Study on the preparation and feasibility of a novel adding-type biological slow-release carbon source.

The development of slow-release carbon sources is an effective biological treatment to remove nutrients from wastewater with low carbon-to-nitrogen ratio (C/N). Most filling-type slow-release carbon could not fulfil the needs of current wastewater treatment plants (WWTPs) process. And most adding-type slow-release carbon sources were prepared using some expensive chemical materials. In this study, combining the advantages of the aforementioned types, a novel adding-type wastepaper-flora (AT-WF) slow-release carbon source was proposed, aiming to realise wastepaper recycling in WWTPs. The screening and identification of the mixed flora, AT-WF carbon source release behaviour, and denitrification performance were investigated. The results showed that through the proposed screening method, a considerable proportion of cellulose-degradation-related genera was enriched, and the cellulose degradation ability and ratio of readily available carbon sources of flora T4, S4 and S5 were effectively strengthened. AT-WF had significant carbon release ability and stability, with an average total organic carbon (TOC) release of 8.82 ± 2.36 mg/g. Kinetic analysis showed that the entire carbon release process was more consistent with the first-order equation. Piecewise fitting with the Ritger-Peppas equation exhibited that the rapid-release (RR) stage was skeleton dissolution and the slow-release (SR) stage was Fick diffusion. Denitrification efficiency can achieve a high average removal efficiency of 94.17%, which could theoretically contribute 11.2% more to the total inorganic nitrogen (TIN) removal. Thus, this study indicated that AT-WF could be utilised as an alternative carbon source in WWTPs.

Pretreatment quality of life as a predictor of survival for patients with nasopharyngeal carcinoma treated with IMRT.

BACKGROUND: To evaluate the prognostic significance of pretreatment quality of life for patients with nasopharyngeal carcinoma treated with intensity-modulated radiotherapy. METHODS: We performed a prospective, longitudinal study on 554 newly diagnosed patients with NPC from April 2011 to January 2015. A total of 501 consecutive NPC patients were included. Patients were asked to complete the EORTC QLQ-C30 (version 3.0) and QLQ-H&N35 questionnaires before treatment. RESULTS: Global health status among QLQ-C30 correlates with EBV DNA(P = 0.019). In addition, pretreatment appetite loss was significantly correlated with EBV DNA(P = 0.02). Pretreatment teeth, opening mouth, feeding tube was significantly correlated with EBV DNA, with P value of 0.003, < 0.0001, and 0.031, respectively. In multivariate analysis, pretreatment cognitive functioning of QLQ-C30 was significantly associated with LRFS, with HR of 0.971(95%CI 0.951-0.990), P = 0.004. Among scales of QLQ-H&N35 for multivariate analysis, pretreatment teeth (P = 0.026) and felt ill (P = 0.012) was significantly associated with PFS, with HR of 0.984 (95%CI 0.971-.998) and 1.004 (95%CI 1.001-1.007), respectively. Felt ill of QLQ-H&N35 was significantly associated with DMFS, with HR of 1.004(95%CI 1.000-1.007), P = 0.043. There is no QoL scale significantly associated with OS after multivariate analysis. CONCLUSIONS: In conclusion, our analysis confirms that pretreatment teeth and felt ill was significantly associated with PFS in NPC patients treated with IMRT. In addition, the posttreatment EBV DNA was significantly associated with OS.

Evaluation of Food Fineness by the Bionic Tongue Distributed Mechanical Testing Device.

