Radiographic adjacent segment degeneration and risk factors for osteoporotic vertebral compression fractures treated with percutaneous kyphoplasty.

PURPOSE: In this study, we aimed to clarify whether osteoporotic vertebral compression fracture (OVCF) following percutaneous kyphoplasty (PKP) was associated with a high risk for radiographic adjacent segment degeneration (ASD) and to identify the risk factors for radiographic ASD in these individuals. METHODS: We retrospectively reviewed consecutive patients with OVCFs who underwent PKP at our institution between November 2015 and January 2021. The incidence of radiographic ASD was calculated and specific subgroups of ASD were identified. Univariate and multivariate analyses of demographic, clinical baseline, and radiologic data were performed to identify risk factors associated with radiographic ASD. RESULTS: With a mean follow-up time of 27.3 months, a total of 95 eligible patients were enrolled. The incidence of radiographic ASD distinguished from natural degeneration was 52.6%. Patients with OVCFs who underwent PKP had a high risk of developing radiographic ASD, particularly disc degeneration. Intradiscal cement leakage (odds ratio [OR], 5.706; 95% confidence interval [CI], 2.039-15.970; P = 0.001) and preoperative disc height (OR, 0.681; 95% CI, 0.518-0.895; P = 0.006) were identified as independent risk factors. CONCLUSION: Patients with OVCFs who underwent PKP were more likely to develop radiographic ASD, and their progression was distinguished from natural degeneration. Disc degeneration was the most common type of degeneration. Intradiscal cement leakage and preoperative disc height were identified as independent risk factors for developing radiographic ASD in these patients. Further validation through prospective multicenter studies is required.

A multiplex one-step fluorescence quantitative differential diagnosis method for severe hand, foot and mouth disease caused by coxsackievirus A16.

Hand foot and mouth disease (HFMD) is a common childhood infectious disease which is caused by human enterovirus. The objective of this study was to develop a rapid, sensitive, and accurate method for detecting severe HFMD caused by coxsackievirus A16 (CV-A16). A closed-tube sensitive multiplex one-step reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) was applied to detect CV-A16 in the early stage of severe HFMD. This assay targeted the CV-A16 structure protein VP1 to distinguish CV-A16 from other coxsackieviruses The 5'UTR region of enteric viruses was used for detecting the enterovirus and ribonuclease P (RNaseP) was adopted as the internal reference gene. The multiplex MGB probe assay system was used to detect PCR amplicons with different fluorescence reporters in the same system. The limit of detection (LOD) of the RT-qPCR assay for the CV-A16 VP1 gene was 125.893 copies/µl, for the 5' UTR was 50.1187 copies/µl and for the RNaseP gene was 158.49 copies/µl. Furthermore, specificity analysis showed that the multiplex RT-PCR had no cross-reactivity with the influenza virus, herpangina virus and SARS-COV-2. In correlation analysis, the sensitivity of the multiplex RT-qPCR assay for CV-A16 detection was 100 % (288/288) and the specificity of the multiplex RT-qPCR assay was 99.94 % (3395/3397). The overall agreement between the multiplex RT-qPCR and the results of clinical diagnosis was 99.95 % (3683/3685) and kappa value was 0.996 (p<0.001). The entire procedure, from specimen processing to result reporting, could be completed within 1.5 hours. The one-step multiplex RT-qPCR assay for detecting CV-A16 developed in this study is a good laboratory diagnostic tool for rapid and reliable distinguished detection of CV-A16, especially for severe HFMD patients at an early stage in the disease with low virus load of CV-A16.

[Effect of polishing method and time on the fitness of CAD/CAM zirconia ceramic crown].

PURPOSE: To compare the effect of different polishing methods and time treatment on the fitness of CAD/CAM zirconia ceramic crowns. METHODS: Sixteen intact maxillary premolars were randomly divided into two groups, group A was treated with silicon carbide burs, while group B was treated with tungsten steel burs. At different polishing time points of the same tooth, digital impressions of each group were obtained, which were used to manufacture CAD/CAM zirconium ceramic crowns. After trial fitting, the gap impressions were obtained by using silicone rubber replication method, and the marginal and internal discrepancies were assessed. The data were statistically analyzed with SPSS 21.0 software package. RESULTS: The difference between the gap values of the marginal and internal markers of group A and group B was not statistically significant(P＞0.05). Compared with the no-polishing process, the differences of the marginal gap (39.67±8.35) µm and internal gap (45.18±7.16) µm of group A polished for 4 min, and the marginal gap (51.25±14.73) µm, and internal gap (48.56±6.45) µm of group B polished for 3 min, as well as the marginal gap (48.87±8.90) µm, and internal gap (45.99±7.12) µm of group B polished for 4 min, were all significant(P＜0.05). CONCLUSIONS: CAD/CAM zirconia ceramic crowns treated with silicon carbide bur for polishing 4 min and tungsten steel for 3 min has the best fitness.

