Investigation of the Properties of High-Viscosity Modified Asphalt Binder under Hygrothermal Environments.

High-viscosity modified asphalt binder (HVMA) is used widely as a polymer-modified binder in porous asphalt pavement because it can improve the cohesiveness of the asphalt mixture. However, because of the high voidage in the mixture, HVMA is vulnerable to aging induced by temperature, oxygen, water, sunlight, and other climatic conditions, which degrades the performance of pavement. The properties of asphalt binder are affected adversely by the effects of hygrothermal environments in megathermal and rainy areas. Therefore, it is essential to study the aging characteristics of HVMA under the influence of hygrothermal environments to promote its application as a high-viscosity modifier. A hygrothermal cycle aging test (HCAT) was designed to simulate the aging of HVMA when rainwater was kept inside of the pavement after rainfall in megathermal areas. One kind of base bitumen and three kinds of HVMA (referred to as SBS, A, and B, respectively) were selected in this study. Short-term aging tests, hygrothermal cycling aging tests, and long-term aging tests were performed on the base bitumen and three kinds of modified asphalt binder. Fourier-transform infrared spectroscopy (FTIR), thermo-gravimetric analysis (TGA), and dynamic shear rheological (DSR) tests were used to evaluate the properties of the binders on the micro and macro scales. By comparing the index variations of the four binders before and after aging, the effects of the hygrothermal environment on the properties of HVMA were studied. It was found that the effects of the hygrothermal environment expedited the decomposition of the polymer and the formation of carbonyl groups compared with the TFOT and PAV test, which TGA confirmed further. Moreover, the thermal stability of the samples was improved after HCAT. In addition, the master curves of the complex modulus showed that hygrothermal cycles made the high-temperature rutting resistance of asphalt binder increase significantly. All of the results above verified that the effect of hygrothermal cycling could accelerate the aging of HVMA and shorten its service life.

An innovative cell-based transplantation therapy for an immature permanent tooth in an adult: a case report.

BACKGROUND: Immature teeth with necrotic pulps present multiple challenges to clinicians. In such cases, regenerative endodontic procedures (REPs) may be a favorable strategy. Cells, biomaterial scaffolds, and signaling molecules are three key elements of REPs. Autologous human dental pulp cells (hDPCs) play an important role in pulp regeneration. In addition, autologous platelet concentrates (APCs) have recently been demonstrated as effective biomaterial scaffolds in regenerative dentistry, whereas the latest generation of APCs-concentrated growth factor (CGF), especially liquid phase CGF (LPCGF)-has rarely been reported in REPs. CASE PRESENTATION: A 31-year-old woman presented to our clinic with the chief complaint of occlusion discomfort in the left mandibular posterior region for the past 5 years. Tooth #35 showed no pulp vitality and had a periodontal lesion, and radiographic examination revealed that the tooth exhibited extensive periapical radiolucency with an immature apex and thin dentin walls. REP was implemented via transplantation of autologous hDPCs with the aid of LPCGF. The periodontal lesion was managed with simultaneous periodontal surgery. After the treatment, the tooth was free of any clinical symptoms and showed positive results in thermal and electric pulp tests at 6- and 12-month follow-ups. At 12-month follow-up, radiographic evidence and three-dimensional models, which were reconstructed using Mimics software based on cone-beam computed tomography, synergistically confirmed bone augmentation and continued root development, indicating complete disappearance of the periapical radiolucency, slight lengthening of the root, evident thickening of the canal walls, and closure of the apex. CONCLUSION: hDPCs combined with LPCGF represents an innovative and effective strategy for cell-based regenerative endodontics.

Construction and Evaluation of the Immunogenicity and Protective Efficacy of Recombinant Replication-Deficient Human Adenovirus-5 Expressing Genotype VII Newcastle Disease Virus F Protein and Infectious Bursal Disease Virus VP2 Protein.

