Objective To induce and culture purity dendritic cells (DCs) of different maturation phase from human peripheral blood monocytes in vitro. Methods The monocytes were acquired from human peripheral blood mononuclear cells（PBMC）by adhesion method.The first step was to work on a 7-day culture of monocytes in medium supplement with GM-CSF and IL-4, and the cells were actually immature. In the second step,the cells were cultured in the medium supplement with GM-CSF and TNF-α,and cultured until the 14th day. Mature DCs were obtained. Cells harvested were identified for their morphology by microscope, properties of DCs detected by FCM, function with MTT method. Results Immature dendritic cells expressed CD1a molecules and costimulating molecules in middle level, HLA-DR in high level. In maturation phase DCs expressed costimulating molecules, HLA-DR molecules, CD83, and CD25 highly. These DCs could stimulate proliferation of allogenic T lymphocytes and increase killing ability of CTL activated by DCs. Conclusion The method to obtain DCs of different maturation phases from human peripheral blood monocytes is successfully set up. A number of DCs with high purity are obtained.

Objective To investigate the role of peroxisome proliferator-activated receptor-γ(PPAR-γ) activation in rat mesangial cell extracellular matrix（ECM）accumulation induced by advanced glycation end products (AGEs). Methods Rat mesangial cells were treated with AGE-modified bovine serum albumin (AGEs) or native bovine serum albumin (BSA). The effects of AGEs (0,12.5,25,50,100 and 200 mg·L^-1) on mesangial cells fibronectin and collagen Ⅳ protein contents and the role of PPAR-γ activation in rat mesangial cells ECM accumulation induced by AGEs (100 mg·L^-1) were investigated. Fibronectin, collagen Ⅳ protein contents were detected by ELISA and PPAR-γ mRNA was analyzed by reverse transcriptase-polymerase chain reaction (RT-PCR). Results Mesangial cells expressed PPAR-γ mRNA. Compared with respective BSA groups, AGEs（12.5-200 mg·L^-1）increased fibronectin and collagen Ⅳ protein contents of rat mesangial cells. PPAR-γ activation (1.25-10 mmol·L^-1) could ameliorate collagen Ⅳ protein increasing induced by AGEs （100 mg·L^-1） in mesangial cell medium. Conclusion PPAR-γ activation can ameliorate mesangial cells collagen Ⅳ increasing induced by AGEs.

Objective To study the effects of Gross Saponin of Tribulus Terrestris(GSTT) and Dofetilide on arrhythmias and their mechanisms of antiarrhythmias. Methods Antiarrhythmic effects of GSTT and Dofetilide were observed by arrhythmic models of ligating rat left anterior descending coronary arteries and their antiarrhythmic mechanisms were investigated by isolating cardiac muscle cells in guinea pig and recording delayed outward potassium current(Ikr) and inwardly rectifying potassium current (Ik1) using whole-cell patch clamp technique. Results The number of arrhythmias was decreased significantly by Dofetilide with the dosage of 10 mg·kg^-1，compared with the controls（P<0.01）, however GSTT with the dosage of 20 and 40 mg·kg^-1 did not affect the number of arrhythmias（P>0.05）. Ikr and Ik1 could not be influenced by 50 and 100 mg·L^-1 GSTT, but 5 μmol·L^-1 Dofetilide could significantly reduce Ikr and Ik1 compared with the controls（P<0.01）. Conclusion The antiarrhythmic mechanisms of Dofetilide are probably related with its inhibiting Ikr and Ik1, GSTT has no effects on arrhythmias because it can not influence Ikr and Ik1.

Objective To study the changes of the auditory threshold after two frequencies infrasound exposure in guinea pigs.Methods The electron was inserted into Fallopian′s canal of guinea pigs for a long time during the experiment. The auditory thresholds in 40 awake guinea pigs exposed to infrasound with various frequencies and intensity were observed systematically. Results The permanent threshold shifting (PTS) was caused after 8 Hz and 12 Hz 130 dB infrasound exposure for 30 h and temporary threshold shifting (TTS) was caused after 120 dB infrasourd exposure for 30 h.Conclusion The Ap(N₁) threshold is damaged permanently after 130 dB infrasound exposure.

Objective To study the regulation of T lymphocytes on antigen-specific IgG (sIgG) from B lymphocytes in asthmatic mouse models and clarify the role of T lymphocytes in the pathogenesis of asthma. Methods Firstly asthma mouse models were established. Then the mouse-ventilation-function-testing system was used to select mice which were successfully immunized. T and B lymphocytes were isolated from the mouse spleen aseptically and were cultivated in different combination. Then the supernate was collected and the sIgG in the supernate were tested with enzyme-linked-immunosorbent-assay (ELISA). Results No matter if the specific antigen was involved, sIgG secreted from B cells from asthmatic mice was higher than that of the B cells from control mice (P<0.05 and P<0.01). After regulated by its own T lymphocytes, levels of sIgG secreted by B cells from asthmatic mice decreased significantly (P<0.05). No matter if the T cells from asthmatic mice was involved, the specific antigen couldn′t change the levels of sIgG secreted by B cells from asthmatic and control mice (P>0.05); after interfered by T lymphocytes from asthmatic mice, levels of sIgG produced by B cells from normal control were increased obviously (P<0.05.); after regulated with T lymphocytes from asthmatic mice, levels of sIgG produced by B cells from asthmatic mice was lower than that of the B cells from control mice (P<0.05). With or without specific antigen, after regulated with T cells from asthmatic mice, levels of sIgG produced from B cells of asthmatic mice decreased obviously (P<0.05 and P<0.01). Conclusion sIgG is involved in the mechanism of asthma and T lymphocytes have a key role in regulating the pathogenesis of asthma. And T lymphocyte functional abnormality is one of the important mechanisms of asthma.

Objective To explore the effects of glutamine on hepatocytes apoptosis in rats with obstructive jaundice. Methods 50 male Wistar rats were randomly divided into three groups:control(SL，n=10),obstructive jaundice group (OJ，n=20) and glutamine treatment group (TG，n=20).The hepatocytes apoptosis and the expressions of Bcl-2 and Bax were detected by using TUNEL and immunohistochemistry method. Results The expression of Bcl-2 in rat hepatocytes was increased 2 weeks after treatment in TG group. It expressed positively (+++). The expression of Bax was also decreased compared with the SL and OJ groups in two weeks. It expressed positively (++). Apoptotic index was 15.75±3.68. The difference was significant (P<0.05). Conclusion The glutamine can decrease hepatocytes apoptosis,the expression of Bax, and increase the expression of Bcl-2.

