OBJECTIVE To investigate and compare the enzyme kinetic characters of psoralen (PRN)and isopsoralen(IPRN)in rat and human liver microsomes. METHODS PRN and IPRN in liver microsomes incubates were determined using LC-MS/MS. The enzyme kinetic and metabolic stability of PRN and IPRN were investigated by employing the optimized rat and human liver microsomes incubations. The Vmax and Km values were calculated using the nonlinear regression method. RESULTS The quanti?tative method showed good linearity within the range of 0.1-50.0 μmol · L-1 and was suitable for the assay in biological samples. The in vitro elimination was linear with the substrate concentrations lower than 1 μmol,the protein concentration within 0.5 g · L-1,and the incubation time within 40 min. The t1/2 values of PRN and IPRN in rat and human liver microsomes were 74.5,95.0,74.5 and 173.3 min, respectively. The Vmax values of PRN in rat and human liver microsomes were(1.140±0.080)μmol·min-1·g-1 protein,(0.620±0.060)μmol·min-1·g-1 protein,while Km values of PRN in rat and human liver microsomes were (12.9 ± 0.3)μmol · L- 1,(7.4 ± 1.3)μmol · L- 1,respectively. The Vmax values of IPRN in rat and human liver microsomes were(0.251±0.012)and(0.103±0.014)μmol·min-1·g-1 protein,while Km values of IPRN in rat and human liver microsomes were (3.0 ± 0.4)μmol · L-1,(3.4 ± 0.7)μmol · L-1,respectively. CONCLUSION The enzyme kinetic characters and metabolic stability of PRN and IPRN show species and chemical structures related differences. Interestingly,the metabolic eliminations of PRN and IPRN are similar in rats. However,the metabolic elimination of IPRN in humans involved in CYP enzymes may be much slower than that of PRN.

Objective: To investigate the expression and identification of CH50 polypeptide in bladder cancer cell line BIU-87 by gene transfection. Methods:The eukaryotic expressing vector pCH510 of polypeptide CH50 was introduced into BIU-87 cells by gene transfection Lipofectimine TM2000.The expressed product was identified by immunohistochemistry and Western blot method. The expression of the transfected gene was identified by RT-PCR. Results:The BIU-87 cells could produce CH50 polypeptide by transfection. Conclusion:When the vector of pCH510 was transfected into BIU-87 cells in vitro, the cell adhension characteristic would be changed.

Objective To investigate the effect of HSP70/CD80 DNA vaccine on the airway inflammation and hyperresponsiveness of asthmatic mice. Methods Forty female healthy BALB/c mice were randomly divided into 4 groups; saline group, asthma group, pcDNA3. 1 plasmid control group, and prevention group with HSP70/CD80 DNA vaccine, with 10 mice in each group. The mice were immunized by intramuscular( i. m. ) injection with HSP70/CD80 DNA vaccine before sensitization and challenge with ovalbumin. Then, the murine model of allergic asthma was made with injection of ovalbumin intraperitoneal ( i. p. ) , and inhalation of ovalbumin. Before mice were sanctified, their airway hyperresponsiveness( AHR) was measured. After mice were sanctified, bronchoalveolar lavage fluid( BALF) was obtained and cytokine IL-13 and IFN-γ were measured. And the lung histology and histochemistry were examined. Results Compared with mice in asthma and pcDNA3. 1 group, mice in vaccine group showed significantly reduced airway inflammation (P＜0. 05) and AHR (P＜0. 05). IFN-γ content in BALF were increased in mice from vaccine group compared with the asthma group and the pcDNA3. 1 group ( P ＜0. 05) , and IL-13 content in BALF were decreased. Conclusion HSF70/CD80 DNA vaccine can reduce airway inflammation and hyperresponsiveness in asthmatic mouse and this chimerical plasmid could be a candidate vaccine to prevent asthma.

OBJECTIVETo study the predicting effect of proly 4-hydroxylase beta polypeptide (P4HB) in treating non-small cell lung cancer (NSCLC) patients by Yiqi Chutan Recipe (YCR).

METHODSTotally 46 stage III and IV NSCLC patients were treated by YCR for 4 therapeutic courses. Effect was assessed by RECIST of solid tumor. P4HB expression was detected in the lung cancer tissue by immunohistochemical assay. Factors affecting disease control rates (DCR) of YCR were analyzed by Logistic regression analysis. The correlation between P4HB expression and the effect of YCR was analyzed.

RESULTSThe DCR of advanced NSCLC treated by YCR was 36.96% (17/46 cases). P4HB was high expressed in advanced lung cancer tissue (6/15 cases). Gender, initial treatment, and retreatment are independent factors for affecting DCR of treating lung cancer by YCR.

CONCLUSIONP4HB might be taken as a factor for predicting the effect of YCR in treating NSCLC.

