ABSTRACT
Triple-negative breast cancer (TNBC) has high heterogeneity, poor prognosis, and limited treatment success. Recently, an immunohistochemistry-based surrogate classification for the "Fudan University Shanghai Cancer Center (FUSCC) subtyping" has been developed and is considered more suitable for clinical application. Seventy-one paraffin-embedded sections of surgically resected TNBC were classified into four molecular subtypes using the IHC-based surrogate classification. Genomic analysis was performed by targeted next-generation sequencing and the specificity of the subtypes was explored by bioinformatics, including survival analysis, multivariate Cox regression, pathway enrichment, Pyclone analysis, mutational signature analysis and PHIAL analysis. AKT1 and BRCA1 mutations were identified as independent prognostic factors in TNBC. TNBC molecular subtypes encompass distinct genomic landscapes that show specific heterogeneities. The luminal androgen receptor (LAR) subtype was associated with mutations in PIK3CA and PI3K pathways, which are potentially sensitive to PI3K pathway inhibitors. The basal-like immune-suppressed (BLIS) subtype was characterized by high genomic instability and the specific possession of signature 19 while patients in the immunomodulatory (IM) subtype belonged to the PD-L1 ≥ 1% subgroup with enrichment in Notch signaling, suggesting a possible benefit of immune checkpoint inhibitors and Notch inhibitors. Moreover, mesenchymal-like (MES) tumors displayed enrichment in the receptor tyrosine kinase (RTK)-RAS pathway and potential sensitivity to RTK pathway inhibitors. The findings suggest potential treatment targets and prognostic factors, indicating the possibility of TNBC stratified therapy in the future.
Subject(s)
Mutation , Triple Negative Breast Neoplasms , Humans , Triple Negative Breast Neoplasms/genetics , Triple Negative Breast Neoplasms/drug therapy , Triple Negative Breast Neoplasms/pathology , Female , Middle Aged , Proto-Oncogene Proteins c-akt/metabolism , Proto-Oncogene Proteins c-akt/genetics , Prognosis , Class I Phosphatidylinositol 3-Kinases/genetics , Class I Phosphatidylinositol 3-Kinases/metabolism , Genomics/methods , BRCA1 Protein/genetics , Adult , Biomarkers, Tumor/genetics , Aged , High-Throughput Nucleotide Sequencing , B7-H1 Antigen/genetics , B7-H1 Antigen/metabolismABSTRACT
Increasing evidence has demonstrated the oncogenic roles of long non-coding RNA (lncRNA) hepatocellular carcinoma (HCC)-associated long non-coding RNA (HANR) in the development of HCC and lung cancer; however, the involvement of HANR in triple-negative breast cancer (TNBC) remains largely unknown. Our results demonstrated the significant overexpression of HANR in TNBC tissues and cells. Higher HANR levels significantly correlated with the poorer phenotypes in patients with TNBC. HANR down-regulation inhibited the proliferation and cell cycle progression and increased the apoptosis of TNBC cells. Mechanistically, immunoprecipitation-mass spectrometry revealed hexokinase II (HK2) as a direct binding target of HANR. HANR binds to and stabilizes HK2 through the proteasomal pathway. Consistent with the important role of HK2 in cancer cells, HANR depletion represses the glucose absorbance and lactate secretion, thus reprogramming the metabolism of TNBC cells. An in vivo xenograft model also demonstrated that HANR promoted tumor growth and aerobic glycolysis. This study reveals the role of HANR in modulating the glycolysis in TNBC cells by regulating HK2 stability, suggesting that HANR is a potential drug target for TNBC.
