OBJECTIVETo explore the correlation between Pi and Shen by observing the relationship between the metabolism of aristolochic acid (AA) and mRNA and protein expression levels of organic anion transporting polypeptide (oatp) superfamily member 2a1 and 2 b1 (oatp2al and oatp2bl) in renal, small intestinal, and large intestinal tissues of Pi deficiency syndrome (PDS) model rats.

METHODSTotally 46 Sprague-Dawley (SD) rats were randomly divided into four groups, i.e., the blank group (n = 12), the PDS group (n = 22), the AA-I group (n = 6), and the PDS AA-I group (n = 6). PDS model was established by subcutaneously injecting Reserpine at the daily dose of 5 mg/kg for 16 successive days. Carotid intubation was performed in 6 rats selected from the blank group and the PDS group. Pharmacokinetics of AA-I were detected at 5, 15, 30, 45, and 60 min after gastrogavage of AA-I. AA-I concentrations in renal, small intestinal, and large intestinal tissues of 10 rats selected from the PDS group were determined. Normal saline was administered to 6 rats selected from the PDS group and the blank group by gastrogavage. Renal, small intestinal, and large intestinal tissues were collected in the AA-I group and the PDS AA-I group at 60 min after gastrogavage of AA-I. mRNA and protein expression levels of oatp2a1 and oatp2b1 in each tissue were detected using real-time polymerase chain reaction (RT- PCR) and Western blot.

RESULTSCompared with the blank group, plasma concentrations of in vivo AA-I were obviously higher in the PDS group at 15, 30, 45, and 60 min after gastrogavage of AA-I with statistical difference (P < 0.05). Plasma concentrations of AA-I were obviously decreased at 60 min after gastrogavage of AA-I; AA-I concentrations in renal and large intestinal tissues were elevated; AA-I concentrations in small intestinal tissues were obviously reduced in the PDS group. There was no statistical difference in mRNA expression levels of oatp2a1 and oatp2b1 in the aforesaid three tissues of rats between the blank group and the PDS group. Compared with the blank group, mRNA expression levels of oatp2a1 and oatp2b1 decreased in small intestinal tissues of the AA-I group, and the mRNA expression level of oatp2a1 in large intestinal tissues significantly decreased (P < 0.05, P < 0.01). Compared with the PDS group, mRNA expression levels of oatp2a1 and oatp2b1 increased in renal tissues of the PDS AA-I group (P < 0.05); mRNA expression levels of oatp2b1 increased in large intestinal tissues of the PDS AA-I group (P < 0.05).

CONCLUSIONSThe difference in AA-I metabolism might be associated with changed expression levels of oatp2a1 and oatp2b1 in renal, small intestinal, and large intestinal tissues under Pi deficiency induced loss of transportation. Shen and Dachang played important roles in substance metabolism under Pi deficiency state, which proved Pi-Shen correlated in Chinese medical theories.

Animals ; Anions ; Aristolochic Acids ; metabolism ; Drugs, Chinese Herbal ; Kidney ; Medicine, Chinese Traditional ; Organic Cation Transport Proteins ; metabolism ; Peptides ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley

Objective To establish the quality standard for Sanyuan Rupixiao Gel Paste. Methods Sparganii Rhizoma, Gleditsiae Sinensis Fructus, Cyperi Rhizoma and Impatientis Semen were identified by TLC method. The content of tetrahydropalmatine was determined by HPLC. Waters symmetry column was used with the mobile phase of acetonitrile-0.1% phosphatic acid in a gradient manner (pH was adjusted to 6.4 by triethylamine) (55:45) at the detection wavelength of 280 nm. The flow rate was 1.0 mL/min at the column temperature of 30 ℃. Results The spots in TLC were clear without any interference;tetrahydropalmatine showed a good linear relation in the range of 0.092–1.84 μg;the average recovery was 100.15%with RSD of 1.58%(n=6). Conclusion The method is simple and accurate with high reproducibility, which can be used for the quality control of Sanyuan Rupixiao Gel Paste.

