Objective To provide fundamental data for developing an effective immunization strategy by different types of recombinant vaccines to protect HIV-2 infection. Methods Using prime-boost immunization strategy，BALB/c mice were inoculated muscularly with HIV-2 gp105 recombinant DNA vaccine and fowlpox virus. The numbers of CD4⁺ and CD8⁺ subgroup of spleen T lymphocyte, the specific killing activities of spleen CTL and the titers of serum antibodies of the immunized mice were detected. Results HIV-2 specific target-killing activity of spleen CTL and serum HIV-2 specific antibody level were higher in combined immunization group than those in single immunization groups (P<0.05). Conclusion The strategy of prime-boost may stimulate stronger specific celluar and humoral immune responses in mice.

Objective To investigate pathological characteristics of peripheral nerves and muscles and the relationship between these changes and neuroelectrophysiological changes after methamidophos poisoning in hens. Methods Fifty Leghorn hens were divided into 5 groups at random（n=10）. Experimental groups were given methamidophos through a cannula by mouth and were injected atropine sulfate to live through urgent period. The per oneal nerves, gastrocnemius muscles and sciatic nerves were taken at the end of first, second ,third, fouth and eighth week. Morphological study was carried out. Results Mild myelinated fiber abnormalities were found in peroneal nerves at end of second week in experimental group. Nerve fiber degenerat ion, heaven-stained myelin sheath and axonal swelling were occurred at end of the third and fourth week. Fiber necrosis in the gastrocnemius muscles were seen at the third and fourth week. Low amplitude was shown in neuroelectrophysiological examination of sciatic nerves at first week after poisoning. Till the eighth week, motor nerve conduction velocity and terminal latent period renewed to normal. Conclusion Earlier pathological changes can be found by neuroel ectrophysiological examination, while neuropathological changes of peripheral n erves and muscles can be used to judge prognosis.

Objective To evaluate the influences of high hydrostatic pressure (HHP) on the virulence and antigenicity of Coxsackie virus B₃ (CB₃V) , to explore a new physiological method to develop an active vaccine against Coxsackie virus. Methods The phagocytosis of the macrophages in mice to the CB₃V and the TCID₅₀ of the CB₃V before and after treated by HHP were examined. Results TCID₅₀ increased from 10^-5 to 10^-4 wh en treated by 680 MPa; viruses were inactivated at 700 MPa. Viruses were als o inactivated by the pressure less than 700 MPa, but needed prolonged time. Th e phagocytic function, antibody and T lymphocyte transformation rate in experime ntal mice (heated, attenuated, and inactive viruses) exhibited a significant dif ference compared with control mice (P<0.05). Conclusion CB₃V has a strong resistance to HHP. The inactivat ed pressure is about 700 MPa, however, the antigenicity of the virus is not infl uenced by the same value of HHP.

Objective To construct the prokaryotic expression vector of HSP65-T cell epitopes of MOMP of C.trachomatis（Ct）fusion protein(called H-ctm1), express and purify H-ctm1. Methods The HSP65 and gene sequence of T cell epitopes of MOMP of Ct (ctm1) were obtained by PCR method and cloned into pMD18-T vector respectively. The T vector was digested by endonuclease for releasing the fragment of H SP65 or ctm1 gene. The gene of HSP65 and ctm1 were cloned into pET28a plasmid su ccessively to construct the prokaryotic expression vector (pET28a-H-ctm1), whi ch recombined the gene of HSP65 and ctm1. The E.coli BL21(DE3) transformed with pET28a-H-ctm1 were induced by IPTE for expression of H-ctm1 fusion protein. The protein was purified by Ni²⁺ affinity chromatography. Results The prokaryotic expression vector, pET28a-H-ctm1, was constructed successfully. The fusion protein of H-ctm1 was expressed in E.coli BL21(DE3). The purity of fusion protein was 98% after purified by Ni²⁺ affinity chromatography. Conclusion The prokaryotic expression vector of pET28a-H-ctm1 has been constructed, and the fusion protein with biological activity has been successfully expressed and purified.

Objective To study the effect of fluoride on the expressions of collagen typeⅠ(ColⅠ) and collagen type Ⅲ (Col Ⅲ) in the myocardium of rats.Methods The rat model was established by administration of different concentration o f sodium fluoride. The myocardium of rats were stained by picrosirium red，observed by polarization microscopy，analyzed by image analysis system. The expressions of ColⅠand Col Ⅲ in myocardium of rats were determined by immunohistochemistry method.Results The percentages of ColⅠand Col Ⅲ area in the myocardium of rats treated with different concentration of sodium fluoride were a l ittle higher than those of control group.No significant difference was seen among different groups of the expressions of ColⅠand Col Ⅲ（P＞0.05）.The expressions of ColⅠand Col Ⅲ protein were ranged between 0-10%.There was no significant difference among different groups either（P＞0.05）. Conclusion Chronic fluorosis has no obvious effect on myocardial collagen metabolism of rats and myocardial collagen is not likely the main target of fluoride.

Objective To develop a sensitive,specific,rapid and easy method for detecting E.coli O157∶H7 and other Shiga toxin-producing E.coli in food and clinical specimens. Methods A circular probe and capture probe specific for Shiga toxin-2 (stx2) gene had been synthesized and was used for determining the sensitivity of ramification amplification method(RAM). Different serotypes which contained stx2 gene,including an E.coli O157∶H7, an E.coli O46∶H38, an E.coli O111∶NM,an E.coli O22∶H8 and E.coli ATCC23716(stx2 gene negative) were used for determining the specificity. All strains were detected by RAM to determine whether they contained stx2 gene.Real-time RAM was further studied to detect stx2 gene. Results The lowest number targets detected by RAM assay was 10 copies of stx2, indicating that RAM assay was as sensitive as conventional PCR. The result of specificity showed that different serotypes of strains were all positive for stx2 gene,while nonpathogenic E.coli ATCC23716 was negative.Amo ng 32 isolates, 28 STEC isolates containing stx2 gene were positive by RAM assa y, while others were negative.The RAM results were 100% in concordance with that of PCR.The real-time RAM results also showed that as many as 10 bacteria can be detected and the time of appearance of detectable signal was depended on the target concentration.Conclusion RAM assay can offer an alternative method for PCR to detect E.coli O157∶H7 and STEC in all types of specimens because of its simplicity and isothermal amplification.

