OBJECTIVETo study the role of integrin alpha 6 in cell attachment, spread, survival/proliferation and differentiation of human hepatocellular carcinoma (HCC) BEL-7402 cells on various substrates by monoclonal antibody against the extracellular domain of alpha 6 subunit (IA6ED McAb).

METHODSThe effect of McAb on attachment and spread of BEL-7402 cells on LN or FN substrate was examined. MTT analysis was used to examine the cell survival/proliferation, gelatin zymography to the matrix metalloproteinases (MMPs) secreted by BEL-7402 cells, and microparticle immunoabsorbent kit to AFP secretion while the cells were cultured on LN-, FN- or Matrigel-coated substrates.

RESULTSThe cell attachment, spread and survival/proliferation were inhibited. Moreover importantly, the malignant cell dedifferentiation and abnormal differentiation on LN-coated substrate were also strongly inhibited by IA6ED McAb.

CONCLUSIONLN and integrin alpha 6 regulate human HCC cell phenotypes of survival/proliferation and differentiation. The phenotypes of dedifferentiation and abnormal differentiation can be reversed by blocking the interaction between integrin alpha 6 and LN using IA6ED McAb, which may lower metastatic potency of tumor cells.

Antibodies, Monoclonal ; pharmacology ; Carcinoma, Hepatocellular ; pathology ; Cell Adhesion ; drug effects ; Humans ; Integrin alpha6 ; immunology ; physiology ; Laminin ; physiology ; Liver Neoplasms ; pathology ; Phenotype ; Receptors, Laminin ; physiology ; Tumor Cells, Cultured

OBJECTIVETo evaluate the potential usefulness of a multivariable model, established mainly upon peripheral T-cell subsets, Th1/Th2, T-bet and GATA-3 gene expressions as well as TCM and Western medical diagnostic criteria, in predicting the selection of immunosuppressive therapy (IST) or androgens in treating patients with aplastic anemia (AA).

METHODSPeripheral blood T cell subsets in 85 patients with AA were serially analyzed by flow cytometry before and after treatment, and their T-bet and GATA-3 gene expressions were assessed meantime by Real-time PCR. Then analysis of Logistic regression equation and ROC curves were performed based on the cases responding to IST or androgens.

RESULTS(1) According to the logistic equation and ROC curve of SPSS, setting the false positive rate as 0.10, the P value was 0.832. When P> or =0.832, patients were judged in the immunosuppressive dominant state, IST should be applied; when P<0.832, it means patients in the bone marrow failure dominant state, androgens should be added. (2) A novel theory is raised by the above-mentioned analysis, which indicated that the genesis and development process of AA could be divided in 2 stages, the abnormal immune dominant stage and the bone marrow failure dominant stage. For treatment of patients in the two stages, IST and androgens is the preference respectively.

CONCLUSIONThe multivariable model could be used for indicating which stage the AA patient is in, the abnormal immune stage or the bone marrow failure stage, and thus to guide the proper selection of IST or androgens in clinical practice.

Adult ; Aged ; Androgens ; therapeutic use ; Anemia, Aplastic ; drug therapy ; immunology ; Cyclosporine ; therapeutic use ; Female ; GATA3 Transcription Factor ; metabolism ; Humans ; Immunosuppressive Agents ; therapeutic use ; Logistic Models ; Male ; Middle Aged ; T-Box Domain Proteins ; metabolism ; T-Lymphocyte Subsets ; immunology ; Young Adult

Objective To evaluate the effect of correcting upper eyelid retraction in patients with iatrogenic ophthalmopathy by a new technique of modified transsection and longitudinal suture levator muscle prolongation treatment.Methods The traditional levator muscle flap trapezoidal incision was changed into horizontal incision,According to the amount of upper eyelid retraction we designed a transverse incision with 1 ∶ 2 in the central part of levator aponeurosis.Along the line of incision was designed transverse incision of Mulller's muscle combined with levator aponeurosis complex was designed,with longitudinal suture of incision.Results Five patients (6 eyes) with iatrogenic upper eyelid retraction were treated by this method.The follow up was performed for 3 to 12 months.All were satisfied with good cosmetic results,the degree of bilateral corneal exposure was symmetrical,with no recurrence and complications,except for 1 cases of unilateral upper eyelid retraction width slightly asymmetry.Conclusions The application of modified levator muscle prolongation treatment is not only quite simple,easy to adjust in operation,but also with reliable effect and less complication.It is effective for iatrogenic upper eyelid retraction range from mild to moderate degrees of correction.