Newcastle disease (ND) and infectious bursal disease (IBD) are two key infectious diseases that significantly threaten the health of the poultry industry. Although existing vaccinations can effectively prevent and treat these two diseases through multiple immunizations, frequent immunization stresses significantly impact chicken growth. In this study, three recombinant adenoviruses, rAd5-F expressing the NDV (genotype VII) F protein, rAd5-VP2 expressing the IBDV VP2 protein, and rAd5-VP2-F2A-F co-expressing F and VP2 proteins, were constructed using the AdEasy system. The F and VP2 genes of the recombinant adenoviruses could be transcribed and expressed normally in HEK293A cells as verified by RT-PCR and Western blot. The three recombinant viruses were shown to have similar growth kinetics as rAd5-EGFP. Compared with the PBS and rAd5-EGFP groups, SPF chickens immunized with recombinant adenoviruses produced higher antibody levels, more significant lymphocyte proliferation, and significantly higher CD4+/CD3+ and CD8+/CD3+ cells in peripheral blood. The survival rate of SPF chickens immunized with rAd5-F and rAd5-VP2-F2A-F after the challenge with DHN3 was 100%, and 86% of SPF chickens showed no viral shedding at 7 dpc. The survival rate of SPF chickens immunized with rAd5-VP2 and rAd5-VP2-F2A-F after the challenge with BC6/85 was 86%. rAd5-VP2 and rAd5-VP2-F2A-F significantly inhibited bursal atrophy and pathological changes compared to the rAd5-EGFP and PBS groups. This study provides evidence that these recombinant adenoviruses have the potential to be developed into safe and effective vaccine candidates for the prevention and control of ND and IBD.

CRISPR/Cas9 Gene Editing of NtAITRs, a Family of Transcription Repressor Genes, Leads to Enhanced Drought Tolerance in Tobacco.

Tobacco is a cash crop throughout the world, and its growth and development are affected by abiotic stresses including drought stress; therefore, drought-tolerant breeding may help to improve tobacco yield and quality under drought stress conditions. Considering that the plant hormone ABA (abscisic acid) is able to regulate plant responses to abiotic stresses via activating ABA response genes, the characterization of ABA response genes may enable the identification of genes that can be used for molecular breeding to improve drought tolerance in tobacco. We report here the identification of NtAITRs (Nicotiana tabacum ABA-induced transcription repressors) as a family of novel regulators of drought tolerance in tobacco. Bioinformatics analysis shows that there are a total of eight NtAITR genes in tobacco, and all the NtAITRs have a partially conserved LxLxL motif at their C-terminus. RT-PCR results show that the expression levels of at least some NtAITRs were increased in response to ABA and drought treatments, and NtAITRs, when recruited to the Gal4 promoter via a fused GD (Gal4 DNA-binding domain), were able to repress transcription activator LD-VP activated expression of the LexA-Gal4-GUS reporter gene. Roles of NtAITRs in regulating drought tolerance in tobacco were analyzed by generating CRISPR/Cas9 gene-edited mutants. A total of three Cas9-free ntaitr12356 quintuple mutants were obtained, and drought treatment assays show that drought tolerance was increased in the ntaitr12356 quintuple mutants. On the other hand, results of seed germination and seedling greening assays show that ABA sensitivity was increased in the ntaitr12356 quintuple mutants, and the expression levels of some ABA signaling key regulator genes were altered in the ntaitr12356-c3 mutant. Taken together, our results suggest that NtAITRs are ABA-responsive genes, and that NtAITRs function as transcription repressors and negatively regulate drought tolerance in tobacco, possibly by affecting plant ABA response via affecting the expression of ABA signaling key regulator genes.

Low Prognostic Nutritional Index Contributes to High Adverse Events in Preeclampsia.

Background: Preeclampsia (PE) is a common obstetric complication that has caused significant harm to pregnant mothers. The clinical significance of poor nutritional status in PE patients is unclear. The aim of our study was to evaluate the nutritional status as measured by the prognostic nutritional index (PNI) score at admission, and its ability to predict in-hospitalization adverse events in patients with PE. Methods: We enrolled patients diagnosed with PE in the Third Affiliated Hospital of Guangzhou Medical University from January 2019 to December 2021. Patients were divided into low and high nutritional status group according to the cut-off value of PNI score at admission using the receiver operating characteristic (ROC) curve. PNI score were used to explore the relationship between PNI score and in-hospitalization adverse events presented with hazard ratio (HR) and 95% confidence intervals (CI). Results: A total of 733 patients were included in the study. The proportion of adverse events and admission to intensive care unit (ICU) was higher in the low nutritional status group than in the high nutritional status group (P < 0.05). ROC curve analysis revealed an area under curve (AUC) of 0.628 for PNI score and the cut-off value of PNI was 37. The free-event rates determined by KM analysis were significantly lower in the low nutritional status compared to the high nutritional status (P < 0.05). Adjusted multivariate analysis showed that PNI score was independently associated with favorable outcomes (HR: 2.66; 95% CI: 1.724-4.050, P < 0.001). Conclusion: High PNI score at admission was associated with reduced in-hospitalization risk of adverse events in patients with PE. Additional enhancing nutritional status during hospitalization may help to prevent unfavorable prognosis in clinical practices.

