Dual Role of IGF2BP2 in Osteoimmunomodulation during Periodontitis.

Periodontitis is a complex disease characterized by distinct inflammatory stages, with a peak of inflammation in the early phase and less prominent inflammation in the advanced phase. The insulin-like growth factor 2-binding proteins 2 (IGF2BP2) has recently been identified as a new m6A reader that protects m6A-modified messenger RNAs (mRNAs) from decay, thus participating in multiple biological processes. However, its role in periodontitis remains unexplored. Here, we investigated the role of IGF2BP2 in inflammation and osteoclast differentiation using a ligature-induced periodontitis model. Our findings revealed that IGF2BP2 responded to bacterial-induced inflammatory stimuli and exhibited differential expression patterns in early and advanced periodontitis stages, suggesting its dual role in regulating this disease. Depletion of Igf2bp2 contributed to increased release of inflammatory cytokines, thereby exacerbating periodontitis after 3 d of ligature while suppressing osteoclast differentiation and ameliorating periodontitis after 14 d of ligature. Mechanistically, we demonstrated that IGF2BP2 directly interacted with Cd5l and Cd36 mRNA via RNA immunoprecipitation assay. Overexpression of CD36 or recombinant CD5L rescued the osteoclast differentiation ability of Igf2bp2-null cells upon lipopolysaccharide stimulus, and thus the downregulation of Cd36 and Cd5l effectively reversed periodontitis in the advanced stage. Altogether, this study deepens our understanding of the potential mechanistic link among the dysregulated m6A reader IGF2BP2, immunomodulation, and osteoclastogenesis during different stages of periodontitis.

The effect of microRNA-663b in the inhibition of interleukin-1-induced nucleus pulposus cell apoptosis and inflammatory response.

The aim of this study was to explore the role and pathological mechanism of microRNA-663b in interleukin-1beta (IL-1ß)-induced inflammation and apoptosis of nucleus pulposus cells. First, the best concentration and time to construct the nucleus pulposus cell inflammation model was screen out. Overexpression or inhibition of miR-663b expression was performed by adding microRNA-663b mimic or microRNA-663b inhibitor. 293T cells were transfected according to experimental requirements. The luciferase activity of each group was detected to analyze the targeted regulation of microRNA-663b on interleukin-1 receptor (IL1R1). Compared with the mimic negative control (NC) group, the expression of inflammatory factors in the microRNA-663b overexpression group was inhibited (P<0.05), and the expression of type 2 collagen and polysaccharide protein increased (P<0.05), and the apoptosis of nucleus pulposus cells was inhibited (P<0.01), and the number of TUNEL-positive cells decreased significantly (P<0.01), and the microRNA and protein expression of IL1R1, the ratio of P-P65/P65 and phospho-nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor, alpha (P-IκBα)/nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor, alpha (IκBα) protein expression were significantly decreased (P<0.05). The expression of inflammatory factors in the miR-663b inhibitor group was significantly higher than that in the inhibitor NC group (P<0.01), and the expression of type 2 collagen and polysaccharide protein was significantly decreased (P<0.01), and the number of apoptosis cells and TUNEL staining positive cells increased (p<0.01). The expression of IL1R1 gene and protein was significantly increased (P<0.01). The ratio of P-P65/P65 and P-IκBα/IκBα protein expression increased (P<0.05). IL1R1 is a downstream target gene of microRNA-663b. MicroRNA-663b may down-regulate the expression of IL1R1 at the transcriptional level by targeting IL1R1, inhibit the inflammatory response of nucleus pulposus cells, and slow down the degeneration of nucleus pulposus cells.

[Bronchoscopic manifestations and interventional treatment of pulmonary mucormycosis].

