ZFHX3 acts as a tumor suppressor in prostate cancer by targeting FTO-mediated m6A demethylation.

Zinc-finger homeobox 3 (ZFHX3, also known as ATBF1) suppresses prostatic tumorigenesis. ZFHX3 is frequently found to have numerous deletions in human prostate cancer (PCa). However, the underlying molecular function of ZFHX3 during prostatic tumorigenesis is not well understood. N6-methyladenosine (m6A) modification in RNA plays a critical role in the development of cancers; however, the relationship between ZFHX3 and m6A modification is largely unknown in PCa. In this study, we found that ZFHX3 knockdown decreased total m6A levels through enhancing the transcriptional activity of FTO in PCa cells. Importantly, FTO inhibition suppressed cell proliferation and rescued the promoting function of ZFHX3 knockdown on cell proliferation. In vivo, we verified that FTO was upregulated and ZFHX3 was decreased in PCa patients and that a high level of ZFHX3 is indispensable for low FTO expression and is correlated with better patient survival. Through transcriptome sequencing and MeRIP sequencing, we revealed that E2F2 and CDKN2C were the direct targets of FTO-mediated m6A modification and ZFXH3 was required for the regulation of FTO on E2F2 and CDKN2C expression. Unexpectedly, we uncovered that ZFHX3 expression was in return regulated by FTO in an m6A-dependent way. These findings establish a novel crosstalk mechanism between ZFHX3 and FTO in prostatic tumorigenesis.

Corrigendum: Expression profiling of ALOG family genes during inflorescence development and abiotic stress responses in rice (Oryza sativa L.).

[This corrects the article DOI: 10.3389/fgene.2024.1381690.].

Expression profiling of ALOG family genes during inflorescence development and abiotic stress responses in rice (Oryza sativa L.).

The ALOG (Arabidopsis LSH1 and Oryza G1) family proteins, namely, DUF640 domain-containing proteins, have been reported to function as transcription factors in various plants. However, the understanding of the response and function of ALOG family genes during reproductive development and under abiotic stress is still largely limited. In this study, we comprehensively analyzed the structural characteristics of ALOG family proteins and their expression profiles during inflorescence development and under abiotic stress in rice. The results showed that OsG1/OsG1L1/2/3/4/5/6/7/8/9 all had four conserved helical structures and an inserted Zinc-Ribbon (ZnR), the other four proteins OsG1L10/11/12/13 lacked complete Helix-1 and Helix-2. In the ALOG gene promoters, there were abundant cis-acting elements, including ABA, MeJA, and drought-responsive elements. Most ALOG genes show a decrease in expression levels within 24 h under ABA and drought treatments, while OsG1L2 expression levels show an upregulated trend under ABA and drought treatments. The expression analysis at different stages of inflorescence development indicated that OsG1L1/2/3/8/11 were mainly expressed in the P1 stage; in the P4 stage, OsG1/OsG1L4/5/9/12 had a higher expression level. These results lay a good foundation for further studying the expression of rice ALOG family genes under abiotic stresses, and provide important experimental support for their functional research.

Applying Rasch analysis in refinement and validation of interpersonal skills measure for gifted children.

Background: Interpersonal characteristics of gifted adolescents is important because of the potential influence on individuals' psychological health and future professional success. Understanding the interpersonal characteristics requires valid and reliable measures. This study attempted to explicitly describe the application of Rasch analysis in the validation and development process of an existing measure of interpersonal skills among gifted adolescent in Ohio. Methods: We extensively evaluated the psychometric properties of the 40-item scale measuring the interpersonal competence of adolescents among the gifted population in Ohio (N = 127) using Rasch analysis. Multiple aspects of reliability and validity were tested including dimensionality, rating scale functioning, and fit statistics. Results: The internal consistency reliability of the scale was confirmed with an adequate fit to the Rasch model. However, the scale demonstrates relatively poor performance in terms of unidimensionality with our sample. Also, the rating scale categories were confusing given that our sample could not effectively distinguish some adjacent categories. Corresponding refinements have been made and the revised scale formed a meaningful linear progression with improved performance on unidimensionality, rating scale functioning, and fit statistics. Conclusion: The study provided evidence that the construct of interpersonal skills is measurable. Based upon the original Interpersonal Competence Questionnaires consists of 40 items, the author extracted and piloted a refined measure consisting of 31 items that performed a meaningful, theoretically consistent linear progression measure that could be used to measure the level of interpersonal skills of gifted children.

