CPT1A mediates the succinylation of SP5 which activates transcription of PDPK1 to promote the viability and glycolysis of prostate cancer cells.

Succinylation modification involves in the progression of human cancers. The present study aimed to investigate the role of CPT1A, which is a succinyltransferase in the progression of prostate cancer (PCa). CCK-8 was used to detect the cell viability. Seahorse was performed to evaluate the cell glycolysis. Luciferase assay was used to detect the transcriptional regulation. ChIP was performed to assess the binding between transcriptional factors with the promoters. Co-IP was used to assess the binding between proteins. We found that CPT1A was highly expressed in PCa tissues and cell lines. Silencing of CPT1A inhibited the viability and glycolysis of PCa cells. Mechanistically, CPT1A promoted the succinylation of SP5, which strengthened the binding between SP5 and the promoter of PDPK1. SP5 activated PDPK1 transcription and PDPK1 activated the AKT/mTOR signal pathway. These findings might provide novel targets for the diagnosis or therapy of prostate cancer.

Radiofrequency ablation using the second-generation temperature-controlled diamond tip system in paroxysmal and persistent atrial fibrillation: results from FASTR-AF.

BACKGROUND: Catheter ablation (CA) technology development reflects the need to improve the effectiveness of atrial fibrillation (AF) treatment. Recently, the DiamondTemp Ablation (DTA) RF generator software was updated with a more responsive power ramp. METHODS: DIAMOND FASTR-AF was a prospective, single-arm, multicenter trial. This study sought to characterize the performance of the updated DTA system for the treatment of patients with drug-refractory paroxysmal and persistent AF (PAF and PsAF). The primary effectiveness endpoint was freedom from atrial arrhythmia recurrence following a 90-day blanking period through 12 months, and the primary safety endpoint was a composite of serious adverse events. RESULTS: In total, 60 subjects (34 PAF and 26 PsAF) underwent CA at three centers. Patients were 71.7% male, (age 63.9 ± 10.2 years, with an AF diagnosis duration 3.1 ± 3.9 years and left atrial size 4.4 ± 0.8 cm). Pulmonary vein isolation-only ablation strategy was performed in 34 (56.7%) subjects. The procedural characteristics show a procedure time 90.8 ± 31.6 min, total RF time 14.7 ± 7.7 min, ablation duration 10.7 ± 3.6 s, and fluid infusion 284.7 ± 111.5 ml. The serious adverse event rate was 8.3% (5/60), 3 pulmonary edema and 2 extended hospitalizations. Freedom from atrial arrhythmia recurrence was achieved in 67.6% of subjects by 12 months. CONCLUSIONS: The updated DTA system demonstrated long-term safety and effectiveness through 12 months of post-ablation follow-up for patients with atrial fibrillation. Additionally, procedures were demonstrated to be highly efficient with short procedure times and low levels of fluid infusion. TRIAL REGISTRATION: Sponsored by Medtronic, Inc.; FASTR-AF ClinicalTrials.gov; NCT03626649.

Treatment Efficacy of Continuous Renal Replacement on Symptoms, Inflammatory Mediators, and Coagulation Function in Patients with Sepsis-Associated Acute Kidney Injury.

INTRODUCTION: The aim of this study is to compare the treatment efficacy between continuous renal replacement therapy (CRRT) and conventional intermittent hemodialysis (IHD) in patients with sepsis (SIRS) combined with acute kidney injury (AKI) and its impact on inflammatory mediators and coagulation function. METHOD: 122 patients (25-60 years) with SIRS combined with AKI were enrolled in the sudy. The study group (SG) comprised 62 patients who received CRRT (8-10 h/day) + routine treatment, whereas the control group (CG) comprised 60 patients who received conventional IHD (4 h/day, 3 times per week) + routine treatment. inflammatory mediators and coagulation function measures were assessed and compared in each group. RESULTS: C-reactive protein, blood creatinine, blood urea nitrogen, blood lactic acid, oxygenation index, central venous oxygen saturation, SOFA (Sequential Organ Failure Assessment) score, interleukin 6, interleukin 8, hypersensitive C-reactive protein, tumor necrosis factor-α, prothrombin time, activated partial thromboplastin time, FIB, and platelet count were better in the SG than in the CG (p < 0.05). The 12- and 24-month survival rates were significantly higher in the SG than in the CG (p < 0.05). CONCLUSIONS: CRRT can effectively improve clinical symptoms, remove inflammatory factors, and maintain blood coagulation function in patients with SIRS combined with AKI. It is more efficient than IHD treatment and is worthy of clinical promotion.