Objective: To analyze the bronchoscopic manifestations and interventional treatment of pulmonary mucormycosis. Methods: Clinical data of patients with pulmonary mucormycosis undergoing bronchoscopy and interventional therapy in 4 tertiary general hospitals in China from May 2006 to May 2022 were retrospectively analyzed and the literature on the subject were reviewed. Results: The data of 10 patients with pathologically diagnosed pulmonary mucormycosis undergoing bronchoscopy and interventional therapy were collected, including 8 males and 2 females. The patients' age ranged from 21 to 72 (44±15) years. The underlying diseases included 6 cases of diabetes ketoacidosis, 3 cases of leukemia, 1 case after operation of lung cancer. Bronchoscopy showed that white viscous necrotic matters grew along the airway and blocked the airway in 9 cases, accompanied by airway bleeding in 3 cases, bloody secretion blocked the airway in 1 case, and bronchopulmonary cavity fistula in 2 cases. The biopsy histopathology of white necrotic matters showed that many mucor filaments were tangled together which were named mucormycelium. Among the 10 patients, 9 were treated with systemic drugs, including intravenous application of amphotericin B deoxycholate in 5 cases, intravenous application of amphotericin B liposome in 4 cases, oral posaconazole in 6 cases and intravenous injection in 1 case. Local drug therapy included aerosol inhalation of amphotericin B deoxycholate in 8 cases and local perfusion under bronchoscope in 5 cases. Bronchoscopic interventional therapy was used to remove mucormycelium in the bronchus, including cryotherapy in 8 cases, biopsy forceps in 7 cases, snare treatment in 2 cases and foreign body forceps in 2 cases. All 10 patients were clinical cured and with no death. Conclusions: Pulmonary mucormycosis is more common in immunocompromised hosts. Bronchoscopy often showed mucormycelium blocking the airway. Systemic and local drug therapy combined with bronchoscopic interventional therapy can achieve good clinical efficacy.

Nucleus pulposus cells degeneration model: a necessary way to study intervertebral disc degeneration.

The availability of an appropriate and reliable research model is helpful for researchers to understand the occurrence and development of diseases. Historically, animal models have been beneficial in the study of intervertebral disc degenerative diseases, but intervertebral disc degeneration (IDD) is a precise and complex process that needs to appear and occur in a specific tissue microenvironment, and animal degeneration models cannot fully simulate these parameters. These challenges must be overcome, especially when animal models cannot fully generalise the complex pathology of humans. In the past few years, the research on the cell disease model has made important progress, and the construction of the nucleus pulposus cell (NPC) degeneration model has become an indispensable step in studying the occurrence and development of IDD. Here, several different methods of constructing NPC degeneration models and indicators for testing the effect of modelling are introduced. The practical applications of cell models constructed by different methods are enumerated to screen and evaluate effective methods of establishing degenerative cell models and explore the mechanism of IDD.

Ncor1 Deficiency Promotes Osteoclastogenesis and Exacerbates Periodontitis.

Nuclear receptor corepressor 1 (Ncor1) has been reported to regulate different transcription factors in different biological processes, including metabolism, inflammation, and circadian rhythms. However, the role of Ncor1 in periodontitis has not been elucidated. The aims of the present study were to investigate the role of Ncor1 in experimental periodontitis and to explore the underlying mechanisms through an experimental periodontitis model in myeloid cell-specific Ncor1-deficient mice. Myeloid cell-specific Ncor1 knockout (MNKO) mice were generated, and experimental periodontitis induced by ligation using 5-0 silk sutures was established. Ncor1 flox/flox mice were used as littermate controls (LC). Histological staining and micro-computed tomography scanning were used to evaluate osteoclastogenesis and alveolar bone resorption. Flow cytometry was conducted to observe the effect of Ncor1 on myeloid cells. RNA sequencing was used to explore the differentially targeted genes in osteoclastogenesis in the absence of Ncor1. Coimmunoprecipitation (Co-IP), chromatin immunoprecipitation (ChIP) experiments, and dual luciferase assays were performed to explore the relationship between NCoR1 and the targeted gene. Alveolar bone resorption in the MNKO mice was significantly greater than that in the LC mice after periodontitis induction and osteoclastogenesis in vitro. The percentage of CD11b+ cells, particularly CD11b+ Ly6G+ neutrophils, was substantially higher in gingival tissues in the MNKO mice than in the LC mice. Results of RNA sequencing demonstrated that CCAAT enhancer binding protein α (Cebpα) was one of the most differentially expressed genes between the MNKO and LC groups. Mechanistically, Co-IP assays, ChIP experiments, and dual luciferase assays revealed that NCOR1 interacted with peroxisome proliferator-activated receptor gamma (PPARγ) and cooperated with HDAC3 to control the transcription of Cebpα. In conclusion, Ncor1 deficiency promoted osteoclast and neutrophil formation in mice with experimental periodontitis. It regulated the transcription of Cebpα via PPARγ to promote osteoclast differentiation.