The latest edition of WHO and ELN guidance and a new risk model for Chinese acute myeloid leukemia patients.

Objective: Diagnosis classification and risk stratification are crucial in the prognosis prediction and treatment selection of acute myeloid leukemia (AML). Here, we used a database of 536 AML patients to compare the 4th and 5th WHO classifications and the 2017 and 2022 versions of ELN guidance. Methods: AML patients were classified according to the 4th and 5th WHO classifications, as well as the 2017 and 2022 versions of the European LeukemiaNet (ELN) guidance. Kaplan-Meier curves with log-rank tests were used for survival analysis. Results: The biggest change was that 25 (5.2%), 8 (1.6%), and 1 (0.2%) patients in the AML, not otherwise specified (NOS) group according to the 4th WHO classification, were re-classified into the AML-MR (myelodysplasia-related), KMT2A rearrangement, and NUP98 rearrangement subgroups based on the 5th WHO classification. Referring to the ELN guidance, 16 patients in the favorable group, six patients in the adverse group, and 13 patients in the intermediate group based on the 2017 ELN guidance were re-classified to the intermediate and adverse groups based on the 2022 ELN guidance. Regrettably, the Kaplan-Meier curves showed that the survival of intermediate and adverse groups could not be distinguished well according to either the 2017 or 2022 ELN guidance. To this end, we constructed a risk model for Chinese AML patients, in which the clinical information (age and gender), gene mutations (NPM1, RUNX1, SH2B3, and TP53), and fusions (CBFB::MYH11 and RUNX1::RUNX1T1) were included, and our model could help divide the patients into favorable, intermediate, and adverse groups. Conclusion: These results affirmed the clinical value of both WHO and ELN, but a more suitable prognosis model should be established in Chinese cohorts, such as the models we proposed.

Approaches to Learning of Preschool Children in China: A Comparison between Only Children and Non-Only Children.

Preschool children are at the initial stage of individual development and at a critical period in cultivating their approaches to learning. In the context of China's frequently adjusted birth policies, further research is necessary on children's approaches to learning in families of different sizes. A questionnaire survey was administered to 5454 only child and 4632 non-only child parents from the east, middle, and west of China. The study found that children's approaches to learning generally developed well, but non-only children's approaches to learning was significantly lower than those of only children. There are four profiles of approaches to learning for both the only children and the non-only children. This study also found that gender, social skills, family income, and type of preschool exert significant influences on children's approaches to learning. Parents' educational background had a significant influence on only children's approaches to learning but had no significant influence on non-only children's approaches to learning. We provide practical implications for promoting children's approaches to learning in families of different sizes.

A novel 5-gene prognostic signature to improve risk stratification of cytogenetically normal acute myeloid leukemia.

PURPOSE: Prognostic prediction is a challenging task in cytogenetically normal acute myeloid leukemia (CN-AML) patients. In this study, we aimed at developing a novel prognostic signature to predict and stratify the survival of CN-AML patients. METHODS: Using a training dataset (GSE12417), 5-gene prognostic signature was established to predict survival of CN-AML patients. The prognostic performance of this prognostic signature was further validated in testing dataset (TCGA CN-AML cohort) and validation dataset (GSE6891 CN-AML cohort). RESULTS: In training, testing and validation datasets, the increased 5-gene risk score was significantly related with inferior overall survival (OS) of patients, and the area under the receiver operating characteristic curve (AUC) demonstrated that our prognostic signature had overall prediction accuracy. The excellent prognostic value of the 5-gene prognostic signature was also supported by the comparison with three previously proposed prognostic models. For the intermediate-risk CN-AML patients and the CN-AML patients with FLT3 or NPM1 mutation, our model could also well dichotomize them into two subgroups with distinct prognosis. Multivariate analysis demonstrated that 5-gene risk score was the only independent risk factor in TCGA CN-AML cohort. Nomogram including the 5-gene risk score performed well in predicting 1-year, 2-year and 3-year OS. CONCLUSION: In summary, our novel 5-gene prognostic signature facilitated the improvement in risk stratification of CN-AML patients.

