Model-informed drug development (MIDD) in the development of pediatric drugs is drawing more and more attention due to the insufficiency of subjects, lack of research on ontogeny, and the limitation of ethic. The core of MIDD used for dose selection includes the population pharmacokinetic (PopPK) model and physiologically based pharmacokinetic (PBPK) model, as well as model-based simulation and prediction. PBPK model has the advantage of predicting the optimal pediatric dose before the clinical trials and has the ability of extrapolation from adult model to pediatric model. PopPK model characterizes the pediatric PK feature based on the analysis of clinical data and can be used to explore the significant covariates, which is a power tool for individualized medicine in children. With their own advantages and disadvantages, PBPK and PopPK model should be jointly used in the pediatric drug development to refine the dose regimen for children at different ages. In this study, the pediatric drug development of rivaroxaban was taken as an example to introduce the combined application of PBPK model and PopPK model in the design and validation of pediatric dose regimen in Phase Ⅰ, Ⅱ and Ⅲ trials, which may provide reference to MIDD in other pediatric drug development.

Objective To compare the agreement among Chinese 1992, 2008 and UICC 2010 staging systems of nasopharyngeal carcinoma (NPC) and evaluate their predictive value of radiotherapeutic prognosis.Methods 347 NPC patients without distant metastasis treated in our hospital from 2000 to 2005 were retrospectively analyzed.Every patient was categorized into T, N, and clinical stage by Chinese 1992, 2008 and UICC 2010 staging systems, respectively.Kappa value was used to evaluate the agreement among three systems.Kaplan-Meier method was used to analyze the 5-year overall survival (OS), local-free survival (LFS) and distant metastasis-free survival (DMFS), the difference between subgroup was tested by Logrank.Results The agreement of clinical stage, T and N stage between Chinese 2008 and UICC 2010 staging system was better than that of them compared to 1992 staging system, Kappa value were 0.700、0.881 and 0.722.The agreement of T stage was better than N and clinical stage among these three staging system.The difference of OS between stageⅢ and stage Ⅳ was significant in Chinese 2008 and UICC 2010 staging system (χ2=4.48,P=0.034;χ2=8.88,P=0.003), and with no different in 1992 staging system (χ2=0.40,P=0.526).There was no significant difference of LFS between T1 and T2,T2 and T3,T3 and T4 in all staging systems (χ2=1.85,0.53,0.50,P=0.174,0.467,0.479;χ2=1.25,2.10,1.99,P=0.264,0.148,0.159;χ2=0.77,0.60,0.87, P=0.381,0.441,0.350).There were no significant differencesin 1992 staging system, while there was significant differences of DMFS between N1 and N2, N2 and N3 in 2008 stage system, N1 and N2 in UICC 2010 stage system.Conclusions The predictive value of Chinese 2008 and UICC 2010 staging system for prognosis were similar, and were better than that of 1992 staging system in NPC.

Objective:To evaluate the accuracy of the application of color Doppler sonography (CDS) and computer tomography angiography (CTA) in preoperative perforator identification and flap design and provide theoretical support for the restoration of oral maxillofacial defect with free superficial circumflex iliac artery perforator flap (SCIAPF). Methods: (1) Preoperative CDS and CTA techniques were performed to map the SCIA perforators of 29 adult patients diagnosed with malignant tumor in the oral maxillofacial head and neck regions. These patients were scheduled for concurrent reconstruction surgery. (2) A diagnostic test was designed to com-pare the CDS and CTA techniques. Results:(1) A total of 18 patients underwent flap preparation. SCIA was not found in one of the pa-tients during surgery, but was observed intra-operatively in the other 17 patients. The average SCIA diameter was 0.69 ± 0.20 mm. (2) The diagnostic test showed a CDS sensitivity of 75.0%, a CDS specificity of 82.4%, and an area under the ROC curve of 0.79. The CTA sensitivity was 75.0%, the specificity was 94.2%, and the area under the ROC curve was 0.85. The diameters measured by CDS and CTA were compared with the diameter measured intra-operatively. Significant differences were observed among the three diame-ters (P<0.05). The average diameter measured by CDS was 0.84 ± 0.14 mm. The average diameter measured by CTA was 1.01 ± 0.19 mm. Conclusion:CDS and CTA are relatively reliable technologies for preoperative detection of perforator vessel. The use of CDS and CTA technology mapping for SCIAPF can provide accurate information about the perforator, including the position of the perforator and the relationship between the peripheral tissues and the caliber of the vessel.

