Transcriptome analysis reveals that PbMYB61 and PbMYB308 are involved in the regulation of lignin biosynthesis in pear fruit stone cells.

Pear fruit stone cells have thick walls and are formed by the secondary deposition of lignin in the primary cell wall of thin-walled cells. Their content and size seriously affect fruit characteristics related to edibility. To reveal the regulatory mechanism underlying stone cell formation during pear fruit development and to identify hub genes, we examined the stone cell and lignin contents of 30 'Shannongsu' pear flesh samples and analyzed the transcriptomes of 15 pear flesh samples collected at five developmental stages. On the basis of the RNA-seq data, 35 874 differentially expressed genes were detected. Additionally, two stone cell-related modules were identified according to a WGCNA. A total of 42 lignin-related structural genes were subsequently obtained. Furthermore, nine hub structural genes were identified in the lignin regulatory network. We also identified PbMYB61 and PbMYB308 as candidate transcriptional regulators of stone cell formation after analyzing co-expression networks and phylogenetic relationships. Finally, we experimentally validated and characterized the candidate transcription factors and revealed that PbMYB61 regulates stone cell lignin formation by binding to the AC element in the PbLAC1 promoter to upregulate expression. However, PbMYB308 negatively regulates stone cell lignin synthesis by binding to PbMYB61 to form a dimer that cannot activate PbLAC1 expression. In this study, we explored the lignin synthesis-related functions of MYB family members. The results presented herein are useful for elucidating the complex mechanisms underlying lignin biosynthesis during pear fruit stone cell development.

Study on Improving the Performance of Traditional Medicine Extracts with High Drug Loading Based on Co-spray Drying Technology.

The purpose of this study is to develop modified particles with different structures to improve the flowability and compactibility of Liuwei Dihuang (LWDH) powder using co-spray drying technology, and to investigate the preparation mechanism of modified particles and their modified direct compaction (DC) properties. Moreover, tablets with high drug loading contents were also prepared. Particles were designed using polyvinylpyrrolidone (PVP K30) and hydroxypropyl methylcellulose (HPMC E3) as shell materials, and sodium bicarbonate (NaHCO3) and ammonium bicarbonate (NH4HCO3) as pore-forming agents. The porous particles (Ps), core-shell particles (CPs), and porous core-shell particles (PCPs) were prepared by co-spray drying technology. The key DC properties and texture properties of all the particles were measured and compared. The properties of co-spray drying liquid were also determined and analyzed. According to the results, Ps showed the least improvement in DC properties, followed by CPs, and PCPs showed a significant improvement. The modifier, because of its low surface tension, was wrapped in the outer layer to form a shell, and the pore-forming agent was thermally decomposed to produce pores, forming core-shell, porous, and porous core-shell composite structures. The smooth surface of the shell structure enhances fluidity, while the porous structure allows for greater compaction space, thereby improving DC properties during the compaction process.

Immediate vs Conventional Loading of Mandibular Overdentures: A Comprehensive Systematic Review and Meta-Analysis of Randomized Controlled Trials.

Clinicians treating overdenture patients need to know if immediate loading and conventional loading results in similar outcomes. This study aimed to perform a systematic literature search of studies comparing immediate and conventional loading of mandibular overdentures irrespective of the number of implants and conduct a meta-analysis of implant failure and marginal bone loss (MBL). A literature search of PubMed, ScienceDirect, Ovoid, Springer, and Google Scholar databases was performed for randomized controlled trials (RCTs) comparing immediate vs conventional loading of mandibular overdentures. The primary outcome was implant failure and the secondary outcome was marginal bine loss (MBL). A descriptive analysis was performed for other outcomes. Thirteen trials were included. Only one trial compared the immediate and delayed loading of single implant-supported overdenture. Seven trials used 2 implants, 1 trial used 3 implants while 4 trials used 4 implants. Meta-analysis indicated no statistically significant difference in implant failure and MBL between immediate and conventional loading of 2- and 4-implant supported overdentures. Descriptive analysis indicated no difference in peri-implant tissue indices, implant stability, and quality of life outcomes between the 2 loading protocols. There may be no difference in implant failure and MBL with immediate loading or conventional loading of 2- and 4-implant supported mandibular overdentures. Literature review indicates that there may be no difference in peri-implant tissue indices, implant stability, and quality of life outcomes between the 2 loading protocols. The overall quality of evidence is moderate. Further, adequately powered RCTs are required to strengthen the evidence.

