Study on the Shear Strength and Erosion Resistance of Sand Solidified by Enzyme-Induced Calcium Carbonate Precipitation (EICP).

Sand solidification of earth-rock dams is the key to flood discharge capacity and collapse prevention of earth-rock dams. It is urgent to find an economical, environmentally friendly, and durable sand solidification technology. However, the traditional grouting reinforcement method has some problems, such as high costs, complex operations, and environmental pollution. Enzyme-induced calcium carbonate precipitation (EICP) is an anti-seepage reinforcement technology emerging in recent years with the characteristics of economy, environmental protection, and durability. The erosion resistance and shear strength of earth-rock dams solidified by EICP need further verification. In this paper, EICP-solidified standard sand is taken as the research object, and EICP-cemented standard sand is carried out by a consolidated undrained triaxial test. A two-stage pouring method is adopted to pour samples, and the effects of dry density, cementation times, standing time, and confining pressure on the shear strength of cemented standard sand are emphatically analyzed. The relationship between cohesion, internal friction angle, and CaCO3 formation was analyzed. After the optimal curing conditions are obtained through the triaxial shear strength test, the erosion resistance model test is carried out. The effects of erosion angle, erosion flow rate, and erosion time on the erosion resistance of EICP-solidified sand were analyzed through an erosion model test. The results of triaxial tests show that the standard sand solidified by EICP exhibits strain softening, and the peak strength increases with the increase in initial dry density, cementation times, standing time, and confining pressure. When the content of CaCO3 increases from 2.84 g to 12.61 g, the cohesive force and internal friction angle change to 23.13 times and 1.18 times, and the determination coefficients reach 0.93 and 0.94, respectively. Erosion model test results indicate that the EICP-solidified sand dam has good erosion resistance. As the increase in erosion angle, erosion flow rate, and erosion time, the breach of solidified samples gradually becomes larger. Due to the deep solidification of sand by EICP, the development of breaches is relatively slow. Under different erosion conditions, the solidified samples did not collapse and the dam broke. The research results have important reference value and scientific significance for the practice of sand consolidation engineering in earth-rock dams.

Identification of novel immune-related signatures for keloid diagnosis and treatment: insights from integrated bulk RNA-seq and scRNA-seq analysis.

BACKGROUND: Keloid is a disease characterized by proliferation of fibrous tissue after the healing of skin tissue, which seriously affects the daily life of patients. However, the clinical treatment of keloids still has limitations, that is, it is not effective in controlling keloids, resulting in a high recurrence rate. Thus, it is urgent to identify new signatures to improve the diagnosis and treatment of keloids. METHOD: Bulk RNA seq and scRNA seq data were downloaded from the GEO database. First, we used WGCNA and MEGENA to co-identify keloid/immune-related DEGs. Subsequently, we used three machine learning algorithms (Randomforest, SVM-RFE, and LASSO) to identify hub immune-related genes of keloid (KHIGs) and investigated the heterogeneous expression of KHIGs during fibroblast subpopulation differentiation using scRNA-seq. Finally, we used HE and Masson staining, quantitative reverse transcription-PCR, western blotting, immunohistochemical, and Immunofluorescent assay to investigate the dysregulated expression and the mechanism of retinoic acid in keloids. RESULTS: In the present study, we identified PTGFR, RBP5, and LIF as KHIGs and validated their diagnostic performance. Subsequently, we constructed a novel artificial neural network molecular diagnostic model based on the transcriptome pattern of KHIGs, which is expected to break through the current dilemma faced by molecular diagnosis of keloids in the clinic. Meanwhile, the constructed IG score can also effectively predict keloid risk, which provides a new strategy for keloid prevention. Additionally, we observed that KHIGs were also heterogeneously expressed in the constructed differentiation trajectories of fibroblast subtypes, which may affect the differentiation of fibroblast subtypes and thus lead to dysregulation of the immune microenvironment in keloids. Finally, we found that retinoic acid may treat or alleviate keloids by inhibiting RBP5 to differentiate pro-inflammatory fibroblasts (PIF) to mesenchymal fibroblasts (MF), which further reduces collagen secretion. CONCLUSION: In summary, the present study provides novel immune signatures (PTGFR, RBP5, and LIF) for keloid diagnosis and treatment, and identifies retinoic acid as potential anti-keloid drugs. More importantly, we provide a new perspective for understanding the interactions between different fibroblast subtypes in keloids and the remodeling of their immune microenvironment.

