Objective To study the effects of heat-shock protein (HSP) fusion protein on the differentiation and maturation of human monocyte derived dendritic cells (DC). Methods Human peripheral blood mononuclear cells (PBMC) were induced into immature dendritic cells (iDC) in the presences of GM-CSF and IL-4. On the 5th day the iDC was stimulated with HSP-fusion protein. Monocyte conditioned medium (MCM) and MCM mimic, a cocktail of IL-1β，IL-6，TNF-α and PGE2 were used as control. On the 7th day the DC stimulated with the HSP-fusion protein was stained with FITC-labeled antibodies against CD40, CD80, CD86, and HLA-A2 separately and the results were determined by FACS. Results HSP-fusion protein could up-regulate the iDC to express CD40, CD80, CD86, and HLA-A2 molecules. Conclusion HSP-fusion protein can induce the differentiation and maturation of human DC.

Objective To construct yeast expression vector which can express human glandular kallikrein highly. Methods The hK2 gene was amplified by RT-PCR from human normal prostate tissue. The amplified DNA fragment was cloned into pGEM-T vector for sequence analysis. The hK2 gene was then subcloned into yeast expression vector pPICZαC. Results A gene of 738bp was obtained and showed a 99% homology with hK2 gene sequence published in GenBank(NCBI: AF188746).Sequence analysis showed that only one amino acid was different.hK2 gene was inserted into yeast expression vector pPICZαC correctly. Conclusion The hK2 yeast expression vector pPICZαChK2 is constructed successfully in the study.

Objective To investigate the effects of advanced glycation end products (AGEs) on matrix metalloproteinase-2 (MMP-2) activity and expression in rat mesangial cells. Methods Rat mesangial cells were treated with 100 mg•L^-1 AGE-modified bovine serum albumin（AGEs group）or native bovine serum albumin（BSA group）for 48 h. Normal mesangial cells without any treatment were as a control (DMEM group). MMP-2 activity and mRNA were analyzed by zymography analysis and reverse transcriptase polymerase chain reaction (RT-PCR), respectively. Results Both MMP-2 activity and mRNA expression of mesangial cells in AGEs group were reduced significantly, compared with BSA or DMEM group (P<0.01). Conclusion AGEs can reduce the activity and expression of MMP-2 in rat mesangial cells.

Objective To explore the multipotency of rat bone marrow mesenchymal stem cells (rMSCs) in vitro. Methods rMSCs were obtained from rat bone marrow, cultured and passaged. rMSCs were induced to differentiate into chondrocytes in induction medium containing desamethasone, TGF-β1 and vitamin C. Then differentiated cells were identificated by toluidine blue stain and immunocytochemistry for collagen type Ⅱ. rMSCs were induced to differentiate into adipocytes in induction medium containing desamethasone and insulin. Then differentiated cells were identificated by Sudan IV stain. rMSCs were induced to differentiate into neuron-like cells in induction medium containing IBMX,GDNF， and IL-1β. Then neuron specific markers, such as NSE and MAP-2a,b, of the differentiated cells were identified by immunocytochemistry. Results rMSCs differentiated into chondrocytes on the 7th day after being induced. Toluidine blue metachromatic staining of differentiated chondrocytes in induction media was positive. Additionally, many differentiated chondrocytes in the experimental groups expressed collagen type Ⅱ. In vitro rMSCs differentiated into adipocytes and lipid droplets were scarlet red by Sudan Ⅳ staining on the 10th day after being induced. rMSCs differentiated into neuron-like cells and expressed NSE and MAP-2a,b by immunocytochemistry on the 7th day and the 15th day after being induced. The percentage of positive cells of NSE was (20.060±0.790)% and the percentage of positive cells of MAP-2a,b was (17.364±5.738)% on the 15th day.Conclusion rMSCs can differentiate into chondrocytes, adipocytes, and neuron-like cells in vitro, which shows that rMSCs have multipotency and are one of the best seeds cells in the tissue engineering.

Objective To explore the regulative mechanism of apoptosis-associated gene proteins on the adaptive response of thymocyte apoptosis in mice induced by low dose radiation with X-rays. Methods Kunming male mice were irradiated with the inductive doses (D1: 25, 50, 75, 100 and 200 mGy; dose rate: 12.5 mGy•min^-1) and the challenging dose (D2: 1.5 Gy; dose rate: 287 mGy•min^-1). The time interval between D1 and D2 was 6 h. The expressive levels of thymocyte apoptosis-associated gene proteins were measured with flow cytometry. Results As compared with the sham-irradiation, the positive percentage of thymocyte Bcl-2 protein expression decreased significantly in D2 group (P<0.05), Bax increased significantly (P<0.05), and Bcl-2/Bax decreased significantly (P<0.001); p53 increased significantly (P<0.001). As compared with D2 group, the positive percentage of thymocyte Bcl-2 protein expression increased in varying degree in D1+D2 group of 25~75 mGy D1, Bax decreased in varying degree, and Bcl-2/Bax increased significantly (P<0.01); p53 decreased significantly (P<0.001 or P<0.05). Conclusion The apoptotic thymocytes in the adaptive response of thymocyte apoptosis in mice induced by irradiation with 25~75 mGy decrease significantly due to the increase of apoptosis-associated gene Bcl-2 protein expression and Bcl-2/Bax, the decrease of Bax and p53 protein expressions.

