To develop a parent-metabolite pharmacokinetic model for risperidone (RIP) and its major active metabolite (9-hydroxyrisperidone) and investigate their pharmacokinetics characteristics in healthy male volunteers, twenty-two healthy volunteers were orally given a single dose of 2 mg RIP. Plasma samples were collected in the period of 96 hours and concentrations of RIP and 9-hydroxyrisperidone were measured by a validated HPLC/MS method. CYP2D6 phenotypes were identified by the T1/2 of RIP and 9-hydroxyrisperidone according to the literature. Model structure identifiability analysis was performed by the similarity transformation approach to investigate whether the unknown parameters of the proposed model could be estimated from the designed experiment. Pharmacokinetics parameters were estimated using weighted least squares method, and the final pharmacokinetics model were tested and evaluated by Monte Carlo simulation. Eighteen volunteers were phenotyped as extensive metabolizers (EM) and four volunteers were identified as intermediate metabolizers (IM). The final model included central and peripheral compartment for both parent (RIP) and metabolite (9-hydroxyrisperidone) respectively. Model structure identifiability analysis indicated that the proposed model was local identifiable. However, if the ratio of RIP converted to 9-hydroxyrisperidone was assumed to be 32% in EM, and 22% in IM, the model could be globally identifiable. The predicted time-concentration curve and AUC(0-t), C(max), T(max) of RIP and 9-hydroxyrisperidone estimated by the established model were in agreement with the observations and noncompartment analysis. Rate constant of RIP conversion to 9-hydroxyrisperidone was (0.12 +/- 0.08) h(-1) and (0.014 +/- 0.007) h(-1) for EM and IM, respectively. Elimination rate constants of RIP were (0.25 +/- 0.18) and (0.05 +/- 0.23) h(-1) for EM and IM, respectively. Model validation result showed that all parameters derived from the concentration data fitted well with the theoretical value, with mean prediction error of most PK parameter within +/- 15%. The established model well defined the disposition of RIP and 9-hydroxyrisperidone simultaneously and showed large inter-individual pharmacokinetics variation in different CYP2D6 phenotype. The model also provide a useful approach to characterize pharmacokinetics of other parent-metabolite drugs.
Adult ; Cytochrome P-450 CYP2D6 ; physiology ; Humans ; Isoxazoles ; pharmacokinetics ; Male ; Models, Biological ; Monte Carlo Method ; Paliperidone Palmitate ; Pyrimidines ; pharmacokinetics ; Risperidone ; pharmacokinetics

OBJECTIVETo seek a new method for the categorization of burn severity.

METHODSBurn patients hospitalized in our center from December of 1958 to December of 2004 were enrolled in the study, and they were divided into different age groups according to same mortality, then the patients in each group were subdivided into 4 groups according to the burn severity: i.e., mild burns, moderate burns, severe burns, serious severe burns. The total burn area, the number of cases, the mortality, and the area of DI degree burns were statistically analyzed in each subgroup, and the scope in total burn area and area of III degree burns were taken as standards to define the degree of burns. The logistic regression equation was established with probability of death as the variable, and age, total burn area, burn area of different degrees as concomitant variables to form a logistic regression formula. It was used to predict the probability of death of patients hospitalized in 2005, 50 as to check whether the corresponding indices of these patients were consistant with above standard of categorization into degrees, and to judge hum severity of the patients who had concomitant inhalation injury, severe associated injury, or those with serious disease before burns.

RESULTSThe patients were divided into three groups: less than 2 years of age (including 2 years of age), 2 to 55 years of age(including 55 years of age), and older than 55 years of age groups. The classification standard of burn area was shown in table 2 of the article. The probability of death and corresponding indices predicted hy the logistic regression equation were highly coincident with our standard. Patients with moderate inhalation injury could be regarded as patients with severe or most severe burns, while severity of those with mild inhalation injury could be determined by burn area alone.

CONCLUSIONThe logistic regression equation is a good method to predict the severity of burn patients, with reasonable age specificity grouping, and accurate and practical scoring of division for corresponding burn severity.