Type of Family Support for Infant and Toddler Care That Relieves Parenting Stress: Does the Number of Children Matter?

The present study aimed to investigate the status and relationships between family support for infant and toddler care and parenting stress, and to explore differences related to the number of children in the families. We conducted a survey among 13,390 Chinese parents who were randomly sampled from six provinces of China. Descriptive analysis, multivariate analysis of variance, and regression analysis indicated that (1) current family support for infant and toddler care in China is insufficient; (2) most Chinese parents reported moderate parenting stress, with the highest scores given for parental distress, followed by difficult child, and parent-child dysfunctional interaction; (3) the larger the number of children in the family, the less the family support for infant and toddler care, the greater the parenting stress; (4) there was a difference between the effects of family support for infant and toddler care on relieving parenting stress among families with different numbers of children. These findings indicate that different types of family support for infant and toddler care should be provided for families with different numbers of children, to ease parenting stress and promote the implementation of the government's current fertility policy.

Unusual Experience in the COVID-19 Pandemic: Young Children's Readjustment from Home to Preschool following School Closures in Different Risk Regions.

Children's readjustment to preschool following long-term school closures during the COVID-19 pandemic merits special attention. This study examined children's preschool readjustment using a survey of 1008 teachers in a high-risk region and 1399 teachers in a fluctuating-risk region of China. Results found are as follows. (1) children's preschool readjustment was at a medium level after the long-term school closures. However, children's preschool readjustment scores in the fluctuating-risk region were significantly lower than those in the high-risk region. (2) Children in both regions were divided into four profiles based on their preschool readjustment: low-level, middle-level, upper-middle-level, and high-level groups. (3) Preschool transition practices and teachers' turnover intention are common factors relating to preschool readjustment in both regions. Teachers' professional development support impacted children's preschool readjustment only in the high-risk region. The findings inform the design of targeted interventions to help children readjust to preschool across different risk regions.

[Construction and application of lentivirus overexpression vector with two labeling genes fused with CopGFP and PuroR].

Objective: To construct the lentivirus overexpression vector with two label genes fused with CopGFP and PuroR and to detect the emission of green fluorescence as well as resistance to puromycin in liver cancer cells infected with lentivirus packaged with the above vector. Methods: Firstly, two fragments containing copGFP and PuroR coding sequences were amplified from pCDH-CMV-MCS-copGFP and pLKO.1 respectively; secondly, the two amplified regions were fused with each other by recombinant PCR; thirdly, the fusion DNA fragment was cut and inserted into pCDH-CMV-MCS-copGFP vector, which was linearized with the same restriction endonuclease as used to digest fusion DNA fragment: BamH Ⅰ and Sal Ⅰ. The fusion region in the constructed vector was confirmed by DNA sequencing. The checked vector was co-transfected with package assistant plasmids, namely PLP1, PLP2 and VSVG into in 293T cells and the culture supernatant was subjected to centrifuge and infect liver cancer MHCC97H cells, which were then used to detect their resistance to puromycin (infected cells were treated with 1 mg/ml puromycin for 7 days after infection) and to observe green fluorescence emission in microscope. To determine its efficiency in expressing foreign target protein, the Sp1 coding region was inserted into the MCS sites of the vector, and Sp1 mRNA and protein expression levels were compared with the vehicle vector by RT-qPCR and Western blot. Results: The lentivirus overexpression vector with two label genes fused with CopGFP and PuroR was successfully constructed, and the liver cancer cells infected with lentivirus packaged with the vector expressing two labeling genes fused with CopGFP and PuroRshowed both emission of green fluorescence and resistance to puromycin simultaneously, while cells containing with the vector inserted with Sp1 coding region improved Sp1 mRNA level with 3.3 fold and protein level with 2.2 fold higher in comparison with cells containing the vehicle vector (P＜0.01). Conclusion: The fused label genes consisting of copGFP and PuroR are correctly cloned into the lentivirus vector and confer cells with the ability to emission of green fluorescence and resistance to puromycin, besides, the vector may promote the expression of the target gene with long coding sequence.

