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Many microorganisms have mechanisms that protect cells against attack from viruses. The fermentation components of Streptomyces sp. 1647 exhibit potent anti-influenza A virus (IAV) activity. This strain was isolated from soil in southern China in the 1970s, but the chemical nature of its antiviral substance(s) has remained unknown until now. We used an integrated multi-omics strategy to identify the antiviral agents from this streptomycete. The antibiotics and Secondary Metabolite Analysis Shell (antiSMASH) analysis of its genome sequence revealed 38 biosynthetic gene clusters (BGCs) for secondary metabolites, and the target BGCs possibly responsible for the production of antiviral components were narrowed down to three BGCs by bioactivity-guided comparative transcriptomics analysis. Through bioinformatics analysis and genetic manipulation of the regulators and a biosynthetic gene, cluster 36 was identified as the BGC responsible for the biosynthesis of the antiviral compounds. Bioactivity-based molecular networking analysis of mass spectrometric data from different recombinant strains illustrated that the antiviral compounds were a class of structural analogues. Finally, 18 pseudo-tetrapeptides with an internal ureido linkage, omicsynins A1–A6, B1–B6, and C1–C6, were identified and/or isolated from fermentation broth. Among them, 11 compounds (omicsynins A1, A2, A6, B1–B3, B5, B6, C1, C2, and C6) are new compounds. Omicsynins B1–B4 exhibited potent antiviral activity against IAV with the 50% inhibitory concentration (IC50) of approximately 1 µmol∙L–1 and a selectivity index (SI) ranging from 100 to 300. Omicsynins B1–B4 also showed significant antiviral activity against human coronavirus HCoV-229E. By integrating multi-omics data, we discovered a number of novel antiviral pseudo-tetrapeptides produced by Streptomyces sp. 1647, indicating that the secondary metabolites of microorganisms are a valuable source of novel antivirals.

Aims:Synthesis and biological evaluation of some [(5-oxazolyl)-phenyl]-thiourea derivatives as potential antiviral agents.Background:(5-Oxazolyl)-phenyl derivatives were derived from the design of mycophenolic acid structurally related analogues. The (5-oxazolyl)-phenyl fragment is an excellent composition for many novel structure compounds having good pharmaceutical properties, such as immunosuppressive, antiviral and anticancer. In the present study, we present combinations of thiourea group and (5-oxazolyl)-phenyl fragment. The antiviral activity, cytotoxicity and IMPDH activity of the title compounds were evaluated in vitro bioassay.Objective:[(5-Oxazolyl)-phenyl]-thiourea derivatives containing different substituted benzene rings were synthesized by introducing thiourea linker. All the synthesized derivatives were screened for their in vitro antiviral evaluation and inosine monophosphate dehydrogenase activity.Methods:A series of [(5-oxazolyl)-phenyl]-thiourea derivatives were synthesized by the reaction of thiocarbonyldiimidazole with amines. This was an effective method for introducing the thiourea group in the (5-oxazolyl)-phenyl structure. All of the synthesized derivatives were screened for their in vitro antiviral activity against influenza A virus, coxsackievirus B3, herpes simplex virus type 1 and inosine monophosphate dehydrogenase activity.Result:The results of the screening revealed that compounds 4i, 4j, 4k, 7m, 7n and 7o showed comparable activity towards IMPDH as compared to the control drug. Compounds 4k, 4l, 7m and 7n exhibited potent activity towards both RNA virus influenza A virus, coxsackievirus B3 and DNA virus HSV-1 at low micromolar concentrations. The activities of most compounds directly linked to the substituted benzene ring by the thiourea group were superior to those of the compounds which had the methylene linkage.Conclusion:The in vitro biological assays indicated that most of target molecules having combinations of thiourea group and (5-oxazolyl)-phenyl fragment exhibited antiviral activity and IMPDH activity compared the control drugs.

Fabrication of high-performance sensory materials for the ultra-sensitive detection of antibiotics in water is of significant importance for community health and environmental quality. Herein, two Zn(II)-based metal–organic frameworks, {[Zn3(cbbi)2(bpe)(H2O)6]·2H2O}n (FCS-4) and {[Zn4(OH)2(cbbi)2(bpee)(H2O)4]·2H2O}n (FCS-5), were prepared hydrothermally and employed as chemosensors for the ultra-sensitive detection of nitrofuran antibiotics. Interestingly, different spacers and flexibility in the nitrogen-donor ancillary ligands resulted in different topological structures and fluorescence properties. FCS-5 exhibited much better sensitivity for nitrofurazone and nitrofurantoin detection, with limits of detection (LOD) of 0.22 and 0.15 ppm, respectively. Furthermore, mechanism exploration indicated that photon-induced electron transfer plays a key role in the ultra-sensitive detection of nitrofuran antibiotics.

The development of sensitive methods for alkaline phosphatase (ALP) activity analysis is an important analytical topic. Based on the stimulus-responsive lanthanide coordination polymer, a simple ratiometric fluorescence sensing strategy was proposed to detect ALP activity. A carbon dot (CD) doped fluorescent supramolecular lanthanide coordination polymer (CDs@Tb-GMP) was prepared with Tb3+ and the ligand guanine single nucleotide (GMP). To construct a ratiometric fluorescence biosensor, the fluorescence of Tb-GMP was used as a response signal, and the fluorescence of CDs was used as a reference signal due to its good stability. When excited at 290 nm, the polymer network Tb-GMP emits characteristic fluorescence at 545 nm, while the CDs encapsulated in the polymer network emit fluorescence at 370 nm. After adding ALP to the system, the substrate GMP can be hydrolyzed by ALP, resulting in the destruction of the polymer network. Accordingly, the fluorescence of Tb-GMP significantly decreased, while the fluorescence of CDs slightly increased due to their release from the polymer network. By comparing the relationship between the fluorescence intensity ratio of the two signals and the concentration of ALP, sensitive detection of ALP could be achieved with the linear range from 0.5 to 80 U L−1 and a detection limit of 0.13 U L−1. Furthermore, the proposed ratiometric sensing system was applied to the detection of ALP in human serum samples with desirable results, indicating potential application in clinical diagnosis.