Objective To investigate the possibility of constructing artificial epiphyseal plate using combined scaffold. Methods The jelly-like cartilaginous tissue was constructed by mixing epiphyseal plate chondrocyte, 0.3% collagen Ⅰ, 1 000 U·mL^-1 thrombin and 20 g·L^-1 cryodesiccant human fibrinogen. Paraffin sect ion, HE stain, toluidine blue stain and immunohistochemistry were performed regularly. Results Chondrocyte culture used collagen-fibrous protein combined scaffold was baby pink jelly-like, and the cells distributed uniformly. After 3 days, suspended matter took on milkiness, transmittancy depressed. It contracted to 7 mm while hardness increasing. By the method of HE staining, the chondrocyte was round with hyperchromatism of the nuclear, and cartilage matrix was stained powder blue within cartilage cavities in which homologue cell presen ted. The number of cells decreased gradually from surface layer in which the cel l density was biggest to deep zone. The cells were arranged by strata sequence, but didn′t lined as those of epiphyseal plate. There were brown yellow substanc e in cell periphery showed by immunohistochemistry result, which meaned collagen Ⅱ was positive. Conclusion The effect of constructing artificial epiphyseal plate using the combined scaffold is satisfied.

Objective To investigate the shortest exposure time resulted in temporary threshold shift and a sensitive objective evaluated method in long-term noise exposures, through exploring the effect of various durations of noise exposure on auditoiy brainstem response (ABR) and distortion product otoacoust ic emissions (DPOAE) in Rats. Methods Forty-two Wistar rats were divided into seven groups randomly; they were exposured to 115 dB SPL broad-band noise (12 h on/12 h off) for duration of 3,6,9,12,15 and 21 days and the ABR and DPOAE 24 h after cessat ion of the noise and again after a period of 6 weeks were tested. Results ABR thresholds were significantly elevated in all expos ure duration groups（P<0.005）. The results of linear regression analysis between ABR threshold and the duration of noise exposure were as follows: 4 h-click y=0.79x+83.81,R²=0.24; 24 h-TB y=1.26x+90.76,R²=0.40; 6 weeks-click y=1.26x+69.37,R²=0.32; 6　weeks-TB y=1.41x+82.33,R²=0.30；DPOA E response only showed significantly changes in early exposure duration. The mag nitude of DPOAE in 3 d group was not significantly compared with that of 21 d g roup. Conclusion The longer is the time of noise exposure, the smaller of the possibility for the hearing to regain. The shortest exposure to noise that induced TTS is within three days; ABR seems more sensitive to noise exposure duration than DPOAE.

Objective Objective To study the effect of Mycobacterium tuberculosis (Mtb) on IFN-γ signaling pathway in macrophages. Methods Humans and mice macrophages were isolated and infected by Mtb, condition medium were collected after a few days culture; Macrophage surface expression of FcγRI (human) or MHC Ⅱ (mouse) was measured by flow cytometry to explore the effect of condition medium on macrophage response to IFN-γ. Results Surface expression of FcγRI in human macrophages cultured in condition medium was significantly decreased compared to those without condition medium (P<0.05); Similarly, surface expression of MHC Ⅱ in mouse macro phages cultured in the condition medium was significantly decreased compared to those without condition medium (P<0.05); Condition medium cultured with ant i IL-6 antibody improved macrophage response to IFN-γ; Surface expression of MHC Ⅱ in mouse macrophages cultured in condition medium with anti IL-6 antibody was significantly increased compared to those cultured in condition medium alone (P<0.05). 　 Conclusion Mtb induced condition medium can inhibit macrophage surface expression of FcγRI or MHC initiated by IFN-γ；At meanwhile, the results also show that IL-6 may play an important role in condition medium which c an inhibits macrophage response to IFN-γ, and indicate that IL-6 may have a novel function in the Mtb infection.

Objective To identify the lectrophysiological relationship between dorsal raphe(DR) and habenular nucleus(Hb). Methods Using the methods of microinjection and electrical stim ulation and extracellular discharge recording to observe the discharge of Hb and DR. Results Of responsive neurons in Hb to DR blocked, 58% in the LHb was activated and 56% in the MHb was suppressed. Of responsive neurons to DR stimulated, 52% in the MHb was activated and 61% in the LHb was suppressed. The neurons of DR were significantly excited (53%) when Hb was blocked. The neurons of DR were depressed (62%) when Hb was stimulated. Conclusion The experimental results demonstrate that there are close functional links between DR and Hb.

Objective To investigate the influences of 5-azacytidine (5-Aza) with different concentrations on mesenchymal stem cells (MSCs) proliferation and differentiation into myoblasts. Methods Bone marrow-derived MSCs of 4 weeks Wistar rats were separated and purified,and then treated with 5-Aza with different concentrations. The growth ability of cell was assayed with methyl thiazolyl tetrazolium (MTT) method. The expression of skeletal muscle actin in MSCs was determined with reverse transcription polymerase chain reaction (RT-PCR) and electrophoresis after MSCs were induced.Results The cell proliferation was not affected by 0 and 1 μmo l·L^-1 5-Aza . There were expressions of skeletal muscle actin treated w ith 3-12 μmol·L^-1 5-Aza. Some cells were obviously enlarged at the 9th day after induction and myotubu-like cells were found at the 12th day when treated with 9 and 12 μmol·L^-1 5-Aza. 20-30 μmol·L^-1 5-Aza induced the toxic effect on proliferation of MSCs. With the increase of concentration, the proliferation ability of MSCs was weakened.Conclusion 5-Aza affects the expression of regulatory gene to the stem cells and regulate MSCs to orientationally differentiate into myotube- like cells.

