Highly Porous 3D Printed Tantalum Scaffolds Have Better Biomechanical and Microstructural Properties than Titanium Scaffolds.

OBJECTIVE: To test the biomechanical properties of 3D printed tantalum and titanium porous scaffolds. METHODS: Four types of tantalum and titanium scaffolds with four alternative pore diameters, #1 (1000-700 µm), #2 (700-1000 µm), #3 (500-800 µm), and #4 (800-500 µm), were molded by selective laser melting technique, and the scaffolds were tested by scanning electronic microscope, uniaxial-compression tests, and Young's modulus tests; they were compared with same size pig femoral bone scaffolds. RESULTS: Under uniaxial-compression tests, equivalent stress of tantalum scaffold was 411 ± 1.43 MPa, which was significantly larger than the titanium scaffolds (P < 0.05). Young's modulus of tantalum scaffold was 2.61 ± 0.02 GPa, which was only half of that of titanium scaffold. The stress-strain curves of tantalum scaffolds were more similar to pig bone scaffolds than titanium scaffolds. CONCLUSION: 3D printed tantalum scaffolds with varying pore diameters are more similar to actual bone scaffolds compared with titanium scaffolds in biomechanical properties.

Transforming Growth Factor-ß3/Recombinant Human-like Collagen/Chitosan Freeze-Dried Sponge Primed With Human Periodontal Ligament Stem Cells Promotes Bone Regeneration in Calvarial Defect Rats.

Patients with a skull defect are at risk of developing cerebrospinal fluid leakage and ascending bacterial meningitis at >10% per year. However, treatment with stem cells has brought great hope to large-area cranial defects. Having found that transforming growth factor (TGF)-ß3 can promote the osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs), we designed a hybrid TGF-ß3/recombinant human-like collagen recombinant human collagen/chitosan (CS) freeze-dried sponge (TRFS) loading hPDLSCs (TRFS-h) to repair skull defects in rats. CFS with 2% CS was selected based on the swelling degree, water absorption, and moisture retention. The CS freeze-dried sponge (CFS) formed a porous three-dimensional structure, as observed by scanning electron microscopy. In addition, cytotoxicity experiments and calcein-AM/PI staining showed that TRFS had a good cellular compatibility and could be degraded completely at 90 days in the implantation site. Furthermore, bone healing was evaluated using micro-computed tomography in rat skull defect models. The bone volume and bone volume fraction were higher in TRFS loaded with hPDLSCs (TRFS-h) group than in the controls (p < 0.01, vs. CFS or TRFS alone). The immunohistochemical results indicated that the expression of Runx2, BMP-2, and collagen-1 (COL Ⅰ) in cells surrounding bone defects in the experimental group was higher than those in the other groups (p < 0.01, vs. CFS or TRFS alone). Taken together, hPDLSCs could proliferate and undergo osteogenic differentiation in TRFS (p < 0.05), and TRFS-h accelerated bone repair in calvarial defect rats. Our research revealed that hPDLSCs could function as seeded cells for skull injury, and their osteogenic differentiation could be accelerated by TGF-ß3. This represents an effective therapeutic strategy for restoring traumatic defects of the skull.

Diabetes health management strategy based on internet plus graded diagnosis and treatment strategy.

BACKGROUND: Type 2 diabetes (T2D) is a widespread chronic disease with high rates of morbidity and mortality worldwide. Managing risk factors can effectively prevent acute and chronic complications, improve quality of life, and reduce mortality. Therefore, implementation of a diabetes health management strategy is urgently needed. Emerging medical technologies have strengthened communication between patients and clinicians. The establishment and improvement of the graded diagnosis and treatment system has promoted the prevention, treatment, and management of diabetes. METHODS: A total of 300 patients diagnosed with T2D in the Health Management Center at the Affiliated Hospital of Nantong University were randomly divided into two groups: an internet plus graded diagnosis and treatment strategy group, and a control group. After 6 months, the physiological parameters, management indices, and complications were compared between the groups. RESULTS: Physiological indicators, such as body mass index (BMI), waist circumference, triglycerides, low-density lipoprotein, systolic and diastolic blood pressure, and blood glucose were significantly alleviated in the Internet plus graded diagnosis and treatment strategy group. Management indicators (such as blood glucose monitoring compliance rate, diet control compliance rate, and exercise compliance rate) also improved substantially. The incidence of hypoglycemia was notably increased compared to the control group. CONCLUSIONS: The new health management strategy for diabetes can improve lifestyle, ameliorate physiological indicators, reduce the complication rate, and form a virtuous cycle. This provides a positive impact on the entire life-cycle health management of diabetes, and is worthy of further promotion.