OBJECTIVETo study the distribution and drug resistance of the isolated bacteria from children with acute respiratory infection (ARI) in Dalian.

METHODSBetween January 2006 and February 2007, 930 children with ARI were enrolled, including 364 with acute upper respiratory infection (AURI), and 566 with acute lower respiratory infection (ALRI). The AURI children, who did not receive antimicrobial agent treatment or received oral antimicrobial agents 1-2 times, had bacterial cultures of pharyngeal swab. The ALRI children, who received intravenous antibacterial agents more than 3 days, had bacterial cultures of sputum and bronchoalveolar lavage fluid (BALF). Isolated bacteria were identified by the ATB system (Bio-Merieux, France). Antimicrobial susceptibility testing was carried out by means of Kirby-bauer.

RESULTSA total of 404 isolates (43.4%) were identified. Haemophilus influenzae, Haemophilus parainfluenzae and Streptococcus pneumoniae accounted for 22.5%, 12.1% and 7.4% respectively. In the isolates from AURI, Haemophilus influenzae, Haemophilus parainfluenzae and Streptococcus pneumoniae accounted for 43.9%, 22.0% and 9.1% respectively; Escherichia coli, Klebsiella pneumonia and Nonfermenters accounted for 4.5%, 8.3% and 3.0% respectively. In the isolates from ALRI, Haemophilus influenzae, Haemophilus parainfluenzae and Streptococcus pneumoniae accounted for 12.1%, 7.4% and 6.6% respectively; Escherichia coli, Klebsiella pneumoniae and Nonfermenters accounted for 16.9%, 13.2% and 21.8% respectively. The resistant rates of Haemophilus to ampicillin and TMP-SMZ were 29.3% and 32.9% respectively, and to amoxicillin-clavulanic acid, cefalotin, cefaclor, cefuroxime and cefotaxime were 12.1%, 10.0%, 10.0%, 11.4% and 5.7%, respectively. The resistant rate of Haemophilus to ampicillin, amoxicillin-clavulanic acid, cefaclor, tetracycine and TMP-SMZ in the ALRI group were significantly higher than that in the AURI group (P<0.05 or 0.01).

CONCLUSIONSIn Dalian, Haemophilus was the main isolate of children with ARI. The distribution of bacteria was different between ALRI and AURI. In ALRI, Gram-negative bacilli were in a higher proportion, and the resistant rates of Haemophilus influenzae and Haemophilus parainfluenzae to ampicillin, amoxicillin-clavulanic acid and cefaclor were higher.

Acute Disease ; Adolescent ; Bacteria ; drug effects ; Child ; Child, Preschool ; Drug Resistance, Bacterial ; Female ; Humans ; Infant ; Male ; Respiratory Tract Infections ; drug therapy ; microbiology

Objective　To observe some biological features of bone marrow mesenchymal stem cells and explore the best conditions for isolatin g and culturing in vitro. Methods　Common cell culture techn ique, light and electron microscopy were used to study the effects of the growth , proliferation, morphology of the bone marrow mesenchymal stem cells in differe nt adherent time, concentration of serum and cell density. Results　The best culture condition in vitro for growth was 4-24 hours adherent time, 5%-10% fetal bovine serum, (4-8)×104/ml cell density. The cells were sp indle in shape and had a strong ability of proliferation. The time for cell duplication was 3 to 4 days. The cells showed the characteristics of stem cell s in electron microscope. Conclusion　The best condition for iso lation and culture of bone marrow mesemchymal stem cells was successfully establ ished and some biological features were obserred. It found a base for further in vestigation and using of mesenchymal stem cells.