Bacillus velezensis EEAM 10B Strengthens Nutrient Metabolic Process in Black Soldier Fly Larvae (Hermetia illucens) via Changing Gut Microbiome and Metabolic Pathways.

Insects are a potential alternative protein source to solve the food shortage crisis. Previous studies have illustrated that probiotics can improve the substrate conversion efficiency of insects and increase insect protein content. However, the effects of probiotics on insect physiology and nutrient metabolism are still not well understood. Here, the black soldier fly larvae (BSFL), Hermetia illucens (Diptera: Stratiomyidae), was used as a study subject to deeply investigate the specific interaction among a novel probiotic, Bacillus velezensis EEAM 10B (10B), intestinal microbiota, and the host. In this study, the effects of 10B on the survival and physiology of BSFL were first analyzed. It shows that 10B significantly elevated the substrate conversion rate, average dry weight, and protein content of BSFL by 5%, 0.13 g/pc, and 8%, respectively. Then, we assessed the effect of 10B on the microbial community composition in the gut and frass of BSFL using Illumina Miseq sequencing. It shows that 10B significantly altered the microbial composition of the gut, but not that of the frass. Pearson's correlation analysis further showed that the Bacillus, unclassified_of_Caloramatoraceae, and Gracilibacillus were positively correlated with the survival rate, crude protein content, and substrate conversion rate of BSFL. To further investigate the effect of 10B on host metabolism, metabolic analyses on germ-free BSFL, monobacterial intestinal BSFL, and natural BSFL were also performed. The results proved that 10B (i) played a vital role in the survival of BSFL; and (ii) regulated the amino acid synthetic and metabolic process of BSFL, thus leading to the rise of the protein content of BSFL. In addition, vitamin backfill assays verified that the BSFL survival rate was significantly improved by supplying the germ-free BSFL with riboflavin, which further suggests that 10B determines the survival of BSFL via delivering riboflavin. Overall, this study provides a reference for understanding the comprehensive contribution of a specific probiotic to its host.

Cinnamaldehyde prevents intergenerational effect of paternal depression in mice via regulating GR/miR-190b/BDNF pathway.

Paternal stress exposure-induced high corticosterone (CORT) levels may contribute to depression in offspring. Clinical studies disclose the association of depressive symptoms in fathers with their adolescent offspring. However, there is limited information regarding the intervention for intergenerational inheritance of depression. In this study we evaluated the intervention of cinnamaldehyde, a major constituent of Chinese herb cinnamon bark, for intergenerational inheritance of depression in CORT- and CMS-induced mouse models of depression. Depressive-like behaviors were induced in male mice by injection of CORT (20 mg·kg-1·d-1, sc) for 6 weeks or by chronic mild stress (CMS) for 6 weeks. We showed that co-administration of cinnamaldehyde (10, 20, or 40 mg·kg-1·d-1, ig) for 6 weeks in F0 males prevented the depressive-like phenotypes of F1 male offspring. In addition, co-administration of cinnamaldehyde (20 mg·kg-1·d-1, ig) for 4 weeks significantly ameliorated depressive-like behaviors of chronic variable stress (CVS)-stimulated F1 offspring born to CMS mice. Notably, cinnamaldehyde had no reproductive toxicity, while positive drug fluoxetine showed remarkable reproductive toxicity. We revealed that CMS and CORT significantly reduced testis glucocorticoid receptor (GR) expression, and increased testis and sperm miR-190b expression in F0 depressive-like models. Moreover, pre-miR-190b expression was upregulated in testis of F0 males. The amount of GR on miR-190b promoter regions was decreased in testis of CORT-stimulated F0 males. Cinnamaldehyde administration reversed CORT-induced GR reduction in testis, miR-190b upregulation in testis and sperm, pre-miR-190b upregulation in testis, and the amount of GR on miR-190b promoter regions of F0 males. In miR-190b-transfected Neuro 2a (N2a) cells, we demonstrated that miR-190b might directly bind to the 3'-UTR of brain-derived neurotrophic factor (BDNF). In the hippocampus of F1 males of CORT- or CMS-induced depressive-like models, increased miR-190b expression was accompanied by reduced BDNF and GR, which were ameliorated by cinnamaldehyde. In conclusion, cinnamaldehyde is a potential intervening agent for intergenerational inheritance of depression, probably by regulating GR/miR-190b/BDNF pathway.