Mutation of GmAITR Genes by CRISPR/Cas9 Genome Editing Results in Enhanced Salinity Stress Tolerance in Soybean.

Breeding of stress-tolerant plants is able to improve crop yield under stress conditions, whereas CRISPR/Cas9 genome editing has been shown to be an efficient way for molecular breeding to improve agronomic traits including stress tolerance in crops. However, genes can be targeted for genome editing to enhance crop abiotic stress tolerance remained largely unidentified. We have previously identified abscisic acid (ABA)-induced transcription repressors (AITRs) as a novel family of transcription factors that are involved in the regulation of ABA signaling, and we found that knockout of the entire family of AITR genes in Arabidopsis enhanced drought and salinity tolerance without fitness costs. Considering that AITRs are conserved in angiosperms, AITRs in crops may be targeted for genome editing to improve abiotic stress tolerance. We report here that mutation of GmAITR genes by CRISPR/Cas9 genome editing leads to enhanced salinity tolerance in soybean. By using quantitative RT-PCR analysis, we found that the expression levels of GmAITRs were increased in response to ABA and salt treatments. Transfection assays in soybean protoplasts show that GmAITRs are nucleus proteins, and have transcriptional repression activities. By using CRISPR/Cas9 to target the six GmAITRs simultaneously, we successfully generated Cas9-free gmaitr36 double and gmaitr23456 quintuple mutants. We found that ABA sensitivity in these mutants was increased. Consistent with this, ABA responses of some ABA signaling key regulator genes in the gmaitr mutants were altered. In both seed germination and seedling growth assays, the gmaitr mutants showed enhanced salt tolerance. Most importantly, enhanced salinity tolerance in the mutant plants was also observed in the field experiments. These results suggest that mutation of GmAITR genes by CRISPR/Cas9 is an efficient way to improve salinity tolerance in soybean.

N-terminal truncated carboxypeptidase E represses E-cadherin expression in lung cancer by stabilizing the Snail-HDAC complex.

The N-terminal truncated carboxypeptidase E (CPEΔN) protein, an alternative splicing product of the carboxypeptidase E gene, has recently been recognized as an independent predictor for the recurrence and metastasis of lung adenocarcinoma. In this study, we showed that CPEΔN may accelerate lung cancer invasion via an E-cadherin-dependent mechanism. In vitro experiments and in vivo bioluminescence imaging assay revealed CPEΔN promoted the mobility and invasion of human lung cancer cells by suppressing endogenous expression of E-cadherin, a critical regulator for epithelial tissue homeostasis. Further mechanistic analyses revealed that CPEΔN directly interacted with and stabilized the Snail/HDAC1/HDAC3 complex within the promoter region of the E-cadherin-encoding CDH1 gene. CPEΔN overexpression led to a reduction of histone H3K9 acetylation and an increase of H3K9 and H3K27 trimethylation in the CHD1 gene promoter and ultimately inhibited E-cadherin transcription. In addition, correlations among CPEΔN, E-cadherin expression and tumor progression in 195 cases of lung adenocarcinoma patients were analyzed. Higher nuclear expression of CPEΔN was detected in patients with advanced stage of lung adenocarcinoma. Nuclear expression of CPEΔN was negatively correlated with the cell membrane expression of E-cadherin. Collectively, our findings illustrated that CPEΔN was involved in the transcriptional regulation of the epithelial-mesenchymal transition-related gene CDH1 and provide novel insights into CPEΔN-associated lung cancer metastasis.