ABSTRACT
Circulating miRNAs (cirmiRNAs) can be packaged into the exosomes, participating in intercellular communication, which affects the malignant progression and therapy resistance of triple-negative breast cancer (TNBC). Currently, immune checkpoint inhibitors that regulate T-cell function, especially antibodies against programmed cell death 1 (PD-1) or its ligand PD-L1, are emerging as new promising therapy for TNBC patients. However, only very limited patients showed complete or partial response to anti-PD-1 treatment. Dysfunction of CD8+ T cells is one of the key reasons for the immune escape of TNBC. The regulation of exosome-derived cirmiRNAs on CD8+ T cells in TNBC deserves more investigation. Here, the cirmiR-20a-5p level was significantly upregulated in the plasma of TNBC patients and culture supernatant of TNBC cells. High abundance of cirmiR-20a-5p was correlated with a worse prognosis of TNBC. cirmiR-20a-5p was secreted in the form of exosomes by TNBC cells. Exosomal cirmiR-20a-5p was internalized into CD8+ T cells and resulted into the dysfunction of CD8+ T. A mechanism study uncovered that cirmiR-20a-5p targeted the nuclear protein ataxia-telangiectasia (NPAT) and decreased NPAT expression in CD8+ T cells. An in vivo xenograft mouse model showed that cirmiR-20a-5p conferred TNBC to anti-PD-1 treatment resistance. Collectively, these findings indicated that cirmiR-20a-5p released by TNBC cells via exosome promotes cancer cell growth and leads to the immunosuppression by inducing CD8+ T cell dysfunction. This study suggests that targeting cirmiR-20a-5p might be a novel strategy for overcoming the resistance of TNBC to anti-PD-1 immunotherapy.
Subject(s)
MicroRNAs , Triple Negative Breast Neoplasms , Humans , Animals , Mice , MicroRNAs/genetics , MicroRNAs/metabolism , Triple Negative Breast Neoplasms/drug therapy , Triple Negative Breast Neoplasms/genetics , Triple Negative Breast Neoplasms/metabolism , CD8-Positive T-Lymphocytes/metabolism , Cell Line, Tumor , Cell Proliferation/physiology , B7-H1 Antigen/geneticsABSTRACT
MOTIVATION: Longer reads produced by PacBio or Oxford Nanopore sequencers could more frequently span the breakpoints of structural variations (SVs) than shorter reads. Therefore, existing long-read mapping methods often generate wrong alignments and variant calls. Compared to deletions and insertions, inversion events are more difficult to be detected since the anchors in inversion regions are nonlinear to those in SV-free regions. To address this issue, this study presents a novel long-read mapping algorithm (named as invMap). RESULTS: For each long noisy read, invMap first locates the aligned region with a specifically designed scoring method for chaining, then checks the remaining anchors in the aligned region to discover potential inversions. We benchmark invMap on simulated datasets across different genomes and sequencing coverages, experimental results demonstrate that invMap is more accurate to locate aligned regions and call SVs for inversions than the competing methods. The real human genome sequencing dataset of NA12878 illustrates that invMap can effectively find more candidate variant calls for inversions than the competing methods. AVAILABILITY AND IMPLEMENTATION: The invMap software is available at https://github.com/zhang134/invMap.git.
Subject(s)
Genomics , High-Throughput Nucleotide Sequencing , Humans , Genomics/methods , High-Throughput Nucleotide Sequencing/methods , Software , Algorithms , Genome, Human , Chromosome Inversion , Sequence Analysis, DNA/methodsABSTRACT
Peripheral T-cell lymphoma is a disease that includes multiple T-cell lymphoma subtypes. It is still unclear whether CD30 can be used as a new target molecule and classification standard for PTCL. Differences in the molecular characteristics of CD30-positive PTCL and CD30-negative PTCL have rarely been reported. This study aimed to analyze the expression of BCL11b and CDKN2A in CD30-positive PTCL and CD30-negative PTCL, in order to guide the pathological classification, prognosis, and clinical treatment of PTCL. Immunohistochemical staining and quantitative reverse-transcription PCR (qRT-PCR) were performed on formalin-fixed paraffin-embedded tissue. Verification of BCL11b and CDKN2A expression in ALCL, PTCL-NOS, AITL and NK/TCL. Based on immunohistochemical analysis, the expression level of BCL11b in the lymph node reactive hyperplasia control group was high at 85.0%, which was higher than 68.8% in CD30-positive PTCL and 44.1% in CD30-negative PTCL (Pâ <â .05, respectively). CDKN2A showed expression rates of 70.0% in the control group, 79.2% in CD30-positive PTCL and 79.4% in CD30-negative PTCL. qRT-PCR showed that the relative BCL11b mRNA expression levels in patients with PTCL were lower than those in the control group (0.694 vs 1.832, Pâ =â .045). Univariate analysis showed that international prognostic index score, CD30 expression, and BCL11b expression were closely related to prognosis (Pâ <â .05, respectively). Multivariate Cox regression analysis revealed that high expression of BCL11b mRNA was an independent factor affecting prognosis (respectively, Pâ <â .05). Spearman correlation analysis indicated that BCL11b expression had a significant positive correlation with CD30 expression (Pâ =â .005). These results indicate that BCL11b may be involved in CD30 differentiation and PTCL prognosis. The detection and targeting of BCL11b and CD30 may provide new strategies for the treatment and classification of PTCL.