Movement disorder caused by FRRS1L deficiency may be associated with morphological and functional disorders in Purkinje cells.

Ferric Chelate Reductase 1 Like (FRRS1L) protein has been identified as an auxiliary regulatory protein for the α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid receptor (AMPAR). FRRS1L is highly expressed in the cerebellum and other brain regions associated with the control of motor function. Loss of FRRS1L has been shown to lead to impaired synaptic transmission via AMPARs and to movement disorders. We found that deletion of the FRRS1L gene causes hyperactivity, reduced muscle strength, impaired coordination, and ataxia in mice. Deletion also impairs Purkinje cell dendritic spine formation and AMPAR expression in the cerebellum and damages the electrophysiological discharge rhythm of Purkinje cells. Cerebrospinal fluid examination and oleic acid (OA)-induced lipid accumulation monitoring in FRRS1L-knockdown SH-SY5Y cells indicated that FRRS1L deficiency could lead to aberrant metabolism of amino acids, glucose, and lipids. In summary, we found that the deletion of FRRS1L leads to impaired motor coordination and cerebellar ataxia in mice, which might be related to the reduced expression of AMPARs, metabolic deviations, and dysplastic functional defects in Purkinje cells.

Efficacy of continuous ambulatory peritoneal dialysis combined with hemodialysis versus single hemodialysis in patients with end-stage renal disease.

OBJECTIVE: This study aimed to compare the efficacy of single hemodialysis (HD) and continuous ambulatory peritoneal dialysis (CAPD) combined with HD in the treatment of end-stage renal disease. METHODS: Seventy patients with end-stage renal disease in our hospital from January 2019 to December 2020 were included and divided into 35 patients in the single group (SG) and 35 patients in the combination group (CG) according to a random number table. The SG received HD treatment and the CG received CAPD combined with HD treatment. RESULTS: Hemoglobin and serum albumin levels were higher, blood urea nitrogen (BUN) and serum creatinine (Scr) levels were lower, and interleukin-6 (IL-6), C-reactive protein (CRP), and tumor necrosis factor-α (TNF-α) levels were lower in the CG than in SG at the end of treatment (P < 0.05). Ca2+ levels were higher and P3+ levels were decreased at the end of treatment in both groups compared with those before treatment (P < 0.05), and Ca2+ and P3+ levels at the end of treatment in the CG were not different from those in the SG (P > 0.05). The complication rate in the CG was 5.71%, which was lower than 25.71% in the SG (P < 0.05). Quality of life scores were higher in the CG than in the SG at the end of treatment (P < 0.05). CONCLUSION: CAPD combined with HD can improve renal function and nutritional levels more significantly, control inflammatory responses more effectively, and reduce complications compared to single HD treatment in patients with end-stage renal disease.

Establishment of rat liver cancer cell lines with different metastatic potential.

