐induced status epilepticus
- Author
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Barnes, Gregory, Puranam, Ram S., Luo, Yuling, and McNamara, James O.
- Abstract
Mossy fiber sprouting and other forms of synaptic reorganization may form the basis for a recurrent excitatory network in epileptic foci. Four major classes of axon guidance molecules—the ephrins, netrins, slits, and semaphorins—provide targeting information to outgrowing axons along predetermined pathways during development. These molecules may also play a role in synaptic reorganization in the adult brain and thereby promote epileptogenesis. We studied semaphorin gene expression, as assessed by in situ hybridization, using riboprobes generated from rat cDNA in an adult model of synaptic reorganization, kainic acid (KA)‐induced status epilepticus (SE). Within the first week after KA‐induced SE, semaphorin 3C, a class III semaphorin, mRNA content is decreased in the CA1 area of the hippocampus and is increased in the upper layers of cerebral cortex. Another class III semaphorin, semaphorin 3F, is also decreased in CA1 and CA3 of hippocampus within the first week after KA‐SE. These changes in gene expression are principally confined to neurons. By contrast, there was little change in the semaphorin 4C mRNA content of CA1 neurons at this time. No changes in expression of semaphorin 3A and 4C genes were detected 28 days after KA‐induced SE. Regulation of semaphorin gene expression after KA‐induced SE suggests that neurons may regulate the expression of axonal guidance molecules and thereby contribute to synaptic reorganization after injury of the mature brain. The anatomic locale of the altered semaphorin gene expression may serve as a marker for specific networks undergoing synaptic reorganization in the epileptic brain. Hippocampus 2003;13:1–20. © 2003 Wiley‐Liss, Inc.
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- 2003
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25. Temporal specific patterns of semaphorin gene expression in rat brain after kainic acid-induced status epilepticus
- Author
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Barnes, Gregory, Puranam, Ram S., and Luo, Yuling
- Abstract
Mossy fiber sprouting and other forms of synaptic reorganization may form the basis for a recurrent excitatory network in epileptic foci. Four major classes of axon guidance moleculesthe ephrins, netrins, slits, and semaphorinsprovide targeting information to outgrowing axons along predetermined pathways during development. These molecules may also play a role in synaptic reorganization in the adult brain and thereby promote epileptogenesis. We studied semaphorin gene expression, as assessed by in situ hybridization, using riboprobes generated from rat cDNA in an adult model of synaptic reorganization, kainic acid (KA)-induced status epilepticus (SE). Within the first week after KA-induced SE, semaphorin 3C, a class III semaphorin, mRNA content is decreased in the CA1 area of the hippocampus and is increased in the upper layers of cerebral cortex. Another class III semaphorin, semaphorin 3F, is also decreased in CA1 and CA3 of hippocampus within the first week after KA-SE. These changes in gene expression are principally confined to neurons. By contrast, there was little change in the semaphorin 4C mRNA content of CA1 neurons at this time. No changes in expression of semaphorin 3A and 4C genes were detected 28 days after KA-induced SE. Regulation of semaphorin gene expression after KA-induced SE suggests that neurons may regulate the expression of axonal guidance molecules and thereby contribute to synaptic reorganization after injury of the mature brain. The anatomic locale of the altered semaphorin gene expression may serve as a marker for specific networks undergoing synaptic reorganization in the epileptic brain. Hippocampus 2003;13:120. © 2003 Wiley-Liss, Inc.
