ABSTRACT
To investigate the effects of human anti-BAFF scFv-Fc against the hsBAFF, ICR mice were randomly divided into six groups: control, hsBAFF (1 mg x kg(-1)), hsBAFF (1 mg x kg(-1)) + Ab (1 mg x kg(-1)), hsBAFF (1 mg x kg(-1)) + Ab (2 mg x kg(-1)), hsBAFF (1 mg x kg(-1)) + human IgG (1 mg x kg(-1)) and hsBAFF (1 mg x kg(-1)) + human IgG (2 mg x kg(-1)) groups. The effects of scFv-Fc administration on the proliferation of B lymphocytes were evaluated using an MTT assay. The titres of antibody in the serum and B lymphocytes differentiation were assessed by ELISA and flow cytometry, respectively. The results showed that administration of scFv-Fc to mice injected with hsBAFF significantly prevented human BAFF-induced increases in splenic B cell numbers and serum immunoglobulin levels. Furthermore, this fully human antibody would avoid inducing the human anti-mouse antibody (HAMA) response when used in humans. These findings suggest that the compact antibody may be useful in therapeutic or diagnostic application of the BAFF-associated autoimmune diseases in human.
Subject(s)
Animals , Female , Humans , Mice , B-Cell Activating Factor , Allergy and Immunology , Metabolism , B-Lymphocytes , Cell Biology , Body Weight , Cell Differentiation , Cell Proliferation , Cells, Cultured , Immunoglobulin Fc Fragments , Allergy and Immunology , Metabolism , Immunoglobulin G , Blood , Allergy and Immunology , Immunoglobulin M , Blood , Mice, Inbred ICR , Random Allocation , Recombinant Fusion Proteins , Allergy and Immunology , Metabolism , Single-Chain Antibodies , Allergy and Immunology , Metabolism , Spleen , Cell BiologyABSTRACT
<p><b>OBJECTIVE</b>To investigate the relationship between apolipoprotein e4 allele and emergence agitation (EA) in patients undergoing general anesthesia.</p><p><b>METHODS</b>A nested cohort study was conducted in elderly patients (over 60 years old) scheduled for major abdominal surgery requiring general anesthesia. A structured interview was conducted in PACU to determine EA, defined using the Sedation-Agitation Scale (SAS). Blood samples were obtained for measurement of the apolipoprotein genotypes.</p><p><b>RESULTS</b>Of the 196 patients studied, 22.4% developed EA. Thirty-eight patients (19.4%) had the apolipoprotein e4 allele. The presence of the e4 allele and low level of education were both associated with an increased risk of EA (36.9% vs15.8%, P=0.005; 30% vs 14.3%, P=0.01). After adjustment for covariates, the patients with the copy of e4 allele were shown to have a greater likeliness of an increased risk of EA (odds ratio: 4.32; 95% CI: 1.75-10.05) than those without the e4 allele.</p><p><b>CONCLUSION</b>Apolipoprotein e4 carrier status is associated with an increased risk for EA.</p>
Subject(s)
Aged , Female , Humans , Male , Middle Aged , Abdomen , General Surgery , Alleles , Anesthesia Recovery Period , Anesthesia, General , Methods , Apolipoprotein E4 , Genetics , Genotype , Psychomotor Agitation , Genetics , Risk FactorsABSTRACT
<p><b>OBJECTIVE</b>To observe the effect of huperzine A on the cognitive function of rats recovering from general anesthesia and discuss its possible mechanism.</p><p><b>METHODS</b>Sixty rats (20 to 23 weeks old) were subjected to spatial reference memory version of navigation task, in which the rats were expected to locate the escape platform in water. Two sessions of training were given daily for 5 days, and on the 5th day, the escape latencies of the rats were recorded. The rats were then divided randomly into 5 groups (n=12), and in 4 of the groups, the rats received an intraperitoneal injection of diprivan (Dip) at 2 mg/kg and after recovery of righting reflex, huperzine A was given at 0.05 mg/kg (group L), 0.1 mg/kg (group M), 0.2 mg/kg (group H), and in group C, no subsequent huperzine A was given; in group E, the rats received normal saline injection only. One hour after righting reflex recovery, the escape latencies of all the rats were recorded again, and the level of AChE expression in the forebrain cortex was measured quantitatively.