Network pharmacology-based exploration identified the antiviral efficacy of Quercetin isolated from mulberry leaves against enterovirus 71 via the NF-κB signaling pathway.

Introduction: Mulberry leaf (ML) is known for its antibacterial and anti-inflammatory properties, historically documented in "Shen Nong's Materia Medica". This study aimed to investigate the effects of ML on enterovirus 71 (EV71) using network pharmacology, molecular docking, and in vitro experiments. Methods: We successfully pinpointed shared targets between mulberry leaves (ML) and the EV71 virus by leveraging online databases. Our investigation delved into the interaction among these identified targets, leading to the identification of pivotal components within ML that possess potent anti-EV71 properties. The ability of these components to bind to the targets was verified by molecular docking. Moreover, bioinformatics predictions were used to identify the signaling pathways involved. Finally, the mechanism behind its anti-EV71 action was confirmed through in vitro experiments. Results: Our investigation uncovered 25 active components in ML that targeted 231 specific genes. Of these genes, 29 correlated with the targets of EV71. Quercetin, a major ingredient in ML, was associated with 25 of these genes. According to the molecular docking results, Quercetin has a high binding affinity to the targets of ML and EV71. According to the KEGG pathway analysis, the antiviral effect of Quercetin against EV71 was found to be closely related to the NF-κB signaling pathway. The results of immunofluorescence and Western blotting showed that Quercetin significantly reduced the expression levels of VP1, TNF-α, and IL-1ß in EV71-infected human rhabdomyosarcoma cells. The phosphorylation level of NF-κB p65 was reduced, and the activation of NF-κB signaling pathway was suppressed by Quercetin. Furthermore, our results showed that Quercetin downregulated the expression of JNK, ERK, and p38 and their phosphorylation levels due to EV71 infection. Conclusion: With these findings in mind, we can conclude that inhibiting the NF-κB signaling pathway is a critical mechanism through which Quercetin exerts its anti-EV71 effectiveness.

Diagnosis of in vivo vertical root fracture using deep learning on cone-beam CT images.

OBJECTIVES: Evaluating the diagnostic efficiency of deep learning models to diagnose vertical root fracture in vivo on cone-beam CT (CBCT) images. MATERIALS AND METHODS: The CBCT images of 276 teeth (138 VRF teeth and 138 non-VRF teeth) were enrolled and analyzed retrospectively. The diagnostic results of these teeth were confirmed by two chief radiologists. There were two experimental groups: auto-selection group and manual selection group. A total of 552 regions of interest of teeth were cropped in manual selection group and 1118 regions of interest of teeth were cropped in auto-selection group. Three deep learning networks (ResNet50, VGG19 and DenseNet169) were used for diagnosis (3:1 for training and testing). The diagnostic efficiencies (accuracy, sensitivity, specificity, and area under the curve (AUC)) of three networks were calculated in two experiment groups. Meanwhile, 552 teeth images in manual selection group were diagnosed by a radiologist. The diagnostic efficiencies of the three deep learning network models in two experiment groups and the radiologist were calculated. RESULTS: In manual selection group, ResNet50 presented highest accuracy and sensitivity for diagnosing VRF teeth. The accuracy, sensitivity, specificity and AUC was 97.8%, 97.0%, 98.5%, and 0.99, the radiologist presented accuracy, sensitivity, and specificity as 95.3%, 96.4 and 94.2%. In auto-selection group, ResNet50 presented highest accuracy and sensitivity for diagnosing VRF teeth, the accuracy, sensitivity, specificity and AUC was 91.4%, 92.1%, 90.7% and 0.96. CONCLUSION: In manual selection group, ResNet50 presented higher diagnostic efficiency in diagnosis of in vivo VRF teeth than VGG19, DensenNet169 and radiologist with 2 years of experience. In auto-selection group, Resnet50 also presented higher diagnostic efficiency in diagnosis of in vivo VRF teeth than VGG19 and DensenNet169. This makes it a promising auxiliary diagnostic technique to screen for VRF teeth.

Using deep learning to distinguish malignant from benign parotid tumors on plain computed tomography images.