The construction of libraries of stereoisomers of natural products serves as an important approach to investigating the correlation between the stereostructure and biological activity. However, the total synthesis and isomerzation of polycyclic scaffolds with multiple chrial centers are rare. Spirooliganin (1), a new skeleton natural product isolated from the plant Illicium oligandrum, was structurally characterized by comprehensive analysis of NMR spectroscopic data and ECD which revealed an unprecedented 5–6–6–6–7 polycyclic framework with six chiral centers. Here we report a 17-step total synthesis to prepare a library of stereochemically diverse isomers of spirooliganin, including 16 diastereoisomers and 16 regioisomers. In addition to a regioselective hetero-Diels–Alder cycloaddition, the synthetic strategy involves a photo-induced stereoselective Diels–Alder reaction, which gives only the abnormal trans-fused product as rationalized by density functional theory calculations. Preliminary biological evaluation showed that spirooliganin and regioisomers 39 exhibited potent inhibition of Coxsackievirus B3. It also revealed the pharmacophore effect of the D-ring (16R,18R,24R, and 26R) for their antiviral activities., Library construction of stereochemically diverse isomers to investigate the relationship between stereoconfiguration and anti-coxsackie virus B3 activity.

Chronic graft-versus-host disease (cGVHD) remains a common threat after allogeneic hematopoietic cell transplantation (allo-HSCT), and ocular manifestations occur in up to 60% to 90% of cGVHD patients. We sought to reveal major metabolic dysregulation and to determine tear metabolites as potential biomarkers for ocular cGVHD. Twenty-three ocular cGVHD and 16 control tear samples were collected for this study. Differential metabolites were identified using a liquid chromatography-mass spectrometry system. Spearman's test was used to analyze the correlation between metabolites and ophthalmic indexes (National Institutes of Health [NIH] eye score, fluorescein tear film break-up time [T-BUT], corneal fluorescein staining [CFS], and Schirmer's test). Receiver operating characteristic (ROC) curve was analyzed to evaluate the prediction potential of identified metabolites for ocular cGVHD. Differential metabolites were mainly observed in lipid metabolites, and we highlighted the lipid dysregulation in glycerophospholipid metabolism, sphingolipid metabolism, and biosynthesis of unsaturated fatty acids. In glycerophospholipid metabolism, phosphatidylcholine (34:1) (PC [34:1]) exhibited the strongest correlation with NIH eye score (r = 0.80), T-BUT (r = 0.79), CFS (r = 0.77), and Schirmer's test (r = 0.69). In sphingolipid metabolism, sphingomyelin (SM) was the most consistent with T-BUT (r = 0.74) and CFS (r = 0.71), whereas lactosylceramide (LacCer) was the most consistent with NIH eye score (r = 0.76) and Schirmer's test (r = 0.64). In biosynthesis of unsaturated fatty acids, docosahexaenoic acid (DHA) had the highest correlation with NIH eye score (r = 0.73), T-BUT (r = 0.60), CFS (r = 0.67) and Schirmer's test (r = 0.67) (P.0001 for all). ROC analysis revealed that area under the curve (AUC) values for PC (34:1) (AUC = 0.967), LacCer (AUC = 0.946), SM (AUC = 0.932), and DHA (AUC = 0.929) were significantly correlated with cGVHD (P.0001 for all). Our study identified PC (34:1), SM, LacCer, and DHA as promising tear biomarkers to indicate metabolic dysregulation and ophthalmic manifestations in ocular cGVHD.

The human retina is a complex neural tissue that detects light and sends visual information to the brain. However, the molecular and cellular processes that underlie aging primate retina remain unclear. Here, we provide a comprehensive transcriptomic atlas based on 119,520 single cells of the foveal and peripheral retina of humans and macaques covering different ages. The molecular features of retinal cells differed between the two species, suggesting the distinct regional and species specializations of the human and macaque retinae. In addition, human retinal aging occurred in a region- and cell-type- specific manner. Aging of human retina exhibited a foveal to peripheral gradient. MYO9A− rods and a horizontal cell subtype were greatly reduced in aging retina, indicating their vulnerability to aging. Moreover, we generated a dataset showing the cell-type- and region- specific gene expression associated with 55 types of human retinal disease, which provides a foundation to understand the molecular and cellular mechanisms underlying human retinal diseases. Together, these datasets are valuable for understanding the molecular characteristics of primate retina, as well as the molecular regulation of aging progression and related diseases.

The human retina is a complex neural tissue that detects light and sends visual information to the brain. However, the molecular and cellular processes that underlie aging primate retina remain unclear. Here, we provide a comprehensive transcriptomic atlas based on 119,520 single cells of the foveal and peripheral retina of humans and macaques covering different ages. The molecular features of retinal cells differed between the two species, suggesting the distinct regional and species specializations of the human and macaque retinae. In addition, human retinal aging occurred in a region- and cell-type- specific manner. Aging of human retina exhibited a foveal to peripheral gradient. MYO9A− rods and a horizontal cell subtype were greatly reduced in aging retina, indicating their vulnerability to aging. Moreover, we generated a dataset showing the cell-type- and region- specific gene expression associated with 55 types of human retinal disease, which provides a foundation to understand the molecular and cellular mechanisms underlying human retinal diseases. Together, these datasets are valuable for understanding the molecular characteristics of primate retina, as well as the molecular regulation of aging progression and related diseases.

Colaphellus bowringi Baly mainly damages cruciferous vegetables, leading to huge economic losses. The secretory insecticidal protein (Sip) of Bacillus thuringiensis (Bt) has high insecticidal activity against C. bowringi Baly. The tertiary structure of Sip1Aa protein was analyzed by homologous modeling and other bioinformatics methods to predict the conserved domain of Sip1Aa protein. Acidic and basic amino acids in the conserved domain were selected, and alanine was used to replace these amino acids by site-directed mutation. The difference between the insecticidal activities of mutant protein and Sip1Aa protein was analyzed. The insecticidal activities of H99A, K109A, K128A, and E130A against C. bowringi Baly were significantly increased, among which that of K128A was the most obviously changed, and the LC50 value was decreased by about 10 times compared with that of Sip1Aa protein. The LC50 value of mutant E130A was 0.286 μg/mL, which was about six times less than that of Sip1Aa. K128 and E130 were both in the β9–β10 loop. The toxicity of D290A, H242A, and H303A to C. bowringi Baly was significantly reduced, and their LC50 value increased by about six, eight, and three times compared with that of Sip1Aa protein, respectively. This study showed that acidic and basic amino acid residues played a certain role in the toxicity of Sip1Aa protein, and the loss of side chains in key residues had a significant impact on the insecticidal activity of the protein. This study provides the theoretical basis for revealing the relationship between the structure and function of Sip1Aa protein and also provides a new method for the subsequent study of sip gene.

Two new metal-drug complexes constructed from non-toxic zinc and theophylline (TPL), an anti-asthmatic active drug, have been introduced into a release system based on matrices of hydroxypropylmethylcellulose (HPMC) and microcrystalline cellulose (MC). The release rate of TPL from the metal-drug complexes could be controlled by the amount of MC added, and the release mechanism changed from anomalous transport to Fickian diffusion.

