Novel construction of multifunctional photo-responsive and nucleic acid-triggered doxorubicin-releasing liposomes for cancer therapy.

All-in-one nano theranostics integrating accurate diagnosis and combined therapy is promising for high-efficacy tumor treatment and receiving significant attention. In this study, we develop photo-controlled release liposomes with nucleic acid-triggered fluorescence and photoactivity for tumor imaging and synergistic antitumor therapy. Copper phthalocyanine as a photothermal agent is fused into lipid layers to prepare liposomes encapsulating cationic zinc phthalocyanine ZnPc(TAP)412+ and doxorubicin, followed by the modification of RGD peptide on the surface to obtain the final product RGD-CuPc:ZnPc(TAP)412+:DOX@LiPOs (RCZDL). RCZDL possesses favorable stability, significant photothermal effect, and photo-controlled release function through the characterization of physicochemical properties. It is shown that the fluorescence and ROS generation could be turned on by intracellular nucleic acid after illumination. RCZDL exhibits synergistic cytotoxicity, increased apoptosis, and significantly promoted cell uptake. Subcellular localization analysis indicates that ZnPc(TAP)412+ tends to be distributed in the mitochondria of HepG2 cells treated with RCZDL after exposure to light. The results of experiments in vivo on H22 tumor-bearing mice demonstrate that RCZDL had excellent tumor targeting, a prominent photothermal effect at the tumor sites, and synergistic antitumor efficiency. More importantly, little RCZDL has been found to be accumulated in the liver, and most were quickly metabolized by the liver. The results confirm that the proposed new intelligent liposomes provide a simple and cost-effective way for tumor imaging and combinatorial anticancer therapy.

Microbiome landscape of lesions and adjacent normal mucosal areas in oral lichen planus patient.

The pathogenesis of oral lichen planus (OLP) remains unclear, and microbial dysbiosis has been proposed to play a role in the pathogenesis of OLP. Oral mucosal swabs from 77 OLP patients and 76 healthy subjects were collected. The bacterial community among the OLP lesion, the adjacent normal mucosal, and the oral mucosal surface in healthy people were analyzed by 16S sequencing. The factor of gender and age that may affect the flora distribution of OLP patients were explored. Results indicate no significant difference in microbiota between OLP and the adjacent group. Compared with the healthy group, Neisseria, Haemophilus, Fusobacterium, Porphyromonas, Rothia, Actinomyces, and Capnocytophaga significantly increased in the OLP group. Actinomyces increased in male OLP patients, and the other six bacteria increased in female OLP patients. In female OLP patients, Lautropia and Dialister were positively correlated with age. While in male OLP patients, Moraxella, Porphyromonas, and Fusobacterium were positively correlated with age. Functional enrichment analysis suggested that abnormal energy metabolism related to ATP synthases, abnormal transport and metabolism of glycans, amino acids, and vitamins, and disorders of the local immune microenvironment might exist in OLP lesion.

Evaluation of culturable 'last-resort' antibiotic resistant pathogens in hospital wastewater and implications on the risks of nosocomial antimicrobial resistance prevalence.

Antimicrobial resistance has been recognized as an important emerging environmental pollutant. 'Last-resort' antibiotics including tigecycline, polymyxin E, daptomycin, vancomycin and linezolid are the 'last line of defence' for antibiotic resistant pathogen infections. Therefore, the presence of 'last-resort' antibiotic resistant pathogens in hospital environments and the nosocomial transmission of 'last-resort' antibiotic resistance poses a grave threat to the well-being of patients. In this work, the extent of resistance to 'last-resort' antibiotics in culturable pathogens in hospital wastewater was investigated. Resistance to 'last-resort' antibiotics were quantified for 1384 culturable Enterobacteriaceae, Enterococcus, Staphylococcus, and Pseudomonas strains. With these investigations, several significant findings were made: (1) a very high level of resistance to 'last-resort' antibiotics was found; (2) multiple resistance to antibiotics, including 'last-resort' antibiotics, was prevalent; (3) a high level of 'last-resort' antibiotic resistance phenotype-genotype inconsistency was found, suggesting knowledge gap for resistance mechanisms; 4) tet(X4)-containing tigecycline-resistant Gram-positive pathogens were found for the first time; 5) wastewater treatment processes are effective in preventing the release of 'last-resort' antibiotic resistant pathogens to the environment. This investigation reveals the severe situation on 'last-resort' resistance in the hospital environment, and implies high risk for nosocomial transmission of 'last-resort' antibiotic resistant pathogens.