In this study, to obtain a texture perception that is closer to the human sense, we designed eight bionic tongue indenters based on the law of the physiology of mandibular movements and tongue movements features, set up a bionic tongue distributed mechanical testing device, performed in vitro simulations to obtain the distributed mechanical information over the tongue surface, and preliminarily constructed a food fineness perception evaluation model. By capturing a large number of tongue movements during chewing, we analyzed and simulated four representative tongue movement states including the tiled state, sunken state, raised state, and overturned state of the tongue. By analyzing curvature parameters and the Gauss curvature of the tongue surface, we selected the regional circle of interest. With that, eight bionic tongue indenters with different curvatures over the tongue surface were designed. Together with an arrayed film pressure sensor, we set up a bionic tongue distributed mechanical testing device, which was used to do contact pressure experiments on three kinds of cookies-WZ Cookie, ZL Cookie and JSL Cookie-with different fineness texture characteristics. Based on the distributed mechanical information perceived by the surface of the bionic tongue indenter, we established a food fineness perception evaluation model by defining three indicators, including gradient, stress change rate and areal density. The correlation between the sensory assessment and model result was analyzed. The results showed that the average values of correlation coefficients among the three kinds of food with the eight bionic tongue indenters reached 0.887, 0.865, and 0.870, respectively, that is, a significant correlation was achieved. The results illustrate that the food fineness perception evaluation model is effective, and the bionic tongue distributed mechanical testing device has a good practical significance for obtaining food texture mouthfeel information.

Calcium Channel Opening Rather than the Release of ATP Causes the Apoptosis of Osteoblasts Induced by Overloaded Mechanical Stimulation.

BACKGROUND: Stress fracture is one of the most common overuse injuries in athletes. Overloaded mechanical stimulation is an important factor affecting stress fractures, but the mechanism is unclear. METHODS: MC3T3-E1 cells and a polycaprolactone (PCL) scaffold were co-cultured, and finite element analysis (FEA) was used to analyze the load-carrying capability. Cell proliferation was investigated with CCK-8 assays. An alkaline phosphatase (AKP) activity assay was used to evaluate cell differentiation. Cell apoptosis was analyzed using Hoechst/ PI double-labeling, Caspase-3 activity and lactate dehydrogenase (LDH) activity assays. Realtime PCR and Western blotting were used to examine the gene and protein expression, respectively, of Caspase-3 and Caspase-9. Assays of the intracellular calcium with fluorescent probe technique and extracellular ATP with fluorometric assay kit were used to analyze the changes in the intracellular calcium concentration induced by calcium channel opening and the release of ATP, respectively, at different operation times. RESULTS: When the apparent strain reached 10000 µÎµ, the strain scope of fber at levels greater than 4000 µÎµ was 60%. Overloading for 4 days and operation times of 0.5 h and 2 h increased the cell number and AKP secretion. However, apoptosis genes were activated at the same time, and the operation time of 2 h had a significantly greater effect than 0.5 h. At 8 days, the cell numbers were greater for the operation time of 0.5 h than for 2 h, and the 2-h groups had the fastest apoptosis rate. Overloading for 1 day increased intracellular calcium levels and ATP release. The increase in intracellular calcium could be blocked by the addition of N-ethylmaleimide (NEM) or Hank's medium. Overloading for 8 days increased intracellular calcium levels but decreased extracellular ATP, and verapamil blocked the increase in intracellular calcium. CONCLUSION: We found that a simultaneous 'double effect' on osteoblasts was induced by overloading, which promoted cell proliferation, differentiation and apoptosis. Short-term overloading could open the cell membrane calcium channels and release calcium stores to elevate intracellular calcium levels, thereby promoting the proliferation and differentiation of cells to a greater extent than the effect of apoptosis. For long-term overloading, calcium channel opening in the membrane could lead to overloading of intracellular calcium levels, inducing an apoptosis effect that is greater than the effect on proliferation and differentiation.

TLR3 signaling in macrophages is indispensable for the protective immunity of invariant natural killer T cells against enterovirus 71 infection.

Enterovirus 71 (EV71) is the most virulent pathogen among enteroviruses that cause hand, foot and mouth disease in children but rarely in adults. The mechanisms that determine the age-dependent susceptibility remain largely unclear. Here, we found that the paucity of invariant natural killer T (iNKT) cells together with immaturity of the immune system was related to the susceptibility of neonatal mice to EV71 infection. iNKT cells were crucial antiviral effector cells to protect young mice from EV71 infection before their adaptive immune systems were fully mature. EV71 infection led to activation of iNKT cells depending on signaling through TLR3 but not other TLRs. Surprisingly, iNKT cell activation during EV71 infection required TLR3 signaling in macrophages, but not in dendritic cells (DCs). Mechanistically, interleukin (IL)-12 and endogenous CD1d-restricted antigens were both required for full activation of iNKT cells. Furthermore, CD1d-deficiency led to dramatically increased viral loads in central nervous system and more severe disease in EV71-infected mice. Altogether, our results suggest that iNKT cells may be involved in controlling EV71 infection in children when their adaptive immune systems are not fully developed, and also imply that iNKT cells might be an intervention target for treating EV71-infected patients.