China ; epidemiology ; Humans ; Smoking ; epidemiology ; Smoking Cessation ; Smoking Prevention

Histone lysine methyltransferase EZH2 has been reported to be frequently overexpressed in hepatocellular carcinoma (HCC) tissues and associated with hepatocarcinogenesis. However, the exact mechanism of EZH2 up-regulation in HCC has not been determined. In this study, we used murine hepatocyte AML12 cells to investigate the role of hepatitis B virus X protein (HBx) in regulating the expression of mEZH2. Western blot analysis demonstrated that the expression level of mEZH2 protein in AML12 cells was up-regulated by HBx in a dose-dependent manner. To further investigate the mechanism of mEZH2 overexpression, the 2500 bp regulatory sequence upstream from the first exon of the mEZH2 gene was amplified from AML12 genomic DNA and constructed into a luciferase reporter plasmid. The luciferase activity of the mEZH2 promoter significantly increased in AML12 cells co-transfected with HBx plasmid, and deleting the -486/-214 promoter region decreased HBx-induced mEZH2 promoter activation by nearly 50%. The -486/-214 region was then analyzed in the TRANSFAC 6.0 database and a typical E2F1-binding site was found. Mutation of this E2F1-binding site or knockdown of E2F1 expression by RNAi led to a dramatic decrease in HBx-induced activation of the mEZH2 promoter and mEZH2 overexpression in AML12 cells. These results provide evidence that HBx up-regulates mEZH2 expression by transactivating the mEZH2 promoter through E2F1 transcription factor, thereby providing new epigenetic evidence for the carcinogenic effect of HBx.
Animals ; Binding Sites ; Cell Line ; E2F1 Transcription Factor ; genetics ; Enhancer of Zeste Homolog 2 Protein ; Hepatocytes ; cytology ; metabolism ; virology ; Histone-Lysine N-Methyltransferase ; genetics ; metabolism ; Mice ; Plasmids ; Polycomb Repressive Complex 2 ; Promoter Regions, Genetic ; genetics ; RNA, Small Interfering ; genetics ; Trans-Activators ; genetics ; metabolism ; Transfection ; Up-Regulation

Using SDS-PAGE analysis, the High-molecular-weight (HMW) glutenin subunits of two Crithopsis delileana accessions were detected. It was found that the two accessions had the same HMW glutenin subunits. Only one HMW glutenin subunit with the similar electrophoresis mobility to the y-type HMW glutenin subunit of hexaploid wheat was observed in C. delileana. However, It was indicated that this glutenin subunit was an x-type glutenin subunit Kx by gene sequence analysis. The full coding region of Kx gene is 2052bp and could encodes a mature protein with 661 amino acid residues. The Kx gene could be expressed in the bacterial expression system, and the expressed protein had the same electrophoresis mobility as that in the seed of C. delileana. The primary structure of Kx subunit was very similar to the x-type HMW glutenin subunits encoded by the A, B and D genomes of wheat, the C and U genomes of Aegilops, and the R genome of Secale cereale. In the phylogenetic analysis, Kx subunit was clustered together with Ax1 subunit by an interior parallelled branch. In conclsion, Kx gene is a novel x type glutenin subunit gene from C. delileana.
Amino Acid Sequence ; Base Sequence ; Genes, Plant ; Glutens ; genetics ; metabolism ; Molecular Sequence Data ; Plant Proteins ; genetics ; metabolism ; Poaceae ; genetics ; metabolism ; Triticum ; genetics ; metabolism

