Objective To study the protective effects of Gross Saponins of Tribulus Terrestris (GSTT) on acute myocardial ischemia in rats. Methods An acute myocardial ischemic model was set up by ligating left coronary artery, the effects of GSTT on the serum LDH, SOD, and MDA contents in rats were observed. Meanwhile, HE staining and electron microscope observation were used to study the morphological changes of cardiomyocytes. The TUNEL semi-quantitative technique was performed to observe the effects of GSTT on apoptosis rate. Results Compared with control group, GSTT decreased the serum LDH and MDA contents（P<0.01）,while it increased the serum SOD level (P<0.01) in rats. It decreased myocardial apoptosis rate as well(P<0.01). The obvious pathological changes of cardiomyocyte structure were found in model group. Conclusion GSTT has protective effects on acute myocardial ischemia induced by ligating left coronary artery in rats in vivo, which is closely related to enhancing removement of free radicals by increasing SOD content, decreasing the leakage of LDH, and inhibiting the apoptosis of cardiomyocytes.

Objective To study the effects of Gross Saponin of Tribulus terrestris (GSTT) on neonatal rat cardiomyocyte apoptosis induced by NaCN and its mechanisms. Methods The intracellular ischemic model was set up with NaCN, the effects of GSTT on apoptotic rate and mitochondrial membrane potential(△ψm) were determined by flow cytometry. The intracellular calcium level was detected by confocus system, and the protein expression of Bcl-2/Bax were observed by Western blotting. Results The cardiomyocyte apoptosis rate induced by NaCN in GSTT group （3.31±0.24%）was decreased significantly than that in control group　(16.65±0.18%)(P<0.01).The mitochondrial membrane potential in GSTT group (158.29±0.40) was higher than that in model group (121.71±0.80)(P<0.01)，was lower than that in control group (167.10±0.70)(P<0.01). The level of intracellular calcium in GSTT group (323.73±68.10) was lower than that in model group (496.65±80.10)(P<0.01)，and was higher than that in control group (280.18±48.10)(P<0.01)). GSTT up-regulated Bcl-2 and down-regulated Bax protein expressions. Conclusion GSTT can inhibit the cardiomyocyte apoptosis induced by NaCN, and the inhibitory effect was closely related to regulation of the protein expression of Bcl-2/Bax, stability of mitochondrial membrane potential and reduction of intracellular calcium.

Objective To observe effects of Gross Saponin of Tribulus Terrestris (GSTT) on cardiac hemodynamics in isolated working rat heart. Methods The direct effects of GSTT on myocardiac contractility were observed by a pattern of Langendorff isolated,perfused Wistar rat heart through determining the hemodynamics parameters including aorta flow (AF), coronary flow (CF), heart rate (HR), arterial blood pressure(AP), left ventricular systolic pressure (LVSP), left ventricular end-diastolic pressure (LVEDP), and ±dp/dt_max . Results The LVSP，±dp/dt_max，AP， and AF were decreased as well as the LVEDP, CF were increased 5-20 min after the perfusion of GSTT with the doses of 100,150, and 200 mg•L^-1 compared with normal control, the differences were significant（P<0.05 or P<0.01）. GSTT could decreased cardiac contractility and expanded coronary in a dose-dependent manner. Meanwhile, the GSTT decreased the positive inotropic effect of higher Ca²⁺ by its negative inotropic effect and increased contractility of lower glucosi cardiac. Conclusion GSTT may cause negative inotropic effect in isolated working heart in a dose-dependent manner. The negative inotropic of GSTT may be related to its action of blocking the calcium channel and has protective effect on lower glucosi cardiac.

Objective To study the effects of Gross Saponins of Tribulus Terrestris (GSTT) on thrombosis in vivo and in vitro. Methods ①The effects of GSTT on thrombus wet weight and thrombosis time of rats in vivo and the thrombus length, thrombus wet and dry weight in rats in vitro were observed by using the methods of neck artery-vein side-way thrombosis and artery thrombosis induced by electricity stimulation.②The effects of GSTT on the blood viscosity in blood stasis rats were observed. Results ①GSTT with doses of 20.8 and 31.2 mg•kg^-1 could significantly decrease thrombus wet weight (P<0.05 or P<0.01), and prolong thrombosis time (P<0.01), and shorten the thrombus length in vitro(P<0.05), and decrease thrombus wet and dry weight in vitro (P<0.01 or P<0.05). ②GSTT with doses of 20.8 and 31.2 mg•kg^-1 can significantly decrease blood viscosity (P<0.01). Conclusion GSTT has the effects of antithrombosis, and its mechanism is related to lowering blood viscosity and improving blood hemorheology.

Objective To study the effects of Gross Saponin of Tribulus Terrestris (GSTT) on acute myocardial infarction induced by ligating coronary artery in anaesthetic thoraco-opening dogs by injection. Methods Thirty domestic dogs were randomly divided into 5 groups (6 in each group): blanking control group, positive control group(Xuesaitong), GSTT groups with doses of 6.26 , 12.52, and 25.0 mg•kg^-1, respectively. The descending branch of left coronary artery was ligated to construct the model of acute myocardial infarction. The effects of GSTT on epicardium electrocardiogram(EECC), myocardial infarction range(MIS) , and serum myocardium enzyme were observed. Results The ischemia degree(Σ-ST, P<0.05) and the ischemia area(N-ST, P<0.05) were reduced in GSTT groups with various doses compared with controls. In addition, GSTT reduced the serum myocardium enzyme levels　(P<0.01) and myocardial infarction ranges(P<0.001) compared with controls. Conclusion GSTT can significantly protect ischemic myocardium of the anesthetic thoraco-opening dogs mediated by ligating coronary artery.