Adolescent ; Adult ; Burns ; classification ; Child ; Child, Preschool ; Humans ; Injury Severity Score ; Logistic Models ; Middle Aged ; Young Adult

The gene encoding a extremely thermostable and acid-stable alpha-Amylase was amplified by PCR using hyperthermophilic archaebacterium pyrococcus furiosus genomic DNA as template. Then the gene was cloned into the vector of pPIC9K. The recombinant vector pPIC9K-amy was then transformed into E. coli DH5alpha strain. Sequencing test showed that the a-amylase gene cloned consisted of 1305 base pairs and the mature protein encoded by the gene consisted of 435 amino acids. The recombinant vector was transformed into chromosome of methylotrophic yeast Pichia pastoris GS115 strain. Regulated by the alpha-Factor, promoter of AOX1 gene and termination signal of yeast genomic, the recombinant a-Amylase was expressed and excreted out of the cells. The expression of the recombinant alpha-amylase was strictly induced by methanol. As induction time increased, the activity of amylase per milliliter medium went up accordingly. After 7 days induction, the activity of the amylase reached the max. The recombinant alpha-amylase exhibited maximal activity at 90 to approximately 100 degrees C and at pHranging from 4.5 to 5.0. The enzyme is so thermostable that after disposed at 100 degrees C for 5 hours over 60% of activity was retained.
Bacterial Proteins ; genetics ; metabolism ; Cloning, Molecular ; Enzyme Stability ; Genetic Vectors ; Hot Temperature ; Hydrogen-Ion Concentration ; Pichia ; genetics ; metabolism ; Polymerase Chain Reaction ; Pyrococcus furiosus ; enzymology ; genetics ; Recombinant Proteins ; genetics ; metabolism ; alpha-Amylases ; genetics ; metabolism

AIMTo develop a pharmacokinetic model for the enterohepatic circulation of mycophenolic acid (MPA).

METHODSTwenty healthy volunteers were orally given a single dose of 500 mg mycophenolate mofetil. Plasma samples were collected during 48 hours and MPA concentration was measured by HPLC method. Pharmacokinetic (PK) model was established based on physiological and biopharmaceutical consideration and PK parameters were obtained using nonlinear mixed effect model.

RESULTSThe proposed model included an intestinal compartment and gall bladder compartment in addition to the central compartment. The predicted time-concentration curve and AUC0-t, Cmax, Tmax estimated by the established model were in agreement with the observations.

CONCLUSIONThe established model was well defined for the MPA disposition and could afford a useful approach for the further clinical investigation.

Adult ; Area Under Curve ; Enterohepatic Circulation ; physiology ; Glucuronides ; pharmacokinetics ; Humans ; Immunosuppressive Agents ; pharmacokinetics ; Male ; Models, Biological ; Mycophenolic Acid ; administration & dosage ; analogs & derivatives ; blood ; pharmacokinetics

OBJECTIVETo elevate the immunological effect of subunit influenza vaccine in infants and aged people (over 60) using liposomal adjuvant in the context of its relatively low immunity and to investigate the relation between vaccine antigens and liposomal characteristics.

METHODSSeveral formulations of liposomal subunit influenza vaccine were prepared. Their relevant characteristics were investigated to optimize the preparation method. Antisera obtained from immunizinged mice were used to evaluate the antibody titers of various samples by HI and ELISA.

RESULTSLiposomal trivalent influenza vaccine prepared by film evaporation in combinedation with freeze-drying significantly increased its immunological effect in SPF Balb/c mice. Liposomal vaccine stimulated the antibody titer of H3N2, H1N1, and B much stronger than conventional influenza vaccine. As a result, liposomal vaccine (mean size: 4.5-5.5 microm, entrapment efficiency: 30%-40%) significantly increased the immunological effect of subunit influenza vaccine.

CONCLUSIONThe immune effect of liposomal vaccine depends on different antigens, and enhanced immunity is not positively correlated with the mean size of liposome or its entrapped efficiency.

Animals ; Influenza A Virus, H1N1 Subtype ; immunology ; Influenza A Virus, H3N2 Subtype ; immunology ; Influenza B virus ; immunology ; Influenza Vaccines ; administration & dosage ; immunology ; Liposomes ; Mice ; Mice, Inbred BALB C ; Orthomyxoviridae Infections ; prevention & control ; Specific Pathogen-Free Organisms ; Vaccines, Subunit ; administration & dosage ; immunology

BACKGROUNDTrans-arachidonic acids (TAAs), newly discovered markers of nitrative stress and the major products of nitrogen dioxide (NO2(·))-mediated isomerization of arachidonic acid (AA), represent a new mechanism of NO2(·)-induced toxicity. It has been reported that TAAs were generated in oxygen-induced microvascular degeneration model and TAAs were also generated in a diabetic retinopathy (DR) model. In this study, we examined high glucose-induced nitrative stress damage and TAAs levels and explored the possible mechanisms for DR caused by reactive nitrogen species.

