A multiplexed in vivo approach to identify driver genes in small cell lung cancer.

Small cell lung cancer (SCLC) is a lethal form of lung cancer. Here, we develop a quantitative multiplexed approach on the basis of lentiviral barcoding with somatic CRISPR-Cas9-mediated genome editing to functionally investigate candidate regulators of tumor initiation and growth in genetically engineered mouse models of SCLC. We found that naphthalene pre-treatment enhances lentiviral vector-mediated SCLC initiation, enabling high multiplicity of tumor clones for analysis through high-throughput sequencing methods. Candidate drivers of SCLC identified from a meta-analysis across multiple human SCLC genomic datasets were tested using this approach, which defines both positive and detrimental impacts of inactivating 40 genes across candidate pathways on SCLC development. This analysis and subsequent validation in human SCLC cells establish TSC1 in the PI3K-AKT-mTOR pathway as a robust tumor suppressor in SCLC. This approach should illuminate drivers of SCLC, facilitate the development of precision therapies for defined SCLC genotypes, and identify therapeutic targets.

The association between serum vitamin A and NAFLD among US adults varied in different BMI groups: a cross-sectional study.

Background and aims: The association between serum vitamin A (VA) and non-alcoholic fatty liver disease (NAFLD) has not been adequately studied. This study aimed to evaluate the association between them in different BMI groups among US adults. Methods: A cross-sectional study was performed using the 2017-2018 National Health and Nutrition Examination Survey (NHANES) datasets (N = 4.723). Linear/logistic regression, interaction effect and mediation analyses were adopted to analyze the association. Results: NAFLD tended to be more prevalent in adults in the middle and high tertiles of serum VA than in those in the low tertile of serum VA (OR [95% CI], 1.17 [0.94, 1.45] and 1.43 [1.16, 1.75]). In the sensitivity analysis, subjects in the middle or high tertile of serum VA had 10% (OR, 1.10 [0.88, 1.39] and 31% (OR, 1.31 [1.09, 1.58]) higher odds of NAFLD than those in the low tertile of serum VA. In the normal weight group, higher serum VA was associated with 125% and 333% higher odds of NAFLD in the middle and high tertiles, respectively, (OR, 2.25 [1.46, 3.48] and 4.33 [2.43, 7.69]) compared with the low tertile serum VA group. However, serum VA and NAFLD were not significantly associated with the obese group. Among different BMI groups (<30 compared with ≥30), serum triglycerides and insulin resistance mediated the association between VA and NAFLD in adults to varying degrees. Conclusions: In the weighted survey, serum VA was positively associated with the degree of NAFLD, especially in the non-obese population.

Molecular classification and biomarkers of clinical outcome in breast ductal carcinoma in situ: Analysis of TBCRC 038 and RAHBT cohorts.

Ductal carcinoma in situ (DCIS) is the most common precursor of invasive breast cancer (IBC), with variable propensity for progression. We perform multiscale, integrated molecular profiling of DCIS with clinical outcomes by analyzing 774 DCIS samples from 542 patients with 7.3 years median follow-up from the Translational Breast Cancer Research Consortium 038 study and the Resource of Archival Breast Tissue cohorts. We identify 812 genes associated with ipsilateral recurrence within 5 years from treatment and develop a classifier that predicts DCIS or IBC recurrence in both cohorts. Pathways associated with recurrence include proliferation, immune response, and metabolism. Distinct stromal expression patterns and immune cell compositions are identified. Our multiscale approach employed in situ methods to generate a spatially resolved atlas of breast precancers, where complementary modalities can be directly compared and correlated with conventional pathology findings, disease states, and clinical outcome.

Trends in Iodine Status Among U.S. Children and Adults: A Cross-Sectional Analysis of National Health and Nutrition Examination Survey Data from 2001-2004 to 2017-2020.

