Programming viscoelastic properties in a complexation gel composite by utilizing entropy-driven topologically frustrated dynamical state.

Hydrogel composites in an aqueous media with viscoelastic properties and elastic modulus that can be precisely tailored are desirable to mimic many biological tissues ranging from mucus, vitreous humor, and nucleus pulposus as well as build up biosensors. Without altering the chemistry, tuning the physical interactions and structures to govern the viscoelastic properties of the hydrogels is indispensable for their applications but quite limited. Here we design a complexation gel composite and utilize the physical principle of topologically frustrated dynamical state to tune the correlated structures between the guest polycation chains and negatively charged host gels. We precisely quantify the mesh size of the host gel and guest chain size. By designing various topologically correlated structures, a viscoelastic moduli map can be built up, ranging from tough to ultrasoft, and from elastic-like with low damping properties to viscous-like with high damping properties. We also tune the swelling ratio by using entropy effect and discover an Entropy-driven Topologically Isovolumetric Point. Our findings provide essential physics to understand the relationship between entropy-driven correlated structures and their viscoelastic properties of the complexation hydrogel composites and will have diverse applications in tissue engineering and soft biomaterials.

A Fully Amorphous, Dynamic Cross-Linked Polymer Electrolyte for Lithium-Sulfur Batteries Operating at Subzero-Temperatures.

Solid-state lithium-sulfur batteries have shown prospects as safe, high-energy electrochemical storage technology for powering regional electrified transportation. Owing to limited ion mobility in crystalline polymer electrolytes, the battery is incapable of operating at subzero temperature. Addition of liquid plasticizer into the polymer electrolyte improves the Li-ion conductivity yet sacrifices the mechanical strength and interfacial stability with both electrodes. In this work, we showed that by introducing a spherical hyperbranched solid polymer plasticizer into a Li+ -conductive linear polymer matrix, an integrated dynamic cross-linked polymer network was built to maintain fully amorphous in a wide temperature range down to subzero. A quasi-solid polymer electrolyte with a solid mass content >90 % was prepared from the cross-linked polymer network, and demonstrated fast Li+ conduction at a low temperature, high mechanical strength, and stable interfacial chemistry. As a result, solid-state lithium-sulfur batteries employing the new electrolyte delivered high reversible capacity and long cycle life at 25 °C, 0 °C and -10 °C to serve energy storage at complex environmental conditions.

Shexiang Tongxin dropping pills protect against ischemic stroke-induced cerebral microvascular dysfunction via suppressing TXNIP/NLRP3 signaling pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Patients with ischemic stroke (IS) often continue to exhibit cerebral microcirculatory dysfunction even after receiving thrombolytic therapy. Enhancing the function of cerebral microvascular endothelia represents a pivotal advancement in the therapeutic strategy for ischemic microcirculatory disturbances. A traditional Chinese medicinal formulation named Shexiang Tongxin Dropping Pills (STDP), has been clinically employed to ameliorate microcirculatory abnormalities. Existing literature attests to the beneficial role of STDP on endothelial cells (ECs). Nevertheless, specific impacts and underlying mechanisms of STDP in rectifying IS-induced cerebral microvascular dysfunction warrant further exploration. AIM OF THE STUDY: This investigation seeks to delineate the effects of STDP on cerebral microvascular endothelial damage induced by ischemic stroke and to elucidate the underlying mechanism involved. MATERIALS AND METHODS: Middle cerebral artery occlusion and reperfusion (MCAO/R) technique was employed to established ischemic stroke model in mice. The therapeutic efficacy of STDP on cerebral microvascular function was assessed through laser speckle contrast imaging, behavioral assays, and histological evaluations. Biochemical markers in the brain tissue, including GSH, SOD, MDA, and ROS, were quantified using specific assay kits. In vitro study, oxygen-glucose deprivation and reperfusion (OGD/R) was performed in bEnd.3 cells. The cytoprotective potential of STDP was then evaluated by measuring cell viability, LDH activity, endothelial permeability, and oxidative stress parameters. Important targets in critical pathway were verified by immunoblotting and immunofluorescence both in mice brain slices and bEnd.3 cells. RESULTS: STDP decrease brain infarct size, repaired microvascular cerebral blood flow and attenuated neurological deficiency in MCAO/R mice. Moreover, STDP abolished MCAO/R-induced oxidative stress which was reflected by rescuing GSH content, restoration of SOD activity and T-AOC, reduction of MDA and ROS. Ex vivo, STDP increased cerebral microvascular endothelial cells viability, abolished oxidative stress and decreased their permeability after ODG/R. Mechanistically, STDP significantly suppressed endothelial ROS-TXNIP mediated the activation of NLRP3 inflammasome in vivo and in vitro. CONCLUSION: STDP improves ischemic stroke-induced cerebral microcirculatory deficits by regulating cerebral microvascular endothelial ROS/TXNIP/NLRP3 signaling pathway.

