K21. The mean t1/2(?) was (2.7?0.4) h, Vd was about 11.18 L?kg-1.The C-T profile conformed to two compartment open model. The plasma Dau concentration-time curves showed a double-peak phenomenon in all dosages of all dogswhen dauricine was given by intragastric was.The tpeak(1) was (0.8?0.6) ～(1.2?0.5) h,tpeak(2) was (5.2?3.2) ～(6.5?1.9)h,Cmax(2) 0.05) and the AUC was increased in proportion.The drug is eliminated non-linearly when the dosage is above 50 mg?kg-1, the parameters t1/2(el),CL, AUC/X0 shows great difference (P

Aim The relative bioavailability of domestic ibudilast sustained release capsules in healthy volunteers was observed.Methods A single oral dose of 20 mg of imported and domestic ibudilast sustained release capsules and 10 mg of ibudilast raw material was separately given to 12 healthy volunteers in a randomized crossover study. Ibudilast concentration in plasma was determined by HPLC method.Results The Cmax were (54.9?9.7),(60.7?9.1) and (62.2?11.5) ?g?L-1; the tmax were (3.8?0.8),(3.9?0.8) and (1.8?0.3) h;the t1/2(ke) were (1.5?1.4),(12.1?1.0) and (3.5?0.5) h,and the AUC(0～t) were (618.1?57.7),(588.1?66.6) and (233.0?46.4) ?g?h?L-1 in imported capsule group, domestic capsule group and raw material group respectively. The relative bioavailability of domestic sustained release capsules of ibudilast is (95.6?11.0)%. Conclusion The results of statistical analysis demonstrate that the imported and domestic sustained capsules have significant character of significantly sustained release and are bioequivalent.

Objective To observe the efficacy and safety of in-center nocturnal hemodialysis (INHD) in uremic patients. Methods Thirty-two maintenence hemodialysis (MHD) patients received INHD (3 times per week and 7.5 hours each session) for 6 months.Before and 1, 3 and 6 months after entering INHD, blood routine, hepatic and renal function,serum electrolyte, lipids, parathyroid hormone and β2-microglobulin(β2-MG) were assayed, Kt/V and URR were calculated. Blood pressure of each dialysis session 2 months before and 6 months after INHD was recorded. Cardiac ultrasound and SF-36 questionnaire before and after INHD were performed. Use of drugs was recorded. Results Compared with 2 months before INHD, predialysis BP decreased [(130.3/86.0) vs (139.3/88.6) mm Hg, P＜0.01], while post-dialysis BP raised significantly [(121.1/80.5) vs (115.0/77.8) mm Hg, P＜0.01] 6 months after INHD.Intradialysis hypertension (9.8%vs 24.0%) and hypotension (7.3% vs 14.9%) both reduced (all P＜0.01). Serum phosphorus [(1.37±0.27) vs (2.08±0.49) mmol/L, P＜0.01] and iPTH [(355.4±139.6) vs (632.3±750.0) ng/L, P＜0.01] decreased, while calcium increased [(2.64±0.25) vs (2.28±0.37) mmol/L, P＜0.01], HDL[(1.27±0.29) vs (0.75±0.08) mmol/L] increased, LDL [(2.04±0.52) vs (2.75±0.75) mmol/L] decreased (all P＜0.05). URR [(79.7±0.1)% vs (64.7±4.7)%] and Kt/V (1.40±0.44 vs 0.89±0.25, P＜0.01) increased. Serum β2-MG decreased [(17.3±3.9) vs (24.6±5.9) mg/L, P＜0.01]. LVMI decreased [(99.8±29.0) vs (114.8±72.7), P＜0.05]. Physical functioning, role-physical and role-emotional of SF-36 increased (all P＜0.01). The types of antihypertension drug, dosage of EPO, Vitamin D3 and phosphorus binder decreased (all P＜0.01).Patients of drug withdrawal increased (P＜0.05). Conclusion The hypertension, anemia,calcium-phosphorus metabolism, lipid disorder, cardiac malfunction and the quality of life are improved in INHD patients.

OBJECTIVEThe aim of this study was to explore the possibility of creating a toxin, C-CPE-ETA', by fusing C-terminal high affinity binding domain of CPE (C-CPE) with a truncated form of Pseudomonas aeruginosa exotoxin A (ETA') and to examine whether C-CPE-ETA' could specifically target CLDN-3, 4 molecule and the targeted toxin was cytotoxic against CLDN-3,4-overexpressing ovarian cancer.