The correlations between the serum expression of miR-206 and the severity and prognosis of sepsis.

BACKGROUND: An accurate assessment of the severity and prognosis of sepsis, especially septic shock, is vital for the tailored treatment of this condition. miRNA participates in the inflammatory response and cell apoptosis and regulates inflammation-related signaling pathways. Immune disorders often accompany sepsis. Since serum miRNA expression is superior to traditional biological markers in terms of sensitivity and specificity, its role in the assessment of sepsis has increasingly been recognized. METHODS: Serum miRNAs were extracted from septic patients and healthy individuals by using the ultracentrifugation method. The differential expressions of miRNAs in the serum samples were detected by high-throughput sequencing technology. The differentially expressed miRNAs between the two groups were analyzed by bioinformatics. The quantitative polymerase chain reaction real-time polymerase chain reaction (qRT-PCR) was used to amplify the sample size to verify the results and to screen the highly-expressed miR206 in septic patients. Subsequently, serum samples were collected from 63 septic patients, and 30 patients with septic shock and qRT-PCR were performed to analyze the expression of miR-206. These 93 patients were divided into the miR-206 low-expression group and miR-206 high-expression group according to miR206 expression level. The potential correlations between the miR-206 expression and the clinical data were analyzed by using SPSS 25.0. RESULTS: Serum miRNA expression significantly differed between septic patients and healthy individuals. High-throughput sequencing results showed that, compared with those in healthy individuals, 29 miRNA molecules were down-regulated, and 25 molecules were up-regulated in the serum samples of septic patients. qRT-PCR identified the significantly up-regulated miR-206 in septic patients. qRT-PCR also showed significantly higher miR-206 expression levels in patients with septic shock than in septic patients. Furthermore, we observed a significantly longer prothrombin time and activated partial thromboplastin time, and significantly higher SOFA score, APACHE-II score, and in-hospital mortality rate. miR-206 was positively correlated with SOFA sore and APACHE-II score. CONCLUSIONS: Serum miR-206 expression is positively correlated with the severity and prognosis of sepsis. Thus, it may be a potential biomarker for assessing the severity and prognosis of sepsis, although the specific mechanism warrants further investigations.

Hybrid Freeze-Dried Dressings Composed of Epidermal Growth Factor and Recombinant Human-Like Collagen Enhance Cutaneous Wound Healing in Rats.

Epidermal growth factor (EGF) is important for promoting skin repair and remodeling. Native collagen is also widely used as a scaffold for skin tissue engineering. The limitations of EGF include easy decomposition or inactivation, whereas native collagen is immunogenic and has poor solubility. Therefore, we constructed a freeze-dried dressing based on the recombinant human-like collagen (RHC) to act as a carrier for EGF (RHC/EGF freeze-dried dressing) and promote skin wound closure. Here, the freeze-dried dressing that combined EGF and RHC significantly enhanced the proliferation, adhesion, and spreading of NIH/3T3 fibroblasts and migration of HaCaT keratinocytes at the wound site. The physicochemical characteristics of the RHC/EGF freeze-dried dressing investigated using scanning electron microscopy, Fourier transform infrared (FTIR) spectroscopy, and differential scanning calorimetry revealed that it was a loose and porous cake that redissolved quickly. The molecular mechanisms involved in cell proliferation and angiogenesis were also assessed. The expression levels of the markers Ki-67, proliferating cell nuclear antigen, vascular endothelial growth factor, and cluster of differentiation 31 were significantly increased after treatment with the RHC/EGF freeze-dried dressing (P < 0.01, vs. RHC or EGF alone). This increase indicated that the RHC/EGF freeze-dried dressing significantly accelerated wound closure, re-epithelialization, and the orderly arrangement and deposition of collagen in the Sprague-Dawley rats with full-thickness skin defects. This work describes a significant step toward the development of wound environments conducive to healing, and the RHC/EGF freeze-dried dressing is a potential therapeutic strategy in wound management.

