Nowadays,small peptides are always expressed in the form of fusion protein.The expression product contains many superfluous amino acids which can affect the biological functions of small peptides even expressed by GST fusion protein expression system.SUMO protease can cut SUMO fusion protein expressed by fusion expression system without any amino acid residues left on target protein thus become a hot topic in this field.Recombinant His-UlP1/pET3c/BL21(DE3)engineering strain was constructed by genetic engineering technology and the expression conditions were optimized in shake flaks.The process of high density fermentation was explored and different purification conditions were detected by chromatography.The results showed that SUMO protease could be expressed well after inducing the engineering strain by IPTG of 1.0mmol/L at 30℃ for 6 hours.The expression level of the strain in fermentation pots could reach 24.3% analyzed by SDS-PAGE.The purity of SUMO protease was more than 98% after further purification by cation exchange chromatography.The yield was 355mg SUMO protease per liter fermentation liquid.Western blot analysis demonstrated that there were immune reactions between IlP1 and 6?His antibodies,so it has established a good foundation for large-scale industralazation in the future.

OBJECTIVESTo study the effectiveness of an artificial liver support system.

METHODSThirty-two patients with medicamentous liver insufficiency were treated with an artificial liver support system in addition to the routine medicinal therapy. Thirty patients treated with routine medicinal therapy only served as controls.

RESULTSThe clinical symptoms (e.g. hepatic encephalopathy) and the laboratory indices (serum total bilirubin and prothrombin time) of the treatment group patients were obviously improved compared with those of the control group patients (P < 0.05). The cure rate and hospitalization days were 90.6% (26/32) and 47 days respectively in the treatment group, and 43.3% (13/30) and 72 days in the control group (P < 0.05).

CONCLUSIONUsing an artificial liver support system combined with routine medicinal therapy is more effective than using medication alone.

Adult ; Aged ; Antineoplastic Agents ; adverse effects ; Antitubercular Agents ; adverse effects ; Female ; Hepatic Insufficiency ; chemically induced ; therapy ; Humans ; Liver, Artificial ; Male ; Middle Aged

OBJECTIVETo investigate the relationship between the gene polymorphism of metabolizing enzymes and the genetic susceptibility to lung cancer as well as to study the synergistic effects between smoking and the genes.

METHODSA case-control study (case = 217, control = 200) was carried out to compare the frequent distribution of CYP1A1, 2E1, 2D6 and GSTM1 genotypes between the lung cancer group and the control group with a polymerase chain reaction-restriction fragment polymorphism (PCR-RFLP) method and to analyze the relationship between these genes and smoking.

RESULTSGSTM1-null genotype frequency was 58.5% in the lung cancer group and 47.5% in the control group with significant difference (P = 0.02). The frequent distribution of CYP1A1, 2E1, 2D6 genotypes was not significantly different in the two groups (P > 0.05). Synergistic effects were found between smoking and GSTM1 but not between smoking and CYP1A1, 2E1, 2D6.

CONCLUSIONSmoking and GSTM1-null genotype seemed to be the risk factors of lung cancer. Those who carrying GSTM1-null genotype and smoking cigarettes were prone to suffer from lung cancer to become the high-risk population of the disease.

Cytochrome P-450 CYP1A1 ; genetics ; Cytochrome P-450 CYP2D6 ; genetics ; Genetic Predisposition to Disease ; genetics ; Glutathione Transferase ; biosynthesis ; genetics ; Homozygote ; Humans ; Lung Neoplasms ; genetics ; Male ; Polymorphism, Genetic

Objective To observe the sequence of fat deposit and its relationship with insulin resistance in SD rats fed by high fat diet.Methods Normal 8-week-old male SD rats were randomly divided into normal chow (NC,n=40)and high fat diet(HF,n=40)groups.Triglyceride(TG)in serum,liver and muscle were measured;glucose infusion rate(GIR)and the mRNA level of genes related to lipid metabolism in liver and muscle were determined in different stages.GIR was detected by eugiyeemic-hyperinsulinemic clamp for evaluating the insulin sensitivity.Gene expression was determined by real-time PCR.Results(1)As compared with NC group,serum TG was not increased after high fat feeding for4 and 8 weeks,it began to increase after 12 weeks [0.52(0.15-1.00) mmol/L vs O.31(0.09-0.53)retool/L, P0.05)in skeletal muscle.After 8 weeks,the expression of ACC1 in liver in HF group was increased by 20.6%,CPT-1 was decreased by 27.1%(P