Objectives: Evaluating the diagnostic efficiency of deep-learning models to distinguish malignant from benign parotid tumors on plain computed tomography (CT) images. Materials and methods: The CT images of 283 patients with parotid tumors were enrolled and analyzed retrospectively. Of them, 150 were benign and 133 were malignant according to pathology results. A total of 917 regions of interest of parotid tumors were cropped (456 benign and 461 malignant). Three deep-learning networks (ResNet50, VGG16_bn, and DenseNet169) were used for diagnosis (approximately 3:1 for training and testing). The diagnostic efficiencies (accuracy, sensitivity, specificity, and area under the curve [AUC]) of three networks were calculated and compared based on the 917 images. To simulate the process of human diagnosis, a voting model was developed at the end of the networks and the 283 tumors were classified as benign or malignant. Meanwhile, 917 tumor images were classified by two radiologists (A and B) and original CT images were classified by radiologist B. The diagnostic efficiencies of the three deep-learning network models (after voting) and the two radiologists were calculated. Results: For the 917 CT images, ResNet50 presented high accuracy and sensitivity for diagnosing malignant parotid tumors; the accuracy, sensitivity, specificity, and AUC were 90.8%, 91.3%, 90.4%, and 0.96, respectively. For the 283 tumors, the accuracy, sensitivity, and specificity of ResNet50 (after voting) were 92.3%, 93.5% and 91.2%, respectively. Conclusion: ResNet50 presented high sensitivity in distinguishing malignant from benign parotid tumors on plain CT images; this made it a promising auxiliary diagnostic method to screen malignant parotid tumors.

Enterovirus 71 induces autophagy in mice via mTOR inhibition and ERK pathway activation.

AIMS: Enterovirus 71 (EV71) is one of the main viruses that cause hand-foot-mouth disease; however, its pathogenic mechanism remains unclear. This study characterized the relationship between EV71 infection and autophagy in vivo and explored the molecular mechanism underlying EV71-induced autophagy. MATERIALS AND METHODS: A mouse model of EV71 infection was prepared by intraperitoneally injecting one-day-old BALB/c suckling mice with 30 µL/g of EV71 virus stock solution for 3 days. The behavior, fur condition, weight, and mice mortality were monitored, and disease scores were calculated. The pathological damage to the brain, lung, and muscle tissues after the viral infection was assessed by hematoxylin and eosin staining. Western blot and immunofluorescence analyses were used to detect the expression levels of viral protein 1, Beclin-1, microtubule-associated protein light chain 3B, mammalian target of rapamycin (mTOR), phosphorylated (p)-mTOR, extracellular signal-regulated protein kinase (ERK) 1/2, and p-ERK. KEY FINDINGS: EV71 infection can trigger autophagy in the brains, lungs, and muscles of infected mice. The autophagy response triggered by EV71 is achieved by the simultaneous mTOR inhibition and the ERK pathway activation. Blocking the mTOR pathway may aggravate autophagy, whereas ERK inhibition alleviates autophagy but cannot completely prevent it. SIGNIFICANCE: EV71 infection can induce autophagy in mice, involving mTOR and ERK signaling pathways. These two signaling pathways are independent and do not interfere with each other.

Hydroalcoholic extract from Abelmoschus manihot (Linn.) Medicus flower reverses sleep deprivation-evoked learning and memory deficit.

Previous researches have indicated that sleep plays a vital role in cognitive functions. Sleep deprivation (SD) causes learning and memory damage, which is associated with oxidative stress. This study was performed to investigate the neuroprotective effects of an extract of Abelmoschus manihot flower (EAM) against memory deficit induced by SD in mice. The SD model was evoked by multiple platform method for 5 days, successively. The learning and memory-improving effects of EAM were assessed by behavioral trials and the underlying mechanism was investigated by measuring the oxidative stress alteration. Our findings indicated that the SD-induced memory deficit and the EAM treatment improved the cognitive functions of mice in the object location recognition test and passive avoidance task. In addition, EAM effectively improved the activities of the antioxidant enzyme, decreased the content of malondialdehyde (MDA), and restored the protein expression of the brain-derived neurotrophic factor (BDNF), tyrosine kinase B (TrkB) and glutamate receptor 1 (GluR1) in brain tissues. In conclusion, EAM could improve the SD-evoked learning and memory impairments. The possible underlying mechanisms of EAM may be related to its antioxidant capacity and enhanced BDNF/TrkB/GluR1 levels in the hippocampal memory.