Experimental Study on the Crack Concrete Repaired via Enzyme-Induced Calcium Carbonate Precipitation (EICP).

A low-carbon and environmentally friendly EICP method for repairing concrete cracks is presented to prolong the service life of concrete. In this study, we took concrete as the research object and quartz sand as the filling medium and employed the EICP injection method to repair concrete cracks. The internal repair effect of EICP on concrete cracks was evaluated with a combination of ultrasonic and compressive strength tests. The concrete repair mechanism of EICP was identified with a combination of EDS, XRD, and SEM tests. The results indicate that with an increase in the fracture depth, the ultrasonic sound time of the crack specimen increased gradually, and the ultrasonic wave transit time value of the crack specimen decreased significantly after EICP repair. After repair, the compressive strength rose. The highest compressive-strength recovery rate of a 0.3 mm wide specimen is 98.41%. The calcium carbonate crystal formed using EICP is vaterite. The probability density function model of the Laplace distribution was constructed, which showed good applicability and consistency in the ultrasonic sound time and compressive strength measured via experiments. The formed calcium carbonate crystals can be tightly and evenly attached to the cracks with the EICP injection repair method, resulting in a better repair effect.

Study on the Mechanical Properties of Crack Mortar Repaired by Enzyme-Induced Calcium Carbonate Precipitation (EICP).

As an emerging repair method, the enzyme-induced calcium carbonate precipitation (EICP) technique has the advantages of being highly economical, eco-friendly, and durable. The optimal repair conditions were obtained by taking cement mortar as the research object, adding two types of filling medium, using three EICP-based repair methods to repair the cement mortar with different crack widths, and combining ultrasonic testing and strength testing to evaluate the mechanical properties and repair effects of the repair mortar. The microscopic structure of the mortar was established using mesoscopic and microscopic tests (XRD, SEM, and EDS), thereby revealing the mechanism of repair based on EICP. The test results show that, when quartz sand is used as the repair medium, more calcium carbonate adheres to the cross-section of test samples, and it has a better repair effect. Moreover, the repair effect of the injection method is significantly higher than those of the perfusion and immersion methods, and the ultrasonic wave transit time decreases by 1.22% on average. Based on the combination of quartz sand and EICP repair methods, the calcium carbonate precipitated among the sand granules contributes to a binding effect that strengthens the cohesive force among the sand granules.

Chromosome-level genome assembly of the bethylid ectoparasitoid wasp Sclerodermus sp. 'alternatusi'.

The Bethylidae are the most diverse of Hymenoptera chrysidoid families. As external parasitoids, the bethylids have been widely adopted as biocontrol agents to control insect pests worldwide. Thus far, the genomic information of the family Bethylidae has not been reported yet. In this study, we crystallized into a high-quality chromosome-level genome of ant-like bethylid wasps Sclerodermus sp. 'alternatusi' (Hymenoptera: Bethylidae) using PacBio sequencing as well as Hi-C technology. The assembled S. alternatusi genome was 162.30 Mb in size with a contig N50 size of 3.83 Mb and scaffold N50 size of 11.10 Mb. Totally, 92.85% assembled sequences anchored to 15 pseudo-chromosomes. A total of 10,204 protein-coding genes were annotated, and 23.01 Mb repetitive sequences occupying 14.17% of genome were pinpointed. The BUSCO results showed that 97.9% of the complete core Insecta genes were identified in the genome, while 97.1% in the gene sets. The high-quality genome of S. alternatusi will not only provide valuable genomic information, but also show insights into parasitoid wasp evolution and bio-control application in future studies.

A computer-aid speech rehabilitation system with mirrored video generating.