Five glycosylation-related gene signatures predict the prognostic risks of lung adenocarcinoma.

This study aimed to identify glycosylation-related genes associated with lung adenocarcinoma (LUAD) prognosis through comprehensive bioinformatic analysis. Glycosylation-related genes were identified from the Human Gene Nomenclature Committee, and LUAD prognostic genes were screened from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO)-GSE68465 datasets. Glycosylation risk score (GLRS) was calculated to predict LUAD prognostic risk. Samples were grouped into GLRS-high and GLRS-low and compared. The Tumor Immune Dysfunction and Exclusion (TIDE) score was computed to assess the antitumor immune escape possibility after immunotherapy. From 213 glycosylation-related genes, five gene signatures served as prognostic LUAD predictors using univariate and stepwise Cox regression analyses. GLRS-based models were constructed using TCGA and GSE68465 samples; their sensitivity and specificity in predicting LUAD prognosis were confirmed. GLRS was an independent LUAD prognostic factor and contributed to the nomogram to predict patient survival. High GLRS was associated with advanced tumor stage and higher mutation frequencies, estimate scores, and TIDE scores. GLRS-high and GLRS-low patients differed in immune cell infiltration and epithelial-mesenchymal transition (EMT)-related gene expression. Thus, we propose five glycosylation-related gene signatures to predict overall survival and prognostic risks of LUAD. Their regulatory roles may be related to immune invasion, immunotherapy response, mutation, and EMT.

ALCAM, Activated Leukocyte Cell Adhesion Molecule, in Clinical Gastric Cancer and Patient's Response to Chemotherapies.

BACKGROUND/AIM: Activated leukocyte cell adhesion molecule (ALCAM/CD166), a member of the immunoglobulin superfamily, has been shown to regulate cell adhesion through both homotypic and heterotypic interactions. In cancer, it might be involved in disease progression and chemotherapy drug resistance. The present study explored the clinical and prognostic significance of ALCAM in gastric cancer and its impact on patient's responses to neoadjuvant chemotherapies and cancer cells' response to chemodrugs in vitro. MATERIALS AND METHODS: Two independent cohorts were included to evaluate the link between ALCAM and the clinical outcomes and pathological factors of the patients. The gastric cancer cell lines HGC27 and AGS were used to generate ALCAM knockdown cell models. The cytotoxicity of chemotherapy drugs was examined using ALCAM knockdown cell models. RESULTS: Patients with gastric cancer who had high levels of ALCAM transcripts showed a significantly shorter overall survival in both cohorts (p=0.043 and 0.006, respectively). Patients who resisted to neoadjuvant chemotherapy had marginally higher levels of ALCAM than those responded (p=0.056). Patients with low levels of ALCAM expression and resisted to neoadjuvant chemotherapy had the worst clinical outcome with a significantly shorter overall survival (p=0.004) and disease-free survival (p=0.006), whereas such results did not appear in high ALCAM expression patients. ALCAM knockdown cells were more sensitive to Cisplatin, Oxaliplatin and 5-Fluorouracil compared with their respective control cells. CONCLUSION: ALCAM acts as a negative prognostic indicator in patients with gastric cancer and high levels of ALCAM expression result in increased chemotherapy drug resistance.