</p><p><b>RESULTS</b>The escape latencies after righting reflex recovery was significantly longer than that on day 5 (P<0.05), and the rats in group H had the shortest escape latency among the groups (P<0.05). The average gray scale of AChE in the forebrain of rats in group H was significantly lower than that of the other groups (P<0.05).</p><p><b>CONCLUSION</b>Huperzine A can inhibit cholinesterase in the brain to improve the cognitive function of rats recovering from general anesthesia.</p>
Subject(s)
Animals , Rats , Alkaloids , Anesthesia, General , Cholinesterase Inhibitors , Pharmacology , Cognition , Maze Learning , Propofol , Rats, Sprague-Dawley , Sesquiterpenes , PharmacologyABSTRACT
<p><b>AIM</b>To investigate effects of hsBAFF synthesized in Escherichia coli on spleen B lymphocyte immune response and its intracellular free Ca2+ ([Ca2]i]) signaling in mice.</p><p><b>METHODS</b>Twenty ICR mice, half males-half females, were chosen and randomly divided into a normal control group (n=10) and a hsBAFF treatment group (n-10). The mice in hsBAFF treatment group were given abdominal cavity injection of hsBAFF solution which was diluted with phosphate buffered saline (PBS) at dosage of 0.1 mg/kg body weight once each day for over eight days. The mice in control group were received abdominal injection of PBS at the same dose and frequency. Spleen B lymphocyte proliferation and its immune response to LPS stimulation in mice were evaluated using an MTT assay, and change of spleen B lymphocyte [Ca2+]i was assayed under a laser scanning confocal microscope.</p><p><b>RESULTS</b>B lymphocyte proliferation and its immune response to LPS stimulation were significantly higher in hsBAFF-treated mice than in control mice (P < 0.05). The B lymphocyte [Ca2+]i fluorescence intensity in hsBAFF-treated mice maintained at a relatively high level fluctuation, and its average intensity was significantly higher to that of control mice (P < 0.01), but change rate of the intensity was lower compared to that of control group.</p><p><b>CONCLUSION</b>hsBAFF synthesized in Escherichia coli can enhance immune function in the body by increasing B lymphocyte proliferation and its immune response. hsBAFF-activated B lymphocyte function may be associated with increasing B lymphocytes [Ca2+]i.</p>
Subject(s)
Animals , Female , Male , Mice , B-Cell Activating Factor , Allergy and Immunology , Pharmacology , B-Lymphocytes , Cell Biology , Allergy and Immunology , Calcium , Metabolism , Calcium Signaling , Cell Proliferation , Mice, Inbred ICR , Spleen , Cell Biology , Allergy and ImmunologyABSTRACT
BCMA is one of the transmembrane receptors belonging to BAFF and APRIL. In order to identify the feasibility of sBCMA as decoy receptor and obtain active sBCMA for its structural and functional research, full length of hBCMA was amplified with total RNA from Raji cell line by RT-PCR, and the cDNA encoding the extracelluar soluble domain of hBCMA was inserted into pET43.1a(+) vector. The recombinant vector pET43.1a(+)-sBCMA was transformed into E. coli Origami B(DE3) pLyS which is helpful for disulfide bond construction of expression proteins. After IPTG induction, the recombinant protein was expressed as soluble fusion protein, sBCMA-NusA-His6, and identified by western blotting. Then the target protein was purified by Ni(+)-chelating Sepharose Fast Flow. The binding activity between recombinant sBCMA and BAFF was detected by ELISA. Also, Recombinant sBCMA inhibited proliferation of mouse B cell stimulating by rhsBAFF. It was proved that recombinant sBCMA has good bioactivity and the method to express those proteins rich in disulfide bond is feasible and effectual.
