miRNA microarray profiling in patients with androgenic alopecia and the effects of miR-133b on hair growth.

OBJECTIVE: Androgenetic alopecia (AGA), a common alopecia, is often accompanied by abnormal expression of multiple miRNAs. This study aims to investigate abnormally expressed miRNAs in patients with AGA and their specific molecular mechanism. METHODS: miRNA microarray profiling and qRT-PCR validation were used to screen and verify abnormally expressed miRNAs in patients with AGA. Human hair follicles (HFs) were treated with different concentrations of dihydrotestosterone (DHT, 10-5, 10-6, 10-7 and 10-8 mol/L) for 10 days. The effects of DHT on HF growth, proliferation, and miRNA expression in cultured HFs were investigated using immunofluorescence staining and qRT-PCR. Moreover, human dermal papilla cells (HDPCs) were treated/transfected with a Wnt/ß-catenin pathway activator and/or miR-133b mimic, and then the CCK-8 assay was used to evaluate HDPC proliferation. qRT-PCR and Western blotting were used to measure the expression of Versican, ALP and ß-catenin RESULTS: miRNA microarray profiling identified 43 miRNAs that were significantly differentially expressed in AGA patients, and qRT-PCR verified that 8 miRNAs were significantly differentially expressed. The expression of miR-133b was abnormally high in AGA patients. DHT (10-5 mol/L) inhibited human HF growth and upregulated miR-133b expression, and DHT (10-7 mol/L) induced human HF growth and downregulated miR-133b expression. HDPC proliferation was inhibited, and the expression of ß-catenin was downregulated in the miR-133b mimic-transfected group compared with the control group (P < 0.05). Wnt/ß-catenin pathway activator treatment significantly promoted HDPC proliferation and upregulated the expression of ß-catenin (P < 0.05). In addition, the proliferation of HDPCs was not significantly different between the group cotreated with a Wnt/ß-catenin pathway activator and miR-133b mimic, and the control group (P > 0.05), but the expression of Versican and ALP was suppressed in the cotreatment group (P < 0.05) CONCLUSION: Our data indicated that patients with androgenic alopecia have specific miRNA expression profiles and that the abnormal expression of miR-133b may inactivate the Wnt/ß-catenin pathway and ultimately regulate hair growth.

Effects of chironomid larvae and Limnodrilus hoffmeisteri bioturbation on the distribution and flux of chromium at the sediment-water interface.

The impacts of chironomid larvae and the tubificid worm Limnodrilus hoffmeisteri on the distribution and flux of the heavy metal chromium (Cr) across the sediment-water interface were investigated with a 21-day laboratory microcosm experiment. The two studied species feature different bioturbation modes involving bioirrigation and upward bioconveyance. The Cr concentrations in the overlying water and pore water were measured and compared using treatments with bioturbation by a single species and by combinations of both species and a treatment with no organisms. The results indicated that both bioturbation modes significantly increased the Cr concentrations in the overlying water and pore water. The overlying water had lower Cr concentrations than the pore water. Little variation in the Cr concentrations was observed in the treatment without organisms. Both species enhanced the Cr flux from the pore water to the overlying water. The worm treatments had a great impact on the Cr concentration in the overlying water through intensive upward conveyance activity, while the chironomid larvae treatments exerted significant effects on the Cr variation in the pore water and Cr flux across the interface via bioirrigation activity. These findings reveal the importance of bioturbation in biogeochemical processes in freshwater ecosystems.

Activation of Wnt/ß-catenin signaling is involved in hair growth-promoting effect of 655-nm red light and LED in in vitro culture model.