Activated Leukocyte Cell Adhesion Molecule (ALCAM), a Potential 'Seed' and 'Soil' Receptor in the Peritoneal Metastasis of Gastrointestinal Cancers.

Activated Leukocyte Cell Adhesion Molecule (ALCAM/CD166) is a cell-cell adhesion protein conferring heterotypic and homotypic interactions between cells of the same type and different types. It is aberrantly expressed in various cancer types and has been shown to be a regulator of cancer metastasis. In the present study, we investigated potential roles of ALCAM in the peritoneal transcoelomic metastasis in gastrointestinal cancers, a metastatic type commonly occurred in gastro-intestinal and gynaecological malignancies and resulting in poor clinical outcomes. Specifically, we studied whether ALCAM acts as both a 'seed' receptor in these tumour cells and a 'soil' receptor in peritoneal mesothelial cells during cancer metastasis. Gastric cancer and pancreatic cancer tissues with or without peritoneal metastasis were compared for their levels of ALCAM expression. The impact of ALCAM expression in these tumours was also correlated to the patients' clinical outcomes, namely peritoneal metastasis-free survival. In addition, cancer cells of gastric and pancreatic origins were used to create cell models with decreased or increased levels of ALCAM expression by genetic knocking down or overexpression, respectively. Human peritoneal mesothelial cells were also genetically transfected to generate cell models with different profiles of ALCAM expression. These cell models were used in the tumour-mesothelial interaction assay to assess if and how the interaction was influenced by ALCAM. Both gastric and pancreatic tumour tissues from patients who developed peritoneal metastases had higher levels of ALCAM transcript than those without. Patients who had tumours with high levels of ALCAM had a much shorter peritoneal metastasis free survival compared with those who had low ALCAM expression (p = 0.006). ALCAM knockdown of the mesothelial cell line MET5A rendered the cells with reduced interaction with both gastric cancer cells and pancreatic cancer cells. Likewise, levels of ALCAM in both human gastric and pancreatic cancer cells were also a determining factor for their adhesiveness to mesothelial cells, a process that was likely to be triggered the phosphorylation of the SRC kinase. A soluble ALCAM (sALCAM) was found to be able to inhibit the adhesiveness between cancer cells and mesothelial cells, mechanistically behaving like a SRC kinase inhibitor. ALCAM is an indicator of peritoneal metastasis in both gastric and pancreatic cancer patients. It acts as not only a potential peritoneal 'soil' receptor of tumour seeding but also a 'soil' receptor in peritoneal mesothelial cells during cancer metastasis. These findings have an important therapeutic implication for treating peritoneal transcoelomic metastases.

[Clinical effectiveness evaluation of gingival recession by using perioscope as an adjunct to non-surgical periodontal therapy].

PURPOSE: To investigate the effects of ultrasonic scaling and root planning(SRP) assisted by perioscope on gingival recession of maxillary lateral incisor. METHODS: Thirty-six outpatients with moderate to advanced chronic periodontitis from the Department of Periodontology at Dental and Ophthalmic Clinic of Putuo District from June 2020 to December 2020 were collected as research objects. Periodontal treatment was carried out according to a single-blind split-mouth self-control design randomly with(experimental group, namely perioscope group) or without(control group, namely SRP group) periodontal endoscope．The labial periodontal probing depth (PD), labial attachment loss (AL) and gingival recession(GR) in the maxillary lateral incisors were recorded at baseline, 3 and 6 months, and compared among groups by SPSS 22.0 software package. RESULTS: There was no significant difference between perioscope group and SRP group at baseline. ΔGR (the recession extent of gum within two observation time) in perioscope group was significantly smaller than that in SRP group at 3 months（P<0.05）. There was no significant difference in other periodontal indicators at 3 and 6 months between the 2 groups after treatment, but it can be found that the degree of PD reduction and AL improvement in perioscope group was more than those in SRP group, this trend was most obvious at 3 months. PD and AL were significantly different between baseline and 3 months or 6 months in the two groups. There were significant differences in ΔGR at 3 months and 6 months between the two groups. CONCLUSIONS: Compared with routine SRP, the extent of root surface debridement with perioscope-assisted SRP is thorough and less invasive, and the reduction of gingival recession of labial surface of maxillary lateral incisor at 3 months is significantly less; thus, the aesthetic effect is prominent.