Objective To explore the inhibitory effects of topotecan on mice with Lewis lung cancer(LLC) and its mechanisms. Methods LLC cells were inoculated subcutaneously into C57BL/6 mice. Fifty-five mice were randomly divided into control group (n=10) and experimental groups (Topotecan of 0.312,0.624， and 1.248 mg•kg^-1,15 in each group).Topotecan and phosphate- buffered saline vehicle were injected intraperitoneally, respectively.On the 21th day ,the mice were killed. Tumor growth curve was described, the inhibitory rate of primary tumor was measured. Microvessel density (MVD) was quantitated by immunohistochemistry using monoclonal antibodies against factor F8, vascular endothelial growth factor (VEGF) immunoreactivity was examined by immunohistochemistry using polyclonal antibodies against VEFG. Results Tumor growth curves in experimental groups were smooth compared with control group.There were significant differences of inhibitory rates of primary tumor between control and experimental groups(P<0.05);there were significant differences of MVD between high,middle dosages of topotecan groups and control (P<0.05).VEGF-positive rates were not significantly different between control and experimental groups. Conclusion Topotecan can effectively suppress the growth of the implanted LLC in C57BL/6 mouse and can inhibit angiogenesis.

Objective To clone the sequence of the cDNA of s-lap novel gene coding area and construct its expression vector. Methods The visible light damaged cultured retinal pigment epithelial (RPE) cell ds cDNA was used as template to obtain s-lap novel gene coding area sequence with RT-PCR and to construct its recombinant expression plasmid with recombinant DNA technique. Results Sequencing proved that the correct s-lap novel gene coding area sequence was obtained and s-lap gene coding area sequence was cloned to PHB expression vector by enzyme digestion identification and DNA sequence analysis，its recombinant expression vector was constructed successfully.Conclusion The s-lap novel gene coding area sequence is cloned and its recombinant expression plasmid PHB/s-lap is constructed successfully.

Objective To investigate the inhibitory effects of dehydroepaimdrosterone (DHEA) on the growth of transplanted Morris hepatomas(7288CTC) in vivo in rats and its mechanism. Methods 21 Buffalo rats were randomly devided into 4 groups, including one blank control (n=5), one group for tumor-bearing control (n=6), and 2 experimental groups with DHEA (n=6) or DHEA-s (n=4). DHEA or DHEA-s was fed to the rats for 4 weeks immediately after Morris hepatomas (7288CTC) was implanted in both flanks. Phenotypes of the spleen lymphocytes were examined by flow cytometry, PTEN expression in tumor cells was detected with specific MoAb in immunohistochemistry. The steroid contents in rat sera were determined by GC-MS. Results Tumor weight of DHEA treated group was less than those of controls (P<0.05), the inhibitory rate was 43%. The positive rate of PTEN protein in DHEA tumor group was higher than those in the controls. The percentage of CD3⁺ and CD4⁺ T cells in the spleens of DHEA-feeding rats increased significantly compared with the control group (P<0.05). There was not significant difference of serum steroid,TG and HDL levels between DHEA treated group and the control group. Conclusion The DHEA can inhibit tumor by increasing the PTEN protein expression and immune function.

Objective To investigate the fusion expression of staphylokinase（SAK） gene with (His)₆-tag and the purification of the expression product with Ni²+- metal chelating affinity chromatography. Methods The fragment of SAK cDNA was inserted into the pET-29b expression plasmid and the recombinant vector pET-29b-SAK was transformed into E. coli BL21(DE3) which was then induced by IPTG. The recombinant SAK-(His)₆ was purified with His•Tag affinity chromatography, and its bioactivity was analyzed. Results SDS-PAGE analysis indicated that the soluble fusion protein occupied 25% of the total soluble proteins in BL21(DE3). By one-step metal chelating chromatography using Ni-IDA resin, the purity of rSAK-(His)₆ was more than 90%. Then the recombinant target product obtained greater than 98% purity by the sequent exclusion chromatography using Sephacryl S-200HR. The specific activity of rSAK-(His)₆ purified was more than 5.0×10⁴ AU•mg^-1.　 Conclusion rSAK-(His)₆ has been expressed and purified successfully using pET-29b vector.

Objective To examine the inhibitory effects of Glycyrrhiza uralensis on sperm abnormalities induced by lead acetate,Pb(CH₃COO)₂, in mice. Methods Sixty-five mice were randomly divided into groups: negative control （N•S）, positive control（MMC 1.0 mg•kg^-1）， lead acetate mutagenesis groups（40，20, and 10 mg•kg^-1，which were matched 1/3，1/6, and 1/12 LD₅₀），Glycyrrhizia uralensis mutagenesis groups （1.25，2.50，5.00， and 10.00 g•kg^-1，which were matched 5×，10×，20×， and 40× clinical commonly used dosage of a person），Glycyrrhizia uralensis antimutagenesis groups （lead acetate 20 mg•kg^-1+ each Glycyrrhizia uralensis dosage groups）,5 mice per group. MMC and lead acetate were given by abdominal cavity injection, other drugs were given by perfusing stomach. Glycyrrhizia uralensis and lead acetate were given simultaneously in antimutagenesis groups,once a day for 5 days successively. At the 5th weekend after the first administration,the mice were killed by cervical vertebra dislocation whose epididymidis were drawed and sheets were produced in Wyrobek method.Results Sperm abnormality frequency (%) was no significant difference between tested groups（1.20±0.22，1.38±0.27，1.52±0.25，and 1.32±0.30）and control group（1.28±0.18） (P>0.05), but the frequency of abnormal sperms induced by lead acetate (%)（5.12±0.38） was reduced significantly by intragastrial perfusion of Glycyrrhiza uralensis and lead acetate （3.28±0.24，3.06±0.29，2.32±0.31， and 1.84±0.27）at the same time (P<0.01). Conclusion Glycyrrhiza uralensis can′t induce the sperm abnormality of mice,but shows a protective effect, namely antimutagenesis, on genetic materials in mice.

