A highly sensitive Lock-Cas12a biosensor for detection and imaging of miRNA-21 in breast cancer cells.

The expression levels of microRNA (miRNA) vary significantly in correlation with the occurrence and progression of cancer, making them valuable biomarkers for cancer diagnosis. However, their quantitative detection faces challenges due to the high sequence homology, low abundance and small size. In this work, we established a strand displacement amplification (SDA) approach based on miRNA-triggered structural "Lock" nucleic acid ("Lock" DNA), coupled with the CRISPR/Cas12a system, for detecting miRNA-21 in breast cancer cells. The "Lock" DNA freed the CRISPR-derived RNA (crRNA) from the dependence on the target sequence and greatly facilitated the extended detection of different miRNAs. Moreover, the CRISPR/Cas12a system provided excellent amplification ability and specificity. The designed biosensor achieved high sensitivity detection of miRNA-21 with a limit of detection (LOD) of 28.8 aM. In particular, the biosensor could distinguish breast cancer cells from other cancer cells through intracellular imaging. With its straightforward sequence design and ease of use, the Lock-Cas12a biosensor offers significant advantages for cell imaging and early clinical diagnosis.

Integrative transcriptome and metabolome analysis reveals the discrepancy in the accumulation of active ingredients between Lycium barbarum cultivars.

MAIN CONCLUSION: The combined analysis of transcriptome and metabolome provided molecular insight into the dynamics of multiple active ingredients biosynthesis and accumulation across different cultivars of Lycium barbarum. Lycium barbarum L. has a high concentration of active ingredients and is well known in traditional Chinese herbal medicine for its therapeutic properties. However, there are many Lycium barbarum cultivars, and the content of active components varies, resulting in inconsistent quality between Lycium barbarum cultivars. At present, few research has been conducted to reveal the difference in active ingredient content among different cultivars of Lycium barbarum at the molecular level. Therefore, the transcriptome of 'Ningqi No.1' and 'Qixin No.1' during the three development stages (G, T, and M) was constructed in this study. A total of 797,570,278 clean reads were obtained. Between the two types of wolfberries, a total of 469, 2394, and 1531 differentially expressed genes (DEGs) were obtained in the 'G1 vs. G10,' 'T1 vs. T10,' and 'M1 vs. M10,' respectively, and were annotated with Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) orthology identifiers. Using these transcriptome data, most DEGs related to the metabolism of the active ingredients in 'Ningqi No.1' and 'Qixin No.1' were identified. Moreover, a widely targeted metabolome analysis of the metabolites of 'Ningqi 1' and 'Qixin 1' fruits at the maturity stage revealed 1,135 differentially expressed metabolites (DEMs) in 'M1 vs. M10,' and many DEMs were associated with active ingredients such as flavonoids, alkaloids, terpenoids, and so on. We further quantified the flavonoid, lignin, and carotenoid contents of the two Lycium barbarum cultivars during the three developmental stages. The present outcome provided molecular insight into the dynamics of multiple active ingredients biosynthesis and accumulation across different cultivars of Lycium barbarum, which would provide the basic data for the formation of Lycium barbarum fruit quality and the breeding of outstanding strains.

Polyvinyl alcohol/carboxymethyl chitosan-based hydrogels loaded with taxifolin liposomes promote diabetic wound healing by inhibiting inflammation and regulating autophagy.

With the improvement of modern living standards, the challenge of diabetic wound healing has significantly impacted the public health system. In this study, our objective was to enhance the bioactivity of taxifolin (TAX) by encapsulating it in liposomes using a thin film dispersion method. Additionally, polyvinyl alcohol/carboxymethyl chitosan-based hydrogels were prepared through repeated freeze-thawing. In vitro and in vivo experiments were conducted to investigate the properties of the hydrogel and its effectiveness in promoting wound healing in diabetic mice. The results of the experiments revealed that the encapsulation efficiency of taxifolin liposomes (TL) was 89.80 ± 4.10 %, with a drug loading capacity of 17.58 ± 2.04 %. Scanning electron microscopy analysis demonstrated that the prepared hydrogels possessed a porous structure, facilitating gas exchange and the absorption of wound exudates. Furthermore, the wound repair experiments in diabetic mice showed that the TL-loaded hydrogels (TL-Gels) could expedite wound healing by suppressing the inflammatory response and promoting the expression of autophagy-related proteins. Overall, this study highlights that TL-Gels effectively reduce wound healing time by modulating the inflammatory response and autophagy-related protein expression, thus offering promising prospects for the treatment of hard-to-heal wounds induced by diabetes.

