The strain Acidiphilium cryptum DX1-1 producing PHB was irradiated respectively by UV and Co60 to raise PHB production. The results indicated that the effect of UV better than using Co60. One strain of the UV mutagenized called UV60-3 has the highest PHB production yield, showing final PHB concentra- tion of 28.56 g/L, 1.45 times higher than that of original strain. FT-IR spectroscopy analysis shows that the polymers obtained from the strain DX1-1 have the same IR spectra of standard PHB. Further research about the best appropriate C/N ratio of the mutant was done. The optimum ratio of C/N was about 3.76, the final PHB concentration reaches to 30.57 g/L.

To study the proliferation characteristics of PPV in differently infected way and the variance of concentrations in different cells. A strain of porcine parvovirus(PPV) was adapted to PK-15 cells, and a Real-time fluorescent quantitative PCR (FQ-PCR) assay was developed based on the specific region of the NS1 gene of PPV to quantify the PPV. The FQ-PCR was used to measure the viral concentration of virus-infected cells by simultaneous or step by step inoculation and plot one-step growth curves. The proliferation characteristics of PPV strain in different cells lines (HeLa, MDBK, PK-15 ,ST, F81, BHK-21 and Marc-145) was also compared. The results showed the PK-15 cell -adapted strain of PPV produced CPE after 12 passages, and maintained stable CPE at the following 10 messages. The one-step growth curve showed that the virus concentration of simultaneous inoculation was higher than that of the step-by-step inoculation, and the proliferation cycle of step-by-step inoculation was shorter. The proliferation ability of PPV strain in different cells showed that CPE appeared first inPK-15, followed by ST, HeLa and MDBK, and the virus concentration was highest in ST, followed byPK-15, MDBK and HeLa. NO proliferation was observed in F81, BHK-21 and Marc-145 cells. These findings lay a material foundation for the basic researches on PPV and the development of vaccine.
Animals ; Cell Line ; Cricetinae ; Cytopathogenic Effect, Viral ; DNA, Viral ; genetics ; Female ; Haplorhini ; Humans ; Male ; Parvoviridae Infections ; virology ; Parvovirus, Porcine ; genetics ; physiology ; Real-Time Polymerase Chain Reaction ; Reproducibility of Results ; Sensitivity and Specificity ; Swine ; Viral Proteins ; genetics ; Virus Replication

To investigate the distribution of mitochondria DNA 4 977 bp deletion, a common deletion (CD), in normal populations of Chinese, human peripheral blood samples from sixty healthy donors were collected, and levels of the CD in genomic DNA from the samples were detected using real-time PCR. The results showed that the CD was found in 27 health donors, with its positive rate being 45% (27/60). The CD ratio was between 0 and 0.00308%, and not affected by age and gender in sixty healthy donors. Our studies indicate that the CD ratio is low, and do not show the age-dependent accumulation and any gender difference in peripheral whole blood from the normal Chinese population.
Base Sequence ; DNA Primers ; DNA, Mitochondrial ; genetics ; Female ; Humans ; Male ; Real-Time Polymerase Chain Reaction ; Sequence Deletion

OBJECTIVETo investigate whether p53 pathway participates in the effect of emodin on vascular smooth muscle cell proliferation.

METHODSThe effects of emodin on vascular smooth muscle cell proliferation were evaluated by cell count, senescent-associated beta-galactosidase staining, and annexin V staining. DNA synthesis was determined by (3)H-thymidine corporation, cell cycle was analyzed by FACS, the p53 protein level was measured by Western blot and cDNA expression array technology was used to demonstrate the effect of emodin on the simultaneous expression of a large number of genes in cultured vascular smooth muscle cells.

RESULTSEmodin at 1.6-3.1 microg/ml inhibited VSMC growth, at 6.3-12.5 microg/ml promoted VSMC aging and induced VSMC apoptosis at 25.0 microg/ml 24 hours after exposure. Unscheduled DNA synthesis, which was a sensitive indicator for DNA injury, was observed in VSMC following 24 hours emodin exposure. The mRNA and protein levels of p53 were up-regulated in a concentration-dependent manner. Proliferation/carcinogenesis-related genes were down-regulated and other genes related to cell senescence, apoptosis, and DNA damage/repair were up-regulated in VSMC after exposure to emodin for 24 hours. Emodin readily permeated VSMC membrane and mostly located in the cytoplasm and few of them in the nucleus.

