An enhanced cascade-based deep forest model for drug combination prediction.

Combination therapy has shown an obvious curative effect on complex diseases, whereas the search space of drug combinations is too large to be validated experimentally even with high-throughput screens. With the increase of the number of drugs, artificial intelligence techniques, especially machine learning methods, have become applicable for the discovery of synergistic drug combinations to significantly reduce the experimental workload. In this study, in order to predict novel synergistic drug combinations in various cancer cell lines, the cell line-specific drug-induced gene expression profile (GP) is added as a new feature type to capture the cellular response of drugs and reveal the biological mechanism of synergistic effect. Then, an enhanced cascade-based deep forest regressor (EC-DFR) is innovatively presented to apply the new small-scale drug combination dataset involving chemical, physical and biological (GP) properties of drugs and cells. Verified by the dataset, EC-DFR outperforms two state-of-the-art deep neural network-based methods and several advanced classical machine learning algorithms. Biological experimental validation performed subsequently on a set of previously untested drug combinations further confirms the performance of EC-DFR. What is more prominent is that EC-DFR can distinguish the most important features, making it more interpretable. By evaluating the contribution of each feature type, GP feature contributes 82.40%, showing the cellular responses of drugs may play crucial roles in synergism prediction. The analysis based on the top contributing genes in GP further demonstrates some potential relationships between the transcriptomic levels of key genes under drug regulation and the synergism of drug combinations.

Relative corneal refractive power shift and inter-eye differential axial growth in children with myopic anisometropia treated with bilateral orthokeratology.

PURPOSE: To investigate the relationship between relative corneal refractive power shift (RCRPS) and axial length growth (ALG) in bilateral myopic anisometropes treated with orthokeratology. METHODS: A total of 102 children with myopic anisometropia in this prospective interventional study were randomly assigned to the spectacle group and orthokeratology group. Axial length (AL) and corneal topography was measured at baseline and the 12-month follow-up visit. ALG was defined as the difference between the two measurements, and RCRPS profiles were calculated from two axial maps obtained. RESULTS: In the orthokeratology group, the ALG in the more myopic eye (0.06 ± 0.15 mm) was significantly smaller than that in the less myopic eye (0.15 ± 0.15 mm, p < 0.001), and the interocular difference in AL significantly decreased following 1-year treatment, from 0.47 ± 0.32 to 0.38 ± 0.28 mm (p < 0.001). However, in the spectacle group, the ALG was similar between the two eyes, and the interocular difference in AL did not change significantly over one year (all p > 0.05). The interocular difference in ALG in the orthokeratology group was significantly correlated with the interocular difference in RCRPS (dRCRPS, ß=-0.003, p < 0.001) and the interocular difference in baseline AL (ß=-0.1179, p < 0.001), with R2 being 0.6197. CONCLUSION: Orthokeratology was effective in decreasing the magnitude of anisometropia. The interocular variation in RCRPS is an important factor accounting for the reduction of interocular ALG difference in anisomyopic children post-orthokeratology. These results provide insight into establishing eye-specific myopia control guidelines during orthokeratology treatment for myopic anisometropes.

Integrated omic profiling of the medicinal mushroom Inonotus obliquus under submerged conditions.

BACKGROUND: The Inonotus obliquus mushroom, a wondrous fungus boasting edible and medicinal qualities, has been widely used as a folk medicine and shown to have many potential pharmacological secondary metabolites. The purpose of this study was to supply a global landscape of genome-based integrated omic analysis of the fungus under lab-growth conditions. RESULTS: This study presented a genome with high accuracy and completeness using the Pacbio Sequel II third-generation sequencing method. The de novo assembled fungal genome was 36.13 Mb, and contained 8352 predicted protein-coding genes, of which 365 carbohydrate-active enzyme (CAZyme)-coding genes and 19 biosynthetic gene clusters (BCGs) for secondary metabolites were identified. Comparative transcriptomic and proteomic analysis revealed a global view of differential metabolic change between seed and fermentation culture, and demonstrated positive correlations between transcription and expression levels of 157 differentially expressed genes involved in the metabolism of amino acids, fatty acids, secondary metabolites, antioxidant and immune responses. Facilitated by the widely targeted metabolomic approach, a total of 307 secondary substances were identified and quantified, with a significant increase in the production of antioxidant polyphenols. CONCLUSION: This study provided the comprehensive analysis of the fungus Inonotus obliquus, and supplied fundamental information for further screening of promising target metabolites and exploring the link between the genome and metabolites.