Subject(s)
Lymphoma, T-Cell, Peripheral , Humans , Cyclin-Dependent Kinase Inhibitor p16/genetics , Ki-1 Antigen , Lymphoma, T-Cell, Peripheral/drug therapy , Prognosis , Retrospective Studies , RNA, Messenger , Transcription Factors , Tumor Suppressor ProteinsABSTRACT
PURPOSE: Programmed death-ligand 1 (PD-L1) and DNA mismatch repair (MMR) are considered predictive biomarkers for immunotherapy in cervical cancer. However, their expression in primary tumors and metastases does not always match affecting the course of treatment. We investigated the consistency of their expression in primary and matched recurrent/metastatic lesions from patients with cervical cancer. METHODS: Primary and matched recurrent/metastatic specimens from patients with recurrent cervical cancer (n = 194) were stained for PD-L1 and MMR (MLHI, MSH6, MSH2, and PMS2) using immunohistochemistry. The degree of consistency of PD-L1 and MMR expression in these lesions was analyzed. RESULTS: The inconsistency rate of PD-L1 expression in primary and recurrent/metastatic lesions was 33.0%, and it varied between the recurrence sites. Positive PD-L1 rate in primary lesions was lower (15.4%) than that in recurrent/metastatic lesions (30.4%). The discordance rate of MMR expression between primary and recurrent/metastatic lesions was 4.1%. CONCLUSION: We conclude that to use PD-L1 as a predictive biomarker for immunotherapy, analysis of both metastatic and primary lesions may be required. High consistency rate of MMR expression between primary and metastatic lesions suggests that testing primary lesions alone can be sufficient for guiding the course of therapy, thereby solving the difficulty of obtaining recurrent/metastatic specimens in clinic.
Subject(s)
B7-H1 Antigen , Uterine Cervical Neoplasms , Female , Humans , B7-H1 Antigen/metabolism , Biomarkers, Tumor/genetics , Uterine Cervical Neoplasms/genetics , DNA Mismatch Repair , Neoplasm Recurrence, Local/geneticsABSTRACT
BACKGROUND: Unlike other subtypes of breast cancer, triple negative breast cancer (TNBC) exhibits aggressive and metastatic behaviors and a lack of effective targeted therapeutics. (R)-9bMS, a small-molecule inhibitor of the non-receptor tyrosine kinase 2 (TNK2), significantly inhibited TNBC cell growth; however, the functional mechanism of (R)-9bMS in TNBC remains largely unknown. OBJECTIVE: To explore the functional mechanism of (R)-9bMS in TNBC. METHODS: Cell proliferation, apoptosis and xenograft tumor growth assays were performed to evaluate the effects of (R)-9bMS on TNBC. The expression levels of miRNA and protein were detected by RTqPCR or western blot, respectively. Protein synthesis was determined by analyzing the polysome profile and 35S-met incorporation. RESULTS: (R)-9bMS attenuated TNBC cell proliferation, induced cell apoptosis, and inhibited xenograft tumor growth. Mechanism study indicated that (R)-9bMS upregulated the expression of miR-4660 in TNBC cells. The expression of miR-4660 is lower in TNBC samples than that of the non-cancerous tissues. miR-4660 overexpression inhibited TNBC cell proliferation by targeting the mammalian target of rapamycin (mTOR), which reduced mTOR abundance in TNBC cells. Consistent with the downregulation of mTOR, exposure of (R)-9bMS inhibited the phosphorylation of p70S6K and 4E-BP1, which consequently interrupted the total protein synthesis and autophagy of TNBC cells. CONCLUSION: These findings uncovered the novel working mechanism of (R)-9bMS in TNBC by attenuating mTOR signaling via up-regulating miR-4660. The potential clinical significance of (R)- 9bMS in TNBC treatment is interesting to explore.