Type I Interferons Triggered through the Toll-Like Receptor 3-TRIF Pathway Control Coxsackievirus A16 Infection in Young Mice.

UNLABELLED: Coxsackievirus A16 (CVA16) is one of the major etiological agents of hand, foot, and mouth disease (HFMD) in children. The host defense mechanisms against CVA16 infection remain almost entirely unknown. Unlike previous observations with enterovirus 71 (EV71) infection, here we show that gamma interferon (IFN-γ) or invariant NK T cell deficiency does not affect disease development or the survival of CVA16-infected mice. In contrast, type I interferon receptor deficiency resulted in the development of more severe disease in mice, and the mice had a lower survival rate than wild-type mice. Similarly, a deficiency of Toll-like receptor 3 (TLR3) and TRIF, but not other pattern recognition receptors, led to the decreased survival of CVA16-infected mice. TLR3-TRIF signaling was indispensable for the induction of type I interferons during CVA16 infection in mice and protected young mice from disease caused by the infection. In particular, TRIF-mediated immunity was critical for preventing CVA16 replication in the neuronal system before disease occurred. IFN-ß treatment was also found to compensate for TRIF deficiency in mice and decreased the disease severity in and mortality of CVA16-infected mice. Altogether, type I interferons induced by TLR3-TRIF signaling mediate protective immunity against CVA16 infection. These findings may shed light on therapeutic strategies to combat HFMD caused by CVA16 infection. IMPORTANCE: Hand, foot, and mouth disease (HFMD) is a major threat to public health in the Asia-Pacific region. Both CVA16 and EV71 are major pathogens that are responsible for HFMD. The majority of research efforts have focused on the more virulent EV71, but little has been done with CVA16. Thus far, host immune responses to CVA16 infection have not yet been elucidated. The present study discovered an initial molecular mechanism underlying host protective immunity against CVA16 infection, providing the first explanation for why CVA16 and EV71 cause different clinical outcomes upon infection of humans. Therefore, different therapeutic strategies should be developed to treat HFMD cases caused by these two viruses.

Repairing cartilage defects with bone marrow mesenchymal stem cells induced by CDMP and TGF-ß1.

The aim of this study was to explore the ability for chondrogenic differentiation of bone marrow mesenchymal stems cells (BMSCs) induced by either cartilage-derived morphogenetic protein 1 (CDMP-1) alone or in the presence of transforming growth factor-ß1 (TGF-ß1) in vivo and in vitro. BMSCs and poly-lactic acid/glycolic acid copolymer (PLGA) scaffold were analyzed for chondrogenic capacity induced by CDMP-1 and TGF-ß1 in vivo and in vitro. Chondrogenic differentiation of BMSCs into chondrocytes using a high density pellet culture system was tested, whether they could be maintained in 3-D PLGA scaffold instead of pellet culture remains to be explored. Under the culture of high-density cell suspension and PLGA frame, BMSCs were observed the ability to repair cartilage defects by either CDMP-1 alone or in the presence of TGF-ß1 in vitro. Then the cell-scaffold complex was implanted into animals for 4 and 8 weeks for in vivo test. The content of collagen type II and proteoglycan appeared to increase over time in the constructs of the induced groups (CDMP in the presence of TGF-ß1), CDMP group and TGF group. However, the construct of the control group did not express them during the whole culture time. At 4 and 8 weeks, the collagen type II expression of the induced group was higher than the sum of TGF group and CDMP group by SSPS17.0 analysis. BMSCs and PLGA complex induced by CDMP-1 and TGF- ß1 can repair cartilage defects more effectively than that induced by CDMP-1 or TGF-ß1 only.