Objective To construct the prokaryotic expression vector of the fusion protein composed of protein transduction domain-eight arginine (8R) and MUC1 core peptides, and to express and purify the 8R-MUC1 core peptide fusion protein which should possess biological activity. Methods Two tandem repeat sequence of MUC1 core peptides were gotten from HSP65-MUC1-pET28a plasmid by PCR and cloned into pMD18-T vector. The T vector was digested by endonuclease for releasing the fragment of two repeats of MUC1 core peptides which was used for constructing six tandem repeat sequence of MUC1 core peptides by ligation. The fragment of six repeats of MUC1 core peptides was cloned into pET26b⁺ plasmid, which contained 8R, for constructing expression vector of the fusion protein composed of 8R and MUCPT (8R-MUCPT). The E.coli BL21(DE3) transformed with 8R-MUCPT-pET26b⁺ were induced by IPTG for expression of 8R-MUCPT fusion protein. The protein was purified by Ni²⁺ affinity chromatography. Results The prokaryotic expression vector, 8R-MUCPT-pET26b⁺, was constructed with 8R and six tandem repeat sequence of MUC1 core peptides. The fusion protein of 8R-MUCPT was expressed in BL21(DE3). The purity of the fu sion protein was 95.5% after purified by Ni²⁺ affinity chromatography. Conclusion The prokaryotic expression vector of 8R-MUC1 core peptide fusion protein has been constructed, and the fusion protein with biological activity has been successfully expressed and purified.

Objective To observe the glutathione S-transferas P (GST-P) expression in spontaneous hepatocellular carcinoma of LEC rats,then discuss the value of GST-P in early diagnosis for spontaneous hepatocellular carcinoma. Methods The immunohistochemical method was used to detect the expression of GST-P in LEC rat liver. Results The GST-P expression of hepatocarcinoma group of LEC rats (67 weeks) was higher than that in hepatitis group of LEC rat (4-6 months) and SD rat group (positive rates were 96.97%,37.5%,9.38%， respectively),（P<0.01）; the GST-P expression rate in hepatitis group of LEC rat (4-6 months) was higher than that in SD rat group（P<0.05). Conclusion GST-P can be regarded as the tumour marker to diagnose spontaneous hepatocellular carcinoma in early stage.

Objective To investigate the effects of prion protein 106-126 peptide on inducing apoptosis in differentiated PC12 cells. Methods After differentiated by nerve growth factor (NGF), the PC12 cells were infected by prion protein 106-126 peptide. Cell viability was determined by using the MTT assay and the morphological changes were observed. Apoptotic was detected by morphological changes, DNA electrophoresis and flow cytometry. Results Infected by this peptide, cell viability decreased 28.9%. Under light microscope, cells were shrinked and rounded, many cells were divorced from plate wall, some neuraxon shortened and broke. Apoptotic cells which nucleolus shrinked and rounded could be coloured orange by fluorescent colouration. Under electron microscope, chromatin gathered along the inside of the nuclear membrane, vacuole bodies appeared. Apoptotic peak was detected by flow cytometry and the ladder band appeared in DNA electrophoresis. Conclusion Prion protein 106-126 peptide can induce apoptosis in differentiated PC12 cells and present cellular toxicity definitely. It might be a perfect model to study the cellular toxicity of prion protein.

Objective To investigate the ameliorative effects of TangShenAn on renal functional and structural lesions in diabetic rats. Methods The Wistar rats were divided into four groups: control group(CN), diabetic group (DM), diabetic rats treated with TangShenAn (DM+T) and diabetic rats treated with aminoguanidine (DM+A). After TangShenAn （6 g•kg^-1）was administrated to the diabetic rats for 8 weeks, the renal function and structure were evaluated in the animals. Results As compared with controls, the levels of blood glucose, serum creatimine (SCr), blood urea nitrogen (BUN) and urinary protein excretion were significantly elevated in diabetic rats (P<0.01). After the diabetic rats were treated with TangShenAn, the blood glucose level was reduced, but there was no difference between TangShenAn-treated and untreated diabetic animals（P>0.05）, while the levels of SCr, BUN and urinary protein excretion were obviously decreased compared with untreated diabetic rats （P<0.01）. Administration of TangShenAn decreased the elevated glomerular volume/body weight in diabetic rats. Morphological examination of the renal tissue in diabetic rats exhibited an increased PAS-positive substance in mesangial areas, the segmental thickening of glomerular basement membrane and fusion of epithelial foot processes. After administration of TangShenAn to diabetic rats, these damages were markedly alleviated. Conclusion TangShenAn can ameliorate the renal functional and structural lesions in diabetic rats.

Objective To construct the complementary DNA (cDNA) of FⅧ-C2 by gene engineering and express the biologically active domain in NIH 3T3. Methods The total RNA extracted from 293 cells and FⅧ-C2 was amplified by RT-PCR. The product was then inserted into pTARGET vector together with signal peptide sequence. The recombinant vector was transfected into NIH3T3 cells followed by the detection of the expression of FⅧ-C2 by Western blotting. Results The cDNA of FⅧ-C2 was identified correctly by endonucleases cleavage and sequencing. The recombinant FⅧ-C2 peptide was obtained by detection of FⅧ-C2 from the supernatant of NIH3T3. Conclusion The construction of recombinant vector is a feasible way to express secretive FⅧ-C2.