Background: Iodine nutrition is an important public health issue. Trends in iodine status over time among U.S. schoolchildren and adults and factors mediating changes of iodine status were examined. Methods: In this cross-sectional study of National Health and Nutrition Examination Survey (NHANES) data, we estimated trends in the U.S. population using linear regression analyses. Representative samples of U.S. children and adults were enrolled in NHANES 2001-2020. The NHANES cycles were categorized into 5 four-year periods: 2001-2004, 2005-2008, 2009-2012, 2013-2016, and 2017-2020. The final sample sizes of children and adults for analysis were 4288 and 19,661, respectively. The estimated average requirement (EAR) (based on guidelines from the Institute of Medicine), was used to estimate the prevalence rate of inadequate iodine intake. Binary logistic regression analyses were used to investigate the association between iodine status and contributing factors. Results: From 2001-2004 to 2017-2020, among children, urinary iodine concentration (UIC) decreased from 243 to 166 µg/L (ptrend = 0.0057) and prevalence of iodine intake below the EAR rose from 15.4% to 27.6%. In adults, the UIC decreased from 153 to 116 µg/L (ptrend < 0.001) and prevalence of iodine intake below the EAR rose from 15.0% to 17.9%. A higher prevalence rate of iodine intake below the EAR was observed in females compared with males (children, 24.0% vs. 16.5%, p < 0.001; adults, 20.0% vs. 11.1%, p < 0.001). Inadequate iodine intake was less frequent among non-Hispanic White and Hispanic compared with non-Hispanic Black in children and adults. Adults without thyroid problems had a higher prevalence of inadequate iodine intake than those with thyroid problems (16.0% vs. 13.0%, p = 0.001). Inadequate iodine intake was less likely in the children who "sometimes" and "often" consumed milk products compared with children who "never or rarely" consumed milk products (OR = 0.60 [CI 0.30-1.21] and OR = 0.24 [CI 0.13-0.43], respectively). The prevalence of inadequate iodine intake among adults reporting "sometimes" (OR = 0.70 [CI 0.58-0.83]) and "often" consuming milk products was lower than those who "never or rarely" consumed them (OR = 0.36 [CI 0.30-0.44]). Conclusions: In this weighted survey, the prevalence of inadequate iodine intake increased from 2001-2004 to 2017-2020 among U.S. school-age children and adults. Sex, race, thyroid problems, and a decreased intake of milk products were significantly associated with iodine intake below the EAR.

The microdissected gene expression landscape of nasopharyngeal cancer reveals vulnerabilities in FGF and noncanonical NF-κB signaling.

Nasopharyngeal cancer (NPC) is an Epstein-Barr virus (EBV)-positive epithelial malignancy with an extensive inflammatory infiltrate. Traditional RNA-sequencing techniques uncovered only microenvironment signatures, while the gene expression of the tumor epithelial compartment has remained a mystery. Here, we use Smart-3SEQ to prepare transcriptome-wide gene expression profiles from microdissected NPC tumors, dysplasia, and normal controls. We describe changes in biological pathways across the normal to tumor spectrum and show that fibroblast growth factor (FGF) ligands are overexpressed in NPC tumors, while negative regulators of FGF signaling, including SPRY1, SPRY2, and LGALS3, are down-regulated early in carcinogenesis. Within the NF-κB signaling pathway, the critical noncanonical transcription factors, RELB and NFKB2, are enriched in the majority of NPC tumors. We confirm the responsiveness of EBV-positive NPC cell lines to targeted inhibition of these pathways, reflecting the heterogeneity in NPC patient tumors. Our data comprehensively describe the gene expression landscape of NPC and unravel the mysteries of receptor tyrosine kinase and NF-κB pathways in NPC.

Bidirectional Mendelian Randomization Analysis for Vitamin D and Thyroid Peroxidase Antibody.

Purpose: Whether Vitamin D deficiency or insufficiency is associated with thyroid autoimmunity was debated for long time. This study was still to explore the causal relationship of 25 (OH) D with a thyroid peroxidase antibody (TPOAb). Methods: The data were obtained from a cross-sectional study, SPECT-China study, which was performed in 23 sites in East China during 2014 to 2016. 10636 participants were finally included in this study. Genotyped four 25 (OH) D-related and four TPOAb-associated single nucleotide polymorphisms (SNPs) created their genetic risk scores (GRS). Bidirectional mendelian randomization (MR) analysis was used in this study. Results: 25 (OH) D GRS was significantly associated with 25 (OH) D (B -0.093, 95% CI -0.111, -0.074) and TPOAb level (B 0.067, 95% CI 0.002 to 0.132). TPOAb GRS was significantly associated with TPOAb concentration (B 0.345, 95% CI 0.135 to 0.556), but not 25 (OH) D (B -0.030, 95% CI -0.091 to 0.030). Using 25 (OH) D_GRS as instrumental variable in the MR analysis, a causal relationship of genetically determined 25 (OH) D with increased TPOAb concentration (B -0.720, 95% CI -1.429 to -0.012). No relation was found between genetically instrumented TPOAb and 25 (OH) D. Conclusion: A higher VD_GRS was associated with higher risk of increased TPOAb concentration, which supports a causal association between decreased vitamin D and increased concentration of TPOAb in an eastern Chinese population.