Objective To investigate the expression of peroxisome proliferator activated receptor γ（PPARγ）in atherosclerosis. Methods 16 rabbits were randomly divided into cholesterol diet group and normal diet group（control group）（n=8）.Atherosclerosis models w ere built in cholesterol diet group. Serum TC, TG, LDL in rabbits were measured after bred for 0, 8 and 16 weeks in two groups. After 16 weeks the aortas were harvested for histomorphometry observation and PPARγ immunohistochemistry analysis. Results The differences of TC,TG and LDL levels between two groups were not significant in 0 week （P>0.05）; and after 8 and 16 weeks, co mpared with 0 week and control group, serum TC,TG,LDL levels in cholesterol diet group were significantly increased（P<0.01）.The aortas intima plaques were formed in cholesterol diet group by the aorta histomorphometry observation，plaque/intima size in cholesterol diet group and control group（P/I）were(28.12±3.77)% and 0,respectively（P<0.01）,intima thickness were (67.47±7.13)μm and (4.45±0.58)μm（P<0.01）,intima/medium thickness (I/M) wer e 0.878±0.370 and 0.054±0.007（P<0.01）.With SP method and DAB dye，PPARγ expressions in cholesterol diet group and control group were (14.38±2.58)% and (7.82±0.96)%, respectively（P<0.01）. Conclusion PPARγ can express excessively in atherosclerosis location, it may involve in the process of atherosclerosis.

Objective To study the effects of heroin on adenosine deaminase (ADA) in small inteseine and plasma uric acid content in rats. Methods The rat models injected with heroin and forbidden to heroin were set up. The blood samples from the abdominal aorta were obta ined，then the plasma uric acid content was detected. 10% homogenate of small intestine wa s prepared after the rats were killed, then the ADA activity was determined. Results Compared with the control group, the content of plasma uric acid of the rats increased significantly after administration of heroin for 9 d (P<0.01)，the ADA activity in small intestine increased (P<0.05). Compared with 9　d heroin administration group，the content of plasma uric acid in 8　d heroin withdrawal group decreased,but the difference was not significant (P>0.05); and the ADA activity in small intestine decreased significa ntly （P<0.05）.Conclusion The results may suggest that long-term heroin administration can enhance the catabolism of purine nucleotide in small insestine by increasing the activity of ADA.

Objective To record the electroenciphalogram(EEG) in hippocampus or parietal cortex and then compare them in epileptic rats kindled by kainic acid(KA) microinjection into nucleus amygdalae. Methods KA was microinjected(1 μg) into right nucleus amygdalae of male Wistar rats with stereotaxic technique to establish kindling epileptic model. Observation of praxiology was carried while recordin g of the EEG in hippocampus and parietal cortex were performed successively for 6 h after KA injection. Results Many kinds of epileptic discharges presented in the kin dled rats, especially the multiple spike waves were common. There were no significant differences in the latency to onset, period of discharge(PD) per hour, total PD in 6 h and the amplitude of wave between EEG of hippocampus and that of parietal cortex. Conclusion The changes of EEG in rats kindled by KA microinjection into nucleus amygdalae are consistent with that of temporal lobe epilepsy. Seizures are quickly spread inter-cortex. The epileptic discharges of hippoc ampus opposite to kindling focus are highly coincident to that of parietal cort ex. Motoring the EEG of parietal cortex may refer to observe the changes in electrophysiology for this model.

Objective To study the influences of nitric oxide (NO) and superoxide dismutase (SOD)in genesis and progress of delayed neuronal death (DND) in rats. Methods Wistar rats were used to establish the models of four-vessel occlusion (4VO) and were treated with small dose of nitric oxide synthase (NOS) inhibitor ［N-nitro-L-arginine (NNLA)］. The levels of NO and SOD in the brain tissues of the models were determined by spectrophotometry. Results The level of NOx (NO₂+NO₃)in the experiment group was not significantly increased compared with control group (P＞0.05) . When reperfusion time was prolonged, the level of NO in the experiment group was decreased gradually and was lower than that in control group (P＜0.05 or 0.01）. The level of SOD in hippocampus region was decreased distinctly(P＜0.05 or 0.01）and was not influenced by NNLA. The relativity analysis showed that the levels of NO and SOD in brain tissues were highly relative (r=0.9624,P＜0.05 or 0.01）. Conclusion Small dose of NOS inhibitor has the effect to protect neuron in DND rats. SOD can not only clean out superoxide negativeion,but also eliminate reactive oxygen species of NOx.

Objective To investigate the correlations between vascular dementia and neurotrophin-3(NT-3)，fibroblast growth factor(FGF). Methods Female and male Wistar senile rats were divided into the control group and the operation groups at random. The operation groups consisted of the cerebral ischemia 15 min group and ischemia 15 min reperfusion groups in which the rats were reperfused at 0，1st，6th hour and on 2nd，4th，9th day with five rats per group. The expressions of NT-3 and FGF in hippocampus and thalamus were examined in different periods of ischemia-reperfusion with ABC method of immunohistochemistry. Results There was a little expressions of NT-3 and FGF in hippocampus and thalamus of normal senile rats;the expression of NT-3 in hippocampus increased quickly and steadily from 1 h to 4 d after ischemia-reperfusion; there were signifi cant differences compared with control group （P<0.05）. The expression of NT-3 in hippocampus reduced on 9th day after ischemia-reperfusion.There were no differences compared with control group （P>0.05）. The expression of FGF in hippocampus increased from 6 h to 2 d after ischemia-reperfusion（P<0. 05）, the expression of FGF increased further on 4th day （P<0.01）. Then it reduced sharply, there were no differences compared with control group （P>0.05）. FGF in thalamus began to increase on 2nd day after ischemia-reperfusion and there were significant differences compared with control group （P <0.05）. Then the expressions of NT-3 and FGF in thalamus decreased, there were no differences compared with control group （P>0.05）. Conclusion Hippocampus has the protective mechanism of NT-3 an d FGF against ischemia injury which is produced quickly and has endurable effects, but thalamus lacks the good protective mechanism of NT-3 and FGF.