Objective To study the expression and localization of s-lap/GFP fusion protein in Hela cell lines. Methods According to the coding area sequence of s-lap gene, stop codon located at 3′terminus of s-lap gene was mutated by PCR and fused with GFP gene. Then the s-lap/GFP fusion gene was transfected into Hela cells. The expression of s-lap/GFP fusion protein was observed under fluorescence microscope. Results DNA sequencing demonstrated that s-lap/GFP recombinant vector was constructed successfully and its fusion protein was expressed in Hela cells. The observation under fluorescence microscope showed that s-lap/GFP fusion protein was spread in entire cells, but expressed highly in the nucleus. Conclusion The s-lap/GFP fusion protein can express in Hela cells and locate in entire cells, but express highly in the nucleus.

Objective To explore the molecular mechanism of brain tissue injury induced by endotoxin through observation of LPO content, SOD activity, and ⅠκBα mRNA expression of cortex in endotoxic shock rats. Methods Wistar rats were randomly divided into lipopolysaccharide（LPS）, LPS+dexamethasone（Dex）, LPS+panaxadiol（PDS） and control group, respectively. The LPO content, SOD activity, and ⅠκBα mRNA expression were assayed 4 h after intravenous injection of LPS. Results MDA contents in LPS+Dex and LPS+PDS groups were obviously lower than that in LPS group (P<0.05), and the SOD activities were significantly higher than that in LPS group (P<0.05). Compared with the control group, ⅠκBα mRNA expressions in LPS+Dex and LPS+PDS groups were significantly higher than that in LPS group (P<0.05). Conclusion PDS may up-regulate ⅠκBα expression in the brain tissue, release the tissue peroxidation reaction induced by endotoxin and protect the central nervous system.

Objective Using a recombinant adenoviral vector carrying the human BMP-2 gene (Ad-BMP-2) to transfect rabbit bone marrow stromal cells (BMSC) to study the effects of Ad-BMP-2 transfection on cell differentiation. Methods The rabbit BMSC cultured in vitro were transfected by Ad-BMP-2. Seven days later, the expressions of BMP-2 and collagen type Ⅱ in these cells were determined by in situ hybridization and immunohistochemical analysis, and the alkaline phosphatase (AKP) activity of the BMSC was observed. Results There were positive results in cytoplasm in experimental group: the brown-yellow hybridization signals were observed and that indicated the expressions of BMP-2 and collagen type Ⅱ of Ad-BMP-2 infected BMSC by in situ hybridization, the yellow pigmenting were observed by immunohistochemical analysis but not in nuclei, the brown-black particles were seen by AKP staining. But in control group, these were negative. Conclusion Ad-BMP-2 could infect BMSC with high efficiency and promote the conversion to osteoblast and chondrocyte.

Objective To study the growth pattern and surface makers of adult human mesenchymal stem cells (hMSC) and the influences of bFGF on the proliferation of hMSC. Methods The growth curves of the hMSC were drawn; FACS was use d to detect surface antigenic markers. Cell growth pattern and its response to bFGF were evaluated by MTT test. Results The latent phase of the primary culture cells was 5-9 days, followed by 11-16 days of logarithmical proliferation; the plateau began at day 18-20. The subcultures showed 12-24 hours of latent phase, followed by 7-10 days of logarithmical proliferation, then entered the plateau (11-13 days). The hMSC expressed positive CD44 unanimously, but don′t expressed CD45. Low concentration of bFGF (10 μg•L^-1) could stimulate the proliferation of hMSC significantly (P<0.05); while 25-100 μg•L^-1 of bFGF could inhibit the ALP activity. Conclusion hMSC can be expanded in vitro rapidly. The effect of bFGF on hMSC proliferation is in positive dose-effect manner.

Objective To study the effects of sphingosine-1-phosphate(S1P) on calcium current of ventricular myocytes and its mechanisms. Methods The Langendorff perfusion method was used to isolate the ventricular myocytes of guinea pig and the ventricular myocytes of guinea pig were randomly divided into one control group, three S1P groups with dosages of 0.1, 1.0, and 10.0 μmol•L^-1, and group of PTX 0.1 mg•L^-1 together with S1P 1.0 μmol•L^-1. The whole cell voltage clamp technique was used to record the calcium current of isolated ventricular myocytes of guinea pig. Results S1P 1.0 μmol•L^-1 enchanced the calcium current from (0.256±0.030)mV before administration to (0.367±0.090)mV after administration (P<0.05). S1P 10.0 μmol•L^-1 enchanced the calcium current from (0.242±0.030)mV before administration to (0.364±0.060)mV after administration（P<0.01）. The effects of S1P to enchance the calcium current was blocked by PTX, a G protein-coupled receptor blocking agent. Conclusion S1P can promote the opening of calcium channels of the ventricular myocytes by G protein-coupled membrane receptor in a dose-dependence manner.

