Objective:To study the expression and correlation of tumor-associated macrophages(TAM) and CCL5 in ganstric cancer.Methods:48 cases patients with completed clinical and pathological data of gastric cancer paraffin block specimens were select-ed.Cancer tissues and adjacent tissues were used as control,using SP immunohistochemical method to detect CD68 and CCL5 in gastric cancer tissues and adjacent tissues,and using the Spearman correlation statistics statistical methods for the correlation.Results:CD68 and CCL5 showed positive expression in gastric cancer tissue,significantly higher than those in the adjacent tissues(P<0.01),CD68 and CCL5 were related with gastric cancer invasion depth, lymph node metastasis, TNM stage and tumor differentiation ( P<0.001 ) . There was positive relation between the expression of CD68 and CCL5 in gastric cancer(P<0.01,r=0.759).Conclusion: CD68 and CCL5 played a driving role to the invasion and metastasis of gastric cancer occurrence,suggesting that the secretion CCL5 by TAM may promote the invasion and metastasis of gastric cancer.

Objective To analyze the clinical features of eosinophilic granulomatosis with polyangiitis (EGPA).Methods The clinical characteristics,laboratory test,therapeutic regimen,pathology and imaging diagnosis in 23 cases with EGPA from the First Affiliated Changhai Hospital of Second Military Medical University within June 2007 to January 2013 were retrospectively analyzed.Results There were 14 males and 9 females,with the age ranged from 21 to 68 years.The mean age was (42±9) years.The overall prevalence of asthma was 78％(18/23).The allergic rhinitis accounted for 61％(14/23).The maxillary sinuses were the most frequendy involved,which accounted for 57％(13/23).Skin involvement was 57％(13/23),peripheral neuritis was 70％(16/23).Central nervous system involvement presented cerebrovascular event.Cardiac involvement accounted for 48％(11/23),digestive system involvement accounted for 17％(4/23).The outcome of auxiliary laboratory test revealed that 96％(22/23) patients expressed significantly higher levels of IgE and 70％(16/23) patients carried anti-neutrophil cytoplasmic antibodies (ANCA) which were presented as the perinuclear ANCA (p-ANCA).The patchy infiltrates of lung CT scan accounted for 69％ (11/23).EMG showed mononeuritis multiplex and symmetric sensory motor neuropathy.The abnormal ratios of ECG and Echocardiography were 48％(11/23),79％(11/14) respectively.The pathological manifestations were necrotizing vasculitis,eosinophilic tissue infiltration,and extravascular granulomas.Conclusion Our results confirm the heterogenicity in clinical presentation and lack of specificity.Early diagnosis and treatment will be helpful for good prognosis.

Objective: To investigate the effect of cochinchina momordica seed ethanol extract (CMSEE) on the proliferation of melanoma B16 cells and the underlying mechanism. Methods: MTT and clone formation assay were used to assess the effect of CMSEE on the growth of B16 cells. Morphological changes of B16 cells were observed under phase-contrast microscope and Giemsa staining. Cell cycle and apoptosis rate were examined by flow cytometry (FCM). The effect of CMSEE on melanin production and tyrosinase activity of B16 cells was assessed by colorimetry. The effect of CMSEE on the expression of C-myc, P38, and Tyr genes was examined by RT-PCR. Results: CMSEE (10-100 mg/L) inhibited the proliferation of B16 cell in a dose-and time-dependent manner. After treatment with 10-40 mg/L CMSEE, B16 cells showed typical differentiation morphology, and melanin production and tyrosinase activity were increased. B16 cells treated with 100 mg/L CMSEE showed apoptotic morphology, decreased melanin production and tyrosinase activity. B16 cell number in G_0/G_1 phase was significantly increased (P<0.01); C-myc mRNA expression was down-regulated, and P38, Try mRNA expression was up-regulated in B16 cells after treatment with 10-40 mg/L CMSEE. Conclusion: CMSEE can markedly inhibit the proliferation of melanoma B16 cells, which is related to induction of differentiation and promotion of apoptosis of B16 cells.