BACKGROUND: Aphasia is a communication disorder that affects the ability to process and produce language, which severely impacting their lives. Computer-aid exercise rehabilitation has shown to be highly effective for these patients. OBJECTIVE: In our study, we proposed a speech rehabilitation system with mirrored therapy. The study goal is to construct a effective rehabilitation software for aphasia patients. METHODS: This system collects patients' facial photos for mirrored video generation and speech synthesis. The visual feedback provided by the mirror creates an engaging and motivating experience for patients. And the evaluation platform employs machine learning technologies for assessing speech similarity. RESULTS: The sophisticated task-oriented rehabilitation training with mirror therapy is also presented for experiments performing. The performance of three tasks reaches the average scores of 83.9% for vowel exercises, 74.3% for word exercies and 77.8% for sentence training in real time. CONCLUSIONS: The user-friendly application system allows patients to carry out daily training tasks instructed by the therapists or the prompt information of menu. Our work demonstrated a promising intelligent mirror software system for reading-based aphasia rehabilitation.

PTTH-Torso Signaling System Controls Developmental Timing, Body Size, and Reproduction through Regulating Ecdysone Homeostasis in the Brown Planthopper, Nilaparvata lugens.

In holometabolous insects, such as Drosophila and Bombyx, prothoracicotropic hormone (PTTH) is well established to be critical in controlling developmental transitions and metamorphosis by stimulating the biosynthesis of ecdysone in the prothoracic glands (PGs). However, the physiological role of PTTH and the receptor Torso in hemimetabolous insects remains largely unexplored. In this study, homozygous PTTH- and Torso-null mutants of the brown planthopper (BPH), Nilaparvata lugens, were successfully generated by employing clustered regularly interspaced short palindromic repeats/CRISPR-associated 9 (CRISPR-Cas9). Further characterization showed that both NlPTTH-/- and NlTorso-/- mutants exhibited prolonged nymphal duration and increased final adult size. Enzyme-linked immunosorbent assay (ELISA) revealed that NlPTTH-/- and NlTorso-/- mutants exhibited a significant reduction in 20-hydroxyecdysone (20E) in fifth-instar nymphs at 48 h post-ecdysis compared to Wt controls. Furthermore, our results indicated that both NlPTTH-/- and NlTorso-/- mutants had shortened lifespan, reduced female fecundity, and reduced egg hatching rates in adults. These findings suggest a conserved role for the PTTH-Torso signaling system in the regulation of developmental transitions by stimulating ecdysone biosynthesis in hemimetabolous insects.

miR-769-3p inhibits cellular proliferation of KSHV-infected SH-SY5Y cells through targeting mTOR.

The infection by Kaposi's sarcoma-associated herpesvirus (KSHV) is one of the most common causes of death in AIDS patients. Our studies have found that KSHV can infect SH-SY5Y cells (named SK-RG) in vivo and mTOR was up-regulated, which results in remarkable enhancement of cell proliferation, migration. But the regulatory role of mTOR in KSHV infected neurons has not yet been fully elucidated. Here, we find that miR-769-3p is decreased in SK-RG cells, which can exert anti-KSHV effect through negatively regulating the expression of mTOR. The knockdown of mTOR or overexpress of miR-769-3p decreased the proliferation, migration ability and cell cycle related protein of SK-RG cells, and the expression of KSHV related genes. In contrast, activating mTOR function by 3BDO treatment weakened the cellular behaviors of miR-769-3p overexpressing cells. Meanwhile, overexpressed miR-769-3p and rapamycin showed a shared inhibition trend in the effects on cell proliferation and motility. Our data indicated that miR-769-3p can inhibit cell proliferation and migration by down regulating mTOR in KSHV infected SH-SY5Y cells, and can be a candidate molecule for anti-KSHV therapy.

Dose-response associations of triglyceride to high-density lipoprotein cholesterol ratio and triglyceride-glucose index with arterial stiffness risk.

