Objective To investigate the effects of cadmium on bone loss in ovariectomized rats. Methods Female Sprague-Dawley rats, aged 6 months, were divided to 5 groups, i.e. sham operation group (sh) , control group (ovariectomy 0 mg/L Cd), low dose group (ovariectomy+50 mg/L Cd), medium dose group (ovariectomy+100 mg/L Cd), high dose group (ovariectomy+200 mg/L Cd). Cadmium was administrated via drinking water. After 24- week exposure, the following indices were measured, bone mineral density (BMD) at the femurs necks, serum estradiol, urinary and blood cadmium, urinary ? 2-MG, bone calcium and cadmium, and calcium contents in feces and urine. Results Estradiol and BMD significantly declined in the ovariectomized rats. Cadmium administration aggravated the declines of BMD in a dose-effect pattern. Urinary ? 2-MG, the excretions of calcium in feces and urine were significantly higher in cadmium-exposed groups in a dose-effects pattern. And bone calcium was also significantly lower in the group exposed to 200 mg/L cadmium than in the control group. Conclusion It is indicated that cadmium could increase bone loss and decrease BMD in ovariectomized rats by increasing the excretions of calcium in feces and urine.

Objective　To investigate the activation of human T cells with superantigen SEA. Methods　T cells proliferation, IL-2 production, DNA synthesis, cell phenotype and apoptosis induced by the SEA in the first or restimulation were detected. Results　 It was shown that 100 ng/mL was the optimal activation concentration, IL-2 production arrived at top level in the third day, SEA activated CD4+ and CD8+T with the same degree, T cells start apoptosis in response to SEA restimulation within 24 hours and apoptosis disappeared through addition of rhIL-2. Conclusions　 There were correlation between activation and SEA concentration or stimulation period； SEA activated both CD4+ and CD8+T cells without change the cell phenotype.

Objective To evaluate the effects of flurbiprofen axetil and fentanyl in postoperative analgesia on immune function and stress response of the patients undergoing esophagectomy. Methods Sixty patients were randomly divided into three groups with 20 cases in each group, including Group F_1 (pre-operative: flurbiprofen axetil 50mg, postoperative : flurbiprofen axetil 50mg + fentanyl 10μg/kg + droperidol 2.5 mg), F_2 (postoperative: flurbiprofen axetil 100mg + fentanyl 10μg/kg + droperidol 2. 5 mg) , and group C (postoperative: fentanyl 10μg/kg + droperidol 2.5 mg). The VAS score was recorded at 1, 24, 48 hours after surgery. Blood samples were obtained from peripheral vein for determination of NE, ACTH, COS, CD3~ + , CD4~+ , CD8~+ and CD4 VCD8~+ at 30min before surgery, 1 d, 2d after surgery. Results Patients in the three groups did not show any significant difference in the VAS scores ( P > 0.05). NE was significantly lower in group F_1 than group F_2 and group C at 1 d after surgery ( P < 0. 05). There were significantly decreased ACTH in group F_2 and F_1 than group C at 1d after surgery( P <0. 05), and it was significantly decreased in group F, than that in group C at 2d after surgery( P < 0.05). COS was significantly decreased in group F_1 than that group C at 1d after surgery( P <0.05 ). CD3~+ T-lymphocytes were significantly higher in group F_2 and F_1 than that group C at 1h after surgery ( P <0. 05) , and group F, was significantly higher than group C at 2d after surgery( P <0.05). CD4~+ T-lymphocytes were significantly increased in group F_1 than that in group C and F_2 at 1d after surgery( P < 0.05). CD8~+ T-lymphocytes were no significantly change in 3 groups and at each time point ( P >0.05). CD4~+/ CD8~+ were significantly higher in group F_1 than that in group C and F_2 at 1 d after surgery( P <0.05). Conclusion Postoperative analgesia by using flurbiprofen axetil and fentanyl can diminish the using dose of postoperative opoiod drug, it can decrease patients postoperative stress level and improve patients cellular immune function.