Carcinoma, Non-Small-Cell Lung ; drug therapy ; metabolism ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Lung ; Lung Neoplasms ; drug therapy ; metabolism ; Male ; Procollagen-Proline Dioxygenase ; metabolism ; Protein Disulfide-Isomerases ; metabolism

Objective To study effects of Rupixiao granules on contraction of uterus of mice in vitro. Methods Mouse model with isolated uterine contraction was established. The mice pretreated with estradiol benzoate were sacrificed and then the uteri were taken out. The normal contractions were recorded by the biological function system. Effects of 10, 20, 30, 40, 50, 60 mg.mL-1 Rupixiao granules on spontaneous and oxytocin-induced (5 U.L-1) uterine contractions were recorded,inhibition of Rupixiao granules on mean constriction amplitude,constriction frequency and contraction activity of pitocin-pretreated uterus was observed and the inhibition rates were calculated. Results Compared with the baselines, the amplitude, frequency, activity of spontaneous uterine contraction were significantly down-regulated by Rupixiao solution at a range of 10-60 mg.mL-1 ,as well as it could inhibit oxytocin-induced spasmodic contraction of isolated uterus at 10-50 mg . mL-1 ( P<0. 01 ) . Conclusion The Rupixiao granules inhibits spontaneous contraction of isolated mouse uteri and oxytocin-induced spasm of uterine smooth muscle in vitro.

Objective To evaluate the feasibility and therapeutic efficacy of real-time virtual navigation system (RVS) combined with contrast-enhanced ultrasonography (CEUS) for hepatocellular carcinoma undetectable by conventional ultrasound and CEUS.Methods A total of 213 patients with 265 lesions (undetectable on conventional ultrasound but detectable by other imaging examinations) were enrolled in this study.CEUS was performed,and lesions which were detected with CEUS were given CEUS guided radio-frequency ablation (RFA) (CEUS group).RVS combined with CEUS was performed to locate the rest lesions and guide RFA (RVS + CEUS group).Diagnostic value and therapeutic efficacy of RVS + CEUS were assessed.Results In 256 lesions,174 (65.7％) could be detected with CEUS,and among the rest 91 lesions,82 (90.1％) lesions could be detected by RVS combined with CEUS.The technique effectiveness rate of RVS + CEUS was significantly higher than that of CEUS (x2 =18.85,P ＜0.0001).Complete ablation rate after one month in RVS+ CEUS group was 9t.5％ (75/82),which was significantly higher than that in CEUS group [86.2％ (150/174),P ＜0.05].Local progression in one year in CEUS group and RVS + CEUS group were 13.4％ and 10.9％,respectively.Conclusions RVS combined with CEUS is feasible for hepatocellular carcinoma undetectable by conventional ultrasonography and CEUS,and the therapeutic efficacy of RVS + CEUS guided RFA is satisfactory.

Objective To determine the curative effect of dilation for achalasia with temporary cardia stent in different diameters based on a long-term follow-up.Methods The study cohort was comprised of 135 patients of achalasia.Among them differentiated by stent diameters as followings:30 patients were treated under fluoroscopy with dilation of temporary cardia stent in 20 mm diameter(group A), 30 patients with dilation of temporary eardia stent in 25 mm diameter(group B),and 75 patients with dilation of temporary cardia stent in 30 mm diameter(group C).135 cardia stents were temporarily placed in the 135 patients and withdrawn after 3 -5 days via gastroscopy.All the stents were inserted and withdrawn successfully.The follow-up in all groups lasted 6-128 months.Results Six(20.0%)out of 30 patients,6(20.0%)out of 30 patients,5(22.7%)out of 22 patieuts,6(37.5%)out of 16 patients,5 out of 9 patients,3 out of 3 patients in group A exhibited dysphagia relapse during 6 months,1 year,3 years,5 years,8 years,and 10 years follow-up,respectively. Four(13.3%)out of 30 patients,4(13.3%)out of 30 patients,3(13.0%)out of 23 patients,4(22.2%)out of 18 patients,5(45.5%)out of 11 patients,and 3 out of 4 patients in group B exhibited dysphagia relapse during 6 months,1 year,3 years,5 years,8 years,and 10 years follow-up,respectively.No(0.0%)out of 75 patients, 1(1.5%)out of 66 patients,4(8.3%)out of 48 patients,6(18.2%)out of 33 patients,6(33.3%)out of 18 patients,2 out of 5 patients in group C exhibited dysphagia relapse during 6 months,1 year,3 years,5 years,8 years,and 10 years follow-up,respectively.Conclusion Dilation with temporary cardia metal stent in 30 mm diameter is the best dilation for achalasia in long-term follow-up.