BACKGROUND: The triglyceride to high-density lipoprotein cholesterol (TG/HDL-C) ratio and triglyceride-glucose (TyG) index are novel indexes for insulin resistance (IR). We aimed to evaluate associations of TG/HDL-C and TyG with arterial stiffness risk. METHODS: We enrolled 1979 participants from the Rural Chinese Cohort Study, examining arterial stiffness by brachial-ankle pulse wave velocity (baPWV). Logistic and linear regression models were employed to calculate effect estimates. For meta-analysis, we searched relevant articles from PubMed, Embase and Web of Science up to August 26, 2023. The fixed-effects or random-effects models were used to calculate the pooled estimates. We evaluated dose-response associations using restricted cubic splines. RESULTS: For cross-sectional studies, the adjusted ORs (95%CIs) for arterial stiffness were 1.12 (1.01-1.23) and 1.78 (1.38-2.30) for per 1 unit increment in TG/HDL-C and TyG. In the meta-analysis, the pooled ORs (95% CIs) were 1.26 (1.14-1.39) and 1.57 (1.36-1.82) for per 1 unit increment of TG/HDL-C and TyG. Additionally, both TG/HDL-C and TyG were positively related to PWV, with ß of 0.09 (95% CI 0.04-0.14) and 0.57 (95% CI 0.35-0.78) m/s. We also found linear associations of TG/HDL-C and TyG with arterial stiffness risk. CONCLUSIONS: High TG/HDL-C and TyG were related to increased arterial stiffness risk, indicating TG/HDL-C and TyG may be convincing predictors of arterial stiffness.

Experimental Study on the Wind Erosion Resistance of Aeolian Sand Solidified by Microbially Induced Calcite Precipitation (MICP).

Microbially induced calcite precipitation (MICP) is an emerging solidification method characterized by high economic efficiency, environmental friendliness, and durability. This study validated the reliability of the MICP sand solidification method by conducting a small-scale wind tunnel model test using aeolian sand solidified by MICP and analyzing the effects of wind velocity (7 m/s, 10 m/s, and 13 m/s), deflation angle (0°, 15°, 30°, and 45°), wind erosion cycle (1, 3, and 5), and other related factors on the mass loss rate of solidified aeolian sand. The microstructure of aeolian sand was constructed by performing mesoscopic and microscopic testing based on X-ray diffraction analysis (XRD), Fourier-transform infrared spectroscopy (FTIR), and scanning electron microscopy (SEM). According to the test results, the mass loss rate of solidified aeolian sand gradually increases with the increase in wind velocity, deflation angle, and wind erosion cycle. When the wind velocity was 13 m/s, the mass loss rate of the aeolian sand was only 63.6%, indicating that aeolian sand has excellent wind erosion resistance. CaCO3 crystals generated by MICP were mostly distributed on sand particle surfaces, in sand particle pores, and between sand particles to realize the covering, filling, and cementing effects.

CD-Net: Cascaded 3D Dilated convolutional neural network for pneumonia lesion segmentation.

COVID-19 is a global pandemic that has caused significant global, social, and economic disruption. To effectively assist in screening and monitoring diagnosed cases, it is crucial to accurately segment lesions from Computer Tomography (CT) scans. Due to the lack of labeled data and the presence of redundant parameters in 3D CT, there are still significant challenges in diagnosing COVID-19 in related fields. To address the problem, we have developed a new model called the Cascaded 3D Dilated convolutional neural network (CD-Net) for directly processing CT volume data. To reduce memory consumption when cutting volume data into small patches, we initially design a cascade architecture in CD-Net to preserve global information. Then, we construct a Multi-scale Parallel Dilated Convolution (MPDC) block to aggregate features of different sizes and simultaneously reduce the parameters. Moreover, to alleviate the shortage of labeled data, we employ classical transfer learning, which requires only a small amount of data while achieving better performance. Experimental results conducted on the different public-available datasets verify that the proposed CD-Net has reduced the negative-positive ratio and outperformed other existing segmentation methods while requiring less data.

Indolent growth of low­grade myofibroblastic sarcoma of the knee with the resemblance of a benign lesion: A case report.

Low-grade myofibroblastic sarcoma (LGMS) is a rare soft-tissue malignant neoplasm originating from the stromal cells that is predominantly comprised of atypical myofibroblasts. The present study reports the case of a patient with low-grade myofibroblastic sarcoma in the soft tissue of the knee. The patient exhibited a soft, painless mass on the medial side of the left knee. During surgery, a yellow-white mass above the posterior margin of the sartorius muscle was identified. This mass, which was irregular and adherent to surrounding tissues, had a blurry boundary, indicative of invasive growth. The morphology of the mass significantly differed from conventional lipomas and was lacking typical features. Immunohistochemical staining confirmed the diagnosis of a malignant LGMS. The patient experienced no recurrence over 1 year of follow-up and reported complete recovery of knee range of motion. The present study suggests that an incisional biopsy may be performed if LGMS is suspected based on patient symptoms and imaging results. In addition, long-term follow-up is crucial for the timely detection and management of any recurrence, playing a key role in the patient's ongoing care and treatment.