CXCL3 overexpression promotes the tumorigenic potential of uterine cervical cancer cells via the MAPK/ERK pathway.

CXCL3 belongs to the CXC-type chemokine family and is known to play a multifaceted role in various human malignancies. While its clinical significance and mechanisms of action in uterine cervical cancer (UCC) remain unclear. This investigation demonstrated that the UCC cell line HeLa expressed CXCL3, and strong expression of CXCL3 was detected in UCC tissues relative to nontumor tissues. In addition, CXCL3 expression was strongly correlated with CXCL5 expression in UCC tissues. In vitro, HeLa cells overexpressing CXCL3, HeLa cells treated with exogenous CXCL3 or treated with conditioned medium from WPMY cells overexpressing CXCL3, exhibited enhanced proliferation and migration activities. In agreement with these findings, CXCL3 overexpression was also associated with the generation of HeLa cell tumor xenografts in athymic nude mice. Subsequent mechanistic studies demonstrated that CXCL3 overexpressing influenced the expression of extracellular signal-regulated kinase (ERK) signaling pathway associated genes, including ERK1/2, Bcl-2, and Bax, whereas the CXCL3-induced proliferation and migration effects were attenuated by exogenous administration of the ERK1/2 blocker PD98059. The data of the current investigation support that CXCL3 appears to hold promise as a potential tumor marker and interference target for UCC.

Characterization of a Novel Tectivirus Phage Toil and Its Potential as an Agent for Biolipid Extraction.

The oleaginous bacterium Rhodococcus opacus PD630 is metabolically diverse and can be cultivated on various renewable resources to serve as a sustainable triacylglycerol (TAG) feedstock for biodiesel production. Current methods for TAG extraction are costly, but infection of cultures by lytic bacteriophages (phages) may be a viable approach for achieving release of intracellular lipid from oleaginous bacteria such as R. opacus. This study reports the novel tectiviral phage Toil capable of releasing intracellular contents including a fluorescent protein marker and TAGs into the supernatant after phage infection of R. opacus PD631, a domesticated derivative of strain PD630. Phage Toil is placed in the Tectiviridae by its morphology, the presence of a lipid membrane, its genome architecture and the presence of terminal covalently-linked proteins. Toil is the first tectivirus capable of infecting a member of the Actinobacteria. Microscopy shows that infected cells do not undergo sudden lysis but instead maintain their original shape for several hours, with the cellular morphology gradually deteriorating. Approximately 30% of intracellular TAGs could be recovered from the culture supernatants of Toil-infected PD631 cells. Phage Toil has potential to be used as an agent in extraction of TAGs from oleaginous bacterium R. opacus. IMPORTANCE: This study reported the first tectivirus (Phage Toil) capable of infecting a member of the Actinobacteria. In this study, we showed that Phage Toil can infect oleaginous bacterium Rhodococcus opacus to release intracellular contents such as a fluorescent protein marker and TAG lipid granules, which can serve as a starting material for biodiesel production. This study demonstrates a new method to extract TAGs by using this phage. Additionally, Phage Toil can be a new model phage to advance knowledge regarding phage infection mechanisms in Rhodococcus and other mycolic acid-containing bacteria such as Mycobacterium.

Draft Genome Sequence of Staphylococcus succinus subsp. succinus Type Strain DSM 14617, Isolated from Plant and Soil Inclusions within 25- to 35-Million-Year-Old Dominican Amber.

Staphylococcus succinus subsp. succinus type strain DSM 14617 was isolated from plant and soil inclusions within 25- to 35-million-year-old Dominican amber. The complete genome sequence of strain DSM 14617T includes a genome of 2.88 Mb (32.94% G+C content) without any plasmids.

CXCL5 as an autocrine or paracrine cytokine is associated with proliferation and migration of hepatoblastoma HepG2 cells.

