ADP Ribose Transferases ; chemistry ; pharmacology ; Animals ; Antineoplastic Agents ; pharmacology ; Bacterial Toxins ; chemistry ; pharmacology ; Exotoxins ; chemistry ; pharmacology ; Female ; HMGN2 Protein ; chemistry ; pharmacology ; HeLa Cells ; Humans ; Immunotoxins ; pharmacology ; Mice ; Mice, Inbred BALB C ; Protein Structure, Tertiary ; Recombinant Fusion Proteins ; biosynthesis ; pharmacology ; Virulence Factors ; chemistry ; pharmacology

AIMTo explore the effect of Triptolide on airway remodeling and the expression of Phosphoinositide 3-Kinases in asthmatic rats.

METHODS40 rats were randomly divided into 5 groups (n = 8): (1) Control group; (2) Asthmatic 4 weeks group; (3) Asthmatic 6 weeks group; (4) Therapeutic 4 weeks group; (5) Therapeutic 6 weeks group. The airway resistance and eosinophilic inflammation of airway wall were observed. The airway wall thickness (WA/Pi), the bronchial smooth muscle thickness (smooth muscle area/Pi) and the number of bronchial smooth muscle nucleus (N/Pi) were measured by image analysis system. The expression of PI3K protein and mRNA were determined by immunohistochemical staining and reverse transcription-polymerase chain reaction (RT-PCR).

RESULTS(1) The expression of PI3K p85alpha protein and mRNA in asthmatic 4 weeks group and asthmatic 6 weeks group were significantly higher than control group, respectively (P < 0.01). The above-mentioned parameters of therapeutic 6 weeks group were significantly lower than those of asthmatic 4 weeks group, asthmatic 6 weeks group and therapeutic 4 weeks group, respectively (P < 0.01, P < 0.01 P < 0.05). (2) The WA/Pi, the smooth muscle area/Pi and the N/Pi of asthmatic 4 weeks group and asthmatic 6 weeks group were significantly higher than control group, respectively (P < 0.01). The above-mentioned parameters of therapeutic 6 weeks group were significantly lower than those of asthmatic 4 weeks group, asthmatic 6 weeks group and therapeutic 4 weeks group, respectively (P < 0.01). (3) The airway resistance of asthmatic 4 weeks group and asthmatic 6 weeks group were significantly higher than the control group, respectively (P < 0.01). The above-mentioned parameters of therapeutic 6 weeks group were significantly lower than those of asthmatic 4 weeks group, asthmatic 6 weeks group and therapeutic 4 weeks group, respectively (P < 0.01, P < 0.01, P < 0.05).

CONCLUSIONThe proliferation of airway smooth muscle is a remarkable character of airway remodeling in asthma. The PI3K signal pathway may be involved in the process. Triptolide may reduce AHR and decrease the proliferation of ASMCs by inhibiting the expression of PI3K. It may have potential therapeutic effects in the asthmatic airway remodeling.

Airway Remodeling ; Animals ; Asthma ; metabolism ; physiopathology ; Diterpenes ; pharmacology ; Epoxy Compounds ; pharmacology ; Male ; Phenanthrenes ; pharmacology ; Phosphatidylinositol 3-Kinase ; metabolism ; Rats ; Rats, Sprague-Dawley ; Signal Transduction

Objective To investigate the clinical features of aortic dissection (AD) and emergency treatments. Methods Data from 784 patients with aortic dissection were collected in the Department of Emergency from January 2000 through December 2009. A retrospective analysis was carried out to determine the survival rate, mortality rate and treatment efficiency. Results Pain was the most common onset symptom (77.7% , 609/784). The majority of patients (86.5%) had essential hypertension (678/784). All the patients with preoperative diagnosis of aortic dissection underwent emergency medical intervention by internists resulting in 81.5% survival rate (639/784) and 18.5% mortality rate (145/784). There were 157 patients without improvement (20.0% ) and the total efficiency rate was (83. 1% ). The efficiency rate of conventional treatment was 76.4% , while the efficiency rate of triple four-procedure treatment was 89. 8% (P<0.05). Of them, 139 patients (17. 7% ) died in the hospital. Among them,. 26 patients died within 24 hours (18.4% ) and 47 cases died within 48 hours (33. 8% ) and 66 patients died within 72 hours (47.2% ). There were 92 patients who refused treatments after diagnosis, and among them, 81 patients died within 72 hours (88.04% ). The difference in mortality rate between two groups was significant (P<0.05). Conclusions The diagnosis of aortic dissection depends on detailed history, physical examination and CT or MRI imaging. Analgesia, sedation and control of blood pressure are essential for emergency treatments. Early diagnosis and effective emergency treatments are the critical strategy for the early surgical intervention and time window for further treatment to improve the survival rate of AD.