Objective To study the feasibility of placing degradable stent in common bile duct(CBD) and laser welding after CBD exploration (CBDE) in dogs and its clinical values. Methods 30 dogs underwent simulated CBDE were randomly divided into 3 groups:single laser welding (comparative group Ⅰ，n=10), degradable stent inserted in CBD with primary closure (comparative group Ⅱ，n=10), degradable stent inserted in CBD with laser welding (experimental group,n=10). When stent didn′t appeared on the radiograph, the dogs were sacrificed. Evaluating parameters included: pre-operative and post-operative CBD diameters, operation time, bile leakage rate, pathological examinations of liver and CBD tissues 6 weeks after operation. Results The bile leakage rate was significantly higher in comparative groupⅠthan those in experimental group and comparative group Ⅱ. There was an obvious statistical difference in operation time which was longer in comparative group Ⅱ than that in experimental group. Compared with comparative group Ⅱ, the post-operative pathological examination of CBD in experimental group showed that bile mucosa had been repaired entirely, less inflammatory, smoother healing and less scar formation. There were no statistical differences between experimental group and comparative groupⅡ in terms of pathological examination of liver tissue and change of CBD diameter.Conclusion The degradable stent placement after CBDE and the laser welding of CBD is a safe and more effective technique adjunct to CBDE. The method is superior to primary closure.

Objective To observe the time-course and dose-effects of different doses of high voltage prick electrostatic field (HVPEF) on the expressions of CD48 in peritoneal macrophages and CD2 in thymocytes. Methods Whole bodies of mice were treated with HVPEF. Immuno-fluorescent FCM was adopted to detect the changes of CD48 and CD2 expressions. Results CD2 and CD48 expressions increased significantly after treated with lower voltages (10-20 kV)(P<0.01), while higher voltage (35 kV) decreased expressions of CD2 and CD48 (P<0.01); CD2 and CD48 expressions began to increase 2 h and to reach its peak at 4-8 h after treatedith 15 kV (P<0.001); CD2 and CD48 expressions decreased to their minimums 2-4 h after treated with 35 kV, then recovered gradually (P<0.001). Conclusion The different doses of HVPEF might cause different biological effects, which suggests HVPEF has potential immune-regulatory effects and might play an important role in immune modulation.

Objective To observe the repairing effects of a new type of bone graft material constructed by combining calcium phosphate cement (CPC) with bone morphogenetic protein (BMP) on avascular necrosis of femoral head (ANFH) in rabbits; to evaluate the feasibility to use this material to repair ANFH in clinic. Methods The models of bone defect of ANFH were established in the left femoral heads in 24 healthy adult rabbits, of which 12 defects were implanted with CPC/BMP composite, 12 implanted with CPC. The specimens were harvested separately at the end of 3 and 12 weeks after operation (each time 6 rabbits in each group). In order to observe the formation of new bone and the degradation process of the material, a series of examinations were carried out, such as radiograph, histomorphology, transmission electron microscope (TEM). The results in CPC group and CPC/BMP group were compared on the same condition. Results New bone formation in CPC/BMP group was superior to that in CPC group significantly at every stage. In CPC/BMP group, at the 3rd week, new bone and many puerile osteocytes could be found on the surface or inside of CPC, which had been divided into many small parts like islands, with Lamellar bone forming; under TEM, osteoblasts could be found invading CPC, whose rough endoplasmic reticulum (RER) and mitochondrion in plasma increased and swelled. At the 12th week in CPC/BMP group, new bone invaded CPC and enwrapped each other, with trabeculae enlarging; the image of new bone was clear by X-ray examination, with the density of CPC decreased; under TEM, many osteoblasts could be found spreading in CPC, whose RER and mitochondrion in plasma increased and swelled; many capillaries could be found in CPC. The speed of degradation in CPC/BMP group was evidently higher than that in CPC group. Conclusion CPC is an ideal carrier to BMP. The CPC/BMP composite has strong osteoconductibility and osteoinductibility and great potential in ANFH repairing. The speed of degradation of CPC/BMP is evidently higher than that of CPC.

Objective To investigate the effects of Lidocaine on the expressions of HSP70 mRNA and HSP70 protein in cerebral cortex during transient focal cerebral ischemia-reperfusion in rats, and to discuss the possible mechanisms of its neuroprotection. Methods Sixty male Wistar rats weighing 250-300 g were randomly divided into 3 equal groups: sham operation group undergoing sham operation; ischemia/reperfusion (I/R) group undergoing thread embolism of the left middle cerebral artery occlusion (MCAO) to cause focal ischemia for 2 h and then undergoing reperfusion; and lidocaine group undergoing intravenous injection of Lidocaine in doses of 10 mg·kg^-1 10 min before MCAO. At 3, 6, 24, and 72h after reperfusion the rats were decapitated, the expressions of HSP70 mRNA and HSP70 protein in ischemic versus nonischemic cortex were examined using in situ hybridization and immunohistochemistry. At the 24 h after reperfusion, scores of neurological deficit were estimated and changes of histomorphology in ischemic cortex were observed under the light microscope by using HE staining. Results The expressions of HSP70 mRNA and HSP70 protein in ischemic cortexes in the I/R group 3 to 72 h after reperfusion increased significantly, and the peak value of their expression was 161.5±4.8 and 160.9±4.8, respectively, significantly higher than those in the sham operation group (all P<0.01). The expression of HSP70 mRNA appeared early and had a wide distribution in ischemic cerebral hemisphere. Whereas, the expression of HSP70 focused in ischemic cortex. The score of neurological deficit in rats in I/R group was 2.28±0.47, significantly higher than that in the sham operation group (P<0.01). Congestion and edema in ischemic cortex, degeneration and necrosis of neuron were observed in histomorphology in I/R group. Intravenous injection of Lidocaine enhanced significantly the expressions of HSP70 mRNA and HSP70 protein in ischemic cortex 3 to 72 h after reperfusion. The peak values of HSP70 mRNA and HSP70 protein expression in the Lidocaine group were 178.6±5.6 and 176.2±4.7, respectively, significantly higher than those in the I/R group (all P<0.01). The scores of neurological deficit and the damage of histomorphology in the lidocaine group were significantly lower than those in I/R group (all P<0.05). Conclusion Lidocaine can enhance the expressions of HSP70 mRNA and HSP70 protein during focal cerebral ischemia-reperfusion. This may be one of the mechanisms of its neuroprotection.