OBJECTIVETo explore the clinical features of horizontal semicircular canal benign paroxysmal positional vertigo.

METHODSThe clinical manifestations of 239 patients with horizontal semicircular canal benign paroxysmal positional vertigo from August 2003 to December 2010 were retrospectively analyzed.

RESULTS25.7% (239/931) of all the benign paroxysmal positional vertigo patients were the horizontal semicircular canal benign paroxysmal positional vertigo was involved. One hundred and ninety-seven patients showed geotropic nystagmus in head rolling test with a mean latency period of (0.88 ± 0.72) s and a mean duration period of (26.36 ± 19.71) s. Forty-two patients showed apogeotropic nystagmus in head rolling test with a mean latency period of (2.69 ± 1.83) s and a mean duration period of (53.48 ± 43.12) s. Among all the horizontal semicircular canal benign paroxysmal positional vertigo patients, 39 (16.3%) presented horizontal nystagmus with slight upbeating component. The nystagmus latency in apogeotropic nystagmus group was longer than that in geotropic nystagmus group (t = -6.33, P < 0.001), and nystagmus duration period was also longer (t = -3.99, P < 0.001). Applied Barbecue maneuver to the patients with geotropic nystagmus, 192 patients were cured after (1.6 ± 0.8) rotations; After applied head shaking maneuver to the patients with apogeotropic nystagmus, 40 patients changed to geotropic nystagmus and cured after (1.9 ± 0.8) rotations.

CONCLUSIONSIncidence rate of horizontal semicircular canal benign paroxysmal positional vertigo was higher than expectation. Barbecue maneuver was applied to patients with geotropic nystagmus. While to the patients with apogeotropic nystagmus, head shaking maneuver should be performed firstly and then followed by Barbecue maneuver.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Benign Paroxysmal Positional Vertigo ; Female ; Humans ; Male ; Middle Aged ; Nystagmus, Pathologic ; Retrospective Studies ; Semicircular Canals ; Vertigo ; diagnosis ; therapy ; Young Adult

To investigate the effect of huangqin tang on expression of cytokines and NF-κB p65 in rats with ulcerative colitis (UC), and to probe into its underlying mechanisms of action. The mode of UC rats with cell immunoreactivity was made using compound method (trinitrobenzene sulfonic acid and ethanol). Rats were randomly divided into control group, model group, SASP group and high dose, middle dose and low dose of huangqin tang group. The food intake, body weight and microscopic damage of rats in each group were evaluated after being treated for five days. The blood and colon tissue were also collected. Production of NO was detected by Griess assay, the expression levels of IL-6, TNF-α, PGE2 were detected by ELISA. ICH method was undertaken to determine the expression of NF-κB p65 protein in colon tissue. The food intake and body weight of model group rats were lower than that of control group. The expression levels of NO, IL-6, TNF-α, PGE2 in serum and NF-κB p65 protein of colon tissue in model group were higher than that of control group. The above indexes were ameliorated in high and middle dose of huangqin tang groups. But there was no significant difference with SASP group. NF-κB p65 may be involved in the pathogenesis of UC, and huangqin tang can inhibit the relative activity of NF-κB p65, and decrease the expression levels of NO, IL-6, TNF-α and PGE2.
Animals ; Colitis, Ulcerative ; metabolism ; Dinoprostone ; blood ; Drugs, Chinese Herbal ; pharmacology ; Interleukin-6 ; blood ; Nitric Oxide ; blood ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; drug effects ; Transcription Factor RelA ; metabolism ; Tumor Necrosis Factor-alpha ; blood

The goal of this study is to research the genetic characteristics and relationship between HN and P genes of NDV. The nucleotide sequence and deduced amino acid sequence were analyzed for the Hemagglutinin-neuramindase (HN) and Phosphoprotein (P) gene of twelve field isolates of Newcastle disease virus (NDV) during 1997-2005 in China. The HN and P gene sequences of fifteen NDV reference strains from GenBank were also used in this study. The molecular evolution distance of nucleotides and amino acids were calculated by MEGA 4.0 software, and analysis of variance and correlations were analyzed by SPSS11.0 software among different length sequences of the HN gene or P gene. The nucleotide and amino acids correlation of HN and P gene were analyzed respectively. The correlation of evolution distance and isolation year were also calculated. The results indicated that there were difference and good correlation of nucleotide and amino acid among different length sequences of the HN gene or P gene. These results revealed that the HN and P gene of NDV have the different response to selective pressure to adopt to landscape and closely relationship on heredity mutations. Nucleotide variations of HN and P gene have relationship with isolation year of strains.
Evolution, Molecular ; Genetic Variation ; HN Protein ; genetics ; Newcastle disease virus ; classification ; genetics ; Phosphoproteins ; genetics ; Reverse Transcriptase Polymerase Chain Reaction