Objective To study the expression of connective tissue growth factor(CTGF)-RNA in mouse experimental hepatic fibrosis and its significance. Methods The liver tissue gene levels of CTGF were measured by reverse transcription polymerase chain reaction (RT-PCR) in 52 male Kunming mice with experimental hepatic fibrosis. Results The expression levels of CTGF were higher in hepatic fibrosis group than those in control group, they were closely associated with hepatic fibrosis development (r=0.582, P<0.01). Conclusion The mRNA expression of CTGF is remarkably up-regulated in mouse hepatic fibrosis. This result indicates that CTGF plays an important role in the activation and domination of hepatic fibrosis.

Objective To investigate the effects of Tripterium Regelii on urine protein and serum MMP-2, MMP9, and TGF-β₁ in nephritic rats. Methods The levels of serum MMP-2, MMP-9,and TGF-β₁ were detected by ELISA. Results The level of urine protein (mg/24 h) in nephritic rats treated with Tripterium Regelii was significantly decreased(pre-treatment: 327.38±48.67;post-treatment:170.38±32.62，P<0.01). The level of serum TGF-β₁ (ng•L^-1) was remarkably decreased too (model control: 287.60±43.37；model treatment:210.67±39.36，P<0.05)；the serum level of MMP-2 (μg•L^-1) was increased (model control: 240.63±38.34; model treatment:382.26±40.38，P<0.01)；the serum level of MMP-9 had a tendency to increase, but no significant difference was found (P>0.05). Conclusion Tripterium Regelii may have the effect of postponing glomerulosclerosis in rats.

Objective To study the ultrastructural changes of fibroblast apoptosis in the fracture repairing process. Methods The transmission electron microscope was used to observ the fibroblast proliferation and apoptosis in the fracture healing processes of rabbits. Results The results of the study indicated that the fibroblasts excreted collagens into the extracellular matrix, concomitant with formation of the numerous collagenous fibrils and diminution of cytoplasma, and the ultrastructual changes of fibroblasts eventually underwent apoptosis were found. Conclusion In the fibrous repairing processes, the morphologic changes of fibroblasts are characteristics of apoptosis.

Objective To explore the protective effects of oleanolic acid (OLA) derived from roots and leaves of Aralia Elata (Miq) Seem. on alcoholic liver disease (ALD) in rats and its mechanism. Methods Normal rats were intragastrically infused with ethanol and high fat diet. OLA was used from the beginning. Rats were sacrificed after 3 months interference. Hematoxylineosin (HE) method was used to examine the degree of hepatic injury. The concentrations of malandialdehyde (MDA), superoxide dismutase (SOD) and glutathione (GSH) were estimated in liver tissue. Blood samples were collected to measure the concentrations of aminotransferase, choline lipase (ChE), triglyceride (TG), total protein (TP), albumin (ALB), bilirubin (BIL), glutathione peroxidase (GSH-Px) and MDA. Results OLA decreased the levels of ALT, AST, TG, ChE, MDA in serum and the level of MDA in liver tissue in model group. In the same time it increased the level of GSH-Px in serum and the levels of SOD and GSH in liver tissue. Hepatic injury was reversed with OLA. Conclusion OLA can protect liver from alcohol injury in rats via eliminating free radicals and decreasing enzymes and blood fat in liver. Injury of free radicals and lipid oxidation are the main pathogenesis of alcoholic liver disease.

Objective To research the effects of extract of ginger on testis enzymes of X-ray irradiated mice. Methods Mice were treated with three different doses of extract of ginger:high dose (9.3 mL•kg^-1), middle dose (4.7 mL•kg^-1) ，and low dose (2.3 mL•kg^-1). All mice were irradiated once with 2.0 Gy X-ray. At the same time, the negative group (treated with vegetable oil only) and positive one (irradiated as well as extract of ginger groups after treated with vegetable oil) were set up. The changes of activities of enzymes in testes of mice were observed. Results After irradiated, in the group of high dose the activity of G-6-PD was decreased but the activity of LDH was increased (P<0.05); in the group of low dose, the activity of G-6-PD was increased while compared with control group, LDH had no significant difference (P>0.05). In every group, SDH had no significant difference (P>0.05). Conclusion The proper dose of extract of ginger has significant effects on stabilization of testis enzymes of X-ray irradiated mice.

Objective To investigate the anti-inflammatory effects of Hushigutong Granule. Methods Fifty Wistar rats were randomly divided into control group,aspirin group, and three groups of Hushigutong Granule at different doses (15,30,and 60 mg•kg^-1). Anti-inflammatory effects of Hushigutong Granule were carried through inflammatory models of auris swell induced by dimethylbenzene in mice, of foot swell induced by egg white and carageen glue in rats, of gasbag swell induced by croton oil in rats and penetration of capillary vessel. Results Hushigutong Granule (30 and 60 mg•kg^-1) had significant inhibitory action on auris swell induced by dimethylbenzene in mice, foot swell induced by egg white and carageen glue, gasbag swell induced by croton oil and penetration of capillary vessel in rats. There were significant differences compared with control group (P<0.05 or P<0.01). Conclusion Hushigutong Granule has significant inhibitory effects on many experimental inflammatory models.

Objective To study the influences of Niuhuanglingbei Xiaoyanpian (NHLBXYP) on both pigeon trcheocilia activities and rat tracheophenolsulfonphthalein excretion quantity. Methods The function of eliminating phlegm was determined by tracheocilia activity to motivate ink′s speed, the rat phenolsulfonphthalein excretion quantity was detected by phenolsulfonphthalein excretion. Results The ink′s moving distance was obviously extended in NHLBXYP groups with doses of 4,2, and 1 g•kg^-1, compared with control group (P<0.01). The tracheophenolsulfonphthalein excretion quantities in NHLBXYP groups with doses of 2.0,1.0,and 0.5 g•kg^-1 were significantly higher than control group (P<0.01). Conclusion NHLBXYP has the function of eliminating phlegm.

