Histiocytoma, Malignant Fibrous ; pathology ; Humans ; Laryngeal Neoplasms ; pathology ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; pathology ; Male ; Middle Aged ; Neck

Child, Preschool ; Diagnostic Errors ; Female ; Humans ; Nasopharyngeal Neoplasms ; diagnosis ; Rhabdomyosarcoma, Embryonal ; diagnosis

The preliminary metabolic profile of total diterpene acid (TDA) isolated from Pseudolarix kaempferi was investigated by using in vivo and in vitro tests. Pseudolaric acid C2 (PC2) was identified as the predominant metabolite in plasma, urine, bile and feces after both oral and intravenous administrations to rats using HPLC-UV and HPLC-ESI/MS(n), and demethoxydeacetoxypseudolaric acid B (DDPB), a metabolite proposed to be the glucoside of PC2 (PC2G), as well as pseudolaric acid C (PC), pseudolaric acid A (PA), pseudolaric acid A O-beta-D glucopyranoside (PAG), pseudolaric acid B O-beta-D glucopyranoside (PBG) and deacetylpseudolaric acid A (DPA) originated from TDA could also be detected. It was demonstrated by tests that the metabolism of TDA is independent of intestinal microflora, and neither of pepsin and trypsin is in charge of metabolism of TDA, TDA is also stable in both pH environments of gastric tract and intestinal tract. The metabolites of TDA in whole blood in vitro incubation were found to be PC2, DDPB and PC2G, which demonstrated that the metabolic reaction of TDA in vivo is mainly occurred in blood and contributed to be the hydrolysis of plasma esterase to ester bond, as well as the glucosylation reaction. These results clarified the metabolic pathway of TDA for the first time, which is of great significance to the in vivo active form and acting mechanism research of P. kaempferi.
Animals ; Chromatography, High Pressure Liquid ; Diterpenes ; metabolism ; Glucosides ; metabolism ; Hydrolysis ; Mass Spectrometry ; Metabolic Networks and Pathways ; Pinaceae ; chemistry ; Rats

OBJECTIVETo explore the efficacy and determine the risk factors of survival for recurrent hepatocellular carcinoma(HCC) treated by percutaneous radiofrequency ablation (PRFA).

METHODSFrom January 1999 to December 2008, 82 patients with recurrent HCC, with the diameter less than 7 cm for solitary tumor, or the largest tumor less than 5 cm for multiple tumors(the number of tumors less than 3), were treated by PRFA. The significance of 12 clinical or pathological variables in the risk factors of overall survival were assessed.

RESULTSThe overall survival 1-, 3-, and 5-year survival rates were 75.8%, 43.9% and 34.5% (from the date of PRFA), and 95.1%, 63.2% and 46.6% (from initial hepatectomy), respectively. Univariate analysis indicated that tumor size before initial hepatectomy, recurrence interval from initial hepatectomy, number of recurrent tumors, diameter of largest recurrent tumor, serum glutamyl transpeptidase (GGT) and serum albumin (ALB) level were significant prognostic factors (P < 0.05, Kaplan-Meier Log-rank test). Multivariate analysis showed recurrence interval from initial hepatectomy, diameter of largest recurrence tumor, serum GGT and ALB level were significant prognostic (P < 0.05).

CONCLUSIONPRFA is effective for recurrent HCC. Recurrence interval from initial hepatectomy, diameter of largest recurrent tumor, serum GGT and ALB level are significant prognostic factors.

Adult ; Carcinoma, Hepatocellular ; pathology ; surgery ; Catheter Ablation ; Female ; Follow-Up Studies ; Humans ; Kaplan-Meier Estimate ; Liver Neoplasms ; pathology ; surgery ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; surgery ; Prognosis ; Retrospective Studies ; Risk Factors

OBJECTIVETo investigate the association of -141C insert/delete polymorphism with schizophrenia in Wuhan of Hubei province.

METHODSA case-control study was conducted to analyze the polymorphism in the D(2) receptor gene promoter region with schizophrenia. A total of 120 cases of schizophrenia diagnosed according to CCMD-II R criteria and 100 normal controls were recruited in the study.