Anisotropic long-range spin transport in canted antiferromagnetic orthoferrite YFeO3.

In antiferromagnets, the efficient transport of spin-waves has until now only been observed in the insulating antiferromagnet hematite, where circularly (or a superposition of pairs of linearly) polarized spin-waves diffuse over long distances. Here, we report long-distance spin-transport in the antiferromagnetic orthoferrite YFeO3, where a different transport mechanism is enabled by the combined presence of the Dzyaloshinskii-Moriya interaction and externally applied fields. The magnon decay length is shown to exceed hundreds of nanometers, in line with resonance measurements that highlight the low magnetic damping. We observe a strong anisotropy in the magnon decay lengths that we can attribute to the role of the magnon group velocity in the transport of spin-waves in antiferromagnets. This unique mode of transport identified in YFeO3 opens up the possibility of a large and technologically relevant class of materials, i.e., canted antiferromagnets, for long-distance spin transport.

Protective effect of estrogen receptors (ERα/ß) against the intervertebral disc degeneration involves activating CCN5 via the promoter.

OBJECTIVE: Due to the decrease of estrogen and estrogen receptor (ER) in postmenopausal women, they have a higher risk of intervertebral disc degeneration (IDD) than men. This study aims to explore how ERα and ERb interact with CCN5 and protect IDD. PATIENTS AND METHODS: We used Chromatin immunoprecipitation (ChIP) and Luciferase reporter assay to determine whether the ERα/b protein binds to CCN5 promoter and activates its expression. We used TNF-α to induce nucleus pulposus (NP) cell degeneration to simulate the IDD process. The change of the expression of ERα/ß and CCN5 was measured in the degenerated NP cells. To understand the function of ERα/ß in the NP cells degeneration, we upregulated the ERα/b gene expression by vector transfection or 17b-estradiol (E2) stimulation. Besides, we also used the CCN5 gene-silenced NP cells by siRNA transfection as a comparison to determine the role of CCN5. We tested the cell proliferation and principal components of the extracellular matrix (ECM) to value the degree of NP cell degeneration. RESULTS: ERα and ERß protein can bind to the same promoter regions of CCN5 and activate its expression, respectively. TNF-α degraded NP cells with a reduction of cell proliferation, collagen II, ACAN, ERα, ERß, and CCN5 expression, and increased collagen I/III, and MMP-13 expression. Upregulated ERα or ERß resulted in the maintains of CCN5 and alleviated the NP cell degeneration. Besides, 17ß-E2 supplement increased the ERα, ERß, and CCN5 expression, as well as stable NP cells phenotype. However, it was partly abolished by the silencing of CCN5. CONCLUSIONS: Upregulation of ERα and ERß protects the NP cell degeneration during IDD through the activation of CCN5 by binding to its promoter.

Ethnic Disparities in Dental Caries among Adolescents in China.