Pretreatment with Daudi cells eliminates anti-CD47 monoclonal antibody interference in immunohematology testing.

BACKGROUND: Anti-CD47 monoclonal antibodies have increasing clinical applications in the treatment of cancer. However, anti-CD47 monoclonal antibodies interfere with immunohematology testing in patients who require blood transfusion. As the current approaches to removing any interferences have technical problems, new methods need to be developed to resolve anti-CD47 interference in immunohematology testing. MATERIALS AND METHODS: We evaluated the Daudi cell line for the adsorption of free anti-CD47 monoclonal antibodies from patients' plasma to facilitate immunohematology testing in patients treated with anti-CD47 monoclonal antibody. CD47 expression was identified on the Daudi cells using flow cytometry and confocal microscopy. Next, we tested the ability of intact Daudi cells mixed with simulating plasma and clinical samples to achieve efficient removal of interfering anti-CD47 monoclonal antibodies. The indirect antiglobulin test was used to verify whether interference from anti-CD47 monoclonal antibodies in plasma was eliminated and whether the detection of other irregular antibodies was affected. The effect of eliminating interference was also investigated in relation to the time that the Daudi cells were stored after having been fixed with paraformaldehyde. RESULTS: CD47 expression was higher on Daudi cells than on red blood cells. Analysis of the indirect antiglobulin test results revealed that anti-CD47 monoclonal antibody-treated patients' plasma absorbed by Daudi cells for 15 min at 37°C could completely prevent the interference of anti-CD47 monoclonal antibodies in immunohematology testing while the detection of the tested antibodies, including anti-D and anti-K, was unaffected. DISCUSSION: By decreasing the incubation time, we discovered that interferences in samples with agglutination strengths below 2+ could be eliminated after incubation for 5 min. Of importance, Daudi cells can be preserved with 4% paraformaldehyde for 14 days as short-term storage reagents. This is the first study in which Daudi cells were used to effectively resolve the interference of anti-CD47 monoclonal antibodies in pretransfusion tests.

Low-intensity pulsed ultrasound reduces alveolar bone resorption during orthodontic treatment via Lamin A/C-Yes-associated protein axis in stem cells.

BACKGROUND: The bone remodeling during orthodontic treatment for malocclusion often requires a long duration of around two to three years, which also may lead to some complications such as alveolar bone resorption or tooth root resorption. Low-intensity pulsed ultrasound (LIPUS), a noninvasive physical therapy, has been shown to promote bone fracture healing. It is also reported that LIPUS could reduce the duration of orthodontic treatment; however, how LIPUS regulates the bone metabolism during the orthodontic treatment process is still unclear. AIM: To investigate the effects of LIPUS on bone remodeling in an orthodontic tooth movement (OTM) model and explore the underlying mechanisms. METHODS: A rat model of OTM was established, and alveolar bone remodeling and tooth movement rate were evaluated via micro-computed tomography and staining of tissue sections. In vitro, human bone marrow mesenchymal stem cells (hBMSCs) were isolated to detect their osteogenic differentiation potential under compression and LIPUS stimulation by quantitative reverse transcription-polymerase chain reaction, Western blot, alkaline phosphatase (ALP) staining, and Alizarin red staining. The expression of Yes-associated protein (YAP1), the actin cytoskeleton, and the Lamin A/C nucleoskeleton were detected with or without YAP1 small interfering RNA (siRNA) application via immunofluorescence. RESULTS: The force treatment inhibited the osteogenic differentiation potential of hBMSCs; moreover, the expression of osteogenesis markers, such as type 1 collagen (COL1), runt-related transcription factor 2, ALP, and osteocalcin (OCN), decreased. LIPUS could rescue the osteogenic differentiation of hBMSCs with increased expression of osteogenic marker inhibited by force. Mechanically, the expression of LaminA/C, F-actin, and YAP1 was downregulated after force treatment, which could be rescued by LIPUS. Moreover, the osteogenic differentiation of hBMSCs increased by LIPUS could be attenuated by YAP siRNA treatment. Consistently, LIPUS increased alveolar bone density and decreased vertical bone absorption in vivo. The decreased expression of COL1, OCN, and YAP1 on the compression side of the alveolar bone was partially rescued by LIPUS. CONCLUSION: LIPUS can accelerate tooth movement and reduce alveolar bone resorption by modulating the cytoskeleton-Lamin A/C-YAP axis, which may be a promising strategy to reduce the orthodontic treatment process.