RESULTSIn this sample, the allele and genotype showed statistically significant differences between patients and normal controls (P<0.05).Especially, the frequency of -141C del was 11% in patients and 18% in control(OR 0.55, 95% CI 0.30-0.96; P<0.05). This allele was less common in schizophrenia than in normal controls (P<0.05).

CONCLUSIONThe -141C del polymorphism is associated with schizophrenia.The polymorphism may modify the association with other factors. Possibly -141C del in the DRD(2) promoter region is a strong candidate for a protective factor for this trait.

Adult ; Alleles ; Female ; Gene Frequency ; Genotype ; Humans ; Male ; Middle Aged ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Promoter Regions, Genetic ; genetics ; Receptors, Dopamine D2 ; genetics ; Schizophrenia ; diagnosis ; genetics

OBJECTIVE: To construct two types of Wnt-inducible secreted protein 3(WISP3) gene's mutants(1000T/C,840delT) found in spondyloepiphyseal dysplasia tarda with progressive anthopathy (SEDT-PA) patients, and to observe their expression in COS-7 cells. METHODS: Full-length cDNA of wild type WISP3 gene(WT-WISP3) was amplified from human chondrocytes by RT-PCR, and site-directed mutagenesis was used to obtain full-length cDNAs of the mutated WISP3 genes(MUT1000T/C and MUT840delT). The recombined plasmids WT-WISP3/pcDNA3.1(+), MUT1000T/C/pcDNA3.1(+) and MUT840delT/pcDNA3.1(+) were transfected transiently into COS-7 cells by liposome-mediated method, and pcDNA3.1(+) vector was used as a control. The total RNA and protein of the transfected COS-7 cells were extracted after 48 hours of transfection. The expression of WISP3 gene in the transfected COS-7 cells was detected by semi-quantitative RT-PCR and Western blot. RESULTS: By restriction endonuclease analysis and sequencing, the sequence of MUT1000T/C and MUT840delT were consistent with that mutated in SEDT-PA, and the open reading frames matched with the vector sequence. Semi-quantitative RT-PCR and Western blot showed that the recombined plasmids were highly expressed in COS-7 cells. CONCLUSION WISP3 gene's mutants of SEDT-PA are successfully constructed by genetic recombination, and expressed in COS-7 cells, which lays the foundation for the further study on its molecular functions in SEDT-PA.
Animals ; Base Sequence ; CCN Intercellular Signaling Proteins ; COS Cells ; metabolism ; Chlorocebus aethiops ; Humans ; Insulin-Like Growth Factor Binding Proteins ; biosynthesis ; genetics ; Molecular Sequence Data ; Mutagenesis, Site-Directed ; Mutation ; Osteochondrodysplasias ; genetics ; metabolism ; Transfection

Catheter Ablation ; Child ; Heart Septal Defects, Atrial ; surgery ; Humans ; Imaging, Three-Dimensional ; Male ; Tachycardia, Supraventricular ; surgery ; Tomography, X-Ray Computed

OBJECTIVETo observe early rehabilitation of two different acupuncture programs for hemiplegia due to stroke.

METHODSSeventy cases of early hemiplegia due to stroke were randomly divided into a treatment group and a control group. The treatment group were treated by acupuncture at acupoints of both the healthy and affected sides, twice each day, respectively, and the control group by acupuncture at the affected side, once daily. Their therapeutic effects were evaluated by the brief Fugl-Meyer movement scale and the modified Barthel indexes.

RESULTSThe patients with over grade IV for FMA accounted for 68.6% and the good rate for ADL was 74.3% in the treatment group, and 31.4% and 42.8% in the control group, respectively, the therapeutic effect of the treatment group being better than that of the control group (P<0.01 or P<0.05).

CONCLUSIONThe acupuncture program in which acupoints at both the healthy and affected sides are applied alternately is more beneficial to recovery of motor function and activity daily living (ADL) of the patient with hemiplegia due to stroke.

Activities of Daily Living ; Acupuncture Points ; Acupuncture Therapy ; Hemiplegia ; therapy ; Humans ; Stroke ; therapy

OBJECTIVETo investigate the synergistical killing effect of docetaxel combined with ABT-737 on human prostate cancer cell line PC-3 by inducing apoptosis and further to determine the mechanism underlying such effect.