Alzheimer disease (AD) is most common neurodegenerative disorder of dementia, as we all know that ApoE4 is the greatest genetic risk factor of late-onset Alzheimer's disease (LOAD). Coronary heart disease (CHD) leads to one-fourth of all deaths in industrialized countries, it is reported that ApoE4 increases the risk of coronary heart disease as well. Furthermore, evidence show that coronary heart disease also increases the incidence of Alzheimer's disease. Whether ApoE4 is a bridge connecting AD with CHD or not? And what are the special mechanism and therapeutic methods? Researchers found that cholesterol metabolic disorder is the common cause and risk factor of AD and CHD. Epidemiological studies demonstrate that carriers of the ApoE4 allele have higher cholesterol plasma concentration. More evidence indicate that hypercholesterolemia accelerates the progression of coronary atherosclerosis, damages the central nervous system blood-brain barrier, promotes Aβ protein production and Tau deposition in brain. Therefore, ApoE4 is likely to be the bridge between AD and CHD, and may be a potentially promising therapeutic target.

Porcine circovirus (PCV) is a potentially harmful virus that has been shown to contaminate biological products. The virus is resistant to many inactivation and/or removal procedures performed during manufacturing. Anion exchange chromatography has been shown to be useful for PCV type 1 (PCV1) removal; however, reduction of PCV1 using methods such as heat inactivation, low pH, nanofiltration, UV-C, and gamma irradiation has not been successful. Therefore, in this study, we evaluate various conditions for process solutions during nanofiltration using PCV1. The results indicated that PCV could be effectively removed from glycine solution at 0.1–0.3 M, pH 4.0 without IgG, using a nanofilter with a pore size of 19 nm (19-nm filter); log reduction values (LRVs) of ≥4.5 and ≥ 5.0, respectively, were obtained. In contrast, PCV1 was significantly removed (LRV: 2.2) in glycine solution at 0.3 M, pH 6.0 with 1.0% IgG using the 19-nm filter, but some virus genomes were detected in the filtrates. In summary, the use of a 19-nm filter in glycine solution with/without IgG is an appropriate condition for PCV removal.

The illegal adulteration of melamine in dairy products for false protein content increase is a strong hazard to human health. Herein, a simple and sensitive colorimetric method was developed for the quantification of melamine in dairy products based on a Cu2+-hydrogen peroxide (H2O2)-3,3′,5,5′-tetramethylbenzidine (TMB) system. In this strategy, Cu2+ exhibits peroxidase-like activity and can catalyze the oxidation of TMB to oxidized TMB (oxTMB) in the presence of H2O2 with a blue colour change of the solution. However, the presence of melamine quickly interacts with H2O2 leading to the consumption of H2O2 and thus strongly hinders the oxidation of TMB. Under the optimal conditions, the absorbance change of oxTMB has a linear response to the concentration of melamine from 1 to 100 μM with a detection limit of 0.5 μM for melamine. The proposed method has many merits including more simplicity, good selectivity, and more cost-effectiveness without using any nanomaterials. The method was further successfully applied to detect melamine in dairy products including milk and infant formula powder.

Diabetes is a chronic inflammatory disease that accelerate cardiovascular disease (CVD). However, the contribution of hyperglycemia to inflammation and CVD risk is unclear. Animal studies have shown that monocytes carry cholesterol to the atherosclerotic plaque. Thus, it is key to determine if hyperglycemia can exert a pro-atherogenic effect in human monocytes in vivo. We performed octreotide euglycemic-hyperglycemic clamps in 21 healthy subjects, 20-45 yo. Monocytes isolated during euglycemia (90 mg/dl) and after 2h of hyperglycemia (300 mg/dl) were evaluated for cholesterol metabolism and monocyte function (adhesion to fibronectin, transwell migration assays). Also, we assessed the effects of hyperglycemia on monocyte cholesterol metabolism and function from 10 healthy controls cultured in euglycemia or high glucose media with or without 2-deoxyglucose (2DG) at 2 and 6 hours. Acute hyperglycemia in vivo increased 37% total monocyte cholesterol content (TC) (p Disclosure R.M. Zhang: None. J. Oh: None. X. Lin: None. P. Stone: None. K.J. Wharton: None. A. Arbelaez: None. C. Bernal-Mizrachi: None. Funding National Heart, Lung, and Blood Institute (R01HL094818-06); National Institute of Neurological Disorders and Stroke (P01NS080675)

High-responsivity phototransistors with a structure of perovskite-WS2 nanosheet composite optical absorber and a reduced graphene oxide (rGO) channel layer is demonstrated via a facile and low-cost solution-processing method. The WS2 nanosheets are dispersed within the perovskite matrix, forming the perovskite-WS2 bulk heterojunction (BHJ). The hybrid phototransistor exhibits excellent figures of merit including high photoresponsivity of 678.8 A/W, high specific detectivity of 4.99 × 1011 Jones, high EQE value of 2.04 × 105% and rapid response to photoswitching. The high photoresponsivity could be attributed to the WS2 nanosheets induced photo-generated electron-hole separation promotion effects due to the selective electron trapping effects in the WS2 nanosheets, together with the high carrier mobility of the rGO channel. This work provides a promising platform for constructing high-responsivity photodetectors.

Background/Aims: Retinal neurodegeneration is an early event in the pathological process of diabetic retinopathy (DR). Retinal ganglion cell (RGC) injury is an important pathological feature during neurodegenerative process. Protecting RGCs from high glucose-induced injury is a promising strategy for delaying or hindering diabetes mellitus-related retinal neuropathy. This study aims to investigate the role of Nmnat1, an enzyme which catalyzes a key step in the biosynthesis of nicotinamide adenine dinucleotide (NAD), in high glucose-induced RGC injury. Methods: Western blot and immunofluorescence analysis was conducted to detect Nmnat1 expression pattern in the retina and RGC-5 cell. MTT assay, Hoechst staining, trypan blue staining, and calcein-AM/ propidium iodide (PI) staining was conducted to determine the effect of Nmnat1 knockdown on RGC-5 cell function. Microarray and bioinformatics analysis was conducted to identify potential signaling pathways affected by Nmnat1 knockdown. Pharmacological intervention, molecular intervention, and in vitro experiments were conducted to reveal molecular mechanism of Nmnat1-mediated protective effect on RGC-5 cell function. Results: Nmnat1 is constitutively expressed in retina and RGC-5 cells. Nmnat1 knockdown aggravates RGC injury, and accelerates the development of RGC-5 cell apoptosis upon high glucose stress. MAPK signaling is the primary signaling pathway affected by Nmnat1 knockdown. Under high glucose stress, Nmnat1 knockdown leads to p38-MAPK signaling inactivation. p38-MAPK pathway inhibitor strongly blocks Nmnat1-mediated protective effect on RGC-5 cell function. Conclusion: Nmnat1 protects RGC against high glucose-induced injury via p38-MAPK signaling pathway. Nmnat1 may serve as a neuroprotective target for diabetes mellitus-related retinal neuropathy.