Denitrification performance and in-situ fermentation mechanism of the wastepaper-flora slow-release carbon source.

Using wastepaper as external carbon sources is an optional way to achieve total nitrogen removal faced with low carbon to nitrogen ratio municipal sewage. Most of studies have primarily focused on using cellulose-rich wastes establishing the separate denitrification units to achieve in-situ fermentation, which can cause blockages and prolong the process chain. In response, a novel in-situ fermentation wastepaper-flora slow-release carbon source (IF-WF) was proposed using in the original denitrification unit. IF-WF could be efficiently utilized in situ and the denitrification rate increased with the increase of nitrate nitrogen. The fermentation products were highly available, but internal acidification of IF-WF inhibited fermentation. Moreover, IF-WF limited the growth of polysaccharides in the extracellular polymeric substances of denitrified sludge. IF-WF finally formed the structure dominated by nitrate-reduction bacteria outside and cellulose-degrading bacteria inside. These results provide guidance for understanding the mechanism of IF-WF for in-situ fermentation to promote nitrogen removal.

[Experimental research on the compatibility of self-assembly nanofiber hydrogel from the amphipathic peptide containing IKVAV with olfactory ensheathing cells of rats].

The present research was aimed to explore the biocompatibility of IKVAV self-assembling peptide nanofiber scaffold with olfactory ensheathing cells (OECs) of rats. The OECs were seeded onto the surface of coverslips covered with IKVAV self-assembling peptide nanofiber scaffold hydrogel (2D culture system), and implanted within IKVAV self-assembling peptide nanofiber scaffold hydrogel (3D culture system), respectively. The adhesion, viability of OECs were observed with inverted microscope. Then the characteristics for survival and adhesion of cells by image processing were observed, and statistical analysis on the number of S-100 positive cell, the area of the cell bodies and the perimeter of the cell and MTT method were carried out. It was found that the OECs could survive and migrate in IKVAV self-assembling peptide nanofiber scaffold. The result of the cell MTT exam, of the shape and quantity of cells had no significant difference compared to those of the OECs cultured with poly-L-lysine (PLL). It has been proved that IKVAV self-assembling peptide nanofiber scaffold has good biocompatibility with rat OECs.

Longitudinal detection of somatic mutations in saliva and plasma for the surveillance of oral squamous cell carcinomas.

PURPOSES: Although clinical and radiological examinations can be used to diagnose oral cancer, and surgical pathology remains the gold standard, these conventional methods have limitations. We evaluated the feasibility of longitudinal next-generation sequencing-based liquid biopsy for oral squamous cell carcinoma surveillance. MATERIALS AND METHODS: Eleven patients were enrolled, and plasma and saliva were collected before, and 1, 3, and 6 months after surgery. Tumor-specific mutations were selected using paired, whole-exome analyses of tumor tissues and whole blood. Genes frequently mutated in head and neck cancer were identified using the Cancer Genome Atlas (TCGA) and Catalogue of Somatic Mutations in Cancer (COSMIC) databases to design targeted deep sequencing panels. RESULTS: In five of the six patients with recurrent cancer, circulating tumor DNA (ctDNA) was detected earlier with liquid biopsy than with conventional monitoring techniques. Moreover, patients without recurrence exhibited decreased ctDNA allele frequency post-treatment. CONCLUSIONS: Longitudinal liquid biopsy of plasma and saliva may be feasible for detecting somatic mutations associated with oral squamous cell carcinomas. It might be attributable to determine early tumor recurrence through genetic analysis of ctDNA.

Proanthocyanidins: Components, Pharmacokinetics and Biomedical Properties.