Superalkali-alkaline earthide ion pairs of δ+(AM-HMHC)-AM'δ- (AM = Li, Na and K; AM' = Be, Mg and Ca) possessing large NLO responses and excellent electronic stabilities and alkalide characteristics: a DFT study.

To identify superalkali-alkaline earthide ion pairs, it's theoretically shown that, as a novel class of excess electron superalkali compounds, both chair and boat forms of (AM-HMHC)-AM' (AM = Li, Na, and K; AM' = Be, Mg, and Ca; HMHC = 1,4,7,10,13,16-hexamethyl-1,4,7,10,13,16-hexaazacyclooctadecane) are good candidates. An attractive superalkali-alkaline earthide ion pair in δ+(AM-HMHC)-AM'δ- is firstly exhibited, which possesses alkaline-earthide characteristics and nonlinear optical response superior to similar M+(calix[4]pyrrole)M'- (M = Li, Na, and K; M' = Be, Mg, and Ca) with high stability. The electronic and vibrational second order hyperpolarizabilities and the frequency-dependent first hyperpolarizabilities of δ+(AM-HMHC)-AM'δ- are presented. For each pair of (AM-HMHC)-AM', the boat conformation is preferred to its chair one in the case of Hyper-Rayleigh scattering response (ßHRS). These alkaline earthides suggest prominently high ßHRS up to 2.59 × 104 a.u. (boat forms of δ+(Na-HMHC)-Caδ-). We expect that this work will inspire the preparation and characterization of these new alkaline earthides as high-performance NLO materials.

Transcriptional deciphering of the metabolic pathways associated with the bioactive ingredients of wolfberry species with different quality characteristics.

BACKGROUND: Wolfberry is rich in carotenoids, flavonoids, vitamins, alkaloids, betaines and other bioactive ingredients. For over 2,000 years, wolfberry has been used in China as a medicinal and edible plant resource. Nevertheless, the content of bioactive ingredients varies by cultivars, resulting in uneven quality across wolfberry cultivars and species. To date, research has revealed little about the underlying molecular mechanism of the metabolism of flavonoids, carotenoids, and other bioactive ingredients in wolfberry. RESULTS: In this context, the transcriptomes of the Lycium barbarum L. cultivar 'Ningqi No. 1' and Lycium chinense Miller were compared during the fruit maturity stage using the Illumina NovaSeq 6000 sequencing platform, and subsequently, the changes of the gene expression profiles in two types of wolfberries were analysed. In total, 256,228,924 clean reads were obtained, and 8817 differentially expressed genes (DEGs) were identified, then assembled by Basic Local Alignment Search Tool (BLAST) similarity searches and annotated using Gene Ontology (GO), Clusters of Orthologous Groups of proteins (KOG), and the Kyoto Encyclopedia of Genes and Genomes (KEGG). By combining these transcriptome data with data from the PubMed database, 36 DEGs related to the metabolism of bioactive ingredients and implicated in the metabolic pathway of carotenoids, flavonoids, terpenoids, alkaloids, vitamins, etc., were identified. In addition, among the 9 differentially expressed transcription factors, LbAPL, LbPHL11 and LbKAN4 have raised concerns. The protein physicochemical properties, structure prediction and phylogenetic analysis indicated that LbAPL and LbPHL11 may be good candidate genes involved in regulating the flavonoid metabolism pathway in wolfberry. CONCLUSIONS: This study provides preliminary evidence for the differences in bioactive ingredient content at the transcription level among different wolfberry species, as well as a research and theoretical basis for the screening, cloning and functional analysis of key genes involved in the metabolism of bioactive ingredients in wolfberry.

Enhanced hydrogen evolution reaction via regulating the adsorbability between 2D CoO nanosheets and CC substrate.