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- 2003
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26. SEMA3A regulates developing sensory projections in the chicken spinal cord
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Fu, Susan Y., Sharma, Kamal, Luo, Yuling, Raper, Jonathan A., and Frank, Eric
- Abstract
The present study explores the role of SEMA3A (collapsin-1) in the temporal and spatial regulation of developing sensory projections in the chick spinal cord. During development, SEMA3A mRNA (SEMA3A) is first expressed throughout the spinal gray matter, but disappears from the dorsal region when small caliber (trkA+) sensory axon collaterals first grow into the dorsal horn. In explant cultures of spinal cord segments with attached sensory ganglia, the spatial extent of SEMA3A expression varied in different explants, but in each case the growth of trkA+ sensory collaterals was largely excluded from areas of SEMA3A expression. To test if SEMA3A had a direct effect on sensory axon growth, we injected recombinant protein into the explants before placing them in culture. Increased levels of SEMA3A substantially reduced the ingrowth of trkA+ axons, whereas trkC+ axon collaterals were not affected. Consistent with the insensitivity of trkC+ collaterals to SEMA3A, these collaterals did not express neuropilin-1, a receptor for SEMA3A. The inhibitory effects of SEMA3A on trkA+ axons within the spinal cord suggests that the fall in SEMA3A expression in the dorsal horn may contribute to the initiation of growth of these axons into gray matter. In addition, the observation that trkA+ axons frequently grew close to but rarely over areas of SEMA3A expression suggests that semaphorin may act principally as a short-range guidance cue within the spinal cord. © 2000 John Wiley & Sons, Inc. J Neurobiol 45: 227236, 2000
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- 2000
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27. The Distribution of Collapsin-1 mRNA in the Developing Chick Nervous System
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Shepherd, Iain, Luo, Yuling, Raper, Jonathan A., and Chang, Susannah
- Abstract
Collapsin-1 is a secreted glycoprotein that inhibits the extension of specific growth conesin vitro.It has been hypothesized to serve as a repulsive guidance cue for extending growth conesin vivo.Here we report the distribution of collapsin-1 message as demonstrated byin situhybridization using digoxigenin-labeled RNA probes in wholemounts and tissue sections. In the early chick brain collapsin-1 is expressed in specific regions of the retina, the olfactory bulb, and the diencephalon. In the hindbrain collapsin-1 is first expressed in rhombomere 5 and later in bilaterally symmetric rostrocaudal stripes. Collapsin-1 is expressed in high levels in the ventral horn of the spinal cord and in ventricular stripes that extend rostrally to the hindbrain. In the periphery, collapsin-1 is expressed in the dermamyotome and in ectoderm and epidermis. Based on collapsin's expression patterns we tested axons extending from explants of ventral spinal cord and olfactory bulb for sensitivity to collapsin and show that the former are sensitive to collapsin wheras the latter are not. The distribution of collapsin mRNA is consistent with it playing a role in the organization of sensory axonal projections within the spinal cord and skin.
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- 1996
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28. Human Semaphorin K1 Is Glycosylphosphatidylinositol-linked and Defines a New Subfamily of Viral-related Semaphorins*
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Xu, Xiaomei, Ng, Sheldon, Wu, Zhi-Liang, Nguyen, Dat, Homburger, Sheila, Seidel-Dugan, Cynthia, Ebens, Allen, and Luo, Yuling
- Abstract
The semaphorin family contains a large number of secreted and transmembrane proteins, some of which are known to act as repulsive axon guidance cues during development or to be involved in immune function. We report here on the identification of semaphorin K1 (sema K1), the first semaphorin known to be associated with cell surfaces via a glycosylphosphatidylinositol linkage. Sema K1 is highly homologous to a viral semaphorin and can interact with specific immune cells, suggesting that like its viral counterpart, sema K1 could play an important role in regulating immune function. Sema K1 does not bind to neuropilin-1 or neuropilin-2, the two receptors implicated in mediating the repulsive action of several secreted semaphorins, and thus it likely acts through a novel receptor. In contrast to most previously described semaphorins, sema K1 is only weakly expressed during development but is present at high levels in postnatal and adult tissues, particularly brain and spinal cord.
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- 1998
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29. A sensory axon repellent secreted from ventral spinal cord explants is neutralized by antibodies raised against collapsin-1
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Shepherd, Iain T., Luo, Yuling, Lefcort, Frances, Reichardt, Louis F., and Raper, Jonathan A.
- Abstract
During embryogenesis, different subclasses of sensory neurons extend central projections to specific locations in the spinal cord. Muscle and cutaneous afferents initially project to the same location in the dorsal cord. Later, specific muscle afferents leave other afferents behind and project into the ventral cord. Previous studies have shown that ventral spinal cord explants secrete a repellent for sensory neurites. We now find that antibodies to collapsin-1 neutralize this repellent activity. Additional data suggest that all afferents respond to collapsin-1 when they are first confined to the dorsal cord, but that ventrally projecting muscle afferents become collapsin-1 insensitive as they project into the ventral cord. Our results suggest that the transient dorsal expression of collapsin-1 prevents all efferents from entering the cord early and sustained ventral expression prevents dorsally terminating afferents from entering the ventral cord later.
