Objective To investigate on influence of CYP2D6 genetic polymorphism on pharmacokinetics of tramadol in Chinese volunteers.Methods Adult healthy Chinese volunteers with different CYP2D6 genotypes were categorized into the following four groups: group 1:CYP2D6 * 2W * 10W, group 2:CYP2D6 * 2M * 10W, group 3:CYP2D6 * 2M * 10H, group 4 : CYP2D6 * 2M * 10M. After oral ad-ministration of 100 mg tramadol, plasma and urine samples were col-lected from each subject at different time within 32 h. The plasma and urine concentrations of tramadol and its metabolite O - desmethyltramad-ol (M1) were determined by HPLC with fluorescence detection. Re-suits The main pharmacokinetic parameters of tramadol and M, in group 2 were not significantly different from those in group 1. There are significant difference for the main pharmacokinetic parameters of tram-adol and M1 between group 3 and group 1, group 4 and group 1, group 4 and group 3, respectively (P<0.05 ). Conclusion The present re-sults shown that CYP2D6 * 2 has no influence on the pharmacokinetics of tramadol, but CYP2D6 * 10 reduces CYP2D6 activity which leads to the change of phenotype, and the homozygotes has more significant in-fluence on the pharmacokinetics of tramadol than the heterozygotes in Chinese population.

This study aimed to assess the relationship of OAS2 rs739901 5'-flanking C/A polymorphisms with the susceptibility to Enterovirus-71 (EV71) infection.We investigated 294 hand-foot-mouth disease (HFMD) Chinese children with EV71 infection (165 mild cases and 129 encephalitis cases).The improved multiplex ligation detection reaction (iMLDR) technique was used to test the genotypes.In EV71-infected patients,the CA genotype distribution (P=0.007),A allele frequency (OR 1.32,95％ CI 1.0-1.7,P=0.034)and CA+AA carriage frequency (P=0.003) of OAS2 rs739901 5'-flanking were obviously elevated as compared with controls,but there were no statistically significant differences between mild cases and encephalitis cases.In EV71-infected patients,the counts of white blood cells (P=0.034) and blood glucose concentrations (P=0.042) were raised in A carriers (CA+AA).Among different genotypes of encephalitis cases,the contents of cerebrospinal fluid (CSF) showed no significant differences.IFN-γ levels in EV71-infected patients were higher than those in controls (mild group vs.control group,P＜0.01;encephalitis group vs.control group,P＜0.001).In encephalitis cases,IFN-γ levels were reduced (P＜0.05) in A carriers compared to CC genotype,however,there were no significant differences between genotypes CA and AA (P=0.226).These findings suggest that OAS2 rs739901 5'-flanking C/A genetic polymorphisms involve the susceptibility to EV71 infection,and A allele might be a risk factor of the susceptibility to EV-71 infection.

Objectives:To understand the current situation of the sense of uselessness of the elderly in China and the impact of Internet use on the sense of uselessness of the elderly,exploring the moderating role of age and social participation in it,so as to provide empirical evidence for improving the mental health of the elderly and promoting active aging.Method:T-test and analysis of variance were used for univariate analysis,multiple linear regression was used for multivariate analysis,and point selection and Johnson Neyman methods were used to analyze the moderating effects of age and social participation.Results:At present,the level of the sense of uselessness of the elderly in China is relatively high.Internet use affects the sense of uselessness of the elderly(b=-0.10,P<0.001);With the increase of age and the level of social participation,the alleviating effect of Internet use on the sense of uselessness of the elderly has gradually weakened(b=0.01,P<0.05;b=0.02,P<0.01).Internet use can alleviate the sense of uselessness of the elderly and is regulated by age and social participation.It is suggested that the Internet access intervention should be carried out as soon as possible for the elderly at a lower age,and the elderly with a lower level of social participation should be focused,so as to play the positive role of the Internet in reducing the sense of uselessness of the elderly and improving their mental health.

OBJECTIVETo investigate the relationship between three types of bcr/abl fusion transcripts and clinical manifestation in chronic myeloid leukemia (CML).

METHODM-, m- and micro -bcr/abl fusion transcripts were detected by reverse transcriptase-polymerase chain reaction (RT-PCR) technique in 537 fresh bone marrow samples of patients suspected CML clinically.

RESULTSOf 573 patients, 479 expressed M-bcr/abl transcripts, among whom 370 were in chronic phase (CP), and 109 in accelerated (AP)/blastic phase (BP). The percentages of patients with b2a2 transcripts in CP and AP/BP were 32.4% (120/370) and 36.7% (40/109) (P > 0.05). The b2a2 transcript patients in blastic crisis were 52.6% (10/19) for lymphoblastic and 33.3% (30/90) for myeloblastic (P > 0.05). The platelet count of untreated patients with b3a2 isoform [(485.9 +/- 333.8) x 10(9)/L, n = 125] was distinctly higher than those with b2a2 isoform [(380.5 +/- 321.9) x 10(9)/L, n = 62] (P < 0.05). 66.0% (31/47) and 64.4% (29/45) of the patients in CP and AP/BP respectively co-expressed M- and m-bcr/abl transcripts (P > 0.05). One patient expressed only m-bcr/abl transcript was of typical acute myeloblastic leukemia (AML). Both two micro -bcr/abl(+) patients were of typical CML.