Objective To study the regulatory effects of polysaccharide Fb from Huangmo (Fb) on immunological function in mice. Methods 40 healthy adult mice were randomly divided into control group (C), CTX immune-suppressed group (CTX), Fb treated group (Fb), Fb combined with CTX treated group (M). The influences of Fb in NK cell activity were determined by the method of NK mediated cytotoxicity test；the multiplicative capacity of lymphocyte was assayed by lymphocyte transformation rate (LTT) of T cell，and the mice spleen T lymphoblast multiplication analytical method was used to measure the IL-2 level. The expressions of MMPs in related immune system were measured by gelatin zymogram electrophoresis. Results Fb increased the NK cell activity, LTT and captivity of IL-2 in immune-suppressed mice（P<0.05），and also promoted the LTT in normal mice （P<0.01）.The weights of spleen and thymus in Fb and M groups were increased, and the body weights of mice in Fb and M groups were increased remarkably than that in CTX group（P<0.01）. The activities of MMP-2（P<0.05）and MMP-9（P>0.05）indicated a decreased tendency in CTX group, the activity of MMP-9 were increased significantly in Fb group and M group（P<0.01）. Conclusion Fb has an activating effect on immune system and may protect and promote the proliferating and repairing of immune system in immune-suppressed mice. Fb can promote the LTT of T cell significantly and may up-regulate some other immune indexes slightly.

Objective To observe the protective effects of XinTongTie (XTT) on acute myocardial infarction in dogs. Methods Twenty four dogs were divided into four groups at random （n=6）：normal control group (NS), low dose of XTT (0.6 g) group, high dose of XTT (1.2 g) group ,and positive control group (NG). Models of acute myocardial infrarction were set up by ligating left anterior descending (LAD) in the anesthetized open-chest dogs.The patch was put on the left pleura near the armpit of every dog respectively. The coronary circulation and cardial oxygen metabolism peremeters were detected 5-360 min after administration. Myocardial infarction size and the changes of the enzymes in serum were determined 360 min after administration. Results The coronary resistance and cardial oxygen consumption were decreased（P<0.05，P<0.01）, the myocardial blood flow was increased （P<0.05，P<0.01）in dogs treated with XTT of 0.6 and 1.2 g. The myocardial infarction size and the activity of serum CK, LDH ,AST were decreased （P<0.05，P<0.01） in acute myocardial infarction dogs treated with XTT of 0.6 and 1.2 g. Conclusion XTT has protective effects on myocardial ischemia through improving the coronary circulation, decreasing cardial oxygen consumption in dogs.

Objective To explore the possibility of achieving the highest resolution by using combination of anion and cation exchange chromatography. Methods Purification results were compared among the differential combination chromatography of anion and cation exchange, anion and cation exchange chromatography of both isocratic elution, and affinity chromatography. Results It was showed by PAGE that the differential combination chromatography of anion and cation exchange had much better purifying ability than anion and cation exchange chromatography of isocratic elution.Conclusion Differential combination chromatography of anion and cation exchange can get good result which can be got under the affinity chromatography conditions in the purification of proteins.

Objective To study the resistance effect of traditional Chinese medicine mixture to the damage of chromosome caused by Bleomycin (BLM) mutagen. Methods Twenty healthy blood donors were divided into four groups:control group,BLM induced group, traditional Chinese medicine mixture group and traditional Chinese medicine mixture anti-induced group. All blood samples were collected at the day of blood culture, hepalean preventing to coagulate. Culture medium were placed into incubator for 72 h after accepted 0.3 mL blood. The cells were harvested and microscope slides were prepared before Giemsa staining. The influence of BLM on the peripheral blood lymphocyte chromosome of the normal body was detected. The antimutagenic effects of the four-herb mixture including Ginseng，Cortex Acanthopanasia radicis，Liquorice，Grifola umbellate as chromosome stabilizer were observed. Results The b/c value in mutagenesis group (0.733±0.275) was obviously higher than that in control group (0.018±0.076)（P<0.01）, the difference of b/c value between mutagenesis group (0.733±0.275) and traditional Chinese medicine mixture anti-induced group (0.566±0.761) was significant（P<0.01）. Conclusion BLM can increase the fracture rate of the peripheral blood lymphocyte chromosomes in normal body obviously, the mixture including four traditional Chinese medicine can inhibit the fracture rate of chromo some induced by BLM obviously. It suggests that the mixture including four traditional Chinese medicine can improve the genome stability of the organism as chromosome stabilizer.

Objective To study the effects of 20(S)-ginsenoside Rg₃ (SPG-Rg₃) on growth of sarcoma 180 and life of mice with sarcoma 180.Methods In vitro,stain exclusion experiment was used to observe the effect of SPG-Rg₃ on sarcoma 180 cells. In vivo, the mice models of sarcoma 180 were built and divided into 6 groups: 3，1， and 0.3 mg·kg^-1 SPG-Rg₃ groups, CTX group, control group, and SPG-Rg₃ combined with CTX group. Twenty-four hours after inoculating sarcoma cells, mice were administrated by abdominal cavity and killed at the 10th day. The body and neoplasm weights were detected, and the growth suppression rate of tumor and CDI were calculated. The model of ascites sarcoma 180 was also built and CTX was replaced by 5-FU. The method of diviving groups was same as before. Mice were administrated for 5 d and after that, the life of mice with sarcoma 180 was recorded and life elongation term was calculated. Results In vitro, SPG-Rg₃ inhibited the growth of sarcoma 180 cells and the IC⁵⁰ value was (10.49±2.96)mg·L^-1. In vivo, SPG-Rg₃ also inhibited the growth of sarcoma 180 and the inhibitory effect of SPG-Rg₃ combined with CTX was superior to those of SPG-Rg₃ or CTX. SPG-Rg₃ and 5-FU also prolonged life of mice and the rate of life elongation in SPG-Rg₃ combined with 5-FU group was higher than that in 5-FU group. Conclusion SPG-Rg₃ can inhibite the growth of sarcoma 180 and prolong the life of mice with sarcoma 180. Foremore, SPG-Rg₃ can reinforce the inhibitory effects of CTX and 5-FU on tumor growth.