In recent years, bifunctional electrocatalysts, nanomaterials directly grown on the substrate for application towards the hydrogen evolution reaction (HER), have become of interest for sustainable and clean energy technologies. However, the influence of interfacial interactions between the electrode materials and substrate on device performance remains unclear and is rarely investigated. Herein, we report two-dimensional (2D) CoO nanosheets grown on carbon cloth (CC) (2D CoO/CC) to construct a hybrid electrocatalyst with a seamlessly conductive network. By a series of structure analyses, we recommend that the CoO nanosheets and CC are connected via adsorption. The 2D CoO/CC nanosheets show superior HER performance to the commercial Pt/C and CoO(aq.)/CC nanosheets, including onset potentials of 2 mV, low overpotential of 22 mV at 10 mA cm-2 and Tafel slope of 37 mV dec-1. The results of density functional theory (DFT) calculations reveal that the adsorbability plays an important role in determining the performance of the electrocatalysts for the HER. This work provides a new insight into the interfacial interactions between the electrode material and the substrate in electrochemical devices, and paves the way for the rational design and construction of high-performance electrochemical devices for practical energy applications.

Microenvironment-Responsive Metal-Phenolic Nanozyme Release Platform with Antibacterial, ROS Scavenging, and Osteogenesis for Periodontitis.

Periodontitis is a chronic inflammatory disease deriving from dental plaque, characterized by the excessive accumulation of reactive oxygen species (ROS), matrix metalloproteinase (MMP) and other substances, resulting in the destruction of periodontal tissues. At present, the main therapeutic modalities, such as local mechanical debridement and antibiotic delivery, are not only difficult to solve the intractable bacterial biofilm effectively but also tricky to ameliorate the excessive inflammatory response as well as regenerate the impaired periodontal tissues. Herein, we have proposed the TM/BHT/CuTA hydrogel system formed by the self-assembly of the copper-based nanozyme (copper tannic acid coordination nanosheets, CuTA NSs) and the triglycerol monostearate/2,6-di-tert-butyl-4-methylphenol (TM/BHT) hydrogel. The negatively charged TM/BHT/CuTA can retain at the inflammation sites with a positive charge through electrostatic adsorption and hydrolyze in response to the increasing MMP of periodontitis, realizing the on-demand release of the CuTA nanozyme. The released CuTA nanozyme has antibacterial and antiplaque properties. Meanwhile, as a metal-phenolic nanozyme, it can scavenge multiple ROS by simulating the cascade process of superoxide dismutase (SOD) and catalase (CAT). Further, the CuTA nanozyme can modulate the macrophage polarization from M1 phenotype to M2 phenotype through the Nrf2/NF-κB pathway, which reduces the pro-inflammatory cytokines, increases the anti-inflammatory cytokines, and promotes the expression of osteogenetic genes successively, thus relieving the inflammation and accelerating the tissue regeneration of periodontitis. Altogether, this multifunctional nanozyme on-demand release platform (TM/BHT/CuTA) provides a desirable strategy for the treatment of periodontitis.

Human umbilical cord mesenchymal stem cells (hUC-MSCs) alleviate paclitaxel-induced spermatogenesis defects and maintain male fertility.

Chemotherapeutic drugs can cause reproductive damage by affecting sperm quality and other aspects of male fertility. Stem cells are thought to alleviate the damage caused by chemotherapy drugs and to play roles in reproductive protection and treatment. This study aimed to explore the effects of human umbilical cord mesenchymal stem cells (hUC-MSCs) on alleviating paclitaxel (PTX)-induced spermatogenesis and male fertility defects. An in vivo PTX-induced mice model was constructed to evaluate the reproductive toxicity and protective roles of hUC-MSCs in male fertility improvement. A 14 day PTX treatment regimen significantly attenuated mice spermatogenesis and sperm quality, including affecting spermatogenesis, reducing sperm counts, and decreasing sperm motility. hUC-MSCs treatment could significantly improve sperm functional indicators. Mating experiments with normal female mice and examination of embryo development at 7.5 days post-coitum (dpc) showed that hUC-MSCs restored male mouse fertility that was reduced by PTX. In IVF experiments, PTX impaired sperm fertility and blastocyst development, but hUC-MSCs treatment rescued these indicators. hUC-MSCs' protective role was also displayed through the increased expression of the fertility-related proteins HSPA2 and HSPA4L in testes with decreased expression in the PTX-treated group. These changes might be related to the PTX-induced decreases in expression of the germ cell proliferation protein PCNA and the meiosis proteins SYCP3, MLH1, and STRA8, which were restored after hUC-MSCs treatment. In the PTX-treated group, the expression of testicular antioxidant proteins SIRT1, NRF2, CAT, SOD1, and PRDX6 was significantly decreased, but hUC-MSCs could maintain these expressions and reverse PTX-related increases in BAX/BCL2 ratios. hUC-MSCs may be a promising agent with antioxidant and anti-apoptosis characteristics that can maintain sperm quality following chemotherapy treatment.