Genotype-phenotype pattern analysis of pathogenic PAX9 variants in Chinese Han families with non-syndromic oligodontia.

Background: Non-syndromic oligodontia is characterized by the absence of six or more permanent teeth, excluding third molars, and can have aesthetic, masticatory, and psychological consequences. Previous studies have shown that PAX9 is associated with autosomal dominant forms of oligodontia but the precise molecular mechanisms are still unknown. Methods: Whole-exome and Sanger sequencing were performed on a cohort of approximately 28 probands with NSO, for mutation analysis. Bioinformatic analysis was performed on the potential variants. Immunofluorescence assay, western blotting, and qPCR were used to explore the preliminary functional impact of the variant PAX9 proteins. We reviewed PAX9-related NSO articles in PubMed to analyze the genotype-phenotype correlations. Results: We identified three novel PAX9 variants in Chinese Han families: c.152G>T (p.Gly51Val), c.239delC (p.Thr82Profs*3), and c.409C>T (q.Gln137Ter). In addition, two previously reported missense variants were identified: c.140G>C (p.Arg47Pro) and c.146C>T (p.Ser49Leu) (reference sequence NM_006194.4). Structural modeling revealed that all missense variants were located in the highly conserved paired domain. The other variants led to premature termination of the protein, causing structural impairment of the PAX9 protein. Immunofluorescence assay showed abnormal subcellular localizations of the missense variants (R47P, S49L, and G51V). In human dental pulp stem cells, western blotting and qPCR showed decreased expression of PAX9 variants (c.140G>C, p.R47P, and c.152G>T, p.G51V) compared with the wild-type group at both the transcription and translation levels. A review of published papers identified 64 PAX9 variants related to NSO and found that the most dominant feature was the high incidence of missing upper second molars, first molars, second premolars, and lower second molars. Conclusion: Three novel PAX9 variants were identified in Chinese Han families with NSO. These results extend the variant spectrum of PAX9 and provide a foundation for genetic diagnosis and counseling.

Functionalized pyrite nanozyme probe and imprinted polymer modified with hydrophilic layer for rapid colorimetric analysis of glycoprotein in serum.

The biological molecules used in the sandwich detection method have problems such as complex extraction processes, high costs, and uneven quality. Therefore we integrated glycoprotein molecularly controllable-oriented surface imprinted magnetic nanoparticles (GMC-OSIMN) and boric acid functionalized pyrite nanozyme probe (BPNP) to replace the traditional antibody and horseradish peroxidase for sensitive detection of glycoproteins through sandwich detection. In this work, a novel nanozyme functionalized with boric acid was used to label glycoproteins that were captured by GMC-OSIMN. The substrate in the working solution catalyzed by the nanozyme labeled on the protein underwent visible color changes to the naked eye, and the generated signal can be quantitatively detected by a spectrophotometer, and the best color development conditions of the novel nanozyme under the influence of many factors were determined through multi-dimensional investigation. The optimum conditions of sandwich are optimized with ovalbumin (OVA), and it was extended to the detection of transferrin (TRF) and alkaline phosphatase (ALP) in the application. The detection range for TRF was 2.0 × 10-1-1.0 × 104 ng mL-1 with a detection limit of 1.32 × 10-1 ng mL-1, The detection range for ALP was 2.0 × 10-3-1.0 × 102 U L-1 with the detection limit of 1.76 × 10-3 U L-1. This method was subsequently used to detect TRF and ALP levels in 16 liver cancer patients, and the standard deviation of the test results of each patient was less than 5.7%.

Whole-exome sequencing of a novel initiation codon mutation in RUNX2 in a Chinese family with cleidocranial dysplasia.