Integrated Analysis of Transcriptomic Data Reveals Key Anti-CKD Targets and Anti-ESRD Targets.

BACKGROUND: Chronic kidney disease (CKD) is a kidney disease in which there is gradual loss of kidney function over time and end-stage renal disease (ESRD) is the final stage of CKD. Both CKD and ESRD are worldwide health problems with a high economic cost to health systems. However, the molecular mechanisms of the development of CKD and ESRD remain poorly understood. This study aimed to systematically elucidate the molecular mechanisms of the development of CKD and ESRD. METHODS: Transcriptome data of CKD and ESRD were downloaded from the NCBI-GEO database. Differentially expressed genes between cases and controls (chronic kidney disease patients vs. controls, end-stage renal disease patients vs. controls) were calculated using the empirical Bayes algorithm. Gene set enrichment analysis (GSEA) was used for analyzing the KEGG pathway difference between cases and controls. Furthermore, CKD and ESRD target genes were obtained from the Thomson Reuters Integrity database. Tissue-specific gene interaction network analysis was performed using the GIANT web server. RESULTS: There were multiple damaged pathways in ESRD but only a few pathways were disturbed in CKD. Furthermore, we identified 9 dysregulated anti-ESRD genes but no dysregulated anti-CKD genes. Network analysis revealed that the NF-kB signaling pathway was essential for ESRD. CONCLUSIONS: This study revealed several crucial anti-ESRD genes that are involved in the regulation of the NF-kB signaling pathway. This information may be helpful for the treatment of ESRD.

Activin B signaling may promote the conversion of normal fibroblasts to scar fibroblasts.

This study is to explore the molecular mechanism of benign bile duct hypertrophic scar formation.Differential proteins between the normal fibroblast (NFB) and scar fibroblast (SCFB) were screened by protein chip assay, and analyzed by pathway-enrichment analysis and function-enrichment analysis. The differential proteins were further tested by ELISA. SiRNA-Act B was transfected to SCFB to down-regulate the expression of Act B. NFB was incubated with rh-Act B. The cell apoptosis and cell cycle were determined by flow cytometry. The expression of Act B, Smad2/3, transforming growth factor-ß1 (TGF-ß1), endothelin-1 (ET-1), thrombospondin-1 (Tsp-1), and Oncostatin M (OSM) were detected by Western blot.A total of 37 differential proteins were identified in SCFBs by microarray (P < .05), including 27 up-regulated proteins and 10 down-regulated proteins (P < .05). Their function were associated with Activin signaling, synthesis and degradation of extracellular matrix, formation and activation of cytokine, inflammatory reaction, immunoreaction, tissue damage reaction, cell cycle, migration, apoptosis, and secretion, etc. ELISA results showed that the expression of Act B, TGF-ß1, ET-1 were higher in SCFBs, while the expression of Tsp-1 and OSM were lower in SCFBs (P < .05). After interfered by siRNA-Act B, the expression of Act B mRNA decreased (P < .05). The percentage of early apoptosis increased (P < .05). The expression of Act B, Smad2/3, TGF-ß1 were decreased and Tsp-1, OSM were increased (P < .05). After treatment with rh-Act B, the percentage of G0/G1 phase of NFBs was decreased and that of S phase was increased without significance (P > .05). The expression of Act B, Smad2/3, TGF-ß1 were increased (P < .05) and Tsp-1, OSM were decreased (P < .01).There are differentially expressed proteins between SCFBs and NFBs. Activin B signal plays an important role in the process of NFB transforming to SCFB, and TGF-ß1, Smad2/3, Tsp-1, and OSM are important participants.