C-X-C motif chemokine ligand 5 (CXCL5) is a CXC-type chemokine that is a crucial inflammatory mediator and a powerful attractant for granulocytic immune cells. Increasing evidence has indicated that CXCL5 is involved in the tumorigenesis of various malignancies. The present investigation demonstrated that CXCL5 was expressed in both hepatoblastoma HepG2 cells and liver stellate LX-2 cells, and CXCL5's receptor C-X-C chemokine receptor type 2 (CXCR2) was expressed in HepG2 cells by reverse transcription-polymerase chain reaction (RT-PCR), western blotting and ELISA assays. Cell counting kit-8, colony formation and Transwell assays revealed that exogenous CXCL5 expression efficiently promoted proliferation, colony formation and migration of HepG2 cells. To explore the autocrine and paracrine roles of CXCL5 in the oncogenic potential of HepG2 cells, HepG2 cells overexpressing CXCL5 and LX-2 cells overexpressing CXCL5 were successfully constructed by gene transfection. Similarly, overexpression of CXCL5 in HepG2 also enhanced proliferation, colony formation and migration of HepG2 cells. Furthermore, the condition medium of LX-2 cells overexpressing CXCL5 affected the proliferation and migration of HepG2 cells. RT-PCR and western blotting assays were also conducted to explore whether overexpression of CXCL5 in HepG2 modulated the expression of genes. The results revealed that overexpression of CXCL5 regulated the expression of several genes, including N-myc downregulated gene 3,w B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X protein, P53, vascular endothelial growth factor, interleukin (IL)-18, IL-1ß and cystathionine-γ-lyase. In conclusion, the present findings indicate that CXCL5/CXCR2 axis contributes to the oncogenic potential of hepatoblastoma via autocrine or paracrine pathways by regulating expression of genes associated with the progression of carcinoma.

Cometabolic biodegradation of 1,2,3-trichloropropane by propane-oxidizing bacteria.

1,2,3-Trichloropropane (TCP) is an emerging groundwater pollutant and suspected human carcinogen. TCP, a recalcitrant contaminant, has been detected in the subsurface near TCP manufacture facilities and many superfund sites. Considering the toxicity and the occurence of TCP, there is a need to seek for cost-effective treatment technologies for TCP-contaminated sites. This paper investigated TCP biodegradation by propane-oxidizing bacteria (PrOB) which are known to express propane monooxygenase (PrMO). PrMO can cometabolically degrade many different contaminants. Four PrOB, Rhodococus jostii RHA1, Mycobacterium vaccae JOB5, Rhodococcus rubber ENV425 and one isolate Sphingopyxis sp. AX-A were examined for their ability to degrade TCP. All the four PrOB resting cells were able to degrade TCP. Strain JOB5 exhibited the best TCP degradation ability (vinitial = 9.7 ± 0.7 µg TCP (mg protein)-1h-1). No TCP was degraded in the presence of acetylene (an inhibitor for PrMO), suggesting that PrMO might be responsible for TCP degradation. Furthermore, competitive inhibition was observed between propane and TCP, and between trichloroethylene (TCE) and TCP.

Effect of Fanbaicao (Herba Potentillae Discoloris) oil on the expression of p21 and CDK4 in HepG2 cells.

OBJECTIVE: To research the anti-cancer mechanism of the Traditional Chinese Medicine Fanbaicao (Herba Potentillae Discoloris) oil in the human hepatoma cell line HepG2. METHODS: Gas chromatography was used to analyze the components of Fanbaicao (Herba Potentillae Discoloris). We tested the inhibitory effect of Fanbaicao (Herba Potentillae Discoloris) oil on the human hepatoma cell line HepG2 in vitro using 3-(4, 5-Dimet hylt hiazol-2-yl)-2,5-dip henyltetrazolium bromide assays. Fluorescence activating cell sorter analysis was used to examine the levels of apoptosis, and western blot and immunofluorescence were used to detect the expression of p21, p-p21 and CDK4 proteins. RESULTS: Fanbaicao (Herba Potentillae Discoloris) oil contains 45 ingredients, and L-ascorbic acid 2, 6-bispalmitate was the main component and accounted for 44.96% of total drive-off peak area. Other components included (Z)-14-met hyl-8-exadecenal- acetal (8.56%), phytol (7.74%) and lauric acid (6.31% ). Fanbaicao (Herba Potentillae Discoloris) oil treatment reduced the proliferation of HepG2 cells and the half growth inhibition concentration (IC50) was 2.03 mg/mL. Furthermore, we also observed significantly increased HepG2 cell apoptosis in a dose-dependent manner (P < 0.05). Fanbaicao (Herba Potentillae Discoloris) oil significantly increased the expression of p21 and p-p21 and significantly decreased the expression of CDK4 in HepG2 cells compared with controls (P < 0.01). CONCLUSION: Our results showed that Fanbaicao (Herba Potentillae Discoloris) oil has anti-cancer activities in HepG2 cells, which is probably related to the upregulation of p21 and p-p21 and downregulation of CDK4 expression.