ABSTRACT: Cleidocranial dysplasia (CCD) is mainly attributable to a variant of runt-related transcription factor 2 (RUNX2) on chromosome 6p21. CCD is an autosomal dominant skeletal disorder characterized by open/delayed closure of fontanels, clavicular hypoplasia, retention of deciduous teeth, and supernumerary permanent teeth. The aim of this study was to investigate potentially pathogenic mutations in 2 Chinese families. Genomic DNA was obtained from peripheral blood lymphocytes, and whole exome sequencing and Sanger sequencing were performed to detect gene variants. Real-time quantitative PCR was performed to determine the mRNA expression level of RUNX2 in the proband of family 1. Silico algorithms and conservation analyses were used to evaluate the functional impact. We identified a novel initiation codon mutation (c.2T>C) and a previously reported mutation (c.569G>A). Familial co-segregation verified an autosomal-dominant inheritance pattern. Our findings demonstrated that the novel mutation c.2T>C causes CCD. Quantitative real-time PCR suggested that downregulated RUNX2 levels and haploinsufficiency in RUNX2 lead to CCD. These results extend the spectrum of RUNX2 mutations in CCD patients and can be used for genetic consultation and prenatal diagnosis.

A novel EDAR missense mutation identified by whole-exome sequencing with non-syndromic tooth agenesis in a Chinese family.

BACKGROUND: Causative variants in genes of the EDA/EDAR/NF-κB pathway, such as EDA and EDARADD, have been widely identified in patients with non-syndromic tooth agenesis (NSTA). However, few cases of NSTA are due to ectodysplasin-A receptor (EDAR) variants. In this study, we investigated NSTA-associated variants in Chinese families. METHODS: Peripheral blood samples were collected from the family members of 24 individuals with NSTA for DNA extraction. The coding region of the EDA gene of the 24 probands was amplified by PCR and sequenced to investigate new variants. Whole-exome sequencing and Sanger sequencing were then performed for probands without EDA variants detected by PCR. RESULTS: A novel missense variant EDAR c.338G>A (p.(Cys113Tyr)) was identified in one family. In addition, three known EDA variants (c.865C>T, c.866G>A, and c.1013C>T) were identified in three families. Genotype-phenotype correlation analysis of EDAR gene mutation showed that NSTA patients were most likely to lose the maxillary lateral incisors and the maxillary central incisors were the least affected. The phenotype of mutations at codon 289 of EDA in NSTA affected patients was characterized by lateral incisors loss, rarely affecting the maxillary first molars. CONCLUSION: A novel EDAR missense variant c.338G>A (p.(Cys113Tyr)) was identified in a family with NSTA, extending the mutation spectrum of the EDAR gene. Genotype-phenotype correlation analyses of EDAR and EDA mutations could help to improve disease status prediction in NSTA families.

Advances and applications of in-tube solid-phase microextraction for analysis of proteins.

In-tube solid-phase microextraction (IT-SPME) with capillary column as extraction device is a well-established green extraction technique with a lot of applications in the fields of biomedicine, food and environment. This article reviews the research contributions of IT-SPME for analysis of proteins. The paper first briefly describes the history of IT-SPME. Then, the development and principle of IT-SPME for analysis of proteins are introduced, in which capillary column configurations of IT-SPME and instruments for quantitative analysis of proteins are summarized. Subsequently, the synthesis strategy and recognition principle of different recognition units, including antibodies, aptamers, molecularly imprinted polymers, and boronate affinity materials, are discussed in detail. This part also introduces several rare recognition units, including lectins, restricted access materials, lysine modified with ß-cyclodextrin and cell membrane. The development trend and possible future direction of IT-SPME for analysis of proteins are mentioned.

A novel EDA1 missense mutation in X-linked hypohidrotic ectodermal dysplasia.

A mutation in the epithelial morphogen gene ectodysplasin-A1 (EDA1) is responsible for the disorder X-linked hypohidrotic ectodermal dysplasia (XLHED), the most common form of ectodermal dysplasia. XLHED is characterized by impaired development of hair, eccrine sweat glands, and teeth. This study aimed to identify potentially pathogenic mutations in four Chinese XLHED families.Genomic DNA was extracted from the peripheral blood and sequenced. Sanger sequencing was used to carry out mutational analysis of the EDA1 gene, and the three-dimensional structure of the novel mutant residues in the EDA trimer was determined. Transcriptional activity of NF-κB was tested by Dual luciferin assay.We identified a novel EDA1 mutation (c.1046C>T) and detected 3 other previously-reported mutations (c.146T>A; c.457C>T; c.467G>A). Our findings demonstrated that novel mutation c.1046C>T (p.A349 V) resulted in XLHED. The novel mutation could cause volume repulsion in the protein due to enlargement of the amino acid side chain. Dual luciferase assay revealed that transcriptional NF-κB activation induced by XLHED EDA1 protein was significantly reduced compared with wild-type EDA1.These results extend the spectrum of EDA1 mutations in XLHED patients and suggest a functional role of the novel mutation in XLHED.