Objective:To study the inhibitory effect of IL-27 against human tumor angiogenesis and the related mechanisms.Methods: Human esophageal carcinoma cells(Eca109/IL-27) stably transfected with IL-27 gene were injected into nude mice to establish tumor-bearing mouse model.The survival time and tumor growth were observed.IFN-? level secreted by splenocytes was measured by ELISA.Expression of VEGF and CD34 was detected by immunohistochemistry method and MVD was calculated according to CD34 level.RT-PCR was used to detect the expression of IP-10 and MIG mRNA in the tumor tissues.Results: The survival time of mice injected with Eca109/IL-27 cells was significantly longer than those of mice injected with wide type Eca109 or Eca109/LXSN(blank vector) cells(P

Objective:To observe the effect of FSEER on the immunosuppressive and bone-marrow-suppressive mice after chemotherapy,and explore the mechanism of hematopoietic and immunologic function in mice accentuated by FSEER.Methods: Mice were injected cyclophosphamide(Cy)except control group,then randomly divided into mode group(saline),FSEER groups[120,60 mg/( kg· d) ].The spleen index( SI) of all mice was calculated respectively.Flow cytometry instrument testing mice peripheral blood lymphocytes CD3,CD4,CD8 change.The content of TNF-α,IL-2 and IL-12 in mice serum were measured by ELISA kits.Morphological images of bone marrow of the mice were observed under the microscope after Wright-Giemsa′s staining.Results:The spleen index( SI) was increased in both of the two FSEER groups.ELISA analyses showed that the content of TNF-αand IL-2 was increased in both of the two FSEER groups.The population of CD3+CD4+T lymphocyte and the ratio of CD4+/CD8+were all increased in the low dose experimental group.After treated with FSEER, the hematopoietic depression was improved significantly.Conclusion: FSEER can improve the state of immunosuppressive and myelosuppressive mice caused by Cy thus could alleviate the side effect of chemotherapy ef-fectively.

Objective:This work aims to detect the levels of miR-200c/141 methylation and miR-200c/141 in gastric cancer tissue and investigate the relationship between miR-200c/141 expression and clinical parameters. Methods:The methylation status of miR-200c/141 CpG island and miR-200c/141 in gastric cancer tissue specimens was evaluated by qRT-PCR or BS-MSP method. We analyzed the relationship among the methylation status of miR-200c/141 CpG island, expression level of miR-200c or miR-141, and clinical parame-ters. Results:The status of miR-200c/141 CpG island methylation in gastric cancer tissue was significantly higher compared with that in paracarcinoma tissue. MiR-200c and miR-141 were markedly decreased in gastric cancer tissue compared with those in adjacent tis-sue. MiR-200c/141 CpG island methylation was negatively related with the expression of miR-200c and miR-141 in gastric cancer speci-mens. Conclusion:The upregulation of miR-200c/141 CpG methylation inhibits miR-200c/141 expression in gastric cancer tissue.

Objective:Effect of ICA (Icariin) was observed on immunosuppression mice caused by Cyclophosphamide(Cy).Methods:A total of 60 mice,expect control group mice,were injected Cy intraperitoneally [300 mg/(kg·d)],then randomly divided into modelgroup(saline),positive (shenqi,1 ml/d)groups,ICA groups[150,80,40 mg/(kg·d)].Saline,shenqi and ICA were perfused respectively for 10 successive days;All mice were sacrificed after 12 h of last administration for performing following experiments.Measuring SI and TI and calculating amounts of lymphocyte from spleen and peripheral blood;Proliferative reaction of splenic lymphocyte stimulated by ConA and LPS were detected by MTT assays.Killing activity of NK and CTL cells for colon26 and level of TNF-α production were measured by LDH method.Ratios of CD3~+ T and NKT in splenic lymphocyte (SPL) were studied by FACS.Results:ICA can recovery TI and SI of mice to normal level and boost the population of splenic lymphocyte (SPL) (P<0.01).After treatment with ICA,activities of NK cell and CTL cell were significantly improved,and the level of TNF-α was significantly elevated.The ratios of NKT and CD3+T in the ICA-treated group were increased obviously (P<0.01).Conclusion:ICA can enhance immunological function and alleviate the side effect of chemotherapy effectively.

