Pharmacokinetics and safety of pegylated recombinant human granulocyte colony-stimulating factor in children with acute leukaemia.

AIMS: This open-label, phase I study evaluated the pharmacokinetics and safety of pegylated recombinant human granulocyte colony-stimulating factor (PEG-rhG-CSF) for the treatment of chemotherapy-induced neutropenia in children with acute leukaemia. METHODS: PEG-rhG-CSF was administered as a single 100 mcg/kg (3 mg maximum dose) subcutaneous injection at the end of each chemotherapy period when neutropenia occurred. Blood samples were obtained from patients treated with PEG-rhG-CSF. PEG-rhG-CSF serum concentrations were determined by an enzyme-linked immunosorbent assay. Population pharmacokinetic (PPK) analysis was implemented using the nonlinear mixed-effects model. Short-term safety was evaluated through adverse events collection (registered at clinicaltrials.gov identifier: 03844360). RESULTS: A total of 16 acute leukaemia patients (1.8-13.6 years) were included, of whom two (12.5%) had grade 3 neutropenia, six (37.5%) had grade 4 neutropenia, and eight (50.0%) had severe neutropenia. For PPK modelling, 64 PEG-rhG-CSF serum concentrations were obtainable. A one-compartment model with first-order elimination was used for pharmacokinetic data modelling. The current weight was a significant covariate. The median (range) of clearance (CL) and area under the serum concentration-time curve (AUC) were 5.65 (1.49-14.45) mL/h/kg and 16514.75 (6632.45-54423.30) ng·h/mL, respectively. Bone pain, pyrexia, anaphylaxis and nephrotoxicity were not observed. One patient died 13 days after administration, and the objective assessment of causality was that an association with PEG-rhG-CSF was "possible". CONCLUSIONS: The AUC of PEG-rhG-CSF (100 mcg/kg, 3 mg maximum dose) in paediatric patients with acute leukaemia were similar to those of PEG-rhG-CSF (100 mcg/kg) in children with sarcoma. PEG-rhG-CSF is safe, representing an important therapeutic option for chemotherapy-induced neutropenia in paediatric patients with acute leukaemia.

Enhanced anaerobic digestion performance and sludge filterability by membrane microaeration for anaerobic membrane bioreactor application.

Slow dissolution/hydrolysis of insoluble/macromolecular organics and poor sludge filterability restrict the application potential of anaerobic membrane bioreactor (AnMBR). Bubble-free membrane microaeration was firstly proposed to overcome these obstacles in this study. The batch anaerobic digestion tests feeding insoluble starch and soluble peptone with and without microaeration showed that microaeration led to a 65.7-144.8% increase in methane production and increased critical flux of microfiltration membrane via driving the formation of large sludge flocs and the resultant improvement of sludge settleability. The metagenomic and bioinformatic analyses showed that microaeration significantly enriched the functional genes and bacteria for polysaccharide and protein hydrolysis, microaeration showed little negative effects on the functional genes involved in anaerobic metabolisms, and substrate transfer from starch to peptone significantly affected the functional genes and microbial community. This study demonstrates the dual synergism of microaeration to enhance the dissolution/hydrolysis/acidification of insoluble/macromolecular organics and sludge filterability for AnMBR application.

iCVM: An Interpretable Deep Learning Model for CVM Assessment Under Label Uncertainty.

The Cervical Vertebral Maturation (CVM) method aims to determine the craniofacial skeletal maturational stage, which is crucial for orthodontic and orthopedic treatment. In this paper, we explore the potential of deep learning for automatic CVM assessment. In particular, we propose a convolutional neural network named iCVM. Based on the residual network, it is specialized for the challenges unique to the task of CVM assessment. 1) To combat overfitting due to limited data size, multiple dropout layers are utilized. 2) To address the inevitable label ambiguity between adjacent maturational stages, we introduce the concept of label distribution learning in the loss function. Besides, we attempt to analyze the regions important for the prediction of the model by using the Grad-CAM technique. The learned strategy shows surprisingly high consistency with the clinical criteria. This indicates that the decisions made by our model are well interpretable, which is critical in evaluation of growth and development in orthodontics. Moreover, to drive future research in the field, we release a new dataset named CVM-900 along with the paper. It contains the cervical part of 900 lateral cephalograms collected from orthodontic patients of different ages and genders. Experimental results show that the proposed approach achieves superior performance on CVM-900 in terms of various evaluation metrics.