Are ethnic differences, urinary iodine status, lead and cadmium exposure associated with thyroid autoimmunity and hypothyroid status? A cross-sectional study.

OBJECTIVE: We aimed to evaluate the effects of different ethnicities and potential environmental exposure on the prevalence of thyroid autoimmune status and hypothyroid status. DESIGN: The data were obtained from two cross-sectional studies. PARTICIPANTS: 2105 participants in Shanghai (Han) and 772 participants in Yunnan Honghe Prefecture (Han, Yi, Miao and Hani), aged 18-75 were enrolled. METHODS: Participants underwent several checkups, including urinary iodine concentration, blood lead (BPb) and blood cadmium (BCd), thyroid peroxidase antibody (TPOAb), thyroglobulin antibody (TgAb), thyroid stimulating hormone (TSH) as well as thyroid ultrasonography (US). Thyroid autoimmune status was defined as: antithyroid antibody positive (ATA+): TPOAb + or TgAb+; and ATA + and US+: TPOAb + or TgAb + together with characteristic US features. RESULTS: The standardised prevalence of thyroid autoimmune positivity in Yunnan were higher than those in Shanghai (TPOAb+: 13.56% vs 8.27%, p<0.001; TgAb+: 9.28% vs 7.09%, p=0.045; ATA+: 16.96% vs 11.10%, p<0.001; ATA + and US+: 8.96% vs 6.64%, p=0.036). For urinary iodine-to-creatinine ratio (UI/Cr), compared with the level of 100.00-199.99 µg/g, the level of ≥300.00 µg/g had a 1.5-fold risk for ATA + and US+ (OR 1.455, p=0.041). The levels of 200.00-299.99 µg/g and ≥300.00 µg/g were positively associated with hypothyroid status (OR 1.509, p=0.002 and OR 1.338, p=0.043). Compared with the first quartiles, the fourth quartiles of BPb were positively associated with TPOAb+: (OR 1.637, p=0.006), ATA+ (OR 1.435, p=0.025), ATA + and US+ (OR 1.641, p=0.013), hypothyroid status (OR 1.467, p=0.013) and TSH levels (B 0.092, p=0.021). The fourth quartile of BCd was positively associated with the prevalence of ATA+ (OR 1.427, p=0.036). CONCLUSIONS: Higher levels of UI/Cr, BPb and BCd may be associated with thyroid autoimmunity and hypothyroid status.

Transition to invasive breast cancer is associated with progressive changes in the structure and composition of tumor stroma.

Ductal carcinoma in situ (DCIS) is a pre-invasive lesion that is thought to be a precursor to invasive breast cancer (IBC). To understand the changes in the tumor microenvironment (TME) accompanying transition to IBC, we used multiplexed ion beam imaging by time of flight (MIBI-TOF) and a 37-plex antibody staining panel to interrogate 79 clinically annotated surgical resections using machine learning tools for cell segmentation, pixel-based clustering, and object morphometrics. Comparison of normal breast with patient-matched DCIS and IBC revealed coordinated transitions between four TME states that were delineated based on the location and function of myoepithelium, fibroblasts, and immune cells. Surprisingly, myoepithelial disruption was more advanced in DCIS patients that did not develop IBC, suggesting this process could be protective against recurrence. Taken together, this HTAN Breast PreCancer Atlas study offers insight into drivers of IBC relapse and emphasizes the importance of the TME in regulating these processes.

Atlas of clinically distinct cell states and ecosystems across human solid tumors.