To purify recombinant human nucleoside diphosphate kinase A (rhNDPK-A) and determine its physical and chemical characters, recombinant NDPK-A producing E. coli was cultured in 80L fermentor under high cell density culture (HCDC) conditions. The harvested cells were treated with high pressure to break the cell up, tangential-flow microfiltration to remove the bacteria debris and ultrafiltration to concentrate the filtered solution containing target protein. The crude NDPK-A was purified by ion exchange chromatography with DEAE Sepharose Fast Flow, affinity chromatography with Cibarcron Blue 3GA Sepharose CL-4B and gel filtration with Sephadex G-100. The purity of rhNDPK-A was analyzed with SDS-PAGE and RP-HPLC. The Enzymatic activity was determined with RP-HPLC. The molecular weight (MW) was measured with matrix assisted laser desorption ionization time-of-flight MS (MALDI-TOF MS). The N-terminal residue was sequenced with Edman method. The apparent molecular weight of rhNDPK-A in solution was determined with multiangle laser light-scattering method (MALS). It was found that the purity of rhNDPK-A was 97.3% with SDS-PAGE method and 99.2% with RP-HPLC method. The specific enzymatic activity was (900 +/- 100) u/mg. The molecular weight was 17017, which was 132 less than the calculated value according to the amino acid sequence of NDPK-A. The sequencing result of rhNDPK-A revealed that its N-terminal residue was Ala, which was the second residue on N-terminal of native NDPK-A. The calculated MW of N-terminal deleted rhNDPK-A was 17017, exactly equal to the experimental value. The result of apparent MW determination revealed that rhNDPK-A formed homohexamer in solution with a MW of 102kD. These results suggested that rhNDPK-A possessed character identical to its native counterpart of assembling into hexamer. Confirming the identity of rhNDPK-A to its native counterpart provided a good foundation for drug development and mechanism study of NDPK-A.
Amino Acid Sequence ; Humans ; Molecular Sequence Data ; Molecular Weight ; NM23 Nucleoside Diphosphate Kinases ; chemistry ; isolation & purification ; metabolism ; Recombinant Proteins ; chemistry ; isolation & purification ; Scattering, Radiation ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

OBJECTIVETo investigate the effects of the enhancer of zeste homolog 2 (EZH2) gene on cell growth and invasion of the nasopharyngeal carcinoma (NPC).

METHODSRecombinant lentivirus vector for shRNA delivery of EZH2 was constructed and transfected into 293FT cells. After collecting the viral particles, the NPC cell line 5-8F cells were transfected. The effects of EZH2 silence on cell proliferation and cell cycle were detected using MTT assay, plate colony formation assay and flow cytometry. The migration and invasion of 5-8F cells were determined by wound healing assay and matrigel invasion assay, respectively. The expressions of EZH2 and epithelial-mesenchymal transition (EMT)-related markers at mRNA and protein levels were examined by real-time PCR and Western blot respectively.

