As an effective method of studying soluble protein-protein interactions, yeast display system is now widely used for affinity maturation of single-chain antibodies.Due to the strong homology recombination machinery of yeast and the high-throughput nature of FACS detection, a rapid scan for interaction between antigen-antibody pairs could be easily achieved.Based on this system, a novel and reliable method for determining conformational epitopes was developed.Different fragments of macrophage migration inhibitory factor(MIF) and several point mutations of MIF were displayed on yeast cell surface using homologous recombination technology.Three MIF monoclonal antibodies, 10C3, 2A12 and 4E10, were screened for their binding affinity to each displayed peptide.Utilizing this technology, the key amino acids of MIF that bind to the MIF monoclonal antibodies were easily identified.

OBJECTIVE: To investigate the protective effects of rosiglitazone maleate on renal ischemia-reperfusion injury in mice and its mechanism.METHODS: 30 mice were randomly divided into sham operation group(normal saline),model group(normal saline) and rosiglitazone maleate group(rosiglitazone maleate 5 mg?kg-1?d-1).Three groups received relevant drugs for 10 days.Renal ischemia-reperfusion injury model was induced in model group and rosiglitazone maleate group by clipping the renal pedicle 40 minutes after the last administration.The renal index,the content of blood urea nitrogen(BUN) and creatinine(Cr),the content of malondialdehyde(MDA),the activities of superoxide dismutase(SOD) and glutathione peroxidase(GSH-Px) in renal tissue were detected.Pathological change of renal tissue was observed by HE stain.RESULTS: Compared with model group,the renal index of the mice in rosiglitazone maleate group reduced(P

Objective To explore methods for establishinga cellu lar injury model of blood stasis syndrome (BSS). Methods Two experiments were c arried out. In the first experiment,the injured vascular endothelial cell model of BSS was established by the culture of vascular endothelial cell, and in the seco nd experiment, the vascular endothelial cell injured by the serum of BSS rabbit was cultured. Results In the first experiment, pathological features and end ocrine dysfunction in the primary cultured endothelial cell were similar to the B SS rabbit model, but the above changes did not remain in the subculture. The above changes were also found in the second experiment and were easy to be repea ted. Conclusion The injured vascular endothelial cell model of BSS establis hed b y the above methods reflects or partially reflects the structure and function of BSS rabbit model and the patients with BSS, and can be used to study the pathol ogical mechanism of BSS and therapeutic effect of blood_circulation activating a nd blood_stasis removing herbs.

Objective To investigate the vasodilatation effects of total flavonoid from polygonum aviculare on rat thoracic aorta and its underlying mechanisms. Methods Total flavonoid from polygonum aviculare was gotten by extracted with 65% alcohol, gathered with polyamide, and eluted with 75% alcohol.The content of flavone was determined with rutoside as standard preparation. Normal rats thoracic aorta in vitro used as sample, BL-420E biological functional experiment system was utilized to record dilatation effect of total flavonoid on PE affecting pre-contracting blood vessel and the relation between dilatation effect of total flavonoid on blood vessel and calcium influx. Results Total flavonoids from polygonum aviculare can diastole contractions of thoracic aorta caused by PE. In calcium-free perfusion, gradually adding CaCl2 induced calcium influx. Clinical data showed dose-effect relation between drug and blood vessel contraction decreased in the total flavonoids from polygonum aviculare incubation rats than normal rats. Besides, total flavonoids from polygonum aviculare can obviously inhibit contraction of blood vascular circle induced by calcium releasing.Conclusion FP exerted a dose-dependent vasodilatation effect on rat isolated aorta rings by inhibiting Ca2+influx via L-type voltage-gated Ca2+ channels and Ca2+ release from sarcoplasmic reticulum, consequently decrease Ca2+ in vascular smooth muscle cells.

Objective To explore the relationships of depression status of medical college students with their automatic thoughts and personality. Methods A total of 1 440 medical college students were tested by self-rating depression scale (SDS), automatic thoughts questionnaire (ATQ) and Eysenck personality questionnaire (EPQ). Results ①27.5% medical college students had different degree depressive mood. ② Compared with the non-depression group students, the depression group had higher scores in ATQ, EPQ-N(neurosism) and EPQ-P(psychoticism), lower scores in EPQ-E(extroversion), EPQ-L(lie) (P