Determining how cells vary with their local signaling environment and organize into distinct cellular communities is critical for understanding processes as diverse as development, aging, and cancer. Here we introduce EcoTyper, a machine learning framework for large-scale identification and validation of cell states and multicellular communities from bulk, single-cell, and spatially resolved gene expression data. When applied to 12 major cell lineages across 16 types of human carcinoma, EcoTyper identified 69 transcriptionally defined cell states. Most states were specific to neoplastic tissue, ubiquitous across tumor types, and significantly prognostic. By analyzing cell-state co-occurrence patterns, we discovered ten clinically distinct multicellular communities with unexpectedly strong conservation, including three with myeloid and stromal elements linked to adverse survival, one enriched in normal tissue, and two associated with early cancer development. This study elucidates fundamental units of cellular organization in human carcinoma and provides a framework for large-scale profiling of cellular ecosystems in any tissue.

Transcriptome and genome evolution during HER2-amplified breast neoplasia.

BACKGROUND: The acquisition of oncogenic drivers is a critical feature of cancer progression. For some carcinomas, it is clear that certain genetic drivers occur early in neoplasia and others late. Why these drivers are selected and how these changes alter the neoplasia's fitness is less understood. METHODS: Here we use spatially oriented genomic approaches to identify transcriptomic and genetic changes at the single-duct level within precursor neoplasia associated with invasive breast cancer. We study HER2 amplification in ductal carcinoma in situ (DCIS) as an event that can be both quantified and spatially located via fluorescence in situ hybridization (FISH) and immunohistochemistry on fixed paraffin-embedded tissue. RESULTS: By combining the HER2-FISH with the laser capture microdissection (LCM) Smart-3SEQ method, we found that HER2 amplification in DCIS alters the transcriptomic profiles and increases diversity of copy number variations (CNVs). Particularly, interferon signaling pathway is activated by HER2 amplification in DCIS, which may provide a prolonged interferon signaling activation in HER2-positive breast cancer. Multiple subclones of HER2-amplified DCIS with distinct CNV profiles are observed, suggesting that multiple events occurred for the acquisition of HER2 amplification. Notably, DCIS acquires key transcriptomic changes and CNV events prior to HER2 amplification, suggesting that pre-amplified DCIS may create a cellular state primed to gain HER2 amplification for growth advantage. CONCLUSION: By using genomic methods that are spatially oriented, this study identifies several features that appear to generate insights into neoplastic progression in precancer lesions at a single-duct level.

Influence of Rapid Urbanization on Thyroid Autoimmune Disease in China.

BACKGROUND: The prevalence of autoimmune thyroid diseases (AITDs), especially Hashimoto's thyroiditis (HT), has increased dramatically in China. Moreover, China is experiencing the largest scale of urbanization in the world. We intended to explore the relationship between rapid urbanization and HT. METHODS: A total of 2946 subjects in Zhejiang Shangyu (SY) (n = 1546) and Jiangsu Nanjing (NJ) (n = 1400) were enrolled in this study. Serum TPOAb, TGAb, and thyrotropin (TSH) were measured, and ultrasonography of the thyroid was performed in all subjects. DNA was extracted from all subjects, and four SNPs were selected for genotyping. Generalized multifactor dimensionality reduction (GMDR) was used to screen the best interaction between genetic factors and environment factors. RESULTS: TPOAb and TGAb concentrations were higher in NJ than in SY (34.60 vs. 14.00 IU/ml and 21.05 vs. 7.50 IU/ml). People in NJ also had higher TPOAb and TGAb positivity rates than those in SY (7.8% vs. 12.7% and 8.7% vs. 16.3%). Logistic regression analysis indicated that rapid urbanization was an independent risk factor for TPOAb (OR = 1.473) and TGAb (OR = 1.689). Genotype TT in rs11675434 was associated with an increased risk of TPOAb positivity both in SY (OR = 2.955) and in NJ (OR = 1.819). GMDR analysis showed a two-locus model (SNP2 × urbanization) and a three-locus model (SNP2 × SNP3 × urbanization), which had testing accuracies of 56.88% and 57.25%, respectively (P values were 0.001 and 0.001). CONCLUSION: Rapid urbanization influences the incidence of TPOAb and TGAb positivity. We should pay more attention to thyroid autoimmune disease in areas of China experiencing rapid urbanization.