Understanding the Synergistic Correlation between the Spatial Distribution of Drug-Loaded Mixed Micellar Systems and In Vitro Behavior via Experimental and Computational Approaches.

To better promote the application of polymeric mixed micelles (PMMs), a coarse-grained molecular dynamics simulation (CGMD) has been employed to investigate the factors controlling the spatial distribution within the PMMs and predict their drug-loading properties, meanwhile, combined with experimental methods to validate and examine it. In this study, the snapshots obtained from CGMD and the results of proton nuclear magnetic resonance (1H NMR) and transmission electron microscopy (TEM) provide new insights into the distribution principle that the spatial distribution depends on the hydrophobic compatibility of drugs with the regions within PMMs. Docetaxel (DTX) is located within the interior or near the core-corona interface of the HS15 hydrophobic core inside FS/PMMs (PMMs fabricated from a nonionic triblock copolymer (F127)) and a nonionic surfactant (HS15), and therefore, the system with a high HS15 ratio, such as system I, is more suitable for loading DTX. In contrast, the more water-soluble puerarin (PUE) is more likely to be solubilized in the "secondary hydrophobic area," mainly formed by the hydrophobic part of F127 within FS/PMMs. However, when the initial feeding concentration of the drug is increased or the FS mixing ratios are changed, an inappropriate distribution would occur and hence influence the drug-loading stability. Also, this impact was further elucidated by the calculated parameters (solvent-accessible surface area (SASA), the radius of gyration (Rg), and energy landscape), and the analysis of the drug leakage, concluding that inappropriate distribution of the drug would lower the stability of the drug in the PMMs. These results combined together provide new insights into the distribution principle that the spatial distribution of drugs within PMMs depends on the hydrophobic compatibility of drugs with the regions formed by micellar materials. Additionally, in vitro drug release yielded a consistent picture with the above conclusions and provides evidence that both the location of the drug within the systems and the stability of the drug-loading system have a great influence on the drug release behavior. Accordingly, this work demonstrates that we can tune the drug-loading stability and drug release behavior via the drug-PMM interaction and drug location study, and CGMD technology would be a step forward in the search for suitable drug-delivery PMMs.

Membrane fouling in ultrafiltration of natural water after pretreatment to different extents.

The combined fouling during ultrafiltration (UF) of surface water pretreated to different extents was investigated to disclose the roles of polysaccharides, proteins, and inorganic particles in UF membrane fouling. Both reversible and irreversible fouling decreased with enhanced pretreatment (biologically active carbon (BAC) treatment and sand filtration). The sand filter effluent fouled the membrane very slowly. The UF membrane removed turbidity to less than 0.1 nephelometric turbidity unit (NTU), reduced polysaccharides by 25.4%-29.9%, but rejected few proteins. Both polysaccharides and inorganic particles were detected on the fouled membranes, but inorganic particles could be effectively removed by backwashing. The increase of turbidity in the sand filter effluent to 3.05 NTU did not significantly increase the fouling rate, but an increase in the turbidity in the BAC effluent to 6.11 NTU increased the fouling rate by more than 100%. The results demonstrated that the polysaccharide, not the protein, constituents of biopolymers were responsible for membrane fouling. Membrane fouling was closely associated with a small fraction of polysaccharides in the feed water. Inorganic particles exacerbated membrane fouling only when the concentration of fouling-inducing polysaccharides in the feed water was relatively high. The combined fouling was largely reversible, and polysaccharides were the predominant substances responsible for irreversible fouling.