METHODSDiabetic rats were induced by intraperitoneal injection of streptozotocin (STZ) at 60 mg/kg. Bovine retinal capillary endothelial cells (BRECs) were selectively cultured and incubated with normal or high glucose. The serum TAAs and AA in diabetic rats were measured by the gas chromatography and mass spectrometry (GC/MS) method. The ratio of peak area of TAAs to AA with selected ion of 79 was estimated by a group t-test. Thrombospondin-1 (TSP-1) in the rat retinas and BRECs extracts were examined by Western blotting. The phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2) protein was examined by Western blotting in BRECs incubated with high glucose.

RESULTSThe TAAs to AA ratio (TAAs/AA) was significantly increased in the serum at 8, 12 and 16 weeks after STZ injection (P < 0.05), with no noticeable change found at 2 or 4 weeks (P > 0.05). Expression of TSP-1 in the retina of diabetic rats was progressively elevated according to the duration of diabetes. TSP-1 expression was increased in BRECs incubated with high glucose at 48 hours. Moreover, high glucose also increased ERK1/2 expression, which peaked at 30 minutes and then decreased in the following 48 hours.

CONCLUSIONAn elevation of TAAs/AA is associated with high glucose-induced nitrative stress, which probably involves upregulation of TSP-1 through activating ERK1/2.

Animals ; Arachidonic Acid ; metabolism ; Blotting, Western ; Cattle ; Cells, Cultured ; Diabetes Mellitus, Experimental ; metabolism ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Gas Chromatography-Mass Spectrometry ; Male ; Rats ; Rats, Sprague-Dawley ; Reactive Nitrogen Species ; metabolism ; Streptozocin ; Thrombospondin 1 ; genetics ; Up-Regulation

OBJECTIVETo explore F (13) A gene mutation in a pedigree with hereditary coagulation factor XIII (FXIII) deficiency.

METHODSThe FXIII deficiency was diagnosed by clot solubility test and other standard laboratory clotting tests. All exons, exon-intron boundary sequences of F(13) A gene were amplified by PCR and the products were sequenced directly. Any mutation identified by direct sequencing was confirmed by reverse sequencing. The mutation identified in the proband was screened in the family members.

RESULTSThe assays of PT, Qiulan, fibrinogen leveling, platelet counts, bleeding time were normal and the clot solubility test was positive in the proband. The homozygous deletion of 33 nucleotides (127067de133) in exon 10 of F(13) A gene which resulted in deletion of 11 amino acids in FXIIII A protein with 720aa residues was identified in the proband. Family studies showed that the mutation was inherited from the parents both of whom carried the heterozygous deletion mutation.

CONCLUSIONThe homozygous 127067de133 mutation of F(13) A gene is responsible for the disorder of the pedigree.

Adolescent ; Factor XIII ; genetics ; Factor XIII Deficiency ; genetics ; Heterozygote ; Homozygote ; Humans ; Male ; Pedigree ; Sequence Deletion

OBJECTIVETo investigate whether the polymorphism of DNA repair genes XPC (Ala499Val and Lys939Gln) and XPG (His1104Asp) is associated with the susceptibility to hepatocellular carcinoma (HCC).

METHODSA hospital-based case-control study was conducted in 500 cases with HCC and 507 controls. Genotypes of XPC and XPG were determined by real-time polymerase chain reaction with the TaqMan MGB probe.

RESULTSCompared to the CC genotype, the CT genotype and the TT genotype of XPC Ala499Val were not associated with the susceptibility to HCC (adjusted OR = 1.34, 95% CI: 0.85-2.12; adjusted OR = 1.30, 95% CI: 0.68-2.51, respectively). Compared to the AA genotype, the AC genotype and the CC genotype of Lys939Gln were not associated with the susceptibility to HCC (adjusted OR = 1.20, 95% CI: 0.78-1.85; adjusted OR = 1.81, 95% CI: 0.88-3.73, respectively). Compared to the CC genotype, the CG genotype and the GG genotype of XPG His1104Asp were not associated with the susceptibility to HCC (adjusted OR = 0.85, 95% CI: 0.56-1.27; adjusted OR = 1.12, 95% CI: 0.67-1.87, respectively) However, the stratified analysis revealed that the females with the AC+CC genotype of XPC Lys939Gln had increased risk of HCC compared to those with AA genotype (OR = 2.17, 95% CI: 1.01-4.64).