Objective To study the inhibitory effect of 20(S)-ginsenoside Rg3 on tumor growth and its mechanism. Methods MTT assay and the model of B16 melanoma were used to observe the effect of 20(S)-ginsenoside Rg3 on B16 melanoma growth and its mechanism was analysed by assay of tumor-induced angiogenesis and flow cytometry. Results In vivo, the average weight of B16 melanoma in each Rg3 group was lower than that in control group (P<0.01) and the number of vesssels oriented toward the tumor mass was less than that in untreated control (P<0.01). In vitro, the results of MTT assay displayed that ginsenoside Rg3 inhibited the growth of B16 melanoma cells and the IC₅₀ value was （7.76±0.46）mg·L^-1. Rg3 blocked B16 melanoma cells in G₀/G₁ and S stages. Conclusion 20(S)-ginsenoside Rg3 can obviously inhibit the tumor growth and the effect may be related to its inhibiting angiogenesis and blocking cell mitosis.

Objective To study the ameliorative effects of pseudoginsenoside GQ (PGQ) on electrocardiogram changes in rats with acute myocardial ischemia induced by isoproterenol(ISO).Methods 50 Wistar rats were divided into five groups (n=10) randomly. Group Ⅰ: i.v. 0.9% sodium chloride water liquor; Group Ⅱ: i.v. verapamil hydrochloride of 0.2 mg·kg^-1; Group Ⅲ: i.v. 3.0 mg·kg -1 of PGQ; Group Ⅳ: i.v. 6.0 mg·kg^-1 of PGQ; GroupⅤ: i.v. 12.0 mg·kg^-1 of PGQ. Myocardial ischemia was induced by intravenous injection of ISO (2.0 mg·kg^-1). ST-segment changes in the lead Ⅱ,precordial in lead Ⅴ₁ and precordial in lead Ⅴ₂ were examine d. Results In lead Ⅱ, lead precordial Ⅴ₁ and lead precordial Ⅴ₂， ST-segment elevation of three PGQ groups were not obvious at every t ime point, the extents of ST-segment elevation were lower than those in sodium chloride water liquor group（P<0.05，P<0.01，P<0.001）. Conclusion PGQ has the ameliorative effects on electrocardiogram changes in rats with ISO-induced acute myocardial ischemia.

Objective Through observation of liver ultrastrutures in obstructive jaundice to study its injury mechanisms. Methods Seventy-two Wistar rats were randomly divided into 3 groups:experiment, sham and normal groups (n=24). Wistar rat models with obstructive jaundice were made in experiment group. The liver ultrastructure in three groups were observed by the transmission electron microscope. The changes of the liver function were analyzed with 7150 autobiochemistry analytic instrument. Results The nucleus morphology of the hepatocytes was abnormal ,the nucleus was pyknotic, the mitochondria crista was broken. The endoplasmic reticulum was reduced and swelled. Bile stasis was present within dilatation canaliculi.The results of biochemistry measurement of liver function indicated that the differenes of TB IL, TRA, ALT, and AST were statistically significant between experiment and sh am, normal groups (P<0.01). Conclusion Endotoxemia, bile acidemia, hepoemdia of the liver and Kupffer cells function injury may relate with the liver injury of obstru ctive jaundice.

Objective To study the effect of anti-mutation of Chinese medicinal herb Polygala tenuifolia willd on T lymphocyte proliferation in spleen. Methods The micronucleus test of mouse bone marrow cell (MNT):thirty mice were divided into six groups (n=5)， negative control（NS）,cyclophosphamide group（CP 3.0 mg·kg^-1），Polygala tenuifolia willd antimutagenesis groups（Polygala tenuifolia willd with dosage of 0.5,1.0,2.0,4.0 g·kg^-1+ CP 30 mg·kg^-1）. The improved method was used to detect the micro-nuclei frequency. Lymphocyte transformation test: twenty-four mice were divided into four groups (n=6)， saline control ,CP 　control（3.0 mg·kg^-1）,Polygala tenuifolia willd （2.0 g·kg^-1），Polygala tenuifolia willd+CP（2.0 g·kg^-1 Polygala tenuifolia willd+CP 30 mg·kg^-1） group. MTT assay was used to calculate the stimulation index(SI). Results The micro-nuclei frequency was significant difference between Polygala tenuifolia willd antimutagenesis groups and CP groups (P<0.05)；the SI in saline control,Polygala tenuifolia willd,Polygala tenuifolia willd+CP,CP groups were 2.11±0.13,2.37±0.18,1.97±0.06 and 1.34±0.19,resp ectively, the difference of SI was significant between Polygala tenuifolia willd and saline control groups（P<0.05）. The difference of SI was significant between Polygala tenuifolia willd+CP and CP groups（P<0.05）. Conclusion Polygala tenuifolia willd may protect genetic material from injury caused by CP, namely,showing effective antimutagenesis and ye t Polygala tenuifolia.willd may increase the reactivity of mouse spleen cells to ConA and could improve reactivity reduction induced by CP, namely Polygala tenuifolia willd could enhance lymphocyte function.