Dental service in 2008 Summer Olympic Games.

OBJECTIVE: How to organise an appropriate team to provide quality dental care during the Olympic Games has become an important consideration for each successive host country. The aims of this study were to document dental services provided at the Olympic Games and to provide data for planning future events. SET-UP OF THE DENTAL CARE DEPARTMENT: There were six dental chairs in six independent treatment rooms, one technical laboratory, a sterilising room and an x-ray room equipped with one digital panoramic screening machine and one intraoral x-ray machine in the polyclinic in the Olympic Village in Beijing. Shifts comprised 80 dentists and 28 nurses who were organised into three shifts working from 08:00 until 23:00. RESULTS: In the 2008 Olympic Games, there were 1607 cases involving 1126 patients requiring dental care: 795 cases from 516 athletes; 483 cases from 370 coaches and other staff; and 99 cases from volunteers. Endodontic treatments, permanent fillings, oral hygiene, mouthguards and treatment of pericoronitis were the most frequent procedures in dental care. The Mouthguard Service was extremely popular and well utilised. 122 athletes received new custom-made mouthguards in Beijing. CONCLUSIONS: As the utilisation of the dental service grows, and the burden of providing care for such a large cohort increases, a well-organised dental team becomes increasingly important. More general-practice dentists were needed in the team. Different kinds of specialist were suggested for working in the team-for example, oral and maxillofacial surgeons, sport dentists for mouthguards and endodontists.

[Analysis on characteristics of 802 emergency cases transported from venues of the Beijing 2008 Olympic Games].

OBJECTIVE: To analyze 802 emergency cases transported from venues of the Beijing 2008 Olympic Games, in order to study the characteristics of critically ill patients and injured in mass gatherings for the purpose of improving medical care and transportation. METHODS: Using the standard medical encounter form system (MEFS) formulated by International Olympic Committee (IOC), the data of patients treated in various venues of Beijing 2008 Olympic Games (2008-07-27 T 09:00/2008-08-27 T 23:00) were analyzed. RESULTS: Eight hundred and two patients had been transported from 227 medical stations of different venues, including 471 male, 331 female, with a mean age of 42.67 years. According to MEFS, 802 patients were divided into 12 types of diseases, i.e. injury (39.03%), digestive disorder (21.82%), cardiovascular disorder (7.36%), respiratory disorder (6.48%), nervous-sensory disorder (6.23%), skin diseases (4.49%), eye disorder (4.24%), ear-nose-throat disorders (3.74%), heat related illness (3.37%), genito-urinary disorder (1.87%), dental disorder (1.37%), psychiatric disorder (0). Patients as grouped according to occupation: workforce (34.04%), non-registered (33.29%), others (13.59%), athletes (12.72%), VIP (3.74%), media (2.62%). CONCLUSION: During a mass gathering the plan for medical support should be formulated with the medical treatment of commonly occurred diseases and high-risk individuals in order to establish an efficient medical emergency transport system.

An Investigation of Softening Laws and Fracture Toughness of Slag-Based Geopolymer Concrete and Mortar.

