OBJECTIVETo investigate the effect of extracellular regulating kinase (ERK) signaling pathway on the secretion of gamma-aminobutyric acid (GABA) in cultured rat hippocampal neurons induced by stromal cell derived factor-1 (SDF-1).

METHODSThe hippocampal neurons of newborn SD rats were cultured and identified in vitro; the phosphorylation level of ERK1/2 was examined by Western blot; ELISA was used to detect the effect of PD98059, a ERK1/2 specific blocker on GABA secretion of cultured hippocampal neurons and Western blot were adopted to measure the protein expression levels of glutamate decarboxylase (GAD65/67) and gamma aminobutyric acid transporter (GAT); after blocking ERK1/2 signaling pathway with PD98059; RT-PCR was used to detect the mRNA expression levels of GAT-1 and GAD65 after treated with PD98059.

RESULTSThe levels of ERKl/2 phosphorylation were increased significantly by SDF1 acting on hippocampal neurons, and CX-CR4 receptor blocker AMD3100, could inhibit SDF-1 induced ERK1/2 activation; SDF-1 could inhibit the secretion of GABA in cultured hippocampal neurons, and ERK1/2 specific inhibitor PD98059, could partly reverse the inhibition of GABA secretion by SDF-1. The effects of SDF-1 on cultured hippocampal neurons was to decrease the mRNA genesis of glutamic acid decarboxylase GAD65 and GABA transporter GAT-1, besides, ERK inhibitor PD98059 could effectively flip the effect of SDF-1. The results of Western blot showed that SDF-1 could inhibit the protein expression of GAT-1 and GAD65/67 in hippocampal neurons and the inhibition of GAT-1 and GAD65/67 protein expression could be partially restored by ERK1/2 blocker.

CONCLUSIONSDF-1 acts on the CXCR4 of hippocampal neurons in vitro, and inhibits the expression of GAD by activating the ERK1/2 signaling pathway, and this may represent one possible pathway of GABA secretion inhibition.

Animals ; Blotting, Western ; Cells, Cultured ; Chemokine CXCL12 ; pharmacology ; Flavonoids ; pharmacology ; Glutamate Decarboxylase ; metabolism ; Hippocampus ; cytology ; MAP Kinase Signaling System ; Neurons ; metabolism ; Phosphorylation ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Receptors, CXCR4 ; metabolism ; gamma-Aminobutyric Acid ; secretion

Hot Temperature ; Humans ; Occupational Exposure ; Occupational Medicine ; standards

Objective: To investigate the role of human umbilical vein endothelial cells (HUVEC) in supporting hematopoiesis and the expansion of hematopoietic stem/progenitor cells in vitro. Methods: According to the fact that HUVEC supernatant has colony stimulating activity shown by methylcellulose colony-forming assay and HUVEC can maintain the survival of mononuclear cells for at least four weeks in vitro, CD34+ cells from umbilical cord blood were seeded with (HUVEC group) or without (control group) HUVEC monolayer. Every week cells were collected and counted, the frequency of CFU-GM was measured by using methylcellulose colony-forming assay, and the percentage of CD34+ and CD41a+ cells was measured by flow cytometry. Results: In control group，all the CD34+ cells died in two weeks. However, in HUVEC group,most nucleated cells and CD34+ cells were expanded by 68.1±14.8 fold and 6.6±1.4 fold，respectively at the third week while CFU-GM expansion reached its peak (5.7±2.1 fold) at the week 2. Moreover, the percentage of CD41a+ cells was enhanced significantly, reaching a maximum (15.6%) at the week 3. Conclusions:HUVEC can support hematopoiesis in vitro and expand the hematopoietic progenitor cells and CD41a+ cells in direct contact coculture.

Literatures on pain intervention with transcutaneous electrical acupoint stimulation (TEAS) were collected by searching the databases both in Chinese and English, and summarized to understand the research progress of TEAS effects on pain mediators in recent years. This will provide a more objective and scientific theoretical basis for clinical practice of TEAS to treat pain syndrome, thus promoting the clinical application of TEAS. Our literature analysis indicated that TEAS effectively regulated the release levels of various pain factors such as prostaglandin, 5-hydroxytryptamine, interleukins, substance P and tumor necrosis factor-α to achieve the analgesic effects by affecting the conduction pathways. TEAS is a safe, non-invasive and effective treatment for pain syndrome. However, further research is necessary due to the lack of rigor of the current clinical trial design.