[Early age-related macular degeneration treated with emayaoling acupuncture technique: a randomized controlled trial].

OBJECTIVE: To compare the differences in the clinical therapeutic effects on early age-related macular degeneration (AMD) in the treatment between emayaoling acupuncture technique and regular acupuncture. METHODS: A total of 110 patients of AMD were randomized into an observation group (55 cases, 73 eyes) and a control group (55 cases, 76 eyes). In the observation group, acupuncture was applied at Cuanzhu (BL 2) and Yiming (EX-HN 14). The needles were retained for 30 min. Additionally, Ganshu (BL 18), Pishu (BL 20) and Shenshu (BL 23) were stimulated with quick needling technique. Emayaoling needling technique was applied. The needle was rotated with the thumb and index finger of the puncture hand, forward and backward. The strong and large amplitude were given when the thumb moved forward, and while, the week and small amplitude required when moving backward. The gentle movement was required in the entire procedure. In the control group, the point selection was same as the observation group. The points were stimulated with regular technique. The needle retaining time was same as the observation group. The treatment was given once every two days in the two groups and for 3 months continuously. The visit was followed up for 3 months. Before and after treatment, as well as at follow-up, the eyesight, the effect on macular area and the macular retinal structure [the macular nerve fiber layer (MNFL), retinal neurepithelium layer (RNL), pigment epithelium and choroid capillary composite lay (PECCL)] were evaluated separately. RESULTS: 1. After treatment and at follow-up, the eyesight was improved as compared with that before treatment in the two groups, indicating the significant difference (both P<0.05). The differences in comparison before and after treatment, and in comparison of the follow-up with that before treatment, were improved in a larger amplitude in the observation group as compared with the control group, indicating the significant difference (both P<0.05). 2. Regarding the therapeutic effects on macular area, after treatment and at follow-up, the total effective rates were 83.6% (61/73) and 86.3% (63/73) in the observation group and were 63.2% (48/76) and 67.1% (51/76) in the control group separately. The results in the observation group were higher than those in the control group, indicating the significant difference between the two groups (both P<0.05). 3. Regarding the macular retinal structure, after treatment and at follow-up, the levels of MNFL, RNL and PECCL were all reduced as compared with those before treatment, indicating the significant difference (all P<0.05). After treatment and at follow-up, the levels of MNFL, RNL and PECCL in the observation group were all lower than those in the control group, indicating the significant difference (all P<0.05). CONCLUSION: The emayaoling acupuncture technique achieves better clinical therapeutic effects on treatment of early AMD as compared with the regular acupuncture.

Digital Anatomical Measurement for Anterolateral Fixation of Middle and Lower Thoracic Vertebrae.

BACKGROUND The key to its successful application is to determine the best entry point for the vertebral screw(s). This study aimed to provide a reference for clinical anterolateral fixation through digital measurement of computed tomography (CT) data to identify relevant anatomical positions in the middle and lower thoracic vertebrae (T4-T12) of 30 adults. MATERIAL AND METHODS We performed digital measurement of anatomical positions in the middle and lower thoracic vertebrae (T4-T12) of 30 adults. ABBREVIATIONS: Left height of vertebral body, LHV; Right height of vertebral body, RHV; Anterior height of vertebral body, AHV; Middle height of vertebral body, MHV; Posterior height of vertebral body, PHV; Superior sagittal diameter of vertebral body, SSDV; Superior transverse diameter of vertebral body, STDV; inferior sagittal diameter of vertebral body, ISDV; Inferior transverse diameter of vertebral body, ITDV; (1) Left (right) height of vertebral body, [L(R)HV]; Anterior (middle, posterior) height of vertebral body [A(M,P)HV]; Superior (inferior) sagittal diameter of vertebral body, [S(I)SDV]; Superior (inferior) transverse diameter of vertebral body, [S(I)TDV]. RESULTS The transverse diameters of vertebral bodies were always larger than the sagittal diameter for 3~4 mm. The distance between 2 vertebrae (interval of 1 vertebra) range were (52-56) mm for T4-T7 and (44-48) mm for T8-T12, and the surgeons could collate these data to choose a suitable stick length. CONCLUSIONS Bone graft should prune into laterigrade cuboid, it can recover A-P and bilateral physiological functions load, and the height of the vertebral body increased from T4 to T12.