RESULTSThe expressions of EZH2 mRNA and protein in the transfected 5-8F cells were obviously reduced. MTT assay showed that EZH2 downregulation significantly inhibited the growth of 5-8F/shEZH2 cells (P < 0.001). Colony formation rate (84.44%) of 5-8F/shEZH2 cells was lower than control (31.56%, P = 0.001). Cell cycle analysis showed that most 5-8F/shEZH2 cells were arrested in G0/G1 phase, with a very low ratio of cells in S phase. Wound healing assay indicated that the migration ability of cells silencing EZH2 decreased significantly, and the 48-hour relative migration distance of 5-8F/ShEZH2 cells and control cells was 0.58 ± 0.05, and 0.81 ± 0.02, respectively (P < 0.000). Matrigel invasion assay, showed the invasive capacity of cells silencing EZH2 was significantly inhibited, with less penetrating cells (72.23 ± 4.08) compared to control (150.95 ± 16.27), P < 0.000. The mRNA expressions of epithelial markers E-cadherin and Keratin 18 in the cells silencing EZH2 increased by 177% and 158% respectively, and the mRNA expressions of mesenchymal markers β-catenin and N-cadherin decreased by 18.04% and 41.18% respectively. Similar results also were obtained with Western blot analysis.

CONCLUSIONEZH2 significantly enhanced the proliferation and invasion of nasopharyngeal carcinoma cells in vitro, which might be mediated by inducing EMT.

Carcinoma ; Cell Line, Tumor ; Cell Proliferation ; Enhancer of Zeste Homolog 2 Protein ; Epithelial-Mesenchymal Transition ; Gene Expression Regulation, Neoplastic ; Humans ; Nasopharyngeal Neoplasms ; genetics ; pathology ; Neoplasm Invasiveness ; Polycomb Repressive Complex 2 ; genetics

This paper described a rapid and sensitive HPLC method to analyze (E)-3,5,4'-trimethoxystilbene (BTM-0512) in rat plasma and tissues. The analysis used a BDS Hypersil C18 analytical column (250 mm x 4.6 mm ID, 5 microm) and acetonitrile/water as the mobile phase. The UV detection wavelength was 319 nm. Proteins were precipitated with acetonitrile and diethylstilbestrol as internal standard. The method was validated according to State Food and Drug Administration of China and ICH of Technical Requirements for Registration of Pharmaceuticals for Human Use Guidelines. The limit of detection (S/N: 3/1) for BTM-0512 was 0.005 microg x mL(-1) for plasma. The method performances were shown to be selective for BTM-0512 and the linearity of the assay method was up to 10.0 microg x mL(-1) and 40.0 microg x g(-1) for plasma and tissues, respectively. At 0.1, 1 and 5 microg x mL(-1) (n=5), intraday and interday precision values (% RSD) were in the range of 2.6% - 5.1% and 2.4% - 4.8%, respectively. Mean accuracy and absolute recoveries of BTM-0512 ranged from 95.3% - 100.1% and 95.9% - 100.9% for plasma and tissues, respectively. This method can be quite useful for BTM-0512 pharmacokinetic and tissue distribution studies, for purpose which multiple plasma and tissue samples can be analyzed quickly with high reproducibility.
Angiogenesis Inhibitors ; blood ; pharmacokinetics ; Animals ; Anti-Allergic Agents ; blood ; pharmacokinetics ; Antineoplastic Agents ; blood ; pharmacokinetics ; Chromatography, High Pressure Liquid ; methods ; Male ; Rats ; Rats, Sprague-Dawley ; Reproducibility of Results ; Spectrophotometry, Ultraviolet ; methods ; Stilbenes ; blood ; pharmacokinetics ; Tissue Distribution

OBJECTIVETo study the epidemic pattern and trend of HBV infection in the area where the people had been immunized by HBV vaccine for 20 years.

METHODSThe whole sampling method was applied in combination with cross-sectional investigation. Blood samples were taken from every member of families. Markers of HBV infection were determined by using solid-phase radioimmunoassay (SPRIA).

RESULTS(1) The average HBsAg positive rate was 7.5%. The positive rate of markers for HBV infection of 0-19 years old subjects were lower than those of > or = 20 years old subjects. (2) The positive rate of HBsAg of 0-19 years old subjects in 1985 was higher than that in 2005. The anti-HBs positive rate in 1985 stemmed to be higher with age. It was 12.4% in 1- age group to 53.8% in >60 years age group. While the result of 2005 showed that the anti-HBs positive rate of 0-19 years old subjects dropped with age. The anti-HBc positive rate in 1985 also tended to be higher with age. But the result of 2005 showed that the rate of 0-19 years old subjects was just 1.4% to 16.8%.