Effect of different phosphate binders on fibroblast growth factor 23 levels in patients with chronic kidney disease: a systematic review and meta-analysis of randomized controlled trials.

BACKGROUND: Serum intact fibroblast growth factor 23 (FGF23) levels are progressively increased in relation to the severity of kidney dysfunction. High serum intact FGF23 concentration is associated with the increased cardiovascular morbidity and mortality in patients with chronic kidney disease (CKD). Clinically, phosphate binders are commonly used to reduce serum intact FGF23 levels in CKD patients by lowering serum phosphate levels. It is not clear whether all kinds of phosphate binders can reduce serum intact FGF23 levels, or which kind of phosphate binders is more effective in reducing serum intact FGF23 levels in patients with CKD. The aim of this systematic review and meta-analysis was to compare the efficiency of different kinds of commonly used phosphate binders on serum intact FGF23 levels in patients with CKD. METHODS: Systematic searches were performed through PubMed, Cochrane Central Register of Controlled Trials and Embase from 1999 to 2020. We included the studies performed only in human subjects. All randomized clinical trials (RCTs) were included. Reviews, case reports, letters, commentaries, abstracts and unpublished articles were excluded. Risk of bias was assessed by the Cochrane Collaboration's tool. Random effect was performed in meta-analysis. Meta-regression was used to investigate heterogeneity. RESULTS: Of 1,895 articles, 15 RCTs (comprising 1,098 participants) were included. Common sources of bias were selection bias. Phosphate binders could reduce serum intact FGF23 levels in patients with CKD [standard mean difference (SMD) of total change in serum intact FGF23 levels was 0.91 PG/mL (95% confidence interval: 0.38 to 1.44 PG/mL]. Meta-regression explained 89.02% of heterogeneous sources, indicating that dietary phosphate intake could weaken the effect of phosphate binders on reducing serum intact FGF23 levels, and the effect of phosphate binders on reducing serum intact FGF23 levels in dialysis patients was better than that in early-to-middle CKD patients. DISCUSSION: Phosphate binders can effectively reduce serum intact FGF23 levels in CKD patients, and iron-based phosphate binders have better effect on reducing serum intact FGF23 levels than other phosphate binders.

Atractylodis rhizoma water extract attenuates fructose-induced glomerular injury in rats through anti-oxidation to inhibit TRPC6/p-CaMK4 signaling.