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- 1997
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30. Detection of ALK mRNA Transcripts in Liquid-based NSCLC ThinPrep Cytology Slides.
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Reynolds, Jordan, Minca, Eugen, Sanda, Sherrie, Lanigan, Christopher, Kim, Daniel, Bui, Sui, Su, Nan, Ma, Xiao-Jun, Luo, Yuling, Wang, Kaixin, and Wang, Zhen
- Published
- 2014
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31. Detection of ALK mRNA Transcripts in Liquid-based NSCLC ThinPrep Cytology Slides
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Reynolds, Jordan, Minca, Eugen, Sanda, Sherrie, Lanigan, Christopher, Kim, Daniel, Bui, Sui, Su, Nan, Ma, Xiao-Jun, Luo, Yuling, Wang, Kaixin, and Wang, Zhen
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- 2014
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32. Identification of a Disease-Defining Gene Fusion in Epithelioid Hemangioendothelioma
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Tanas, Munir R., Sboner, Andrea, Oliveira, Andre M., Erickson-Johnson, Michele R., Hespelt, Jessica, Hanwright, Philip J., Flanagan, John, Luo, Yuling, Fenwick, Kerry, Natrajan, Rachael, Mitsopoulos, Costas, Zvelebil, Marketa, Hoch, Benjamin L., Weiss, Sharon W., Debiec-Rychter, Maria, Sciot, Raf, West, Rob B., Lazar, Alexander J., Ashworth, Alan, Reis-Filho, Jorge S., Lord, Christopher J., Gerstein, Mark B., Rubin, Mark A., and Rubin, Brian P.
- Abstract
A newly identified gene fusion defines the vascular cancer epithelioid hemangioendothelioma and encodes a chimeric transcription factor.
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- 2011
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33. The Effect of Iodine-131-Rituximab on B Cell Lymphoma In Vitro and In Vivo.
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Luo, Rongcheng, Zuo, Qiang, Wei, Li, Liang, Wangjun, Zhen, Dayong, Luo, Yuling, Yan, Xiao, and Yang, Yang
- Abstract
The purpose of this study was to investigate the cell specific cytotoxic effect of Iodine-131 Rituximab on CD20-positive B cell lymphoma in vitro and on Raji cell tumors grown in vivo. Rituximab was labeled with Iodine -131 by the iodogen method. Cultured Raji cells or the nude mice bearing Raji tumors were treated with various concentrations of Iodine-131-Rituximab or Iodine-131 alone or Rituximab alone. The results showed that The lethal effect was found on Raji cells treated with Iodine-131-Rituximab in a dose-dependent manner; The proliferation rate of Raji cells was significantly lower in cells treated with Iodine-131-Rituximab, as compared to the cells treated with Iodine-131or Rituximab alone (P<0.05); Tumor inhibition was found to be greatest in the mice treated with Iodine-131-Rituximab through intratumor injection, as compared with Iodine-131-Rituximab i.p. injection or Rituximab alone (p<0.05). We conclude that Iodine-131-Rituximab specifically inhibits the growth of Raji tumor cells in vitro and in vivo. Iodine-131-Rituximab is a promising agent for radioimmunotherapy that targets CD20-positive B cell lymphoma.
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- 2007
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34. The Effect of Iodine-131-Rituximab on B Cell Lymphoma In Vitro and In Vivo.
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Luo, Rongcheng, Zuo, Qiang, Wei, Li, Liang, Wangjun, Zhen, Dayong, Luo, Yuling, Yan, Xiao, and Yang, Yang
- Abstract
The purpose of this study was to investigate the cell specific cytotoxic effect of Iodine-131 Rituximab on CD20-positive B cell lymphoma in vitroand on Raji cell tumors grown in vivo.Rituximab was labeled with Iodine -131 by the iodogen method. Cultured Raji cells or the nude mice bearing Raji tumors were treated with various concentrations of Iodine-131-Rituximab or Iodine-131 alone or Rituximab alone. The results showed that
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- 2007
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35. Sensitive and Quantitative Measurement of Gene Expression Directly from Peripheral Whole Blood, without RNA Isolation and Target Amplification.