Association between carbohydrate-to-fiber ratio and the risk of periodontitis.

Background/purpose: Periodontitis is a chronic multifactorial inflammatory disease with dental plaque accumulation. This study aimed to analyze the association between carbohydrate-to-fiber ratio and periodontitis risk. Materials and methods: In this cross-sectional study, the data of 6470 participants aged ≥30 years with available oral health exam data of periodontal status were collected from NHANES 2009-2014. Participants were divided into no & mild periodontitis group (n = 3309) and moderate & severe periodontitis group (n = 3161). The possible correlation between the carbohydrate-to-fiber ratio and the risk of periodontitis was explored via univariate and multivariable logistic regression analyses. Odds ratio (OR) and 95% confidence interval (CI) were applied as the effect size. Results: After adjusting for all the confounding factors, carbohydrate-to-fiber ratio <10.89 (OR = 0.82, 95% CI: 0.69-0.97), carbohydrate-to-fiber ratio of 10.89-14.02 (OR = 0.74, 95% CI: 0.63-0.87), carbohydrate-to-fiber ratio of 14.03-18.47 (OR = 0.83, 95% CI: 0.71-0.97) were correlated with reduced risk of periodontitis. In people aged ≥65 years, carbohydrate-to-fiber ratio of 10.89-14.02 (OR = 0.54, 95% CI: 0.30-0.95) and carbohydrate-to-fiber ratio of 14.03-18.47 (OR = 0.56, 95% CI: 0.37-0.86) were correlated with reduced risk of periodontitis in comparison with carbohydrate-to-fiber ratio ≥18.48 group. The decreased risk of periodontitis was also found in males with carbohydrate-to-fiber ratio of 10.89-14.02 (OR = 0.69, 95% CI: 0.49-0.97) and carbohydrate-to-fiber ratio of 14.03-18.47 (OR = 0.73, 95% CI: 0.56-0.95). In people without diabetes, we found that those with carbohydrate-to-fiber ratio <10.89 (OR = 0.75, 95% CI: 0.61-0.93), carbohydrate-to-fiber ratio of 10.89-14.02 (OR = 0.68, 95% CI: 0.53-0.88) and carbohydrate-to-fiber ratio of 14.03-18.47 (OR = 0.84, 95% CI: 0.70-0.99) were linked with lower risk of periodontitis. Conclusion: Low carbohydrate-to-fiber ratio was associated with decreased risk of periodontitis.

Comparative pharmacokinetics of six bioactive components of Shen-Wu-Yi-Shen tablets in normal and chronic renal failure rats based on UPLC-TSQ-MS/MS.