Objective To provide the applied anatomical fundamental data for extensive decompression during anterior cervical approach. Methods Forty-two adult aseptic cadavers whose arteries were affused with red latex were used for observing. The distances of the medial border of bilateral Longus colli muscle (A), the roots of anterior wall of transversal foramen(B), V2 segments of vertebral artery (C), between the medial borders of Longus colli muscle and the root of anterior wall of transversal foramen(D), between V2 segment of vertebral artery and internal wall of transversal foramen(E), between the medial border of superior cervical ganglion(F), middle cervical ganglion(G) and midline were measured. Results The lengths of A,B, and C gradually decreased from caudal location to cephalad location, but the lengths of F and G gradually increased; D was about 7.0 mm and E was about 3.0 mm. Conclusion The sympathetic trunk and vertebral artery injuries can be completely avoided if the root of anterior wall of transversal foramen as anatomical signal in extensive decompression during anterior cervical approach is considered.

Objective To research the applied anatomic characteristics of temporal limbus corneae and provide applied anatomic basis for incision of temporal limbus corneae. Methods The hierarchical structures of limbus corneae of 10 adult cadaveric eyeballs were observed under stereomicroscopy. The width and thickness of the temporal limbus corneae, the distance between main structures of the anterior chamber angle were measured. In addition, the comparison of 40 cases of uncontrolled intraocular pressure cases after the operation of glaucoma was used to prove the therapeutic efficacy of the incisions of temporal limbus corneae. Results By measuring the cadavers, the width of the temporal limbus corneae was (0.62±0.05)mm,the thickness was (0.73±0.04)mm. The distance between anterior border of the Schlemm′s duct and scleral process was (0.44±0.02)mm, the distance between anterior border of the Schlemm′s duct and the Schwalbe′s line was (0.24±0.03)mm. There was no obvious difference of sight between the incision of superior limbus corneae and the incision of temporal limbus corneae. The intraocular pressure of the incision of temporal limbus corneae was lowered than that of the incision of superior limbus corneae (P<0.05). Conclusion The incisions of temporal limbus corneae is applicable for the second operation of glaucoma and complicated cataract.

Objective To study and establish the culture system of thymic epithelial cell of BXSB systemic lupus erythematosus mice. Methods The thymic epithelial cells of systemic lupus erythematosus model (BXSB mice) were cultured with merely shearing, shearing and collagenase digestion, shearing and trypsinization in different nutrient media . The cells were identified with microscopy and immunohistochemical methods. Results The pure thymic epithelial cells were obtained not only in the medium with serum but also in the medium with growth factor. The thymic epithelial cells grew very well with shearing and collagenase digestion, and there was seldom contaminated by fibroblast. Passage was operated when the primary culture cells spreaded more than 90% pavement of culture flask. The purity of cells dyed with keratin was more than 95% in the medium with serum by shearing and collagenase digestion. Conclusion The method of shearing and collagenase digestion process is easy and cheap to obtain thymic epithelial cell of BXSB mice. A stable thymic epithelial cell culture system of BXSB systemic lupus erythematosus mice is established.

Objective To investigate the genetic association between the PTGS2/PLA2G4A gene polymorphism and schizophrenia in the Han descent population in the North of China. Methods A PCR-based RFLP procedure was employed to detect the genotypes of 168 family trios of Han descent population in the North of China. Results No significant differences were found in the frequencies of allele and genotype of the two SNPs (SNP1 and SNP2) of PTGS2 gene and one SNP(SNP3) of PLA2G4A gene between the case and the control(P>0.05). Conclusion There is no genetic association between the PTGS2/PLA2G4A gene polymorphism and schizophrenia in the Han descent population in the North of China.

Objective To compare the microvessel density (MVD) and the expressions of vascular endothelial growth factor (VEGF), CD44V⁶ and CD34 in ductal breast carcinoma between primary tumor and lymph node metastasis for researching the mechanism of metastasis and the corresponding reaction of body. Methods The expressions of CD34, VEGF, and CD44V⁶ were detected and analyzed in 30 cases of breast cancer with lymph node involvement (LN⁺) and 8 cases of breast cancer without lymph node involvement (LN^-) by immunohistochemical technique. Results Higher MVD was observed in the primary lesions than in the metastatic foci (P<0.05), and the expressions of VEGF were not significantly different between these two positions. There was a positive correlation between the expressions of VEGF and MVD in the primary tumor, and there was not a correlation between the expressions of VEGF and MVD in the metastatic foci in the lymph nodes. The expressions of CD44V⁶ were not significantly different between primary lesions and metastatic foci. The expressions of VEGF, CD34 and CD44V⁶ in the LN⁺ primary lesions were higher than those in the LN^- lesions (P<0.05). Conclusion The ability of angiogenesis between primary lesions of ductal breast carcinoma and its corresponding metastatic foci in the lymph nodes are different. Some elements in the metastatic foci can suppress the ability of angiogenesis.