This paper aimed to determine the softening laws and fracture toughness of slag-based geopolymer (SG) concrete and mortar (SGC and SGM) as compared to those of Portland cement (PC) concrete and mortar (PCC and PCM). Using three-point bending (TPB) tests, the load vs. mid-span displacement, crack mouth opening displacement, and crack tip opening displacement curves (P-d, P-CMOD, and P-CTOD curves) were all recorded. Bilinear softening laws of the PC and SG series were determined by inverse analysis. Furthermore, the cohesive toughness was predicted using an analytical fracture model. The cohesive toughness obtained by experimental study was consistent with that predicted by analytical method, proving the correctness of the tension softening law obtained from inverse analysis. In addition, both initial and unstable fracture toughness values of SG mortar were lower than those of PC mortar given the same compressive strength. Moreover, the initial fracture toughness of SG concrete was generally lower than that of PC concrete, whereas the unstable fracture toughness exhibited an opposite trend.

Mechanical Strength, Surface Properties, Cytocompatibility and Antibacterial Activity of Nano Zinc-Magnesium Silicate/Polyetheretherketone Biocomposites.

In order to improve the biological activity and antibacterial properties of polyetheretherketone (PK) as bone implants, nano zinc-magnesium silicate (nZMS)/PK bioactive composites (nZPC) were fabricated. The results revealed that the mechanical properties, surface roughness and hydrophilicity of the nZPC gradually increased with nZMS content, in which nZPC with 50 w% of nZMS (50nZPC) exhibited the best properties. In addition, incorporation of nZMS into PK significantly improved the apatite mineralization ability of nZPC, which depended on nZMS content. Moreover, the attachment, proliferation and differentiation of MC3T3-E1 cells on nZPC were significantly enhanced with increasing nZMS content. Furthermore, after incorporation of nZMS into PK, the nZPC could inhibit the growth of Escherichia coli (E. coli), in which 50nZPC revealed the best antibacterial activity. The results suggested that 50nZMPC with good bioactivity, cytocompatibility and antibacterial activity might be a promising candidate as an implant for bone repair and anti-infection.

Simultaneous expression of growth hormone releasing hormone (GHRH) and hepatitis B surface antigen/somatostatin (HBsAg/SS) fusion genes in a construct in the skeletal muscle enhances rabbit weight gain.

Somatostatin (SS) and growth hormone-releasing hormone (GHRH) are synthesized and secreted by the hypothalamus, which can control the synthesis and secretion of the growth hormone (GH) from the hypophysis as well as regulate the GH concentrations in animals and humans. In this article, we describe the regulation of animal growth using plasmid DNA encoding both the GHRH gene and the SS gene fused with the hepatitis B surface antigen (HBsAg) gene. We constructed a series of expression plasmids to express the GHRH and HBsAg-SS fusion genes individually as well as collectively. The fusion gene and GHRH were successfully expressed in Chinese hamster ovary (CHO) cells, as proven by reverse transcriptase-polymerase chain reaction (RT-PCR) and immunoblotting tests. Poly D, L-lactide-co-glycolic acid (PLGA) plasmid-encapsulating microspheres were prepared and injected intramuscularly into the leg skeletal muscles of rabbits. Weight gain/day and the levels of insulinlike growth factor-I (IGF-I), SS, and hepatitis B surface antibody (HBsAb) were monitored. During days 30 postinjection, increase in weight gain/day and IGF- I concentration and decrease in SS were observed in treatment groups. From days 15 to 30 postinjection, the weight gain/day significantly increased (P < 0.05) by 129.13%, 106.8%, and 72.82% relative to the control group in the co-expression GHRH and fusion gene (named P-G-HS), fusion gene (named P-HS), and GHRH (named P-G) groups, respectively. And most importantly, the P-G-HS group showed significant weight gain/day (P < 0.05) relative to the P-G and P-HS groups. A significant increase in the IGF-I concentration and decrease in the SS level relative to the control group were also observed. The results indicated that the combination of plasmid-mediated GHRH supplementation and positive immunization against SS led to more robust weight gain/day in rabbits.

Overexpression of GRF encapsulated in PLGA microspheres in animal skeletal muscle induces body weight gain.