Objective To study the effects of Rhodosin and deer serum preparation on aging skin. Methods 12 guinea pigs of 20 months with 3 cm×3 cm hair sheared from their backs were divided into control group and experimental group. Rhodosin and deer serum preparation was besmeared on the skin of guinea pigs in experimental group, and cosmetic matrix was besmeared on the skin of animals in control group for 60 d. The distribution of type Ⅲ collagen was observed with Sirius red staining and HE staining. The fission index of the cells of stratum basale, the thickness proportion of stratum corneum to epidermis, the volume density of the collagen fibers and small blood vessel in dermis, the number of fibroblasts in aging skin were meassured with technique of sterology. Results Compared with the control group, the distribution of type Ⅲ collagen in papillary layer, the fission index of the cells of stratum basale, the number of fibroblasts in papillary layer and the volume density of small blood vessel were increased in the experimental group (P<0.01); the volume density of the collagen fibers in dermis and the thickness proportion of stratum corneum to epidermis were reduced in the experimental group (P<0.01 or P<0.05). Conclusion Rhodosin and deer serum preparation can defer the aging on skin.

Objective To study the inhibitory effects of cyclooxygenase-2-selective inhibitor on angiogenesis postkeratoplasty. Methods The rat keratoplasty models between SD and Wistar rats were made. All the rats were taken orally cyclooxygenase-1-selective inhibitor,cyclooxygenase-2-selective inhibitor，non selective-cyclooxygenase inhibitor, and physiological saline separately. The rejection values (RV) according to edema,opacity and new vessel of the cornea graft were measured. The new vessel response area was calculated by observing the new vessel of cornea. Results The RV values of cyclooxygenase-1-selective inhibitor and control groups were more than 5 at the 7 day after operation. Rejection had been found in these two groups while it hadn′t in other two groups. The new vessel of cyclooxygenase-2-selective inhibitor group and cyclooxygenase-1-selective inhibitor group at the 7 day could be detected by slit lamp on the cornea bed. The new vessel of other groups approached the cornea graft. The pathological section examination had the same conclusion. The value of RV and the new vessel response area of the groups in which the rats took cyclooxygenase-2-selective inhibitor and non selective-cyclooxygenase inhibitor were less than those of other groups. Conclusion Cyclooxygenase-2-selective inhibitor can inhibit angiogenesis postkeratoplasty.

Objective To observe the effects of Astragalus Membranaceus injection(AMI) on coxsackievirus B₃ (CVB₃) murine viral myocarditis.Methods 70 BALB/C mice were divided randomly into control group,model group,AMI 90,60,18 g·kg^-1 groups (high,middle,low dose groups) and positive group. Viral myocarditis models induced by intraperitoneal inoculation with CVB₃ solution were developed and were treated by high,middle,low doses of AMI respectively. Nine days after administration,the survival rates and the levels of serum AST,LDH,SOD and MDA,the changes in ECG and myocardial tissues were observed. Results The survival rate of the mice was increased and the incidence of abnormal ECG at acute stage was significantly reduced when AMI was used, compared with normal group (P<0.05,P<0.01). The levels of serum AST,LDH,MDA were significantly decreased in AMI treated group,and the activities of SOD were increased especially in middle and low doses of AMI groups (P<0.05,P<0.01).AMI reduced obviously the degreee of lesion in mice with myocarditis. Conclusion AMI exert antagonistic effects on CVB₃ murine myocarditis.

Objective To investigate the mechanism of temporal lobe epilepsy through observing ultrastructural changes of CA1 region in hippocampus in temporal lobe epileptic male rats. Methods Temporal lobe epileptic typical models were established in Wistar male rats kindled by kainic acid microinjected into right Nucleus amygdalae. Ultrastructural observations on nerve cells and synapses in CA1 region were performed, as well as quantitative analysis on their active parameters on 14 d, 21 d after kainic acid injection in five rats, respectively. Results There existed nerve cells structural damage in CA1 region in temporal lobe epileptic rats, synaptic parameters including synaptic density, synaptic active area, synaptic interface curvature, surface ratio, synaptic vesicles in epileptic group were significantly decreased than those in sham group（P<0.05）. The number or volum of mitochondria in astrocytes in epileptic group were significantly increased than those in sham group（P<0.05）. There existed long-term ultrastructural and active paramete changes in nerve cells and synapses.Conclusion There are structural and functional injuries in nerve cells and synapses in CA1 region in temporal lobe epileptic male rats.

Objective To optimize the expression condition and increase the output of recombinant HspA-UreB fusion protein of Helicobacter pylori. Methods The recombinant HspA-UreB fusion protein of Helicobacter pylori cloned in E.coli BL21 was expressed in different revulsive concentrations (0.3, 1.0 and 2.0 mmol·L^-1) and culture medium (LB, SOC) at different temperatures (25℃, 37℃) for different time, the offsprings expressed in E.coli BL21 (pET-HU27) were broken up with ultrasonic, and Western blotting test was used to detect the offsprings. Results The output of this fusion protein was highest in LB culture medium in 0.3 mmol·L^-1 IPTG concentration at 37℃ for 5 h, and the percentage of soluble fusion protein was highest in SOC culture medium in 1.0 mmol·L^-1 IPTG concentration at 25℃ for 8 h, and the offspring could be recognized specifically by mouse serum. Conclusion HspA-UreB fusion protein of Helicobacter pylori is expressed with high efficiency and with good immunoactivity, and which will help to develop gene recombinant vaccine against Helicobacter pylori infection.

Objective To investigate the preliminary effect of oxymatrine　(OMT) on inflammation and its mechanism. Methods Fifty mice were randomly divided into 5 groups: normal saline group, dexamethasone group and OMT groups with different doses (70,140,and 210 mg·kg^-1). The extent of ear swelling was observed in inflammation model caused by dimethyl benzene in mice. Forty-eight Wistar rats were randomly divided into 6 groups:normal saline group, dexamethasone group and OMT groups with different doses (50，100,and 150 mg·kg^-1),L-NAME+OMT 100 mg·kg^-1 group. Paw edema volume induced by dextran were observed in rats. Results The extents of ear swelling were decreased in dose-dependent manner after intraperitoneal injection of OMT 140 and 210 mg·kg^-1 in mice compared with control (P<0.05 or P<0.01). Paw edema volumes were reduced after intraperitoneal injection of OMT 100 and 150 mg·kg^-1 in rats compared with control (P<0.05). L-NAME enhanced the effect of anti-inflammation of OMT. Conclusion OMT can reduce the extent of ear swelling induced by dimethylbenzene in mice and the paw edema volume caused by dextran in rats.Its anti-inflammatory effect may be not fully mediated by NO-cGMP pathway.