Objective To explore the effect and indication of splenectomy in liver transplantation.Methods From January 2001 to April 2006,260 patients underwent piggyback orthotopic liver transplantation(PBOLT),and 28 patients had undergone combined PBOLT and splenectomy(splenectomy group).These patients were compared to 56 randomly selected non-splenectomy patients from the same transplant period,meaningly two controls were se- lected for every non-spleneetomy case.Two groups were analyzed with respect to rate of infection and survival rate, as well as biopsy-proven acute allograft rejection within 30 days after transplantation.Results Rate of infection in the splenectomy group was higher than that in the non-splenectomy patients(85.7% vs 55.4%,P

OBJECTIVETo understand the epidemics and antigenic drift of influenza viruses in China from 2000 to 2001.

METHODSThe viruses were grown in embryonated hen eggs with 9 - 10 days old. The egg allantoic fluids with influenza viruses were used. Virion RNA was transcribed into cDNA by reverse transcriptase while cDNA amplified by PCR. Products of PCR were purified. RNA sequence analysis was then performed. Finally, phylogenetic analysis of the sequencing data was performed with MegAlign (Version 1.03) and Editseq (Version 3.69) software.

RESULTSData from comparison of amino acid sequence on HA1 domain of HA protein molecule between H1N1 viruses isolated in 2001 and A/Shanghai/7/99 (H1N1) strain indicated that there was only one difference of amino acid located at 190 position (antigenic determinant D). However, phylogenetic analysis showed that there were two distinguishable genetically lineages of H1N1 viruses co-circulating in men in China in 2001. Two antigenically distinct genetic lineages of influenza B viruses were still existing in men in China. Most of influenza B viruses were Yamagata-like strain and there were two different amino acid sequences located at 197 and 199 position on HA1 domain of HA protein molecule, between Victoria-like virus isolated and B/Shandong/7/97 strain. When comparing amino acid sequences on HA1 protein domain of H3N2 viruses isolated in 2000 with those of A/Sydeney/5/97 (H3N2) virus, it was revealed that there were 7 - 8 differences of amino acid sequences between them. However, there were four differences related to amino acid sequences on HA1 protein domain between H3N2 viruses isolated in 2000 and in 2001. These results were further demonstrated by analysis of phylogenic tree.

CONCLUSIONSInfluenza was not prevalent in China from 2000 to 2001. The antigenic drifts of H3N2 and B/Victoria-like viruses occurred. Two antigenically distinct genetic lineages of influenza B viruses were still co-circulating in men in China. Two genetically distinct lineages of influenza A (H1N1) virus were also co-circulating in men in China.

Antigens, Viral ; genetics ; China ; epidemiology ; DNA, Viral ; genetics ; Female ; Genes, Viral ; genetics ; Genetic Variation ; Hemagglutinin Glycoproteins, Influenza Virus ; genetics ; Humans ; Influenza A virus ; classification ; genetics ; immunology ; isolation & purification ; Influenza B virus ; classification ; genetics ; immunology ; isolation & purification ; Influenza, Human ; epidemiology ; virology ; Male ; Orthomyxoviridae ; classification ; genetics ; Phylogeny ; Reverse Transcriptase Polymerase Chain Reaction ; Sentinel Surveillance ; Sequence Analysis, RNA

BACKGROUNDTo understand the antigenic and genetic characteristics of Victoria like strain of influenza B virus isolated recently and to provide a scientific evidence for influenza surveillance and monitoring of influenza epidemic in future.

METHODSViruses were passed in embryonated hen eggs and virion RNA was extracted from allantoic fluid and reverse transcribed to synthesize cDNA. cDNA was amplified by PCR and the PCR product was purified with a purification kit. Afterwards RNA sequence analysis was performed by dideoxynucleotide chain termination and a cloning method. Finally, phylogenetic analysis of the sequencing data was performed with MegAlign.

RESULTSB/Sichuan/63/2001 and B/Zhejiang/2/2001 viruses were antigenically different from B/Shandong/7/97 strain. The substitution of nucleotide sequences of HA1genes of them compared with those of B/Shandong/7/97strain resulted in the change of amino acid sequences in antigenic determinants on HA1 protein domain. The phylogenetic analysis also indicated that strains isolated recently were genetically different from B/Shandong/7/97/strain. However, there was neither differences on the antigenicity nor genetic partern between these two isotates.

CONCLUSIONSThe antigenic drift of Victoria-like strain of influenza B virus isolated recently in China has further occurred.

Amino Acid Sequence ; Antigenic Variation ; China ; Epitopes ; Genetic Variation ; Hemagglutinin Glycoproteins, Influenza Virus ; genetics ; immunology ; Humans ; Influenza B virus ; genetics ; immunology ; Influenza, Human ; virology ; Molecular Sequence Data ; RNA, Viral ; analysis ; Sequence Analysis, RNA

OBJECTIVETo investigate the dynamic correlation between pre-S1 antigen, pre-S2 antigen and HBV DNA in the serum of chronic hepatitis B (CHB) patients undergoing nucleoside analogue therapy.