Subject(s)
Humans , B-Cell Activating Factor , Chemistry , B-Cell Maturation Antigen , Genetics , Cloning, Molecular , DNA, Complementary , Genetics , Disulfides , Chemistry , Escherichia coli , Genetics , Metabolism , Recombinant Fusion Proteins , Genetics , Solubility , Tumor Necrosis Factor Ligand Superfamily Member 13 , ChemistryABSTRACT
<p><b>OBJECTIVE</b>To observe the effect of huperzine A on cerebral cholinergic system in elderly patients during recovery from general anesthesia.</p><p><b>METHODS</b>Thirty elderly patients undergoing elective surgery under general anesthesia were randomized in a double-blind manner into group I (n=15) to receive huperzine A (0.3 mg/2 ml) and group II (n=15) with normal saline (2 ml) given intravenously. Huperzine A or normal saline was administered 30 min before completion of the operation, and acetylcholine (Ach) concentration in the cerebral spinal fluid (CSF) of the patients was determined using high-performance liquid chromatography with electrochemical detector (HPLC-ECD) and the activity of cholinesterase inhibitors (ChE) evaluated with automatic biochemistry analyzer before general anesthesia induction (T1) and 5 h after operation completion (T2).</p><p><b>RESULTS</b>In both the groups, Ach concentration in the CSF were lower at T2 than that at T1 (P<0.01), and at T2, CSF Ach concentration was significantly higher in group I than in group II (P<0.01); in group I, the activity of CSF ChE at T2 was lower than that at T1 (P<0.01), and also lower than at T2 in group II (P<0.01).</p><p><b>CONCLUSION</b>Huperzine A can inhibit cholinesterase to increase Ach, which has a positive effect on cerebral cholinergic system in elderly patients during recovery from general anesthesia.</p>
Subject(s)
Aged , Female , Humans , Male , Acetylcholine , Cerebrospinal Fluid , Alkaloids , Anesthesia Recovery Period , Anesthesia, General , Brain , Metabolism , Cholinesterase Inhibitors , Therapeutic Uses , Cholinesterases , Cerebrospinal Fluid , Chromatography, High Pressure Liquid , Double-Blind Method , Electrochemistry , Intraoperative Period , Sesquiterpenes , Therapeutic UsesABSTRACT
<p><b>AIM</b>To investigate the effect of epigallocatechingallate (EGCG) on acute lung injury induced by oleic acid in mice and the possible mechanism.</p><p><b>METHODS</b>Acute lung injury was induced by oleic acid in mice. Light microscopy and electron microscopy were used to examine histological changes and lung index as well as wet to dry weight ratio was calculated. Serum TNF-a level was measured by enzyme linked immunosorbent assay (ELISA) and the phosphorylation of p38 MAPK was determined by Western blotting.</p><p><b>RESULTS</b>Pretreatment of EGCG significantly alleviated oleic acid induced lung injury accompanied by reduction of lung index and wet to dry weight ratio, decreased of TNF-a level in serum and inhibition of phosphorylation of p38 MAPK.</p><p><b>CONCLUSION</b>EGCG showed beneficial effect on acute lung injury induced by oleic acid in mice. The ultimate reduction of TNF-alpha in serum caused by inhibition of phosphorylated p38 MAPK is involved in the mechanism of action of EGCG.</p>
Subject(s)
Animals , Male , Mice , Catechin , Pharmacology , Lung , Pathology , Oleic Acid , Phosphorylation , Protective Agents , Pharmacology , Respiratory Distress Syndrome , Metabolism , Pathology , Tumor Necrosis Factor-alpha , Metabolism , p38 Mitogen-Activated Protein Kinases , MetabolismABSTRACT
The prokaryotic expression plasmid pET-30a(+)/sBLyS was constructed and transformed into E. coli BL21 (lambda DE3). The recombinant protein was found to be highly expressed by the plight of soluble part and inclusion body. For the sake of enhancing the proportion of the soluble part, inducement at 16 degrees C for 12 h was ascertained. The expressing product was then purified by Ni2+ affinity chromatography gel. PI of the recombinant human sBLyS(rhsBLyS) is about 7.1-7.3 and it assembles into a homotrimer. The effect of rhsBLyS on B lymphocytes by MTT method told us the B lymphocytes' proliferating capacity dose depended on concentration and also stimulating time of the rhsBLys. With rhsBLyS(2 micrograms/mL) stimulating 3 days, B lymphocytes can proliferate the most.