The relationship between baseline axial length and axial elongation in myopic children undergoing orthokeratology.

PURPOSE: To investigate the correlation between the baseline axial length (AL) and axial elongation in myopes undergoing orthokeratology (ortho-k). METHODS: This was a retrospective study. During the 1-year follow-up, 1176 children (aged 8-14 years) were included and divided into an ortho-k group (n = 588) and a single-vision spectacle group (n = 588). The ortho-k group participants (8-11 years of age) who completed the 3-year follow-up (n = 150) were further divided into three subgroups stratified by their baseline AL: subgroup 1 (AL < 24.5 mm), subgroup 2 (24.5 ≤ AL < 26 mm) and subgroup 3 (AL ≥ 26 mm). AL was measured at baseline and during the annual visit. RESULTS: The ortho-k group exhibited slower 1-year axial elongation (39% reduction) than the spectacle group. The 1-year axial elongation was negatively correlated with initial age in both groups. A negative association between 1-year axial elongation and baseline AL was observed in the ortho-k group but not in the spectacle group. However, this relationship only existed in ortho-k participants 8-11 years of age. For the younger ortho-k participants who completed the 3-year follow-up, the annual axial elongation was significantly higher in subgroup 1 for the first and second years but not in the third year compared with subgroups 2 and 3. CONCLUSION: Axial elongation was negatively correlated with baseline AL in the ortho-k group. Children aged 8-11 years with longer baseline AL (≥24.5 mm) demonstrated slower annual axial elongation during the first 2 years of ortho-k treatment, which may provide insight into establishing individual guidelines for controlling myopia using ortho-k in children with different baseline characteristics.

Cranial Bone Transport Promotes Angiogenesis, Neurogenesis, and Modulates Meningeal Lymphatic Function in Middle Cerebral Artery Occlusion Rats.

BACKGROUND: Ischemic stroke is a leading cause of death and disability worldwide. However, the time window for quickly dissolving clots and restoring cerebral blood flow, using tissue-type plasminogen activator treatment is rather limited, resulting in many patients experiencing long-term functional impairments if not death. This study aims to determine the roles of cranial bone transport (CBT), a novel, effective, and simple surgical technique, in the recovery of ischemic stroke using middle cerebral artery occlusion (MCAO) rat model. METHODS: CBT was performed by slowly sliding a bone segment in skull with a special frame and a speed of 0.25 mm/12 hours for 10 days following MCAO. Morris water maze, rotarod test, and catwalk gait analysis were used to study the neurological behaviors, and infarct area and cerebral flow were evaluated during CBT process. Immunofluorescence staining of CD31 and Nestin/Sox2 (sex determining region Y box 2) was performed to study the angiogenesis and neurogenesis. OVA-A647 (ovalbumin-Alexa Fluor 647) was intracisterna magna injected to evaluate the meningeal lymphatic drainage function. RESULTS: CBT treatment has significantly reduced the ischemic lesions areas and improved the neurological deficits in MCAO rats compared with the rats in the control groups. CBT treatment significantly promoted angiogenesis and neurogenesis in the brain of MCAO rats. The drainage function of meningeal lymphatic vessels in MCAO rats was significantly impaired compared with normal rats. Ablation of meningeal lymphatic drainage led to increased neuroinflammation and aggravated neurological deficits and ischemic injury in MCAO rats. CBT treatment significantly improved the meningeal lymphatic drainage function and alleviated T-cell infiltration in MCAO rats. CONCLUSIONS: This study provided evidence for the possible mechanisms on how CBT attenuates ischemic stroke injury and facilitates rapid neuronal function recovery, suggesting that CBT may be an alternative treatment strategy for managing ischemic stroke.