Objective To investigate the effects of high glucose Müller cell conditioned medium(HGMCM) on endothelial cells. Methods Immunofluorescence, flow cytometry and Boyden chamber migration models were used to observe the effects of HGMCM,wortmannin and VEGF monoclone antibody on endothelial cells. Results HGMCM could accelerate the proliferation and migration of endothelial cells (142.00±15.32/per sight), wortmannin (89.00±9.28/per sight) and anti-VEGF antibody (93.00±9.64/per sight) could inhibit the proliferation and migration of endothelial cells induced by HGMCM, and endothelial cells were blocked in G₁/S phase. There were significant difference compared with HGMCM group (P<0.01). Conclusion One of reasons of the proliferation and migration of endothelial cells induced by HGMCM is the role of VEGF. The anti-VEGF can inhibit the proliferation and migration of endothelial cells to inhibite angiogenesis.

Objective To study the influences of stationary magnetic fields on the contents of lipofuscin (LPF) in brain,heart, and kidney and the contents of GSH-Px in heart,kidney, and thymus of old mice. Methods Twenty female old mice were randomly divided into experimental group and control group. The mice in the experimental group were then put into a metal box where magnetic induction was 200 mT. They were exposed to the magnetic field one hour every day for 30 days running. Results The content of LPF in brain of old mice in experimental group ((0.160±0.024) μg•g^-1) was reduced remarkably (Ρ<0.05), compared with the control group ((0.304±0.061) μg•g^-1). But there were no significant differences of the contents of LPF in heart and kidney ((0.082±0.009) and (0.329± 0.024) μg•g^-1) (P>0.05) in experimental group, compared with the control group((0.092±0.011)and (0.382±0.033) μg•g^-1). The activities of GSH-Px in heart, kidney, and thymus of old mice in experimental group ((9.63±1.52),(30.68±4.50),and (96.72±10.94) NU•mL^-1) were increased remarkably (P<0.05 or P<0.01), compared with control group ((7.63±1.48),(24.09±4.49),and (85.68±7.122) NU•mL^-1). Conclusion In the stationary magnetic fields (20 mT), the contents of LPF in organs are reduced, free radicals are resisted and the activity of anti-oxide ferment is increased.

Objective To explain the effects of morphine on glutamate dehydrogenase (GLDH) and adenosine deaminase (ADA) in rat liver at a transcriptional level. Methods Fifty adult female Wistar rats, weighed （200±20）g， were randomly divided into 5 groups (10 in each group): control group ( injected saline for 7 days), morphine group Ⅰ and Ⅱ(injected morphine for 3 and 7 days， respectively), withdrawal group Ⅰ and Ⅱ (fed normally for 3 and 7 days separately following 7 days of morphine injection). The rat liver mRNA were extracted by RT-PCR, gene expressions of GLDH and ADA were analyzed after administration and after withdrawal. Results The GLDH mRNA contents in morphine group Ⅰ, Ⅱ and withdrawal group Ⅰ,Ⅱ were 1.02,1.09,1.16, and 1.46- fold of that in control group, respectively. The GLDH gene expression showed a continuing reinforcing trend after morphine injection compared with control group, even after drug withdrawal. The ADA mRNA contents in morphine group Ⅰ, Ⅱ and withdrawal group Ⅰ, Ⅱ were 2.65,1.89,1.18, and 0.87-fold of that in control group dividedly. The ADA gene expression increased after morphine injection and fell gradually after withdrawal. Conclusion Morphine can induce gene expressions of GLDH and ADA in rat liver.

Objective To obtain the best compatibility prescription to protect the stability of chromosome in which the Chinese herbal medicines had the role of anti-mutagesis including ginseng,slenderstyle acanthopanax root-bark, agaric, and Glycyrrhiza uralensis with orthogonal experiment. Methods The mutagen-sensibity experiment created by Hsu TC in which Bleomycin (BLM) was used as mutagen was adopted. The dosages of the above-mentioned Chinese herbal medicines in the experiment were applied in 3 levels: large dosage (double commonly used dosage), commonly used dosage and zero. The traditional Chinese medicines were randomly divided into four groups:A,B,C, and D. Ginseng was represented by C; slenderstyle acanthopanax root-bark was represented by D; agaric by A and Glycyrrhiza uralensis by B. The Chinese herbal medicines composed of different ratio with ginseng,slenderstyle acanthopanax root-bark, agaric, and Glycyrrhiza uralensis,respectively, were added into cultured lymphocytes of human peripheral blood.The chromosome breakage rates (b/c: chromosome breakage number per cell) intervened by the Chinese herbal medicines composed of different dosage ratio were compared and the anti-mutagesis effects were observed. Results The ranges of ginseng,slenderstyle acanthopanax root-bark ,agaric, and Glycyrrhiza uralensis was 16,14,9, and 6,respectively;the importance of the 4 kinds of Chinese herbal medicines in the compatibility prescription in order was ginseng,slenderstyle acanthopanax root-bark, agaric, and Glycyrrhizia uralensis; the best compatibility prescription was A₁B₂C₁D₂. Conclusion The best compatibility prescription to protect the stability of chromosome is A₁B₂C₁D₂.It is the greatest resistance to anti-mutagenic agents and reinforcement of chromosome stability to adopt above-mentioned dosage ratio.