[Comparative analysis of differentially expressed genes for biosynthesis of active ingredients in fruits of different cultivars of Lycium barbarum L. based on transcriptome sequencing].

To explore the differentially expressed genes (DEGs) related to biosynthesis of active ingredients in wolfberry fruits of different varieties of Lycium barbarum L. and reveal the molecular mechanism of the differences of active ingredients, we utilized Illumina NovaSeq 6000 high-throughput sequencing technology to conduct transcriptome sequencing on the fruits of 'Ningqi No.1' and 'Ningqi No.7' during the green fruit stage, color turning stage and maturity stage. Subsequently, we compared the profiles of related gene expression in the fruits of the two varieties at different development stages. The results showed that a total of 811 818 178 clean reads were obtained, resulting in 121.76 Gb of valid data. There were 2 827, 2 552 and 2 311 DEGs obtained during the green fruit stage, color turning stage and maturity stage of 'Ningqi No. 1' and 'Ningqi No. 7', respectively, among which 2 153, 2 050 and 1 825 genes were annotated in six databases, including gene ontology (GO), Kyoto encyclopedia of genes and genomes (KEGG) and clusters of orthologous groups of proteins (KOG). In GO database, 1 307, 865 and 624 DEGs of green fruit stage, color turning stage and maturity stage were found to be enriched in biological processes, cell components and molecular functions, respectively. In the KEGG database, the DEGs at three developmental stages were mainly concentrated in metabolic pathways, biosynthesis of secondary metabolites and plant-pathogen interaction. In KOG database, 1 775, 1 751 and 1 541 DEGs were annotated at three developmental stages, respectively. Searching the annotated genes against the PubMed database revealed 18, 26 and 24 DEGs related to the synthesis of active ingredients were mined at the green fruit stage, color turning stage and maturity stage, respectively. These genes are involved in carotenoid, flavonoid, terpenoid, alkaloid, vitamin metabolic pathways, etc. Seven DEGs were verified by RT-qPCR, which showed consistent results with transcriptome sequencing. This study provides preliminary evidences for the differences in the content of active ingredients in different Lycium barbarum L. varieties from the transcriptional level. These evidences may facilitate further exploring the key genes for active ingredients biosynthesis in Lycium barbarum L. and analyzing their expression regulation mechanism.

Development and characterization of a new potato starch/watermelon peel pectin composite film loaded with TiO2 nanoparticles and microencapsulated Lycium barbarum leaf flavonoids and its use in the Tan mutton packaging.

Reinforced edible film with active nanoparticles has been in increasing demand as a new technology to improve the quality and extend the shelf-life of muscle foods. The study aimed to fabricate and characterize a novel potato starch (Pst)/watermelon peel pectin (Wpp) composite film with the microencapsulated Lycium barbarum leaf flavonoids (MLF) and nano-TiO2 (Pst/Wpp/MLF/TiO2) and further apply the film in Tan mutton preservation. The moisture content, thickness and water vapor permeability (WVP) of the composite film were relatively increased with increasing the percentage of MLF, while nano-TiO2 had slight influence on the thickness, but leaded to a significantly decreased the moisture content and WVP. Also, the SEM images showed that the roughness and porosity were created on the film surface by adding MLF and nano-TiO2. FTIR revealed electrostatic and hydrogen bond interactions between the components in the film system. Meanwhile, MLF and nano-TiO2 effectively enhanced the mechanical strength, UV-barrier, controlled-release, thermal stability, antimicrobial and antioxidation properties of the Pst/Wpp film. Also, the composite film containing MLF and nano-TiO2 significantly inhibited the growth of microorganisms and chemical deterioration of mutton samples, which suggested that such film has potential as a prospective active packaging for preserving Tan mutton.

Melatonin improves the ability of spermatozoa to bind with oocytes in the mouse.