A Mansonone Derivative Coupled with Monoclonal Antibody 4D5-Modified Chitosan Inhibit AKR1C3 to Treat Castration-Resistant Prostate Cancer.

PURPOSE: Aldo-ketoreductase (AKR) 1C3 is crucial for testosterone synthesis. Abnormally high expression/activity of AKR1C3 can promote castration-resistant prostate cancer (CRPC). A mansonone derivative and AKR1C3 inhibitor, 6e, was combined with 4D5 (extracellular fragment of the monoclonal antibody of human epidermal growth factor receptor-2)-modified chitosan to achieve a nanodrug-delivery system (CS-4D5/6e) to treat CRPC. MATERIALS AND METHODS: Morphologies/properties of CS-4D5/6e were characterized by atomic force microscopy, zeta-potential analysis, and Fourier transform-infrared spectroscopy. CS-4D5/6e uptake was measured by immunofluorescence under confocal laser scanning microscopy. Testosterone in LNCaP cells overexpressing human AKR1C3 (LNCaP-AKR1C3) and cell lysates was measured to reflect AKR1C3 activity. Androgen receptor (AR) and prostate-specific antigen (PSA) expression was measured by Western blotting. CS-4D5/6e-based inhibition of AKR1C3 was evaluated in tumor-xenografted mice. RESULTS: CS-4D5/6e was oblate, with a particle size of 200-300 nm and thickness of 1-5 nm. Zeta potential was 1.39±0.248 mV. 6e content in CS-4D5/6e was 7.3±1.4% and was 18±3.6% for 4D5. 6e and CS-4D5/6e inhibited testosterone production significantly in a concentration-dependent manner in LNCaP-AKR1C3 cells, and a decrease in expression of AKR1C3, PSA, and AR was noted. Half-maximal inhibitory concentration of CS-4D5/6e on LNCaP-AKR1C3 cells was significantly lower than that in LNCaP cells (P<0.05). CS-4D5/6e significantly reduced growth of 22Rv1 tumor xenografts by 57.00% compared with that in the vehicle group (P<0.01). CONCLUSION: We demonstrated the antineoplastic activity of a potent AKR1C3 inhibitor (6e) and its nanodrug-delivery system (CS-4D5/6e). First, CS-4D5/6e targeted HER2-positive CRPC cells. Second, it transferred 6e (an AKR1C3 inhibitor) to achieve a reduction in intratumoral testosterone production. Compared with 6e, CS-4D5/6e showed lower systemic toxicity. CS-4D5/6e inhibited tumor growth effectively in mice implanted with tumor xenografts by downregulating testosterone production mediated by intratumoral AKR1C3. These results showed a promising strategy for treatment of the CRPC that develops invariably in prostate-cancer patients.

The Curcumin Derivative, H10, Suppresses Hormone-Dependent Prostate Cancer by Inhibiting 17ß-Hydroxysteroid Dehydrogenase Type 3.

The 17ß-hydroxysteroid dehydrogenase type 3 (17ß-HSD3) enzyme is a potential therapeutic target for hormone-dependent prostate cancer, as it is the key enzyme in the last step of testosterone (T) biosynthesis. A curcumin analog, H10, was optimized for inhibiting T production in LC540 cells that stably overexpressed 17ß-HSD3 enzyme (LC540 [17ß-HSD3]) (P < 0.01), without affecting progesterone (P) synthesis. H10 downregulated the production of T in the microsomal fraction of rat testes containing the 17ß-HSD3 enzyme from 100 to 78.41 ± 7.41%, 51.86 ± 10.03%, and 45.14 ± 8.49% at doses of 10, 20, and 40 µM, respectively. There were no significant differences among the groups with respect to the protein expression levels of 17ß-HSD3, 3ßHSD1, CYP17a1, CYP11a1, and STAR, which participate in 17ß-HSD3-mediated conversion of androgens to T (P > 0.05). This indicated that H10 only inhibited the enzymatic activity of 17ß-HSD3 in vitro. Furthermore, H10 inhibited the adione-stimulated growth of xenografts established from LNCaP cells in nude mice in vivo. We conclude that H10 could serve as an effective inhibitor of 17ß-HSD3, which in turn would inhibit the biosynthesis of androgens and progression of prostate cancer.