Activation of the Wnt/ß-catenin signaling pathway plays an important role in hair follicle morphogenesis and hair growth. Recently, low-level laser therapy (LLLT) was evaluated for stimulating hair growth in numerous clinical studies, in which 655-nm red light was found to be most effective and practical for stimulating hair growth. We evaluated whether 655-nm red light + light-emitting diode (LED) could promote human hair growth by activating Wnt/ß-catenin signaling. An in vitro culture of human hair follicles (HFs) was irradiated with different intensities of 655-nm red light + LED, 21 h7 (an inhibitor of ß-catenin), or both. Immunofluorescence staining was performed to assess the expression of ß-catenin, GSK3ß, p-GSK3ß, and Lef1 in the Wnt/ß-catenin signaling. The 655-nm red light + LED not only enhanced hair shaft elongation, but also reduced catagen transition in human hair follicle organ culture, with the greatest effectiveness observed at 5 min (0.839 J/cm2). Additionally, 655-nm red light + LED enhanced the expression of ß-catenin, p-GSK3ß, and Lef1, signaling molecules of the Wnt/ß-catenin pathway, in the hair matrix. Activation of Wnt/ß-catenin signaling is involved in hair growth-promoting effect of 655-nm red light and LED in vitro and therefore may serve as an alternative therapeutic option for alopecia.

Molecular analysis of dominant species in Listeria monocytogenes-positive biofilms in the drains of food processing facilities.

Listeria monocytogenes exhibits symbiotic codependence with the dominant commensal bacteria, which may help it avoid being removed or inactivated by disinfectants in local environments. In this study, we investigated L. monocytogenes-positive biofilms at food production facilities, and the dominant bacterial species of the biofilms were identified to determine the properties of the microbiological background. For this purpose, the ISO 11290 method was used for the detection and isolation of L. monocytogenes, and the species were further identified based on 16S rRNA and hly genes. 16S rRNA gene-based cloning, terminal restriction fragment length polymorphism, and denaturing gradient gel electrophoresis were combined to evaluate the dominant bacteria of the drain biofilms. Out of 100 drain samples, 8 were naturally contaminated with L. monocytogenes. Three molecular methods consistently showed that Pseudomonas psychrophila, Pseudomonas sp., and Klebsiella oxytoca were dominant species in 3Q, 5Q, and 6Q samples; Aeromonas hydrophila and Klebsiella sp. were significantly dominant in 1-2, 1-3, and 3-2 samples; A. hydrophila and K. oxytoca were dominant in the 2-3 sample; and A. hydrophila and Pseudomonas sp. were prominent in the 3-3 sample. Different biofilms from the same plant shared common bands, suggesting that similar bacteria can be found and can be dominant in different biofilms. This study provides a better understanding of the dominant compositions in these bacterial communities. Further studies to determine the mechanism of co-culture with L. monocytogenes will be of critical importance in predicting effective disinfection strategies.

Laser-Activatable In Situ Vaccine Enhances Cancer-Immunity Cycle.

The immune response in cancer reflects a series of carefully regulated events; however, current tumor immunotherapies typically address a single key aspect to enhance anti-tumor immunity. In the present study, a nanoplatform (Fe3 O4 @IR820@CpG)-based immunotherapy strategy that targets the multiple key steps in cancer-immunity cycle is developed: 1) promotes the release of tumor-derived proteins (TDPs), including tumor-associated antigens and pro-immunostimulatory factors), in addition to the direct killing effect, by photothermal (PTT) and photodynamic therapy (PDT); 2) captures the released TDPs and delivers them, together with CpG (a Toll-like receptor 9 agonist) to antigen-presenting cells (APCs) to promote antigen presentation and T cell activation; 3) enhances the tumor-killing ability of T cells by combining with anti-programmed death ligand 1 antibody (α-PD-L1), which collectively advances the outstanding of the anti-tumor effects on colorectal, liver and breast cancers. The broad-spectrum anti-tumor activity of Fe3 O4 @IR820@CpG with α-PD-L1 demonstrates that optimally manipulating anti-cancer immunity not singly but as a group provides promising clinical strategies.

A RIPK3-independent role of MLKL in suppressing parthanatos promotes immune evasion in hepatocellular carcinoma.