CONTEXT AND AIMS: Melatonin is a powerful antioxidant regulating various biological functions, including alleviating male reproductive damage under pathological conditions. Here, we aim to analyse the effect of melatonin on normal male reproduction in mice. METHODS: Male mice received an intraperitoneal injection of melatonin (10mg/kg body weight) for 35 consecutive days. The testis and epididymis morphology, and epididymal sperm parameters were examined. PCNA, HSPA2, SYCP3, ZO-1 and CYP11A1 expressions in epididymis or testis were detected by immunohistochemistry or Western blotting. Male fertility was determined by in vivo and in vitro fertilisation (IVF) experiments. The differentially expressed sperm proteins were identified by proteomics. KEY RESULTS: No visible structural changes and oxidative damage in the testis and epididymis, and no significant side effects on testis weight, testosterone levels, sperm motility, and sperm morphology were observed in the melatonin-treatment group compared with the control group. Spermatogenesis-related molecules of PCNA, SYCP3, ZO-1, and CYP11A1 showed no significant differences in melatonin-treated testis. However, PCNA and HSPA2 increased their expressions in the epididymal initial segments in the melatonin-treatment group. Normal sperm fertilisation, two-cell and blastocyst development were observed in the melatonin-treated group, but melatonin significantly enhanced the sperm binding ability characterised as more sperm binding to one oocyte (control 7.2±1.3 versus melatonin 11.8±1.5). Sperm proteomics demonstrated that melatonin treatment enhanced the biological process of cell adhesion in sperm. CONCLUSIONS AND IMPLICATIONS: This study suggests that melatonin can promote sperm maturation and sperm function, providing important information for further research on the physiological function and protective effect of melatonin in male reproduction.

In vivo study revealed pro-tumorigenic effect of CMTM3 in hepatocellular carcinoma involving the regulation of peroxisome proliferator-activated receptor gamma (PPARγ).

PURPOSE: To clarify the ambiguity of the function of CMTM3 in the development of hepatocellular carcinoma (HCC) and explore its molecular mechanism. METHODS: The Cmtm3-KO C57BL/6 mouse strain was established using CRISPR-Cas9. Acute liver damage and HCC models were induced by peritoneal injection of 100 or 25 mg/kg.BW N-Nitrosodiethylamine (DEN) to male mice. Liver function and histology were evaluated by blood serum levels of AST and ALT, and HE staining. Gene and protein expression in liver tissues was investigated by RNA-seq, RT-qPCR, Western blotting, immunohistochemistry, and immunofluorescence. Protein-protein interactions were studied by STRING and topological measures. The mRNA expression of CMTM3 and PPARs and patient survival were analyzed using the UALCAN database. RESULTS: Global knockout of Cmtm3 in KO mice was successfully confirmed. Cmtm3 knockout alleviated DEN-induced acute damage to liver histological integrity and liver function, reduced DNA damage and apoptosis, and also caused a significantly reduced number (WT: 8.7 ± 5.5 vs. KO: 2.7 ± 3.1, P = 0.0394) and total size of tumors (WT: 130.9 ± 181.8 mm2 vs. KO: 9.3 ± 11.5 mm2, P = 0.026) in the liver. Mechanistically, Cmtm3 knockout resulted in reduced expression and inactivation of Pparγ and its downstream lipid metabolism genes (e.g. Adipoq) upon DEN intoxication. CMTM3 and PPARγ were both overexpressed in HCC, and higher levels of both genes were associated with worse overall survival of HCC patients. CONCLUSION: This study clarified the pro-tumorigenesis role of CMTM3 in HCC in vivo, possibly through the upregulation of PPARγ and activation of the PPAR pathway.

Protective effect of melatonin on alleviating early oxidative stress induced by DOX in mice spermatogenesis and sperm quality maintaining.

Doxorubicin (DOX) is an effective chemotherapy drug, but its clinical use has adverse effects on male reproduction. However, there are few studies about the specific biological processes related to male reproduction or strategies for improving fertility protection. In this paper, we examined the effects of DOX on spermatogenesis and sperm function, and tested the possible protective role of melatonin (MLT) against DOX's reproductive toxicity. DOX-treated mice showed signs of significantly impaired spermatogenesis, including vacuolated epithelial cells, decreased testis weights, and lowered sperm counts and motility. DOX also reduced germ cell proliferation (PCNA) and meiosis-related proteins (SYCP3), but this effect could be partially improved with MLT administration. HSPA2 expression was maintained, which indicated that although MLT did not improve sperm motility, it did have a significant protective effect on elongated sperm. IVF results showed that MLT could partially promote two-cell and blastocyte development that was restricted by DOX. MLT reversed DOX-driven changes in the testes, including the antioxidant indices of SOD1, CAT and PRDX6, and the apoptotic indices of BAX and Caspase3. These results suggest that MLT effectively prevents DOX-induced early reproductive toxicity, and increase our understanding of the molecular mechanisms underlying DOX's effects on male reproduction and the protective mechanism of MLT.