Proanthocyanidins (PAs) are a group of polyphenols enriched in plant and human food. In recent decades, epidemiological studies have upheld the direct relationship between PA consumption and health benefits; therefore, studies on PAs have become a research hotspot. Although the oral bioavailability of PAs is quite low, pharmacokinetics data revealed that some small molecules and colonic microbial metabolites of PAs could be absorbed and exert their health beneficial effects. The pharmacological effects of PAs mainly include anti-oxidant, anticancer, anti-inflammation, antimicrobial, cardiovascular protection, neuroprotection, and metabolism-regulation behaviors. Moreover, current toxicological studies show that PAs have no observable toxicity to humans. This review summarizes the resources, extraction, structures, pharmacokinetics, pharmacology, and toxicology of PAs and discusses the limitations of current studies. Areas for further research are also proposed.

F-containing initiatior for ultrasensitive fluorescent detection of lung cancer DNA via atom transfer radical polymerization.

An ultrasensitive fluorescence method for early diagnosis of lung cancer via Nafion-initiated atom transfer radical polymerization (ATRP) is reported, in this paper. In the proposed method, thiolated peptide nucleic acid (PNA) is modified to amino magnetic beads (MBs) via a cross-linking agent to specifically capture target DNA (tDNA), and the initiator (Nafion) of ATRP is attached to PNA/DNA heteroduplexes based on the phosphate groups of the tDNA and sulfonate groups of Nafion via phosphate-Zr4+-sulfonate chemistry. Nafion as a macroinitiator of ATRP possesses multiple C-F active sites to initiate polymerization, and numerous polymeric chains that significantly amplify the fluorescent signal are formed. Under optimal conditions, a good linear relationship is obtained in the range of 0.1 nM-0.1 fM with correlation coefficients of 0.9975, and the detection limit is as low as 35.5 aM (∼214 molecules). The proposed strategy has several advantages of simplicity, cost-effectiveness, selectivity and sensitivity. More importantly, the anti-interference results demonstrate that the proposed Nafion-initiated ATRP strategy has great potential in bioanalytical applications.

Granular activated carbon assisted nitrate-dependent anaerobic methane oxidation-membrane bioreactor: Strengthening effect and mechanisms.

Eutrophication and global warming are two main urgent environmental problems around the world. Nitrate-dependent Anaerobic Methane Oxidation (NdAMO) is a bioprocess coupling nitrate reduction with anaerobic methane oxidation, which could mitigate of these two environmental issues simultaneously. In this study, a newly granular active carbon-NdAMO-membrane bioreactor (GAC-NdAMO-MBR) system was established to evaluate its nitrogen removal efficiency, membrane fouling property and the probable strengthening mechanism was also uncovered. Results indicated that the nitrate removal rate in GAC-NdAMO-MBR reached 31.85 ± 3.19 mgN·L-1·d-1 while it was only 10.35 ± 2.02 mgN·L-1·d-1 in NdAMO-MBR system (lack of GAC), which was multiplied three-fold. The membrane flux decay rate of GAC- NdAMO -MBR was 0.15 L/m2·h·d while it was 0.49 L/m2·h·d without GAC, and the addition of GAC could extend membrane fouling time for 2.5 times. Notablely, the relative abundance of NdAMO bacteria sharply increased from 27.15% to 56.91% after GAC addition while the NdAMO archaea showed similar variation trend. The physicochemical property of GAC mainly contributed the strengthening effect. The porous structure of GAC absorbed methane and adhered by microorganism, which enhance microorganism amount and metabolic activity. The mechanical strength of GAC scoured membrane surface to mitigate external fouling and pores absorbed EPS to reduce internal fouling. The combined effects could improve NdAMO microorganism growth and metabolism activity and finally improved nitrogen removal performance and controlled membrane fouling. These findings could deep the knowledge of NdAMO process and help extend its application potential in environment science and engineering.

A novel RNA-splicing mutation in COL1A1 gene causing osteogenesis imperfecta type I in a Chinese family.