Objective To observe the effects of instability of cervical vertebral column on the disturbance of the special memory, intelligence, blood urine in mice. Methods 300 mice of 6-month-old were divided equally into an experimental group and a control group. The experimental mice were injected with 15 μL of 30% lactic acid (once every week, 3 weeks continued) in the 1st and 2nd cervical articulation. The control mice were injected with 15 μL of 30% lactic acid (once every week, 3 weeks continued) in the subcuteous tissue which was in the area at upper the gluteobiceps muscle. The memory and intelligence of mice were examined with the conditional avoidance test, float harbor test and white paper test. Results The experimental mice showed special memory and intelligence deficiencies (P<0.01) and the decay of health, change of hematopoietic function and positve urinary protein (P<0.01). Conclusion Instability of cervical vertebral column has correlation with dementia, meanwhile,the stimulating degree to organism is bigger.

Objective To observe the configuration and traits of valva mitralis. The circumference of the margin of left atrioventricular orifice, and the attached part of cuspis anterior and cuspis posterior were measured; the area of the orifice, cuspis anterior and cuspis posterior were calculated, in order to clarify the relationships of them and provide data for vavular disease repair operation and vavle transplantation. Methods The measurement and macroscopic investigation were performed on the hearts from 24 embalmed cadaver specimens. The most part of the left atrium that was above the orifice were removed. Then the circumference and area of the left atrioventricular orifice were measured. The left ventricle along the long axis of heart from the posteriomedial commissural part was dissected. The chordae tendinease was cut off. The ventricle was opened and made plain. Then the length of attached margins and area of the valves were measured. Results The area of the cuspis anterior was （6.487±1.238）cm² while it was （5.363±1.461）cm² of the orifice and （4.036±1.658）cm² of the cuspis posterior. The area of the orifice was about 0.524±0.121 of that of the total valves. The circumference of the orifice was （8.175±2.116）cm, and the margin of the attached part of cuspid anterior was （4.474±1.169） cm while it was （3.701±0.954）cm of the cuspis. Conclusion The data accumulated anatomically is better than that of common radiology and ultrasonic cardiography indirectly. The area of cuspis anterior is larger than that of the cuspis posterior, and the valva mitralis has morphologic potential and reservation. It has more clinical significance in vavle repair surgery and valve transplantation.

Objective To inform the effects of Se and vitamin E on the anabolism of type Ⅰ collagen in cultured cardiac fibroblasts. Methods Cardiac fibroblasts were cultured using culture solution containing different concentrations of Se and vitamin E (VE), Se+VE for 48 h, then the contents of hydroxyproline and type Ⅰ collagen in cultured solution were detected. Results ①Compared with control and NE groups, the contents of hydroxyproline could be significantly decresed in all the three Se group (low 0.05 mg·L^-1, middle 0.1 mg·L^-1, high 0.2 mg·L^-1) (P<0.05), and this effect became apparent when the content of Se was 0.2 mg·L^-1 (56.13±1.31, P<0.05); ②Compared with control and NE groups, the contents of hydroxyproline were decreased significantly in all the three VE groups (1, 10, and 100 mg·L^-1), especially in 1 and 100 mg·L^-1 groups（P<0.05）; ③Both the contents of hydroxyproline and type Ⅰ collagen were decreased when Se (0.05 mg·L^-1) and VE (10 mg·L^-1) applied simultaneously. Conclusion The anabolism of type Ⅰ collagen in cultured cardiac fibroblasts can be inhibited by Se and VE.

Objective To investigates the influences of focal cerebral ischemia on CRH，NGF and HPA in hypothalamus of rats. Methods Thirty-two senile Wistar rats including fourteen female and fourteen male rats were divided into seven groups at random and there were four rats per group. One group was control group, the other six groups were operation groups. Operation groups consisted of ischemia 15 min group and ischemia 15 min groups in which rats were reperfused for 1，6 h and 1，2，4，9 d. The expressions of CRH and NGF in hypothalamus of focal cerebral ischemia senile rats were observed dynamically with the immunohistochemical method. Results There was a little expressions of CRH and NGF in hypothalamus in control group, the expressions of CRH and NGF in hypothalamus didn′t change during 1 h after ischemia reperfusion in operation groups, then the expressions of CRH and NGF in hypothalamus disappeared transiently at 6 h after ischemia reperfusion, but after that the expressions of CRH and NGF in hypothalamus increased obviously at 2 d after ischemia reperfusion. Conclusion The activity of CRH secretory cells in hypothalamus is not immediately in the strength phase after cerebral ischemia; the activity strength effect of CRH secretory cells in hypothalamus exists in the later stage after cerebral ischemia; NGF is possibly important to the activity of HPA.

Objective To compare the toxicities between gentamicin and its metabolite, and study the effects of Ca²⁺and Mg²⁺ on the toxicity gentamicin. Methods ①The spiral organ of corti of 10 mice (20 ears) were taken, and divided into 2 groups randomly (n=10), cultured in 0.1 mol·L^-1 gentamicin and the metabolite of 0.1 mol·L^-1 gentamicin for 48h, then the number of hair cells remain was recorded;②The spiral organ of corti of 10 mice were taken (20 ears), and divided into 2 groups randomly (n=10), 3% CaCl₂ and 0.1 mol·L^-1 GM were mixed in the culture medium and the number of hair cells remain in the two medium mentioned above were recorded; ③The spiral organ of corti of 10 mice were taken (20 ears), divided into 2 groups randomly (n=10), 3% MgSO₄ to 0.1 mol·L^-1 GM were mixed in the culture medium, the number of hair cells remain in the two medium mentioned above were recorded. Results It was not significant difference between 0.1 mol·L^-1 GM and the metabolite of 0.1 mol·L^-1 gentamicin (P＞0.05) ; but when 3% CaCl₂ or MgSO₄ were mixed in the culture medium, the number of the survived hair cells was observably increased (P＜0.01). Conclusion The toxicity of GM to the cochlea hair cells has nothing to do with its metabolite; Ca²⁺and Mg²⁺ may act as antagonist to the toxicity of GM in cochlea hair cells.