Mixed lineage kinase domain-like (MLKL) is widely accepted as an executioner of necroptosis, in which MLKL mediates necroptotic signaling and triggers cell death in a receptor-interacting protein kinase 3 (RIPK3)-dependent manner. Recently, it is increasingly noted that RIPK3 is intrinsically silenced in hepatocytes, raising a question about the role of MLKL in hepatocellular carcinoma (HCC). This study reports a previously unrecognized role of MLKL in regulating parthanatos, a programmed cell death distinct from necroptosis. In HCC cells with intrinsic RIPK3 deficiency, knockout of MLKL impedes the orthotopic tumor growth, activates the anti-tumor immune response and enhances the therapeutic effect of immune checkpoint blockade in syngeneic HCC tumor models. Mechanistically, MLKL is required for maintaining the endoplasmic reticulum (ER)-mitochondrial Mg2+ dynamics in HCC cells. MLKL deficiency restricts ER Mg2+ release and mitochondrial Mg2+ uptake, leading to ER dysfunction and mitochondrial oxidative stress, which together confer increased susceptibility to metabolic stress-induced parthanatos. Importantly, pharmacological inhibition of poly(ADP-ribose) polymerase to block parthanatos restores the tumor growth and immune evasion in MLKL-knockout HCC tumors. Together, our data demonstrate a new RIPK3-independent role of MLKL in regulating parthanatos and highlight the role of MLKL in facilitating immune evasion in HCC.

The N-Terminal α-Helix of Potato Virus X-Encoded RNA-Dependent RNA Polymerase Is Required for Membrane Association and Multimerization.

Positive-sense single-stranded RNA viruses replicate in virus-induced membranous organelles for maximum efficiency and immune escaping. The replication of potato virus X (PVX) takes place on the endoplasmic reticulum (ER); however, how PVX-encoded RNA-dependent RNA polymerase (RdRp) is associated with the ER is still unknown. A proline-kinked amphipathic α-helix was recently found in the MET domain of RdRp. In this study, we further illustrate that the first α-helix of the MET domain is also required for ER association. Moreover, we found that the MET domain forms multimers on ER and the first α-helix is essential for multimerization. These results suggest that the RdRp of PVX adopts more than one hydrophobic motif for membrane association and for multimerization.

Fine Mapping the Soybean Mosaic Virus Resistance Gene in Soybean Cultivar Heinong 84 and Development of CAPS Markers for Rapid Identification.

Heinong 84 is one of the major soybean varieties growing in Northeast China, and is resistant to the infection of all strains of soybean mosaic virus (SMV) in the region including the most prevalent strain, N3. However, the resistance gene(s) in Heinong 84 and the resistant mechanism are still elusive. In this study, genetic and next-generation sequencing (NGS)-based bulk segregation analysis (BSA) were performed to map the resistance gene using a segregation population from the cross of Heinong 84 and a susceptible cultivar to strain N3, Zhonghuang 13. Results show that the resistance of Heinong 84 is controlled by a dominant gene on chromosome 13. Further analyses suggest that the resistance gene in Heinong 84 is probably an allele of Rsv1. Finally, two pairs of single-nucleotide-polymorphism (SNP)-based primers that are tightly cosegregated with the resistance gene were designed for rapidly identifying resistant progenies in breeding via the cleaved amplified polymorphic sequence (CAPS) assay.

Effects of land use on slope runoff and soil loss in the Loess Plateau of China: A meta-analysis.

In the Loess Plateau, due to the inappropriate vegetation restoration mode, large areas of artificially restored vegetation began to degrade, thus the optimization of vegetation allocation has become an urgent necessity. The main purpose of this study was to identify and evaluate slope runoff and soil loss rates, and to review all of the plot-scale studies in the Chinese Loess Plateau, by meta-analysis. Based on data collected from the runoff plot, the effect of land use on annual runoff and annual soil loss under natural rainfall conditions was analyzed. The optimization of land use in different climatic regions of the Loess Plateau was evaluated. The plot database contained 55 plot measuring sites in the Loess Plateau, which included 461 runoff plots and 535 soil loss plots. Bare soil was found to have the highest average annual runoff (58.57 mm·yr-1) and annual soil loss (122.06 t·ha-1·yr-1). Natural grassland and mixed forest had the lowest annual runoff (<15 mm·yr-1) and annual soil loss (<20 t·ha-1·yr-1), exhibiting a better effect of soil and water conservation when the precipitation was <200 mm and >600 mm, respectively. When the precipitation was 400-600 mm, shrubland showed the lowest mean annual runoff (21.36 mm·yr-1) and annual soil loss (13.36 t·ha-1·yr-1), which conducive to reducing water and sediment. Therefore, shrubland could be selected as the recovery vegetation type in the semi-humid climatic region. Land-use types determined the relationship between annual soil loss and annual runoff with plot length and slope gradient. These results enabled the assessment of the impact of land-use change on water erosion, providing a basis for formulating soil and water conservation management programs.