Objective To study the expressions of matrix metalloproteinases-2 (MMP-2) in different grade and stage of transitional cell carcinoma of bladder（TCCB） and its significance. Methods Immunohistochemistry and in situ hybridization were used in 60 surgically resected specimens from patients with TCCB and 5 normal bladder tissues (used as control) to detect the expression of MMP-2. Results MMP-2 protein appeared to be far more abundant in stromal cells than tumor cells; MMP-2 mRNA appeared in stromal cells and tumor cells; in control group, the expression of MMP-2 was weak or negative; the positive expression of MMP-2 was significantly increased with stage and grade (P<0.01,respectively). Conclusion MMP-2 can express in TCCB and the expression of MMP-2 is significantly correlated with the aggressive and metastatic biologic behavior of TCCB.

Objective To study the expressions of p16, Bcl-2, Bax, VEGF and significance of microvessel density(MVD) in human pancreatic cancer. Methods Immunohistochemistry method and morphological semi-quantitative analysis were used to detect the quantity of MVD and the expressions of p16,Bcl-2,Bax, and VEGF in 33 cases of pancreatic cancer and 6 normal pancreas tissue specimen. Results The expression of P16 protein: it was higher in moderately or well differentiated pancreatic adenocarcinoma than poorly one (P<0.05), it was higher in patient with long survial time than those with short survial time (P<0.05);the expression of Bcl-2: it was higher in moderately or well differentiated pancreatic adenocarcinoma than poorly one; the expression of Bax:it was higher in normal pancreatic tissue than pancreatic cancer, as well as moderately or well differentiated than poorly one and survival time≥12 months than <12 months; the expression of VEGF:it was higher in patients with lymph node metastasis than without lymph node metastasis; MVD count: it was lower in normal pancreatic tissue than pancreatic cancer as well as with lymph node metastasis than without lymph node metastasis; the expression of MVD had positive correlation with VEGF expression （r=0.79，P<0.05）；p16 gene expression had negative correlation with VEGF expression and MVD （r=-0.69,P<0.05;r=-0.87,P<0.05），but Bcl-2expression had no correlation with other gene expressions. Bax expression had positive correlation with p16 gene expresion （r=0.54,P<0.05），and had negative correlation with VEGF expression and MVD（r=-0.66,P<0.05）. Conclusion The determination of p16, Bcl-2, Bax, VEGF expressions and MVD in human pancreatic cancer may be used to evaluate the prognosis and judge the differentiation of the adenocarcinoma.

Objective To study the association of polymorphism in TP53 gene with the susceptibility and radiation sensitivity of non-small-cell-lung cancer (NSCLC) of the population in the North of China. Methods Using RFLP-PCR assays, TP53 genotypes were detected by amplifying DNA fragments with sequence specific primers and digested by FnuDⅡ enzyme in 88 patients with NSCLC as well as 112 healthy controls. Results The C/C allele frequency was significantly higher in NSCLC patients than that in the healthy controls (χ²=5.65,P=0.017) . The C/C genotype frequency was significantly higher in NSCLC patients than that of the healthy controls (χ²=9.33,P=0.0023). The risk of C/C homozygotes in NSCLC patients was about 2.7 times against G/G homozygotes with odds ratio of 2.43 (95% CI=1.32-4.51). Conclusion In the population in the North of China, TP53 C/C genotype is closely associated with the susceptibility of NSCLC. There is no significant relationship between the polymorphism in TP53 gene and radiation sensitivity in NSCLC.

Objective To study the expression of IL-13 in T cell from peripheral blood of bronchial asthma patients and the effects of CTLA4-Ig on it in order to study further the pathogenesis of bronchial asthma and to search for a safty and effective route to treat bronchial asthma. Methods The methods of ELISA and RT-PCR were used to determine the expressions of IL-13 in T cell from periferal blood of asthma patients before and after CTLA4-Ig treatment. Results The expression of IL-13 in bronchial asthma patients was significantly higher than normal control group (P<0.01). CALA4-Ig inhibited remarkably the secretion of IL-13 (P<0.01). The expression of IL-13 couldn′t be obtained in normal control group with PHA stimulation, and it was increased in asthma group (P<0.01), and CTLA4-Ig could decrese the exprension of IL-13 obviously after PHA stimulation (P<0.01). Conclusion IL-13 can play an important role in the pathogenesis of bronchial asthma, CTLA4-Ig can inhilit the express of IL-13 in T cell and there is clinical significance for asthma treatment.

Objective To discuss the relationship between advanced glycation end produts (AGEs) and dyslipidemia and the onset of diabetic retinopathy (DR). Methods The serum AGEs and lipids levels were examined in 35 patients with type Ⅱ diabetes complicated with DR,31 patients with type Ⅱ diabetes without DR (NDR) and 16 normal subjects.Results ①The serum AGEs levels in DR group and NDR group were significantly higher than that in normal group (P<0.05). The serum AGEs level in DR group was higher than that in NDR group (P<0.05), but the difference was not significant. ②The serum LDL-C level in NDR group was remarkably higher compared with normal group (P<0.05), while the serum HDL-C and APOA1 levels in NDR group were significantly decreased compared with normal group (P<0.05). In DR group, the serum TCH,TG, and LDL-C levels were higher than those in normal group notably (P<0.05). ③The serum TCH, HDL-C, and LDL-C levels in DR group were significantly higher than those in NDR group (P<0.05). ④Linear correlative analysis showed that the serum AGEs level was positively related with the serum TCH, TG, LDL-C, and APOB levels（r values were 0.705, 0.51, 0.60,　and 0.60, respectively）. Conclusion The generation and accumulation of AGEs, dyslipidemia, as well as the interaction of them are important risk factors for the onset of DR in type Ⅱ diabetic patients. Strict control of blood glucose level and correction of dyslipidemia should be carried out simultaneously, so that the onset and progression of DR can be postponed and controlled more effectly.