Objective To investigate the expressions of survivin, cyclooxyenase-2 (COX-2) and vascular endothelial growth factor (VEGF) and their relationship with angiogenesis in condyloma acuminatum (CA) tissues. Methods Immunohistochemistry using PowerVision staining kit was performed to detect the expression of survivin, COX-2 and VEGF protein in 60 CA tissue samples from patients and 21 normal skin samples from the foreskin of human controls. At the same time, the microvessel density was determined in CA tissues by staining blood vessel endothelium with anti-CD105 monoclonal antibody. Results The positivity rate of survivin and COX-2 expression was 56.67% and 63.33%, in CA tissues, 9.52% and 0 in normal skin tissues, respectively. There was a significant difference between the two groups of tissue samples in the positivity rate and intensity of survivin and COX-2 expression (all P < 0.05). VEGF was expressed in all of the CA tissues and normal skin tissues, while the intensity of VEGF expression was statistically different between the two groups of tissue samples (P < 0.05). The MVD was 16.38 ± 5.46 and 0.62 ± 0.44 in CA tissues and normal skin tissues, respectively (P < 0.05). There was a significant positive correlation between the expressions of survivin, COX-2 and VEGF, as well as between MVD and the expressions of survivin and COX-2 in CA tissues. Conclusion The expression levels of survivin, COX-2 and VEGF are significantly higher in CA tissues than in normal skin tissues.

OBJECTIVETo investigate the correlation of sexual dysfunction with psychological abnormalities by analyzing the sexual function and mental state of the patients with redundant prepuce or phimosis.

METHODSThis study included 216 randomly selected patients with redundant prepuce or phimosis and 85 normal male controls. We conducted investigations among the subjects using a questionnaire on the general data of the patients, Chinese Index of Sexual Function for Premature Ejaculation (CIPE), International Index of Erectile Function (IIEF), and Symptom Checklist 90 (SCL-90). Then we assessed the status of premature ejaculation (PE) and erectile dysfunction (ED), calculated the incidence of psychological abnormalities, such as depression and anxiety, and analyzed the correlation of PE and ED with the mental state of the patients.

RESULTSThe PE and ED patients scored significantly higher than normal controls on SCL-90, somatization, compulsion, depression, anxiety and other factors (P < 0.05). CIPE scores were correlated with the scores on SCL-90, somatization, compulsion, interpersonal sensitivity, depression, anxiety and other factors, while ED-related scores showed no correlation with the scores on SCL-90 and other factors.

CONCLUSIONPatients with redundant prepuce or phimosis have poor mental health, and there is an interaction between PE and the mental state of the patient.

Adolescent ; Adult ; Case-Control Studies ; Erectile Dysfunction ; etiology ; Humans ; Male ; Middle Aged ; Phimosis ; physiopathology ; psychology ; Sexual Behavior ; Sexual Dysfunction, Physiological ; physiopathology ; psychology

OBJECTIVETo evaluate the roles of cell adhesion, multidrug resistance and cell proliferation in short-term recurrent cases with superficial bladder cancer, and the prognostic value of the three indexes.

METHODSImmunohistochemical staining for E-cad, P-gp and Ki-67 was performed on the tumors of 100 patients with stage T0-T1 transitional cell carcinoma of the bladder who had been included in a retrospective research by follow-up.

RESULTSE-cad and P-gp expression was positive in 51 (43.2%)and 17 (14.4%) of the tumors, respectively and mean proliferation index (PI) was 22.1%. The decrease in E-cad expression was accompanied with the increasing recurrent episodes (P < 0.05), while increase of P-gp expression and PI were accompanied with the increasing recurrence episodes (P < 0.05). There was significant difference according to E-cad, P-gp positivity and between T(1)G(3) patients and no-T(1)G(3) patients (P < 0.05). There was negative correlation of E-cad expression with P-gp expression and PI.

CONCLUSIONSMinimum adhesion, strong drug resistance and maximum proliferation are the main factors that promote short-term recurrence of superficial bladder cancer and also the inherent reasons for easy recurrence and high malignancy of T(1)G(3) tumors. During this course, the three aspects may interact.

ATP-Binding Cassette, Sub-Family B, Member 1 ; analysis ; Adult ; Cadherins ; analysis ; Cell Adhesion ; Cell Division ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; Female ; Humans ; Ki-67 Antigen ; analysis ; Male ; Neoplasm Recurrence, Local ; etiology ; Urinary Bladder Neoplasms ; drug therapy ; etiology ; pathology