VaAPRT3 Gene is Associated With Sex Determination in Vitis amurensis.

In the past decade, progress has been made in sex determination mechanism in Vitis. However, genes responsible for sexual differentiation and its mechanism in V. amurensis remain unknown. Here, we identify a sex determination candidate gene coding adenine phosphoribosyl transferase 3 (VaAPRT3) in V. amurensis. Cloning and sequencing of the VaAPRT3 gene allowed us to develop a molecular marker able to discriminate female individuals from males or hermaphrodites based on a 22-bp InDel. Gene expression and endogenous cytokinin content analysis revealed that the VaAPRT3 gene is involved in sex determination or, to be precise, in female organ differentiation, through regulating cytokinin metabolism in V. amurensis. This study enlarged the understanding of sex determination mechanism in the genus Vitis, and the sex marker could be used as a helpful tool for sexual identification in breeding programs as well as in investigation and collection of V. amurensis germplasms.

The montreal cognitive assessment and mini-mental state examination visuoexecutive subtests in acute ischemic stroke patients and their correlations with demographic and clinical factors.

Visuoexecutive impairment is common among acute ischemic stroke patients. This study aimed to examine the ability of the Montreal Cognitive Assessment (MoCA) and Mini-Mental State Examination (MMSE) visuoexecutive subtests to detect visuoexecutive abnormality in acute ischemic stroke patients and to identify the predictors for their impairments. 336 patients who completed the MMSE and MoCA were enrolled in this study. We compared the proportion of participants with incorrect MoCA visuoexecutive tasks and MMSE pentagon copying. Multivariate logistic regression analysis was used to evaluate the associations between the visuoexecutive dysfunction and demographic and clinical characteristics in the samples. Among all the participants, the MoCA detected more visuoexecutive dysfunction than the MMSE (88.69% vs. 45.83%, respectively; p < 0.001). The predictors identified by the univariate analysis included the factors of gender, age, educational level, smoking, alcohol consumption, Oxfordshire Community Stroke Project (OCSP), previous strokes, initial NIHSS score and number of old lacunar infarctions, while from the multivariate logistic regression analysis, the factors of age, educational level, NIHSS score, previous strokes and number of old lacunar infarctions served as predictive factors for the visuoexecutive impairment in acute stroke patients. In conclusion, visuoexecutive impairment is associated with the factors of the educational level, stroke severity, stroke history and number of old lacunar infarctions. Our findings may guide the clinicians to intervene the risks for the patients at an early stage after stroke and form the basis for good rehabilitation plans.

Clock-drawing test: Normative data of three quantitative scoring methods for Chinese-speaking adults in Shijiazhuang City and clinical utility in patients with acute ischemic stroke.

OBJECTIVE: The clock-drawing test (CDT) is a widely used screening tool for detecting cognitive decline. However, normative data for Chinese individuals are scarce. Our study aimed to provide standardized values for the three quantitative CDT scoring methods that were tailored for Chinese-speaking adults in Shijiazhuang City and explore the discriminant validity of the CDT scores in patients with acute ischemic stroke. METHODS: We conducted the CDT among 418 healthy individuals aged between 35 and 84 years. The CDT was administered and scored by five raters using the method derived from the Montreal Cognitive Assessment (MoCA), Rouleau's, and Babins' scoring systems. The influence of age, education, and sex on the performance in the CDT was analyzed. Furthermore, 336 patients with acute ischemic stroke were enrolled to explore the discriminant validity of CDT scores. RESULTS: In all three scoring systems, CDT scores were significantly correlated with age and years of education but not with sex. Normative data stratified for age and years of education were established. Interrater and intersystem reliability were high in our study. CDT total scores and subscores showed significant differences between stroke patients and healthy individuals. CONCLUSIONS: Our study provides CDT normative data using three quantitative scoring methods for Chinese-speaking adults in Shijiazhuang City. Age and education level were the key factors that affected the CDT scores. CDT total scores and subscores provided good discriminant validity for patients with acute ischemic stroke.