An injectable and self-healing hydrogel with antibacterial and angiogenic properties for diabetic wound healing.

The treatment of diabetic wounds remains a global challenge. Compared with traditional wound dressings, there are higher requirements of antibacterial, anti-inflammatory and pro-angiogenic effects in diabetic wound dressings. Furthermore, it is desirable for dressings to self-adapt to wounds with different morphologies without extra processes and stably (suitable adhesive and self-healing abilities) provide a conducive environment for wound healing. Herein, we construct an injectable and self-healing hydrogel through the combination of chitosan (CS) and metal ions to efficiently improve infected and diabetic wound healing. Benefiting from the amino and hydroxy groups, the CS molecular chains are cross-linked with silver ions (Ag+) and copper ions (Cu2+) to promote the formation of the CS-Ag-Cu hydrogel, which releases Ag+ (an antibacterial agent) and Cu2+ (an angiogenic agent) over a prolonged period. Moreover, the hydrogel possesses appropriate adhesive ability, good water absorption ability, antibacterial capability and biocompatibility according to in vitro investigations. In vivo experimental results further prove that the CS-Ag-Cu hydrogel can dramatically accelerate tissue repair in a Staphylococcus aureus (S. aureus)-infected skin incision model in normal rats and diabetic wounds.

Can a molecular switch exist in both superalkali electride and superalkalide forms?

To combine both electride and alkalide characteristics in one molecular switch, it is shown herein that the phenalenyl radical and the M3 ring (M3-PHY, M = Li, Na, and K) stacked with parallel and vertical geometries are good candidates. The former geometry is the superalkali electride e-⋯M3+-PHY while the latter geometry is the superalkalide Mδ--M2(1-δ)+-PHY-. The superalkalide Mδ--M2(1-δ)+-PHY- may isomerize to the superalkali electride e-⋯M3+-PHY (M = Li, Na, and K) using suitable long-wavelength irradiation, while the latter may isomerize to the former with suitable short-wavelength irradiation. Also, applying suitable oriented external electric fields can drive the superalkalide Mδ-M2(1-δ)+-PHY- to change into the superalkali electride e-⋯M3+-PHY (M = Li, Na, and K). The differences in the static and dynamic first hyperpolarizability (ß0) values between them were also studied.

A graphene-Ag based near-infrared defined accurate anti-scarring strategy for ocular glaucoma surgery.

Excessive fibrosis is the major factor in the failure of glaucoma filtration surgery. So far, the dominant approach for inhibiting fibrosis is the use of an antimetabolite drug, but the complications it causes, such as filtering bleb leakage, bacterial endophthalmitis and ocular hypotony, are also inevitable. Herein, a multifunctional anti-scarring platform (PVA@rGO-Ag/5-Fu) integrated with outstanding photothermal, antibacterial and drug delivery abilities is developed. PVA@rGO-Ag shows favorable biocompatibility as well as an accurate regional photothermal killing ability on both conjunctival fibroblasts and bacteria under 808 nm near-infrared (NIR) irradiation. Furthermore, PVA@rGO-Ag/5-Fu improves bleb survival rates and results in the satisfactory reduction of intraocular pressure (IOP) by decreasing the fibrous reaction in vivo. In summary, PVA@rGO-Ag/5-Fu has promising potential as an efficacious and safe anti-scarring agent for filtering surgery.

Melatonin ameliorates paclitaxel-induced mice spermatogenesis and fertility defects.