Bioinformatics Analysis Identifies Protein Tyrosine Kinase 7 (PTK7) as a Potential Prognostic and Therapeutic Biomarker in Stages I to IV Hepatocellular Carcinoma.

BACKGROUND Worldwide, hepatocellular carcinoma (HCC) accounts for 80-90% of all cases of primary liver cancer, and is one of the ten most common malignancies. This study used bioinformatics analysis to identify genes associated with patient outcome in stages I-IV HCC and the gene pathways that distinguished between normal liver and liver cells and HCC and human HCC cell lines. MATERIAL AND METHODS Target genes were defined as those that had marketed drugs or drugs under development targeting a specific gene and acquired from the Clarivate Analytics Integrity Database. Differential expression gene analysis, co-expression network analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis, survival analysis and receiver operating characteristic (ROC) curve analysis were used to explore the similarities and differences in gene expression profiles, functional associations, and survival in stage I-IV HCC. Normal liver cells (HL-7702) and HCC cell lines (HepaRG, HepG2, SK-Hep1, and Huh7) were studied using Western blot and quantitative reverse transcription PCR (RT-qPCR). RESULTS Hierarchical gene clustering identified target genes that distinguished between HCC and normal liver tissue. For stages I-IV HCC, there were seven commonly upregulated target genes EPHB1, LTK, NTRK2, PTK7, TBK1, TIE1, and TLR3, which were mainly involved in immune and signaling transduction pathways. PTK7 was highly expressed in stage I-IV HCC and was an independent prognostic marker for reduced overall survival (OS). CONCLUSIONS Bioinformatics analysis, combined with patient survival analysis, identified PTK7 gene expression as a potential therapeutic target and prognostic biomarker for all stages of HCC.

IL-21 reinvigorates exhausted natural killer cells in patients with HBV-associated hepatocellular carcinoma in STAT1-depedent pathway.

Hepatocellular carcinoma (HCC) is the most common liver malignancy with dismal prognosis and limited treatment options. Natural killer (NK) cells are critical components of antitumor immunity due to their capacity to eliminate MHC class I-deficient cells. To evaluate the function of NK cells in HCC patients, circulating CD3-CD56+ NK cells were collected from HBV-associated HCC patients and healthy control individuals. Compared to NK cells from healthy controls, NK cells from HCC patients presented functional impairment, characterized by significantly reduced cytotoxicity, degranulation, and cytokine production. Exogenous IL-21 could reinvigorate NK cells from HCC patients, resulting in significantly increased levels of cytotoxicity, degranulation, and cytokine expression. However, IL-21-treated NK cells from HCC patients still presented lower response than IL-21-treated NK cells from healthy controls. IL-21 resulted in increased phosphorylation of both STAT1 and STAT3 in NK cells. Inhibition of STAT1, but not STAT3, significantly reduced IL-21-mediated reinvigoration of NK function. Together, this study demonstrated that NK cells in HBV-associated HCC patients presented functional impairments that could be reverted by IL-21 in a STAT1-mediated mechanism.

Improving the protective effects of aFGF for peripheral nerve injury repair using sulfated chitooligosaccharides.