ETHNOPHARMACOLOGICAL RELEVANCE: Shen-Wu-Yi-Shen tablets (SWYST), a Chinese patent medicine consisting of 12 herbal medicines, was formulated by a famous TCM nephrologist, Zou Yunxiang. It is clinically used to improve the symptoms of nausea, vomiting, poor appetite, dry mouth and throat, and dry stool in patients with chronic renal failure (CRF) accompanied by qi and yin deficiency, dampness, and turbidity. SWYST can reduce urea nitrogen, blood creatinine, and urinary protein loss, and increase the endogenous creatinine clearance rate. However, little is known about its pharmacokinetics. AIM OF STUDY: To compare the pharmacokinetics of six bioactive components after oral administration of SWYST in normal and adenine-induced CRF rats. MATERIALS AND METHODS: A method based on ultra-performance liquid chromatography coupled with a triple-stage quadrupole mass spectrometer (UPLC-TSQ-MS/MS) was developed and validated to determine the six bioactive compounds (albiflorin, paeoniflorin, plantagoguanidinic acid, rhein, aloe-emodin, and emodin) in rat plasma. Rat plasma samples were prepared using protein precipitation. Chromatography was performed on an Agilent Eclipse Plus C18 column (3.0 × 50 mm, 1.8 µm) using gradient elution with a mobile phase composed of acetonitrile and water containing 0.1% (v/v) formic acid, while detection was achieved by electrospray ionization MS under the multiple selective reaction monitoring modes. After SWYST administration, rat plasma was collected at different time points, and the pharmacokinetic parameters of six analytes were calculated and analyzed based on the measured plasma concentrations. RESULTS: The UPLC-TSQ-MS/MS method was fully validated for its satisfactory linearity (r ≥ 0.9913), good precisions (RSD <11.5%), and accuracy (RE: -13.4∼13.1%), as well as acceptable limits in the extraction recoveries, matrix effects, and stability (RSD <15%). In normal rats, the six analytes were rapidly absorbed (Tmax ≤ 2 h), and approximately 80% of their total exposure was eliminated within 10 h. Moreover, in normal rats, the AUC0-t and Cmax of albiflorin, plantagoguanidinic acid, and rhein exhibited linear pharmacokinetics within the dose ranges, while that of paeoniflorin is non-linear. However, in CRF rats, the six analytes exhibited reduced elimination and significantly different AUC or Cmax values. These changes may reflect a decreased renal clearance rate or inhibition of drug-metabolizing enzymes and transporters in the liver and gastrointestinal tract caused by CRF. CONCLUSIONS: A sensitive UPLC-TSQ-MS/MS method was validated and used to investigate the pharmacokinetics of SWYST in normal and CRF rats. This is the first study to investigate the pharmacokinetics of SWYST, and our findings elucidate the causes of their different pharmacokinetic behaviors in CRF rats. Furthermore, the results provide useful information to guide further research on the pharmacokinetic-pharmacodynamic correlation and clinical application of SWYST.

Adult dental epithelial stem cell-derived organoids deposit hydroxylapatite biomineral.

Ameloblasts are specialized cells derived from the dental epithelium that produce enamel, a hierarchically structured tissue comprised of highly elongated hydroxylapatite (OHAp) crystallites. The unique function of the epithelial cells synthesizing crystallites and assembling them in a mechanically robust structure is not fully elucidated yet, partly due to limitations with in vitro experimental models. Herein, we demonstrate the ability to generate mineralizing dental epithelial organoids (DEOs) from adult dental epithelial stem cells (aDESCs) isolated from mouse incisor tissues. DEOs expressed ameloblast markers, could be maintained for more than five months (11 passages) in vitro in media containing modulators of Wnt, Egf, Bmp, Fgf and Notch signaling pathways, and were amenable to cryostorage. When transplanted underneath murine kidney capsules, organoids produced OHAp crystallites similar in composition, size, and shape to mineralized dental tissues, including some enamel-like elongated crystals. DEOs are thus a powerful in vitro model to study mineralization process by dental epithelium, which can pave the way to understanding amelogenesis and developing regenerative therapy of enamel.

Soil changes induced by rubber and tea plantation establishment: comparison with tropical rain forest soil in Xishuangbanna, SW China.