Biodegradable nanospheres or microspheres have been widely used as a sustained release system for the delivery of bioagents. In the present study, injectable sustained-release growth hormone-releasing factor (GRF) (1-32) microspheres were prepared by a double emulsion-in liquid evaporation process using biodegradable polylactic-co-glycolic acid (PLGA) as the carrier. The entrapment efficiency was 89.79% and the mean particle size was 4.41 mum. The microspheres were injected into mouse tibialis muscle. After 30 days, mice injected with GRF (1-32) microspheres (group I) gained significantly more weight than any other treatment group, including mice injected with the naked plasmid (group II) (10.26 +/- 0.13 vs. 9.09 +/- 0.56; P < 0.05), a mixture of microspheres and plasmid (group III) (10.26 +/- 0.13 vs. 8.57 +/- 0.02; P < 0.05), or saline (IV) (10.26 +/- 0.13 vs. 6.47 +/- 0.26; P < 0.05). In addition, mice treated with the GRF (1-32) microspheres exhibited the highest expression levels of GRF as detected by PCR, RT-PCR, and ELISA (mean 2.56 +/- 0.40, P < 0.05, overall comparison of treatment with groups II, III, and IV). Additionally, rabbits were injected in the tibialis muscle with the same treatments described above. After 30 days, the group treated with GRF (1-32) microspheres gained the most weight. At day 30 postinjection, weight gain in group I was 63.93% higher than group II (plasmid) (877.10 +/- 24.42 vs. 535.05 +/- 26.38; P < 0.05), 108.59% higher than group III (blank MS) (877.10 +/- 24.42 vs. 420.50 +/- 19.39; P < 0.05), and 93.94% higher than group IV (saline) (877.10 +/- 24.42 vs. 452.25 +/- 27.38; P < 0.05). Furthermore, IGF-1 levels in the serum from GRF microsphere-treated group were elevated relative to all other groups. The present results suggest that encapsulation of GRF with PLGA increases GRF gene expression in muscle after local plasmid delivery, and stimulates significantly more weight gain than delivery of the naked plasmid alone.

[Study on cytomedicine of alginate-poly(L) lysine-alginate microencapsulated hybridoma cells].

AIM: To study the cytomedicine of alginate-poly (L) lysine-alginate (APA) microencapsulated hybridoma cells and their characteristics. METHODS: The spleen cells taken from BALB/C mice immunized with purified human IgG1 kappa type were fused with mouse myeloma cells SP2/0. The hybridoma cell lines secreting monoclonal antibodies (mAb) against human IgG1 kappa type was named JY-A1. The APA microencapsulated JY-A1 cells were prepared with a high-voltage electrostatic system. Microencapsulation parameters were optimized and their morphology was studied. The mechanical strength and chemical intensity of microcapsules were measured. The mAb secrete from APA microencapsulated JY-A1 cells was determined by ELISA kit. The microcapsules injected into mice abdominal cavity previously were recovered at intervals. RESULTS: The microcapsules prepared in the same condition of the high-voltage electrostatic system were round and homogeneous. The mAb secreted by the microencapsulated JY-A1 cells were shown to permeate the membranes of APA microcapsules in vitro. After an intraperitoneal injection to mice, APA microcapsules were recovered on day 7, 14, 28, 56. The electron microscopy study revealed that the majority of recovered microcapsules were intact, and no evidence of immunological reaction in terms of fibrosis. CONCLUSION: APA microencapsulated hybridoma cells prepared by high-voltage electrostatic system have good mechanical strength and chemical intensity. The APA microencapsulated hybridoma cells can maintain physiological functions in vitro, and the microcapsules have good biocompatibility in vivo.

Hepatocyte-targeted psiRNA delivery mediated by galactosylated poly(ethylene glycol)-graft-polyethylenimine in vitro.

