Objective To investigate the in vitro pharmaceutical properties(protein content and purity,biological activity)and in vivo pharmacodynamics(procoagulant activity)of recombinant human thrombin.Methods The content of protein in recombinant human thrombin was determined by folin-reagent method.Sodium dodecyl sulphate-polyacrylamide gel electrophoresis was used to determine its purity.Human thrombin activity test was applied to detect its biological activity.Eighteen male New Zealand rabbits were randomly divided according to body mass into blank group(0.9％NaCl solution),experimental-L group(200 U·mL-1 recombinant human thrombin)and experimental-H group(1 000 U·mL-1 recombinant human thrombin),6 rabbits in each group.The procoagulant effects of recombinant human thrombin were investigated in the rabbit liver injury model.Results The protein content of recombinant human thrombin was(1.980±0.024)mg per container,its biological activity was(5764.3±197.7)U per container and its biological specific activity was(2 911.2±99.8)U·mg-1.The target protein(35 kD in non-reductive electrophoresis)has a single band and few miscellaneous proteins were found on the gel.Compared with the blank group,the mean hemostasis time and mean blood loss weight of rabbit liver wound were reduced by 57.0％and 87.7％in the experimental-L group,which in the experimental-H group were reduced by 71.8％and 91.9％.Conclusion Recombinant human thrombin has high protein purity and specific biological activity,and has significant procoagulant activity in vivo.

Objective To investigate the in vitro pharmaceutical properties(protein content and purity,biological activity)and in vivo pharmacodynamics(procoagulant activity)of recombinant human thrombin.Methods The content of protein in recombinant human thrombin was determined by folin-reagent method.Sodium dodecyl sulphate-polyacrylamide gel electrophoresis was used to determine its purity.Human thrombin activity test was applied to detect its biological activity.Eighteen male New Zealand rabbits were randomly divided according to body mass into blank group(0.9％NaCl solution),experimental-L group(200 U·mL-1 recombinant human thrombin)and experimental-H group(1 000 U·mL-1 recombinant human thrombin),6 rabbits in each group.The procoagulant effects of recombinant human thrombin were investigated in the rabbit liver injury model.Results The protein content of recombinant human thrombin was(1.980±0.024)mg per container,its biological activity was(5764.3±197.7)U per container and its biological specific activity was(2 911.2±99.8)U·mg-1.The target protein(35 kD in non-reductive electrophoresis)has a single band and few miscellaneous proteins were found on the gel.Compared with the blank group,the mean hemostasis time and mean blood loss weight of rabbit liver wound were reduced by 57.0％and 87.7％in the experimental-L group,which in the experimental-H group were reduced by 71.8％and 91.9％.Conclusion Recombinant human thrombin has high protein purity and specific biological activity,and has significant procoagulant activity in vivo.

There is no doubt that the traditional Chinese medicine(TCM) is effective, practical and scientific after it was used for thousands of years. However, the mechanisms of action of many TCM are still unclear because of their multi-component, multi-target and multi-level features, which hinder the modernization and internationalization of the TCM. Proteomics is to analyze the composition and activity of intracellular proteins which are changing dynamically from a holistic perspective. It is consistent with the holistic and dynamic views of the TCM and brings about the hope of clarifying the mechanism of action of the TCM. In recent years, great progress has been made in the application of proteomics to determine the mechanism of the TCM. This article introduced the core technologies of proteomics and systematically summarized the applications of proteomics in the study of the mechanism of the Chinese medicinal formulae, single Chinese medicine and monomeric compounds from the TCM to provide innovative ideas and methods for reference.

OBJECTIVETo investigate the species and percentage changes of pathogens in blood cultures from the pediatric hematology ward, and to analyze the drug resistance of main pathogens and the risk factors for positive blood culture (sepsis).

METHODSA retrospective analysis was performed to analyze the species and drug sensitivity of the pathogens isolated from 2358 blood cultures from the pediatric hematology ward of the West China Second University Hospital between 2008 and 2011, as well as the related clinical data.

RESULTSA total of 110 strains of pathogens were isolated, with Escherichia coli (16 strains), Pseudomonas aeruginosa (12 strains) and Staphylococcus epidermidis (8 strains) being the most common ones. From 2008 to 2011, the percentage of Gram-positive bacteria decreased, while the percentage of Gram-negative bacteria increased. The detection rates of methicillin-resistant coagulase-negative Staphylococci and methicillin-resistant Staphylococcus aureus were 69% and 43% respectively, but both were sensitive to vancomycin. The detection rates of extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli and ESBL-producing Klebsiella pneumoniae were 69% and 62% respectively, but both were sensitive to imipenem and meropenem. Malignant tumor was a risk factor for positive blood culture (OR=3.564, P<0.05).