BACKGROUND: Atractylodis rhizoma, an aromatic herb for resolving dampness, is used to treat Kidney-related edema in traditional Chinese medicine for thousands years. This herb possesses antioxidant effect. However, it is not yet clear how Atractylodis rhizoma prevents glomerular injury through its anti-oxidation. PURPOSE: Based the analysis of Atractylodis rhizoma water extract (ARE) components and network pharmacology, this study was to explore whether ARE prevented glomerular injury via its anti-oxidation to inhibit oxidative stress-driven transient receptor potential channel 6 (TRPC6) and its downstream molecule calcium/calmodulin-dependent protein kinase IV (CaMK4) signaling. METHODS: Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used to analyze ARE components. Network pharmacology analysis was preliminarily performed. Male Sprague-Dawley rats were given 10% fructose drinking water (100 mL/d) for 16 weeks. ARE at 720 and 1090 mg/kg was orally administered to rats for the last 8 weeks. Hydrogen peroxide (H2O2) and malondialdehyde (MDA) level, and superoxide dismutase (SOD) activity in rat kidney cortex were detected, respectively. In rat glomeruli, redox-related factors forkhead box O3 (FoxO3), SOD2 and catalase (CAT), podocyte slit diaphragm proteins podocin and nephrin, cytoskeleton proteins CD2-associated protein (CD2AP) and α-Actinin-4, as well as TRPC6, p-CaMK4 and synaptopodin protein levels were analyzed by Western Blotting. SOD2 and CAT mRNA levels were detected by qRT-PCR. RESULTS: 36 components were identified in ARE. Among them, network pharmacology analysis indicated that ARE might inhibit kidney oxidative stress. Accordingly, ARE up-regulated nuclear FoxO3 expression, and then increased SOD2 and CAT at mRNA and protein levels in glomeruli of fructose-fed rats. It reduced H2O2 and MDA levels, and increased SOD activity in renal cortex of fructose-fed rats. Subsequently, ARE down-regulated TRPC6 and p-CaMK4, and up-regulated synaptopodin in glomeruli of fructose-fed rats. Furthermore, ARE increased podocin and nephrin, as well as CD2AP and α-Actinin-4, being consistent with its reduction of urine albumin-to-creatinine ratio and improvement of glomerular structure injury in this animal model. CONCLUSIONS: These results suggest that ARE may prevent glomerular injury in fructose-fed rats possibly by reducing oxidative stress to inhibit TRPC6/p-CaMK4 signaling and up-regulate synaptopodin expression. Therefore, ARE may be a promising drug for treating high fructose-induced glomerular injury in clinic.

Fabrication of supramolecular star-shaped amphiphilic copolymers for ROS-triggered drug release.

Star-shaped copolymers with branched structures can form unimolecular micelles with better stability than the micelles self-assembled from conventional linear copolymers. However, the synthesis of star-shaped copolymers with precisely controlled degree of branching (DB) suffers from complicated sequential polymerizations and multi-step purification procedures, as well as repeated optimizations of polymer compositions. The use of a supramolecular host-guest pair as the block junction would significantly simplify the preparation. Moreover, the star-shaped copolymer-based unimolecular micelle provides an elegant solution to the tradeoff between extracellular stability and intracellular high therapeutic efficacy if the association/dissociation of the supramolecular host-guest joint can be triggered by the biologically relevant stimuli. For this purpose, in this study, a panel of supramolecular star-shaped amphiphilic block copolymers with 9, 12, and 18 arms were designed and fabricated by host-guest complexations between the ring-opening polymerization (ROP)-synthesized star-shaped poly(ε-caprolactone) (PCL) with 3, 4, and 6 arms end-capped with ferrocene (Fc) (PCL-Fc) and the atom transfer radical polymerization (ATRP)-produced 3-arm poly(oligo ethylene glycol) methacrylates (POEGMA) with different degrees of polymerization (DPs) of 24, 30, 47 initiated by ß-cyclodextrin (ß-CD) (3Br-ß-CD-POEGMA). The effect of DB and polymer composition on the self-assembled properties of the five star-shaped copolymers was investigated by dynamic light scattering (DLS), transmission electron microscopy (TEM), and fluorescence spectrometery. Interestingly, the micelles self-assembled from 12-arm star-shaped copolymers exhibited greater stability than the 9- and 18-arm formulations. The potential of the resulting supramolecular star-shaped amphiphilic copolymers as drug carriers was evaluated by an in vitro drug release study, which confirmed the ROS-triggered accelerated drug release from the doxorubicin (DOX)-loaded supramolecular star-shaped micelles due to the oxidation-induced dissociation of ß-CD/Fc pair and the consequent loss of the colloidal stability of the star-shaped micelles. Studies of the delivery efficacy by an in vitro cytotoxicity study further indicated that higher DBs and longer hydrophilic arm compromised the therapeutic efficacy of the DOX-loaded supramolecular star-shaped micelles, resulting in significantly reduced cytotoxicity, as measured by increased IC50 value. Overall, our results revealed that the screening of hydrophilic block by DB and MW for an optimized star-shaped copolymer should balance the stability versus therapeutic efficacy tradeoff for a comprehensive consideration. Therefore, the 12-arm star-shaped copolymer with POEGMA30 is the best formulation tested.