Alleviation of osteoarthritis by intra-articular transplantation of circulating mesenchymal stem cells.

This study aimed to evaluate the efficacy of intra-articular delivery of peripheral blood derived mesenchymal stromal cells (PB-MSCs) on the progression of trauma-induced osteoarthritis (OA) in mice. Adult male C57BL/6J mice subjected to destabilization of the medial meniscus surgeries (DMM) were randomly divided into four groups: sham surgery group; vehicle control group (treated with saline), PBMSC-treated group, or adipose tissue derived MSCs (AD-MSC)-treated group (n = 4 per group). PB-MSCs and AD-MSCs were harvested and cultured following previously established protocols, and pre-labeled with BrdU for 48 h before transplantation. PB-MSCs or AD-MSCs (5 × 105 cells/mouse; passage 3-5) were intra-articular injected into the right knee joints thrice post-surgery. The mice were euthanized at 8 weeks post-surgery and knee joint samples were collected for micro-CT and histological examinations. PB-MSCs administration significantly reduced subchondral bone volume comparing to the vehicle control group. Safranin O staining showed that PB-MSCs treatment ameliorated degeneration of articular cartilage, which was comparable to AD-MSCs treatment. The expression of catabolic marker MMP13 was significantly reduced in articular cartilage of the PB-MSCs treated group comparing to that of the vehicle control group. Co-expression of BrdU and Sox9 indicated that injected PB-MSCs differentiated in chondrocytes in situ, along with reduced levels of IL-6 within peripheral sera of PB-MSCs- and AD-MSCs-treated mice. Therefore, administration of PB-MSCs or AD-MSCs attenuated trauma-induced OA progression through inhibiting cartilage degradation and inflammation. PB-MSCs are ideal cell source for treating cartilage-associated diseases.

Comprehensive identification of protein orthologs in the family Ascoviridae facilitates an understanding of phylogenomics, protein conservation, and phosphorylation.

Analysis of orthology is important for understanding protein conservation, function, and phylogenomics. In this study, we performed a comprehensive analysis of gene orthology in the family Ascoviridae based on identification of 366 protein homologue groups and phylogenetic analysis of 34 non-single-copy proteins. Our findings revealed 90 newly annotated proteins, five newly identified core proteins for the family Ascoviridae, and 14 core proteins for the genus Ascovirus. A phylogenomic tree of 11 Ascoviridae members was constructed based on a concatenation of 35 of the 45 ortholog groups. In combination with phosphoproteomic results and conservation estimations, 30 conserved phosphorylation sites on 17 phosphoproteins were identified from a total of 176 phosphosites on 57 phosphoproteins from Heliothis virescens ascovirus 3h (HvAV-3h), providing potential research targets for investigating the role of these protein in the regulation of viral infection. This study will facilitate genome annotation and comparison of further Ascoviridae members as well as functional genomic investigations.

The treatment zone decentration and corneal refractive profile changes in children undergoing orthokeratology treatment.