Injury to the peripheral nerves can result in temporary or life-long neuronal dysfunction and subsequent economic or social disability. Acidic fibroblast growth factor (aFGF) promotes the growth and survival of neurons and is a possible treatment for peripheral nerve injury. Yet, the actual therapeutic utility of aFGF is limited by its short half-life and instability in vivo. In the present study, we prepared sulfated chitooligosaccharides (SCOS), which have heparin-like properties, to improve the bioactivity of aFGF. We investigated the protective effects of SCOS with or without aFGF on RSC96 cells exposed to Na2S2O4 hypoxia/reoxygenation injury. Cell viability was measured by MTT assay and cytotoxicity induced by Na2S2O4 was assessed by lactate dehydrogenase (LDH) release into the culture medium. Pretreatment with aFGF and SCOS dramatically decreased LDH release after injury compared to pretreatment with aFGF or SCOS alone. We subsequently prepared an aFGF/SCOS thermo-sensitive hydrogel with poloxamer and examined its effects in vivo. Paw withdrawal thresholds and thermal withdrawal latencies were measured in rats with sciatic nerve injury. Local injection of the aFGF/SCOS hydrogels (aFGF: 40, 80 µg/kg) increased the efficiency of sciatic nerve repair compared to aFGF (80 µg/kg) hydrogel alone. Especially aFGF/SCOS thermo-sensitive hydrogel decreased paw withdrawal thresholds from 117.75 ± 8.38 (g, 4 d) to 65.74 ± 3.39 (g, 10 d), but aFGF alone group were 140.58 ± 27.54 (g, 4 d) to 89.12 ± 5.60 (g, 10 d) (aFGF dose was 80 µg/kg, P < 0.05, n = 8). The thermal withdrawal latencies decreased from 11.61 ± 2.26 (s, 4 d) to 2.37 ±0.67 (s, 10 d). However, aFGF alone group were from 17.69 ± 1.47 (s, 4 d) to 4.65 ± 1.73 (s, 10 d) (P < 0.05, n = 8). Furthermore, the aFGF/SCOS hydrogels also exhibited good biocompatibility in mice. In summary, SCOS improved the protective effects of aFGF in RSC96 cells injured with Na2S2O4 and increased the efficiency of nerve repair and recovery of function in rats with sciatic nerve injury. These findings pave an avenue for the development of novel prophylactic and therapeutic strategies for peripheral nerve injury.

Pilot-scale expression, purification, and bioactivity of recombinant human TGF-ß3 from Escherichia coli.

TGF-ß3, a subtype of transforming growth factor-ß (TGF-ß), is essential to various biological processes, including endoderm development, organogenesis, epithelial hyperplasia, synthesis of extracellular matrix, and immune response. Essentially, TGF-ß3 engages the TGF-ß1/Smad signaling pathway to stimulate mesenchymal lineage cells, inhibit epithelial or neuroectodermal lineage cells, and regulate repair, remodeling, and potential scarring after cutaneous wounding. We have now expressed recombinant human TGF-ß3 in Escherichia coli Origami B (DE3), with yield 300 ±â€¯17 mg/L monomeric protein at pilot scale. Identity was confirmed by western blot and HPLC-based peptide mapping. After purification and refolding, dimeric proteins were found to induce chondro-related genes in adipose-derived stem cells, and to suppress scarring in injured rabbit ears. Thus, the recombinant protein has excellent potential for medical applications.

CXCR5+CD8+ T cells could induce the death of tumor cells in HBV-related hepatocellular carcinoma.