Chemotherapeutic drug of paclitaxel (PTX) has been shown to cause reproductive toxicity thus affecting male fertility, but its underlying molecular basis is unclear. Melatonin (MLT) can mitigate the reproductive damage caused by certain chemotherapy drugs. In this study, we aimed to identify impact of PTX on the main biological processes and protective effect of MLT on reproductive damage caused by PTX. Mice exposed to PTX mainly impaired spermatogenesis, such as decreased sperm counts, reduced sperm motility and increased abnormal sperm. Decreased expressions of germ cell proliferation-associated protein PCNA and meiosis-related protein SYCP3 induced by PTX were determined by Western blot, while MLT ameliorated this effect and increased the expressions of PCNA, SYCP3, DMC1, STRA8 and fertility-related protein of HSPA2, resulting in significantly improved spermatogenesis and sperm quality levels. In vitro fertilization experiment showed that PTX significantly decreased blastocyst formation rates, which can be improved by MLT administration, but not two-cell development rates. Taken together, this work demonstrated PTX can adversely affect germ cell proliferation and meiosis, which ultimately influence sperm quality and male fertility, and highlighted the protective ability of MLT on ameliorating the side effects of PTX, especially on sperm quality. The results provide information to further the study on the molecular mechanism of PTX's effects on male reproduction and the protective mechanism of MLT.

Cytokinins is involved in regulation of tomato pericarp thickness and fruit size.

Although cytokinins (CKs) regulate fruit development, no direct genetic evidence supports the role of endogenous CKs in pericarp growth or development or fruit size. Here, we report that the reduction in endogenous active CKs level via overexpression of a CKs-inactivating enzyme gene AtCKX2 specifically in fruit tissues resulted in reduced pericarp thickness and smaller fruit size, compared to wild-type control fruits. The pericarp thickness and single fruit weight in transgenic plants were significantly reduced. Analysis of paraffin sections showed that the reduced pericarp thickness was due largely to a decreased number of cells, and thus decreased cell division. Transcriptome profiling showed that the expression of cell division- and expansion-related genes was reduced in AtCKX2-overexpressing fruits. In addition, the expression of auxin-signaling and gibberellin-biosynthetic genes was repressed, whereas that of gibberellin-inactivating genes was enhanced, in AtCKX2-overexpressing fruits. These results demonstrate that endogenous CKs regulate pericarp cell division and, subsequently, fruit size. They also suggest that CKs interact with auxin and gibberellins in regulating tomato pericarp thickness and fruit size.

Expressions of HSPA1L and HSPA9 are associated with poor sperm quality of low-motility spermatozoa in fertile men.

Human semen is a heterogeneous group containing a portion of low-motility sperm, which may determine the sperm quality evaluation. Abnormally expressed proteins in low-motility spermatozoa will be the candidates for sperm biology research. By comparing proteomes of high- or low-motility spermatozoa from the same semen of normal fertile men, 21 differentially expressed proteins were identified. Proteins with molecular chaperone function were significantly over-represented, of which HSPA1L and HSPA9 significantly decreased in low-motility sperm. Compared with young adult testes with normal spermatogenesis, HSPA1L and HSPA9 had decreased expressions in elderly testis characterised with poor spermatogenesis, suggesting their associations with spermatogenesis. Decreased expressions of HSPA1L and HSPA9 in low-motility spermatozoa were validated by Western Blot and immunofluorescence quantification analysis. HSPA1L was mainly expressed on sperm post-acrosome and midpiece, whilst HSAP9 was mainly expressed on acrosome and sperm tail. HSPA1L antibody could inhibit sperm motility validated by antibody blocking experiment, whilst HSPA9 antibody showed no significant effect on sperm motility. The study demonstrated that low-motility spermatozoa from fertile men had poor sperm quality, in which differential expressed proteins were promising markers for evaluating sperm quality, understanding mechanism of male infertility with unexplained causes, and providing new idea for male infertility research.

In Situ Functionalization of Silver Nanoparticles by Gallic Acid as a Colorimetric Sensor for Simple Sensitive Determination of Melamine in Milk.

A simple and green colorimetric sensing assay strategy for highly efficient determination of melamine has been fabricated, which is based on the redox reaction of gallic acid with Ag+. Monodispersed Ag nanoparticles (AgNPs) were obtained using gallic acid as a reducing and stabilizing agent. However, the aggregate behavior of AgNPs was observed, while the melamine was present in the reaction medium. As a result, the color of the solution changed from vivid yellow to brown, and the density of the color was quantitatively correlated with the melamine concentration. The aggregation of AgNPs could be attributable to the formation of hydrogen bonds between melamine and gallic acid. The designed sensor exhibited a good detection limit of 0.099 µM (0.012 ppm), which was much lower than the safety limit in China (1.0 ppm) and EU (2.0 ppm). Additionally, the sensing assay displayed good selectivity toward melamine over other coexisting substances. Consequently, the proposed colorimetric sensor was successfully used for the determination of melamine detection in raw milk samples.