Ethanol production from dilute-acid steam exploded lignocellulosic feedstocks using an isolated multistress-tolerant Pichia kudriavzevii strain.

Renewable and low-cost lignocellulosic wastes have attractive applications in bioethanol production. The yeast Saccharomyces cerevisiae is the most widely used ethanol-producing microbe; however, its fermentation temperature (30-35°C) is not optimum (40-50°C) for enzymatic hydrolysis in the simultaneous saccharification and fermentation (SSF) process. In this study, we successfully performed an SSF process at 42°C from a high solid loading of 20% (w/v) acid-impregnated steam explosion (AISE)-treated rice straw with low inhibitor concentrations (furfural 0.19 g l-1 and acetic acid 0.95 g l-1 ) using an isolate Pichia kudriavzevii SI, where the ethanol titre obtained (33.4 gp  l-1 ) was nearly 39% greater than that produced by conventional S. cerevisiae BCRC20270 at 30°C (24.1 gp  l-1 ). In addition, P. kudriavzevii SI exhibited a high conversion efficiency of > 91% from enzyme-saccharified hydrolysates of AISE-treated plywood chips and sugarcane bagasse, although high concentrations of furaldehydes, such as furfural 1.07-1.21 g l-1 , 5-hydroxymethyl furfural 0.20-0.72 g l-1 and acetic acid 4.80-7.65 g l-1 , were present. This is the first report of ethanol fermentation by P. kudriavzevii using various acid-treated lignocellulosic feedstocks without detoxification or added nutrients. The multistress-tolerant strain SI has greater potential than the conventional S. cerevisiae for use in the cellulosic ethanol industry.

[Establishment of the craniofacial three-dimensional finite element models with the sutures defined alone].

OBJECTIVE: To develop a child craniofacial three-dimensional (3D) finite element model (FEM) with sutures defined alone. METHODS: The CT data for this study was developed from sequential computed tomography scan images taken at 0.625 mm intervals of an 8 years children skull. Data set was imported into Mimics 10.0 and processed with Geomagic 9.0, and exported as initial graphics exchange specification(IGES) files. The IGES files were then imported into Ansys 13.0 to set up two FEM with or without the median palatine suture being opened. The FEM contained nine craniofacial sutures and eight teeth which were defined alone.For simulating orthopedic maxillary protraction, three forces (F1-F2) were loaded on FEM.F1(1 N) was loaded at 1 cm above the geison. F2(1 N) was loaded at articular fossa of temporal bone. F3(2 N) was directed anteriorly and paralleled with occlusal plane near the canine. The stress distribution and the values distributed in each point gained in the two models were compared. RESULTS: Two craniofacial 3D FEM of the child were developed with the median palatine suture opened or not .With median palatine suture being opened or not, the two models showed the similar von Mises stresses (VMS). The distribution of the VMS was in the bridge of the nose and dextro-ala nasi.When the median palatine suture was opened, the max VMS value was 18916.00×10(-4) MPa which appeared in the nose point and the min VMS value was 1.61×10(-4) MPa which appeared in the maxillary central incisor point. At the same time, the max stress value at the direction Y was -3985.30×10(-4) MPa and appeared in the frontomaxillary suture point, and the min Y value was 0.08×10(-4) MPa which appeared in the maxillary central incisor point. When the median palatine suture was not opened, the max VMS value was 19 244.00×10(-4) MPa and appeared in the nose point. The min VMS value was 1.62×10(-4) MPa and appeared in the maxillary central incisor point. At the same time, the max stress value at the direction Y was -4258.20×10(-4) MPa and appeared in the frontomaxillary suture point, and the min Y value was 0.08×10(-4) MPa which appeared in the maxillary central incisor point. CONCLUSIONS: To define the sutures as entities alone contributed to develop child craniofacial 3D FEM which consist nine sutures. There was tiny difference in stress distribution in both the VMS and in Y direction with the median palatine suture being opened or not.