A comparison study to investigate the effect of the drug-loading site on its delivery efficacy using double hydrophilic block copolymer-based prodrugs.

Polymeric delivery vehicles can improve the safety and efficacy of chemotherapy drugs by facilitating preferential tumor delivery. Double hydrophilic block copolymer (DHBC)-based prodrugs are considered as ideal candidates for drug delivery due to the elegant integration of benefits from both structures including polymeric prodrugs' superior protection and minimal premature drug release using covalent links and a DHBC-based "green" self-assembly strategy by a simple stimulus in a pure aqueous phase without the use of any organic solvent. Clearly, the location of drug molecules in the polymeric prodrugs has exerted a significant effect on their therapeutic efficiency. However, there has been no published data so far, to our knowledge, reporting the effect of drug-conjugated sites on its therapeutic efficacy, as well as some basic guidelines that can be followed to direct the future design of polymeric prodrugs. To this end, herein a thermo-sensitive DHBC, poly(N-(2-hydroxypropyl) methacrylamide)-b-poly(N-isopropyl acrylamide) (P(HPMA)-b-P(NIPAAm)), was designed and synthesized by successive reversible addition and fragmentation chain transfer (RAFT) polymerizations, and was chosen as a platform to clarify this issue. An anti-cancer drug, doxorubicin (DOX) was conjugated to the hydrophilic PHPMA block and the temperature-responsive P(NIPAAm) block, respectively, through a pH-liable hydrazone bond to fabricate two different types of polymeric prodrugs with the drug tethered to the micellar hydrophilic PHPMA shell or encapsulated within the hydrophobic P(NIPAAm) core upon temperature elevation above its lower critical solution temperature (LCST). A detailed comparison study was carried out to investigate which structure exhibits better properties and higher therapeutic efficacy in terms of micellar size, stability, cellular uptake, drug loading capacity, drug release behaviors and cell viability. The results showed the self-assembly of both DHBC-based prodrugs into well-dispersed spherical micelles with similar average hydrodynamic diameters (Dh) around 150 nm in phosphate buffer (PBS, pH 7.4) at 37 °C, but a higher drug loading content (DLC), and enhanced pH-mediated drug release, i.e., much accelerated drug release at pH 5.0, while slower at pH 7.4, as well as enhanced cytotoxicity when the drug was conjugated to the hydrophilic shell of the micelles. The guidelines obtained in this study are thus believed to direct the future design and development of polymeric prodrugs for efficient cancer therapy.

Fabrication of Dual-Redox Responsive Supramolecular Copolymers Using a Reducible ß-Cyclodextran-Ferrocene Double-Head Unit.

Self-assembly of amphiphilic block copolymers into well-defined nanostructures as drug delivery systems for the treatment of cancer has been a hot subject of research. However, sequential polymerizations synthesized amphiphilic block copolymers with covalent links suffered mainly from multistep synthesis and purification procedures as well as repeated optimization of polymer composition to form aggregates with well-defined structures. To overcome these drawbacks, supramolecular amphiphilic block copolymers with noncovalent links were developed to provide simplicity as required. Herein, we designed and prepared a reducible ß-cyclodextran (ß-CD)-ferrocene (Fc) double-head unit from which a dual-redox responsive supramolecular amphiphilic copolymer was fabricated together with a traditional polymer block through supramolecular induced polymerization. Typically, well-defined supramolecular micelles and vesicles were fabricated, respectively. Due to the integration of oxidation-sensitive noncovalent ß-CD/Fc connections and reduction-sensitive covalent disulfide bridges in the polymer backbone, the resulting supramolecular micelles and vesicles showed structural deformation and accelerated drug release in response to both intracellular reducing and oxidizing environments, thus, presenting a new platform for both reactive oxygen species (ROS) and glutathione (GSH)-triggered anticancer drug delivery.