METHODS12 CHB patients with transient virological response after lamivudine treatment, and 20 patients treated with adefovir for 5 years were recruited in this study. Serum samples were collected at four time points when HBV DNA fluctuated sharply during lamivudine treatment, and at 0, 8, 12, 28, 52, 104, 156, 208, 260 weeks following adefovir treatment. HBV DNA was quantified by real-time PCR, pre-S1 and pre-S2 antigens were detected by ELISA.

RESULTSThe titers of pre-S1 and pre-S2 antigens were not correlated with the HBV DNA level in the serum of lamivudine treated patients. Only in one case of the adfovir treated patients, the decrease of pre-S1 and pre-S2 antigens was in parallel with the decrease of HBV DNA. Linear regression analysis indicated that neither pre-S1 antigen nor pre-S2 antigen was correlated with HBV DNA in the serum of lamivudine or adfovir treated patients (P more than 0.05).

CONCLUSIONOur results indicate that the titers of pre-S1 and pre-S2 antigens are not correlated with the serum HBV DNA in CHB patients undergoing nucleoside analogue therapy. Neither pre-S1 nor pre-S2 is a good predictor for the outcome of nucleoside analogue treatment.

DNA, Viral ; blood ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; drug therapy ; Humans ; Lamivudine ; therapeutic use ; Real-Time Polymerase Chain Reaction

OBJECTIVETo characterize DNA-PKcs and Ku70 expressions in hepato- and cholangio-neoplastic tissues and the association with the degree of malignancy and invasiveness of the tumors.

METHODSThe expression of DNA-PKcs and Ku70 was examined in 47 cases of hepato- or cholangio-neoplasm by immunohistochemistry.

RESULTSKu70 was expressed in all of the neoplastic tissues examined and with a little variation in levels. The highest expression was observed in adenocarcinomas and adenomas. There was no statistically significant association between Ku70 expression level and the degree of their malignancy extent or invasiveness. In contrast to Ku 70, a wide variation in expression levels of DNA-Pkcs was observed among different types of neoplastic tissues. The highest ratio of positive expressing cells was detected in hepatocellular carcinomas (92.1%), which was significantly higher than that in cholangioadeno carcinomas (65.3%) and biliary cystadenocarcinomas (51.9%). Low or no expression level was detected in papillary adenoma cases. DNA-PKcs expression of invasive adenomas and adeno-carcinomas (61.2%) was significantly higher than that of non-invasive adenomas and adeno-carcinomas (30.4%). There was no expression observed in the normal tissues adjacent to the tumors.

CONCLUSIONDNA-PKcs is expressed in hepato- and cholangio-neoplasms and its variable level of expression is associated with the types of the tumor and their degree of malignancy and invasiveness. DNA-PKcs could be recognized as a new biomarker for liver neoplasm.

Adenocarcinoma ; enzymology ; Adolescent ; Adult ; Aged ; Aged, 80 and over ; Antigens, Nuclear ; biosynthesis ; genetics ; Bile Duct Neoplasms ; enzymology ; Bile Ducts, Intrahepatic ; enzymology ; Biomarkers, Tumor ; biosynthesis ; genetics ; Carcinoma, Hepatocellular ; enzymology ; DNA-Activated Protein Kinase ; biosynthesis ; genetics ; DNA-Binding Proteins ; biosynthesis ; genetics ; Female ; Humans ; Ku Autoantigen ; Liver Neoplasms ; enzymology ; Male ; Middle Aged

OBJECTIVETo investigate the polymorphism of Kell blood group system in Chinese and to find a suitable method for large scale screening.

METHODSAn analysis method of polymerase chain reaction-restriction fragment-single strand conformation polymorphism (PCR-RF-SSCP) combined with heteroduplex was established to detect abnormal sample in KEL exon 7-9 area, then sequencing was used to find out the mutation site.

RESULTSTwo mutations were found from 500 samples: 966G > A mutation in exon 9 and C > A mutation in 67th site of intron 7, both with no amino acid change. The mutation rate was 4/1000. No mutation was found as missed in using PCR-RF-SSCP combined with heteroduplex.

CONCLUSIONPCR-RF-SSCP combined with heteroduplex is confirmed as an effective, economical and simple method, it is quite suitable for large scale population screening study with unclear gene background and unavailable positive controls. Since there is special polymorphism for Kell blood group system in Chinese, further study is needed.

Asian Continental Ancestry Group ; genetics ; China ; Heteroduplex Analysis ; methods ; Humans ; Kell Blood-Group System ; genetics ; Polymerase Chain Reaction ; methods ; Polymorphism, Restriction Fragment Length ; Polymorphism, Single-Stranded Conformational