Comprehensive research on ethnic disparities in dental caries in China is limited. The aims of this cross-sectional study were to compare the levels of dental caries in adolescents between the Han ethnic group and ethnic minority groups in China and to explore the risk indicators for dental caries within ethnic subgroups. Data from the Fourth National Oral Health Survey in 2015, which covered all 31 province-level administrative divisions in mainland China, were used. The dental caries status in the permanent dentition of adolescents aged 12, 13, 14, and 15 y was measured using the decayed, missing, and filled teeth (DFMT) score, and sociodemographic characteristics and oral health-related behaviors were also collected. A total of 118,601 adolescents were included, with ethnic minority groups accounting for 13.15%. Of the Han and minority groups, the standardized prevalence of dental caries experience was 40.58% and 47.67%, and the mean DMFT scores were 0.97 and 1.28, respectively. According to the multivariate zero-inflated negative binomial regression analysis, the caries status of minorities was more severe than Han adolescents (adjusted prevalence rate ratio [PRR], 1.14; 95% confidence interval [CI], 1.10-1.18). This disparity was greater among adolescents who lived in rural areas, had mid-level economic status, and frequently consumed sugary beverages. After propensity score matchings, Uygur (PRR, 1.44; 95% CI, 1.25-1.67), Tibetan (PRR, 1.39; 95% CI, 1.3-1.48), and Yi (PRR, 1.24; 95% CI, 1.04-1.48) adolescents were significantly more likely to have caries than Han adolescents. Subgroup analyses revealed that gender, age, location of residence, economic status, region, consumption of sweet snacks and sugary beverages, and dental visit pattern were significantly associated with dental caries within ethnic minorities.

[Association of Semaphorin 3A with thrombocytopenia in systemic lupus erythematosus].

OBJECTIVE: To measure the level of serum Semaphorin 3A (Sema3A) and to analyze the relationship between serum Sema3A and systemic lupus erythematosus (SLE) with thrombocytopenia. METHODS: The concentration of serum Sema3A was detected by enzyme-linked immuno sorbent assay (ELISA) in 170 SLE patients, 50 Sjögren's syndrome (SS) patients, 19 hypersplenism (HS) patients and 150 healthy controls (HC). Based on the presence of thrombocytopenia and whether the thrombocytopenia was in remission, the SLE patients were divided into three groups: SLE with thrombocytopenia (41 cases), SLE with thrombocytopenia remission (28 cases), and SLE without thrombocytopenia (101 cases). According to whether there was thrombocytopenia, the SS patients were divided into SS with thrombocytopenia (18 cases) and SS without thrombocytopenia (32 cases). The 28 SLE patients who underwent bone marrow aspiration biopsy were divided into two groups from the aspect of whether the bone marrow hyperplasia was normal (19 cases) or low (9 cases), as well as from the aspect of whether the maturity disturbance of megakaryocyte was positive (8 cases) or negative (20 cases). The serum Sema3A levels in SLE, SS, HS with HC were compared, meanwhile, the correlation between serum Sema3A level and platelet (PLT) in the patients with different diseases analyzed. RESULTS: (1) Serum Sema3A levels in SLE were significantly lower than in HC [(3.84±2.76) µg/L vs. (6.96±2.62) µg/L, P < 0.001], serum Sema3A levels in SS were also obviously lower than in HC [(4.35±3.57) µg/L vs. (6.96±2.62) µg/L, P < 0.001], and in HS it was lower than HC at a certain extant [(5.67±2.26) µg/L vs. (6.96±2.62) µg/L, P=0.041]. (2) Serum Sema3A levels in SLE were slightly lower than in SS, but there was no significant difference [(3.84±2.76) µg/L vs. (4.35±3.57) µg/L, P=0.282]. However, when compared with HS, serum Sema3A levels in SLE were significantly lower [(3.84±2.76) µg/L vs. (5.67±2.26) µg/L, P=0.006]. (3) Serum Sema3A concentration in SLE with thrombocytopenia was significantly lower than in SLE with thrombocytopenia remission [(1.28±1.06) µg/L vs. (3.83±2.65) µg/L, P < 0.001], and in SLE patients without thrombocytopenia [(1.28±1.06) µg/L vs. (4.87±2.60) µg/L, P < 0.001]. There was no significant difference between SLE with thrombocytopenia remission and SLE without thrombocytopenia [(3.83±2.65) µg/L vs. (4.87±2.600 µg/L, P=0.123]. Serum Sema3A concentration in SLE with thrombocytopenia was slightly lower than in SS with thrombocytopenia, but there was no significant difference [(1.28±1.06) µg/L vs. (1.68±1.11) µg/L, P=0.189]. (4) Strong positive correlations were found between serum Sema3A and PLT in SLE (r=0.600, P < 0.001). Positive correlations were also found between serum Sema3A and PLT in SS (r=0.573, P < 0.001). However, there was no such correlation showed in HS patients (P=0.393). (5) There was no significant difference of serum Sema3A concentration in SLE whether the bone marrow hyperplasia was normal or low. And the same situation appeared in the patients whether the maturity disturbance of megakaryocyte was positive or negative (P>0.05). CONCLUSION: Serum Sema3A was significantly reduced in SLE patients, and it was highly correlated with the blood damage. Similar conclusions could be drawn in patients with SS. The serum level of Sema3A was generally decreasing in desmosis which merged thrombocytopenia, and was obviously positive correlated with platelet counts.