Low temperature induces cryoinjury in mouse corneal endothelial cells by stimulating the Stk11-p53 signal pathway.

Cryoinjury, or injury caused by extremely low temperatures, can occur in corneal endothelial cells (CECs) and lead to visual impairment. However, the mechanism of cryoinjury in CECs is not clear. The Stk11­p53 signaling pathway regulates the proliferation and division of cells. Activity of the Stk11­p53 signaling pathway arrests the cell cycle at the G0/G1 phase and induces apoptosis. In this study, a mouse model of cryoinjury in CECs was used. Following injury, significant mouse CEC death and shedding were observed. In addition, the mRNA and protein levels of core factors from the Stk11­p53 signaling pathway (Stk11, p21 and p53) were elevated and Caspase­3 was activated following cryoinjury. In addition, chromatin immunoprecipitation revealed that Stk11 catalyzed p53 serine 15 phosphorylation, and the Stk11­p53 complex bound to the p21 promoter and stimulated gene transcription. Thus, the results of the present study suggest that cryoinjury leads to the damage and apoptosis of mouse CECs by activation of the Stk11­p53 signaling pathway, phosphorylation of p53 serine 15 and p21 gene transcription.

Ring reconstruction on a trichalcogenasumanene buckybowl: a facile approach to donor-acceptor-type [5-6-7] fused planar polyheterocycles.

The transformation of trichalcogenasumanene buckybowls into donor-acceptor-type [5-6-7] fused polyheterocycles is disclosed. The strategy involves a highly efficient ring-opening of the flanking benzene upon oxidation at room temperature, and facile ring closure by functional-group transformation. Crystallographic studies indicate that the resulting [5-6-7] fused polyheterocycles possess a planar conformation owing to the release of ring strain by expansion of one of the six-membered flanking rings to the seven-membered one. Additionally, the [5-6-7] fused polyheterocycles bear electron-withdrawing groups, which reduce the HOMO-LUMO energy gap, and display broad absorption bands extending to λ=590 nm. Consequently, these compounds show strong red emission with fluorescence quantum yields of up to 38 %.

Osteoblast inhibition by chemokine cytokine ligand3 in myeloma-induced bone disease.

BACKGROUND: Multiple myeloma is a hematologic malignancy characterized by the accumulation of monoclonal plasma cells in the bone marrow. A common manifestation of the disease is myeloma bone disease (MBD), which is caused by increased osteoclastic bone resorption and decreased bone formation. The chemokine cytokine ligand 3 (CCL3) is a pro-inflammatory protein and chemokine that stimulates osteoclasts in MBD. However, little is known about the effect of CCL3 on osteoblasts (OB). METHODS: The OBs are induced from patients with MBD and healthy donors, cultured in vitro, and identified by histochemistry. The effects of CCL3 and CCL3 antibody on the OBs in vitro are observed. The CCL3 receptor (CCR1), osteocalcin (OCN), runt-related transcription factor 2 (Runx2), and osterix (Osx) are detected using flow cytometry, enzyme-linked immunosorbent assay, and real-time PCR. RESULTS: Proliferation and osteogenic potential of the OB in patients with MBD are suppressed. Moreover, the CCR1 expression is significantly higher in patients with MBD than in normal controls. The OCN level, quantity of calcium nodules, and Runx2 and Osx levels decrease after CCL3 stimulation, which indicates that CCL3 inhibits OB function. Furthermore, CCL3 antibody partially restores OB activity through the upregulation of the OCN, Runx2, and Osx. CONCLUSIONS: CCL3 contributes to the OB/OC imbalance by inhibiting OB differentiation and function in MBD.