Objective:To establish a patient-derived xenotransplantation (PDX) animal model of gastric cancer, and observe the anti-cancer effect of chemotherapeutic drugs on this model.Methods:Human gastric cancer tissues were inoculated into the subcutaneous tissues of both axillaries of NOG mice and were subcultured for 3 generations. The tumor tissues of the third-generation NOG mice were selected and inoculated into the subcutaneous tissues of left axillary of 21 severe combined immunodeficiency-non-obese diabetes mellitus (SCID-NOD) mice to establish PDX mouse model of gastric cancer. The inoculated mice were divided into control group (mice received only 0.9% sodium chloride injection), oxaliplatin group, cisplatin group, paclitaxel group, fluorouracil group, tegafur, gimeracil and oteracil porassium capsules group and capecitabine group, with 3 mice in each group, and the corresponding drugs were given. The mice survival status, tumor volume and tumor weight at different times were recorded. Mice were sacrificed on the 61st day of administration, and the tumor inhibition effects of 6 kinds of chemotherapy drugs on the PDX model of gastric cancer were evaluated.Results:After being subcultured for 3 generations, the stability of tumor transmission in PDX animal model of gastric cancer was improved, and the homogeneity of tumor growth was good at the initial stage. At the early stage of administration, the model was more sensitive to oxaliplatin, fluorouracil and capecitabine, and the tumor growth inhibition (TGI) values on the 31st day were 63.37%, 52.11% and 78.48%, at the end of administration, the model had the best sensitivity to capecitabine with a TGI value of 59.22% and a tumor inhibition rate of 58.65% on the 61st day. The TGI curve after administration showed that paclitaxel had no obvious anti-tumor effect, cisplatin had the worst anti-tumor effect, and the model had poor tolerance to tegafur, gimeracil and oteracil porassium capsules.Conclusion:The PDX animal model of gastric cancer is successfully established, and capecitabine has the best tumor suppressive effect on this model.

Purpose To investigate the expression of Apaf-1 and Caspase-9 in prostate cancer ( PCa) and benign prostatic hyperplasia ( BPH) , and to analyze their correlation with clinicopathological parameters of PCa. Methods Immunohistochemistry was used to de-tect Apaf-1 and Caspase-9 protein in 45 cases of PCa and 60 cases of BPH. Results The positive rate of Apaf-1 and Caspase-9 in PCa tissues was significantly lower than that in BPH (P<0. 05). The expression of Apaf-1 and Caspase-9 was not correlated with the age of patients and distant metastasis (P>0. 05), but it was correlated with the pathological grade and clinical stage of PCa (P<0. 05). Conclusion Apaf-1 and Caspase-9 are lowly expressed in PCa. There is positive correlation between the expression of Apaf-1 and Caspase-9 (rs =0. 645, P<0. 01). Apaf-1 and Caspase-9 might be correlated with the carcinogenesis and development of PCa.

Objective To establish a stable animal model of transplanted human colorectal cancer.Methods BALB/c nude mice were randomly divided into orthotopic colorectal model group and subcutaneous inoculation model group by random number table,and separately inoculated with 0.1 ml human colon cancer cell HCT116 under the density of 2×107/ml into the orthotopic colorectal (with the self-made inoculator) and right forelimb pit subcutaneously.The mice were observed for 60 days to compare the tumor formation and tumor growth in the two groups.Results The tumor formation rate of all 18 animals was 100 ％ (18/18) in the orthotopic colorectal group.The average tumor weight was (2.78±1.86) g and the average survival time was (45.00±11.99) d.The tumor formation rate was 27.78 ％ (5/18) in the subcutaneous inoculation group.The average tumor weight was (1.74±0.82) g,and the average survival time was (60.00±0.00) d.Conclusion 0.1 ml (2×107/ml) human colon cancer cell suspension HCT116 inoculated into BALB/c nude mice orthotopic colorectal with self-made inoculator could establish human colorectal cancer animal model successfully.