Over the past thirty years, Xishuangbanna in Southwestern China has seen dramatic changes in land use where large areas of tropical forest and fallow land have been converted to rubber and tea plantations. In this study we evaluated the effects of land use and slope on soil properties in seven common disturbed and undisturbed land-types. Results indicated that all soils were acidic, with pH values significantly higher in the 3- and 28-year-old rubber plantations. The tropical forests had the lowest bulk densities, especially significantly lower from the top 10 cm of soil, and highest soil organic matter concentrations. Soil moisture content at topsoil was highest in the mature rubber plantation. Soils in the tropical forests and abandoned cultivated land had inorganic N (IN) concentrations approximately equal in NH(4) (+)-N and NO(3) (-)-N. However, soil IN pools were dominated by NH(4) (+)-N in the rubber and tea plantations. This trend suggests that conversion of tropical forest to rubber and tea plantations increases NH(4) (+)-N concentration and decreases NO(3) (-)-N concentration, with the most pronounced effect in plantations that are more frequently fertilized. Soil moisture content, IN, NH(4) (+)-N and NO(3) (-)-N concentrations within all sites were higher in the rainy season than in the dry season. Significant differences in the soil moisture content, and IN, NH(4) (+)-N and NO(3) (-)-N concentration was detected for both land uses and sampling season effects, as well as interactions. Higher concentrations of NH(4) (+)-N were measured at the upper slopes of all sites, but NO(3) (-)-N concentrations were highest at the lower slope in the rubber plantations and lowest at the lower slopes at all other. Thus, the conversion of tropical forests to rubber and tea plantations can have a profound effect on soil NH(4) (+)-N and NO(3) (-)-N concentrations. Options for improved soil management in plantations are discussed.

A multiple-target mRNA-LNP vaccine induces protective immunity against experimental multi-serotype DENV in mice.

Dengue virus (DENV) is a mosquito-borne virus with a rapid spread to humans, causing mild to potentially fatal illness in hundreds of millions of people each year. Due to the large number of serotypes of the virus, there remains an unmet need to develop protective vaccines for a broad spectrum of the virus. Here, we constructed a modified mRNA vaccine containing envelope domain III (E-DIII) and non-structural protein 1 (NS1) coated with lipid nanoparticles. This multi-target vaccine induced a robust antiviral immune response and increased neutralizing antibody titers that blocked all four types of DENV infection in vitro without significant antibody-dependent enhancement (ADE). In addition, there was more bias for Th1 than Th2 in the exact E-DIII and NS1-specific T cell responses after a single injection. Importantly, intramuscular immunization limited DENV transmission in vivo and eliminated vascular leakage. Our findings highlight that chimeric allogeneic structural and non-structural proteins can be effective targets for DENV vaccine and that they can prevent the further development of congenital DENV syndrome.

Enhanced nutrient removal MBR system with chemical addition for low effluent TP.

A pilot study was conducted to test an membrane bioreactor (MBR) process for combined biological and chemical P removal to achieve a very low effluent total phosphorus (TP) concentration of 0.025 mg P/L. With the data from the pilot test, a simulation study was performed to demonstrate that: (1) the pilot system behaviour (effluent quality, MLSS, etc.) can be modelled accurately with an activated sludge model combined with a chemical precipitation model; and (2) with the calibrated model, simulation scenarios can be performed to further understand the pilot MBR process, and provide information for optimizing design and operation when applied at full-scale. Results from the pilot test indicated that the system could achieve very low effluent TP concentration through biological P removal with a limited chemical addition, and chemical addition to remove P to very low level did not affect other biological processes, i.e., organic and nitrogen removal. Simulation studies indicate that the process behaviour can be modelled accurately with an activated sludge model combined with a chemical precipitation model, and the calibrated model can be used to provide information to optimize system design and operation, e.g., chemical addition control under dynamic loading conditions is important for maintaining biological P removal.

[Study on purification of strictosamide from Nauclea officinalis by macroporous resin].

OBJECTIVE: To study on the purification of strictosamide from Nauclea officinalis by macroporous resin to provide reference for production. METHOD: The best macroporous resin was selected among 10 kinds of resins according to adsorption and desorption of the static adsorption experiments. The adsorption quantity, elution volume of water, concentration and elution volume of alcohol were determined according to the single factor experiment. RESULT: HPD400 was the best resin, and the best adsorption quantity was 20.23 mg x g(-1), the elution volume of water and 30% alcohol was 6 BV, and the elution volume of 70% alcohol was 4 BV. CONCLUSION: This technology is simple, feasible, and it can provide reference for the industrialized production.

Metabolomic profiling of ZrO2 nanoparticles in MC3T3-E1 cells.