METHODSPC-3 cells were treated with various concentrations of docetaxel or (and) ABT-737. Cell viability was determined using MTT assay. Apoptosis was assessed by fluorescence microscopy analysis of cells with condensed and segmented nuclei following staining with 4',6-diamidino-2-phenylindole (DAPI). Cellular DNA was stained with propidium iodide and flow cytometric analysis was performed to analyze the cell cycle distribution. Bcl-2, Bax, Bcl-xL and Mcl-1 protein changes were detected by Western blot. The activity of caspase-3 was measured using a colorimetric assay.

RESULTSDocetaxel (20 nmol/L) combination with ABT-737 (400 nmol/L) for 48 hours, the cell viability was decreased to 19.7% ± 3.2% to compare with 44.2% ± 4.4% (t = 4.45) of docetaxel and 93.2% ± 1.8% of ABT-737 separately and there was a synergistic effect between the two drugs (CI = 0.8). Apoptosis rate of the combination group was higher than other two drugs. Docetaxel increased the cell number arrested in G(2)/M phase compared with control group (P < 0.05), but the combination treatment resulted in a significant arrest in the G(0)/G(1) phase. The combination treatment could significantly reduced the Bcl-2, Bcl-xL and Mcl-1 expression (F = 369.53, 57.89 and 32.77, all P < 0.05) and enhanced the activity of caspase-3 (419.7% ± 15.6%) (F = 207.33, P < 0.05).

CONCLUSIONSThe combination of ABT-737 with docetaxel can synergistically inhibit the proliferation of PC-3 cells through inducing apoptosis, which may be associated with cell cycle arrest, down-regulation of Bcl-2, Bcl-xL and Mcl-1 expression and activation of caspase-3.

Apoptosis ; drug effects ; Biphenyl Compounds ; pharmacology ; Caspase 3 ; metabolism ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Drug Synergism ; Humans ; Male ; Myeloid Cell Leukemia Sequence 1 Protein ; metabolism ; Nitrophenols ; pharmacology ; Piperazines ; pharmacology ; Prostatic Neoplasms ; metabolism ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Sulfonamides ; pharmacology ; Taxoids ; pharmacology ; bcl-X Protein ; metabolism

Carboxypeptidase B is a metalloenzyme, which is widely used for commercial and research purposes. Commercially available CPB purified from porcine or bovine pancreas is very expensive, and is not totally free from other proteases. In order to express the rat proCPB in Pichia pastoris, total RNA extracted from SD rat pancreas cells was reversely transcripted to synthesize cDNA, and the proCPB ORF was synthesized by PCR. After digestion with Xho I and EcoR I , the fragment was inserted into pPIC9, and the recombinant plasmid was named as pPIC9-proCPB. By digestion with Sac I , the lined pPIC9-proCPB was transformed into Pichia pastoris strains GS115 with PEG1000 and integrated into their genomes. In the inducement of methanol, recombinant proCPB was successfully expressed in Pichia pastoris, and could be secreted into the supernatant in the culture. After optimizing the fermentation conditions, a higher production could be obtained when GS115-proCPB was induced in BMGY (pH6.0) at 28CC, with addition of 0.5% casein. The yield of recombinant protein reached 500mg/L, achieving over 94% of total protein in the culture supernatant. The purity of recombinant CPB can reach 96% after two step phenyl sepharose F F purification, and 38% of total protein can obtained after optimizing the pufication method. Comparing to the specific activity 180u/mg of CPB purchased from Sigma, the specific activity of recombinant CPB is 110u/mg. Mass spectrometry analyses showed the mass of the recombinant CPB was 35.1 kD, which is very close to the theory value 35.2 kD. Amino acid sequencing of N-terminal of recombinant CPB further indicated proCPB was expressed successfully and modificated correctly after translation.
Animals ; Carboxypeptidase B ; genetics ; isolation & purification ; metabolism ; Catalysis ; Cattle ; Electrophoresis, Polyacrylamide Gel ; Gene Expression Regulation, Enzymologic ; Kinetics ; Mass Spectrometry ; Molecular Weight ; Pichia ; genetics ; Rats ; Rats, Sprague-Dawley ; Recombinant Proteins ; chemistry ; isolation & purification ; metabolism ; Sequence Analysis, Protein ; Substrate Specificity ; Swine