The gloomy outcome of liver cancer is mainly due to the high rates of metastasis and recurrence, even after curative resection for early stage liver cancer. Our study was conducted to find the animal model suitable for the study of liver cancer metastasis. In our study, two liver cancer cells were obtained from N-nitrosodiethylamine (DEN) and N-nitrosomorpholine (NMOR) induced rats, and they were cultivated, screened and cloning cultivated. Bionomics of cells was analyzed. The results show that 2 cells had different metastatic potentiality. They were named Wrh-f2 and Wrh-s2, and they have the characteristics of Hepatocellular carcinoma cells. The bionomics of 2 cells showed: (1) The chromosome karyotype analysis showed that the mode of Wrh-f2 was 80-83 and Wrh-s2 was 55-57; (2) AFP positive cytoplasmic staining was observed in Wrh-f2 and Wrh-s2. Cytokeratin (CK) 7 and CK8 positive staining was present in Wrh-f2. CK8 positive staining was present in Wrh-s2; (3) The numbers of Wrh-f2 and Wrh-s2 that passed through the Transwells were 98 ± 12 and 55 ± 15;(4) Wrh-f2 had the significant higher colony formation (78%) than Wrh-s2(8%) (P < 0.01). (5) The animal models generated solid tumours when 2 cells were inoculated to nude mouse and rat. And Wrh-f2 developed stable pulmonary metastasis. The established cell lines with different metastatic potential showed obvious advantages over liver cancer in mimicking the biological properties of malignant liver cancer tumors. It provided a suitable model for the mechanism of liver cancer metastasis in vivo and in vitro.

Facile Synthesis of Self-Assembled NiFe Layered Double Hydroxide-Based Azobenzene Composite Films with Photoisomerization and Chemical Gas Sensor Performances.

Two kinds of layered double hydroxide (LDH) Langmuir composite films containing azobenzene (Azo) groups were successfully prepared by Langmuir-Blodgett (LB) technology. Then, an X-ray diffractometer (XRD), a transmission electron microscope (TEM), and an atomic force microscope (AFM) were used to investigate the structures of NiFe-LDH and the uniform morphologies of the composite LB films. The photoisomerization and acid-base gas sensor performances of the obtained thin film samples were tested by infrared visible (FTIR) spectroscropy and ultraviolet visible (UV-vis) spectroscropy. It is proved that the Azo dye molecules in the composite film are relatively stable to photoisomerization. In addition, the prepared composite films have high sensing sensitivity and good recyclability for acid-base response gases. The present research proposes a new clue for designing thin film materials for chemical gas response with good stability and sensitivity.

Facile Synthesis of Ag/Pd Nanoparticle-Loaded Poly(ethylene imine) Composite Hydrogels with Highly Efficient Catalytic Reduction of 4-Nitrophenol.

Poly(ethylene imine) (PEI) has abundant amino groups in a macromolecular chain and can be used as a graft source for metal nanocomposites, which shows excellent ability to form stable complexes with heavy metal ions. In this work, a simple and convenient method was used to make PEI into a stable hydrogel with 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide-N-hydroxysuccinimide and subsequently coprecipitate with silver nitrate solution or palladium chloride solution to form metal-loaded composite hydrogels. In addition, the characterizations of composite hydrogels were investigated by scanning electron microscopy, specific surface area tests (Brunauer-Emmett-Teller), X-ray photoelectron spectroscopy, and ultraviolet spectroscopy. The properties of composite hydrogels on the catalytic reduction of 4-nitrophenol were studied. The results showed that the composite hydrogels could be easily separated from the water environment, which indicated the large-scale potential application in organic catalytic degradation and wastewater treatment.

Self-assembled functional components-doped conductive polypyrrole composite hydrogels with enhanced electrochemical performances.

A conductive hydrogel is a composite conductive material formed by combining a conductive polymer with a nanogel structure of a hydrogel. Conductive hydrogels not only have potential applications in supercapacitors, sensors, and modulators, they can also be synthesized by many methods, such as copolymerization, crosslinking, and grafting. In this work, we successfully prepared three conductive composite hydrogels by in situ polymerization, namely polypyrrole sodium alginate conductive hydrogel, ferric chloride-doped polypyrrole sodium alginate hydrogel and doped polypyrrole sodium alginate hydrogel with sodium dodecylbenzene sulfonate. In addition, a series of characterizations were performed for the three conductive hydrogels described above. The results show that the polypyrrole sodium alginate hydrogel doped with ferric chloride forms a nanofiber network with a more stable structure and better electrochemical performance.

