B-Lymphocytes ; Graft vs Host Disease ; Hematopoietic Stem Cell Transplantation ; Humans

This study was aimed to clone human intercellular adhesion molecule-1 (ICAM-1) gene, to transfect the constructed eukaryotic expression vector ICAM-1-GFP into CHO cells, as well as to detect ICAM-1-GFP expression in CHO cells binding with Molt-4 cells. ICAM-1 cDNA gene was amplified by RT-PCR and inserted in PMD(18)-T vector. Then ICAM-1 cDNA from pMD18-ICAM-1 vector was subcloned into eukaryotic expression vector pEGFP-C1 to construct recombinant ICAM-1-pEGFP-C1 vector. Restriction analysis and DNA sequencing were used to confirm the recombinant vector. After stable transfection of CHO-K1 cells with the recombinant vector, the expression and subcellular localization of ICAM-1-GFP were detected by RT-PCR, flow cytometry and fluorescence microscopy. The function of ICAM-1-GFP fusion protein was assessed by the binding of ICAM-1-GFP/CHO cells to Molt-4 cells. The results showed that 1622 bp full-length ICAM-1 cDNA obtained and was successfully ligated with pMD(18)-T-vector, subcloned to construct recombinant ICAM-1-pEGFP-C1 vector. Restriction analysis and DNA sequencing indicated that recombinant ICAM-1-GFP was successfully constructed and ICAM-1-GFP was expressed stably in CHO cells. ICAM-1-GFP expression was only observed in the cytoplasm of ICAM-1-GFP/CHO cells by fluorescence microscopy. The ICAM-1-GFP/CHO cells were bound to PMA-treated Molt-4 cells. The expression of MEM-148 was very weak in PMA-treated Molt-4 cells. It is concluded that the ICAM-1-GFP eukaryotic expression vector has been constructed successfully and expresses stably in CHO cells. PMA can increase the binding of Molt-4 cells to ICAM-1-GFP/CHO cells by inducing specialized form of ICAM-1 clustering.
Animals ; CHO Cells ; Cricetinae ; Cricetulus ; DNA, Complementary ; genetics ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; Humans ; Intercellular Adhesion Molecule-1 ; genetics ; Recombinant Proteins ; genetics ; Transfection

After allogeneic hematopoietic stem cell transplantation (allo-HSCT), the donor cells present a profound immunization therapy efficiency. Among these effector cells, allo-reactivity natural killer (NK) cell activation are concerned with the graft-versus-host disease (GVHD) and graft-versus-leukemia (GVL) effect. As known, GVHD is primarily a T-cell-mediated event but not initiated by NK cells. NK cells may significantly enhance GVL immune response by using an integration of activating and inhibitory receptors. Allo-reactivity NK cell infusion after allo-HSCT already transits from experiments to clinic. In this review the background on NK cells, and their clinical roles in Allo-HSCT were summarized.
Graft vs Host Disease ; immunology ; Graft vs Leukemia Effect ; immunology ; Hematopoietic Stem Cell Transplantation ; Humans ; Killer Cells, Natural ; immunology ; Transplantation Immunology

To explore feasibility of real-time quantitative PCR in assessing efficiency of gene transfection, clonal PCR was employed to analyze efficiency of retroviral-mediated neo gene transfection in primary myoblast, simultaneous real-time PCR were performed for estimation of transfection efficiency; for measuring integrated gene copy number per cell, linear amplification mediated-PCR (LAM-PCR) and retroviral 5'LTR integration analysis also were used. The results showed that: (1) the data from clonal PCR are similar as that from real-time PCR in low efficiency of transfection (< 36%); but in high efficiency of transfection, it is significantly differentiation between clonal PCR and real-time PCR. (2) One copy of transduced gene per cell was observed in retroviral-mediated gene transfection in primary myoblast. It is concluded that real-time PCR can be used to estimate gene transfer vector in vivo, but it is not available for assessing gene transfection in vitro, because high efficiency of transfection could be obtained in most of gene transfection in vitro.
Animals ; Genetic Vectors ; genetics ; Polymerase Chain Reaction ; methods ; Retroviridae ; genetics ; Sequence Analysis, DNA ; Transfection

LFA-1 and ICAM-1 mediate a bi-directional signaling across the cell membrane which is essential for biological functions of lymphocyte, including exudation, activation, adhesion, immunosurveillance as well as immuno-logical synapse formation. The signal transducing is a dynamic process and dependent on the binding capacity between LFA-1 and ICAM-1. The affinity and the avidity of LFA-1 are two major regulation forms in this process. Phosphorylation of LFA-1 and cytoskeleton protein talin 1 play a critical role in signal transducing. In biology of lymphocyte, LFA-1 and ICAM-1 interaction forms the co-stimulatory signal to promote activation, proliferation and division. In this article the regulation of binding capacity between LFA-1 and ICAM-1, the regulation of LFA-1 subunit phosphorylation, the role of talin1 in signaling transduction of LFA-1 and ICAM-1, the synergic stimulatory signaling of LPA-1 and ICAM-1 were reviewed.
Humans ; Intercellular Adhesion Molecule-1 ; metabolism ; physiology ; Ligands ; Lymphocyte Function-Associated Antigen-1 ; metabolism ; physiology ; Lymphocytes ; cytology ; immunology ; metabolism ; Phosphorylation ; Signal Transduction ; physiology ; Talin ; metabolism