METHODSCLDN-3 and CLDN-4 expressions were analyzed at the mRNA level in three ovarian cancer cell lines and epithelial ovarian cancer tissues from 20 patients. After transforming an expression plasmid of C-CPE-ETA' into E. coli BL21 (DE3) plysS strain, the recombinant protein was purified using His-Bind resin chromatography column and analyzed by Western blot and Coomassie blue staining. The specific binding, proapoptotic and cytolytic activities were evaluated by flow cytometry, fluorescence microscopy with the JC-1 probe and MTT assay in CLDN-3,4-overexpressing ovarian cancer cells.

RESULTSQuantitive RT-PCR results showed there existed high levels of CLDN-3 and CLDN-4 in ovarian cancer cells, CAOV3, OVCAR3 and SKOV3. Moreover, high expressions of CLDN-3 and CLDN-4 were observed in 90.0% (18/20) and 60.0% (12/20) of ovarian cancer tissues, with an expression level 10-fold higher than that in the normal ovarian tissue. A 58 000 recombinant protein C-CPE-ETA' was demonstrated by Western blot and Coomassie blue staining. Purified and recombinant C-CPE-ETA' was bound with high affinity to CLDN-3,4-overexpressing ovarian cancer cells, CAOV3, OVCAR3 and SKOV3 cells. C-CPE-ETA' was strongly proapoptotic and cytotoxic towards the CLDN-3,4-overexpressing ovarian cancer cells. The concentration of IC(50) was 7.364 ng/ml for CAOV3 cells, 8.110 ng/ml for OVCAR3 cells and 22.340 ng/ml for SKOV3 cells, respectively. However, control CLDN-3,4-deficient cell line HUVEC was not susceptible to the recombinant C-CPE-ETA' at a concentration up to 10 µg/ml.

CONCLUSIONSThe C-CPE-ETA' protein exhibits remarkably specific cytotoxicity for CLDN-3,4-overexpressing ovarian cancer cells. Its therapeutic potential warrants further development for ovarian cancer molecular targeted therapy.

ADP Ribose Transferases ; metabolism ; physiology ; Apoptosis ; Bacterial Toxins ; metabolism ; Cell Line, Tumor ; Claudin-3 ; Claudin-4 ; Claudins ; genetics ; metabolism ; Enterotoxins ; metabolism ; physiology ; Exotoxins ; metabolism ; physiology ; Female ; Humans ; Immunotoxins ; metabolism ; Ovarian Neoplasms ; metabolism ; pathology ; RNA, Messenger ; metabolism ; Recombinant Fusion Proteins ; metabolism ; physiology ; Virulence Factors ; metabolism ; physiology

OBJECTIVETo detect the mutations of autosomal dominant polycystic kidney disease gene 2(PKD2)in Chinese.

METHODSThe white blood cell genomic DNA from patients of 94 Chinese autosomal dominant polycystic kidney disease(ADPKD) pedigrees was isolated and amplified by polymerase chain reaction(PCR). The PCR products were analyzed by denaturing high-performance liquid chromatography(DHPLC). The samples with abnormal profiles were sequenced.

RESULTSEight mutations were identified, including 2 nonsense mutations, 2 deletion mutations,1 insertion mutation and 3 missense mutations. Two nonsense mutations occurred in exon 5(1249C-->T) and exon 13(2407C-->T),both resulted in a stop codon. The insertion was in exon 2(636-637 ins T),and the deletion mutations were in exons 12(2348-2351 del AGAA) and 13(2401 delete A),resulting in the reading frame shift. Three missense mutations were in exons 1(G568-->A),4(C964-->T),and 5(G1168-->A), which caused amino acid changes (190Ala-->Thr,322Arg-->Trp,390Gly-->Ser).

CONCLUSIONThe method of DHPLC was used in detecting mutations successfully and 8 mutations in PKD2 were identified. It will be useful in the molecular diagnosis of ADPKD in advance of the cysts formation and birth.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Chromatography, High Pressure Liquid ; Female ; Humans ; Male ; Membrane Proteins ; genetics ; Middle Aged ; Mutation ; Nucleic Acid Denaturation ; Polycystic Kidney, Autosomal Dominant ; genetics ; TRPP Cation Channels

Cell Line ; Collagen Type III ; metabolism ; Gene Expression Regulation ; Gene Knockdown Techniques ; Genetic Vectors ; Hepatic Stellate Cells ; metabolism ; Humans ; Liver Cirrhosis ; pathology ; prevention & control ; Plasmids ; genetics ; RNA, Messenger ; genetics ; metabolism ; RNA, Small Interfering ; Signal Transduction ; Smad4 Protein ; genetics ; metabolism ; Transfection ; Transforming Growth Factor beta ; metabolism

Adolescent ; Adult ; Aged ; Antithyroid Agents ; therapeutic use ; Female ; Graves Disease ; blood ; drug therapy ; Humans ; Iodides ; urine ; Iodine ; administration & dosage ; Male ; Middle Aged ; Propylthiouracil ; therapeutic use ; Prospective Studies ; Sodium Chloride, Dietary ; administration & dosage ; Thyroid Hormones ; blood

OBJECTIVETo explore genes involved in chronic myeloid leukemia (CML) with t (3; 21) (q26; q22) chromosome translocation in blastic crisis.