Objective To understand the prevalence and the factors associated with depression and anxiety among patients attending the community-based methadone maintenance treatment(MMT) program.Methods 9 MMT clinics in 3 provinces were randomly selected,in which 1 301 MMT patients who met the criteria of recruitment were all interviewed.A cross-sectional study by administrative questionnaire was employed to investigate the demographics,drug abuse and MMT treatment status of the respondents.The 1301 respondents were also evaluated for depression and anxiety by SDS(Self-Rating Depression Scale) and SAS(Self-Rating Anxiety Scale) respectively.ANOVA and multivariate linear regressions analysis was used to verify the factors associated with depression and anxiety respectively.Results The prevalence of depression and anxiety among the MMT patients were 38.3%(498/1 301) and 18.4%(239/1 301).The multivariates linear regression analysis indicated that at the level of ?=0.05,gender,employment status were associated with depression,while gender,employnment status,educational level,drug abuse and methadone daily dosage were associated with anxiety.Conclusions There are certain proportions of MMT clients who suffer from depression and anxiety.The MMT clinic staff should put more emphasis on the efforts to reduce psychological disorders and its potential risks by enhancing psychological councelling,intervention and social support towards the MMT patients.

This paper present a microcontroller system for target controlled infusion according to pharmacodynamic parameters of intravenous anesthetics. It can control the depth of anesthesia by adjusting the level of plasma concentrations. The system has the advantages of high precision, extended function and easy operation. It has been now used in the clinical anesthesia.
Algorithms ; Anesthesia, Intravenous ; instrumentation ; methods ; Anesthetics, Intravenous ; administration & dosage ; pharmacokinetics ; Computer Systems ; Humans ; Infusions, Intravenous ; Microcomputers ; Software Design

OBJECTIVE To explore the neuro-protective effects of saffron (Crocus satius L.) on chronic focal cerebral ischemia in rats.METHODS SD rats were randomly divided into 6 groups:sham control group,MCAO group,edaravone group and saffron 30,100,300 mg·kg-1groups.Focal cerebral ischemia was induced by middle cerebral artery occlusion(MCAO).Saffron was administered orally by once daily from 2 h to 42 d after ischemia. At 42 d after cerebral ischemia, neurological deficit score, spontaneous activity test,elevated plus maze test,marble burying test and novel objective recognition test were used to evaluate the effects of saffron on the behevioural change. Infarct volume, survival neuron density, activated astrocyte number, and the thickness of glial scar were also detected. GFAP expression and inflammatory cytokine contents in ischemic peripheral region were detected by Western blot and ELISA,separately.RESULTS Saffron(100,300 mg·kg-1)improved the body weight decrease, neurological deficit and spontaneous activity. Saffron (30-300 mg·kg- 1) increased the traveled distance ratio and total time in open arm, decreased the buried marble number, which indicated that saffron could ameliorate anxiety- and depression-like behaviors. Saffron (100, 300 mg·kg-1)improved the learning and memory function,which manifested by increased discrimination ratio(DR)and discrim-ination index (DI) in T2test. The results of toluidine blue found saffron treatment (100, 300 mg·kg-1) decreased the infarct volume and increased the neuron density in cortex and hippocampal.The activated astrocyte number,the thickness of glial scar and GFAP expression in ischemic peripheral region decreased after saffron. Saffron (100, 300 mg·kg-1) decreased the contents of IL-6 and IL-1β, increased the content of IL-10 in ischemic peripheral region.CONCLUSION Saffron exerted neuro-protective effects on chronic focal cerebral ischemia,which could be related with inhibiting the activation of astrocyte and glial scar,following with the decrease of inflammatory reaction.