CONCLUSIONSThe p53 pathway in VSMC was activated post emodin exposure in a concentration-dependent manner and which might be responsible for the observed antiproliferative effects of emodin in vascular smooth muscle cells.

Apoptosis ; drug effects ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; DNA Damage ; Emodin ; pharmacology ; Humans ; Myocytes, Smooth Muscle ; cytology ; drug effects ; metabolism ; RNA, Messenger ; metabolism ; Tumor Suppressor Protein p53 ; metabolism

Objective:To assess trend of clinical features,diagnosis,treatments and outcomes for in-hospital patients of acute ST-segment elevation myocardial infarction (STEMI) in Hebei province from 2001 to 2011.Methods:Our research was based on the information of China PEACE retrospective acute myocardial infarction (AMI) study.We conducted an analysis from 8 hospitals in Hebei province including 1 third class hospital and 7 second class hospitals for STEMI patients who were diagnosed,treated and discharged in those hospitals in 2001,2006 and 2011.The clinical features,process of diagnosis and treatment and outcomes were summarized.Results:A total of 832 medical records were enrolled.During 2001 to 2011,the mean age for in-hospital STEMI patients was increased as 63.5 years in 2001,65.0 years in 2006 and 66.0 years in 2011,P=0.0097;female ratio was similar as 30.1％ in 2001,30.7％ in 2006 and 30.3％ in 2011,P=0.9846;the ratio for cardiovascular risk factors were elevated as 69.9％ in 2001,87.1％ in 2006 and 87.0％ in 2011,P＜0.0010.In patients without documented contraindications,reperfusion rate was similar,P=0.8990 and primary percutaneous coronary intervention (PCI) conduction rate was similar.The following drug therapies were increased:aspirin (P＜0.0001),clopidogrel (P＜0.0001),β-blockers (P=0.0172),statins (P＜0.0001) and ACEI/ARB (P=0.0008).In 2001,2006 and 2011,the 7-day in-hospital mortality,the ratio of death and gave-up treatment were similar,P=0.5854 and P=0.3516 respectively.Conclusion:During 2001 to 2011,the onset age and the prevalence of cardiovascular risk factors were increased in STEMI patients in Hebei province;drug therapy for secondary prevention of coronary artery disease was elevated by years while the reperfusion rate was similar and 7-day mortality was similar.

OBJECTIVETo detect the incidence of the HFE gene C282Y and H63D mutations in patients with myelodysplastic syndromes (MDS) and aplastic anemia (AA), and analyze the relationship of these mutations with iron metabolism, and organs impairment from iron overload.

METHODSThe incidence of the C282Y and H63D mutations in 271 MDS, 402 AA patients and 1615 normal subjects was measured by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) combining with DNA sequencing. Iron metabolism parameters and iron overload indices were retrospectively compared between HFE gene mutation and unmutation groups in MDS and AA patients with no transfusion history.

RESULTSNo C282Y and C282Y/H63D compound mutation was detected in all the three groups. The incidence of H63D heterozygous and homozygous genotype did not significantly differ between AA cases and controls (9.7% vs 10.2%, 0.25% vs 0.24% respectively, both P > 0.05). The frequency of H63D heterozygous genotype in MDS patients was significantly lower than that in controls (4.1% vs 10.2%, P = 0.002). H63D homozygous was not found in MDS patients. In both MDS and AA patients with no RBC transfusion history, serum ferritin (SF), transferrin saturation value (TS), serum iron concentration (SI) were close to or higher than normal; and unsaturated iron-binding capacity (UIBC) value was significantly lower. There was no significant difference in SF, SI, TS values between HFE-mutation and -unmutation MDS patients. For AA patients, only the level of SI was significantly higher in HFE-mutant group than in -unmutation group [42.6 (24.6-60.4) micromol/L vs 32.0 (8.4-63.3) micromol/L, P = 0.011]. There was no significant difference in the values of liver enzyme, fasting blood sugar (FBS), abnormal electrocardiogram (ECG), peripheral blood indices between HFE-mutation and -unmutation MDS and AA groups (all P > 0.05).