Purpose To identify the feasibility of reconstructing corneal endothelial sheets by seeding non-infected monoclonal human corneal endothelial cells (HCECs) onto porcine Descemet's membrane (DM) and verifying the function in vitro and in vivo. Methods Denuded porcine DM was decellularized for haematoxylin and eosin staining, and DNA was removed via incubation with ethylene glycol diglycidyl ether (EGDE). The physical properties of the incubated DMs were evaluated and compared to those of unincubated DMs. The non-infected monoclonal HCECs were examined by chromosome analysis and the cell proliferation was evaluated by BrdU-labelling. Then HCECs at passage 30 were then seeded on the DM and cultured for approximately 5 days. The cell growth, density and expression of the sodium-potassium adenosine triphosphatase (Na+/K+-ATPase), the tight-junction-associated protein zonula occludens (ZO-1) and acetylated alpha tubulin were examined by electron microscopy and immunocytochemistry to compare HCECs cultivated on porcine DM and those cultured in vitro. Cells on the reconstructed HCEC sheets were labeled with DiI, and the sheets were subsequently transplanted into cat eyes via DM endothelial keratoplasty (DMEK). The corneal transparency, thickness, anterior segment, and HCEC density were monitored in vivo, and the corneal endothelial cell morphology and histological structure were examined ex vivo 98 days after surgery. Results No significant differences were observed in the elongation at break of the DMs and the thickness of the DMs incubated with EGDE compared to those of the unincubated DMs (P > 0.05). Results of chromosome analysis shown the number of the HCEC cell line was still 46 and no abnormal chromosome structure was found. BrdU-labelling shown the HCECs stopped proliferating after 5 days and the cells formed a single layer. The cells transferred to porcine DM formed tight connections with the substrate and generated layers of hexagonal cells on day 5. Adjacent cells cultivated on DM were closely attached to each other, tightly adhered to the porcine DM and expressed the Na+/K+-ATPase, ZO-1 protein and acetylated alpha tubulin, as did HCECs cultured in vitro. In addition, the HCEC density on DMs was 3020.14 ± 52.30 cells/mm2. After surgery, the corneas gradually became transparent, and the thickness decreased to 525.33 ± 56.23 μm at day 98 after the transplantation, while the control corneas showed consistent oedema during the monitoring period. The HCEC density was 2521.60 ± 78.24 cells/mm2 in vivo 98 days after transplantation. The histological results showed that the DiI-labeled cells were dense in the transplanted area and had a hexagonal or polygonal morphology and a normal ultrastructure; adjacent cells were closely attached to each other and tightly adhered to the porcine DM. Conclusions Seeding non-infected monoclonal HCECs on porcine DM could reconstruct functional corneal endothelial sheets. These results may help uncover new applications for tissue-engineered endothelium in endothelial keratoplasty.

Pathological ocular angiogenesis commonly results in visual impairment or even blindness. Unveiling the mechanisms of pathological angiogenesis is critical to identify the regulators and develop effective targeted therapies. Here, we used corneal neovascularization (CNV) model to investigate the mechanism of pathological ocular angiogenesis. We show that N6-methyladenosine (m6A) mRNA demethylation mediated by fat mass- and obesity-associated protein (FTO) could regulate endothelial cell (EC) function and pathological angiogenesis during CNV. FTO levels are increased in neovascularized corneas and ECs under pathological conditions. In vitro silencing of FTO in ECs results in reduced cellular proliferation, migration, and tube formation under both basal and pathological conditions. Furthermore, FTO silencing attenuates suture-induced CNV in vivo. Mechanically, FTO silencing in ECs could increase m6A methylation levels in critical pro-angiogenic genes, such as FAK, leading to decreased RNA stability and increased RNA decay through m6A reader YTHDF2. Our study demonstrates that FTO regulates pathological ocular angiogenesis by controlling EC function in an m6A-YTHDF2-dependent manner.

Purpose. To evaluate the safety and pharmacokinetic changes of ganciclovir (GCV) intraocular injection. Methods. GCV (2 mg/0.1 mL) was injected into rabbit eyes. Aqueous GCV concentration was detected by high-performance liquid chromatography. Potential toxicity was assessed by slit-lamp examination, optical coherence tomography, fundus examination, confocal microscopy, and histology. Results. Aqueous GCV concentrations were 24.83 ± 6.41 μg/mL, 0.65 ± 0.52 μg/mL, and undetected on the 1st, 3rd, and 7th day after intravitreal injection. GCV could not be detected on the first day after intracameral injection. No corneal abnormality was found after intravitreal injection, but retinal edema was observed on the first day which receded later. Corneal edema was obvious with endothelial cytoarchitecture damaged after intracameral injection; fluid retention also existed in retina. Conclusions. GCV intravitreal injection offers effective, sustained drug concentration in the anterior chamber, and its damage to retina receded over time. Intracameral injection results in rapid drug elimination and severe damage to endothelium and thus is not recommended.

Voltage‑gated sodium channel β2 (Navβ2), as an unconventional substrate of β‑site amyloid precursor protein cleaving enzyme 1, is involved in regulating the neuronal surface expression of sodium channels. A previous study demonstrated that knockdown of Navβ2 protected neurons and induced spatial cognition improvement by partially reducing pathological amyloidogenic processing of amyloid precursor protein (APP) in aged APP/presenilin 1 (PS1) transgenic mice. The present study aimed to investigate whether Navβ2 knockdown altered APP metabolism via regulation of the Aβ‑degrading enzyme neprilysin (NEP). APPswe/PS1ΔE9 mice (APP/PS1 transgenic mice with a C57BL/6J genetic background) carrying a Navβ2‑knockdown mutation (APP/PS1/Navβ2‑kd) or without Navβ2 knockdown (APP/PS1) were used for cell culture and further analysis. The present results demonstrated that in APP/PS1 mouse‑derived neurons, Navβ2 knockdown partially reversed the reduction in pathological APP cleavage, and the recovery of neurite extension and neuron area. Additionally, Navβ2 knockdown increased NEP activity and levels, and the levels of intracellular domain fragment binding to the NEP promoter. The present findings suggested that knockdown of Navβ2 reversed the APP/PS1 mutation‑induced deficiency in amyloid β degradation by regulating NEP.