Objective To observe the effect of acrous g ramineus and its main component α-asarone on eclamptic behaviors and frontal lobe discharge in the young rats with experimental epilepsy induced by pentylenetetrazol (PTZ).Methods Young Wistar rats were divided into five groups randomly including normal control (N), epileptic control (A), phenobarbital sodium (B) , acrous gramineus (C), α-asarone （D）. Except normal control, other animals were ip. PTZ 60 mg·kg^-1 to found epileptic animal models. Rats in both normal control and epileptic control were ip. saline 0.5 mL, and other groups were ip. phenobarbital sodium 18 mg·kg^-1, acrous gramineus 2.35 g·kg^-1, α-asarone 29 mg·kg^-1, respectively. Expect normal control, other rats were ip. PTZ 60 mg·kg^{-1 again after they were treated for 7 d. Using brain solid positioner, the changes of unilateral frontal cortex electroencephalogram (EEG) in young rats were recorded as well as their ecliptic behaviors were observed. Results ①Showed as animals′ behaviors, in treated groups, the latent period of paroxysm was delayed significantly (P<0.05), the number of titanic-cloni c seizures was decreased obviously during 2 h (P<0.05), the mean duration of seizures was reduced compared with epileptic control (P<0.05). The functions of acrous gramineus and α-asarone mentioned above were weaker than phenobarbital sodium (P<0.05). ②Showed as EEG, in treated groups, the latent of epileptic brain wave in frontal cortex was elongated (P<0.05), both the frequency of abnormal discharge in 2 h and the amplitude of brain wave were descended compared with epileptic control (P<0.05). The influences of acrous gramineus and α-asarone on EGG were more significant than phenobarbital sodium (P< 0.05). Conclusion Both acrous gramineus and α-asarone can suppress seizure and abnormal discharge of frontal cortex induced by pentylenetetrazol in young rats availably.}

Objective To observe the protective effects of Diemailing Injecti on (DMLI) on myocardial ischemia-reperfusion injury in rats. Methods The myocardial ischemia-reperfusion model was induced by 30 min left a nterior descending coronary occulusion and 24 h reperfusion in open-chest anesthetized rats. The changes of aspartate aminotransferase(AST), lactate dehydrogenase(LDH), MB isoenzyme of creatine kinase (CK-MB), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities and malondialdehyde (MDA) contents in serum, and prostacycline(PGI₂) and thromboxane A₂ (TXA₂) levels in plasma were determined. Results In rats treated by DMLI (with dose of 2.5,5 and 10 mL·kg^-1 i.v. at 30 min after coronary occulusion),the myocardial ischemia size (MIS) was significantly reduced, the AST，LDH and CK-MB activities in serum and the TXA₂ leve l in plasma were declined, while PGI₂ level in plasma and PGI_{2/TXA2 ratio were increased significantly. In addition, the LPO content in serum declined, SOD and GSH-Px activities in serum were increased markedly. Conclusion DMLI has protective effects on myocardial ischemia-reperfusion injury through improving free radicals metabolism, decreasing TXA2 level in plasma, increasing PGI2 level in plasma and PGI2/TXA2 ratio.}

Objective To observe the preventive effect of Polygala tenyifolia willd on damage of genetic materials in male mouse reproductive cells. Methods Unscheduled DNA synthesis（UDS）test in male mouse reperoductive cells was used. Results There was no significant difference (P>0.05) while UDS was induced by Polygala tenyifolia willd with various doses comparing w ith normal control. Nevertheless, Polygala tenyifolia willd in doses ranged from 1.0 to 4.0 g·kg^-1 inhibited strikingly UDS induced by Pb(CH₃COO)₂ (P<0.01),and the inhibitory effect was enhanced with the incresse of the concentration. However the effect did not change when the dose reached a certain concentration. Conclusion Polygala tenyifolia willd shows a protective effect, namely antimutagenesis，on injury of genetic materials in reproductive cells of male mouse.

Objective To improve the efficacy of the Nd∶YAG laser-induced chorioretinal venous anastomosis in the rabbits′eyes. Methods Immediately after branch retinal vein occlusion(BRVO) model was made by argon laser in 40 eyes of 20 normal rabbits， the rabbits were divided into two groups. In the first group （experimental group），there were 10 rabbits treated with the argon laser followed by the Nd∶YAG laser induced chorioretinal venous anastomosis. In the second group（control group）,the rabbits were treated only with the argon laser induced chorioretinal venous anastomosis. Results An anastomosis in experimental group was formed in 6 rabbits within 4-6 weeks after treatment while no anastomosis was formed in control group. During 2-6 months the hemorrhage of retina or vitrous with laser photocoagulation were absorbed within 8 weeks and no other severe complications were found. Conclusion Applying Nd∶YAG laser can improve the efficacy of chorioretinal venous anastomosis in the rabbits′eyes.

Objective To observe the changing rules of skeletal muscle and motor end plate of the denervated rats. Methods Fifty-four adult Wistar rats were used and separated randomly in nine groups. After established the model of denervated gastrocnemius muscle by cutting off the sciatic nerve， the muscle wet weight， diameter，cross section area of myocyte and motor end plate were measured. Results 2 weeks after denervated， the wet weight reduced to 52.1% of the normal， until 16 weeks later， it was maintained at about 25% of the normal until to 24 weeks. The diameter and the cross section decreased progres sively with prolongation of time. 2 weeks after nerve resected， the diameter wa s 64.87% of the normal， and it was only occupied 25.06% of the normal 20 week s later. The cross section changed as the changes of the diameter. The motor en d plate was normal in the first four weeks. After that， the motor end plate losed the normal shape and formed neurofilament. 16 weeks after operation， the neurofilament began to decrease. Conclusion The skeletal muscle atrophies and the motor end plate cataplasia take place after the nerve injury, the changes are getting serious with prolongation of time. The nerve repair operation should be made as early as possible.

Objective To study the simple preparation method and structure of nano-hydroxyapatite/collagen composite, to investigate new substitute of repairing bone for using in tissue engineering. Methods Porous nano-hydroxyapatite was made of Ca(OH)₂ and H₃PO₄. Collagen was drawn from fresh adult bovine tendon. The two materials were prepared into biomembrane through the glutaraldehyde and f reeze-drying. The crystallite phase, micro-morphology, structure, crystallite size of the composite were examined by XRD and scanning electronic microscop（SEM）. Results The results showed that the composite structure was porous and consisted of nano-hydroxyapatite （10 nm×50 nm-20 nm×80 nm）and collagen fiber. The crystallite phases and size of the composite was similar to that of natural bone. Conclusion The porous nano-hydroxyapatite /collagen composite is expected to be an ideal substitute of repairing bone.