m6A methyltransferase METTL3 promotes the progression of prostate cancer via m6A-modified LEF1.

OBJECTIVE: This study aims to uncover the regulatory effects of METTL3 on promoting the progression of prostate cancer (PCa) through N6-Methyladenosine (m6A) methylation on LEF1 mRNA. PATIENTS AND METHODS: The relative levels of METTL3 and LEF1 in 48 paired PCa tissues and adjacent ones were determined by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Their correlation in PCa tissues was analyzed by Spearman correlation test. The survival of PCa patients affected by METTL3 was assessed by introducing Kaplan-Meier method. Wound closure assay was performed to evaluate the potential influence of METTL3 on migratory ability in PC-3 cells. Protein level of LEF1 in PC-3 cells with METTL3 or IGF2BP2 knockdown was examined. The activity of the Wnt pathway was tested through TOP/FOP-Flash. Furthermore, the interaction between LEF1 with METTL3 or IGF2BP2 was verified through RIP (RNA-Binding Protein Immunoprecipitation) assay. At last, the regulatory effects of METTL3/LEF1 axis on the activity of the Wnt pathway and migratory ability in PC-3 cells were determined. RESULTS: METTL3 and LEF1 were upregulated in PCa tissues, and they presented a positive correlation in PCa. A high level of METTL3 predicted poor prognosis in PCa patients. The knockdown of METTL3 suppressed the migratory ability in PC-3 cells. Meanwhile, the knockdown of METTL3 downregulated protein level of LEF1 and decreased the activity of the Wnt pathway. The results of RIP assay indicated that METTL3 methylation sites were present on LEF1 mRNA. Moreover, the silence of METTL3 decreased the enrichment abundance of LEF1 in anti-IGF2BP2. Rescue experiments demonstrated that the overexpression of LEF1 partially reversed the regulatory effects of METTL3 on the Wnt activity and migratory ability in PCa cells. CONCLUSIONS: METTL3 is upregulated in PCa tissues. METTL3 influences the activity of the Wnt pathway through m6A methylation on LEF1 mRNA, thereafter, promoting the progression of PCa.

Influence of miR-199a on rats with non-small cell lung cancer via regulating the HIF-1α/VEGF signaling pathway.