Objective To observe the effects of brazilin on proliferation and apoptosis in T24 cells.Methods Trypan blue exclusion test was performed to detect the inhibition of brazilin on the growth of T24 cell lines in vitro cultured within different time.After exposure to different concentrations of brazilin,homogeneous bioluminescence assay was used to detect the inhibitory action of brazilin,Caspase-3 and Caspase-9 activity on T24 cells.Cellular apoptosis was measured by flow cytometry (FCM) and observed by laser scanning confocal microscope.Results Brazilin could significantly inhibit the proliferation of T24 cells after 8 hours,the inhibitory rates of the brazilin at concentration of 25,50,100,200 μg/ml against T24 cells respectively were 43.19 ％,60.73 ％,86.38 ％ and 93.89 ％ (P ＜ 0.05).After exposured to 50 μg/ml of brazilin,the inhibition ration to T24 cells increased with time prolonging (52.72 ％ in 4 h,60.73 ％ in 8 h,91.77 ％ in 24 h,96.41％ in 48 h) (P ＜ 0.05).The activity of Caspase-3 and Caspase-9 increased slightly when brazilin was at 25 μg/ml,but there was no statistical differences compared with that in the control group (P ＞ 0.05).When cells were treated with an increase of the concentration of brazilin from range of 7.5-60 μg/ml for 16 hours,the apoptosis ratio in turn showed a upward trend of 0.15 ％,1.35 ％,2.91％,34.76 ％.It could be seen by laser scanning confocal microscope that the apoptosis occurred in the cells.Conclusion Brazilin can effectively inhibit the proliferation of T24 cells and induce apoptosis in a dose and time dependent manner.

Objective:To explore the relationship between transcription factor AP 1 and NF kappaB and IL 2 decrease in anergic T cells.Methods:Nuclear proteins of activated T cells and anergic T cells were extracted and then the expression of AP 1 and NF kappaB were determinated by gel electrophorotic mobility shift assay.Results:Compared to activated T cells,anergic T cells expressed defective AP 1 transcription factor;there were three forms NF kappaB complex in both activated T cells and agergic T cells,but NF kappaB transcription activity was higher in anergic T cells than that in activated T cells.Conclusion:It has been demonstrated that these alteration of transcription factors have been likely to be responsible for repression of IL 2 in anergic T cells.

Objective To construct staphylococcal enterotoxin D (SED) mutants expressed in Escherichia coli ( E. coli ). Methods The expected mutants were introduced into the SED DNA by megaprimer PCR method. The PCR products ligated to plasmid pTrcHis B were transformed into E. coli DH5? for IPTG induced expression. The target protein was purified by Ni NTA metal affinity chromatography and analyzed by SDS PAGE and immunoblotting. Results The sequencing results showed that mutant nucleic acids were successfully introduced at the expected sites of SED gene. SDS PAGE and immunoblotting confirmed that the proteins of SED mutants were obtained by Ni NTA metal affinity chromatography. The mutants were named as SEDN23A, SEDN23A/H26R, SEDF45A, SEDL59A, SEDN61A, SEDI92A, and SEDF203A, respectively. Conclusion Several SED mutants are successfully constructed, which lays a foundation for subsequent studies of immune recognition of SED.

Objective To establish a method for preparing orthotropic transplantable hepatocellular carcinoma in mice. Methods According to the liver detailed anatomical structure of the mouse, 50 μl mouse ascites containing 1 ×106 and 5 ×105 mouse hematoma H22 cells was input to liver in 12 Kunming mice through percutaneous intraperitoneal injection by syringe, respectively, to establish orthotopic transplantable hepatocellular carcinoma model. The growth status of mice was observed, and the pathological changes of liver and tumor metastasis tissues were detected. The tumor formation and metastasis were analyzed. Results The tumor formation rate was 100% (12/12) by direct injection of mouse hematoma H22 cells in 2 groups. The inoculated mice started to appear ascites at the 6th day, and all mice produced ascites at the 10th day. The survival time was (16.17 ±3.07) d and (18.08 ±3.34) d in 1 ×106 group and 5 ×105 group, respectively. Some mice emerged tumor metastasis in kidney, intestine, spleen and mesenteric lymph nodes. Conclusion The method of direct injection could establish orthotropic transplantable hepatocellular carcinoma model in mice, which can be used for antitumor drug research.