Objective To research the factors influencing keratinocytes (KCs) proliferation activities of rat. Methods The KCs from rats were cultured with non-trophoblast technique in serum medium containing new-born calf serum. Epidermal growth factor(EGF), hepatocyte growth factor(HGF), fibroblast growth factor(FGF), choleratoxin(CT), hydrocortisone(HVB), insulin, and 3,5,3′-triiodothyronine (T₃) were added to the medium at the time of exponential phase of growth. The activities of cell proliferation were detected with MTT method. Results EGF, HGF, FGF, CT, HVB, insulin, and T₃ had promotion effects on KCs proliferation. Conclusion It is benefit for KCs culture to add some factors such as EGF, HGF, FGF, CT, HVB,insulin, and T₃ into medium.

Objective To study the effects of formaldehyde on lung histomorphology and the level of lipid peroxide in mice after inhaled different amount of formaldehyde. Methods Forty male mice were randomly divided into 4 groups: one control group and three formaldehyde dose groups(20，40, and 80 mg•m-3). The mice were poisoned 2 hours one day for 5 weeks continuously, then were killed. Lung tissues of mice inhaled formaldehyde were observed under light microscopy (LM). The activity of superoxide dismutase (SOD) and content of malondialdehyde (MDA) were detected biochemically in lung homogenate. Results It was manifestated as becoming bloodshots of pulmonary alveoli, expansion of pulmonary alveolus segment and inflammatory cells invasion under LM. The activities of SOD in 20,40, and 80 mg•m-3 dose groups were significantly lower than that in the control group, and the concentration of MDA was remarkably higher (P<0.01). The activity of SOD and the concentration of MDA had no remarkable differences between the different dose groups(P>0.05),but the former showed an increase activity in different dose groups, and the latter showed a reduced concentration in different dose groups. Conclusion Formaldehyde can injury free radical eliminated system to cause lung injury.

Objective To study the anatomic structure of suborbicularis oculi fat pad（SOFP）to guide the management of SOFP in the operation of removing the eyelid pouch and the plastic operation of lower eyelid in order to improve the curative effect of cosmesis. Methods The locations of SOFP in 48 corpse skulls (96 sides) were observed. The thickness of SOFP was measured. The comparisons of SOFP in different ages, sex, and somatotypes, between left side and right side in one corpse were made. Results The SOFP located the outside of orbit；among of suborbicularis oculi， top jaw bone and zygomatic bone′s periosteum. The fat pad′s appearance was irregular, connecting with each other, and with surroundings fat mat continuously, and the boundary was not clear. The SOFP′s thicknesses were 2.49-2.78 and 2.49-2.84 mm， respectively, in 20-30 and >30 years old (mean values were 2.66 and 2.68 mm, respectively). The SOFP′s thicknesses of males and females were 2.40-2.98 and 2.53-3.04 mm respectively (mean values were 2.75 and 2.79 mm, respectively）. The SOFP′s thicknesses of left side and right side were 2.53-2.65 and 2.51-2.69 mm（mean values were 2.59 and 2.61 mm, respectively ）. Therefore no significant differences of the SOFP′s thickness were found in different sex, ages or sides（P>0.05）. The SOFP′s thicknesses of emaciated, moderate, and obese somatotypes were 2.40-2.65,2.73-2.89, and 2.96-3.04 mm（mean values were 2.54, 2.81, and 2.99 mm, respectively）.So there were significant differences of SOFP′s thickness among different somatotypes (P<0.05). Conclusion In the plastic operation of eyelid pouch, the SOFP should be cut off at the same time, or reply the mistaken fat pad to improve the curative effect.

Objective To investigate the relationship between the expressions of aquaporin 7 (AQP7) and aquaporin 8(AQP8) and the spermatogenesis of rat testis, and the effects of panaxadiol saponins(PDS) on expressions of AQP7 and AQP8. Methods Wistar rats of 4 and 12 weeks old were divided into saline control (Con group) and PDS groups. The rats in PDS group were injected with PDS in abdominal cavity (25 mg•kg^-1•d^-1，1 mL) every day, and saline was used in Con group with equal volume for 2 weeks. Semi-quantitative RT-PCR and Western blotting were employed to detect the mRNA and protein expression levels of AQP7 and AQP8. Results Expression levels of AQP7 and AQP8 were elevated when the age of rats increased; two weeks after injection of PDS, the expressions of AQP7 and AQP8 were upregulated in 4 weeks old rats, but the expressions of AQP7 and AQP8 in 12 weeks old rats were inhibited. Conclusion The expression levels of AQP7 and AQP8 are increased significantly with the sexual maturation; PDS can regulate the expressions of AQPs in testis.