Objective:To study the expression of EG-1 in thyroid cancer tissue and analyze its correlation with clinical prognosis. Methods:EG-1 mRNA levels in malignant thyroid tissues were evaluated by RT-PCR. All patients were followed up. The survival rates were analyze in EG-1 positive group and EG-1 negative group. Benign thyroid tissues were used as control. Results:EG-1 was expressed in thyroid cancer. The differences between expression of malignant thyroid tissues and benign thyroid tissues were statistically significant (P

Objective To study the relationship between pedometer-based daily physical activity and obesity measurements in adult male leaders living in countryside.Methods A total of 136 men who received routine health screening were measured for body weight,body mass index (BMI),waist circumstance,and total body fat.Daily walking activities were determined by pedometer for 7 days.Results Ⅰ to Ⅱ grade walking activities was found in 58% participants.Daily physical activities were negatively correlated with body weight,waist circumference,BMI and total body fat.Body weight,waist circumference,BMI and total body fat were almost normal when walking activities increased to Ⅲ grade.Only 7% of the subjects participated in exercises.Conclusion Pedometer-based daily physical activities may be significantly correlated with obesity arameters.Increased walking activities will help to loss weight.

Objective To discuss the therapeutic options for treatment of subaxial cervical dislocation combined with facet locking. Methods There were 49 patients with cervical dislocations including 7 patients with dislocation at C3,4, 15 at C4,5, 14 at C5,6 and 13 at C6,7. Eleven patients were with old dislocation, with duration of dislocation ranging from 2 hours to 61 days. Neurologic status of the patients according to Frankel scale was graded A in 14 patients, grade B in nine, grade C in 10 and grade D in nine. All patients were treated surgically after closed reduction with skull traction. Results The successful reduction rate was 63% for fresh dislocation, with average improvement of 0.65 grade for spinal cord function. All bone grafts got fusion at four months after operation. Conclusion Therapeutic options are based on fresh or old dislocations, paraplegia or not, intervertebral disk injury severity, and reduction or not through traction for patients with lower cervical dislocations.

Objective To investigate the influence of murine cytomegalovirus(MCMV) infection on differentiation and differentiation gene expression of neural stem cells (NSCs) in vitro for studying the mechanisms of brain abnormalities calmed by congenital cytomegalovirns infection. Methods NSCs were separated from fetal BALB/c mouse and cultured and identified in vitro. The differentiation potency of NSCs was observed by immunnfluorescence. The NSCs infected by MCMV at dosage of multiplicity of infection (MOI) equaled to 5, I and 0. 1, respectively, were cultured in differentiation medium. The morphological changes of the cells were observed by inverted microscope. The ratios of NSCs and its differentiated cells were detected by flow cytometry. The expression changes of nestin, GFAP and NSE, markers of NSCs and its differentiated cells, were studied by immunofluorescence ( MOI = 1 ). The expression of early antigen (EA) of MCMV was detected to observe the infection process. Real-time RT-PCR method was employed to measure the expression levels of the key differentiation genes Wnt-3 and Wnt-7a in Wnt signal pathway of NSCs at early phage of differentiation culture. Results NSCs isolated from embryonic mouse brains could proliferate to form neurnspheres and strongly express Nestin and differentiate into NF-200 positive neurons or GFAP positive astrocytes. The NSCs of the infected groups couldn't adhere to the wall and appear differentia-tion growth, but showed swollen gradually after differentiation culture. The nostin expression of the infected groups downregulated slowly and was higher than that of the control groups ( P < 0.05 ). The GFAP and NSE expression of the infected groups were lower than that of the control groups (P <0.05). The EA of MCMV could be always detected in the cells of the infected groups. The ratios of nestin positive cells of the infected groups were higher than that of the control groups, but the ratios of GFAP and NSE positive cells of the for-mer were lower than that of the latter from 3rd to 9th day after differentiation culture ( P < 0.05 ). The levels of Wnt-3 mRNA and Wnt-7a mRNA of the infected groups were markedly lower than that of the control groups from 1st to 2nd clay and from 12th hour to 2nd day after differentiation culture respectively ( P < 0.05 ) . These changes of the infected groups became more obvious as MCMV MOI increased . Conclusion MCMV could inhibit significantly NSCs differentiate to neurons and astrocytes and lead to the decrease of dif-ferentiated cells. MCMV could inhibit or interfere with the gene expression of Wnt-3 and Wnt-7a in Wnt sig-nal pathway of NSCs. The effect that MCMV inhibited the differentiation and the differentiation gene expres-sion of NSCs showed dose-dependent with MCMV MOI. The inhibitory effect of MCMV on the differentiation of NSCs might be induced by interfering the differentiation gene expression of NSCs, which is possibly the one of primary causes of brain development disorders caused by congenital CMV infection.

