Objective:To study the relationship between the T cell clonotypes and RA,carried out the analysis of infiltrating T cell clonotypes in foot of a murine model of spontaneous RA Methods:RT PCR and SSCP were used for this study Results:The identical rate of V?2 and V?8 2 clonotypes of T cells in different foot joints is 68 1 percent and 59 2 percent separately The assay of RT PCR/SSCP showed there were identical V?2 and V?8 2 clonotypes in four foot joints of all SKG mice(100%) DNA sequencing showed that the same DNA bands in samples from different joints by SSCP were the same T cell clonotypes The assay of T cell transformation suggested that clonotypes of T cells showed up in joints is related to morbid formation of RA Conclusion:This investigation indicated that T cells of V?2 and V?8 2 TCR types probably play an important role in SKG murine RA

ObjectiveTo study Decorin biological effects,Decorin gene cDNA was cloned and expressed in E. coli. MethodsThe Decorin gene obtained by PCR from T cell cDNA library and cloned into plasmid vector pUC19. The encoding sequence of Decorin in the pUC19 was confirmed by DNA sequencing using Sanger Dideoxy method, and then it was subcloned into expression plasmid vector pGEX-4T-1, the recombinant vector was indentified with BamHI and EcoRI in 1.2 ％ agarose gel electrophoresis. The expression of Decorin fusion protein with GST in E. coli JM109 was induced with IPTG and identified by SDS-PAGE. ResultsThe PCR amplified DNA fragment shared identical sequence with known Decorin gene reported in GenBank (Accession number: AF138300). The SDS-PAGE analysis revealed that the expressed fusion protein MW was approximatelly 65.6KD and in soluble format. ConclusionThe cloned Decorin gene is correct and it was expressed in fusion protein with GST.

Objective To study the expression and clinical significance of FoxM1 and Cep55 protein in basal-like breast carcinoma (BLBC).Methods The expression of FoxM1 and Cep55 protein in 66 cases of BLBC,70 cases of non-BLBC and 66 cases of normal adjacent breast tissue were detected by immunohistochemistry.Their relationship with the clinical pathological factors was analyzed.Results The positive rate of FoxM1 protein in BLBC,non-BLBC and normal adjacent breast tissue was 77.3％ (51/66),60.0％(42/70) and 13.6％(9/66),Cep55 protein was 74.2％(49/66),57.1％(40/70) and 16.7％(11/66),respectively.BLBC and non-BLBC tissue were higher than normal adjacent breast tissue,and BLBC tissue was higher than non-BLBC,there was significant difference (P ＜0.05).In BLBC tissue,the positive expression of FoxM1 and Cep55 protein correlated with lymph nodes metastasis and TNM staging (P ＜0.05),while did not correlate with age,menopause,tumor size (P ＞ 0.05).There was direct correlation between the positive expression of FoxM1 and Cep55 protein (r =0.259,P ＜ 0.05).Conclusion FoxM1 and Cep55 protein may play an important role in the carcinogenesis and development of BLBC,and both of them have the synergistic effect.

AIM: To apply the solid phase extraction (SPE) on salvianolic acids in Radix Salvia Miltiorrhizae by means of high performance liquid chromatography with mass spectroscopic detector (HPLC MSn). METHODS: A C 18 solid phase column and negative ion electrospray ionization (ESI) mode were used by orthogonal design to optimize four factors which might affect recoveries of samples, which included vacuum tightness, sample size, amount of eluting agent and eluting power. RESULTS: The optimized results were as follows: vacuum tightness was 0.002?106 Pa, sample size was 0.75 mL, amount of eluting agent was 15mL and eluting power was 1.0mL?s -1 . CONCLUSION: The established method is simple, reliable and suitable for the usual quality control method of salviamolic acids in Radix Salvia Miltiorrhizae.

Objective To detect the protein expressions of S100A7 and CD34 in basal-like breast carcinoma and analyze the clinical relationship between S100A7 and basal-like breast cancer and the connection with CD34 markers microvessel denisity (MVD).To study the function of S100A7 in basal-like breast cancer angiogenesis,invasion and metastasis.Methods Immunohistochemistry (IHC) was used to detect the expressions of S100A7 and CD34 in 60 cases of basal-like breast cancer,40 cases of non basal-like breast cancer and 20 cases of normal breast tissues.Results The expressions of S100A7 in basal-like breast cancer,non basallike breast cancer and normal breast tissues were 83.33％,62.50％ and 35.00％ respectively.There were significant differences among them (x2 =5.556,P ＜ 0.051; x2 =17.107,P ＜ 0.05; x2 =4.051,P ＜ 0.05).There was positive correlation between S100A7 and lymph node metastasis,TNM staging and MVD of basal-like breast cancer patients.But there was no significant correlation between S100A7 and patient's age and tumor sizes.Conclusion The up-regulation of S100A7 expressions may be associated with the occurrence and development,invasion and metastasis,angiogenesis of basal-like breast cancer.The effects of S100A7 promoting angiogenesis may be an important factor in the acceleration of the local invasion and distant metastasis in basallike breast cancer.