BACKGROUND: To confirm the association between treatment-zone (TZ) decentration and axial length growth (ALG) in children who underwent orthokeratology; and to explore the association between TZ decentration and relative corneal refractive power (RCRP) profile, which was known to be significantly associated with ALG retardation. METHODS: Four hundred myopic children of age 12 years participated in the study, with 200 wearing orthokeratology lenses and the other 200 wearing single-vision spectacle as the controls. Cycloplegic refraction was performed at baseline. Axial length was measured at baseline and 12 months after initial lens wear, and ALG was defined as the difference. In the ortho-k group, TZ decentration and the RCRP map were calculated from the topography map obtained at the 12-month visit. RCRP were summed within various chord radii from the cornea center, and the association to TZ decentration, spherical equivalent (SE), ALG were analyzed with linear regressions. RESULTS: Compared to the controls, children wearing orthokeratology lenses had significantly smaller ALG over 1 year (0.1 ± 0.15 mm vs. 0.32 ± 0.17 mm, p < 0.001). ALG was significantly and negatively associated with summed RCRP within the central cornea of 2 mm in radius. The mean TZ decentration was 0.62 ± 0.25 mm, and the mean direction was 214.26 ± 7.39 degrees. ALG was negatively associated with the TZ decentration magnitude (p < 0.01), but not the direction (p = 0.905). TZ decentration caused an asymmetrical distribution of the RCRP with the nasal side plus power shifting towards the corneal center. For chord radius ranging 1-2 mm, the association between TZ decentration and the summed RCRP were significant, and the proportion of variance accountable increased with chord radius. For chord radius beyond 1.5 mm, the association between baseline spherical equivalent (SE) and summed RCRP was significant. The portion of variance accountable by SE increased and peaked in 2.5 mm chord radius. CONCLUSIONS: A larger TZ decentration was associated with a larger summed RCRP in the central cornea. It may be one of the possible reasons why TZ decentration is beneficial to retarding myopia progression.

Vasoactive Intestinal Peptide Promotes Fracture Healing in Sympathectomized Mice.

Vasoactive intestinal peptide (VIP) as a neuromodulator and neurotransmitter played a significant role in modulating bone homeostasis. Our previous study reported an essential role of VIP in in vitro BMSCs osteogenesis and in vivo bone defect repair. VIP was also revealed to have a promoting effect on embryonic skeletal element development. However, the role of VIP in fracture healing is not known yet. We hypothesized that the disorder of sympathetic nervous system impairs bone structure and fracture healing, whereas VIP may rescue the sympathetic inhibition effects and promote fracture healing. We employed a 6-hydroxydopamine (6-OHDA) induced sympathectomy mice model (sympathectomized mice), in which successful sympathetic inhibition was confirmed by a decreased level of norephedrine (NE) in the spleen. In the sympathectomized mice, the femoral micro-architecture, bone density and mechanical properties were all impaired compared to the vehicle control mice. The femoral fracture was created in the vehicle or sympathectomized mice. Vehicle mice were locally injected with PBS as a negative control, and the sympathectomized mice were treated with injection of PBS or VIP. VIP expression at the fracture site was significantly decreased in sympathectomized mice. The fracture healing was repressed upon 6-OHDA treatment and rescued by VIP treatment. Micro-CT examination showed that the femoral bone micro-architecture at the fracture sites and mechanical properties were all impaired. Simultaneously, the expression level of osteogenic markers OCN and OPN were reduced in sympathectomized mice compared with vehicle group. While the VIP treatment rescued the repression effects of 6-OHDA on bone remodeling and significantly promoted bone quality and mechanical properties as well as increased osteogenesis marker expression in the sympathectomized mice. VIP administration promoted bone fracture healing by inhibiting bone resorption, making it a putative new alternative treatment strategy for fracture healing.

The treatment zone size and its decentration influence axial elongation in children with orthokeratology treatment.