Objective To explore the effects of melatonin (MLT) on the damage of mouse thymocytes in vivo induced by ionizing radiation and its mechanism. Methods The exogenous MLT was given to Kunming mice to establish the animal models of single and successive administration of MLT through intraperitoneal injection before whole-body irradiation with 1 Gy X-rays. For single administration of MLT, the apoptotic body percentage(ABP) and DNA lytic rate(DLR) in the thymocytes were determined with flow cytometry and fluorospectrophotometry, respectively, 12 h after irradiation. For successive administration of MLT, ³H-TdR incorporative rate (HTIR) was determined 24 h after irradiation. Results The number of thymocytes in single adminstration group was significantly lower than that in the sham-irradiation group 12 h after irradiation with 1 Gy X-rays (P<0.001), while ABP and DLR increased significantly (P<0.001). When single administration of exogenous MLT was given before irradiation, the number of thymocytes in the 0.5 mg•kg^-1 MLT group was significantly higher, while the ABP and DLR were significantly lower than those in 0 mg•kg^-1 MLT group (simple irradiation, P<0.01 or P<0.001). The number of thymocytes and HTIR in adminsitration groups 24 h after irradiation were significantly lower than those in the sham-irradiation group (P<0.001). When successive administration of MLT was given for 1 week before irradiation, the numbers of thymocytes in adminsitration groups with 0.01~0.1 mg•kg^-1 MLT were significantly higher than that in 0 mg•kg^-1 MLT group (P<0.05), and the HTIR in 0.1~1.0 mg•kg^-1 MLT group was also significantly higher (P<0.05). Conclusion The administration of exogenous MLT before irradiation can decrease the damage of mouse thymocytes induced by ionizing radiation, and has the protective effect on immune functions in mice.

Objective To study the effects of CD8⁺ T lymphocytes on processing macrophages (MΦ) of patients with asthma. Methods The MΦ and CD8⁺ T lymphocytes were isolated from peripheral blood of 20 patients with asthma and 22 health donors, respectively. Every sample was divided into four groups. All groups were stimulated with CTLL₂P antigen for 18 hours and CTLL₂P antigen were washed away, then they were cultured with B lymphocytes together. The levels of secreted CTLL₂P-specific IgM, IgG, and IgE in the supernatant were measured after 10 days. Results ①When MΦ processed antigen only, the level of CTLL₂P-specific IgM of patients with asthma（0.034±0.022） was lower than that of health donors (0.116±0.080) (P<0.05). The level of CTLL₂P-specific IgM of patients with asthma (0.031±0.021) was lower than that of health donors (0.079±0.064) (P<0.05)，when CD8⁺ T lymphocytes were cultured with mononuclear phagocytes together. ②When CD8⁺ T lymphocytes were cultured with mononuclear phagocytes together, the level of CTLL₂P-specific IgG of patients with asthma (0.102±0.041) was significantly higher than that of health donors (0.081±0.067) (P<0.05). However, the IgG levels had not differences when active CD8⁺ T lymphocytes or natural CD8⁺ T lymphocytes were cultured with antigen processing mononuclear phagocytes (0.105±0.066，0.079±0.059)，compared with health donors (0.066±0.038，0.069±0.047)(P>0.05). ③The CTLL₂P-specific IgE level of patients with asthma (0.171±0.154) was higher than that of health donors (0.147±0.059) (P<0.05), when CD8⁺ T lymphocytes were cultured with mononuclear phagocytes together to process antigen. Conclusion The CD8⁺ T lymphocytes of patients with asthma have different regulatory effect on immunolglobulin production from antigen processed by MΦ.

Objective To study the association of single nucleotide polymorphisms (SNPs) in the estrogen receptor alpha gene(ESR1) with breast cancer susceptibility and different tumor clinical pathological characteristics. Methods The PCR-RFLP method was used to examine ESR1 genotype in 193 women with breast cancer and 71 healthy women. The potential role of the ER alpha genotype in estrogen receptor (ER), progesterone receptor (PR), cerbB2 expression and some other clinical pathological manifestations were assessed by SPSS 11.0 software. Results ①The allele frequencies of RS2077647 accorded with Hardy-Weinberg equilibrium;②The allele and the genotype frequencies of RS2077647 were positively associated with the age of patient, the size of the tumor and PR expression. The distributions of allele and genotype frequencies of RS2077647 were statistically different in breast cancer patients sub-groups classified by age, tumor size and PR expression （P＜0.05);③The distributions of RS2077647 genotype had no significant difference between the patients and healthy women. Conclusion RS2077647 polymorphisms is positively associated with the age of the patient, the size of the tumor and PR expression. It has no relationship with breast cancer susceptibility.