Objective To detect the protein expression of Cripto-1 and CD34 in basal-like breast carcinoma and study the functions of Cripto-1 in basal-like breast cancer angiogenesis,invasion and metastasis.Methods Immunohistochemistry (IHC) was used to detect the expression of Cripto-1 and CD34 in 60 cases of basal-like breast cancer,40 cases of non basal-like breast cancer and 20 cases of normal breast tissues.The clinical relationship between Cripto-1 and basal-like breast cancer were analyzed as well as its connection with CD34 markers MVD.Results The expression of Cripto-1 protein in basal-like breast cancer (42/60,70.00 ％) was significantly higher than that of nonbasal-like breast cancer (19/40,47.50 ％) and normal breast tissue (4/20,20.00 ％) (both P ＜ 0.05).The expression of Cripto-1 protein was positively correlated with the tumor size,clinical stage,lymph nodes metastasis and MVD value in patients of basal-like breast cancer (all P ＜ 0.05),but there was no corelation with the age of the patients (P ＞ 0.05).Conclusions The up-regulation of Cripto-1 expression may be associated with angiogenesis and development in basal-like breast cancer.The effect of Cripto-1 promoting angiogenesis may be an important factor in the acceleration of the local invasion and distant metastasis in basal-like breast cancer.

Aim To investigate the effects of monocrotaline pyrrole on production of nitric oxide and expression of eNOS protein of cultured pulmonary artery endothelial cells and on contractility of cultured pulmonary artery smooth muscle cells.Methods DAF-2 fluorescence technique was used to determine NO level,Western blot analysis was performed to determine the level of eNOS protein,and collagen gel contraction system was adopted to analyze muscle contractility.Results NO production induced by ACh and expression of eNOS protein were obviously inhibited by monocrotaline pyrrole compared with those of control group and gel contraction area in MCTP-treated cells induced by Thapsigargin obviously decreased.Conclusions monocrotaline pyrrole could inhibit the level of the ACh-induced production of NO and expression of eNOS protein,and enhance the contractility of pulmonary artery smooth muscle cells,which may be one of the possible mechanisms of MCTP-induced pulmonary artery hypertension.

Objective To study the biological characteristics of dendritic cells (DCs) derived from peripheral blood mononuclear cells (PBMCs) of patients diagnosed with syphilis. Methods PBMCs were isolated from 16 patients clinically and serologically diagnosed with syphilis, and from 16 healthy human controls, then cultured with GM-CSF and IL-4. On day 10, the monocyte-derived dendritic cells (MoDCs)of the patients and controls were collected and subjected to the detection of surface molecules by flow cytometry; TpN17 was used to stimulate MoDCs from the controls, the expression of phosphorylated ERK was detected by Westem blotting 20 minutes following the stimulation. Results The positivity rate of CD80 was significantly increased in the patients with syphilis than that in the controls (51.90% vs 33.67,P ＜ 0.05), while no significant difference was observed in the expressions of CD83, CD86 or HLA-DR be tween the two groups (16.53% vs 15.99%, 66.13% vs 59.32%, 91.29% vs 90.51%, all P 0.05). The ex pressions of CD80 and CD83 on the surface of MoDCs were enhanced in a dose-dependent manner after ex-posure to TpN17. The expression of cytoplasmic phosphorylated ERK was observed in MoDC stimulated by TpN17, but not in those without the treatment. Conclusions Antigenic stimulation with Treponema pal-lidum may be a reason for phenotypic abnormality of MoDCs derived from patients with syphilis. TpN17 may stimulate the maturation of DCs through the ERK signal transduction pathway.

Objective:To evaluate the clinical effect of laryngofissure without tracheotomy.Method:One hundred and sixteen cases of laryngofissure without tracheotomy in our department were analyzed retrospectively.Result:Of 116 cases, 101 cases had the laryngeal carcinoma; 9 cases had laryngeal stenosis; 6 cases had benign tumor of larynx. All the incisions were healed by first intention; and postoperative duration of hospital stay were 7 to 20 days. Three cases recurred in 101 cases of laryngeal carcinoma (2.97%), and underwent reoperation,the 5 year survival rate was 100%. One case recurred in 9 cases of laryngeal stenosis, and underwent reoperarion of laryngoplasty with T-tube implanted. No recurrence in 6 cases of benign tumor of larynx.Conclusion:Laryngofissure without tracheotomy have the advantage of less trauma , fast rehabilitation and no destroying the normal respiratory functions. It is completely feasible so long as surgeons grasp the adaption of operation strictly and select suitable cases.

Objective To investigate the effect of Dectin-1 on the release of inflammatory factors interleukin-10 (IL-10) and tumor necrosis factor-α (TNF-α) in rat tracheal epithelial cells (RTECs) stimulated with heat-treated Candida glabrata (C.glabrata).Methods RTECs cultivated in vitro were randomly divided into three groups,including control group(RTECs + sterile normal saline),fungal stimulation group(RTECs + heat-treated C.glabrata),and inhibitor intervention group(RTECs + laminarin + heat-treated C.glabrata),cells were harvested after incubation for 0,2,4,6 hours respectively,cell survival rate was determined by MTT method,expression of Dectin-1 was analyzed by Western Blot,expression of IL-10 and TNF-α were detected by enzyme-linked immunosorbent assay (ELISA).Results Heat-treated C.glabrata destroyed cell structure and reduced cell survival rate.At the beginning of the culture (0 h),cell survival rate,expressions of Dectin-1,IL-10 and TNF-α among three groups were all not significantly different(all P＞0.05).After incubation for 2,4,6 hours,expressions of Dectin-1,IL-10 and TNF-α in fungal stimulation group and inhibitor intervention group were both significantly higher than control group;expressions of Dectin-1,IL-10,and TNF-α in inhibitor intervention group was lower than fungal stimulation group(all P＜0.05).The expressions of IL-10 in inhibitor intervention group at 0 h and 2 h was not significantly different,expressions of Dectin-1,IL-10,and TNF-α in fungal stimulation group and inhibitor intervention group at different incubation periods were significantly different(all P＜0.05).Conclusion Dectin-1 is an important receptor for RTECs to recognize the heat-treated C.glabrata,it induces the release of IL-10 and TNF-α,and mediate the occurrence of inflammation.