BACKGROUND: To investigate whether the treatment zone size (TZS) and treatment zone decentration (TZD) will affect the axial elongation in myopic children undergoing orthokeratology treatment. METHODS: A self-controlled retrospective study was conducted on 352 children who met the inclusion criteria. Axial length was measured before and at 12 months after the initial lens wear. Corneal topography was measured at baseline and at each follow-up after lens wear. The Corneal topography obtained from the 12-month visit was used to quantify TZS and TZD for each subject. Cycloplegic refraction was required for all children before fitting the orthokeratology lenses. RESULTS: Axial elongation was significantly associated with age, baseline spherical equivalent (SE), TZS, and TZD with univariate linear regression. In groups with both small and large TZS, axial elongation was significantly decreased with large TZD (both P < 0.01). In groups with both small and large TZD, axial elongation was significantly decreased with small TZS (P = 0.03 for small TZD, P = 0.01 for large TZD). Age, SE, and TZD were significantly associated with axial elongation in multiple regression (all P < 0.01). CONCLUSION: Relatively smaller TZS and larger TZD may be beneficial in slowing myopia progression in children with orthokeratology treatment.

Formation mechanism of the nanostructure in laser streaming phenomenon.

Laser streaming is a phenomenon in which liquid streaming is driven directly from the laser through an in situ fabricated nanostructure. In this study, liquid streaming of a gold nanoparticle suspension driven by a pulsed laser was studied using a high-speed camera. The laser streaming formation time, streaming velocity, and relative energy conversion efficiency of laser streaming was measured for different nanoparticle concentrations, focal lens position, laser powers, and laser repetition rates. In addition to the laser intensity, which played a significant role in the formation process of laser streaming, the optical gradient force was found to be an important approach involved in the transport and provision of nanoparticles during the formation of laser streaming. This finding facilitated a better understanding of the formation mechanism of laser streaming and demonstrated the possibilities of a new potential laser etching technique based on nanosecond lasers and nanoparticle suspensions. This result can also expand the application of laser streaming in microfluids and other fields that require lasers to move macroscopic objects at relatively high speeds.

Lgr5-overexpressing mesenchymal stem cells augment fracture healing through regulation of Wnt/ERK signaling pathways and mitochondrial dynamics.

Fracture remains one of the most common traumatic conditions in orthopedic surgery. The use of mesenchymal stem cells (MSCs) to augment fracture repair is promising. Leucine-rich repeat-containing GPCR 5 (Lgr5), a transmembrane protein, has been identified as a novel adult stem cell marker in various organs and tissues. However, the roles of Lgr5 in MSCs are not fully understood. In this study, we investigated cellular functions of Lgr5 in MSCs and its potential implications in treating fracture. Lgr5-overexpressing MSCs (MSCLgr5) were established in murine SV40 promoter-driven luciferase reporter MSC line through virus transfection. Results of real-time quantitative PCR and Western blot analysis confirmed the increased expression of Lgr5 in MSCLgr5. MSCLgr5 exhibited increased osteogenic capacity, which may result from elevated expression of ß-catenin and phosphorylated ERK1/2 within the nuclear region of cells. In contrast, inhibition of Lgr5 expression decreased the osteogenic differentiation ability of MSCs, accompanied with increased mitochondrial fragmentation and reduced expression of ß-catenin. Local transplantation of MSCLgr5 at fracture sites accelerated fracture healing via enhanced osteogenesis and angiogenesis. MSCLgr5 stimulated the tube formation capacity of HUVECs in a Matrigel coculture system in vitro significantly. Taken together, results suggest that Lgr5 is implicated in the cellular processes of osteogenic differentiation of MSCs through regulation of Wnt and ERK signaling pathways and mitochondrial dynamics in fusion and fission. Inhibition of Lgr5 expression induced increased mitochondrial fragmentation and suppression of osteogenesis. MSCLgr5 exhibited enhanced therapeutic efficacy for fracture healing, which may serve as a superior cell source for bone tissue repair.-Lin, W., Xu, L., Pan, Q., Lin, S., Feng, L., Wang, B., Chen, S., Li, Y., Wang, H., Li, Y., Wang, Y., Lee, W. Y. W., Sun, D., Li, G. Lgr5-overexpressing mesenchymal stem cells augment fracture healing through regulation of Wnt/ERK signaling pathways and mitochondrial dynamics.