Dihydrotestosterone Regulates Hair Growth Through the Wnt/ß-Catenin Pathway in C57BL/6 Mice and In Vitro Organ Culture.

Dihydrotestosterone (DHT) is the most potent androgen that regulates hair cycling. Hair cycling involves cross-talk between the androgen and Wnt/ß-catenin pathways. However, how DHT regulates hair follicle (HF) growth through the Wnt/ß-catenin pathway has not been well investigated. This study aimed to investigate the roles of DHT in hair growth in vivo and in vitro. Human scalp HFs were treated with different concentrations of DHT (10-5, 10-6, 10-7, 10-8, and 10-9 mol/L) for 10 days. The effects of DHT on hair shaft elongation, the proliferation of hair matrix cells, and the levels of ß-catenin, GSK-3ß, and phosphorylated GSK-3ß (ser9) were evaluated in the cultured HFs. The effects of DHT were further investigated in C57BL/6 mice. Moreover, the growth of cultured human HFs was observed after interfering with the ß-catenin pathway through inhibitors or activators in the presence or absence of DHT. We found that different concentrations of DHT had different effects on human HFs in vitro and C57BL/6 mice. At 10-6 mol/L, DHT inhibited HF growth and ß-catenin/p-GSK-3ß expression, whereas 10-7 mol/L DHT induced HF growth and ß-catenin/p-GSK-3ß expression. In addition, a ß-catenin inhibitor (21H7) inhibited HF growth in vitro, while a ß-catenin activator (IM12) promoted HF growth in vitro and antagonized the inhibition of HFs by high levels of DHT. These results suggest that DHT plays a pivotal role in region-specific hair growth, which may be related to the Wnt/ß-catenin pathway.

Design, synthesis and biological evaluation of tetrahydronaphthyridine derivatives as bioavailable CDK4/6 inhibitors for cancer therapy.

CDK4/6 pathway is an attractive chemotherapeutic target for antitumor drug discovery and development. Herein, we reported the structure-based design and synthesis of a series of novel tetrahydronaphthyridine analogues as selective CDK4/6 inhibitors. Compound 5 was identified as a hit and then systematically structure optimization study was conducted. These efforts led to compound 28, which exhibited excellent in vitro potencies against CDK4/6 enzymatic activity with high selectivity over CDK1, and against Colo-205 cell growth. The compound demonstrated favorable in vitro metabolic and robust mice pharmacokinetic properties. In Colo-205 xenograft models, compound 28 showed potent tumor growth inhibition with acceptable toxic effects, which could serve as a novel anticancer agent for further preclinical study.

ß-Catenin is involved in oleanolic acid-dependent promotion of proliferation in human hair matrix cells in an in vitro organ culture model.

Oleanolic acid (OA), a pentacyclic triterpenoid compound which can be found in >1600 plants, has been shown to promote hair growth. To study the mechanisms of OA on hair growth, we investigated hair follicle (HF) growth on four different concentration OA using human hair follicle organ culture model. We found that HFs treated with 1 or 10µg/mL OA showed statistically enhanced elongation of the hair shaft and anagen-like stage. Moreover, higher positive rate of Ki-67, a matrix cellular marker of proliferation, was detected in the same groups treated with 1 or 10µg/mL than those treated with vehicle. We further demonstrated that ß-catenin, a key Wnt signaling transducer, was highly expressed in the OA treated groups using immunofluorescence stain assay. These results suggest that OA may promote human hair growth by stimulating hair matrix cell proliferation through the Wnt/ß-catenin pathway.