Miscarriage and stillbirth in relation to risk of cardiovascular diseases: A systematic review and meta-analysis.

INTRODUCTION: The relationship between pregnancy loss and the risk of cardiovascular diseases (CVDs) remains a matter of debate. Our intention in conducting this meta-analysis was to analyze the relationship between miscarriage and stillbirth and risk of CVDs. METHODS: PubMed, Embase, and Web of Science were systematically searched up to May 30, 2023 for all relevant studies. The random-effects model was applied to estimate the pooled relative risks (RRs) and 95% confidence intervals (95% CIs). We evaluated RR estimates for the risk of CVDs with each additional miscarriage and stillbirth through generalized least squares regression. RESULTS: Twenty-three articles were incorporated into the meta-analysis. For women with a history of miscarriage, the pooled RRs for the risk of total CVDs, coronary heart disease (CHD), stroke, and total CVD deaths were 1.16 (95 % CI 1.10-1.22), 1.26 (1.12-1.41), 1.13 (1.03-1.24), and 1.20 (1.01-1.42), respectively. For women with a history of stillbirth, the pooled RRs for the risk of total CVDs, CHD, stroke, and total CVD deaths were 1.60 (1.34-1.89), 1.30 (1.12-1.50), 1.37 (1.06-1.78), and 1.95 (1.05-3.63), respectively. With each additional miscarriage, the risk increased for total CVDs (1.08, 1.04-1.13), CHD (1.08, 1.04-1.13), and stroke (1.05, 1.00-1.10). With each additional stillbirth, the risk increased for total CVDs (1.11, 1.03-1.21) and CHD (1.13, 1.07-1.19). CONCLUSION: This meta-analysis indicates that both miscarriages and stillbirths are related to a higher risk of total CVDs, CHD, stroke, and total CVD deaths. The risk of total CVDs and CHD increased with the number of miscarriages or stillbirths.

Obesity and risk of depressive disorder in children and adolescents: A meta-analysis of observational studies.

PURPOSE: This meta-analysis evaluated the relationship between overweight/obesity and depressive disorders in children and adolescents. METHODS: We examined the databases of PubMed, Embase and Web of Science for pertinent observational studies released up until 20 February 2022. The pooled relative risks (RRs) and 95% confidence intervals (CIs) of obesity and overweight with depressive disorder were calculated by means of random-effects models. The Newcastle-Ottawa Quality Assessment Scale and Agency for Healthcare Research and Quality scale were adopted to evaluate the study quality. RESULTS: Finally, for this meta-analysis, we evaluated 22 observational publications covering 175 135 participants (5 cohort study articles, 1 case-control study article and 16 cross-sectional study articles). A significant positive association was found between obesity and the risk of depression (RR 1.32, 95% CI 1.09-1.60, I2 = 79.90%, Pheterogeneity < 0.001) and in the association between obesity and depressive symptoms (RR 1.16, 95% CI: 1.00-1.35, I2 = 25.0%, Pheterogeneity = 0.247). On sensitivity analysis, the pooled RRs remained robust. Subgroup analysis indicated that obese children and teenagers in western countries were more prone to depression. CONCLUSION: Evidence from this meta-analysis, based on observational studies, supported the idea that obese children and adolescents are more likely to experience depression and depressive symptoms.

An alfalfa MYB-like transcriptional factor MsMYBH positively regulates alfalfa seedling drought resistance and undergoes MsWAV3-mediated degradation.

Drought is a major threat to alfalfa (Medicago sativa L.) production. The discovery of important alfalfa genes regulating drought response will facilitate breeding for drought-resistant alfalfa cultivars. Here, we report a genome-wide association study of drought resistance in alfalfa. We identified and functionally characterized an MYB-like transcription factor gene (MsMYBH), which increases the drought resistance in alfalfa. Compared with the wild-types, the biomass and forage quality were enhanced in MsMYBH overexpressed plants. Combined RNA-seq, proteomics and chromatin immunoprecipitation analysis showed that MsMYBH can directly bind to the promoters of MsMCP1, MsMCP2, MsPRX1A and MsCARCAB to improve their expression. The outcomes of such interactions include better water balance, high photosynthetic efficiency and scavenge excess H2O2 in response to drought. Furthermore, an E3 ubiquitin ligase (MsWAV3) was found to induce MsMYBH degradation under long-term drought, via the 26S proteasome pathway. Furthermore, variable-number tandem repeats in MsMYBH promoter were characterized among a collection of germplasms, and the variation is associated with promoter activity. Collectively, our findings shed light on the functions of MsMYBH and provide a pivotal gene that could be leveraged for breeding drought-resistant alfalfa. This discovery also offers new insights into the mechanisms of drought resistance in alfalfa.