Objective To explore the possibility of establishment of the animal model of West syndrome. Methods Experiment was performed on 12-day-old Wistar strains (n=64). 32 among them were randomly divided into N-methyl-D- Aspartate (NMDA) treated group（n=16）and control group（n=16）, and had an intraperitoneal injection of NMDA or NS respectively for 14 d in succession. The behavioral manifestations and/or the seizures induced by NMDA in each group were observed and the incidence, latencies, pattern of motor seizures were noted. The changes of ECoG were observed in the rest of Wistar strains (n=32) which were divided into NMDA treated group (n=16) and control group（n=16）. Results The control group had no seizures. NMDA-induced automatisms and emprosthotonus in (12-13)-day-old rats were elicited while automatisms were only elicited by NMDA from 14-day-old rats to 25-day-old rats without emprosthotonus. There were no obvious epileptiform discharge in control rats compared with the NMDA-treated rats which had the characteristic changes of shape waves and spike waves concurrently during automatisms and emprosthonotonic seizures. Conclusion Although the unique seizures with age-specificity induced by NMDA in developing rats can not fulfills all criteria of the West syndrome model, it partially resembles West syndrome in childhood on the aspects of behavioral manifestations and ECoG changes.

Objective To investigate the association between schizophrenia and the polymorphism of 5-hydroxytryptamine receptor 2A (HTR2A) gene. Methods The PCR-based restriction fragment length polymorphism (RFLP) technique was used to detect a single nucleotide polymorphism（SNP）, rs6313 locus, at the HTR2A gene in 54 Chinese Han family trios. Results The genotypic distribution of HTR2A was not deviated from the Hardy-Weinberg equilibrium; the haplotype relative risk test (HRR) did not show a genetic association between HTR2A and schizophrenia (χ²=1.852, df=1, P=0.174); transmission disequilibrium test (TDT) did not show significantly biased transmission from parents to affected offsprings (χ²=1.786, df=1, P=0.181); there were correlations between allele frequencies and two clinical symptoms of schizophrenia, bizarre behaviors (χ²=5.576, df=1, P=0.018) and abulia (χ²=8.322, df=2, P=0.016).Conclusion The polymorphism of rs6313 locus of HTR2A gene may not be associated with schizophrenia although it may be associated with certain psychotic symptoms.

Objective To investigate the genetic association of metallothionein (MT) 2A gene locus in chromosome 16 with susceptibility to type 2 diabetes mellitus (T2DM) in Chinese Han people. Methods The techniques of PCR and restriction fragment polymorphism(RFLP)were used to examine single nucleotide polymorphism(SNP)of MT2A gene in 129 patients with T2DM（case group） and 115 unrelated healthy individuals(control group). Results The distributions of genotype frequencies of rs10636 were not deviated from the Hardy-Weinberg equilibrium; there were no remarkable differences of the frequency distributions of GG,GC and CC of MT2A between cases and controls (χ²=0.202,P>0.05),and the frequency distributions of G and C had no markedly differences between cases and controls (χ²=0.136,P>0.05).Conclusion The rs10636 of MT2A gene may not be associated with T2DM.

Objective To study the expression of a novel apoptosis inhibitor，survivin，in laryngeal carcinoma and its relationships with apoptosis and clinical pathologic parameters. Methods The expressions of survivin protein in Hep2 cell line, periodontal ligament cells (PDLC),40 laryngeal carcinoma and 8 normal laryngeal tissue were detected by immunohistochemical staining. The expression of survivin mRNA was detected by RT-PCR. DNA in situ end labelling technique was used to detect apoptosis index (AI). Results Survivin protein expressed in Hep2 cell line and 23 of 40 (57.5%) cases of laryngeal carcinoma. In contrast， it didn′t express in PDLC and normal laryngeal tissues. The positive percentage of survivin protein in tumor grade Ⅰ laryngeal carcinoma (33.3%) was strikingly lower than that of grade Ⅱ and Ⅲ laryngeal carcinoma (72.7%，P<0.05). The positive percentage of survivin protein in clinical stage Ⅲ and Ⅳ laryngeal carcinoma (76.2%) was strikingly higher than that of grade Ⅰand Ⅱ laryngeal carcinoma　(36.8%， P<0.05). The positive percentage of survivin protein in laryngeal carcinoma with lymph node metastasis (100%) was strikingly higher than that of laryngeal carcinomas without lymph node metastasis (48.5%,P<0.05,). AI of survivin protein positive cases (2.93±1.20) was lower than that of negative cases（4.05±1.42,P<0.05). Conclusion The results indicate that apoptosis inhibition caused by survivin protein may play a important role in the origin and progression of laryngeal carcinoma.

Objective To study the expressions of NF-κBp65 and VEGF in brain glioma and its infiltrative tissues and their significances.Methods The expressions of NF-κBp65 and VEGF were detected in 53 cases of brain gliomas, 26 cases of infiltrative tissues and 5 cases of normal tissues by immunohistochemical technique. Results The expressions of NF-κBp65 and VEGF were correlated with the differentiation of brain glioma （P<0.05）. In infiltrative tissues of high malignant glioma, the expressions of NF-κBp65 and VEGF were higher than those in low malignant gliomas. The expression of NF-κBp65 in brain gliomas was positively related to the expression of VEGF （r=0.695 2，P<0.05）. Conclusion NF-κBp65 as a oncogenic protein related to brain gliomas may have positive regulation on VEGF and promote the tumor angiogenesis and deterioration.

Objective To study the CT, MRI performance and invasion paths outside paranasal sinus malignant tumor. Methods The data of CT and MRI of 31 patients with paranasal sinuses malignant tumor were analyzed, tumor′s invasion scope and invasion paths outside according to the CT, MRI signs and symptoms. Results Paranasal sinus malignant tumor root mainly in maxillary sinus 17　cases (54.8%), then ethmoid sinus 10 cases (32.3%), and frontal sinus 4 cases (12.9%). The CT and MRI results showed soft tissue disease focus was found and its boundary was not clear, enhancement CT and MRI scanning showed 8 and 11 cases had low and middle degrees enhancement,respectively. The pathological changes in 22 cases often be outside the nasal cavity and paranasal sinus but infringed upon the pterygopalatine fossa, orbit. Conclusion CT and MRI can display exactly tumor′s primary focus and its invasion outside the paranasal sinus and judge its invasion path.