We designed specific primers and fluorescence-labeled probes to develop real-time and conventional RT-PCR assays for detection of human coronavirus NL63 or HKU1. Subsequently, experiments were undertaken to assess diagnostic criteria such as specificity, sensitivity and reproducibility. The detection limit of the real-time RT-PCR assays was 10 RNA copies per reaction mixture. No cross-reactivity was observed between RNA samples derived from designed HCoV and other HCoV or human metapneumovirus. A total of 158 nasopharyngeal swab specimens collected from adult patients with acute respiratory tract infection in Beijing were screened for the presence of human coronavirus NL63 and HKU1 by using real-time RT-PCR and conventional RT-PCR method. The fluorescence quantitative RT-PCR method detected six specimens positive for human coronavirus NL63, five specimens positive for human coronavirus HKU1; and conventional RT-PCR method detected three HCoV-NL63 positive and three HCoV-HKU1 positive, respectively. The convention RT-PCR products of positive samples were obtained and sequence analysis confirmed the reliability of the above methods. In summary, the real-time RT-PCR assay for HCoV- NL63 or HKU1 was more sensitive than conventional RT-PCR and with less time (less than 4 hours) for completion. It may be suitable for molecular epidemiological surveillance and clinical diagnosis for human coronavirus NL63 and HKU1.
Coronavirus ; classification ; genetics ; isolation & purification ; Humans ; Nasopharynx ; virology ; Phylogeny ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Sensitivity and Specificity

Objective To isolate and study the phenotype and biological characteristics of chronic myelogenous leukemia-derived mesenchymal stem cells. Methods Cultivate the mesenchymal stem cells from chronic myelogenous leukemia in vitro, identify the immunophenotype by fluorescence-activated cell sorter (FACS), and detect the bcr-abl fusion gene by RT-PCR and FISH. Results The mesenchymal stem cells from chronic myelogenous leukemia can express the bcr-abl fusion gene. Conclusion These findings provide the evidence that the rearrangement of bcr-abl gene might happen at least more primitive than CD+34 CD-38 cells.

The spike (S) glycoprotein of HCoV-NL63 is a major target in the development of diagnostic assays and vaccines, but its antigenic and immunogenic properties remain unclear. Four fragments coding spike proteins (S1, S2, RL and RS) from HCoV-NL63 were amplified and cloned into the expression vector derived from vaccinia virus (Tiantan strain), and recombinant vaccinia viruses expressing four segments of spike proteins were generated (vJSC1175-S1; vJSC1175-S2; vJSC1175-RL; vJSC1175-RS), respectively. Their expression location in cell and level were characterized using indirect immune fluorescence assay (IFA) and Western-Blot, respectively. The expressions of four segments of spike proteins in recombinant vaccinia viruses were showed at appropriate level and with posttranslational modification (glycosylation), and S1, RL and RS were mainly distributed in the cell membrane, while the S2 was mainly distributed in the cytoplasm. Our results provide a basis for further exploring diagnostic role and vaccine development of different spike segments from HCoV-NL63.
Base Sequence ; Blotting, Western ; Coronavirus NL63, Human ; chemistry ; Fluorescent Antibody Technique, Indirect ; Humans ; Membrane Glycoproteins ; biosynthesis ; genetics ; Molecular Sequence Data ; Plasmids ; Recombinant Proteins ; biosynthesis ; Spike Glycoprotein, Coronavirus ; Vaccinia virus ; genetics ; Viral Envelope Proteins ; biosynthesis ; genetics

To investigate the genetic stability (including the vector of vaccinia virus and six foreign genes: gp160, gag, pol, rev, tat and nef) of the HIV-1 non-replicating recombinant vaccinia virus (rNTV-C). rNTV-C was serially passaged to passage 25 (P25) in primary chicken embryo fibroblast (CEF). P9, P12, P15 and P25 were selected to study the genetic stability in four aspects, including the genetic stability of viral vector, the genetic stability of six foreign genes, the expressing stability of foreign genes and the genetic loss of foreign genes. The results showed that the viral vector was non-replicated vaccinia virus of Tiantan strain and was passaged stably; foreign gene sequences matched with designed sequences, the insert sites were right, and the nucleotide mutation rate was less than one over ten thousands within different passages of rNTV-C; the target proteins could be expressed effectively, and the expression level was stable within different passages of rNTV-C; the genetic loss of gag and nef was less than 5% within different passages of rNTV-C. The above results provided important data for the vaccine production.
Animals ; DNA, Recombinant ; genetics ; Fibroblasts ; metabolism ; virology ; Gene Expression ; Genes, Viral ; genetics ; Genetic Engineering ; methods ; Genetic Vectors ; genetics ; HIV-1 ; genetics ; Sequence Analysis, DNA ; Vaccinia virus ; genetics