Objective To investigate the effects of 2-methoxyestradiol (2-ME) on the proliferation and apoptosis of a mouse malignant melanoma cell line B16,and to explore their mechanism.Methods B16 cells were cultured in vitro,and divided into a negative control group receiving no treatment and several intervention groups treated with 2-ME at final concentrations of 5,10,20,40 mmol/L,respectively.After different durations of treatment,inverted phase-contrast microscopy was conducted to observe the morphologic change of B16 cells,sulforhodamine B (SRB) assay to evaluate proliferative activity and to draw growth curve of B16 cells according to the absorbance value at 490 nm,flow cytometry to detect cell cycle and apoptosis,and reverse transcription PCR and real-time PCR were performed to measure the expressions of the apoptosis-inducing gene gadd45b and proto-oncogene c-myc.Results As repeated measures analysis of variance showed,there were significant differences in the inhibitory effect on B16 cell proliferation among different concentrations (5,10,20,40 mmol/L) and different treatment durations (24,48,72 hours) of 2-ME (F =1170.94,1843.04,respectively,both P ＜ 0.01),and there was a significant interaction effect between these concentrations and treatment durations (F =272.79,P ＜ 0.01).After 48-hour treatment with 2-ME at 10,20 and 40 mmol/L,the apoptosis rate of B16 cells was increased to (4.13 ± 1.12)％,(11.25 ± 2.380)％ and (19.46 ± 2.9)％ respectively,compared to (0.23 ± 0.5)％ in the negative control group (all P＜ 0.01); the proportion of B16 cells in G0/G1 phase was increased to (59.5 ± 5.6)％,(63.4 ± 8.2)％ and (70.8 ± 4.4)％ respectively,compared to (44.1 ± 3.4)％ in the negative control group.There was a significant difference in the proportion of B16 cells in G0/G1 phase among the negative control group and intervention groups (F =13.56,P ＜ 0.05).Moreover,the mRNA expression of gadd45b was significantly enhanced after 24-hour treatment with 2-ME at concentrations of 20 and 40 mmol/L (both P＜ 0.01),while that of c-myc was significantly weakened after treatment with 2-ME at 10,20 and 40 mmol/L (all ＜ 0.05) compared with the negative control group.Conclusion 2-ME can inhibit the proliferation of B16 cells in vitro,upregulate the expression of gadd45b gene and downregulate the expression of C-myc gene.

Objective:To compare and observe the different clearance effect of milk containing anti-Helicobacter pylori specific antibody. Methods:Four H. pylori strains were used to immune dairy cows to obtain milk containing anti-Helicobacter pylori specific antibody,of which,one was standardized strain and the other three were locally epidemic. Totally 148 people were screended,in which 72 were C-14 urea breath test positive, finally 39 meet the criteria. They were divided into two groups, the test group contained 21 subjects,were treated milk containing anti-Helicobacter pylori specific antibody;the 18 subjects in control group with common milk. The study was continued for 2 months. Results:Conducting the C-14 urea breath test,9 subjects in test group were negative,but no one was changed in control group. The effective clearance rate of the test group was 42. 86%,and there was no effective clearance in the control group,so there was significant difference in the two groups(P=0. 005,P<0. 05). Conclusion: The milk containing anti-Helicobacter pylori specific antibody is polyclonal and has higher valence,and could clear H . Pylori effectively.

ObjectiveTo evaluate the adverse events occurred during tumour necrosis factor (TNF)-αblocker treatment in Chinese Han population patients with ankylosing spondylitis (AS).MethodsThis study had enrolled 369 Chinese Han population patients with ankylosing spondylitis.They all received TNF-αblocker treatment in the hospital.All 1011 administration were recorded in total.All of them were evaluated for adverse events 2 hours after injection,126 of them had received long-term TNF-α blocker injection,and they were followed-up at week 8,12,52,104.Mild immediate adverse events and long-term adverse events were all counted.SPSS 10.0 software package was used for Fisher's exact test.ResultsThree hundred and sixty-nine patients had 1011 administrations in total,652 had received rhTNFR:Fc,316 had infliximab,21had etanercept,22 had adalimumab injections.Adverse events 2 hours after injection were:17 (2.6％) for rhTNFR:Fc,12 (3.8％) for infliximab,0 for etanercept,1 (4.5％) for adalimumab.Twenty adverse events were mild(12 for rhTNFR:Fc,9 for infliximab),5 events were moderate(3 for rhTNFR:Fc,1 for infliximab,1 for adalimumab),4 events were severe(2 for rhTNFR:Fc,2 for infliximab).The frequency of adverse events were comparable between rhTNFR:Fc and Infliximab injection in immediate adverse reactions (P=0.31).One hundred and twenty-six (69 rhTNFR:Fc,57 infliximab) patients had long-term usage,and were followed-up at week 8,12,52,104,39 patients had adverse reactions:20 (51.3％) for rhTNFR:Fc,19(48.7％) for infliximab.Thirty-seven patients had infectious events(94.9％ ),1 neurological event(2.6％),and 1 patient had tuberculosis relapse (2.6％).Outcomes were comparable with rhTNFR:Fc and infliximab in long-term usage(P=0.69).ConclusionAttention should be paid to the above events in Chinese Han patients with ankylosing spondylitis who were treated with TNF-α blocker treatment.Special attention should be paid to those patients who are in their third or fourth injection.The occurrence of immediate reaction or long-term adverse events between rhTNFR:Fc and infliximab are comparable.