Glaucoma is a neurodegenerative disease characterized by retinal ganglion cell (RGC) loss, optic disc excavation, and progressive visual field loss. Direct or indirect ameliorating retinal neurodegeneration is a promising therapeutic therapy for glaucoma. Circular RNAs (circRNAs) are a class of covalently closed circular RNA transcripts and have emerged as potential regulators in several neurodegenerative diseases. In this study, we show that cZRANB1 expression is significantly upregulated in retinal neurodegeneration induced by glaucoma. cZRANB1 knockdown decreases retinal reactive gliosis, glial cell activation, and facilitates RGC survival in vivo. cZRANB1 knockdown directly regulates Müller cell function and indirectly regulates RGC function in vitro. cZRANB1 acts as miRNA sponge to regulate Müller cell function through cZRANB1/miR-217/RUNX2 network. Intervention of cZRANB1 expression would become an effective strategy for treating retinal neurodegeneration.

Background/Aims: Parkinson’s disease (PD) is a frequently occurring condition that resulted from the loss of midbrain neurons, which synthesize the neurotransmitter dopamine. In this study, we established mouse models of PD to investigate the expression of microRNA-128 (miR-128) and mechanism through which it affects apoptosis of dopamine (DA) neurons and the expression of excitatory amino acid transporter 4 (EAAT4) via binding to axis inhibition protein 1 (AXIN1). Methods: Gene expression microarray analysis was performed to screen differentially expressed miRNAs that are associated with PD. The targeting relationship between miR-128 and AXIN1 was verified via a bioinformatics prediction and dual-luciferase reporter gene assay. After separation, DA neurons were subjected to a series of inhibitors, activators and shRNAs to validate the mechanisms of miR-128 in controlling of AXIN1 in PD. Positive protein expression of AXIN1 and EAAT4 in DA neurons was determined using immunocytochemistry. miR-128 expression and the mRNA and protein levels of AXIN1 and EAAT4 were evaluated via RT-qPCR and Western blot analysis, respectively. DA neuron apoptosis was evaluated using TUNEL staining. Results: We identified AXIN1 as an upregulated gene in PD based on the microarray data of GSE7621. AXIN1 was targeted and negatively mediated by miR-128. In the DA neurons, upregulated miR-128 expression or sh-AXIN1 increased the positive expression rate of EAAT4 together with mRNA and protein levels, but decreased the mRNA and protein levels of AXIN1, apoptosis rate along with the positive expression rate of AXIN1; however, the opposite trend was found in response to transfection with miR-128 inhibitors. Conclusion: Evidence from experimental models revealed that miR-128 might reduce apoptosis of DA neurons while increasing the expression of EAAT4 which might be related to the downregulation of AXIN1. Thus, miR-128 may serve as a potential target for the treatment of PD.

A new molecular mechanism is proposed to explain the shape memory effect of mesogen-jacketed liquid crystalline polymer fiber—poly-{2,5-bis[(4-butoxyphenyl)oxycarbonyl]styrene} (PBPCS) fiber-designed in the molecular level. Due to the strong “coupling effect” between flexible backbone and rigid side mesogens, the molecular structure of PBPCS can be regarded as “dual chains” model that is constituted by a flexible backbone chain and a rigid side chain in parallel. The flexible backbone chains are reversible above the transition temperature and act as a switch structure, the physically cross-linked points caused by the π–π weak interactions between rigid side mesogens are responsible for memorizing a shape function and act as a fixing structure; therefore, the switch structure (flexible backbone) and the fixing structure (rigid mesogens) are found in each macromolecular chain. Furthermore, the shape memory behavior of PBPCS fibers is discussed by a viscoelastic molecular model consisting of two Maxwell models in parallel, the obtained theoretical values by “dual chains” model was in agreement with experimental results, very well. The viscoelastic molecular model described for shape memory process was correlated to the molecular mechanism explained for shape memory effect. This work provides a new idea and significant approach for design of new shape memory polymers.

In order to eliminate the scattered light induced by the signal amplifier in fluorescence anisotropy (FA) assays, a nanosized metal–organic framework MIL-101, ranging from 80–500 nm, has been synthesized through a hydrothermal method with the addition of glycerol. We chose the 100 nm MIL-101 to enhance FA for label-free detection of the respiratory syncytial virus (RSV) gene sequence and the DNA-intercalating dye SYBR Green I (SGI) as the fluorophore, based on the different affinities of MIL-101 toward ssDNA and dsDNA. The nanosized MIL-101 has a negligible scattering effect owing to its smaller particle size, so all of the experimental data of FA values were smaller than the maximum initial anisotropy of 0.4. As a specific advantage, a dual amplification result of not only an increase in the FA value of SGI/ssDNA (r1) but also a decrease in the FA value of SGI/dsDNA (r2) was presented at the same time. Consequently, a larger FA value change Δr (Δr, Δr = r1 − r2) was obtained and contributed to improve the sensitivity. In addition, the quantitative detection of the target DNA (T) was achieved according to the relationship between Δr and the concentration of T. In the presence of MIL-101, the Δr is 7-fold higher than that without MIL-101 and achieved the sensitive and selective detection of RSV DNA.

Macrophage motility is vital in innate immunity, which contributes strategically to the defensive inflammation process. During bacterial infection, lipopolysaccharide (LPS) potently activates the migration of macrophages via the NF-κB/iNOS/c-Src signaling pathway. However, the downstream region of c-Src that participates in macrophage migration is unclear. EFhd2, a novel actin bundling protein, was evaluated for its role in LPS-stimulated macrophage migration in this study. We found that LPS stimulated the up-regulation, tyrosine phosphorylation and membrane translocation of EFhd2 in macrophages. The absence of EFhd2 inhibited the recruitment of macrophages in the lungs of LPS-induced septic mice. LPS-induced macrophage migration was neutralized by the deletion of EFhd2. EFhd2-mediated up-regulation of NFPs (including Rac1/Cdc42, N-WASP/WAVE2 and Arp2/3 complex) induced by LPS could be used to explain the role of EFhd2 in promoting actin polymerization. Furthermore, the purified EFhd2 could directly promote actin polymerization in vitro. Dasatinib, a c-Src specific inhibitor, inhibited the up-regulation of EFhd2 stimulated by LPS. Therefore, our study demonstrated that EFhd2 might be involved in LPS-stimulated macrophage migration, which provides a potential target for LPS-activated c-Src during macrophage mobilization.