Objective To study the relationship between donor-derived NK cells and hematopoietic reconstitution after nonmyeloablative MHC haplotype-mismatched BMT in mice. Methods The murine model of MHC haplotype-mismatched BMT was established by using (BALB/c^H-2d×C57BL／6^H-2b) CB6F₁^H-2d/b mouse as the recipient, and C57BL／6^H-2b mouse as the donor. 60 recipient mice were divided into 10 groups after irradiation (TBI ⁶⁰Co 9.0,7.0， and 6.0 Gy)：9.0 Gy-irradiation groups including simple-irradiation group (Ⅰ), MHC haplotype-mismatched BMT group(Ⅱ); 7.0 Gy-irradiation groups including simple-irradiation group (Ⅲ), MHC haplotype-mismatched BMT group (Ⅳ), the first of NK cells-transplantation group (Ⅴ), the second of NK cells-transplantation group (Ⅵ), 6.0 Gy-irradiation groups including simple-irradiation group (Ⅶ), MHC haplotype-mismatched BMT group (Ⅷ), the first of NK cells-transplantation group (Ⅸ), the second of NK cells-transplantation group (X). The recipients were infused into the bone marrow cells of donors when they were irradiated after 4 hours. The recipients, the first of NK cells-transplantation groups, were infused into NK cells of 1×10⁶/one; The recipients, the second of NK cells-transplantation groups, were infused into NK cells of 5×10⁵/one. The effects were assessed by the count of WBC and PLT, survival time, body weight in the recipients. Results ①Life span: survival time was (5.83±0.98) days in group Ⅰ, beyond 60 days in other groups. ②Leukocyte and platelet counts after 10 days: leukocyte and platelet counts were (1.05±0.30)×10⁹• L^-1,（53.50±12.11）×10⁹•L^-1 and （0.95±0.26）×10⁹•L^-1,（47.17±11.82）×10⁹•L^-1 in group Ⅴ and group Ⅵ, which were much higher than those in group Ⅲ and group Ⅳ (P<0.01); In the cases of group Ⅸ and group X, leukocyte and platelet counts were （0.98±0.34）×10⁹•L^-1,（50.67±12.58）×10⁹•L^-1;（0.88±0.29）×10⁹•L^-1,（45.33±14.36）×10⁹•L^-1, which were much higher than those in group Ⅶ and group Ⅷ (P<0.05). Conclusion Donor-derived NK cells can promote hematopoietic reconstitution after nonmyeloablative MHC haplotype-mismatched BMT in mice.

Objective To induce the immune tolerance of recipient to donor trachea pretreated with immunosuppressor. Methods Twenty adult dogs were used, 4 dogs were donors, the others were recipients. The donor dogs were killed and all tracheas were divided into 16 segments. The 2 donor dogs with 8 tracheal segments pretreated by immunosuppressor （CTX 750 mg and MPED 2 g） were used as experimental group, the other 2 donor dogs with 8 tracheal segments without immunosuppressor pretreatment as control group. After the tracheal segments were embedded with omentum in the recipient peritoneal cavity for 3 weeks, all tracheal segments were taken out to evaluate the scale of immune reaction pathologically. Results The normal histological construction was kept in 7 tracheal segments except 1 segment with severe immune rejection in the experimental group. 3 segments with mild rejection, other 3 segments with moderate rejection and another 2 tracheal segments with severe rejection were found in the control group. There was no significant difference between the two groups (P>0.05). Conclusion The pretreatment of donor with immunosuppressor can induce recipient immune tolerance for the canine tracheal allotransplantation.

Objective To investigate the expressions of matrix metalloproteinase-2（MMP-2, gelatinase A）and its inhibitor，tissue inhibitor of metalloproteinase-2（TIMP-2），in human glioma. Methods The samples from 35 cases of glioma were divided into four groups according to the WHO classification: grade Ⅰ (n=7), grade Ⅱ (n=12), grade Ⅲ (n=10)， and grade Ⅳ (n=6), 4 normal brain samples were served as the control. The expressions of MMP-2 and TIMP-2 protein were determined with immunohistochemistry in the samples, and the correlation between the expressions of MMP-2, TIMP-2 and malignant degree of the gliomas was analyzed. Results The positive staining was localized in the cytoplasm of glioma cells and extracellular matrix. In the gliomas from gradeⅠ to gradeⅣ, the positive staining rates (%)(10.42±3.95，29.92±7.69，45.40±9.01， and 57.17±11.39, respectively) and the expression intensities (2.14±0.38，3.17±0.71，4.00±0.70, and 5.83±0.41, respectively) of MMP-2 were significantly elevated with the malignant grade of the gliomas, while the positive staining rates (%)(51.81±12.28，36.42±9.68，22.10±7.71， and 4.33±1.63, respectively) and immunostaining intensities (5.57±0.53，3.92±0.29，2.70±0.50， and 2.17±0.41, respectively) of TIMP-2 were markedly reduced with increasing the grade of glioma malignancy. The ratioes of MMP-2/TIMP-2 were obviously increased when the malignant grades of the gliomas were elevated. There was a negative correlation　(r＝－0.69，P<0.05) between MMP-2 and TIMP-2 expressions in the gliomas. Conclusion MMP-2 and TIMP-2 together play an important role in invasiveness of human glioma.