Full-state time-varying asymmetric constraint control for non-strict feedback nonlinear systems based on dynamic surface method.

We investigate the tracking control problem for a non-strict feedback nonlinear system with external disturbance and time-varying asymmetric full state constraints. Firstly, the unknown nonlinear term with external disturbance in the system are estimated by fuzzy logic system. The backstepping method is applied to the design of adaptive fuzzy controller. However, to prevent that the constraints are overstepped by introducing an improved log-type time-varying asymmetric barrier Lyapunov function (TABLF) in each step of backstepping design. Secondly, the dynamic surface control (DSC) is introduced in the designed algorithm to solve the computational explosion problem of controller caused by the derivative of control law. The proposed control scheme can speed up the tracking speed of the system. Compared with the previous work, it is verified that the combination of DSC and TABLF can obtain good performance within the constraint range, and can ensure fast and stable tracking convergence under external disturbance. Finally, two simulation examples verify the performance of the adaptive controller.

A lux-based Staphylococcus aureus bioluminescence screening assay for the detection/identification of antibiotics and prediction of antibiotic mechanisms.

The need for the discovery of new antibiotics and solving the antibiotic resistance problem requires rapid detection of antibiotics, identification of known antibiotics, and prediction of antibiotic mechanisms. The bacterial lux genes encode proteins that convert chemical energy into photonic energy and lead to bioluminescence. Exploiting this phenomenon, we constructed a lux-based bioluminescence system in Staphylococcus aureus by expressing lux genes under the control of stress-inducible chaperon promoters. When experiencing antibiotic stress, these constructed reporter strains showed clear bioluminescence response. Therefore, this bioluminescence screening system can be used for the detection of antibiotics in unknown chemical mixtures. Further analysis of bioluminescence response patterns showed that: (1) these bioluminescence response patterns are highly antibiotic specific and therefore can be used for rapid and cheap identification of antibiotics; and that (2) antibiotics having the same mechanism of action have similar bioluminescence patterns and therefore these patterns can be used for the prediction of mechanism for an unknown antibiotic with good sensitivity and specificity. With this bioluminescence screening assay, the discovery and analysis of new antibiotics can be promoted, which benefits in solving the antibiotic resistance problem.

Prognostic ability of DNA-binding protein inhibitor ID-1 expression in patients with oral squamous cell carcinoma.

DNA-binding protein inhibitor ID-1 (ID-1) plays a vital role in the development of cancer. In the present study, ID-1 expression in oral squamous cell carcinoma (OSCC), and its association with prognosis were investigated in 128 patients with OSCC, treated at the Qilu Hospital of Shandong University and followed up for an additional 10 years. Immunohistochemical analysis was performed to detect ID-1 expression, and the association between ID-1 expression and recurrence, and estimated disease-specific survival (DSS) time were subsequently analyzed using the Mann-Whitney U test and the Kaplan-Meier method, respectively. In addition, the log-rank test was implemented to compare the survival curves and multivariate Cox proportional hazards analysis was performed to assess the prognostic value of ID-1. The results demonstrated that ID-1 was highly expressed in the majority of OSCC tissues investigated, and ID-1 expression was significantly higher in cases with recurrence of local tumors and lymph node metastasis. Furthermore, higher ID-1 expression levels were associated with a shorter DSS time. Taken together, the results of the present study suggest that ID-1 may serve as an independent prognostic factor to predict DSS time in patients with OSCC.

Comparison of microbiomes in ulcerative and normal mucosa of recurrent aphthous stomatitis (RAS)-affected patients.

BACKGROUND: Recurrent aphthous stomatitis (RAS) is the most common form of oral ulcerative disease, whose cause is still unknown. Researchers have found the association of many factors with the occurrence of RAS, and proposed oral bacterial infection could be a cause for this disease. METHODS: To investigate whether the occurrence of RAS is associated with oral bacterial infection, we performed high throughput sequencing analysis of bacterial samples collected from the normal oral mucosa and aphthous ulcers of 24 patients. RESULTS: Firmicutes, Proteobacteria and Bacteriodetes were the most abundant phyla in the microbiomes analysed. The alpha diversities of the oral mucosa and aphthous ulcer microbiomes were similar, suggesting a similar richness and diversity. The NMDS analysis showed the oral mucosa and aphthous ulcer microbiomes are significantly different. This suggestion is further supported by Anosim, MRPP, and Adonis analyses. More detailed comparison of the two groups of microbiomes suggested that the occurrence of RAS is significantly associated with the increase of Escherichia coli and Alloprevotella, as well as the decrease of Streptococcus. CONCLUSIONS: Considering E. coli is a very common intestinal bacterium, we propose that E. coli colonization could be a cause for RAS, and controlling E. coli colonization could help curing RAS.