Subject(s)
MicroRNAs , Triple Negative Breast Neoplasms , Humans , Triple Negative Breast Neoplasms/genetics , Triple Negative Breast Neoplasms/metabolism , Triple Negative Breast Neoplasms/pathology , Cell Line, Tumor , MicroRNAs/genetics , MicroRNAs/metabolism , TOR Serine-Threonine Kinases/metabolism , Cell Proliferation , Autophagy , Gene Expression Regulation, Neoplastic , Cell Movement , Protein-Tyrosine Kinases/genetics , Protein-Tyrosine Kinases/metabolism , Protein-Tyrosine Kinases/pharmacologyABSTRACT
BACKGROUND: Anaplastic carcinoma mural nodules in ovarian mucinous tumors are very rare. This study aimed to report the morphological characteristics, molecular detection results, clinical treatment and prognosis of three ovarian mucinous tumors with mural nodules of anaplastic carcinoma. CASE SUMMARY: The pathomorphological features, molecular detection results, clinical treatment and prognosis of anaplastic carcinoma mural nodules were described in three cases. In case 1, sarcoma-like mural nodules (SLMNs) coexisted with anaplastic carcinoma mural nodules. No mutation was found in mucinous tumors. KRAS mutation was found in anaplastic carcinoma nodules and heterotypic cells were found in SLMNs. In case 2, KRAS mutation occurred in the mucinous epithelium and BRAF mutation occurred in mural nodules. In case 3, both mural nodules and mucinous tumors had the same KRAS mutation and a morphological transition between them was observed. All three patients died within 2 years, whether receiving chemotherapy or not. CONCLUSION: Anaplastic carcinoma mural nodules may develop from dedifferentiation of mucinous tumors or are unrelated to mucinous tumors.
ABSTRACT
The molecular subtyping of triple-negative breast cancer (TNBC) is critical to guiding individualized patient treatment. In this study, we sought to characterize the clinicopathologic features of TNBC subtypes and to identify correlates of patient survival in an effort to provide a robust foundation for treatment planning. We additionally assessed PD-L1 expression in Chinese TNBC patients and evaluated the relationship between such expression and immunotherapeutic treatment outcomes. Based on analyses of histologic characteristics including apocrine differentiation, tumor-infiltrating lymphocytes, and metaplastic features, we selected immunohistochemical (IHC) markers including CD8, FOXC1, and AR for use in classifying TNBC cases. Associations between these subtypes and a range of clinicopathologic characteristics were evaluated. We classified a cohort of 93 TNBC patients into individuals with luminal androgen receptor (LAR), immunomodulatory (IM), basal-like immune-suppressed (BLIS), and mesenchymal (MES) tumor subtypes (23, 24, 39, and 7 cases, respectively). PD-L1 positivity was observed in 49.6% of cases and was more common in individuals with IM subtype disease. Mismatch repair deficiency (dMMR) was observed in just one patient. Significant differences in histologic grade, pT stage, lymphocyte distribution patterns, large scarring areas without cells in tumor of central (central scar), and PD-L1, P53, and Rb status were observed among these TNBC subtypes, whereas no such differences were observed with respect to age, invasion pattern, or pN stage. Rates of disease progression were higher at the 40-50 month follow-up time point, but there were no significant differences in recurrence-free survival or breast cancer-specific survival among these subtypes. IHC markers associated with clinicopathologic characteristics represent a powerful approach to TNBC molecular typing, providing a foundation for precision patient treatment. PD-L1 expression may represent a relevant factor in TNBC patient immunotherapeutic treatment planning, whereas dMMR is not likely to be of substantial value when evaluating immunotherapeutic efficacy in these patients.
Subject(s)
Triple Negative Breast Neoplasms , B7-H1 Antigen , Biomarkers, Tumor , Cohort Studies , Humans , Lymphocytes, Tumor-Infiltrating/pathology , Triple Negative Breast Neoplasms/metabolism , Triple Negative Breast Neoplasms/therapyABSTRACT
BACKGROUND: Secretory breast carcinoma is an uncommon subset of breast cancer that usually has a favorable outcome. Although initially described in children, it also occurs in adults where it may metastasize, possibly resulting in death. To date, only 20 cases of secretory breast carcinoma with distant metastases have been described. CASE PRESENTATION: A 42-year-old female presented with liver metastasis after modified radical mastectomy of the left breast in 2008 at 34 years of age. The liver metastasis was morphologically similar to the primary tumor. Pan-TRK and Fluorescence in situ hybridization showed a rearrangement in the ETV6 gene. She subsequently underwent adjuvant chemotherapy with a fatal outcome. CONCLUSIONS: Although secretory breast carcinoma is usually associated with favorable outcomes, our study and reviews provide a novel insight into the genetic spectrum and treatment of secretory breast carcinoma showing reduced expression of hormone receptors, abnormal genomic profiles, and possible poor prognosis. Targeted therapy may curb clinically aggressive cases. Additional molecular investigations are needed to determine the links between specific mutations and poor prognosis.
