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 explore the specificity of anti-phosphotyrosine monoclonal antibody PY20 in bcr-abl+ cells and its possible clinical applications.

METHODSBcr-abl cell lines( K562, MEG-01) and bcr-abl- cells lines( Jurkat, MCF-7 )were stained with PY20. Phosphotyrosine protein of K562 and MEG-01 cells was detected by flow cytometry before and after treatment with imatinib. Phosphotyrosine protein in bone marrow cells from 49 patients with chronic myeloid leukemia (CML), Ph+ acute lymphoblastic leukemia(Ph(+) -ALL), Ph- ALL, acute myeloid leukemia (AML-M1, M2, M3, M5, FAB classification), chronic lymphocytic leukemia (CML) and 3 normal donor. Positive cells over 5% of total cells was considered positive cases for phosphotyrosine protein. The level of tyrosine phosphorylation was determined by median fluorescence intensity (MFI).

RESULTSBcr-abl cell lines and marrow cells from 10 CML patients and 8 ALL patients were all PY20-positive, while bcr-abl- cell lines and marrow cells from 18 leukemia patients and 3 normal donor were all PY20-negative. MFI decreased remarkably after blocked by imatinib in K562 cells and MEG-01 cells. The positive cell percent of marrow cells from 10 newly diagnosed CML patients and 9 imatinib-sensitive CML patients was (54.20 +/- 19.82)% and (14.84 +/- 6.17)% (P < 0.05), while that of 2 cases of imatinib-resistant was 64.3% and 57.2%. There was significant difference of MFI between imatinib-resistant patients and imatinib-sensitive patients (99.42 +/- 4.87 vs 46.41 +/- 4.67, P < 0.01).

CONCLUSIONPY20 monoclonal antibody is highly specific for bcr-abl+ cells. It might be useful in rapid detection of bcr-abl+ cells and sensitivity to imatinib of CML patients.

Antibodies, Monoclonal ; analysis ; Bone Marrow Cells ; metabolism ; Flow Cytometry ; Fusion Proteins, bcr-abl ; analysis ; Humans ; K562 Cells ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; metabolism ; Leukemia, Myeloid, Acute ; metabolism ; Phosphotyrosine ; analysis ; immunology

OBJECTIVETo examine the role of Polo-like kinase 1(PLK1) in the migration and invasiveness of human colorectal cancer cells.

METHODSNine colorectal cancer cell lines were cultured. Cell lines with the highest level of PLK1 expression was selected by PCR and Western blot. Three siRNA oligo segments targeting PLK1 were designed and selected cell lines transfected. Successful transfection was confirmed using real-time PCR and Western blot. Changes in migration and invasiveness of the selected cell line were evaluated by Transwell test.

RESULTSColorectal cancer cell line SW1116 was selected with the highest expression of PLK1 at both mRNA level and protein level. The expression of PLK1 in SW1116 was reduced by the three siRNA oligo segments to varying degrees, and the No.1 siRNA oligo segment was the most efficient. In migration test, the number of cells crossing through chambers in PLK1-siRNA group was 44 ± 14, which was lower than that in the negative control group (242 ± 40) and in blank control group(240 ± 38). In invasion test, the number of cells crossing through chambers in PLK1-siRNA group was 62 ± 3, which was lower than that in negative control group (207 ± 12) and in blank control group (211 ± 15). These differences were statistically significant(P<0.01).

CONCLUSIONPLK1 silencing by siRNA may inhibit the migration and invasiveness of colorectal cancer cells, suggesting that PLK1 might play an important role in the infiltration and metastasis of colorectal cancer.

Cell Cycle Proteins ; genetics ; metabolism ; Cell Line, Tumor ; Cell Movement ; Colorectal Neoplasms ; metabolism ; pathology ; Humans ; Neoplasm Invasiveness ; Protein-Serine-Threonine Kinases ; genetics ; metabolism ; Proto-Oncogene Proteins ; genetics ; metabolism ; RNA, Small Interfering ; genetics ; Transfection

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

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

OBJECTIVETo determine the influence of hand-sewn and stapler in distal pancreatectomy on the postoperative complication.

METHODSClinical data of 109 patients after distal pancreatectomy from January 2003 to December 2006 were analyzed retrospectively.

RESULTSThe surgical techniques used for closure of the pancreatic stump after distal pancreatectomy were categorized into hand-sewn closure group (n = 53) and stapler closure group (n = 56). In stapler closure group, 25 patients accepted laparoscopic operation. The incidences of abdominal infection and pancreatic fistulae in stapler closure group were lower than hand-sewn closure group. The operation time, blood infusion, postoperative bleeding and medical costs were similar between two groups.

CONCLUSIONStapler closure in distal pancreatectomy could decrease the incidence of pancreatic fistula and abdominal infections.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Male ; Middle Aged ; Pancreatectomy ; adverse effects ; instrumentation ; methods ; Pancreatic Fistula ; etiology ; prevention & control ; Pancreatic Neoplasms ; surgery ; Postoperative Complications ; prevention & control ; Retrospective Studies ; Surgical Staplers ; Surgical Wound Infection ; prevention & control ; Young Adult

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 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