Simultaneous nitrification, denitrification and phosphorus removal in a sequencing batch reactor (SBR) under low temperature.

Simultaneous nitrogen and phosphorus removal in winter is one of the great challenges in wastewater treatment processes due to the poor bioactivity of microbial communities. In this study, excellent performance of simultaneous nitrification, denitrification and phosphorus removal (SNDPR) was achieved at low temperature of 10 °C and COD/N ratio of 6 in a lab-scale sequencing batch reactor. Total nitrogen (TN) and phosphorus (TP) removal efficiency reached 89.6% and 97.5%, respectively, accompanied with N2O emission of 7.46% TN due to the primary contribution (70%) of nitrifier denitrification. It was further confirmed that polyphosphate accumulating organisms (PAOs) were dominant in microbial communities revealed by fluorescence in situ hybridization and 16S rRNA amplicon sequencing. Moreover, denitrifying phosphorus removal by PAOs through nitrite pathway was found to be the main reason for the high efficiency of this SNDPR process. Denitrifying PAOs, especially the subgroup PAOII capable of utilizing nitrite to take up phosphorus, played a significant role in highly efficient TN and TP removal at low temperature. Furthermore, genus Propionivibrio was enriched (48.9%) in the bacterial community based on the 16S rRNA analysis, which was proposed to be a crucial member involved in the nitrogen and phosphorus removal simultaneously at low temperature in this system.

[Ressarch Advances of Prevantive and Therapeatic Effect of Mesenchymal Stromal Cells on Graft-Versus-Host Disease--Review].

OBJECTIVE: Graft-versus-host disease (GVHD) is a frequently encountered serious complication after allogeneic hematopoietic stem cell transplantation (allo-HSCT), it limits the success and widespread use of allo-HSCT. Mesenchymal stem cells (MSCs) are selected as ideal prophylactic and treatment means for GVHD during allo-HSCT due to their unique immunomodulatory and regenerative properties. Herein, the recent research progress about the prevantive and therapeutic effects of MSCs on GVHD and several issues related with the applications of MSC, including whether MSCs increasing risk of primary disease relapse and infection, impact of several clinical parameters on the clinical response to MSCs, and the prevantive and therapeutic effect of MSC-derived extracellular vesicles on GVHD are systematically reviewed.

Responsive surface bioaffinity binding to construct flexible and sensitive electrochemical aptasensors.

The development of simple, flexible, cost-effective and sensitive electrochemical biosensing strategies is highly desirable to advance their applications in disease diagnostics and clinical biomedicine. Herein, we fabricated a new enzyme-based electrochemical aptasensor with the use of adenosine triphosphate (ATP) and thrombin as model targets on the basis of a responsive surface bioaffinity binding strategy. It took full advantage of an immobilized complex duplex probe (hybrids of a hairpin-like aptamer probe with a digoxigenin (Dig)-labeled immobilization strand) to effectively inhibit the approach of the bulky horseradish peroxidase linked-anti-Dig antibody (anti-Dig-HRP) to the Dig on the electrode due to the steric effect. The target recognition dissociated the aptamer strand from the duplex probe and exposed the Dig for its effective binding with anti-Dig-HRP. The successive electrocatalysis offered a significantly amplified electrochemical signal correlated with the target recognition event. Sensitive detection toward ATP and thrombin was achieved with detection limits of 0.87 nM and 6.3 pM, respectively. The proposed strategy is simple and sensitive without any complex operations that hinder many amplified aptasensors. Also, the target recognition and signal reporting units are relatively isolated, making the biosensor fabrication more flexible. It thus provided a new and versatile pathway for sensitive biosensor fabrication.

Single- Versus Double-Unit Umbilical Cord Blood Transplantation for Hematologic Diseases: A Systematic Review.