The authors' previous study showed that zirconium oxide nanoparticles (ZrO2 NPs) induce toxic effects in MC3T3-E1 cells; however, its toxicological mechanism is still unclear. Liquid chromatography-mass spectrometry/time-of-flight mass spectrometry was used to reveal the metabolite profile and toxicological mechanism of MC3T3-E1 cells in response to ZrO2 NPs. The results demonstrated that MC3T3-E1 cells treated with ZrO2 NPs for 24 and 48 h presented different metabolic characteristics. Following ZrO2 NP treatment for 24 h, 96 upregulated and 129 downregulated metabolites in the positive ion mode, as well as 91 upregulated and 326 downregulated metabolites in the negative ion mode were identified. Following ZrO2 NP treatment for 48 h, 33 upregulated and 174 downregulated metabolites were identified in the positive ion mode, whereas 37 upregulated and 302 downregulated metabolites were confirmed in the negative ion mode. Among them, 42 differential metabolites were recognised as potential metabolites contributing to the induced toxic effects of ZrO2 NPs in MC3T3-E1 cells. Most of the differential metabolites were lysophosphatidylcholine and lysophosphatidylethanolamide, indicating that exposure to ZrO2 NPs may have a profound impact on human cellular function by impairing the membrane system. The results also provide new clues for the toxicological mechanism of ZrO2 NP dental materials.

Outcomes of implants placed after osteotome sinus floor elevation without bone grafts: a systematic review and meta-analysis of single-arm studies.

BACKGROUND: The aim of this study is to evaluate the implant survival/success rate, gain in alveolar bone height, crestal bone loss, and complications associated with implants placed in the posterior maxilla after osteotome sinus floor elevation without bone substitutes. METHODS: The electronic databases, such as MEDLINE, EMBASE, CENTRAL, and SCOPUS were systematically and manually searched for publications in peer-reviewed journals. The included articles were subjected to qualitative and quantitative analyses, and the meta-analysis was carried out for single-arm studies. Methodological quality assessment was made for all the included studies. RESULTS: The included studies were of moderate quality, with the overall implant success and survival rates of 98.3% and 97.9% respectively. The most frequent intra-surgical complication was sinus membrane perforation, accounting for 3.08% of the total implants with reported perforations. The overall crestal bone loss in patients with immediate implants placed with OSFE after a 5-year follow-up was 0.957 mm 95%CI (0.538, 1.377). CONCLUSION: Within the limitations of this review, it can be concluded that the survival and success rates of implants placed immediately along with OSFE without any bone substitutes are acceptable and show adequate implant stability with less crestal bone loss over 5 years.

3D­printed Ti6Al4V scaffolds combined with pulse electromagnetic fields enhance osseointegration in osteoporosis.

The loosening and displacement of prostheses after dental implantation and arthroplasty is a substantial medical burden due to the complex correction surgery. Three­dimensional (3D)­printed porous titanium (pTi) alloy scaffolds are characterized by low stiffness, are beneficial to bone ingrowth, and may be used in orthopedic applications. However, for the bio­inert nature between host bone and implants, titanium alloy remains poorly compatible with osseointegration, especially in disease conditions, such as osteoporosis. In the present study, 3D­printed pTi scaffolds with ideal pore size and porosity matching the bone tissue, were combined with pulse electromagnetic fields (PEMF), an exogenous osteogenic induction stimulation, to evaluate osseointegration in osteoporosis. In vitro, external PEMF significantly improved osteoporosis­derived bone marrow mesenchymal stem cell proliferation and osteogenic differentiation on the surface of pTi scaffolds by enhancing the expression of alkaline phosphatase, runt­related transcription factor­2, osteocalcin, and bone morphogenetic protein­2. In vivo, Microcomputed tomography analysis and histological evaluation indicated the external PEMF markedly enhanced bone regeneration and osseointegration. This novel therapeutic strategy has potential to promote osseointegration of dental implants or artificial prostheses for patients with osteoporosis.