As well as playing vital roles in main cellular processes, such as abnormal proliferation, differentiation, survival, apoptosis, and a lot of tyrosine kinases (TK) are involved in oncogenesis. TK or components of their signal pathways have been found abnormal in many hematological malignancies. Therefore, tyrosine kinase inhibitors (TKI) have been provided a great deal of enthusiasm for development of therapy in myeloproliferative neoplasms (MPN). Representativity, the treatment of chronic myelogenous leukemia (CML) was revolutionary for the design of imatinib mesylate (IM), which is a BCR/ABL TKI. Subsequently, because of need for the resistance or intolerance, novel agents are being explored and imatinib has now been extended to eosinophilia-associated myeloid neoplasms with PDGFRA, PDGFRB or FGFR1 gene mutations. Recently, JAK2 inhibitor drugs are currently being tested in clinical trials. Here, the current review mainly focuses on the role of TK in classic MPN including CML, polycythemia vera (PV), primary myelofibrosis (PMF), essential thrombocythemia (ET), and advances of targeting these abnormalities with small molecule inhibitors.
Drug Delivery Systems ; Humans ; Myeloproliferative Disorders ; drug therapy ; Protein-Tyrosine Kinases ; antagonists & inhibitors ; metabolism

OBJECTIVETo investigate the cost-effectiveness of fish oil in patients undergoing major surgery and those with systemic inflammatory response syndrome(SIRS).

METHODSA retrospective study was conducted in patients undergoing major surgery and those with SIRS on admission in the Zhongshan Hospital from January 2008 to December 2011. Fish oil group was enrolled and matched to control group by 1:2 for gender, age, diagnosis, and surgical procedure. There were 220 pairs of patients who were not admitted to ICU, 102 pairs of patients admitted to ICU, and 66 pairs of patients with SIRS. The clinical outcomes and costs were measured and cost-effectiveness analyses were conducted.

RESULTSThe clinical outcomes and costs showed no significant difference between the fish oil group and the control group in those patients who were not admitted to ICU(P>0.05). Fish oil fat emulsion supplementation significantly reduced the length of total hospital stay, postoperative hospital stay, ICU stay, re-operation rate, infection rates, perioperative mortality in patients admitted to ICU and those with SIRS(P<0.05). The cost-effectiveness ratio of non-reoperation rate, non-infection rate, and survival rate were lower in those patients receiving fish oil fat emulsion as compared with those without fish oil administration. Fish oil fat emulsion supplementation could reduce cost-effectiveness ratios of non-reoperation rate, non-infection rate and survival rate by 105 RMB, 160 RMB, and 89 RMB respectively in major surgical patients who admitted to ICU, and by 670 RMB, 280 RMB, and 220 RMB respectively in SIRS patients.

CONCLUSIONSAddition of fish oil fat emulsion to clinical nutrition may have positive effects on critically ill patients. It seems that the effects of fish oil fat are strongly related to the severity of patient's underlying disease. Fish oil fat emulsion supplementation shows acceptable cost-effectiveness ratio and pharmacoeconomic value.

Aged ; Cost-Benefit Analysis ; Fat Emulsions, Intravenous ; economics ; therapeutic use ; Female ; Fish Oils ; economics ; therapeutic use ; Humans ; Male ; Middle Aged ; Parenteral Nutrition ; economics ; methods ; Postoperative Care ; Retrospective Studies ; Surgical Procedures, Operative ; Systemic Inflammatory Response Syndrome ; therapy

Air Pollutants, Occupational ; analysis ; Coke ; Fatty Liver ; epidemiology ; Humans ; Logistic Models ; Male ; Occupational Exposure ; adverse effects ; Risk Factors

Objective:To analyze the relationship between hepatitis B virus(HBV)gene mutation at 1896 in precore region with genotype and replication of HBV and the liver function of patients.Methods:HBV precore 1896 site mutation,the genotype of HBV and serum content of HBV DNA were determined by PCR in 60 patients positive of HBV DNA.Chemiluminescence miacropaticle immunoassay(CMIA)was used for detection of serum HBeAg and HBeAb.Liver function parameters were ob- tained by routine biochemistry method.Results:The alanine aminotransferase(ALT)level in HBV with 1896 site mutation was significantly higher than that in the wildtype virus.Site mutation at 1896 had no correlation with HBeAg,HBV genotype and HBV DNA content.HBV DNA content in patient with genotype C was significantly higher than that with genotype B(P

The changing of atherosclerosis (AS) plaque from stable to unstable is closely related to the incidence of cardio-cerebrovascular events. So, to stabilize AS plaque become the hot point of the modern study on prevention and treatment of cardio-cerebrovascular diseases. The formation of AS plaque is a very long and complicated process with the participation of many factors. However, inflammation reaction plays an important role in the formation and rupture of unstable plaque. Traditional Chinese medicine (TCM) shows latent therapeutic advantages in stabilizing AS plaque with its characteristics of multi-way, multi-link, and multi-target point and all-sided treatment as well as its rather milder adverse reactions. The paper gives a comparatively systematic retrospection of study on unstable plaque treated by integrated Chinese and Western medicine conducted in recent years. Combined with the related topics conducted by the authors, they raise the theory of "unstable plaque is induced by toxinduced by toxin and stagnation", and introduce the study on the intervention of unstable plaque by removing toxic substances and activating blood flow. They believe that the explorative study can provide objective experimental and clinical basis for TCM therapeutic method in stabilizing AS plaque.
Angina, Unstable ; drug therapy ; Coronary Artery Disease ; drug therapy ; Diagnosis, Differential ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Medicine, Chinese Traditional ; Phytotherapy