CONCLUSIONSA wide range of pathogens are responsible for bloodstream infection in the pediatric hematology ward and the percentages of bacteria are changing; these pathogens have a high drug resistance rate. Malignant tumor is a risk factor for positive blood culture in the pediatric hematology ward.

Adolescent ; Bacteremia ; etiology ; microbiology ; Bacteria ; drug effects ; isolation & purification ; Child ; Child, Preschool ; Drug Resistance, Bacterial ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Retrospective Studies

OBJECTIVETo investigate the change in dendritic cells (DCs) in children with chronic immune thrombocytopenia (cITP) and the effect of glucocorticoid on DCs in children with cITP.

METHODSFifteen children with cITP and 20 healthy controls were included in the study. Flow cytometry was used to measure the DC subsets count in the 15 children with cITP before and after glucocorticoid treatment as well as the corresponding values in the 20 healthy controls. The DCs derived from peripheral blood monocytes in children with cITP were cultured in vitro and collected, and their immunophenotypes were determined by flow cytometry.

RESULTSBefore glucocorticoid treatment, the children with cITP showed no notable change in the absolute count of myeloid DCs (mDCs) but showed decreased absolute count of plasmacytoid DCs (pDCs) and increased mDC/pDC ratio compared with the healthy controls (P<0.05). After glucocorticoid treatment, the children with cITP demonstrated increased absolute count of pDCs and decreased absolute count of mDCs and mDC/pDC ratio compared with before treatment (P<0.05). Before glucocorticoid treatment, the children with cITP had significantly higher positive rates of HLA-DR, CD80, CD83 and CD86 on peripheral blood DCs than the healthy controls (P<0.01). All the positive rates were significantly decreased after glucocorticoid treatment (P<0.01), so that there was no significant difference from the healthy controls (P>0.05).

CONCLUSIONSDisproportion and functional disturbance of DC subsets is associated with the pathogenesis of cITP in children. Glucocorticoid can strengthen the immunosuppression of DCs in children with cITP, which may contribute to the effectiveness of glucocorticoid as a treatment.

Adolescent ; Child ; Child, Preschool ; Chronic Disease ; Dendritic Cells ; drug effects ; immunology ; Female ; Glucocorticoids ; pharmacology ; Humans ; Immunophenotyping ; Male ; Thrombocytopenia ; drug therapy ; immunology

OBJECTIVETo study the clinical characteristics and pathogens of invasive fungal infection in children.

METHODSThe clinical data of 104 children who suffered from invasive fungal infections between 2008 and 2012 was retrospectively reviewed.

RESULTSOf the 104 cases, 20 occurred in neonates, 48 in infants and 36 in preschool and school-aged children (old-aged children). Prematurity (70%), hyaline membrane disease (45%) and pneumonia (30%) were commonly comorbid in the neonate group. In addition, the percentage of cases receiving total parenteral nutrition was higher in the neonate group than in the other two age groups (P<0.01). Mechanical ventilation was more frequent in neonate and infant groups than in the old-aged children (P<0.01). Hematological malignancy was the most common underlying disease, and the percentage of children who had neutropenia and accepted chemotherapy was higher in the old-aged children than in the other two age groups (P<0.05). Lung infection was the most common (61.5%), followed by sepsis (14.4%) and intestinal tract infection (12.5%), while nervous system infections were found only in old-aged children. A total of 105 strains of fungi were isolated from the 104 patients, including Candida (n=90, 85.7%), Cryptococcus (n=6) and others (n=9). The most commonly isolated species was Candida albicans (n=52, 49.5%). Non-Candida albicans Candida accounted for 36.2% (n=38). The rate of susceptibility of Candida species to 5-fluorocytosine and amphotericin B was higher than fluconazole.

CONCLUSIONSInvasive fungal infections can occur in children at various ages. There are differences in the risk factors for invasive fungal infections between age groups. Candida species are the main pathogens of childhood invasive fungal infections, and both Candida albicans and non-Candida albicans Candida are common. Fluorocytosine and amphotericin B are sensitive antifungal agents for infections caused by Candida species.