Objective To study the clinical significance of detection of Ph¹ chromosome and bcr/abl fusion gene in the diagnosis for chronic granulocytic leukemia. Methods Ninety-five patients with chronic granulocytic leukemia and twenty patients with iron-deficiency anemia were selected. Bone marrow short-term culture was used to prepare the chromosome, bcr/abl fusion gene was analyzed by RT-PCR method. Results In 95 patients with chronic granulocytic leukemia,83(87.4%) cases were Ph¹ Chromosome positive,90(94.7%)cases were bcr/abl positive,7(7.4%)cases were Ph¹ negative and bcr/abl positive, 5(5.3%) cases were Ph¹ and bcr/abl negative. All 20 patients with iron-deficiency anemia were Ph¹ and bcr/abl negative. Conclusion The results indicated that the detection of Ph¹ chromosome combined with bcr/abl fusion gene could improve the diagnosis and differential diagnosis of chronic granulocytic leukemia.

Objective To explore the relations of recurrent spontaneous abortion (RSA) with anticardiolipin antibody (ACA) and antinuclear antibody (ANA). Methods The levels of ACA and ANA in serum of 98 RSA patients and 40 normal women were detected by ELISA. Results The positive rate of ACA in RSA patients (32.7%) was obviously higher than normal wowen (5.0%) (P<0.01);there was no difference of the positive rate of ACA in statistics between early and late RSA, as well as between 2 times and more than 2 times abortions of RSA patients(P>0.05); the positive rates of IgG and IgM were higher among IgM,IgG and IgA of ACA, only 4 RSA patients were positive in all, the positive rate of middle and high level of ACA was 37.5%(11/32), the rest were low levels. The positive rate of ANA in RSA patients (17.3%) was obviously higher than that in normal women (2.5%) (P<0.05). Conclusion ACA and ANA are related with RSA, and may be regarded as a immunological assistant diagnostic index.

Objective To investigate the defect in embedding of female inferitility, especially the abnormity of the endometrium acceptance. Methods Immunohistochemical method was used to test the expressions of extra cellar matrix proteins-collagen type Ⅳ (COLⅣ) and laminin (LN) on the secretory phase endometrium of 41 fertile women (fertility group)and 43 unexplained infertile women (infertility group). Results The expressions of COLⅣ and LN in fertility group were strong in early secretory phase, while their expressions were the strongest in mid-term secretory phase and the weakest in late secretory phase. The expressions of COLⅣ and LN in every secretory phase in the infertility group were stronger than that in the fertility group, the expressions of COLⅣ and LN in the secertory phase endometrium in the fertility group was different from that in the infertility group. The expressions of COLⅣ and LN in gland-epithelia of the endometrium and the basemembrance, the blood vessel endothelial basemembrance and the interstitial cells in the same group were different. Conclusion The COLⅣ and LN expressions in the secertory phase change periodically and participate in the function change of the endometrium and the embedding of the blastocyst. Their stronger expressions affect the identification,location, and adherence to the endometrium of marix,and lead to the failure of the blastocyst embedding.

Objective To study the expressions of p16 and p15 in human endometrial carcinoma，atypical hyperplasia of endometrium, and normal endometrium and its significances. Methods The expressions of p16 and p15 was examined in 57 specimens of malignant, 8 specimens of borderline,7 specimens of benign uterine endometrial tissues and 7 specimens of normal endometrium by immunohistochemistry S-P methods. Results The positive rates of p16 expression in the malignant and non-malignant tissues were 40.4% and 77.3%, respectively, significant difference was observed (P<0.05). The expression of p16 was associated with pathological types, pathological grades and operation-pathology stages, but no association was found with muscle infiltrations. The positive rates of p15 expression in the malignant and non-malignant tissues were 47.4% and 86.3%, respectively, significant difference was observed (P<0.05). The expression of p15 was associated with pathological grades, but no association was found with pathological types, muscle infiltrations, and operation-pathology stages. There was positive correlation between p16 expression and p15 expression in uterine endometrial carcinoma（r=0.91，P<0.05）. Conclusion Low expressions of p16 and p15 may contribute to the development of uterine endometrial carcinoma.

Objective To investigate the effects of nitric oxide(NO) and endothelin-1(ET-1) on the children with Henoch-Schonlein′s purpura(HSP). Methods The plasma NO and ET-1 levels of 48 children with HSP,inclnding 21 children with HSP nephritis (HSPN) and 27 children with non-HSPN (NHSPN), were determined both in acute and recovery phases. Results The concentrations of the plasma NO and ET-1 in acute phase were much higher than those in recovery phase and control group, respectively (P<0.01), and the changes were much more significant in children with HSPN than those without HSPN. The ratio of NO/ET-1 of hypertension group was lower than that of children without hypertension in acute phase (P<0.05). There were no significant differences of the plasma NO and ET-1 between NHSPN group in recovery phase and the control group（P>0.05）. However, the plasma NO and ET-1 concentrations of the HSPN group in recovery phase were still higher than those in the control group (P<0.05),and were lower than acute phase, and the ratio of NO/ET-1 in recovery phase was lower than that in acute phase. There was a positive correlation between NO and ET-1 in children with HSP (HSPN group in acute phase：r=0.7843，P<0.01,in recovery phase：r=0.4920，P<0.01)；NHSPN group in acute phase r=0.6831，P<0.01，in recovery phase r=0.0.6317，P<0.01）. Conclusion The levels of NO and ET-1 are important indexes to determine the state of illness, occurrence of complication and prognosis in children with HSP.