Voltage-gated sodium channels beta 2 (Navβ2, encoded by SCN2B) is a substrate of β-site amyloid precursor protein cleaving enzyme 1 (BACE1) and regulates cell surface expression of channels in neurons. Previous studies reported enhanced Navβ2 processing by BACE1 in Alzheimer's disease (AD) model and patients. We investigated whether changes in Navβ2 expression affect neuronal seizure and amyloid precursor protein (APP) processing in an AD mouse model. Our study used eight-month-old APP/presenilin 1 (PS1) mice and transgenic Navβ2 knockdown [by 61% vs. wild type (WT)] APP/PS1 mice (APP/PS1/Navβ2-kd), with age-matched WT and Navβ2 knockdown (Navβ2-kd) mice as controls. We found that Navβ2 knockdown in APP/PS1 mice partially reversed the abnormal Navβ2 cleavage and the changes in intracellular and total Nav1.1α expression. It also restored sodium currents density in hippocampal neurons and neuronal activity, as indicated by EEG tracing; improved Morris water maze performance; and shifted APP amyloidogenic metabolism towards non-amyloidogenic processing. There were no differences in these indicators between WT and Navβ2-kd mice. These results suggest Navβ2 knockdown may be a promising strategy for treating AD.

Tetrathiafulvalene (TTF) and its derivatives are unique π-electron donor and they show rich redox properties. In this paper, two new conjugated C∧N ligands based on TTF ( La and Lb ) have been synthesized and their structures are determined by X-ray crystallography. Iridium complexes, [Ir( La )2(tpip)] ( 1 , tpip=tetraphenylimido-diphosphinate) and [Ir( Lb )2(tpip)] ( 2 ), based on the two ligands have been synthesized. All new compounds are characterized by elemental analysis, IR, 1H NMR and MALDI-TOF-MS methods. The electrochemical and spectroscopic properties including UV-Vis absorption spectra and photoluminescence for 1 and 2 are studied.

A coordination polymer formulated as {[Mn2L(N3)4]·2H2O}n(1) [L = 1,4-bis(pyridinil-3-carboxylato)-l,4-dimethylbenzene] was synthesized and structurally and magnetically characterized. The uniform Mn(II) chains with mixed (μ-EO-N3)2(μ-COO) triple bridges (EO = end-on) are linked by L ligands to generate a 2-fold interpenetrating 3D framework. Meanwhile, magnetism analysis reveals antiferromagnetic coupling for 1.

Two series of lanthanide metal-organic frameworks, [KLn3(FDCA)5(H2O)5]·solvent (1Ln) (Ln = La 1, Ce 2, Pr 3), [K4Ln8(FDCA)14(H2O)30]·2(H2FDCA)·DMAC·10(H2O) (2Ln) (Ln = Eu 4, Gd 5, Tb 6, Dy 7, Er 8, DMAC = N,N-dimethylacetamide), have been prepared by solvothermal reactions of 2,5-furandicarboxylic acid (H2FDCA) and KNO3 with the corresponding lanthanide nitrate. X-ray crystal structure analyses show that 1Ln possesses a 3D network based on the “16-crown-6-like” secondary building unit (SBU), whereas 2Ln exhibits a 1D framework with a “23-crown-9-like” SBU. The structural investigation unambiguously demonstrated that the lanthanide contraction affects the formation of 1Ln and 2Ln. Meanwhile, the solid-state photoluminescent properties of 4 and 6 were also investigated at room temperature and the luminescence intensity of 4 is sensitive to small molecules.

A series of novel unsaturated five-membered benzo-heterocyclic amine derivatives were synthesized and assayed to determine their in vitro broad-spectrum antiviral activities. The biological results showed that most of our synthesized compounds exhibited potent broad-spectrum antiviral activity. Notably, compounds 3f (IC50 = 3.21–5.06 μM) and 3g (IC50 = 0.71–34.87 μM) showed potent activity towards both RNA viruses (influenza A, HCV and Cox B3 virus) and a DNA virus (HBV) at low micromolar concentrations. An SAR study showed that electron-withdrawing substituents located on the aromatic or heteroaromatic ring favored antiviral activity towards RNA viruses.

A series of novel (5-oxazolyl)phenyl amine derivatives were synthesized and their antiviral activities against the hepatitis C virus (HCV) and the coxsackie virus B3 (CVB3) and B6 (CVB6) were evaluated in vitro. Bioassays showed that the synthesized compounds 17a1, 17a4, 17a6, 17b1, 17d1, 17e2 and 17g3 exhibited potent antiviral activity against HCV (IC₅₀ = 0.28-0.92 μM) and most synthesized compounds exhibited low cytotoxicity in Huh7.5 cells, compared to telaprevir. The compounds 17a1, 17a4, 17a5, 17a6, 17b1, 17b2, 17g1 and 17g3 showed strong activity against the CVB3 and/or CVB6 at low concentrations (IC₅₀ < 2.0 μM). The (5-oxazolyl)phenyl amines 17a1, 17a4, 17a8, 17b1, 17d1, 17e2, 17f3 and 17g3 were identified as the most active on the biological assays, and will be studied further.

New dinuclear complexes with the bridging S―S coupled and dimerized quinoxaline-2,3-dithiolate (dsqdt) ligand, [TpPh2M(dsqdt)MTpPh2] (TpPh2 = hydro-tris(3,5-diphenylpyrazol-1-yl)borate; M = Co, 1; M = Ni, 2; M = Mn, 3), were prepared. X-ray crystal structure studies for complexes 1–3 indicate that the dsqdt ligand forms the unique κ2-coordinated mode through S and N atoms from the same qdt unit. The absorption spectra, redox behavior and magnetic properties of the compounds are reported.

Two novel spirooliganones A (1) and (2), a pair of spiro carbon epimers, with a rare dioxaspiro skeleton were isolated from the roots of Illicium oligandrum. The structures were fully determined by spectroscopic analysis and chemical methods, especially modified Mosher's method, and X-ray diffraction analysis. Spirooliganone B was found to exhibit more potent activities against coxsackie virus B3 and influenza virus A (H3N2) (IC50 3.70-5.05 μM) than spirooliganone A. The biosynthetic pathway involving a hetero-Diels-Alder reaction of the epimers was proposed.

A new tetrathiafulvalene (TTF) derivative possessing Schiff base core with the coumarin moiety has been synthesized and characterized. The crystal structure has been studied, and the electrochemical and spectroscopic properties have also been investigated. Optimized conformation and molecular orbital diagram of the title compound have been calculated with density functional theory (DFT).