Subject(s)
Breast Neoplasms/pathology , Carcinoma/secondary , Liver Neoplasms/secondary , Adult , Biomarkers, Tumor/analysis , Biomarkers, Tumor/genetics , Breast Neoplasms/chemistry , Breast Neoplasms/genetics , Breast Neoplasms/secondary , Breast Neoplasms/therapy , Carcinoma/chemistry , Carcinoma/genetics , Carcinoma/therapy , Chemoradiotherapy, Adjuvant , Fatal Outcome , Female , Gene Rearrangement , Humans , Immunohistochemistry , In Situ Hybridization, Fluorescence , Liver Neoplasms/chemistry , Liver Neoplasms/genetics , Mastectomy, Modified Radical , Proto-Oncogene Proteins c-ets/genetics , Receptors, Nerve Growth Factor/analysis , Repressor Proteins/genetics , Treatment Outcome , ETS Translocation Variant 6 ProteinABSTRACT
PURPOSE: Mutations of PIK3CA have recently been shown to play an important role in the pathogenesis and progression of breast neoplasms. The prevalence of PIK3CA in Chinese breast cancer patients may be underestimated. Therefore, we investigated the distribution of somatic PIK3CA mutation in Chinese breast cancer patients and explored their role in tumor phenotypes. METHODS: Mutational analysis of PIK3CA was done in 113 primary breast cancers of Chinese women used Amplification refractory mutation system (ARMS). The relationship of PIK3CA mutations with several clinicopathologic characteristics was analyzed. RESULTS: PIK3CA gene mutation was identified in 43(38.05%) cases and has a more significant difference between exon 9 and 20. HER2 gene amplification was 32.6% in 43 cases of PIK3CA mutation, but 37.1% in 70 cases of non-mutation (χ2 = 0.245, P > 0.05). There was no significant correlation of the age distribution, lymph node status, histological tumor grading, ER and/or PR and P53 between 2 groups (P > 0.05). CONCLUSION: A high frequency of somatic PIK3CA mutation was detected in Chinese breast cancer patients, especially in exon 20. The relationship between PIK3CA gene mutation and clinical pathological features of breast cancer needs to be further studied in a large series of patients. PIK3CA mutations seem to have the potential to be used in target treatment and as an indicator of prognosis.
Subject(s)
Breast Neoplasms/genetics , Breast Neoplasms/pathology , Class I Phosphatidylinositol 3-Kinases/genetics , Mutation , Adult , Aged , Aged, 80 and over , China , Class I Phosphatidylinositol 3-Kinases/metabolism , DNA Mutational Analysis , Female , Gene Amplification , Humans , Immunohistochemistry , In Situ Hybridization, Fluorescence , Middle Aged , Mutation Rate , Neoplasm Grading , Neoplasm Staging , PrognosisABSTRACT
The tumor microenvironment contributes to the survival and development of tumor cells and is therefore a key target for cancer therapy. The tumor microenvironment has unique physical and chemical properties and is associated with inflammation and immunity. To examine the correlation between tumor microenvironment-associated factors and the efficacy and prognosis of neoadjuvant therapy for rectal cancer, and to compare the differences between two treatments [neoadjuvant chemotherapy (NAC) vs. neoadjuvant chemoradiotherapy (NACR)], an immunohistochemical method was used to measure the expression levels of CD4+ tumor-infiltrating lymphocytes (TILs), cluster of differentiation (CD)8+TILs, forkhead box P3 (FOXP3)+TILs, cytotoxic T lymphocyte-associated antigen-4+TILs and programmed death ligand-1 (PD-L1)+TILs in 109 patients with rectal cancer, pre- and post-neoadjuvant therapy. The significance of these protein expression patterns was also analyzed using tissue microarrays, and the prognostic significance of these findings evaluated. The results indicated that high levels of CD4+TILs, CD8+TILs and PD-L1+TILs may be associated with favorable responses to neoadjuvant therapy, whereas high levels of FOXP3+TILs were associated with poor therapeutic responses. Expression levels of CD8+TILs and FOXP3+TILs following neoadjuvant therapy were independent prognostic factors and affected the total survival of patients subjected to neoadjuvant therapy for the treatment of rectal cancer. Moreover, the effects of NAC and NACR on the tumor microenvironment may be different.