Gene silencing in liver disease could be achieved by delivering siRNA with nonviral vectors. However, the transfection efficiency of plasmid siRNA (psiRNA) applied through this approach in hepatocytes is generally low. Based on the fact that the asialoglycoprotein receptors present on hepatocytes can recognize galactose, we synthesized galactosylated poly(ethylene glycol)-graft-polyethylenimine (Gal-PEG-PEI) as a nonviral psiRNA carrier for hepatocyte targeting. Our results indicate that 0.2% (molar percentage) of amine groups of PEI was conjugated with PEG having galactose on its distal end. Increasing the molar ratios of Gal-PEG-PEI to psiRNA in complexation led to a decrease in particle size but an increase in zeta potential of complexes. The transfection efficiency of nanocomplexes, that is, Gal-PEG-PEI/psiRNA, in HepG2 cell line depends on the N/P value, which reflects the molar ratio of Gal-PEG-PEI to psiRNA in the complex. The highest transfection efficiency was 37.34%, which was obtained at N/P 8. At the same N/P value, the transfection efficiency with the nontargeting PEG-PEI/psiRNA or Lipofectamine 2000/psiRNA was much lower. The transfection efficiency of Gal-PEG-PEI/psiRNA dropped to 3.60% from 37.34% after an excessive amount of free galactose was added into the medium for HepG2 cell incubation. By contrast, the similar phenomenon was observed neither when using PEG-PEI or Lipofectamine 2000 as a delivery vector nor in human embryonic kidney 293 cell line lacking ASGR. Real-time PCR analysis and western blot assay demonstrate that the knockdown of HLA-E gene expression by psiRNA/Gal-PEG-PEI (N/P 8) can reach about 60% in HepG2 cells after a 48-h transfection.

Polyethylenimine-grafted copolymer of poly(l-lysine) and poly(ethylene glycol) for gene delivery.

A major challenge in gene therapy is the development of effective gene delivery vectors with low toxicity. In the present study, linear poly(ethylenimine) (lPEI) with low molecular weight was grafted onto the block copolymer (PPL) of poly(l-lysine) (PLL) and poly(ethylene glycol)(PEG), yielding a ternary copolymer PEG-b-PLL-g-lPEI (PPI) for gene delivery. In such molecular design, PLL, lPEI and PEG blocks were expected to render the vector biodegradability, proton buffering capacity, low cationic toxicity and potentially long circulation in vivo, respectively. Given proper control of molecular composition, the copolymers demonstrated lower cytotoxicity, proton buffering capacity, ability to condense pDNA and mediate effective gene transfection in various cell lines. With folate as an exemplary targeting ligand, the FA-PPI/pDNA complex showed much higher transgene activity than its nontargeting counterpart for both reporter and therapeutic genes in folate receptor(FR)-positive cells. FA-PPI mediated effective transfection of the TNF-related apoptosis-inducing ligand gene (TRAIL) in human hepatoma Bel 7402 cells, leading to cell apoptosis and great suppression of cell viability. Our results indicate that the copolymers might be a promising vector combining low cytotoxicity, biodegradability, and high gene transfection efficiency.

[The expression of growth hormone release factor gene carried with PLGA microspheres in muscle tissue and its effect on mouse growth].

Injectable sustained-release pcDNA3-GRF (1-32) microspheres were prepared by double emulsion-in liquid evaporation process,using biodegrable poly lactic-co-glycolic acid as carrier. The enrapment efficiency, mean particle size, drug content thus prepared were 69%, 2.20 microm, 8% and 70% respectively. The result of transfection in vivo showed that after 30 days, accumulative increased body weights on the group injected with pcDNA3-GRF (1-32) microspheres was significantly higher than those group injected with naked plasmid (12.87%), plasmid-empty microspheres (19.72%) and saline (58.58%) respectively. PCR and RT-PCR showed that the expression level of GRF gene on the group injected with pcDNA3-GRF (1-32) microspheres was the highest. GRF gene released by microspheres was still detected after 30 days. In conclusion, pcDNA3-GRF (1-32) microspheres have a controlled release effect and GRF gene could be successfully transfected into muscle cells of mouse by microspheres with higher efficacy and stronger biological function.