Assessing executive function following the early stage of mild Ischemic stroke with three brief screening tests.

OBJECTIVE: Executive dysfunction following stroke is well documented, but less is known about whether it occurs in mild stroke patients. The purpose of the study was to investigate executive impairment in this population and explore the correlation between executive function tests and cognitive tests of other domains. METHODS: Cross-sectional study was undertaken to compare 139 mild ischemic stroke patients (National Institute of Health Stroke Scale (NIHSS) ≤ 7) aged 40-80 with 131 normal controls matched age, gender and levels of education. All participants were administered a neuropsychological test battery including three measures of executive functioning: Clock Drawing Test (CDT), Trial Making Test-A and B (TMT-A and B), and Stroop Color Word Test (SCWT). The CDT was evaluated using three quantitative scoring rubrics, with a total score of 3,10,18, respectively and a qualitative scoring method with six types of errors. Spearman's correlations were made to analyze the correlation between executive function tests and other neuropsychological tests. RESULTS: Control group performed better than stroke group on most executive function tests at a statistical significance. Qualitative CDT showed that errors of "graphic difficulties", "conceptual deficits" and "spatial and/or planning deficits" occurred frequently in the early stage of mild stroke. Correlation data clarified that among the executive function tests, time for TMT-B correlated with global cognition most. CONCLUSION: Executive dysfunction is common following even mild strokes, and that relatively brief measures such as CDT, TMT and SCWT can be employed for it before discharge as part of rehabilitation planning.

Trends in cognitive function assessed by a battery of neuropsychological tests after mild acute ischemic stroke.

OBJECTIVE: The aim of this study is to investigate the domain-specific trends of cognitive function up to 12 months after mild acute ischemic stroke. METHODS: Enrolment of consecutive cohort of patients with mild acute ischemic stroke with recorded clinical characteristics and extensive neuropsychological assessments, including five cognitive domains. The Montreal cognitive assessment of the Beijing version (MoCA-Bj) was used to assess overall cognition. All patients completed all domain-specific examinations were categorised into three groups according to the time between the stroke onset and neuropsychological profiling, the time duration including less than one month (n = 174), one month to six months (n = 65) and over six months (n = 39). RESULTS: The final cohort consisted of 278 patients. The executive (χ2 = 6.95, P<0.05) and memory dysfunctions (χ2 = 9.6, P<0.01) showed strong improvement, especially in executive function, which prevalence was 48.85% at <1- month group and 25.64% at >6 months group. The prevalence of attention and information processing also had a declining trend, the differences, however, were not statistically significant (χ2 = 0.23 and 2.25, respectively, P>0.05). There was no significant change in language function (χ2 = 0.46, P>0.05) and the MoCA (χ2 = 0.59, P>0.05) at 3-time point groups. In 195 first-ever stroke patients, the results of memory (χ2 = 6.94 P<0.05) and executive dysfunctions (χ2 = 6.25 P<0.05) also showed significant improvement. CONCLUSION: There is varying degree of improvement tendency in executive and memory dysfunctions after mild acute ischemic stroke. Early cognitive assessments after mild acute ischemic stroke do not reflect the cognitive level of stable period.

The ratio of C2-C7 Cobb angle to T1 slope is an effective parameter for the selection of posterior surgical approach for patients with multisegmental cervical spondylotic myelopathy.