Controversial results exist regarding the clinical benefits of single- vs double-unit umbilical cord blood transplantation (UCBT) in patients with hematologic diseases. A systematic review was conducted to evaluate this issue. The PubMed, Embase, and Cochrane Library databases were searched up to May 2018. A total of 25 studies including 6571 recipients were identified. Although double-unit UCB contained higher doses of total nucleated cells and CD34+ cells, it offered no advantages over single-unit UCB in terms of hematologic recovery, including the rate and speed of neutrophil and platelet engraftment. Double-unit UCBT was associated with higher incidences of grades II-IV acute and extensive chronic graft-vs-host disease, accompanied by a lower relapse incidence, which may be attributed to a graft-vs-graft effect induced by double-unit UCB. However, transplant-related mortality, disease-free survival, and overall survival were comparable between single- and double-unit UCBT. Although double-unit UCBT confers no clinical advantages over single-unit UCBT, certain patients, such as those at high risk of relapse, might benefit from double-unit UCBT, a possibility that needs to be clarified in future randomized trials.

The venom of spider Haplopelma hainanum suppresses proliferation and induces apoptosis in hepatic cancer cells by caspase activation in vitro.

ETHNOPHARMACOLOGICAL RELEVANCE: Spiders and spider venoms have been used in traditional Chinese medicine to treat various ailments for more than 1000 years. For instance, several large spiders have been utilized by the Li People, who mainly live in Hainan Island of China, in their own unique traditional Chinese medicine therapy. Recent studies have indicated that spider venoms may be an important source of bioactive compounds for anti-tumor treatments. However, the specific mechanisms underlying these activities are not yet completely understood. AIM OF THE STUDY: The present study investigated how the venom of the spider Haplopelma hainanum regulate proliferation and apoptosis in HepG2 cells via the underlying molecular mechanisms. MATERIALS AND METHODS: We treated HepG2 cells with various concentrations of the spider venom (0, 10, 50, 100 and 200 µg/mL) for 48 h, and then analyzed anti-proliferation activity, apoptosis-inducing effects, mitochondrial membrane potential (Δψm) and changes in the pro-apoptotic pathway. The anti-proliferation activity was detected by the MTT assay and Western blotting. Flow cytometry was used to analyze both apoptosis and mitochondrial membrane potential. The key pro-apoptotic molecules in the caspase-3 and -9 dependent mitochondrial pathway, including Bcl2 family, were assessed through realtime PCR, Western blotting and enzymatic test. RESULTS: Obvious morphological changes induced by the spider venom included decreased cell numbers, shorter cell length and reduced cell adhesion. MTT and Western blotting demonstrated that the spider venom potently suppressed cell proliferation in a dose- and time-dependent manner with IC50 of 126.00 µg/mL for 48 h. In addition, the spider venom caused a reduction in the mitochondrial membrane potential and cytochrome c release from mitochondria to cytoplasm under the participation of Bax. Finally, cytochrome c activated caspase-3 and caspase-9, and induced the apoptosis in the HepG2 cells. CONCLUSION: The results indicated that the venom of H. hainanum exhibited potent inhibition effects in HepG2 cells through suppressing proliferation, reducing the mitochondrial membrane potential, activating caspase-3 and caspase-9, and inducing the apoptosis through a mitochondrial-dependent pathway.

Efficacy and safety of mesenchymal stromal cells for the prophylaxis of chronic graft-versus-host disease after allogeneic hematopoietic stem cell transplantation: a meta-analysis of randomized controlled trials.