[Expressions of HO-2 and CO in the corpus cavernosum of castrated rats].

OBJECTIVE: To explore the expressions of HO-2 and CO in the corpus cavernosum of castrated rats in order to further study the pathogenesis of erectile dysfunction (ED). METHODS: We randomly divided 72 male SD rats into four groups: normal control, sham operation, castration, and castration + ZnPP. We detected intracavernous pressure (ICP) and penile erection in the basic condition and after apomorphine (APO) induction, determined the expression of the HO-2 protein in the corpus cavernosum by laser scanning confocal microscopy, and measured the level of CO by spectrophotometry during different periods of penile erection. RESULTS: The ICP in the basic condition and that after APO induction and the rate of penile erection were decreased significantly in the castration group ([11.68 ± 0.69] mmHg, [54.81 ± 3.86] mmHg, and 33.3%) and the castration + ZnPP group ([11.20 ± 0.71] mmHg, [41.17 ± 5.41] mmHg, and 22.2%) as compared with the normal control ([22.83 ± 2.66] mmHg, [66.92 ± 7.77] mm-Hg, and 100%) and the sham operation group ([23.35 ±2.22] mmHg, [70.43 ?7. 22] mmHg, and 100%) (all P <0. 01). After APO induction, ICP in the castration + ZnPP group was remarkably reduced in comparison with that in the castration group (P < 0.01), and so was the expression of the HO-2 protein before and during penile erection in the castration (445.4 ± 23.7 and 847.4 ± 35.0) and the castration + ZnPP group (390.1 ± 29.7 and 526.0 ± 52.5) compared with the normal control (512.7 ±57.4 and 1145.2 ± 89.8) and the sham operation group (583.7 ± 8.0 and 1016.3 ± 79.8), the expression of the HO-2 protein significantly decreased in the castration group (445.4 ± 23.7 and 847.4 ± 35.0) (P < 0.05 or 0.01), markedly lower in the castration + ZnPP than in the castration group during penile erection (P < 0.01) but with no significant differences among the four groups after it. Before, during and after penile erection, the levels of CO were remarkably decreased in the castration ([20.59 ± 1.01], [32.53 ± 1.26], and [18.71 ± 1.22] x 10(-7) nmol/L) and the castration +ZnPP group ([12.52 ± 1.05], [21.90 ± 1.02], and [16.56 ± 0.55] x 10(-7) nmol/L) as compared with the normal control ([26.76 ± 1.41], [48.25 ± 1.01], and [27.10 ± 1.58 ] x 10(-7) nmol/L) and the sham operation group ([25.41 ± 2.09], [ 47.90 ± 1.22], and [25.67 ± 1.20] x 10(-7) nmol/L) (P < 0.05 or 0.01), significantly lower in the castration + ZnPP than in the castration group during penile erection (P < 0.01). CONCLUSION: Decreased expressions of HO-2 and CO may correlate with erectile dysfunction in castrated rats.

Biodegradation of 1,4-dioxane: effects of enzyme inducers and trichloroethylene.

1,4-Dioxane is a groundwater contaminant and probable human carcinogen. In this study, two well-studied degradative bacteria Mycobacterium vaccae JOB5 and Rhodococcus jostii RHA1 were examined for their 1,4-dioxane degradation ability in the presence and absence of its co-contaminant, trichloroethylene (TCE), under different oxygenase-expression conditions. These two strains were precultured with R2A broth (complex nutrient medium) before supplementation with propane or 1-butanol to induce the expression of different oxygenases. Both propane- and 1-butanol-induced JOB5 and RHA1 were able to degrade 1,4-dioxane, TCE, and mixtures of 1,4-dioxane/TCE. Complete degradation of 1,4-dioxane/TCE mixture was observed only in propane-induced strain JOB5. Inhibition was observed between 1,4-dioxane and TCE for all cells. Furthermore, product toxicity caused incomplete degradation of 1,4-dioxane by 1-butanol-induced JOB5. In general, the more TCE degraded, the greater extent of product toxicity cells experienced; however, susceptibility to product toxicity was found to be both strain- and inducer-dependent. The findings of this study provide fundamental basis for developing an effective in-situ remediation method for 1,4-dioxane-contaminated ground water and the first known study of 1,4-dioxane degradation by wild-type strain RHA1.