The follicular CXCR5+CD8+ T cells have recently emerged as a critical cell type in mediating peripheral tolerance as well as antiviral immune responses during chronic infections. In this study, we investigated the function of CXCR5+CD8+ T cells in HBV-related hepatocellular carcinoma patients. Compared to CXCR5-CD8+ T cells, CXCR5+CD8+ T cells presented elevated PD-1 expression but reduced Tim-3 and CTLA-4 expression. Upon anti-CD3/CD28 stimulation, CXCR5+CD8+ T cells demonstrated higher proliferation potency than CXCR5-CD8+ T cells, especially after PD-1 blockade. CXCR5+CD8+ T cells also demonstrated significantly higher granzyme B synthesis and release, as well as higher level of degranulation. Tumor cells were more readily eliminated by CXCR5+CD8+ T cells than by CXCR5-CD8+ T cells. Interestingly, we found that B cells were more resistant to CXCR5+CD8+ T cell-mediated killing than tumor cells, possibly through IL-10-mediated protection. In addition, the CXCR5+CD8+ T cell-mediated cytotoxic effects on tumor cells could be significantly enhanced by PD-L1 blockade. Together, we presented that in patients with in HBV-related hepatocellular carcinoma, CXCR5+CD8+ T cells could mediate tumor cell death more potently than the CXCR5-CD8+ T cells in vitro while the autologous B cells were protected.

Role of Small Interfering RNA Silencing Protein Kinase C-α Gene on the Occurrence of Ultrafiltration Failure in Peritoneal Dialysis Rats.

This study aims to explore the effect of PKC-α gene silencing on the occurrence of ultrafiltration failure (UFF) in peritoneal dialysis (PD) rats. Forty-eight male SD rats were collected to establish 5/6 renal resection uremic and uremic PD rats models. Rats were assigned into control, sham operation, uremia, PD-2 W (peritoneal dialysis for 2 weeks), PD-4 W (peritoneal dialysis for 4 weeks), negative control (NC) (peritoneal dialysis for 4 weeks, and injected 0.1 mg/kg blank plasmid into abdominal cavity) and PKC-α siRNA (peritoneal dialysis for 4 weeks, and injected 0.1 mg/kg PKC-α siRNA into abdominal cavity) groups. CD34 staining was performed to determine microvessel density (MVD) for peritoneal tissues. The mRNA and protein expression of PKC-α in peritoneal tissue were detected by qRT-PCR and Western blot. Compared with the control group, MVD, the mRNA and protein expression of PKC-α were significantly increased in rats of the uremia, PD-2 W, PD-4 W, NC, and PKC-α siRNA groups. Compared with the uremia group, MVD, the mRNA and protein expression of PKC-α were increased, the changes observed in the PD-4 W and NC groups were better obvious than in the PD-2 W group. In comparison with the PD-4 W and NC groups, MVD, the mRNA and protein expression of PKC-α in rats were decreased in the PKC-α siRNA group. PKC-α gene has a high expression in uremic PD rats, and PKC-α gene silencing is able to increase UF while decrease MVD and glucose transport in peritoneal tissues thus reversing UFF in PD rats. J. Cell. Biochem. 118: 4607-4616, 2017. © 2017 Wiley Periodicals, Inc.

Discussion on software aging management of nuclear power plant safety digital control system.

Managing the aging of digital control systems ensures that nuclear power plant systems are in adequate safety margins during their life cycles. Software is a core component in the execution of control logic and differs between digital and analog control systems. The hardware aging management for the digital control system is similar to that for the analog system, which has matured over decades of study. However, software aging management is still in the exploratory stage. Software aging evaluation is critical given the higher reliability and safety requirements of nuclear power plants. To ensure effective inputs for reliability assessment, this paper provides the required software aging information during the life cycle. Moreover, the software aging management scheme for safety digital control system is proposed on the basis of collected aging information.

MicroRNA-200b inhibits the proliferation of hepatocellular carcinoma by targeting DNA methyltransferase 3a.