A series of novel N-phenylbenzamide derivatives were synthesized and their anti-EV 71 activities were assayed in vitro. Among the compounds tested, 3-amino-N-(4-bromophenyl)-4-methoxybenzamide (1e) was active against the EV 71 strains tested at low micromolar concentrations, with IC50 values ranging from 5.7 ± 0.8–12 ± 1.2 μM, and its cytotoxicity to Vero cells (TC50 = 620 ± 0.0 μM) was far lower than that of pirodavir (TC50 = 31 ± 2.2 μM). Based on these results, compound 1e is a promising lead compound for the development of anti-EV 71 drugs.

Background/Aims: Podocyte injury, especially podocyte apoptosis, plays a major role in early-stage diabetic nephropathy (DN). Swiprosin-1, also known as EF hand domain containing 2 (EFhd2), is a Ca2+-binding protein in different cell types. However, the function of swiprosin-1 in podocytes remains unknown. Methods: The expression and distribution of swiprosin-1 were investigated in the mouse renal glomerulus and conditionally immortalized mouse podocyte cell line MPC-5. The expression of swiprosin-1 was also detected in streptozotocin (STZ)-treated mice and MPC-5 cells treated with high glucose (HG). Nephrin and podocin were detected by immunohistochemistry and immunofluroscence. Collagen IV, transforming growth factor-β (TGF-β) and fibronectin mRNA expressions were assayed by real-time PCR. Apoptotic proteins and phosphorylation of p38 mitogen-activated protein kinase (MAPK) were detected by immunoblotting. Results: Swiprosin-1 was found to be expressed in podocytes of the mouse glomerulus and MPC-5 cells. Swiprosin-1 expression was increased in STZ-treated mice and MPC-5 cells treated with HG. In Swiprosin-1-/- diabetic mice, kidney/ body weight, urinary albumin, podocyte foot process effacement and glomerular basement membrane thickening were attenuated; the downregulation of nephrin and podocin expression in the glomerulus was inhibited; and the upregulation of collagen IV, TGF-β and fibronectin mRNA expression in the renal cortex was ameliorated as compared with those in diabetic swiprosin-1+/+ mice. In addition, the increased apoptosis of podocytes, proapoptotic protein expression and p38 phosphorylation in Swiprosin-1-/- diabetic mice were inhibited as compared with those in diabetic swiprosin-1+/+ mice. Knockdown of swiprosin-1 in MPC-5 cells reduced the apoptosis of podocytes, proapoptotic protein expression and p38 phosphorylation induced by HG. Targeted knockdown of p38 attenuated the increased apoptosis of MPC-5 cells over-expressing swiprosin-1. Conclusion: Swiprosin-1 expression in podocytes of the mouse glomerulus played a critical role in early-stage DN. Swiprosin-1 deficiency in early DN attenuated mitochondria-dependent podocyte apoptosis induced by hyperglycemia or HG via p38 MAPK signaling pathway.

To observe the effect of puerarin on learning and memory function and tau phosphorylation in APP/PS1 transgenic mice, drugs were administered to 3-month old APP/PS1 transgenic mice. Learning and memory function of mice were assessed by Morris water maze test 3 months after treatment. Animals were decapitated after behavioral test. The levels of Aβ were detected by ELISA, the expression of protein [tau, phosphorylated tau, GSK3β and p-GSK3β(Ser9)] were assessed by Western blot. Morris water maze test showed that the escape latency of APP/PS1 double transgenic mice was significantly longer than that of the normal control group, and the residence time of the original quadrant was significantly shorter. The escape latency of puerarin group was significantly shorter and the residence time of the original quadrant was prolonged compared with the model group. Compared with the normal control group, the levels of Aβ in the cortex of APP/PS1 transgenic mice were increased, the expression of phosphorylated tau was significantly increased, and the expression of phosphorylated GSK3β(Ser9) protein was decreased. Treatment with puerarin, the latency of APP/PS1 transgenic mice was significantly reduced, the level of Aβ was decreased, the expression of phosphorylated tau was significantly decreased, and the expression of phosphorylated GSK3β(Ser9) protein was increased. Puerarin improves the learning and memory impairment by reducing the formation of Aβ, activating the GSK3β signaling pathway, inhibiting the phosphorylation of tau in APP/PS1 double transgenic mice.

A novel class of thioflavone and flavonoid derivatives has been prepared and their antiviral activities against enterovirus 71 (EV71) and the coxsackievirus B3 (CVB3) and B6 (CVB6) were evaluated. Compounds 7d and 9b showed potent antiviral activities against EV71 with IC 50 values of 8.27 and 5.48 μM, respectively. Compound 7f , which has been synthesized for the first time in this work, showed the highest level of inhibitory activity against both CVB3 and CVB6 with an IC 50 value of 0.62 and 0.87 μM. Compounds 4b , 7a , 9c and 9e also showed strong inhibitory activities against both the CVB3 and CVB6 at low concentrations (IC 50 =1.42−7.15 μM), whereas compounds 4d , 7c , 7e and 7g showed strong activity against CVB6 (IC 50 =2.91–3.77 μM) together with low levels of activity against CVB3. Compound 7d exhibited stronger inhibitory activity against CVB3 (IC 50 =6.44 μM) than CVB6 (IC 50 >8.29 μM). The thioflavone derivatives 7a , 7c , 7d , 7e , 7f and 7g , represent a new class of lead compounds for the development of novel antiviral agents.

New π-extended tetrathiafulvalenes (exTTF) 4a, 4b, 5a, and 5b have been synthesized by direct phosphite-mediated cross-couplings of anthraquinone with 1,3-dithiole-2-thione derivatives. Further coordination reactions of the exTTF ligands including the phenanthroline unit (5a and 5b) with [Re(CO)5Cl] afford interesting tricarbonylrhenium(I) complexes 6a and 6b, respectively. The X-ray crystal structures of compounds 4a and 4b show a saddlelike conformation of the exTTF framework. The electrochemical and spectroscopic properties of compounds 4a–6a and 4b–6b have been studied. The results suggest that the oxidation of the exTTF derivatives shows the typical quasi-reversible two-electron oxidation wave of the exTTF core at potentials that vary depending on the substituents.

Based on the developed low permeability test instrument, gas transport properties of typical Jin-Ping marbles are studied under steady flow of nitrogen. A low permeability test instrument and its tri-axial cell structure were introduced in detail, and this test instrument was used to study transport properties of dense rocks under different temperatures and stress conditions, the Test results showed that intrinsic permeability of Jin-Ping marble is about 10－20 m2. Comparisons are made between test results and an exact method which considers Klinkenberg effect in gas flow equation. Fitting results using the exact method show better agreement with laboratory testing results and the transport parameters gained are more convincible.