BACKGROUND: Osteogenesis imperfecta (OI), also known as brittle bone disease, is a rare heterogeneous group of inherited disorders characterized by low bone mass and increased bone fragility. The four major clinical criteria for diagnosis of OI are osteoporosis with abnormal fragility of the skeleton, blue sclera, dentinogenesis imperfecta, and premature otosclerosis. The presence of two of these abnormalities confirms the diagnosis. More than 90% patients have autosomal dominant mutations in one of the two genes, COL1A1 and COL1A2, that encode the alpha chains of type I collagen. While the diagnosis of OI is still based on clinical and radiological grounds, there is a growing demand for the molecular characterization of causative mutations. Although there have been several studies on the mutational spectra of COL1A1 and/or COL1A2 in Western populations, very few cases have been reported from Asia. The purpose of this study is to report two patients with OI type I in a Chinese family, who had a novel RNA-splicing mutation in COL1A1 gene and describe the molecular, radiological and clinical findings. METHODS: The proband, (case II-5), a 32-y-old Chinese male, and his 7-y-old daughter were diagnosed as OI type I according to their clinical and radiological features. Genomic DNA was extracted from their blood samples and all promoters, exons and exon/intron boundaries of COL1A1 and COL1A2 genes were sequenced. Polymerase chain reaction sequence-specific primers (PCR-SSP) was used to confirm patients' heterozygous state. RESULTS: Direct DNA sequencing analysis of COL1A1 gene revealed a splicing mutation (c.1875+1G>A, also as IVS 27+1G>A) that converted the 5' end of intron 27 from GT to AT. This mutation was found in both 2 affected individuals but 9 unaffected relatives and the 50 controls were not observed, which was consistent with the clinical diagnosis. This mutation (c.1875+1G>A) appeared to be novel, which is neither reported in literature nor registered in the Database of Collagen Mutations. The heterozygous states of patients' intron 27 were confirmed by PCR-SSP. CONCLUSION: We identify a novel RNA-splicing mutation (c.1875+1G>A) in COL1A1 gene resulting in OI type I in a Chinese family. The detailed molecular and clinical features will be useful for extending the evidence for genetic and phenotypic heterogeneity in OI and exploring the phenotype-genotype correlations in OI.

Facial soft tissue changes after nonsurgical rapid maxillary expansion: a systematic review and meta-analysis.

BACKGROUND: The present systematic review and meta-analysis aimed to test the hypothesis that no facial soft tissue changes occur after nonsurgical rapid maxillary expansion (RME), in order to provide a reference for orthodontists. METHODS: PubMed, EMBASE, Cochrane Library, OVID, MEDLINE, CINAHL, Scopus, and ScienceDirect databases were electronically and manually searched up to December 2017, and randomized controlled, clinical controlled trials, cohort studies and retrospective studies where soft tissue changes were measured before and after nonsurgical RME were identified. Study appraisal and synthesis were performed by two reviewers who completed the study selection and quality assessment procedures independently and in duplicate. Data from the involved studies were pooled using Revman 5.3. RESULTS: A total of 1762 articles were identified after the removal of duplicates. After selection and quality assessment, 15 studies met the inclusion criteria for the systematic review, and 13 articles were ultimately included in the meta-analysis. The quality of the involved studies was relatively moderate. Pre-expansion, postexpansion, and postretention data were pooled. The nasal width, alar base width, and distances from the lower lips to the E line showed significant changes after expansion. Moreover, after retention, the nasal width, mouth width, upper philtrum width, and distance from the lower lip to the E line showed significant increases relative to the baseline values. Limitations of the present study included the moderate quality of the included studies and the fact that the results were based on short-term observations of patients in the growth phase. CONCLUSION: Our findings suggest that RME results in a significantly increased nasal width, mouth width, upper philtrum width, and distance from the lower lip to the E line after the retention phase. However, the clinical importance of these findings is questionable.

Effects of fixed orthodontic brackets on oral malodor: A systematic review and meta-analysis according to the preferred reporting items for systematic reviews and meta-analyses guidelines.