Adolescent ; Child ; Child, Preschool ; Female ; Fungi ; drug effects ; isolation & purification ; Humans ; Infant ; Infant, Newborn ; Male ; Microbial Sensitivity Tests ; Mycoses ; drug therapy ; etiology ; microbiology ; Prognosis ; Risk Factors

Objective To dynamically analyze the evolutionary process of cerebral edema absorption and the level of local iron in rats with intra-cerebral hematoma by high-field strength 7 Tesla MRI and explore the characteristics and mechanism of secondary injury after intra-cerebral hematoma.Methods Sixteen adult SD rats (about 150 g) were randomly divided into experimental group (n=10) and control group (n=6).Rat models in the experimental group were established by performing injection of 50 μL their own venous blood into their right caudate nucleus accurately. Rats in the control group were used normal saline,instead.After that,head MRI (T2 and T2-star scans) was performed 1,2,3,7 and 14 d after the injection; their imaging features were compared. Results Nine rats in the experimental group survived and 1 died after the operation; in the early days (within 3 d), the T2 weighing imaging showed that the time of relaxation surrounding the hematoma was longer than that in control group,suggesting that the zone of the edema surrounding the hematoma became more clearly.In the early days (within 3 d),T2-weighted imaging was clear,and the time of relaxation surrounding the hematoma increased rapidly,steadily improved 3 d after the operation and reached its peak level 7 dafter the operation; the damage area absorption decreased steadily but turned widening 3 d later and reached the peak 7 d later.T2-star value reached the peak rapidly 3 d after the operation,and then,moderated the downturn.The rats in the control group showed no obvious signal changes under MRI,except those with needle tract injury. Conclusion Secondary injury after intra-cerebral hemorrhage shows a rapidly injury progress in the short terrn at first,and then,has intensify again after a stable period; the local iron diffusion trend is synchronized to the secondary injury,suggesting that iron may play a key role in the mechanism of secondary brain edema.

Objective Coronary arterial plaque rupture and secondary thrombosis are the major pathogenesis of acute coronary syndrome ( ACS) . Metalloprotease ( MMPs) secreted by monocyte/ macrophage was the main predisposing factor of the plaque rupture and peroxisotne proliferator-activated receptor-γ (PPAR-γ) is involved in a variety of inflammatory cytokine gene transcriptional regulations. We explored the possible role of PPAR-γ in the regulation of MMP-9 and TIMP-1 expressed by peripheral monocyte-derived macrophages (MDMs) from patients with ACS. Methods Peripheral blood mononuclear cells were isolated from 48 patients with ACS and 28 healthy controls and stimulated by macrophage colony-stimulating factor (0. 1 μg/ml for 24 hours) to form MDMs. MDMs were then incubated under various concentrations of rosiglitazone (0, 1, 10, 20 μmol/L) for 48 hours. The concentrations of MMP-9 and TIMP-1 in the supernatant were measured by enzyme linked immunosorbent assay, and the mRNA expression of PPAR-γ, MMP-9 by RT-PCR and nuclear factor-KB P65 ( NF-kB P65 ) expression by immunohistochemistry. Results PPAR-γ mRNA expression was significantly lower while NF-kB P65 and MMP-9 expression as well as MMP-9 and TIMP-1 concentrations in supernatant were significantly higher in ACS group than those in control group (all P <0. 05). After rosiglitazone intervention, PPAR-γ mRNA expression was significantly upregulated in both ACS and control groups in a dose-dependent manner. Both the MMP-9 concentration in the supernatant and MMP-9 mRNA expression were reduced post intervention with rosiglitazone in both groups. The TIMP-1 mRNA expression and concentration in supernatant were not affected by rosiglitazone in both groups. Rosiglitazone induced significant downregulation of NF-kB P65 expression in both groups. Conclusion Rosiglitazone intervention may downregulate MMP-9 expression by upregulating PPAR-γ expression, and by downregulaiting NF-kB expression in MDMs isolated from patients with ACS.

Objective To discuss the valuable influence of the total-body massage on the facilitation of the rehabilitation for the women in puerperium.Methods Both Chinese traditional medical total-body massage and the biological information theories and its methodology were employed in dealing with 50 cases of puerperants without contraindications,who had labored naturally 12 hours before and who had received cesarean sections 48 hours before. The results were compared with the control group in which conventional preventive health care for puerperants was adopted.Results Notable differences exist in the aspect of lochia,defecation,lactation and emotions between the two groups.Conclusion Significant benefits can be obtained from the massage to facilitate the rehabilitation of the women in puerperium while promoting their quality of lives and broadening the coverage of services of the preventive health care for the puerperants.