Objective To investigate the effects of methylation p16 on pathogenesis of actue lymphoblastic leukemia. Methods The restriction endonuclease enzyme digestion combined with PCR was used to detect p16 gene and methylation in 82 patients with acute lymphoblastic leukemia. Results One homozygous deletion was found in these cases and methylation of p16 gene was found in 38.3% patients (31/81) . Patients without methylation of p16 gene had a higher complete remission rate of first chemotherapy and a longer survival time than those with methylation of p16 gene. Conclusion Methylation of p16 gene probably plays a role in the pathogenesis of acute lymphoblastic leukemia and may have a prognostic value.

Objective To investigate the correlation of matrix metalloproteinase-2,-9 (MMP-2,MMP-9) in transitional cell carcinoma of bladder (TCCB) with tumor invasion and metastasis. Methods The total serum MMP-2 and MMP-9 levels were detected in 68 patients with TCCB and 20 healthy controls by ELISA; the expresions and distributions of MMP-2 and MMP-9 in 68 patients and 10 normal bladder tissue were determined by immunohistochemical method. Results The levels of serum MMP-2 and MMP-9 were significantly elevated in cancer samples compared with normal (P<0.01), significant difference was found between the patients with metastasis and without metastasis. In TCCB, the expressions of MMP-2 and MMP-9 were increased significantly compared with normal bladder epithelial. The expressions of MMP-2 and MMP-9 were correlated with histological grade and lymph node metastasis (P<0.05).　 Conclusion These results suggest that the total levels of serum MMP-2 and MMP-9 and their expressions may provide reliable information in TCCB prognosis.

Objective To study the changes of serum leptin in patients with simple obesity and patients with type 2 diabetes mellitus complicated with obesity in order to explore the relationship of leptin and insulin resistance and the role of leptin in the occurrence of type 2 diabetes mellitus. Methods 60 cases of simple obesity, 60 cases of type 2 diabetes mellitus and 30 cases of normal control were included according to the diagnostic criteria of obesity and type 2 diabetes mellitus. The levels of fasting serum leptin, fasting serum insulin, fasting glucose, fasting blood lipid were measured in all cases.The body mass index (BMI) and insulin action index were calculated. Results The level of BMI, serum leptin, serum insulin, blood lipid were significantly higher in patients with simple obesity and with type 2 diabetes mellitus complicated with obesity than in normal control cases, whilel (IAI) was significantly lower. The levels of free serum leptin, serum insulin, free glucose, and blood lipid were significantly higher in patients with type 2 diabetes mellitus complicated with obesity than in patients with simple obesity, while IAI was significantly lower. The level of serum leptin was positively correlated with BMI(r=0.48,P<0.55) and fasting serum leptin(r=0.55,P<0.05) and negatively correlated with IAI(r=-0.47,P<0.05)in patients with type 2 diabetes complicated with obesity. Conclusion The overexpression of serum leptin may play an important role in the occurrence of in insulin resistance and type 2 diabetes mellitus in obesity patients.

Objective To study the changes of serum vascular endothelial growth factor (VEGF) in patients with breast cancer and its significance. Methods The preoperative fasting serum VEGF levels were determined in 36 patients with breast cancer, 22 patients with breast fibroadenoma and breast cystic hyperplasia， and 35 healthy female controls by using ELISA. The relationships between VEGF and clinical indexes were analyzed. Results The serum VEGF level (ng•L^-1) in breast cancer group （145.8±13.6） was significantly higher than those in benign breast disease group （80.47±3.58） and control group （58.38±7.04） （P<0.05）. The VEGF level (ng•L^-1) in patients with lymph node metastasis （188.40±19.80） was higher than that in patients without lymph node metastasis （94.02±12.04）（P<0.01）. Conclusion The serum VEGF level has certain value to make differential diagnosis between malignant and benign breast diseases and predict the possibilities of prognosis and lymph node metastasis.

Objective To explore the clinical values of color kinesis (CK) combined with Doppler tissue imaging (DTI) in detection of regional wall movement abnormalities(RWMA) in patients with myocardial infarction. Methods The left ventricular wall movements were measured by CK and DTI techniques in 67 patients with myocardial infarction and 40 normal control subjects. DTI indexes included peak velocity of systolic wave(Vs), peak velocity of early or late diastolic wave(Ve, Va). Results The amplitudes of left ventricular wall movement by CK and the left ventricular wall movement velocities by DTI were decreased significantly in patients with myocardial infarction. The amplitudes and velocities of myocardial infarction segments were decreased further. The sensitivities of CK and DTI indexes were low in the diagnosis of RWMA (85.00% and 86.56%) and identification of infarction segments (87.62% and 85.33%) when they were used singly. When CK indexes were used with DTI indexes, the sensitivity of RWMA was 99.51%, the sensitiveity identification of infarction segments was 99.11%. Conclusion The RWMA and myocardial infarction segments can be diagnosed correctly by CK combined with DTI in patients with myocardial infarction.