BACKGROUND: To our knowledge, laminectomy with fusion (LCF) and laminoplasty alone (LP) are both effective posterior surgical approaches for decompression of cervical spondylotic myelopathy (CSM). However, which one is suitable for patients has no standard answer. This study estimated whether the ratio of C2-C7 Cobb angle to T1 slope (CL/T1S) could be an indication of posterior surgical approach. METHODS: We retrospectively reviewed 128 patients with at least 6 months of follow-up who underwent LCF or LP. Radiological measurements, including C2-C7 Cobb angle, decompressed Cobb angle, T1 slope, cervical sagittal vertical axis, and curvature index (CI), and clinical outcomes, including Japanese Orthopedic Association score and visual analogue scale were evaluated. ROC curve analysis was used to identify discriminative power of CL/T1S ratio to predict kyphotic deformity and severe lordosis loss. The t-test and Mann-Whitney U-test were used to evaluate the difference between LCF and LP. Kruskal-Wallis H - test and ANOVA were used to evaluate the difference among different ratio CL/T1S groups. RESULTS: The cervical lordosis decreased after LCF or LP (p < 0.001, p < 0.001, respectively). Based on ROC curve analysis, CL/T1S ratio had a good discriminative power to predict kyphotic deformity and severe lordosis loss (AUC = 0.70, AUC = 0.88, respectively). According to CI value changes, cervical lordosis losses in group LP were larger than that in group LCF (p = 0.006). However, there was no significant difference in CI changes of fair-ratio CL/T1S group between LCF and LP. For patients with low CL/T1S ratio or high CL/T1S ratio, CI changes in group LP were greater than that in group LCF (p = 0.037, p = 0.042, respectively). CONCLUSIONS: CL/T1S ratio could be an indication of posterior surgical approach. Compared with LP, LCF reduces postoperative cervical lordosis losses in low-ratio and high-ratio CL/T1S groups.

[Analysis and evaluation of nutrient composition of Ophiopogonis Radix in Santai area].

Ophiopogon japonicus is one of the commonly used medicines,and it has gradually become a therapeutic food for people's daily health care. O. japonicus in Sichuan province is the famous Dao-di herbs in Sichuan province,and is mainly produced in Santai county,Sichuan province. With the unique geographical advantage,Santai county plans to declare the new food raw materials of O. japonicus based on the geographical indication products( Fu Cheng O. japonicus),so it is necessary to analyze and evaluate the nutritional components of O. japonicus in Santai county. The experimental results showed that the content of the nutrients was characterized by low fat,high polysaccharide,high potassium and high vitamin B2,which can be developed as new food raw materials.

Influence of the Ratio of C2-C7 Cobb Angle to T1 Slope on Cervical Alignment After Laminoplasty.

PURPOSE: To assess the relationship between the ratio of C2-C7 Cobb angle to T1 slope (CL/T1S) and cervical alignment changes after laminoplasty. METHODS: 78 consecutive patients with cervical myelopathy who underwent laminoplasty were enrolled. All patients with preoperative and follow-up cervical spine lateral x-ray images available for review were recruited in this study. Imaging data included C2-C7 Cobb angle, T1 slope, and cervical sagittal vertical axis (cSVA). All patients were classified into low-ratio group (bottom 25% of CL/T1S), fair-ratio group (middle 50% of CL/T1S), and high ratio group (top 25% of CL/T1S) according to CL/T1S ratio. The recovery rate was calculated based on the Japanese Orthopedic Association score. RESULTS: The preoperative C2-C7 Cobb angle had significant correlations with the T1 slope (r = 0.528). Kyphotic alignment changes in the group with a high ratio of CL/T1S was greater than that of the other 2 groups (P < 0.001). The incidence of postoperative kyphosis in the group with a low ratio of CL/T1S was higher than that of the other 2 groups (P < 0.001). There was no postoperative kyphosis in the fair-ratio group. The surgical outcome in the low-ratio CL/T1S group and the high-ratio CL/T1S group was poorer than that in the fair-ratio CL/T1S group (P = 0.005). CONCLUSIONS: The cervical alignment was kept well in the mid-range CL/T1S ratio group after laminoplasty. Patients with a high CL/T1S ratio were more likely to present with kyphotic alignment changes. Patients with a low CL/T1S ratio were more likely to have postoperative kyphosis.

Clinical and magnetic resonance imaging predictors of the surgical outcomes of patients with cervical spondylotic myelopathy.