Objective To observe heterotopic osteogenesis of autogenous marrow stromal cells on β-tricalcium phosphate（β-TCP） scaffold,and the effects of recombinant human bone morphogenetic protein-2(rhBMP-2) and transforming growth factor-beta (TGF-β) on osteogenesis. Methods The Newzealand rabbits autogenous marrow stromal cells were cultured by mineralized condition medium. The cell suspension was collected. All the cells with concentration of 5×106·mL^-1 we re divided into four groups. Group A: rhBMP-2（15 μg）/TGF-β（30 ng）were added; group B:TGF-β 30 ng was administered; group C:without growth factor as control; group D:continual cultured and no loading on β-TCP. MSCs in group A,B,C were planted on β-TCP.Alkaline phosphatase (ALP) activity was examined by histochemistry. Then MSCs loading on β-TCP were embedded in autogenous subcutane ous. Four weeks later, the osteogenesis was observed by HE staining, typeⅠcollagen and bone morphogenetic protein synthesis were examined by immunohistochemistry stains. Results The ALP activities in groups A and B were significantly higher than those in group C and D （P<0.01）. Five weeks later, some strip new bone formations were observed by HE staining in all groups, the bone matrix area in group A was significantly bigger than that in group B（P<0.01）. The average gray degree of type Ⅰ collagen in groups A and B were lower than that in group C. There was not obviously difference of bone morphogenetic protein expression between group A, B and C. Conclusion β-TCP and MSCs can promote heterotopic osteogenesis, rhBMP-2 and TGF-β have the stronger effect to enhance osteogenesis than rhBMP-2.

Objective To detect the association between metallothionein (MT) 1X gene polymorphism and type 2 diabetes mellitus (T2DM) in Chinese Han people. Methods The techniques of PCR and restriction fragment length polymorphism (RFLP) were used to examine MT4 gene (rs4531729) C→T mutation in 132 patients with T2DM and 137 unrelated healthy individuals, and then the mutation frequency was statistically computed.Results The alleic frequency (C) of MT 1X gene mutation was fou nd no difference between the diabetics and non-diabetics (χ²=1.508,P>0.05),and the distribution rates of CC, CT and TT also had no significant difference (χ²=1.581,P>0.05) between the case and the control.Conclusion There′s no association between the alleic gene of M T 1X gene mutation and T2DM, and the mutation genotype may not increase the risk of suffering from T2DM.

Objective To investigate the association between KCNJ10 gene polymorphism and schizophrenia.Methods PCR based on restriction fragment length polymo rphism (PCR-RFLP) assay was used to detect the genotype of rs1186679 locus on K CNJ10 among 90 Chinese Han family trios consisting of fathers, mothers and affec ted offsprings with schizophrenia. Results The goodness-of-fit chi-square test showed that the genotypic distribution of rs1186679 didn′t deviate from Hardy-Weinberg equilibrium. Haplotype relative risk (HRR) test did not show a genetic association bet ween rs1186679　and schizophrenia (χ²=0.775, P>0.05). Transmission disequilibrium test (TDT) did not show a significant biased transmission from pa rents to affected offsprings (χ² =1.025, P>0.05). Conclusion The rs1186679 locused on KCNJ10 has no association w ith schizophrenia in Chinese Han people.

Objective To investigate the distribution of active MMP-2 in human breast cancer tissues and its relation with invasion and metastasis of breast cancer. Methods Gelatinolytic activity was detected with tissue zymogra phy and in situ zymography technique using DQ-gelatin.Results Tissue zymography results showed only MMP-2 exited the mode of activation, gelatinolytic activity was showed in the breast cancer cell s. In situ zymography using DQ-gelatin revealed that strong green fluoresce nce was present in the carcinoma cell nests. Meanwhile, green fluorescence was present in peri-carcinoma, but the intensity of fluorescence was not the same. There were statistically significant correlations between the intensity of fluorescence and tumor grad e, metastasis. Conclusion Gelatinolytic activity is produced by MMP-2 in brea st cancer. The active form of MMP-2 localizes in the cancer cells and plays a vital role in invasion and metastasis in breast cancer.

Objective To study the expressions of matrix metalloproteinases-2 (MMP-2) and its tissue inhibitor-2 (TIMP-2) and microvessel density (MVD) in human endometrial carcinoma tissues and their relations with the invasion a nd metastasis of endometrial carcinoma. Methods Immunohistochemistry was used to measure the MMP-2, TIMP-2, and MVD protein levels in 33 patients with endometrial carcinoma and 8 normal endometrial samples. Results The MMP-2, TIMP-2 and MVD protein expressed in both normal endometrial samples and endometrial carcinoma. The positive expression proportions of MMP-2, TIMP-2, and MVD protein in endometrial carcinoma tissues we re higher than those in normal endometrial samples (P<0.05),but they did not show different expression with the histologic grade (G₁,G₂ and G₃). Semi-quantitative analysis revealed MMP-2 staining scores in tumor cells were significantly associated with the presence of surgical pathologic staging (P<0.05),while TIMP-2 staining scores in tumor cells were significantly associated with the depth of myometrial invasion (P<0.05). MVD staining scores in tumor cells were also significantly associated with the depth of m yometrial invasion (P<0.05) . But these factors had no significant association with the presence of lymph node metastasis. Conclusion The expression maladjustment between MMP-2 and TIMP-2 may accelerate the tumor invasion and metastasis; the increasing expression of MVD in tumor cells may be associated with the depth of myometrial invasion, which suggests that angiogenesis plays an important role in invasion and metasta sis of endometrial carcinoma.

Objective To investigate the expressions of Stat3, CyclinD₁ and proliferating cell nuclear antigen (PCNA) in non-small cell lung cancer (NSCLC),and analyze their relationships with carcinogenesis and progression of NSCLC. Methods SP immunohistochemistry was used to detect the expressions of Stat3, CyclinD₁ and PCNA in 45 cases of NSCLC. Results The positive rate of Stat3 was 71.1 % in NSCLC. The expression intensities of Stat3 was not associated with different tumor sizes and histological type, but it was associated with the expression intensities of CyclinD₁ and PCNA （r＝0.531, P<0.01；r＝0.618, P<0.01）and it was re lated to histological grade and lymph node metastasis（P<0.05）.Conclusion The expression of Stat3 is abnormal in NSCLC. Stat3 may involve in regulation of CyclinD₁ in NSCLC and accelerate proliferation of NSCLC, it may play an important role in generation of NSCLC.