Aberrant microRNA (miRNA or miR) expression has been reported to contribute to the pathogenesis of hepatocellular carcinoma (HCC). However, the involvement of specific miRNAs in HCC remains to be elucidated. The present study aimed to investigate the potential role of miR-200b and the mechanism underlying its function in hepatocarcinogenesis. The results of the present study demonstrated that the expression levels of miR­200b were significantly reduced in HCC tissue samples, as compared with normal liver (NL) and para­tumorous (PT) tissue samples. The results also revealed that miR­200b expression levels in HepG2 cells were significantly decreased compared with those in L02 cells. In addition, western blotting and reverse transcription­quantitative polymerase chain reaction demonstrated that the expression levels of DNA methyltransferase 3a (DNMT3a), a possible target gene for miR­200b, were significantly higher in HCC tissue samples, as compared with those in NL and PT tissue samples. Furthermore, the data suggested that DNMT3a was a direct target gene of miR­200b. Upregulated miR­200b expression in HepG2 cells led to a decrease in DNMT3a expression levels, and an inhibition of cell proliferation. These results suggested that miR­200b has an important role in hepatocarcinogenesis and acts by downregulating DNMT3a expression. Thus, miR-200b may be a promising target for HCC treatment.

[miR-144 regulates BCG- and rapamycin-induced autophagy by targeting Atg4a in RAW264.7 cells].

OBJECTIVE: To study the effect of Bacilli Calmette-Guerin (BCG) on miRNAs (miR-21, miR-181a, miR-155 and miR-144) in RAW264.7 cells, and with miR-144 as an example, to verify the relationship of miR-144 and Atg4a, an autophagy-related gene, and study the mechanism underlying the regulatory effect of miR-144 on autophagy in the process of Mycobacterium tuberculosis infection. METHODS: RAW264.7 cells were treated respectively with starvation (12 hours), 50 ng/mL rapamycin (2 hours) and 10 nmol/L 3-methyl adenine (3-MA, 12 hours), and then stimulated with BCG for 12, 24 or 48 hours. The macrophages were collected for total RNA extraction. The expression levels of miR-21, miR-181a, miR-155 and miR-144 were detected by real-time quantitative PCR (qRT-PCR). Thereafter, we constructed the recombinant plasmids pMIR-Report-Atg4a and pMIR-Report-Atg4a mut. The targeting effect of miR-144 on Atg4a gene was verified by the dual-luciferase reporter assay system, Western blotting and qRT-PCR. RESULTS: After BCG stimulated RAW264.7 cells, the expression levels of miR-21, miR-155 and miR-144 were up-regulated, and miR-181a expression was down-regulated. The expressions of miR-21, miR-144, miR-155 and miR-181a increased 64 times, 52 times, 14 times and 1 times in the rapamycin group, respectively; the expressions of miR-21, miR-144, miR-155 and miR-181a decreased 1.22 times, 1.05 times, 1.54 times and 12.5 times in the 3-MA group, respectively. The dual-luciferase reporter assay system and Western blotting demonstrated that miR-144 could suppress Atg4a expression by targeting the specific 3'-untranslated region (3'UTR) sequence of Atg4a gene. CONCLUSION: miR-144 can directly inhibit the autophagy-related gene Atg4a expression and participate in the regulation of autophagy process in Mycobacterium tuberculosis infection.

Comparison of patterns and knowledge of benefits and warnings of fish consumption between parents and children.

We examined generational differences in fish consumption and knowledge of benefits/warnings of fish consumption among parents and children. This cross-sectional study gathered self-administered questionnaire data, including demographics, fish consumption behavior (including specific fish species) and knowledge of fish consumption warnings and benefits. Fish were later grouped into four categories by potential mercury contamination. Descriptive statistics were conducted for all variables comparing all adults and children. Benefit/risk knowledge variables were also descriptively analyzed among parent-child pairs only. Multivariate Poisson regression was conducted on pairs to assess risk factors for children eating higher mercury fish. 421 adults and 207 children (171 adult-child pairs) participated (family response rate: 71%). Slightly more adults (97.6%) ate fish in the last year than children (92.3%); however, there was no difference between consumption of fish by category of potential mercury contamination. Both adults (44%) and children (45%) ate high-mercury fish. In 71% of parent-child pairs, both the parent and the child knew of benefits of consuming fish; only 31% knew of warnings. Parental consumption of high or moderately-high-mercury fish was related to the child's consumption of fish in the same category. Parents and children need additional education to make better choices about fish consumption. Education should target the family and include specifics about benefits and risks.