Objective To study the influence of the different tidal volume on hemodynamics,pulmonary ventilation and pulmonary mechanics during mechanical ventilation in patients with acute lung injury.Methods To adopt contrast method,making use of the VIIP +0.196 kPa of P-V curve to decide PEEP,then according to the VUIP of P-V curve,obtaining the three different data of humidity 100%Vt,85% Vt and 70 Vt.At the same time,monitoring the change of static compliance,the changing of hemodynamics and P-V curve.Results 85%Vt group was better than 100%Vt group and 70%Vt group in the heart rates,CVP,PaO_2,the peak airway pressure(PIP) and mean pressure(Pm),Cst,PaO_2/FiO_2.Conclusions When insuring the PEEP data in low inflection point pressure (Pinf) of breathing system P-V curve,and 85% humidity in upper inflection point pressure,which is in accordance with individual protecting breathing strategy.The curative effect is the best for improving static compliance and decreasing the harm of lung injury.The change of pressure-volume curve should be paid attention to in nursing.

OBJECTIVE To learn the disinfection and sterilization for dental instruments and to provide scientific basis for related regulations according to the scheme made by Beijing Center for Disease Prevention and Control.METHODS To investigate some dental medical units by adopting investigation form,inquirement,checking about the record for disinfection equipment including such items as air,the surface of used material,hands,etc.RESULTS Altogether 39 dental medical units including 9 hospitals,21 joint clinics and 9 private medical units were investigated with 96.11%,88.79%,and 70.97% qualified rates,respectively.CONCLUSIONS In private dental units the qualified rate for disfection is poor,disinfection instruments are short,and the professionals knowledge related to disfection is little.As a result,strengthening monitoring and guidance to prevent hospital infection should be carried out at once.

Objective To observe the protective effect of Xuefu Zhuyu Decoction (XZD) on vascular endothelial cells (VEC) injured by the serum of blood_stasis_syndrome ( BSS) rabbits. Methods The morphological features of VEC cultured with norma l rabbit serum (Group A), BSS rabbit serum(Group B), the serum of normal rabbit medicated with XZD(Group C) and the serum of BSS rabbit medicated with XZD(Group D) were observed respectively under light microscope and electron microscope. Results Under light microscope, VEC was shrinked, intercellular space widene d and cytoplasm contained dark granules in Group B. The change of intercellular space was not obvious in Group D. Under electron microscope, pinocytotic vesicle s increased, rough endoplasmic reticulum (RER) decreased and dilated, mitochondr ia became smaller and indistinct and only a small amount of microvilli appeared on cell membrane with top expanded and merged in VEC of Group B as compared with Group A and Group C. Less RER but no dilatation and fewer microvilli without to p merged were found in VEC of Group B. Conclusion The serum of BSS rabbits c an injury normal VEC cultured in vitro and XZD exerts a certain protective effe ct on VEC injured by the serum of BSS rabbits.