METHODSA case of CML patient with t (3; 21) (q26; q22) in blastic crisis was reported. AML1 and bcr/abl genes were detected by FISH in interphase and metaphase cells. Genes involved in t (3; 21) (q26; q22) were analysed by RT-PCR and sequencing.

RESULTSAML1 gene hybridization signal was detected in der (3) and der (21) chromosomes. AML1-Evi1, AML1-MDS1-Evi1, AML1-EAP fusion transcripts and Evi1 gene were detected in mRNA level, but no AML1-Evi1 fusion transcript. The mRNA expression level of AML1-MDS1-Evi1 fusion gene was 1.58 and 1.54 times higher than that of AML1-MDS1 and AML1-EAP, respectively. The mRNA expression level of Evi1 gene of the patient was 2.71 times higher than that of HEL cell line.

CONCLUSIONt (3; 21) (q26; q22) resulted in the AML1-MDS1-Evi1, AML1-MDS1, AML1-EAP fusion transcripts, and Evi1 gene was also activated by the translocation. These secondary aberrations should be the molecular basis of CML patient with t (3; 21) (q26; q22) in blastic crisis.

Adult ; Blast Crisis ; genetics ; pathology ; Chromosomes, Human, Pair 21 ; genetics ; Chromosomes, Human, Pair 3 ; genetics ; Core Binding Factor Alpha 2 Subunit ; genetics ; DNA-Binding Proteins ; genetics ; Fusion Proteins, bcr-abl ; genetics ; Genetic Predisposition to Disease ; Humans ; In Situ Hybridization, Fluorescence ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; pathology ; MDS1 and EVI1 Complex Locus Protein ; Male ; Neoplasm Proteins ; genetics ; Oncogene Proteins, Fusion ; genetics ; Proto-Oncogenes ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Transcription Factors ; genetics ; Translocation, Genetic

Objective To compare the antiarrhythmic effects and safety of landiolol and esmolol .Methods The sequential method was used to detect the median lethal dose ( LD50 ) and 50%effective dose ( ED50 ) of the two drugs.Treatment index were calculated by LD 50/ED50 .Several of arrhythmia models were used to study the antiarrhythmic effects of landio-lol and esmolol in the equivalent dose .Results ED50 and LD50 of landio-lol and esmolol were 14.4 , 17.3 mg? kg -1 and 347 , 100 mg? kg -1 re-spectively , TI were 24.1 and 5.8.The ratio of equivalent dose between landiolol and esmolol on anti -chloroform induced arrhythmia was 0.83∶1.Landiolol and esmolol have similar anti -arrythmia effect on va-rious arrhythmia models.Esmolol has a stronger inhibition function on the heart rate than that of landiolol .Conclusion Compared with esmolol , landiolol has similar antiarrhythmic effect , gentle and slow effect on heart rate, higher treatment index and safety in equivalent dose .

Objective To investigate the current situation in Chinese rheumatologic physicians' clinical diagnosis and evaluation of Takayasu's arteritis (TA).Methods Nineteen rheumatology experts and three vascular surgery specialists in China were invited to make the nationwide investigation for the first time about the diagnosis and disease activity evaluation of TA in China,through the questionnaire survey on the internet.Weighted average was used to calculate the average scores of corresponding problems.Results Chinese experts mainly adopted 1990 American College of Rheumatology (ACR) classification criteria for clinical diagnosis of TA.In details,symptoms of age,limb claudication and amaurosis,signs including pulselessness or pulse weakening,vascular bruits,increasing bilateral pulse pressure and hypertension and acute phase reactants (APR) were critical to the clinical diagnosis of TA.Besides,noninvasive imaging examinations,such as computed tomography angiography (CTA),magnetic resonance angiography (MRA),vascular ultrasonography,and positron emission tomography (PET) were also of great importance.In the aspect of disease activity assessment,Chinese experts mainly used Kerr scoring tool.APR and noninvasive radiological examinations were considered with vital value.Some TA patients with carotid artery involvement were recommended using vascular ultrasonography,while others with pulmonary artery and thoracic/abdominal aorta trunk involvement were preferred CTA other than MRA.Conclusions APR and noninvasive imaging examinations were thought with great help to make clinical diagnosis and evaluation of TA for Chinese physicians.