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Zheng, Zhi, Luo, Yuling, and McMaster, Gary
- Abstract
Peripheral blood gene expression analysis is increasingly used for diagnosis and prognosis of hematological diseases, as well as for surrogate biomarker discovery in a wide range of non-hematological disorders. However, a critical issue concerning the potential clinical application of these research findings relates to the validity and reproducibility of the blood mRNA quantitation results. Current gene expression technologies often depend on multiple steps: 1) blood isolation; 2) RNA purification and 3) subsequent enzymatic reactions, which affect the accuracy and consistency of results. We describe an assay to measure single- and multi-plexed gene expression directly from whole blood without RNA purification and target amplification. The hybridization-based assay uses branched DNA signal amplification technology with a whole blood lysis protocol that preserves the RNA integrity. The assay is sensitive enough to quantitatively measure genes expressed at low levels in a minority of cells from less than 30ul of whole blood. The coefficients of variations are less than 10% and the dynamic range is 3–4 logs for both singleplex and multiplex formats. The assay signals are several times higher than purified RNA from an equivalent amount of blood. Blood proteins, genomic DNA and reticulocyte mRNAs do not interfere with the assay. The assays are compatible with common anticoagulants and Paxgene treated samples. However, we found the Paxgene induced expression of antiapoptotic genes during processing of the whole blood. We used the multiplex assay to evaluate the impact of common blood processing on the expression of a panel of 30 cytokine and apoptosis genes known to be sensitive to ex vivo purtabation. Minimal impact was found with RBC lysis, followed by whole blood RNA extraction, PBMC isolation and Paxgene stabilization. The lowest correlation of expression was found between RNA extracted from whole blood and RNA extracted from same blood treated with Paxgene (R square=0.77). We believe this assay will contribute to fundamental and therapeutic applications where quantitative gene expression and/or throughput are required.
- Published
- 2005
- Full Text
- View/download PDF
36. Sensitive and Quantitative Measurement of Gene Expression Directly from Peripheral Whole Blood, without RNA Isolation and Target Amplification.
- Author
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Zheng, Zhi, Luo, Yuling, and McMaster, Gary
- Abstract
Peripheral blood gene expression analysis is increasingly used for diagnosis and prognosis of hematological diseases, as well as for surrogate biomarker discovery in a wide range of non-hematological disorders. However, a critical issue concerning the potential clinical application of these research findings relates to the validity and reproducibility of the blood mRNA quantitation results. Current gene expression technologies often depend on multiple steps: 1) blood isolation; 2) RNA purification and 3) subsequent enzymatic reactions, which affect the accuracy and consistency of results. We describe an assay to measure single- and multi-plexed gene expression directly from whole blood without RNA purification and target amplification. The hybridization-based assay uses branched DNA signal amplification technology with a whole blood lysis protocol that preserves the RNA integrity. The assay is sensitive enough to quantitatively measure genes expressed at low levels in a minority of cells from less than 30ul of whole blood. The coefficients of variations are less than 10% and the dynamic range is 3–4 logs for both singleplex and multiplex formats. The assay signals are several times higher than purified RNA from an equivalent amount of blood. Blood proteins, genomic DNA and reticulocyte mRNAs do not interfere with the assay. The assays are compatible with common anticoagulants and Paxgene treated samples. However, we found the Paxgene induced expression of antiapoptotic genes during processing of the whole blood. We used the multiplex assay to evaluate the impact of common blood processing on the expression of a panel of 30 cytokine and apoptosis genes known to be sensitive to ex vivo purtabation. Minimal impact was found with RBC lysis, followed by whole blood RNA extraction, PBMC isolation and Paxgene stabilization. The lowest correlation of expression was found between RNA extracted from whole blood and RNA extracted from same blood treated with Paxgene (R square=0.77). We believe this assay will contribute to fundamental and therapeutic applications where quantitative gene expression and/or throughput are required.
- Published
- 2005
- Full Text
- View/download PDF
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