ABSTRACT
BACKGROUND: Three-dimensional (3D) printing technology holds great promise for treating diseases or injuries that affect human bones with enhanced performance over traditional techniques. Different patterns of design can lead to various mechanical properties and biocompatibility to various degrees. However, there is still a long way to go before we can fully take advantage of 3D printing technologies. METHODS: This study tailored 3D printed scaffolds with gelatin and platelets to maximize bone regeneration. The scaffolds were designed with special internal porous structures that can allow bone tissue and large molecules to infiltrate better into the scaffolds. They were then treated with gelatin and platelets via thermo-crosslinking and freeze-drying, respectively. Vascular endothelial growth factor (VEGF) and transforming growth factor (TGF)-ß1 were measured at different time points after the scaffolds had been made. Cell proliferation and cytotoxicity were determined via cell counting kit-8 (CCK-8) assay. RESULTS: There was a massive boost in the level of VEGF and TGF-ß1 released by the scaffolds with gelatin and platelets compared to that of scaffolds with only gelatin. After 21 days of culture, the CCK-8 cell counts of the control group and treated group were significantly higher than that of the blank group (P < 0.05). The cytotoxicity test also indicated the safety of the scaffolds. CONCLUSIONS: Our experiments confirmed that the 3D printed scaffolds we had designed could provide a sustained-release effect for growth factors and improve the proliferation of preosteoblasts with little cytotoxicity in vitro. They may hold promise as bone graft substitute materials in the future.
Subject(s)
Biocompatible Materials/chemistry , Gelatin/chemistry , Printing, Three-Dimensional , Tissue Engineering/methods , Tissue Scaffolds/chemistry , 3T3 Cells , Animals , Cell Proliferation/drug effects , Cell Survival/drug effects , Mice , Transforming Growth Factor beta1/chemistry , Transforming Growth Factor beta1/pharmacology , Vascular Endothelial Growth Factor A/chemistry , Vascular Endothelial Growth Factor A/pharmacologyABSTRACT
<p><b>BACKGROUND</b>Three-dimensional (3D) printing technology holds great promise for treating diseases or injuries that affect human bones with enhanced performance over traditional techniques. Different patterns of design can lead to various mechanical properties and biocompatibility to various degrees. However, there is still a long way to go before we can fully take advantage of 3D printing technologies.</p><p><b>METHODS</b>This study tailored 3D printed scaffolds with gelatin and platelets to maximize bone regeneration. The scaffolds were designed with special internal porous structures that can allow bone tissue and large molecules to infiltrate better into the scaffolds. They were then treated with gelatin and platelets via thermo-crosslinking and freeze-drying, respectively. Vascular endothelial growth factor (VEGF) and transforming growth factor (TGF)-β1 were measured at different time points after the scaffolds had been made. Cell proliferation and cytotoxicity were determined via cell counting kit-8 (CCK-8) assay.</p><p><b>RESULTS</b>There was a massive boost in the level of VEGF and TGF-β1 released by the scaffolds with gelatin and platelets compared to that of scaffolds with only gelatin. After 21 days of culture, the CCK-8 cell counts of the control group and treated group were significantly higher than that of the blank group (P < 0.05). The cytotoxicity test also indicated the safety of the scaffolds.</p><p><b>CONCLUSIONS</b>Our experiments confirmed that the 3D printed scaffolds we had designed could provide a sustained-release effect for growth factors and improve the proliferation of preosteoblasts with little cytotoxicity in vitro. They may hold promise as bone graft substitute materials in the future.</p>
Subject(s)
Animals , Mice , 3T3 Cells , Biocompatible Materials , Chemistry , Cell Proliferation , Cell Survival , Gelatin , Chemistry , Printing, Three-Dimensional , Tissue Engineering , Methods , Tissue Scaffolds , Chemistry , Transforming Growth Factor beta1 , Chemistry , Pharmacology , Vascular Endothelial Growth Factor A , Chemistry , PharmacologyABSTRACT
A novel full-range Fourier domain Doppler optical coherence tomography (full-range FD-DOCT) using sinusoidal phase modulation for B-M scan is proposed. In this sinusoidal B-M scan, zero optical path difference (OPD) position does not move corresponding to lateral scanning points in contrast to linear B-M scan. Since high phase sensitivity arises around the zero OPD position, the proposed full-range FD-DOCT can achieve easily high velocity sensitivity without mirror image around the zero OPD position. Velocity sensitivity dependent on the OPD and the interval of scanning points is examined, and flow velocity detection capability is verified through Doppler imaging of a flow phantom and an in vivo biological sample.