Enhanced aerobic granulation at low temperature by stepwise increasing of salinity.

High salinity and low temperature are generally considered to have negative effects on the formation, stability and performance of aerobic granular sludge (AGS). This study investigated whether and how salinity acclimation strategies can enhance aerobic granulation at low temperature (12 °C) in three sequencing batch reactors (SBRs). Stepwise increased concentrations of NaCl (2-10 and 4-20 g/L) were added to the influent of R1 and R2 with steps of 1 and 2 g/L per week respectively, while R0 was set as a control (salt-free). The granulation processes in R1 and R2 were rapidly started up within 9 days, and were completed within 21 and 18 days, respectively. By contrast, R0 took 25 days and 49 days to start and complete granulation. The salinity acclimation strategies improved sludge hydrophobicity, reduced repulsion barrier between cells, and stimulated EPS production during granulation processes, which simultaneously promoted the formation of AGS. When the influent salinity reached 14 g/L on day 35, granule hydrophobicity, density and size in R2 sharply decreased and granules began to disintegrate afterwards. When operated under salt-free condition, sludge bulking occurred in R0 since day 60. The treatment performance was thus impaired in these two reactors, especially in R2 with significant biomass loss. Conversely, the AGS developed in R1 maintained stable structure with high biomass concentration (8.0 gSS/L) and excellent treatment performance for COD (90%), ammonium (95%) and total nitrogen (70%). Genera Thauera, Azoarcus, and Nitrosomonas were more enriched, while Flavobacterium and Meganema were more suppressed in R1, which would have contributed to granule stability and treatment performance. In conclusion, great care has to be taken for cultivating and operating AGS at low temperature for treating saline wastewater. Increasing salinity with a lower salt gradient provides a possibility for rapid granulation of AGS with excellent treatment performance under such conditions.

The Genetic Structures of an Extensively Drug Resistant (XDR) Klebsiella pneumoniae and Its Plasmids.

Multi-, extensively-, and pan-drug resistant bacteria are a threat to our health today, because their wide resistance spectra make their infections difficult to cure. In this work, we isolated an extensively drug resistant (XDR) Klebsiella pneumoniae 2-1 strain from the stool sample of a patient diagnosed of colorectal cancer. K. pneumoniae 2-1 was found to be resistant to all the antibiotics tested except for cefepime, tigecycline, and ceftazidime-avibactam. By sequencing the complete genome of K. pneumoniae 2-1, we found it contains a chromosome of 5.23 Mb and two circular plasmids with the size of 246 and 90 kb. The larger plasmid, pKP21HI1 was found to be a new conjugation-defective plasmid belonging to incompatibility group HI1B and a new sequence type. Further comparative genomics analysis and antimicrobial resistance gene analysis showed that although a great deal of changes took place on the chromosome of K. pneumoniae 2-1 in comparison with the reference genome, the extensively drug resistance phenotype of K. pneumoniae 2-1 is primarily due to the two multidrug resistant plasmids it contains. This work explains the genetic and mechanistic basis of the extensive drug resistance of K. pneumoniae 2-1, and found that plasmids play key roles in the strong antibiotic resistance of bacteria.

Magnetic Resonance Imaging (MRI) Evaluation for Anterior Disc Displacement of the Temporomandibular Joint.