Sox11-modified mesenchymal stem cells accelerate cartilage defect repair in SD rats.

Cartilage has a limited capacity to heal. Previously, we have shown that overexpression of Sox11 in rMSCs (Rat Mesenchymal Stem Cells) by lentivirus-mediated gene transfer leads to enhanced tri-lineage differentiation and accelerated bone formation in fracture model of rats. We observed that the fracture repair in the rats that received Sox11-modified rMSCs injection proceeded through an endochondral ossification process much faster than those in the control groups. However, the detailed role of Sox11 in rMSCs chondrogenic differentiation, as well as cartilage defect, is still not clearly clarified. Therefore, this study tests the hypothesis that Sox11 promotes chondrogenesis and cartilage defect repair by regulating ß-catenin. Sox11 was transduced into rMSCs using lentiviruses. The expression levels of ß-catenin and its downstream genes were evaluated by quantitative RT-PCR. The transcriptional activation of ß-catenin was proved by dual-luciferase reporter assay and co-immunoprecipitation was performed to evaluate Sox11-ß-catenin interaction. In addition, a cartilage defect model in SD rats was used to evaluate the cartilage regeneration ability of Sox11-modified rMSCs in vivo. We found that Sox11 transcriptionally activated ß-catenin expression and discovered the core promoter region (from - 242 to - 1414) of ß-catenin gene for Sox11 binding. In addition, Sox11 might regulate ß-catenin at the post-transcriptional level by protein-protein interaction. Finally, using a cartilage defect model in rats, we found Sox11-modified rMSCs could improve cartilage regeneration. Taken together, our study shows that Sox11 is an important regulator of chondrogenesis and Sox11-modified rMSCs may have clinical implication for accelerating cartilage defect healing.

Influence of Corneal Topographic Parameters in the Decentration of Orthokeratology.

OBJECTIVE: To investigate the lens decentration (LD) of orthokeratology (ortho-k) and the association between pretreatment corneal topographic parameters and LD of the ortho-k. METHODS: Fifty right eyes of 50 myopes wearing ortho-k lenses were included in the prospective study. Corneal topography was conducted pretreatment to get topographic corneal parameters, including flat-K (K1); steep-K (K2); corneal astigmatism (CA), CA at 0 to 3 mm (3 mm-CA), 3 to 5 mm (5 mm-CA), 5 to 7 mm (7 mm-CA); surface asymmetry index (SAI); surface regularity index; the curvature of best-fit sphere; the diameter of cornea (DC); the distance from the corneal center to the corneal vertex (CCCV); flat eccentricity (E1), steep eccentricity (E2), and E1/E2 (E ratio); and the corneal curvature differences between the nasal and temporal quadrants at 0 to 3 mm (3 mm-Knt), and the corneal curvature differences between the superior and inferior quadrants at 0 to 3 mm (3 mm-Ksi), 5 mm-Knt (at 3-5 mm), 5 mm-Ksi (at 3-5 mm), 7 mm-Knt (at 5-7 mm), and 7 mm-Ksi (at 5-7 mm). The relationship between these cornea topographic parameters and LD of the ortho-k was tested using stepwise multiple linear regression models. RESULTS: The mean magnitude of LD was 0.51±0.23 mm (0.06-1.03 mm). According to the stepwise analysis, 4 factors were associated with the overall LD (P<0.01): SAI (ß=0.252), CCCV (ß=0.539), 5 mm-CA (ß=-0.268), and 3 mm-Ksi (ß=-0.374); 5 factors were associated with the horizontal LD (P<0.01): DC (ß=0.205), CCCV (ß=0.881), 3 mm-CA (ß=-0.217), 5 mm-Knt (ß=0.15), and 3 mm-Ksi (ß=-0.18); and 3 factors were associated with the vertical LD (P<0.01): SAI (ß=0.542), 5 mm-CA (ß=-0.188), and 3 mm-Ksi (ß=-0.213). CONCLUSION: Lens decentration is most common, but in most cases, the amount of LD is moderate and acceptable. The magnitude of LD can be predetermined by topographic corneal parameters. Surface asymmetry index, CCCV, 5 mm-Knt, and 3 mm-Ksi may be more preferable parameters in terms of the assessment of LD of ortho-k.