Metal ion sensors are an important yet challenging field in analytical chemistry. Despite much effort, only a limited number of metal ion sensors are available for practical use because sensor design is often a trial-and-error-dependent process. DNAzyme-based sensors, in contrast, can be developed through a systematic selection that is generalizable for a wide range of metal ions. Here, we summarize recent progress in the design of DNAzyme-based fluorescent, colorimetric, and electrochemical sensors for metal ions, such as Pb2+, Cu2+, Hg2+, and UO22+. In addition, we also describe metal ion sensors based on related DNA molecules, including T-T or C-C mismatches and G-quadruplexes.

FeS-1, VS-1 or CuS-1 precursors are loaded and highly dispersed on the surface of mesoporous materials SBA-15, MCM-41 or SiO2by impregnating method. The samples were characterized by X-ray diffraction, FT-IR spectroscopy, TEM and N2adsorption. Their catalytic performances for styrene epoxidation with anhydrous TBHP as an oxidant agent were studied. The results suggest that zeolite primary and secondary building units are highly dispersed on the samples and the samples have still retained mesoporous structure. All of the catalysts exhibit good catalytic activities, producing styrene oxide and benzaldehyde as the main products. Cu/SBA-15 shows high activity and selectivity, which are comparable with that of other samples. We find that, under the conditions of preferable precursors,n(TBHP)/n(styrene) =1.5, reaction temperature 80°C, and reaction time 2.0h, the conversion and yield of styrene epoxide reached 65.9% and 42.2%, respectively.

Candida infections have become an increasingly significant problem, mainly because of the widespread nature of Candida and drug resistance. There is an urgent need to develop new classes of drugs for the treatment of opportunistic Candida infections, especially in medically complex patients. Previous studies have confirmed that 2-amino-nonyl-6-methoxyl-tetralin muriate (10b) possesses powerful antifungal activity in vitro against Candia albicans. To clarify the underlying action mechanism, an oligonucleotide microarray study was performed in C. albicans SC5314 without and with 10b treatment. The analytical results showed that energy metabolism-related genes, including glycolysis-related genes (PFK1, CDC19 and HXK2), fermentation-related genes (PDC11, ALD5 and ADH1) and respiratory electron transport chain-related genes (CBP3, COR1 and QCR8), were downregulated significantly. Functional analysis revealed that 10b treatment increased the generation of endogenous reactive oxygen species, and decreased mitochondrial membrane potential, ubiquinone–cytochrome c reductase (complex III) activity and intracellular ATP levels in C. albicans SC5314. Also, addition of the antioxidant ascorbic acid reduced the antifungal activity of 10b significantly. These results suggest that mitochondrial aerobic respiration shift and endogenous reactive oxygen species augmentation might contribute to the antifungal activity of 10b against C. albicans. This information may prove to be useful for the development of new strategies to treat Candida infections.

Net ecosystem exchange (NEE) of carbon dioxide (CO2) was measured at Zoige wetland using the eddy covariance technique. Analysis of CO2 fluxes in two years showed Zoige wetland was a net CO2 sink of −47.1 and −79.7 g C m−2 a−1 in 2008 and 2009, respectively. The peak NEE value was −0.54 mg CO2 m−2 s−1 (the negative value signifies net ecosystem carbon gain from air). The maximal daily integrated NEE was −4.1 g C m−2 d−1 during the peak growth season (from July to August). Gross ecosystem photosynthesis was likely more variable than ecosystem respiration at both seasonal and interannual timescales in this wetland. Our data strongly suggested that the combination of precipitation and temperature, as well as phenological stage of vegetation, controlled the dynamics of ecosystem carbon gain, even in drought years. Therefore, an accurate representation of these parameters in climate models is critical to the success of forecasting carbon budgets of alpine wetlands.

The objective was to evaluate the effect of various vitrification conditions on the morphology of bovine secondary and primordial follicles, and to use xenografting to confirm their developmental ability. Secondary follicles were placed in vitrification solution containing 15% (v:v) ethylene glycol (EG), 15% (v:v) dimethyl sulfoxide (DMSO), 20% (v:v) fetal calf serum (FCS), and 0, 0.25, or 0.5 M sucrose at room temperature for 1 or 30 min, or at 4 degrees C for 30 min before being plunged into liquid nitrogen (LN(2)). Ovarian tissues with primordial follicles were equilibrated in a solution containing 7.5% EG, 7.5% DMSO, and 20% FCS for 5 or 15 min, and then treated with a vitrification solution (15% EG, 15% DMSO, and 20% FCS) containing 0 or 0.5 M sucrose at room temperature for 1 min, and then plunged into LN(2). One week later, follicles and tissues were warmed, and morphology assessed histologically. Secondary follicles vitrified in sucrose-free solution had more oocytes with shrinkage of the nucleus and abnormal cytoplasm relative to those vitrified in sucrose-containing solution. When primordial follicles were equilibrated for 5 min and vitrified in sucrose-free solution, the percentage of morphologically normal primordial follicles was higher than in the other groups (P < 0.05). After 4 wk and 6 mo of xenografting of vitrified-warmed secondary and primordial follicles, respectively, in SCID mice, follicles developed to the antral stage and oocytes grew. In conclusion, bovine secondary follicles were successfully cryopreserved in sucrose-containing vitrification solutions and maintained their ability to develop to the antral stage and grow oocytes, whereas primordial follicles vitrified in sucrose-free solution maintained their morphology and developed to the antral stage, with oocyte growth.

To identify changes in the gene expression profile of Candida albicans (C albicans) biofilms following exposed to 2-amino-nonyl-6-methoxyl-tetralin muriate(10b) and clarify the mechanism of 10b against C albicans biofilms.Anti-biofilm activity of 10b was assessed by tetrazolium (XTT) reduction assay and the action mechanism against biofilms was investigated by cDNA microarray analysis and real-time RT-PCR assay.Ten differentially expressed genes were directly linked to biofilm formation and filamentous or hyphal growth (eg, NRG1, ECE1 and CSA1). Decreased gene expression was involved in glycolysis (eg, HXK2 and PFK1) and antioxidant defense (eg, SOD5), while increased gene expression was associated with enzymes that specifically hydrolyzed beta-1,3 glucan (XOG1), and with lipid, fatty acid and sterol metabolism (eg, SLD1, ERG6 and ERG2). Functional analysis indicated that addition of anti-oxidant ascorbic acid reduced inhibitory efficiency of 10b on mature biofilm.Inhibition of 10b on biofilm formation possibly depends on impairing the ability of C albicans to change its morphology via altering the expression of biofilm formation genes. Mitochondrial aerobic respiration shift and endogenous ROS augmentation might be a major contribution to reduce mature biofilm metabolic activity. The data may be useful for the development of new strategies to reduce the incidence of device-associated infections.