BACKGROUND Magnetic resonance imaging (MRI) is the criterion standard imaging technique for visualization of the temporomandibular joint (TMJ) region, and is currently considered the optimum modality for comprehensive evaluation in patients with temporomandibular joint disorder (TMD). This study was aimed at finding the value of MRI in pre-clinical diagnosis of TMJ disc displacement. MATERIAL AND METHODS Patients primarily diagnosed as having anterior disc displacement by clinical symptoms and X-ray were selected in the present study. MRI was used to evaluate surrounding anatomical structures and position, as well as morphological and signal intensity change between patients and normal controls. RESULTS Posterior band position was significantly different between the patient group and control group. At the maximum opened-mouth position, the location of disc intermediate zone returned to normal. At closed-mouth position, the thickness of anterior and middle, but not posterior, band increased. The motion range of the condyle in the anterior disc displacement without reduction (ADDWR) patient group was significantly less than the value in the anterior disc displacement with reduction (ADDR) patient group and the control group. Whether at closed-mouth position or maximum opened-mouth position, the exudate volume in the patient group was greater than in the normal group. CONCLUSIONS MRI can be successfully used to evaluate multiple morphological changes at different mouth positions of normal volunteers and patients. The disc-condyle relationship can serve as an important indicator in assessing anterior disc displacement, and can be used to distinguish disc displacement with or without reduction.

Striking changes in volatile profiles at sub-zero temperatures during over-ripening of 'Beibinghong' grapes in Northeastern China.

The evolution of free and glycosidically bound volatile compounds in 'Beibinghong' (Vitis vinifera×Vitis amurensis) grape berries throughout on-vine over-ripening and freezing processes was studied in two vintages. The aroma profiles of 'Beibinghong' icewine berries were characterized by C6 compounds, higher alcohols and terpenoids in free fractions and carbonyl compounds, higher alcohols, C6 alcohols and terpenoids in bound fractions. With regard to free volatile compounds, there was a decrease in the concentration of C6 compounds, terpenols and norisoprenoids and an increase of terpene oxides during over-ripening process. A striking alteration of volatile profile occurred at sub-zero temperatures, particularly for the free fractions such as C6 alcohols, higher alcohols and oxidative terpene derivatives. These changes were attributed to a series of reactions (biotransformation, oxidation and anaerobic metabolism) induced by water loss and especially freeze-thaw cycles. PCA revealed temperature and rainfall affected the accumulation of volatile compounds during over-ripening processes.

Study on angle of immediate loading of immediate implant placement.

OBJECTIVE: To investigate the clinical immediate load at an angle after immediate placement of the implant. METHODS: Select 4 adult dogs; through establishing the angle loading animal experiment model, perform lateral loading on 32 implants respectively at vertical and 0°, 10°, and 20°, with which as a basis for grouping, determine the osseointegration index and new bone growth rate; and observe the peri-implant bone remodeling conditions. RESULTS: The 20° group is found with the most obvious bone absorption, and compared with other groups, its osseointegration index and new bone growth rate are statistically significant (P < 0.01); bone remodeling under 0° load stress is the best, with the formation of new bone and the highest bone contact ratio, which is the most reasonable under this the stress distribution compared with other angles. CONCLUSIONS: The implant stress distribution at 0° against the occlusal force direction is closer to physiologic optimum stress on the implant bone interface, and it is permitted for the long axis of the immediately implanted and immediately loaded implant to be tilted within about 10° against the load angle.

Magnetic resonance imaging of temporomandibular joint: morphometric study of asymptomatic volunteers.

BACKGROUND: The aims of this study were to determine the best suited magnetic resonance imaging scanning plane, scanning sequence, and imaging modality for the evaluation of the temporomandibular joint (TMJ) and quantitatively assess the relationship of articular disk position to condyle position. METHODS: One hundred four TMJs in 52 symptom-free heads were examined by magnetic resonance imaging. The best scanning plane, scanning sequence, and scanning parameter were determined according to the imaging time and image quality. Bilateral symmetry of the articular disk and mandibular condyle was measured by using the automatic measurement of 3.0-T GE Excite Signa MR scanner. RESULTS: Fast spin-echo sequence, oblique sagittal imaging plane, and proton density imaging were the best suited scanning sequence, scanning planes, and imaging modality, respectively. The thicknesses of the anterior and posterior bands and for the intermediate zone were not statistically different for both sides. The posterior band of the disk was found to originate in an area adjacent to the 12-o'clock position of the condyle (± 5 degrees), whereas the anterior band of the disk originated adjacent to 1-o'clock position (28 ± 6 degrees). The anteroposterior diameter and mediolateral diameter of the condylar processes were not statistically different for both sides. The axial condylar angle between the plane of the greatest mediolateral diameter of the condylar processes and the midsagittal plane were also not statistically different for both sides. CONCLUSIONS: The magnetic resonance images can depict clearly major regional anatomic structures and position in the TMJ, which can be used in the early diagnosis for the TMJ disorder.