MicroRNA-218 Promotes Osteogenic Differentiation of Mesenchymal Stem Cells and Accelerates Bone Fracture Healing.

As a regulator of osteogenesis, microRNA-218 (miR-218) is reported to promote osteogenesis of mesenchymal stem cells (MSCs). However, the in vivo osteogenic effect of miR-218 remains elusive. In this study, miR-218 was confirmed to promote osteogenic differentiation of MSCs by stimulating the alkaline phosphatase activity, calcium nodule formation, and osteogenic marker gene expression. For in vivo study, the miR-218-overexpressing BMSCs were locally administrated into the fracture sites in a femur fracture mouse model. Based on the X-rays, micro-computed tomography, mechanical testing, histology, and immunohistochemistry examinations, miR-218 overexpression improved new bone formation and accelerated fracture healing. These findings suggest that miR-218 may be a promising therapeutic target for bone repair in future clinical applications.

Efficacy of Trial Fitting and Software Fitting for Orthokeratology Lens: One-Year Follow-Up Study.

OBJECTIVE: To observe and compare the clinical efficacy of 1-year trial fitting and software fitting orthokeratology lenses. METHODS: One hundred myopes who received vision correction with the use of orthokeratology lenses form July 2016 to September 2017 were included in this study. Subjects were assigned randomly into the two groups: the trial fitting group (group A) and the software fitting group (group B). For the right eye of each subject, measurements, such as uncorrected visual acuity (UCVA, logarithm of minimal angle of resolution), refractive error, corneal topography, ocular health status, and the fitting situation, were obtained at baseline, 1 week, 1 month, 3 months, 6 months, and 12 months after lens wear. Axial length and corneal endothelium cells (CECs) were also measured at baseline and 12 months after wearing the lens. RESULTS: Compared with the baseline, the spherical equivalent refraction, UCVA, and central corneal curvature changed significantly after orthokeratology (OK) lens wear (all P<0.05). Between groups A and B, the parameters aforementioned were insignificant at each time point (all P>0.05). Axial length and CECs showed no significant changes during the first year of OK treatment (all P>0.05). Rate of corneal staining between two groups revealed no difference during 1-year visit (P<0.05). CONCLUSION: Both the trial lens fitting and software fitting approaches were effective in temporarily reducing myopia, providing good UCVA and delaying the elongation of axial length for moderate and high myopic adolescents. Both the two approaches can be combined in OK lens fitting.

Enhanced production of total flavones from Inonotus baumii by multiple strategies.

As one kind of important secondary metabolites produced by Inonotus baumii, flavones can be applied in food, medicine, and other industries due to their biological activities such as antioxidant, anticancer, and antibacterial activity. To enhance total flavone production in submerged fermentation of I. baumii, three different strategies, optimization of fermentation parameters by statistical designs including Plackett-Burman design and response surface methodology, addition of precursors and elicitors, and two-phase culture, were used. The production of total flavones (PTF) reached 1532.83 mg/L when the optimized medium was used. All precursors and elicitors can increase the PTF. The maximum PTF (2184.06 mg/L, up to 1.57-fold) was obtained with the addition of both AgNO3 and glutathione in fermentation media. Interestingly, when 0.5% (w/v) DM130 macroporous resin as adsorbent was added to fermentation broth on day 4 of culture, the highest production reached 2407.79 mg/L with this two-phase culture strategy. These methods can be further applied to large-scale industrial production and broaden the application of flavones.

Purification and characterization of a novel cell-penetrating carrier similar to cholera toxin chimeric protein.