Objective To study and analyze the significance of the changes of plasma level of neuropeptide (NPY) in the patients with chronic hepatic disease. Methods The plasma levels of NPY were determined by radioimmuno assay (RIA) in 17 chronic hepatitis patients, 48 hepatic cirrhosis patients and 21 healthy controls. 48 patients with hepatic cirrhosis were divided into three groups: hepatic cirrhosis without ascites(n=25), hepatic cirrhosis with ascites but without hepatorenal syndrome (HRS) (n=19), hepatic cirrhosis with HRS(n=4). 48 hepatic cirrhosis patients were also divided into another three groups according to liver function:Child A (n=20), Child B(n=16), Child C(n=12). Results Compared with healthy controls ,the plasma levels of NPY in chronic hepatitis patients and hepatic cirrhosis patients without ascites had not significant statistical difference (P>0.05); but plasma levels of NPY in patients with hepatic cirrhosis and ascites or those with HRS had significant difference (P<0.05). No corralation was found between plasma level of NPY and liver function, but the plasma level of NPY was correlated directly with plasma levels of BUN and Cr（r=0.872, P<0.01; r=0.857, P<0.01）.Conclusion The determination of plasma level of NPY in patients with chronic hepatic disease may be one of important indexes in evaluation of disease and prognosis.

Objective To investigate the correlations of serum matrix metalloproteinase-2,-9 (MMP-2,MMP-9) contents and tissue expressions in hepatocel lular carcinoma with tumor invasion and metastasis. Methods Serum MMP-2,MMP-9 contents were detected in 40 patie nt with hepatocellular carcinoma and 20 healthy controls by ELISA; the expressi ons and distributions of MMP-2 and MMP-9 in 40 patients and 10 normal tissues were detected by immunohistochemical method. Results Serum MMP-2,MMP-9 contents were significantly elevated in cancer samples compared with normal serum (P<0.01), the significant difference was found between contents in the presence and the absence of lymph node metastasis (P<0.05).In hepatocellular carcinoma, the expressions of MMP-2,MMP-9 were increased significantly compared with normal tissue.The expressions of MMP-2,MMP-9 were correlated with histological grade and lymph node metast asis (P<0.05). Conclusion The serum of MMP-2 and MMP-9 contents and their expressions may pr ovide reliable information for hepatocellular carcinoma prognosis.

Objective To select materials with high quality and safety and evaluate the clinical value of multi-point forming technique in skull repairing. Methods 161 patients suffered from skull defect had been cured in our hospital, within them 43 patients were treated with multi-point forming technique in titanium mesh shaping, 19 patients with traditional handwork shaping, 99 patients with bone-like concrete (acrylate). The following aspects were analyzed and compared：neurosurgeons′ shaping workload before operations,operative time, approving scale on shaping and complications after operation. Results Repairing skull by titanium mesh with the technique of multi-point shaping significantly shortened the average operative time to （30±6） min, compared with acrylate group (70±18 min) and titanium mesh trad itional handwork shaping group (50±11 min) (P<0.01); and decreased the neurosurgeons′　working intensity of shaping time to （5±1 min）, compared with acrylate group (30±5 min) and traditional handwork shaping group (20±3 min) (P<0.05); and improved the patients′ approving scale to 100%, compared with acrylate group (91%) and traditional handwork shaping group (95%) (P<0.05).Conclusion The quality of titanium mesh is obviously better tha n acrylate, the effect of repairing skull with multi-point forming technique is superior to handwork shaping.

Objective To investigate the relationship between WBC concentration in semen and sperm morphological parameters. Methods Semen samples were collected from 278 infertile men. WBC in semen was detected through WBC count by microscope.Sperm morphology was evaluated using artificially modified method by Automa ted Sperm Morphology Analyzer.Acrosomal morphology was analysed with modified P apanicolaou staining.Results The percentage of morphologically normal sperm and t he rate of acrosomal intactness in leukocytospermic group were both significantl y lower than those in nonleukocytospermic group（P<0.05）.The WBC concentration in semen had significantly negative correlation with the percentage of morphologically normal sperm and the rate of acrosomal intactness（r=-0.243, P<0.01; r=-0.248, P<0.01）. Conclusion Excessively high WBC concentration in semen could affect morphology of sperm and acrosomal intactness.

Objective To study the action of Mycoplasma genitulium (Mg) in pathogenesis of chronic non-bacterial prostatitis. Methods Mg culture and polymerase chain reaction (PCR) were per fomed on prostatic secretion specimens from 987 patients with chronic non-bacte rial pristatitis and 125 healthy men. Results ①The positive rate of Mg in the prostatic secretion from the 987 patients was 31.5% (302/987). Among the 302 positive cases, 153 cases had other pathogens, of which Ureaplasma urealyticum (Uu) was the most (128).②In the controls, the positive rate of Mg was 2.53% (3/125). The difference between the two groups was statistically significant (χ²=44.32,P<0.01). Conclusion The infection rate of Mg in patients with chronic non-bacterial prostatitis in native regions is higher and Mg may be one of the pa thogenic factors of chronic non-bacterial prostatitis.

Objective To study the relationship between reactive oxygen species（ROS） in polymorphonuclear granulocytes （PMN），SOD of peripheral blood and tumor′s genesis and aggression.Methods Sixty patients with cancer and sixty healthy persons were chosen. FACS and pyrogallol auto-oxidation test were used to detect ROS in PMN, SOD activity of peripheral blood. Results ROS in PMN in cancer group (387.12±2.87 MFI) was higher than that in control group　(112.34±3.64 MFI) (P<0.01)；ROS in PMN in primary cancer group (493.56±3.44 MFI) was higher than that in metastatic cancer group (269.42±4.10 MFI) (P<0.05); SOD level in peripheral blood in cancer group (218.80±4.59 U·g^-1 Hb) was lower than that in control group (320.57±3.35 U·g^-1 Hb)(P<0.05), and SOD level in peripheral blood in primary cancer group　(189.41±2.85 U·g^-1 Hb) was lower than that in metastatic cancer group (256.13±3.66 U·g^-1 Hb) (P<0.05). Conclusion There is metabolic disturbance of ROS in patients wi th cancer and ROS may be associated with the stage of cancer.