Objective To explore the roles of abnormal protein expression of cluster of differentiation (CD44v6) and mRNA expressions of α- and β-catenin in metastasis of laryngeal cancer. Methods CD44v6 protein expressions and mRNA expressions of α- and β-catenin in 64 cases of laryngeal cancer and 9 cases of normal laryngeal tissues were detected with the methods of immunohistochemistry and RT-PCR. Results Positive protein expressions of CD44v6（Gray level：173.1±15.0）were observed in normal controls，and negative expressions（9 cases，Gray level：>200）or low expressions（14 caces，Gray level：186-200）of CD44v6 were found in the laryngeal cancer tissues. Abnormal expression of CD44v6 protein was correlated with histological grades and lymph node metastasis in laryngeal cancer. Abnormal expressions of α- and β-catenin mRNA were detected in 6 samples (negative:4 cases; deletion of 700 bp gene fragment:2 cases) and 10 samples (negative:7 cases; deletion of 350 bp gene fragment:3 cases) with laryngeal cancer, respectively. Abnormal expressions of α-catenin and β-catenin mRNA were apparently associated with clinical stages and lymph node metastasis in laryngeal cancer (P<0.05). Conclusion Both negative or low expressions of CD44v6 and abnormal mRNA expressions of α-catenin and β-catenin can play a role in the lymph node metastasis of laryngeal cancer.

Objective To study the effects of Utilin′s on the immune function and the pleural effusion absorption in patients with tuberculous pleurisy treated with antituberculotics. Methods The 43 tuberculous pleurisy patients were randomly divided into two groups, one group was treated with antituberculotics only (control group, n=23), the other group was treated with antituberculotics and Utilin′s (experimental group, n=20). The treatment duration was half year. The pleural effusion absorption was observed and the activities of IgG, IgA, IgM, CD3⁺, CD4⁺, CD8⁺ and NK of the 43 patients were measured before and after treament. Results ①In three months, the numbers in experimental group whose pleural effusion were absorped wholely were more than that in control group, and the difference was significant four months after treatment (P<0.05); ②Half year later, the levels of CD3⁺ and CD4⁺ in experimental group were significantly higher than those in control group (P<0.01), and NK cell activity in experimental group was significantly higher than that in control group (P<0.05), and the IgG level was increased (P<0.05). Conclusion Utilin′s can stimulate the immune system of tuberculous pleurisy patients and accelerate the pleural effusion absorption.

Objective To observe the TOROH(including CMV,RV,TOX,and HSV) infections of newborns in hospital and study the changes of the levels of IL-4, IFN-γ, and TNF-α, respectively,in serum. Methods The neonates were given serum TORCH-IgM tests by ELISA.The levels of IL-4, IFN-γ, and TNF-α were all detected by double antibody with filling ELISA. Results There were altogether 18 patients with positive TORCH-IgM of 65 neonates in hospital. Among them there were 14 patients with positive CMV-IgM, 8 with TOX-IgM, 5 with positive HSV₂-IgM, and 3 with positive RV-IgM. The infect rates were 21.54%,12.31%,7.69%, and 4.62%, respectively,among neonates in hospital at the same time .The levels of IL-4, IFN-γ, and TNF-α in patients with positive TORCH-IgM were 20.47±2.55,292.6±73.9,and 219.11±22.27 ng•L^-1,respectively, while those in normal control group were 27.11±8.97,204.5±25.7, and 147.74±20.19　ng•L^-1,respectively. The level of IL-4 of patients with positive TORCH-IgM decreased (P<0.05),the levels of IFN-γ and TNF-α increased (P<0.05),compared with control group.　 Conclusion TORCH infection is very common in neonates in hospital. The detection rate of positive CMV-IgM is the highest, TOX-IgM comes next. After TORCH infection, the balance of immune is destroyed, and cytokines are secreted imbalanced. In the TORCH infection treatment, the immunity adjusting treatment should be emphasized besides anti-pathogen therapy.

Objective To study the expression of platelet membrane glycoproteins (MG) CD62P in peripheral blood in patients with transient ischemic attack (TIA) and its clinical significance, and the effects of aspirin on platelet MG. Methods The CD62P expressions of 30 cases of TIA patients were measured with flow cytometry (FCM), including patients (9 cases) with progressing to infarction and non-progressing (21 cases). Whether taking aspirin orally or not, samples were divided into 2 groups (the former 12 cases, the latter 18 cases). And the results between groups and between the patients and healthy volunteers were compared. Results The expression of CD62P in healthy volunteers was lower, the positive rate was (30.17±3.55)% while that of TIA patients was higher, the positive rate was (65.23±21.11)%, among which positive rate of patients progressing to infarction was (68.21±2.42)% and that of non-progressing patients was (60.07±3.46)%. After TIA patients took orally aspirin more than 6 months regularly, the activation rate of platelet was (60.75±2.03)%. Patients without aspirin taken orally had a platelet activation rate of (70.01±3.43)%. Conclusion CD62P is able to reflect the activation level of platelet so as to forecast the onset of TIA and to estimate its evolvement trend; aspirin may play a suppressive effect on CD62P expression.