Methylseleninic acid inhibits human glioma growth in vitro and in vivo by triggering ROS-dependent oxidative damage and apoptosis.

Selenium-containing agents showed novel anticancer activity by triggering pro-oxidative mechanism. Studies confirmed that methylseleninic acid (MeSe) displayed broad-spectrum anti-tumor activity against kinds of human cancers. However, the anticancer effects and mechanism of MeSe against human glioma growth have not been explored yet. Herein, the present study showed that MeSeA dose-dependently inhibited U251 and U87 human glioma cells growth in vitro. Flow cytometry analysis indicated that MeSe induced significant U251 cells apoptosis with a dose-dependent manner, followed by the activation of caspase-7, caspase-9 and caspase-3. Immunofluorescence staining revealed that MeSe time-dependently caused reactive oxide species (ROS) accumulation and subsequently resulted in oxidative damage, as convinced by the increased phosphorylation level of Ser428-ATR, Ser1981-ATM, Ser15-p53 and Ser139-histone. ROS inhibition by glutathione (GSH) effectively attenuated MeSe-induced ROS generation, oxidative damage, caspase-3 activation and cytotoxicity, indicating that ROS was an upstream factor involved in MeSe-mediated anticancer mechanism in glioma. Importantly, MeSe administration in nude mice significantly inhibited glioma growth in vivo by inducing apoptosis through triggering oxidative damage. Taken together, our findings validated the possibility that MeSe as a selenium-containing can act as potential tumor chemotherapy agent for therapy of human glioma.

A Ru/RuO2 heterostructure boosting electrochemistry-assisted selective benzoic acid hydrogenation.

Electrocatalytic hydrogenation of benzoic acid (BA) to cyclohexanecarboxylic acid (CCA) at ambient temperature and pressure has been recognized as a promising alternative to thermal hydrogenation since water is required as the hydrogen source. So far, only a few Pt-based electrocatalysts have been developed in acidic electrolyte. To overcome the limitations of reactant solubility and catalyst corrosion, herein, carbon fiber-supported Ru electrocatalysts with abundant Ru/RuO2 heterojunctions were fabricated via cyclic electrodeposition between -0.8 and 1.1 V vs. Ag/AgCl. In an alkaline environment, a Ru/RuO2 catalyst achieves an excellent ECH reactivity in terms of high BA conversion (100%) and selectivity towards CCA (100%) within 180 min at a current density of 200/3 mA cm-2, showing exceptional reusability and long-term stability. 1-Cyclohexenecarboxylic acid (CEA) was identified as the reaction intermediate, whose the selectivity is governed by the applied potential. Kinetic studies demonstrate that ECH of BA over Ru/RuO2 follows a Langmuir-Hinshelwood (L-H) mechanism. In situ Raman spectroscopy and theoretical calculations reveal that the Ru/RuO2 interface enhances the adsorption strength of CEA, thereby facilitating the production of fully hydrogenated CCA. This work provides a deep understanding of the ECH pathway of BA in alkaline media, and gives a new methodology to fabricate heterostructure electrocatalysts.

An Essential Role of c-Fos in Notch1-mediated Promotion of Proliferation of KSHV-Infected SH-SY5Y Cells.