Layer-by-layer synthesis of metal-containing conducting polymers: caged metal centers for interlayer charge transport.

Metal-templated [2 + 3]-type cocondensation of a pi-extended boronic acid and nioxime furnished a series of cage molecules, which were electropolymerized to prepare metal-containing conducting polymers (MCPs). Despite sharing essentially isostructural organic scaffolds, these materials display metal-dependent electrochemical properties as evidenced by different redox windows observed for M = Co, Fe, Ru. Consecutive electropolymerization using two different monomers furnished bilayer MCPs having different metals in each layer. In addition to functioning as heavy atom markers in cross-sectional analysis by FIB and EDX, redox-active metal centers participate in voltage-dependent interlayer electron transport to give rise to cyclic voltammograms that are distinctively different from those of each layer alone or random copolymers. A simple electrochemical technique can thus be used as a straightforward diagnostic tool to investigate the structural ordering of electrically conductive layered materials.

Passive medialization of the inferior alveolar nerve to facilitate implant placements in a patient with severe atrophic posterior mandible.

Placing implants in the edentulous jaw of an individual with severe posterior mandibular atrophy is challenging due to the insufficient bone height above the inferior alveolar nerve canal. The different reconstruction techniques available cause extensive morbidity, are costly, and require prolonged treatments. In here we report a case series of patients treated with a new minimally invasive technique to facilitate implant placement in patients with severely atrophied posterior mandible.

Australian Aedes aegypti mosquitoes are susceptible to infection with a highly divergent and sylvatic strain of dengue virus type 2 but are unlikely to transmit it.

BACKGROUND: Humans are the primary hosts of dengue viruses (DENV). However, sylvatic cycles of transmission can occur among non-human primates and human encroachment into forested regions can be a source of emergence of new strains such as the highly divergent and sylvatic strain of DENV2, QML22, recovered from a dengue fever patient returning to Australia from Borneo. The objective of the present study was to evaluate the vector competence of Australian Aedes aegypti mosquitoes for this virus. METHODS: Four- to five-day-old mosquitoes from two strains of Ae. aegypti from Queensland, Australia, were fed a meal of sheep blood containing 108 50% cell culture infectious dose per ml (CCID50/ml) of either QML22 or an epidemic strain of DENV serotype 2 (QML16) isolated from a dengue fever patient in Australia in 2015. Mosquitoes were maintained at 28 °C, 75% relative humidity and sampled 7, 10 and 14 days post-infection (dpi). Live virions in mosquito bodies (abdomen/thorax), legs and wings and saliva expectorates from individual mosquitoes were quantified using a cell culture enzyme-linked immunosorbent assay (CCELISA) to determine infection, dissemination and transmission rates. RESULTS: The infection and dissemination rates of the sylvatic DENV2 strain, QML22, were significantly lower than that for QML16. While the titres of virus in the bodies of mosquitoes infected with either of these viruses were similar, titres in legs and wings were significantly lower in mosquitoes infected with QML22 at most time points although they reached similar levels by 14 dpi. QML16 was detected in 16% (n = 25) and 28% (n = 25) of saliva expectorates at 10 and 14 dpi, respectively. In contrast, no virus was detected in the saliva expectorates of QML22 infected mosquitoes. CONCLUSIONS: Australia urban/peri-urban Ae. aegypti species are susceptible to infection by the sylvatic and highly divergent DENV 2 QML22 but replication of QML22 is attenuated relative to the contemporary strain, QML16. A salivary gland infection or escape barrier may be acting to prevent infection of saliva and would prevent onward transmission of this highly divergent virus in Australia.

[A family survey of syndromes of traditional Chinese medicine in patients with beta-thalassemia].