OBJECTIVE: To determine whether clinical characteristics and signal and morphologic changes on magnetic resonance (MR) images of the spinal cord (SC) are associated with surgical outcomes for cervical spondylotic myelopathy (CSM). PATIENTS AND METHODS: A total of 113 consecutive patients with cervical myelopathy underwent cervical decompression surgery in our hospital from January 2015 to January 2018. All patients with preoperative MR images available for review were recruited for this study. Research data included patient sex, age, duration of symptoms, surgical approach, compression level, preoperative mJOA (modified Japanese Orthopaedic Association) score, postoperative mJOA recovery rate, and complications. Imaging data included signal changes on T2-weighted MRI images (grade and extension on sagittal images, four types of signal changes on axial images according to the Ax-CCM system), SC compression, transverse area of the SC, and compression ratio. The t-test, Mann-Whitney U-test, Kruskal-Wallis H - test, analysis of variance, and regression analysis were used to evaluate the effects of individual predictors on surgical outcomes. RESULTS: The study cohort included 85 males and 27 females with a mean age of 60.92 ± 8.93 years. The mean mJOA score improved from 10.24 ± 1.69 preoperatively to 15.11 ± 2.05 at the final follow-up (p < 0.001). Patients in the poor outcome group were more likely to present with a longer duration of symptoms (p < 0.001) and smaller transverse area of the SC (p < 0.001). Bright T2-weighted high signal changes (T2HSCs), multisegmental high signal changes on sagittal MR images, and fuzzy focal T2HSCs on axial MR images were associated with a poor outcome (p < 0.001, p = 0.005, p < 0.001, respectively). The maximum SC compression and compression ratio were not reliable predictors of surgical outcomes (p = 0.375, p = 0.055, respectively). The result of multivariate stepwise logistic regression showed that a longer duration of symptoms, multisegmental T2HSCs on sagittal MR images and fuzzy focal T2HSCs on axial MR images were significant risk factors of poor outcomes (p < 0.001, p = 0.049, p = 0.016, respectively). CONCLUSION: A longer duration of symptom, multisegmental T2HSCs on sagittal MR images, and fuzzy focal T2HSCs on axial MR images were highly predictive of a poor surgical outcome for CSM. Smaller transverse area of the SC and bright T2HSCs were also associated with the prognosis of CSM.

[Micro-CT study of root canal morphology of mandibular first premolars with multi-canals].

PURPOSE: To observe root canal system of multi-canal mandibular first premolars and to analyze the morphology of them by micro-CT and three-dimensional reconstruction. METHODS: Thirty-two extracted mandibular first premolars with multiple root canals were selected and scanned by micro-CT. The number and morphology of canals, location of bifurcation and 5 subtle structures (lateral canals, apical ramification, isthmus, loops, apical delta) were analyzed. RESULTS: In the 32 specimens, 96.88% contained radicular depressions or grooves. The canal configuration was type II, Ⅲ, Ⅴ: 3.12%, type Ⅳ: 31.25%, type Ⅷ: 9.38%, type Ⅸ: 15.62% and C-shaped canal: 34.38% by Vertucci's classification. 62.50% of the canals bifurcated at the middle third of roots, comparing with 37.50% at apical third. The incidence of the lateral canals, apical ramification, isthmus, loops and apical delta was 56.25%, 43.75%, 40.62%, 21.88% and 6.25%, respectively, which were all found at the middle and apical third of roots except that lateral canals were found at all depth of canals. CONCLUSIONS: Most mandibular first premolars with multiple canals contain radicular depression or grooves, of which root canal system are complicated. Micro-CT can provide clear and accurate three-dimensional information of canal morphology.

Production of 3-hydroxypropionic acid in engineered Methylobacterium extorquens AM1 and its reassimilation through a reductive route.