A meta-analysis of randomized controlled trials (RCTs) was conducted to evaluate the efficacy and safety of mesenchymal stromal cells (MSCs) for the prophylaxis of chronic graft-versus-host disease (cGVHD) in patients with hematological malignancies undergoing allogeneic hematopoietic stem cell transplantation (allo-HSCT). Six studies involving 365 patients were included. The pooled results showed that MSCs significantly reduced the incidence of cGVHD (risk ratio [RR] 0.63, 95% confidence interval [CI] 0.46 to 0.86, P = 0.004). Favorable prophylactic effects of MSCs on cGVHD were observed with umbilical cord-derived, high-dose, and late-infusion MSCs, while bone marrow-derived, low-dose, and coinfused MSCs did not confer beneficial prophylactic effects. In addition, MSC infusion did not increase the risk of primary disease relapse and infection (RR 1.02, 95% CI 0.70 to 1.50, P = 0.913; RR 0.89, 95% CI 0.44 to 1.81, P = 0.752; respectively). Moreover, there was an apparent trend toward increased overall survival (OS) in the MSC group compared with that in the control group (RR 1.13, 95% CI 0.98 to 1.29, P = 0.084). In conclusion, this meta-analysis demonstrated that MSC infusion is an effective and safe prophylactic strategy for cGVHD in patients with hematological malignancies undergoing allo-HSCT.

Genomic insights into metabolic potentials of two simultaneous aerobic denitrification and phosphorus removal bacteria, Achromobacter sp. GAD3 and Agrobacterium sp. LAD9.

Bacteria capable of simultaneous aerobic denitrification and phosphorus removal (SADPR) are promising for the establishment of novel one-stage wastewater treatment systems. Nevertheless, insights into the metabolic potential of SADPR-related bacteria are limited. Here, comprehensive metabolic models of two efficient SADPR bacteria, Achromobacter sp. GAD3 and Agrobacterium sp. LAD9, were obtained for the first time by high-throughput genome sequencing. With succinate as the preferred carbon source, both strains employed a complete TCA cycle as the major carbon metabolism for potentials of various organic acids and complex carbon oxidation. Complete and truncated aerobic denitrification routes were confirmed in GAD3 and LAD9, respectively, facilitated by all the major components of the electron transfer chain via oxidative phosphorylation. Comparative genome analysis revealed distinctive ecological niches involved in denitrification among different phylogenetic clades within Achromobacter and Agrobacterium. Excellent phosphorus removal capacities were contributed by inorganic phosphate uptake, polyphosphate synthesis and phosphonate metabolism. Additionally, the physiology of GAD3/LAD9 is different from that displayed by most available polyphosphate accumulating organisms, and reveals both strains to be more versatile, carrying out potentials for diverse organics degradation and outstanding SADPR capacity within a single organism. The functional exploration of SADPR bacteria broadens their significant prospects for application in concurrent aerobic carbon and nutrient removal.

[Role of Intestinal Microbiota in Hematopoietic Stem Cell Transplantation - Review].

Graft-versus-host disease (GVHD) and infection are the frequently encountered complications after hematopoietic stem cell transplantation (HSCT), which influence the outcome and limit the widespread application of HSCT. Intestinal microbiota plays an important role in maintaining intestinal immune balance. The diverse levels of intestinal microbiota associated with the incidences of GVHD, infection and the prognosis of HSCT, thus remodeling intestinal microbiota can alleviate GVHD and infection after HSCT. Herein, the recent research progress about the role and the involved mechanisms of intestinal microbiota in HSCT, and the novel manipulation strategies of intestinal microbiota are systematically reviewed.

Regulatory crosstalk between KLF5, miR-29a and Fbw7/CDC4 cooperatively promotes atherosclerotic development.

Atherogenesis is a chronic inflammatory process that involves complex interactions between endothelial dysfunction, lipid deposition and vascular smooth-muscle cell (VSMC) proliferation. However, the molecular mechanism is still unclear. We found that a pro-atherosclerotic factor (oxLDL) induced the expression of Krüppel-like factor 5 (KLF5), which in turn increased miR-29a expression levels. The increased miR-29a was retained within HASMCs and down-regulated Fbw7/CDC4 expression by targeting the 3´UTR of Fbw7/CDC4, subsequently increasing KLF5 stability by reducing the Fbw7/CDC4-dependent ubiquitination of KLF5, forming a positive feedback loop to enhance VSMC proliferation and promote atherogenesis. These results indicate a potentially important role for the oxLDL-activated feedback mechanism in VSMC proliferation and atherogenesis. Suppression of miR-29a may be an effective way to attenuate atherosclerosis. In conclusion, our data are the first to reveal that the regulatory crosstalk between KLF5, miR-29a, and Fbw7/CDC4 cooperatively promotes atherosclerotic development.