Adolescent ; Adult ; Aged ; Chronic Disease ; Female ; Hepatitis, Viral, Human ; complications ; therapy ; Humans ; Liver Failure ; etiology ; therapy ; Liver, Artificial ; Male ; Middle Aged

OBJECTIVESTo evaluate the relationship between the functional integrity of sperm membrane and seminal parameters related to CASA.

METHODSThirty-eight fertile and one hundrend and twenty four infertile males were tested the functional integrity of sperm membrane by the kit and parameters by CASA.

RESULTSThere was a significant difference in the functional integrity of sperm membrane between fertile and infertile group (P < 0.01). The items related to CASA between normal and abnormal group in the functional integrity of sperm membrane had a remarkable difference, except motion degree, seminal volume and pH.

CONCLUSIONSTo determine the functional integrity of sperm membrane can be used as a necessary supplementary method for CASA, and it has clinical significance in diagnosing, treating and researching male infertility.

Adult ; Humans ; Image Processing, Computer-Assisted ; Male ; Membranes ; physiology ; Semen ; physiology ; Spermatozoa ; physiology

This study was aimed to explore the role of human fetal bone marrow stromal cells (FBMSC) in combination with exogenous cytokines in supporting the in vitro expansion of cord blood mononuclear cells and the expression of CXCR4(+) and CD49d(+) in CD34(+) cells. Mononuclear cells (MNC) separated from cord blood (CB) were cultured in a serum-free support culture system with FBMSC or exogenous cytokines or both of them. On day 0, 6, 10 and 14, total cells were counted, CD34(+), CD34(+)CXCR4(+) and CD34(+)CD49d(+) cells were quantitated by FACS, and hematopoietic progenitor cells were assessed by semisolid culture assay. The results showed that after culturing for 14 days, CD34(+) cells, CD34(+)CXCR4(+) cells, CD34(+) CD49d(+) cells and colony forming unit (CFU) were significantly increased (P < 0.05). Compared with other groups, expansion multiple of CD34(+), CD34(+)CXCR4(+), CD34(+)CD49d(+) cells and CFU were higher than that in FBMSC and cytokine group (P < 0.05). It is concluded that the culture system used in this study can not only support the expansion of CB MNCs but also increase the number of hematopoietic stem and progenitor cells which has chemokine and adhesion capacity. This culture system may be a feasible way for in vitro culture of cord blood cells.
Antigens, CD34 ; blood ; Bone Marrow Cells ; cytology ; immunology ; Cell Proliferation ; drug effects ; Cells, Cultured ; Coculture Techniques ; Cytokines ; pharmacology ; Fetal Blood ; cytology ; Fetus ; Flow Cytometry ; Granulocyte-Macrophage Colony-Stimulating Factor ; pharmacology ; Humans ; Integrin alpha4 ; blood ; Interleukin-3 ; pharmacology ; Leukocytes, Mononuclear ; cytology ; immunology ; Receptors, CXCR4 ; blood ; Stromal Cells ; cytology ; immunology ; Time Factors

OBJECTIVETo study the mechanism involved in the control of glioma cell proliferation with transfection of connexin (Cx) 43 gene.

METHODSC6 rat glioma and TJ905 human glioblastoma cell lines without Cx43 gene expression were transfected with Cx43cDNA mediated by lipofectamine. Northern blot, in situ hybridization and immunohistochemical technology were used to detect the expression of Cx43mRNA and its protein with MTT assay and silver colloid stain for the detection of cell proliferation, TUNEL method for determination of cell apoptosis, scrape loading dye transfer (SLDT) for GJIC, Western blot and immunohistochemical technology for bFGF, PDGF, EGFR, IGF-I and IGFBP3 expression.

RESULTSCx 43 gene transfected glioma cells showed decreased proliferation, restored GJIC and decreased bFGF, PDGF, IGFBP3, except EGFR expression and cell apoptosis which showed no change.