Selection of bisphenol A - single-chain antibodies from a non-immunized mouse library by ribosome display.

Developing reagents with high affinity and specificity are critical to detect the environmental hormones or toxicants. Ribosome display technology has been widely used in functional protein or peptide screening and in directed evolution of protein molecules in vitro. In this study, single-chain variable fragments (scFvs) against bisphenol A (BPA) were selected from a library constructed from splenocytes of non-immunized mice. After five rounds of selection, the selected scFvs bound to BPA with high affinity. Indirect competitive enzyme-linked immunosorbent assay (ELISA) was introduced to screen the antibody affinity and specificity to BPA. The equilibrium dissociation constants (KDS) of one clone was 1.76µM as determined by surface plasmon resonance (SPR). This study indicated that ribosome display can isolate binders to small molecules from a non-immunized naive library without any in vivo steps and can generate recombinant antibodies efficiently and rapidly. In addition, this study provides a methodological framework for detection of small molecules using recombinant antibodies.

Executive dysfunction in patients with cerebral hypoperfusion after cerebral angiostenosis/occlusion.

Impairment of executive functions (EFs) was investigated in patients with cerebral hypoperfusion after cerebral angiostenosis/occlusion. Several EFs were measured in patients with cerebral angiostenosis/occlusion and healthy subjects. The vascular conditions, regional cerebral blood flow (rCBF), regional cerebral blood volume (rCBV), mean transit time (MTT), time to peak (TTP), and delay time were assessed. The scores of the vascular stenosis/occlusion group were significantly lower than those of the control group. rCBV and rCBF were negatively correlated with the error response times in the Stroop test, and the persistent error responses in the Wisconsin Card Sorting Test (WCST) were positively correlated with the Montreal Cognitive Assessment (MoCA) scores. TTP was positively correlated with the reaction and error reaction times, and the persistent error response in WCST was negatively correlated with the times of sorting in WCST and MoCA scores. MTT was positively correlated with the persistent error response in WCST. In the Stroop test, delay time was positively correlated with response time, and negatively correlated with error response times, and the persistent error response in WCST and MoCA scores. Patients with cerebral hypoperfusion after cerebral angiostenosis/occlusion had executive dysfunctions in working memory, sustained attention, response inhibition, cognitive flexibility, thought organization, planning, and implementation. Moreover, their executive dysfunctions were related with the decline in rCBF and rCBV. The prolonged TTP, MTT, and delay time suggested a slow blood flow and the poor compensation of collateral circulation, resulting in impairment of the EFs.

Protocol for the use of light upon extension real-time PCR for the determination of viral load in HBV infection.

Real-time PCR has engendered wide acceptance for quantitation of hepatitis B virus (HBV) DNA in the blood due to its improved rapidity, sensitivity, reproducibility, and reduced contamination. Here we describe a cost-effective and highly sensitive HBV real-time quantitative assay based on the light upon extension real-time PCR platform and a simple and reliable HBV DNA preparation method using silica-coated magnetic beads.

A novel real-time PCR assay for determination of viral loads in person infected with hepatitis B virus.

A novel LUX (Light Upon eXtension) primer-based real-time PCR assay was developed and evaluated in this study, which was designed to provide a cost-effective, specific and highly sensitive method for viral load determination of hepatitis B virus (HBV). The assay employed an effective and rapid nucleic acid extraction system based on magnetic beads. To evaluate its efficacy, this new viral DNA preparation method was compared with QIAamp Blood Mini Kit and the results showed a good correlation (r=0.971; P<0.001). The performance of the LUX real-time assay was validated by testing serial dilutions of HBV plasmid DNA (5 to 5 x 10(8)copies/reaction) and a good linear relationship was obtained between the Ct values and the log(10) concentration of the HBV DNA. The assay possessed high sensitivity and the detection limit of this system was as few as 25 copies/ml of serum. A total of 91 positive serum samples were detected to evaluate further the assay and the high specificity was confirmed by melting curve analysis. This assay provides an ideal tool for monitoring the treatment efficacy and studying the relationship between HBV viral load and the stage of disease.