Objective To investigate the effects of He-Ne laser irradiation on VEGF expression in periodontium of experimental tooth movement in rabbits. Methods Thirty rabbits were randomly divided into 6 groups on average: forced group of 1, 3, 5, 7, 14, 21 days. An orthodontic appliance, consisting of a coil spring ligated to the first molar connecting it to an orthodontic wire ligated onto the incisors, and exerting a force of approximately 0.08 N. The right side was designed as experimental side, left side as control side, in the self-control. Experimental side received irradiation of He-Ne laser. The animals from each group were killed at the time discontinued. The histological sections were stained with mouse anti-rabbit VEGF monoclonal antibody to express VEGF immunoreactivity, using a SP KIT, and counterstained with hematoxylin. Results The VEGF expression of irridiation side in forced group of 1 day （130.02±4.29） was significantly lower than that in control side （136.74±4.56）（P<0.05） in the pressure region,but the VEGF expressions of irridiation sides in forced groups of 3 days and 5 days （151.20±2.98 and 162.11±4.32） were significantly higher than those in control sides （142.38±4.87 and 151.10±6.45）（P<0.05） in the pressure regions. The VEGF expressions of irridiation sides of forced group of 3,5, and 7 days （143.96±3.63,148.40±5.79, and 150.72±5.79） were remarkably increased, compared with control sides （138.20±4.88,138.36±6.17, and 140.20±6.35）（P<0.05） in the tension regions. Conclusion He-Ne laser irradiation can upregulate the expression of VEGF in periodontium of experimental tooth movement in rabbits.

Objective To evaluate the security of the treatment of paroxysmal supraventricular tachycardia (PSVT) with radiofrequency catheter ablation (RFCA), and observe the effect of radiofrequency energy on cardiac muscle. Methods 216 patients with PSVT (male 112, female 104) ,mean age (43.3±14.0) years old, were included. RFCA was performed in all patients with atrioventricular nodal reentrant tachycardia （AVNRT） or atrioventricular reentrant tachycardia （AVRT） proved by electrophysiological testing. The success rate, recurrence rate and complications were investigated. The contents of cardiac troponin T(cTnT) in patients before operations and 2 hours after operation were measured. Results The success rates of AVRT and AVNRT patients were 97.1%（134/138） and 98.7%（77/78）,respectively. The total success rate was 97.6%(211/216). The recurrence rates of AVRT and AVNRT patients were 3.6% and 5.1%, respectively. The total recurrence rates was 4.16%. Complications occurred in 5 patients (1.8%). The concentration of cTnT（0.850±0.083 μg•L^-1）in 216 patients 2 h after operation significantly increased compared with that in patients before operation （0.132±0.002 μg•L^-1）. The concentration of cTnT in 115 patients of twice and more twice discharged （1.200±0.091 μg•L^-1） significantly increased compared with that in 101 patients of once discharged 2 hours after operation （0.630±0.142 μg•L^-1）. Conclusion RFCA is an effective and safe method for PSVT.

Objective To investigate the effects of adenosine triphosphate-sensitive potassium (K⁺_ATP) channel opener nicorandil (Nic), K⁺_ATP inhibitor glibenclamide (Gli) and voltage gated potassium (K⁺_V) channel inhibitor 4-aminopyridine (4-AP) on the morphine (Mor) antinociception during intrathecal administration in rats. Methods Chronic neuropathic model was set up by ligation of left L₅ spinal nerve of Sprague-Dawley rat. Three days later rat was implanted catheter (PE-10) intrathecally. The thermal nociceptive threshold was determined by measuring the withdrawal latency of hindpaw placed on a 58℃ hot plate 7 days after operation. Results Pain threshold of rat′s left hindpaw of ligature group measured by heat stimulation was significantly lowered than that in control group 7 days after operation (P<0.01). The intrathecal administration of Nic 5 μg, Gli 2 μg, 4-AP 2 μg and saline alone did not produce any antinociceptive action, but Nic 5 μg did potentiate the antinociceptive effect induced by intrathecal morphine, and Gli 2 μg abolished the antinociceptive effect induced by intrathecal morphine (P<0.05), whereas 4-AP did not affect the analgesia of morphine. Conclusion K⁺_ATP channels can regulate the analgesic actions of morphine at the spinal cord level.

Objective To discovery the stress distribution in bone around the implant of middle implant-teeth fixed bridge under different loading. Methods The model was set up by 3-D finite element means. The stress in bone around the implant under loading was computed by finite element analysis software Super-SAP 93. Results The biggest stress in bone around the implant lied in the neck cortex of bone. The biggest stress in bone around the implant under concentrated oblique loading was 2.5 times as big as that under concentrated vertical loading. The peak value of tensile stress lied in the boundary of lingual cortex of bone under concentrated vertical loading. The biggest compressive stress was higher than the biggest tensile stress under concentrated loading. Conclusion It is important not only to absent abnormally high occlusal points, but also to reduce side direction force for arrangement of natural teeth, middle implant, and fixed bridge into balanced occlusion.

Objective To study the relationship between West syndrome and multidrug resistance gene (MDR). Methods The expressive levels of perihemo-lymphcyte membrance glycoprotein (P170) in patients with West syndrome（n=12），partial seizures (n=9)，generalized seizures (n=8), and 8 healthy children were detected by flow cytometry. Results The expressive levels of perihemo- glycoprotein 170(P170) increased in patients with common epilepsy and West syndrome, compared with the healthy group;the expressive level of P170 in patients with West syndrome (59.52%) was strongly higher than that in the control group (1.22%)(P<0.001) and common epilepsy group (11.47%)(P<0.05). Conclusion The increasing of the expressive level of multidrug resistance gene P170 may be the important cause of difficulty in therapy of West syndrome.