BACKGROUND:Chinese herb, bushen yizhi formula protects at certain extent learning and memory in rat model of Alzheimer disease. The drug serum in this formula can alleviate neurotoxic reaction of nerve tumor cell NG 108-15 to beta-amyloid protein. In order to understand further the mechanism and compatibility of the formula, it is necessary to carry on the study on the broken formulas.OBJECTIVE:To study the effect of drug serum in subgroups of broken bushen yizhi formulas on growth and differentiation of cell model of Alzheimer disease and probe into the compatibility rule of bushen yizhi formula in view of serum pharmacology.DESIGN: Randomized controlled experiment.SETTING: DME Center of Clinical Pharmacological Institute Affiliated to Guangzhou University of Traditional Chinese Medicine.PARTICIPANTS:The experiment was performed in DME Center of Clinical Pharmacological Institute Affiliated to Guangzhou University of Traditional Chinese Medicine from January to August 2003, in which, 40 healthy male SD rats of 3 months old were employed and NG108-15 cell line was frozen-preserved.into the control, original formula group (No. 1 group) [shechuanzi (Cnidium monnieri (L.) Cuss), gouqizi (Lycium barbarum L.), renshen (Panax ginseng C.A.Mey.), heshouwu (Polygonum multiflorum Thunb.), danpi (Paeonia Suffruticisa Andr.) and bingpian (Borneolum)], kidney replenishment group (No.2 group) [shechuanzi (Cnidium monnieri (L.) Cuss), gouqizi (Lycium barbarum L.), etc.], group for benefiting qi and nourishing blood (No.3 group)[renshen (Panaz ginseng C.A.Mey), zhishouwu (Polygonum multiflorum Thunb.), etc.] and group with bingpian (Borneolum) removed (No.4 group)[Panax ginseng C.A.Mey.], heshouwu (Polygonum multiflorum Thunb.) and danpi (Paeonia Suffruticisa Andr.)], 8 rats in each group. The concentrated Chinese herbal solutions of every group were applied at 10 μL/g (equal to 6 g/kg of raw herbs) for gastric infusion successively,continuously for 1 month. In the control, the physical saline solution of equal dosage was used for infusion. Two hours after the last gastric infusion in rats of each group,the blood was collected from heart after anesthesia and the serum was sepaNG108-15 cell cultured in vitro was divided into 6 groups. In the control and model group, normal rat serum was contained in proliferated culture solution. In the rest 4 groups, the drug serum of No. 1 group and 3 sub-groups was contained.Simultaneously, beta-amyloid protein 25-35 in each hole was prepared to the terminal concentration 5 μmol/L (except in the control) and the culture went on for 48 hours.MAIN OUTCOME MEASURES:MTT method was used to determine proliferated number and survival rate of cells. Simultaneously, the ratio of neurite cells to total cell count and average length of neurit were determined.icantly than the control (0.520±0.022, 0.665±0.037, P ＜ 0.01), and that in every drug serum group was higher than model group, of which, the result vival rate of differentiated cells: That in model group was lower significantly than the control (58.4%, 100%) and that in every drug serum group was higher than model group, of which, the result in No.4 group was the most tal cell count: That in model group was lower significantly than the control [(42.95±11.42)%, (58.75±12.84)%, P ＜ 0.01] and that in every drug serum group was higher than model group, of which, the result in No.4 group was rite: That in model group was shorter significantly than the control [(356.0 ±109.0), (493.8±133.0) μm, P ＜ 0.01] and that in every drug serum group was longer than model group, of which, the result in No.4 group was the most significant [(486.8±79.2) μm, P ＜ 0.01].CONCLUSION: The drug serum in all of bushen yizhi formula and every subgroup inhibits at certain extent the injury of beta-amyloid protein 25-35 to NG108-15 cell, but the results of each group are various. The protection of drug serum to the cell in every group is in the sequence from strong to weak as group with bingpian removed ＞ original formula group ＞ kidney replenishment group ＞ group for benefiting qi and nourishing blood. It is to expect a further study on the efficacy of group with bingpian removed.

ObjectiveTo investigate the effects of glioma microenvironment and COX-2 inhibitor celecoxib on the phenotypes and function of dendritic cell (DC).MethodsThe expression of cyclooxygenase-2 (COX-2) and prostaglandin E2 (PGE2) production were detected in glioma C6 cells treated with different concentration of celecoxib.Monocytes were isolated from human peripheral blood and cultured with 200 ng/ml rhGM-CSF and 50 ng/ml rhIL-4,either C6 tumor cells supernatant (TSN) or TSN from C6 cells treated with celecoxib to generate DCs.Cell morphology was observed.Cell phenotype including CD1a,CD80,CD83 and D86 were analyzed on a FACScan.Production of IL-12 in DC supernatant and the potential to stmiulate allogeneic T cell proliferation were detected.ResultsThe expression of COX-2 and PGE2 production in C6 cells decreased after treated with celecoxib in a concentration dependant manner.Typical DCs were induced in all groups and the expression of CD1a ((75.56±2.40)％,(75.09±3.67)％,(76.03 ±3.43)％),CD83((72.04±3.45)％,(71.44±3.78)％,( 73.63 ± 3.31 ) ％ ) had no difference (P ＞ 0.05 ).Expression of CD80 ( ( 58.41 ± 3.85 ) ％ ),CD86 ( ( 58.22 ±3.25)％ ) in DC with TSN obviously decreased compared with normal group( (70.36 ± 2.91 )％,(69.31 ±4.29 ) ％,P ＜ 0.01 ) as well as the IL-12 production ( ( 137.88 ± 5.33 ) pg/ml,( 186.04 ± 4.76 ) pg/ml) and the potential to stmiulate allogeneic T cell proliferation ( P ＜ 0.01 ).Celecoxib increased the expression of CD80,CD86 (66.83 ± 2.51,63.51 ± 5.47,P＜ 0.01 ) in DC and the same as IL-12 production( ( 170.31 ± 3.46) pg/ml,P ＜ 0.01 ) and the potential to stmiulate allogeneic T cell proliferation ( P ＜ 0.01 ),which were lower than the normal level.ConclusionGlioma microenvironment may induce the celecoxib can inhibit the expression of COX-2 and PGE2 in gliomas cells and improve the phenotypes and function defect of DCs.