Objective To evaluate the corrosion resistance of Ni-Cr alloy,Ti-6Al-4V alloy and Au-Pt alloy in artificial saliva. Methods Nine speciments of Ni-Cr alloy,Ti-6Al-4V alloy and Au-Pt alloy were made and they were divided into three groups(n=3). Then anodic polarization test was performed to measure the Ecorr and Anodic potentiondynamic polarization curves of each group. Surface changes of the alloys were observed by SEM. The products of corrosion were analyzed by X-ray diffractiometer. Results Ecorr of Ni-Cr alloy, Ti-6Al-4V alloy and Au-Pt alloy were -115,-24 and +61 mV. Anodic potentiondynamic polarization curves showed at the certain potential(<600 mV),the Ti-6Al-4V and Au-Pt alloys had lower current density than Ni-Cr alloy. SEM results showed that the corrosive marks on the surface of Ni-Cr alloy were evident and that of Ti-6Al-4V were not very evident. There were not corrosive marks on the surface of Au-Pt alloy. X-ray diffractogram showed that the corrosive product was mainly Cr chemical compound on the surface of Ni-Cr alloy and Ti chemical compound on the surface of Ti-6Al-4V alloy. The chemical compound in Au-Pt alloy was not detected. Conclusion In artificial saliva,the corrosion resistance of Au-Pt alloy and Ti-6Al-4V alloy are superior to that of Ni-Cr alloy.

Objective To study the effects of Modified Papanicolaou stain,Wright-Giemisa stain and Coomassie brilliant blue stain on the rate of morphogically normal sperm and the rate of acrosome intactness. ethods Semen smears from 55 male infertility patients were stained according to Wright-Giemisa, Modified Papanicolaou and Coomassie brilliant blue methods. The rate of morphologically normal sperm and the rate of acrosome intactness were evaluated. Results ①No significant differences were found in the percentage of morphologically normal sperm between the three staining methods (P>0.05). Comparing with Modified Papanicolaou stain and Coomassie brilliant blue stain, the rate of Neck/Midpiece defect sperm ,as well as tail defect sperm , significantly decreased after Wright-Giemisa stain（P<0.05）. ②No significant difference was observed in the rate of sperm with type I acrosome and the rate of sperm with type Ⅳ acrosome between the three staining methods (P>0.05). However, the rate of sperm with type Ⅱ or Ⅲ acrosme was higher using Modified Papanicolaou stain than using the other two stains（P<0.05）. ③The significantly positive correlationship was found in the rate of normal morphologically sperm (r=0.535 P<0.01，r=0.601,P<0.01 , r=0.329,P<0.05), as well as in the rate of acrosome intactness （r=0.604, P<0.01，r=0.682,P<0.01, r=0.446,P<0.01）,among the three staining methods. Conclusion The three staining methods stain would be reliable and applicable to detect the percentage of human morphologically normal sperm and the rate of acrosome intactness. Among the three staining methods, Modified Papanicolaou stain would be better to be used in examing the percentage of human morphologically normal sperm, and Wright-Giemisa stain would be more suitable for detecting the rate of acrosome intactness.

Objective To study the effects of Percoll gradient centrifugation and swimming up technique on the sperm viability, the hypoosmotic swelling rate of human spermatozoa tail and the percentage of morphologically normal spermatozoa. Methods Semen samples were collected from 12 male infertility patients. Sperm selection was performed respectively using Percoll gradient centrifugation and swimming up technique. Sperm viabilities and the hypoosmotic swelling rates of human spermatozoa tail before and after sperm selection were analyzed by Eosin stain. Sperm morphological changes before and after sperm selection were evaluated by Modified Papanicolaou stain. Results ①The sperm viability and the hypoosmotic swelling rate of human spermatozoa tail were significantly increased after Percoll gradient centrifugation and swimming up （P<0.01）. No significant difference was found in comparison of sperm viability and the hypoosmotic swelling rate of human spermatozoa tail between the two selection methods（P>0.05）. The rate of sperm with type B-G tail swelling was significantly improved after Percoll and swimming up（P<0.01）, However, there was no significant difference of type B-G tail swelling rate between the two selection methods. ②The rate of morphologically normal sperm significantly was increased after Percoll and swimming up, Meanwhile,the rate of normal morphological sperm after swimming up was higher than that after percoll（P<0.05）. The rate of Tapered sperm, Irregularity sperm and Neck/Midpiece defect sperm after swimming up were significantly decreased（P<0.05）. The rate of Tapered sperm,Irregularity sperm, and Tail defect sperm after Percoll were significantly decreased. However, the rate of Irregularity sperm, Neck/Midpiece defect sperm and tail defect sperm after swimming up were significantly lower than those after Percoll (P<0.05).Conclusion After sperm selection using Percoll gradient centrifugation and swimming up technique, the sperm viability, hypoosmotic swelling rate of human spermatozoa tail and rate of morphologically normal spermatozoa are obviously improved. These two sperm selection methods may be widely applied in reproductive medicine. Swimming up technique is better than Percoll gradient centrifugation.

Objective To study the sensitivity of p21-transfected human gastric cancer cell line (BGC) to chemotherapeutic drugs. Methods MTT assay was used to test the inhibitory effects of Fluorouracilum (5-Fu )，Adriamycin (ADM)， Vincristin (VCR)，Cytoxan (CTX)，Pingyanmycin (PYM) and Teniposide (VM-26) on p21-transfected human gastric cancer cells. The sensitivity of p21-transfected human gastric cancer cells to chemotherapeutic drugs was observed for 72 h. Results p21-transfected gastric cancer cells were the most sensitive to 5-Fu, then ADM，VCR，CTX，PYM and VM-26. The inhibitory rate was 77.9%，62.7%，61.1%，59.3%，58.2% and 45.9%，respectively. The difference of chemosensitivity compared with control was significant (P<0.05). The sensitivity increased with prolongation of exposure time. Conclusion MTT assay is used to test the sensitivity of the gastric cancer cells transfected with p21 gene to chemotherapeutic drugs. Pretreatment determination of chemosensitivity can not only screen out sensitive drugs for individual tumor, but also avoid toxicity of ineffectual drugs to immune cells. p21 gene may increase the sensitivity of cancer cells to 5-Fu.