Association of Insulin Resistance and ß-cell Function With Bone Turnover Biomarkers in Dysglycemia Patients.

Background: The interrelation between glucose and bone metabolism is complex and has not been fully revealed. This study aimed to investigate the association between insulin resistance, ß-cell function and bone turnover biomarker levels among participants with abnormal glycometabolism. Methods: A total of 5277 subjects were involved through a cross-sectional study (METAL study, http://www.chictr.org.cn, ChiCTR1800017573) in Shanghai, China. Homeostasis model assessment of insulin resistance (HOMA-IR) and ß-cell dysfunction (HOMA-%ß) were applied to elucidate the nexus between ß-C-terminal telopeptide (ß-CTX), intact N-terminal propeptide of type I collagen (P1NP) and osteocalcin (OC). ß-CTX, OC and P1NP were detected by chemiluminescence. Results: HOMA-IR was negatively associated with ß-CTX, P1NP and OC (regression coefficient (ß) -0.044 (-0.053, -0.035), Q4vsQ1; ß -7.340 (-9.130, -5.550), Q4vsQ1 and ß -2.885 (-3.357, -2.412), Q4vsQ1, respectively, all P for trend <0.001). HOMA-%ß was positively associated with ß-CTX, P1NP and OC (ß 0.022 (0.014, 0.031), Q4vsQ1; ß 6.951 (5.300, 8.602), Q4vsQ1 and ß 1.361 (0.921, 1.800), Q4vsQ1, respectively, all P for trend <0.001). Conclusions: Our results support that lower bone turnover biomarker (ß-CTX, P1NP and OC) levels were associated with a combination of higher prevalence of insulin resistance and worse ß-cell function among dysglycemia patients. It is feasible to detect bone turnover in diabetes or hyperglycemia patients to predict the risk of osteoporosis and fracture, relieve patients' pain and reduce the expenses of long-term cure.

Relationship between Gene Polymorphisms and Urine Iodine Levels on Susceptibility to Thyroid Peroxidase Antibody Positivity in the Chinese Population.

BACKGROUND/AIMS: Hashimoto thyroiditis, characterized by positive thyroid peroxidase antibodies (TPOAbs), is caused by the interaction of genetic and environment factors. The aim of this study was to clarify the interaction of gene polymorphisms and iodine intake in the incidence of TPOAb positivity. METHODS: 1,733 subjects were included in this study. Genomic DNA was extracted from peripheral blood white cells. Four SNPs (rs11675434 [TPO], rs3094228 [HCP5], rs9277555 [HLA-DPB1], and rs301799 [RERE]) were selected for genotyping. Weighted TPOAb genetic risk score (GRS) was calculated based on these 4 SNPs. Thyroid hormones and autoimmune antibodies (TPOAb and thyroglobulin antibody) were determined using the electrochemiluminescence immunoassay method. RESULTS: The mean serum thyrotropin level in TPOAb-positive subjects was higher than in TPOAb-negative subjects (p < 0.01). Genotype GG of rs9277555 was associated with an increased risk of TPOAb positivity (OR = 1.64, 5-95% CI 1.09, 2.47, p = 0.02). Genotype TT of rs11675434 showed marginal increased risk of TPOAb positivity (OR = 1.57, 5-95% CI 1.01, 2.43, p = 0.048). Logistic regression analysis showed TPOAb-GRS and rs9277555 were associated with TPOAb positivity (OR = 5.09, 5-95% CI 1.30, 19.91, p = 0.02 and OR = 1.30, 5-95% CI 1.05, 1.61, p = 0.02). Subjects with a high TPOAb-GRS had a 52% increased risk of TPOAb positivity compared to subjects with a low TPOAb-GRS (OR 1.52, 5-95% CI 1.05, 2.21, p = 0.03). CONCLUSION: TPOAb-GRS was associated with an increased risk of TPOAb positivity in a Chinese Han population. This effect might be attribute to rs9277555.

Vitamin D is associated with blood lead exposure through bone turnover in type 2 diabetes patients.