BACKGROUND: This study aimed to investigate the influence of Notch1 on c-Fos and the effect of c-Fos on the proliferation of Kaposi's sarcoma-associated herpesvirus (KSHV)-infected neuronal cells. METHODS: Real-time PCR and western blotting were used to determine c-Fos expression levels in KSHV-infected (SK-RG) and uninfected SH-SY5Y cells. C-Fos levels were measured again in SK-RG cells with or without Notch1 knockdown. Next, we measured c-Fos and p-c-Fos concentrations after treatment with the Notch1 γ-secretase inhibitor LY-411575 and the Notch1 activator Jagged-1. MTT and Ki-67 staining were used to evaluate the proliferation ability of cells after c-Fos levels downregulation. CyclinD1, CDK6, and CDK4 expression levels and cell cycle were analyzed by western blotting and flow cytometry, respectively. After the c-Fos intervention, the KSHV copy number and gene expression of RTA, LANA and K8.1 were analyzed by real-time TaqMan PCR. RESULTS: C-Fos was up-regulated in KSHV-infected SK-RG cells. However, the siRNA-mediated knockdown of Notch1 resulted in a significant decrease in the levels of c-Fos and p-c-Fos (P <0.01, P <0.001). Additionally, a decrease in Cyclin D1, CDK6, and CDK4 was also detected. The Notch1 inhibitor LY-411575 showed the potential to down-regulate the levels of c-Fos and p-c-Fos, which was consistent with Notch1 knockdown group (P <0.01), whereas the expression and phosphorylation of c-Fos were remarkably up-regulated by treatment of Notch1 activator Jagged-1 (P <0.05). In addition, our data obtained by MTT and Ki-67 staining revealed that the c-Fos down-regulation led to a significant reduction in cell viability and proliferation of the SK-RG cells (P <0.001). Moreover, FACS analysis showed that the cell cycle was arrested in the G0/G1 stage, and the expressions of Cyclin D1, CDK6, and CDK4 were down-regulated in the c-Fos-knockdown SK-RG cells (P <0.05). Reduction in total KSHV copy number and expressions of viral genes (RTA, LANA and K8.1) were also detected in c-Fos down-regulated SK-RG cells (P <0.05). CONCLUSION: Our findings strongly indicate that c-Fos plays a crucial role in the promotion of cell proliferation through Notch1 signaling in KSHV-infected cells. Furthermore, our results suggest that the inhibition of expression of key viral pathogenic proteins is likely involved in this process.

Methylation in the promoter regions of WT1, NKX6-1 and DBC1 genes in cervical cancer tissues of Uygur women in Xinjiang

Abstract This study aimed to explore: 1) DNA methylation in the promoter regions of Wilms tumor gene 1 (WT1), NK6 transcription factor related locus 1 gene (NKX6-1) and Deleted in bladder cancer 1 (DBC1) gene in cervical cancer tissues of Uygur women in Xinjiang, and 2) the correlation of gene methylation with the infection of HPV16/18 viruses. We detected HPV16/18 infection in 43 normal cervical tissues, 30 cervical intraepithelial neoplasia lesions (CIN) and 48 cervical cancer tissues with polymerase chain reaction (PCR) method. Methylation in the promoter regions of the WT1, NKX6-1 and DBC1 genes in the above-mentioned tissues was measured by methylation-specific PCR (MSP) and cloning sequencing. The expression level of these three genes was measured by real-time PCR (qPCR) in 10 methylation-positive cervical cancer tissues and 10 methylation-negative normal cervical tissues. We found that the infection of HPV16 in normal cervical tissues, CIN and cervical cancer tissues was 14.0, 36.7 and 66.7%, respectively. The infection of HPV18 was 0, 6.7 and 10.4%, respectively. The methylation rates of WT1, NKX6-1 and DBC1 genes were 7.0, 11.6 and 23.3% in normal cervical tissues, 36.7, 46.7 and 30.0% in CIN tissues, and 89.6, 77.1 and 85.4% in cervical cancer tissues. Furthermore, WT1, NKX6-1 and DBC1 genes were hypermethylated in the high-grade squamous intraepithelial lesion (CIN2, CIN3) and in the cervical cancer tissues with infection of HPV16/18 (both P< 0.05). The expression of WT1, NKX6-1 and DBC1 was significantly lower in the methylation-positive cervical cancer tissues than in methylation-negative normal cervical tissues. Our findings indicated that methylation in the promoter regions of WT1, NKX6-1 and DBC1 is correlated with cervical cancer tumorigenesis in Uygur women. The infection of HPV16/18 might be correlated with methylation in these genes. Gene inactivation caused by methylation might be related to the incidence and development of cervical cancer.