Subject(s)
Blood Flow Velocity/physiology , Ear/physiology , Image Interpretation, Computer-Assisted/instrumentation , Laser-Doppler Flowmetry/instrumentation , Tomography, Optical Coherence/instrumentation , Animals , Ear/blood supply , Equipment Design , Equipment Failure Analysis , Image Interpretation, Computer-Assisted/methods , Laser-Doppler Flowmetry/methods , Mice , Reproducibility of Results , Sensitivity and Specificity , Tomography, Optical Coherence/methodsABSTRACT
Eighteen lignans were isolated from an ethanol extract of Machilus robusta by a combination of various chromatographic techniques including column chromatography over silica gel, Sephadex LH-20 and reversed-phase HPLC. Their structures were identified by spectroscopic data analysis as isolariciresinol-9'-O-beta-D-xylopyranoside (1), (+)-5'-methoxy-isolariciresinol-9'-O-beta-D-xylopyranoside(2), lyoniresinol-9'-O-beta-D-xylopyranoside(3), (+)-(8S, 8'S) -4, 4'-dihydroxy-3, 3', 5, 5'-tetramethoxylignan-9, 9'-diol 9-O-beta-D-xylopyranoside (ssioriside, 4), lyoniresinol (5), meso-dihydroguaiaretic acid (6), (+)-(8S, 8'R)-3', 4, 4'-trihydroxy-3'-methoxylignan (7), (8S, 8'R)-4'-hydroxy-3, 3', 4-trimethoxylignan (meso-monomethyl dihydroguaiaretic acid, 8), (+)-guaiacin (9), isoguaiacin (10), (-)-(7'R, 8R, 8'R)-4, 4'-dihydroxy-3, 3', 5-trimethoxy-2, 7'-cyclolignan (11), henricine B (12), (-)-(7S, 7'S, 8R, 8'R)-4, 4'-dihydroxy-3, 3', 5, 5'-tetramethoxy-7, 7'-epoxylignan-9, 9'-dio] (7S, 7'S, 8R, 8'R-icariol A2, 13), (+)-(7R, 8R, 7'E)-4-hydroxy-3, 5'-dimethoxy-7, 4'-epoxy-8, 3'-neolignan-7'-ene (licarin A, 14), nectandrin B (15), machilin-I (16), (-)-pinoresinol (17), and (-)-syringaresinol (18). All compounds were isolated from this plant for the first time. In the preliminary assay, compound 17 showed inhibitory activity against NO secretion of mouse peritoneal macrophages with an inhibition rate of 72.2% at 10 micromol x L(-1).