Adipose-derived stem cells transfected with pEGFP-OSX enhance bone formation during distraction osteogenesis.

This study was designed to investigate the effects of local delivery of adipose-derived stem cells (ADSCs) transfected with transcription factor osterix (OSX) on bone formation during distraction osteogenesis. New Zealand white rabbits (n=54) were randomly divided into three groups (18 rabbits per group). A directed cloning technique was used for the construction of recombinant plasmid pEGFP-OSX, where EGFP is the enhanced green fluorescence protein. After osteodistraction of the right mandible of all experimental rabbits, rabbits in group A were treated with ADSCs transfected with pEGFP-OSX, group B with ADSCs transfected with pEGFP-N1, and group C with physiological saline. Radiographic and histological examinations were processed after half of the animals within each group were humanely killed by injection of sodium pentothal at Week 2 or 6 after surgery. The distraction bone density was measured as its projectional bone mineral density (BMD). Three parameters were measured, namely, the thickness of new trabeculae (TNT), and the volumes of the newly generated cortical bone (NBV1) and the cancellous bone (NBV2) of the distracted regions. Good bone generation in the distraction areas was found in group A, which had the highest BMD, TNT, and NBV in the distraction zones among the groups. There was no significant difference in bone generation in the distraction areas between groups B and C. The results indicate that the transplantation of ADSCs transfected with pEGFP-OSX can effectively promote bone generation during distraction in vivo.

Transcription factor osterix modified bone marrow mesenchymal stem cells enhance callus formation during distraction osteogenesis.

This study was designed to investigate the effects of local delivery of bone marrow mesenchymal stem cells (BMMSCs) with or without osterix (OSX) gene transfected on bone regeneration in the distracted zone using a rabbit model of mandibular lengthening. Fifty-four New Zealand white rabbits underwent osteodistraction of the left mandible and were then randomly divided into group A, group B, and group C (n = 18 for each group). At the end of distraction BMMSCs transfected with OSX, autologous BMMSCs and physiological saline were injected into the distraction gaps in groups A, B, and C, respectively. Nine animals from each group were humanely killed at 2 and 6 weeks after completion of distraction. The distracted mandibles were harvested and processed for radiographic, histological, and immunohistochemical examination. Excellent bone formation in the distracted callus was observed in group A and group B; the former showed better bone formation and highest bone mineral density (BMD), thickness of new trabeculae (TNT, mm) and volumes of the newly formed bone area (NBV) in the distraction zones. Group C animals showed poor bone formation in the distracted callus when compared with groups A and B. Positive immunostaining of bone sialoprotein (BSP) was observed in the distracted callus in all groups; however, BSP expression was much stronger in group A than in groups B and C. The results of this study suggest transplantation of BMMSCs can promote bone formation in DO; OSX-mediated ex vivo gene therapy was more effective during bone deposition and callus formation in distraction osteogenesis.

Transarterial embolization of a high-flow maxillary arteriovenous malformation fed by multiple arteries.

A high-flow maxillary arteriovenous malformation fed by multiple arteries, including bilateral internal maxillary arteries and ophthalmic artery, is reported. A combination of polyvinyl alcohol particles and N-butyl-cyanoacrylate mixed with lipiodol was progressively deposited within the distal vascular bed by transarterial superselective embolization of the branches of bilateral internal maxillary arteries, resulting in complete anatomic and clinical cure. The authors feel that this approach is more secure and effective compared with transfemoral venous embolization and direct transosseous puncture. It also avoids mutilating surgery.

[Relationship between transforming growth factor-alpha gene polymorphism and non-syndromic cleft lip with cleft palate].

OBJECTIVE: To study the association of TGF-alpha gene polymorphism and non-syndromic cleft lip with cleft palate in Shandong province. METHODS: Polymerase chain reaction combined with restrict enzyme digestion was used to detect the target gene variation in 98 patients with non-syndromic cleft lip with cleft palate and 101 healthy controls. RESULTS: The C2 allele frequency in patients with non-syndromic cleft lip with cleft palate was significantly higher than that in healthy controls. The genotype frequency in patients with positive family history was significantly higher than that without positive family history. CONCLUSION: TGF-alpha gene polymorphism is closely associated with non-syndromic cleft lip with cleft palate in Shandong, especially in patients with positive family history.