BACKGROUND: Bone is thought to be the reservoir of the human lead burden, and vitamin D is associated with bone turnover. We aimed to explore whether exposure to lower 25-hydroxy vitamin D (25(OH)D) levels was associated with higher blood lead levels (BLLs) by increasing the bone turnover rate in individuals with type 2 diabetes. METHODS: A total of 4103 type 2 diabetic men and postmenopausal women in Shanghai, China, were enrolled in 2018. Their 25(OH)D, ß-C-terminal telopeptide (ß-CTX), N-MID osteocalcin and procollagen type 1 N-peptide (P1NP) levels were detected. Their BLLs were determined by atomic absorption spectrometry. Mediation analyses were performed to identify the possible role that bone turnover played in the underlying mechanisms. RESULTS: In both the men and postmenopausal women, all three bone turnover markers were inversely associated with 25(OH)D and positively associated with the BLL (all P < 0.01) after adjusting for age, current smoking habits, metabolic parameters, duration of diabetes, vitamin D intake, and use of anti-osteoporosis medication. In the mediation analyses, none of the direct associations between 25(OH)D and BLL was significant for the three bone turnover markers, but all three bone turnover markers were found to be significant mediators of the indirect associations between 25(OH)D and BLL. CONCLUSION: The association between vitamin D and BLL was fully mediated by bone turnover markers in type 2 diabetic patients (mediation effect). This finding suggested that vitamin D may protect against blood lead exposure from the bone reservoir by decreasing bone turnover in individuals with type 2 diabetes.

Prevalence of hyperuricaemia in an Eastern Chinese population: a cross-sectional study.

OBJECTIVES: In the past decade, China has been characterised by large-scale urbanisation as well as rapid economic growth. The aim of this study was to further investigate the prevalence of hyperuricaemia (HUA) in an Eastern Chinese population. DESIGN: Cross-sectional study. SETTING: Survey of Prevalence in East China of Metabolic Diseases and Risk Factors China study. PARTICIPANTS: In this study, 12 770 residents from 22 sites in Eastern China were recruited. Finally, 9225 subjects were included. MAIN OUTCOME MEASURES: The serum levels of uric acid (UA), fasting plasma glucose (FPG), glycated haemoglobin and other metabolic parameters were tested. Waist circumference, weight, height and blood pressure were also measured. Questionnaires regarding smoking, drinking, education were collected from the subjects. HUA was defined as serum UA >420 µmol/L for men and >360 µmol/L for women. RESULTS: The prevalence of HUA in this Eastern Chinese population was 11.3% (9.9, 12.7) overall, 20.7% (17.7, 23.7) in men and 5.6% (4.3, 6.7) in women. The prevalence of HUA in urban subjects was higher than that in rural subjects (12.9 vs 10.8%, p<0.01). The prevalence of HUA was negatively and positively associated with age in men and women, respectively. Residents with high body mass index levels had a higher prevalence of HUA. In the logistic regression analysis, male sex, urban residency, total cholesterol, triglyceride, overweight, obesity, systolic blood pressure and low economic status were independently correlated with HUA. CONCLUSIONS: The estimated prevalence of HUA in this Eastern Chinese population was 11.3% (9.9, 12.7) overall and 20.7% (17.7, 23.7) and 5.6% (4.3, 6.7) in men and women, respectively. HUA has gradually become an important public health issue in China. TRIAL REGISTRATION NUMBER: ChiCTR-ECS-14005052.

Preparation of UV-Curable Low Surface Energy Polyurethane Acrylate/Fluorinated Siloxane Resin Hybrid Coating with Enhanced Surface and Abrasion Resistance Properties.

Low surface energy coatings have gained considerable attention due to their superior surface hydrophobic properties. However, their abrasion resistance and sustainability of surface hydrophobicity are still not very satisfactory and need to be improved. In this work, a series of utraviolet (UV)-curable fluorosiloxane copolymers were synthesized and used as reactive additives to prepare polyurethane acrylate coatings with low surface energy. The effect of the addition of the fluorinated graft copolymers on the mechanical durability and surface hydrophobicity of the UV-cured hybrid films during the friction-annealing treatment cycles was investigated. The results show that introducing fluorosiloxane additives can greatly enhance surface hydrophobicity of the hybrid film. With addition of 2 wt.% fluorosiloxane copolymers, the water contact angle (WCA) value of the hybrid film was almost tripled compared to that of the pristine PU film, increasing from 58° to 144°. The hybrid film also showed enhanced abrasion resistance and could withstand up to about 60 times of friction under a pressure of 20 kPa. The microstructure formed in the annealed film was found to contribute much to achieve better surface hydrophobicity. The polyurethane acrylate/fluorinated siloxane resin hybrid film prepared in this study exhibits excellent potential for applications in the low surface energy field.