BACKGROUND: 3-Hydroxypropionic acid (3-HP) is an important platform chemical, serving as a precursor for a wide range of industrial applications such as the production of acrylic acid and 1,3-propanediol. Although Escherichia coli or Saccharomyces cerevisiae are the primary industrial microbes for the production of 3-HP, alternative engineered hosts have the potential to generate 3-HP from other carbon feedstocks. Methylobacterium extorquens AM1, a facultative methylotrophic α-proteobacterium, is a model system for assessing the possibility of generating 3-HP from one-carbon feedstock methanol. RESULTS: Here we constructed a malonyl-CoA pathway by heterologously overexpressing the mcr gene to convert methanol into 3-HP in M. extorquens AM1. The engineered strains demonstrated 3-HP production with initial titer of 6.8 mg/l in shake flask cultivation, which was further improved to 69.8 mg/l by increasing the strength of promoter and mcr gene copy number. In vivo metabolic analysis showed a significant decrease of the acetyl-CoA pool size in the strain with the highest 3-HP titer, suggesting the supply of acetyl-CoA is a potential bottleneck for further improvement. Notably, 3-HP was rapidly degraded after the transition from exponential phase to stationary phase. Metabolomics analysis showed the accumulation of intracellular 3-hydroxypropionyl-CoA at stationary phase with the addition of 3-HP into the cultured medium, indicating 3-HP was first converted to its CoA derivatives. In vitro enzymatic assay and ß-alanine pathway dependent 13C-labeling further demonstrated that a reductive route sequentially converted 3-HP-CoA to acrylyl-CoA and propionyl-CoA, with the latter being reassimilated into the ethylmalonyl-CoA pathway. The deletion of the gene META1_4251 encoding a putative acrylyl-CoA reductase led to reduced degradation rate of 3-HP in late stationary phase. CONCLUSIONS: We demonstrated the feasibility of constructing the malonyl-CoA pathway in M. extorquens AM1 to generate 3-HP. Furthermore, we showed that a reductive route coupled with the ethylmalonyl-CoA pathway was the major channel responsible for degradation of the 3-HP during the growth transition. Engineered M. extorquens AM1 represents a good platform for 3-HP production from methanol.

Synergistic anticancer effect of exogenous wild-type p53 gene combined with 5-FU in human colon cancer resistant to 5-FU in vivo.

AIM: To investigate the anticancer effect of a recombinant adenovirus-mediated p53 (rAd-p53) combined with 5-fluorouracil (5-FU) in human colon cancer resistant to 5-FU in vivo and the mechanism of rAd-p53 in reversal of 5-FU resistance. METHODS: Nude mice bearing human colon cancer SW480/5-FU (5-FU resistant) were randomly assigned to four groups (n = 25 each): control group, 5-FU group, rAd-p53 group, and rAd-p53 + 5-FU group. At 24 h, 48 h, 72 h, 120 h and 168 h after treatment, 5 mice were randomly selected from each group and sacrificed using an overdose of anesthetics. The tumors were removed and the protein expressions of p53, protein kinase C (PKC), permeability-glycoprotein (P-gp) and multidrug resistance-associated protein 1 (MRP1) (Western blot) and apoptosis (TUNEL) were determined. RESULTS: The area ratios of tumor cell apoptosis were larger in the rAd/p53 + 5-FU group than that in the control, 5-FU and rAd/p53 groups (P < 0.05), and were larger in the rAd/p53 group than that of the control group (P < 0.05) and the 5-FU group at more than 48 h (P < 0.05). The p53 expression was higher in the rAd/p53 and the rAd/p53 + 5-FU groups than that of the control and 5-FU groups (P < 0.05), and were higher in the rAd/p53 + 5-FU group than that of the rAd/p53 group (P < 0.05). Overexpression of PKC, P-gp and MRP1 was observed in the 5-FU and control groups. In the rAd/p53 + 5-FU group, the expression of P-gp and MRP1 was lower that of the control and 5-FU groups (P < 0.05), and the expression of PKC was lower than that of the control, 5-FU and rAd/p53 groups at more than 48 h (P < 0.05). In the rAd/p53 group, the expression of P-gp and MRP1 was lower that of the control and 5-FU groups at more than 48 h (P < 0.05), and the expression of PKC was lower than that of the control and 5-FU groups at more than 120 h (P < 0.05). CONCLUSION: 5-FU combined with rAd-p53 has a synergistic anticancer effect in SW480/5-FU (5-FU resistance), which contributes to reversal of 5-FU resistance.