Subject(s)
Lauraceae/chemistry , Lignans/chemistry , Plant Extracts/chemistry , Mass Spectrometry , Molecular StructureABSTRACT
Thirty-three compounds were isolated from the root decoction of Isatis indigotica by using a combination of various chromatographic techniques including silica gel, macroporous adsorbent resin, Sephadex LH-20, and reversed-phase HPLC. Their structures were elucidated by spectroscopic data as (+)-dehydrovomifoliol (1), (S)-(+)-abscisic acid (2), vomifoliol (3), cyclo (L-Phe-L-Leu) (4), cyclo(L-Phe-L-Tyr) (5), cyclo(L-Tyr-L-Leu) (6), cyclo(L-Pro-L-Tyr) (7), evofolin B (8), (+)-syringaresinol (9), (-)-(7R,7'R,8S,8'S)-4,4'-dihydroxy-3-methoxy-7,9';7',9-diepoxy-lignan (10), (-)-medioresinol (11), (+) -(7R,7'R,8S,8'S) -neo-olivil (12), (-) -5-methoxyisolariciresinol (13), 1,3-dihydro-2H-indol-2-one (14), isalexin (15), dihydroneoascorbigen (16), indican (17), (-) -(S) -cyanomethyl-3-hydroxyoxindole (18), isoformononetein (19), calycosin (20), stigamast-5-ene-3beta-ol-7-one (21), acetovanillone (22), 3, 5-dimethoxy-4-hydroxyacetophenone (23), dihydroconiferyl alcohol (24), dihyroferulic acid (25), 3-hydroxy-1-(4-hydroxyphenyl) propan-1-one (26), beta-hydroxypropiovanillone (27), 4-aminobenzoic acid (28), 3-(4-hydroxyphenyl) propan-1-ol (29), 4-(2-hydroxyethyl) phenol (30), 2-methoxy-4-vinylphenol (31), pyrocatechol (32), and 4-pentenamide (33). These compounds were isolated from the root of I. indigotica for the first time. In preliminary in vitro assays, compound 19 showed activity against the influenza virus A/Hanfang/359/95 (H3N2), the herpes simplex virus 1 (HSV-1), and Coxsackie virus B3 (Cox-B3), with IC50 values of 2.06, 6.84, and 8.70 micromol x L(-1), respectively, but other compounds were in-active at a concentration of 1.0 x 10 x (-5) mol x L(-1).
Subject(s)
Isatis/chemistry , Plant Extracts/chemistry , Plant Roots/chemistry , Animals , Cell Line , Humans , Plant Extracts/pharmacology , Plant Extracts/toxicityABSTRACT
Six new glycosidic constituents (1-6), together with 10 known analogs, have been isolated from the bark of Machilus robusta. Structures of the new compounds, including the absolute configurations, were determined by spectroscopic and chemical methods as ( - )-nectandrin B-ß-d-glucopyranoside (1), ( - )-(7R,7'R,8S,8'R)-4,4'-dihydroxy-3,3'-dimethoxy-7,7'-epoxylignan-4-O-ß-d-glucopyranoside (2), ( - )-(7R,7'R,8S,8'R)-4,4'-dihydroxy-3,3'-dimethoxy-7,7'-epoxylignan-4'-O-ß-d-glucopyranoside (3), ( - )-(8S,8'R)-4,4'-dihydroxy-3,3',5'-trimethoxylignan-4'-O-ß-d-glucopyranoside (4), ( - )-(7R,8R)-syringylglycerol-8-O-ß-d-glucopyranoside (5), and ( - )-3-hydroxy-2-methyl-4-pyrone-3-O-ß-d-xylopyranosyl-(1 â 6)-O-ß-d-glucopyranoside (6), respectively.
Subject(s)
Glycosides/isolation & purification , Lauraceae/chemistry , Drugs, Chinese Herbal/chemistry , Glycosides/chemistry , Molecular Structure , Plant Bark/chemistry , StereoisomerismABSTRACT
Six new glycosides (1-6) have been isolated from the flower buds of Lonicera japonica. Their structures including the absolute configurations were determined by spectroscopic and chemical methods as ( - )-2-hydroxy-5-methoxybenzoic acid 2-O-ß-d-(6-O-benzoyl)-glucopyranoside (1), ( - )-4-hydroxy-3,5-dimethoxybenzoic acid 4-O-ß-d-(6-O-benzoyl)-glucopyranoside (2), ( - )-(E)-3,5-dimethoxyphenylpropenoic acid 4-O-ß-d-(6-O-benzoyl)-glucopyranoside (3), ( - )-(7S,8R)-(4-hydroxyphenylglycerol 9-O-ß-d-[6-O-(E)-4-hydroxy-3,5-dimethoxyphenylpropenoyl]-glucopyranoside (4), ( - )-(7S,8R)-(4-hydroxy-3-methoxyphenylglycerol 9-O-ß-d-[6-O-(E)-4-hydroxy-3,5-dimethoxyphenylpropenoyl]-glucopyranoside (5), and ( - )-4-hydroxy-3-methoxyphenol ß-d-{6-O-[4-O-(7S,8R)-(4-hydroxy-3-methoxyphenylglycerol-8-yl)-3-methoxybenzoyl]}-glucopyranoside (6), respectively.