OBJECTIVE: To explore the relationship between syndromes of traditional Chinese medicine (TCM) and genetic background in patients with beta-thalassemia. METHODS: TCM syndromes were surveyed in the selected 78 patients with beta-thalassemia intermedia including 120 parents. The gene mutations were detected separately. The frequency and score of TCM syndromes between the offspring and their parents in different family types were analyzed, and the differences were compared. RESULTS: The 73 families were divided into two family types by hereditary characteristics. Family type one meant that genotypes of one of the parents were normal, while the offspring genotypes were heterozygous and were exactly the same as another parent. In the 22 families of type one, the heterozygous offspring manifested 6 high-frequency symptoms and signs such as spontaneous perspiration, dry mouth and dry throat, pale or sallow complexion, tidal fever and night sweating, lassitude and pale fingernails. The heterozygous parents manifested 5 high-frequency symptoms and signs such as lassitude in loins and knees, dizziness, aversion to cold and cold limbs, tinnitus, dry mouth and dry throat. The normal parents manifested 3 high-frequency symptoms and signs such as lassitude in loins and knees, dizziness, and spontaneous perspiration. TCM syndrome score in the heterozygous offspring was higher than that in the heterozygous and normal parents, but there was no significant difference (P>0.05). Family type two meant that genotypes of both parents were heterozygous, while the offspring genotypes were heterogenic duplex heterozygotes. In the 51 families of type two, the offspring manifested 9 high-frequency symptoms and signs such as pale or sallow complexion, spontaneous perspiration, dry mouth and dry throat, pale fingernails, tidal fever and night sweating, lassitude, frequent attack of common cold, dysphoria with feverish sensation in chest, and yellow discoloration of the skin and sclera. The parents manifested 3 high-frequency symptoms and signs such as lassitude in loins and knees, dizziness, aversion to cold and cold limbs. TCM syndrome score in the offspring was significant higher than that in the parents (P<0.01). CONCLUSION: In the two family types, TCM syndrome in the offspring is of yin-blood deficiency, while the syndrome of the parents is of kidney deficiency. The differences of TCM syndromes between the offspring and the parents may have some relations to the type of mutant genes and genetically modified ingredients. This research provides scientific evidence to TCM syndrome differentiation treatment of thalassemia.

Synergistic effect of functionalized poly(l-lactide) with surface-modified MgO and chitin whiskers on osteogenesis in vivo and in vitro.

Favorable cytocompatibility and osteogenesis potential are critical for the development of a bone repair material. In this study, two types of surface-modified whiskers, grafted magnesia and chitin (g-MgO and g-CHN) whiskers, were synthesized and introduced into a poly(l-lactide) (PLLA) matrix singly or together to prepare PLLA/g-MgO/g-CHN composite films and bone nails via injection molding. On the account of the synergetic contribution of g-MgO and g-CHN whiskers, the enhanced cell adhesion, spreading, proliferation of mouse embryo osteoblast precursor (MC3T3-E1) cells, as well as the alteration of cell-cycle and inhibition of cell apoptosis, were observed on PLLA/g-MgO/g-CHN film as compared to pure PLLA, PLLA/g-MgO and PLLA/g-CHN films. More importantly, the highest level of the secretion of ALP and the formation of calcium deposition, accompanied with expression of osteogenesis genes (ALP, Runx-2, COL I, OCN) in vitro were obtained for the PLLA/g-MgO/g-CHN film among all of the material groups. Additionally, the PLLA and PLLA composite bone nails were implanted in rabbits' femurs and new bone formation was detected on PLLA/g-MgO/g-CHN group after 16 weeks of implantation by 3D reconstruction of micro-CT and histological analyses. Besides, the bending strength of defected bone repaired by PLLA/g-MgO/g-CHN bone nail was high to 48 MPa, which was far stronger than other bone nail groups. Overall, this study demonstrated the addition of g-MgO and g-CHN whiskers together in PLLA matrix played a synergistic promoting role in cell affinity and osteogenic differentiation, and the developed PLLA/g-MgO/g-CHN composites hold great potential in fields of bone repair.