OBJECTIVE: Micro-ribonucleic acids (miRNAs) are involved in the occurrence of various cancers, and the hypoxia-inducible factor 1-α (HIF-1α) is the main regulator of cell proliferation induced by hypoxia. The relationships of miR-199a and HIF-1α expressions with non-small cell lung cancer (NSCLC) remain unclear, so they were explored in this work. MATERIALS AND METHODS: On the basis of establishing the rat model of NSCLC, the messenger RNA (mRNA) expressions of miR-199a, HIF-1α and the vascular endothelial growth factor (VEGF) were analyzed in NSCLC rats, and the correlations of miR-199a with the mRNAs of HIF-1α and VEGF and cancer staging were investigated. To further study the role of miR-199a in NSCLC cell proliferation via the HIF-1α/VEGF signaling pathway, NSCLC cells were treated with the signaling pathway inhibitor and transfected with miR-199a mimics, respectively. Also,  the roles of the inhibitor PX-478 and miR-199a mimics in the expressions of miR-199a, HIF-1α, and VEGF proteins, as well as their influences on cell proliferation ability were detected. RESULTS: In NSCLC rats, the expression of miR-199a was substantially decreased (p<0.01), but the expressions of HIF-1α and VEGF were notably raised (p<0.01). MiR-199a was negatively correlated with the expression of VEGF. As cancer developed, the expression of miR-199a was gradually lowered, but the expressions of HIF-1α and VEGF were gradually increased. Both HIF-1α/VEGF signaling pathway inhibitor PX-478 and miR-199a mimics significantly reduced the expressions of HIF-1α and VEGF proteins (p<0.01) and suppressed the cell proliferation activity. CONCLUSIONS: MiR-199a prevents the proliferation of NSCLC cells via the targeted down-regulation of the HIF-1α/VEGF signaling pathway.

Characterization of the complete mitochondrial genome sequence of Tracheophilus cymbius (Digenea), the first representative from the family Cyclocoelidae.

Tracheophilus cymbius (Trematoda: Cyclocoelidae) is a common tracheal fluke of waterfowl, causing serious loss in the poultry industry. However, taxonomic identification of T. cymbius remains controversial and confused. Mitochondrial (mt) genomes can provide genetic markers for the identification of closely related species. We determined the mt genome of T. cymbius and reconstructed phylogenies with other trematodes. The T. cymbius mt genome is 13,760 bp in size, and contains 12 protein-coding genes (cox 1-3, nad 1-6, nad 4L, cyt b and atp 6), 22 transfer RNA (tRNA) genes, two ribosomal RNA genes and one non-coding region. All are transcribed in the same direction. The A + T content is 62.82%. ATG and TAG are the most common initiation and termination codons, respectively. Phylogenetic analyses of concatenated nucleotide sequences show T. cymbius grouping in suborder Echinostomata, and clustering together, with high statistical support, as a sister taxon with Echinochasmus japonicus (Echinochasmidae), the two forming a distinct branch rooted to the ancestor of all Echinostomatidae and Fasciolidae species. This is the first report of the T. cymbius mt genome, and the first reported mt genome within the family Cyclocoelidae. These data will provide a significant resource of molecular markers for studying the taxonomy, population genetics and systematics of trematodes.

Knockdown of lncRNA ZFAS1 inhibits progression of nasopharyngeal carcinoma by sponging miR-135a.

Nasopharyngeal carcinoma (NPC) is one common head and neck malignancy with leading cause of cancer-related death. Long noncoding RNAs (lncRNAs) have been reported to play essential roles in progression, prognosis and treatment of NPC. However, the exact role of lncRNA zinc finger antisense 1 (ZFAS1) in NPC progression and its potential mechanism remain largely unknown.The expressions of ZFAS1 and microRNA-135a (miR-135a) were measured in NPC tissues or cells by quantitative real-time polymerase chain reaction (qRT-PCR). The interaction between ZFAS1 and miR-135a was explored by luciferase reporter assay and RNA immunoprecipitation (RIP). Cell proliferation, apoptosis, migration and invasion were analyzed by 3-(4,5-dimethyl-2-thiazolyl)-2,5 -diphenyl-2-H-tetrazolium bromide (MTT) assay, flow cytometry or trans-well assay, respectively. Our data showed the expression of ZFAS1 was up-regulated and miR-135a was down-regulated in NPC tissues and cells. miR-135a was bound to ZFAS1 in NPC cells. Moreover, knockdown of ZFAS1 or addition of miR-135a inhibited cell proliferation, migration and invasion but promoted apoptosis in NPC cells. Besides, down-regulation of miR-135a reversed abrogation of ZFAS1-mediated inhibition of proliferation, migration and invasion and increase of apoptosis in NPC cells. Our data suggested Inhibition of ZFAS1 protected against proliferation, migration and invasion but contributed to apoptosis by sponging miR-135a in NPC cells, providing a novel avenue for NPC treatment.