Cultivation of lipid-producing bacteria with lignocellulosic biomass: effects of inhibitory compounds of lignocellulosic hydrolysates.

Lignocellulosic biomass has been recognized as a promising feedstock for the fermentative production of biofuel. However, the pretreatment of lignocellulose generates a number of by-products, such as furfural, 5-hydroxylmethyl furfural (5-HMF), vanillin, vanillic acids and trans-p-coumaric acid (TPCA), which are known to inhibit microbial growth. This research explores the ability of Rhodococcus opacus PD630 to use lignocellulosic biomass for production of triacylglycerols (TAGs), a common lipid raw material for biodiesel production. This study reports that R. opacus PD630 can grow well in R2A broth in the presence of these model inhibitory compounds while accumulating TAGs. Furthermore, strain PD630 can use TPCA, vanillic acid, and vanillin as carbon sources, but can only use TPCA and vanillic acid for TAG accumulation. Strain PD630 can also grow rapidly on the hydrolysates of corn stover, sorghum, and grass to accumulate TAGs, suggesting that strain PD630 is well-suited for bacterial lipid production from lignocellulosic biomass.

[Effects of ganoderma lucidum spores on cytochrome C and mitochondrial calcium in the testis of NIDDM rats].

OBJECTIVE: To observe the effects of Ganoderma lucidum spores on Cytochrome C (Cyt-C) and mitochondrial calcium in the testis of NIDDM rats. METHODS: Fifty male Wistar rats were divided randomly into three groups: model, ganoderma and normal control, the first two groups injected with 2% STZ through vena caudalis, and the last one with half-and-half sodium citrate/citrate buffer solution. Two weeks after normal diet, glucose tolerance tests were performed and the rats with abnormal glucose tolerance from the model and ganoderma groups received high-fat and high-carbohydrate food, the ganoderma group given Ganoderma lucidum spores (250mg/[ kg x d] ) in addition, both for 10 weeks. Glucose tolerance tests were repeated 1 day before the end of the experiment and the rats were castrated and relevant indexes measured. RESULTS: The NIDDM model was successfully constructed. In the model group, the levels of mitochondrial Cyt-C and mitochondrial calcium were significantly lower (P <0. 05) while that of the plasma Cyt-C was significantly higher than in the ganoderma and the control groups. CONCLUSION: Cyt-C and calcium ion are involved in the damage of the testis. Ganoderma lucidum spores can protect the testis of NIDDM rats.

[Study of the effect of super painkiller on the sexual function of male rats with type I diabetes].

OBJECTIVE: To study the effects of super painkiller on the sexual function of male Wistar rats with diabetes. METHODS: Thirty-two Wistar rats at 3-5 months of age were divided into two groups at random: 22 in the diabetes group, and 10 in the control group. After 72 hours, the former was further divided into 2 subgroups: non-treatment and super painkiller treatment. In 5 weeks, blood glucose was determined. After the apomorphines experiment, the rats were killed, blood taken from the vein, homogenate prepared from the isolated testis tissue and the level of NO and NOS in the serum and tissue homogenate surveyed, separately. RESULTS: Compared with the control group, serum NO and NOS of the diabetic rats dropped sharply. Compared with the non-treatment group, the serum NO and NOS of the super painkiller treatment group were very high and the difference was significant (P < 0.01). Apomorphine injection showed that the times of penis erection of the treated rats were more than those of the non-treatment group (P < 0.01) , but the difference was not significant compared with the control (P > 0.05). CONCLUSION: Super painkiller has sure effect of reducing blood glucose, with a certain curative value for sexual and reproductive malfunction of diabetic rats.