BACKGROUND: This systematic review and meta-analysis aimed to identify whether there is any relationship between fixed orthodontic appliances and malodor, and if self-ligating brackets (SLBs) prevent malodor better than conventional brackets (CBs). METHODS: The electronic databases PubMed, Ovid, EMBASE, and the Cochrane Library were searched from inception to September 2016; a manual search was also performed. Randomized controlled and clinical controlled trials, in which experimental groups received fixed orthodontic therapy and malodor was measured, were included. Patients treated with fixed orthodontic brackets were compared with those without any treatment, and SLB systems were compared with CB systems. Two reviewers independently selected potentially relevant studies, evaluated the risk for bias, extracted essential data, and synthesized findings using Review Manager version 5.3 (Copenhagen: The. Nordic Cochrane Centre, The Cochrane Collaboration, 2014). RESULTS: Four studies, involving a total of 152 participants, met the inclusion criteria. Fixed orthodontic appliances caused malodor from the initial visit to 2 to 3 months, but was only significant after the first week (mean difference 20.24 [95% confidence interval [CI]11.75-28.74]; P < .00001). Plaque index, gingival index, and periodontal pocket depths demonstrated no statistical differences between the SLB and CB groups after the first week. However, SLBs significantly controlled malodor better than CBs after the first week (mean difference 4.32 [95% CI 6.02 to 2.61]; P < .00001). The quality of the included studies was relatively low and relevant research in this field is quite scarce. CONCLUSIONS: Although the evidence base was relatively weak, fixed orthodontic treatment appeared to be a risk factor for malodor, independent of periodontal changes, and SLB systems controlled malodor better than CB systems.

Individual construction of freeform-fabricated polycaprolactone scaffolds for osteogenesis.

The construction of engineered bone mostly focuses on simulating the extracellular matrix (ECM) for proper biological activity. However, the complexity of architecture and the variability of the mechanical properties of natural bones are related to individual differences in age, nutritional state, mechanical loading and disease status. Defect substitutions should be normed with the host natural bone, balancing architectural and mechanical adaption, as well as biological activity. Using a freeform fabrication (FFF) method, we prepared polycaprolactone (PCL) scaffolds with different architectures. With simulation of structural and mechanical parameters of rabbit femur cancellous bone, individual defect substitution with the characteristics of the rabbit femur was obtained with high porosity and connectivity. Biological adaption in vitro was examined and osteoid formation in vivo was assessed by implantation in situ. Simulating the femur cancellous bone, 300-µm FFF PCL scaffolds had better architectural and mechanical properties. The protocol produced an architecturally, mechanically and biologically adaptive construction of an individual model for rapid-prototype PCL scaffolds. A guide system was developed to accurately reproduce virtually individual defect substitutions of the bone.

Functional diversity of family 3 ß-glucosidases from thermophilic cellulolytic fungus Humicola insolens Y1.

The fungus Humicola insolens is one of the most powerful decomposers of crystalline cellulose. However, studies on the ß-glucosidases from this fungus remain insufficient, especially on glycosyl hydrolase family 3 enzymes. In the present study, we analyzed the functional diversity of three distant family 3 ß-glucosidases from Humicola insolens strain Y1, which belonged to different evolutionary clades, by heterogeneous expression in Pichia pastoris strain GS115. The recombinant enzymes shared similar enzymatic properties including thermophilic and neutral optima (50-60 °C and pH 5.5-6.0) and high glucose tolerance, but differed in substrate specificities and kinetics. HiBgl3B was solely active towards aryl ß-glucosides while HiBgl3A and HiBgl3C showed broad substrate specificities including both disaccharides and aryl ß-glucosides. Of the three enzymes, HiBgl3C exhibited the highest specific activity (158.8 U/mg on pNPG and 56.4 U/mg on cellobiose) and catalytic efficiency and had the capacity to promote cellulose degradation. Substitutions of three key residues Ile48, Ile278 and Thr484 of HiBgl3B to the corresponding residues of HiBgl3A conferred the enzyme activity towards sophorose, and vice versa. This study reveals the functional diversity of GH3 ß-glucosidases as well as the key residues in recognizing +1 subsite of different substrates.