CONCLUSIONOur results suggest that XPC and XPG polymorphisms do not independently affect on the susceptibility to HCC, but the joint effect of C allele of XPC Lys939Gln and female may modify the risk of HCC.

Carcinoma, Hepatocellular ; genetics ; Case-Control Studies ; DNA Repair ; DNA-Binding Proteins ; genetics ; Endonucleases ; genetics ; Female ; Genetic Predisposition to Disease ; Genotype ; Humans ; Liver Neoplasms ; genetics ; Male ; Middle Aged ; Nuclear Proteins ; genetics ; Polymorphism, Single Nucleotide ; Transcription Factors ; genetics

Objective: Children with autism spectrum disorder(ASD) who participated in the " Play-based Communication and Behavior Intervention(PCBI)" were studied in order to evaluate the impact of caregiver factors on the efficacy of the PCBI very early intervention model systematically. Methods: Seventy-six children with ASD aged 8-30 months were recruited in this study at Brain Hospital Affiliated to Nanjing Medical University form December 2017 to June 2018, and they were trained for PCBI very early intervention.The Autism Treatment Evaluation Checklist (ATEC) and the Gesell Developmental Schedules were used to evaluate the efficacy of the PCBI very early intervention.Parenting Stress Index Short Form, General Self-Efficacy Scale and the self-developed Caregiver Training Course Evaluation Scale and the Homework Completion Level Scale were used to assess the level of parental stress, self-efficacy, curriculum satisfaction and acceptance.A pair of sample t-test was used to analyze the ASD symptoms, developmental level and caregiver factors of ASD children before and after intervention.Multivariate regression analysis was used to analyze the caregiver factors and the effects of PCBI very early intervention. Results: (1) At the end of 12 weeks of PCBI very early intervention, the score of the ATEC (17.36±15.98) was lower than that before intervention (78.43±22.84), and the difference was statistically significant(P<0.05); the adaptive developmental quotient(5.90±15.45), the fine action developmental quotient(5.13±19.89), the language developmental quotient (12.43±20.60) and the personal social developmental quotient(4.79±16.02) scores of the Gesell Developmental Schedules were higher than those before intervention (73.08±20.54, 77.33±23.63, 52.24±19.12, 71.79±20.81), and the differences were statistically significant (all P<0.05). (2) At the end of PCBI very early intervention, there was a significant decrease in the scores of Parenting Stress Index Short Form, and the differences were statistically significant (4.84±14.69 vs.94.05±29.67, P<0.05). (3) Acceptance of the intervention model, and mastery of the skills in the caregiver factors had a positive predictive effect on the effect of PCBI very early intervention (t=3.068, 10.468, all P<0.05). Conclusions PCBI very early intervention can effectively reduce the parenting pressure of the caregivers, and in the PCBI very early intervention, the better the mastery of skills and the completion of homework, the better the efficacy of intervention.

OBJECTIVE: To study the value of airway pH monitoring in determining the association between chronic cough and laryngopharyngeal reflux (LPR) in children. METHODS: A total of 274 children with chronic cough who were treated from January 2016 to December 2019 were enrolled. The DX-pH detection system was used to conduct 24-hour airway pH monitoring. The association between chronic cough and LPR was analyzed. RESULTS: Among the 274 children, there were 168 boys and 106 girls, with a median age of 62.8 months and a median airway pH value of 7.3. Of all the 274 children, 99 (36.1%) had LPR, and the incidence rate of LPR was 36.9% (62/168) in boys and 34.9% (37/106) in girls ( CONCLUSIONS LPR is highly associated with the development of chronic cough, and airway pH monitoring may be a safe and effective method for the diagnosis of LPR.
Child ; Child, Preschool ; Chronic Disease ; Cough/etiology* ; Esophageal pH Monitoring ; Female ; Humans ; Hydrogen-Ion Concentration ; Laryngopharyngeal Reflux ; Male