Objective To construct the expression vector and express the recombinant human L-selectin in prokaryotic system, and purify the aimed protein for the preparation of rabbit anti-human L-selectin polyclonal antibody. Methods The partial cDNA of human L-selectin was amplified from the open reading frame (ORF) of N-terminus sequence of L-selectin contai ning 153 amino acids by PCR, then cloned into the prokaryotic expression vector pQE40 at restriction sites BamH Ⅰ and Hind Ⅲ. The recombinant expression plasmid pQE40-L-selectin was transformed into E.coli M15 for expression. The fusion protein including 6-His-tag was purified by Ni-NTA chromatographic column and analysed by SDS-PAGE. The purified protein was used to immune rabbit for pre paring polyclonal antibody. Western blotting analysis and dot immunoblot assay (DIBA) were used to test the titer of the antiserum. Results The expression plasmid pQE40-L-selectin was constructed and confirmed with restriction enzyme digestion. The quantity of the purified protein from E .coli M15 was 600 mg·L^-1. By immuning the rabbit, the polyclonal antibody was successfully prepared. The results of dot immunoblot assay (DIBA) showed that the antiserum had the high titer (1∶1 000). Conclusion The recombinant human L-selectin protein can express with high efficiency in E.coli M15. The prepared polyclonal antibody ha s a high titer.

Objective To isolate and identify the phenotype and biological characteristics of fetal liver derived mesenchymal stem cells(MSCs) as well as the MSCs derived from fetal bone marrow. Methods Mononuclear cells from fetal liver were collected by Percoll gradient centrifugation. The low density cells were cultivated and expanded in MSCs media. Phenotype was analyzed by FACS flow cytometer and characteristics of the multipotent were observed. Results The expressions of CD29,CD44,CD73（SH-3，4）,CD105（SH -2）,CD166,HLA-ABC in MSCs from fetal liver and fetal bone marrow were positive，CD35,CD45,HLA-DR were negative. MSCs from fetal liver and bone marrow were successfully cultivated and differentiated into osteoblast and adipose in vitro. As the quantity of MSCs increased,their inhibitory effects on transformation of lymphocytes in peripheral blood enhanced. Conclusion MSCs existing in fetal liver and bone marrow can be isolated successfully.

Objective To establish a fast and accurate method to examine the Listeria monocytogenes in foods. Methods The polymerase chain reaction (PCR) method was adopted to amplify hlyA gene of Listeria monocytogenes. Eighty food samples of Changchun city were detected by PCR method. Results The target fragment of hlyA gene appeared at 706 bp. 12 positive results was gained in all detected samples,the positive rate was 15%. This result was identical with the detection result obtained by CDC of Jilin Province with international standard method,the coincidence rate was 100%. Conclusion PCR method is more concise, rapid, sensitive than other methods in detecting Listeria monocytogenes in food, the specificity is the highest. This method is applicable for basic sanitation supervision department to investigate and supervise the situation of Listeria monocytogenes contamina tion in food.

Objective To investigate the incidence and related risk factors of Eales disease in healthy young men. Methods 3 288 recruits from Beijing region were taken part in t he investigation. Based on the diagnostic standard forwarded by Jyotirmay Biswas and modificated slightly by ourselves, all the partners were examined by binocular indirect ophthalmoscope. The diagnosis was made by the appearance of the fund, and some ancillary text was used to exclude another disease that mimicking Eales dise ase. The incidences of Eales disease and the relationship between the incidences of the Eales disease and some factors such as region, diet, income, smoking, drinking and the result of PPD were analyzed by SPSS 12.0 for Windows. Results 26 were diagnosed ultimately in 3 288 recruits. 23 were suffered by single eye and 3 were suffered by double eyes. The incidences of Eales disease was 0.8% with the credible range from 0.52% to 1.16%. There were no statistical correlation between the Eales disease and the region, diet, income，smoking (P>0.05), and there seemed to have some statistical correlation between the incidence of Eales disease and PPD (P<0.05). Conclusion There is high incidence of Eales disease in healthy young men, and it approachs to the level of the outpatient in Indian. PPD may play some roles in the incidence of Eales disease.

Objective To discuss the relationship between the infection of Toxoplasma gondii and female sterility. Methods Toxoplasma gondii serum antibody were determined in 882 women with sterility (experimental group) and 107 normal bearing women (control group) by using ELISA. At the same time the differences of the infection with Toxoplasma gondii between the ages of the sterility women were analyzed. Results The positive rate in experimental group was 15.87% (140/882), the positive rate in control group was 5.61% (6/107), remarkable difference was found between two groups(P<0.01). The infection rate in the diffe rent age groups (20-24, 25-29,30-34,35-39 and ≥40) is 5.63%,15.24%,17.91%,19.44% and 15.38%. Conclusion Toxoplasma gondii infection may be one of the factors which can cause sterility, and the infection rates at different ages have no instinct differences.

Objective To investigate and analysis the situation of malocclusion of 49 skulls of ancient resident in Xinjiang, and discuss the history evolution and the pathologic mechanism of malocclusion. Methods The investigation was carried out based on the standard of individual normal occlusion. Results Among the group of 49 skulls, the morbility rate of malocclusion was 30.61%; in Angle′s classification of malocclution,class I was th e most (73.33%); in Mao′s classification of malocclution, class I was also the most (33.33%). Conclusion The morbility rate of malocclusion of resident in bronze age is lower than that of modern resident and nearer ages resident. There is a close releationship between the human food construction and the pathogenesis of malocclution.