CONCLUSIONThe distribution of C282Y and H63D mutations has ethnic and genetic disparity, the frequency in Chinese population is lower than that in Caucasian. It seems that MDS and AA patients are susceptible to iron overload, in the diseases itself and the mutations of HFE gene are not the major factor for iron overload in the patients.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Anemia, Aplastic ; complications ; genetics ; Case-Control Studies ; Child ; Child, Preschool ; China ; Female ; Genotype ; Hemochromatosis Protein ; Histocompatibility Antigens Class I ; genetics ; Humans ; Iron ; blood ; Iron Overload ; etiology ; genetics ; Male ; Membrane Proteins ; genetics ; Middle Aged ; Mutation ; Myelodysplastic Syndromes ; complications ; genetics ; Young Adult

The research aimed at investigating the physicochemical properties, stability and skin penetration in vitro of total alkaloids of Sophora flavescens nanoemulsion. Prepare total alkaloids of S. flavescens nanoemulsion and detect the determination of matrine and oxymatrine in the nanoemulsion using HPLC method. Transmission electron microscopy and laser particle size analyzer were utilized to detect the shape and size of the nanoemulsion respectively. And also the stability of nanoemulsion was studied under the conditions of low temperature (4 degrees C), normal temperature (25 degrees C) and high temperature (60 degrees C). Franz diffusion cell was used to research the transdermal absorption of nanoemulsion in vitro. The results found that the nanoemulsion we prepared presented appearance of rounded, uniform; its average diameter was (15.55 +/- 2.24) nm, and particle size distribution value was 0. 161; the appearance, diameter and percentage determination of total alkaloids of S. flavescens had no variations after 15 d under 4, 25, 60 degrees C respectively. The steady-state permeation rate was 4.564 1 microg x cm(-2) x h(-1), 24 h cumulative amount of penetration was 110.7 microg x cm(-2), which was 1.86 fold of 24 h cumulative amount of aqueous solution (59.41 microg x cm(-2)). All the results demonstrated total alkaloids of S. flavescens nanoemulsion had good permeability, and could provide a new preparation for its clinical application.
Alkaloids ; chemistry ; metabolism ; Animals ; Chemical Phenomena ; Drug Carriers ; chemistry ; Emulsions ; Male ; Nanostructures ; chemistry ; Rats ; Rats, Sprague-Dawley ; Skin Absorption ; Sophora ; chemistry

OBJECTIVETo prepare Kushen-Dilong nanoemulsion and nanoemuls-ion gel, and investigate its content, physical and chemical properties. Their transdermal properties in vitro were studied as well.

METHODIPM acted as oil phase, EL35 as surfactant, EtOH as cosurfactant, Pheretima aqueous solution was added dropwise to the oil phase to prepare Kushen-Dilong nanoemulsion at room temperature using magnetic stirring. HPLC was used to determine the content of matrine and oxymatrine in the nanoemulsion. Transmission electron microscopy and laser particle size analyzer was used to determine the shape and size of the nanoemulsion. NP700 was used as substrate to prepare Kushen-Dilong nanoemulsion gel. Franz diffusion cell was used for the nanoemulsion and gel transdermal characteristics in vitro.

RESULTThe Kushen-Dilong nanoemulsion was O/W nanoemulsion, its uniform particle size was 20.6 nm with roundness appearance and stable content. The steady-state permeation rate of Kushen-Dilong nanoemulsion, nanoemulsion gel, saturated aqueous solution, hydro gel were 0.1484, 0.1183, 0.0306, 0.0321 mg x cm(-2) x h(-1), respectively.

CONCLUSIONThe 24 h cumulative infiltration and infiltration rate of Kushen-Dilong nanoemulsion and nanoemulsion gel were better than the saturated aqueous solution and hydro gel, which could provide a new dosage form for Kushen-Dilong transdermal drug delivery.

Animals ; Chemistry, Pharmaceutical ; Drug Carriers ; chemistry ; Drugs, Chinese Herbal ; chemistry ; metabolism ; Emulsions ; Gels ; Nanostructures ; chemistry ; Rats ; Rats, Sprague-Dawley ; Skin ; metabolism ; Skin Absorption

BACKGROUNDThe non-operation treatment of intra-abdominal trauma guided contrast enhanced ultrasound (CEUS) is one of the hottest research topic. Gelatin/thrombin/calcium (GTC) was developed as a novel haemostatic agent for non-operable intra-abdominal trauma. We hypothesized that GTC can achieve haemostasis (without the use of pressure) within a short time in a large wound model by percutaneous injection under CEUS guidance.