Developing a recombinant vector for noninvasively delivering biological macromolecules into the brain is important. This study constructed and purified a protein complex based on the cholera toxin (CT) molecular structure. Enhanced green fluorescent protein (EGFP)-modified A2 subunits of CT (CTA2) were used as tracer molecules for introduction of transactivator of transcription (TAT) through the A subunit into cells. The protein complex EGFP-CTA2-TAT/(CTB)5 (CTB: B subunit of CT) was obtained using an in vitro recombination method and verified by monosialoganglioside-enzyme-linked immunosorbent assay and high performance liquid chromatography assay. The protein complexes bound more strongly to monosialoganglioside (GM1) than (CTB)5 at low concentrations (0.625-1.25 µg/mL). In vitro assays revealed that the transmembrane function of TAT was also maintained. The GM1-binding activity and cell membrane-penetrating ability suggested that a CT structure-based protein complexes could be used to design a delivery carrier for intranasal administration through GM1 binding. The expression vector introduced in this study provides a feasible expression frame for constructing several new macromolecular protein drugs for effective cell penetration.

[Identification and bactericidal activity of a novel Cathelicidin family member from skin of Bufu bufo gargarizans].

The skin transcriptome of Bufu bufo gargarizans was determined by conventional methods. A novel full length cDNA coding for a Cathelicidin precursor was identified by transcriptomic data assembling, annotation and blast search of corresponding data banks. According to the known processing methods of Cathelicidin family members, present reported novel Cathelicidin precursor of B. bufo gargarizans might be cleaved at 2 possible sites of the same precursor and generate both BG-CATH25 and BG-CATH29 as mature molecules. The deduced BG-CATH25 and BG-CATH29 were synthesized with purity>95% to evaluate the properties and bactericidal activities. The secondary structural characteristics of both BG-CATH25 and BG-CATH29 in different solutions were determined by Circular Dichroism (CD) Analysis. CD results indicated that random coil conformation were the main structural elements for both BG-CATH25 and BG-CATH29 in different buffer systems. Antimicrobial activities against tested bacterial strains were carried out by plating method. Both BG-CATH25 and BG-CATH29 showed strong antibacterial activities against Aeromonas hydrophila, with MIC values of 1.25, 10 mg•L⁻¹, respectively. However, both of them showed weak bactericidal activities against human pathogenic bacteria, like Escherichia coli (ATCC25922）,Staphylococcus aureus (ATCC25923）and Pseudomonas aeruginosa (ATCC 27853).

Role of Bax/Bcl-2 family members in green tea polyphenol induced necroptosis of p53-deficient Hep3B cells.

Green tea polyphenol (GTP) is one of the most promising chemopreventive agent for cancer; it can inhibit cancer cell proliferation and induce apoptosis through p53-dependent cell signaling pathways. Unfortunately, many tumor cells lack the functional p53, and little is known about the effect of GTP on the p53-deficient/mutant cancer cells. To understand the p53-independent mechanisms in GTP-treated p53-dificient/mutant cancer cells, we have now examined GTP-induced cytotoxicity in human hepatoma Hep3B cells (p53-deficient). The results showed that GTP could induce Bax and Bak activation, cytochrome c release, caspase activation, and necroptosis of Hep3B cells. Bax and Bak, two key molecules of mitochondrial permeability transition pore (MPTP), were interdependently activated by GTP, with translocation and homo-oligomerization on the mitochondria. Bax and Bak induce cytochrome c release. Importantly, cytochrome c release and necroptosis were diminished in Hep3B cells (Bax(-/-)) and Hep3B cells (Bak(-/-)). Furthermore, overexpression of Bcl-2 could ameliorate GTP-induced cytochrome c release and necroptosis. Together, the findings suggested that GTP-induced necroptosis was modulated by the p53-independent pathway, which was related to the translocation of Bax and Bak to mitochondria, release of cytochrome c, and activation of caspases.