The expression and construction of engineering Escherichia coli producing humanized AluY RNAs.

BACKGROUND: Exogenous RNAs can specifically up-regulate or down-regulate gene expression after they enter into cells. Alu RNAs are the main constituent of human transcriptome and participate in gene expression regulation. AluY elements belong to a subfamily of Alus and are the youngest Alus. In this paper, we established the technology method of preparing genetically engineered humanized AluY RNAs (AluY RNAs) from Escherichia coli (E. coli) strains. This technology method also can be used to prepare other genetically engineered humanized RNAs that can be used for cytology experiments. RESULTS: Different copies of human AluY elements were inserted into pET-28α plasmid (pET) to construct pET-AluY plasmids that were transformed into BMBL21-DE3 (DE3) E. coli. Isopropylthio-ß-D-galactoside (IPTG) induction inhibited transformed bacterial growth after DE3 E. coli were transformed by pET-AluY × 8 plasmid (8 copies of AluYs were inserted into pET); northern blotting was used to detect the amount of AluY RNAs after 2, 4, 6, 8, 10, 12, 14 and 16 h inducing with IPTG. The results showed that the amount of AluY RNAs was the highest at 4 h; 1, 2, 4, 8 or 14 copies of AluY elements were inserted into the pET to construct pET-AluY plasmids that were transformed into DE3 bacteria, the northern blotting results showed that AluY RNAs production amount increased with the increase of AluY copy number; pET-AluY × 8 DE3 bacteria did not produce AluY RNAs without IPTG induction, AluY RNA production kept similar when inducing by 0.1-0.4 mg/ml IPTG induction, however, AluY RNA production slightly decreased if deviating from the above concentration range; pET-AluY × 8 DE3 bacteria were cultured at 34, 37 or 40 °C and the results showed that AluY RNA production was the highest under 37 °C cultivation; pET-AluY × 8 plasmid was transformed into three kinds of BL21 bacteria, including DE3, BMBL21-DE3-pLysS (pLysS) and Trans BL 21 (TransBL), the results showed that AluY RNA production was the highest when using DE3 bacteria. CONCLUSIONS: The optimal conditions of producing AluY RNAs were: a kind of host bacteria of DE3, an engineering bacteria concentration of OD600 1.0, an IPTG concentration of 0.2 mg/ml, a culturing temperature of 37 °C and a culturing time of 4 h. Pure AluY RNAs occupied 15.8% of extractive total RNAs and the mean yield of pure AluY RNAs in 100 ml bacteria solution was 0.46 mg.

An Integrative Analysis Reveals a Central Role of P53 Activation via MDM2 in Zika Virus Infection Induced Cell Death.

Zika virus (ZIKV) infection is an emerging global threat that is suspected to be associated with fetal microcephaly. However, the molecular mechanisms underlying ZIKV disease pathogenesis in humans remain elusive. Here, we investigated the human protein interaction network associated with ZIKV infection using a systemic virology approach, and reconstructed the transcriptional regulatory network to analyze the mechanisms underlying ZIKV-elicited microcephaly pathogenesis. The bioinformatics findings in this study show that P53 is the hub of the genetic regulatory network for ZIKV-related and microcephaly-associated proteins. Importantly, these results imply that the ZIKV capsid protein interacts with mouse double-minute-2 homolog (MDM2), which is involved in the P53-mediated apoptosis pathway, activating the death of infected neural cells. We also found that synthetic mimics of the ZIKV capsid protein induced cell death in vitro and in vivo. This study provides important insight into the relationship between ZIKV infection and brain diseases.