CONCLUSIONThe epidemic patterns of HBV infection have had significant variations in the target population. HBV vaccine immunization has obtained excellent efficacy.

Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; China ; epidemiology ; Cross-Sectional Studies ; Female ; Hepatitis B ; blood ; epidemiology ; prevention & control ; Hepatitis B Vaccines ; administration & dosage ; therapeutic use ; Humans ; Immunization ; Infant ; Infant, Newborn ; Male ; Middle Aged ; Seroepidemiologic Studies ; Time Factors ; Treatment Outcome ; Young Adult

OBJECTIVETo study the epidemiological pattern and trends of hepatitis B virus (HBV) in the area where people had been immunized by HBV vaccine for long time.

METHODSThrough cluster sampling and cross-sectional study, relative information and blood samples from people in Long-an county by families were collected. Signals of HBV infection were tested by solid-phase reverse immunosorbent test.

RESULTS(1) The average HBsAg positive rate was 7.5% with anti-HBs as 44.5 %, and anti-HBc as 47.8%. The positive rates of HBsAg and anti-HBc among 0-19 year-olds were lower than those of > or = 20 year-olds. (2) The positive rates of HBsAg, anti-HBc and HBV infection among HBV vaccine immunized group were 2.8%, 12.0% and 12.5% respectively, comparing with which among the un-immunized group as 10.2%, 69.8% and 71.2% respectively. (3) The HBsAg positive rate of male was higher than the female's but with no significant difference of anti-HBs and anti-HBc between different sexes. (4) The average HBsAg positive rate of 0-19 years old group was only 2.4%, while that of 20-30 years old group was 13.6%-17.7% and dropped from 60 years old group and on. The anti-HBs positive rate of 0-19 years old people started to drop significantly by age. The anti-HBs and anti-HBc positive rates of > or = 20 years people were showing a rising trend by ages.

CONCLUSIONIt seemed obviously that the HBV epidemiological patterns had changed after HBV vaccine had been universally used for long time in newborns. The age peak of infection had been pushed backward for nearly 20 years. It had been proved that the HBV vaccine immunization program had obtained excellent efficacy.

Adolescent ; Adult ; Age Distribution ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; Female ; Hepatitis B ; epidemiology ; immunology ; prevention & control ; Hepatitis B Antibodies ; immunology ; Hepatitis B Surface Antigens ; immunology ; Hepatitis B Vaccines ; therapeutic use ; Humans ; Infant ; Infant, Newborn ; Male ; Middle Aged ; Young Adult

Obective To research and build an evaluation index system of emergency medical rescue capabilities (EMRC),and then put forward a new assessment method of EMRC.Methods A competency model of EMR was established by using Competency Model,then an evaluation index system was constructed on base of analyzing the factors of EMRC,and an evaluation model was set up based on four-element connection number of set pair analysis (SPA).Results The example showed that the calculation of the model was relatively simple,and the assessment results were to be believed,and moreover,its multiple functions could make up for the shortage of the single function of other evaluation models.Conclusion This method is scientific and rational to some extent,and can provide reference for similar evaluation problems.

Objective To investigate the nutrient contents of various grains in Hubei Province, and to provide a scientific basis for a balanced diet. Methods Various grains (except potatoes and beans) on the market in Hubei Province were collected. The samples were prepared by peeling and milling, or directly milling, and the nutrient contents were determined according to the national standard methods. With reference to the American nutrient profiling model, protein, dietary fiber, vitamin C, vitamin A, vitamin E, potassium, magnesium, iron and calcium, the nine nutrients were selected as recommended nutrients, and sodium, added sugars and fat were restricted nutrients. On the basis of 100 g for calculation, a nutrient-rich food model (Nutrient-rich foods, NRF_9.3) was established and applied to evaluate the grains. Results The evaluation results showed that wheat and buckwheat grains contained a higher recommended nutrient content than restricted nutrient content, and had higher nutritional value. Among them, quinoa (black) had the highest NRF index of 102.4, indicating the most nutritional value. After peeling, the loss rate of nutrient value (NRF_9.3 index) of various grains was 38.73% to 65.00%. Conclusion It is recommended that people should try to choose whole grains when purchasing grain products.