CONCLUSIONSAlmost all typical CML patients express M-bcr/abl transcripts, most of them coexpress M-bcr/abl and m-bcr/abl transcripts, a few possesses only micro -bcr/abl fusion gene. m-bcr/abl(+) are usually associated with AML or CML in myeloblastic crisis besides acute lymphoblastic leukemia (ALL). Patients with b3a2 isoform are prone to higher platelet count before treatment.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; Female ; Fusion Proteins, bcr-abl ; genetics ; Genotype ; Humans ; Infant ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; Male ; Middle Aged ; Reverse Transcriptase Polymerase Chain Reaction ; Transcription Factors ; genetics

Expression of AML1/ETO mRNA was observed in bone marrow cells from 49 untreated leukemic patients, and continuously detected during different periods after chemotherapy (12 cases) or bone marrow transplantation (8 cases). The results showed that AML1/ETO mRNA could be expressed in cells from AML-M(2), AML-M(4) and MDS-RAEB-T patients. The positive expression changed into negative at different duration in patients who achieved complete remission either by chemotherapy (9 cases), allogeneic bone marrow transplantation (5 cases) and autologous peripheral blood stem cell transplantation (1 case), and they were sustained in complete remission status. In chemotherapeutic group, patients whose AML1/ETO expression turning from negative (2 cases) or faint positive (1 case) to positive relapsed later. Two patients treated with Allo-BMT showed continuously positive results and died of GVHD and relapse, respectively. These observations suggest that AML1/ETO chimeric mRNA could disappeared after chemotherapy or bone marrow transplantation. The patients have a great probability to relapse if the results of RT-PCR are continuously positive or change from negative to positive. Regular detection is necessary for leukemic patients.

In order to investigate the features of M-bcr/abl and m-bcr/abl fusion transcripts in patients with chronic myeloid leukemia (CML) after allogeneic stem cell transplantation (SCT), M-bcr/abl and m-bcr/abl fusion transcripts were sequentially detected by RT-PCR technique in 72 CML patients after SCT. The results showed that M-bcr/abl positive rate (79.2%, 42/53) within 6 months after SCT was remarkably higher than that in 6-12 months group (34.3%, 11/32) and >or= 12 months group (35.1%, 13/37) (P < 0.001), and the clinical relapse rates in corresponding periods were 1.9% (1/53), 0% (0/32) and 16.2% (6/37) respectively. M-bcr/abl and m-bcr/abl fusion transcripts occurred in 5 of 6 clinically relapsed patients. In period of more than 6 months after transplantation, none of 17 M-bcr/abl(+) samples from 14 patients in cytogenetic remission appeared positive reaction of m-bcr/abl. It is concluded that M-bcr/abl(+) fusion transcript still existed in most patients after SCT, and usually disappeared within 6 months. Existence of M-bcr/abl is not a clinical relapse marker in CML patients. Simultaneous detection of M-bcr/abl and m-bcr/abl fusion transcripts can be helpful for monitoring residual disease.
Adolescent ; Adult ; Child ; Child, Preschool ; Female ; Follow-Up Studies ; Fusion Proteins, bcr-abl ; genetics ; Hematopoietic Stem Cell Transplantation ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; metabolism ; therapy ; Male ; Middle Aged ; RNA, Messenger ; analysis ; Recurrence ; Reverse Transcriptase Polymerase Chain Reaction ; Transplantation, Homologous

OBJECTIVETo investigate the unusual bcr/abl fusion gene structures of two Ph chromosome positive chronic myelogenous leukemia (CML) patients in chronic phase (CP).

METHODSBy using general M- and micro -bcr/abl specific primers respectively, bcr/abl fusion transcripts were detected by reverse transcription-polymerase chain reaction (RT-PCR). The RT-PCR products sequencing was performed, the DNA sequences were analyzed in Genebank and the bcr and abl sequences at the fusion site were identified. DNA was amplified by PCR using a set of primers designed according to the sequencing result of RT-PCR products.

RESULTSTwo patients showed typical manifestations of CML-CP. Their RT-PCR products were different from usual M- or micro -type; one was longer than M-bcr/abl but shorter than micro -bcr/abl, the other one was shorter than M-bcr/abl. The RT-PCR products sequencing showed that both products contained bcr and abl gene sequences. The first patient's bcr gene was broken within exon 18, and fused to abl gene exon 2(a2), and a 40 bp of partial abl intron 1b fragment was inserted between them, resulting in a novel in-frame bcr/abl fusion transcript-e18-int-a2 which has not been reported in the literature so far. In the second patient, deletion of abl exon2(a2) led to exon 13(b2) of bcr gene fusing with abl exon 3(a3).