OBJECTIVETo assess the feasibility of a polymer material-expanded polytetrafluoroethylene (ePTFE) as wound covering.

METHODSForty-five Sprague Dawley (SD) rats inflicted with open wound on the back were divided into 3 groups, each group 15 rats. Wounds in the autograft group were covered with autologous skin. Wounds in the allograft group were covered with skin from 15 Wistar rats (donor). Wounds in the experiment group were covered with ePTFE. Wound healing in each group was observed with naked eye. Tissue specimens collected on 3rd, 7th, and 14th post-operative day (POD) respectively were stained with HE and examined under light microscope. Macrophages, lymphocytes and fibroblasts were quantified. IL-2 and IFN-gamma levels in the infiltrating mononuclear cells were assessed by immunohistochemistry with expression of gray value.

RESULTSWounds in rats of autograft group and experiment group healed well without redness, swelling or infection. Rejection was evident in the allograft group rats on 8th POD, including epidermis degeneration and focal necrosis in different degree, plus red swelling on wound edge. The numbers of macrophages, fibroblasts, lymphocytes in allograft group were significantly higher than those in autograft group and experiment group (P < 0.01). The average gray values of IL-2 and IFN-gamma of allograft group on 7th POD were 129 +/- 7 and 113.7 +/- 2.7, which were significantly lower than those in the experiment group (144 +/- 8, 137.3 +/- 1.9, respectively, P < 0.01) and allograft group (189 +/- 6, 180.3 +/- 3.7, respectively, P < 0.01).

CONCLUSIONSePTFE will induce minimal inflammatory response and foreign body reaction. It is unlikely to cause side effect when applied to wound surface for a prolonged period. Thus, it is suitable to use ePTFE as wound surface covering.

Animals ; Bandages ; Graft Rejection ; Male ; Polytetrafluoroethylene ; therapeutic use ; Rats ; Rats, Sprague-Dawley ; Skin Transplantation ; Tissue Engineering ; Transplantation, Homologous ; Wound Healing

Human cytomegalovirus (HCMV) is a DNA virus and serious opportunistic pathogen for both newborn and immunocompromised individuals.To research technique for gene silence and antiviral agents, ribozyme M1GS-T6 was constructed from external guide sequences(EGSs)that consist of a sequence complementary to HCMV UL54 gene RNA and M1 RNA, the catalytic RNA subunit of RNase P from Escherichia coli. The results showed that M1GS can efficiently cleave the mRNA sequence encoding UL54 protein in vitro.

To study the effect of endotoxin on liver apoptosis, L02 liver cells were cultured and passaged in vitro, and then stimulated by endotoxin at 10 mg/mL for 4, 8, 16 and 24 h respectively. Liver apoptosis was flow cytometrically and fluorescently detected. Immunohistochemistry was used to detect the delivery of smac and caspase9. The delivery of liver cell smac and the activity of caspase3 were measured by caspase3 assay kit. The hepatic failure models of rats were established by using D-galactosamine. The blood serum and liver tissues were collected for the detection of the liver function, the level of endotoxin and the activity of caspase3 by using chromogenic substrate limulus amebocyte lysate method (LAL) and caspase3 active assay kit. The expression of smac and caspase9 in liver cells was detected by Western blotting. With in vitro study, the L02 cells stimulated by LPS condensed into conglobation and formed apoptotic bodies. After those cells were stained by hoechst, the apoptotic cells displayed blue color under the fluorescent microscope. The apoptosis rate was increased over time and the apoptosis was mainly of advanced stage. Meanwhile, the rate of smac delivery and activity of caspase9 and caspase3 were increased on L02 cell membrane. In vivo, hepatic failure and obvious endotoxemia were induced by injection of more than 200 mg/kg D-GalN. Hepatic mitochondria smac was reduced with dosage of D-GalN and, on the contrary, the activity of caspase3 was increased. D-GalN at 200 mg/kg increased Caspase9 while D-GalN at 300 mg/kg decreased caspase9. Mitochondria signal channel plays an important role in the endotoxin-induced apoptosis of hepatic cells by promoting the release of smac from mitochondria to cytoplasm and activating caspase9 and caspase3 in its low-level channel.
Apoptosis/*drug effects ; Carrier Proteins/*metabolism ; Caspase 3/metabolism ; Caspase 9/metabolism ; Cell Line ; Cells, Cultured ; Endotoxins/*pharmacology ; Liver/cytology ; Liver/*metabolism ; Liver/pathology ; Liver Failure/chemically induced ; Liver Failure/pathology ; Mitochondrial Proteins/*metabolism ; Rats, Wistar