CONCLUSIONThe mechanism of Cx 43 gene inhibiting gliomas cell proliferation is the restoration of GJIC and decreased autocrine growth factors.

Animals ; Apoptosis ; Cell Division ; physiology ; Connexin 43 ; genetics ; physiology ; DNA, Complementary ; genetics ; Glioma ; pathology ; Rats ; Transfection ; Tumor Cells, Cultured

We have constituted a mouse model for fetal blood transplantation (FBT) to cross over major histocompatibility complex (MHC) without causing serious GVHD. It seems that full matching at the MHC appears not necessary for FBT, while the nucleated cell dose is critical. Two fetal blood units were combined from different donors to increase the stem/progenitor cell dose so as to explore the possibility of MHC-mismatched allogeneic transplantation. 26 out of 40 mice in mixed FBT group survived in the observation period of 60 days after transplantation without obvious GVHD. Double chimerism was demonstrated by PCR and flow cytometric analysis; and skin transplantation test proved the induction of donor specific immune tolerance. Our data suggest that two MHC-mismatched allogeneic donor fetal blood units could simultaneously engraft and reconstitute immune and hematopoietic system in a mouse model. The result may be beneficial for the expansion of cord blood application and enables more patients to share the advantages of cord blood transplantation.
Animals ; DNA ; biosynthesis ; Female ; Fetal Blood ; immunology ; transplantation ; Graft vs Host Disease ; immunology ; mortality ; H-2 Antigens ; immunology ; Hematopoiesis ; immunology ; Hematopoietic Stem Cell Transplantation ; methods ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Survival Rate ; Transplantation Chimera ; genetics ; immunology ; Transplantation Tolerance ; immunology

OBJECTIVETo compare the bidirectional effect of rhubarb total anthraquinone (TA) and total tannins (TT) on rats' liver.

METHODSOne hundred rats were randomly divided into 10 groups, i.e., the blank group, the model group, the blank + high dose TA group, the blank +low dose TA group, the blank + high dose TT group, the blank + low dose TT group, the model + high dose TA group, the model + low dose TA group, the model +high dose TT group, and the model + low dose TT group, 10 in each group. The carbon tetrachloride (CCI4) was used to prepare the acute liver injury rat model. TA and TT of rhubarb (at 5.40 g crude drugs/kg and 14.69 g crude drugs/kg) were intragastrically administrated to rats in all groups except the blank group and the model group, once daily for 6 successive days.The general state of rats, biochemical indices such as alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), laminin (LN), hyaluronic acid (HA), transforming growth factor beta1 (TGF-beta1), as well pathological results of rat liver tissues. Finally the protection laws of TA and TT for rats' liver were analyzed using factor analysis.

RESULTSCompared with the blank control group, all biochemical indices increased in the blank group (P < 0.05, P < 0.01). HA also increased in the blank + high dose TA group; AST, ALT, and HA also increased in the blank +high dose TT group (P < 0.05). Compared with the model group, AST, ALT, ALP, HA, and TGF-beta1 significantly decreased in the model + low dose TA group, the model + high dose TA group, the model + low dose TT group (P < 0.05, P < 0.01). Serum AST, ALT, and ALP also decreased in the model + high dose TT group (P < 0.05, P < 0.01). Pathological results showed that mild swollen liver cells in the model + high dose TA group. Fatty degeneration and fragmental necrosis around the central veins occurred in the blank + high dose TA group. The pathological injury was inproved in the model +low dose TA group. Two common factors, liver fibrosis and liver cell injury, were extracted by using factor analysis. TA showed stronger improvement of the two common factors than TT.

CONCLUSIONSRhubarb TA and TT showed protective and harmful effects on rats' liver. At an equivalent dosage, TA had better liver protection than TT. High dose TT played a role in liver injury to some extent.

Animals ; Anthraquinones ; adverse effects ; pharmacology ; Carbon Tetrachloride ; toxicity ; Chemical and Drug Induced Liver Injury ; drug therapy ; pathology ; Dose-Response Relationship, Drug ; Female ; Liver ; drug effects ; Male ; Rats ; Rats, Sprague-Dawley ; Rheum ; chemistry ; Tannins ; adverse effects ; pharmacology