Photodynamic antitumor activity of Ru(ii) complexes of imidazo-phenanthroline conjugated hydroxybenzoic acid as tumor targeting photosensitizers.

Two novel Ru(ii) polypyridyl complexes bearing imidazo-phenanthroline conjugated hydroxybenzoic acid groups were designed to enhance the tumor targeting ability as photosensitizers for photodynamic therapy. [Ru(bpy)2(phcpip)] (ClO4)2 (Ru-1) and [Ru(bpy)2(ohcpip)] (ClO4)2 (Ru-2) (bpy = 2,2'-bipyridine; phcpip = 2-(3-carboxyl-4-hydroxyphenyl) imidazo [4,5-f]phenanthroline; ohcpip = 2-(2-hydroxyl-3-carboxyphenyl) imidazo [4,5-f] [1,10] phenanthroline) were synthesized and their photodynamic antitumor activities were investigated. Both complexes displayed high photocytotoxicity toward cancerous cell lines HepG2, A549, MCF-7, and MDA-MB-231, but low photocytotoxicity toward normal cell lines GES-1 and Huvec. They were mainly localized at the nucleus of HepG2 cells after 24 h incubation, arrested the cell cycle at the G2/M phase and induced cancer cell apoptosis through reactive oxygen species (ROS) mediated pathways. Tumor targeting of the complexes is attributed to stronger molecular binding to DNA.

Gene expression profiling of single cells from archival tissue with laser-capture microdissection and Smart-3SEQ.

RNA sequencing (RNA-seq) is a sensitive and accurate method for quantifying gene expression. Small samples or those whose RNA is degraded, such as formalin-fixed paraffin-embedded (FFPE) tissue, remain challenging to study with nonspecialized RNA-seq protocols. Here, we present a new method, Smart-3SEQ, that accurately quantifies transcript abundance even with small amounts of total RNA and effectively characterizes small samples extracted by laser-capture microdissection (LCM) from FFPE tissue. We also obtain distinct biological profiles from FFPE single cells, which have been impossible to study with previous RNA-seq protocols, and we use these data to identify possible new macrophage phenotypes associated with the tumor microenvironment. We propose Smart-3SEQ as a highly cost-effective method to enable large gene expression profiling experiments unconstrained by sample size and tissue availability. In particular, Smart-3SEQ's compatibility with FFPE tissue unlocks an enormous number of archived clinical samples; combined with LCM it allows unprecedented studies of small cell populations and single cells isolated by their in situ context.

Structure and Activity of the Camellia oleifera Sapogenin Derivatives on Growth and Biofilm Inhibition of Staphylococcus aureus and Escherichia coli.

Sapogenin is the main block of Camellia oleifera saponin, which was purified and structurally modified by the C28 acylation reaction to synthesize 19 new derivatives. The growth and biofilm inhibition of Staphylococcus aureus and Escherichia coli was measured to evaluate their antibacterial effects. A three-dimensional quantitative structure-activity relationship (3D-QSAR) assay indicated that the antibacterial activities were significantly enhanced after sapogenin was modified with an aromatic ring or heterocyclic ring and electron-withdrawing substituents at the meta or para position. Among them, the derivative of sapogenin with a 2-mercapto-4-methyl-5-thiazolyl acetyl group obviously destroyed bacterial biofilm and made bacteria lysis. 3D-QSAR provides practical information for the structural design of sapogenin derivatives with strong antibacterial activity, and the C. oleifera sapogenin derivative 28-O-(2-mercapto-4-methyl-5-thiazolyl)-3ß,16α,21ß,22α-O-tetrahydroxy-oleantel-2-ene-23-aldehyde (S-16) is an effective candidate as an antibacterial agent for the prevention of bacterial resistance against antibiotics.