CONCLUSIONUncommon bcr/abl fusion gene may occur in typical Ph(+) CML patient.

Adult ; Fusion Proteins, bcr-abl ; genetics ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; Male ; Molecular Sequence Data ; Philadelphia Chromosome ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA

In order to improve the fermentation potency of phytase in recombinant host and decrease the production cost, the pichia expression vector pGAPZalpha-A was modified by introduction of an AOX1 promoter from vector pPIC9 and the resulted vector pAOXZalpha is an methanol induced vector. After that, a phytase gene appA-m was cloned into pAOXZalpha to construct the recombinant vector pAOXZalpha-appA-m. The recombinant Pichia pastoris 74#, which already contains one copy of appA-m and its fermentation potency exceeded 7.5 x 10(6) IU/mL, was used as the host strain for the transformation of pAOXZalpha-appA-m. The Pichia pastoris transformants were gained by electroporation. PCR results indicated that the appA-m expression box has integrated into the genome of Pichia pastoris and the original construction of phytase gene has not changed. SDS-PAGE analysis revealed that phytase was overexpressed and secreted into the medium supernatant. Recombinants with high expression level were screened and used for fermentation. In 5L fermentor, the expression level of phytase protein achieved 4 mg/mL and the phytase activity (fermentation potency) exceeded 1.2 x 10(7) IU/mL, which was about 1.6-fold compared with that of the host strain 74#. Moreover, the improved recombinant Pichia pastoris is excellent at expression stability and heredity stability.
6-Phytase ; genetics ; Fermentation ; Gene Dosage ; Pichia ; genetics ; Plasmids ; Polymerase Chain Reaction ; Recombination, Genetic

OBJECTIVETo study the risk factors of postoperative gastroparesis syndrome after pancreaticoduodenectomy.

METHODClinical data of 166 patients who underwent pancreaticoduodenectomy from January 2002 to April 2006 were analyzed with binary logistic regression in order to explore the risk factors of postoperative gastroparesis syndrome after pancreaticoduodenectomy.

RESULTSThere were 60 patients occurred postoperative gastroparesis syndrome after pancreaticoduodenectomy. Among 15 factors, 6 factors were proved to be the risk factors of postoperative gastroparesis syndrome after pancreaticoduodenectomy by unitary anylasis; 7 factors were proved related factors of postoperative gastroparesis syndrome after pancreaticoduodenectomy by analyzed with binary logistic regression.

CONCLUSIONSThe risk factors are preoperative gastro-intestinal obstruction syndrome, extensive lymphadenectomy, postoperative abdominal cavity infection, starting time of postoperative enteral nutrition, postoperative blood glucose level; The protective factors are preoperative percutaneous transhepatic cholangial drainage (PTCD) and postoperative blood terminal protein (TP) level.

Adolescent ; Adult ; Aged ; Gastroparesis ; epidemiology ; Humans ; Logistic Models ; Middle Aged ; Pancreaticoduodenectomy ; adverse effects ; Postoperative Complications ; epidemiology ; Retrospective Studies ; Risk Factors

BACKGROUNDTumstatin is a novel endogenous angiogenesis inhibitor which is widely studied using purified protein. The current study evaluates the antiangiogenic effects of tumstatin-overexpression plasmid in vitro, reveals the mechanism underlying the vascular endothelial cell growth inhibition and searches for a novel method administering tumstatin persistently.

METHODSThe eukaryotic expression plasmid pcDNA-tumstatin encoding tumstatin gene was constructed and transfected to human umbilical vein endothelial cell ECV304 and human renal carcinoma cell ACHN. Expression of tumstatin in the two cell lines was determined by RT-PCR and Western blotting. Vascular endothelial cell proliferation was assessed by CCK-8 assay and cell cycle was analyzed by flow cytometry. To investigate the mechanism by which pcDNA-tumstatin inhibited vascular endothelial cell proliferation in vitro, cyclin D1 protein was detected by Western blotting.

RESULTSDNA sequence confirmed that pcDNA-tumstatin was successfully constructed. RT-PCR and Western blotting indicated that tumstatin could express in the two cell lines effectively. After tumstatin gene transfer, ECV304 cell growth was significantly inhibited and the cell cycle was arrested in G1 phase. And Western blotting showed that pcDNA-tumstatin decreased the level of cyclin D1 protein.

CONCLUSIONSOverexpression of tumstatin mediated by pcDNA 3.1 (+) specially inhibited vascular endothelial cells by arresting vascular endothelial cell in G1 phase resulting from downregulation of cyclin D1 and administration of tumstatin using a gene therapy might be a novel strategy for cancer therapy.

Autoantigens ; genetics ; metabolism ; Blotting, Western ; Cell Cycle ; genetics ; physiology ; Cell Line ; Cell Line, Tumor ; Cell Proliferation ; Collagen Type IV ; genetics ; metabolism ; Endothelial Cells ; cytology ; metabolism ; Flow Cytometry ; Humans ; Plasmids ; genetics ; Reverse Transcriptase Polymerase Chain Reaction