Fatigue device driven by a three degree of freedom tripodal piezoelectric actuator.

A miniature piezoelectric-driven fatigue device with three degrees of freedom is developed. The device integrates two fatigue testing functions, including uniaxial tensile fatigue and tensile-bending combined loading modes. The synchronous tensile-bending loading principle is described, which is applicable for calculating the vector displacements along two orthogonal directions and investigating the anisotropic fatigue properties. Regarding the combined loading mode, maximum load/displacement amplitudes for tensile and bending vector components that could be achieved are 16.9 N/22.8 µm and 3.3 N/5.6 µm, respectively. Based on tensile and tensile-bending combined fatigue loading modes, the displacement responses and fatigue lives at loading frequencies ranging from 1 Hz to 100 Hz are valuated experimentally to indicate the validation.

Efficacy of continuous renal replacement on acute renal injury developed in severe sepsis.

Sepsis is a systemic inflammatory response that can further develop into severe sepsis (septic shock), which eventually leads to multiple organ dysfunction syndrome (MODS). This study aimed to assess the effects of continuous renal replacement therapy (CRRT) on acute renal injury caused by severe sepsis by monitoring biochemical parameters. A total of 60 patients with septic shock and acute kidney injury were included. The control group (30 cases) was treated with routine treatment and intermittent renal replacement therapy (IRRT). The experimental group (30 cases) was treated with routine treatment and continuous renal replacement therapy CRRT. The changes in inflammation and biochemical indexes and APACHE- II score were evaluated before the treatment and 1, 3, and 7days after the treatment. The inflammatory markers (neutrophil percentage, C-reactive protein (CRP) and procalcitonin (PCT) levels) in the experimental group decreased significantly after treatment. In the control group, the index of inflammation still increased one day after treatment and decreased on day 3 of treatment. After treatment, blood lactate, serum creatinine and urea nitrogen levels decreased, but the urine volume increased. After treatment, the vasoactive dose in the experimental group was significantly lower than that of the control group (P less than 0.05). CRRT is a good treatment for septic shock-related acute kidney injury, which improves biochemical indicators and protects kidney function.

The effects of nucleotide usage in key nucleotide positions +4 and -3 flanking start codon on translation levels mediated by IRES of hepatitis C virus.

Internal ribosomal entry site (IRES) functions as a cis-acting RNA element, which drives an alternative and cap-independent translation initiation pathway. Currently, there are few studies on effects of nucleotide usages at key nucleotide positions +4 and -3 flanking start codon mediated by IRES of hepatitis C virus (HCV). Herein, we focus on the effect of nucleotide usages at -3 and +4 positions mediated by HCV IRES. The nucleotide contexts flanking AUG start codon employed by HCV IRES is firstly analyzed. We found that each position in the six nucleotide positions (-4 to +6) flanking start codon of HCV has a strong tendency to select the specific nucleotide. A set of bicistronic expression vectors containing CAT gene, HCV IRES and EGFP gene were constructed, including 16 different nucleotide combinations at position -3 and +4. Each set, in which nucleotide at the -3 and +4 position has been changed into different nucleotides, included 16 types of bicistronic expression vectors. It was found that the purine nucleotide at the position -3 or +4 obviously impacts on HCV IRES-related expression, and IRES-driven translation is potentially influenced by the Kozak rule. Our results suggest that optimization of nucleotides at positions -3 and +4 is a convenient and efficient way to enhance the level of IRES-mediated translation. Keywords: Cap-independent translation; internal ribosomal entry site; hepatitis C virus; bicistronic expression vector; translation efficiency.