Objective To explore the effects of E-cadherin (E-cd) gene mutation and its abnormal protein expression on the metastasis of laryngeal cancer. Methods Mutations of E-cd gene exon 5，7 and E-cd protein expressions in 64 cases of laryngeal cancer were detected with the methods of PCR-SSCP-DNA Sequencing and immunohistochemistry. Results E-cd mutation frequency was 17.2%（11/64） in the laryngeal cancer tissues. The E-cd gene mutation was apparently associated with clinical stage and lymph node metastasis (P<0.05). And low expression of E-cd protein was found in laryngeal cancer with metastasis (P<0.05). Conclusion The E-cd gene mutation and abnormal expression of E-cd protein could play a role in the lymph node metastasis of laryngeal cancer by inhibiting the tumor cell adhesiveness and increasing its invasiveness.

Objective To research the association of β-fibrinogen bcl-I polymorphisms and plasma fibrinogen level with the pathogenesis of ischemic heart disease. Methods 57 patients with myocardical infarction and 50 healthy controls were enrolled in this trail. The polymorphisms of β-fibrinogen bcl-I gene was detected by polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLP). Plasma fibrinogen levels were assayed by tissue thromboplastin. Results Two allele, B1 and B2, were found at 2 500 bp and 1 100 bp+1 400 bp，respectively. The plasma fibrinogen level, （3.28±0.94）g·L^-1, in patients with allele B1 was significantly higher than that in the healthy control subjects,（2.74±0.54）g·L^-1 （P<0.05）, as well as with allele B2（3.97±1.02）g·L^-1 vs（3.25±0.61）g·L^-1. The plasma fibrinogen level in the samples of genotype B2 was higher than thatin samples of B1（P<0.05）. The frequency of genotype B2 allele was higher in cases with myocardical infarction than that in healthy controls（P<0.05）. Conclusion There is significant association between the fibrinogen plasma level and myocardical infarction. β-fibrinogen bcl-I gene polymorphism is associated with plasma level of fibrinogen and myocardical infarction. These suggest that β-fibrinogen bcl-I gene polymorphism may be a risk factor and a susceptible predictor of ischemic heart disease.

Objective To study the drug sensitivity of six kinds of chemotherapeutic drugs for human non-small cell lung cancer(NSCLC), and their relationships with the expressions of two drug-resistance factors p53 antibody and GST-π. Methods Fifty-eight NSCLC specimens obtained by operation were used for primary culture. Then the sensitivities of the cells to six kinds of chemotherapeutic drugs were detected by MTT assay. The serum GST-π and p53 antibody in the patients were tested by ELISA at the same time. Results The effective rates of the six kinds of drugs were Cisplatin (DDP) 41.4%, Vicristine (VCR) 24.1%,Etoposide (VP16) 22.4%, Hydroxyl-camptothecin (HCPT) 20.7%,Adriamycin (ADM) 17.2%,Cyclophosphamide (CTX) 17.2%,respectively. The patients had different multi-drug resistance when they had different positive expressions of p53 antibody and GST-π. Conclusion It is possible and necessary to test the drug sensitivity of lung cancer specimens obtained from operation by MTT method, and the results are very useful for the clinic chemotherapy after operation, especially when p53 antibody and/or GST-π positively express in the tissues.

Objective To measure the levels of serum vascular cell adhesion molecule-1(VACM-1),intercellular adhesion molecule-1 (ICAM-1) and C-reactive protein（CRP） in patients with acute coronary syndrome (ACS), and to explore their significances in diagnosis and treatment of coronary heart disease. Methods The levels of serum VACM-1,ICAM-1 and CRP were measured in 50 patients with ACS undergoing coronary angiography as well as 50 controls. Results The levels of serum VACM-1,ICAM-1 and CRP in patients with ACS were higher than those in controls (701±54.6 μg·L^-1vs 556±42.2 μg·L^-1,389±23.7 μg·L^-1 vs 271±34.6 μg·L^-1,7.05±3.13 mg·L^-1 vs 4.22±1.41 mg·L^-1)(P<0.01)；the levels of VACM-1 and ICAM-1 between acute myocardial infraction（AMI） and unstable angina（UA） patients were not difference (699.12±62.77 μg·L^-1 vs 706.57±53.65 μg·L^-1,390.39±42.34 μg·L^-1 vs 372.63±32.59 μg·L^-1)(P>0.05)；CRP content in AMI patients was higher than that in UA patients （8.06±2.78 mg·L^-1 vs 6.33±2.01 mg·L^-1）(P<0.05). Conclusion The stenosis and block caused by atherosclerosis plaque accompany the increase of VACM-1,ICAM-1 and CRP, and there is a relation with condition of the disease. The excess expression of adhesion molecules may be an important factor of atherosclerosis. Adhesion molecule might become a testindex to monitor the occurrence and development of atherosclerosis.

Objective To study the curative effect of arthroscopic debridement (AS) and intra-articular sodium hyaluronate injection for patients suffering from knee osteoarthritis (KOA) and explore their influences on the levels of interleukin-1 (IL-1) and nitric oxide(NO) in serum and synovial fluid. Methods 69 patients with grade Ⅰ to grade Ⅲ KOA were randomly divided into AS group (n=32) and SHI group (n=37). All of the cases were estimated by Lysholm scoring scale pre-intervetion and 1 year post-intervetion,and then serum and synovial fluid were collected to detect the levels of IL-1 and NO before and after therapy,respectively, with Nitrate reductase and enzyme-linked immuneoabsordent assay (ELISA). Results Lysholm scores in the two groups were significantly different between pre-operation and post-operation (P<0.05),and the rate of satisfaction one year post-operation in AS group was 78.0%, superior to 58.3% in SHI group.In addition,before therapy,levels of IL-1 and NO in synovial fluid were obviously higher than those in serum in KOA patients (P<0.05). There were significant differences of levels of IL-1 and NO in serum and synovial fluid between pre-operation and post-operation (P<0.05). Conclusion Both AS and SHI can increase the Lysholm score in KOA patlents,decrease the levels of IL-1 and NO in serum and synovial fluid. AS is superior to SHI in functional improvement.