METHODSForty Wister rats received large liver injuries by haemostatic clamp and were randomly divided into four groups, according to the haemostatic agent used. These included normal saline (NS) group A, lyophilising thrombin powder (LTP) group B, GTC group C, and absorbable α-cyanoacrylate (ACNA) group D. Each injury site was treated with one of the above materials and total bleeding time was recorded. All liver wounds were evaluated using CEUS at three periods: pre-injury, injury and post-treatment. The liver wounds were also evaluated by histology 3, 6, and 9 days after injury and the extents of abdominal adhesions were recorded.

RESULTSThe sensitivity of CEUS (100%) in detecting blunt traumatic liver lesions was significantly higher than conventional ultrasound (42.5%). Bleeding times at the injury site in the GTC group C ((129.3 ± 14.0) seconds) and ACNA group D ((5.2 ± 1.0) seconds) were significantly shorter than those in the NS group A ((369.5 ± 48.8) seconds, P < 0.01) and LTP group B ((324.7 ± 52.22) seconds, P < 0.01). The LTP group B showed no significant difference compared with the NS group A. Gross examination of liver tissue revealed that there were fewer intra-abdominal adhesions in the GTC group C (10%) than in the ACNA group D (100%). Histopathologic examination showed that GTC was completely absorbed after nine days.

CONCLUSIONSGTC, delivered by percutaneous injection under CEUS, may achieve haemostasis (without the use of pressure) within a short time in a large wound model. GTC is absorbable and may prevent intra-abdominal adhesions. Therefore, it may be the optimal choice for first aid treatment of large abdominal wounds in the setting of blunt trauma.

Animals ; Calcium ; administration & dosage ; therapeutic use ; Gelatin ; administration & dosage ; therapeutic use ; Hemorrhage ; diagnostic imaging ; drug therapy ; Hemostatics ; administration & dosage ; therapeutic use ; Injections ; Liver ; diagnostic imaging ; injuries ; Male ; Random Allocation ; Rats ; Rats, Wistar ; Thrombin ; administration & dosage ; therapeutic use ; Ultrasonography

OBJECTIVETo analyze the expression of genes from chromosomal region 22q11.2 and assess the association between mutation(s) of particular gene(s) from this region and malformations of the urinary system.

METHODSExpression of rat homologs of 33 genes from above region was determined in kidney tissues derived from rats of different fetal development ages (E13, E15, E19) and adulthood with reverse transcriptase-PCR. Potential mutation(s) in candidate gene SNAP29, whose expression pattern appeared to be unique, was screened in 44 patients and 220 normal controls with PCR-single strand conformation polymorphism (SSCP). Suspected positive regions were sequenced to verify the mutations.

RESULTSNine genes showed no expression throughout the whole development process; 18 genes with various expression levels showed continuous expression from the beginning of development; 6 genes only expressed for a short time, among which SNAP29 was selected for mutation screening. Upon sequencing, three mutations were identified from the 44 patients, including a G to A transition (GAG to AAG) in exon 2, and two A to G transitions (AGC to GGC) in exon 3.

CONCLUSIONThrough systematic analysis of the expression of genes from chromosomal region 22q11.2, the SNAP29 gene was found to have a potential role in the development of genitourinary system. Two missense mutations were identified in three patients. These included one in exon 2 (featuring cryptorchidism), and the other in exon 3 (featuring cryptorchidism and hypospadia). Neither of the mutations was found in the normal controls. The results suggested that mutation(s) of gene(s) from chromosomal region 22q11.2 may play an important role in the genesis of genitourinary malformations.

Animals ; DNA Mutational Analysis ; methods ; Exons ; genetics ; Female ; Humans ; Male ; Membrane Glycoproteins ; Membrane Proteins ; genetics ; Mice ; Platelet Glycoprotein GPIb-IX Complex ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Polymorphism, Single-Stranded Conformational ; Qb-SNARE Proteins ; genetics ; Qc-SNARE Proteins ; genetics ; Urogenital Abnormalities ; genetics